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ORGANIZING COMMITTEE

Chief Patron H.H. Swami Poornamritananda Puri Patrons Dr. Venkat Rangan, Vice Chancellor, Amrita Vishwa Vidyapeetham Dr. Prem Nair, Medical Director, Amrita Institute of Medical Sciences Dr. Prathapan Nair, Principal, Amrita School of Medicine Organizing Secretary Prof. D. M. Vasudevan Joint Organizing Secretary Dr. Subir Kumar Das Dr. Kannan Vaidyanathan Executive Committee Members Dr. Subhakumari K.N (Chairperson) Dr. T.Vijayakumar Dr. Subramanya Iyer Dr. R.Krishna Prasad Dr. Deepthi Menon Chairpersons of Committees Reception, Hospitality and Food: Dr. Sajitha Krishnan, Dr. P. Sujaya, Dr. Leela Menon Exhibition : Dr. Indira Samal Corporate Liaison:Dr. Ramesh Kumar, Mr. M.V Thampi Accommodation and Transport: Dr. (Mrs) P.G Nair

Scientific Committee : Dr. S. Sreekumari CME : Dr. V. Balakrishnan Workshop : Dr. Krishnakumar Menon Treasurer : Dr. P.N.K Menon Cultural : Dr. Jagath Lal

ACBICON 2009 LOCAL ADVISORY BOARD Dr. Asuma Beevi, Dr. Beena.C, Dr. Geeta.A, Dr. Geetha.A, Dr. Geetha Damodaran, Dr. George Abraham, Dr. Gopinathan Nair.S, Dr Jaya.S, Dr. Jayanthi Bai, Dr. Jeeji Palokaren, Dr. Jose Jacob, Dr. Jose Raj, Dr. Minnie Mary Mammen, Dr. Miriam Varkey, Dr. Mohanan, Dr. Parvathy, Dr. D. Prasanna, Dr. C.S Poulose, Dr. Pushpalatha, Dr. Rajasekharan Pillai, Dr. K.Ramachandran, Dr. Sajeevan, Dr. Sankar Saidharan, Dr. Sathyasudha, Dr. N.Sreedevi, Dr. S.Sreekumari, Dr. Sreekumar, Dr. P.R Sudhakaran, Dr. B.Sulekha, Dr. Sundara Devi, Dr. T.Vijayakumar, Dr. Vijayalekshmi ACBICON 2009 NATIONAL ADVISORY BOARD Dr T. Ashavaid, Prof. S.P Dandekar, Dr. T.P.A Devasagayam, Prof. Dinesh Puri, Dr U.N Donde, Prof B.C Harinath, Prof. A.S Kanagasabapathy, Prof. Krishnajyoti Goswami, Prof. M.V. Kodliwadmath, Prof. P.C Mahapatra, Prof. Abbas Ali Mahdi, Prof. T. Malati, Prof. K.L Mukherjee, Prof. P.S Murthy, Prof. N.C Panda, Prof. T.N. Pattabhiraman, Prof. Praveen Sharma, Prof. D.N. Rao, Prof. Sandip Bandopadhyay, Prof. U. Satyanarayana, Dr. R. Selvakumar, Prof. P.P. Singh, Prof. K.P. Sinha, Brig. (Dr) Y.V. Sharma, Prof. P.S. Shukla, Prof. K.K. Srivastava, Prof. L.N. Srivastava, Prof. Subrata Chattopadhyay, Prof. Subrata Sinha, Prof. B.N. Tiwari, Prof. Uday Kumar, Prof. P. Usha Sarma, Prof. T. Venkatesh, Prof. H.S. Virupaksha. All founder members, All Past presidents and members of various committees of national body. ACBICON 2009 INTERNATIONAL ADVISORY BOARD Prof. Amitava Dasgupta, Dr. Beremans Lopez, Dr. Elizabeth Frank, Dr S Jamal Raja, Prof. Liaquat Ali, Dr. Lindsay Brown, Dr. Michel Silvestri Joseph, Prof. Dr. Md. Suhrab Ali, Dr V Parameswaran, Dr. Perry Gotlesfeld, Dr. Samuel Vasikaran, Dr. Tom Hartley, Dr Udayan Ray

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ORATIONS

O-1. : Dr. T.N Pattabhiraman Oration Laboratory Overload: How to Optimize? Brig. M.M. Arora O-2. : K.E.M. Hospital and Seth G.S. Medical College Oration Regenerative Nanomedicine - Prospects for Diagnostics and Therapy using Nanomaterials Shanti Kumar Nair O-3.: Mrs. & Dr. G.P Talwar Oration Endothelial Dysfunction with Special Reference to eNOS Gene Polymorphism: Another Plausible Mechanism for CAD Risk in Postmenopausal Women Jayashree Bhattacharjee O-4. : K.L Gupta Memorial Oration Tyrosine Kinases from Basic Science to Therapy T.S.Ganesan O-5- :Awadesh Saran Memorial Oration Lead Poisoning: Current Status in Developing Countries and the Global Perspective. Thuppil Venkatesh

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KEY NOTE ADDRESSES

KN-1. Vitamin E is activated by phosphorylation to alphatocopheryl phosphate Angelo Azzi KN-2. Impact of Pharmacogenetics on Therapeutic Drug Monitoring M. Oellerich, V.W. Armstrong and N. von Ahsen

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INVITED LECTURES

IL-1.:The role of Bone Turnover Markers in Clinical Management of Osteoporosis SD Vasikaran IL-2. : A New Diagnostic Paradigm: Some Enabling Technologies Ross Barnard IL-3: The Natural Phenolic, Malabaricone B Selectively Kills the Cancer Cells through Mitochondrial Pathway S. Chattopadhyay

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IL-4: Marine Brown Algae with Potent Antioxidant and Radical Scavenging Activities M.S.Kanthimathi, Ghaitri Nadarajan and Khoo Kong Soo IL-5: Male Infertility: Role of Oxidative Stress and Some Possible Herbal Remedies Abbas Ali Mahdi, Mohd. Kaleem Ahmad, Kamla Kant Shukla, Singh Rajender, Satya Narain Shakhwar and Sohail Ahmad*** IL-6: [(E)-HST-1], 3' -5' Hydroxylated Congener of Resveratrol, A Better Anticancer Agent Sandip K Bandyopadhyay IL-7: Oxidative Stress Management: Therapeutic Approach for Depression Naheed Banu, Ayesha Zafir, Shoa Naqvi and Ghazala Mehdi IL-8: Antioxidant Potential and LDL Oxidation in Disease. Dr. Neelima Singh and Nivedita Singh. IL-9: Total Quality Management in Health Laboratories Dr. Ghassan Shannan IL-10: Beyond Means and SDS: Recommended Statistics for Knowledge Extraction from Accumulated Pathology Data and Your Practice Experience T F Hartley IL-11: Web Based Education and Training in Clinical Laboratory Services Dr. Uday Donde IL-12: Lean - Sigma Metrics Protocol of Total Quality Management in a New Laboratory Set-up Barnali Das* IL-13: Laboratory Management of the Metabolic Syndrome Sridevi Devaraj IL-14: Thyrotropin Secreting Pituitary Adenomas (TSHoma) V.Parameswaran, G.Stilwell and J.Burgess IL-15: Monogenic Disease in Endocrinology Tjin-Shing Jap IL-16: Obesity ­ A Global Epidemic Dr.Nalini Ganesan IL-17: Novel Interactive Signal Molecules in the Hypothalamus Regulating GNRH Release Dr. Edathil Vijayan

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IL-18: Role of Ethanol in Inducing Inflammatory Brain Damage and Vasculogenesis Sukhes Mukherjee, Subir Kumar Das, Subrata Chattopadhyay and D.M Vasudevan IL-19: Culture of Rat Gastric Mucosal Cell Line RGM1 or its Tumor Cell Line RGK1 Combined with Human Fibroblasts Hideyuki J Majima, Yoko Yamaguchi, Mitsuhiro Ohshima, Hiroko P Indo, Kazuo Tomita, Yoichiro Iwashita, Hsiu-Chuan Yen, Takuro Kodama, Masakazu Souda, Akihide Tanimoto, Motoharu Miymoto, Kazuyuki Noguchi, Osamu Shimokawa Yumiko Nagano, Tsuyoshi Kaneko, Hirofumi Matsui, and Toshihiko Ozawa IL-20: Cancer Biomarkers of Prognostic Value Paturu Kondaiah IL-21: HPV 16 Variants in Normal, SIL and Cervical Cancer Women. Neeta Singh, Chandresh Sharma, Archna Singh Palika Datta and Sunesh K. Jain IL-22: An Update in Molecular Oncology: Cervical Cancer B.Nagarajan IL-23: Natural Products as Potential Treatments for the Human Metabolic Syndrome Lindsay Brown IL-24: Association of Cytokines and Insulin Resistance in NonAlcoholic Fatty Liver Disease Subir Kumar Das IL-25: The Promise of Stem Cells and Gene Therapy as Novel Approaches to the Treatment of Multiple Sclerosis (MS) Natalie Payne, Christopher Siatskas, Guizhi Sun, Daniella Herszfeld, Natalie Seach, Sara Litwak, Martin Short, Ban-Hock Toh, Frank Alderuccio, Thomas Backstrom, Richard L Boyd, and Claude C A Bernard IL-26: Hs-CRP, Inflammation and Its Role in Coronary Heart Disease Arun Raizada, H V Singh and Naresh Trehan IL-27: Pro-oxidant Activity of Artemisinin Causes an Apoptosis Like Death in Leishmania parasites Mitali Chatterjee and Rupashree Sen IL-28: Novel Filarial Genes or Recombinant Filarial Proteins as Effective Vaccine Candidates MVR Reddy

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IL-29: Bilirubin from Toxic Metabolite to an Antioxidant: Mechanisms S. Adhikari IL-30: New Perspectives on the Clinical Importance and Applications of Vitamin D Testing H.A Morri IL-31: An Insight to the Mechanism of Lycopene: A Biocarotenoids Against ã-Radiation Induced Apoptosis; MAPK Signaling Pathways and DNA Repair Enzymes in Normal Lymphocytes and Lymphoblastoid TK-6 Cell Lines in Vitro. Venugopal P. Menon IL-32:The Role of CRP in Atherothrombosis Ishwarlal Jialal IL-33: The Clinically Effective Use of Available Cardiac Biomarkers. Ray U IL-34: Mycobacterial ES-31 Serine Protease of Interest in Diagnosis of Tuberculosis and HIV-TB Co-infection and with Potential as Drug Target B.C. Harinath IL-35: Autoantibodies: Is it Possible to Predict Autoimmune Diseases? Marcos López-Hoyos IL-36: Cell-Cycle Synchronization in Antinuclear Antibodies Detection Petraki Munujos Vinyoles IL-37: Importance of Monitoring Magnesium Levels in Cardiac Conditions Dr. S. P. Dandekar IL-38: Tissue Zinc Levels in Carcinoma Dr. R. Selvakumar IL-39: Prenatal Screening for Chromosomal Disorders Päivi Laitinen IL-40: Routine Screening of Newborns for Metabolic Disorders using Tandem Mass Spectrometry Rohit Cariappa IL-41: Diagnosis of Aminoacidurias by HPLC: A Few Interesting Cases Dr. Kannan Vaidyanathan

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IL-42: Specific Plant-Derived Products with Relevance to Clinical Medicine PG Latha IL-43: Natural Products and Therapeutic Strategies: A Mechanistic Approach Bipin Nair IL-44: ABC of Monoclonal Gammapathy and its Indian Scenario T.Malati IL-45: The Clinical Significance of Microalbuminuria T.Malati IL-46: Biochemical Changes in Mountain Sickness Savitha Nagabhushan, Thuppil Venkatesh, Vidyasagar Casikar E Mujica , J. Aliaga , N.Lopez , C.Smith , F. Bartholomew and Gustavo Zubieta-Calleja (Jr), Gustavo Zubieta-Castillo (Sr), and Poul-Erik Paulev M.Mongelli IL-47: The High Altitude Adaptation Formula GR Zubieta-Calleja, P.E. Paulev, L Zubieta-Calleja & G.ZubietaCastillo IL-48: Do Over 200 Million Healthy Altitude Residents Really Suffer from Chronic Acid- Base Disorders? Gustavo Zubieta-Calleja (Jr), Gustavo Zubieta-Castillo (Sr), and Poul-Erik Paulev, High Altitude Pathology Institute, La Paz, Bolivia. [email protected] IL-49: The Ventilatory Hyperoxic Shunt Test at 3510 m. of altitudeG Zubieta-Castillo, L. Zubieta-Calleja, and GR ZubietaCalleja, IL-50: Get Together on Biochemistry - An Experimental Teaching Dr.W.Ebenezer William IL-51: The Role of Clinical Chemist in the Management of PatientRay U IL-52: Maturity Onset Diabetes in the Young (MODY) V.Parameswaran, A.Duffield, S.Powell, J.Burgess IL-53: Post -transplant Diabetes Mellitus Following Immunosuppressive Therapy Dr. Pragna Rao IL-54: Point-of- Care testing (POCT) C.V.Anand

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IL-55: Protein Misfolding Diseases Dr.Usha Anand IL-56: Obesity ­ A Global Dilemma V.Parameswaran, L.Blackwell, L.Diggle, J.Burgess and R.McCallum IL-57: Scholarly Communication: Writing a Research Paper for a Peer-Reviewed Publication R S Jayasomu IL-58: Urine Albumin: Standardisation and Reporting Issues. H. D. Martin IL-59: Molecular Mechanisms of Angiogenesis in Heart and Lung Hari S. Sharma IL-60: The Pro-Inflammatory Role of Extracellular RNA in Arteriogenesis Elisabeth Deindl IL-61: Pharmacogenomics: One Size Does Not Fit All Dr. Amit Chakrabarti. IL-62: Lead Not Safe in Environment Dr. Krishnajyoti Goswami IL-63: Developments in the Biochemical Assessment of Renal Disease David Torrens IL-64: ISO 15189 : 2007 Dr. Elizabeth A. Frank ABSTRACTS OF PRESENTATIONS Abstract No: 1.1: A Study on Free and Total Prostate Specific Antigen in Benign Prostatic Hyperplasia (BPH) and Carcinoma Prostate Dr.R. Lalitha Abstract No: 1.2: Evaluation of Enzyme Markers in Carcinoma Cervix. Dr.P.B.Desai and Dr.Chetana.K Abstract No: 1.3: Serum Fluoride and Sialic Acid Levels in Osteosarcoma R. Sandhu, Z. S. Kundu, S. Kharb and Harbans Lal Abstract No: 1.4: Diagnosis of Breast Cancer by the Determination of Plasma Maspin Level Karabi Ganguly (Bhattacharjee), G.V.Girish and Asru K. Sinha

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Abstract No: 1.5: Evaluation of Serum Tumor Markers and Fertility Hormones in Postmenopausal Women with Ovarian Cancer L. Vishnu Prasad and Dr.R.Arivazagan Abstract No: 1.6: Serum sialic acid forms as tumors head, neck, thyroid & lung tumors. Mahendra R. Pakhale , Dr.Murhar and Rajesh Rajrkar Abstract No: 1.7: Matrix Metalloproteinases and Their Inhibitors: Correlation with Invasion and Metastasis in Oral Squamous Cell Carcinoma Ragini D. Singh, N Haridas, Shilin N. Shukla, Jayendra B.Patel, Bhairavi N. Vajaria and Prabhudas S. Patel Abstract No: 1.8: A Study of Oxidative Stress in Cancer Patient before and after Supplementation of Vitamin C and E Dr. Pranami Bordoloi, Dr. Bikash Kr. Chaudhury, Dr. Chandan Kr. Nath and Dr. Dipali Das Abstract No: 1.9: Role of Serum Enzymes in Oral Cancer. Dr P.B Desai and Dr Shikha Sharma Abstract No: 1.10: Biochemical and Hematological Markers in Patients with Chromosome 22q11.2 Deletion and Conotruncal Defects. Alka Anilkumar, M.V.Thampi, R.Krishna Kumar, K.Mahesh, Sheela Nampoothiri, Sindu T.K, and D. M. Vasudevan. Abstract No: 1.11: Enhanced 5HT2A Receptors in the Cerebral Cortex of Hypoxia Induced Neonatal Rats: Effect of Glucose and Oxygen Supplementation Anju T. R, Pretty Mary Abhraham and C. S. Paulose Abstract No: 1.12: Diagnosis of Organic Acidurias in Children - 2 Years Experience in a Tertiary Care Center M P Narayanan, Kannan Vaidyanathan and D M Vasudevan Abstract No: 1.13: Detection of Inborn Errors of Metabolism in Paediatric Age Group in a Tertiary Care Centre. Saritha KamathU, Licy C.D, Nalini Bhaskaranand, and Anjali Rao Abstract No: 1.14: Hyperprolinemia in 22q.11 Deletion Syndromes and its Association with Neurological Deficits Sweetha Mary Mammen, Dr. K.N. Subhakumari and M. V. Thampi Abstract No: 1.15: Hypoglycaemia Biswajit Saha Abstract No: 1.16: Dyslipidemias Prof. N. Haridas Abstract No: 1.17: Effect of Swimming on Selected Health Parameters of Beginners Ragini Srivastava, Royana Singh, Ratna Pandey and Amit Srivastava

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Abstract No: 1.18: Evidence-Based Guidelines for the use of Biochemical Markers of Bone Turnover in the selection and monitoring of Bisphosphonate Treatment in Osteoporosis: A Consensus Document of the Belgian Bone Club Dr.Maulik Nayak, Dr. Alok Parekh, Dr.Rita Shah and Dr. D.A. Trivedi Abstract No: 1.19: Requirement of Essential Fatty Acids: An Analytical Approach Dr. Sankar Roy Abstract No: 1.20: Clinical Approach to Integrated Basic Sciences Teaching Shruti Mohanty Abstract No: 1.21: A Study on Organic Aciduria: Detection and Quantification using HPLC Aswathi K.I, Narayanan MP and Kannan Vaidyanathan Abstract No: 1.22: ELISA based Evaluation of 17-á Hydroxy Progesterone as a Diagnostic Marker for Congenital Adrenal Hyperplasia and Assessment of Hyperhomocysteinemia in Coronary Artery Diseases and Cerebrovascular Accidents Jayakumar N., Dr. Sujaya P and Dr. Kannan Vaidyanathan Abstract No: 1.23: Standardisation of Chromatographic Techniques: HPLC & GC ­ A Diagnostic Tool for Inborn Errors of Metabolism Ms. Amalmol Peter and Dr. Kannan Vaidyanathan Abstract No: 1.24: Faecal Calprotectin in Inflammatory Bowel Diseases M.P. Narayanan, Kannan Vaidyanathan, O. Gajendra, S. Shine and V. Balakrishnan Abstract No: 1.25: A Study on Cytotoxicity and Inhibition of Cyclooxygenase-2 by Curcumin and Diallyl Sulfide in Head And Neck Cancer Cell Line Hariharasudan. R Abstract No: 1.26: A Study for Investigating the Association between Semen Biochemical Parameters and Routine Clinical Semen Parameters in Infertile Men Suresh S. and T. Vijayakumar Abstract No: 1.27: Isolation and Validation of the Biomarker MMP1 from Salivary Transcriptome for Early Detection of Oral Cancer Kannappan Alagappan, Amritha Suresh and M A Kuriakose Abstract No: 2.1: Serum Copper and Magnesium Level in Type ­ 2 Diabetes Mellitus B.K.Agrawal, Seema Tamrakar and V.K.Sharma

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Abstract No: 2.2: Biochemical Profile of Cord Blood in Normal Pregnancy and Pregnancy Induced Hypertension Miss D.Prathima , Dr. U.B.Vijaya lakshmi and Prof. Dr. K. Ambika Devi Abstract No.2.3: Relationship between Micronutrient Intake of Mothers and Metabolic Outcomes in Offspring (6 and 12 yrs): Pune Maternal Nutrition Study S.S.Naik, D.S.Bhat, C. Joglekar and C.S.Yajnik Abstract No: 2.4: Lack of Awareness Resulting in Lead Poisoning in Unorganized Sector of Battery Workers Dr. Vishal Babu G N, Mr. Raviraja A and Dr. T Venkatesh Abstract No: 2.5: Estimation of Zn, Mg in Several Protein Energy Malnutrition in Children Age Group 6 Month to 5 Year Farha Ali, B. K. Agrawal and Manjusa Goel Abstract No: 2.6: Zn and Fe Levels in Hypothyroid Subjects: A Hospital Based Case Control Study. Gaurav B. Shetty, Suchetha Kumari.N and Smitha.P Abstract No: 2.7: Extracellular Mg2+ Decreases Loading of the Ca2+-Sensitive Fluorescent Probe Chlortetracycline, into Calcium Stores in Lymphocytes and Neutrophils Nivedita L. Rao and Jose Jacob Abstract No: 2.8: Expression of Metallothionein (MT) in Colonic Precancerous and Cancerous Tissue in Animal Model Pamela Christudoss, and R. Selvakumar Abstract No: 2.9: A Study of Serum Ferritin Levels among Male Blood Donors Ramesh Kumar and Dr.Shanmugam Abstract No: 2.10: Study of Serum Magnesium Levels in Diabetic Nephropahty M. Ravi kanth, Dr. A.K. Bansal and Mrs. Lincy Jacob Abstract No: 2.11: Effect of Aluminum on Cardiac Rhythm of Male Albino Rats: An Electrocardiogram Study Sadhana Verma, Pradeep Kumar, M. Y. Khan, Sandeep Tripathi and A. A. Mahdi Abstract No: 2.12: Serum Calcium, Phosphorus and Alkaline Phosphatase Levels in Preterm Babies Sarika Singh Chauhan, B. K. Agrawal and Rashmi Dwivedi Abstract No: 2.13: Hypomagneseamia in Diabetes Mellitus Shymala Manga, Dr.B.Prabhakar Rao and Dr.Sunanda

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Abstract No: 2.14: Study of Biochemical Markers in Iron Deficiency Anemi

Smita B. Pawar

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Abstract No: 2.15: Estimation of Toxic Lead (Pb) Contamination of Tapioca (Manihot esculenta) in Kerala State Subin Joy, Shaikh Mohammed and V.V Pillay Abstract No: 2.16: The effect of Vitamin E Supplementation on Soluble P-Selectin Levels in Pre-Eclampsia Sujata Wangkheimayum, Sunil Kumar and Vanita Suri Abstract No: 2.17: 5 ­ Methyl Tetrahydrofolate in Dried Blood Spot as an Indicator of Folate Status in Humans Thippeswamy. T.G, Ravinder Punjal and Madhavan Nair.K Abstract No: 2.18: Age Related changes in Clcium, Phosphorous and Calcitonin in Healthy Women with special reference to Premenopausal and Post menopausal Women Miss V. Meenakshi, Dr.S.K. Sen and Dr. Roma Rattan Abstract No: 2.19: Effects of Resveratrol on Expression of Critical Bone Remodelling Factors in Surgical Menopause Model of Rat Alka Khera, Dr. Thungapathra M, Dr. Jaswinder Kalra and Dr. Sujata Ghosh Abstract No: 2.20: An Experimental Evaluation of Cytoprotective Effect of Amifostine on CHOP Chemotherapy in Albino Rats Ambili Remesh Abstract No.2.21: Biochemical Changes in Acute Organophosphorus Poisoning Indira A Hundekari, A.N.Suryakar, Nilima N. Dongre, J G Ambekar, B.B.Devaranavadagi and D.B.Rathi. Abstract No: 2.22: Lead Toxicity Among Automobile Workers Nilima N. Dongre, A.N.Suryakar, A.J.Patil,I.A.Hundekari, J.A.Patil,J.G.Ambekar B.B.Devaranavadgi, D.B.Rathi and Rama Jailkhani Abstract No: 2.23: Tacrolimus in Renal Transplant Recipients Dr.Pradeep Naik and Mallikarjuna .M Abstract No: 2.24: Biochemical Effects of Feeding Soft Drink and Ethanol Arun Raj, Praveen Kottath Veetil, Sheeba Varghese, J K Mukkadan and P K Joseph

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Abstract No: 2.25: Comparative Study for Slow Acetylator Status by Phenotype / Genotype Polymorphism of N-acetyltransferase-2 in Patients on Antitubercular Treatment S.V.Rana, Sanjeev K.Sharma, R. P. Ola, S.Arora, S.K.Sinha, P Pandhi and K.Singh Abstract No: 2.26: Correlation of Gall Stone Disease with IronDeficiency Anemia Prakash A; Lal A.K; Sachan P.K and Sahu S.K. Abstract No: 2.27: Trace Elements in Leprosy Dr V B Londhe, Sevarina Madhale, Dr Z G Badade, Dr P E Jagtap and Dr S K Ahaley Abstract No: 2.28: Oxidative Stress, Antioxidants and Trace Elements in Leprosy Dr V B Londhe, Sevarina Madhale, Dr Z G Badade, Dr P E Jagtap and Dr S K Ahaley Abstract No: 2.29: Variations in Serum Ferritin, Electrolytes and TIBC Levels in Paediatric Multitransfussed Thalassemic Patients Jaidev Singh, Dr. (Mrs) Meena Varma and Dr.Sangeeta Paneri Abstract No: 2.30: Study of Lead Content in Various Water Samples within Bangalore and A Survey Based on Effects of Lead Poisoning on People Preeti Mangala, Javeria Anjum and Arpi Dey Abstract No: 2.31: Evaluation of Lead Concentration in Milk and Milk Products Charanraj T.P, Anuj Ranjan, Alka Kumari, Arun Kumar, Kalpana Devi.N and Sushma Bharti Abstract No: 2.32: Effect of Lead on Employees Working in Paint Industry Hariprasad.B.N, Vinay.G, Vinay Kumar.C, Sathish Kumar.P..and..Prof.Sripathy.L Abstract no: 2.33: Vitamin D- Emerging Silent Lifestye Disease: A Pilot Study D. Radhika Chowdary and B. Naveen Kumar Abstract No: 3.1: Myeloperoxidase in Chronic Kidney Disease Usha Anand, Madhusudhana Rao.A, Vijaya D, Aruna V, Gayathri B and Anand CV Abstract No: 3.2: Serum Adenosine Deaminase Levels in Acute Kidney Injury Saleena Ummer V, Nalini K , Ravindra Prabhu and Licy CD

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Abstract No: 3.3: In vitro Studies on Anticalcifying Properties of achyranthes aspera Aggarwal A, Singla SK and Tandon C Abstract No: 3.4: Correlation of Serum Intact Parathormone Level with Biochemical Parameters in Various Stages of Chronic Renal Failure Pateints Bhimappa .N. M, Suma.M.N and H.S.Virupaksha Abstract No: 3.5: Comparison of Serum Creatinine, Urinary Creatinine and Urinary Beta 2 Microglobulin Levels as an Early Marker of Subtle Renal Dysfunction due to Aminoglycoside Therapy in Full Term Sick Neonates. Dr Bonny B Jasani, Dr Rohini Bhadre, Dr Varsha and D. Phadke Abstract No: 3.6: Observations on Microalbuminuria in Hypertensives with Alcholism and / or Smoking Dr.D.L.Lalitha, Dr.B.Sreehari babu, Mr.D.S.S.K.Raju and Dr.P.K.Behera Abstract No: 3.7: Serum Lipoprotein(a) and Lipid Profile in Chronic Kidney Disease Neela B M and Virupaksha H S Abstract No: 3.8: Evaluation of Urinary Protein Bound Sialic Acid and Protein Carbonyl content as the Prognostic Markers of Nephrotic Syndrome in Pediatric Age Group Niranjan G., Koner B.C and Bhat V. Abstract No: 3.9: Oxidative Stress and Transforming Growth Factor-Beta 1 (TGF-â1) in the Progression of Diabetic Nephropathy Kavitha Gandhi and Dr.Rita Mary Aruna Abstract No: 3.10: Hypothyroidism and Renal Impairment K.Shanthi Naidu, M.R.Chakravarthi, Bipin Kumar Sethi, Suryadeep Pratap, Syed Hyder Ali, BVLN Murthy and G.Surekha Abstract No: 3.11: Oxidative Stress and Antioxidant Therapy in Nephrotic Syndrome Jyoti Dwivedi and Dr.Purnima Dey Sarkar Abstract No: 3.12: Serum Cystatin C levels in Renal Transplant Recipients Krishnamurthy N , Usha Anand , Anand C V , Aruna V and Venu G Abstract No: 3.13: Serum Zinc and Serum Copper Level in Renal Diseases Payasvi Baweja, B.K. Agrawal and V.K Sharma

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Abstract No: 3.14: Renal Function Screening in Neonatal Asphyxia Prachi Paliwal, Dr.B.K.Agrawal, Dr.Rashmi Dwivedi and Dr.Paliwal Manoj Abstract No: 3.15: Exploring the Role of Lipids Isolated from Human Renal Stone Matrix in Urolithiasis Priyadarshini, Shrawan Kumar Singh and Chanderdeep Tandon Abstract No: 3.16: Prevalence and Characterization of Renal Tubular Acidosis in Patients with Myalgias, Osteoporosis and Mild Arthritis. Rachel Jacob Abstract No: 3.17: Renal Calculi Analysis K.Shanthi Naidu, Suryadeep Pratap, Zaheed Khan, Syed Hyder Ali, S.M.Ismail, and A.Phani Kumar, Abstract No: 3.18: Antioxidant Activity of Creatinine and Uric Acid and Related Efficacy of Antioxidant to Inhibit Glycation Rohit Suresh, Yogita S Phapale and Dr Z G Badade Abstract No: 3.19: Carbamylated Haemoglobin: A Diagnostic Marker for Chronic Renal Failure. S.Subramaniam, ShyamaSubramaniam, M.K. Mani, Partha Karmakar, K.Ezhil arasan, K.Dhananjayan and M.Gopinath Abstract No: 3.20: Evaluation of Low Dose Doxycycline on Renal Function in Diabetes with Nephropathy S. Seth, P. Talapatra, H.K. Aggarwal and M. Seth Abstract No: 3.21: The Effects of Kidney Function and Transplantation on Plasma Electrolytes, Albumin and Cardiovascular Risk Thompson, M.J.W. and Ray, U. Abstract No: 3.22: Tacrolimus in Renal Transplant Recipients Dr.Pradeep Naik and Mallikarjuna .M Abstract No: 3.23: Study of Microalbuminuria in Hypertension Sharma, Ashutosh. ; Lal, A.K. ; Mehrotra, V and Verma, S.K. Abstract No: 3.24: The Pattern of Nephrotic Syndrome in Malabar region, Kerala K Sreevidya and George Abraham Abstract No: 3.25: Adiponectin Levels in Chronic Kidney Disease and its Correlation with CRP, Cystatin-C and eGFR Manjunatha.G.J and Vijay S.Bhat Abstract No: 3.26: Effect of Smoking on Chronic Renal Failure in Primary Glomerular Nephropathies Prof.(Dr) Uday Kumar, Dr.A.Sharan and Dr Snigdha

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Abstract No: 3.27: The Effects of Kidney Function and Transplantation on Plasma Electrolytes, Albumin and Cardiovascular Risk Thompson, M.J.W. and Ray, U. Abstract No.3.28: Study of Urolithiasis in Gwalior Region A.K. Bhargava, Neelima Singh, Sanjeev Singh, Pradeep Sharma, Dharmveer Sharma, Sunil Agarwal and Sunita Sharma Abstract No: 4.1: Cardiovascular Risk Markers in Hypothyroidism Aparna R.Bitla, E. Jayanti, Alok Sachan, G. Shiva Krishna, M.M.Suchitra, P.V.L.N.Srinivasa Rao, and KVN Raju Abstract No: 4.2: Hyperhomocysteinemia, Circulating Folic acid and Cobalamine Levels in Coronary Syndrome Yadav AS, Bhagwat VR, Mane AY and Venugopal NT Abstract No: 4.3: Insulin Resistance and Lipid Profile in Pregnancy Induced Hypertension Asmathulla S, Rajagovindan.D and Jakanattane Abstract No: 4.4: Hypothyroidism Risk factor for Coronary Heart Disease B.K.Agrawal, Bhawna Bhimte, and V.K.Sharma Abstract No: 4.5: Diabetes Accelerates Age-Related Lipid Profile Disturbances Zafar M.E. Nehal M. Teeto S.A. and Prasad K.R. Abstract No: 4.6: Biochemical Study of Emerging Risk Markers in Coronary Heart Disease H.V. Singh, S.Bhandari, N.Singh, S.Singh and A.Raizada Abstract No: 4.7: Anti-oxidant Potential (AOP) of LDL in Post Menopausal Women Suffering from Type 2 Diabetes Mellitus. Nivedita Singh, Neelima Singh, Sanjeev Singh and Vishal Bhargava Abstract No: 4.8: Increased Levels of Inflammatory Markers, Sialic Acid and Cardiovascular Risk Factors in Type 2 Diabetes Mellitus. Dr. S.Rajesh, Dr. S.Sethupathy and Dr.S.M.Rajendran Abstract No: 4.9: Influence of Atherosclerosis and Exercise on Arterial Lipoprotein Lipase Activity in Pigs Shakti Aggarwal, Theodore W. Zderic, Perminder Gulani and Marc T. Hamilton, Abstract No: 4.10: Study of Apolipoprotein A-1, B and Lipid Profile in Premenopausal and Postmenopausal Women Dr. Swapnali and Dr. D.S. Jayaprakash Murthy.

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Abstract No: 4.11: Significance of Plasma Total Homocysteine Levels in Type II Diabetes Mellitus Ppatients Anbazhagan M and Ben S. Ashok Abstract No: 4.12: Apolipoprotein A1 Gene Polymorphism (G75A and C+83T) in Patients of Myocardial Infarction- a Pilot Study in North Indian Population. Rajni Dawar, Anil Gurtoo and Ritu Singh Abstract No: 4.13: Role of Pro-inflammatory Cytokines, ApoB and Small Dense LDL in Women with Coronary Artery Disease Sunita, S.B Sharma, D.Puri, R.L.Tripathi and S.Dwivedi Abstract No: 4.14: Dyslipidemia and Anti-oxidant Nutrient in Preeclampsia Dr.M.G.Dhabe, Dr. (Mrs.)S.C.Choudhary and Dr.M.R.Mogarekar Abstract No: 4.15: Post Coronary Stenting Decrease in Atherogenic Risk Factors Kavitha.S, Sridhar M.G, Satheesh.S and Balachander J Abstract No: 4.16: Serum Uric Acid in Acute Myocardial Infarction Gargi Sen and Virupaksha H. S. Abstract No: 4.17: Apolipoprotein B, Total Cholesterol, HDL-C, Non HDL-C, LDL-C and Triglycerides as Risk Factors of Atherosclerotic Vascular Disease Jitty George, Dr.Andrews M.A, Dr.Cibu Mathew, Dr.Geetha Damodaran K, and Dr.Gilsa E Abstract No: 4.18: Accuracy of Retinal Changes in Predicting Microalbuminuria in Obese and Non-obese Hypertensive Patients. Arul Senghor, Ebenezer William and Jothimalar Abstract No: 4.19: Circulating Levels of Cell Adhesion Molecules in Hypertension Kavita K. Shalia, Manoj R. Mashru, Jagdish B. Vasvani, Rajashree A. Mokal. Shilpa M. Mithbawkar and Priti K. Thakur Abstract No.4.20: Serum Paraoxanase Activity and its relationship with Atherogenic Index of Plasma (AIP) in Hyperlipidemic Patients Kavya R, Phani M.N, Supriya M, Jeevan K.Shetty and Mungli Prakash Abstract No: 4.21: Trends in Blood Pressure with Increasing Serum Homocysteine Levels Kumar. Animesh, Gupta P.R and Ali.Z Abstract No: 4.22: Coronary Heart Diseases and Serum Triglycerides K Ram achandran, Divya K.V., Ajeena and Sheela

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Abstract No: 5.1: Investigation Profile Correlated with Adverse Maternal and Perinatal Outcome in Pregnancy Induced Hypertension (PIH) Ms.Liggy Andrews, Dr.N.Haridas and Dr. Smruti Vaishnav Abstract No: 5.2: Status of Lipid Profile in Patients of Acute Myocardial Infarction K.Suchitra, Bari Siddiqui M. A and U.Satyanarayana Abstract No: 5.3: A Study of Age Related Changes in the Serum Lipid Levels in Healthy Adults of Mangalore City Dr.Malathi.M and Ms.Sangita Kamath Abstract No: 5.4: Systemic Failure of Interferon-á synthesis, a Potent Inhibitor of Platelet Aggregation and a Thrombolytic Agent, and its Role in the Development of Acute Myocardial Infarction. Mau Bhattacharyya and Asru K. Sinha Abstract No: 5.5: Atherogenic Index in Patients with Established Myocardial Infarction. Abdul Lateef , Afshan Rafi and Pragna Rao Abstract No: 5.6: Increased Cardio Metabolic Risk Factors seen in School Cildren Sumathi, Meera, Vinodhini and Ebenezer William Abstract No: 5.7: Assessing Risk Factors in Acute Coronary Syndrome Moushumi Lodh, Ashok Parida and Alpana Saxena Abstract No: 5.8: Atherogenic Index of Plasma in Ischaemic Heart Disease Patients. Phani M.N, Supriya.M, Kavya R, Jeevan K.Shetty and Mungli Prakash Abstract No: 5.9: Current Biomarkers for Myocardial Infarction Rathish.R and Sumathi.P Abstract No: 5.10: Appearance of Dermcidin, a Heat Shock Protein, as a Risk Factor for Acute Ischemic Heart Disease through the Inhibition of Pancreatic Insulin Synthesis Rajeshwary Ghosh and Asru K.Sinha Abstract No: 5.11: Uric Acid Levels in Cardiovascular Disease Mrs. Ranjita V.Gaur and Dr.(Mrs.)Rita M.Shah Abstract No: 5.12: Effect of a Single Dialysis Session on Plasma Lp(a) Levels in Patients on Maintenance Haemodialysis Seshadri Reddy. V, M.M. Suchitra, E. Prabhakar Reddy, Aparna R.Bitla, V. Sivakumar and P.V.L.N. Srinivasa Rao

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Abstract No: 5.13: Association of LDL Particle Size with other Cardiovascular Risk Markers in an Industrial Population. Mohammad Tarik, Lakshmy Ramakrishnan, Ruby Gupta, Dorairaj Prabhakaran and Kolli Srinath Reddy Abstract No: 5.14: A Novel Antihypertensive Protein Hormone from the Kidney Cortex, and its use for the Diagnosis of Essential Hypertension in Men. Somashree Chakraborty, Gausal A. Khan and Sinha A.K Abstract No: 5.15: Ischemia Modified Albumin ­ an Evolving Diagnostic Marker of Myocardial Ischemia Dr Sudeshna Behera and Prof. P.C.Mohapatra Abstract No: 5.16: Study of Serum Magnesium, HbA1c , Lipid Profile and Microalbuminuria in Diabetic Retinopathy Swetha N K and Virupaksha H S. Abstract No: 5.17: The Effects of Kidney Function and Transplantation on Biochemical Correlates of Cardiovascular Health Thompson. M.J.W and Ray.U. Abstract No: 5.18: Utility of NT proBNP in a Wider Range K.Shanthi Naidu, Suryadeep Pratap, Syed Hyder Ali, BVLN Murthy, B.Goud, B.Raghavendra and J.Visalasree Abstract No: 5.19: Normalization by Aspirin of Hypertension for the Reduction of Risk of Coronary Artery Disease in Men Uttam K. Maji and Asru K. Sinha Abstract No: 5.20: A Study of Insulin Levels in Pregnancy Induced Hypertension Vinodhini, Meera Shivshekar and Ebenezer William Abstract No: 5.21: Levels of GHBA1C in Non- diabetic Patients with Cardiovascular Disease Tarun Kumar Sharma, H. Parchwani, G.G. Kaushik and Vijay Shankar Abstract No: 5.22:Biochemical Markers and Coronary Occlusion in Patients Undergone Coronary Arteriography Bhagwat V.R, Venugopal N.T, Yadav A.S and Mane A.Y Abstract No: 5.23: Cytomegalovirus and Coronary Heart Disease Asif.A. Abstract No: 5.24: Role of "Ischemia Modified Albumin" in the Early Diagnosis of Acute Coronary Syndromes B.Gayathri, D.Vijaya, B.Bini, Usha Anand, V. Aruna and C.V Anand

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Abstract No: 5.25: Inflammatory Markers in Arterial Prehypertension and Hypertension Manjrekar AP, D'Souza V, Hegde A, Adhikari P and CV Raghuveer Abstract No: 6.1: A Spectrum of Paraproteinemias K.Shanthi Naidu, P.Srikanth, Shailesh.S, M.R.Chakravarthy, Syed Omar, Kishore, A.Suhasini and Visalasree Abstract No: 6.2: Effective POCT : the Need for a Broader Perspective A.K.Srivastava Abstract No: 6.3: Biochemical Markers in Osteoporotic Hip Fractures Dr.Meera.S , Dr.M.Sukharaja , Dr.B.G.Sagar and N.Kantharaj Abstract No: 6.4: Metabolic Dearrengements in Births Asphyxia Occurring as a Result of Cellular Injury. Amrita Vamne, B. K. Agrawal and Jyotsana Shrivastava Abstract No: 6.5: Evaluation of Salivary Cholinesterase and Ceruloplasmin in Children with Attention Deficit Hyperactivity Disorder Bhavya Prabhu.K, Archana E, Priya Pai, Dr. Krishnananda Prabhu and Dr. Anjali Rao Abstract No: 6.6: Cheek Cell Epithelium: a Non Invasive Specimen for Measurement of Fatty Acid Intake Ransi Ann Abraham, Lakshmy Ramakrishnan ,Ruby Gupta, Smita Golandanz, Rajinder Parshad, V K Bahl, Ambuj Roy, R M Pandey, Dorairaj Prabhakaran and Kolli Srinath Reddy Abstract No: 6.7: Coagulation Deficiencies in a Referral Laboratory K.Shanthi Naidu, P.Srikanth, Shailesh.S, D.Ramesh, D.Ramana, B.Raghavendra, B.Goud and B.Siva Mallika Abstract No: 6.8: Coagulation Studies in Neonatal Critical Care K.Shanthi Naidu, P.Srikanth, Shailesh.S, D.Ramesh, D.Ramana, B.Raghavendra, B.Goud, B.Siva Mallika and S.M.Ismail Abstract No: 6.9: A Case of Xanthochromia Ashish Sharma, N. Haridas, S. Ghosh, Hitesh Shah ,Kiran and Uday Abstract No: 6.10

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Abstract No: 6.11: The Potential Significance of Procalcitonin and Interleukin-6 in Prediction of Survival in Patients with Sepsis S. Das, S. Bhargava, A. Manocha, M. Kankra and L.M.Srivastava Abstract No: 6.12: Immunological Studies on Glycated and Glycoxidated LDL: Implications in the Pathogenesis of Various Diseases Salman K A, Alam.R, Arif.N and Khan.S Abstract No: 6.13: Critical Call Back ­ An Experience in a Tertiary Care Hospital Dr. Mohammad Ibrahim, Maj Dr. Ikramul Islam (Retd), Prof (Dr.) Tareak-Al-Nasir and Lt.Col Dr. Debashish Saha Abstract No: 6.14: Rare cases of Pagets Disease of the Bone in India: Biochemical and Clinical Evaluation and Follow-up in 2 Extreme Presentations Dr CS Muralidhara Krishna ,Dr H L Vishwanath ,Dr S Sanjay, Dr Tejeswini Deepak, Dr.A Sharda and Dr.S.S Srikanta Abstract No: 6.15: Role of Acute Phase Proteins in the Prognosis of Head Injury P Sumathi and M K Balasubramanian Abstract No: 6.16: Biochemical Profile of Cord Blood in Normal Pregnancy and Pregnancy Induced Hypertension. Miss D.Prathima , Dr. U.B.Vijaya lakshmi and Prof. Dr. K. Ambika Devi Abstract No: 6.17: Comparision of Plasma Phosphdiesterase Activity in Premenstrual and Menstrual Period. Rajyalakshmi M, Indu R Warrier, Dr. Krishnananda Prabhu and Dr. Anjali Rao Abstract No: 6.18: Serum Calcium, Phosphorus and Alkaline Phosphatase Levels in Preterm Babies. Sarika Singh Chauhan, B. K. Agrawal and Rashmi Dwivedi Abstract No: 6.19: Study of Effect of Phototherapy on Neonatal Hyperbilirubinemia M.Vijay and M.Malathi. Abstract No: 6.20: SMS Tube : A Novel Way of Giving Specimen Collectors 24/7 Mobile Access to Our Specimen Requirements Database T F Hartley, W. Bowling, Sharon Fong, A, Gibb, K Hen, B Hitchens, Renee Twynam-Perkins, K Wilson, J Zhang and Nicole Herbert

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Abstract No: 6.21: Storage Stability of Glycosylated Hemoglobin in Red Blood Cells and Dried Blood Spots Ruby Gupta, Ramakrishnan Lakshmy, Dorairaj Prabhakaran and Kolli Srinath Reddy Abstract No: 6.22: Data Analysis of Biochemical Values in a Polyclinic and Diagnostic Centre in Dumdum, Kolkata Dr.Aditi Sen (Sarkar), Sanchari Halder, Anirban Ganguly, Mononjay Banerjee, Ramkrishna Bhattacherjee and Dr. Papia Mukherjee Abstract No: 6.23: Evaluation of Salivary Magnesium and Protein Thiols in Children with ADHD Archana E, Bhavya Prabhu K, Priya Pai, Dr. Krishnananda Prabhu MD, Dr. Anjali Rao and Dr. Revathi P. Shenoy Abstract No: 6.24: Estimation of Creatinine in Sweat Shivaraj Gowda, Prakash Desai, Avinash A K Math, Sonal Vernekar, Shruthi Kulkarni, Vinayak Hull and Mahantesh Ilkal Abstract No: 6.25: Reference Range for Serum Total Alkaline Phosphatase in Healthy Individuals Kiran Kumar B , Nalini Ganesan and Anbazhagan M Abstract No: 6.26: Quality Control Evaluation using Quality Indicators - Experience of a Clinical Laboratory in Delhi A Manocha, S Bhargava, M Kankra, S Das and LM Srivastava Abstract No: 6.27: Quality Indicators for Measuring the Process Performance of a Clinical Laboratory Dr.R.Balambal Abstract No: 6.28: Serum Protein Electrophoretic Pattern of Chronically ill Patients in a Tertiary Care Hospital Vijayasree.N Abstract No: 6.29: Immunocharacterisation of Polyclonal and Monoclonal Antibodies Raised against Cobra Venom Toxin Lakshmi.A , Dr. K.S.Vasudevan, Mr.A.Mathialagan and Dr.S.Subramanian Abstract No: 6.30: Development of a Novel Method to Eliminate the Negative Interference of Bilirubin on Creatinine Assay by Jaffe's Kinetic Method in Icteric Sera. M.M.Goyal and A.Basak Abstract No: 6.31: Clinical Biochemistry : The Goldmine of Challenges and How do we face it K. Sorte, K. Palandurkar, M.M. Goyal and A.Basak

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Abstract No: 6.32: Hyperhomocysteinemia and Folic Acid, Vitamin B12 Levels and their Correlation in Eclampsia Shahid A. Mujawar, Vinayak W.Patil, Rekha G. Daver and Narayan A. Madrasi Abstract No: 7.1: The Association between Serum Alkaline Phosphatase and ABO Blood Group E.Sruti, Nalini Ganesan and Anbazhagan.M Abstract No: 7.2: Guidelines and Recommendation for Laboratory Analysis in the Diagnosis Dr. Alok Parekh, Dr.Maulik Nayak, Dr.Rita Shah and Dr. D.A. Trivedi Abstract No: 7.3: Establishing a Reference Range for Thiopurine Methyltransferase (TPMT) Activity in Indian Populations Jayashree.T, Sadik A. Mahammad, Ramchandra Rao.O, Rupa Banerjee and Nageshwar Reddy.D Abstract No: 7.4: Biochemical Impact of Lumirubin in Neonatal Hyperbilirubinemia P.S Kamble, R.K Padalkar, P.D Zende and V.R Pandhare Abstract No: 7.5: Comparison of Cord Blood Atherogenic Index in Males and Females Kharb.S and Smiti Nanda Abstract No: 7.6: Quality Control in Clinical Chemistry: Minimal Requirements for an Acceptable Program Dr. Manju Bala Pahwa and Dr. Veena Singh Abstract No: 7.7: Quality Indicators: A Tool for Continuous Quality Improvement in Clinical Laboratories S.Subramaniam and Shyama Subramaniam Abstract No: 7.8: Serum Sodium and Potassium Level in Emesis Gravidarum Sana Hasan, Dr. B.K.Agarwal and Dr. Neelkamal Kapoor Abstract No: 7.9: Study of Serum Fluoride and Sialic Acid Levels in Osteosarcoma Sandhu.R, Lal.H, Kundu Z.S and Sandeep, Kharb.S Abstract No: 7.10: Prevalence of Diabetes Mellitus in Urban Region of Chennai City- a part of Purse- HIS study Satyajeet Giri, S.Thanikachalam and K.Sowmya Abstract No: 7.11: A Study of Serum Ferritin Levels in Type -2 Diabetic Patients Shiv Ram Krishn and Ekta Agarwal

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Abstract No: 7.12: Comparison of Serum and Plasma Uric Acid Dr. Shivraj Gowda, Dr. P.B. Desai, Shruti Kulkarni, Sonal Vernekar, Avinash A.K.Math, Vinayak Hull and Mahantesh Ilkal Abstract No: 7.13: Tourniquet: Lacunae of Preanalytics Smita S.Sonoli, and Anuradha B.Patil Abstract No: 7.14: Simplified Procedure for Cholesterol Determination Dr. Shivraj Gowda, Dr. P.B. Desai, Sonal N. Vernekar, Vinayak Hull, Avinash A.K.Math, Shruti Kulkarni and Mahantesh Ilkal Abstract No: 7.15: Study of Lipid Profile and Glycosylated Hemoglobin (HbA1 C) in Patients of Diabetes Mellitus Tripti Saxena, B.K. Agrawal, and V.K. Sharma, Abstract No: 7.16: Use of Tris-Borate Buffer in Agarose Gel Electrophoresis of Serum Proteins Mahesh S, Naureen Anwar, Yeshoda K, Saifa Yasmin and Dr. Nalini K Abstract No: 7.17: Use of the Laboratory in Prediction of Outcome in the High-Risk Newborn Dr. Alok Parekh, Dr.Maulik Nayak, Dr.Rita Shah and Dr. D.A. Trivedi Abstract No: 7.18: Standardization of DAM Method by Single Step Dr. Shivraj Gowda, Dr. P.B. Desai, Vinayak Hull, Avinash A.K.Math, Shruti Kulkarni, Sonal Vernekar and Mahantesh Ilkal. Abstract No: 7.19: Effect of Feeding Practices in Neonatal Hyperbilirubinemia Subjects P.D Zende, R.K Padalkar, P.S Kamble and V.R Pandhare Abstract No: 7.20: Novel Therapeutic Properties of Nano Silver* Siddhartha Shrivastava, Sunil Singh and Debabrata Dash Abstract No: 7.21: Evaluation of Procalcitonin as a Diagnostic Marker of Sepsis Ms. Nandini C and Dr.N.Leela Menon Abstract No: 7.22: Comparison of Various Parameters with Serum Free Light Chain Assay in the Diagnosis of Multiple Myeloma Mrudula E.V and Dr.Sajitha Krishnan Abstract No: 7.23: Importance of Tacrolimus Measurement and Assessment of its Toxicity in Monitoring Post Liver Transplant Patients Anuja.P.S, Dr. K.N. Subhakumari and Dr. Jagathlal.P.C Abstract No: 7.24: Six Sigma Analysis of Performance of Routine Chemistries at a Hospital lab Rashmi and Dr. Narayani

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Abstract No: 8.1: Hepatitis B Virus Sequence Database for Indian Isolates and Mutation Profiling Tool Amit Tuteja, Kaushal Madan, Subraya K. Acharya, Dr. K.Narayanasamy and Dr. Swati Subodh Abstract No: 8.2: Association of Antioxidant Enzyme Gene Polymorphism in ATT Induced Hepatotoxicity in North Indians S.V. Rana, Sanjeev K. Sharma, R.P. Ola, S.K. Sinha, J. Kaur, and K. Singh. Abstract No: 8.3: Circulating Plasma DNA as a Diagnostic Tool ­ A Review K.Ramadevi Abstract No: 8.4: Polymorphism of p53 Gene Codon 72 in Indian Hyperlipedimic Patients with Cardiovascular Disease K Ezhilarasan, K Dhananjayan, K.Anbarasu, A Sampath kumar, A K.Munirajan, Shyma Subramaniam and S.Subramaniam Abstract No: 8.5: Silver Nanoparticles - Potential Antiplatelet / Antithrombotic Agents Debapriya Bandyopadhyay and B.K Gupta Abstract No: 8.6: Rapid Detection of Mutation in RRDR of rpo B Gene for Rifampicin Resistance in MDR-Pulmonary Tuberculosis by DNA Sequencing Dr Surajeet Kumar Patra, Dr Anju Jain, Dr B L Sherwal and Dr Ashwani Khanna Abstract No: 8.7: Apo E Genotyping from Blood Stored on Filter Paper Rizwana Quraishi, Ramakrishnan Lakshmy, Ashok K. Mukhopadhyay and Bansi L Jailkhani Abstract No: 8.8: Angiotensin Converting Enzyme Gene Insertion Deletion (I/D) Polymorphism and its Association with Microalbuminuria in Essential Hypertension K.Ramalingam , K.Santha , S.Sethupathy and R.Vinoth Kumar Abstract No: 8.9: Association of Antioxidant Enzyme Gene Polymorphism in ATT Induced Hepatotoxicity in North Indians S.V. Rana, Sanjeev K. Sharma, R.P. Ola, S.K. Sinha, J. Kaur and K. Singh. Abstract No: 8.10: Expression of Myogenin and CD105 in Autologous Bone Marrow Derived Mesenchymal Stromal Cells of Duchenne Muscular Dystrophy Patients. Dr. Sumit Jhajharia, Dr Prakash C.Mohapatra, Debashish Das and Sujay Singh

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Abstract No: 8.11: Somatic DNA Damages in Cardiovascular Autonomic Neuropathy Supriya Simon A, Jayapal V and Dinesh Roy D Abstract No: 8.12: Interaction of Iron Deficiency Anaemia and Alpha Thalassaemia in the Population Tapash Rudra, Sila Chakrabarti and Bani Sengupta Abstract No: 8.13: Potential of Mycobacterial Excretory Secretory Protein Antigens (SEVA TB ES-31, ES-43, EST-6 and ES-20) as Biomarkers to detect Mycobacterium tuberculosis Bacilli Anindita M, Thamke.D, Mendiratta D.K and Harinath B.C Abstract No: 8.14: Reliability of Pleural Fluid ADA as a Marker for Exudates Mr.D.S.S.K.Raju, Dr.D.L.Lalitha, Dr.B.Sreehari Babu and Mr.M.Anil Kumar Abstract No: 8.15: Computational Studies of the Interactions of Some Inhibitors with HIV-1 Protease Rajkumari Rathore, Savita Rathore, Neha Sharma and Anil Bidwai Abstract No: 8.16: Auto Antibody in The Cases Of Neurological Disorder Dr.Anand Saran,Mr.S.S.Haque, Dr.Ashok Kumar and Dr.Uday Kumar Abstract No: 8.17 Evaluation of Serum Adenosine Deaminase Activity and Lipid Profile in Type 2-Diabetes Mellitus M.M.Suchitra, S.Aruna, K.V.N.Raju, K.Deepthi, V.Suresh, Aparna R.Bitla and P.V.L.N.Srinivasa Rao Abstract No: 8.18: Immunodiagnostic Evaluation of Recombinant Filarial Antigen rWbL2. Mahendra B.Gandhe, B.V.Sivaprasad.,Kiran Pote and M.V.R.Reddy Abstract No: 8.19: Immunomodulatory and Respiratory Burst Killing Activity of Pyrazine-2-Carbohydrazide Derivatives P.B.Miniyar and S. J. Makhija Abstract No: 8.20: Serum Lactate ­ Not a Good Marker of Disease Severity in Adults With Plasmodium Falciparum Infection Dr.Rina Triparthy and Dr.Bidyut Kumar Das Abstract No: 8.21: Altered Renal Profile and Liver Function Test in Case of Complicated Plasmodium Falciparum Malaria Shivaraj Gowda, Prakash B Desai, Sindhu J Shetty, Vijayetha S Kagwad and Mahantesh B. Ilakal

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Abstract No: 8.22: A Study on Acute Phase Proteins and its Relationship with Hemoglobin in Anemic Pulmonary Tuberculosis Patients Subodh Kumar, Kiran Saxena, Rashmi A Kulkarni, Avanish Mishra, Ramesh Pradhan, Rosy L.Pradhan and A.R.S Sinha Abstract No: 8.23: Plasma levels of Interleukin-6 and Interleukin10 in Rheumatoid Arthritis Vasanthi Pallinti, Nalini Ganesan and Rajasekhar.G Abstract No: 8.24: Anti - CCP Auto Antibodies in Rheumatoid Arthritis Vinod AN, Nalini Ganesan and Rajasekar Abstract No: 8.25: Molecular Mechanisms Underlying the Effect of Papaya Seed Products in Regulating Male Fertility N. Pathak, R. M. Samartha, D. Jain, K. K. Maudar, S. Tiwari, S. Gupta and P. K. Mishra Abstract No: 8.26: Prevalence of HCV Genotypes and Significance of TH1/TH2 Cytokines in Response to Combination Therapy: A Pilot Study from Central India P. K. Mishra, A. Bhargava, R. P. Punde, S. Tiwari, P. Vashistha, S. Varshney and K. K. Maudar Abstract No: 8.27: Diagnosis of Mycobacterial Tuberculosis Complex (MTBC) in Endoscopic Biopsy Samples and ParaffinEmbedded Tissue Specimens Using Real Time PCR: Implications in Patient Care G.V. Raghuram, A. Bhargava, R.P. Punde, S.Varshney, K.K. Maudar and P. K. Mishra Abstract No: 8.28: Mediators of Immune System and Their Possible Role in Pathogenesis of Chronic Hepatitis B and C Viral Infections R. P. Punde, A. Bhargava, S. Gupta, S. Varshney, P. Vashistha, K. K. Maudar and P. K. Mishra Abstract No: 8.29: Electrophoresis and Immunochemical Quantitation of Serum IgG, IgA, and IgM in Multiple Myeloma and Study of Complications in Multiple Myeloma Dr. Sajitha Krishnan and Toni Thomas Abstract No: 8.30: Comparative Evaluation of Recombinant WbSXP-1 Antigen and B.Malayi Microfilarial Antigens in the Detection and Management of Bancroftian Filariasis Partha Sarathi Pal, Gajalakshmi Dakshinamoorthy, Sivaprasad BV and M.V.R Reddy

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Abstract No: 8.31: Oxidalitive Stress and Immunity Status in Leprosy V.B.Londhe, Sevesina P. Madhale, Z.G..Badade, P.E Jagtap, S.K Ahaley and M.M Chavan Abstract No: 8.32: Real Time PCR Quantitation of Minimal Residual Disease (MRD) in Acute Lymphoblastic Leukemia (ALL). K.J Suneetha, K.Nirmala Nancy, K.R Rajalekshmy, T.G Sagar and T.Rajkumar Abstract No.8.33: Role of GST Polymorphisms in the Susceptibility and Outcome in Acute Lymphoblastic Leukemia K.J Suneetha, K.Nirmala Nancy, K.R Rajalekshmy, T.G Sagar and T.Rajkumar Abstract No: 9.1: CSF Calcium and Magnesium (Ionic and Total) Levels: a Preliminary Study Asharani.N, Sindhu, Samson Sujit Kumar.G, Ramakrishna and Ramalingam.K Abstract No: 9.2: Serotonin, Dopamine and Norepinephrine Functional Regulation of DA D1 Receptors in Unilateral Rotenone lesioned Parkinson's Rat Model. Jes Paul, Nandhu M.S, Korah P. Kuruvilla and C.S.Paulose Abstract No: 9.3: Dopamine D1 Receptor Up Regulation In Cerebellum and Brain Stem In Unilateral 6-Hydroxy Dopamine Rat Model: Antagonism By Serotonin And Gamma Amino Butyric Acid Nandhu. M. S, Jes Paul, Korah P Kuruvilla and C S Paulose* Abstract No: 9.4: Status of Enzymatic and Non-Enzymatic AntiOxidant Systems in Patients with Acute Ischemic Stroke J. Sudha Rani Abstract No: 9.5: Nitric Oxide and Lipid Profile in Type II Diabetes Mellitus-- A Case Control Study in East Sikkim Dr. A. Ghosh, Ms. Y. Bhutia, Dr. T .A. Singh and Dr. M.L Sherpa Abstract No: 9.6: Blood Glucose in Small for Gestational Age Infants Arvindkumar Chowdhary, R Nisarga, C.S Muralidhara Krishna and K.L Mahadevappa Abstract No: 9.7: Insulin Resistance and Lipid Profile in Pregnancy Induced Hypertension Asmathulla.S and Rajagovindan D. Jakanattane

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Abstract No: 9.8: Thyroid Satus in Hspital Bsed Cses from Rajendra Institute of Medical Sciences, Ranchi Dr K. K. Sinha and Dr. B. R. Ekka Abstract No: 9.9: Effect of Iron Deficiency Anemia on Glycosylated Hemoglobin Levels in Type 2 Diabetics. Dr.B.Shanthi M.D Abstract No: 9.10: Minerals Status in Thyroid Disorders (Hypo & Hyper thyroidism) B.Suneel, D.R.Nagendra, K.Ramalingam and J.N.Naidu Abstract No: 9.11: The Role of Salacia Oblonga in Maintaining Glucose and Lipid Homeostasis in Diabetic Rats Bhagyajyothi.D , Dr.Vivian D'Souza , Dr.Poornima Manjrekar and Dr. C.V. Raghuveer. Abstract No: 9.12: Glycemic Control and Complications of Diabetes Mellitus Adiga Usha and Adiga M.N.S Abstract No: 9.13: Relationship of Staple Diet and Abnormal Thyroid Function Observed in Outpatients at Ujjain (MP) Mane A. Y., Bhagwat V. R., Potey G.G and Gaikwad S.B Abstract No: 9.14 Evaluation of Cardiovascular Risk Markers in First Degree Relatives of Type 2 Diabetes Mellitus H. Nandeesha, V. Sathiyapriya, Zachariah Bobby, Aparna Agrawal and N. Selvaraj Abstract No: 9.15: Lipid Profile and Glycemic Status in Nonsmoking Type 2 Diabetes Mellitus on Treatment with Oral Hypoglycemic Drugs J. Sandhya, B.R.Shyam Prasad, and U.Satyanarayana Abstract No: 9.16: Thyroid, Lipid and Glycemic Profile in Dyslipidemia and Altered Thyroid Status Dr.Sridevi , Dr.Vivian D'souza, Dr.Poornima Manjrekar and Dr.Vinit Anand Abstract No: 9.17: Incontrovertible Behavior of Oxidative Stress in Etiopathogenesis of Diabetes Mellitus or its Complications is Disputable P.P. Singh, Anu Chandra, Chetna Vashishtha, Farzana Mahdi and Abbas Ali Mahdi1 Abstract No: 9.18: A Comparative Analysis of Anthropometric Measurements, Lipid Profile, Fasting Insulin and HOMA2-IR to Identify Insulin Resistance in Overweight Individuals Dr.M.D.Sreenivasa Murthy, Dr. Manjunath M.Tembad and Dr. Jayaprakash Murthy D.S

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Abstract No: 9.19: Thyroid Hormone Levels in New Borns of Mothers with Gestational Diabetes Dr.Geetha A, Dr. Joseph P. Thyparambil, Dr. Sreekumari.S and Dr.Sulekha.B Abstract No: 9.20: Lp(a) in Hypothyroidism F.S.Geethanjali, M.S. Seashadri and A.S. Kanagasabapathy Abstract No: 9.21: Role of Myelin Oligodendrocyte Glycoprotein and Galactocerebroside Antibodies on Phosphorylation and Degradation of Myelin Basic Protein In Isolated Myelin: Implications in Multiple Sclerosis. Krishnakumar Menon, Sidhy Viha C.V, Krishna Prasad G.N. and Claude C.A. Bernard Abstract No: 10.1: Glycosylated Hemoglobin Levels in Relation to Antioxidant Status and Lipid Profile in Type 2 Diabetes Mellitus Patients Prasiddha Tilak, Mahesh S, Naureen Anwar and Dr Mungli Prakash Abstract No: 10.2: Lipid Indices in Type 2 Diabetes Mellitus and Their Association with Macro and Micro-vascular Complications Dr. Imran Ahmed Siddiqui , Dr. J Rama Rao and Dr. Prabhavati Modi Abstract No: 10.3: Evaluation of Prothrombin Time and Activated Partial Thromboplastin Time in Type-2 Diabetes. Dr. Itishri Jena and Prof. Prakash Ch. Mohapatra Abstract No: 10.4: Effect of Trigonella Foenum Graecum and Insulin on Altered Membrane Functions in Alloxan Diabetic Rat Brains Pardeep Kumar, Asia Taha, R.K. Kale and Najma Zaheer Baquer Abstract No: 10.5: Metabolic Syndrome in Type 2 Diabetes Mellitus Priya.K.Dhas, Janet, Shantharam and Dr. Rita Mary Aruna Abstract No: 10.6: Study of the Effects of HCN Exposure in Cassava Workers Priya.K.Dhas, P.Chitra and Dr. Rita Mary Aruna Abstract No: 10.7: Prevalence of Obesity in Female Students Residing in Hostels in the University Campus of Manipal Rosemol Jacob M, Yeshoda K, Elsa Maria, Uma Pallavi, Asha Kamath, Dr. Revathy P Shenoy and Dr. Anjali Rao Abstract No: 10.8: In vitro Effect of Visfatin on Glucose Utilization by Visceral Adipose Tissue Radhika G, Magnus J, Sasikala M, Venkat Rao G, Pradeep Rabella, and Nageshwar Reddy.D

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Abstract No: 10.9: Salivary Cortisol Level in Depression Patients Sukanya Shetty, Suhas Pattar, Satheesh Rao and Srinivas Bhat Abstract No: 10.10: Atherogenic Index of Plasma and its Relationship with Fasting Blood Glucose in Diabetes Mellitus Patients. Supriya M, Kavya R, Phani M.N, .Jeevan K Shetty and Mungli Prakash Abstract No: 10.11: Lipoprotein (a), Fibrinogen, C ­ Reactive Protein, Uric Acid and Microalbuminuria in Coronary Artery Disease and Diabetes Mellitus" Suvarna.T. Jadhav, Dr. A.V. Sontakke, Dr. B.M. Tiwale and Dr. Mrs. C.C.Khanwelkar Abstract No: 10.12: Is hsCRP an Independent Risk Factor in Type 2 Diabetes? Dr.V.S. Kalaiselvi Abstract No: 10.13: Oxidative Stress and Proteolytic Activity in Erythrocytes of Diabetic Patients Dr. Varashree B S and Dr.P.Gopalakrishna Bhat Abstract No: 10.14: Leptin Levels in Early Post Transplant Period: Can it be Used to Assess Changes in Body Composition ? Victoria Job, Venkataramana Raju and George T. John Abstract No: 10.15: Inhibition of Type I 5á-reductase Vijaya Patil, Debjani Dasgupta,, Grace Samual, Shubhangi Mirapurkar and Krishnamohan Abstract No: 10.16: Thyroid Function in Diabetic Subjects Ranjani.P and Vijaya Srinivasan Abstract No: 10.17: Atypical Findings of Oral Glucose Tolerance Test (OGTT) Curve in Gestational Diabetes Mellitus (GDM) Shivaraj Gowda, P B Desai, Vijayetha S Kagwad, Sindhu J Shetty and M B Ilakal. Abstract No: 10.18: Insulin Level in Obese Patients with Obstructive Sleep Apnea Syndrome. Yogendra Singh, Surya Prakash Bhatt, Randeep Guleria and Manjit Singh Dhillo Abstract No: 10.19: Study of Lipid Profile, Oxidative Stress and Enzymatic Antioxidant in Type-2 Diabetes Mellitus Youb Raj Neupane, Dr Sandeep Roy and Dr Z G Badade Abstract No: 10.20: Assay of CRP and HbA1C in Smokers and Nonsmokers with Type 2 Diabetes Mellitus in a Segment of Indian Urban Population Rajyalakshmi Amancherla and Pradeep Naik

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Abstract No: 10.21: Association of ApoA5 1131 T>C Polymorphism with Triglyceride Levels in Indians R.Lakshmy, H.P.S Sachdev, Meenakshi Sharma, S.K Bhargava, G.R Chandak and K.S Reddy Abstract No: 10.22: Minerals Status in Thyroid Disorders (Hypo & Hyper Thyroidism) B.Suneel, D.R.Nagendra, K.Ramalingam and J.N.Naidu Abstract No: 10.23: Postprandial Hyperglecaemia and Reduction in Serum Paraoxonase Activity in Type 2 Diabetes Mellitus Subject: an Alarm of Vascular Complication Raj Narayan Gupta, Neelima Singh, S.K. Singh and Preeti Dohare Abstract No: 10.24: Anti-oxidant Potential (AOP) of LDL in Post Menopausal Women Suffering from Type 2 Diabetes Mellitus. Nivedita Singh, Neelima Singh, Sanjeev Singh and Vishal Bhargava Abstract No: 10.25: Analysis of Multivariate Factors in Metabolic Syndrome - a Correlation Study Vinayak S Bhat, Reshma Kumarchandra and Vijay S.Bhat Abstract No: 10.26: Diabetes Risk Prediction by Fructosamine in Non diabetic First Degree Relatives Hegde.A, D'souza.F, Kaveeshwar.V, Jose A, Tasneem.S and Manjrekar A.P Abstract No: 10.27: A Study of CVD Risk Factors in Metabolic Syndrome. Sajimon Thomas and T.Vijayakumar Abstract No: 10.28: CVD Risk Factors in Women with Subclinical Autoimmune Thyroiditis. Sudheesh.M and T.Vijayakumar Abstract No: 11.1: Recurrent Aphhous Ulcer and Oral Lichen Planus: a Study of Patho-physiological and Psychosomatic Significance of Salivary Nitric Oxide. AB Agte, Bm Despande, Sm Dharwadkar, Kiran Jagtap, S.B.Gaikwad and S.R.Holkar Abstract No: 11.2: Oxidative Stress and Calcium Phosphorus Ratio in Rheumatoid Arthritis Dr.A.Pullaiah and Dr.N.Vani Abstract No: 11.3: The Role of Oxidative Stress and Fibrinogen Level in Chronic in Diabetes Mellitus Mrs. G. Ramani, Dr. Rita Mary Aruna, Dr. T. V. Mohan and Mrs. G. Kavitha

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Abstract No: 11.4: The Effect of Yoga Therapy on Antioxidant Level in Perimenopausal Women Alok Kumar, Archana E., Akshatha, Mrs. Gayathry Nayak, Dr.Revathi P. Shenoy and Dr. Anjali Rao Abstract No: 11.5: Aluminum Induced Morphological and Biochemical Changes in Liver, Kidney and Brain of Male Albino Rats; An Age Dependent Study Anumesh Pathak, Sandeep Tripathi, A. K. Pathak, Abbas Ali Mahdi, Sanjay Sen, Ramkrishan Ojha and Neelam Dubey Abstract No: 11.6: Reduced Nitric Oxide Synthesis in Preeclampsia: Role of Inflammatory Cytokines and eNOS Gene Polymorphism Archana Singh., Deepika Sharma., Chitra Raghunandan and Jayashree Bhattacharjee. Abstract No: 11.7: Liver Function and Total Antioxidant Capacity in Hypothyroidism Bhawna Bhimte, Dr.B.K.Agrawal, Dr.V.K.Sharma and Prashant Nigam Abstract No: 11.8: A Study on Free Radical Activity in Burns Dr. Bikash Kr. Chaudhury and Dr. Runi Devi Abstract No: 11.9: Oxidative Stress in N-acetylcysteine Treated Off Pump CABG Surgery Patient Shyama Subramaniam, S.Subramaniam, K.Dhanajayan, K.Ezhilarasan, K.Baskar and B.Jalakandan Abstract No: 11.10: Estimation of Lipid Profile, Vitamin E and Malondialdehyde, in Pre-eclampsia Dr. Chandan Kumar Nath and Dr. Upasana Baruah Abstract No: 11.11: Antioxidant Status in Goats During Pregnancy Cynthia Jose and K.K. Jayavardhanan Abstract No: 11.12: Comparative Study of Lipid Peroxidation and Non-enzymatic Antioxidant Vitamin E and Vitamin C in Plasmodium Vivax and Falciparum Malaria in Western Rajasthan with special reference to Bikaner District. R.K. Vyas, D. Yadav, M.L. Sharma, Y. Soni and D. Nandini Abstract No: 11.13: Comparative Study of Serum Myeloperoxidase Level and Lipid Profile in Healthy Individuals Suchetha Kumari. N and Ramitha K Abstract No: 11.14: Nitric Oxide and Lipid Profile in Type 2 Diabetes Mellitus-- A Case Control Study in East Sikkim Dr. A. Ghosh, Ms. Y. Bhutia, Dr. T. A. Singh and Dr. M. L. Sherpa

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Abstract No: 11.15: MDA, Antioxidant Enzymes, their Correlation in Normotensive and Pre-eclamptic Maternal and Cord Blood Mohd Suhail, Safia Suhail, Bharat Kumar Gupta and Vinay Bharat Abstract No: 11.16: Superoxide Dismutase Activity in RidleyJopling Leprosy Types CVB Prasad and MV Kodliwadmath Abstract No: 11.17: Effect of Wheat Grass on Oxidative Stress in High Fat Diet induced Hyperlipidemia in Rabbits Dr. Kiran Dahiya, Dr. Veena Singh, Dr. Sangeeta B.Singh, Dr. Jyoti Sethi, Dr. Mridul Yadav and Mr. Prashanta Saha Roy. Abstract No: 11.18: Study of Lipid Peroxidation and Anti Oxidant Status in Pre-Eclampsia Dr. Ranjini.S, Dr. S.S. Halyal and Dr. Jayaprakash Murthy D.S Abstract No: 11.19: Attenuation of Inflammation and Oxidative Stress by Plumbago capensis in Freund's Complete AdjuvantInduced Arthritis in Rats A.I.Charles Dorni , Hannah R. Vasanthi, K.S Jayachandran and G.V.Rajamanickam Abstract No: 11.20: Effect of Vitamin E Supplement on Blood Anti-oxidant Status in Cigarette Smokers. Juhi Aggarwal and Sadhana Sharma Abstract No: 11.21: Oxidative Stress and Antioxidant Therapy in Nephrotic Syndrome Jyoti Dwivedi and Dr. Purnima Dey Sarkar Abstract No: 11.22: Antioxidant Status in Patients with Uncomplicated Type 2 Diabetes Mellitus K. Sowmya, S.Thanikachalam and Satyajeet Giri Abstract No: 11.23: Chronomics of Oxidants, Anti-oxidant Enzymes and Related Molecules in Gyenecological Malignancies R.K Singh, S.Singh, R.Singh, S.Pandey, S.Mehrotra, U.Singh, O.Schwartzkopff, G. Cornelissen and F.Halberg Abstract No: 11.24: Seminal Malondialdehyde, Nitric Oxide, Zinc and Superoxide Dismutase Activity in Male Infertility Kavita More, Prof. P.G Samant and Dr. Z.G Badade Abstract No: 11.25: Study of Nitrate + Nitrite and Antioxidant Enzyme Levels in Oral Cancer Kinjal R. Patel, Jayendra B. Patel, Shruti R. Patel, Shilin N. Shukla and Prabhudas S. Patel

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Abstract No: 11.26: Oxidatant and Antioxidant Defence Mechanism in Chronic Myeloid Leukemia Rizwan Ahmad, Anil K Tripathi, Payal Tripathi, Vinod Kumar Singh and Raj K Singh Abstract No: 12.1: Antioxidant Markers and Liver Function Parameters in Chronic Alcoholics: A Comparative Study between Koraga Tribal Community, Civilized Alcoholics and Healthy Controls Naureen Anwar, Prasiddha Tilak, Mahesh.S and Dr. Mungli Prakash Abstract No: 12.2: Oxidative Stress in Organophosphorous Toxicity Dr.M.Jaiprakash Babu Abstract No: 12.3: Effect of Lemon Grass Oil Mouthwash on Patients with Gingivitis Meghana Bhandarkar, Mahima .B.S, Madhur Agrawal, Sonal Sukreet, Dr. Shobha.U.Kamath and Dr.Meena Anand Abstract No: 12.4: Oxidative Stress and Vitamin Antioxidant Status in Rheumatoid Arthritis. Dr P.B Desai, Dr. Manjunath.S, Dr. Shivraj Gowda, Dr. Sumangala Kadi, Dr. Shivprasad and Dr. Chetana.K. Abstract No: 12.5: Changes in Advanced Oxidation Protein Product (AOPP) in Moderate and Severly Uremic Rats Merin Iype C, Subramanya Upadhya, Sharmila Upadhya and Gopalakrishna Bhat Abstract No: 12.6: Oxidative Stress and Calcium Levels in Senile and Diabetic Catract Patients Deepa K, Sumana Kamath, Vivian D'Souza and Nandini.M Abstract No: 12.7: Oxidative Stress in Erythrocytes and Plasma of Healthy Controls Nirjala Laxmi Madhikarmi and Kora Rudraiah Siddalinga Murthy Abstract No: 12.8: Oxidant and Antioxidant status in Metabolic Syndrome PVLN Srinivasa Rao, N. Manasa Kumari, Alok Sachan, KVN Raju, M.M.Suchitra, V. Seshadri Reddy and Aparna R.Bitla Abstract No: 12.9: Study of Antioxidant Enzymes of Seminal Plasma in Association with Mental Stress Subjects R.K Padalkar, V.R Pandhare, P.S Kamble and P.D Zande Abstract No: 12.10: Role of Mucuna Pruriens on Antioxidant Enzymes of Seminal Plasma in Association of Mental Stress Subjects VR Pandhare, RK Padalkar, PS Kamble and PD Zende

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Abstract No: 12.11: Relationship between ROS and Antioxidants in Male Infertility Parineeta Samant, Prof P G Samant and Dr Z G Badade Abstract No: 12.12: Study of Lipid-Peroxidation and Antioxidant Status in Malaria Patients Pawan Kare, B.K.Agrawal and V.K.Sharma Abstract No: 12.13: A Study of Oxidative Stress in Cancer Patient Before and After Supplementation of Vitamin C and E Dr. Pranami Bordoloi, Dr. Bikash Kr. Chaudhury, Dr. Chandan Kr. Nath and Dr. Dipali Das Abstract No: 12.14: Biochemical Antioxidative Parameters in Type 2 Diabetic Patients with Cardiovascular Complications. Prashant Nigam, B. K. Agrawal,V. K. Sharma and Bhawna Bhimte Abstract No: 12.15: Evaluation of Flow Rate, pH, Buffering Capacity, Calcium, Total Protein and Total Antioxidant Levels of Saliva in Caries Free and Caries-active Children ­ an in vivo Study Dr. Preethi.B.P, Dr. Jayaprakash Murthy.D.S and Dr. Reshma Dodawad Abstract No: 12.16: Comparative Study of Free Radical Activity of Plasmodium Falciparum and Plasmodium Vivax malaria Patients PS Kamble, RK Padalkar, SR Kharade, PD Zende and VR Pandhare Abstract No: 12.17: Alterations in Oxidant and Antioxidant Status in Early Alcoholic Liver Disease Ms. Sripradha. R, Dr. M.G. Sridhar and P.Doureradjou Abstract No: 12.18: Respiratory Burst Enzymes of Leukocytes in Withaferin Treated Carcinoma of Uterine Cervix Reshma K, AV Rao, Dinesh M, and Vasudevan D.M Abstract No: 12.19: Advanced Oxidative Products of Proteins and Total Antioxidants in Gastric Carcinoma Patients Anita, Sudha K, Beena V Shetty and Gayatri M Rao Abstract No: 12.20: Protein Oxidation and Antioxidants in Oral Cancer Sudha K, Vinodchandran, Gayathri M Rao and Beena Shetty Abstract No: 12.21: Oxidative Stress in Maternal and Cord Blood of Pre Term Infants Revathi.R, Saravanan.A, Ramakrishnan.T, Bharathy.N and Sujitha.S

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Abstract No: 12.22: Effect of Renoprotective Therapy on Plasma Thiols and Ferric Reducing Antioxidant Power in Children with Nephrotic Syndrome Rimshida Malik, Priya Pai, Dr. Krishnananda Prabhu, Dr. Nalini Bhaskaranand and Dr. Anjali Rao Abstract No: 12.23: Role of Lipid Peroxidation and Enzymatic Antioxidant Status in Complicated Pregnancies. Sadanand.B.Patil, M.V.Kodliwadmath and Sheela M. Kodliwadmath Abstract No: 12.24: Study of Antioxidant Status with Respect to AF after Cardiac Surgery Shalini Nair, Khalid Iqbal, Madhavi S Phadke and Jagdish Khandeparkar Abstract No: 12.25: Chronic Alcohol Induced Tissue Injury: Microarray Analyses of Gene Regulation in the Disease Process R. Sandeep, P.Nagababu, Sunitha T, Rekha.D, Farid.M., Malathi. R Ranjeetha.N and R.Polavarapu Abstract No: 12.26: Involvement of Cellular Oxidants in Deregulated Redox Homeostasis in Diabetes Mellitus P. P. Singh and Farzana Mahdi Abstract No: 13.1: Oxidative Stress and Inflammatory Response in Pre-eclampsia Sathish B.M , Bobby.Z and Habeebullah.S Abstract No: 13.2: Antioxidant Activity of Fermented Tea (Kombucha) U. Satyanarayana and T. Srihari. Abstract No: 13.3: Health Benefits of Origanum vulgare (a Medicinal Herb) through its Antioxidant Property. T. Srihari, U. Satyanarayana and N. Nalini Abstract No: 13.4: Status of Enzymatic and Non-enzymatic Antioxidant Systems in Patients with Acute Ischemic Stroke J. Sudha Rani Abstract No: 13.5: Oxidative Protein Damage in Thyroid Dysfunction Dr. Sweta Shivshanker and Dr. Anushre Prasad Abstract No: 13.6: Auto Antibodies against Oxidized Low Density Lipoproteins and Lipid Peroxidation in Patients with Essential Hypertension K.Ramalingam, K.Santha, S.Sethupathy and R.Vinoth Kumar

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Abstract No: 13.7: Oxidative Stress and Insulin Resistance in Type2 Diabetes Mellitus R.Viswakumar, K.Ramalingam and R.Vinoth Kumar Abstract No: 13.8: Lipid Profile and Glucose Tolerance Test in Cholelithiasis R.N. Devaki and H.S Virupaksha Abstract No: 13.9: Association of Hypoalbuminemia with Size of Pleural Effusion in Pneumonia. Mrs Ranjita V. Gaur and Dr (Mrs ) Rita M. Shah Abstract No: 13.10: Biochemical Effects of Feeding Diglycerol Disulphide in Rats Fed Ethanol or High Fat Diet Blessy M. I., Vinod P., Praveen K. V., Sheeba V., J. K. Mukkadan and P. K. Joseph Abstract No: 13.11: The Prevalence of Celiac Disease in Patients Presenting with Gastro-intestinal Symptoms Satinder Kaur, APS Narang, Navpreet Kaur and Indu Verma Abstract No: 13.12: Jun N-Terminal Kinase (JNK) mediates Dextran Sodium Sulfate-Induced Tight Junction Barrier Dysfunction in Caco-2 Cell Monolayers. Geetha Samak and Radhakrishna Rao Abstract No: 13.13: Storage Stability Characteristics of Hepatobiliary Enzymes in Cattle and Buffalo Divya P.D. and K.K. Jayavardhanan Abstract No: 13.14: IL-6 is a Reliable Independent Biochemical Marker for Predicitng Severity of Acute Pancreatitis G.Vasanthalakshmi, Pradeep Naik B., Prem Sagar, B. Vijay Kumar and K.Ravindranath Abstract No: 13.15: Peritoneal Fluid and Bblood Lactic Acid Levels in Acute Abdominal Disorders A.P.S. Narang, S Goyal, Indu Verma, Satinder Kaur and J.S. Multani Abstract No: 13.16: Estimation of Serum Enzymes in Antibiotic Therapy for Assessment of Hepatotoxicity Sen S, Sinha S, Nazmi A , Tripathi S and Prashant Abstract No: 14.1: Phytochemical Investigation and Anthelmintic Activity of Extracts of Seeds of Elletaria cardamomum A.A Bidkar, M. K. Aswar, K.N. Gujar and C. S. Patel Abstract No: 14.2: Phytochemical Investigation and Pharmacological Evaluation of Extracts of Leaves of Calotropis procera Linn. A.A Bidkar, K.N. Gujar, U.S. Bagul K.S.Kanugo and K.J. Oza

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Abstract No: 14.3: Phytochemical Investigations and Anticonvulsant Activity of Seeds of Brassica campestris. A. Bidkar, K. N. Gujar, P. S. Parmar, A. A. Deshpande and A.V. Vaidya Abstract No: 14.4: Anticarcinogenic Effect of Indigofera Aspalathoides by Influencing Xenobiotic Enzyme Activity During DMBA Induced Hamster Buccal Pouch Carcinogenesis Philips Abraham, V. Arul and Dr.S.Sethupathy Abstract No: 14.5: Biochemical Changes of Feeding Nigella sativa to Rats Fed a High Fat Diet and Treated with Adrenaline Bini Balakrishnan, Ceema Varghese, Nithi Varghese, Vidya A B, Praveen K.V, Sheeba Varghese, J. K Mukkadan and P.K Joseph Abstract No: 14.6: Biochemical Changes of Feeding Desmodium gangeticum to Rats Fed a High Fat Diet and Treated with Adrenaline Ceema Varghese, Bini Balakrishnan, Nithi Varghese, Vidya A B, Praveen K.V, Sheeba Varghese, J.K Mukkadan and P.K Joseph Abstract No: 14.7: Comparative Study of Jamun Powder and Tulsi Leaf Powder as Hypoglycemic and Hypolipoidemic Agent in Diabetes Mellitus Type 2 Dr. Meena Verma, Deependra Sharma, Dr. Sangeeta Paneri and Dr.A.Mishra Abstract No: 14.8: Nyctanthes arbortristis Extract Restores the Cartilage Damage During Experimental Arthritis Brijesh Rathore, A .A Mahdi, B.N Paul, P. N Saxena, Farzana Mahdi and S. K Das Abstract No: 14.9: Polyherbal Anti Diabetic Drug: an Approach to Cure Diabetes Gaikwad S.B, Bhagwat V.R and Trivedi D.J Abstract No: 14.10: Extraction and in vitro Evaluation of Biological Activity of Crude Polyphenolic Extracts from Indian Grapes (Vitis vinifera) on Transformed (AW13516) and Non-transformed (HaCaT) Stratified Epithelial Cells. Neha Tamhankar, Geeta Ibrahim and Milind Vaidya Abstract No: 14.11: Beneficial Effects of Tetrahydrocurcumin and Chlorogenic Acid on Lipid Profile in Streptozotocin-NicotinamideInduced Diabetic Rats Karthikesan.K and Pari.L Abstract No: 14.12: Search for a Perfect Diet Including Antioxidants Prof (Dr) Kaushal K.Srivastava

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Abstract No: 14.13: Antioxidant Evaluation of Some Hepatoprotective Medicinal Plants Chorge Ketan, Dr. (Mrs.) Kale K.U and Chorge Ulka Abstract No: 14.14: Does Chilli Affect Platelet Aggregation? Kiran DK Ahuja, Murray J Adams, Dominic P Geraghty and Madeleine J.Ball Abstract No: 14.15: Antidiabetic Effects of the Aqueous Extract of Fruit of Withania Coagulans in Diabetic Albino Rats Kirtikar Shukla, J.K Gambhir and Rimi Shukla Abstract No: 14.16: A Putative Anticancer Mechanism of Plant Derived Polyphenols Mohd. Fahad Ullah, Uzma Shamim, Sarmad Hanif, Asfar S Azmi, Showket H Bhat and S.M.Hadi Abstract No: 14.17: A Comparative Study of Nitric Oxide and Hydrogen Peroxide Scavenging Activities of Botanical Extracts Ocimum sanctum and Wagatea spicata. Geetha Samak, Revathi P Shenoy and D. M. Vasudevan Abstract No: 14.18: Cardioprotective Effect of Terminalia arjuna on Isoproterenol Induced Myocardial Infarction in Rats S.K. Shukla, S. Dwivedi, U.R. Singh, and S.B. Sharma Abstract No: 14.19: Aluminum Induced Changes in Blood and Brain; Protective Role of Bacopa monnieri Sandeep Tripathi, Abbas Ali Mahdi, Mahdi Hasan, Deepa Prajapati, Sanjay Annarao, Raja Roy and C. L. Khetrapal Abstract No: 14.20: Hypolipidemic Activity of Hibiscus rosa sinensis (root) in Hyperlipidemic Rats Vishnu Kumar , Ranjana Singh , Sushma Singh , Shivani Pandey , Ashok Kumar Khanna , Ramesh Chander ,Pradyumn Singh, Farzana Mahdi , Jitendra Kumar Saxena , Vinod Kumar Singh and Raj Kumar Singh Abstract No: 14.21: Cholinesterase Activity in Health Workers Involved in Handling and Spraying of Organophosphorous Insecticides Himanshu Madaan, Veena Singh Ghalaut and Ashuma Sachdeva Abstract No: 14.22: Lead Poisoning in Residential Areas Near Drainage Systems in Bangalore Anitha Jerry Juneeja Varghese and Antony.P.U Abstract No: 14.23: Study of ADA Levels in Tuberculus and Non tuberculus Pleural Effusion K. Sravanthi, Dr. B. Prabhakar Rao, Dr. V. Sunanda and Dr. C.N. Prasad

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Abstract No: 14.24: A Brief Study of Lead Poisoning Among Children Faculty and Students Abstract No.14.25: Iodine Deficiency Disorders Among the Primary School Children of Eastern Nepal. Gelal.B, Chaudhari.R, Nepal.A, Das B.K.L, Lamsal.M and Baral.N Abstract No.14.26: Role of Plasma Homocysteine Level in Coronary Artery Disease and Stroke- A Clinical Perspective Girish P. Pandey, Sukhes Mukherjee, Kannan Vaidyanathan and D.M Vasudevan Abstract No.14.27: Vitamin B12 and Folic Acid Levels in Tobacco Chewers: A Duration Dependent Study. Indira Samal, M.Maneesh, Sukhes Mukherjee and Subir K Das . Abstract No.14.28: Role of Pro and Anti-inflammatory Cytokines in the Modulation of alcoholic Liver Disease. Sukhes Mukherjee, Subir Kumar Das, and D.M Vasudevan Abstract No.14.29: Urine glucose estimation Imbichi Mammi Abstract No.15.1: Antidiabetic Activity of a Coastal Mangrove, Rhizophora mucronata Poir. : Effect on Carbohydrate Metabolism in Alloxan-induced Diabetic Rats K. Desigamani b , S. Manivannan * , K Kathiresan a and T. Ramanathana rita mary arunab Abstract No.15.2: Antitumour Activity of Some Homoeopathic Medicines in Animal Models and Possible Mechanism of Action Ramadasan Kuttan, Preethi K.C., Girija Kuttan Abstract No.15.3: Radioprotective and Chemoprotective Effect of Carotenoid Meso-zeaxanthin A.P. Firdous and R. Kuttan Abstract No.15.4 Hepatoprotective Activity of Carotenoid Lutein Sindhu E R and Ramadasan Kuttan Abstract No. 15.5: XRCC ­ 1 Polymorphisms (Arg194Trp and Arg399Gln) in Oral Cancer from a Kerala Population. Balaji Natarajan Abstract No.15.6: Multiple Myeloma, 2008 Statistics Ramesh K, Riju M, Prasanth G, Sreejith A.S.,

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CONTENTS

Abstract No. 15.7: Auto Immune Thyroid Disease Antibodies Riju M, Prasanth G, Sreejith A S. Ramesh K, Abstract No.15.8: Orocecal Transit Time and Small Intestinal Bacterial Overgrowth in Patients with Microscopic Colitis SV Rana, KK Prasad, SK Sinha, S Sharma, J Kaur and K Singh. Abstract No.15.9: Plasma Myeloperoxidase in Diabetes Mellitus Usha Anand, Vijaya D, Gayathri B, Aruna V and Anand CV Abstract No.15.10: Serum Cystatin C in Various Stages of Chronic Kidney Disease Krishnamurthy N 1, Usha Anand 2, Anand C.V 2, Aruna V 2 and Venu G 3 Abstract No.15.11: Evaluation of Hyperglycemia, Glucose Intolerance, Hypertension and Socioeconomic Position in Eastern Nepal Mehta KD1, Karki P2, Lamsal M1, Paudel IS3, Majhi S1, Das BKL1 Sharma S2 Jha N3 and *Baral N1 Abstract No. 15.12: Advanced Research on Heavy Metal Pollution in Ganga Water; Detection with Integrated Coupled Plasma-Atomic Emission Spectrometry Ashutosh Kumar Sharma Abstract No.15.13: My Experience of Two Decades Using Mid Size Laboratory Automation in Clinical Laboratory Practice Dr. Ulhas R. Tendulkar, Director, Ambalab, Goregaon East, Mumbai-400063 Abstract No. 15.14: Detection of Mitochondrial Point Mutation in Maternal Inherited Diabetes with Deafness (MIDD) Using Various Molecular Tools R.R. Kumar, K. Ponsuganthi, S. Selvaraj*, H. Palaniswamy#, B. Mohanty Abstract No. 15.15: Gold in Human Semen Around and Away from a Gold Deposit Area Sahab Khan P, Skandhan K Pa, Ajesh K, Siraj MVP. Abstract No. 15.16: Effect of Chilli (Capsaicin)on Oxidation of Serum Lipoproteins Kiran D.K Ahuja and Madeleine J. Ball

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ORATIONS

O-1.

Dr.T.N Pattabhiraman Oration-Laboratory Overload: How to Optimize?

Brig MM Arora

Professor & Head, Dept of Biochemistry, Armed Forces Medical College, Pune-411040. Email: [email protected]; Lt. Col. (Mrs) J.k Bhatia, MD, Classified specialist pathology, Military Hospital, Dehradun (UP) Background: Medical fraternity requisitions diagnostic tests for multiple reasons. More often than not, the tests lead to more tests either to exclude or to confirm doubts raised by the test results. These tests do have inherent morbidity, discomfort and cost. Growing expenditure on diagnostic tests without matching improvement in the health status warrants an internal audit of the laboratory utilization. Methods: A retrospective utility audit was done for certain routinely advised laboratory tests. Tests included in audit were blood urea estimation in annual as well as periodic medical examination; bleeding and coagulation time in pre-anesthetic check-up; and aspartate aminotransferase (AST); antibodies to hepatitis C virus (anti-HCV) in diagnostic work-up of acute onset jaundice; and T3, T4, TSH estimation in evaluation of thyroid function. acute onset jaundice, 6049 AST estimations in 1024 patients did not contribute anything more than what was inferred by alanine aminotransferase (ALT). Prevalence of anti HCV antibodies in acute onset jaundice in serving soldiers (11 out of 1225; 0.89%) though more than that in the blood donors from the same population (17 out of 4105; 0.41%) was less than anticipated false positives (18 out of 1225; 1.5%) as per the claimed specificity (98.5%) of the test kit. None of the 2766 bleeding and clotting time tests detected a bleeding or coagulation disorder. Despite sizable evidence in favour of TSH being an adequate single screening test for evaluation of thyroid, the ordering frequency of T3, T4, and TSH was nearly the same.

Conclusion: The study reveals gross overuse of the laboratory that may not be good for the patient and the organization in terms of direct costs as well as indirect costs due to false positive results. To make the matters worse, this laboratory overload Results: During the period of study, 793 adversely affects the quality and availability of individuals underwent AME / PME. In none of laboratory results. Therefore, a test should only them urea estimation provided any additional be advised, if positive or negative result would information which was not inferred by serum dictate a change in patient management. creatinine. Similarly, in diagnostic workup of

O-2.

K.E.M. Hospital and Seth G.S. Medical College Oration Regenerative Nanomedicine - Prospects for Diagnostics and Therapy using Nanomaterials

Shanti Kumar Nair

Dean of Research, Head of Nanotechnology Division, Amrita Institute of Medical Colllege, Kochi. Nanotechnology has shown great potential as a for augmenting the body's healing capacities in primary diagnostic and therapeutic tool in the the area of regenerative medicine through novel medical sciences. It has also shown its potential approaches such as tissue or organ engineering.

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The diagnostic potential of nanotechnology has been advanced by the development of novel nanoparticles with luminescence or superparamagnetic or scintillator properties. Such nanoparticles can be tagged to diseased tissues, such as cancer cells and detected by their relevant scale-modulated property. Tagging of nanoparticles to cells has emerged as a new science involving interaction of chemical moieties with the nanoparticle surface and with the cell membrane or with monoclonal antibodies. Successful tagging of nanoparticles has opened the door to cell-specific rather than systemic therapy. In cell-specific therapy the drugs are delivered specifically to the target cells by

conjugating the drugs with the nanoparticles or encapsulating the drugs within nanoparticles. Other therapeutic approaches involve photodynamic or thermal therapies which do not involve highly toxic chemotherapeutic agents. In regenerative medicine also nanotechnology has shown the potential to enhance the regenerative ability by its ability to control cell-material interactions. The control of cell-material interactions using nanomaterials has emerged as a new and dynamic science with great potential for healing and regeneration of tissues and organs. In this presentation the author will present some of the data from his own lab as well as some data from other leading scientists in the area.

O-3.

Mrs. & Dr.G.P Talwar Oration Endothelial Dysfunction with Special Reference to eNOS Gene Polymorphism: Another Plausible Mechanism for CAD Risk in Postmenopausal Women

Jayashree Bhattacharjee Department of Biochemistry, Lady Hardinge Medical College, New Delhi, India

Background: Framingham study indicates more than two fold age related increased risk of Coronary artery disease (CAD) in postmenopausal women as compared to premenopausal women. Nitric Oxide (NO) is an important regulator of endothelial functions and has several cardio protective actions as vasodilation, inhibition of platelet aggregation & adhesion. The Glu298Asp polymorphism of endothelial nitric oxide synthase gene (eNOS) is associated with CAD risk. Estrogen exerts its local vasodilating and antiplatelet actions as a consequence of endothelial response mediated by release of EDRF (endothelial dependent relaxing factor), which is NO. The purpose of this study was to assess CAD risk in postmenopausal women by evaluating Nitric oxide ­ Cyclic GMP signal transduction pathway & screening Glu298Asp polymorphism along with other established CAD risk markers for the purpose of early diagnosis and intervention. Methods: Two consecutive studies were conducted. Each study design consists of 50 postmenopausal women cases compared with 50 premenopausal women controls for similar age group excluded for any known CAD risk. In the first study we measured the levels of NO, Estrogen, Apo-B and functional relation of NO and cGMP to evaluate signal transduction pathway. The second study was done to investigate the associated role of Glu-Asp298 eNOS gene polymorphism & platelet function in increasing postmenopausal CAD risk. Results: Plasma NO, Estrogen and HDL levels was found to be significantly lower (P<0.05) while VLDL and Apo-B levels shown significant increase (p<0.05) in postmenopausal women cases

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as compared to Premenopausal women controls. Plasma cGMP levels were significantly lower (P<0.05) in cases than controls. A significant positive correlation was observed between NO and estrogen (P<0.01) whereas negative correlation was found between NO and PF4 (P<0.01). The GG genotype was found in 41 subjects (82%) of the study group and 45 subjects (90%) of the control group while GT genotype was seen in 9 subjects (18%) of study group and 5 subjects (10%) of control group. No TT genotype was found. The significant decrease was observed in NO levels of GG+GT, GG and GT genotypes of study group as compared to control group.

Conclusion: the low no and low cgmp activity found in our study reflects defect in no-cgmp signal transduction which leads to endothelial dysfunction. as cad is a multifactorial disease, other factors might also play a major role in endothelial dysfunction to increase cad risk in postmenopausal women. observations found in our study suggest that enos glu298asp gene polymorphism and nitric oxide levels may be associated with an increased risk of coronary artery disease in postmenopausal women. further studies are needed to explore more mechanistic basis of the association which in turn will modify therapeutic and diagnostic strategies.

O-4.

K.L Gupta Memorial Oration Tyrosine Kinases from Basic Science to Therapy

T.S.Ganesan

Chairman, Cancer Institute & Institute of Molecular Medicine,Amrita Institute of Medical Sciences, AIMS P.O. Kochi, Kerala, 682 041 Protein phosphorylation is a central event in cellular physiology. Phosphorylation on tyrosine residues by tyrosine kinases is particularly important as it is subverted in malignant transformation. This is apparent with the discovery of a number of oncogenes which are tyrosine kinases. It is now clear the number of families of tyrosine kinases in the human genome and is restricted to metazoans. Research into the role of tyrosine kinases in malignant transformation has uncovered many new targets for treatment. The first such kinase is the `abl' kinase, but more kinases are now druggable. This whole area of research represents an excellent examples of how basic research has led to discovery of new treatments for cancer.

O-5.

Awadesh Saran Memorial Oration Lead Poisoning: Current Status in Developing Countries and the Global Perspective.

Thuppil Venkatesh

Professor Emeritus. St John's Medical College Bangalore and Principal Advisor Quality Council of India, Delhi. Lead being one of the toxic heavy metals globally considered as the number one environmental and

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health hazard affecting people of all age groups has maximum effect during growth and development of children. With its deleterious effects on all organs in the human body, lead poisoning is widely recognized as a major public health problem all over the world. Children and women are found most vulnerable to lead poisoning especially in developing countries. Unlike other health hazards, lead poisoning is hundred per cent preventable when legislation is in place. The estimate by the World Health Organization has indicated that over 120 million people are overexposed to lead all over the world and 99 percent of the most serious cases are in the developing world. The project Lead Free of The George Foundation was initiated in Bangalore and its vicinity, and subsequently expanded to several other major cities across India), found that more than 51.3 per cent of children in Indian metros below 12 years of age have their blood lead levels above 10 ug/dl. Blood lead levels above 10ug/dl is known to reduce the IQ during growth and developmental phase of children. An average reduction of IQ to an extent of 4-6 units on a scale is known to result in the economic damage to countries to the tune of over Rs 600 crore annually by 2015. Author was involved in the introduction of unleaded gasoline in India in March 2000, and has recently found out other major sources such as lead based traditional medicines and lead-based paints. Detailed studies by the author has indicated that though the per capita consumption of paints in India is around one-fifteenth of developed

countries with a demand as on date with the unprecedented boom in the housing sector which is expected to satisfy the demand for over 30 million new homes which is a great threat to the countries future economy. A study report by the author during 2006 on the analysis of lead in paints in China, India and Malaysia revealed that over 80 new paints from these countries used in homes had very high lead levels -- in fact, exceeding the levels of 1000 ppm. Though there is a need for the ban of lead-based paints all over the world, which has been emphasized time and again to prevent exposure and associated sicknesses, developing countries have not taken this seriously. In India, for example, lead-based paints were found in the homes of three children whose blood lead levels were at least 40 ug/dl. In a report on dust lead levels in Delhi, homes had dust with lead levels of 31 per cent. That incidentally exceeds the current US limit of 40 ug/sq ft. Apart from this almost 80% of all the lead now used in many parts of the world is for the production of lead-acid batteries and that in India and other developing countries the environmental friendly recycled lead is of much lower proportion in lead acid battery manufacturing causing another major threat to both environment and health. Author is involved in the development of the BEST certification program to all lead acid battery industry in the country with wide acceptability. With the recent efforts to create lead map of two megacities in India the National Referral Center for Lead Poisoning in India established by the author with several branches all over the country is conducting lead educator program sponsored by QCI is working towards lead safe society.

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KEY NOTE ADDRESSES KN-1.

Vitamin E is activated by phosphorylation to ?-tocopheryl phosphate

Angelo Azzi

Vascular Biology Laboratory, JM USDA-HNRCA at Tufts University, Boston, MA 02111, USA. Vitamin E (-Tocopherol), traditionally described as an antioxidant, has revealed to posses specific cellular functions that are independent of its radical scavenging properties. It inhibits protein kinase C and PI3 Kinase, as well as activates protein phosphatase 2A and diacylglycerol kinase. -Tocopherol - Kinase Phosphatase the effects caused by á-TP to cells, in a dose-dependent way. In a search for genes that are specifically and strongly affected by TP, gene arrays representing essentially all human genes were used. A group of genes was found that is up-regulated by TP but not by T; suggesting that in some cells T is not sufficiently converted to TP. Genes which were regulated by both compound were more affected by TP than by T. References 1. Azzi, A. (2007) Molecular mechanism of alphatocopherol action, Free Radic Biol Med. 43, 1621. 2. Gianello, R., Libinaki, R., Azzi, A., Gavin, P. D., Negis, Y., Zingg, J. M., Holt, P., Keah, H. H., Griffey, A., Smallridge, A., West, S. M. & Ogru, E. (2005) Alpha-tocopheryl phosphate: a novel, natural form of vitamin E, Free Radic Biol Med. 39, 970-6.

-Tocopheryl phosphate -

-Tocopheryl phosphate receptor -

SIGNALING GENE EXPRESSON 3. Negis, Y., Zingg, J.-M., Ogru, E., Gianello, R., CELL MODULATION Libinaki, R. & Azzi, A. (2005) On the Existence of Cellular Tocopheryl Phosphate, its Synthesis, Furthermore, at transcriptional level, several genes Degradation and Cellular Roles: A Hypothesis (the first to be described have been CD36,TTP,-tropomyosin, and collagenase) are moduIUBMB Life. 57, 23 - 25. lated by -tocopherol [1]. -Tocopheryl phosphate (á-TP) is synthesized and hydrolyzed in animal 4. Munteanu, A., Zingg, J.-M., Negis, Y., Azzi, A., Ogru, E., West, S., Libinaki, R. & Gianello, R. cells and tissues; it modulates also several cell (2004) Modulation of cell proliferation and gene functions [2, 3]. While it is similar to á-tocopherol expression by ?-tocopheryl phosphates: (á-T), á-TP appears to be more potent than á-T in Relevance to atherosclerosis and inflammation, inhibiting cell proliferation, down regulating Biochemical and Biophysical Research CD36 transcription, inhibiting atherosclerotic Communications. 318, 311-316. plaque formation etc. In cells and animals á-TP does not act by liberating á -T; rather, the intactmolecule appears to be more potent than á- 5. Negis, Y., Aytan, N., Ozer, N., Ogru, E., Libinaki, R., Gianello, R., Azzi, A. & Zingg, J. T itself [4, 5]). á-TP can be defined as the active M. (2006) The effect of tocopheryl phosphates form of á-T. Administration of á-TP to cells or to on atherosclerosis progression in rabbits fed animals requires its transfer through membranes. with a high cholesterol diet, Arch Biochem Specific inhibitors, glybenclamide and probenecid Biophys. 450, 63-6. showed to inhibit á-TP transport and to eliminate

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KN-2.

Impact of Pharmacogenetics on Therapeutic Drug Monitoring

M. Oellerich, V.W. Armstrong and N.Von Ahsen Department of Clinical Chemistry, George-August-University Göttingen, Germany Pharmacogenetics and its application to TDM is an emerging field. Gene test guided therapy could facilitate the selection of a drug and its dosage to which the patient best responds. A future goal may be a pharmacogenetic patient card. Valid pharmacogenomic biomarkers listed by the FDA include drug metabolising enzymes (CYP2C9+, CYP2C19, CYP2D6, DPD, NAT, TPMT + , UGT1A1 + ) as well as HLA-B*5701 + and VKORC1+. Genomic biomarkers for which testing is recommended are marked (+). There is a lack of evidence supporting a clear association between ABCB 1 (MDR1) genotype and clinical drug response or toxicity. Variability of P-glycoprotein expression can affect intralymphocyte concentrations of cyclosporine. It could be demonstrated that the intracellular T-lymphocyte cyclosporine concentration has a potential for predicting rejection. The CYP2D6 polymorphism plays an important role in the metabolism of antipsychotic as well as antidepressant drugs and tamoxifen. Due to multiple copies of a functional allele in ultrarapid metabolisers, a standard dose of antipsychotic or antidepressant drugs leads to low plasma concentrations and a lack of therapeutic effect. Ultrarapid metabolisers are predominantly found in Saudi Arabia and Ethiopia (allele frequency 10 ­ 29 %) and according to our own investigations in Papua New Guinea populations (12 %). Tamoxifen is used for the treatment of patients with hormone-dependent breast cancer. It is converted by CYP3A and CYP2D6 to active metabolites which are more potent than the parent drug. Time to disease progression was significantly shorter in Asian patients treated with tamoxifen who were poor metabolisers (CYP2D6*10/*10), compared with other genotypes evaluated. CYP2D6 genotyping may help to identify breast cancer patients who will benefit from tamoxifen. Thiopurine-S-methyltransferase (TPMT) deficiency causes azathioprine intolerance accompanied with severe myelosuppression. Testing for TPMT phenotype/genotype is a useful practice for a safer management of patients requiring thiopurine drugs. An inosine triphosphatase mutation (ITPA 94C>A) appears to be a further promising marker for azathioprine interolerance. A polymorphism in the CYP3A5 gene leads to deficiency of the enzyme and may in part contribute to interindividual differences in CYP3A dependent drug clearance. Recent investigations have shown a significant difference in dose-adjusted tacrolimus blood levels between CYP3A5 expressors compared to non-expressors. HLA-B*5701 genotyping is useful to identify patients at greatest risk for a hypersensitivity reaction to abacavir. Genomewide assessments of gene expression are increasingly being used and, e.g., have identified SLCO1B1 variants that are strongly associated with statin-induced myopathy. Expression profiling has the potential to identify gene-expression patterns related to drug response. Both approaches may be complementary and take into account that the response to most drugs is influenced by multiple genes. Drug disposition marker genotyping is a promising tool in addition to TDM for individualisation of drug dosing with the goal to improve the efficacy and safety of pharmacotherapy.

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INVITED LECTURES

IL-1.

The role of Bone Turnover Markers in Clinical Management of Osteoporosis

SD Vasikaran

Royal Perth Hospital, Perth, Australia The diagnosis of osteoporosis is based on bone mineral density measurement (BMD). The decision to treat is better assisted by the calculation of absolute fracture risk, which incorporates age and clinical risk factors in addition to BMD. Current algorithms for the calculation of fracture risk do not incorporate bone turnover markers since there are inadequate data for their use in risk calculation. In clinical practice bone turnover markers may help make cost effective treatment decisions in patients with borderline absolute risk. IL-2. Bone turnover markers which show an early response following commencement of treatment are useful for the monitoring of therapy for osteoporosis for which the use of BMD is increasingly being questioned. The use of bone turnover markers helps to identify non-compliance and non responders early. In addition to the above issues, this talk will address individual bone turnover markers which are of most value in clinical practice.

A New Diagnostic Paradigm: Some Enabling Technologies

Ross Barnard

School of Chemistry and Molecular Biosciences, The University of Queensland, St.Lucia Qld, Australia The traditional approach to diagnosis is one that has been practised since the time of hippocrates. It entails forming an hypothesis (a presumptive diagnosis) followed by design of specific tests to refute or confirm the presumptive diagnosis. Until recently this has been the basis for design of molecular tests for microrganisms, including viruses. A persistent problem is differential diagnosis, when presentation is similar for a range of possible infectious agents (for example, a wide range of arboviruses; H5N1 ("avian") versus H1N1 ("swine") or H3N8 ("equine") influenza). There are several forces operating to move us away from this model towards a "one test model. These are: 1) a need to quickly detect

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unknown, unanticipated or emerging pathogens, 2) the pressing need to rapidly acquire more clinically useful information about pathogens (for example drug resistance, host preference and pathogenicity "signatures" in influenza a, 3) multiplexing, that is, performance of many tests simultaneously and 4) the need for point-of-care or field testing. These forces are driving innovation in molecular methods, bioinformatics and devices. Some examples of these innovations will be given from our work on flavivirus testing, influenza a testing, virus drug resistance testing and a new, multiplexable, real time pcr technology.

Indian Journal of Clinical Biochemistry Supplement, 2009/24

IL-3.

The Natural Phenolic, Malabaricone B Selectively Kills the Cancer Cells through Mitochondrial Pathway

S. Chattopadhyay

Bio-Organic Division, Bhabha Atomic Research Centre, Mumbai ­ 400085, India. Redox dysregulation originating from metabolic alterations and dependence on mitogenic and survival signaling through reactive oxygen species represents a specific vulnerability of malignant cells that can be selectively targeted by redox chemotherapeutics. Simultaneous modulation of multiple redox sensitive targets by these agents can overcome cancer cell drug resistance, a common impediment of anti-cancer drug development. Search for suitable experimental chemotherapeutics that induce positive deviations from redox homeostasis through prooxidant action has therefore drawn much attention. A wide array of natural phenolics, such as turmeric, epigallocatechin gallate, resveratrol, as well several established drugs such as bleomycin, cisplatin, etoposide and paclitaxel exert their anti-cancer action through the prooxidant mechanism. The diarylnonanoid, malabaricone B, isolated from the Indian spice, Myristica malabarica was found to possess chemopreventive property against A549 (human lung carcinoma), A375 (human malignant melanoma), and Jurkat (human T cell leukemia) especially the A549 cells. It caused apoptotic killing IL-4. of the A549 cells via activation of caspase-9 and caspase-3, but not caspase-8 by altering the BaX/ Bcl-2 ratio. This revealed the mechanism of its action was primarily linked to mitochondrial damage, inducing the release of cytochrome c into the cytosol and apoptosome complex formation. The mal B-induced mitochondrial depolarization, subsequent cytochrome c release and cell cycle arrest were also confirmed with the A549 cells. Interestingly, the cytotoxicity of the phenol proceeded in a p53-independent manner, as revealed from our results with stably transfected A549- p53 cells. Mutation of the tumor suppressor protein p53, which serves as a pivotal component of the apoptosis pathway(s) is believed to be an important factor in poor prognosis of cancer and plays a critical role in the resistance of cancers to radiation and chemotherapy. Also, compared to the DNA targeting compounds, chemotherapies, aimed at mitochondrial inhibition can selectively kill tumour cells. In view of these, and our results with malabaricone B on the A549 cells suggested it to be a promising anti-cancer agent for further evaluation

Marine Brown Algae with Potent Antioxidant and Radical Scavenging Activities

M.S.Kanthimathi, 1Ghaitri Nadarajan and 2Khoo Kong Soo 1 Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia, 2 Department of Bioscience and Chemistry, Faculty of Engineering and Science, Tunku Abdul Rahman University, 53300, Setapak, Kuala Lumpur, Malaysia. [email protected] The ocean has always been a rich source of food and medicines for humans. Fairly recently, researchers have turned to the sea for alternative sources of medicinal drugs and supplements for health and wellness. However, procedures for the extraction of bioactive components from marine organisms vary from researcher to researcher. We tested the efficiency of various popular protocols used in different labs and estimated various antioxidant and free radical scavenging activities as

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Indian Journal of Clinical Biochemistry Supplement, 2009/24

markers for efficient extraction of antioxidants from 2 species of brown algae, Padina tetrastromatica and Turbinaria ornata. The best protocol was 72 hours sequential extraction at room temperature with hexane, followed by dichloromethane, acetone, ethyl acetate and

methanol. The activities estimated were the FRAP value, scavenging activities for the DPPH radical, the superoxide anion, and the hydoxyl ion, total phenolic content as well as the antiproliferative activity towards two breast cancer cell lines.

IL-5.

Male Infertility: Role of Oxidative Stress and Some Possible Herbal Remedies

Abbas Ali Mahdi, Mohd. Kaleem Ahmad, Kamla Kant Shukla, Singh Rajender**, Satya Narain Shakhwar* and Sohail Ahmad***

Departments of Biochemistry and *Urology, C.S.M. Medical University, Lucknow-226003, **Central Drug Research Institute, Lucknow-226001, ***State Government T.T. College (Unani Medical College), Lucknow-226003. Infertility may be defined as failure to conceive by a couple after 12 months of unprotected sexual intercourse. Infertility affects 15% of all couples and approximately 50% of these have an abnormality detectable in the male partner as the cause of infertility. Specific and directed treatment for male infertility is not available due to unexplained and heterogeneous nature of the disorders. Under such circumstances, only assisted reproductive techniques are of some help. However, these treatments are expensive and inaccessible to all. The lack of available specific therapies for men with infertility demands the exploration of alternative therapies. Given the lack of knowledge about etiological factors, a nondirected but general therapy may yield good results in a subcategory of patients. The rationale for the use of these therapies is based on the speculation that some forms of male infertility are caused by oxidative insult and hormonal imbalance and the use of alternative therapies may improve male fertility potential and semen quality. Ayurveda and Unani systems of medicine have documented several Indian herbs which have been in use for treatment of various disorders of male reproductive system. Many scientific studies have documented the beneficial effect of treatment of infertile men with these plants, supporting the

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literature regarding their medicinal usage. After an in depth search of literature, we came across more than 25 plants, the parts of which had been used in male infertility treatment. On the basis of information available, we made a priority list of the plants for scientific studies. We conducted studies in infertile men using seeds of Mucuna pruriens and root powder of Withania somnifera. Both these plants were highly beneficial in treating male infertility. The plants were particularly helpful in alleviating oxidative stress, which may be one of the reasons for their beneficial effect. W. somnifera had better anti-oxidant properties than M. pruriens. We also conducted experiments on normal and fertility compromised rats using these plant products. We could replicate the results in rats as far as improvement in spermatogenesis is concerned. The current status of scientific studies on fertility enhancing plants and the results from our own studies on male infertility treatment will be discussed in detail during the presentation. References: 1. Shukla, K.K., Mahdi, A.A., Ahmad, M.K., Shankhwar, S.N., Jaiswar, S.P., Tewari, S.C., (2007) Effect of Mucuna pruriens on stress associated biochemical changes in seminal

Indian Journal of Clinical Biochemistry Supplement, 2009/24

plasma of infertile males. e-CAM. (Available on line December 18, 2007) 2. Ahmad, M.K., Mahdi, A.A., Shukla, K.K., Jaiswar, S.P., Ahmad, S. & Islam, N. (2008) Effect of Mucuna pruriens on semen profile and biochemical parameters in seminal plasma of infertile men. Fertility and Sterility. 90(3) : 627635. 3. Shukla, K.K., Mahdi, A.A., Shankhwar, S.N. and Ahmad, M.K. (2008) Effect of Mucuna pruriens on hormonal status and semen quality in infertile males. Contraception. Vol. 78 (2): 194. 4. Hukla, K.K., Mahdi, A.A., Ahmad, M.K., Shankhwar, S.N., Singh R, Jaiswar, S.P. (2008) Mucuna pruriens improves male factor fertility by its action on hypothalamus-pituitary-gonadal

axis. Fertility and Sterility. Available online from October 28, 2008. 5. Mahdi, A.A., Shukla, K.K., Ahmad, M.K., Shankhwar, S.N., Jaiswar, S.P. and Chander. R. (2009) Withania sominifera reduces stress and improves the quality of semen in normozoospermic infertile men. e-CAM [In press]. 6. Ahmad, M.K., Mahdi, A.A., Shukla, K.K., Islam, N., Singh R., Madhukar, D., Shankhwar, S.N. and Ahmad, S. (2009) Withania somnifera improves semen quality by regulating reproductive hormone levels and oxidative stress in seminal plasma of infertile males. Fertility and Sterility (F and S6491R1) [In press].

IL-6.

[(E)-HST-1], 3'-5' Hydroxylated Congener of Resveratrol, A Better Anticancer Agent

Sandip K Bandyopadhyay

Department of Biochemistry, Institute of Post Graduate Medical Education & Research KPC Medical College & Hospital, Kolkata. Resveratrol is potentially of interest for the treatment of different cancer but its use was constrained because of its ulcerogenic adversative effects. We have already reported that 3` -5 hydroxylated congener [(E)-HST-1] of Resveratrol was devoid of ulcerogenic properties; hence in this study we compared its chemopreventive potential with Resveratrol in different cancer cell lines and analyzed the probable molecular mechanism of its action. Our results revealed that HST-1 could exert better antiproliferative effects on different cancer cell lines compared to Resveratrol but showed least toxicity to normal cells. HST-1 treatment induced apoptotic cell death as evident from externalization of phosphatidylserine, DNA fragmentation and cell cycle arrest at G0/G1 phase. In our pursuit to dissect molecular mechanism the cells were treated with caspase inhibitor which confirmed the involvement of caspase-3 and caspase-9 in HST-1 treated apoptosis.To confirm the activation of mitochondrial death signal, the change in membrane potential was analyzed in HST-1 treated cells which was further substantiated by the release of cytochrome c in cytosol from mitochondria. Our intricate investigation revealed that HST-1 treatment elicited intracellular ROS generation and pretreatment of intracellular ROS scavenger NAC gave significant protection to the cells treated with HST-1 . NAC treatment prevented the loss of mitochondrial membrane potential and also reduced the cytochrome c releases followed by inhibition of caspase activation in HST-1 treated cells. These observations thus confirmed that HST-1 a potentially better congener of Resveratrol exerted chemopreventive effects through mitochondrial death cascade which is modulated mainly by generation.

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IL-7.

Oxidative Stress Management: Therapeutic Approach for Depression

Naheed Banu, Ayesha Zafir, Shoa Naqvi and Ghazala Mehdi *, Department of Biochemistry, Faculty of Life Sciences,*Department of Pathology, J. N. Medical College, A.M. University, Aligarh, U.P., India, [email protected] Objective: Anomalies in oxidative biology are now recognized as a critical pathophysiological mechanism for stress and depression. Biochemical evidence that depressive behaviour is indeed associated with deficits in the protective antioxidant system is still lacking. Recently we showed that augmentation of in vivo antioxidant defenses might provide a convergence point for the neuroprotective effects of different antidepressants used for treating stress disorders. On the basis of these findings, we tested the hypothesis that micronutrients like folic acid and ascorbic acid could conversely induce an antidepressant-like effect in rodents. Methods: The impact of chronic treatment with folic acid, ascorbic acid or the SSRI fluoxetine was investigated on the behavioural outcomes of chronic restraint stressed rats in association with assessment of brain antioxidant status, biomarkers of oxidative damage, changes in body weight and histological lesions. Results: Restraint stress caused anhedonia and behavioural despair, along with depletion of antioxidants SOD, CAT, GST, GR, GSH, and increase in lipid peroxidation and protein carbonyl contents. As with ascorbic acid, treatment of restraint-exposed animals for 21 days with folic acid reversed the altered antioxidant defenses. Stress induced behavioural changes were attenuated by both folic and ascorbic acids, similar to fluoxetine, with the normalization of decreased body weight and restoration of brain histopathological alterations. Conclusion: The results indicate that changes in depressive behavior and body weight are associated with remarkable modulation of oxidative stress and antioxidant defenses. Since oxidative stress triggers/exacerbates several routes of damage contributing to brain dysfunction and this could be an important therapeutic target for stress-related disorders like depression.

IL-8.

Antioxidant Potential and LDL Oxidation in Disease.

Dr. Neelima Singh and Nivedita Singh.

Department of Biochemistry, G.R. Medical College Gwalior (M.P) LDL particles are rich in antioxidants and are resistant to oxidation. The capacity which prevents the oxidation of LDL by free radicals is known as antioxidant potential (AOP). Diabetics are at higher risk of atherosclerotic cardiovascular diseases (ASCVD) than non diabetic. Present study which is carried out in 20 healthy control,15

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regulated D.M (by medicines) and 20 unregulated diabetic mellitus subjects reveals that sensitivity for LDL oxidation is increased significantly in unregulated diabetes mellitus subjects (P<.001) as compared to regulated diabetic group and control group along with reduced antioxidant potential (P<.001).

Indian Journal of Clinical Biochemistry Supplement, 2009/24

IL-9.

Total Quality Management in Health Laboratories

Dr. Ghassan Shannan

IFCC Treasurer, E-Mail: [email protected] Total Quality Management Concepts in Health Laboratories became a necessity in analytical laboratories. In USA and Europe various standards are used to implement and enforce Quality Management. However, the basic criteria for TQM are similar on both sides of the Atlantic. This paper will address the principles of TQM, International Standards, and Review of International Organisation Guidelines. Competence, Safety, Decision Making, Quality, Reliability, Precision, Accuracy, Contracting, Continuous Education, Traceability, Validation & Interpretation of Results all are part of any TQM programme. It is in the interest of Patients, Society, Health providers, Insurance Companies and Governments that Medical Laboratories performance is of High Standards of Professional and Technical Competence, due to the followings: Diagnosis, Prognosis and Treatment of various diseases are frequently based on the results and interpretations of Laboratory Tests. Patients and Clinicians may not have sufficient technical knowledge to determine whether a laboratory operates at satisfactory level. In many cases patients and/or clinicians have no choice of laboratory. IL-10. Laboratory Tests are expensive and they are expected to provide valid information. It is of the interest of competent laboratories to verify their competence through a process of inspection. TQM Process should include all laboratories and in-vitro diagnostic activities including physician office laboratories and point-of-care testing. TQM should include regular inspection of: 1. Premises & other Facilities 2. Qualification of Lab Directors. 3. Qualification of Technical Staff. 4. Equipment. 5. Organisation & Administration. 6. Ethics. 7. Quality Policy. TQM may be achieved by implementing ISO standards of which 15189 ­ 2007 is the more recent ISO dedicated for health laboratories. ISO 15189 addresses various aspects of Medical Laboratory Practice which include: 1. Scope of the standard. 2. Normative references. 3. Terms and definitions. 4. Management requirement.

Beyond Means and SDS: Recommended Statistics for Knowledge Extraction from Accumulated Pathology Data and Your Practice Experience

T F Hartley

Quality Manager, Pathology Services, Royal Hobart Hospital, Hobart, Tasmania, Austalia.. Email: [email protected] Objective : Clinical Biochemists have been largely focussed on parametric statistical tests such as means, standard deviations, coefficients of variation, linear regression and correlation coefficients. These tests are suited only to parametric data which conform to the criterion that the data analysed have been drawn from a notional `larger' parent dataset that when plotted as a

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histogram conforms to the `Normal Error Curve'. In this paper the focus is on those statistical tests which can be used on datasets upon which we have no way of confirming that they are part of a " `larger' parent `Normal Error Curve' ". This is particularly true of datasets from diseased patient populations. Such datasets are more probably skewed due to disproportionate mixing of data from patients with `mild', `moderate' and `severe'. Rather than risk applying invalid parametric statistical tests to such datasets and thereby being led into making misleading conclusions, the recommendation of this paper is that more use is made of non-parametric statistical tests. By going down this non-parametric path we can enter the new field of network analysis ­ the foundation of `knowledge mining' ­ and discover new and unexpected interdependences in our datasets. Methods: Wherever possible either Excel or web based calculators have been used to illustrate performance and the advantages. Non-mainstream Statistics Geometric Mean Regression, Rank Order Correlation, Logistic Regression; Statistics for Small Studies Chi Squares Mann Whitney U Test; Statistics for Knowledge Mining ANOVA, Repeated Measures ANOVA Multiple Linear Regression CHAID Analysis Bayes Theorem Bayesian Analysis of Questionnaire Data using Bayesware Discoverer Results: Results from studies using each of these statistical tests will be presented. Three of the most interesting results were in the relation to Geometric Mean Regression, Multiple Linear Regression and the Bayesian Network analyses. Geometric Mean Regression has been used to examine the relative performance of three local laboratories (y) and 93 National laboratories (x) in an External QA program for TIBC analyses. Y on x and x on y regressions both suggests either a + 8% or a -8% systematic bias depending upon your point of view, local or National. The geometric mean regression analysis however suggests that the consensus agreement should be

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that the local labs are reading 8% lower than the National labs. y on x regression : Local = 1.079 * National ­ 0.3537 x on y regression : National = 0.9236 * Local + 0.5783 Geometric Mean Regresion : Local = 1.081 * National - 0.4881 Multiple Linear Regression has been used to derive an equation that predicts the ionized Ca concentration in plasma : Calculated Ca2+ = 0.423 * Total Ca + 0.690, - 0.00620 * Albumin - 0.00149 Globulins, - 0.00257 Bicarbonate - 0.00433 * Anion Gap, - 0.033 * Phosphate (All parameters are in SI units). The Bayesian Network analysis of data from two cohorts of medical inpatients ­ one before the changeover from a Vitros 950 to an Abbott Architect and one after revealed the following `rules' based on the assumption that the clinicians did change over to the new reference intervals : They would form the impression that there were more low and fewer high Na and bicarbonate results. 77% of patients with high K's would also have high ureas. Only 55% of patients would have normal ureas and K's. 59% of patients would have low K's and normal ureas. 90% of patients with normal creatinines would have low ureas 33% of patients with normal creatinines would have high ureas. Conclusion: Now is an opportune time to expand our use of statistics in the clinical laboratory. We have particularly stable analytical platforms so we should spend more time on the objective examination of the very large amounts of data these systems produce. Within these datasets is a rich resource of knowledge that can only be appreciated after the application of robust and multiparameter statistical tests that go beyond our usual practice.

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IL-11.

Web Based Education and Training in Clinical Laboratory Services

Dr. Uday Donde

Director, UXL Advisor, Mumbai Web based education, a powerful tool in the education system, can help in imparting defined knowledge, conducting assessment regarding knowledge absorbed in terms of feedback and examination, and developing skills through animation. A simple platform to log on through a personal computer having internet connectivity, and registration of the user giving his specific requirements is imperative. Specific identification or password is given by the service provider to begin the process of imparting education. Several options are offered and a variety of software is available. A report card or performance rating can be generated to strengthen the student's efforts. As the student progresses, difficult level of the examination can be increased, in order to create further interest and increase knowledge. The advantage of web based education is that a student can learn what he wants, even from home, at his own pace, whenever he wishes. (The "whatever, whenever, wherever, however" concept). It is advisable to supplement web based education with on bench or in-field training in the field of clinical laboratory services. This can best be achieved by using animation / computer simulation which should be corroborated with actual practical work by providing self generated standard operating procedures. This computer aided comprehensive teaching ­ learning system can be employed to any profession teaching, clinical laboratory services, entrepreneurship etc. Besides web based education in clinical biochemistry web based awareness on regulatory bodies governing the discipline of clinical biochemistry can also be achieved. Salient information regarding the nature of governance / regulation can be communicated by providing links. The examples of such bodies are IS / ISO, Government Ministries per se, FDA, ICMR, MCI, NABL, Pollution Control Board, etc. A handful of IT companies have shown interest in developing this field for training clinical biochemistry professional using virtual classroom and e- learning methods. This would add value to the academic and applied components of this discipline. An exposure to this concept will be presente

IL-12.

Lean - Sigma Metrics Protocol of Total Quality Management in a New Laboratory Set-up

Barnali Das

Kokilaben Dhirubhai Ambani Hospital & Medical Research Institute, Mumbai. Objective: We aimed for Lean-Total Quality Management (TQM) for setting up the Biochemistry & Immunology laboratory for a 725 bedded new tertiary care hospital in Mumbai. Methods: Components of TQM include all divisions of the organization, namely, laboratory operations, information management, documentation and record maintenance, materials

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and purchase, customer care, safety etc. During the initial phase, we validated all the techniques and new tests by verifying reference intervals, analytical accuracy and precision, inter-assay and intra-assay variations, analytical sensitivity, linearity and reportable range. Results: We take the example of alkaline phosphatase (ALP) processed during start-up phase of our laboratory to enumerate the above validation steps. Reference range obtained for ALP was 42.2 - 108.8 IU/L, which is within the manufacturer's reference range (30 - 120 IU/L). Comparison of analytical accuracy between two laboratories gave, y = 0.98x + 4.93, with r2 = 0.99, establishing a high degree of linear correlation, validating our laboratory procedures. Precision verification from 15 replicates gave a coefficient of variation (C.V.) of 1.04%, comparable with the manufacturer's claim. Intra- assay variations were also within acceptable range (C.V. = 1.54%). 20 blanks were all within the stated blank value,

implying analytical sensitivity. Analytical measurement range (AMR) was verified as different levels of control sera were comparable with the manufacturer's claim, with C.V. 1.26% and 0.69%, for level I and level II, respectively. The standardized test showed good linearity with different dilutions; recovery too was within an acceptable limit (90-110%), with y = 0.99x ­ 2.81 and r2 = 0.99 between expected and observed values. Conclusion: We designed the entire loop of testing process, including pre-analytical, analytical & post-analytical phases according to the lean principle through pneumatic system and sigma metrics through DMAIC principle, i.e., Define, Measure, Analyze, Improve and Control. This and validation eliminates error in test results which is important from the view point of both patients, as well as of professional and regulatory bodies in the health care domain.

IL-13.

Laboratory Management of the Metabolic Syndrome

Sridevi Devaraj

Professor, Pathology, Director, Special Chemistry and Toxicology, University of California Davis Medical Center, Sacramento, California, USA; [email protected] The metabolic syndrome afflicts approximately 1 in 4 American adults and is prevalent in many industrialized countries. Furthermore, the prevalence of the metabolic syndrome is on the increase in Asian Indians. The metabolic syndrome confers at least a 2 fold risk of cardiovascular disease and at least 5 fold increased risk for subsequent diabetes. The features of the metabolic syndrome include abdominal obesity, an atherogenic dyslipidemia manifesting as elevated levels of triglycerides, low levels of HDL-C, and a preponderance of small dense LDL particles. Other features include raised blood pressure, insulin resistance and/or glucose tolerance, and associated increased

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inflammation. The laboratory plays a critical role in the diagnosis of the metabolic syndrome since of the 5 features, 3 are laboratory based, including triglycerides e"150mg/dL (1.7mmol/L), HDL-C levels <40 mg/dL (1.1mmol/L) and 50 mg/dL (1.3mmol/L) ( in men and women respectively) and fasting plasma glucose e"100mg/dL (5.6mmol/ L) . In addition, it is not uncommon for patients with metabolic syndrome to have non-alcoholic steatosis hepatitis (NASH), generally diagnosed by an increased in transaminases with ALT/AST ratio >1 and sonographic evidence of fat accumulation in the liver. Also, patients with the metabolic syndrome can have hyperuricemia and

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microalbuminuria. With regards to the atherogenic dyslipidemia since the laboratory reports in a standardized fashion triglycerides, HDL and LDL cholesterol, it is also incumbent on the clinical laboratory to report the non-HDL-C which embraces all the atherogenic ApoB carrying particles. It is simply obtained by subtracting HDL cholesterol from the total cholesterol. The non-HDL-C is especially important in patients with triglycerides e"200mg/dL (2.25mmol/L) and the goal for nonHDL-C is the LDL goal plus 30mg/dL (0.77mmol/ L). Thus, in a patients with diabetes and metabolic syndrome, a LDL-C goal would be <100mg/ dL (2.6mmol/L) and non-HDL-C would be <130mg/dL (3.3mmol/L). Other abnormalities that have been reported in the metabolic syndrome include elevated levels of remnant and remnant like particles and small dense LDL. Also, patients with the metabolic syndrome have elevated levels of ApoB, which is a better predictor of outcomes in statin trials than LDL-C. Microalbuminuria is also relatively easily measured in the clinical laboratory and indeed can be done on a spot urine sample, expressing it as albumin/creatinine ratio. Microalbuminuria defined as between 30 to 300 ug/mg of creatinine would

also encourage the clinician to better manage the patient especially with regards to hypertension and abnormal glucose tolerance. C-reactive protein (CRP) levels have been shown to predict cardiovascular events when measured by the high sensitivity (hs) assay and in fact are elevated in patients with the metabolic syndrome and appears to confirm greater CVD risk . A hsCRP level in patients with the metabolic syndrome between 3-10 mg/L could inform the clinician to better target the prothrombotic/proinflammatory status of the patient with therapeutic lifestyle changes and pharmacotherapy. Another very important adipokine that has great relevance to the metabolic syndrome is decreased levels of adiponectin. Lastly, whilst, some measure of insulin sensitivity can be obtained by using the homeostatic model (HOMA-IR), insulin assays are not standardized and thus any recommendation for reporting HOMA or QUICKI measures of insulin resistance requires better standardization for clinical diagnosis. Other circulating biomarkers such interleukin-6 (IL-6), monocyte chemotactic protein-1 (MCP-1), leptin, PAI1 retinol binding protein-4, may be relevant in metabolic syndrome but belong to the research arena presently.

IL-14.

Thyrotropin Secreting Pituitary Adenomas (TSHoma)

V.Parameswaran, G.Stilwell and J.Burgess

Diabetes and Endocrinology Services, Royal Hobart Hospital, Tasmania, Australia Thyrotropin (TSH) secreting tumours of the pituitary are uncommon and account for <1% of all pituitary tumours. Although, TSHoma are a rare cause of hyperthyroidism, it should still be ruled out in the diagnosis of hyperthyroid patients. TSH secreting tumours are slow growing and consequently establishing a positive diagnosis may take several years. Compared to males, females have a slightly higher rate (55%) of these tumours Laboratory results for thyroid function can be variable from normal to very high results. Radiological evaluation is required and exclusion of other biochemical abnormalities is critical in establishing a positive diagnosis of TSHoma. The steps involved in differential diagnosis will be presented and discussed using a clinical case.

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IL-15.

Monogenic Disease in Endocrinology

Tjin-Shing Jap

Section of Biochemistry, Taipei Veterans General Hospital, Taiwan, ROC 112 Monogenic disease is relatively quite common in the field of Diabetology and Endocrinology. Maturity-onset diabetes of the young (MODY) is responsible for only 2-5 % of type 2 diabetes in western population. It is also an autosomal dominant disorder with a typical age of onset of less than 25. Recently, we have identified two patients with MODY3 had missense mutations in exon 3 of the HNF-1á gene (Y218C and R272H, respectively) in a region of the protein that corresponds to a predicted DNA binding domain. Homozygous-trait hypercholesterolemia is due to mutation of gene encoding LDL-receptor, may cause severe xanthomatosis and early death by the age of 30. We identified 5 missense of mutations and one large deletion in LDLR gene, including one novel mutation in Taiwan. Mutation of gene encoding lipoprotein lipase may cause family hypertriglyceridemia with clinical manifestations of pancreatitis and early atherosclerosis. L252V mutation is the most common in Taiwan. Complete TBG deficiency may be misdiagnosed as a case of clinical hypothyroidism. We have found two cases with complete TBGD due to novel TBG gene mutations (S52N and W280Y). Familial hypocalciuria hypercalcemia is due to a mutation of the gene encoding calcium sensing receptor. We have reported a case with the long-standing benign hypercalcemia due to R648X mutation of CaSR gene. To our knowledge, it is the first case of FHH reported with CaSR gene mutation in China and Taiwan. We have identified the MEN1 gene mutation of nine Han Chinese families with MEN1 living in Taiwan, including five novel mutations. Recently we have also found a patient with CAH due to a compound heterozygous mutations of R362C and P409R in the P450c17 (17-??hydroxylase/ 17,20 Lyase) Gene. In recent years, we have successfully identified several families with clear single gene mutation causing hypercholesterolemia, Familial Hypocalciuric Hypercalcemia, Thyroid binding globulin deficiency, Alpha-lipoproteinemia, Familial hypertriglyceridemia and MODY, MEN1, Pendred syndrome and Congenital Adrenal Hyperplasia, respectively.

IL-16.

Obesity ­ A Global Epidemic

Dr. Nalini Ganesan

Professor, Department of Biochemistry, SRMC & RI, Sri Ramachandra University, Porur, Chennai ­ 116 The growing prevalence of obesity worldwide is an increasing concern surrounding the rising rates of type-2 diabetes, cardiovascular disease and consequent health and financial implications for the population. The United States has the highest prevalence rates of obesity worldwide, now exceeding 30 %. Moreover, the prevalence of obesity is rising in developing countries. The

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etiology of obesity is multifactorial and rapid rise in prevalence of this epidemic throughout the world has indicated that the environmental changes are the major determinants modifying the individual susceptibility to these environmental factors. The root cause for obesity being energy imbalance and is associated with significant morbidity and mortality. The classical problems associated with

Indian Journal of Clinical Biochemistry Supplement, 2009/24

obesity range from mechanical such as osteoarthritis and sleep apnea to metabolic perturbations resulting in type 2 diabetes, hyperlipidemia, hypertension and overall increased cardiovascular risk. There is also an increasing realization that obesity is associated with higher risk for developing certain cancers. This review addresses physiological and neuromolecular mechanism regulating energy balance by focusing on adipocyte metabolism,

adipocyte dysfunction, etiopathogenesis, environmental and genetic determinants, mitochondrial dysfunction and secondary pathologies promoted by obesity. Greater insight into mechanism behind this may improve our understanding of how to prevent and best manage this complex condition. We are planning to produce our experimental evidence on the biochemical and genetic aspect pertaining to obesity.

IL-17.

Novel Interactive Signal Molecules in the Hypothalamus Regulating GNRH Release

Dr. Edathil Vijayan

Centre for Neuroscience, Dept of Biotechnology, Cochin University of Science &Technology, Kochi -682 022 Kerala.. The human brain has mystified people throughout history. Though it weighs a mere 1.5kg and is small enough to hold in our hands, it is our body's most vital organ. It's complex network of 100 billion or more nerve cells orchestrates every aspect of our thoughts, perception and behavior. More than anything else, our brain defines who we are. Brain is remarkably plastic and continues to change throughout life in accordance with our experiences. Brain, behavior and environment are all intricately linked in an interactive loop: changes in the environment leads to changes in behavior which lead to changes in the brain. Immune cells which constitute the body's biological defense against infections and toxins have many things in common with nerve cells. One of the most exciting findings in recent years is that neuroendocrine system and immune system communicate through common biochemical language. Cytokines released from cells of immune system inhibit gonadotropin releasing hormone (GnRH) release an action similar to that of nitric oxide synthase (NOS) inhibitor. Nitric oxide (NO), an ubiquitous signal molecule, plays a crucial role in a host of biological systems including the brain as a neurotransmitter. Nitric oxide synthase (NOS) containing neurons occur in the hypothalamus where the perikarya are located in paraventriculr and supraoptic nuclei their axons project to the neural lobe of the pituitary gland , the structure that contain the largest quantity of NOS of any organ in the body. When both NOS inhibitor and cytokine were added together, both in vivo and in vitro incubation systems there was an additive suppressive effect on GnRH/ LH release. Our understanding of these complex interactions between the brain and the immune system is revealing targets for therapeutic intervention. For instance, possibility of various vaccines that might arrest or slow the progress of brain tumor growth or Alzheimer's disease and clinical trials of an immune therapy for spinal cord injury are underway. (Supported by ICMR, India).

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IL-18.

Role of Ethanol in Inducing Inflammatory Brain Damage and Vasculogenesis

Sukhes Mukherjee #, Subir Kumar Das$, Subrata Chattopadhyay, and D.M Vasudevan #

Department of Biochemistry, Amrita School of Medicine, Kochi-682026, $ Department of Biochemistry, Agartala Govt Medical College, Kunjaban P.O., Agartala 799 006, * Bio-organic Division, Bhabha Atomic Research Centre, Mumbai 400 085; [email protected] Background: Ethanol is a small molecule that has a negative impact on human health and moderate to heavy doses of alcohol have deleterious effects and especially on brain and its different compartments. Ethanol is also responsible for degenerative changes in brain. Methods: In the present study with Male Wistar rat, we investigated the role of natural antioxidants like grape polyphenols and Vit E in combating the ROS balance in alcoholism for a dose of 4g / Kg b.w/day of ethanol for a period of 12 weeks. Results: Ethanol intoxication in brain upregulates COX-2, iNOS levels and p38 MAPK pathways as evidenced by immunohistochemical studies of different sections of brain, and increases cell death in different brain compartments as evidenced by Caspase-3 and Caspase-8 expression and TUNEL assay by Immunohistochemistry. Conclusion: Grape polyphenols show protective effect to ethanol induced brain damage by regulating the inflammatory processes with the alteration of the VEGF receptors (flk1 and flt1) and VWF-VIII facilitating vasculogenesis. These findings indicate that exposure to ethanol for a higher dose induces brain damage triggering the inflammatory processes and leads to neuronal apoptosis.

IL-19.

Culture of Rat Gastric Mucosal Cell Line RGM1 or its Tumor Cell Line RGK1 Combined with Human Fibroblasts

Hideyuki J Majima1), Yoko Yamaguchi2), Mitsuhiro Ohshima2), Hiroko P Indo1), Kazuo Tomita1), Yoichiro Iwashita1), Hsiu-Chuan Yen3), Takuro Kodama1), Masakazu Souda1), Akihide Tanimoto1), Motoharu Miymoto1,4), Kazuyuki Noguchi4), Osamu Shimokawa5) Yumiko Nagano5), Tsuyoshi Kaneko5), Hirofumi Matsui5), and Toshihiko Ozawa6),

Department of Oncology, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima 890-8544, Japan, 2Department of Biochemistry, Nihon University School of Dentistry, Kanda 101-8310, Japan, 3)Graduate Institute of Medical Biotechnology, Chang Gung University, Kwei-Shan, TaoYuan 333, Taiwan, 4)Department of Periodontology, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima 890-8544, Japan, 5)Division of Gastroenterology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan, 6)Department of Health Pharmacy, Yokohama College of Pharmacy, Totsuka 245-0066 Japan. Hideyuki J Majima: [email protected] Studies using cell culture have benefits to provide bioenvironmental condition or factors. However, selective data excluding complex sometimes the data will not reflect in vivo

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1)

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conditions due to lack of physiological or pathological conditions where interactions of different types of cells would exist. Besides in vivo, in vitro 3D cultures with different types, e.g., epithelial cells and fibrobasts will provides new aspects of interactions between the two types of cells. A rat normal gastric mucosal cell line RGM1 or its tomorized cell line RGK1 were used to perform 3D culture with human fibroblats. Human fibroblasts were cultured in collagen type I and RGM1 or RGK1 were cultured on the collagen as 3D cultures. 3D cultures with RGM1 or RGK1 cells with only collagen resulted monolayer

growth on collagen, whereas those cell 3D cultures with human fibroblasts showed appearances of activation in both the epithelial cells and fibroblasts, suggesting the interactivity between the two types of cells. Particularly, in RGK1 cells grew with multiple cell layers (the layers), and fibers and fibrous margin were found inside the layers and in collagen site facing the layers. Results by treatment of a matrix metalloproteinase (MMP) inhibitor using Marimastat to the 3D cultures showed disappearance of activation of both the epithelial cells and fibroblasts or dissociation of the interactivity.

IL-20.

Cancer Biomarkers of Prognostic Value

Paturu Kondaiah

Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore 560 012. E-mail: [email protected] The field of Biomarkers that can predict the course of the cancer progression has been an important area of intense research in recent times. Gene expression profiling using gene microarrays has been the choice of investigation for a high throughput analysis and identification of biomarkers. Our group has been working with cancer and pre cancer tissues to identify novel biomarkers of prognostic value. We carried out microarray analysis of glioma tumors of different grades using cDNA microarray. Data analysis identified several differentially regulated genes between normal brain tissue and various grades of gliomas. Further, several novel genes are differentially expressed between grades II/III astrocytoma (low grade) and glioblastoma multiforme (GBM; grade IV). By real time PCR analysis we found differential regulation of several genes in the Notch signalling pathway such as ASCL1 (Hash 1), Hes1, Hes6 and Delta1. In addition, differential expression of many other already known and novel genes in gliomas has been identified. Real time PCR validations of some selected genes are in agreement with the array results. Genes that show differential regulation are in the categories of transcription factors, signalling intermediates and secreted factors. In addition, we found expression of few genes correlating with prognosis of the GBM patients. Our data could form a basis for building diagnostic/ prognostic gene signatures in glioma progression and novel therapeutic targets.

IL-21.

HPV 16 Variants in Normal, SIL and Cervical Cancer Women.

Neeta Singh1, Chandresh Sharma1, Archna Singh1 Palika Datta1 and Sunesh K. Jain2

Departments of Biochemistry1, Obstetrics and Gynecology2, AIIMS, New Delhi ­ 110029, India. E-mail: [email protected] Background: HPV is a causative factor in cervical carcinogenesis with HPV 16 being the most prevalent. E6 and E7 proteins are associated with transformation and immortalization, whereas L1

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is a capsid protein responsible for virus assembly. Objective: We investigated variants prevalent in HPV 16 E6, E7, L1 genes in Northern India, in normal, SILs and cervical carcinomas and their possible biological effects.

region. Some of these led to amino acid changes such as R10G, Q14H, D25N, D25E, S71C, H784Y, and the most prevalent being L83V. E7 showed a total of 7 variants, with F57V being the most prevalent. L1 showed a maximum of 58 variants Methods: Cervical scrapes/biopsies were in tumors, with Ins 448S and D465 Del being the collected, DNA extracted and examined for the most prevalent. presence of HPV by PCR using consensus MY09/ Conclusion: Similar variants were seen in 11 primers and for HPV 16 using type specific normals, SILs and cervical carcinomas, but to primers. HPV 16 variants were characterized by varying extents. The identified variants some new full length sequencing of L1, E6 and E7 genes. and some already reported can disrupt virus Results: 89% of normals, 100% of SILs and 71% of HPV 16 positive tumors showed the presence of the European variant class, mainly E350G and E350T. A total of 18 variants were seen in E6 assembly and transcription, P53 degradation, B and T cell epitopes and are thus important in viral persistence and progression to cervical cancer, and also in vaccine development

IL-22.

An Update in Molecular Oncology: Cervical Cancer

B.Nagarajan

Cancer Institute Chennai- 600 020, Email: [email protected] In the Asia-pacific region incidence of cervical cancer cervix is high second to breast cancer. Prognosis of invasive carcinoma is based mainly on subjective clinical staging. Abnormalities in cytokine production have been found in invasive cervical carcinomas. A correlation between cytokine expression and different clinical parameters such as tumor and lymph node status, HPV expression and disease free survival were evaluated. Human Papilloma Virus (HPV) is the major cause. A total of 43 cervical biopsies collected from sequentially enrolled patients were analyzed by DNA amplification and sequenced to determine the HPV genotype. Samples with multiple infections were resolved by multiplex PCR combined with array primer extension (APEX) Nine different HPVs mainly 16 and 18 were detected. Multiple HPV infections correlated prominently with lack of treatment response compared with single type infection that again reiterates the risk of treatment failure. Cytokine

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studies address macrophages and dendrite cell infiltration, role of IL-17, IL-6 and 8 as prognostic markers, levels of both T helper cells 1 and2 cytokines and T cell anergy. Furthermore DNA adducts represent an early detectable and critical step in carcinogenesis and may serve as an internal dosimetry of carcinogen exposure. To determine changes in oxidative and other DNA adducts. During cervical cancer development, we analyzed DNA from specimens collected from cervicites, biopsy-proven dysplasia and invasive cancer cases. These toxic biological end-points could as well form therapeutic indices in the management. An offshoot of this approach is to monitor DNA adducts levels in patients undergoing radiotherapy. Our data tend to support the commonalityinfection, inflammation and release of ROS. The bottom line is prognosticate and stratify patient response based on data base that would end result in ultimate patient care and viable point-of-care diagnostics.

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IL-23.

Natural Products as Potential Treatments for the Human Metabolic Syndrome

Lindsay Brown

School of Biomedical Sciences, The University of Queensland, Australia Metabolic syndrome is the clustering of abdominal obesity, insulin resistance, dyslipidaemia, hyperglycaemia, a prothrombotic state and hypertension that increases the risk of cardiovascular disease and fatty liver in humans. Diets rich in carbohydrates and saturated fats contribute to the signs of the metabolic syndrome. Animal models that mimic the range of pathophysiological changes seen in humans diagnosed with metabolic syndrome are necessary to understand the initiation and progression of metabolic syndrome and the development of new treatments. The metabolic changes and the structure and function of the heart and liver were characterised in young adult male Wistar rats fed a high carbohydrate/high fat diet for 16 weeks. These rats developed many of the chronic signs of the metabolic syndrome in humans including hypertension, impaired glucose tolerance, increased oxidative stress, plasma lipid abnormalities, excess deposition of abdominal visceral fat, increased waist circumference, ventricular dilatation with hypertrophy and decreased contractile function, impaired conduction, increased inflammatory cell infiltration into the myocardium and liver resulting in fibrosis, an increased stiffness of the heart, endothelial dysfunction as a decreased response to acetylcholine and increased plasma concentrations of liver enzymes. Further, oral chronic treatment with curcumin or rutin attenuated or reversed these structural and functional changes in the heart and liver. Thus, this dietaryinduced model of the metabolic syndrome in rats may provide a realistic and useful experimental tool to evaluate the development of symptoms and their reversal by potential treatments such as natural products.

IL-24.

Association of Cytokines and Insulin Resistance in Non-Alcoholic Fatty Liver Disease

Subir Kumar Das

Department of Biochemistry, Agartala Govt Medical College, Kunjaban P.O., Agartala 799 006, Tripura, e-mail: [email protected] Background: A number of factors are linked with non-alcoholic fatty liver diseases (NAFLD), a condition that ranges from clinically benign fatty liver to its more severe form, non alcoholic steatohepatitis (NASH). Objective: To evaluate the role of anthropometric profile, lipid profile, insulin resistance data, proinflammatory and anti-inflammatory cytokines in NAFLD patients. Methods: A complete clinical history, including body mass index and waist-to-hip ratio were obtained in 105 NAFLD patients (89 male and 16 female) and 77 normal subjects (45 male and 32 female). Blood collected after over night fast was

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used for the determination of biochemical parameters including glucose level, insulin level, lipid profile and cytokine estimations. Insulin resistance was calculated by the homeostasis model. Results: Predisposing factors were diabetes mellitus (35%), overweight (56%) and hyperlipidemia (44%). Insulin resistance was confirmed in 70% NAFLD patients, and 42% fulfilled minimum criteria for insulin resistance syndrome. NAFLD patients showed elevated

levels of TNF-a and TGF-b1, while IL-4 level decreased significantly compared to normal subjects. Conclusion: Insulin resistance and cytokines may be responsible for fibrogenic processes in NAFLD. Acknowledgement: Financial assistance received from Van Slyke Foundation- Critical and Pointof-Care Testing (VSF-CPOCT) Research Grant; American Association for Clinical Chemistry is gratefully acknowledged.

IL-25.

The Promise of Stem Cells and Gene Therapy as Novel Approaches to the Treatment of Multiple Sclerosis (MS)

Natalie Payne1, Christopher Siatskas1, Guizhi Sun1, Daniella Herszfeld1, Natalie Seach1, Sara Litwak1, Martin Short1, Ban-Hock Toh2,Frank Alderuccio3, Thomas Backstrom4, Richard L Boyd1and Claude C A Bernard1

Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Victoria, 3800, Australia. 2Centre for Inflammatory Diseases, Department of Medicine, Monash University, Clayton, Victoria, 3168, Australia. 3Department of Immunology, Monash University, Melbourne, Victoria 3004, Australia. 4Malaghan Institute of Medical Research, Wellington, 6012, New Zealand.

1

Multiple sclerosis (MS) is a disease that affects the central nervous system (CNS) with significant human and community costs. Although the precise trigger(s) remain elusive, the ultimate end point of the disease is the destruction of nerve axons and myelin, two key cellular components of the CNS involved in the conduction of nerve impulses. As a consequence patients with MS suffer from numerous neurological problems. At present, there is no cure for this severe

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neurodegenerative condition. Although, immune therapy is beneficial to a proportion of patients, others do not respond to any currently available therapies. A rational approach to the treatment and cure of autoimmune diseases such as MS requires incorporation of three fundamental processes: suppression of the inflammatory response, regeneration of the target cells or tissues and restoration of self tolerance, whereby damaging white immune cells can be purged or silence. Current clinical

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practice at best addresses the first problem but there are no clear strategies for the later two. To address these issues we are using experimental autoimmune encephalomyelitis (EAE), a model for MS, induced by immunization with the myelin oligodendrocyte glycoprotein (MOG), a key autoantigen in MS. Due to their multipotentiality, broad immunomodulatory properties and potential to facilitate repair of damaged tissue, mesenchymal stem cells (MSC) hold great promise for treatment of central nervous system and other diseases. While MSC were originally isolated from bone marrow, it is now recognised that many connective tissues contain a population of MSC. In this study, the therapeutic potential of human MSC isolated from bone marrow, adipose tissue and umbilical cord Wharton's jelly was investigated using the EAE model. All three cell types were initially characterised by their differentiation ability, colony forming unit-fibroblast assays and by flow cytometry using an extensive panel of antibodies. Immunomodulatory properties were also assessed in vitro through T cell proliferation assays. We next examined the effect of MSC on the development of chronic progressive EAE induced by the extracellular domain of MOG (rMOG). While BM-MSC appeared to have little effect on the disease course, disease onset was significantly delayed in mice receiving either A-MSC or UMSC. Maximum clinical score and cumulative score were reduced in A-MSC treated mice compared to the vehicle control group. Likewise, histological examination revealed that inflammation, demyelination and axonal loss were also reduced

in these mice. These findings demonstrate that MSC isolated from different tissue sources have distinct effects both in vitro as well as in vivo. Having demonstrated that MSCs have the ability to modulate the pathogenic immune response in EAE and thus might be of potential therapeutic relevance to MS, we next attempted to restore immune tolerance to MOG by intrathymic injections of a lentiviral vector engineered to express MOG (LV-MOG) or the hydrogen potassium ATPase -an irrelevant autoantigen (LV-HKATPase) or PBS. Neurological assessment indicated that untreated and control animals injected with LVHKATPase or with PBS, developed fulminant symptoms of EAE. In contrast, animals receiving the therapeutic LV-MOG vector manifest little or no signs of disease. Histological analysis of CNS from control animals showed inflammatory lesions that correlated with severe areas of demyelination and axonal loss. Notably LV-MOG-treated animals demonstrated significantly fewer cellular infiltrates that corresponded with a lower number of demyelinating areas with little, or no axonal loss as compared to controls. In LV-MOG-treated animals, a reduction in the number of antigen-specific CD4+ T-cells was demonstrated. Furthermore, a significant reduction in Th1 and Th17 cytokine levels was also shown in these cultures. Collectively these data indicate that intrathymic delivery of a LV-MOG represents a novel form of tolerance induction for the treatment of MS. Collectively, these findings further add to the armamentarium of nontoxic gene-based strategies for the treatment of this debilitating disease.

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IL-26.

Hs-CRP, Inflammation and Its Role in Coronary Heart Disease

Arun Raizada1, H V Singh2 and Naresh Trehan3 Senior Consultant and Head, Ex-President ACBI, Department of Clinical Biochemistry, Medicity, Global Health Pvt. Ltd, Gurgaon-122001, 2 Biochemist, Department of Pathology, Hindu Rao Hospital, Delhi-110007, 3 Chairman and Managing Director, Medicity, Global Health Pvt. Ltd, Gurgaon. "Inflammation" is the process by which the body responds to injury or an infection. Laboratory evidence and findings from clinical and population studies suggest that inflammation is important in atherosclerosis. This is the process in which fatty deposits build up in the inner lining of arteries. C-reactive protein (CRP) is one of the acute phase proteins that increase during systemic inflammation. It's been suggested that testing CRP levels in the blood may be an additional way to assess cardiovascular disease risk. A more sensitive CRP test, called a highly sensitive C-reactive protein (hs-CRP) assay, is available to determine heart disease risk. Number of studies have examined whether hs-CRP can predict recurrent cardiovascular disease, stroke and death in different settings. High levels of hs-CRP consistently predict recurrent coronary events in patients with unstable angina and acute myocardial infarction IL-27. (heart attack). Higher hs-CRP levels also are associated with lower survival rates in these patients. Many studies have suggested that after adjusting for other prognostic factors, hs-CRP is useful as a risk predictor. Studies also suggest that higher levels of hs-CRP may increase the risk that an artery will reclose after it's been opened by balloon angioplasty. High levels of hs-CRP in the blood also seem to predict prognosis and recurrent events in patients with stroke or peripheral arterial disease. Scientific studies have found that the higher the hs-CRP levels, the higher the risk of having a heart attack. In fact, the risk for heart attack in people in the upper third of hs-CRP levels has been determined to be twice that of those whose hs-CRP level is in the lower third. The true independent association between hs-CRP and new cardiovascular events hasn't yet been established.

1

Pro-oxidant Activity of Artemisinin Causes an Apoptosis Like Death in Leishmania parasites

Mitali Chatterjee and Rupashree Sen Dept. of Pharmacology, Institute of Postgraduate Medical Education & Research, 244B, Acharya JC Bose Road, Kolkata Email: [email protected] Leishmaniasis is a vector-borne disease caused by the obligatory protozoan parasite, Leishmania sp., affecting over 12 million people worldwide. In India, the emerging refractoriness to conventional

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antimonial treatment has highlighted the need for newer antileishmanial compounds. Artemisinin, an established anti-malarial drug, was found to be an effective anti-leishmanial drug, the IC 50 in

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amastigotes being 22 µM. This leishmanicidal activity was mediated by an increased generation of reactive oxygen species, coupled with depletion of non protein thiols; furthermore, the addition of N-acetyl cytseine increased the IC 50 of Artemisinin, confirming the critical role of free radicals. This oxidative insult upon the parasite triggered an enhanced influx of Ca++, translating into mitochondrial membrane depolarization, increased annexin-V positivity, deoxynucleotidyltransferase-mediated dUTP nick end labeling, finally leading to an increased sub IL-28.

G0/G1 population, features of apoptosis. However, this programmed cell death was not accompanied by enhancement of caspase activity suggesting non-involvement of caspases. Additionally, in vivo experiments in L. donovani infected mice confirmed the efficacy of Artemisinin (10 and 25 mg/kg b.w.) which was accompanied by up regulation of intracellular levels of Th1 cytokines, IFN-ã and IL-2 in splenocytes. Taken together, Artemisinin is an effective leishmanicidal agent that causes an apoptosis like death in parasites, meriting future pharmacological studies.

Novel Filarial Genes or Recombinant Filarial Proteins as Effective Vaccine Candidates

MVR Reddy Department of biochemistry & JB Tropical Disease Research Centre, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Maharashtra -442102 (e-mail: [email protected]) The rapid progress in Filarial Genome Project has led to the identification of variety of novel filarial genes or recombinant proteins of Brugia malayi or Wuchereria bancrofti as promising vaccine candidates. These include abundantly expressed molecules viz., abundant larval transcripts (ALT) and venom allergen homologue (VAH), antioxidant enzymes viz., glutathione-Stransferase (GST), superoxide dismutase (SOD), thioredoxin peroxidise (TPX) and other crucial enzymes like transglutaminase (TGA). Vaccination with B. malayi enzymes SOD, TGA, TPX induced low to moderate (30 to 46% ) cytotoxicity against microfilaria (mf) or L3 larvae or protection against challenge B. malayi infection in jird model. In contrast Bm ALT-2 or Wb GST induced 69% and 61% protection respectively against B. malayi challenge infection in jird model. When compared to these single antigen vaccines, fusion proteins or multiple antigen vaccinations generally offered better protection. Bm ALT-2 in combination with Wb GST, Bm TPX and Bm VAH enhanced the protection to 71%, 74% and 78% respectively. Although DNA vaccines have several advantages over conventional vaccines, filarial DNA vaccines particularly in single plasmid vaccination were often not adequate in antibody production and protection. DNA vaccine of Bm-tga could not induce much protection against challenge infection in mouse model. DNA vaccines of Bm-vah, Bm alt-2 and Bm-tpx induced only 50.2%, 55.8% and 37% cytotoxicity respectively against B.malayi mf. In contrast the cocktail vaccination of Bm-alt-2 in combination with Bm-vah and Bm-tpx induced up to 63% and 78% cytotoxicity respectively against mf or L3 larvae. Taken together these studies favour the use of multiple antigen vaccination with combination of two or more antigens / genes as an effective vaccine development strategy in Filariasis.

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IL-29.

Bilirubin from Toxic Metabolite to an Antioxidant: Mechanisms

S. Adhikari Radiation and Photochemistry Division, Chemistry Group, Bhabha Atomic Research Centre Mumbai 400 085, India Email: [email protected] Bilirubun has two extreme roles to play in physiology. In one side it is a toxic metabolite while at micromolar concentration it is better antioxidant than???tocopherol and thiols. It is a toxin because it's association with neonatal jaundice, possibly leading to irreversible brain damage due to neurotoxicity. On the other hand, bilirubin can act as an important cytoprotector of tissues that are poorly equipped with antioxidant defence systems, including myocardium and nervous tissue. Bilirubin can suppress oxidation of lysosomes at oxygen concentrations that are physiologically relevant. The antioxidant-prooxidant mechanism of bilirubin will be discussed in the present paper. Our study demonstrates that hydroxyl and glutathiyl radicals abstract hydrogen atom from bilirubin and forms a carbon-centered radical. The bilirubin radical thus produced, decays with first order rate and decays faster in presence of oxygen. At a later stage bilirubin forms a peroxyl radical in presence of oxygen. This is the first direct observation of the bilirubin peroxyl radical formation which may answer the situation of low IL-30. protection of lipid peroxidation in a condition of higher than the physiologically relevant oxygen concentration. Bilirubin induced toxicity may also arise from the aforesaid peroxyl radical formation. Linoleic acid peroxyl radical also reacts with bilirubin via hydrogen abstraction. In the case of NO2. radical reaction, bilirubin produces radical cation via single electron transfer reaction. These results show that in the oxidation of bilirubin both hydrogen atom transfer and single electron transfer are possible depending on the nature of the oxidizing radical. HPLC analysis of the products also supports our mechanism. Another aspect of bilirubin that will be discussed is the free radical induced recyclization. Through its antioxidant action, bilirubin produces biliverdin as a major product. The conversion of biliverdin to bilirubin is a physiological enzymatic process. Results from our laboratory shows that under certain condition, this recycling process from bilirubin to biliverdin and again bilirubin back can also occur through a free radical pathway.

New Perspectives on the Clinical Importance and Applications of Vitamin D Testing

H.A Morris Hanson Institute, SA Pathology, Adelaide South Australia 5000 The vitamin D system plays a primary role in the maintenance of calcium, and phosphate homeostasis. Recently it has been identified as exerting a wider range of biological activities including regulation of cellular differentiation and proliferation and therefore it has implications for cancer, the immune system, and reproduction. The endocrine action of vitamin D acts through the

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renal metabolism of vitamin D, producing 1,25 dihydroxyvitamin D (1,25D) for the circulation. The major organ responding to plasma 1,25D is the small intestine where it controls intestinal absorption of calcium and phosphate. 1,25D is also synthesised in a wide range of tissues including bone cells where it is believed to exert an autocrine or paracrine activity. Vitamin D insufficiency in

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the elderly has clearly demonstrated an increased risk of hip fracture. A major question for the clinical laboratory is what is the vitamin D requirement for a healthy skeleton? Measurement of vitamin D status in apparently healthy individuals merely assesses their level of sunlight exposure and does not indicate vitamin D requirements. Current evidence from a wide range of sources including clinical data indicate that levels of serum 25-hydroxyvitamin D (25D) below 20 nmol/L are clearly deficient and give rise to IL-31.

osteomalacia, a bone mineralisation defect. However evidence is accumulating to suggest that 25D levels between 20 and 80 nmol/L indicate a depleted vitamin D status causing bone loss and increasing the risk of fracture as well as increasing the risk of colon cancer and infection. These findings suggest that the lower level of the desirable limit for vitamin D should be significantly higher than currently used by many laboratories

An Insight to the Mechanism of Lycopene: A Biocarotenoids Against ã-Radiation Induced Apoptosis; MAPK Signaling Pathways and DNA Repair Enzymes in Normal Lymphocytes and Lymphoblastoid TK-6 Cell Lines in Vitro.

Venugopal P. Menon Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar-608 002, Tamil Nadu, India. We evaluated the effect of lycopene a dietary carotenoid on human lymphocytes exposed to radiation. Cells were exposed to ã-radiation at a dose of 4 Gy in the presence and absence of lycopene pretreatment. Lymphocytes when exposed to ã-radiation at a dose of 4 Gy resulted in DNA damage, delay in cell cycle, DNA fragmentation, up-regulation of caspases-3 caspases 9, Bax, cytochrome-c and MAPK signaling pathways and down regulation of Bcl2, DNA repair enzymes (HOGG1 and XRCC1) and reduction in the mitochondrial transmembrane potential when compared to normal control. When these cells were pretreated with lycopene and then administered radiation all these effects were reversed. Suggesting that IL-32. lycopene offers protection to normal cells when exposed to radiation. We also wanted to study whether lycopene can offer protection to cancer cells when exposed to radiation. In this context we used lymphoblastoid TK-6 cells in culture. When TK-6 cells was pretreated with lycopene and then administered ã-radiation, we found that the intensity of damage to cancer cells were much higher when the same amount of radiation was give to control TK-6 cells. Thus the study shows that lycopene while protecting normal cells acts in combination with ã-radiation to destroy more amount of cancer cells than in control TK-6 cells given radiation.

The Role of CRP in Atherothrombosis

Ishwarlal Jialal, Professor of Internal Medicine(Endocrinology, Clinical Nutrition and Vascular Medicine), University of California Davis Medical Center , 4635 2nd Avenue , Research 1, Room 3000, Sacramento , CA 95817 , USA. Background: Inflammation is pivotal in all phases of atherosclerosis. Among the numerous

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inflammatory biomarkers, the largest amount of published data supports a role for high-sensitivity C-reactive protein (CRP) as a robust and independent risk marker in the prediction of adverse cardiovascular events in both primary and secondary prevention In addition to being a risk marker, there is much evidence indicating that CRP participates in atherogenesis. Results: In this presentation, I will focus on the role of CRP in promoting atherothrombosis by discussing its effects on endothelial cells, endothelial progenitor cells, monocytemacrophages, and smooth muscle cells. The most abundant data relates to its effects on endothelial cells; CRP inhibits eNOS via uncoupling, stimulates monocyte adhesion, PAI-1, IL-8, and IL-33.

M-CSF via activation of NF-KappaB. Most recently we have shown that it perturbs the glycalyx, which decorates the luminal surface, resulting in endothelial dysfunction. Many of the effects such as impairment of vasoreacitvity, stimulation of Tissue Factor, MMP-9 and increase uptake of oxidized-LDL have been confirmed invivo. Conclusions: CRP is clearly a risk marker for cardiovascular disease and is recommended for use in primary prevention. In addition, CRP appears also to contribute to atherogenesis. However, further in-vivo research is needed to establish its role in atherothrombosis. This will lead to novel therapeutic strategies.

The Clinically Effective Use of Available Cardiac Biomarkers.

Ray U Department of Pathology, Royal Hobart Hospital, University of Tasmania, Australia Heart is a non-stop pump supplying nutrition and oxygen to the cells of the body including its own through coronary circulations. Coronary circulation has its unique networks. Any derangement in its own supply would cause cellular anoxia, trigger cellular leakage and jeopardise its pumping function. Acute coronary syndrome (ACS) is diagnosed by elevated cardiac markers, ECG changes and acute chest pain/ heaviness in the chest. Although currently there is a strong suggestion to use only the 99 th percentile changes in serial cardiac Troponin in the diagnosis of ACS, judicious use of multiple cardiac markers (CK-MB, Myoglobin and cTroponin) could be useful in the early diagnosis

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of ACS so that timely institution of either thrombolytic or interventional therapy could be done to save this hard working indispensable pump. Any increase in the intraventricular pressure or volume or both stimulate cardiac myocytes which release B-type natriuretic peptide (BNP) detectable immediately in the circulation. It is quite probable that acute ischemic event alters heart pump and brings forth a change in intraventricular pressure and volume. Thus there should be BNP release in the circulation. Serial assay of BNP along with CK-MB, myoglobin and c-Troponin could be thus used not only to diagnose ACS but also to stratify risk associated with it and further prognosis following therapy.

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IL-34.

Mycobacterial ES-31 Serine Protease of Interest in Diagnosis of Tuberculosis and HIV-TB Co-infection and with Potential as Drug Target

B.C. Harinath JB Tropical Disease Research Centre, Mahatma Gandhi Institute of Medical Sciences, Sevagram, India, E-mail: [email protected] Tuberculosis (TB) remains a major threat to public health worldwide and has been declared as global emergency by WHO in 1993. According to the World Health Organization (WHO, 2009), 9 million new TB cases including 4 million smear positive cases with 15% HIV- Positivity have been reported in 2007. Seroreactivity of the purified excretory secretory (ES) antigens ES-31, ES-43, ES-41 and ES-6 have been assessed in pulmonary tuberculosis (fresh, relapse, chronic and latent), extrapulmonary tuberculosis and in HIV-TB co-infection. Analysis of immune response to these purified antigens by indirect and sandwich ELISA using highly sensitive and semiquantitative Penicillinase enzyme immuno assay, showed ES31 antigen as having good diagnostic potential in pulmonary tuberculosis and in certain groups of extrapulmonary tuberculosis in particular tuberculous lymphadenopathy, tuberculous meningitis, whereas ES-41 was found to be more seroreactive in abdominal and bone & joint tuberculosis. ES43 antigen was primarily recognized by serum antibodies in relapse cases while ES-6 was useful in contacts. Antigen assay was found to be more sensitive than antibody based assay for detecting tuberculosis with HIV coinfection. Immunomonitoring for presence of antigen in TB patients under ATT, showed that ES-31 antigen assay was useful in determining the effectiveness of therapy and patient's compliance. User friendly Peroxidase ELISA has been standardized for detection of circulating mycobacterial ES-31 serine protease (Free antigen and Immune complexed antigen) with 70-75% sensitivity and 90% specificity and with a limit of detection of antigen at 1 ng/2?l (0.5 ?g / ml serum). Further study revealed that ES-31 antigen is a zinc containing serine protease. Serine protease inhibitors such as pefabloc, 3, 4 dichloroisocoumarin, phenyl methyl sulphonyl fluoride (PMSF) and metalloprotease inhibitors such as ethylene diamine tetracetic acid (EDTA) and 1, 10 phenanthroline inhibited 65 - 92 % of serine protease activity in vitro. Isoniazid showed 95% inhibition on mycobacterial ES-31 serine protease. These inhibitors also showed decreased bacterial growth in axenic culture and inhibition is further confirmed by decreased amount of ES-31 serine protease in culture filtrate. In human macrophage culture, highly inhibitory pefabloc, 1,10 phenanthroline and isoniazid inhibited infectivity of virulent as well as avirulent M. tuberculosis bacilli to macrophages. The enhancement of bacterial growth in the presence of mycobacterial ES31 serine protease while inhibition in the presence of anti ES-31 serine protease antibody was observed showing the importance of enzyme for entry and multiplication of bacilli. Immunofluorescence study using Fluorescence labelled anti ES31 antibody showed the presence of ES-31 serine protease on mycobacterial cell surface.

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IL-35.

Autoantibodies: Is it Possible to Predict Autoimmune Diseases?

Marcos López-Hoyos Servicio Inmunología Hospital Universitario Marqués De Valdecilla, Santander, Spain. A good biomarker should fulfill four characteristics: sensitive, specific, detectable in early disease and predictive of prognosis. Autoantibodies are the most widely used biomarkers in the diagnosis of autoimmune diseases and, in some cases, in the prognosis prediction. In the last years several prospective studies have been published supporting their utility in the prediction of an ongoing autoimmune disease in healthy subjects, although they are still few and large scale studies are needed. The prediction of autoimmune disease is of special utility since autoimmune diseases can be preceded by a symptom-free phase that may last for many IL-36. years. In this scenario autoantibodies may have a high positive predictive value for the disease onset, severity and organ-specific complications, especially in genetically prone individuals, in whom an environmental trigger may switch on the disease. One of the most active areas of work at present in autoimmunity is the development of autoantibodies profiles to identify those patients at risk of developing an autoimmune disease. These autoantibody profiles can be applied to the clinical management of healthy population and high-risk subjects. The session will review the recent evidence on the predictive value of autoantibodies and will underline the future lines in this field.

Cell-Cycle Synchronization in Antinuclear Antibodies Detection

Petraki Munujos Vinyoles R&D Department. BioSystems, S.A. Barcelona, Spain Anti-nuclear antibodies (ANA) are one of the most common parameters analyzed in the autoimmunity laboratory, and the indirect immunofluorescence technique, the major screening method in systemic autoimmune diseases. The identification of the immunofluorescent staining patterns on HEp2 cells is often difficult due to the complexity of the interpretation of the images observed. In addition, the quality of the cell culture, the developing procedure and the conjugate, among others, are the key point for a correct determination. These cell cultures must meet some requirements to guarantee good results, among which the presence of cells in mitosis has become a differenctial feature when judging the suitability of the HEp2 cell slides manufacturers. There are mainly two reasons for which it is important a certain per76

centage of cells in mitosis: the existence of autoantibodies reacting with proteins that only express during the mitosis phases, and the fact that some patterns are similar to such an extent that the only distinctive feature to identify them is the staining shown by the cells in mitosis. While in an asynchronic culture, only up to 1-2% of cells are found in M phase, a culture that has been synchronized in early S phase can display around 10-20% of cells in mitosis. There are several possible approaches to achieve a certain percentage of synchronized cells in an in vitro cell culture. Basically, they can be divided in biological and chemical methods, and depending on the final use of the synchronized culture, some methods can be more appropriate than others.

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IL-37.

Importance of Monitoring Magnesium Levels in Cardiac Conditions

Dr. S. P. Dandekar Professor and Head, Department of Biochemistry, Incharge Central Clinical Biochemistry Laboratory, Seth G S Medical College & K E M Hospital, Parel, Mumbai - 400 012. Magnesium is the fourth most abundant cation in the body. It is present in more than 300 enzyme systems. A lot of interest is now being focused on the relation between magnesium and cardiovascular diseases. The following aspects regarding magnesium will be deliberated:1. The cardiac conditions under which the magnesium level determination proves useful. IL-38. 2. The clinical relevance of total and ionized magnesium levels. 3. Preanalytical, analytical and postanalytical factors to be monitored for magnesium estimation in order to obtain a clinically useful result. 4. The correlation of other electrolyte levels with magnesium. How important is it?

Tissue Zinc Levels in Carcinoma

Dr. R.Selvakumar Professor & Head, Dept Of Clinical Biochemistry, C.M.C. Hospital, Vellore - 632004 Email: [email protected] Colon cancer is among the most common human malignancies and remains a leading cause of cancer related morbidity and mortality. The transition from normal mucosa to adenoma and its subsequent progression to carcinoma are events that offer opportunities for preventive interventions. The role of zinc in carcinoma development has been the subject of debate and reports of zinc values in the biological fluids from cancer patients are often conflicting and contradictory. Hence the mechanism by which plasma zinc and tissue zinc decrease in various cancerous tissues is still obscure. The disruption of zinc homeostasis can play a significant role in the development and progression of cancer. In order to investigate the relevance of the tissue zinc and zinc related enzymes such as CuZnSOD and alkaline phosphatase activities in the development of precancerous and cancerous process in the colon, we have used well established colon cancer model in the rat. Tissue zinc levels were analyzed in the various parts of the G.I tract and other organs of normal healthy rats. There was a gradual decrease in tissue zinc concentration along the various regions of the stomach from fundus, body and antrum which was statistically significant. The highest levels of tissue zinc were found in the esophagus, testes and prostate. The DMH induced precancerous and colon carcinoma model showed that dysplasia and carcinoma were established histologically in rats at four months and six months respectively. These histological changes were directly related to a significant decrease in plasma zinc, tissue zinc, CuZnSOD and alkaline phophatase activities when compared to controls treated with saline. The mean % decrease in all the 4 parameters was greater in the carcinoma model as compared to the precancerous model. Besides the significant decrease in tissue zinc and zinc related enzymes in various parts of the G.I

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tract (stomach and small intestine) in the carcinoma model shows that transmural changes along the wall of the G.I tract could be the cause of the decrease. The above findings support the view that decrease in tissue zinc and zinc related enzymes are representative of biochemical alterations in the colonic mucosa which may reflect cellular changes that progressed to dysplasia and frank malignancy. The high incidence and mortality of colon cancer make effective prevention an important public health and economic issue. Suppression or reversal of the carcinogenic process in the colon with nonpharmacologic or pharmacologic agent is an area of considerable interest. Hence the role of the 3 agents such as Vitamin C, aspirin and zinc in DMH induced precancerous and cancerous models in rat were evaluated in this study. The results indicated that co-administration with aspirin or vitamin C in the precancerous and cancerous model maintained normal histology in the colon in 88% of rats along with a relatively lesser decrease in colonic tissue zinc levels whereas plasma zinc and IL-39.

zinc related enzymes showed no significant changes. Co-administration with zinc in the precancerous and carcinoma group prevented histological changes in the colon in all rats. The colon maintained normal histology with no significant changes in the biochemical parameters i.e plasma zinc, tissue zinc and zinc related enzymes as compared to controls. In our study on stomach cancer and colon cancer in humans, a significant decrease in tissue zinc in the cancerous tissue was observed as compared to their paired non-cancerous tissue of the same individual. In patients with prostate cancer, we observed a significant decrease in mean tissue zinc levels in prostate cancer and in BPH as compared to normal prostate from autopsy cases. Plasma zinc was significantly decreased in prostate cancer as compared to BPH and normal subjects. Urine zinc /creatinine ratio was significantly increased in prostate cancer as compared to BPH and normal subjects.

Prenatal Screening for Chromosomal Disorders

Päivi Laitinen Docent, Laboratory, Oulu University Hospital, Oulu, Finland Prenatal diagnosis has an important role in the management of pregnancies. In Western countries the current trend of family planning is toward smaller families and the average age women have babies is increasing, which increases the risk for fetal chromosomal disorders. Today women are also more aware of the frequency and the importance of congenital disorders. Modern trisomy screening combines the first trimester serum biochemical test with the thickness of the fetal nuchal translucency. The combined test improves the detection rate and reduces the false

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positive rate, compared to either biochemical or nuchal translucency testing used alone. Pregnant women and health care providers in Finland prefer early testing in order to gain more time for consideration of further procedures in case of a positive test result. The second important issue is to find a balance between testing sensitivity and invasive procedures. Down's syndrome is the primary reason for families to seek prenatal counseling. Effective first trimester screening for chromosomal abnormalities is provided by a combination of maternal age, gestational age, fetal

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nuchal translucency measurement and maternal serum free B-hCG and PAPP-A concentrations. In Northern Finland we started prenatal screening program in 2002. In 2002-2007 we have analyzed 25 000 samples for biochemical markers. In 17 000 cases also nuchal translucency was IL-40.

measured. The screen positive rate of the combined screening was 4.5% and the detection rate for Down syndrome was 84%. These results are in accordance with the published results in other screening centers.

Routine Screening of Newborns for Metabolic Disorders using Tandem Mass Spectrometry

Rohit Cariappa Chief Scientist, NeoGen Labs, Bangalore Routine high throughput newborn screening for metabolic disorders using Tandem Mass Spectrometry (MS/MS) has been carried out in developed countries for the past 15 years. Based on the screening of over 4 million babies the process has been optimized such that several hundred samples per day can be analyzed without the direct interaction of a metabolic expert in every sample. The false positive rate for the detection of over 45 metabolic disorders using licensed software and interpretation algorithms is currently less than 0.1% and the false negative rate is 0%. Tandem Mass Spectrometry (MS/MS) technology routinely screens for several classes of metabolic disorders like fatty acid oxidation, amino acid and organic acid disorders all from one small disc of dried blood in a single analytic run. Because it is much more sensitive, specific, reliable and comprehensive than traditional assays it has found widespread use in newborn screening programs around the world. It has replaced most of the oneanalysis, one-metabolite, one-disease classic screening techniques with one-analysis, multiplemetabolites, many-diseases approach. The conventional approach of first describing a disease and then noting which metabolites are increased has given way to the reverse approach which first measures a large number of metabolically important analytes from a routine blood sample collected during the newborn period. Then, if certain individual analytes or multi-analytes and analyte ratios are elevated or decreased outside of normal ranges, the software pinpoints the exact metabolic disorder. The distinct benefit of early routine screening is that the pre-symptomatic detection of a metabolic disorder allows for corresponding early treatment which increases the likelihood of excellent outcomes. Treatment for the different classes of metabolic disorders can be in the form of dietary modification or special formulas, vitamin or L-carnitine supplementation or simple avoidance of certain foods, drugs and chemicals for the affected individuals. The undiagnosed or untreated outcome for those afflicted with a metabolic disorder is very severe, ranging from physical disability, to mental retardation or sudden death. Once the symptoms of metabolic disease manifest, it is often too late for treatment to have a good outcome. Thus, newborn screening is best performed 1- 14 days after birth to have an optimal outcome.

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IL-41.

Diagnosis of Aminoacidurias by HPLC: A Few Interesting Cases

Dr. Kannan Vaidyanathan Clinical Assistant Professor, Dept of Biochemistry and Head, Metabolic Disorders Laboratory, Amrita School of Medicine, Kochi, Kerala, INDIA ­ 682 041, Ph No: 0484-2858394; Email ­ [email protected], [email protected] Objectives: Aminoacidurias are one of the oldest known classes among the various inborn errors of metabolism. Using the technique of HPLC, we wanted to detect the common aminoacidurias in our population. Methods: We have used high performance liquid chromatography (HPLC) for the detection of aminoacidurias in 350 consecutive clinically suspected cases over a period of two and a half years from November 2006 ­ April 2009. The advantages of the method include ­ 1. Speed ­ The entire analysis can be completed in 30 minutes, 2. Sensitivity ­ Sensitivity is very high, and 3. Reproducibility. Cases were suspected for aminoacidurias based on clinical presentation and were from all parts of Kerala. Age range was 0 ­ 12 years. hyper phenylalaninemia), tyrosinemia, maple syrup urine disease and nonketotic hyperglycinemia. We have detected 12 cases of maple syrup urine disease, one case of isovaleric aciduria, 2 cases of phenylketonuria, two cases of nonketotic hyperglycinemia and 5 cases of hypertyrosinemia. In addition, we have seen 56 cases with mild hyperglycinemia, and 40 cases with mild hyperalaninemia. Other cases detected include hypermethioninemia ­ 5, hyperprolinemia ­ 17, hyperhistidinemia ­ 8 and hyperlysinemia ­ 2.

Conclusions: The incidence of aminoacidurias like maple syrup urine disease is very high. Other disorders including hyperphenylalaninemia, tyrosinemia and nonketotic hyperglycinemia were also detected. Early detection of these cases is Results: We have obtained a wide range of cases possible if newborn screening is done. Therefore of which some will be discussed. These include there is increased need for newborn screening hyperphenyl alaninemia (PKU and non-PKU program in our country. IL-42.

Specific Plant-Derived Products with Relevance to Clinical Medicine

P.G Latha Tropical Botanic Garden and Research Institute, Palode, Thiruvananthapuram-695562 Medicinal plants have been a major source of therapeutics since time immemorial. They are therefore an important commodity item of international trade and commerce. Many of them are sources of clinically useful prescription drugs, being used in modern systems of medicine. Quinine extracted from the Cinchona tree (Cinchona pubescens) (Rubiaceae) and artemesin extracted from the Chinese medicinal plant,

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Artemisia annua (Asteraceae) are examples of extremely important drugs obtained from medicinal plants. The use of artemesin in the treatment of chloroquine resistant malaria particularly in Asia is well established. Atropha belladonna (Solanaceae)is the source of the CNS stimulant, Atrophine.Allium sativum (Liliaceae)"Vellulli"is the garlic which prevents arterosclerosis in blood vessels.Aloe vera

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(Liliaceae) ­ "Kattarvazha",of which the sap of the cut fleshy leaves forms Aloevera gel , used externally for burns. Cassia aungustifolia (Senna) is a stimulating laxative. Curcuma longa (Zingiberaceae)-the Turmeric or "Manjal" is clinically used against dyspepsia and flatulence, duodenal ulcers and stiffness of joints.Euphoria hirta (Euphorbiaceae)- "Nilapala" is used clinically to control amoebic bloody diarrhoea . Reserpine from Rauwolfia serpentina (Apocyanaceae) is used to reduce blood pressure.Both experimental and clinical data support use of ginger (Zingiber officinaleZingiberaceae ) for nausea , vomiting and digestive problems.Vincristine and Vinblastine, known for their anticancer property are obtained from Catharanthus roseus. Berberine from Berberis IL-43.

aristata, Coscinium fenestratum and Tinospora cordifolia are used as febrifuge and against gastritis.Xanthotoxin from Ammi majus and Heracleum candidans is used against gout and leucoderma. Podophyllotoxins from Podophyllum hexandrum are effective against cancer. Morphine and codeine from Papaver somniferum are used as sedatives and analgesics.Psoralen from Psoralea corylifolia is used against leucoderma. Hyosine and hyosyamine from Hyosyamus niger and Datura strammonium , provide sedatives and antispasmodics. In the present lecture, an overview of the specific plant derived and therapeutically significant herbal products will be presented. The important indigenous plants with significant research leads leading to drug development will also be highlighted.

Natural Products and Therapeutic Strategies: A Mechanistic Approach

Bipin Nair Professor and Dean, Amrita School of Biotechnology, Amrita University, Amritapuri, Kollam, Kerala, 690525, India. Tel: +91 0476 2896777,Email: [email protected] Natural products of plant, microbial or animal origin have for long, been used in the treatment of human diseases. As such, natural products still offer unmatched structural diversity, especially as new environmental niches are explored. Our studies have focused on attempting to understand the mechanisms involved in enhancing the rate of wound healing and the role of natural product extracts in this process. In particular, information on the role of Matrix Metalloproteinases (MMPs) in wound healing is scanty. Our investigation into IL-44. the regulation of MMPs by natural product extracts has yielded interesting information. Increasing our basic knowledge of the mechanisms involved in the process of wound healing and the strategies for pharmaceutical intervention may ultimately help clinicians to identify and proactively intervene, in an effort to prevent normal wounds from becoming chronic. This may also help to prevent the high prevalence of morbidity associated with this significant health problem.

ABC of Monoclonal Gammapathy and its Indian Scenario

T.Malati Senior Prof & Head, Department of Biochemistry, Nizam's Institute of Medical Sciences, Punjagutta, Hyderabad 500082, AP . Email: [email protected] The neoplastic proliferation of one and rarely two specific clones of b cells / plasma cells results in amplification of a large number of such clones which produce a single type of immunoglobulin

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molecule of homogeneous character, belonging to one of the five classes of immunoglobulins consisting of one type of two heavy and two light chains. Apitz (1940) for the first time used the term `paraproteins' to denote proteins produced by myeloma cells. The term now stands for `monoclonal' and is coupled with globulin, peak, spike, component etc. Multiple myeloma, arising in the marrow of bone, is a predominant plasma cell neoplasm belonging to the broad group of monoclonal gammapathy. These primary bone tumors comprise approximately 35% to 43% of total hematological malignancies. Bone pain, pathological fractures anemia, renal failure and neurological manifestations are the main symptoms associated with monoclonal Il-45.

gammapathy. The presentation will highlight 1) the methodology used for the diagnosis of monoclonal gammapathy 2. Changes in biochemical analytes occurring as a cause or effect of neoplastic proliferation of plasma cells 3) role of `m' protein, b2 microglobulin and serum c-telopeptide as tumor markers individually or in combination in diagnosis, assessing efficacy of any treatment modality (chemotherapy, radiation therapy, antiresorptive therapy) predicting early recurrence, assessing status of regression, analyzing progression of disease and finally predicting prognosis 4) the efficacy of urinary c- telopeptides in evaluating and monitoring the bone resorption process.

The Clinical Significance of Microalbuminuria

T.Malati Senior Prof & Head, Department of Biochemistry, Nizam's Institute of Medical Sciences Punjagutta, Hyderabad 500082, AP; [email protected] Microalbuminuria is associated with adverse health outcomes in adults. An alarming and steady increase of diabetics globally warrants the regular screening for microalbuminuria to identify nephropathy at an early stage during the course of disease in order to initiate timely intervention. Diabetic nephropathy and end stage renal disease (esrd) in india are emerging public health problems and therefore pose real challenge to health care professionals. Diabetic nephropathy accounts for nearly 11% deaths in patients of type 2 diabetes. Progression of nephropathy in diabetes patients is the single most common cause of end stage renal disease (esrd) world wide. An early detection and proper management of the disease is therefore essential to prevent the morbidity and mortality caused by disease. The course of diabetic nephropathy is characterized by urinary albumin excretion (uae) and glomerular filtration rate (gfr). The association of glycemic control with microalbuminuria has been established by various studies. Improving glycemic control, aggressive anti hypertensive treatment and the use

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of ace inhibitors definitely help in reducing the rate of progression of nephropathy in type1 as well as in type2 diabetic patients. Few studies support the concept that microalbuminuria is not predictive of diabetic nephropathy, but rather is an earlier point in the identification of diabetic nephropathy. The importance of measuring microalbuminuria is well established however controversies still exists regarding the type of urine specimen to be used for detecting early renal impairment in diabetic patients. The clinical significance of microalbuminuria in non diabetic population is not fully clear. The microalbuminuria is also detected in cases of essential hypertension, systemic lupus erythmatosis etc. The other factors which are associated with microalbuminuria are alcohol intake, foot ulcers and smoking. The awareness regarding the need for regular monitoring of albuminuria and proteinuria is less among health care professionals in developing countries like india. Screening for complication is particularly difficult at the primary care level due to lack of resources, time and training of health professionals.

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IL-46.

Biochemical Changes in Mountain Sickness

Savitha Nagabhushan, Thuppil Venkatesh, Vidyasagar Casikar2 E Mujica , J. Aliaga, N.Lopez, C.Smith, F. Bartholomew and M.Mongelli Traditionally mountain sickness has been regarded as a dysfunction of respiratory physiology. We have approached this problem differently based on our experience during high altitude trekking. 1. We found significant drop in the cerebral blood flow. This was absent among the Sadhus who were practising Pranayama. We looked at the influence of olfactory system on mountain sickness Albino rats whose olfactory lobes were removed were subjected to high altitude states under laboratory conditions Blood and urine samples were collected at various stages to measure biochemical parameters. Results suggested that the olfactory system regulated the pituitary function. High altitude sickness is primarily a neurological dysfunction. Olfactory system regulates physiology at high altitude through its connections with the median fore brain bundle. Hypothalamus-pituitary-Adrenal axis is influenced by the olfactory system through this pathway. 2. Andean Indians have used coca leaves (Erythroxylon coca and related species) for centuries to enhance physical performance.. . The benefits and disadvantages of using coca leaf have been a subject of many political debates. The aim of this study was to investigate the effects of chewing coca leaves on biochemical and physiological parameters. Cutaneous IL-47. microdialysis catheters were used to estimate systemic biochemical changes. We subjected 10 healthy adult males (local residents) in Cajamarca (Peru, altitude 2700 m) to a standardised exercise routine on a stationary cycle ergometer. The blood pressure, oxygen saturation (digital), pulse, VO2 max and ECG (Holter monitor) were recorded before the exercise. Cutaneous microdialysis catheters were introduced in the forearm. . The subjects were given to chew 8 g of coca leaves with a small amount of lime. They were then placed on the cycle ergometer for 20 minutes. Blood pressure, oxygen saturation, pulse, ECG and VO2 max were recorded. Pyruvate, glucose, lactate, glycerol and glutamate levels were estimated. These results suggest that coca leaves have blocked the glycolytic pathway of glucose oxidation resulting in accumulation of glucose and pyruvate. The energy requirement for exercise is being met with beta-oxidation of fatty acids. The glycerol released was also getting accumulated since its pathway for oxidation was blocked. These experimental findings suggest that chewing coca leaves is beneficial during exercise and that the effects are felt over a prolonged period of sustained physical activity. The clinical significance of these biochemical changes will be discussed.

The High Altitude Adaptation Formula

GR Zubieta-Calleja, P.E. Paulev, L Zubieta-Calleja & G Zubieta-Castillo High Altitude Pathology Institute (IPPA) & Zubieta University, La Paz, Bolivia. [email protected], www.altitudeclinic.com We have previously determined that a full adaptation to high altitude hypoxia is achieved when the hematocrit reaches a plateau corresponding to the normal level at that specific altitude. Adaptation from the hematocrit point of view, is defined as having three stages: 1) Acute, first 72 hours, where Acute mountain sickness (AMS), high altitude pulmonary edema (HAPE)

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and high altitude cerebral edema (HACE) can occur 2) Subacute, from 72 hours until the slope of increase of the hematocrit is 0, where high altitude subacute heart disease (HAHD) can occur and 3) Chronic, where the plateau of hematocrit level begins and where the normal residents achieve their optimal hematocrit. In this third stage chronic mountain sickness (CMS) subjects (polyerythrocythemic patients) present an elevated hematocrit, above that of normal individuals, for the same altitude, due to a spectrum of medical conditions originated fundamentally by cardiopulmonary deficiencies, often un-noticed at sea level. Adaptation is altitude and time dependent following the equation: IL-48.

Time Altitude For a fixed altitude the only variable that changes is Time. Hence the formula becomes a linear relation such that: Adaptation in days = Altitude in Km * 11.4. This simplification is made for easy interpretation only, as the true equation is quite complex and follows the exponential change in barometric pressure. Furthermore, for constantly changing altitudes such as in mountain climbing, this formula cannot be used in its simple form. This formula is again analyzed in an attempt to achieve a more precise focus.

Adaptation =

Do Over 200 Million Healthy Altitude Residents Really Suffer From Chronic AcidBase Disorders?

Gustavo Zubieta-Calleja (Jr), Gustavo Zubieta-Castillo (Sr), and Poul-Erik Paulev High Altitude Pathology Institute, La Paz, Bolivia. [email protected] As the oxygen tension in the inspired air (PIO2) falls with increasing altitude in normal subjects, hyperventilation ensues. This acute respiratory alkalosis is compensated by increased renal excretion of bicarbonate returning the pH back to normal. The status is called totally compensated respiratory alkalosis or chronic hypocapnia. It seems to be a contradiction that so many normal people should live their whole lives as acid-base patients. In 1865 blood gas analyses performed at 3510 m, the PaCO2 (arterial carbon dioxide tension +-SEM) = 29.4+-0.16 mmHg and pH = 7.40+-0.005. Base excess, calculated with the Van Slyke sea level equation is - 5 mM (milliMolar or mmol/L) as an average, suggesting chronic hypocapnia. Consequently, the Van Slyke sea level equation for the calculation of base excess must be adapted to altitude and should be zero. "Base excess" is misleading, since the essential variable is not a base and in reality not always an excess (or deficit). The essential concept is the titratable hydrogen ion. We have modified Siggaard84

Andersen´s term "concentration of titratable hydrogen ion" to "Titratable Hydrogen Ion concentration Difference from normal", THID now used, instead of base excess. THID is ideally determined by titration to a pH of 7.40 at a PaCO2 of 5.33 kPa (40 mmHg) at sea level, oxygen saturated and at a blood temperature of 37oC. The THID in vivo is independent of PaCO2, since any change of PaCO2 implies opposite molar changes of the bicarbonate and the non-carbonic buffer concentrations. Consequently, there is no change in THID, so the value is constant (normally equal to zero) during acute changes in PaCO2 by hyperor hypoventilation. The buffer capacity of the carbon dioxide ­ bicarbonate buffer is high, since respiratory elimination of CO2 and bicarbonate excretion of the kidneys rapidly maintains PaCO2. We have introduced a modified Van Slyke equation for high altitude (1). Since our Van Slyke equations operate with normal mean values for PaCO2 at the actual altitude, a calculation of THID will always result in normal values - that is zero.

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Reference: 1. Paulev P E, Zubieta-Calleja G R. Essentials in the diagnosis of acid-base disorders IL-49.

and their high altitude application. J Physiol and Pharmacol 56, Suppl 4, 155-170, 2005.

The Ventilatory Hyperoxic Shunt Test at 3510 m. of altitude

G Zubieta-Castillo, L. Zubieta-Calleja, and GR Zubieta-Calleja, MD High Altitude Pathology Institute(IPPA),ZubietaUniversity,La Paz, Bolivia. [email protected], www.altitudeclinic.com We use a new method to test pulmonary shunt by hyperoxia at high altitude. The subject is tested first breathing ambient air in the seated position. Using a face mask, ventilation, ETO2, ETCO2, ECG, pulse oximetry and blood pressure are measured. An arterial blood sample is taken from the radial artery. The whole procedure is repeated with the subject breathing 89% oxygen during 6 minutes. A 100 % O2 could be used, but it was not available. This technique allows for precise measurements that provide a clear picture of cardio-respiratory parameters. Furthermore it allows the possibility of observing when there is an equilibration of the PIO2 as it is observed on a t-students reports: SpO2 in % VE BTPS Ambient Air 89% O2 p= IL-50. 91.56 ± 1.45 98.15 ± 0.73 <0.0001 9.19 ± 2.3 8.26 ± 2.06 NS ETCO2 mmHg 28.05 ± 1.9 26.17 ± 2.2 <0.0001 ETO2 mmHg 66.42 ± 4.05 PaO2 mmHg 59.3 ± 5.92 PaCO2 mmHg 27.7 ± 3.72 27.25 ± 3.6 NS pH 7.43 ± 0.03 7.46 ± 0.06 NS breath by breath basis in the same wave that the ETO2 is observed. The average PIO2 was 421.7 mmHg. For 3510 m, the altitude of our Laboratory, the test performed in 14 normal subjects (mean ± SD) using This tests permits the operator to observe when the pio2 reaches the maximum level thereby making the precise observation of shunt, irrespective of uneven ventilation that can delay such equilibration of inspired gases. The intrapulmonary shunts are essential for evaluation of patients with chronic mountain sickness at high altitude.

337.59 237 ± 14.7 ± 25.8 <0.0001 <0.0001

Get Together on Biochemistry - An Experimental Teaching

Dr.W.Ebenezer William Professor & Head, Department of Biochemistry, SRM Medical College Hospital & Research Centre, Kattankulathur, Tamil Nadu ­ 603 203 E.Mail: [email protected] This is to share my experiences of experimental teaching of Biochemistry by programming a Quiz in Get-together on Biochemical topics which are minced with extra curricular activities like drawing rangoli on biochemical structures, group singing on the themes of ketogenesis, B-complex, TCA

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cycle and Hemoglobin biosynthesis along with light music songs, mimicry, skits and instrumental music by the talented first year Medical & Para medical students of various Medical Colleges of Tamil Nadu. The quiz programme had no quiz master; instead, the participating teams quizzed each other. The team answering correctly went on to question the next team and so on. This format was thoroughly enjoyed by the participants and spectators. During the rehearsal the interaction of the students with the Faculty which gives an extraordinary interest in the subject ­ Biochemistry and also during the "Get together" IL-51.

the students will experience their own leadership qualities, boldness to face the people by showing their academic interest and under hidden talents since the programme is being organized by the students, for the students and to the students. While senior Faculty of various Clinical and Para clinical departments and administrators are felicitating it motivates the students mind to acquire knowledge on medicine (Biochemistry) through the Inter ­ disciplinary approach. All these efforts are to make the "Lighter side of the Biochemistry to become a Brighter one"

The Role of Clinical Chemist in the Management of PatientRay U Department of Pathology, Royal Hobart Hospital, University of Tasmania, Australia. Clinical chemistry tests make up 75-90% of the laboratory tests request laboratories around the globe receive from the clinicians and others for the clinical management of health and diseases of individuals in the society. As the advancement in the field of medicine progresses so is the advancement in the technology related to the field of laboratory medicine. Many time consuming and complex tests are now done easily and within an acceptable time limit by the instruments. Many of us believe or try to consider that the need of clinical chemists is diminishing as the clinicians can read the tests and move accordingly for the appropriate clinical outcome. Patients in the hospital system are largely managed clinically by the junior doctors and in studies done globally found IL-52. that laboratory tests are ordered at random, very few of them understand the implications and many of them do not even know how to interpret. In a real time scenario a proactive clinical chemist addressing these issues can substantially contribute to the delivery of cost effective health service to the community by using his/her knowledge and expertise judiciously during the liaison with the clinicians responsible for the patients' management by adding or restricting the tests repertoire to help in the diagnosis, prognosis, and monitoring of the patients. If critically assessed a wise and proactive clinical chemist can take a leading role in the safe and effective delivery of the health service to the ailing subjects following the principles of evidence based practice of medicine.

Maturity Onset Diabetes in the Young (MODY)

V.Parameswaran, A.Duffield, S.Powell and J.Burgess Diabetes and Endocrinology Services, Royal Hobart Hospital, Tasmania, Australia Maturity onset diabetes of the young (MODY), characterised by lack of insulin dependence at onusually diagnosed before the age of 25, is a ge- set, absence of ketosis and is autosomal dominant netically heterogeneous form of diabetes. It is in transmission. Approximately 2% - 5% of pa86

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tients with type 2 diabetes fall into the MODY category. Six genetic forms of MODY have been recognised. MODY-3 due to hepatocyte nuclear factor 1a (HNF 1a) is the most common amongst various ethnic populations with a prevalence rate of 21% - 64% while MODY-2 caused by Glucokinase gene defect being the second most common accounts for ~14%. The varied presentation of IL-53.

MODY poses a problem in the biochemical diagnosis of this condition. Understanding of the criteria for differential diagnosis of Type 1 & 2 diabetes with the MODY type is important as a single clinical criterion may not apply to clearly delineate the presentation of this condition. The underlying pathogenesis and diagnostic criteria with examples will be presented

Post -transplant Diabetes Mellitus Following Immunosuppressive Therapy

Dr. Pragna Rao Professor and Head, Department of Biochemistry, Kamineni Institute of Medical Sciences , Narketpaly, Andhra Pradesh Diabetes mellitus is a metabolic disorder that is also a recognized complication of organ transplantation. Post-transplant diabetes mellitus (PTDM) is a form of type 2 diabetes mellitus, which is thought to develop in response to a relative insulin deficiency resulting from increased insulin resistance or impaired insulin production, or a combination of both. Transplant recipients are at a particularly high risk of developing PTDM as a consequence of factors additional to those that affect the general population, including the immunosuppressive agents used in transplant management protocols. PTDM is not a separate entity, but a symptom of an underlying metabolic disorder, which is uncovered by immunosuppression. While both tacrolimus (TAC) and ciclosporin (CsA) are known to be associated with PTDM, the mechanisms underlying this metabolic disturbance and the relative contribution of concomitant corticosteroids have been unclear There is no single, clear definition of PTDM. Depending on the definition, incidences of approximately 5-40% have been reported in patients after renal transplantation. Some investigators only include those patients using insulin treatment for a period of at least 30 days , while others also include patients taking oral glucose-lowering drugs (need for drug treatment). Several risk factors for the occurrence of PTDM have been described. These include genetic factors, race, family history of diabetes, older age, obesity, and the use of calcineurin inhibitors and steroids. Steroids disturb glucose tolerance by enhancing insulin resistance. They play a significant role in the development of PTDM. The mechanism by which calcineurin inhibitors such as cyclosporine and tacrolimus may cause impaired glucose metabolism has not been completely elucidated. CsA also has an effect on glucose metabolism. In vitro experiments have shown that CsA dosedependently inhibited insulin secretion . In the initial report comparing CsA and TAC, it was mentioned that the incidence of PTDM was comparable . The pivotal trials, however, showed significantly higher incidences with TAC . A defect in insulin synthesis as a result of an mRNA transcriptional defect was found to be dependent on duration of exposure to TAC in rats. In clinical studies, an increase in insulin resistance was suggested as an additional mechanism underlying the development of PTDM. Because both hyperinsulinemia and hyperglycemia are known to promote atherosclerosis, PTDM probably contributes to the high prevalence of death from cardiovascular disease seen in this population. Furthermore, the preponderance of infections associated with diabetes mellitus is likely to increase the risk for sepsis in the transplant

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population. PTDM is responsible for an increased incidence of graft failure ; however, to what extent this is caused by the development of diabetic nephropathy is not clear Hyperglycemia itself may upregulate the synthesis of such soluble mediators of fibrosis in the kidney as transforming growth factor-ß, which may be important in promoting IL-54.

mesangial matrix expansion and mesangial cell hyperplasia. These growth factors are likely to have a role in the development of chronic allograft nephropathy, which was the predominant cause of graft loss in the observational studies of graft outcome with PTDM.

Point-of- Care testing (POCT)

C.V.Anand Professor and Head, Dept of Biochemistry, PSG Institute of Medical Sciences and Research, Coimbatore-641004. POCT, defined as diagnostic testing at or near the site of patient care, is to facilitate rapid diagnosis and faster treatment decisions to improve patient care and reduce morbidity and mortality. POCT impacts every branch of the healthcare system. The devices should be simple to use, robust in terms of storage and usage and, produce results that are concordant with the results of the central laboratory. The results should be consistent with clinical need and, most importantly, they should be available without any delay. The POCT equipment employ different analytical principlesfor example, reflectance, lateral-flow or flowthrough immunoassays, electrochemistry, light scattering, immunoturbidimetry, spectrophotometry, fluorescence and electrical impedance, to name a few. The most frequently used equipment is the glucometer, which was introduced more than three decades ago. POCT has contributed immensely to the diagnosis of sepsis, acute coronary syndrome, stroke, GI IL-55. bleeding, kidney failure and DKA. Now a whole gamut of tests are performed using POCT equipment; cardiac markers- troponins, brain natriuretic peptide (BNP), NT-pro BNP, blood gases, plasma lipids, glycated hemoglobin etc. The major argument against POCT is that, it is carried out by personnel with no formal training in laboratory technology. Needless to say that, issues such as calibration, quality control and documentation take the back seat. Added to these are the high costs and poor comparability with central laboratory results. The advantages of POCT are reduced TAT, and plausible improved patient management. POCT has also been shown to lead to shorter hospital stay and improved health outcomes. To illustrate, POCT for heart type fatty acid binding protein (H-FABP), an early marker of acute myocardial infarction has been recently introduced, providing a significant benefit for patients at small rural hospitals.

Protein Misfolding Diseases

Dr.Usha Anand Professor, Department of Biochemistry, PSG Institute of Medical Sciences & Research, Coimbatore-641 004. Protein misfolding / conformational diseases diverse disorders which have a common feature at consist of a group of clinically and pathologically the molecular level; the presence of a protein that

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acquires a non-native, atypical conformation. Based on the causative factor, protein misfolding diseases could be broadly classified into three categories. a) Reduction in folding efficiency leading to a fall in the levels of the protein involved. Cystic fibrosis is caused by defects in the cystic fibrosis transmembrane conductance regulator (chloride channel). The misfolded protein is tagged by ubiquitin and degraded resulting in a decrease in the levels of this transmembrane protein. b) Diseases in which misfolding of a given protein results in improper trafficking. In early-onset emphysema, alpha-1antitrypsin molecules which are formed in the liver undergo polymerization. They are retained in the liver and will not reach the lungs. c) Diseases associated with aggregation of misfolded proteins. In Alzheimer's disease accumulation of a betaamyloid peptide leads to the formation of extracellular plaques. A second protein known as `tau IL-56.

protein' is responsible for neurofibrillary tangles. AD primarily affects neurons of the hippocampus and basal fore-brain leading to brain shrinkage and loss of memory and intellect. Transmissible spongiform encephalopathies: (Example: Crutzfeldt-Jakob disease) The misfolding is initiated and propagated by an abnormal protein designated as PrPsc (prion protein scrapie), a form which has extensive beta pleated structure. Huntington's disease in which the `Huntington protein' with polyglutamine repeats have been demonstrated. Amyptropic lateral sclerosis caused by mutations in the anti-oxidant enzyme superoxide-dismutase. Non-functional SOD aggregates cause this disease. With intense research in this field the number of diseases associated with protein misfolding is on the rise. A thorough understanding of the mechanisms involved would go a long way in instituting effective therapeutic measures.

Obesity ­ A Global Dilemma

V.Parameswaran, L.Blackwell, L.Diggle, J.Burgess and R.McCallum Diabetes & Endocrinology Services, Royal Hobart Hospital, Tasmania, Australia Obesity, a leading cause of morbidity and mortality, is associated with a spectrum of diseases particularly diabetes, cardiovascular disease, cancer, osteoarthritis and breathing disorders. An obese person runs a 50% increased risk of premature death than someone of normal weight. Genetic factors and also environmental factors are involved in the development of obesity. Around seven million (~ 30%) Australians are in the obese category and this trend is also reflected globally. Along with other conditions that are related to obesity, the risk of developing insulin resistance and diabetes is also increasing rapidly. Research in finding treatment and effective management is being constantly sought and trialled to try and mitigate the clinical effects of obesity. Common approaches include diet, exercise and lifestyle modifications. Recent approaches have focussed on newer drugs as also the additional option of surgery in the management of obesity in those where it is refractory by other means. The need for appropriate screening to reduce the risk of developing diabetes is important. Some of these strategies along with dietary and lifestyle change programs, new diabetes medical treatments, lap-band surgery will be presented and discussed.

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IL-57.

Scholarly Communication: Writing a Research Paper for a Peer-Reviewed Publication

R S Jayasomu National Institute of Science Communication and Information Resources (CSIR), New Delhi Communicating science is as important as doing science. Knowledge is perhaps the only commodity apart from love and peace that grows when shared. Science grew effectively only after it was communicated, shared, and discussed upon. Science writing is not storey writing. It has set rules and regulations, format and style depending on what you write about, and to whom you address. Broadly, science writing can be classified into popular science writing, writing for encyclopedia, and research communication. Communicating research needs to be focused as it would address specific audience, the scholars. Conventionally, basic units of a research publication include Introduction, Materials and Methods, Results, Discussion, Conclusion, Acknowledgement and References. Data are given either in tables or expressed graphically through illustrations. Abstract and keywords are given in a separate sheet, prefIL-58 erably in the beginning following the covering letter and declaration. Generally, first page of a manuscript should have the Title, name(s) of author(s), their affiliations, contact address of author for correspondence, abbreviations, if any, and a short title. Tables and illustrations are given separately after references. Care should be taken that the table data are not repeated as illustrations. Legends or captions of illustrations are to be typed separately in a page. Once the data are ready after your experiments, it is necessary to find an appropriate journal for communicating the results of your research work. Papers published in non-core journals seldom get their due citation. In the present paper, I intend to share my knowledge gained during the last two decades in this profession of science editing on how to write a research paper

Urine Albumin: Standardisation and Reporting Issues.

H. D. Martin Gribbles Pathology, 1 Goodwood Rd, Wayville, SA 5034, Australia

Urine albumin is well established as an early marker for nephropathy in diabetic subjects. More recently it has also been identified as an important prognostic marker in hypertension and cardiovascular disease. Despite the literature supporting clinical utility there is evidence that clinicians do not often request urine albumin, particularly in their non-diabetic patients. Furthermore, when they do make the request, they find interpretation confusing and adherence to guidelines is poor. Currently there is no reference system for the

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measurement of urine albumin: in particular the measurand is not defined, there is no reference material and although the manufacturers of the majority of methods in routine laboratory use the same calibrator source, it is a serum based material and individual preparations are not standardised. Despite this, the analytical variation represents a small proportion of the total error in urine albumin measurement. It is the pre-analytical phase which contributes the major variation. Albumin excretion rate (AER) and albumin crea-

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tinine ratios (ACR) vary substantially with collection type and patient preparation. Daytime collections yield significantly higher results than nightime collections. Exercise, stress, posture, infection, hydration and blood contamination can contribute to bias in both AER and ACR. Creatinine variation will contribute significantly to variation in ACR but reference ranges are not age/ gender/race specific. Decision points for urine albu-

min, AER and ACR in common use vary widely and none has been clinically validated for the particular method. The NKDEP/IFCC has a urine albumin working group that is collaborating with other professional groups to work towards standardisation; in the meantime individual laboratories can play a crucial role in educating health care providers.

IL-59

Molecular Mechanisms of Angiogenesis in Heart and Lung

Hari S. Sharma Angiogenesis and Tissue Remodeling Group, VUMC, Amsterdam and Erasmus MC, Rotterdam, The Netherlands Right ventricular hypertrophy and failure are common features in patients with tetraogy of Fallot (TF). Myocardial angiogenic gene expression profile was examined in patients with right ventricular hypertrophy and failure in order to assess myocardial vascularisation. DNA microarray analysis on right ventricular biopsies from TF-patients operated for primary corrective surgery (TF-1, n=8, mean age 0.5 year) and age matched controls was validated by Northern hybridization and RT-PCR. Employing immunohistochemistry and video image analysis expression of vascular endothelial growth factor (VEGF), vascular density (by a-SMA and CD31 staining) and myocyte cross sectional area (Gomori's reticuline staining) were assessed in TF-1 and adult patients (TF-2, n=8, mean age 30 years) who underwent surgery for pulmonary regurgitation and compared the data with age matched controls. DNA microarray analysis revealed altered expression pattern for several genes including angiogenic factors, VEGF, flt-1, flk-1 angiopoietin-2, FGF-2, FGF-R1, PDGF-A, whereas, flt-4, Tie, TGF-b, TGF-b3R showed decreased expression in TF patients. Northern blot analysis verified VEGF expression whereas, flk1 expression remained unaltered in both TF-1 and TF-2 patients. VEGF staining in cardiomyocytes was increased in TF-1 as compared to TF-2. Video image analysis revealed enhanced vascular density with unchanged wall thickness and enlarged myocyte cross sectional area in both TF-1 and TF2 as compared to respective age matched controls. Right ventricular hypertrophy is associated with an altered gene profile for a number of genes where the VEGF/VEGF-R system plays an important role in enhanced myocardial angiogenesis with limited vascular remodeling. In another set of experiments Mechanisms of angiogenesis were studied using human Cell culture based in vitro models. We have shown that different growth factors and cytokines result in differential gene expression and secretion of various proinflammatory cytokines and vascular endothelial growth factor (VEGF), an angiogenic molecule in cultured human ASM cells. To assess the role of airway smooth muscle (ASM) in bronchial angiogenesis and remodeling, we investigated the production of VEGF in ASM cells in relation to mediators of asthma, such as, IL-1b, TNF-a, TGF-b, ANG II and ET-1. Time dependent release of VEGF protein in the conditioned medium was observed which in its turn induced proliferation and growth of pulmonary artery endothelial cells. We further investigated the effects of nitric oxide (NO) pathway on the pro91

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inflammatory cytokine; Interleukin-1b (IL-1b) induced expression and secretion of VEGF and PlGF. Serum deprived (for 48h) ASM cells were stimulated with IL-1b (5 ng/ml), IL-1b + Nw-nitroL-arginine methyl ester (L-NAME), IL-1b + Larginin and IL-1b + L-NAME + L-arginine for 4 and 24 h. NO synthase inhibitor (L-NAME) was used 1h prior to IL-1 incubation in all experiments. IL-1b induced expression (1.8 fold vs control) of VEGF mRNA (quantitative RT-PCR) was attenuated by L-NAME and augmented by Larginine. L-NAME inhibited the secretion of IL-60

VEGF and PlGF in conditioned media of IL-1b treated PASMC at 4 and 24 h, respectively. By restoring NO pathway (L-arginine treatment) in L-NAME treated cells led to elevated (2.2 fold) expression of VEGF. Taken together, our findings suggest that a cytokine cascade involving mainly IL-6, IL-8 and VEGF operates in hyper contractile human ASM cells where NO pathway may modulate VEGF signaling during airway inflammation and subsequently contributing to bronchial angiogenesis and airway remodeling in patients with asthma and COPD.

The Pro-Inflammatory Role of Extracellular RNA in Arteriogenesis

Elisabeth Deindl Walter-Brendel-Center for Experimental Medicine ; Ludwig-Maximilians-University Marchioninistr. 27 D-81377,Munich,Germany The growth of pre-existing arteriolar anastomoses to large conductance arteries (arteriogenesis) is the most important tissue-, organ-, and often life saving process after arterial occlusions. It can be so efficient that occlusions of coronary, peripheral, renal, and cerebral arteries may occur without tissue loss and even without loss of function. The initial trigger for the growth of those natural bypasses is fluid shear stress activating the endothelium surface, thereby leading to modulation of processes controlling the recruitment of circulating cells. The inflammatory aspect of this activation is apparent by the increased expression of ICAM-1 as well as of MCP-1, a chemoattractant protein responsible for the adhesion and migration of monocytes to the endothelium. Recently, extracellular RNA was described to act as "danger signal" and trigger immune response. In order to investigate the function of extracellular RNA in an animal model of arteriogenesis, the right femoral artery of mice was ligated and the left was sham operated. Arterial growth was measured via Laser Doppler Imaging (LDI) and quantified as relative perfusion

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recovery (right/left) before and after surgery until day 7. Treatment with RNase, but not with DNase, immediately before occlusion of the artery until day 7 significantly reduced the extent of arteriogenesis (perfusion recovery in untreated or DNase treated mice: 83%, in RNase treated mice 44%), suggesting that extracellular RNA plays a role in the process of arteriogenesis. Accordingly, in vitro studies confirmed that RNA acts as a chemoattractant for monocytes. In an endothelial cell monolayer the migration of monocytes was increased by RNA to nearly the same extent as by MCP-1. In addition, there was an increase in adhesion of monocytes to microvascular endothelial cells by RNA, but not by DNA. The expression of ICAM-1 on endothelial cells was increased by RNA and abolished in the presence of RNase. Furthermore, RNA led to exocytosis of pro-inflammatory factors like P-Selektin from Weibel-Palade bodies. In summary, our data indicate that extracellular RNA promotes the process of arteriogenesis by immune-modulating effects.

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Pharmacogenomics: One Size Does Not Fit All

Dr. Amit Chakrabarti. Professor of Pharmacology, Sikkim Manipal Institute of Medical Sciences, Gangtok, Sikkim 737 102, India Pharmacogenomics is the study of how an individual's genetic environment influences response of drugs. It is thus an intersection of pharmacology and genetics. Although the knowledge that genes may influence action of drugs was present since the 1930s, major advances occurred only in the last couple of decades. Fundamentally, pharmacogenomics conveys the message that no two persons are similar with respect to effect of drugs, because everyone has a different genetic milieu. This essentially promises that one day extremely safe drugs could be tailored considering a person's genetic constitution. Therefore, potential benefits of pharmacogenomics include 1) synthesis of more disease specific medicines, with less potential of damaging healthy cells; 2) prescription of best drugs for a particular patient considering genetic constitution; 3) more appropriate dose titration of medicines; 4) early screening for genetic diseases; 5) safer and cost-effective vaccines; 6) simpler drug development process identifying subjects IL-62 with a particular genetic make up for a particular drug; and 7) overall reduction in health care costs. As of now pharmacogenomics is in limited clinical use. Most prominent use is for identification of variants of the cytochrome P450 enzyme system. Before clinical trials investigators can detect if the trial participant has a different form of P450 enzyme, which may reduce drug metabolism and increase chance of adverse events. Pharmacogenomics is in its infancy and has a number of barriers to overcome before becoming truly applicable in everyday clinical practice. Some of the barriers to successful clinical translation are 1) large number of gene variations affecting drug action; 2) drug choices are not sufficient to account for all variations; 3) developing a number of drugs to fit different genetic make ups is not a costeffective drug development option; 4) drug prescription according to pharmacogenetic profile will be a complex process involving further diagnostic tests and intensive training for treatment providers.

Lead Not Safe in Environment

Dr. Krishnajyoti Goswami Director, National Referral Centre for Lead Poisoning in India, West Bengal (NRCLPI, WB); President, Association of Clinical Biochemists of India (ACBI); Professor of Biochemistry, Ramakrishna Mission Seva Pratishthan; Vivekananda Institute of Medical Sciences; 99 Sarat Bose Road, KOLKATA - 700 026. Email: [email protected] & [email protected]

Lead is one of the toxic heavy metals, which is environmental as well as health hazard, affecting globally considered as the number one people of all age groups. It has deleterious effect

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on all organs in the human body. Lead poisoning is widely recognised as a major public health problem all over the world. Every moment, we are building a stock of Lead in our body. Lead from our house paints, drinking water, food staffs, glossy cosmetics accumulate in our body creating a stock of unwanted toxic chemical ­ Lead Poisoning, a "Silent Epidemic". The human body cannot process and expel lead. Sustained and close contact can cause serious damage to developing central nervous system and brain that leads to a child performing poorly in exams or having short attention spans. Adults who are exposed to lead poisoning many find it difficult to concentrate or remember things and feel pain in muscles and joints. Children and women are found most vulnerable to lead poisoning (also known as saturnism, plumbism, or painter's colic). Lead may cause irreversible neurological damage as well as renal disease, cardiovascular effects, and reproductive toxicity. IL-63

Lead level in blood exceeding 10 ug/dl (recently 2.0 ug/dl) is considered as unsafe, but studies indicate that over 60% of children in India may have more than this level in their blood. Unlike other health hazards, lead poisoning is 100% preventable by early diagnosis and prevention of exposure by awareness. Paint manufacturers have the capability and technology to phase out lead from their products. But costs being an issue, companies complain that making lead free products makes the product expensive, and lowers their market share. It is therefore, even more important to have mandatory standards for all branded companies globally. The environmental regulation should not be confined to production but extended to products we use. The awesome effect of the "silent epidemic" needs to be understood and propagated to the mass for their awareness and effective prevention - thus "help yourself and others too".

Developments in the Biochemical Assessment of Renal Disease

David Torrens Accurex Biomedical/ Genzyme Diagnostics interference and are introducing calibration traceable to isotope dilution mass spectrometry. Equations have been developed to convert creatinine results into eGFR using factors known to affect creatinine measurement. The four parameter MDRD equation includes age, sex and race; although the only race options are Caucasian and Afro-Caribbean. There is now a more recent

Chronic kidney disease is increasing throughout the world, linked to an ageing population and an increase in type 2 diabetes. Serum creatinine measurement based on the Jaffe reaction has been central to the assessment of renal function for decades, despite a growing understanding of interferences with the method. Manufacturers have made attempts to reduce bilirubin and protein

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CKD-EPI equation and a new Schwartz equation only applicable to patients over 18 years old and for paediatric use. makes assumptions about a standardised body type. The IFCC Scientific Division and NKDEP Laboratory Working Group called for more specific creatinine methods to be adopted, and the introduction of enzymatic creatinine methods has effectively removed the major interferences seen with the chemical Jaffe reaction. The improved accuracy of enzymatic creatinine has advantages in a number of patient groups. In particular, for children and neonates where the correction factors for protein interference are inappropriate, enzymatic creatinine measurements are required. Also, eGFR is unreliable when greater than 60 mL/ min/1.73m2 and a single creatinine measurement is insensitive to moderate reductions in GFR. Cystatin C measurement overcomes many of these problems. It is produced consistently and is freely filtered at the glomerulus, not affected by age, gender, muscle mass, nutritional status or malignancy. Its use has been validated in paediatrics, the elderly and in those with diabetes and coronary heart disease. The potential for detecting earlier stages of renal disease that may

However, there are fundamental problems with be more amenable to treatment is of particular creatinine measurements. The MDRD formula is interest. IL-64

ISO 15189 : 2007

Dr. Elizabeth A. Frank Labotatory medicine and the clinical lab have oorder to give formal recognition that the laboraundergone many a changes in the last decade. tory is competent to carry out examinations based Clinical Labs now play a pivotal role in diagnosis on ISO 15189:2007. Particular requiremment for and management of disease. From being small quality and competence.ISO 15189 will be disinsignificant part of medicine the labs have cussed in detailed with the view to help labs to emerged into being sophisticated and highly au- undersatand the standard and prepare the labs for tomated aspiring to support the clinician. the need accreditation. for quality in reporting and manageement is now the need of the hour. Though India lacks regulation that makes accreditation mandatory. It is important for the Laboratories to recognize the need for accreditation and the value it brings. Accreditation is a peer-review process by which an authoritative body ensures that laboratories meet explicit quality management criteria in

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ABSTRACTS OF PRESENTATIONS

Abstract No: 1.1

A Study on Free and Total Prostate Specific Antigen in Benign Prostatic Hyperplasia (BPH) and Carcinoma Prostate

Dr. R. Lalitha Associate Professor Biochemistry, Stanley Medical College, Chennai ­ 600001 Objective: The molecular forms of Prostate Specific Antigen (PSA) are Free PSA (unbound) and bound forms ­ PSA á (antichymotrypsin) and PSA á 2 (macroglobulin). Studies established that, in BPH more free form is present than CA Prostate. This study aimed to evaluate free and total PSA in BPH and CA prostate and the ratio between free PSA to total PSA (which is % Free PSA) in discriminating between these two diseases. Free PSA and total PSA are correlated with, prostatic volume, total acid phosphatase, prostatic acid phosphatase, total PSA density and free PSA density. Methods: The study was carried out on 79 patients of age 50 and above. They constituted 40 normal, 24 BPH and 15 CA Prostate patients. Serum total PSA and free PSA were estimated by a microplate Enzyme immunometric assay (EIA). Abstract No: 1.2 Results: The two diseases were differentiated at cut off level of 6 ng/ml (sensitivity 42%, specificity 88%) of total PSA, and 0.5 ng/ml of free PSA (sensitivity 58%, specificity 88%). In, % free PSA. A cut off level of 0.1 is established between controls and prostate diseases, but a suitable cut off for the 2 diseases was not established. Conclusion: Free PSA, Total PSA, and % free PSA increased in prostate cancer, but not in controls. On contrary to the previous studies, there was no difference in any of the three entities, namely, total PSA, free PSA and % free PSA, between the two diseases. Among the biochemical parameters evaluated, total and prostatic acid phosphatases are better indicators in differentiating between BPH and CA Prostate in patients with increased prostatic volume.

Evaluation of Enzyme Markers in Carcinoma Cervix.

Dr.P.B.Desai and Dr.Chetana.K Department of Biochemistry, KLE University's J.N.Medical College.Belgaum.India, Email: [email protected] Objective: Cervical cancer is the second most common cancer in women worldwide. In India also it accounts for a quarter of the global burden of cervical cancer. As there are very few studies on enzyme markers in cervical cancer the present

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study has been undertaken to assess the reliability of some of the enzyme markers namely, serum adenosine deaminase (ADA) and gamma glutamyl transferase (GGT) which are inexpensive, analysed by easy methods and may be used as supportive

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parameters for diagnostic purpose and may add The mean serum Gamma glutamyl transferase further for prognostic information. level was 19.61 ± 5.80 U/L in controls whereas in Methods: The present study comprised of 40 healthy females in the age group of 35-65 years as controls and 40 clinically and histopathologically confirmed patients of cervical cancer of the same age group. Serum ADA was estimated by Galanti and Giusti Method and Serum GGT by Carboxy substrate method. Results: In the present study, mean serum adenosine deaminase level in controls was 20.81 ± 5.97 U/L while in the patients of cervical cancer 56.45 ± 19 U/L. A highly significant increase in serum ADA was observed in cervical cancer patients as compared to controls (p value < 0.001). Abstract No: 1.3 cervical cancer patients 40.96 ± 8.45 U/L. A significant increase in GGT was observed in cervical cancer patients when compared to controls (p-value < 0.001) Conclusions: Present study suggests that serum adenosine deaminase and gamma glutamyl transferase may be used as supportive parameters for diagnostic purpose and may add further for prognostic information. These biochemical parameters are inexpensive, rapid and can be easily analysed in smaller laboratories which have not exposed to any sophisticated technology.

Serum Fluoride and Sialic Acid Levels in Osteosarcoma

R. Sandhu, Z. S. Kundu, S. Kharb and Harbans Lal Postgraduate Institute of Medical Sciences, Rohtak (Haryana), Email: [email protected] Objective: Osteosarcoma is a rare malignant bone tumor, most commonly occurring in children and young adults, presenting with painful swelling. Various etiological factors include ionizing radiation, family history of bone disorders and cancer, chemicals such as fluoride, beryllium and vinyl chloride, and viruses. Glycoproteins and glycosamino-glycans are integral part of bone and prolonged exposure to fluoride, for long duration, has been shown to cause degradation of collagen and ground substance in bones. Methods: The present study was therefore planned to analyze serum fluoride, sialic acid, calcium, phosphorus, and alkaline phosphatase in 25 patients of osteosarcoma, and age and sex ­ matched patients with bone-forming tumors other than osteosarcoma, and a group of patients with musculo-skeletal pain as controls. Fluoride was estimated by ISE, and sialic acid by Warren's method. Mean serum fluoride concentration was found to be significantly higher in patients with osteosarcoma as compared to the other two groups. Results: Mean serum fluoride concentration in patients with other bone-forming tumors was nearly 50% of the group of osteosarcoma, however, it was significantly higher when compared with group I. serum sialic acid concentration was found to be significantly raised in patients with osteosarcoma as well as in group with other boneforming tumors as compared to the controls. Conclusion: Results suggest a role of fluoride in the disease.

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Abstract No: 1.4

Diagnosis of Breast Cancer by the Determination of Plasma Maspin Level

Karabi Ganguly (Bhattacharjee), G.V.Girish and Asru K. Sinha Sinha Institute of Medical Science and Technology, 288 Kendua Main Road, Baishnabhghata, Garia, Kolkata-700084, E mail- [email protected] Objective: The occurrence of breast cancer in women is one of the most common neoplastic conditions. Unfortunately at present the diagnosis of breast cancer by biopsy remains only option for the confirmation of the condition. We investigated the possibility of the use of the determination of plasma maspin level for the diagnosis of cancer. Methods: Maspin (a mammary serpine inhibitor), an anti breast cancer protein produced abundantly in the normal mammary epithelial cells but not in malignant breast tissue. Maspin was assayed by enzyme linked immunosorbent assay (ELISA). A total of 35 female breast cancer patients (between the ages of 45-65) participated in the study. An equal number of age matched female volunteers also participated in the study. None of the cancer patients at presentation underwent any therapy for the condition. The staging were T-1=6, T2=8,T3=9, N0=5, N1-N3= >, stage I+II=30, Stage III=5,M0=0. Maspin was assayed in the collected blood sample as soon as practicable. Results: The plasma maspin level was 4.67±0.04nM in normal volunteers (n=35). In contrast basal plasma level of maspin was 0.95±0.01nM (n=35, p<0.0001). Furthermore decrease of plasma maspin level was inversely related to the increase of malignancy grade of the condition. Conclusion: Plasma maspin level could be a useful marker for the diagnostic test for human breast cancer.

Abstract No: 1.5

Evaluation of Serum Tumor Markers and Fertility Hormones in Postmenopausal Women with Ovarian Cancer

L. Vishnu Prasad1 and Dr.R.Arivazagan2 1 Sono Diagnostics, Chennai, 2Department of Biochemistry, Cancer Institute, Chennai Objective: To observe the levels of fertility hormones along with tumor markers in ovarian cancer subjects compared with age matched controls. Materials: In this study we analysed serum samples for CA125, Free beta HCG, AFP, FSH,

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LH Prolactin and Testosterone assays in 88 Female subjects, Who were attending the oncology clinic in Chennai .All the subjects were Post Menopausal women age 40 yrs or more. Blood samples were collected from both study group with out any treatment and 20 healthy age matched subjects.

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Method: All the test were analysed. by Testosterone, Free beta HCG, AFP) is more than Chemiluminescence (CLIA) technique using 0.3814 which is not statistically significant. Immulite 1000 ­ siemens analyzer as per Conclusion: From the above data it was found that manufacturer protocol. 58 % of ovarian cancer patients have elevated Result: We have observed that out of 88 ovarian CA125. FSH was in the higher normal range in cancer subjects 51 had elevated CA125 levels. Out 52% of CA125 elevated subjects than in normal of 51 subjects who had elevated CA125 levels. subjects and in CA125 level within normal limit Mean is 996 u/l (P=0.0162) and FSH. Mean is of study group. Therefore the combination of CA 86.9 u/l (p=0.0049). The CA125 level within 125 and FSH may be a better biomarker for normal limit of Ovarian cancer subjects was not ovarian cancer patient before and on Followup statistically significant while the mean is 16.3 u/l. treatment to monitor the tumor burden and The p value for other parameters (LH, Prolactin, treatment response. Abstract No: 1.6

Serum sialic acid forms as tumors head, neck, thyroid & lung tumors.

Mahendra R. Pakhale , Dr.Murhar and Rajesh Rajrkar Dept. of Biochemistry, Yavatmal (M.S) Objective: Serum sialic acid levels are significantly elevated in thyroid tumors, head & neck, lung tumors. Such elevation correlate with tumor progression & metastasis. Generally serum lipid bound sialic acid levels are elevated in cancer patients. Methods As compared to healthy subjects & patients with chronic non-tumors diseases. Patients with metastatic disease have higher values than ones with localized tumors. Serum sialic acid levels are often elevated in patients with lung cancer. Patients in clinical remission following chemotherapy also show elevated mean serum sialic acid levels. Sialic acid levelsare also increased in nonmalignant growth of breast tissue during pregnancy & lactation. Despite the low specification of serum sialic acids as tumor markers their levels seem to be related to clinical course of disease & could be useful in the monitoring of tumor progression. Results:Total sialic acid levels were also found to be increased in serums from lung cancer patients as compared to the healthy controls. in patients with metastases these levels are further increased. As a rile serum lipid bound sialic acid is elevated in serums from patients with lung cancer. Lipid bound sialic acid serum changes could be correlated with the extent of disease.In some of the studies no statistically significant difference between patients with chronic non tumor diseases & cancer patients could be established. However, lipid bound sialic acid levels are significantly higher in patients with metastatic disease compared to ones with loclized timor. Serum lipid bound sialic acid levels hanged during intensive chemotherapy almost parallel with the clinical course, & hence lipid bound sialic acid can be of

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potential use for following the course of chemotherapy. Conclusion: Combined measurements of total sialic acid & lipidbound sialic acid in serum from patients with lung cancer could be more discriminatory than values of either of these markers. Combination of these markers might be Abstract No: 1.7

useful for differentiation between benign & malignant conditions & also for diagnosis of metaststic lung cancer. Comparison between total sialic acid & lipid bound sialic acid values could also discriminate between different tumor types or between patients with & without metastases.

Matrix Metalloproteinases and Their Inhibitors: Correlation with Invasion and Metastasis in Oral Squamous Cell Carcinoma

Ragini D. Singh1, N Haridas2, Shilin N. Shukla1, Jayendra B.Patel1, Bhairavi N. Vajaria1 and Prabhudas S. Patel1 1 Biochemistry Research Division, The Gujarat Cancer & Research Institute, Asarwa, Ahmedabad - 380 016, Gujarat, 2Pramukhswami Medical College, Karamsad, Anand, Gujarat,Email: [email protected]

Objective: Matrix metalloproteinases (MMPs) malignant tissues as compared to adjacent normal have been implicated in the invasion and tissues. A significant positive correlation was metastasis of various malignancies. demonstrated between levels of proMMP-9 and active MMP-9 with differentiation, stage and Methods: The study evaluated a comprehensive infiltration. ProMMP-2 and active MMP-2 profile of MMP-2 and MMP-9 and their main exhibited significant correlation with inhibitors, TIMP-2 and TIMP-1, respectively in differentiation and lymph node involvement. The 50 controls and 75 patients with primary oral SCC. multivariate analysis of ELISA results revealed a Blood samples from controls and patients as well as malignant and adjacent normal tissues from the significant positive correlation between MMP-2, patients were collected. The study examined pro, TIMP-1 and TIMP-2 levels with lymph node active and total forms of MMP-2 and MMP-9 involvement, stage and differentiation. using zymography. ELISA and RT-PCR were carried out to evaluate protein levels and m-RNA expression respectively for the MMPs and TIMPs. Results: Significantly higher plasma pro, active and total MMP-2, MMP-9 as well as TIMP-1 and TIMP-2 levels were found in oral cancer patients as compared to the controls. mRNA expression of the MMPs and TIMPs was significantly higher in

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Conclusion: The ROC curve analysis showed that the levels of MMPs and TIMPs have significant discriminatory efficacy to discriminate between controls and patients. The results indicate that MMP-2, MMP-9, TIMP-1 and TIMP-2 have significant clinical usefulness for oral cancer patients. Zymographic analysis is a simple, cost effective, rapid and sensitive alternative assay.

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Abstract No: 1.8

A Study of Oxidative Stress in Cancer Patient before and after Supplementation of Vitamin C and E

Dr. Pranami Bordoloi , Dr. Bikash Kr. Chaudhury, Dr. Chandan Kr. Nath and Dr. Dipali Das Department of Biochemistry, Gauhati Medical College and Hospital, Guwahati ­ 781032 Assam. Email: drpranamibordoloi @ gmail.com Objective: Cancer is the second leading cause of death behind heart disease. Increased lipid peroxidation is due to the altered ratio between free radicals and antioxidant status. The study was carried out to analyze and evaluate the relationship between antioxidant vitamin C & E supplementation and its effect on the lipid peroxidation in the cancer and normal population. Methods: We estimated Malondialdehyde, lipid peroxidation marker colorimetrically by Thiobarbituric acids Reactive Substances (TBARS) and uric acid is by Uricase method. Statistical analysis is done by Student`t' test. MDA Abstract No: 1.9 values in both the study groups decreased significantly after 7 days of vitamin supplementation (p<0.001) Results: Uric acid values in both the studied group increased significantly after Vitamin Supplementation for 7 days (p<0.001). Conclusion:The study suggested that MDA value reduced in cancer patients within one week of antioxidant vitamin supplementation but for reduction of uric acid levels longer period of vitamin supplementation is required in both the population.

Role of Serum Enzymes in Oral Cancer.

Dr P.B Desai and Dr. Shikha Sharma Department of Biochemistry, J.N Medical College, Belgaum, Karnataka,Email:[email protected]

Introduction: Oral cancer is the most common type of Head and Neck cancer and is a major health problem in developing countries. The survival index is small (50%) as compared to the progress in the diagnosis and treatment of other tumours. Thus there is a need for the improvement in detection of early stages as treatment is more effective and morbidity minimal. A majority of the initial alterations in these lesions are not readily recognizable on clinical or histopathological

examination. Therefore a number of biochemical markers have been studied to evaluate the malignancy. In view of this, the present study is undertaken to provide some promising biomarkers namely Adenosine deaminase (ADA) and 5'Nucleotidase (5'-NT) which are inexpensive, accurate and identifiable by easy methods of detection and therefore may be of some diagnostic and prognostic significance.

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Methods: A total of 20 clinically and histopathologically confirmed male patients of oral cancer in different stages were included in the study. There were in the age group of 35-65 years. Normal subjects were taken as controls. 5ml of blood was drawn and ADA was estimated by method of Galanti and Giusti and 5'-NT by method of Campbell. Results: The levels of ADA and 5'-NT were significantly elevated in oral cancer patients in comparison to controls. An increase in these Abstract No: 1.10

biomarkers was also seen when interstage comparison was done. Conclusion: The rise in ADA and 5'-NT activity in cancer patients occurs due to enzyme leakage from the primary cancerous or metastatic tissue. The rise in activity in relation to the stage further suggest that increase in tumour mass may be directly responsible for the increased levels of the enzyme. Thus these markers are of diagnostic and prognostic significance in case of Oral Cancer.

Biochemical and Hematological Markers in Patients with Chromosome 22q11.2 Deletion and Conotruncal Defects.

Alka Anilkumar,1 M.V.Thampi,1 R.Krishna Kumar,2 K.Mahesh,2 Sheela Nampoothiri,3 Sindu T.K,4 and D. M. Vasudevan.5 1 Dept. of Human Cytogenetics, 2Dept. of Pediatric Cardiology, 3Dept. Pediatric Genetics, 4Dept. of Biostatistics, 5Ex-Dean, Faculty of Medicine, Amrita Institute of Medical Sciences and Research Centre, Elamakkara, Cochin ­ 682026, Tel.0484-4008194, Fax:0484-2802020, Email: [email protected] Background: The incidence of 22q11 deletion syndrome is found to be 1 in 4000 live births. Gene dosage reduction like heterozygous deletion of chromosome 22q11.2 region leads to 22q11 deletion syndrome. Phenotypic variability is one of cardinal feature of this syndrome. Cardiac mainly conotruncal defects, hypoparathyroidism, hypocalcaemia and typical dysmorphic features are some well-described associations. Only limited data is available regarding abnormal thyroid functions and hematological manifestations in 22q11DS. Method: Retrospective study carried out on 168 children (d"2 years) with conotruncal malformations. 22q11 deletion was analysed by Fluorescence in situ hybridization technique. Total Serum Calcium level, Serum Thyroid stimulating hormone (TSH), Mean Platelet volume and platelet count were also studied.

Results: 19% of patients were hemizygous for chromosome 22q11.2 region. A higher mean platelet volume (MPV) about 10.45fL was found in deleted patient group as compared to 7.6fL in the non-deleted patients (p <0.001). A negative Objective: To identify biochemical and correlation obtained between MPV and platelet hematological markers of 22q11.2 deletion count. None of the deleted patients had syndrome. hypothyroidism. However significant association

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was found between deletion and calcium level (p microdeletion. Abnormalities of TSH level were <0.001). not found to be a feature of 22q11DS. Elevation of MPV may be due to deletion of the platelet Conclusion: Mean platelet volume, platelet count glycoprotein gene GP1BB besides other candidate and calcium levels can be used as laboratory genes like TUPLE1, TBX1 and UFDIL located on markers for clinical identification of 22q11 the chromosome 22q11.2 region. Abstract No: 1.11

Enhanced 5HT2A Receptors in the Cerebral Cortex of Hypoxia Induced Neonatal Rats: Effect of Glucose and Oxygen Supplementation

Anju T. R, Pretty Mary Abhraham and C. S. Paulose Molecular Neurobiology and Cell Biology Unit, Centre for Neuroscience, Department of Biotechnology, Cochin University of Science and Technology, Cochin- 682 022, Kerala, India. [email protected] Objective: Hypoxia at birth causes changes in brain neurotransmitters depending on its severity and durations. Serotonin (5-hydroxytryptamine; 5-HT) is one of the major neurotransmitters participating in the development of hypoxiainduced pulmonary hypertension. Ventilatory long-term facilitation during hypoxia requires 5HT2 receptors. We investigated the 5HT2A receptor functional regulation in the cerebral cortex of hypoxia induced neonatal rats and the role of glucose and oxygen supplementation in regulating the receptor alterations due to hypoxia. Methods: Wistar neonatal rats of 4 days old used for the experiments were grouped into five as follows: (i) Control (C); (ii) Hypoxia (Hx); (iii) Hypoxic neonatal rats injected with 10% dextrose (500mg/ Kg body wt) intra-peritoneally (i.p.) (Hx+G). (iv) Hypoxic neonatal rats injected with 10% dextrose (500mg/ Kg body wt) i.p. and treated with 100% oxygen (Hx+G+O); (v) Hypoxic neonatal rats treated with 100% oxygen (Hx+O). 5HT2A receptor was assayed according to the procedure of Uzbekov et al, 1979. Results: Under hypoxic condition neonatal rats showed a significant increase in receptor number (B max) with a decreased affinity (p<0.001). Resuscitation with glucose (Hx + G) and glucose along with oxygen (Hx + G + O) showed a significant reversal to near control. Hypoxic neonatal rats treated with 100% oxygen (Hx + O) showed a significant increase in Bmax (p<0.001) compared to control. Real Time PCR analysis also showed a significant increase (p<0.001) in the gene expression of 5HT2A receptors in hypoxic neonates. The gene expression showed a reversal to near control in Hx + G and Hx + G + O. Pulmonary hypertension during hypoxia can be attributed to the increase in 5HT receptors in the cortex. Glucose alone and glucose + oxygen supplementation reversed the altered receptors in the developing brain of hypoxic neonatal rats. Conclusion: This has clinical significance in neonatal care and healthy intellect during later developmental period. Acknowledgment: This work is supported by grants from DST, DBT, ICMR and KSCSTE

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Abstract No: 1.12

Diagnosis of Organic Acidurias in Children - 2 Years Experience in a Tertiary Care Center

M P Narayanan, Kannan Vaidyanathan and D M Vasudevan Metabolic Disorders Laboratory, Dept. of Biochemistry, Amrita School of Medicine, Amrita Vishwa Vidyapeetham, Cochin-682041, Kerala, India. Email: [email protected]

Objectives 1) . To screen 420 patients with signs and symptoms of organic aciduria / aminoaciduria. 2) To identify and quantify organic acids and amino acids by HPLC. Methods: Manual screening tests and thin layer chromatography were used to detect the presence of abnormal metabolites in urine. HPLC was done for organic acid and amino acid analysis using standard protocols. Clinical details of the patients were collected from proforma. CT/MRI were used to evaluate the brain abnormalities. Results: We studied 420 children upper age limit 12 years with signs and symptoms of organic acid / amino acid disorders. Abnormal cerebral CT/ MRI findings were found in 11 patients, and abnormal EEG findings in 63 patients. 110 samples found positive for organic acids by initial Abstract No: 1.13

screening tests. Out of which 41(9.7%) cases of Organic acidurias were diagnosed by HPLC. 15 cases of Methylmalonic aciduria, 16 cases of Propionic aciduria, 7 cases of Maple syrup urine disease, 2 cases of Alkaptonuria and 1 case of Isovaleric aciduria were found. Common symptoms of Organic aciduria patients were seizures (N=17), delayed milestones (N=12), skin rashes (N=10), abnormal muscle tone (N=6), poor feeding (N=12), mental retardation (N=12), encephalopathy (N=1) and hyperammonemia (N=30). 10 patients were from consanguineous families. 15 patients expired during the study. Conclusion: Methylmalonic aciduria, Propionic aciduria and Maple syrup urine disease account for 90% of organic acidopathies detected. HPLC quantification of amino acids and organic acids helps to identify organic acidurias. Prompt diagnosis allowed specific treatment in majority of these patients with rapid improvement.

Detection of Inborn Errors of Metabolism in Paediatric Age Group in a Tertiary Care Centre.

Saritha KamathU1, Licy C.D1, Nalini Bhaskaranand2 and Anjali Rao3 1 Department of Medical Laboratory Technology, MCOAHS, 2 Department of Paediatrics, 3Department of Biochemistry, KMC, Manipal. Objective: Inborn errors of metabolism (IEM) may occur anywhere in the biochemical machinery of living cells. Although some 150-200 inborn errors of metabolism of clinical interest are known at present, a few thousand enzymes and structural proteins have so far been described

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in human cells; thus it is likely that the types of inborn errors so far described represent only a fraction of those actually occurring. A large number of inborn errors of metabolism in children remain undetected in India due to lack of awareness, investigative facilities and economic constraints.

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Methods: We examined 21 children presenting protocol. We have come across following cases in with neurological and metabolic problems at a the course of our study- methyl malonic aciduria, tertiary level teaching hospital in Karnataka for homocystinuria, and maple syrup urine disorder. inborn errors of metabolism using a standard Abstract No: 1.14

Hyperprolinemia in 22q.11 Deletion Syndromes and its Association with Neurological Deficits

Sweetha Mary Mammen1, Dr. K.N. Subhakumari1 and M. V. Thampi2 1 Department of Biochemistry, 2Department of Human Cytogenetics, Amrita Institute of Medical Sciences, KERALA Type 1 hyperprolinemia is biochemically characterized by a defect of the proline dehydrogenase enzyme (McKusick 239500). The enzyme has been mapped to the PRODH gene on the human chromosome 22 on band q11.2. This gene falls in the region deleted in the 22q11 Deletion Syndrome, which includes DiGeorge and Velo-cardio-facial syndromes. Neurological manifestations have been reported in several hyperprolinemic subjects. The paucity of previous research in this subject in India is investigated this study. Objectives: Methods: Plasma proline levels were assayed (using high pressure liquid chromatography) in 14 children affected with 22q11 deletion syndrome. The neurological and development statuses of the hyperprolinemic subjects were studied. Results: Seven subjects had proline levels above normal and seven were within the normal limits. Three subjects had varying degrees of psychomotor deficits, including delayed milestones, global developmental delay and decreased IQ.

Summary: In concordance with previous studies, this study finds that hyperprolinemia is a common finding in 22q11 deletion syndrome. Besides, it is 1 To study the frequency of type 1 in agreement with prior studies, stating an hyperprolinemia in patients diagnosed with 22q11deletion syndrome at Amrita Institute of association between hyperprolinemia and neurological disorders. A low proline diet is Medical Sciences, Kerala, India. suggested since prior research has shown that it 2. To examine the association between plasma improves growth and development in proline levels and clinical features (neurological hyperprolinemic individuals. deficits) of the above patients. Abstract No: 1.15

Hypoglycaemia

Biswajit Saha Durgapur Steel Plant Hospital, Durgapur ­ 713 205, West Bengal (India); e-mail : [email protected] Hypoglycaemia is a medical emergency. In day caused by "over administration" of antidiabetic to day clinical practice, this condition is commonly drugs. Hypoglycaemia is defined when the plasma

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glucose level is usually less than 45 ­50 mg/dl. However, chemical hypoglycaemia is not synonymous with clinical hypoglycaemia which requires demonstration of Whipple's triad ­ symptoms consistent with hypoglycaemia, documentation of a low plasma glucose concentration and relief of symptoms after plasma glucose level has been raised. Interestingly, the threshold for hypoglycaemia varies widely in an individual diabetic patient as a result of alteration Abstract No: 1.16

of metabolic setpoint from time to time in different clinical settings. Relative hypoglycaemia occurs with blood sugar level above 50mg/dl. Conversely, remarkable lowering of threshold is found in diabetic patients as a result of recurrent attacks of hypoglycaemia. Surprisingly, in some otherwise normal (nondiabetic) individuals, marked lowering of blood glucose level is not associated with any symptom(s). The details of causes, clinical manifestations and management will be discussed.

Dyslipidemias

Prof. N. Haridas Professor and Head, Department of Biochemistry PS Medical College, Karamsad The Plasma lipoproteins are spherical macromolecular complexes of lipids and specific proteins (apoproteins or apolipoproteins). The lipoprotein particles include chylomicrons, very low density lipoproteins (VLDL), low density lipoproteins (LDL) and high density lipoproteins (HDL).They differ in lipid composition, size, density and site of origin. Lipoproteins function both to keep their component lipids soluble as they transport them in the plasma and to provide an efficient mechanism to transport their lipid components to (and from) the tissues. In humans, the transport system is less perfect than in other animals and, as a result, humans experience a gradual deposition of lipid-especially cholesterolin tissues. This is a life threatening occurrence Abstract No: 1.17 when the lipid deposition contributes to plaque formation, causing the narrowing of blood vessels (atherosclerosis). The talk shall include a discussion on the metabolism of lipoproteins and lipids and how their metabolism alters in diseases like diabetes mellitus, metabolic syndrome, familial hyperlipo-proteinemias, obesity and athero-sclerosis. Mention also shall be made about blood investigations like lipid profile and other risk factors like lipoprotein `a', homocysteine, creactive protein and dietary fat. Certain preventive measures like dietary modifications to ward off atherosclerosis and subsequent heart ailments would also form a part of the discussion.Certain interesting observations on lipd profile soon after acute myocardial infarction shall be discussed.

Effect of Swimming on Selected Health Parameters of Beginners

Ragini Srivastava, Royana Singh, Ratna Pandey and Amit Srivastava Department of Biochemistry, Institute of Medical Sciences, Banaras Hindu University, Varanasi. Email- [email protected] Background: Exercise has been shown to be a simple and economical preventive modality that may be considered as a beneficial adjuvant for

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many life style diseases(LSD). This study has been done to investigate the impact of swimming on biochemical, anthropometric and physiological

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parameters representing health and fitness in a high-density lipoprotein (HDL) were determined. short period. Anthropometric parameters related to girth and width measurements like height, weight, BMI, Methods: This prospective randomized study body fat mass and chest circumference were taken. consisted of 30 cases in the swimmers group that Physiological parameters related to pulmonary was matched with 26 controls in non-swimmers group. Biochemical parameters such as fasting function test were measured at baseline and after blood glucose (FBG), serum total cholesterol 10 weeks intervals. Results: Study yet to be (TC), triglycerides, low-density lipoprotein complete and therefore results will be discussed (LDL), very low-density lipoproteins (VLDL) and at the time of presentation.

Abstract No: 1.18

Evidence-Based Guidelines for the use of Biochemical Markers of Bone Turnover in the selection and monitoring of Bisphosphonate Treatment in Osteoporosis: A Consensus Document of the Belgian Bone Club

Dr.Maulik Nayak, Dr. Alok Parekh, Dr.Rita Shah and Dr. D.A. Trivedi

Objective: To review the clinical value of bone turnover markers (BTM), to initiate and/or supervise anti-resorptive management for osteoporosis compared with bone mineral density(BMD) and to evaluate proper BTM and changes in BTM levels for significance of treatment efficiency. Methods: Consensus meeting generating guidelines for clinical practice after review and discussion of the randomized controlled trials or meta-analyses on the management of osteoporosis in postmenopausal women. Results: BTM cannot be used as predictive markers of BMD in an individual patient even if the correlation between BMD & BTM is statistically significant. Both are independent predictors of fracture risk, but BTM can only be used as additional risk factors in the decision to treat. The use of BTM select for optimal treatment

not supported with current records. However, they can be used to monitor treatment efficiency before BMD changes can be evaluated. Early changes in BTM can be used to measure the clinical efficacy of an anti-resorptive treatment and to reinforce patient compliance. Discussion: An optimal long-Term effect is not evident with determining BTM reflecting threshold. The objective should be return to the premenopausal range and / or a decrease at least equal to the least significant change (30%). Preanalytical & analytical variability of BTM is important limitation to their use. Conclusion: Consensus regarding the use of BTM resulted in guidelines for clinical practice. BMD determines the indication to treat osteoporosis & reveal treatment efficiency. It can motivate patients to persist their medication.

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Abstract No: 1.19

Requirement of Essential Fatty Acids: An Analytical Approach

Dr. Sankar Roy Asst. Prof., Tripura Medical college & Dr. BRAM Teaching Hospital, Hapania , Agartala- 799014, Tripura Each animal or plant species has a specific pattern of fatty acid composition in their body. Human body contains more unsaturated FA(60%) than saturated FA(30%). This ratio (2:1) is essential to maintain structural and metabolic function of cell membrane. Body synthesizes Eicosa pentaenoic acid, Docosa hexaenoic acid and Prostaglandins from essential fatty acids. But if the dietary PUFAs are in excess , they compete with or inhibit phospholipases and acyltransferases. The dietary Abstract No: 1.20 PUFAs are incorporated into the structural lipids about 20 times more than the ve novo synthesized long chain fatty acids. Ratio of w-6 / w-3 is also to be maintained about 1:1. But modern diet contain more than 10:1. Therefore, it is recommended to consume more w-3 fatty acid containing foods like Soybean oil (7%), vegetable oil, marine fish to equalize w-6 but not in excess amount.

Clinical Approach to Integrated Basic Sciences Teaching

Shruti Mohanty Department of Biochemistry, Kamineni Institute of Medical Sciences, Narketpally. Email: [email protected] teaching methodology. Evaluation of the Clinical case solving valuated the higher order cognitive skills of the learners.

Objectives: 1.To compare the performance of students experiencing integrated teaching with those exposed to traditional didactic lectures without integration. 2. To evaluate the acceptability of integrated teaching and clinical case discussion to first year medical students.

Results: The pre and post test scores of both the group experiencing integrated teaching and traditional didactic lectures were compared. The Method: Nine faculty members trained earlier mean pre and post test scores in the group that during in house ME workshops volunteered to underwent intervention were 4.5/15 and 9/15 participate in the project. Three modules were respectively whereas the pre and post test scores designed and implemented on fifty alphabetically of the group exposed to traditional teaching were selected students of the 150 enrolled in first year 4.1/15 an 6/15 respectively. Moreover forty two MBBS course. Pre & Post test scores were of fifty students who experienced the intervention analyzed to determine the relative impact of the solved the cases correctly but only twenty two two teaching approaches on student learning. Pre students out of fifty could solve correctly in the validated questionnaires for students and faculty group that experienced traditional teaching. were introduced to assess their perceptions on the Majority (88%) of the students experiencing

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integrated teaching agreed that the integrated approach helped them correlate across Anatomy , Physiology and Biochemistry. 96% of students and 100% (9) faculty expressed that integrated teaching helped the students understand and therefore retain the subject better. Likewise, 96% students and 100% faculty expressed that this Abstract No: 1.21

method of teaching will certainly help the students perform better in examination. Conclusion: With minimal intervention of juxtaposing similar topics and having small group discussions the students' performance in examination improved two fold when compared to the traditional method.

A Study on Organic Aciduria: Detection and Quantification using HPLC

Aswathi K.I, Narayanan MP and Kannan Vaidyanathan Dept of Biochemistry, Amrita School of Medicine, Kochi, Kerala, INDIA. Email: [email protected] Objective: Organic aciduria is a group of disorders characterized as autosomal recessive. It is primarily due to defect of an enzyme, which results in block in a particular metabolic pathway. A pilot study was done to identify and quantify the different Organic acids using HPLC. Methods: The study was carried out at Amrita Institute of Medical science and Research centre, Kochi. 184 urine samples were taken from Organic aciduria suspected children between the ages of 0 ­ 15 years during a time period of one year from May 2008 ­ April 2009. Initially samples were screened for Organic acids by thin layer chromatography. This was followed by HPLC to confirm the presence and to identify and quantify the individual Organic acids. Abstract No: 1.22 Results: For the study, 184 urine samples were screened, 39 showed organic aciduria by screening. 26 were confirmed as positive cases on further detection using HPLC. There were 34.6% cases of Propionic aciduria, 26.9 % cases of Methylmalonic aciduria and 7.6% cases of lactic aciduria. Conclusion: This study shows that the incidence of Organic aciduria is high and HPLC helps in the easy detection of individual organic acids present in the urine from which the metabolic disorder can be confirmed. Thereby, early treatment can be initiated to avoid morbidity and mortality.

ELISA based Evaluation of 17-á Hydroxy Progesterone as a Diagnostic Marker for Congenital Adrenal Hyperplasia and Assessment of Hyperhomocysteinemia in Coronary Artery Diseases and Cerebrovascular Accidents

Jayakumar N., Dr. Sujaya P and Dr. Kannan Vaidyanathan Department of Biochemistry, School of Medicine, Amrita Institute of Medical Sciences, Kerala. [email protected] Background: Congenital Adrenal Hyperplasia synthesize cortisol, and in most instances, also the (CAH) is a family of autosomal recessive inability to synthesize aldosterone. disorders characterized by the inability to Hyperhomocysteinemia, has recently been

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associated with increased risk of vascular disease. The effect of elevated homocysteine is multifactorial, affecting both the vascular wall structure and the blood coagulation system. Objective: To assay 17 ­ OHP levels in patients suspected with Congenital Adrenal Hyperplasia using ELISA & to evaluate its use as a Diagnostic marker for Congenital Adrenal Hyperplasia. Assessment of Hyperhomocysteinemia as a risk factor for Coronary Artery Disease & Cerebrovascular Accident.

Results: 53/160 subjects showed elevated 17hydroxy progesterone (33%) levels. Based on the clinical findings, out of 160 patients, 73 patients were diagnosed as having Congenital Adrenal Hyperplasia. (45%). 51/170 patients showed elevated homocysteine (30%) levels. Among the observed 51 hyperhomocysteinemic subjects, 21 of them were affected by stroke (41%).

Conclusion: This study suggests that Hyperhomocysteinemia is associated with Cerebrovascular Accidents and Coronary Artery Disease and that it is an independent risk factor Methods: The DEMEDITEC 17- á -OH for the same. Besides, the basal measurement of Progesterone Enzyme Immunoassay kit was used 17-OHP can be considered as a valuable test for for the assay of 17-á-OH progesterone in serum the initial diagnosis of CAH, however one has to of160 patients suspected of CAH. Bio-Rad Micro correlate with the clinical symptoms for the plate Enzyme Immunoassay Homocysteine Test confirmation. was used for the estimation of homocysteine in 170 patients. Abstract No: 1.23

Standardisation of Chromatographic Techniques: HPLC & GC ­ A Diagnostic Tool for Inborn Errors of Metabolism

Ms. Amalmol Peter and Dr. Kannan Vaidyanathan Department of Biochemistry, AIMS, Cochin. Inborn Errors of Metabolism are hereditary affections resulting from incompetence in enzymatic reactions of intermediary metabolism. At present, several hundred hereditary metabolic disturbances are known, many of which correspond to severe life-threatening disorders. Many IEM have similar clinical presentations, making the early diagnosis difficult. Procedures for the qualitative detection and quantitative determination of diagnostic markers are based on a variety of methods.Chromatography has played a pivotal role in the diagnosis of IEM. Objective: This study aims to standardize HPLC& GC to the analysis of organic acids & amino acids for the detection of Inborn Errors of Metabolism.

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Methods: The study was carried out at Metabolic Disorders Laboratory, Department of Biochemistry, AIMS, Cochin, during the period 10.12.2008 to 30.04.2009.For standardizing HPLC, 17 aminoacids & 6 organic acids were tested at varying concentrations and the analysis was performed on Shimadzu HPLC ­ DAD LC 10.For standardizing GC, 5 organic acids were tested using varying concentrations & the analysis was performed on Shimadzu GC- FID 2014. Results: Validation studies for aminoacids & organicacids by HPLC showed better sensitivity and steability at retention times than other methods. Both the standard amino acid and the organic acid showed a good linearity for 3 different

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concentrations.Validation studies for organic acids by GC using BSTFA: TMCS (Bis - trimethylsilyl trifluoroacetamide: Trimethyl chlorosilane) derivatizing agent showed faster separation than other methods. Conclusion: From the present study, it can be concluded that, the methods described in this study Abstract No: 1.24

permitting both the identification & quantification of organic acids & amino acids in urine, plasma or amniotic fluid for the diagnosis of Inborn Errors of Metabolism with good precision & sensitivity, minimal manual handling and the advantage of a more versatile apparatus.

Faecal Calprotectin in Inflammatory Bowel Diseases

M.P. Narayanan*, Kannan Vaidyanathan*, O. Gajendra#, S. Shine#..and V. Balakrishnan# *Dept. of Biochemistry, Amrita School of Medicine, Kochi-682041, Kerala India, #Dept. of Gastroenterology, Amrita Institute of Medical Sciences and Research Centre, Kochi-682041, Kerala, India. [email protected] Background: Inflammatory bowel diseases (IBD) are chronic intestinal disorders of unknown etiology and with a typically relapsing course. Faecal calprotectin, an important granulocyte cytosolic protein, is closely correlated with faecal excretion of 111 indium labelled leucocytes, deemed to be the gold standard for measuring intestinal inflammation. Assessment of faecal calprotectin levels has been proposed as a noninvasive test for the direct evaluation of intestinal inflammation in patients with IBD. Objective: To compare faecal Calprotectin with the standard disease activity indices (UCAI and CDAI) of inflammatory bowel diseases (Ulcerative colitis and Crohn's disease). Methods: Patients diagnosed to have IBD based on clinical, endoscopic and histological examination were included. Ulcerative colitis activity index (UCAI) and Crohn's diseases activity index (CDAI) were calculated. Faecal calprotectin was estimated by a commercially available quantitative ELISA test. Results: Forty-three patients were included in the study, 20 patients with Ulcerative colitis (UC) and 23 with Crohn's disease (CD). Patients with active CD (CDAI > 150) were 18/23 (78%) and with active UC (UCAI > 2) were 17. Mean hemoglobin was not different in both the groups. Mean ESR was raised in both groups (37 in UC, 31 in CD; P = 0.361). Mean CRP was raised in both groups (UC 49± 60; CD 19±19; P= 0.302). Mean UCAI was 7 (SD± 3) and mean CDAI was 212 (SD ± 89). Mean faecal calprotectin was 890 (SD±503) in UC patients and 641 (SD±739) in CD patients (P = 0.028) Fecal calprotectin was higher in active cases compared to those in remission but the difference did not achieve statistical significance. Correlation of faecal calprotectin with CDAI was strong (P = 0.0008) whereas correlation of faecal calprotectin with UCAI was weak (P = 0.274). Conclusion: Faecal calprotectin correlated strongly with CDAI but weakly with UCAI. The difference in patients in remission vs active disease (as categorized by UCAI and CDAI) was not statistically significant.

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Abstract No: 1.25

A Study on Cytotoxicity and Inhibition of Cyclooxygenase-2 by Curcumin and Diallyl Sulfide in Head And Neck Cancer Cell Line

Hariharasudan. R Final year MBBS, Amrita Institute of Medical Sciences, Kochi , Kerala. Email: [email protected]

Objective: To study the effect of curcumin and diallyl sulfide (DATS) on cytotoxicity and inhibition of Cyclooxygenase (COX-2) in head and neck cancer cell lines Methods: MDA 686LN a HNSCC cancer cell line, maintained in MEM was teated with DATS and curcumin individually for 24,48 and 72 hours with a zero hour control in triplicates. The cells were harvested by trypsin treatment and cell viability was determined by trypan blue dye exclusion assay. Total RNA was isolated using TRIZOL solution and its integrity was checked. The cDNA was synthesized using both oligo-dT and random hexamer primers. The level of COX1 and COX-2 expression was analyzed by PCR in comparison with â-actin using specific primers. Abstract No: 1.26

Results: The study showed that the IC50 values for both DATS and curcumin was 5ì M. With regard to the regulation of COX-1 and COX-2 expression, curcumin and DATS inhibited COX-2. Conclusion: Currently, curcumin has been introduced into clinical trials in the chemoprevention of HNSCC and the only difficulty is its bioavailability. The cytotoxicity study, using the HNSCC cell line namely MDA 686LN indicated that curcumin and DATS have greater effect on cell viability. It is proposed that, the synergistic effect of the combination of curcumin and DATS should be greater than the individual clinical compounds. Further studies in this aspect are currently taken up. Acknowledgment: : This work was done as a part of ICMR ­ STS project.

A Study for Investigating the Association between Semen Biochemical Parameters and Routine Clinical Semen Parameters in Infertile Men

*

Suresh S. and T. Vijayakumar Department of Biochemistry and Physiology , Educare Institute of Dental Sciences, Malappuram-675 506, Kerala ,India. Objective: The biochemistry of semen has been studied extensively,but the biochemical analysis of semen has little part to play in the routine investigation of an infertile man. 1-4 In this context a study was undertaken with the objectives of investigating whether there exists an association between semen biochemical parameters and clinical semen parameters, so that the latter could

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be used as an adjunct to the conventional semen analysis for evolving better seminogram protocols in the evaluation of male infertility. Methods: For the present study , semen samples of 120 infertile men visiting an Infertility Clinic were collected and the same was used for the investigations. The study followed the ICMR and

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WHO Ethical Guidelines for Clinical Research.The 120 infertile men were divided in to 3 groups: Normospermic, Asthenospermic and Oligospermic subjects, based on the sperm count and sperm motility. There were 32 normospermic, 51 asthenospermic and 37 oligospermic subjects. The grouping was based on the WHO guidelines for semen analysis. The results of clinical parameters such as sperm motility, viable count and pH were collected from the clinical proforma. The semen biochemical parameters investigated in the present study were prolactin, acid phosphatase, sorbitol dehydrogenase, glucose, fructose and pyruvate.

Conclusion: Thus, this study reveals that there exists a strong relationship between the biochemical parameters studied and the routine semen clinical parameters of asthenospermic male subjects with clinical infertility . Although the present study could not find such a relationship in oligospermic subjects with clinical infertility, additional semen biochemical components such as citrate, choline and spermine as well as the semen anti-oxidant system should also be analyzed before rejecting the hypothesis that a similar association does not exist in oligospermic males with clinical infertility.

1. Insler.V, Lunenfeld.B.: Infertility: the dimension of the problem, in Insler.V, Lunenfeld.B (eds) Results: The results were analyzed statistically Infertility male and female(2) Peter Richardson using Mann-Whitney U statistics in the Sigma 1993, Pg.3-5. Stat 3.5 Version Software. A statistically significant decrease in all semen biochemical parameters such as prolactin, acid phosphatase, 2. Attramadal. A, Tveter. K.J, Weddington. S.C. et al.: Androgen binding and metabolism in human glucose,fructose, pyruvate and sorbitol prostate. Vitam Horm 1975, (33) pg.247. dehydrogenase were observed in asthenospermic subjects as compared to normospermic subjects ( 3. Pulkkinen. P, Sineroiste. R, Janne. J.: p <0.001 ) A statistically significant decrease in Mechanisms of action of oxidized polyamines prolactin & acid phosphatase and increase in on the metabolism of human spermatozo J glucose were observed in oligospermic subjects Reprod Fertil1978, (51) pg.399. as compared to normospermic subjects ( p <0.001).A statistically significant increase in 4. Sheth. A.R, Mugtawala. P.P, Shah. G.V, Rao. SS.: prolactin and decrease in glucose, fructose and Occurance of prolatctin in human semen. Fertile pyruvate were observed in oligospermic subjects steril 1975, (26) Pg. 905. as compared to asthenospermic subjects ( p <0.001). Abstract No: 1.27

Isolation and Validation of the Biomarker MMP1 from Salivary Transcriptome for Early Detection of Oral Cancer

Kannappan Alagappan, Amritha Suresh and M A Kuriakose Amrita Institute of Medical Sciences, Kochi. Background: Oral cancer incidence is on the rise worldwide with a marginal improvement in survival rates over the past few decades which is due to their late presentation at the time of diagnosis. Early detection would be the most effective means to reduce death from this disease and in this regard expression profiling of biomarkers in saliva appears to holds promise. Methods: Saliva was collected (1ml) in RNA later from patients and normal controls and RNA isolated using QIAamp viral RNA mini kit. DNA

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contamination was checked by PCR and if contaminated, DNase treatment was done. Conversion of the RNA into cDNA was done and the integrity of cDNA was confirmed by RT-PCR using GAPDH primers. The expression of MMP1 in saliva was analyzed using specific primers by semi-quantitative RT-PCR.

patients, one of them progressed to oral cancer on follow up.

Conclusion: The main aim of a cancer-screening program is to detect the tumor at an early stage in order to achieve a better prognosis. The medium used, if non-invasive and inexpensive like saliva; the compliance and frequency of the screening Results: Saliva was collected from 15 patients process can be increased. MMP1 over expression divided into three groups with 5 subjects in each; in saliva found in this study is suggested to normal controls, patients with pre-malignant delineate the lesions with a high risk of lesions and oral cancer. Out of these 15 samples malignancy. 10 showed MMP1 amplification; 4 normals, 2 preAcknowledgment: This wark was done as part malignant and 4 oral cancers. Patients showed of ICMR-STS Fellowship MMP1over expression as compared to the basal expression in normal controls. Of the 2 PML

Abstract No: 2.1

Serum Copper and Magnesium Level in Type ­2 Diabetes Mellitus

B.K.Agrawal1, Seema Tamrakar1 and V.K.Sharma2 1 Dept. of Medical Biochemistry, 2Dept. of Medicine. Gandhi Medical College , Bhopal,E-mail:[email protected]

Objective: Diabetes Mellitus is a chronic metabolic disorder, copper level is increased and magnesium decreased in Diabetes. Methods: The present study includes 100 patients of Diabetes Mellitus compared to 50 healthy persons. Patients were selected randomly without any bias for age sex. 1.Serum copper estimated by colorimetric method. 2.Serum magnesium estimated by calmagite method. Results: The mean values of copper was found to be 110.04±13.28µg/dl and magnesium was 1.84±0.45MEq/L in healthy subjects type-II DM

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The mean value of Copper was found to be 220.04±28.46µg/dl and magnesium was 0.6±0.49 MEq/L in controlled Diabetic patients. The mean value of copper was found to be 225.87±23.45 /dl and magnesium was 0.77±0.69MEq/L in uncontrolled Diabetes. Serum copper level was significantly increased compared to control group (p<0.001), whereas serum magnesium level was significantly decreased (p<0.05) compared to control group. Conclusion: It can be concluded that diabetes is also associated with hypercupremia and hypomagnesemia.

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Abstract No: 2.2

Biochemical Profile of Cord Blood in Normal Pregnancy and Pregnancy Induced Hypertension

Miss D.Prathima , Dr. U.B.Vijaya lakshmi and Prof. Dr. K. Ambika Devi Dept. of Biochemistry, ASRAM Medical college , Eluru, A.P; Email : [email protected] Background: Pregnancy induced hypertension (PIH) is a significany health problem and is the leading cause of maternal and perinatal mortality and morbidity. Low birth weight and prematurity is common in such cases. Objective: This study was designed to compare certain biochemical parameters in cord blood of children born to mothers with PIH and to mothers with normal pregnancy. Methods: For this purpose 10ml of cord blood during delivery was collected from mothers attending obstetrics department at ASRAM medical college, Eluru.30 samples were collected from mothers with uncomplicated pregnancy and 50 samples from the mothers with PIH (maternal age matched). The 50 samples include group A20 mothers with pre-eclampsia and group B-30 mothers with gestational hypertension The Abstract No.2.3 biochemical parameters measured were glucose, urea, total proteins, total cholesterol, calcium & magnesium. Results:It was observed that all the parameters except for total cholesterol and calcium were significantly lower (p<0.00001) in the cord blood of babies born to mothers with PIH.The total cholesterol and calcium were significantly higher in the same cases (p<0.001). On comparison of parameters of cord blood in babies weighing > 2.5kg in cases vs controls, there was significant decrease in all the parameters in the former. Conclusion: So the neonates born to mothers with PIH must be screened carefully to prevent mortality and morbidity due to low levels of the important biochemical parameters like glucose and magnesium. Neonates weighing >2.5kg are also at similar risk.

Relationship between Micronutrient Intake of Mothers and Metabolic Outcomes in Offspring (6 and 12 yrs): Pune Maternal Nutrition Study

S.S.Naik, D.S.Bhat, C. Joglekar and C.S.Yajnik K.E.M.Hospital Res. Centre, Pune 411000 Objective: Role of micronutrients during the maternal food intake, circulating nutrients and pregnancy on the development of adiposity in the physical activity twice during pregnancy. children at 6 and 12 yrs. Anthropometric measurements on 690 children born to these mothers were carried out at birth and Methods: PMNS is a prospective study of then every six months till 6 yrs and then yearly till maternal nutrition and fetal growth. We recorded 12 yrs. At 6 and 12 yrs fasting plasma insulin, leptin

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and lipids were measured. Ethical approval and consent was obtained. Results: There was no gender difference in weight or BMI at 6 and 12 yrs. Girls had higher leptin, and insulin concentrations at 6 yrs (3.1 vs 2.4 and 3.3 vs 3.03 mU/L respectively) and at 12 yrs (3.9 Vs 2.4 and 5.7 vs 4.7 mU/L respectively). BMI, insulin and leptin concentrations were directly associated with adiposity at 6 and 12 yrs (p<0.001 for all). Those who were born thin and grew rapidly from 3 yrs had higher plasma leptin and insulin concentrations (p=0.001). Higher maternal weight gain during pregnancy predicted higher leptin concentration (p=0.04). At 12 yrs, leptin Abstract No: 2.4

concentration was directly associated with weight and inversely with height even after adjusting for maternal micronutrients, adiposity at birth and adiposity at 12 yrs (p=0.017 and -0.038 resply). Higher maternal BMI and erythrocyte folate concentration predicted higher adiposity and leptin concentration at 12 yrs (p=0.04 for both). Conclusion: Maternal micronutrient intake and high erythrocyte folate concentration in pregnancy predict higher adiposity, insulin resistance, and leptin concentration in the offspring. The effect is stronger at 12 years, suggesting intrauterine nutritional programming of adiposity

Lack of Awareness Resulting in Lead Poisoning in Unorganized Sector of Battery Workers

Dr. Vishal Babu G N, Mr. Raviraja A and Dr. T Venkatesh Department of Biochemistry, St. John's Medical College, Bangalore, India

Objective: A comparative study of renal functions and blood pressure of organized and unorganized sector of battery workers was conducted to study the effect of lead exposure and lack of awareness between the organized and unorganized sector. Methods: Ninety subjects were selected for the study of which: Group A had 30 Controls; Group B had 30 organized battery workers and Group C had 30 unorganized battery workers. They were evaluated for their Blood lead level, Zinc Protoporphyrin and renal functions along with measurement of blood pressure. Results: This study shows that there is statistical significance between Group B and Group C in

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Blood Lead level (p<0.001), Zinc Protoporphyrin (p<0.001), Urea (p<0.001) and Uric Acid (p<0.001) Conclusions: There was significant difference in blood lead level, Zinc protoporphyrin and renal parameters between the two sector though there was no significant difference between the two in terms of blood pressure. This difference was mainly due to the absence of precautionary principles, proper disposal methods and proper education to workers in the unorganized sector. This study signifies the importance of all these principles.

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Abstract No: 2.5

Estimation of Zn, Mg in Several Protein Energy Malnutrition in Children Age Group 6 Month to 5 Year

Farha Ali1, B. K. Agrawal1 and Manjusa Goel2 Department of Biochemistry, 2Department of Paediatrics, Gandhi Medical College, Bhopal, Email: [email protected] Results: The mean value of serum Zinc (in age group 6month to 2yr and 2 yr to 5yr) were found to be significantly (P<0.001) decreased as compared to control) were found to be significantly.(P<0.001) decreased as compared to Methods: 50 PEM children were studied control. Results obtained from analysis of Mg and compared to 20 healthy controls divided on the Zinc level were significantly low. basis of age group 6month to 5yr. Zinc was Conclusion: Thus it can be concluded that, there estimated calmagite method. Mg was estimated is sever deficiency of Zinc and Mg in PEM children by colorimetric method. due to their abnormal metabolism. Objective: Malnutrition is the cellular imbalance between the supply of nutrition and energy and the body demand for them to insure growth maintenance an specific functions. Abstract No: 2.6

Zn and Fe Levels in Hypothyroid Subjects: A Hospital Based Case Control Study

Gaurav B. Shetty1, Suchetha Kumari. N 2 and Smitha.P 3, 1 IIIrd M.B.B.S K.S Hegde Medical Academy ,Deralakatte , Mangalore ­575018, 2Professor, Dept.of Biochemistry, K.S Hegde Medical Academy, Deralakatte, Mangalore ­575018, 3M.Sc Biotechnology SDM College Ujire. Email: [email protected] Background: Excess or decreased secretion of thyroid hormone leads to hyperthyroidism and hypothyroidism respectively. Iron is a very critical mineral for individuals with thyroid disease. Zinc is one of the most abundant trace elements in the body. Zinc has close interrelationship with the endocrine system, it is essential for normal growth, reproductive function, thyroid function and glucose metabolism. Methods: This study was carried out at K.S.Hedge Medical College Hospital. Individuals were diagnosed as hypothyroid and normal by ELISA method .A total of 40 subjects were included in the studies among them 20 subjects with hypothyroidism and 20 subjects with normal thyroid function. Iron was estimated by Bathophenanthroline method and Zinc by Atomic absorption spectrophotometer. Results: Iron and Zinc levels were significantly decreased in hypothyroidism subjects when compared to normal subjects. Conclusion: Our study shows decreased levels of iron and zinc in hypothyroidism, this may be due to deficiency in thyroperoxidase activity and copper level which could interfere with thyroid hormone synthesis. Hence Iron and zinc supplements may improve thyroid metabolism in individuals with hypothyroidism.

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Abstract No: 2.7

Extracellular Mg2+ Decreases Loading of the Ca2+-Sensitive Fluorescent Probe Chlortetracycline, into Calcium Stores in Lymphocytes and Neutrophils

Nivedita L. Rao1 and Jose Jacob2 1 Department of Biochemistry, Yenepoya Medical College, Yenepoya University, Mangalore-575 018, Karnataka, 2Department of Biochemistry, Amala Institute of Medical Sciences, Thrissur-680 555, Kerala; E-mail: [email protected] Objective: Extracellular Mg2+ is known to inhibit agonist-stimulated calcium mobilization from intracellular calcium stores and exocytosis in several cell types, possibly by decreasing the free Ca2+ content of the stores. Methods: We used Chlortetracycline (CTC), a Ca2+-sensitive fluorescent probe for cellular calcium stores, to study the effect of extracellular magnesium on the quantity of CTC loaded into cells which in turn, depends on the free Ca2+ content of the stores. CTC was loaded into freshly isolated, viable, mouse spleen lymphocytes and human blood neutrophils. Estimation of loaded CTC was done by digitonin-permeabilization of CTC-loaded cells, to release the loaded CTC and comparing the Ca 2+ -induced fluorescence enhancement of CTC with a standard plot. In the presence of 1 mM extracellular Ca2+, significant CTC-loading occurred. Abstract No: 2.8 Results: Absence of extracellular Ca2+ (and Mg2+) during CTC-loading caused a decrease in the quantity of CTC loaded into cells. Presence of 1 mM extracellular Mg 2+ (in the absence of extracellular Ca2+) caused a further decrease in CTC-loading. Extracellular Ca2+ replenishes Ca2+ at the stores. Therefore, in the absence of extracellular Ca2+ (and Mg2+) the quantity of free Ca2+ at the stores, decreased and consequently, CTC-loading into those cells decreased. Further decrease in CTC-loading in the presence of extracellular Mg2+ (and absence of extracellular Ca2+) indicates decreased relative free Ca2+ content at cellular calcium stores. Conclusion: These results have general importance in relation to the modulatory role of Mg2+ on intracellular Ca2+ signaling processes and the high Mg2+-induced suppression of several biological functions of cells

Expression of Metallothionein (MT) in Colonic Precancerous and Cancerous Tissue in Animal Model

Pamela Christudoss, and R. Selvakumar Dept. of Clinical Biochemistry, Christian Medical College ­Vellore. Email id: [email protected] Objectives: 1) To study the MT protein content, protein expression and mRNA expression in association with alterations in tissue zinc levels in DMH (dimethyl hydrazine) induced colonic precancerous and cancerous tissue in animal model. 2) To study the effect of Aspirin, vitamin

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C or zinc supplementation on the metalothionein mRNA expression, protein expression and content in colon carcinoma in rat. Methods: Rats were assigned to 2 groups: control and test group which received saline and

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dimethylhydrazine respectively for 4 months and sacrificed at six months. The colon of the rats of both groups were harvested and fixed in 10% buffered formaldehyde and processed for histology study and for immunohistochemistry of metallothionein expression as per standard methods. A part of the colon was used for MT mRNA expression by mRNA Isolation followed by cDNA conversion and quantitative real time PCR. Tissue zinc was estimated by Atomic Absorbtion Spectrophotometry.

mean MT mRNA expression was actually increased to a level much higher i.e. up to 1.9 fold and up to 2.4 fold respectively than the saline treated control rats. When the DMH treated rats were co-treated with zinc, a mean increase in MT mRNA expression up to 2.7 fold and 3.25 fold was observed in the precancerous group and cancerous group resp. as compared to the saline treated control rats. When the DMH treated rats were cotreated with aspirin, vitamin C or zinc, the MT showed a mean immunopositive staining of 58%, 60% and 75% resp; but this did not reach the Results: Colonic tissue zinc in DMH treated rats normal MT immunopositivity levels. decreased by 60-70% as compared to saline treated controls. The mRNA expression in the DMH Conclusions: This study shows that zinc decrease induced precancerous and carcinoma model was due to the onset of carcinogenesis co-related with significantly decreased by 1.33 fold (mean 25% the reduction of MT content, protein expression decrease) and 10 fold (mean 90 % decrease) and mRNA expression in colon tissues as respectively as compared to controls treated with compared to normal tissue. This observation saline. Rats administered only saline showed 95% emphasizes the clinical relevance of this immunopositivity of the MT protein expression. multifunctional metalloprotein in colorectal However the administration of DMH resulted in carcinogenesis. We suggest that increased MT only a mean 20% MT immunopositive staining in mRNA expression and immunopositive staining the colonic tissue. The DMH treated rats when may be related to the protection against colon co­treated with aspirin or vitamin C in the carcinogenesis provided by administration of precancerous and cancerous group, the colonic aspirin, vitamin C or zinc. Abstract No: 2.9

A Study of Serum Ferritin Levels among Male Blood Donors

1

Ramesh Kumar1 and Dr.Shanmugam2 Technical Supervisor, 2Director ­ Blood Bank Services, Life Line Blood Bank & Research Centre, Tirunelveli, Tamil Nadu

Objective: Iron deficiency is the commonest cause of anemia worldwide and healthy blood donors are estimated to lose about 175 to 225 mg of iron with each donation. The objective of this study was to determine the serum ferritin levels among first time and regular male blood donors,

and also to correlate the serum ferritin levels with the number of donations and hemoglobin levels. Methods: Hemoglobin levels and serum ferritin were measured in four groups of donors divided into I group I (single time donor/year), Group II

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(twice/year), Group III (thrice/ year) and Group IV (4 times/ year). Results: The mean haemoglobin in the first time donors, II group, III group & IV group were 13.99±1.12, 13.78±1.08, 14.01±1.44 and 13.76±1.14 respectively. The changes in haemoglobin were not statically significant. The serum ferritin levels were found to be significantly lower among the regular donors ie Group IV (52.0±19.78 ng/ml) compared to first time donors (129.7±36.66), Group II (112.62±34.89) and Abstract No: 2.10

Group III (77.12±26.12). The serum ferritin levels gradually decrease according to the number of donations and there was a significant correlation between frequency of donations and the serum ferritin level. Conclusion: This date suggest that the blood banks must check serum ferritin for all the regular donors apart form haemoglobin and to advice donors about iron supplementation accordingly. This is the responsibility of blood bank towards these very important persons who are donating life to others by donating blood.

Study of Serum Magnesium Levels in Diabetic Nephropahty

M. Ravi kanth1, Dr. A.K. Bansal2 and Mrs. Lincy Jacob3 1 Demonstrator, 2 Professor, 3 Asst. Prof., Dept. of Biochemistry, Govt. Medical College, Jagdalpur, Bastar ­ 494001, Chhattisgarh. Email: [email protected] Objective: Diabetes mellitus has become a global problem with approximately 221 million people world wide, and the metabolic disregulations associated with it causes secondary pathophysiological changes in multiple organ system. Kidney is one of the mostly affected organ which leads to nephropathy. Diabetic nephropathy has become the leading cause of End Stage Renal Disease (ESRD), in developing and developed countries. Cellular magnesium is a critical cofactor in the activity of various enzymes involved in glucose oxidation like glucokinase, phosphofructokinase, enolase and also plays a role in release of insulin. Magnesium deficiency has been proposed as a novel factor indicated in the pathogenesis of diabetic complications. Significant alterations were observed in the serum magnesium levels of diabetic nephropathy patients. Methods: The present study was carried out in the Dept. of General medicine in NRI Hospital,

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Chinakakani, Gnt, D.t, A.P. from Jan ­ May 2008. The study comprised of 80 subjects, out of which 40 were cases, specifically patients of diabetic nephropathy with high serum glucose, HbA1c and urinary microalbumin and 40 were age matched controls. The statistical data was evaluated using the Z-test. Drastically increased levels of HbA1c and urinary microalbumin levels are observed in cases compared to controls. Results: The mean ± S.D, serum magnesium levels (mEq/L) in diabetic nephropathy cases were 1.92 ± 0.28 compared with controls 2.142 ± 0.16. The difference is statistically decreased (P<0.005) serum magnesium levels in cases and the Z value is 5.5. Conclusion: Thus the study concluded the assessment of decreased magnesium levels indicates the progression of diabetic nephropathy.

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Abstract No: 2.11

Effect of Aluminum on Cardiac Rhythm of Male Albino Rats: An Electrocardiogram Study

Sadhana Verma1, 2, Pradeep Kumar1, M. Y. Khan2, Sandeep Tripathi2 and A. A. Mahdi2 Departments of 1Physiology and 3Biochemistry, CSM Medical University, 2Department of Biotechnology, BBA University, Lucknow, India Objective: To date, information on the interaction of aluminum (Al) with cardiovascular functioning was limited and unclear. Al+3 is a non-essential element and known to be an extensively used in modern daily life. The aim of the present study was to evaluate the long term Al exposure on Electrocardiogram (ECG) and its correlation with oxidative markers and pathophysiology of cardiac muscle. Results: The result of `in vivo' ECG was found to be significant (p>0.05) changes in term of heart rate as compared with the age matched control rats. Increased lipid peroxide levels were markedly increased along with decreased antioxidant profiles i e., SOD, CAT and GSH in Al treated rats when compared with the controls. The electron microscopic study exhibited altered and dilated muscle fiber in Al treated rats. On the basis of the present study, Al has been demonstrated as a proMethods: We have orally administered 100 mg oxidant and its directly correlated with the altered AlCl3 (by gavage) to male albino rats (n=6) for 90 biochemical and ultra-structural profiles of the days. The same number of age and sex matched heart. rats were concurrently administered equal volume of physiological saline. After 90 days rats were Conclusion: It may be concluded that the Al performed to ECG for the evaluation of influence on the rhythm of the heart activity cardiovascular impairment. Thereafter, rats were associated with the altered antioxidant balance. sacrificed by cervical dislocation and heart was removed for the carrying out of lipid and their peroxidative product. Abstract No: 2.12

Serum Calcium, Phosphorus and Alkaline Phosphatase Levels in Preterm Babies

Sarika Singh Chauhan1, B. K. Agrawal1 and Rashmi Dwivedi2 1 Department of Medical Biochemistry, 2Department of Paediatrics, Gandhi Medical College, Bhopal. Objective: Preterm is defined as a baby with a gestation of less than 37 completed weeks. Preterms have low serum calcium and phosphorus levels with increased serum alkaline phosphatase activity. In this study serum calcium, phosphorus and ALP activity were measured in preterm babies. Methods: The present study comprised of 75 preterm babies. Of which 25 were of 28-30 weeks, 25 were of 30-32 weeks & remaining 25 were of 34-36 weeks (Controls) of gestational age. In these patients following investigations were done to assay serum calcium, phosphorus levels and serum alkaline phosphatase activity.

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1. Serum calcium by OCPC method. 2. Serum phosphorus by Modified Metol method. 3. Serum alkaline phosphatase by Kinetic p-NPP method.

Results: Serum calcium & phosphorus levels were found to be significantly decreased and Serum ALP activity was found to be significantly increased (p<0.001) at 28-30 weeks as compared Conclusion: It can be concluded that high serum to 34-36 weeks (Controls), but serum calcium & ALP activity & low serum calcium & phosphorus phosphorus levels were found to be insignificantly levels are associated with preterm babies. decreased whereas serum ALP activity were found Abstract No: 2.13

to be insignificantly increased at 28-30 weeks as compared to 30-32 weeks of gestational age in preterm babies. The correlation between serum calcium & phosphorus were found to be positive at all gestational ages, whereas Serum ALP is inversely correlated with serum calcium & phosphorus at all gestational ages.

Hypomagneseamia in Diabetes Mellitus

Shymala Manga, Dr.B.Prabhakar Rao and Dr.Sunanda

Department of Biochemistry and General Medicine, Prathima Institute of Medical Sciences, Nagunoor, Karimnagar, Andhra Pradesh, INDIA. E-mail Id: [email protected]

In the present study 20 normals as group -1 and 80 cases of diabetic patients choosen as group11. In both groups serum glucose levels and serum magnesium levels were analyzed by colorimetric method. Mean plasma Magnesium was sigAbstract No: 2.14

nificantly lower in diabetic patients (0.79 + 0.11) than control subject (1.81 + 0.56). Plasma Magnesium Correlated well both. In conculsion hypomagneseamia may predispose to Ischemic heart diseases.

Study of Biochemical Markers in Iron Deficiency Anemia

Smita B. Pawar Lecturer,Department of Biochemistry, M.V.P.'s Medical College Hospital & Research Centre, Nashik, Maharashtra. Anemia is the single most common disorder affecting mankind seen in all parts of the world, developed as well as developing countries. In India nearly 70% people are estimated to be iron deficient. It is misleading to think of anemia as a single disease with a single cause. The studies of experimental and human evidences support clear

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roles for increased oxidative stress due to susceptibility of lipids of red cell membrane to peroxidation. A low serum iron alone does not diagnose iron deficiency, a combination of serum iron, Total Iron Binding Capacity (TIBC), along with hemoglobin (Hb) and the trace element zinc (Zn) are conjunctively essential to diagnose. The

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normal adult erythrocytes can resist oxidative stress by several antioxidant defense systems. Whereas, the imbalance between oxidative stress and antioxidant system is present in iron deficient patients. In light of the several epidemiological, clinical and laboratory investigations supporting the role of oxidative stress in anemia, we aimed Abstract No: 2.15

to study the alterations in the levels of oxidants and antioxidants in-patients of iron deficiency anemia. In abridgement, present study reveals alterations in the levels of iron, Hb and TIBC, along with the augmented free radicals and impaired antioxidant potential.

Estimation of Toxic Lead (Pb) Contamination of Tapioca (Manihot esculenta) in Kerala State

Subin Joy, Shaikh Mohammed and V V Pillay Poison Control Centre, Dept of Analytical Toxicology, Amrita Institute of Medical Sciences, Cochin, Kerala 68206, Email [email protected] Objective: There have been reports of edible vegetables containing trace amounts of heavy metals above the normal permissible levels from various parts of India, including some parts of Kerala. Such contamination can have far-reaching consequences on the human population, making them potential victims of chronic heavy metal poisoning. Prolonged human consumption of unsafe concentration of heavy metals could lead to disruption of numerous biological and biochemical processes in the human body. Toxic concentration of heavy metals in the body can either act as carcinogens, or cause developmental abnormalities in children. particular reference to lead (Pb) in tapioca or cassava (Manihot esculenta) by lead dithizone colour test (qualitative), and UV/VIS spectrophotometer (quantitative). Relatively high concentration of lead in tapioca cultivated near the industrial belt of Cochin was observed, which could be due to the prevalent practice of irrigation using waste water, containing effluents from industries.

Result: Lead contamination can occur from lead pipes and lead-based solder pipe joints through water. Apart from the industrial area, the lead concentration of tapioca cultivated elsewhere was Method: This project was done to estimate the below the permissible limit of 0.3µg. content of heavy metal contamination with Abstract No: 2.16

The Effect of Vitamin E Supplementation on Soluble P-Selectin Levels in PreEclampsia

Sujata Wangkheimayum1, Sunil Kumar1 and Vanita Suri2 1 Department of Biochemistry; 2 Department of Obstetrics and Gynaecology, Postgraduate Institute of Medical Education and Research, Chandigarh. E-mail: [email protected] Background: Pre-eclampsia, a syndrome peculiar endothelial dysfunction and vasoconstriction to pregnancy is characterized by generalized manifested as proteinuria and increased blood

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pressure. P selectin, a cell adhesion molecule is elevated in many inflammatory conditions including pre-eclampsia. The effect of vitamin E supplementation on levels of soluble P-selection in pre-eclampsia has been studied here. Methods: Soluble P-selectin was measured by commercially available ELISA kit. Vitamin-E levels in plasma were estimated spectrofluorometrically. Serum and urinary creatinine as well as urinary protein were estimated by commercial kits. Results: In our study the effect of vitamin E (tocopheryl acetate) supplementation of 400 IU/ day to patients of pre-eclampsia showed Abstract No: 2.17

significant decreased levels of soluble P-selectin by the 2nd week as compared to patients given placebo (p = 0.005). Vitamin E group of preeclampsia patients had baseline median P-selectin level of 379 ng/ml, whereas, that of patients receiving placebo was 363 ng/ml. By the end of 2nd week the values were 302 ng/ml and 380 ng/ ml respectively in both the groups. Conclusion: Supplementation of 400 IU/day of vitamin E for two weeks to patients with preeclampsia showed a significant decline in levels of soluble P-selectin, a marker of inflammation. In this short period of study no direct correlations were observed between P-selectin levels and blood pressure as well as with proteinuria.

5-Methyl Tetrahydrofolate in Dried Blood Spot as an Indicator of Folate Status in Humans

Thippeswamy.T.G., Ravinder Punjal and Madhavan Nair.K Micronutrient Research Division, National Institute of Nutrition, Jamai-Osmania P.O., Hyderbad-500 007, Andhra Pradesh, India, Email: [email protected] Objective: Folic acid is a one-carbon donor essential for growth and reproduction. Its deficiency has not been mapped in India due to lack of field friendly methods of blood collection and estimation in the laboratory. Conventional assays like microbiological and competitive protein binding assay kits are sensitive but require venous blood sample. In erythrocytes, folate exists mainly as 5-methyltetrahydrofolate polyglutamates and therefore, the objective was to develop a HPLC method for estimating 5Methyl tetrahydrofolate (5-MTHF) in dried blood spots (DBS) and to establish as an indicator of folate status. Methods: Briefly, the methodology involved quantitation of 5-MTHF in venous, DBS blood

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samples and in plasma by HPLC. Stability of 5MTHF in DBS was also tested periodically. Results: The results indicated that a minimum of three DBS punches equivalent to 20µl of blood was required for the analysis. The minimum detection limit of 5-MTHF was 20 pg .The recovery of added 5MTHF in the DBS sample was more than 70%. The inter and intra-assay variations were found to be less than 10%. Conclusion: Although there was a good correlation (r2 = 0.87) between the DBS and blood 5-MTHF levels estimated by HPLC and folic acid estimated by commercial kit, folic acid levels were always higher compared to 5-MTHF. The relation ship between 5MTHF in DBS and folate status is being evaluated.

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Abstract No: 2.18

Age Related Changes in Clcium, Phosphorous and Calcitonin in Healthy Women with special reference to Premenopausal and Post menopausal Women

Miss V. Meenakshi, Dr.S.K. Sen and Dr. Roma Rattan Background and objective: The data regarding the effect of age on biochemical variables relating to bone and mineral metabolism in healthy women is inadequate. This leads to great hurdle for initiating preventive measures in age related osteoporosis in women. Though studies have examined the role of serum calcium level in osteoporosis in women, the role of age on serum calcium and anti resorptive hormone calcitonin in pre and postmenopausal women has not been elucidated. Hence the study was designed to evaluate serum calcium, phosphorous and calcitonin in the healthy women. Results: Serum calcium and calcitonin decreased significantly as compared with control group (p<0.001). Significant positive correlations were found between serum calcium and calcitonin (r=0.887, p<0.01). Serum calcitonin showed a negative correlation with serum phosphorous (r= -0.569, p<0.01). Similarly strong negative correlation was found between serum calcium and phosphorous(r=-0.603, p<0.01). Conclusion: Serum calcium and calcitonin starts to decrease in women with age greater than 40yrs

and plunges to low level in post menopausal and Methods: The study included healthy women senile women. These findings will help in initiate (n=161) aged 30-90 years. Serum calcium and phosphorous were measured in Dade Behring auto the measures in women at an earlier age to prevent analyzer. Serum calcitonin was estimated by ELISA. osteoporosis. Abstract No: 2.19

Effects of Resveratrol on Expression of Critical Bone Remodelling Factors in Surgical Menopause Model of Rat

Alka Khera, Dr. Thungapathra M, Dr. Jaswinder Kalra and Dr. Sujata Ghosh Objective: To investigate the molecular mechanisms of protection conferred by resveratrol against estrogen deficiency induced osteoporosis, using surgical menopause model of rat. Methods: The study was conducted on three months old Sprague Dawley (SD) rats which were given resveratrol (0.7 and 5mg per Kg diet) for 1 month. The rats were randomly divided into six study groups each comprising 7 animals each. (i) sham operated control group(ii) ovariectomized (iii) ovariectomized treated with á -estradiol (iv) sham operated treated with resveratrol (0.7mg/Kg diet) (v) ovariectomised treated with resveratrol (0.7 mg/Kg diet) (vi) ovariectomised treated with resveratrol (5 mg/Kg diet).At the end of 1 month , blood was collected to estimate serum osteoclastic

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and osteoblastic parameters. Femur was also removed and its proximal end was used to extract RNA and protein. Quantitative analysis of estrogen receptor á vitamin D receptor (VDR), parathyroid hormone receptor was done at gene and protein levels in all the groups.

animals we observed an increase in the expression of ER, ER and VDR in resveratrol treated animals. The expression levels of PTHR which is an osteoclastic factor, decreased in experimental animals as compared to control animals.

Conclusion: Resveratrol has protective role in Results: Concomitant with the decrease in the curing osteoporosis caused by estrogen deficiency levels of osteoporotic markers in resveratrol by directly affecting the expression of critical treated animals as compared to ovariectomised factors involved in bone remodelling. Abstract No: 2.20

An Experimental Evaluation of Cytoprotective Effect of Amifostine on CHOP Chemotherapy in Albino Rats

Ambili Remesh Dept. of Pharmacology & Therapeutics, Azeezia Medical College, Kollam; E-mail: [email protected] Background: Chemotherapeutic agents are in combination and in conjunction with surgery, radiotherapy and immunotherapy are used widely for the treatment of variety of neoplastic diseases. Drug toxicity to normal organs is a major obstacle which prevent the use of optimal doses and schedules.. Cytoprotective agents like Amifostine constitute a novel group of agents which when administered prior to chemotherapy protects normal tissues from the deleterious effects of chemotherapy with out attenuating the antitumour response. Objectives: The main objective of the experimental study was to assess the effect of toxicity induced by CHOP chemotherapy in albino rats and histopathologically demonstrate the protection of vital organs by Amifostine on druginduced toxicities. Methods: Normal albino rats (weighing150250gm) were randomly assigned to three groups. Group1 (control group) received no drugs. Group

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2 was given CHOP regimen (dose) and group 3 was administered Amifostine prior to CHOP chemotherapy. General signs and changes in behavioural, neurological, autonomic profile are noted. Haematological investigations were done and weight changes were noted. Then animals were undergone euthanasia under ether anaesthesia and histopathological examinations of internal organs were carried out. Results: Statistical data analyses were done for haematological investigations and weight changes. The level of significance was fixed at 5%. Conclusion: Study demonstrated that Amifostine has cytoptrotective effect in CHOP chemotherapy induced toxicity. There is a BIOCHEMICAL BASIS for this CYTOPROTECTION which is depicted in the mechanism of action of Amifostine. The present study further signifies the need for thorough knowledge of cell biochemistry in novel drug development for cytoprotection.

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Abstract No.2.21

Biochemical Changes in Acute Organophosphorus Poisoning

Indira A Hundekari, *A.N.Suryakar, Nilima N. Dongre, J G Ambekar, B.B.Devaranavadagi and D.B.Rathi. BLDEU's Shri B.M. Patil Medical College ,Bijapur, Karnataka. *V.M.Govt. Medical College ,Solapur, Maharashtra, Email ID- [email protected] Introduction: Organophosphorus poisoning is a major global health problemwith more than 200,000 deaths every year. Organophosphorus compounds are widely used worldwide in agriculture as well as in most household gardens .Unfortunately because of their easy availability and potent toxicity, there is a gradual increase in accidental poisoning and is commonly abused for suicidal purpose. Hospital and Research Centre, Bijapur.The samples were analyzed using standard methods for different biochemical parameters. Results: Progressive fall in plasma cholinesterase and increased serum malondialdehyde levels in correlation with the severity of organophophorus poisoning were observed. There was significant decrease in serum total cholesterol and potassium levels without much change in sodium level. C-reactive protein levels were increased. Serum magnesium levels were decreased in organophosphorus poisoning cases as compared to control group.

Objectives: Toxicities of organophosphorus compounds cause oxidative damage of cell membranes and also result in disturbed biochemical and physiological functions. Hence we planned to study the biochemical changes in acute organophosphorus poisoning. Conclusion: Inhibition of cholinesterase leads Methods: Blood samples were collected from to increased acetylcholine level which induces the clinically diagnosed organophosphorus oxidative damage resulting in various changes during acute poisoning subjects admitted during the year biochemical 2007-2009, in Shri B.M. Patil Medical College organophophorus poisoning. Abstract No: 2.22

Lead Toxicity Among Automobile Workers

Nilima N. Dongre*, A.N.Suryakar,**A.J.Patil,I.A.Hundekari,**J.A.Patil,J.G.Ambekar B.B.Devaranavadgi, D.B.Rathi and ***Rama Jailkhani BLDEU's Shri B.M.Patil Medical College,Bijapur,Karnataka. * Dr.V.M.Govt.Medical College,Solapur,Maharashtra. ** Krishna Institute of Medical Sciences,Karad,Maharashtra. *** SDM Medical College,Sattur Dharwad Karnataka. Email ID : [email protected] Introduction: In developing countries like India lead poisoning still occurs.Automobile workers are prone to long term lead toxicity due to their routine works such as motor vehicle assembly, spray painting, burning of petrol, welding, brazing, battery recharging or replacing and radiator repairing. Objectives: Occupational lead poisoning is well documented .No such study has been reported from this area. Hence the aim of the study is to observe the lead induced toxic effects among the automobile workers in the lead based workplaces from Bijapur city.

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Methods: A study on toxic effects of occupational lead exposue among 27 automobile workers and 25 nonexposed healthy controls from Bijapur city was carried out. Lead levels in the blood and urine were analysed by atomic absorption spectrophotometer and standard assay procedures were used for other biochemical parameters.The biochemical parameters studied were serum bilirubin,total protein,albumin, globulin,A/G ratio,gamma-glutamyl transferase, alanine transaminase,aspartate transaminase,alkaline phosphatase, urea,creatinine and uric acid. Results: The study shows that blood lead and urine Abstract No: 2.23

lead levels were significantly increased in the automobile workers. as compared to nonexposed healthy controls (P<0.0001). It is also observed that there is significant change in some liver function and kidney function tests in automobile workers. Conclusion: The study indicates lead toxicity still persists in automobile workers .The study helps to create awareness about the toxic effects of lead and may entail establishment of regulations for the precautionary measures to be taken among the lead exposed workers.

Tacrolimus in Renal Transplant Recipients

Dr.Pradeep Naik, and Mallikarjuna .M Department of Clinical Biochemistry, Global hospitals, Hyderabad, Email : [email protected]

Objective: AUC-guided dosing of tacrolimus prevents progressive systemic overexposure in renal transplant recipients. Tacrolimus has a narrow therapeutic drug and bioavailability is known to vary considerably between renal transplant recipients. Most of the transplant centers still rely on measurement of trough levels, but there are conflicting reports on the correlation between tacrolimus trough levels and systemic exposure, as measured by the area-under-theconcentration-over-time curve (AUC(0-12h)).

high concentration in 14 patients[ 99 % ] the squared correlation with the AUC (0-12h) (r 2= 0.94).the concentration of four hours post-dose blood is proportional to the increase of the drug dosage .

Results: Compared with trough level monitoring only, this approach reduced the 95%-prediction interval by 50%. The Bayesian approach proved to be feasible in clinical practice, and provided accurate information about systemic tacrolimus exposure in individual patients. In the AUC-guided Methods: We studied and analyzed in 15 renal dosing cohort the apparent clearance of tacrolimus transplant recipients - based pharmacokinetic decreased gradually over time, which was not model with ELISA use of TMB as a substrate to reflected in corresponding trough levels. estimation of tacrolimus systemic exposure.. The main objective was to study intrapatient variability Conclusion: This simple, flexible method provides in the drug concentrations in blood versus different the opportunity to predict immunosuppression, and doses. Bayesian forecasting with a two-point should help minimize tacrolimus-related toxicity, sampling strategy, a trough level, and a second such as nephrotoxicity and post-transplant diabetes sample obtained four hours post-dose significantly mellitus.

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Abstract No: 2.24

Biochemical Effects of Feeding Soft Drink and Ethanol

Arun Raj, Praveen Kottath Veetil, Sheeba Varghese, J K Mukkadan and P K Joseph Little Flower Institute of Medical Sciences and Research, Angamaly 683 572, India; Email ID: [email protected] Objectives: This work was undertaken to study whether consumption of alcoholic beverage mixed with soft drinks could reduce the metabolic effect caused by ethanol. Methods and results: When 24 hr fasted rats were intragastrically fed rum (with 40% ethanol) diluted (1:1) with water, 3.0 ml (0.5 g ethanol) per 100 g body weight and sacrificed 12 hr later in fasting condition, exhibited higher levels of triacyl glycerol, glucose, total cholesterol, high density lipoprotein (HDL), aspartate amino transferase (AST), alanine amino transferase (ALT) and alkaline phosphatase (ALP) in serum, higher levels of total cholesterol, triacyl glycerol and thiobarbituric acid reactive substances (TBARS) in both liver and kidneys, and lower levels of serum albumin. When fasted rats were fed 3.0 ml soft drink (0.31 mg caffeine), they showed Abstract No: 2.25 increased levels of triacyl glycerol, glucose, ALT and ALP in the serum, TBARS in liver and kidneys, triacyl glycerol and total cholesterol in kidneys and lower levels of serum albumin. Soft drink feeding did not reduce serum total cholesterol but reduced HDL levels. Also soft drink did not alter liver lipids. When a mixture of 1.5 ml diluted rum (0.25 g ethanol) and 1.5 ml soft drink (0.154 mg caffeine) were fed to the fasted rats, the serum parameters increased similar to rats fed rum only except that total cholesterol and HDL cholesterol were unaltered. TBARS in kidneys and liver were also increased but triacyl glycerol levels were not altered. Conclusion: Thus feeding ethanol with soft drink does not reduce the metabolic effects of ethanol but it will prevent ethanol induced serum HDL cholesterol rise.

Comparative Study for Slow Acetylator Status by Phenotype / Genotype Polymorphism of N-acetyltransferase-2 in Patients on Antitubercular Treatment

S.V.Rana, Sanjeev K.Sharma, R. P. Ola, S.Arora*, S.K.Sinha, P Pandhi** and K.Singh Departments of Gastroenterology , Immunopathology* & Pharmacology**, Postgraduate Institute of Medical Education and Research, Sector 12, Chandigarh­160012, UT, INDIA. Background: Isoniazid and rifampicine are traditionally used as first line therapy for tuberculosis. Acute hepatitis frequently develops in patients receiving these drugs. Isoniazid is the major drug incriminated in this hepatotoxicity and mainly metabolized to hepatotoxicity intermediates by N-acetyltransferase-2 (NAT2).Slow acetylators have been reported more prone of hepatotoxicity. However, acetylator status of a patient can be measured in blood phenotypically by Isoniazid load test and genotypically by NAT2 gene polymorphism. But phenotypically measurement can be affected by many other factors but easy to measure and does

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not require PCR and electrophoreses and more time.

fast acetylators measured by phenotypically using Isoniazid load test. While only 20 out of 184 (12.2%) were slow acetylators and 160 out of 184 Objective: The present study was focused to (87.8%) were fast acetylators genotypically using compare the results of both the methods in patients a PCR with RFLP of gene NAT2 polymorphism. on antitubercular treatment. These results shows that slow acetylators were Methods: One hundred eighty four adult North more when measured phenotypically as compared Indians between age 18-65 yrs were enrolled in to genotypically. this study. Measurement of slow acetylator Conclusion: This study indicates that phenotypically was done by Isoniazid load test and genotypically estimation of slow acetylator status genotypically by NAT2 gene polymorphism in in patients on antitubercular treatment using NAT2 patient's blood by PCR-RFLP method. polymorphism can not be replaced by Results: Out of 184, forty-four patients (26.8%) were slow acetylators and 140 out of 184 (73.2%) Abstract No: 2.26 phenotypically measurement of slow acetylators using isoniazid load test.

Correlation of Gall Stone Disease with Iron-Deficiency Anemia

1

Prakash A; 1Lal A.K; 2Sachan P.K and 2Sahu S.K. Department of 1Biochemistry and 2Surgery, Himalayan Institute of Medical Sciences, Jollygrant, Dehradun, Email: [email protected]

Introduction: Gallstone disease is a common clinical entity affecting the adult population of both sexes. There are many causes for the disease. The one main important cause may be the iron deficiency anemia. So our main study was to correlate iron deficiency anemia with gallstone disease. Methods: 100 patients suffering from cholelithiaris admitted in our hospital with confirmed diagnosis by ultra sonography were included in the study irrespective of their age, sex and parity. Serum Iron, serum total Cholesterol and biliary Cholesterol was estimated in all the patients. Based on serum iron content, the patients with choleslithiasis were divided into two groups. Group A contained patients with normal serum iron and Group B iron deficient patients. Serum

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and biliary cholesterol contents of both groups were analyzed and compared with each other. Results: The mean bile cholesterol level in Group A was found to be 214.6 mg/dl and in Group B it was 375.3 mg/dl. The difference in values in both the groups was found to be statistically extremely significant (P<0.0001). Iron deficiency has been shown to alter the activity of several hepatic enzymes leading to increased gall bladder cholesterol saturation and promotion of cholesterol crystal formation. Conclusion: The present study suggests that iron deficiency anemia is playing a significant role in the super saturation of gallbladder bile with cholesterol and may act as an independent factor in formation of cholesterol gallstones.

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Abstract No: 2.27

Trace Elements in Leprosy

Dr V B Londhe, Sevarina Madhale*, Dr Z G Badade**, Dr P E Jagtap***,and Dr S K Ahaley**** Assoc. Professor, Biochemistry Dept., JNMC, Belgaum, *Jr. Research Fellow, RLH, Miraj, ** Prof. & Head, Dept. of Biochemistry, MGM Medical College, Navi-Mumbai, *** Prof. & Head, Dept. of Biochemistry, Bharati Vidyapeeth's Medical College, Miraj, **** Professor of Biochemistry, Bharati Vidyapeeth's Medical College, Miraj. [email protected]

Background: Leprosy (Hansen disease) is a chronic infections disease of man caused by mycobacterium leprae. Trace elements play an active role in various metabolic processes of the body. In recent years there has been an enormous awareness and understanding of role of trace elements in health and disease. Trace elements catalyse and control biochemical reactions. More than half of the body enzymes have one or more of trace elements incorporated at their active sites, many enzymes depend upon association with trace elements for their activity.

matched healthy subjects were selected from Civil Hospital, Sangli and Richardson leprosy Hospital, Miraj. The trace elements viz. Zn, Cu, Mn, Mg, and Se were estimated by Atomic Absorption Spectophotometry, at USIC, Shivaji University, Kohlapur and vanadium was estimated by ISE method at NCL, Pune. Results: In our study as per increase in bacillary load the oxidative stress increased in different types of leprosy. Trace elements viz., Zn, Mn, Mg, Se and V decreased, while Cu increased as sialylated ceruloplasmin increased in peripheral blood.

Objective: The aim of our study was to explore Conclusion: The trace elements zinc, copper, the role of trace elements in the leprosy. manganese, magnesium, selenium and vanadium Methods: In present study fifty clinically may act as biochemical markers in diagnosis, diagnosed leprosy patients and fifty age and sex differentiation and prognosis of leprosy. Abstract No: 2.28

Oxidative Stress, Antioxidants and Trace Elements in Leprosy

Dr V B Londhe, Sevarina Madhale*, Dr Z G Badade**, Dr P E Jagtap***and Dr S K Ahaley**** Assoc. Professor, Biochemistry Dept., JNMC, Belgaum, *Jr. Research Fellow, RLH, Miraj, ** Prof. & Head, Dept. of Biochemistry, MGM Medical College, Navi-Mumbai, *** Prof. & Head, Dept. of Biochemistry, Bharati Vidyapeeth's Medical College, Miraj, **** Professor of Biochemistry, Bharati Vidyapeeth's Medical College, Miraj. [email protected] World wide leprosy continuous to be a major public problem. Leprosy is an infectious disease and occurs principally in developing countries. Mycobacterium leprae is a causative organism of leprosy. At present, 1.3 million people are suffering from leprosy. Less attention has been paid towards free radical, antioxidant and trace elements study in leprosy. Considering these facts our study was

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aimed to find out correlation between oxidative stress, antioxidants enzymes and trace elements in leprosy. In present study fifty clinically diagnosed leprosy patients and fifty age, sex matched healthy subjects were selected from Civil Hospital, Sangli and Richardson Leprosy Hospital, Miraj. Serum lipid perode, SOD, Catalase and glutathione peroxidase were estimated by spectrophotometric methods. The trace elements viz. Zn, Cu, Mn, Mg, and Se were estimated by Atomic Absorption Spectophotometry, at USIC, Shivaji University, Kohlapur and vanadium was estimated by ISE method at NCL, Pune. In our study, with

extent of disease progression, the oxidative stress increased, due to increase in the bacillary load. For scavenging purpose, the trace elements Zn, Mn, Mg, Se and V with their respective antioxidant enzymes SOD, GPx and Catalase decreased. Antioxidant enzymes and trace elements showed negative correlation with lipid peroxide and bacillary load. There was positive correlation between copper and lipid peroxide with bacillary index. The estimation of LPO, SOD, GPX, catalase and trace elements serum Zn, Cu, Mn, Mg, Se and V as biochemical markers for diagnosis and prognosis of leprosy.

Abstract No: 2.29

Variations in Serum Ferritin, Electrolytes and TIBC Levels in Paediatric Multitransfussed Thalassemic Patients

Jaidev Singh, Dr. (Mrs) Meena Varma and Dr.Sangeeta Paneri M.G.M Medical College Indore (M.P). Email ID: [email protected] Introduction: Beta thalassemia is a single gene disorder requiring regular multi-blood transfusion which causes serious side effects an overload of iron in the form of ferritin.Excess or free iron can catalyze the formation of very toxic compound such as OH- radicals through Fenton reaction which causes oxidative damage. This oxidative damage causes the pathogenesis of the membrane abnormalities observed in beta thalassemia.. Objective: We determined whether thalassemia could account for abnormal cation transport system. Traditionally serum iron, ferritin, TIBC and the % saturation, an indirect measurement of transferrin, have been used to determine iron status. Methods: In the present study included 75 multitransfused thalassemic children and 35

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healthy control.All were in the age group of 3-10 years. And evaluated the serum ferritin by ELISA METHOD, electrolytes by Flame photometry, TIBC by RAMSAY DIPYRIDYL method Results: In the present work it was found that, the significantly increased serum levels of ferritin while normal mean levels of TIBC in thalassemia patients as control group. The serum Na+ & K+ levels were found to be significantly decreased in thalassemic patients respectively. Conclusion: It can be concluded that the increased & decreased or normal TIBC are related to thalassemia. And defective membrane transport is responsible for observed changes of electrolytes homeostasis in thalassemia.

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Abstract No: 2.30

Study of Lead Content in Various Water Samples within Bangalore and A Survey Based on Effects of Lead Poisoning on People

Preeti Mangala*, Javeria Anjum and Arpi Dey *Department of Chemistry, Jyoti Nivas College (Autonomous), Bangalore-560095, India, email: [email protected] (Dr. Preeti Mangala, Lecturer in Chemistry, Jyoti Nivas College (Autonomous), Bangalore, Ms. Javeria Anjum, Student of III B.Sc. (Biotech, Botany, Chemistry), Jyoti Nivas College, Bangalore, Ms. Arpi Dey, Student of III B.Sc. (Biotech, Botany, Chemistry), Jyoti Nivas College, Bangalore) Objective: Water is an essential component for survival of life on earth. We require water for each and every activity that we do. The source of lead in water can be domestic, environmental and occupational. Keeping in mind the above sources, it was considered worthwhile to investigate few water samples from residential areas like Koramangala and BTM Layout (near to our college), lakes such as Ulsoor, Madiwala, Agara etc. and area where there is battery recycle units such as Tilaknagar for the lead content. Methods: The estimation of lead content in about 41 water samples was done by Graphite Furnace Atomic Absorption Spectrometry (GFAAS) method followed by comparative studies. For residential areas, the comparison was between source (treated water/Cauvery water) and kitchen based on plumbic and non-plumbic supply of Cauvery water, mixing of Cauvery water with bore- well water etc. For lake water, the influential factors were disposal of wastes, immersion of painted Ganesha idols if any and rain. The amount of lead was relatively high in the water samples contaminated through occupational sources as expected. Results: The result shows that bore-well water of Koramangala (especially 7th & 8th Block) has more lead content than Cauvery water. This may be due to presence of printing press, automotive repair shops, industrial machinery etc. around or plumbic fixtures or fittings in those residences. Further, mixing of Cauvery water to bore-well water reduces the lead content. The relatively greater lead content of Koramangala water tank may be due to degrading PVC pipes, hard plaster, lead paints, tile & brick glaze, seals for water mains etc. The experimental data proves that the borewell and some of the well water are contaminated with a very little amount of lead in the residential area of BTM Layout which may be due to construction of building/flyover, demolition of old houses, painting, plumbic fixtures and fittings, pottery, industrial machinery work etc. It is noteworthy to mention that the lead content in various water samples of Cauvery water reported in this project were found to be appreciably small and it can be regarded as `Lead-safe water'. The significant decrease of lead content in the water of Ulsoor, Madiwala and Agara lakes from May 2008 to September 2008 may be due to dilution as a result of heavy rain. This also attributes to the ban of immersing the painted Ganesha idols in the lakes of Bangalore city after the holy festival of `Ganesha Chaturthi' and promoting the eco-Ganesha which is a great achievement of the entire team of National Referral Centre for Lead Poisoning in India (NRCLPI) and Quality Council of India (QCI) for conducting several awareness programmes/workshops related to prevention of lead poisoning in various schools/colleges within the city. The relatively high lead contamination of

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Tilaknagar well water reported here may be due of Biochemistry, St. John's National Academy of to occupational lead exposure like breaking, Health Sciences, Bangalore for awarding this short recycling and manufacturing batteries. term research project; to Dr. S. Muralidhar Rao, Sr. Manager ­ Technical Operations, Conclusion: The overall result was found to be Aquadiagnostics (AWRTCL), Bangalore in within desirable maximum limit according to analysing the lead content by GFAAS method; to W.H.O. (50 ppb or ìg/L), but we must further Dr. P.A. Shankar, Director, Filtrex Technologies alleviate lead for "lead-safe water". Pvt. Ltd., Bangalore for discussion on research Acknowledgement: We are thankful to Dr. methodology, and to Sr. Philomena Cardoza, Thuppil Venkatesh, Director (NRCLPI, Advisor Principal. Jyoti Nivas College (Autonomous), of Quality Council of India (QCI), Professor, Dept. Bangalore for her support and financial assistance. Abstract No: 2.31

Evaluation of Lead Concentration in Milk and Milk Products

Charanraj T.P1*, Anuj Ranjan, Alka Kumari, Arun Kumar, Kalpana Devi.N and Sushma Bharti 1.Department of Biochemistry, Reva Institute of Science and Management. Bangalore. Email: [email protected] Twenty five milk samples were collected from different places using acid treated Lead free polyethylene bottles and evaluated for the concentration of Lead using AOAC method of extraction and assayed by Graphite furnace atomic absorption spectrophotometer (GFAAS).The data showed that Lead is present in diversified conAbstract No: 2.32 centrations among the milk samples obtained from different regions. The current study revealed that the Lead concentration in some places of study is above the WHO limits (0.02ppm or 20 ppb).The current study was conducted during monsoon. Seasonal fluctuations if applicable have not been included

Effect of Lead on Employees Working in Paint Industry

Hariprasad.B.N, Vinay.G, Vinay Kumar.C, Sathish Kumar.P, V and Prof. Sripathy.L HOD of Chemistry; Vivekananda Degree College, Department Of Chemistry, Dr. Rajkumar Road, Rajajinagar, Bangalore-55 Under the guideines: Dr. Venkatesh Thuppil, Principal advisor QCI & NRCLPI Awareness about the toxic effects of heavy metals (LEAD) is still lacking in developing countries. Lead is one among them, which ranks second in the Agency for Toxic Substances and Disease Registry's top 20 lists of toxic metals. There is still one source of lead regularly causing poisoning, namely paint, on which this paper focuses.

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Lead poisoning has been known for centuries. Childhood poisoning from lead­based paints was first noted in 1897 in Australia, where children ate paint chips from porch railings this prompted the City of Brisbane to pass specific legislation designed to prevent poisoning from lead­based paint. Lead concentration of blood samples from paint

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industries were determined by meta-exchange reagent followed by Anodic Stripping Voltammetry method. The main aim of this study is to determine the blood lead level of the employees working in Paint industry, from in and around Peenya industrial area (Bengaluru), to asses the feasibility of using the lead in paints. Some of the paints being used even now are known to contain this toxic metal at alarming levels. We have a list of persons who are quite often suffered from headaches and back aches. Lead is mainly used in

paints for speeding up its drying, & for giving them desired colors. A wide range of colors contain lead, for instance, Yellow contains Lead (II) chromate i.e. PbCrO4, White color contains lead (II) carbonate, PbCO3. The Blood lead level is ranging from 9.4 to 26.7. Correlating with the values of control it was found that there is a considerable rise in the blood lead level. Concluding, the employees in the paint industry have the Blood lead level at alarming levels.

Abstract No: 2.33

Vitamin D- Emerging Silent Lifestye Disease: A Pilot Study

D. Radhika Chowdary and B. Naveen Kumar Department of Laboratory Medicine, Krishna Institute of Medical Sciences; Secunderabad-500003

Aim: To estimate the 25 hydroxy vitamin D levels in patients coming to outpatient department with generalized musculo skeletal pains and muscle cramps.

by Phosphomolybdate reduction on Vitros 250 dry chemistry analyzer.

Results: Out of 128 patients 91 (67%) were Vitamin D deficient with normal calcium and Objective: To establish a relationship between phosphorus levels. Mean of vitamin D 15.07 ± 7.51 µg/dl, phosphorus 3.66 ± 0.61 mg/dl and calcium vitamin D deficiency and the generalized 9.59 ± 0.70 mg/dl in these deficient individuals. symptoms which were not due to any other specific cause for which they were investigated. Discussion: There was deficiency of vitamin D in The deficiency of vitamin D leads to bone pains 67 % of the patients with normal Calcium & & fractures, muscle cramps and in severe cases Phosphorus levels. The deficiency seems more osteomalacia. The threshold for optimum Vitamin prevalent than thought in tropical country like India D status is known to be considerably higher than where sunlight is adequate. Deficiency may be due to too little time spent outdoors & tendency to that required to prevent osteomalacia. switch to vegetarian diet where Vitamin D is very Methods: Random venous samples of 128 low. Detection of Vitamin D levels and treatment patients were taken from Jan to June 2009 for with it alleviates most of the symptoms. However estimation of Vitamin D,calcium & phosphorus this is only a pilot study with patients coming to levels. Vitamin D was estimated on Elecyses our hospital. We need to take up different Roche 2010 by electro chemiluminescence. population study based on activity, socioeconomic status & lifestyle for more detailed data. Calcium was estimated by Arsenazo, phosphorus

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Abstract No: 3.1

Myeloperoxidase in Chronic Kidney Disease

Usha Anand, Madhusudhana Rao.A, Vijaya.D, Aruna V, Gayathri.B and Anand C.V Department of Biochemistry, PSG Institute of Medical Sciences & Research, Coimbatore-641004. Email: [email protected] Background: Myeloperoxidase (MPO), a prooxidant enzyme is released from granules of activated neutrophils at inflammatory sites. High levels of MPO have been implicated in the pathogenesis of CVD. As chronic kidney disease (CKD) is a risk factor for CVD, we attempted to estimate levels of MPO in patients with CKD including end-stage renal disease (ESRD). Methods: Subjects of this study were classified into three groups based on GFR which was calculated using MDRD formula. Group I consisted of control subjects (n=20) with normal kidney function and a GFR > 90 ml/min/1.73m2. Group II and III included patients with CKD. Group II (n=30) had GFR between 15 and 89 ml/ min/1.73m2 while group III (n=25) had ESRD and GFR < 15 ml/min/1.73m2. Plasma MPO was assayed by a spectrophotometric method. Serum urea and creatinine were estimated on a clinical Abstract No: 3.2 chemistry analyzer using standard laboratory procedures. Results: Plasma MPO levels progressively lowered with advancing renal failure. Plasma MPO levels (U/L) were 62.07 ± 19.76, 46.68 ± 20.03 and 28.73 ± 12.50 (mean ± SD) in groups I, II and III respectively. MPO levels were significantly lower in groups II and III when compared with group I (p<0.05). There was a significant positive correlation between GFR and MPO (r = +0.458, p<0.001) and a significant negative correlation with urea (r = -0.410, p<0.01) and creatinine (r = 0.456, p<0.001). Conclusion: Contrary to assumption, MPO levels decline with progression of CKD. Decline in plasma MPO levels may be due to the inhibitory effect of uremic toxins on the enzyme.

Serum Adenosine Deaminase Levels in Acute Kidney Injury

Saleena Ummer V1, Nalini K2 , Ravindra Prabhu3 and Licy CD1 1 Dept. of MLT, MCOAHS-Manipal, 2Dept. of Biochemistry, 3Dept. of Nephrology, Kasturba Medical College, Manipal University Background: Adenosine deaminase (ADA) is a non-specific marker of the T cell activationand hence important in acute and protracted inflammatory responses. Inhibitors of ADA can attenuate the elevation of pro-inflammatory

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factors. However the effect of ADA in renal dysfunction has not been adequately studied. Objective: The purpose of this study was to correlate the activity of serum ADA in patients with

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acute kidney injury (AKI) and to establish a group .ie, 63.41±11.56 IU/L vs 23.39 ± 5.22 IU/L relationship between ADA activity and AKI if any. with a p value of <0.001. Any correlation between serum ADA activity and creatinine level could not Methods: Serum ADA activity was determined be established. in 28 cases with AKI. These patients were diagnosed with sepsis, leptospirosis and snake Discussion: Although serum levels of ADA do not bites. 25 healthy individuals served as controls. correlate with that of creatinine, its serum level is Serum creatinine levels were estimated (by Jaffe's increased in AKI with a higher positive predictive method) to confirm the kidney injury. ADA value. Many effects produced by ADA are caused activity was determined by Guisti and Glanti's by the metabolism of adenosine. It is known that (colorimetric) method. The cut off value was taken adenosine can attenuate the ischemic/reperfusion as 35 IU/L. The ADA activity was compared in injury. If adenosine is rapidly metabolized by the patients with AKI with that of healthy controls. high level of ADA, the beneficial effects of Correlation analysis between serum ADA activity adenosine will be lost. This would result in the and creatinine level was also performed. production of superoxide radicals thereby exaggerating kidney injury. However to study the Results: Out of the total 28 cases of AKI, 17 precise mechanisms involved, further showed an elevated serum ADA activity. Mean characterization of the enzyme and determination serum ADA levels were significantly higher in of its cellular source is essential. those with acute kidney injury than in the control Abstract No: 3.3

In vitro Studies on Anticalcifying Properties of achyranthes aspera

Aggarwal A*, Singla SK** and Tandon C* * Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan ­ 173215, **Department of Biochemistry, Panjab University, Chandigarh Objective: Kidney stone is one of the oldest and most widespread diseases known to man. Calcium containing stones, especially calcium oxalate monohydrate (whewellite), calcium oxalate dihydrate (weddellite) and calcium phosphate (apatite) are the most commonly occurring ones. The present study is aimed at examining the efficacy of Achyranthes aspera on mineralization, growth and demineralization of Calcium Phosphate in vitro. to study its anticalcifying properties. For initial mineralization of calcium phosphate, a 5ml assay system containing 0.05M Tris HCl (pH 7.4), 5mM CaCl2, 5mM KH2PO4 and aqueous extract was prepared. After incubation and centrifugation, the pellet was dissolved in 0.1N HCl for estimation of calcium and phosphate.

Results: The growth and demineralization assays were performed using the preformed mineral phase. Calcium and phosphate estimations were Methods: The roots of Achyranthes aspera were performed in the pellet and the supernatant for the powdered and the aqueous extract was prepared growth and demineralization assays respectively. by boiling the sample, and then filtering it. This Achyranthes aspera was found to inhibit aqueous extract obtained after filtering was then mineralization and demineralization of calcium used as a test sample for the various assay systems and phosphate in vitro in a concentration dependent

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manner. Aqueous extract of Achyranthes aspera also inhibited the growth of calcium and phosphate on preformed mineral phase.

Conclusion: Based on experimentally demonstrated properties, Achyranthes aspera is a valuable candidate for developing new phytotherapeutic agents.

Abstract No: 3.4

Correlation of Serum Intact Parathormone Level with Biochemical Parameters in Various Stages of Chronic Renal Failure Pateints

Bhimappa N.M, Suma M.N and H.S Virupaksha E-mail: [email protected] Background: The secondary hyperparathyroidism is a common, important and treatable complication of chronic renal insufficiency (CRI) and end-stage renal disease (ESRD). Although its exact pathogenesis is not known, hyperphosphatemia, hypocalcemia, deficiency of vitamin D, decreased expression of calcium and vitamin D receptors, and parathyroid hormone (PTH) resistance have individual roles. Longstanding secondary hyperparathyroidism results in osteitis fibrosa cystica (a high turnover bone lesion) and increases the risks for bone pain and fractures. Objectives: 1.To estimate serum intact parathormone and other biochemical parameters like serum creatinine, blood urea, serum calcium and inorganic phosphate in chronic renal failure patients and healthy volunteers. 2.To compare serum intact parathormone level with biochemical parameters in chronic renal failure patients and healthy volunteers. 3.To find if there is any correlation between serum intact parathormone level and biochemical parameters in the study group. Methods: 30 patients in each stage of chronic

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renal failure (totally 150 CRF patients) and 30 healthy controls in the age group of 20-60 years were taken for the study. Serum intact parathormone (iPTH) was estimated by chemilumino-metric assay, serum total calcium by Arsenazo-III method, serum inorganic phosphate by ammonium molybdate method, blood urea by urease method and serum creatinine by Jaffe's method. Creatinine clearance was calculated using Cockcroft and Gault equation. Results: The level of serum iPTH was significantly higher in more advanced renal failure, thus confirming the relationship between severity of hyperparathyroidism and the degree of renal impairment. The increased levels of mean serum iPTH were present even in early renal failure, and it was related to low mean serum calcium level and progressive rise of serum inorganic phosphate from early to advanced renal failure. So serum iPTH is negatively correlated with creatinine clearance (GFR) and serum total calcium. However, serum iPTH is positively correlated with inorganic phosphate, urea and creatinine. Thus progressive hypocalcemia and hyperphosphatemia are the initiating factors for the development of secondary hyperparathyroidism in CRF patients.

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Abstract No: 3.5

Comparison of Serum Creatinine, Urinary Creatinine and Urinary Beta 2 Microglobulin Levels as an Early Marker of Subtle Renal Dysfunction due to Aminoglycoside Therapy in Full Term Sick Neonates.

Dr Bonny B Jasani, Dr Rohini Bhadre and Dr Varsha D. Phadke Dept of Biochemistry, Second floor, Hospital building, K.J.Somaiya Medical College and Research Centre, Eastern Highway,Everad nagar, Sion Mumbai - 400022,Maharashtra, Email Id: [email protected] Objective: Comparison of serum creatinine, urinary creatinine and urinary beta 2 microglobulin levels as an early marker of subtle renal dysfunction due to aminoglycoside therapy in full term sick neonates. Methods: In this prospective randomised case control study sixty six full term sick neonates admitted in NICU of a tertiary hospital from December 2007 to November 2008 requiring aminoglycoside treatment and thirty normal neonates matched appropriately were enrolled in the study. Apart from routine septic profile, the parameters to assess renal function included serum creatinine, urinary creatinine and urinary Beta 2 microglobulin which were estimated on day of admission and on Day 3, Day 7. The investigations done in controls were urinary Beta 2 microglobulin on Day 1 and Day3 of life. All sixty six full term sick neonates were administered Abstract No: 3.6 cefotaxime and gentamicin or amikacin alternately for duration of minimum seven days. Results: In our study, the percentage of renal dysfunction diagnosed by serum creatinine was 28% and by urinary beta 2 microglobulin was 68%. This signifies that 40% of the neonates with subtle renal dysfunction would be missed if traditional tests are used in diagnosis. It also revealed that though Beta 2 microglobinuria is an extremely sensitive indicator of proximal tubular dysfunction, it was a relatively insensitive test to diagnose aminoglycoside nephrotoxicity due to coexistence of underlying clinical conditions in sick neonates. Conclusion: Beta 2 microglobinuria is a sensitive and early indicator of proximal tubular function but its clinical utility in diagnosis of aminoglycoside nephrotoxicity is limited due to presence of multiple confounding factors.

Observations on Microalbuminuria in Hypertensives with Alcholism and / or Smoking

Dr.D.L.Lalitha1, Dr.B.Sreehari babu2, Mr.D.S.S.K.Raju2 and Dr.P.K.Behera3 1.Associate Professor, 2. Tutor, 3.Professor, Department of Biochemistry, MIMS, Nellimarla, Vizianagaram, A.P; Email ID: [email protected]

Methods: 60 cases of hypertensive patients were selected who were negative for albustix in urine and were assessed for incidence of microalbuninuria with reference to their habits of

smoking and alcoholism. Out of 60 hypertensive patients 46 (77%) were found to have microalbuminuria. Prevalence of microalbuminuria in males and females were found

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to be 77% and 75% respectively in those 46 cases. 48 normal healthy individuals who were not hypertensive and were non-alcholics and nonsmokers constituted the control. The twenty-four hours urine samples were collected and stored with preservative. The urinary microalbumin was estimated by using pyrogallol method (Pointe scientific,inc..USA) Microaluminuria was also calculated in terms of albumin/creatinine ratio (mg/g). Results: In the present study microalbuminuria in controls and hypertensives was found to be 19.02mg/day±5.25 and 77.08mg/day±37.35. The increased microalbuminuria in hypertensives was statistically significant (p<0.001). Albumin creatinine ratio in controls and cases were found to be 14.44 mg/g±4.62 and 57.62mg/g±30.66 respectively. The increase of albumin creatinine ratio in hypertensives was also found to be Abstract No: 3.7

statistically significant (p<0.001). Prevalence of microalbuminuria in nonsmokers and nonalcoholic hypertensives was found to be 14%. However the prevalence in alcoholics, smokers and both smoker & alcoholic groups were found to be 22%, 32% and 32% respectively. The mean urinary microalbumin level in smokers, alcoholics, smokers&alcoholic and nonsmoker & nonalcoholic groups were found to be 97.22mg/ day, 89.66 mg/day, 122.66 mg/day and 80.5 mg/ day respectively. The alcoholics and smokers exhibited statistically significant (p<0..001) higher level for microalbumin as compared to other groups. These findings suggest both smoking and alcholism adversely affects the renal functions. Conclusion: Thus it becomes immensely important in respect of therapeutic implementation and educating the hypertensives regarding the ill effects of smoking and alcoholism

Serum Lipoprotein(a) and Lipid Profile in Chronic Kidney Disease

Neela B M, and Virupaksha H S Department Of Biochemistry, J.S.S Medical College, Mysore, Karnataka; Email Background: Chronic kidney disease is characterized by progressive loss of renal function. Cardiovascular disease is the leading cause for morbidity and mortality in these patients. Lipoprotein(a) [Lp(a)] is a cholesterol rich particle existing in the human plasma. In chronic kidney disease patients, lipoprotein(a) level is increased. Increased lipoprotein(a) level is risk for cardiovascular disease. Objectives: 1.To estimate serum lipoprotein(a) in chronic kidney disease patients. 2.To estimate total cholesterol, HDL-Cholesterol, LDL-Cholesterol and triglyceride levels. 3. To compare the above with age and sex matched healthy controls. 4. To study if there is any correlation between

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lipoprotein(a) and cholesterol, triglyceride, LDLCholesterol and HDL-Cholesterol. Methods: The study group included 30 patients diagnosed as chronic kidney disease patients and 30 healthy controls. Serum lipoprotein(a) was estimated. Total Cholesterol, Triglyceride, LDLCholesterol, HDL-Cholesterol were correlated with lipoprotein(a) levels. Results and Discussions: There was significant increase in lipoprotein(a) level in patients with chronic kidney disease when compared to controls. Increase in lipoprotein(a) level was statistically significant. Increased Lp(a) level along with increased triglyceride, LDL-Cholesterol could lead to the accelerated tendency of atherosclerosis and other cardiovascular abnormalities in these patients.

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Abstract No: 3.8

Evaluation of Urinary Protein Bound Sialic Acid and Protein Carbonyl content as the Prognostic Markers of Nephrotic Syndrome in Pediatric Age Group

Niranjan.G, Koner B.C and Bhat .V Department of Biochemistry and pediatric , JIPMER; Email ID: [email protected]

Introduction: Nephrotic syndrome (NS) is a common disorder affecting mainly the pediatric age group, 90% of the NS cases are idiopathic which can be classified as steroid sensitive (SS), steroid dependent (SD) and steroid resistant (SR) cases depending on response to treatment with steroids. There is no way to predict at the onset which cases are going to be SS, SD or SR cases. Hence the present study was designed to find out if urinary protein bound sialic acid (UPBSA) and urine protein carbonyl content (UPCC) differ in SS, SD or SR cases of NS and also to find out estimation of these parameters before starting the treatment can predict the prognosis of these cases. Methods: 70 NS cases were enrolled in the study. UPBSA and UPCC were estimated by Abstract No: 3.9

modifications in Aminoff's and Levine'smethods respectively in all the cases before starting the steroid treatment. Later all the cases were followed up out of which 47 cases became SS also we also got 15 SD and 8 SR cases. Results: Both UPBSA and UPCC s were significantly low in SS compared to SD and SR cases. ROC showed a cutoff limit of 2.74 µgm/mg for UPBSA and 7.02 n moles/mg protein for UPCC to predict SD/SR cases of NS. Conclusion: Urinary protein bound sialic acid and urinary protein carbonyl content can be used as a marker to predict SS, SR and SD cases of NS.

Oxidative Stress and Transforming Growth Factor-Beta 1 (TGF-â1) in the Progression of Diabetic Nephropathy

Kavitha Gandhi* and Dr.Rita Mary Aruna** * Vinayaka Mission's Medical College, **Penang International Dental College, Salem; E-mail: [email protected] Objectives: To study the correlation of glycated hemoglobin, total antioxidant status (TAS) and the level of TGF-â 1 in Type II Diabetic patients with renal disease. Methods: The study was conducted on sixty patients with Type-II Diabetes Mellitus, who attended as outpatient department in Madurai Kidney Research Centre, from January-2009 to March-2009. Group I: 26 Diabetic patients without nephropathy (Subgroup IA: 8 patients with good glycemic-control (HbA1C<7%), Subgroup IB: 18 patients with poor glycemic control (HbA1c>7%). Group II: 34 Diabetic patients with nephropathy (Subgroup IIA: 16 patients with good glycemic control (HbA1c<7%) and Subgroup IIB 18 patients with poor glycemic control (HbA1c>7%), Group III: 10 healthy controls. We measured plasma TGF141

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correlation with HbA1c (r=0.4154, p<0.01), creatinine (r=0.2777, p<0.05). Additionally there was a negative correlation between the HbA1c and Results: Our results showed a statistically TAS (r=-0.500, p<0.01). Our results suggest that significant decrease in TAS in all diabetic patients TGF-â1 plays a key role in the development and when compared to controls (p<0.05). TGF-â1 is progression of diabetic-nephropathy and therefore statistically increased in poor glycemic-control apart from glycemic-control and decreasing group of diabetic patients with nephropathy and oxidative-stress, strategies aiming at antagonizing without nephropathy than the control. There was TGF-â1 by the use of a specific-inhibitor of TGFa negative correlation between plasma TGF-â1 â1 may helpful to prevent the development or and the TAS (r=-0.4168, p<0.01), and positive attenuate the progression of diabetic-nephropathy. Abstract No: 3.10

â1, TAS and correlated its level with different biochemical parameters.

Hypothyroidism and Renal Impairment

*K.Shanthi Naidu, **M.R.Chakravarthi, #Bipin Kumar Sethi, *Suryadeep Pratap, *Syed Hyder Ali, *BVLN Murthy and *G.Surekha Departments of * Laboratory Medicine, ** Consultant Nephrologist, # Consultant Endocrinologist, CARE Hospital, Banjara Hills, Hyderabad ­ 500034, Andhra Pradesh

Objective: Knowledge of association between thyroid dysfunction and renal impairment is now an established fact and should be an essential and integral part of evaluation. Methods: The study was conducted retrospectively in hundred samples of renal dysfunction. A control group of both sexes 30 to 70 years considered as normal reference for serum creatinine and TSH. The study group included inpatients (34) and out-patient (66). Assays were processed on Beckman Coulter Synchron Cx9 and Access Results: The clinical presentation of renal impairment was exhaustively studied for an etiological factor which identified hypothyroidism. 80% had elevated creatinine implying renal impairment, of which 75% were

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identified with an abnormal TSH. This small study reveals that severe hypothyroidism may masquerade as renal impairment. Mildly elevated creatinine showed TSH values well above 300 mIU/ml. The identification and treatment soon reduced the renal impairment in terms of lowered serum creatinine and urine protein creatinine ratio along with improvement in the thyroid status. Conclusion: TSH should be considered when evaluating patients of renal impairment of unknown etiology. The cause of renal failure in hypothyroidism may be explained by (i) decreased renal plasma flow due to a hypodynamic state and in severe cases can be (ii) secondary to rhabdomyolysis. Acknowledgements: Nephrology, Endocrinology, Cardiology and Intensive Care Units.

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Abstract No: 3.11

Oxidative Stress and Antioxidant Therapy in Nephrotic Syndrome

Jyoti Dwivedi 1 and Dr.Purnima Dey Sarkar 2, 1 Deptt. of Biochemistry, S.S. Medical College Rewa (M.P.) 486001 India. 2 Deptt. of Biochemistry, N.S.C.B. Medical College Jabalpur (M.P.) India Results: In the present study observed there were significant decreased level of serum total antioxidant capacity, copper, zinc, plasma vitamin C and increased serum level of malondialdehyde, homocysteine, in group 3 when compared group1. However significant reduction in malondialdehyde, homocysteine and significant improvement in serum total antioxidant capacity, copper, zinc & plasma vitamin C were observed Objective: The aim of the present study was to after antioxidant, minerals & B-complex therapy estimate the serum total antioxidant capacity, with routine prescription in group 2 when malondialdehyde, homocysteine, copper, zinc, compared to group 1. plasma vitamin C during remission and Conclusion: In the present study oxidative stress complication. is higher in secondary nephrotic syndrome than Methods: The present study was conducted on 3 nephrotic syndrome. The study further reports groups, group1 comprised of 50 pretreated beneficial effects of antioxidants, minerals and Bnephrotic syndrome patients, group 2 comprised complex vitamins on oxidative stress in nephrotic of 50 post treated nephrotic syndrome patients and syndrome patients may prolong need for treatment group 3 comprised of 41 secondary nephrotic of nephrotic syndrome and secondary nephrotic syndrome patients. Serum total antioxidant syndrome patients. capacity, malondialdehyde, homocysteine, copper, zinc, plasma vitamin C were analyzed. Background: The free radicals have a negative influence on renal tissue in nephrotic syndrome. Nephrotic syndrome is a consequence of an imbalance oxidant/antioxidant status. Hyperhomocyst(e)inemia induce oxidative modification in nephrotic syndrome. Zinc and copper deficiency in nephrotic syndrome related to increase urinary zinc and copper losses. Abstract No: 3.12

Serum Cystatin C levels in Renal Transplant Recipients

Krishnamurthy N 1, Usha Anand 2, Anand C V 2, Aruna V 2, and Venu G 3 1 Department of Biotechnology, Sathyabama University, Chennai, India. 2 Department of Biochemistry and 3 Department of Nephrology, PSG Institute of Medical Sciences and Research, Coimbatore, India. e mail: [email protected] Objective: Serum creatinine (SCr) is a traditional parameter which is widely used for the assessment of renal function. Serum cystatin C (SCys) is an emerging parameter which has been proposed as an early indicator renal dysfunction. This study was carried out to compare the performance characteristics of these two parameters in renal transplant recipients (RTR). Methods: SCr and SCys were estimated using standard laboratory techniques in 30 RTR. Glomerular filtration rate (GFR) was calculated

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from SCr using MDRD formula and from SCys using the Le Bricon formula. Results: Values are expressed as mean ± S.D. The SCr level was 1.257 ± 0.316 mg/dL which is indicative of normal renal function. In contrast, the SCys level was 1.401 ± 0.256 mg/L which is higher than the reference range (0.52 ­ 1.1 mg/ L). The calculated GFR based on MDRD was 65.46 ± 16.37 ml/min/1.73m2 while the GFR based on Le Bricon was 61.59 ± 11.12 ml/min/1.73m2. Abstract No: 3.13

The calculated GFR values are also indicative of mild renal failure. Conclusion: Creatinine is removed not only by glomerular filtration but also by tubular secretion. Cystatin C on the other hand is removed exclusively by glomerular filtration. Levels of SCys therefore rise with even mild decreases in GFR. To conclude SCys is a more sensitive and reliable marker of mildly impaired renal function in renal transplant recipients when compared with SCr.

Serum Zinc and Serum Copper Level in Renal Diseases

Payasvi Baweja*, B.K Agrawal *, and V.K Sharma** * Department of Biochemistry, ** Department of Medicine, Gandhi Medical College, Bhopal, E-mail:[email protected] Objective: Human body contains about 3.07% metals, of which Zinc and Copper are known to be essential factors for growth and development of living things. Its values are found to undergo alterations in various renal diseases. Hypozincemia and Hypocuperemia is found in Nephrotic Syndrome. This study was undertaken to estimate values of Serum Zinc and Copper in renal diseases. Methods: The present study included 54 renal patients and 15 healthy controls matched in terms of Serum Zinc and Copper level by colorimetric method, Akita Abe, Yiamashita, S., (1989), crest biosystems. Results: The Serum Zinc level was Abstract No: 3.14 highly significantly decreased (P<0.001) in Nephrotic Syndrome and Glomerulonephritis patients, while in UTI patients it is significantly decreased (P<0.01) as compared to control group. Serum Copper level was highly significantly decreased (P<0.001) in Nephrotic Syndrome, while in Glomerulonephritis and UTI patients there is no significant difference (P>0.05) as compared to controls. Conclusion: It can be concluded that serial Serum Zinc values are required as an index of prognostic value in renal diseases and estimation of Serum Copper is important diagnostic tool in cases of Nephrotic Syndrome.

Renal Function Screening in Neonatal Asphyxia

Prachi Paliwal,* Dr.B.K.Agrawal,** Dr.Rashmi Dwivedi,***and Dr.Paliwal Manoj**** Gandhi medical college Bhopal (M.P), Mrs. Prachi Paliwal; Email- [email protected] Objective - Birth asphyxia occurs when a baby does not receive enough oxygen before, during or just after birth. Target organs of perinatal asphyxia

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are the kidneys, lungs, brain, liver, gut and bone marrow. The most frequent abnormalities involve Kidneys (50%), CNS (28%), lungs (23%) and so

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on. High blood urea and serum creatinine values are associated with renal failure. We performed this study to determine the incidence of renal failure in birth asphyxia and to correlate the severity of renal failure with asphyxia. By ­: Estimating serum creatinine level in neonatal asphyxia, : Estimation of blood urea nitrogen, : Determination of blood urea nitrogen / creatinine ratio on I, III & VII day of life, : And total urine output within 24 hrs. Methods- The study included 50 asphyxiated and 25 healthy neonates. Renal functions were assessed using urinary output and biochemical parameters such as blood urea by Urease GLDH method and serum creatinine by Jaffes kinetic method. Results- Blood urea and serum creatinine were significantly higher in asphyxiated babies compared to control groups (p<0.001). Blood Urea nitrogen / Creatinine ratio was significantly higher Abstract No: 3.15

in asphyxiated babies compared to control groups (p<0.001) on day 3 but insignificant on day 1. There was no significant difference in the urine output in the control and the study group. Majority of asphyxiated neonates had non-oliguric renal failure. Conclusion- We conclude that renal failure is a significant problem in asphyxiated neonates. A high level of Urea & Creatinine on III rd day of study is associated with renal failure in asphyxia. Despite tremendous advancements in medicine, treatment of ARF has remained essentially supportive. Birth asphyxia is still common, more so in developing countries where obstetric and new born resuscitation facilities are not universally available yet. Combination of dehydration, shock, sepsis and nephrotoxic drugs is not an uncommon situation in NICU.These lead to high incidences of neonatal renal failure. They are often reversible if identified and managed in time.

Exploring the Role of Lipids Isolated from Human Renal Stone Matrix in Urolithiasis

Priyadarshini1, Shrawan Kumar Singh2 and Chanderdeep Tandon1 1 Biotechnology & Bioinformatics, Jaypee University of Information Technology, Waknaghat, H.P., India, 2Post Graduate Institute of Medical Education and Research (PGIMER), Chandigarh, India Background: Organic matrix of calcium oxalate stones consists of protein, lipid, glycosaminoglycans etc. It is suggested that presence of these biomolecules in stone organic matrix is related with the formation of renal calculi. There are many proteins which are reported to exhibit inhibitory or stimulatory activity against kidney stone formation, for example Tamm-horsfall protein, osteopontin etc. However, only few studies have been done on the activity of lipid biomolecules on kidney stone formation. Methods: Surgically removed calcium oxalate stones were first confirmed by FTIR analysis, washed with 0.15M NaCl and then pulverized. Powdered calcium oxalate was extracted with ice cold chloroform: methanol: 0.05M Tris HCl pH7.4 (2:1:1); using sonication at 4ÚC. Upper and lower phases were removed separately and pooled as aqueous and organic substances after centrifugation. Organic phase was extracted with ethanol: ether (3:1) and then centrifuged. Complexed lipids present in pellet were dissolved in DMSO by sonication and were used to check Objective: Therefore, lipid biomolecule(s) were their activity on calcium oxalate (CaOx) crystal chosen to evaluate their activity on the process of nucleation and growth. kidney stone formation.

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Results: The lower volumes of complexed lipids showed inhibitory activity whereas stimulatory activity was observed with their higher volumes when it comes to the nucleation of CaOx crystals. Abstract No: 3.16

However, complexed lipids present in the organic matrix of CaOx kidney stones exhibited inhibitory activity against the CaOx crystal growth assay system.

Prevalence and Characterization of Renal Tubular Acidosis in Patients with Myalgias, Osteoporosis and Mild Arthritis.

Rachel Jacob Department of Biochemistry, NIMS, Hyderabad. Background: Overt metabolic acidosis is undoubtedly associated with metabolic bone disease, but the prevalence of metabolic acidosis in mild osteoporosis and arthritis is uncertain. Chronic metabolic acidosis may also manifest with musculoskeletal symptoms of varying degrees. Objective: It was purposed to identify the prevalence of renal tubular acidosis in patients suffering with chronic myalgias, mild osteoporosis and mild arthritis. Methods: Review of hospital data 2001-2008, revealed 52 patients with complaints of myalgia, muscle weakness, joint pains and/or arthritis. Ammonium chloride load test (0.1g/kg body weight) was done to establish renal acidification defect. Screening for immune-mediated disorders was done. Blood and urine chemistries, arterial blood gas analysis were done to identify electrolyte imbalance. Osteoporosis was assessed by nuclear scan. Abstract No: 3.17 Results: Thirty-four (65%) cases were identified to have an acidification defect(M-14, F-20), 31 had distal tubular acidosis (dRTA) and 3 of proximal RTA. Forty percent of the women were in the menopausal and post-menopausal phase. Neuromuscular weakness associated with hypokalemia was evident in 76% while osteoporosis and bone disease was confirmed in 74%. Metabolic acidosis was seen in 88% with low serum bicarbonate and normal to mild acidosis was noted in 6 cases. One case each of Sjogrens, Vitamin-D resistant rickets, inflammatory arthritis, renal calculus and renal osteodystrophy were identified. Conclusion: RTA can be a major cause of myalgias and joint pains with or without weakness. Women around the menopausal period need to be evaluated for RTA. Normal blood pH or mild acidosis warrants further investigation to avoid missing incomplete dRTA.

Renal Calculi Analysis

K.Shanthi Naidu, Suryadeep Pratap, Zaheed Khan, Syed Hyder Ali, S.M.Ismail and A.Phani Kumar, Department of Laboratory Medicine, CARE Hospital, Banjara Hills, Hyderabad ­ 500034, Andhra Pradesh Objective: Renal calculi analysis is a necessity as the identification of constituent indicates the "type of stone", which is a guide to treatment and

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prevention of recurrence. Renal stones can cause urinary tract infection, damage the kidney and cause obstruction.

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Methods: During a period of six years 197 renal calculi were analyzed. The incidence was seen from 18 to 74 years, predominantly males. The physical features of a stone is important in relevance to its size, weight, colour, texture which have been variable. Biochemical colour reactions of calcium, phosphorus, oxalic acid, magnesium, cysteine, uric acid, carbonate, xanthine were done on the powdered stone material.

that of phosphorus in combination with oxalate, magnesium, xanthine and cysteine. Only a single constituent was seen as a xanthine stone.

Conclusion: In all these patients a complete examination of renal and metabolic status with special reference to aminoacidurias, parathyroid hormones disturbances which may lead to altered calcium phosphorus metabolism are essential to be known, as appropriate treatment and preventive Results: Calcium was seen in nearly 80% of the measures rely on the etiological factor of the stone. stones, its combination being as high as 79.4% with phosphorus 27%, carbonate 15.7%, urate Acknowledgement: Departments of Nephrology 28.4%, magnesium 44.2%, cysteine 6.8% and & Urology xanthine 12.6%. The second largest variety was Abstract No: 3.18

Antioxidant Activity of Creatinine and Uric Acid and Related Efficacy of Antioxidant to Inhibit Glycation

Rohit Suresh*, Yogita S Phapale* and Dr. Z.G Badade** *P.G.Student, **Professor & Head, Department of Biochemistry, MGM Medical College, Kamothe, Navi-Mumbai, Pin-410 209; [email protected]

Background: Non enzymatic glycation, the reaction of glucose and other reducing sugars with protein, reversibly produces amadori products and over a long period irreversibly advanced glycation end product (AGE) are formed. These AGE products contribute to the pathogenesis of diabetic complication. Recently in vitro AGE formation from glucose and albumin were found to be due in part to the oxidants. Objective: The aim of our study is to investigate the antioxidant property of creatinine and uric acid and the inhibition potency of the antioxidant on the formation of AGE products in vitro. In vitro glycation was studied with bovine albumin as the model protein.

Advanced glycation products were quantified by measuring the protein carbonyl content by Levin et al method. Uric acid and creatinine were used as potent inhibition of glycation mixture. Using the same range of concentration used for inhibition of AGE products, uric acid and creatinine were assessed for antioxidant property by reducing power assay. Both antioxidant activity and inhibition of glycation in vitro show a positive correlation. Results: In our study we found that uric acid and creatinine decreased the amount of AGE products in vitro.:

Conclusion: These results underline the importance of uric acid and creatinine as Methods: Albumin was glycated in glucose/ therapeutic agents for the prevention of diabetic fructose mixture by thermal glycation method. and age related complications.

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Abstract No: 3.19

Carbamylated Haemoglobin: A Diagnostic Marker for Chronic Renal Failure.

S.Subramaniam, ShyamaSubramaniam, M.K. Mani*, Partha Karmakar*, K.Ezhil arasan, K.Dhananjayan and M.Gopinath Department of Biochemistry, Apollo Hospitals, Chennai ­ 600 006. * Department of Nephorology, Apollo Hospitals, Chennai ­ 600 006. Background: Carbamylated Haemoglobin is formed by the reaction of haemoglobin with cynate : a product of in-vivo urea dissociation. It is found in high level in patients with renal failure and may be useful in their clinical evaluation. Creatinine, Hb, GlycoHb and Carbamylated Hb. Carbamylated Hb levels were determined in blood samples by HPLC Method using Bio-Rad: D10 GlycoHb Analyser.

Results: Two peaks were seen in control and Acute Objective: To find whether carbamylated Renal Failure patients, whereas only one peak was Haemoglobin can be used as a marker for seen in Chronic Renal Failure patients. If we use the cut off of carrbamylated Hb level 60 u/g Hb, diagnosis of chronic renal failure. then the sensitivity becomes 88.5%, specificity Method: Blood samples were collected from 28 (100%) and Positive predictive Value (100%). normal renal function (Control), 26 Acute renal Failure patients (ARF), 27 Chronic Renal Failure Conclusion: D10 Glycosylated Haemoglobin patients(CRF) with Diabetic nephropathy and 29 analysis graph having a single peak for Non Diabetic nephropathy Chronic Renal Failure Carbamylated Haemoglobin can be used as a Patients. Samples were used for analysis of Urea, reliable marker in identification of CRF. Abstract No: 3.20

Evaluation of Low Dose Doxycycline on Renal Function in Diabetes with Nephropathy

S. Seth, P. Talapatra, H.K. Aggarwal and M. Seth Department of Biochemistry, Medicine & Pharmacy , Pt.B.D.Sharma PGIMS, Rohtak, E-mail ID : [email protected] The study was conducted in forty clinically proven adult cases of Diabetic Nephropathy (Proteinuria >300mg/24 hrs) attending Kidney and Dialysis Clinic at PGIMS, Rohtak. All patients were on stable dose of ACE inhibitors and / or ARB for two months prior to study. Forty patients were divided in two groups. Group-I ­ Patients on insulin / oral hypoglycemic drugs + ACE inhibitors and /or ARB alone. Group-II ­ Patients on insulin / oral hypoglycemic drugs + ACE inhibitors and /or ARB alongwith Doxycyclin (100 mg/day)

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for three months. There was a significant decrease in Proteinuria (urine proteins were estimated by Biuret method) in both the groups when compared to the basal levels. The decrease in group-II was highly significant but the levels returned to the basal levels on stoppage of the drug. There was no alteration in renal biochemical parameters like blood urea, serum creatinine, serum uric acid, serum calcium and serum phosphate. The probable mechanism of action of Doxycyclin in improving the renal function will be discussed.

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Abstract No: 3.21

The Effects of Kidney Function and Transplantation on Plasma Electrolytes, Albumin and Cardiovascular Risk

Thompson, M.J.W.1 and Ray, U.1,2 University of Tasmania, Hobart, Australia1; Royal Hobart Hospital Hobart, Tasmania, Australia2.

Background: Chronic kidney disease (CKD) is Results: Plasma phosphate, magnesium and an increasingly common disease that greatly potassium increased in direct proportion to degree increases risk of cardiovascular disease (CVD), of kidney impairment (all p<0.0001), whereas particularly coronary artery disease. Risk of CVD albumin and calcium decreased (p<0.0001). is directly proportional to degree of kidney Kidney transplantation resulted in an immediate impairment and greatest in end stage kidney reduction in phosphate, magnesium (both disease (ESKD). As CKD progresses, the effect of traditional risk factors such hyperlipidemia on cardiovascular mortality decreases and the effects of novel risk factors such as vascular ossification and hypoalbuminemia increases. Accordingly, clinical trials using traditional interventions such as statins have thus far failed to show a benefit. Optimal treatment for ESKD is kidney p<0.0001) and potassium (p= 0.014). These reductions were maintained long term (all p<0.0001). There was no immediate change in calcium (p=0.34) or albumin (p=0.42), however over a longer time period both increased significantly over pre-transplantation levels (p<0.0001).

transplantation, which is known to improve Conclusion: Kidney impairment resulted in survival over maintenance dialysis. disordered calcium, magnesium and phosphate Aim: To examine the effect of kidney metabolism, which predisposes to vascular transplantation on plasma electrolytes & albumin ossification and increases CVD risk. Kidney to see whether this contributes to the survival impairment was associated with raised potassium, advantage of kidney transplant recipients over pro-arrhythmogenic contributing to mortality. those on maintenance dialysis. Methods: We conducted a retrospective cohort study of 48 patients (M = 29, F = 19; Age = 2484) receiving a kidney transplant at the Royal Kidney transplantation produced an immediate, significant improvement in magnesium, phosphate and potassium levels, which was maintained long term. Significant improvements in plasma

Hobart Hospital within the last 10 years. electrolyte homeostasis and albumin Biochemical profiles immediately pre- and post- concentrations may contribute to the survival transplantation and over the longer term were advantage conferred by kidney transplantation over maintenance dialysis. analysed.

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Abstract No: 3.22

Tacrolimus in Renal Transplant Recipients

Dr. Pradeep Naik and Mallikarjuna.M Department of Clinical Biochemistry, Global hospitals, Hyderabad; Email id: [email protected] Objective: AUC-guided dosing of tacrolimus prevents progressive systemic overexposure in renal transplant recipients. Tacrolimus has a narrow therapeutic drug and bioavailability is known to vary considerably between renal transplant recipients. Most of the transplant centers still rely on measurement of trough levels, but there are conflicting reports on the correlation between tacrolimus trough levels and systemic exposure, as measured by the area-under-theconcentration-over-time curve (AUC(0-12h)). high concentration in 14 patients[ 99 % ] the squared correlation with the AUC (0-12h) (r 2= 0.94).the concentration of four hours post-dose blood is proportional to the increase of the drug dosage .

Results: Compared with trough level monitoring only, this approach reduced the 95%-prediction interval by 50%. The Bayesian approach proved to be feasible in clinical practice, and provided accurate information about systemic tacrolimus exposure in individual patients. In the AUC-guided Methods: We studied and analyzed in 15 renal dosing cohort the apparent clearance of tacrolimus transplant recipients - based pharmacokinetic decreased gradually over time, which was not model with ELISA use of TMB as a substrate to reflected in corresponding trough levels. This estimation of tacrolimus systemic exposure.. The simple, flexible method provides the opportunity main objective was to study intrapatient variability to predict immunosuppression, and should help in the drug concentrations in blood versus different minimize tacrolimus-related toxicity, such as doses. Bayesian forecasting with a two-point nephrotoxicity and post-transplant diabetes sampling strategy, a trough level, and a second mellitus. sample obtained four hours post-dose significantly

Abstract No: 3.23

Study of Microalbuminuria in Hypertension

Sharma, Ashutosh. (1); Lal, A.K. (1); Mehrotra, V. (1) and Verma, S.K. (2) Department of (1) Biochemistry and (2) Medicine, Himalayan Institute of Medical Sciences, Dehradun, INDIA. Email: [email protected] Background: `Save the Kidney' was theme for 2004 given by WHO with the assurance that every hypertensive patient be screened for nephropathy. Microalbuminuria is associated with glomerular damage and endothelial dysfunction which leads to development of nephropathy. Nephropathy, if

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left untreated can lead to serious complication such as kidney failure because albumin molecules are relatively small; it is often among the first proteins to enter the urine after the glomeruli damage. So, even minor kidney dysfunction should be detected with proper diagnosis of microalbuminuria.

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Objective: In our study the effect of hypertension on the incidence and frequency of microalbuminuria has been evaluated and correlated with other renal biochemical parameters. Methods: 60 patients of hypertension of both sexes between 27 to 76 years of age were studied and 20 controls were also included Blood tests (sugar, urea, creatinine and total cholesterol) and urine analysis (protein, sugar and microalbumin was done.

Results: Urinary microalbumin was significantly raised in cases of hypertension. A positive correlation was found between the age and urinary microalbumin levels and serum cholesterol and urinary microalbumin. Similarly creatinine and BUN was raised although within normal range in cases of hypertension with raised urinary microalbumin. It is apparent that the earliest sign of nephropathy in hypertensive patients is microalbuminuria. It is conclude that Nephropathy is a common complication of hypertension and rise in the levels of urinary microalbumin is an early indicator for the development of nephropathy.

Abstract No: 3.24

The Pattern of Nephrotic Syndrome in Malabar region, Kerala

K Sreevidya and George Abraham A.W. H. Special College, Calicut and Malabar Institute of Medical Sciences, Calicut. Email : [email protected] Objective: Nephrotic syndrome or nephrosis is a group of symptoms of kidney malfunction with typical clinical features associated with plasma protein changes such as hypoalbunemia, hyperalbuminuria, hyperlipidemia, hyperlipiduria and edema. There is a geographical variation in the prevalence and pattern of this disease. This disease is mainly seen in children and young adults. Earlier studies conducted revealed a high prevalence of this disease in children in India Subcontinent and South East Asia. Protein electrophoresis is a good screening procedure to evaluate the major serum proteins. This method is extensively used as a diagnostic and prognostic tool for facilitating diagnosis of various diseases, especially nephritic syndrome.many diseases. In this study the serum protein electrophoresis pattern is evaluated using Agarose Gel electrophoresis (Ready to use kit Hydragel Protein kit Supplied by M/S Sebia, France). Methods: A total of 20 serum samples of patients with nephrotic syndrome and serum samples of 20 healthy age and sex matched controls were included for the study. Serum total protein (Biuret method), serum albumin (BCP Dye-binding method) and serum cholesterol (Enzymatic HOD ­ PAP method) were also estimated. Urine samples of all the subjects were collected and analysed. Results: There was a significant increase in serum á ­ 2 globulin fractions and decrease in serum albumin (P d" 0.01) in all patients when compared with that of the normal controls. Decrease in alpha 1 globulin and increase in â globulin fraction is seen in the patients especially in the juvenile group. Decrease in serum albumin is predominant in male patients compared to the females. An increase in cholesterol and â globulin fractions was also seen in patients, predominantly in the juvenile group. Conclusion: The patients of this region presented with lipoid pattern of nephritic syndrome. Contrary

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to the finding of other part of the country, we found more of the disease in adults and old age group (> 50 years). The etiology can be secondary nature, contributed by progressing nephropathy resulted from other diseases. Among our patients the most Abstract No: 3.25

interesting case was that of a lady with 80 years of age presenting with a pattern of á ­ 2 â bridge on the electrophoretogram with a cholesterol concentration of 680 mg/dL, TG ­ 485 mg/dL and a medium albuminuria (80 mg/dL).

Adiponectin Levels in Chronic Kidney Disease and its Correlation with CRP, Cystatin-C and eGFR

Manjunatha G.J and Vijay S. Bhat Manipal Hospital, Old Airport road, Bangalore-560017 Background: CKD is a unique condition in that exceedingly high incidence of insulin resistance, which in turn associated with enhanced cardiovascular events, is paradoxically associated with elevated adiponectin. Objective: Adiponectin, a 30kD collagen like protein exclusively produced by adipocytes, possessing anti inflammatory properties, a marker of oxidative stress was taken up in this study which consisted of patients in different stages of renal disease. A correlative study of adiponectin with markers of inflammation (CRP) and renal function (Cystatin ­C, eGFR & ACR) was done. Results: There was no correlation between CRP and adiponectin levels (p =0.267), There was an inverse relationship between eGFR and adiponectin (ìg/ml log transformed) (r = -0.364, P < 0.001) and a direct relationship with ACR and log transformed adiponectin(r = 0.42, p=0.006). Cystatin­C was significantly higher in renal transplant patients than in normal, independently of serum creatinine & creatinine clearance. Higher adiponectin levels were found with worsening renal function and increasing Proteinuria.

Conclusion: It was observed that in renal transplant patients and those with CKD, the level Methods: Three categories of subjects were taken of adiponectin was nearly double that of baseline. for the study, first group consisting of controls (15 It might be that an increased level of adiponectin members), second group of CKD in various stages is directly involved in impairment of kidney (24 members) and third group who have function, due to their molecular weight. Follow undergone renal transplant (12members). Plasma up data would be invaluable in testing whether our adiponectin was estimated by ELISA method. HSfindings are due to casual relationship, as the CRP and Cystatin­C were Immunonephelometric number of subjects taken in the study was minimal. method. ACR was calculated by estimating urinary It is not known whether this increase in adiponectin albumin & creatinine, eGFR was calculated by using the MDRD equation. Statistical analysis was reflects impaired adiponectin clearance by kidney done by using Kruskal ­ Wallis one way analysis or whether it is a compensatory mechanism aimed at counteracting increased CAD risk factors. of variance and by ANOVA.

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Abstract No: 3.26

Effect of Smoking on Chronic Renal Failure in Primary Glomerular Nephropathies

Prof.(Dr) Uday Kumar* Dr.A.Sharan** and Dr Snigdha*** * Prof & Head,**Additional Professor, Biochemistry,Igims,Patna..** Junior Resident, UCMS & GTBH, Delhi. Background: Smoking is known risk factor for renal damage in diabetic patients. To assess its effect in primary glomerular nephropathy(PGN),we used data from a case;control study designed to assess several environmental risk factors. [ 65%, OR=0.9(95% confidence interval 0.61.4)],but was significantly higher among cases with CRF(75%) than those without [55%,OR=2.4(1.24.5)]. Dose effect relationships were observed with both the daily and cumulative dose;this relationship was stronger in the comparision of cases with CRF and those without CRF than in the comparison of cases with CRF with controls:OR=1.9 versus 1.3(</=20 cigrattes per day)and OR=5.2 versus 3.0(> 20 cigarettes per day): OR=1.9 versus 1.4( </= 15 pac per years) and OR=3.9 versus 2.0( > 15 packs per year). Interactions between age,hypertension and smoking were observed in the risk of CRF: smoking was significantly related to CRF among cases more than 40 years of age and hypertensive.

Methods: The study included 148 biopsy proven glomerular nephropathies of which were 40 membranous nephropathy,58 IgA nephropathy, and 50 nephritic syndrome with either minimal change nephropathy or focal segmental hylinosis.It includes 121 matched hospital controls,with diseases unrelated to smoking. Smoking history was taken from all the subjects. Chronic renal failure(CRF) ,defined by serum creatinine >150 micro mol/L. was present in 37 cases( 28 men & 9 women). Logistic regression Conclusion: This study supports the hypothesis was used to estimate odds ratios(ORS) adjusted that smoking is related to glomerular nephropathy severity,particularly older persons of age group for age and social status. more than 40 and/or hypertensive. Results: Percentage of ever-smokers did not differ glomerular nephropathies cases(60%) and controls Abstract No: 3.27

The Effects of Kidney Function and Transplantation on Plasma Electrolytes, Albumin and Cardiovascular Risk

Thompson, M.J.W.1 and Ray, U.1,2 University of Tasmania, Hobart, Australia1; Royal Hobart Hospital Hobart, Tasmania, Australia2. Background: Chronic kidney disease (CKD) is an increasingly common disease that greatly increases risk of cardiovascular disease (CVD), particularly coronary artery disease. Risk of CVD is directly proportional to degree of kidney impairment and greatest in end stage kidney disease (ESKD). As CKD progresses, the effect of traditional risk factors such hyperlipidemia on cardiovascular mortality decreases and the effects of novel risk factors such as vascular ossification and hypoalbuminemia increases. Accordingly, clinical trials using traditional interventions such as statins have thus far failed to show a benefit. Optimal treatment for ESKD is kidney

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transplantation, which is known to improve survival over maintenance dialysis. Objective: To examine the effect of kidney transplantation on plasma electrolytes & albumin to see whether this contributes to the survival advantage of kidney transplant recipients over those on maintenance dialysis. Methods: We conducted a retrospective cohort study of 48 patients (M = 29, F = 19; Age = 2484) receiving a kidney transplant at the Royal Hobart Hospital within the last 10 years. Biochemical profiles immediately pre- and posttransplantation and over the longer term were analysed. Results: Plasma phosphate, magnesium and potassium increased in direct proportion to degree of kidney impairment (all p<0.0001), whereas albumin and calcium decreased (p<0.0001). Kidney transplantation resulted in an immediate reduction in phosphate, magnesium (both p<0.0001) and potassium (p= 0.014). These reductions were maintained long term (all p<0.0001). There was no immediate change in calcium (p=0.34) or albumin (p=0.42), however over a longer time period both increased significantly over pre-transplantation levels (p<0.0001).

References:Foley RN, Parfrey PS, Harnett JD, et al. Hypoalbuminemia, cardiac morbidity, and mortality in end-stage renal disease. J Am Soc Nephrol 1996; 7: 728-736. Foley RN, Parfrey PS, Sarnak MJ. Clinical epidemiology of cardiovascular disease in chronic renal disease. Am J Kideny Dis 1998; 32 (Suppl 3): S112-119. Hayden MR, Tyagi SC, Kolb L, Sowers JR, Khanna R: Vascular ossification-calcification in metabolic syndrome, type 2 diabetes mellitus, chronic kidney disease, and calciphylaxis- calcific uremic arteriolopathy: The emerging role of sodium thiosulfate. Cardiovasc Diabetol 4:4, 2005 Locatelli F, Marcelli D, Conte F et al. Cardiovascular disease in chronic renal failure: the challenge continures. Registro Lombardo Dialisi e Trapianto. Nephrol Dial Transplant 2000; 15 (Suppl 5): 69-80. Ojo AO, Hanson JA, Wolfe RA, Leichtman AB, Agodoa LY, Port FK. Long-term survival in renal transplant recipients with graft function. Kidney Int 2000; 57: 307­313.

Renal Data System. USRDS 2003 annual data report: atlas of end-stage renal disease in the United States. Bethesda: National Institutes of Health, National Institute of Diabetes and Conclusion: Kidney impairment resulted in Digestive and Kidney Diseases, 2003. disordered calcium, magnesium and phosphate metabolism, which predisposes to vascular Stenvinkel P, Carrero JJ, Axelsson J, Lindholm B, ossification and increases CVD risk. Kidney Heimburger O, Massy Z. Emerging biomarkers for impairment was associated with raised potassium, evaluating cardiovascular risk in the chronic pro-arrhythmogenic contributing to mortality. kidney disease patient: how do new pieces fit into Kidney transplantation produced an immediate, the uremic puzzle? Clin J Am Soc Nephrol 2008; significant improvement in magnesium, phosphate in press. and potassium levels, which was maintained long term. Significant improvements in plasma Wolfe RA, Ashby VB, Milford EL et al. electrolyte homeostasis and albumin Comparison of mortality in all patients on dialysis, concentrations may contribute to the survival patients on dialysis awaiting transplantation, and advantage conferred by kidney transplantation recipients of a first cadaveric transplant. N Engl J Med 1999; 341: 1725­1730. over maintenance dialysis

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Abstract No.3.28

Study of Urolithiasis in Gwalior Region

*A.K. Bhargava, **Neelima Singh, **Sanjeev Singh, **Pradeep Sharma, **Dharmveer Sharma, ***Sunil Agarwal and ****Sunita Sharma *Dept. of Biochemistry, Jhalawar Medical College, Jhalawar; **Dept. of Biochemistry and ***Surgery, G.R.Medical College, Gwalior; ****Dept. of Biochemistry, Madhav Institute of Science and Technology, Gwalior Background: Study of urolithiasis is important in understanding the pathogenesis and medical management of calculus disease. Quantitative analysis of 50 urinary stones (30 kidney, 9 bladder, 11 ureter) collected from different hospitals of Gwalior region was carried out. Objective: To study morphological characteristics of urinary stones. Oxalic acid, uric acid, calcium, and inorganic phosphate were also estimated. Results: The incidence of urinary stones was found as kidney > bladder >ureter The incidence of kidney stones was higher in age of 30-40 than ureter and bladder. Oxalic acid was found as a Abstract no: 4.1 major component in all the stones analysed. Calcium content of stones was kidney > ureter > bladder. Uric acid was seen in stones ureter > bladder > kidney. Inorganic phosphorus content of stone was kidney > bladder > ureter. Hyperoxaluria, hyperuricemia, hyperphosphaturia, hypercalciuria were found in some cases. The high incidence of calcium and oxalate urinary stones in this region may be due to high consumption of vegetables rich in calcium and oxalate. Other factors which may be responsible for urolithiasis are water intake, climatic conditions, dehydration, metabolic abnormalities, urinary tract infections and heredity.

Cardiovascular Risk Markers in Hypothyroidism

Aparna R.Bitla*, E. Jayanti*, Alok Sachan**, G. Shiva Krishna*, M.M.Suchitra*, P.V.L.N.Srinivasa Rao* and KVN Raju* *Department of Biochemistry, **Department of Endocrinology, Sri Venkateswara Institute of Medical Sciences, Tirupati, A.P. Email ID: [email protected] Objectives:cardiovascular disease. Overt and sub clinical hypothyroidism have adverse effect on serum lipid profile that may predispose to development of Atherosclerotic disease. However, the association of the novel risk markers especially Lp(a) and homocysteine in this group Hypothyroidism is a common metabolic disorder and found to be a predictor of of patients is not well known. Hence, the present study was taken up to evaluate the traditional risk marker i.e lipid profile along with the novel markers in patients with hypothyroidism Methods: Twenty five patients with hypothyroidism diagnosed on the basis of TSH measurements were included along with twenty five age and sex matched healthy controls. Lipid profile and Lp(a) were quantitatively measured on Beckmann CX9 fully Automated Analyzer using

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commercial kits. LDL-C was calculated using Friedewald formula. Plasma Homocysteine was measured using a commercial kit by Enzyme linked immunosorbent assay.

Homocysteine (p<0.05) were observed in the study group when compared to the control group

Conclusion: The results of the present study confirm the presence of dyslipidemia in Results: A significant increase in TGL, total hypothyroid patients. This along with the novel risk factors like Lp(a) and Homocysteine which cholesterol, LDL and VLDL (p<0.001) along with were found to be elevated expose these patients to a significant decrease in HDL (p<0.05) were increased cardiovascular risk pointing to the need observed in hypothyroid cases when compared to for institution of appropriate measures early to controls. Among the novel risk marker, a prevent progression significant increase in the levels of Lp(a) and

Abstract No: 4.2

Hyperhomocysteinemia, Circulating Folic acid and Cobalamine Levels in Coronary Syndrome

Yadav AS1, Bhagwat VR2, Mane AY3 and Venugopal NT4 1Dept of Biochemistry, Maharashtra Institute of Medical Sciences & Research, Medical College, Latur (Maharashtra), 2. Dept of Biochemistry, SBH Govt Medical College, DHULE (Maharashtra), 3. Dept of Biochemistry, RD Gardi Medical College, Ujjain (MP), 4. Dept of Biochemistry, DY Patil Medical College, Kolhapur (Maharashtra), Email: [email protected] , [email protected] Introduction: Geographical and nutritional factors are recently shown to have significant role to play in cardiovascular diseases. The exact relationship between nutritional and geographic factors in cardiovascular diseases is not very clear. This study examines relationship of hyperhomocysteinemia with circulating levels of folic acid and cobalamine in patients with cardiovascular disease from rural area. Methods: Blood total homocysteine levels were measured along with folic acid and cobalamine levels in acute coronary and stable angina patients from rural areas along with normal healthy controls from same area. Plasma total homocysteine was measured by HPLC with fluorescence detection. Folic acid and cobalamine

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were measured by chemiluminescence immunoassay. Results: Mean total homocysteine levels were significantly higher by almost three times than the controls. Folic acid and cobalamine were 3-4 times higher in the patients compared to the controls. There was poor correlation between total homocysteine and the vitamin levels in the patients. Conclusion: There is higher prevalence of hyperhomocysteinemia in rural Indian patients. It appears that there is strong association of genetic factors in development of ischemic heart disease in Indian patients. Blood homocysteine is very important biomarker of cardiovascular diseases which must be evaluated with the other risk factors.

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Abstract No: 4.3

Insulin Resistance and Lipid Profile in Pregnancy Induced Hypertension

Asmathulla.S, Rajagovindan.D and Jakanattane Department of Biochemistry, Sri Manakula Vinayagar Medical College & Hospital. Objectives: To study fasting insulin levels and lipid profile in Pregnancy Induced Hypertension (PIH) cases and to compare them with normal pregnant controls. To calculate insulin resistance in PIH and to compare them with healthy pregnant controls. Results: Fasting insulin and HOMA-IR are significantly increased in PIH cases compared to controls. Among PIH cases, Preeclampsia showed significant increase in Insulin and HOMA-IR levels compared to controls and gestational hypertension. Total cholesterol, LDL cholesterol and triacylglycerol are elevated significantly in PIH. The preeclamptic cases are associated with significant rise in total cholesterol, LDL cholesterol, VLDL cholesterol, and triacylglycerol as compared to controls but these changes are not significant in gestational hypertension.

Methods: Twenty eight PIH cases and ten normal healthy pregnant controls are recruited for this study. Serum Insulin levels are measured by ELISA method. HOMA-IR is calculated by (fasting insulin x fasting glucose)/22.5. Total cholesterol, triglycerides, HDL-cholesterol and glucose are measured using enzymatic kits using Conclusion: The data from the present study autoanalyzer. VLDL-cholesterol is calculated by documents Hyperinsulinemia, Insulin Resistance Friedwald's formulae. and Dyslipidemia in PIH patients Abstract No: 4.4

Hypothyroidism Risk factor for Coronary Heart Disease

B.K.Agrawal*, Bhawna Bhimte*, and V.K.Sharma** *Department of Medical Biochemistry, Gandhi Medical College, Bhopal, **Department of Medicine, Gandhi Medical College, Bhopal; Email: [email protected] Objective: Hypothyroidism has been associated with abnormal lipid metabolism, causing dyslipidemia that has been associated with an increased risk of CHD. Level of increased TSH values may be associated with extent of lipid abnormalities. time of detection & second after 6 weeks of therapy: TSH: Competitive Solid Phase Enzyme Immunoassay, Cholesterol: Enzymatic CHODPOD method, HDL: PTA precipitation method, TG: Friedwald's enzymatic method, LDL: Friedwald's formula, Apolipoprotein B: Ag-Ab agglutination method, Lipoprotein a: Ag-Ab Methods :Present study was done in 100 agglutination method. hypothyroid patients & 3 groups have been divided on the basis of TSH level Group I: TSH Results: In Group I: Statistically significant 6-20uIU/dl, Group II: TSH 21-40uIU/dl, Group correlation was found between TC (P<0.05), HDL III: TSH above 40uIU/dl. Following parameters (P<0.001) & TG (P<0.001) whereas LDL values of same patient were analyzed twice first at the were insignificant similarly Apo B were

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moderately significant (P<0.01) & Lip a level were insignificant. Group II: TC, HDL, TG & LDL values were insignificant, whereas Apo B & Lip a were moderately (P<0.01) & highly significant (P<0.001) respectively. Group III: Only the values of TC were found to be significant (P<0.05), whereas HDL, LDL, TG Apo B & Lip a levels were insignificant. Abstract No: 4.5

Conclusion: Above results suggests that hyperlipidemia is associated with extent of hypothyroidism. Not only higher TSH level, but even less raise in TSH level is enough to cause dyslipidemia, increasing the Cardiac risk.

Diabetes Accelerates Age-Related Lipid Profile Disturbances

Zafar M.E.* Nehal M.** Teeto S.A. * and Prasad K.R. * *Bio-Chemistry Department Katihar Medical College, Katihar, Bihar ­ 854105, India, **Ageing Research Laboratory, University Department of Zoology, L. N. Mithila University, Darbhanga ­ 846 004 Objectives:Cardiovascularcomplications (CVC) are predominant problems both in ageing and diabetes. The present study focuses on the effect of human ageing and diabetes , with special reference to serum level lipid profile during cardiovascular complications in two age groups namely, 45 ± 5 years and 65± 5years independent of obesity, hypertension and nephropathy. Methods: The parameters include; estimation of serum total cholesterol, serum triglycerides , HDLC, LDL-C and VLDL-C . Results:Results show that the change in the levels of lipid parameters follow a near similar trend in Abstract No: 4.6 the two non-diabetic age groups with cardiovascular complications. Except the HDL-C decline, total cholesterol, LDL-C, VLDL and triglycerides increase. In diabetes, however, the said lipid parameters are not so similarly affected in the stated age subjects. One interesting observation is the near similar levels of lipid parameters found in diabetic 45 ± 5 yr and nondiabetic 65 ± 5 yr age groups with Cardiovascular complications. Conclusion:It is, therefore, suggested that diabetes accelerates age related disturbances in the lipid profile.

Biochemical Study of Emerging Risk Markers in Coronary Heart Disease

H V Singh,1 S Bhandari,2 N Singh,3 S Singh,3 ..and A Raizada 2

Department of Pathology, Hindu Rao Hospital, Delhi-110007, 2Department of Biochemistry & Cardiology, Escorts Heart Institute& Research Centre, New Delhi-110025, 3Department of Biochemistry, G.R.Medical College, Gwalior-47400 Introduction: To provide the best treatment for coronary heart disease (CHD) patients, one must go beyond LDL cholesterol to detect factors contributing to CHD risk or existing CHD. These

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emerging risk factors appear to identify individuals at increased CHD risk and the assessment of these can modulate clinical judgment when making therapeutic decisions. This study focus on five

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markers lipoprotein (a) (Lp(a)), high sensitive c- or reduced significantly (p<0.001) and it is reactive protein (hs-CRP), Total antioxidant status confirmed by the elevated lipid peroxidation product MDA in both groups. (TAS), homocysteine and Fibrinogen. Methods: Study was carried out by taking 396 subjects, out of which 127 control cases were selected by screening with lipid profile, treadmill test (TMT) and fasting blood sugar, to ensure that they are not suffering from hyperlipidemias, dyslipidemia or diabetes. 269 angiographically proven CAD cases were selected for the present work. Lp(a), hs-CRP, TAS was estimated using Hitachi 912 analyser. The levels of Homocysteine were estimated by EIA method using Bio-Rad kit in Bio-Rad CODA analyzer. The levels of fibrinogen were estimated by Clauss method in Sysmax Autoanalyzer. Results: Recent markers Lp (a), hs-CRP, homocysteine and fibrinogen levels were significantly increased in CAD cases. TAS was significantly decreased in the CAD group. The homocysteine level was significantly increased (p<0.001) in the present study. Oxidative indices like total antioxidant capacity was found decrease Abstract No: 4.7 Conclusions: The statistical analysis of the study shows that these markers are more closely associated with atherosclerosis. Recent cardiac markers i.e. homocysteine and Lp (a) showed no significant correlation in our study with other parameters therefore they may be considered as independent markers. Lp (a) is closely associated with LDL and low values of HDL-C so it could also be considered as independent marker for CAD assessment. The inflammatory markers hs-CRP and fibrinogen emerged as a marker of choice to diagnose atherosclerosis. Many previous studies also showed that persistent hyperglycemia can cause oxidative stress followed by release of inflammatory markers and then formation of atheroma in vessels. The screening with the emerging risk markers is helpful in risk assessment of coronary Heart disease. This additional diagnostic information can assist in the prevention or early diagnosis and treatment of atherosclerotic cardiovascular disease.

Anti-oxidant Potential (AOP) of LDL in Post Menopausal Women Suffering from Type 2 Diabetes Mellitus.

Nivedita Singh, Neelima Singh, Sanjeev Singh and Vishal Bhargava Department of Biochemistry, G.R. Medical College, Gwalior Objectives: Menopause occurs as the age advances in life of women (40-50 years). Post menopausal phase is associated with absence of estrogen known to be cardio protective and potent antioxidant hormone. A strong association of absence of estrogen and cardiovascular complication in post menopausal diabetic female has been reported. years)and 50 post menopausal diabetic female subjects.

Results: It is concluded from study that in these subjects besides hyperglycemia, hypertryglyceridemia and hyperchole-sterolemia also exists. LDL and VLDL were increased (P<0.001) with decrease of HDL cholesterol (P<0.001). Sensitivity for oxidation of LDL in Methods: The present study has been carried out these subjects were found increased and therefore in 50 age matched healthy female (40-50 antioxidant potential (AOP) of LDL was decreased

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Abstract No: 4.8

Increased Levels of Inflammatory Markers, Sialic Acid and Cardiovascular Risk Factors in Type 2 Diabetes Mellitus.

Dr S.Rajesh1, Dr S.Sethupathy2 and Dr S.M.Rajendran3 Division of Biochemstry, Chettinad Hospital and Research Institute, 2. Division of Biochemistry, Raja Muthiah Medical College, Annamalai University, 3. Division of Medicine, Raja Muthiah Medical College, Annamalai University Objective:Type II DM patients generally carry a number of risk factors for CVD. Microalbuminuria, Serum sialic acid (TSA) and elevated levels of inflammatory markers, particularly hs C-reactive protein have been shown to be cardiovascular risk factors. There are no satisfactory data on circulating concentrations of inflammatory cytokines and their potential relationship with traditional and nontraditional cardiovascular risk factors in a diabetic population. The aim of the present study was to examine the relation of serum cytokines ,microalbumiuria and serum sialic acid concentrations, to cardiovascular risk factors in type2 diabetic patients. necrosis factor , Interleukin-6, C-reactive protein, Lipid profile, Apo-A1 and Apo-B were estimated. Results: Statistical analysis was done using Systat ver. 12.0 software. RESULTS ­Highly significant association was found between Microalbuminuria and Crp(0.001),Sialic acid (0.001).TNF and Apo B (p=0.02),Total cholesterol(p=.03), IL-6 and ApoB (P=0.01),Apo-A(P=0.08), Sialic acid and ApoB(P=0.017),Totalcholesterol(p=0.00), Apo A(p=0.000) , TGL(p=.01).CRP and Apo-B (p=.02), Apo-A(p=.001).

Conclusion: The present study provides evidence of a significant increase in inflammatory markers, Methods: we enrolled 80 type 2 diabetic patients, sialic acid and cardiovascular risk factors in type without any major systemic illness. 2 diabetics with increased urinary albumin Microalbuminuria, Serum sialic acid, Tumor excretion. Abstract No: 4.9

Influence of Atherosclerosis and Exercise on Arterial Lipoprotein Lipase Activity in Pigs

Shakti Aggarwal, Theodore W. Zderic, Perminder Gulani and Marc T. Hamilton, University of Missouri-Columbia, USA. Objective: Lipoprotein lipase (LPL) has repeatedly been identified as one of the major susceptibility genes for human coronary artery disease. However, the functional enzyme activity has heretofore never been quantified in arteries with respect to either vascular disease or exercise training. We have overcome prior experimental limitations by utilizing a porcine model of atherosclerosis, where the endothelium from large

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arterial samples could be obtained in conjunction with modifications in the LPL assay to improve accuracy. The primary purpose was to determine the effects of exercise training on arterial LPL activity in a porcine model of atherosclerosis. Methods: Heparin releasable LPL (HR-LPL) activity was measured in approximately 5.2 cm2 of the thoracic aorta, skeletal muscle and adipose

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tissue. In keeping with the purpose, a pig strain with familial hypercholesterolemia (FH) was run trained for ~5 months and compared to controls fed an identical diet (N=13 run trained and N=12 untrained). As a basis of comparison to generally healthy pigs, LPL was measured in another strain of pigs with substantially lower plasma cholesterol (Yucutan).

19±3 mU/mg total protein; p<0.05). This two-fold training effect for artery HR-LPL was evident in both males and females. In contrast to this, neither skeletal muscle, nor adipose HR-LPL activity was altered by the exercise training. Paradoxically, arterial HR-LPL activity was 13.7 fold greater in FH pigs (21.3±4.6 mU/mg), compared to the Yucatan pigs (1.6±0.2 mU/mg).

Results: There was about twice as much arterial Conclusion:: Exercise run training increased HR-LPL activity in FH pigs after the exercise arterial HR-LPL activity in a porcine model of training compared to sedentary controls (40±4 vs. lipoprotein-induced atherosclerosis.

Abstract No: 4.10

Study of Apolipoprotein A-1, B and Lipid Profile in Premenopausal and Postmenopausal Women

Dr. Swapnali* and Dr. D.S. Jayaprakash Murthy. Department of Biochemistry, J.J.M. Medical College, Davangere-577004, Karnataka. Email: [email protected] or [email protected]

Results: TC, TG, very low density lipoprotein (VLDL), low density lipoprotein (LDL), Apo B, TC/HDL and Apo B/Apo A-I were increased, whereas HDL and Apo A-I were decreased in postmenopausal women when compared to premenopausal women and were statistically Objective: So this study was conducted to as- significant (p < 0.001). sess the relationship between serum Conclusion: Apo A-I and HDL cholesterol are apolipoprotein (Apo) A-I, Apo B and lipid pro- antiatherogenic whereas Apo B and LDL cholesfile in healthy premenopausal and postmeno- terol are atherogenic. The cholesterol content of pausal women. HDL and LDL as an indicator for risk of CAD Methods: About 123 healthy subjects were studied (62 premenopausal and 61 postmenopausal women). Total cholesterol (TC), triglycerides (TG) and high density lipoprotein (HDL) were estimated by enzymatic method and Apo A-I and Apo B were estimated by immunoturbidimetric method by semiautoanalyzer. may be misleading because the cholesterol content varies with a variety of physiological and pathological conditions but not their protein content. Hence estimation of Apo A-I and Apo B, the protein part of HDL and LDL respectively serves as a more reliable tool in predicting the risk of CAD in postmenopausal women

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Background: Menopause is an estrogen deficient state. Natural menopause confers a threefold increase in coronary artery disease (CAD) risk. Currently postmenopausal women account for more than 30% of the female population at risk for CAD in India.

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Abstract No: 4.11

Significance of Plasma Total Homocysteine Levels in Type II Diabetes Mellitus Ppatients

Anbazhagan M and Ben S. Ashok Dept. of Biochemistry, Sri Ramachandra University, Porur, Chennai. Background: Diabetes mellitus is a common disease with many complications. CVD is the major cause of death in Type II diabetes mellitus. In recent years several studies have demonstrated that elevated homocysteine is a risk factor for atherosclerosis. Elevated homocysteine levels increase the risk of every complication associated with type II diabetes. This study was aimed to determine the relationship between blood glucose, Total cholesterol and fasting total homocysteine (tHcy) levels in patients with Type II diabetes with no renal complications. levels were measured by GOD-POD method and Total cholesterol were measured by CHOD using Hitachi 902 analyzer in 110 diabetics patients and 60 non diabetics. Results: Plasma tHcy levels were found to be significantly high in diabetics than non diabetics (p< 0.01). Total cholesterol levels were also found to be significantly high in diabetics than non diabetics (p<0.01).

Conclusion: As hyperhomocysteinemia also reflect endothelial injury, the present observation Methods: Plasma tHcy was measured by support the hypothesis that hyperchemiluminescence assay using siemens Advia homocysteinemia appears as an risk factor for centaur. Fasting and post prandial blood glucose CVD in type II diabetes patients. Abstract No: 4.12

Apolipoprotein A1 Gene Polymorphism (G-75A and C+83T) in Patients of Myocardial Infarction- a Pilot Study in North Indian Population.

Rajni Dawar1 *, Anil Gurtoo2 and Ritu Singh1 1 Department of Biochemistry, Lady Hardinge Medical College & Associated S.S.K Hospital. New Delhi-110001. India, 2Department of Medicine, Lady Hardinge Medical College & Associated S.S.K Hospital. New Delhi 110001. India. Objective: To study whether genetic polymorphism in ApoA1 gene (G-75A and C+83T) could be playing a role in occurrence of Myocardial Infarction. Methods: Study group consisted of 100 subjects (50 diagnosed MI patients, 50 healthy subjects). S. Apolipoprotein A1 and S. Apolipoprotein B were estimated by immunoturbidometric assay on SYNCHRON CX9 (BECKMAN). Extracted

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DNA from blood sample was amplified by PCR, which was digested overnight with MspI restriction enzyme. Digested products obtained were run on 8% Polyacrylamide gel and Restriction fragment length polymorphism was studied by gel documentation system. Results: S. ApoA1 levels were significantly higher in control group compared to study group (100.80 ±7.06mg/dl and 72.56±9.86mg/dl respectively;

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p<0.0001) and S. Apo B levels were significantly higher in study group compared to control group (97.45 ± 9.04mg/dl and 72.12 ± 11.32mg/dl respectively; p<0.0001). The frequency of the G allele at -75bp site was higher in the study group (79%) compared to controls (58%).Genotypic distribution in study group was [GG (58%),GA(42%)] compared to controls [GG(16%),GA(84%)] with p<0.0001. The frequency of the T allele at +83bp site was higher in the study group(56%) compared to controls(32%).Genotypic distribution in study Abstract No: 4.13

group [CC(24%),CT(40%),TT(36%)] was different as compared to controls [CC(36%),CT64%)] with p<0.0001. Conclusion: In Apolipoprotein A1 gene, G at 75bp upstream from start of transcription and T at +83bp site in the first intron may be susceptibility alleles for myocardial infarction in Indian Population. However, ours being a pilot study, more studies with larger sample size are needed to confirm genotypic risk associated with G and ­ (T) allele.

Role of Pro-inflammatory Cytokines, ApoB and Small Dense LDL in Women with Coronary Artery Disease

Sunita1, S.B Sharma1, D.Puri1, R.L.Tripathi 1 and S.Dwivedi2 Department of Biochemistry1 and Medicine2, University college of Medical Sciences, Delhi, India. Objective: Inflammation plays important role in initiation and progression of atherosclerosis. Therefore assessment of Pro- inflammatory markers such as hs-CRP,IL-6,ApoB and small dense LDL help in the prediction of risk and can aid in treatment plan in female patients of CAD who present with atypical symptoms so that mortality associated with CAD can be decreased. highly significant (p<0.001) in MI & UA as compared to controls.LDL-C was also significantly increased (P<0.05) in MI as compared to UA.TG and HDL-C were also increased but not at the significant level(p>0.05).ApoB ,hs-CRP & IL-6 were highly significant(p<0.001) in UA & MI as compared to controls.Small dense was highly significant(p<0.001) in CAD as compared to controls.96.7% of UA & 83.3% of MI showed Methods: Present study comprises 90 female presence of small dense LDL. subjects of age 40-65 years which included 30 healthy controls, CAD patients into 30 cases of Conclusions: Levels of TC, LDL-C were high in unstable angina (UA) & 30 cases of myocardial patients of UA & MI. ApoB, hs-CRP & IL-6 were infarction (MI).Fasting blood samples analyzed high in patients of CAD as compared to controls. for lipid profile, ApoB using standard methods. These high levels reflect severity of CAD & thus hs -CRP and IL-6 were analysed by ELISA. Small help in risk stratification .Small dense LDL was dense LDL was calculated by ratio of LDL-C/ highly increased in UA as compared to MI which ApoB. suggest that small dense LDL are precursor of atherogenesis. Results: Total cholesterol (TC) & LDL-C were

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Abstract No: 4.14

Dyslipidemia and Anti-oxidant Nutrient in Pre-eclampsia

Dr.M.G.Dhabe*, Dr. (Mrs.)S.C.Choudhary** and Dr.M.R.Mogarekar* *SRTR medical College,Ambajogai,Dist Beed, Maharashtra.**B.J.Medical College,Pune. Objective: Preeclampsia is the main cause of maternal and perinatal mortality and contributes 8% of maternal deaths in India. In women with preeclampsia there is dyslipidemia and decreased antioxidant activity. The present study was undertaken to evaluate the lipid (cholesterol& TG) levels and serum ascorbic acid level in preeclampsia. Methods: For the present study, we have selected 40 normotensive pregnant women of age group 20-35 years comprising Group I and 40 preeclamptic women of same age group comprising Group II. Serum samples were analysed for serum total cholesterol, triglycerides and serum ascorbic acid levels. Abstract No: 4.15 Results: A significant increase in serum total cholesterol and serum triglyceride levels were found in Group II as compared to Group I while serum ascorbic acid levels were decreased significantly in Group II as compared to Group I. Conclusion: In preeclampsia there is dyslipidemia. The hypertriglyceridemia may lead to increased endothelial triglyceride accumulation that in turn may result in endothelial dysfunction probably by promoting oxidative stress in arterial wall. The antoxidant nutrient like ascorbic acid may be utilised to greater extent to counteract free radical mediated cell disturbances resulting in reduction in antioxidant nutrient level in serum.

Post Coronary Stenting Decrease in Atherogenic Risk Factors

Kavitha S1, Sridhar M G1, Satheesh S2 and Balachander J2 Department of 1Biochemistry and 2Cardiology, JIPMER, Puducherry Objective: To study the effect of coronary stenting estimated before and three days after coronary on atherogenic risk factors in patients with stent surgery using paired-t-test. coronary heart disease (CAD). Results: A significant reduction was observed in Methods: A total of 24 acute coronary syndrome total cholesterol (157.6±32 vs. 140.4 ±29.9, (ACS) patients who underwent coronary stenting p=0.01), triglyceride (195.5±85 vs. 147±59, were enrolled in the study. The study group p=0.001), total cholesterol/HDL (3.9±0.7 vs. included patients with acute ST-elevation 3.6±0.7, p=0.05) and triglyceride/HDL (5.1±2.5 myocardial infarction (STEMI) and non­ST vs. 3.9±1.7, p=0.004) levels after coronary elevation ACS, (NSTEACS). Lipid profile stenting. There was no significant difference in parameters and atherogenic indices like total HDL­Cholesterol, LDL- Cholesterol and HDL/ cholesterol/HDL, triglyceride/HDL were LDL after three days of coronary stenting.

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Conclusion: The present study shows that a stenting. However factors causing this decrease significant decrease in atherogenic risk factors is need further evaluation. found in CAD patients following coronary Abstract No: 4.16

Serum Uric Acid in Acute Myocardial Infarction

Gargi Sen and Virupaksha H. S. Department of Biochemistry. J.S.S. Medical College and Hospital, S.S.Nagar.Mysore-15, Karnataka. serum uric acid was high in M.I patients. The association of uric acid with Killip class at day 0,3,7 was analyzed and the classification of uric acid was carried out as normal < 7 mg/dl and abnormal >7 mg/dl. It was observed that at day 0, 3 and 7 there was significant association between Objective: To estimate serum uric acid in patients Killip class and uric acid levels. No statistically with M I and correlate it with Killip class, FBS, significant association was observed between uric HbA1C, Blood Pressure and obesity. acid levels and age, sex, hypertension, diabetes Methods: 30 patients with myocardial infarction mellitus and obesity individually. and 30 controls were studied. Serum uric acid was Conclusion: Uric acid levels were higher in measured on day 0,3,7 of M I. FBS, HbA1C, patients of myocardial infarction and correlated Blood Pressure and Body Mass Index was noted. with Killip class independent of other cardiac Results: Statistically significant higher levels of biomarkers. Combination of Killip class and serum uric acid was found in M.I patients on day 0,3,7 uric acid levels after MI, could act as a good as compared to controls ,thereby indicating that prognostic indicator in these patients. Abstract No: 4.17 Background: High uric acid is considered a negative prognostic marker in heart failure. Close association exists between uric acid and Killip classification of acute myocardial infarction patients.

Apolipoprotein B, Total Cholesterol, HDL-C, Non HDL-C, LDL-C and Triglycerides as Risk Factors of Atherosclerotic Vascular Disease

Jitty George, Dr.Andrews M.A, Dr.Cibu Mathew, Dr.Geetha Damodaran K;, Dr.Gilsa E 1.Sixth-semester MBBSstudent, 2. Professor of Medicine, 3. Assistant Professor of Cardiology, 4. Associate Professor of Biochemistry, 5. Senior Lecturer of Biochemistry; Email ID:[email protected] Objective: To find out relative significance of serum levels of ApolipoproteinB versus Total Cholesterol, HDL cholesterol, Non HDL cholesterol, LDL cholesterol and Triglycerides in atherosclerotic vascular disease. Methods: Design: Observational case control study. Setting: Medical College Chest Hospital, Thrissur. We measured fasting serum Apolipoprotein B and fasting lipid profile. Inclusion criteria: Patients of any age with atherosclerotic vascular disease Controls:Clinically normal persons Exclusion

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criteria: Patients already on lipid lowering drugs Controls: Those with diabetes and hypertension and those on lipid lowering drugs Apolipoprotein B was measured by immunoturbidimetry; total cholesterol by enzymatic method; HDL by phosphotungstic precipitation method; LDL by indirect method; Triglyceride by enzymatic method. Non-HDL was calculated as TC minus HDL-C. Semi auto analyzer was used for all measurements. Fasting blood sample was used for all assys. Data analysis: ATPIII guidelines were used to select cut off values:Total cholesterol>200mg%, HDL- C<40mg%, LDLC>100mg%, Non HDL-C >130mg% ,TG>150mg%.Cut off for apolipoproteinB(105mg%) adopted from a previous study done by Sakurabayashi.

Results: The values of TC(p=0.484),NonHDL­ C(146±102 vs 130?80;p= 0.386), LDL-C(121±96 vs 108 ± 78;p=0.895) and TG(129 ± 106 vs 107 ± 100;p= 0.746) are found statistically insignificant.Only HDLC(5 ± 32 vs 63 ± 26;p=0.025) and apolipoproteinB(116 ± 46vs95 ± 28;p=0.001)are found statistically significant. Conclusion: ApolipoproteinB value of 105mg% (P<0.001) is found to be most significantly associated with atherosclerotic vascular disease.Hence measurement of ApolipoproteinB levels is highly recommendable in the risk assessment of atherosclerotic vascular disease for early recognition and prevention.Moreover, it can be performed on nonfasting blood sample also.

Abstract No: 4.18

Accuracy of Retinal Changes in Predicting Microalbuminuria in Obese and Nonobese Hypertensive Patients

Arul Senghor*, Ebenezer William* and Jothimalar** *SRM medical college Potheri, **SRMC Porur Objective: Hypertension and obesity is becoming a common health problem worldwide. It is linked with vascular inflammation, lipotoxicity and oxidative stress that results in vascular end organ damage. Aim of the study is to measure the inflammatory markers of vascular endothelium, namely hs-CRP and urinary microalbumin in obese and non-obese hypertensives, that predicts the future cardiovascular events. Methods: The study was conducted among 120 subjects who were non-smokers and non-alcoholic of age 40 ­ 60 yrs with hypertension. Patients were divided into three stages of hypertension according to JNC-6. Fasting blood samples were collected to estimate glucose, lipid profile, blood urea nitrogen, creatinine, hs-CRP, TC, DC. Overnight spot urinary sample was collected to measure

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urinary microalbumin. Fundus changes were observed to assess the vascular endothelial damage. Results: The mean of hs-CRP and microalbumin was significantly high in obese than non-obese hypertensive group (p= 0.000). There was a strong correlation between hypertensive retinal changes and microalbuminuria (P < 0.0001 ). Tests of accuracy for retinopathy as a predictor of microalbuminuria showed a sensitivity of 72% and specificity of 81%. Conclusion: hs-CRP and urinary microalbumin are early markers of endothelial dysfunction which can predict cardiovascular risk in hypertensives. Retinal changes have moderate accuracy in predicting microalbuminuria

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Abstract No: 4.19

Circulating Levels of Cell Adhesion Molecules in Hypertension

Kavita K. Shalia, Manoj R. Mashru, Jagdish B. Vasvani, Rajashree A. Mokal. Shilpa M. Mithbawkar and Priti K. Thakur Sir H. N. Medical Research Society, Sir H. N. Hospital and Research Centre, Mumbai. Objective: Hypertension causes complications such as coronary atherosclerosis and thrombosis wherein inflammatory factors play significant role. In the present study inflammatory molecules such as cell adhesion molecules (CAMs); endothelial (E)-selectin, platelet (P)­selectin, intercellular CAM-1 (ICAM-1), vascular CAM-1 (VCAM-1) and platelet endothelial CAM-1 (PECAM-1) were analysed in subjects newly diagnosed with hypertension with no secondary cause against normotensive healthy individuals. Methods: In each group 57 subjects were recruited and soluble (s) levels of CAMs were analysed by ELISA. Medians were compared by Mann Whitney U test and correlation was analysed by Spearman Correlation. Abstract No.4.20 Results: As compared to controls median of sEselectin, sP-selectin and sICAM-1 were significantly elevated by 49.2%, (P=0.001), 54.3% (P=0.001), 18.9% (P=0.012) respectively in hypertensive subjects. Significant negative correlation was observed of sP-selectin (spearman rank correlation coefficient (rs) =-0.345, p=0.027) and sPECAM-1 (rs =-0.446, p=0.003) with age in hypertension group. Conclusion: Hypertension may increase expression of certain CAMs while younger hypertensives in addition are also at increased risk of atherothrombosis.

Serum Paraoxanase Activity and its relationship with Atherogenic Index of Plasma (AIP) in Hyperlipidemic Patients

Kavya R, Phani M.N, Supriya.M, Jeevan K.Shetty and Mungli Prakash Department of Biochemistry, Kasturba Medical College, Manipal -576104; E-mail id: [email protected] Objective: Paraoxonase (PON 1) is a high density lipoprotein associated enzyme, is proposed to have an antiatherogenic effect by preventing LDL oxidation. Atherogenic index of plasma (AIP) is the index used to predict the risk of atherosclerosis. Aim of our present study was to investigate the relationship between AIP and PON 1 activity in newly diagnosed hyper lipidemic patients. hyperlipidemic patients who came for routine check up without any systemic disease and 50 healthy controls. Lipid profile was analyzed by auto analyzer and PON 1 activity were determined spectrophotometrically using substrate paraoxan.AIP was calculated using formula log [TG/HDL-C].

Results: There was significant increase in TC, TG, Methods: The serum lipid profile, PON 1 and AIP LDL-C, AIP (p<0.001) and significant decrease in were estimated in 50 newly diagnosed HDL and PON 1 (p<0.001) in cases compared to

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controls. There was no significant correlation between PON 1 & AIP. Conclusion: According to our study, hyperlipidemic patients has got decrease in PON Abstract No: 4.21

1 activity, increase in AIP without having significant correlation between them. This suggests decrease in PON 1 activity and increase in AIP is the independent risk factors for development of atherosclerosis

Trends in Blood Pressure with Increasing Serum Homocysteine Levels

Kumar. Animesh1, Gupta. P. R2and Ali. Z3 Biochemistry Department, 1Himalayan Institute of Medical Sciences, Dehradun, 2 Cardiology Department and 3 Biochemistry Department, IMS B.H.U. Varanasi. Email: [email protected] Introduction: It has been predicted that by 2020 there would be a 111% increase in cardiovascular deaths in India due to hypertension resulting in cardiovascular diseases (CVD). Mild elevations in serum homocysteine may contribute to elevations in blood pressure which may be one of the important causes for CVD. Controversy remains as to whether the relation between homocysteine and CVD is causal or not? Our study comprises the relationship of homocysteine with blood pressure. Methods: Our study consists of 40 subjects diagnosed as hypertensive for the first time and 20 controls with normal Blood pressure taken randomly from cardiology OPD of our hospital. Most of the patients of both sexes were from Eastern Uttar Pradesh (Varanasi) and from Bihar. Abstract No: 4.22 They were in the range of 25 to 60 years of age. Levels of Homocysteine were measured in serum by ELISA technique and Blood pressure along with clinical history was observed separately for all subjects. Results: It was observed that Homocysteine was significantly elevated in hypertensive patients as compared to controls with p<0.001. It also bears a high degree of correlation with systolic blood pressure. It was also observed that the sex along with age and life style plays a very important role for our study. Conclusion: This study shows that homocysteine is significantly elevated in patients of hypertension and concludes that in every CVD patients homocysteine plays a very important role.

Coronary Heart Diseases and Serum Triglycerides

K Ram achandran, Divya K.V., Ajeena and Sheela Email ID: [email protected] Objectives: There are many etiological factors responsible for the development of coronary heart diseases (CHD) and there are many cardiac markers for the assessment of CHD. The present study was undertaken to assess whether the various

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cardiac markers pertain or indicate the etiology of CHD. Markers studied in this study are Total cholesterol, HDL cholesterol, LDL cholesterol, Triglycerides, CK-MB, AST and Troponin I. Etiological factors for CHD in the present study

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are 1. Long standing Diabetes Mellitus (DM), 2. Hypothyroidism, 3. Nephrotic syndrome, 4. Liver cirrhosis. Other estimations done in this study are blood glucose,serum ALP, blood urea and serum creatinine.

fasting blood glucose estimation, lipid profile, enzyme studies, urea and creatinine. Results: 1. Group 1 and Group 3 - high fasting blood glucose (value more than 180mg%), 2. Group 1 and Group 2 - high total cholesterol, high LDL cholesterol, high triglycerides, high VLDL. 3. Group 3 and Group 4 - normal cholesterol 4. HDL cholesterol was found satisfactory in all the groups 5. AST and ALP upper normal limit in group 2 only. 6. Blood urea and serum creatinine normal in all groups 7. Serum creatinine - normal in all groups.

Methods: Total cholesterol by using cholesterolesterase - Cholesterol oxidase ­ Peroxidase, Triglycerides - GPO-PAP end point assay, HDL cholesterol - precipitation method using phospho tungstate and magnesium acetate reagent, LDL cholesterol - calculated by Friedwald's formula, CK-MB - based on immunoinhibition method using G6PD, AST- by Conclusion: 1. The present study established a coupled enzyme using malate dehydrogenase, close relationship between DM and CHD, 2. Total Troponin I - by card test cholesterol is elevated in these two conditions and Blood glucose - by GOD-POD method, ALP- by is responsible for CHD, 3. HDL cholesterol was PNPP method, Blood urea - by GLDH method, found satisfactory in DM and CHD which is a Serum creatinine - Jaffe's method. 60 patients significant finding compared to other study -that admitted in Co-operative medical college, Kochi is HDL cholesterol may not truly represent coming under the age group 50 to 82 years were atherosclerosis in physically active people, 4. selected. They were grouped into Group 1 - DM Triglycerides and VLDL represent the occurrence + CHD, Group 2 - CHD only, Group 3 - DM of CHD and atherosclerosis, 5. There is no without CHD with 20 patients in each group. involvement of hepatic an renal dysfunction in the Group 4 - Normal (20 people). Blood sample was occurrence of CHD with DM. collected from all persons from all groups for Abstract No: 5.1

Investigation Profile Correlated with Adverse Maternal and Perinatal Outcome in Pregnancy Induced Hypertension (PIH)

Ms.Liggy Andrews, Dr.N.Haridas* and Dr. Smruti Vaishnav** 1.Dept of Biochemistry, ** Dept of Obstetrics & Gynecology. Ms. Liggy Andrews, Department of Biochemistry. C.U.Shah medical college. Surendranagar. Email Id: [email protected] Background: PIH is still a major cause of of biochemical profile of adverse pregnancy Obstetrical and Perinatal morbidity and mortality. outcome in PIH From the stand- point of prevention, PIH has Method: Study was conducted in Biochemistry, remained a constant challenge to the obstetrician. Obstetrics & Gynecology Department at Objective: To evaluate & correlate the significance S.K.Hospital, Karamsad, Gujarat-January 2006 to

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March 2008.Total 328 PIH &100 normal pregnant AST* & APTT* - significantly associated with women were studied. All biochemical tests maternal mortality. While urea*, creatinine*, APTT*, uric acid*, calcium*, LDH*, ALT & AST analyzed on Xl-300. - significant with Perinatal mortality. Uric acid Results:Total no. of deliveries -- 2237. Out of >6.0mg/dl in 9 maternal and 66 perinatal deaths which 328 had PIH (prevalence 14.7%).Out of while LDH >600IU/l in 50 perinatal deaths which mild PIH -- 117(5.2%), severe PIH 136(6.0%), Eclampsia 67(2.9%) and chronic HT Conclusion: Standard antenatal follow up should 8(0.35%). Complications were hematological be carried out for the early detection and (22.5%) -- DIC(3.0%), HELLP syndrome (3.9%) prevention of adverse consequence related to PIH. and thrombocytopenia (15.5%) was the most All biochemical markers should be carried out in common maternal complication associated with each PIH patient to help predict and to deal with mortality and morbidity. 9(2.7%) maternal deaths the adverse pregnancy outcome and and 86(26.2%) Perinatal deaths (SB 56 & 29 complications.*(P<0.000) NND) occurred in this study. Biochemical parameters were analysed. Uric acid*, ALT*, Abstract No: 5.2

Status of Lipid Profile in Patients of Acute Myocardial Infarction

K.Suchitra, Bari Siddiqui M A and U Satyanarayana Dept of Biochemistry, Siddhartha Medical College, Dr NTR University of Health Sciences, Vijayawada, Andhra Pradesh 520008. [email protected] Introduction: Alterations in the lipid profiles have been reported in patients of acute myocardial infarction (AMI). These reports however are controversial- some claim reduction while others report increase in different parameters of lipid profile. The objective of the study was to evaluate the changes that occur in lipid profile in AMI, including the ratios of total cholesterol / HDL cholesterol, and LDL cholesterol / HDL cholesterol. Methods: This study was conducted in 25 patients of AMI admitted at Government General Hospital, Vijayawada. Fasting(14 hr) blood samples were collected at 24 hours, 48 hours and 5 days predischarge, and serum was analyzed for TG, total cholesterol, HDL cholesterol, VLDL and LDL cholesterol were calculated using Friedwald formula.

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Results: There was no significant change in the serum total cholesterol at 24 hours, 48 hours and 5 days pre- discharge. HDL cholesterol marginally fell at 48 hours; VLDL showed slight increase at 48 hours and 5th day pre- discharge. Changes in LDL cholesterol were negligible. Serum TG showed an increasing trend after MI at 48 hours and 5th day pre- discharge. An elevation in the ratios of total cholesterol / HDL cholesterol was observed at 48 hours and 5th day pre- discharge. However, LDL / HDL ratio remained almost the same. Conclusion: It is concluded that although there are variations (24 hours, 48hours and 5th day predischarge) in the lipid profile in the patients of AMI, the ideal period for the estimation of lipid profile appears to be within 24 hours of the acute episode.

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Abstract No: 5.3

A Study of Age Related Changes in the Serum Lipid Levels in Healthy Adults of Mangalore City

*Dr.Malathi.M, and Ms.Sangita Kamath Professor, Dept of Biochemistry, Fr Muller medical college,Mangalore. E mail : [email protected] Background: Coronary artery disease (CAD) is the most common type of heart disease.. Hyperlipidemia is considered as one of the risk factors in the development of coronary artery disease .(CAD) . The National Cholesterol education program (NCEP) has suggested normal reference ranges for lipid profile. However various epidemiological studies done in different parts of India show different cut off levels suggesting that the lipid levels could be influenced by various geographic factors .These studies point out the need to survey the population region wise and to establish normal reference ranges in them. Objectives: To assess the average lipid levels in the healthy adults of Mangalore city and to set up age and sex matched reference ranges in the population. healthy controls of age groups between 21-75 yrs was assayed in the laboratory using autoanalyser by the recommended methods .The results were then categorized age wise under four groups . Stastical analysis was done by student t test and ANOVA (analysis of variance) methods. Results: The overall lipid level was found to be on the higher side of the normal reference level in the study population. In males lipid levels were found to be higher in 36-50 yr age group and in females in the postmenopausal group who were beyond 50 yrsof age. Conclusion: Screening for dyslipidemia in all adults above 20 years and Health education regarding modification of life style and dietary habits would be useful to control hyperlipidemia.

Methods: Total cholesterol, LDL cholesterol,, Acknowledgment: This study was done as ICMR HDL cholesterol and Triglyceride levels of 200 STS project in 2005 Abstract No: 5.4

Systemic Failure of Interferon-á synthesis, a Potent Inhibitor of Platelet Aggregation and a Thrombolytic Agent, and its Role in the Development of Acute Myocardial Infarction.

Mau Bhattacharyya, and Asru K. Sinha* Sinha Institute of Medical Scince and Technology, 288 Kendua Main Road, Baishnabghata, Garia, Kolkata-700084, E mail- [email protected] Objectives: We have reported before that the increase of nitric oxide synthesis in leucocytes in the circulation resulted in the synthesis of interferon-á (IFN-á), a well known cytokine. It was also found that IFN-á was a potent inhibitor of platelet aggregation (minimal inhibitory dose

10nM) similar to that of prostacyclin (PGI2),

currently believed to be the most powerful inhibitor of platelet aggregation. However unlike PGI2, IFNá was a potent thrombolytic agent that dissoluted clotted plasma in vitro and fibrinolysed coronary

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thrombosis in animal model. We investigated the role of IFN-á in the pathophysiology of the development of AIHD. Methods: Fifty patients with acute myocardial infarction (35 male, 15 female, between 50-68 year old who were admitted in ICU, Calcutta Medical College, Calcutta) and 50 age and sex matched normal volunteers participated in the study. IFNá was determined by enzyme linked immunosorbent assay (ELISA). Nitric oxide was assayed by methemoglobin method.

Results: Plasma IFN-á level in normal volunteers was 17.09±7.49 pmol/ml incontrast the IFN-á level in AIHD patient was 1.6±0.053 pmol/ml (p<0.0001). The plasma nitric oxide level in normal was 0.4±0.02 nmol/h decreased to 0 pmol/ h in AIHD patients. After recuperation the impaired IFN-á and NO levels in AIHD were found to return to normal level. Conclusion: Severe reduction of IFN-á synthesis due to the decreased synthesis of NO resulted in the impaired thromboprotection by the cytokine in AIHD. And, as such the decreased plasma IFNá level might be a hither to unrecognized risk factor for AIHD in men.

Abstract No: 5.5

Atherogenic Index in Patients with Established Myocardial Infarction.

Abdul Lateef , Afshan Rafi and Pragna Rao Department of Biochemistry, Kamineni Institute of Medical Sciences, Narketpally,508254.AP. [email protected] Introduction: Atherogenic Index defined as log(TG/HDL-c in mmol/l) has recently been proposed as a marker of atherogenicity.An abnormal ratio of Triglycerides(TG) to HDL indicates an atherogenic lipid proûle and a risk for the development of Myocardial infarction. Objective: To estimate and compare the Atherogenic Index in patients diagnosed with MI with that of controls. Methods: 100 Patients attending cardiology department with symptoms of Acute MI with high Cardiac Troponin T(cTnT) were selected as patient group, and healthy subjects were selected as control group. Serum Total Cholesterol(TC),HDL and TG were measured using COBAS INTEGRA, and cTnT by using COBAS E411 Electrochemiluminescence. Results: Patients were divided into 3 groups depending upon cTnT levels.

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Group1(controlgroup):cTnT<0.010ng/ml Group2:cTnT upto 0.1ng/ml and Group3:cTnT>0.1ng/ml. In Group1:The Mean and SD of Atherogenic Index was 0.22±0.18, TG:135.31±46.21mg/dl, TC:149.85±34.09mg/ dl,HDL: 33.69 ± 6.87 mg/dl. In Group 2: Atherogenic Index was 0.36 ± 0.22, TG: 222 ± 119.73 mg/dl, TC: 204.69 ± 56.70 mg/dl., HDL: 35.69 ± 5.10mg/dl. In Group3: Atherogenic Index was 0.32 ± 0.14, TG: 180.45 ± 56.69mg/dl, TC: 181.96 ± 48.35 mg/dl., HDL: 34.66 ± 5.67mg/dl. Atherogenic Index was significantly increased in patients with microinfarcts (p<0.0001) when compared with normals.Patients with frank infarcts had higher Atherogenic Index when compared to patients with microinfarcts.( p>0.05 ). Conclusion: Other than serum choleterol levels, elevated triglcerides and lower HDL significantly contribute to elevated atherogenecity and increase the risk of MI.

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Abstract No: 5.6

Increased Cardio Metabolic Risk Factors seen in School Cildren

Sumathi, Meera , Vinodhini , and Ebenezer William SRM Medical College Hospital & Research Centre, Potheri. Objective: To study the occurrence of metabolic (p<0.001) in obese children than in controls. The risk factors in obese children compared to healthy levels of total cholesterol, triglyceride and LDL were significantly increased (p<0.001) among the children. obese. HDL level was significantly decreased Methods: School children aged 8 ­ 15 years (n= (p<0.001) in obese than the controls. The insulin 150) from diverse socio economic background level in obese group showed significant positive were studied. Demographic, social and medical correlation with Body mass index, Waist hip ratio details were obtained, anthropometry and blood and total cholesterol, thus indicating insulin as a pressure were measured. Fasting plasma glucose, component of lipid and glucometabolic risk factor. insulin and lipid profile were measured. Conclusion: Obese children showed higher Results: 65% of obese children showed abnormal prevalence and clustering of abnormality compared values for one or more cardio metabolic risk to controls. The final conclusion is obese children factors. (i.e., fasting plasma glucose, lipids and are at an increased risk for the development of insulin). Obese children had higher prevalence cardiovascular events at an earlier age compared and clustering of abnormalities compared with to non obese children. normal children. The levels of fasting blood glucose and insulin were significantly elevated Abstract No: 5.7

Assessing Risk Factors in Acute Coronary Syndrome

Moushumi Lodh*, Ashok Parida* and Alpana Saxena** *Dept of Biochemistry and Dept. of Cardiology, The Mission Hospital, Durgapur; **Dept. of Biochemistry, MAMC & LN Hospital, New Delhi. Objectives: PAPP-A and PlGF are abundantly expressed in ruptured and eroded plaques. PAPPA, a matrix metalloproteinase, stimulates IGF-1 and is said to have pro atherosclerotic action. PlGF, a member of VEGF family, expressed from inflammatory cell types , stimulates pathological angiogenesis. The goal of this study was to determine predictive value of PAPP-A, PlGF in acute coronary syndrome and to look for relationship with markers of myocardial necrosis (CKMB, troponin I), inflammation (hsCRP, leptin). Methods: 100 clinically diagnosed acute coronary syndrome patients with angiographically documented significant coronary artery disease were evaluated within a few hours of reporting to emergency department with chest pain. Comparisons were made with 60 age matched normal healthy controls . Patients of advanced kidney or heart failure , major surgery/trauma within the previous month, known inflammatory diseases, suspected thrombotic disorders were excluded. Serum PAPPA, PlGF, Lp (a), hsCRP and leptin levels were assayed , correlations

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checked with quantitative troponin I and CKMB pregnancy associated plasma protein A levels mass levels. showed significant correlation with hsCRP levels(p<0.05). Results: Mean hsCRP , PAPP-A, CKMB mass and Troponin I levels showed significantly higher Conclusions: Elevated PAPP-A and PlGF levels levels in cases compared to controls. PlGF levels in ACS patients may reflect instability of in patients was higher but the difference was not atherosclerotic plaques. They may be an early statistically significant. In the patient group, serum indicator of ACS . Abstract No: 5.8

Atherogenic Index of Plasma in Ischaemic Heart Disease Patients.

Phani M.N, Supriya .M, Kavya R, Jeevan K Shetty and Mungli Prakash Department of Biochemistry, Kasturba Medical College. Manipal, India-576104. E-mail id: [email protected] Objective: Atherosclerosis is the common cause for ischaemic heart disease (IHD) and the mortality and morbidity related to it. There are various indicators to predict the risk for development of atherosclerosis. The aim of our present study was to measure atherogenic index of plasma (AIP) and TC/HC ratio in IHD patients and to check the relationship between them. Methods: Serum lipid profile, AIP and TC/HC ratio was estimated in 32 IHD patients and 32 healthy controls. Lipid profile was analyzed by auto analyzer and TC/HC was calculated. AIP was calculated using formula log [TG/HDL-C]. Results: There was significant increase in TC, TG, Abstract No: 5.9 LDL-C, AIP, TC/HC (p<0.001) and significant decrease in HDL (p<0.001) in cases compared to controls. Significant positive correlation was observed between AIP and TC/HC. Conclusion: According to our study IHD patients are more prone to the development of atherosclerosis due to high TC/HC ratio and AIP when compared to healthy controls. AIP can predict the size of LDL particle and high LDL levels and size are important factors in development of atherosclerosis. Along with TC/ HC, AIP can also be used in routine laboratory to predict the risk for development of atherosclerosis in IHD patients.

Current Biomarkers for Myocardial Infarction

Rathish R* and Sumathi P Department of Biochemistry, SRM Arts and Science College, Kattankulathur, Kanchipuram ­ 603 203 Tamil Nadu. Objective: Ischemic heart disease is the leading cause of death in developed countries, but third to AIDS and lower respiratory infections in developing countries. Myocardial infarction (MI) is a common presentation of ischemic heart

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disease or coronary artery disease. The current study focused on the lipid profile patterns and cardiac biomarker levels in MI patients and role of lipid profile and biomarkers with the risk of MI. Role and sensitivity of Troponin I over other

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biomarkers. The study was carried out in three in comparison with other groups. MI prone subjects groups: normal subjects, MI prone subjects and were found to have discordant biomarker levels in MI patients. their blood with normal Troponin I, SGOT levels and elevated CK, CKMB levels. Methods::Lipid profile and cardiac biomarkers like Troponin I, CK, CKMB, SGOT were analyzed Conclusion: The present study reveals that MI in all three groups. prone patients can be detected by their lipid profile patterns and discordant biomarker levels. From Results: The lipid profile patterns were found to these results, it could be concluded that Troponin be significantly high in MI patients and MI prone I is specific and sensitive to cardiac damage, can category. MI patients were found to have elevated be used for the detection of MI and CVD risk. Troponin I levels (80 fold) and other biomarkers Abstract No: 5.10

Appearance of Dermcidin, a Heat Shock Protein, as a Risk Factor for Acute Ischemic Heart Disease through the Inhibition of Pancreatic Insulin Synthesis

Rajeshwary Ghosh and Asru K. Sinha* *Sinha Institute of Medical Science and Technology, 288 Kendua Main Road, Baishnabhghata, Garia, Kolkata-700084, E mail- [email protected]

Objective: The occurrence of hyperglycemia is commonly associated with acute myocardial infarction and unstable angina. The association of hyperglycemia is reported to result in the worst prognosis of the functional recovery in this condition. The mechanism in hyperglycemia in AIHD remains obscure. We investigated the role of an inhibitor in the circulation in AIHD for the inhibition of glucose induced pancreatic insulin synthesis. Methods: The synthesis of insulin was determined by in vitro translation of the isolated mRNA from the islets of Langerhans from the mice pancreas exposed to glucose and determined by enzyme linked immunosorbent assay (ELISA). Results: The treatment of the islets of Langerhans with the plasma from AIHD patients but not with

the plasma from normal volunteers in the presence of glucose resulted in the inhibition of insulin synthesis from 0.148µunits of insulin /mg of islets/ h in the control experiment to 0.012 µunits of insulin /mg of islets/h in the presence of AIHD plasma. This inhibitor from the AIHD plasma was isolated by preparative SDS gel electrophoresis. The isolated protein band (14kD, a single polypeptide chain) that was found to inhibit glucose induced insulin synthesis was determined to be Dermcidin isoform 2, an oxidative stress protein by the amino acid sequence analysis. Conclusion: The appearance of Dermcidin in the circulation in AIHD, would superimpose a prothrombotic condition over the existing AIHD through the impairment of insulin which is established to be a potent antithrombotic hormone in recent time.

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Abstract No: 5.11

Uric Acid Levels in Cardiovascular Disease

Mrs. Ranjita V.Gaur, Dr.(Mrs.)Rita M.Shah e mail id :[email protected] / atmiya04yahoo.com Objective: The purpose of this study was to see the relationship of elevated serum uric acid level in cardiovascular disease. (>8.41mg/dl) had significantly higher than females (>6.73mg/dl). Mean SUA.

Conclusion: Our data suggest that increased serum Methods: The study has been carried out in Dhiraj uric acid levels are independently and significantly General Hospital, Pipariya, Baroda with known associated with risks of cardiovascular disease case of cardiovascular disease in 100 patients. mortality. Results: Cardiovascular events occurred in 10% 0f patients. In males Serum uric acid level Abstract No: 5.12

Effect of a Single Dialysis Session on Plasma Lp(a) Levels in Patients on Maintenance Haemodialysis

Seshadri Reddy. V1, M.M. Suchitra1, E. Prabhakar Reddy1, Aparna R.Bitla1, V. Sivakumar2 and P.V.L.N. Srinivasa Rao1 Department of Biochemistry1, Department of nephrology2, Sri Venkateswara Institute of Medical Sciences, Tirupati, A.P, India.E-mail: [email protected] Background: Cardiovascular disease (CVD) is a major cause of mortality in End stage renal disease (ESRD) patients on Maintenance haemodialysis (MHD). Lp (a), is a specialized form of glycoprotein-LDL-cholesterol complex and is an independent risk factor for myocardial infarction. The risk is related to its atherogenic and thrombogenic properties. Lp(a) were estimated on Beckmann CX9 Fully Automated Analyzer using commercial kits. Statistical analysis was performed using SPSS for windows version 11.5.

Results: Results of analysis of variance for repeated measures after correction for hemoconcentration where necessary revealed a decrease in Lp(a) (p=0.022) and triglycerides Objective: The present study was taken up to (p=0.001) levels and no change in cholesterol evaluate changes in Lp(a) and Lipid profile in (p=0.48) levels. patients undergoing hemodialysis session. Conclusion: Maintenance dialysis program is Methods: Twenty seven patients with end stage known to produce Dyslipidemia. Study of Lp(a) renal disease who were on maintenance in dialysis patients is important as this is an hemodialysis were included. Plasma samples were independent risk marker. However there are very collected hourly during a dialysis session with few reports on changes in Lp(a) due to the dialysis polysulfone membrane using bicarbonate session. Our findings will be discussed in dialysate. Plasma cholesterol, triglycerides, and comparison with other reports.

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Abstract No: 5.13

Association of LDL Particle Size with other Cardiovascular Risk Markers in an Industrial Population.

Mohammad Tarik, Lakshmy Ramakrishnan, Ruby Gupta, Dorairaj Prabhakaran , and Kolli Srinath Reddy Department of Cardiac Biochemistry, All India Institute of Medical Science, New Delhi-29. Email: [email protected] Objective:LDL is heterogeneous in terms of size, density, electric charge and chemical composition with some particles of LDL (small dense LDL particle) showing greater atherogenecity compared to other particles. Studies on LDL heterogeneity are not available in resident Indians. Since data is not available in Indians it would be pertinent to look at LDL heterogeneity in a resident Indian population and look at their association with other known biochemical risk factors of CAD like lipids, Apo A, Apo B, Lipoprotein (a) and hsCRP. Methods:183 employees of an industry in Delhi with age range 30 -39 years were screened for their LDL particle size in plasma by 3% polyacrylamide gel electrophoresis. Their lipids, Apo A, Apo B, Lipoprotein (A) and CRP were estimated. The study included 140 males and 43 females. Results: 41 males (29.3%) and 11 females (25.6%) had LDL particle size less than 26.3 nm. Abstract No: 5.14 The group with LDL particle size d" 26.3 nm (pattern B) (n=52) were compared with the group having LDL particle size > 26.3 nm (pattern A) (n=131). Females with particle size d" 26.3 had significantly higher total cholesterol, LDL-c, lipoprotein (a) and hsCRP than females with particle size > 26.3 nm. Among males, Apo B was significantly higher in the d" 26.3 group. Apo B, lipoprotein (a) and hsCRP was significantly higher in the < 26.3 group when males and females were combined. Conclusion:A high prevalence of small dense LDL (particle size < 26.3 nm) was observed in the industrial population studied. There was a significant difference in lipid parameters in the subjects with the A and B phenotype. This confirms that the atherogenic LDL particle size (pattern B) is associated with other cardiovascular risk factors in Indians.

A Novel Antihypertensive Protein Hormone from the Kidney Cortex, and its use for the Diagnosis of Essential Hypertension in Men.

Somashree Chakraborty, Gausal A. Khan, and Sinha A.K* Sinha Institute of Medical science and Technology, 288 Kendua main road, Baishnabghata, Garia, Kolkata-700084, E mail- [email protected] Objective: Essential hypertension, the form of hypertension in men, is a major risk factor for coronary artery disease (CAD). No etiological reasons, no gene or any definable cause for essential hypertension is currently known. And, as such, there is no diagnostic test for the condition which,by itself does not produce any discernible symptom in the victims. We have recently reported the existence of an antihypertensive protein hormone produced in the kidney cortex and named the protein "cortexin".

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Methods: To determine whether of plasma cortexin level in humans will provide a diagnostic test for essential hypertension, blood samples from the persons with established essential hypertension and age and sex matched normotensive subjects were collected. Cortexin level was determined by enzyme-linked immunosorbent (ELISA) assay using anticortexin antibody. NO was determined by the formation of methemoglobin.

cortexin, originally was identified in the goat kidney cortex, cortexin was found to occur in the blood of all animals including men. It was found that plasma cortexin level as determined by ELISA in normotensive persons was 218.94 pmol cortexin/ ml (n=50) that contrasted 0 pmol cortexin/ml in person with established essential hypertension (n=50) (p<0.00001).

Conclusion: These results indicate that the Results: Cortexin was found to normalize elevated determination of plasma cortexin level in men may blood pressure both systolic and diastolic pressure be useful in the diagnosis of essential hypertension through the activation of endothelial nitric oxide in man. synthase (NOS) in animal model. Although Abstract No: 5.15

Ischemia Modified Albumin ­ an Evolving Diagnostic Marker of Myocardial Ischemia

Dr Sudeshna Behera and Prof. P.C.Mohapatra Department of Biochemistry , S.C.B. Medical College, Cuttack, Orissa, INDIA; Email id ­ [email protected] Background: Establishing a diagnosis of Acute Coronary Syndrome (ACS) in clinical setting remains a challenging task. Currently cardiac markers like Myoglobin, Cardiac Troponin T (CTnT) and CK-MB are commonly used in assessing cellular necrosis in patients attending emergency department with symptoms of acute chest pain. However there is no accepted gold standard for detecting myocardial ischemia. So it is difficult for the clinicians to prevent the progression of myocardial ischemia to irreparable myocardial damage. Recently a new marker Ischemia Modified Albumin ( IMA ) has been introduced as a highly sensitive and early detectable marker of myocardial ischemia. Objective: The present study was conducted to evaluate serum IMA concentration in acute coronary syndrome patients and to assess its diagnostic importance in cases of suspected myocardial injury. Methods: The study evaluated IMA in conjuction with ECG and CTnT in thirty five ACS patients

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presenting to the emergency department within 3 hrs of acute chest pain. thirty healthy volunteers served as control. The levels of IMA were determined by Albumin Cobalt Binding (ACB) test and readings were taken at 470 nm by spectrophotometer. Serum MDA and Vitamin E levels were also assessed in all patients. Results: Serum IMA levels were observed to be significantly raised (P<0.05) in acute coronary syndrome patients than controls. Rise in IMA demonstrated an association with MDA and Vitamin E revealing its genesis due to oxidant stress. The high sensitivity of IMA alone as well as in combination with ECG and CTnT makes it a better and an early diagnostic marker of myocardial ischemia. Conclusion: IMA can be measured accurately , reliably and within an acceptable time period to be useful in the evaluation of patients with ACS. It could bring a new dimension to the care and management of patients at an early stage.

Indian Journal of Clinical Biochemistry Supplement, 2009/24

Abstract No: 5.16

Study of Serum Magnesium, HbA1c , Lipid Profile and Microalbuminuria in Diabetic Retinopathy

Swetha N.K, and Virupaksha H.S. Dept of Biochemistry , J S S Medical College Mysore-15,Karnataka; Email id:[email protected] Background: Diabetic retinopathy is a sight threatening complication of diabetes mellitus and is one of the leading causes of acquired blindness. There is a series of risk factors related to the development and progression of diabetic retinopathy such as hypomagnesemia, dyslipidemia, duration of diabetes, poor diabetes control. Objective: 1. To estimate serum magnesium, HbA 1c , total cholesterol, triglyceride and microalbuminuria in type II diabetic patients with and without retinopathy. 2. To compare and see if there is any significant correlation between these parameters in diabetic patients with retinopathy. 3. To compare the levels of these parameters with the healthy volunteers. Methods: The study included 30 diabetic patients with retinopathy, 30 diabetic patients without Abstract No: 5.17 retinopathy and 30 healthy controls. Serum magnesium, HbA1c, total cholesterol, triglyceride and microalbumin in urine were estimated. Results: There was a significant increase in the FBS, HbA1c and microalbumin in urine in diabetic patients with retinopathy and also in diabetic patients without retinopathy, when compared to the control group. There was a significant decrease in the serum magnesium levels in the diabetic groups when compared to the controls. However there was no significant variation in the total cholesterol levels in the study groups. The severity of diabetic retinopathy in the studied groups was influenced by factors such as duration of diabetes mellitus, good glycemic control, hypomagnesemia and microalbuminuria

The Effects of Kidney Function and Transplantation on Biochemical Correlates of Cardiovascular Health

Thompson. M.J.W. 1 and Ray1.U. 1,2 , University of Tasmania, Hobart, Australia1; Royal Hobart Hospital Hobart, Tasmania, Australia2. Background: It is consistently now being appreciated that chronic renal dysfunction alone is an independent risk factor for the development of coronary artery disease, and for more severe coronary heart disease (CHD)[1,2,3,4]. One in seven Australians show signs of chronic kidney disease (CKD). CKD is a potent, independent risk factor for cardiovascular disease (CVD). Risk of CVD is directly proportional to degree of kidney impairment. As well as independently increasing the risk of CVD, kidney dysfunction also adversely impacts upon other biochemical determinants of cardiac health. Kidney impairment increases total cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL), urates & decreases high density lipoprotein (HDL). It has also been suggested that kidney impairment induces a state of insulin resistance in the absence of pre-existing diabetes

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mellitus or other comorbidities know to cause insulin resistance. Insulin resistance results in resulting in impaired glucose metabolism and hyperglycaemia, further worsening CVD risk profile. These biochemical derangements are seen in even the earliest stages on kidney impairment. Thus kidney function has an important role in maintaining cardiac health. Optimal treatment for end-stage kidney failure (ESKF) is kidney transplantation. Previous studies have shown a decrease in CVD risk in patients who underwent kidney transplantation compared those patients who did not. However, CVD risk in patients who receive a transplanted kidney remains elevated above the general population. We propose that improved plasma lipid profile and reduced inflammation may contribute to this decrease in CVD risk. Objective: To examine the relationship between kidney function, lipid profile and urate and see how these are affected by kidney transplantation in non-diabetic patients with ESKF. Methods: We conducted a retrospective analysis of non-diabetic patients (M= 15, F= 7, Age = 1972) who had received a kidney transplant within the last ten years at the Royal Hobart Hospital. Patients with diabetes were excluded on the basis of request notes and clinical history. Only patients who had baseline measurements prior to transplantation and post transplantation measurements were included in the study. Results: HDL decreased with increasing degree of kidney dysfunction (p<0.0001) with even mild degree kidney impairment impacting negatively on HDL (p= 0.006). LDL, TC, TG & urate increased with increasing degree of kidney dysfunction (p<0.0001), despite statins use in >90% of patients. These were seen to increase even in mild stage kidney failure (p<0.0001). Kidney transplantation resulted in a statistically significant increase in HDL (Student's t-test,

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p<0.001) and also decreased TG levels (Student's t-test, p= 0.014) when compared to pretransplantation levels. LDL, TC & urate were also decreased following kidney transplantation, however the effect was not significant. Data were also analysed using Piecewise 3-step regression to examine temporal trends in HDL, LDL, TG, TC and urates in pre- and post-transplant subjects. Conclusion: Kidney impairment results in changes to lipid profile and inflammatory status that negatively impact on cardiovascular disease risk. Changes in lipid profile and inflammatory status are observed even in the earliest stages of kidney failure. Kidney transplantation has a positive effect on inflammatory status & lipid profile, particularly HDL. Reference: 1: Sarnak, MJ, Levey, AS, Schoolwerth, AC, Coresh, J. Kidney disease as a risk factor for development of cardiovascular disease: a statement from the American Heart Association Councils on Kidney in Cardiovascular Disease, High Blood Pressure Research, Clinical Cardiology, and Epidemiology and Prevention. Circulation 2003; 108:2154. 2: Ix, JH, Shlipak, MG, Liu, HH, Schiller, NB. Association between renal insufficiency and inducible ischemia in patients with coronary artery disease: The Heart and Soul Study. J Am Soc Nephrol 2003; 14:3233. 3: Chen, J, Muntner, P, Hamm, LL, Jones, DW. The metabolic syndrome and chronic kidney disease in U.S. adults. Ann Intern Med 2004; 140:167. 4: McCullough, PA, Nowak, RM, Foreback, C, et al. Emergency evaluation of chest pain in patients with advanced kidney disease. Arch Intern Med 2002; 162:2464.

Indian Journal of Clinical Biochemistry Supplement, 2009/24

Abstract No: 5.18

Utility of NT proBNP in a wider range

*K.Shanthi Naidu, *Suryadeep Pratap, *Syed Hyder Ali, *BVLN Murthy, * B.Goud, *B.Raghavendra and *J.Visalasree Departments of Laboratory Medicine, CARE Hospital, Banjara Hills, Hyderabad ­ 500034, Andhra Pradesh Objective: NT ProBNP is a well known biological marker of left ventricular dysfunction. The interpretation of abnormally elevated values threw light on other cardiac manifestations with variable clinical presentation. of cardiac evaluation. Elevation of more than 3 ­ 5 fold increase was observed in patients of pregnancy induced hypertension and eclampsia when compared to a mild increase in normal pregnancy. Pleural effusion evaluation showed elevated levels signifying an underlying cardiac pathology. Patients of renal impairment had elevated levels and needed aggressive cardiac evaluation for left ventricular dysfunction. An increase of natriuretic peptide was also seen in septic patients.

Methods: Proven diagnosis of 25 patients of myocardial infarction, 5 of pulmonary embolism, 20 of normal pregnancy, pregnancy induced hypertension and eclampsia, 10 patients of dialysis, 25 patients of pleural effusion were analysed for NT ProBNP on Elecsys (Roche) Conclusion: NT proBNP widely used for LV 2010/1010. dysfunction when studied in various clinical Results: Patients of myocardial infarction had NT manifestations can throw light on asymptomatic ProBNP in highly elevated and was correlating cardiac conditions calling for a high grade of with Troponin evaluation. Pulmonary embolism cardiac evaluation. diagnosed by imaging and D-Dimer showed a gross increase in absence of left ventricular Acknowledgement: Cardiology, Nephrology, Gyn dysfunction, suggesting a need for higher grade and Obstetrics and Intensive care units Abstract No: 5.19

Normalization by Aspirin of Hypertension for the Reduction of Risk of Coronary Artery Disease in Men

Uttam K. Maji and Asru K. Sinha* Sinha Institute of Medical Science and Technology, 288 kendua Main Road, Baishnabghata, Garia, Kolkata-700084, E mail- [email protected] Backgrfound: Hypertension is known to be a major risk factor for acute ischemic heart disease. However the normalization of the elevated blood pressure using currently available antihypertensive therapy therapeutic regimen resulted in only 8% decrease in the risk of acute ischemic heart disease. We have recently reported that synthesis of cortexin, an antihypertensive protein hormone in the kidney cortex cell. None of antihypertensive drug is capable of stimulating cortexin synthesis. Systemic increase of NO stimulated cortexin synthesis and acetyl salicyclic acid (aspirin) stimulates systemic NO synthesis Objective: we determined the effect of oral aspirin (150mg/75kg) ingestion on the elevated systolic

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and diastolic blood pressures. Cortexin was also found to be a potent inhibitor of platelet aggregation.

mmHg at presentation were found to become normalized (123.33±10.30 mmHg systolic, 82.5±5.5 mmHg diastolic respectively) after 3h of ingestion of aspirin with concomitant increase of Methods: Newly diagnosed patients with cortexin level from 0 nM (before aspirin ingestion) hypertension, (n=50) who never received any to 9.23± 0.042nM (after the ingestion) of the antihypertensive medication at presentation were compound (p<0.0001) in these patients. asked to participate in the study by ingesting enteric coated aspirin (150mg/75kg) only after the Conclusion: Since cortexin is reported to be a participants took a full meal. The blood pressures potent inhibitor of platelet aggregation,and was were measured after 3h of ingesting the drug. found to be a powerful normalizer of hypertension, Cortexin was determined by enzyme linked aspirin might be useful for the reduction of risk factor for CAD both through the normalization of immunosorbent assay (ELISA). the hypertension and inhibition of platelet Results: The both systolic and diastolic pressure aggregation. which were 160.63±13.85 mmHg, 96.63±8.03 Abstract No: 5.20

A Study of Insulin Levels in Pregnancy Induced Hypertension

Vinodhini, Meera Shivshekar and Ebenezer William Department of Biochemistry, SRM Medical College Hospital & Research Centre , SRM University, Potheri, Tamil Nadu Background: Preeclampsia one of the most common complications of pregnancy is a major cause of maternal, fetal and neonatal morbidity and mortality. Many features of insulin resistance syndrome have been associated with this condition. Objective: The aim of this project was to study the levels of insulin and lipoproteins in pregnancy induced hypertension (PIH). Methods: The study included 30 patients with PIH, 20 patients with normotensive pregnancy and 20 age matched controls. Abstract No: 5.21 Results: The mean level of insulin was significantly elevated in cases of PIH than the controls (P< 0.001). The levels of total cholesterol, TGL and LDL were significantly high and HDL was significantly low among patients with PIH than the controls. Conclusion: The result of this study indicates that insulin resistance plays a role in pathogenesis of PIH. Such patients are at an increased risk for future cardiovascular disease. Interventions to reduce insulin resistance will minimize later life cardiovascular complications.

Levels of GHBA1C in Non- diabetic Patients with Cardiovascular Disease

Tarun Kumar Sharma**, H. Parchwani*, G.G. Kaushik* and Vijay Shankar** *Department of Biochemistry, J.L.N. Medical College, Ajmer (Rajesthan), **Pt. B.D. Sharma, P.G.I.M.S., Rohtak (Haryana) Objective:High levels of GHbA1C traditionally have been associated with cardiovascular disease

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(CVD) and diabetic patients. The available data suggests that an association exists between levels

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of GHbA1C and CVD, even in patients without diabetes. observed 4.32 + 0.34% and 5.80 + 0.20% respectively. GHbA1C levels in these subjects were Methods: The present study was conducted on significantly higher than healthy controls (p<0.001). 25 age matched controls and 50 clinically diagnosed non-diabetic CVD patients of either sex. Conclusion:The results of the study indicate that elevated GHbA1C level is an independent risk factor Results:The GHbA1C levels in healthy controls for CVD. (n=25) and non-diabetic CVD subjects (n=50) was Abstract No: 5.22

Biochemical Markers and Coronary Occlusion in Patients Undergone Coronary Arteriography

Bhagwat VR1, Venugopal NT2, Yadav AS3 and Mane AY4 1. Dept of Biochemistry, SBH Govt Medical College, DHULE (Maharashtra), 2. Dept of Biochemistry, DY Patil Medical College, Kolhapur (Maharashtra), 3. Dept of Biochemistry, MIMSER Medical College, Latur (Maharashtra), 4. Dept of Biochemistry, RD Gardi Medical College, Ujjain (MP). Email: [email protected] Background:Cardiovascular diseases are the leaders in modern day lifestyle disorders. The risk factors are multiple and their relationships are complex. Emerging risk factors should be compared with established risk factors before its clinical usefulness can be ascertained. Objective: This study was done to evaluate the novel risk factors of coronary artery disease (CAD) with respect to the degree of coronary occlusion. Methods:The study was comprised of 33 patients who under went coronary arteriography (CAG). 9 had normal coronary, 9 had single vessel block, while 15 patients had double vessels blocks. Classic serum lipid profile included total cholesterol (TC), triglyceride, high density lipoprotein (HDL), low density lipoprotein (LDL). Novel serum lipid profile included apolipoprotein A1, apolipoprotein B and lipoprotein (a) by immuno-turbidometric assays. Abstract No: 5.23 Results: Mean TC, LDL levels and both the atherogenic indices TC/HDL and LDL/HDL showed ascending trend when compared between the groups. Mean Apo A-I in normal vessel group was highest (155 ± 11.78 mg/dl) while in double vessel block group it was lowest (135.60 ± 28.50 mg/dl). However, mean lipoprotein (a) level showed divergent trends from normal coronary vessel (5.44 ± 3.30 mg/dl), single vessel block group (13.02 ± 13.30 mg/dl) highest in double vessel block group (52.87 ± 28.42 mg/dl). Conclusion:Lipoprotein (a) is a better marker than LDL which correlated with the degree of occlusion. Higher Apo A-1 and A1/B ratio are the potential biochemical markers of anti-atherogenicity while elevated Lipoprotein (a) appears to be a dangerous atherogenic force that lead to CAD.

Cytomegalovirus and Coronary Heart Disease

Asif. A School of Health Sciences, Thalassery campus, Palayad, Kannur University [email protected] Background: The role of viruses and other infectious agents in the development of

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atherosclerosis and incident myocardial infarction and coronary heart diseases has long been an area of investigative interest. Several risk factors for coronary artery disease have been well documented including hyperlipidemia, hypertension, smoking, diabetes, a positive family history and obesity. However, these factors explain only part of attributable cardiovascular disease, and other factors must be involved.

included in the study. Serum lipid profile and determination of Anti-CMV Ig-G levels (Enzyme Immunoassay method) were performed in all the patients and the values were compared with that of normal control subjects.

Results: The serum cholesterol and Triglycerides levels in CHD patients were higher than that of normal control. The HDL-cholesterol value obtained was lower in CHD patients. The serum Objective: To Compare Ig-G levels of the CMV Anti CMV Ig-G levels were also found to be in CHD patients with normal control subjects to increased in patients with CHD. The all values find out whether the CMV plays any role in the were found to be statistically significant (P < 0.05). development of CAD. The present study was conducted on 20 CHD male patients with age Conclusion: From the findings it may be concluded that the increased level of Ig-G ranging from 40 ­ 70 year. antibodies to cytomegalovirus was associated with Methods:Twenty age matched control subjects increased risk of Myocardial Infarction and who had no past history of any cardiovascular Coronary Heart Disease. disease, hypertension of diabetes mellitus, were Abstract No: 5.24

Role of "Ischemia Modified Albumin" in the Early Diagnosis of Acute Coronary Syndromes

B Gayathri, D Vijaya, B Bini, Usha Anand., V. Aruna and CV Anand Dept. of Biochemistry, PSG Institute of Medical Sciences &Research, Coimbatore- 641004, Email: [email protected] Background: Diagnosis of cardiac ischemia in patients attending emergency departments (ED) with symptoms of acute coronary syndromes is often difficult. Cardiac troponinT (cTnT) is sensitive and specific for the detection of myocardial damage but may not rise during reversible myocardial ischemia. Ischemia Modified Albumin (IMA) has recently been shown to be a sensitive and early biochemical marker of ischemia. Methods: This study evaluated IMA in conjunction with cTnT in 150 patients presenting to the ED with acute chest pain. 10 age and sex matched healthy volunteers served as controls (non ischemic individuals). cTnT was assayed in

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Elecsys 2010 immuno analyzer (Roche Diagnostics). IMA was estimated by the method of Bar-Or et al. The values were expressed as mean ± SD and data from patients and controls were compared using students `t' test. Results: Patients were classified as ischemic, non MI ( cTnT 0.03ng/mL) and MI (cTnT >0.03ng/ mL) Values of IMA, and cTnT, were correlated with final diagnoses of unstable angina, ST segment elevation, and non-ST segment elevation myocardial infarction. The mean absorbance Units (ABSU) ± sd for control, ischemic non AMI and AMI groups were 0.3554±0.006, 0.5279±0.116, 0.6259± 0.203 respectively (p<0.0001). In the whole patient group, sensitivity of IMA at

Indian Journal of Clinical Biochemistry Supplement, 2009/24

presentation for an ischemic origin of chest pain Conclusion: IMA is highly sensitive for the early was 81%, compared with 82% of ECG and 50% diagnosis of myocardial ischemia in patients of cTnT. All three tests combined, identified 95% presenting with symptoms of acute chest pain. of patients whose chest pain was attributable to ischemic heart disease. Abstract No: 5.25

Inflammatory Markers in Arterial Prehypertension and Hypertension

Manjrekar AP, D'Souza V, Hegde A, Adhikari P· and CV Raghuveer Dept. of Biochemistry, ·Dept. of Medicine, Dept. of Pathology, Kasturba Medical College, Mangalore. Karnataka. Email: [email protected] Objective: To evaluate inflammatory markers as density cholesterol (LDL) >100mg/dL, Very low cardiac risk assessors in essential prehypertension density cholesterol (VLDL )>30mg/dL and TC/ and hypertension. HDL ratio >5. Methods: 277 undiagnosed subjects were screened and 45 subjects of age 25-60 yrs. of either gender met the criteria of prehypertention (Systolic blood pressure (SBP) 120-139mmHg; Diastolic blood pressure (DBP) 80-89mmHg) or hypertension (SBP>140mmHg; DBP>90mmHg). For this test group, an age matched normotensive control group was chosen. Waist circumference (WC) and Body Mass Index (BMI) were measured and Erythrocyte sedimentation rate (ESR), Total leucocyte count (TLC), hsC-reactive protein (hsCRP), Fibrinogen, Haptoglobin, Ceruloplasmin and fasting lipid profile were estimated. To compute the results the following cut off values were used; WC>90cms, BMI>24.9, ESR>20, TLC>7000, hsCRP>3.0mg/L, Fibrinogen >200mg/dL, Haptoglobin >150mg/dL, Ceruloplasmin >25mg/dL, total Cholesterol (TC) >200mg/dL, Triglycerides (TG) >150mg/dL, High density cholesterol (HDL) <40mg/dL, Low Abstract No: 6.1 Results: The nonspecific inflammatory markers ESR and TLC were not different in the two groups. All other inflammatory markers, dyslipidemia and obesity indices were more pronounced in the test group. Using the cut off points, the percentage values in the test/control group were; BMI-48.89/ 42.22, WC-60/40, hsCRP-28.89/6.67, fibrinogen82.22/68.89, haptoglobin-35.55/15.56, ceruloplasmin-48.89/35.55, TC-44.45/ 37.78. TG42.22/ 31.12, HDL-48.89/ 46.67, LDL-84.44/ 77.78, VLDL-42.22/ 33.33, and TC/HDL- 51.11/ 33.33. Conclusion: Inflammatory markers are independent cardiovascular risk predictors. In conjunction with altered lipid profile and metabolic indices the risk is manifold. Inflammatory markers should be considered in the hypertensive workup and anti inflammatory drugs can be included in the therapeutic management.

A Spectrum of Paraproteinemias

*K.Shanthi Naidu, **P.Srikanth, **Shailesh.S, ***M.R.Chakravarthy *Syed Omar, *Kishore, *A.Suhasini and *J.Visalasree Departments of * Laboratory Medicine, ** Consultant Pathologist and Clinical Hematologist, *** Consultant Nephrologist, CARE Hospital, Banjara Hills, Hyderabad ­ 500034, Andhra Pradesh Objective: The conventional serum protein electrophoresis (SPE) has slowly given way to

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immunofixation electrophoresis (IFE) which diagnoses a larger number of paraproteinemias and the inclusion of serum free light chain analysis, emphasizes stratification and predictive outcomes of the disease. Methods: From September 2008 ­ June 2009 324 samples, 202 males and 122 females were subjected to SPE and IFE using Beckman paragon, binding site and helena electrophoretic gels. Serum light chain analysis was done on Binding Site kit adapted to Beckman Coulter Synchron Cx9 clinical system.

Results: Paraproteinemia was detected from was 21% where as the yield from IFE was 36% (IgG kappa 64.2%, IgG lambda 4.76%, IgM kappa 14.2%, IgA kappa 4.76%). Light chain assays showed high or low ratios suggesting either suggesting an increase in either kappa or lambda, emphasizing a monoclonal pattern. Conclusion: Analysis of light chain would further evaluate patients of paraprotein, identify amyloidosis and has a major role to be used at discretion when clinical and imagiology was not diagnostic. Monitoring light chains was of prognostic significance.

Abstract No: 6.2

Effective POCT : the Need for a Broader Perspective

A.K.Srivastava Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi, India; Email : [email protected] Optimizing all the steps in the testing cycle is as crucial to the success of point-of-care testing (POCT) as the measurement process itself. The complete POCT process or testing cycle is a complex series of events that involve many parties in any clinical organization. Consultation with all the people concerned is important and has led to the formation of institution-wide Point-of-Care Coordinating committee. In recent years, the focus on developments in point-of-care (POCT) from a preoccupation with the analytical process to a broader perspective that takes into consideration other procedures which take place before and afAbstract No: 6.3 ter the measurement process. This has come after realization that success with POCT requires optimization of the many different steps that make up the total testing process or cycle. Outside forces have also played in this change of focus, particularly those concern with promoting total quality management and patient safety. As well as ensuring that POCT meets required standards on a dayto-day basis, the role of clinical scientists on the committee is to ensure that the POCT process continues to meet clinical needs through audit and, if necessary, changing testing procedures.

Biochemical Markers in Osteoporotic Hip Fractures

Dr.Meera.S **, Dr.M.Sukharaja *, Dr.B.G.Sagar # and N.Kantharaj* Mysore Medical College & Research Institute, Mysore, *Professor & HOD of Biochemistry, ** Associate Professor of Biochemistry, ***Lecturer in Biochemistry, # Professor of Orthopaedics Background: Osteoporosis is a major metabolic fracture in 40% of aging women and 15% of aging bone disease which over a lifetime results in hip men. The recognition of osteoporosis as a disease

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that leads to fracture risk, offers new insights into its epidemiology. Homocysteine is an amino acid intermediate formed during the metabolism of methionine and studies have shown it to be a strong independent risk factor for hip fracture in older men and women. The knowledge about risk factors for hip fracture in Asian population is very much limited and the mechanism(s) by which increased levels of homocysteine are associated with osteoporosis is poorly understood. Objective: To analyze the homocysteine and other biochemical parameters such as calcium, phosphorous, & alkaline phosphatase which will be compared to the Bone Mineral Density (BMD).

Methods:The present investigation involves new cases with recent history of hip fractures (n = 50) and the healthy age and sex matched controls (n= 50). The analysis of the biochemical markers is done in fasting blood samples by using fully automated analyzer. Student `t' test is used to analyze the data. This study will evaluate the role of these markers in osteoporotic hip fractures. Conclusion:The present investigation will try to throw light on the understanding of the pathophysiology of the Osteoporotic hip fracture which will help planning preventive strategies with regard to the incidence of osteoporotic hip fracture in future.

Abstract No: 6.4

Metabolic Dearrengements in Births Asphyxia Occurring as a Result of Cellular Injury.

Amrita Vamne*, B. K. Agrawal* and Jyotsana Shrivastava** *Department of Medical Biochemistry, **Department of Peadiatrics, Gandhi Medical College, Bhopal. Email address- [email protected]

Background: Birth asphyxia occurs when a body does not receive enough oxygen before, during or just after birth. Low serum calcium, glucose and magnesium values are associated with cellular injury. Objective: Present study was undertaken to

glucose by GOD ­POD method. Results: Serum calcium and glucose were found to be significantly low in asphyxiated babies compared to the control group (p<0.001). There was not a significant impact of asphyxia on magnesium metabolism. The percentage of

estimate values of serum calcium, glucose and abnormal serum calcium, glucose and magnesium magnesium on 1st, 3rd, and 5th day of life. The metabolism was found to be increased with the present study includes 50 asphyxiated and 25 severity of asphyxia in the subjects. healthy neonates. Conclusion:It is observed that hypoglycemia and Methods: Parameters included in the study are 1. hypocalcaemia are associated with asphyxiated Serum calcium by OCPC method, 2. Serum neonates, which may be due to cellular injury. magnesium by Calmagite method, and3. Serum

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Abstract No: 6.5

Evaluation of Salivary Cholinesterase and Ceruloplasmin in Children with Attention Deficit Hyperactivity Disorder

Bhavya Prabhu.K, Archana E, Priya Pai, Dr. Krishnananda Prabhu and Dr. Anjali Rao Department of Biochemistry, Kasturba Medical College Manipal, India-576104. E mail: [email protected] Introduction: Attention Deficit Hyperactivity Disorder (ADHD) is one of the most frequently diagnosed childhood psychiatric disorders. Based on various empirical estimates, symptoms of ADHD persist into adulthood in about 30-50% of the childhood cases. Studies have shown RBC and plasma cholinesterase show unique patterns of differences in patients with various neuropsychiatric disorders including ADHD as compared to controls indicating involvement of cholinergic system in these disorders. Antagonism of neuronal nicotinic acetylcholine receptors have shown to ameliorate the core symptoms of various neurological disorders associated with hyperkinesis, attention and memory loss. Patients lacking circulating serum ceruloplasmin presented with neurodegeneration. Animal experiments that showed lack of ceruloplasmin was associated with redox injury in the brain. Also copper deficiency has been implicated in neurological impairment seen in Wilson disease. Methods: After obtaining clearance from institutional ethics committee, we estimated salivary ceruoplasmin (Ravin.H.A et al) & cholinesterase (Ellman's method) from 12 controls & 9 subjects in the age group 5 ­ 12 yrs and their values were compared Results: Pseudocholinesterase showed significant increase (p< 0.001) in subjects as compared to controls.Ceruloplasmin levels showed marginal decrease(p=0.065) in subjects as compared to controls however it was not statistically significant. Conclusion: Results show the role for cholinergic system and ceruloplasmin in ADHD. Also saliva can be an effective alternative tool for evaluation. References:

1. J Atten Disord. 2005 Nov;9(2):382-3 2. Psychiatr Pol. 2002 Nov-Dec;36(6 Suppl):133-41 3. Curr Opin Neurol. 2006 Dec;19(6):613-8 Objective:: To estimate salivary ceruloplasmin 4. Biochem Soc Trans. 2008 Dec;36(Pt 6):1277-81 and cholinesterase in children with ADHD & to 5. J Neurol Neurosurg Psychiatry. 2009 May;80(5):524-7 compare their respective values in controls. 6. Pediatr Neurol. 2006 Oct;35(4):284-6 Abstract No: 6.6

Cheek Cell Epithelium: a Non Invasive Specimen for Measurement of Fatty Acid Intake

Ransi Ann Abraham, Lakshmy Ramakrishnan ,Ruby Gupta, Smita Golandanz, Rajinder Parshad, V K Bahl, Ambuj Roy, R M Pandey, Dorairaj Prabhakaran and Kolli Srinath Reddy Department of Cardiac Biochemistry, All India Institute of Medical Sciences, New Delhi

[email protected]

Objective:The intake of fat, an essential macronutrient of diet vary widely among

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individuals and changes over time. Traditional dietary assessment methods are prone to errors and

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bias; hence an objective measurement is important to assess intake. Technical advancements have made it possible to study fatty acid even from small tissue samples; hence we have explored the suitability of using cheek cell epithelium as a non invasive tissue for assessing dietary fatty acid intake. Methods: Blood and cheek cells were collected from 17 subjects in fasting state at the same time. Dietary data was collected through 24 hrs recall and food frequency questionnaire. Lipids were extracted from cheek cell and serum; fatty acids converted to methyl esters and were run on GC. Fatty acids were identified by comparing their retention time with known standards. Dietary fatty acid was computed from the food composition tables.

and 33.06 (4.66) respectively, MUFA was 21.27 (8.75) and 24.76 (3.74) respectively and PUFA was 12.76 (6.04) & 24.52 (3.34) respectively. Palmitic, stearic and oleic acid were the major fatty acids identified in cheek cell and serum. There was a significant correlation between MUFA (r = 0.35) and arachidonic acid (r = 0.46) values obtained between serum and cheek. The saturated fatty acids in cheek cell correlated significantly with dietary saturated fatty acid (r = 0.52). A correlation of 0.48 & 0.51 was observed for linoleic and total PUFA in serum and diet respectively. The distribution of fatty acid varied between serum and cheek cell. Serum was richer in PUFA which reflected the dietary intake of the subject, while cheek cell reflected the saturated fatty acid intake which are stored as fat.

Conclusion:In conclusion saturated fatty acids in Results:The % mean (SD) total saturated fatty cheek can be a good non invasive biomarker for acids levels in cheek and serum were 35.13 (8.06) dietary saturated fatty acid intake. Abstract No: 6.7

Coagulation Deficiencies in a Referral Laboratory

*K.Shanthi Naidu, **P.Srikanth, **Shailesh.S, ***D.Ramesh, #D.Ramana, *B.Raghavendra, *B.Goud and *B.Siva Mallika Departments of * Laboratory Medicine, ** Consultant Pathologist and Clinical Hematologist, *** Consultant Peadiatrician*, ** & *** CARE Hospital, Banjara Hills, Hyderabad ­ 500034, Andhra Pradesh, # Rainbow Hospital, Hyderabad, Andhra Pradesh Objective: Coagulation studies need critical assessment of standard operative procedures and quality control. Evaluation of either congenital or acquired deficiency should always be done where technical expertise is high and hence no hesitation to a higher referral center should be debated. Results: Of 1000 referrals to our hospital laboratory 252 were abnormal: The following were identified; factor VIII 173 (68.6%), factor IX 43 (17%), factor V 5 (1.98%), factor X 12 (4.7%), factor VII 4 (1.6%), factor XIII 2 (0.8%), fibrinogen 5 (2%), combination of factor V and Methods: Coagulation studies pertaining to a VIII 2 (0.8%), inhibitors to factor VIII and IX 6 (2.4%). routine screen of prothrombin time, activated partial thromboplastin time, fibrinogen, thrombin Conclusion: Appropriate clinical and family time, factor XIII, platelet and pheripheral smear history narrows down the diagnosis of deficiency. studies were done on CA 50, Stago ST and Sysmex Mixing experiments further help in aiming at a 4. Bleeding time by modified Ivy's technique, particular assay, cutting down cost to the patients where necessary. Mixing experiments and factor and highlighting efficiency of the laboratory in assays by commercial reagents of Transasia, Stago understanding the sequence of assays to be done. and Tulip Diagnostics

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Abstract No: 6.8

Coagulation Studies in Neonatal Critical Care

*K.Shanthi Naidu, *P.Srikanth, **Shailesh.S, ***D.Ramesh, #D.Ramana,*B.Raghavendra, *B.Goud, *B.Siva Mallika and *S.M.Ismail Departments of * Laboratory Medicine, ** Clinical Hematology, *** Peadiatric*, ** & *** CARE Hospital, Banjara Hills, Hyderabad ­ 500034, Andhra Pradesh, # Rainbow Hospital, Hyderabad, Andhra Pradesh Objective: Altered hemostatic parameters both coagulation and platelets in neonates is a call for immediate attention, being a critical issue. Understanding the normal reference range in this group is essential prior to labeling a disorder. Methods: 150 samples over a period of 2 years studies for peripheral smear, platelets, prothrombin time(PT), activated partial thromboplastin time(APTT), fibrinogen and thrombin time(TT) were assayed on Sysmex analyzer, CA 50 and Stago ST 4. Factor XIII was done by urea clot lysis time. Results: Abnormal PT and APTT were seen in 38%, low fibrinogen levels in 10.8%. Factor XIII Abstract No: 6.9 abnormality was mainly due to low fibrinogen levels preventing a clot formation. Hemorrhagic disease of the new born was 34%. All studies were done on follow up to confirm this. Factor VIII and factor IX deficiencies were retested over a period of 6 months to one year for confirmation unless the history and clinical presentations identified a congenital deficiency. Conclusion: Close monitoring of neonates for coagulation studies is related to the process of phlebotomy and the specific reference range very different from the adult. Deviation from this protocol would mislead in over diagnosing simple variability by a large protocol of workup.

A Case of Xanthochromia

Ashish Sharma, N. Haridas, S. Ghosh, Hitesh Shah ,Kiran and Uday Department of Biochemistry, P.S. Medical College,Karamsad Anand, Gujarat Sub arachnoid hemorrhage detection is very crucial in patients. SAH can develop due to varied etiologies. A computerized tomography (CT) scan is positive in up to 98% of patients with SAH presenting within 12 h, but is positive in only 50% of those presenting within first week. Here we are presenting a case having a history of road traffic accident, came to our hospital after three weeks of RTA with CT finding inconclusive at admis190

sion. Patient had continuous headache and neuropsychiatric symptoms. Biochemical analysis of CSF revealed increased level of bilirubin (xanthochromia) which was suggestive of Sub arachnoid hemorrhage. In this way biochemical CSF analysis is important in diagnosis and therapeutic selection of patients for angiography and preventative surgery.

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Abstract No: 6.10

Abstract No: 6.11

The Potential Significance of Procalcitonin and Interleukin-6 in Prediction of Survival in Patients with Sepsis

S. Das, S. Bhargava, A. Manocha, M. Kankra and L.M.Srivastava Department of Biochemistry, Sir Ganga Ram Hospital, New Delhi, India Objective: Appropriate treatment of sepsis relies on early and correct diagnosis and contributes to reduced morbidity and mortality. Procalcitonin (PCT) is a proposed specific marker of the systemic inflammatory response to infection and interleukin-6 (IL-6) is a proinflammatory cytokine induced by many mediators. Our objective is to determine the relationship between levels of serum

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PCT and IL-6 in patients with sepsis and their prognostic outcome. Methods: ICU patients with suspected infection and fulfilling at least two criteria of systemic inflammatory response syndrome were studied. PCT and IL-6 were measured on admission (0 hour), 24 hours and 72 hours and their levels compared in surviving and non surviving patients.

non survivors as compared to survivors. However, PCT levels decreased significantly (p=0.003) in 58% of surviving patients over a period of time (0-72 hours) as compared to 29% in non surviving group. The difference in IL-6 levels between survivors and non survivors was not statistically significant.

Conclusion: Significant decreasing levels of PCT as compared to non significant decrease of IL-6 in Results: The median value of PCT and IL-6 were survivors with respect to non survivors were significantly high (p=0.000 - 0.032) in non associated with a higher probability of survival. surviving patients as compared to patients who Hence, time dependent monitoring of PCT survived. The acute physiology and chronic health concentration might provide an earlier indication evaluation and sequential organ failure assessment on the chances of survival. scores were also significantly (p=0.000) high in Abstract No: 6.12

Immunological Studies on Glycated and Glycoxidated LDL: Implications in the Pathogenesis of Various Diseases

Salman K A1, Alam R1, Arif N2 and Khan S1 Department1 of Biochemistry, JN Medical College, Aligarh Muslim University and Department2 of Chemistry, Z H College, New Delhi. Background: The entry of glucose into the brain and peripheral nerve tissue, kidney, intestine, lens and RBC does not depend on insulin action. During hyperglycemia, intracellular level of glucose in these cells is high. This promotes nonenzymatic attachment of glucose to protein molecules (Protein Glycation). Glycation of proteins leads to the formation of AGE's (Advanced Glycation End product) of diverse molecular structure and biological functions. AGE cross-links body proteins such as tissue collagen or a nerve protein-myelin, lens crystalline. AGE formation stiffens the extracellular matrix and decreases the elasticity of, for instance the arterial wall. Their formation also affects the function of endothelial cells, phagocytes (macrophages) and smooth muscle cells in the wall of blood vessels. Through these mechanism AGE may contribute to the development of the late complication of diabetes other vascular diseases, aging and rheumatoid arthritis.

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Objective: The basic objective of this study is to evaluate the level of glycated and glycoxidated LDL in the sera of patients with vascular diseases and diabetes. Methods: Commercially available LDL was modified by glucose and peroxynitrite generated by SIN-1. The degree of LDL modification was assessed by various physical and biochemical parameters. Results: Serum autoantibodies against native, glycated and glycoxidated LDL was evaluated in patients of cardiovascular diseases and diabetes. The results showed the presence of high levels of autoantibodies against glycoxidated LDL as compared to native and glycated LDL. Conclusion: Such research also will increase the understanding of how AGES cause pathology and the extent of AGE involvement in the various complications of diabetes and vascular diseases.

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Abstract No: 6.13

Critical Call Back ­ An Experience in a Tertiary Care Hospital

Dr. Mohammad Ibrahim1, Maj Dr. Ikramul Islam (Retd) 2, Prof (Dr.) Tareak-Al-Nasir3, and Lt.Col Dr. Debashish Saha4 1.Senior Registrar, Clinical Biochemistry, Lab Medicine Department Apollo Hospitals Dhaka, Bangladesh. 2.Senior Registrar, Clinical Pathology and Hematology, Lab Medicine Department Apollo Hospitals Dhaka, Bangladesh. 3.Senior Consultant and Coordinator Lab Medicine Department Apollo Hospitals Dhaka, Bangladesh. 4.Classified specialist in Pathology CMH Bogra Cantonment, (ExConsultant, Clinical Biochemistry, Lab Medicine Department Apollo Hospitals Dhaka). Bangladesh Critical value reporting originally was highlighted by Dr. Lundberg, who defined a critical value as a result suggesting that the patient is in imminent danger unless appropriate therapy is initiated promptly. Reporting of laboratory critical values has become an important issue for patient safety as described in the National Patient Safety Goals of the Joint Commission on Accreditation of Healthcare Organizations (JCAHO).The recent focus on patient safety has brought increased attention to the issue of laboratory critical value reporting. Critical value reporting parameters may be considered an important laboratory outcome measurement because they reflect clinical effectiveness, patient safety, and operational efficiency. For the critical value reporting process to be effective, the organization must understand and address the variables involved in the process. Herein, Abstract No: 6.14 we report the results of an analysis of 4260 consecutive laboratory critical values from July 2007 to December 2008 at our institution, a Joint Commission International accredited Tertiary care hospital to understand the scope of critical value reporting and identify opportunities for process improvement. We evaluated critical value reporting by parameter, laboratory specialty, clinical care area, time of day, and Turn around Time. Factors leading to delays in critical value reporting are identified, and we describe approaches to improving this important operational and patient safety issue. We are proud to be the pioneer in Bangladesh to introduce the system of Critical Call Back in Lab medicine of Apollo Hospitals Dhaka which was highly appreciated by the survey team of Joint Commission on Accreditation of Healthcare Organizations (JCAHO) during our accreditation process

Rare cases of Pagets Disease of the Bone in India: Biochemical and Clinical Evaluation and Follow-up in 2 Extreme Presentations

Dr CS Muralidhara Krishna ,Dr H L Vishwanath ,Dr S Sanjay, Dr Tejeswini Deepak, Dr A Sharda and Dr SS Srikanta Endocrinology and Diabetes Study Group, Samatvam and Jnana Sanjeevni Medical Center, and Dept Biochemistry, Bangalore Medical College, Bangalore, India Paget disease is a localized disorder of bone remodeling that typically begins with excessive bone resorption followed by an increase in bone formation. Approximately 70-90% of persons with Paget disease are asymptomatic and detected based on the incidental finding of an elevated serum alkaline phosphatase (SAP) level or characteristic radiographic abnormality However, a minority of affected individuals experience various symptoms, including bone pain (the most common symptom), secondary osteoarthritis, bony deformity, and neurologic complications.

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Although the etiology of Paget disease is unknown, both genetic and environmental contributors have been suggested. Ethnic and geographic clustering of Paget disease is well described. An environmental trigger for Paget disease has long been considered but never proven - viruses. Paget disease is estimated to occur in 1-3% of individuals older than 45-55 years and in up to 10% in persons older than 80 years ­ Europe. Paget disease is very rare in Asian countries, especially India. Bisphosphonates (eg alendronate 40 mg/d PO for 6 mo) are first line therapy - analogs of pyrophosphate, act by binding to hydroxyapatite in bone matrix, thereby inhibiting the dissolution of crystals. Case 1: DS, 53 year male, was found (year 2008) to have unexplained asymptomatic elevation of SAP 625, 908, U/L (53-128), during a routine annual diabetes health review; Pagets disease ­ polyostotic was confirmed by isotope bone scan Abstract No: 6.15

(skull, vertebra, pelvis, femur, tibia, scapula, sternum) and radiology; he was treated with the initial cycle of alendronate ; during and after therapy serial SAP levels decreased to 786, 521, 299 and 136U/L. Case 2: BN,63 year female presented (year 2002 ) to an orthopedic surgeon with severe L iliac bone pain, leading to x ray studies - lytic bone lesions, isotope bone scan, bone biopsy and a histological diagnosis of "malignancy" ­ metastases from an unknown primary - erroneous. Review by endocrinologist established the diagnosis of Pagets diseases ­ polyostotic (skull, vertebra, pelvis, femur), SAP level of 1343,954,835. She received two 6 months cycles of alendronate therapy in years 2003 and 2007 (Serial SAP; Year 2003=130,168;Year 2004=232;Year 2005=322,Year 2006=339 ,Year 2007=575,Year 2009=65).

Role of Acute Phase Proteins in the Prognosis of Head Injury

P Sumathi and M K Balasubramanian Department of Biochemistry, SRM Arts and Science College, Kattankulathur ­ 603 203, Tamilnadu Objective: Assessing the patients' response to stress in terms of acute phase proteins maybe of great value in determining the prognosis head injury patients and also for providing possible new therapeutic management protocol. Methods: The present study was based on 103 prospectively selected patients admitted to the male head injury intensive care unit, institute of neurology, government general hospital Chennai. Blood samples were drawn between 10 am and 11 am by venipuncture on admission (within first 24 hours of trauma) and on every alternate day during the first week and once in a week during the second and third week. The control group had

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52 healthy volunteers .clinical outcome was assessed based on the Glasgow outcome scale (GOS) and the patients were classified into three groups as group1: patients with good recovery, group2: patients with moderate or severe disability and group3: patients in persistent vegetative state or death. Radial immunodiffusion (RID) assay was used for c-reactive protein (CRP), ceruloplasmin (CS) and transferrin estimation. Albumin was assayed using dye binding method. Results: The patients with head injury had a profound acute phase response which lasted throughout the 21 day study period. Increasing level of CRP was noted up to fifth day, after which

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it decreased slowly but still remained significantly higher than control throughout the study period. A similar trend was observed with CP levels in which peak levels were recorded during the second week. The concentration of transferin fell rapidly after head injury and the levels remained below normal in all the three groups throughout the three weeks time. Using a stepwise regression analysis a prognostic index was derived. GCS predicted the outcome of 61.1 % of patients correctly whereas in conjunction with T4, TCS, TRP and

CP the prognostic ability improved to 86.5 %. Stepwise multiple discriminant analysis yielded three prognostic indeces I, II and III for group 1, 2 and 3 respectively with a correct prediction average of 93.2. Conclusion: From the discriminant function analysis it was found that acute phase reactant such as CP and CRP may be of paramount help to distinguish group 2 patients from others

Abstract No: 6.16

Biochemical Profile of Cord Blood in Normal Pregnancy and Pregnancy Induced Hypertension.

Miss D.Prathima , Dr. U.B.Vijaya lakshmi and Prof. Dr. K. Ambika Devi Dept. of Biochemistry ,ASRAM Medical college , Eluru, A.P; Email : [email protected] Background : Pregnancy induced hypertension (PIH) is a significany health problem and is the leading cause of maternal and perinatal mortality and morbidity. Low birth weight and prematurity is common in such cases. Objective: This study was designed to compare certain biochemical parameters in cord blood of children born to mothers with PIH and to mothers with normal pregnancy. Methods: For this purpose 10ml of cord blood during delivery was collected from mothers attendingobstetrics department at ASRAM medical college,Eluru.30 samples were collected from mothers with uncomplicated pregnancy and 50 samples from the mothers with PIH (maternal age matched).The 50 samples include group A20 mothers with pre-eclampsia and group B-30 mothers with gestational hypertension.The biochemical parameters measured were

glucose,urea,total proteins,total cholesterol,calcium & magnesium. Results: It was observed that all the parameters except for total cholesterol and calcium were significantly lower(p<0.00001) in the cord blood of babies born to mothers with PIH.The total cholesterol and calcium were significantly higher in the same cases (p<0.001).On comparison of parameters of cord blood in babiesweighing > 2.5kg in cases vs controls,there was significant decrease in all the parameters in the former. Conclusion: So the neonates born to mothers with PIH must be screened carefully to prevent mortality and morbidity due to low levels of the important biochemical parameters like glucose and magnesium.Neonates weighing >2.5kg are also at similar risk

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Abstract No: 6.17

Comparision of Plasma Phosphdiesterase Activity in Premenstrual and Menstrual Period.

Rajyalakshmi M, Indu R Warrier, Dr. Krishnananda Prabhu and Dr. Anjali Rao Department of Biochemistry, Kasturba Medical College Manipal, India-576104. E mail: [email protected] Background: Both menstruation and pregnancy are inflammatory conditions that cause a degree of physiological ischemia-reperfusion. Contractility of uterine smooth muscle is essential for the cyclic shedding of the endometrial lining and also for expulsion of the fetus during parturition. Cyclic nucleotide phosphodiesterases (PDEs) are a diverse enzyme group with multiple regulatory properties and wide tissue distribution. Such activity includes cyclic adenosine (cAMP) and guanosine monophosphate (cGMP) breakdown. Cyclic nucleotide accumulation has been implicated in dilatation of intact tonic spiral arteries uterus and in relaxation of myometrium. Isoforms of the PDE4 family are shown to be involved in smooth muscle contraction and inflammation. PDE4 inhibitors have been shown to prevent inflammation-induced preterm delivery. age 21 yrs in the premenstrual period and on first day of menstruation. Their respective premenstrual and menstrual levels were compared. Results: We observed that there is a significant increase (p <0.05) in the phosphodiesterase levels in the menstrual period as compared to their respective values in the premenstrual period. Conclusion: Our results show that there is a significant change in levels of plasma phosphodiesterase indicating their possible role in uterine contraction. References:

1. Am J Obstet Gynecol. 2004 Jan;190(1):268-74 2. Biol Reprod. 2001 Mar;64(3):857-64 3. Ginekol Pol. 2000 Aug;71(8):789-93 4. BMC Pregnancy Childbirth. 2007 Jun 1;7 Suppl 1:S12 Methods: Plasma phosphodiesterase were 5. J Immunol. 2007 Jan 15;178(2):1115-21 estimated in healthy female participant students 6. BMC Pregnancy Childbirth. 2007 Jun 1;7 of Kasturba Medical College, Manipal with mean Suppl 1:S10 Abstract No: 6.18

Serum Calcium, Phosphorus and Alkaline Phosphatase Levels in Preterm Babies.

Sarika Singh Chauhan*, B. K. Agrawal* and Rashmi Dwivedi** *Department of Medical Biochemistry, **Department of Paediatrics, Gandhi Medical College, Bhopal. Email address- [email protected] Objective: Preterm is defined as a baby with a gestation of less than 37 completed weeks. Preterms have low serum calcium and phosphorus levels with increased serum alkaline phosphatase

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activity. In this study serum calcium, phosphorus and ALP activity were measured in preterm babies.

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Methods: The present study comprised of 75 preterm babies. Of which 25 were of 28-30 weeks, 25 were of 30-32 weeks & remaining 25 were of 34-36 weeks (Controls) of gestational age. In these patients following investigations were done to assay serum calcium, phosphorus levels and serum alkaline phosphatase activity.1. Serum calcium by OCPC method. 2. Serum phosphorus by Modified Metol method, and 3. Serum alkaline phosphatase by Kinetic p-NPP method.

increased (p<0.001) at 28-30 weeks as compared to 34-36 weeks (Controls), but serum calcium & phosphorus levels were found to be insignificantly decreased whereas serum ALP activity were found to be insignificantly increased at 28-30 weeks as compared to 30-32 weeks of gestational age in preterm babies. The correlation between serum calcium & phosphorus were found to be positive at all gestational ages, whereas Serum ALP is inversely correlated with serum calcium & phosphorus at all gestational ages. It can be Results: Serum calcium & phosphorus levels were concluded that high serum ALP activity & low found to be significantly decreased and Serum serum calcium & phosphorus levels are associated ALP activity was found to be significantly with preterm babies. Abstract No: 6.19

Study of Effect of Phototherapy on Neonatal Hyperbilirubinemia

M Vijay and M Malathi. Department of Biochemistry, Father Muller Medical College Mangalore, Karnataka. Email:[email protected] Objectives 1. To study effect of phototherapy on neonatal hyperbilirubi-naemia. 2. To compare effect of photo-therapy on term and preterm neonates. Methods: The present study was conducted at between January 2009 and may2009 in Father Muller Medical College, Mangaore. Patient's data were collected from NICU phototherapy Unit of department of paediatrics. 42 cases were identified out of which 25 were term and 17 were preterm babies. Phototherapy was initiated by paediatrician according to clinical assessment, lab results and reference charts. Mode of phototherapy is continuous. Blood level of bilirubin; (total, unconjugated and conjugated in mg/dl) were analyzed by auto analyzer Olympus-AU 400. Levels of bilirubin recorded daily in the laboratory of department of Biochemistry. Effect of phototherapy on neonatal unconjugated hyperbilirubinaemia was studied. Data were analysed by paired "t" test. The values of significance were evaluated with `p 'values. The difference were considered significant at p<0.05. Results: Phototherapy is effective treatment for neonatal jaundice and the mechanism-is well defined. The therapeutic effect depends on the light energy emitted in the effective range of wavelengths, the distance between the lights and the infant, and the amount of skin exposed, as well as the rate of hemolysis and in vivo metabolism and excretion of bilirubin. In our study phototherapy was effective in term neonates, as decreasing level of both total and unconjugated bilirubin is significant when compared daily. However phototherapy was effective in preterm neonates, but decreasing level of both total and unconjugated bilirubin is not significant because preterm neonates take longer duration. Conclusion: Jaundice is the most common clinical symptom and sign in neonatal period, arising from either overproduction or under secretion of bilirubin. Almost all neonates (60% Term and 80% Preterm) will have bilirubin greater than 5 mg/ dl in the first week of life and about 6% of term babies will have levels exceeding 15 mg/ dl. Due to

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immaturity of the enzymes involved in bilirubin conjugation. High levels of unconjugated bilirubin are toxic to the newborn ­ due to its hydrophobicity it can cross the blood-brain barrier and cause a type of mental retardation known as kernicterus. If bilirubin levels are judged to be too high, then phototherapy with UV light is used to convert it to a water soluble, non-toxic form. Management of jaundice is directed towards Abstract No: 6.20

reducing the level of bilirubin and preventing CNS toxicity. Bilirubin in the skin absorbs light energy, which by photo-isomerization converts the toxic native unconjugated 4Z, 15Z-bilirubin into the unconjugated configurational isomer 4Z, 15Ebilirubin. The latter is the product of a reversible reaction and is excreted in bile without any need for conjugation.

SMS Tube : A Novel Way of Giving Specimen Collectors 24/7 Mobile Access to Our Specimen Requirements Database

T F Hartley1, W. Bowling, Sharon Fong, A, Gibb, K Hen, B Hitchens, Renee Twynam-Perkins, K Wilson, J Zhang and Nicole Herbert, 1 Quality Manager, Pathology Services, Royal Hobart Hospital, Hobart, Tasmania, Australia. School of Computing and Information Systems, University of Tasmania, Hobart, Tasmania, Australia. Email: [email protected]

Objectives: Pathology Laboratories typically receive many repetitive telephone calls from Doctors, Nurses and Phlebotomists enquiring about what tube they need to use to collect a blood sample for various tests. In our practice these clients do have access to an online web database but many claim that they do not have convenient access to a PC to use that. This project involved the development of a mobile phone SMS based system which we anticipate will reduce the amount of time `wasted' on handling these calls. Method: Our existing specimen requirements database, currently viewable at www.centrepath.dhhs.tas.gov.au has been ported to an SMS database query server for this project. Typically our clients interact with this system as follows Specimen collection staff cannot recall what tube to use when they are just about to collect a blood sample for cholesterol testing so they create a

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quick SMS query message on their mobile phone eg `cholesterol' They then send this SMS message to our SMS database server Our system uses a unique program to query the database and respond `Cholesterol : 4ml blood, red top tube, patient should be fasting for 12 hours. Results same day.' If the SMS text from the caller is not immediately recognisable eg Creat then the system responds with a few options `Did you mean Serum Creatinine Urine Creatinine Creatinine clearance None of the above Respond by SMSing us the closest number from this list'

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If the SMS text query is totally unrecognisable then, in Release 2 of the software, it will alert our Specimen Reception staff who will then phone back the caller. Results: The Release 1 of the software has been shown to be capable of handling SMS query messages via a GSM modem. It has been able to analyse the content of the message to find key acronyms and has used these to search the database. Once the acronym has been matched then the relevant information has been compiled and formatted to the size of a standard SMS text message (160 characters) and this has been sent back to the mobile number from which the request originated. Basic session management has been implemented as part of Release 1. The system has opened new sessions for each new SMS query that it has received. If a query has been matched exactly by the database then the system has responded with the name of the matching test and the relevant tube information. The session has then been closed. If there have been multiple matches for the query, Abstract No: 6.21

the system has responded with a simple "Did you mean?" and provided a list of the possible alternatives, with the final one being "None of the above". When the user has chosen one of the options, then the corresponding database entry has been sent. The session has been kept open until the user replies with an option. If the user has chosen the last option, "None of the above", then the query and the user details have been logged, but Pathology has not been notified in this Release 1 version. This has closed the session. Any open sessions have been terminated after 10 minutes if the user has not responded. Unresolved queries have been notified via an SMS from the system to the user. Conclusion: The basic functionality has been shown to be achievable and by the time of presentation delegates to ACBICON will be able post SMS queries from their own mobile phones into our system in real time. In addition the various system management reports on usage statistics will be demonstrable

Storage Stability of Glycosylated Hemoglobin in Red Blood Cells and Dried Blood Spots

Ruby Gupta, Ramakrishnan Lakshmy, Dorairaj Prabhakaran and Kolli Srinath Reddy Department of Cardiac Biochemistry, All India Institute of Medical Sciences, New Delhi-29. [email protected] Background: Glycosylated hemoglobin (GHb) is an indicator of long term management of plasma glucose; therefore it is essential that the measurement of GHb be easier, especially for distance monitoring in health care, population screening and clinical trials. For the purpose of distant monitoring and research, the stability of samples collected and their appropriate storage is of major concern. Objective: Keeping these points in view, we have estimated GHb in RBCs as well as dried blood spotted on filter paper (DBS) and compared with Whole Blood (WB).

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Methods: To study the effect of storage, accounting for the lapse of time during transportation of samples from one place to another, GHb was estimated in RBCs and DBS after 30 days of storage. Twenty samples were collected from the routine collection counter. Blood spots were prepared on Whatman filter paper and dried at room temperature. These DBS were then stored in zip-lock poly bags. An aliquot of whole blood was separated for analysis of GHb. The rest of the blood was centrifuged at 2500 rpm. Plasma was discarded and RBCs were stored for the estimation of GHb. Analysis was done on day 1 and day 30. Abstract No: 6.22

Results: The mean GHb measured in WB, RBCs and DBS were 6.50 ±1.44 %, 6.10 ±1.44 % & 6.70 + 1.25 % respectively. The correlation coefficient (r) between WB & RBCs was 0.996 & between WB and DBS it was 0.986. The mean GHb in RBCs were 7.07 ± 1.95 % on day 1 and 7.0 ± 1.75 % on day 30 (r = 0.983). The mean values for DBS was 7.21 ± 1.55 % on day 1 and 7.36 ± 1.42 % on day 30 (r = 0.950). Conclusion Glycosylated Hemoglobin in RBCs and Dried Blood Spots showed good correlation with the values in Whole Blood and were also stable on storage up to 1 month and therefore can be used in population screening and clinical trials.

Data Analysis of Biochemical Values in a Polyclinic and Diagnostic Centre in Dumdum, Kolkata

Dr.Aditi Sen (Sarkar), Sanchari Halder, Anirban Ganguly, Mononjay Banerjee, Ramkrishna Bhattacherjee and Dr. Papia Mukherjee Dept of Pathology, Theism Ultrasound Centre, 14-B, Dumdum Road and 3A/13 B K Paul Avenue, Kolkata 700030; Email: [email protected]

Methods: The present work involves a systematic study of blood samples sent to a NABL accredited Clinical Biochemistry Laboratory to obtain normal and abnormal percentages of various test values. The common tests performed on these samples are for plasma glucose, serum urea, creatinine, uric acid, cholesterol, liver function tests, lipid profile, electrolytes and hormonal analysis. The daily average of samples analysed is 300 and the tests performed is 600. The instruments used are Cobas Integra 400, Elecsys 2010, Roche 9180 ISE and Cobas 6000. The present analysis considers an annual input of 1,59,894 tests. Results: Plasma glucose was normal for 66%, hypoglycemia occured in 0.9% and hyperglycemia was found in 33.1% cases. Urea was normal in 92%, high in 7.8% and low in 0.2% cases. Uric acid was normal in 87% cases while 13% had high

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values. Bilirubin value was normal in 97%, high jaundice was observed in 3% of which 5% cases were HbsAg positive. Cholesterol in 90.7% cases was normal and 9.3% cases were high. Triglyceride was normal in 78% with 22% cases being high. Thyroid function tests were normal in 81.2%, hyper in 3.6% and hypo in 15.2% cases. Electrolyte imbalances were observed in 17.2% cases of which 60% were aged above 65yrs. Conclusion The inference is that almost 10% cases of urea, uric acid, bilirubin and cholesterol are abnormal; tryglycerides, thyroid function and electrolytes were abnormal in almost 20% cases and hyperglycemia had the highest occurrence. Increasing tendency of these trends may be due to stresses, pollution in the city and also lifestyle changes caused by globalization.

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Abstract No: 6.23

Evaluation of Salivary Magnesium and Protein Thiols in Children with ADHD

Archana E, Bhavya Prabhu K, Priya Pai, Dr. Krishnananda Prabhu MD, Dr. Anjali Rao, and Dr. Revathi P. Shenoy Department of Biochemistry, Kasturba Medical College, Manipal, India-576104. E-mail: [email protected] Background: Attention-deficit hyper-activity disorder (ADHD) is the most prevalent behavioral disorder in children and the pathophysiology remains obscure. Ionic magnesium (Mg(2+)) depletion has long been known to cause hyperexcitability with convulsive seizures in rodents, effects that have been reversed by treatment with magnesium. A positive influence of magnesium in the prevention and treatment of hyperactivity in children is frequently raised in literature. The magnesium levels were found to be moderately decreased in plasma and erythrocytes, and the Mg2+ -ATPase activity was also reduced in ADHD children. Oxidative stress may be a common pathogenic mechanism underlying many major psychiatric disorders, as the brain has comparatively greater vulnerability to oxidative damage.The levels of protein thiols in the body indicate antioxidant status. protein thiol by Ellman's method and Magnesium by colorimetric method. Results: Salivary thiols showed a significant increase in subjects (p < 0.001) as compared to controls and magnesium showed significant decrease in subjects (p < 0.01) as compared to controls. Conclusion: This result substantiates the role of oxidative stress and magnesium in pathophysiology of neurological disorders. Also saliva can be an effective alternative tool for evaluation. References;

Magnes Res. 1997 Jun;10(2):143-8 J Am Coll Nutr. 2004 Oct;23(5):545S-548S Int J Neuropsychopharmacol. 2008 Sep;11(6):85176 Objective: Our study was undertaken to quantify Prostaglandins Leukot Essent Fatty Acids. 2008 the salivary levels of magnesium and protein thiols Jul-Aug;79(1-2):47-53 in children with ADHD and compared with the J Psychiatry Neurosci. 2007 Nov;32(6):435-8 age matched healthy controls. Methods: Saliva from 9 children with ADHD as well as 12 healthy children was investigated for Abstract No: 6.24

Estimation of Creatinine in Sweat

Shivaraj Gowda, Prakash Desai, Avinash A K Math, Sonal Vernekar, Shruthi Kulkarni, Vinayak Hull and Mahantesh Ilkal Department of Biochemistry, J. N. Medical College, Belgaum ­ 590010; E-mail: [email protected] Background: Creatinine is a metabolic product constant rate by the body. Creatinine being a of muscle metabolism and is produced at a nitrogenous compound is primarily eliminated

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from the body by the kidney via the glomerular filtration mechanism. Creatinine is also excreated through sweat. There are few literatures showing creatinine concentration in sweat. The aim of our study was to determine creatinine concentration in sweat in normal individuals.

and sweat creatinine was estimated by alkaline picrate method. Results: 0.029 mmoles/L is the mean concentration of creatinine in sweat has been obtained.

Methods: The procedure for the collection of Conclusion: The creatinine concentration sweat: Arms of 10subjects were washed and full obtained in sweat in our study is consistent with arm were wrapped with thin polythene bag. the results shown by Chien-Tsai Huang et al. Samples were collected after 30 min of exercise Abstract No: 6.25

Reference Range for Serum Total Alkaline Phosphatase in Healthy Individuals

Kiran Kumar B1 , Nalini Ganesan 2 and Anbazhagan M 3 1 PG Student, 2 Professor, 3 Chief Biochemist, Department of Biochemistry, Sri Ramachandra Medical College & Research Institute, Sri Ramachandra University, Porur, Chennai - 600 116; Email address: [email protected] Background: Reference value is important in establishing a scientific basis for clinical interpretation of laboratory data. It is the result of certain type of quantity obtained from a group of individuals corresponding to a state of description. Alkaline Phosphatase (EC 3.1.3.1) hydrolyses organic phosphates at alkaline pH and the serum Alkaline Phosphatase has diagnostic significance in liver and bone diseases. The level of this enzyme is influenced by the age and sex. In order to understand the concept of reference values and also to learn the process of establishing the reference range, this study was undertaken. `informed-consent'. Two ml of random blood sample was collected and serum was separated and used for the enzyme assay. Serum total Alkaline Phosphatase was assayed using commercial (Siemens) kit. In this kit, p-nitrophenyl phosphate was used as an artificial substrate. The results were expressed as U/L. The statistical analysis was done using students `t' test.

Results: The Normal range obtained in this study is as follows: Group 1 (5-20 years): 78-152 U/L, Group 2: (21-30 years): 48 - 132 U/L, Group 3 (31-40 years): 53-117 U/L, Group 4: (41-50 years): Objective: To determine the reference range for 44-140 U/L. Our result shows that maximum serum total Alkaline Phosphatase in different age Alkaline Phosphatase activity is in the age group 5 - 20 years. groups of healthy individuals. Methods: Sex matched healthy subjects of different age groups ( each consisting of 30 volunteers) required for our study were selected from the students and staff members of our Medical College & Hospital after getting their

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Conclusion: The present study has enabled us to understand the factors affecting the reference value, criteria to be considered during sample collection, analysis and the importance of reference ranges.

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Abstract No: 6.26

Quality Control Evaluation using Quality Indicators - Experience of a Clinical Laboratory in Delhi

A Manocha, S Bhargava, M Kankra, S Das and LM Srivastava Department of Biochemistry, Sir Ganga Ram Hospital, New Delhi, India Background: Competence of clinical laboratories has been increasing in India due to quality management and accreditation. This is assured by producing accurate and precise data for patient care. Establishment of quality indicators allows detection and quantification of problems, which are essential for complete quality checks. Internal Quality Control (IQC) is based on daily monitoring of test procedures performed in the laboratory. External Quality Assurance Scheme (EQAS) refers to objectively checking laboratory results through an external agency and includes inter ­ laboratory comparisons. EQAS plays a complementary role to IQC. According to ISO 15189, the laboratory is encouraged to use a third party control material similar or identical to the patient sample matrix. followed. Daily QC values are documented and %CV calculated. Control charts are maintained to demonstrate the stability of the analytical measuring systems. Two EQAS programs are followed- ACBI/CMC EQAS and Randox International Quality Assessment Scheme (RIQAS). In the monthly ACBI/CMC EQAS program, the laboratory's performance is compared with other labs and assessed by a Variance Index Score. The three fortnightly RIQAS programs clinical chemistry, immunoassay and cardiac are interpreted by Target Score and Standard Deviation Index.

Conclusion: The quality indicators give credibility to the laboratory's performance, along with improvement in turnaround time. They also provide information on critical aspects of the Methods: We use three levels of QC material- two process, identify the necessity of improvement levels are run twice a day at appropriate intervals actions and improve overall patient safety. in different combinations and multi-QC rules are Abstract No: 6.27

Quality Indicators for Measuring the Process Performance of a Clinical Laboratory

Dr.R.Balambal Sundaram Medical Foundation, Dr.Rangarajan Memorial Hospital, Chennai. Objective: Quality Indicator is a specific measurement of the performance of processes used to find whether they are in control or to identify scope for improvement. It provides a way of measuring, monitoring and improving the quality of care and services. It is useful to reduce the laboratory errors and to improve the quality in medical laboratories. The entire laboratory processes - pre examination, examination and post-examination processes, need to be monitored with quality indicators. Methods: The indicators identified for each lab process are as follows: - Pre examination process: Number of - Requests without proper details, erroneous samples taken, samples rejected, TAT not achieved, repeat samples, needle stick injury reported, delayed transport of Urgent samples,

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repeated training given on specific area. Examination process: Number of - Breakdowns of equipments, kits unavailable, repeat tests, CA taken for IQC (Out of range situation, imprecision, systematic error), % Error > 10 in ILC, EQAS results >1 SD/Accuracy Score > 3. Post Examination process: Number of - Delayed reports, erroneous reports, complaints from physicians & patients. The Quality indicators can be converted into the objectives set in SMART format (Specific, Measurable, Achievable, Abstract No: 6.28

Realistic and Time bound). The target to be achieved for all the above should be `Nil'. Then the achieved percentage will be 100. The actions taken to achieve this 100% will help in the continual improvement of the Quality management system processes. Conclusion: Medical testing laboratory will have a continuous assurance of Customer satisfaction, for providing consistent, accurate and timely test results when the process performance is monitored with these Quality indicators.

Serum Protein Electrophoretic Pattern of Chronically ill Patients in a Tertiary Care Hospital

Vijayasree N Educare Institute of Dental Sciences, Malappuram, Pin 676 504 and George Abraham, Malabar Institute of Medical Sciences, Calicut, Pin 673 016; e mail ­ [email protected] Background: Protein electrophoresis is the technique employed in major clinical laboratories for screening abnormal proteins pattern in serum and other biological fluids. Serum protein electrophoresis is extensively employed for the diagnosis and prognosis of many chronic diseases such as myeloma, leukemia, nephrotic syndrome, autoimmune diseases and acute phase reactions. Objective: In the present study an attempt has been made to evaluate the changes, if any, in the serum protein electrophoretic pattern Methods: Patients with chronic diseases who were admitted in a tertiary care referral hospital (Malabar institute of Medical Sciences, Calicut) during the period from January to December 2006. Patients with any chronic diseases showing changes in their serum proteins pattern (as assessed by the estimation of serum total proteins by Biuret method and albumin by BCP dye binding method) were included in the study.

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During this period a total of eighty nine serum samples were selected and subjected for serum protein electrophoresis in Agarose gel (Ready to use Hydragel Protein K (20) kit supplied by M/S Sebia, France). Results: Eight type of abnormal electrophoretic patterns namely, hypergammaglobulinemia, inflammatory disease, hypergamma globulinemia with beta gamma bridge, monoclonal gammopathy with M band, polyclonal gammopathy , free light chain disease, hypogammaglobulinemia and bisalbumi-nemia were observed. Hypergammaglobulinemia and inflammatory disease were common constituting 48% and 27% of the total cases respectively. The changes in the electrophoretic pattern may be attributed to liver diseases or autoimmune diseases. In hypergammaglobulinemia and inflammatory diseases the females were predominantly affected (67% and 65% rrespectively). A seasonal variation

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in the prevalence of inflammatory disease was observed with maximum prevalence in summer. Monoclonal gammopathy with M band and hypergammaglobulinemia with beta gamma bridge were seen in 4 patients each whereas polyclonal gammopathy and freelight chain disease were seen only in 3 cases each. Bisalbuminemia and hypogamma-globulinemia were very rare and could be detected only in one case each.

Conclusion: From this study it can be concluded that improved techniques of electrophoresis along with software support is more useful in facilitating the diagnosis of various diseases related to acute phase reactions, chronic diseases and hyper/hypo immune reactions in addition to multiple myeloma.

Abstract No: 6.29

Immunocharacterisation of Polyclonal and Monoclonal Antibodies Raised against Cobra Venom Toxin

Lakshmi.A 1, Dr. K.S.Vasudevan2, Mr.A.Mathialagan 2 and Dr.S.Subramanian1 1Dept.of Biochemistry, University of Madras, 2 Dept.of Immunotoxicology, Mediclone Biotech Pvt. Ltd., CHENNAI. Objective: Snakebites continue to be a major concern in India. The polyvalent antivenom available possess more problem in anaphylactic shocks. The development of monoclonal antibodies specific for various toxins of the venom is the only solution. In the present study, an attempt was made to characterize the various toxic compounds of cobra venom and to raise specific monoclones against the characterized toxic compounds of cobra venom. Method: Cobra venom was characterized by Ionexchange chromatography and the protein levels in each fraction was estimated by Lowry's Method. The protein was purified by SDS and NATIVE PAGE. The hemolytic activity of the fractions was determined by direct microtitre plate and indirect blood agar plate methods. The Phospholipase A2 activity of the fractions was determined by the direct tube and the indirct plate methods. Cytotoxicity assay was performed to determine the cytotoxic activity of the fractions. Invivo immunization of balb/c mice was carried out after determination of the lethal dose for cobra venom. Hyperimmunized status of the mice was confirmed by Double Immuno Diffusion method. Cell fusion for hybridoma production was performed and the monoclones were tested for cross reactivity by ELISA technique. Results: Cross reactivity for cobra venom was checked using various monoclonal antibodies and Krait venom antibodies were also found to be reactive with the cobra venom antigen. Each fraction should be used for the production of monoclonal antibodies and further evaluation of these antibodies could be used for therapeutic purpose. This preliminary study could be used as basis for studies on Naja naja toxin and its neurotoxic nature. Also, this toxin can be used to treat painful neurological movement disorders.

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Abstract No: 6.30

Development of a Novel Method to Eliminate the Negative Interference of Bilirubin on Creatinine Assay by Jaffe's Kinetic Method in Icteric Sera.

M.M.Goyal and A.Basak Background: The negative interference of bilirubin in creatinine assay by Jaffe's kinetic method is well documented. To remove this interference various methods had been tried. So far, very few methods provide solution to this problem with limitations. Objective: To develop a simple and economic method to remove negative interference of bilirubin on creatinine estimation. Methods: Icteric sera were treated with reagent `X' followed by precipitation of proteins. After this, Creatinine was estimated by Jaffe's kinetic method in colourless supernatant. Results: Our findings showed that reagent `X' binds covalently bilirubin (both conjugated and unconjugated) with serum proteins which were precipitated out by precipitating reagent. Creatinine was estimated and accurate values were found out multiplying by a factor. Conclusion: Our devised method effectively removes bilirubin interference in creatinine estimation. It is a simple procedure with good repeatability and economic also.

Abstract No: 6.31

Clinical Biochemistry : The Goldmine of Challenges and How do we Face It

K. Sorte, K. Palandurkar, M.M. Goyal and A. Basak Background: Common Clinical Biochemistry investigations done round the clock often pose great challenges to the Biochemists. Simple example : in lipaemic serum no investigations can be done. Sometimes, the procedure is cumbersome, stability of reagents is a problem or no method is available to estimate biomolecules of clinical interest. Interferences of endogenous & exogenous molecules, specially different drugs administrated to the patients, make the situation more complicated. Cost of the investigations by commercial kits is high and many poor patients can't afford it. Results: In our Clinical Biochemistry Laboratory, we have developed few new methods to overcome different day to day problems faced. Our team has developed some simple methods to remove

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interferences of bilirubin in creatinine, cholesterol, triglyceride, uric acid estimation in icteric sera. A very simple method of porphyria diagnosis, detection of chyle in body fluids have been developed. A very short procedure (1½ min.) of plasma glucose estimation has been developed for detection and prompt initiation of therapy of hypoglycaemic patients as a life saving measure. Plasma ketone bodies estimation is on the way. A simple devise has been developed for collection of two drops of blood by finger prick for glucose estimation obviating venepuncture. These new methods will be discuss. Conclusion: Hence, future is awaiting for us with many avenues open to solve the problems in Clinical Biochemistry for better patients' care as well as to save our patients' lives.

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Abstract No: 6.32

Hyperhomocysteinemia and Folic Acid, Vitamin B12 Levels and their Correlation in Eclampsia

Shahid A. Mujawar1, Vinayak W. Patil1, Rekha G. Daver2 and Narayan A. Madrasi3 1 Department of Biochemistry, Grant Medical College and Sir J.J. Group of Government Hospitals, Byculla, Mumbai ­ 400008, Maharashtra, India, 2Department of obstetrics and Gynecology, Grant Medical College and Sir J.J. Group of Government Hospitals, Byculla, Mumbai ­ 400008, Maharashtra, India, 3Department of Biochemistry, Government Medical College and General Hospital, Miraj ­ 400008, Maharashtra, India. Email: [email protected] Objective: An elevated level of circulating total homocysteine (tHcy) is recognized as an independent risk factor for cardiovascular diseases. Moreover, determinants of hyperhomocysteinemia, such as low concentrations of folic acid and vitamin B12 coenzymes involved in tHcy metabolism are also associated with increased risk of vascular damage. Therefore, in the present study we estimate the levels of tHcy, folic acid, and vitamin B12 and their relationship in patients with Eclampsia. evaluated by solid phase, competitive chemiluminescent assay method. These parameters were determined by means of Immulite 1000 analyzer.

Results: The statistical analysis of study group compared with normal control group, showed significant (p<0.001) decreases in serum folic acid, vitamin B12 levels whereas increased concentration of tHcy in all 50 eclamptic subject indicate hyperhomocysteinemia. Inverse and significant correlations found between serum tHcy Methods: 50 pregnant women with eclampsia in and folic acid, and vitamin B12 whereas, a positive the age group of 18 to 30 years were studied over and significant correlation was observed between a period of January 2008 to June 2009. Serum tHcy folic acid and vitamin B12 in study group. concentration was measured by competitive chemiluminescent enzyme immunoassay method. Conclusion: The present study found a significant Serum folic acid concentration was investigated association between serum folic acid, vitamin B12 by boil competitive, liquid phase, ligand labeled, levels and their deficiency indicate cardiovascular in eclampsia patients with and protein binding chemiluminescent assay risk method. Serum vitamin B12 concentration was hyperhomocysteinemia. Abstract No: 7.1

The Association between Serum Alkaline Phosphatase and ABO Blood Group

E.Sruti1, Nalini Ganesan2 and Anbazhagan.M3 1. PG student, 2. Professor, 3. Chief Biochemist, Department of biochemistry, SRMC and RI, SRU, Porur, Chennai. Objective: Alkaline phosphates (ALP) is a hydrolase enzyme that catalyzes hydrolysis of organic phosphate at alkaline pH. This enzyme is present in practically all of the tissues of the body and occurs at particularly high levels in the intestinal epithelium, kidney tubules, bones, liver and placenta. Small amount of intestinal ALP may also be present, particularly in the sera of individuals of blood groups `B' or `O' (secretors). Our objective was to estimate the amount of ALP

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in different blood groups of healthy individuals, by using TULIP diagnostic kit. The statistical and to compare their levels. analysis was done using student's t-test. Methods: Serum levels of total ALP was detected in 150 healthy subjects with different blood groups in the age groups of 20 ­ 40 years. 2ml of random blood sample was collected from the staff and students of Ramachandra Medical Center after getting their informed consent. Serum total ALP was assayed by dimension RXL using SIEMENS kit. In this kit. p-nitrophenylphosphate was used as an artificial substrate and the results were expressed in U/L. Blood groups were determined Abstract No: 7.2 Results: The mean value of serum ALP was higher in `B' and `O' blood group individuals when compared to `A' and `AB' blood group individuals, though this did not reach statistical significance. Conclusion: The results of the present study indicate that there was no significant change in the ALP level of 150 healthy individuals with different blood groups.

Guidelines and Recommendation for Laboratory Analysis in the Diagnosis

Dr. Alok Parekh, Dr.Maulik Nayak, Dr.Rita Shah and Dr. D.A. Trivedi Content: Assessment of plasma glucose remains the only diagnostic standard for diabetes. Patients can monitor glycemic control by measuring their own plasma of blood glucose with meters & by laboratory analysis of glycated hemoglobin. The Methods: An expert committee outlined evidence- potential roles of noninvasive glucose monitoring, based recommendations for the use of laboratory genetic testing autoantibodies, microalbumin, analysis in diabetes. An external panel of experts proinsulin, C-peptide, and other analytes are reviewed guidelines & customized in response to addressed. reviewers' suggestions. A revised draft was presented at the AACC Annual Meeting in July, Summary: Based on published data or derived 2000. The recommendations were modified from expert consensus, the guidelines provide another time in response to oral and written specific recommendations. Several analytes are of remarks & further reviewed by the Professional minimal clinical value at the present time and Practice Committee of American Diabetes measurement of them is not recommended. Association. Background: The quality of scientific evidence supporting the use of several laboratory tests varies significantly in the diagnosis and management of patients with diabetes mellitus. Abstract No: 7.3

Establishing a Reference Range for Thiopurine Methyltransferase (TPMT) Activity in Indian Populations

Jayashree T1, Sadik A Mahammad1, Ramchandra Rao O1, Rupa Banerjee2and Nageshwar Reddy D2 1 Asian Healthcare Foundation, 2Asian Institute of Gastroenterology, 6-3-661, Somajiguda, Hyderabad-500083. Objective: Assaying the activity of thiopurine methyltransferase (TPMT), an enzyme metabolizing thiopurines used for treatment of

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various inflammatory conditions, is of clinical relevance for reducing the associated risk of acute toxicity. While reference ranges for TPMT activity

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have been established in other countries, limited information is available in the highly heterogeneous Indian population. The present study was thus conducted to establish a reference range for TPMT activity in Indian population.

Results: The histogram of combined population study showed a bimodal distribution of TPMT activity, with no subject having low TPMT activity (<3 units). The range of TPMT activity was wide (3 to 39 nmolh-1ml-1PRBC). The TPMT activity in the females was found to be significantly less than Methods: TPMT catalyses the conversion of 6- those of males. mercaptopurine (6-MP) to 6methylmercaptopurine (6-MMP) which can be Conclusion: This is the first comprehensive TPMT measured by HPLC. TPMT activity was assayed phenotype study from India for establishing in red cell hemolysates obtained from 200 healthy reference range for Indian population. The cut-off volunteers (168 males and 32 females) employing point for intermediate and low TPMT activity is HPLC- based assay with UV detection of the usually determined by receiver operating curve product at 290 nm. Units of enzyme activity were (ROC) analysis between genotypes and expressed as nmoles of 6-MMP formed per ml of phenotypes. Studies are ongoing with regard to finding genetic polymorphisms in the above packed RBCs (PRBCs) per hour. samples. Abstract No: 7.4

Biochemical Impact of Lumirubin in Neonatal Hyperbilirubinemia

PS Kamble, RK Padalkar, PD Zende and VR Pandhare PDVVPF's Medical College, Ahmednagar, Maharashtra, India; Email: [email protected] than exchange transfusion28 in achieving prolonged reduction of bilirubin levels in infants with nonhemolytic jaundice. It is important to recognize that the efficiency of phototherapy extends well beyond its ability because it detoxifies bilirubin instantaneously and continuously, regardless of the serum bilirubin level. Phototherapy is associated with an increased incidence of watery diarrhea, thus increased fecal water loss. Unconjugated bilirubin can induced intestinal secretions, Method: Various types of fluoroscent light have suggesting that diarrhea may be a consequence of been used for phototherapy. Broad-spectrum the high concentration of bilirubin within the daylight, cool white, blue, monochromatic, intestinal lumen. "special blue" and a quartz-halide white light with tungsten filament that has a significant output in Conclusion: The structural isomers are more blue spectrum. important than the configurational isomers in terms Results: In all groups, light treatment appeared of the ability of phototherapy to lower the to have its greatest impact in the first 24 to 48 concentration of serum bilirubin within normal hours of therapy. Phototherapy is more effective level.

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Objective: In neonates and premature infants the threat of hyperbilirubinemia is significant. The intra-molecular cyclization of bilirubin occurs in the presence of light to form a substance called lumirubin (structural isomer). Lumirubin is cleared from the serum is much more rapidly then photoisomer as its excretion is so rapid. It is probably quantitatively more important than the configurational isomer for the bilirubin lowering effect of phototherapy.

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Abstract No: 7.5

Comparison of Cord Blood Atherogenic Index in Males and Females

Kharb S and Smiti Nanda* Departments of Biochemistry and Obstetrics and Gynaecology* Pt BDS PGIMS, Rohtak, India; Email: [email protected] Objective: Pathological processes for cardiovascular disease (CVD) and its risk factors have been rooted in childhood. Changes in maternal lipid metabolism during gestation control availability of lipid metabolites to be fetus. Abnormal lipoprotein profiles in childhood persist into adult life and gender-related factors might influence lipid levels. Methods: The present study was planned in two hundred women: study group consisted of 100 pre eclamptic women and their newborns; and hundred healthy pregnant women and their newborns served as controls. Maternal venous blood sample and cord blood from newborn were collected and were analyzed for lipid profile (total cholesterol, triglycerides, HDL cholesterol, VLDL cholesterol, and LDL cholesterol), apolipoproteins: ApoA-I, ApoB. Atherogenic index (A.I.) was calculated as ratio of Apo-B to ApoA-I. Conclusion:The results in terms of maternal and cord blood atherogenic index and comparison between two groups and influence of genderrelated factors on lipid profile will be discussed.

Abstract No: 7.6

Quality Control in Clinical Chemistry: Minimal Requirements for an Acceptable Program

Dr. Manju Bala Pahwa* and Dr. Veena Singh Department of Biochemistry, Pt. B.D. Sharma, University of Health Sciences, Rohtak-124001, [India] Background: A properly functioning quality control program is one of the most effective safeguards available to ensure good quality of laboratory work, particularly in clinical chemistry. No longer is it sufficient to feel that laboratory results are satisfactory. It is rapidly becoming necessary to know that they are satisfactory and to have tangible evidence of competence. Quality control measures provide that knowledge and evidence. Objective: The main objective of quality control

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is to ensure good overall performance and thus greatly to enhance the confidence that can be accorded to any single estimation. Quality control programs stress accuracy at the bench level. Conclusion: This approach in itself is praiseworthy, for if the estimation is in error, the whole report is in error. Since each laboratory has its own problems, variations in quality control programs can be expected. Components of a minimal program of quality control in clinical chemistry will be discussed.

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Abstract No: 7.7

Quality Indicators: A Tool for Continuous Quality Improvement in Clinical Laboratories

S.Subramaniam and Shyama Subramaniam Department of Biochemistry, Apollo Hospitals, Chennai ­ 600 006. Background: Quality indicators (QI) are defined as specially identified measurable elements for a laboratory performance giving a base for evalution and improvement of the laboratory quality. control results, external quality assurance results, imprecision, inaccuracy, total error and for postanalytical phase number of tests not completed, reports with wrong patient data, average time for critical results, reporting number of critical results successfully reported, reports exceeding TAT, customer satisfaction, number of reports corrected or withdrawn, LIS downtime episodes, technical errors, etc.

Methods:The QI should measure the laboratory performance in the key function related to patient care and satisfaction and are usually selected as parts of the total testing process such as preanalytical, analytical and postanalytical phase Conclusion:The establishment of indicators may of laboratory activities. allow us to detect and quantify problems in the Results: QI for preanalytical phase may include clinical laboratory. Follow up of these indicators error in patient identification, erroneous request, will make it possible to determine whether test order appropriateness, inadequate sample improvement measures results in fewer error and missing sample, needle stick injuries, etc. For higher quality. analytical phase, QI may include internal quality Abstract No: 7.8

Serum Sodium and Potassium Level in Emesis Gravidarum

Sana Hasan*, Dr. B.K.Agarwal* and Dr. Neelkamal Kapoor** *Dept. of Medical Biochemistry **Dept. of Pathology, Gandhi Medical College, Bhopal. Email:[email protected] , [email protected] Background: Emesis Gravidarum , or pregnancy sickness is a condition effecting more than half of all pregnant women. Extreme cases includes dehydration, weight loss, acidosis, hypokalemia and hyponatremia. Methods:The study includes 30 controls of normal pregnancy in first trimester and 30 cases of Emesis Gravidarum also in first trimester among the age group 18-30 years of age. 2 ml of blood is withdrawn under aseptic precautions to determine Serum Sodium and Potassium level by Carelyte Electrolyte Analyzer. Results: Highly significant decrease in K+ level (p<0.001) was found with significant decrease in Na+ level (p<0.01) as compared to control group where Na+ and K+ level were found to be in reference range. Conclusion: The present study concluded that Emesis Gravidarum is a significant problem in nearly 60% of all pregnant women. Low levels of K+ and Na+ is associated with Emesis Gravidarum in pregnancy.

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Abstract No: 7.9

Study of Serum Fluoride and Sialic Acid Levels in Osteosarcoma

Sandhu R, Lal H, Kundu ZS* and Sandeep, Kharb S Department of Biochemistry and Orthopaedics*, Pt B D Sharma University of Health sciences; Email: [email protected] Background: Osteosarcoma is a rare malignant bone tumor most commonly occurring in children and young adults presenting with painful swelling. Status of fluoride levels in serum of osteosarcoma is still not clear. Recent reports have indicated that there is a link between fluoride exposure and osteosarcoma. Glycoproteins and glycosaminoglycans are integral part of bone and prolonged exposure to fluoride for long duration has been shown to cause degradation of collagen and ground substance in bones Methods: The present study was planned to analyse serum fluoride, sialic acid, calcium, phosphorus, and alkaline phosphatase levels in twenty five patients of osteosarcoma and age- and sex-matched subjects with bone forming tumours other than osteosarcoma and musculo-skeletal pain (controls, 25 each). Abstract No: 7.10 Results:Mean serum fluoride concen-tration was found to be significantly higher in patients with osteosarcoma as compared to the other two groups. The mean value of fluoride in patients with other bone forming tumours were though approximately 50% of the group of osteosarcoma, however was significantly higher when compared with patients of group I. Serum sialic acid concentration was found to be significantly raised in patients with osteosarcoma as well as in the group with other bone forming tumours as compared to the group of controls. Conclusion: There was however, no significant difference in the group of patients of osteosarcoma when compared with group of patients with other bone forming tumours. These results showing higher level of fluoride, sialic acid and calcium in patients with osteosarcoma compared to others suggesting a role of fluoride in the disease.

Prevalence of Diabetes Mellitus in Urban Region of Chennai City- a part of PurseHIS study

Satyajeet Giri1, S.Thanikachalam2 and K.Sowmya3 1.Senior Research fellow 2. Professor, Department of Cardiology, 3. Assistant Professor, Department of Biochemistry, SRMC and RI, SRU, Porur, Chennai Email: [email protected] or [email protected] Objectives: Endovascular disease is more common in the south Asian region, especially in India. Cardiovascular disease is the leading cause of death in India and its contribution to mortality is rising. Hence, the concept of the study is to identify the various risk factors for Endovascular disease affecting Cardiovascular, Peripheral vascular and Cerebro vascular system.

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Methods: A total of 2226 subjects from the urban Chennai were enrolled for this study. Clinical history, foods habits, physical activity and socioeconomic status were collected. Excluding known diabetics all other subjects underwent OGTT and were categorized as normal glucose tolerance, newly diagnosed diabetics and pre diabetics based on ADA guidelines. HbA1C was also estimated

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for all the subjects and values upto 6 % was HbA1C is 14.3%, 87.8%, and 42.6% in NGT, considered to be normal. Diabetics and Pre diabetics respectively. Results: Out of 2226 subjects, 1275 were found to have normal glucose tolerance (NGT), 392 were already diabetic (18%), 165 were newly diagnosed diabetic (7%), 191 and 203 were respectively IFG and IGT (9%). The percentage of abnormality in Abstract No: 7.11 Conclusion: The prevalence of Diabetes Mellitus in the selected population is alarming (25%). Hence urgent attention is required for its prevention.

A Study of Serum Ferritin Levels in Type -2 Diabetic Patients

Shiv Ram Krishn and Ekta Agarwal . Email Id: [email protected] Objective: The objective of the study is to compare serum ferritin levels of diabetic patients with the control group and to study the effect of body iron stores on diabetic complications. concentration was 104.0 ng/ml in patients suffering from diabetes. A directly proportional relationship was found between serum ferritin and blood sugar levels. The correlation was found to be significant.(P=0.006). Diabetics with increased serum ferritin levels have significantly poor glycemic control which was reflected by higher levels of GHb. Similarly, significant relationship is evident between increased serum ferritn levels and neuropathy, nephropathy, retinopathy and hypertension ,whereas no significant relationship was found between increased serum ferritn levels and Ischaemic Heart disease(P=NS).

Methods: The study was done on 50 obese type2 diabetic patients. They were compared with an equal number of age, sex, BMI matched controls, selected randomly to find out the correlation between serum ferritn and blood sugar levels. The patients were tested for the following parameters: Serum Ferritin, Fasting Blood sugar, postprandial blood sugar, random blood sugar, lipid profile & GHb using standard methods. Patients were also evaluated for Diabetic Retinopathy, Nephropathy Conclusion: It can be interpreted from the study that elevated iron stores reflected in elevated serum , Coronary Artery disease and Neuropathy. ferritin, may induce baseline metabolic Results: Mean ferritin concentration was 70.0 ng/ abnormalities resulting in diabetes mellitus. ml in the control group,whereas mean ferritin Abstract No: 7.12

Comparison of Serum and Plasma Uric Acid

Dr. Shivraj Gowda, Dr. P.B. Desai, Shruti Kulkarni, Sonal Vernekar, Avinash A.K.Math, Vinayak Hull and Mahantesh Ilkal Department of Biochemistry, J. N. Medical College. Belgaum ­ 590010. Email- shru_kul @yahoo.co.in Background: Uric acid is an organic compound of carbon, nitrogen, oxygen and hydrogen with the formula C 5H4N4O 3. In human blood, the normal uric acid concentration is 3.6 mg/dL to 8.3 mg/dL. An attempt was made to find out the difference in the values between serum and plasma in humans by using phosphotungstate method of Henry et al. It is a colorimetric method based on the reduction of phosphotungstate by urate in

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carbonate buffer at pH 10.2. It is non-specific but Conclusion: Review of literature have shown that widely used method in all laboratories. some laboratories are switching to plasma because serum specimens have several inherent problems: Methods: A pilot study was carried out taking 15 (a) an increase in turnaround time because of the serum samples sent to biochemistry laboratory and time necessary for clot formation, especially in same individual plasma was collected from patients receiving anticoagulant therapy; and (b) pathology laboratory of our hospital and uric acid the risk of fibrin clot interference on automated was estimated by phosphotungstate method of analyzers, especially those with a common sample Henry et al. probe and no clot detection ability. Hence an Results: There was negligible difference between attempt was made to see the difference using conventional method. plasma and serum uric acid levels. Abstract No: 7.13

Tourniquet: Lacunae of Preanalytics

Smita S. Sonoli, and Anuradha B. Patil Department of Biochemistry, J.N. Medical College, Belgaum-590010 Objective: This study was aimed to investigate the influence of prolonged stasis, created during blood sample collection, on five common biochemical parameters. Method: Five blood samples were collected by venepuncture from 20 healthy individuals. One before applying standard pressure and four after applying standardized pressure of 60 mm of Hg(1 and 3 minutes) and 90 mm of Hg (1 and 3 minutes) with help of sphygmomanometer. Glucose, Total protein, Albumin, Urea, Creatinine were estimated by kit method and analysed on semiautoanalyzer manufactured by Transasia in collaboration with Erba diagnostics (Germany). Abstract No: 7.14 Results: With increase in pressure and time there was increase in the levels of total protein and albumin and decrease in levels of glucose and urea (p <0.05). However creatinine did not show change in values on short term stasis, but prolonged stasis (90 mm of Hg for 3 minutes) showed decrease in values (p < 0.02). Conclusion: From our results, we conclude that the parameters in question are influenced by the duration and magnitude of pressure applied, hence tourniquet should be used meticulously. Phlebotomists should be educated regarding the usage of tourniquet so that we can identify and prevent laboratory errors.

Simplified Procedure for Cholesterol Determination

Dr. Shivraj Gowda, Dr. P.B. Desai, Sonal N. Vernekar, Vinayak Hull, Avinash A.K.Math, Shruti Kulkarni and Mahantesh Ilkal Department of Biochemistry, J. N. Medical College. Belgaum ­ 590010, Email: [email protected] Background: Cholesterol is a sterol that is found in all animal tissues and serves many important

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physiological functions. High blood cholesterol has been associated with heart disease. Hence total

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cholesterol estimation is used to diagnose and monitor the disease. Liebermann described the colour reaction of sulphuric acid with the solution of cholesterol in acetic anhydride. Burchard reported that more intense blue-green colour is produced when acetic acid and sulphuric acid are to be added to the solution of cholesterol in chloroform. Since time Liebermann-Burchard reaction has been widely used in colorimeteric reaction for estimation of cholesterol in biological fluids. By using the same principle, an attempt was made to simplify the procedure.

sodium sulphate and Reagent B without anhydrous sodium sulphate. A Cholesterol standard graph was prepared for both reagents and test samples were extrapolated and compared with Autoanalyzer. Results: In case of Reagent A, turbidity was observed and took 20 minutes to attain clear mixture. In case of Reagent B, turbidity was cleared off within 10 minutes and remained stable till 40 minutes.

Conclusion: The above method for preparation of Reagent B provides a simple procedure in reducing Methods: Modified reagents were prepared along the turbidity and also maintaining the colour with routine reagents. Reagent A with anhydrous stability.

Abstract No: 7.15

Study of Lipid Profile and Glycosylated Hemoglobin (HbA1 C) in Patients of Diabetes Mellitus

Tripti Saxena*, B.K. Agrawal*, and V.K. Sharma,** *Dept. of Biochemistry, **Dept. of Medicine, Gandhi Medical College Bhopal. [email protected] Background: Diabetes mellitus is chronic metabolic disorder, with an absolute or relative insulin deficiency, fasting hyperglycemia, glycosuria leading to development of atherosclerosis, microangiopathy, nephropathy and neuropathy. It is increasingly believed that strict glycemic control in diabetes prevents the progress of late vascular or neurological complications, therefore proper assessment of glycaemic control becomes an essential part of monitoring. MethodsPresent study comprised 100 patients of DM ,.(including both type 1 and 2). Fasting blood sample were taking and following investigations were done:Blood glucose. Total CHO CHOD-PAD method by Allainet et al. TG GPO-POD method by Werner method. HDL by Burnstein et al. LDL by Friedwald's and Fredrickson's formula. Glcosylated hemoglobin by cation exchange resin method. Results Patients have been devided in to good, fare and poor control groups on the basis of glycoselated hemoglobin (group 1 HbA1 C-6-8 gm%, group 2 HbA1 C 8-10%, group 3 HbA1 C>10gm% ) and found that total cholesterol content(>200mg%), LDL(>100mg%), Triglycerids(>150mg%) were significantly high in group 3(p<0.05) in comparison to group 1 and 2.

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Abstract No: 7.16

Use of Tris-Borate Buffer in Agarose Gel Electrophoresis of Serum Proteins

Mahesh S1, Naureen Anwar1, Yeshoda K1, Saifa Yasmin1 and Dr. Nalini K1 1 Department of Biochemistry, Kasturba Medical College, Manipal, Karnataka-576104; E-mail for correspondence: [email protected] Objective: Agarose gel electrophoresis of serum proteins is a simple technique which provides wealth of information in diagnosis of many clinical conditions. Barbitone buffer has traditionally been used in this technique and has been shown to give proper separation of components. But difficulty in access to barbiturates has necessitated the need of an alternative buffer. Some previous works have used tris-borate in SDS PAGE, moving boundary and immunoelectrophoresis of serum proteins. The present work is carried out to optimise trisborate buffer as an alternative to barbitone buffer for use in agarose gel electrophoresis of serum proteins. Methods: Tris-borate buffer of strength 0.05M and pH 8.6 is used for the electrophoretic Abstract No: 7.17 procedure. Electrophoresis is run for 21/2 hours at a current of 3mA/slide. The bands are visualized by staining with amido schwatz. Patterns in various disorders are then compared with the patterns obtained with barbitone buffer. Results: Comparison of the patterns obtained by the use of tris-borate and barbitone buffer is presented. Similar numbers of electrophoretically distinct components are found by the use of either buffer. Mobilities of these components are also comparable. Conclusion: The similarity in the electrophoretic pattern and mobility of the components with that of the classical pattern indicates that tris-borate can be a suitable alternative for barbitone buffer.

Use of the Laboratory in Prediction of Outcome in the High-Risk Newborn

Dr. Alok Parekh, Dr.Maulik Nayak, Dr.Rita Shah and Dr. D.A. Trivedi This paper assesses ability to envisage survival & neurodevelopment outcome in the newborn period. The laboratory plays a principal role in diagnosing some of these. For instance, prenatal & new born screenings are important in the diagnosis of chromosomal anomalies and inborn error of metabolism. Abnormal bilirubin, glucose and pH values in the newborn periods are risk factors for death & abnormal neurodevelopment, & the degree of abnormality conveys additional information. Conventionally, prognosis based on individual risk

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factors or disease states. Many newborns have multisystem disorders, & it is only when multiple variable are considered that result can be predicted. Three neonatal scores that integrate multiple variables are discussed. Methodologic intricacy in determining outcome are reviewed and illustrated with survival and morbidity rates of very premature babies. The laboratory is one of many prognostic variables. The appraisal of laboratory services are provided is difficult but important.

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Abstract No: 7.18

Standardization of DAM Method by Single Step

Dr. Shivraj Gowda, Dr. P.B. Desai, Vinayak Hull, Avinash A.K.Math, Shruti Kulkarni, Sonal Vernekar and Mahantesh Ilkal. Department of Biochemistry, J. N. Medical College. Belgaum ­ 590010. Email: [email protected] Background: Most of the laboratories use urea nesslerisation method which was found to be tedious involving deproteinisation and incubation for 20 minutes. The estimation of urea by diacetyl monoxime (DAM) method is very simple and quick technique of analysis. The reaction between urea and DAM was established by Fearon in 1935. Literature have shown many modifications of this method, by changing the acid concentration, oxidant ions and use of 4-amino antipyrine, glucuronate, glucuronate glucosacchric acid, thiosemicarbazide (TSC) to stabilize the colour. The reaction occurs between urea and DAM in presence of strong acid reagent, ferric ion and thiosemicarbazide (TSC) to give a pink colour and can be measured colorimetrically. Keeping above principle, an attempt was made to standardize DAM method in single step. Methods: Mixed acid reagent contains Stock acid reagent, Mixed colour reagent ( Stock colour reagent A + Stock colour reagent B), Stock urea standard and Working urea standard were prepared. All the stock reagents are stored in brown bottle at room temperature and are stable for 6 months. The colour reagent is prepared fresh at the time of analysis by mixing distilled water, mixed acid reagent and mixed colour reagent in the ratio1:1:1. This is single step method. The working standards were taken in respective test tubes, volume was made to 1ml with distilled water, 3ml of colour reagent was added, tubes were mixed well and kept in boiling water bath for 15 minutes, the tubes were cooled and the pink colour developed was measured at 540nm. Result: A linear graph has been obtained. Conclusion: DAM method in single step has been standardized

Abstract No: 7.19

Effect of Feeding Practices in Neonatal Hyperbilirubinemia Subjects

PD Zende, RK Padalkar, PS Kamble and VR Pandhare PDVVPF's Medical College, Ahmednagar, Maharashtra, India; Email: [email protected] Objective: Jaundice is most common vexing problem in new born period. Especially feeding practice in premature neonates in certain mother shows long term adverse effect of hyperbilirubinemia. Hence present study was undertaken to identify the long term adverse effect of hyperbilirubinemia in premature neonates, at different age group (37 to 40 weeks) and birth weight (0.9 to 3.0 kg) during onset of jaundice. Methods: A total 70 newborns of the age group 1 to 8 days were selected for study. 35 infants were on breast feeding and rests 35 were on bottle feeding. The serum Total and Direct Bilirubin

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parameter studied under above heading and is carried out by Malloy and Evelyn (1937) method. Results: There was slight decrease in mean value of total and indirect bilirubin with increase in gestational age and birth weigh of neonates during onset of jaundice and mean value of total and indirect bilirubin found significantly increased in breast fed neonates of certain mother compared to bottle fed neonates. Abstract No: 7.20

Conclusion: The present study has identified how certain mothers having 3- á, 20-â- pregnanediol factor in their milk causes hyperbilirubinemia in neonates and it is suggested that breast feeding is to be encourage only in the absence of aggravating factors such as high milk levels of the steroids and nonesterified fatty acids.

Novel Therapeutic Properties of Nano Silver*

Siddhartha Shrivastava, Sunil Singh and Debabrata Dash Department of Biochemistry, Institute of Medical Sciences, Banaras Hindu University, Varanasi-221005 Background: Nanotechnology has been finding increasing applications in biology and medicine. Nano silver has already been known to have antibacterial property. We recently have synthesized highly stable biocompatible nanoparticles of silver (10-15 nm diameter) with enhanced antibacterial effect against gram negative organisms 1 . Nano silver equally prevented growth of multi drug-resistant bacterial, which can have useful clinical application. We subsequently discovered that nanoparticles of silver, and not of gold, have innate anti-platelet property2. They effectively prevent integrin IIb3mediated platelet responses (aggregation, adhesion to immobilized matrix, secretion of granule contents, intracellular calcium and F-actin fluxes, flow cytometric evaluation of surface epitopes and changes in tyrosine phosphoproteome) in a concentration-dependent manner. Methods: When nanoparticles were intravenously injected into different mice strains, there was dosespecific inhibition of platelet aggregation, without any change in tail bleeding. Electron microscopy

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shows that nano silver accumulates within platelet granules and reduces inter-platelet proximity. Results: Our findings further suggest that these nanoparticles do not confer any lytic effect on platelets and thus hold immense potential to be promoted as anti-platelet / anti-thrombotic agent. Equipped with dual properties (antibacterial and antiplatelet), nano silver has unique application potentials in situations like coronary stents, where it can prevent bacterial infection while keeping platelets in inhibited state. References: 1Shrivastava, S., Bera, T., Roy, A., Singh, G., Ramachandrarao, P. & Dash, D. (2007) Nanotechnology, 18: 225103 (Characterization of enhanced antibacterial effects of novel silver nanoparticles) Shrivastava, S., Bera, T., Singh, S., Singh, G., Ramachandrarao, P. & Dash, D. (2009) ACS Nano, 3: 1357-1364 (Characterization of anti-platelet properties of silver nanoparticles) Acknowledgment: Research funded by Department of Biotechnology, Govt. of India, and Alexander Humboldt Foundation, Germany

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Abstract No: 7.21

Evaluation of Procalcitonin as a Diagnostic Marker of Sepsis

Ms. Nandini C and Dr.N.Leela Menon Dept of Biochemistry, Amrita School of Medicine, Kochi, Kerala, INDIA. Email: [email protected] Background: Sepsis is a systemic response to the curve. P value < 0.05 was considered to be infection, which causes organ failure and death in significant. severe cases. Results: The sensitivity of Procalcitonin, C Objective: To evaluate the reliability of reactive protein, WBC count, Differential procalcitonin as diagnostic marker of sepsis by leukocyte counts and culture results are 80%, studying it's sensitivity and specificity and 76%, 63%, 30%, 35% respectively. The specificity comparing it with that CRP, WBC counts, DLC of Procalcitonin, C reactive protein, WBC count and microbial culturing. Differential leukocyte count and culture results are 55%, 30%, 55%, 63%, 85% respectively. The Methods: The study was carried out at Amrita median plasma PCT and CRP concentration in Institute of Medical science and Research centre, septic and non-septic group was 1.34, 0.39 and Kochi. 71 blood samples were collected from the 23.4, 26.9 respectively. P value > 0.05, so there is Clinical Biochemistry lab. Procalcitonin in the no statistically significant difference between PCT samples were estimated using and CRP. The area under the ROC curve was 0.639 Immunoluminometric assay using a luminometer for PCT (95% CI, 0.488 - 0.744) compared with (LUMAT LB 9507) and C-reactive proteins 0.556 for CRP (95% CI, 0.498 ­ 0.695) .p value > estimated using Immunoturbidimetry (OLYMPUS 0.05, there was no statistically significant AU 2700). Data regarding the WBC count, difference between the sensitivity and specificity Differential leukocyte count and microbial of PCT and CRP. cultures results were obtained from Hematology and microbiology department respectively. The Conclusion: This study conclude that sensitivity parameters were compared to find out the most and specificity of Procalcitonin in the diagnosis reliable diagnostic marker of sepsis. The study also of sepsis similar to that of CRP but far better when assessed PCT and CRP as markers of sepsis by compared toWBC count, Differential leukocyte comparing their plasma concentrations in septic count and microbial culturing. But since and non-septic group using Mann ­Whitney test. Procalcitonin is an early marker, a combination of The two markers were compared using Receiver all parameters would provide better diagnosis Operative Curve (ROC) the respective areas under Abstract No: 7.22

Comparison of Various Parameters with Serum Free Light Chain Assay in the Diagnosis of Multiple Myeloma

Mrudula EV and Dr Sajitha Krishnan Department of Biochemistry, Amrita Institute of Medical Sciences and Research Center, Kochi; Email ID: [email protected] Background: Multiple myeloma is the second most prevalent blood cancer after Non-Hodgkin's

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lymphoma. It is characterized by excessive numbers of abnormal plasma cells in the bone marrow and overproduction of intact monoclonal immunoglobulins or Bence-Jones protein (free monoclonal ê and ë light chains). Objective: To compare the efficiency of serum free light chain (sFlc) assay with other biochemical parameters in the diagnosis and prognosis of multiple myeloma. Method: 56 patients diagnosed with Multiple Myeloma at Amrita Institute of Medical Sciences were included in the study. They were grouped into two - those with intact immunoglobulin multiple myeloma (IIMM) and those with light chain multiple myeloma (LCMM). Biochemical parameters such as total protein, albumin, globulin, creatinine were determined by photometric method in OLYMPUS AU2700, IgG, IgA, IgM, beta-2 microglobulin, kappa & lambda were Abstract No: 7.23

determined by immunoturbidimetric method in OLYMPUS AU400. Serum protein electrophoresis of all the patients was done by gel electrophoresis. Results: IIMM constituted about 56% and LCMM about 34% of the total MM patients. Among these patients IgG had the highest incidence. In patients with IIMM sFlc estimation was found to be 78% effective and in patients with LCMM sFlc estimation was found to be 95% effective compared to other parameters. Conclusion : sFlc can be used as a good diagnostic test and can effectively replace urine test for bence jones protein. It can also be used to monitor patients who cannot be done by electrophoresis and also help in rapid assessment of treatment responses. Thus serum free light chain assay along with other important tests can help in more efficient diagnosis and disease management in the case of multiple myeloma.

Importance of Tacrolimus Measurement and Assessment of its Toxicity in Monitoring Post Liver Transplant Patients

Anuja.P.S, Dr. K.N. Subhakumari and Dr. Jagathlal.P.C Department of Biochemistry, Amrita Institute of Medical Sciences and Research Center, Kochi. Background: Over the past 40 years, organ transplantation has become a rapidly expanding and important surgical specialty. After liver transplantation, the main problems encountered by the patients are rejection of the transplanted organ and opportunistic infection. So immunosuppressive drugs are given to post transplant patients and Tacrolimus is such a drug which is a powerful immunosuppressant. Side effects of Tacrolimus can occur and most of these effects are dose related. In case of Tacrolimus dose adjustment and change in immunosuppressive therapy can be done .So estimation of tacrolimus will be helpful in adjusting the dose. Thus a sufficient dose of tacrolimus can be given to patients to prevent rejection and to minimize the side effects.

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Objective: Present study was carried out to show the importance of tacrolimus assay in monitoring post liver transplant cases and to assess the toxicity due to Tacrolimus. Objectives of the study include quantitative estimation of Tacrolimus concentration in whole blood sample, estimation of total protein, albumin, globulin, bilirubin, ALP, ALT, AST, Creatinine, urea, uric acid, glucose, sodium, potassium and to assess the toxicity using the above parameters. Methods: 20 liver transplanted patients below 50 years of age were selected from Amrita Institute of Medical Science. Six month follow up was done for each patient. The blood sample was assayed for Tacrolimus by IMx analyzer using MEIA method. The liver function tests and renal function

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tests of each patient were also noted. Patients were grouped according to their Tacrolimus concentration. Group I (0-7 ng/ml) and Group II (7-15 ng/ml). Then the patient groups were compared with respect to their renal function and liver function tests. Results: A lower incidence of rejection was observed. In 20 transplanted patients the incidence of rejection was 20%. In this highest rate of rejection was seen in group II patients with a tacrolimus concentration 7-15 ng/ml. This study Abstract No: 7.24

also found nephrotoxicity and neurotoxicity in patients even within the therapeutic range, i.e. in group II patients. Results were compared using Mann-Whitney test with the help of SPSS software. Conclusion: In this study, toxicity was seen even within therapeutic range. So it is necessary to determine the concentration of Tacrolimus in the blood in order to establish a dosing regimen, maintain therapeutic levels within safe level. So Tacrolimus assay has to be done throughout the period of Tacrolimus intake i.e., through out the lifetime.

Six Sigma Analysis of Performance of Routine Chemistries at a Hospital Lab

Rashmi* and Dr.Narayani** *Technologist ** Head of Lab sciences, Wockhardt Hospital and Research centre, Mumbai Six sigma analysis of performance of chemistry assays in a lab helps to assess the performance on a common platform with other process parameters in a lab. obtained from analyzer data for each analyte. Sigma analsis for each of 20 chemistries was analysed using two levels of quality controls since it is difficult to obtain samples with values at decision limits for each analyte. We present and Aim:Objective of the study was to evaluate discuss the data for 20 analytes run on VITROS performance of routine chemistry assays in terms 5,1 FS chemistry analyzer. of sigma levels calculated considering imprecision and bias. This helps to judge quality of Conclusion: The sigma analysis showed that performance the analyser and competence of the around 50% of analytes on the analyzer performed operator in achieving accuracy and precision. at five sigma or more for both levels of controls used and 75% of analytes at or more than 5 sigma Method:The Westgard approach was utilized for when either level of control was considered. 35 calculating sigma levels i.e. Total error ­ Bias/ % of analytes demonstrated target six sigma level imprecision (CV%). Total allowable error(TE) of performance (>6 sigma) for the two levels of includes imprecision and inaccuracy and was controls used. Corrective actions have been obtained from CLIA limits. Bias is calculated from identified for the other chemistries performing end of cycle report on EQAS and imprecision below the five sigma level for improvement. Abstract No: 8.1

Hepatitis B Virus Sequence Database for Indian Isolates and Mutation Profiling Tool

Amit Tuteja1, Kaushal Madan3, Subraya K. Acharya2, Dr. K,Narayanasamy1..and Dr Swati Subodh1 1. The Centre for Genomic Applications, New Delhi, 2. All India Institute of Medical Sciences, New Delhi 3. Institute of Liver and Billary Sciences, New Delhi. Email Address: [email protected] Background: More than 350 million people in the world are chronically infected with Hepatitis

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B virus infected population. The database contains clinical, epidemiological, genotype and mutational data of patients. It would also act as a Genomic Sequence Repository for Indian isolates. Till date database has data from 472 samples which includes 236 baseline samples as well as the follow up data from patients who are on treatment. The database also contains information of previously known as well as novel mutations that have been found on sequence analysis of clinical isolates. Few of the mutations were also found to be associated with the drugs that patients were administered as a part of the treatment. At the end of the project the Methods: As a part of an ongoing Government database would be made web accessible to public funded project we have developed a HBV database and would also contain tools for sequence analysis for Indian isolates which aims to be a unique and mutation detection. resource for scientists working on HBV genetics, Conclusion:Once developed fully the Indian HBV evolution, variability, and vaccine and drug design Database will be useful as an extensive library of targeted to Indian population. HBV sequences well annotated with clinical and B Virus (HBV), and approximately 500,000 to 10, 00,000 deaths worldwide are attributed to it. India alone has world's second largest pool of carriers with a prevalence rate of ~2- 4% (approx. 36 million carriers) of which 10% are highly infectious. Wide variations in social, economic and health factors in different regions may explain variations in carrier rates from one part of the country to another. HBV is reported to be responsible for 70% of cases of chronic hepatitis and 80% of cases of Cirrhosis of the liver. Results: At present the database contains annotated data mostly from North Indian hepatitis Abstract No: 8.2 epidemiological data.

Association of Antioxidant Enzyme Gene Polymorphism in ATT Induced Hepatotoxicity in North Indians

S.V. Rana, Sanjeev K. Sharma, R.P. Ola, S.K. Sinha, J. Kaur, and K. Singh. Department of Super Specialty Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh. 160012, India Introduction: Drug induced hepatotoxicity in general, is involving toxic metabolite generation and its detoxification in the liver. Glutathione (GSH) plays an important protective role as intracellular free radical toxic reactive metabolites that are generated from biotransformation of drugs and xenobiotics. It's plays an important role in preventing ATT-induced hepatotoxicity. Deficiency in GST activity, because of homozygous null mutations at GSTM1 and GSTT1 genes may susceptibly to ATT-induced hepatotoxicity.

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Aim: Therefore, the multiplex PCR was used to determine the prevalence of deleted/addited genotypes of both GSTM1 and GSTT1 genes in ATT-induced hepatotoxicity patients. Methods: In the present study, a total of 100 north Indian adult patients of tuberculosis on ATT were enrolled. DNA used for polymorphic analysis was isolated from lymphocytes. The genetic polymorphic analysis for GSTM1 and GSTT1 genes were using multiplex PCR method with âglobulin gene as positive control. The presence or

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absence of GSTT1 (459 base pairs) and GSTM1 (215 base pairs) amplicons was determined in the presence of the control â-globulin gene (268 base pairs).

hepatotoxicity patients, 5 (31.3%) showed GSTM1, 4 out of 16 (25.0%) GSTT1 and 7 out of 16 (43.7%) GSTT1/GSTM1/â-globulin. While13 out of 84 (15.5%) showed GSTM1, 28 out of 84 (33.3%) GSTT1, and 43 out 84 (51.2%) GSTT1/ Results: Mean age of 100 patients was 45.6 years M1/ â-globulin in non-hepatotoxicity patients. (range 18-70 years). Out of 100 patients, 16 (16%) were ATT-induced hepatotoxicity and 84 (84%) Conclusion: This study showed GSTM1 was were of non-hepatotoxicity. Out of 16 significantly higher in hepatotoxicity patients as compared to non-hepatotoxicity patients. Abstract No: 8.3

Circulating Plasma DNA as a Diagnostic Tool ­ A Review

K.Ramadevi Institute of Biochemistry, Madras Medical College, Chennai. E mail: [email protected] Background: Free, extra-cellular nucleic acid are Method: Circulating fetal DNA molecules have found circulating in the plasma or serum. The been detected in maternal plasma from the first nucleic acid can be RNA, mitochondrial DNA or trimester onwards. This approach has been used genomic DNA. The existence of extra cellular for the prenatal investigation of sex-linked nucleic acids in the circulation was first reported diseases, fetal RhD status, and prenatal exclusion by Mandel and Metais in 1948. However, it was of ß-thalassemia major. only 3 decades later that further developments in Conclusion:Analysis of circulating nucleic acids this field have been reported, when it was in plasma is a promising noninvasive diagnostic demonstrated by Leon et al that cancer patients tool, requiring only a limited blood sample. The had much higher circulating DNA concentrations than those suffering from non-malignant diseases. use of plasma DNA and RNA has opened up opportunities to establish noninvasive tests for While only small levels are detectable in health, detecting cancer, as tumor markers, as a predictor higher levels are observed in disease, such as malignancy and sepsis. Similarly the discovery of mortality and sepsis, as a prognostic marker in of cell-free fetal DNA in maternal plasma in 1997 critically ill patients in addition to their use in has opened up new possibilities for noninvasive prenatal detection. prenatal diagnosis. Abstract No: 8.4

Polymorphism of p53 Gene Codon 72 in Indian Hyperlipedimic Patients with Cardiovascular Disease

K Ezhilarasan*, K Dhananjayan*, K.Anbarasu, A Sampath kumar**, A K Munirajan***, Shyama Subramaniam* and S. Subramaniam* *Department of Biochemistry, Apollo Hospitals, Chennai, **Department of Genetic Engineering, SRM University, Chennai, ***Department of Genetics, University of Madras, Taramani, Chennai. Objective: To study the prevalence of p53 codon Introduction: Atherosclerosis is a complex 72 polymorphism in Indian hyper lipedimic multifactorial disease involving multiple genetic patients with cardio vascular disease. and environmental factors. Recent studies

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implicate the role of P53 in atherosclerotic plaque stability by repressing vascular smooth muscle cell proliferation signals. Key polymorphism in codon 72 of p53 contribute to the alteration of its transactivating domain stability. Method: Blood samples were collected from the cardio vascular disease patients and used for lipid profile analysis. Hyper lipedimic patients, age and sex matched control patients were selected for the p53 codon 72 polymorphism study. Results: Analysis of CAD patients revealed an significant association between the disease and the Abstract No: 8.5

existence of diabetes (P=0.001). Also, the CAD patients had significantly higher level of triglyceride (P=0.001) and cholesterol (P=0.001) as compared to control. The p53 codon 72 polymorphism analysis on hyperlipidemic patients shows higher percentage of Arg homozygous genotype and lesser percentage of Arg-Pro heterozygous genotype. None of the samples exhibited proline homozygous genotype. Conclusion: This study concludes that p53 codon 72 polymorphism can be used as a marker for early diagnosis of hyperlipedimia leading to cardio vascular disease.

Silver Nanoparticles - Potential Antiplatelet / Antithrombotic Agents

Debapriya Bandyopadhyay* and B K Gupta** *Assistant Professor, **Professor and Head, * & ** Department of Biochemistry, Subharti Medical College, Swami Vivekanand Subharti University, Subhartipurum, N.H. 58, Delhi­Haridwar Bypass Road, Meerut­250005 Email: [email protected] Objective: We prospectively evaluated Silver nanoparticles as potential antiplatelet / antithrombotic agents. Methods: Platelets were isolated from anticoagulated human whole blood sample from fifty healthy donors. These platelets were suspended in physiological buffer and each sample was divided into four tubes, in three of them various concentrations of prepared and characterized Silver nanoparticles (SN) were added, fourth tube served as control (C). Following platelet function studies were done in all the four platelet samples: Thrombin induced aggregation (TIA) Integrin mediated platelet adhesion on immobilized matrices (IPA) Integrin mediated cell signaling and associated cytoskeletal changes- spreading and degranulation (ICS) Clot retraction (CR) Results:TIA was increasingly less marked with increasing concentration of SN as compared with

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C. SN showed concentration dependant Thrombin induced Platelet Aggregation inhibition. IPA and ICS were also increasingly less marked with increasing concentration of SN when compared with C. SN decreased platelet adhesion and Integrin mediated cell signaling in a concentration dependant manner. Similar were the observations for CR. Increasing concentration of SN resulted in incremental inhibition of Clot retraction. Conclusion: Silver nanoparticles appear to be a potent anti-platelet agent even in low concentrations. It has the potential for maintaining a lower activation state for platelets, decreasing platelet aggregation and their adhesion to Fibrin clot. With these observations we concluded that Silver nanoparticles are potential antiplatelet / antithrombotic agents. Nano Silver maybe used in therapeutics after appropriate trials and can find a place among antiplatelet agents in use now.

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Abstract No: 8.6

Rapid Detection of Mutation in RRDR of rpo B Gene for Rifampicin Resistance in MDR-Pulmonary Tuberculosis by DNA Sequencing

Dr Surajeet Kumar Patra*,Dr Anju Jain*,Dr B L Sherwal** and Dr Ashwani Khanna *** *Department of Biochemistry,**Department of Microbiology, Lady Hardinge Medical College & Associated Hospital, New Delhi, India, ***Department of Chest Clinic, Lok Nayak Hospital, New Delhi, India Email id ­ [email protected] Objective: To detect the site of mutation in RRDR Results: We observed three different types of of rpo B gene for rifampicin resistance in MDR- mutation in the RRDR of rpo B gene. The TB by DNA sequencing. frequency of mutation in codon 531(TCG!TTG), 526(CAC!TAC) and 516(GAC!GTC) are 60 Methods: 50 MDR-TB patients were enrolled in %, 26.6 % and 6.6 % respectively. Of the total our study after informed written consent. cases studied, 6.6 % cases, although resistant to Mycobacterial DNA was extracted from sputum rifampicin, did not show any mutation in the samples by Universal Sample Processing (USP) RRDR of rpo B gene. method and RRDR of rpo B gene was amplified by PCR using primers RP4T and RP8T and then Conclusion: Codon 531(TCG!TTG) is the most sequenced by automated DNA sequencing. The common site of mutation in RRDR of rpo B gene nucleotide sequences of RRDR of rpo B gene were for rifampicin resistance in MDR-pulmonary compared with the reference sequence. tuberculosis followed by codon 526(CAC!TAC) and codon 516(GAC!GTC). Abstract No: 8.7

Apo E Genotyping from Blood Stored on Filter Paper

Rizwana Quraishi1, Ramakrishnan Lakshmy2, Ashok K Mukhopadhyay3 and Bansi L Jailkhani4 National Drug Dependence Treatment Centre1, Departments of Cardiac Biochemistry2 and Laboratory Medicine3, All India Institute of Medical Sciences, New Delhi, India Hamdard Medical College4, Jamia Hamdard, New Delhi, India Objective: With increasing incidence of lipid derangements among Asians it becomes imperative to identify genetic markers that may predispose individuals to coronary events. Apo E is one of the well recognized genetic factors. In the present study blood spotted onto filter paper is used as a transport system to study genotyping of Apo E gene. fresh as well as filter spotted blood using previously described methods.

Results: The result obtained from filter spots showed exact match of the genotyping when compared to fresh blood. The study was further validated using samples collected on to filter paper from four centers (Delhi, Hyderabad, Bangalore and Lucknow) and stored for eight years at room Methods: Fifty five samples were collected on to temperature. The genotyping was done filter paper with their corresponding blood samples successfully along with the quality checking. coming to AIIMS for lipid investigation. The filter discs were stored at room temperature for storage Conclusion: The present study shows that dried study. The DNA samples were extracted from the blood stored up to eight years, collected from

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temperature conditions varying between 20 to 45 degrees centigrade with humidity upto 16 to 80 percent can be used for genotyping. Since field conditions in developing nations are not well equipped for performance of the test spot, this

study would help in feasible blood samples collection and presenting their stability thus facilitate the exchange of DNA samples among researchers

Abstract No: 8.8

Angiotensin Converting Enzyme Gene Insertion Deletion (I/D) Polymorphism and its Association with Microalbuminuria in Essential Hypertension

K.Ramalingam *, K.Santha , S.Sethupathy and R.Vinoth Kumar * *Department of Biochemistry, Narayana Medical College and Hospital, Nellore ,A.P. Division of Biochemistry, Raja Muthiah Medical college and Hospital, Annamalai University, Annamalai Nagar, TamilNadu. Background: Essential hypertension is a known risk factor of Cardiovascular and Reno vascular dysfunctions. The rennin-angiotensin- aldosterone system ( RAAS) plays a significant role in the development of target organ damage in essential hypertension. Micoalbuminuria has attracted attention as an early marker of organ damage in hypertension. Angiotensin converting enzyme (ACE) gene I/D polymorphism is aasociated with target organ damage in essential hypertension. Objective: The aim of the present study is to know the distribution of ACE (I/D) polymorphism and its relationship with microalbuminuria in essential hypertensive patients. Methods: 208 clinically diagnosed essential hypertensive patients without any associated diseases and 220 age and sex matched healthy control subjects were included in this study. EDTA blood was collected for the extraction of genomic DNA and PCR was performed for analyzing Insertion (I) and Deletion (D) polymorphism at the 287 basepair Alu repeat sequence in the intron16 of ACE gene followed by agarose gel

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electrophoresis and results were recorded in the Gel documentation system. The possible ACE genotypes are D/D, I/I homozygous and I/D heterozygous. Morning random urine samples were collected, microalbumin and creatinine were estimated and results are expressed in micro albumin creatinine ratio. Results: The distribution of D/D genotype of ACE gene was significantly higher (38.9%) in patients when compared to control subjects(20%)Odds ratio 2.5 ( p <0.001) . D/D genotype is strongly associated with microalbuminuria when compared to I/D and I/I genotypes of essential hypertensive patients. Conclusion: The results of this study suggest that deletion (D/D) polymorphism of the ACE gene might be an independent risk factor the development of microalbuminuria which leads to organ damage in essential hypertensive patient. Therefore determination of ACE genotype may be useful for early identification of patient at increased risk for development of target organ damage.

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Abstract No: 8.9

Association of Antioxidant Enzyme Gene Polymorphism in ATT Induced Hepatotoxicity in North Indians

S.V. Rana, Sanjeev K. Sharma, R.P. Ola, S.K. Sinha, J. Kaur, and K. Singh. Department of Super Specialty Gastroenterology, Postgraduate Institute of Medical Education and Research, Chandigarh. 160012, India Background: Drug induced hepatotoxicity in general, is involving toxic metabolite generation and its detoxification in the liver. Glutathione (GSH) plays an important protective role as intracellular free radical toxic reactive metabolites that are generated from biotransformation of drugs and xenobiotics. It's plays an important role in preventing ATT-induced hepatotoxicity. Deficiency in GST activity, because of homozygous null mutations at GSTM1 and GSTT1 genes may susceptibly to ATT-induced hepatotoxicity. polymorphic analysis for GSTM1 and GSTT1 genes were using multiplex PCR method with âglobulin gene as positive control. The presence or absence of GSTT1 (459 base pairs) and GSTM1 (215 base pairs) amplicons was determined in the presence of the control â-globulin gene (268 base pairs).

Results: Mean age of 100 patients was 45.6 years (range 18-70 years). Out of 100 patients, 16 (16%) were ATT-induced hepatotoxicity and 84 (84%) were of non-hepatotoxicity. Out of 16 hepatotoxicity patients, 5 (31.3%) showed Objective: Therefore, the multiplex PCR was GSTM1, 4 out of 16 (25.0%) GSTT1 and 7 out of used to determine the prevalence of deleted/ 16 (43.7%) GSTT1/ GSTM1/ â-globulin. While13 addited genotypes of both GSTM1 and GSTT1 out of 84 (15.5%) showed GSTM1, 28 out of 84 (33.3%) GSTT1, and 43 out 84 (51.2%) GSTT1/ genes in ATT-induced hepatotoxicity patients. M1/ â-globulin in non-hepatotoxicity patients. Methods: In the present study, a total of 100 north Indian adult patients of tuberculosis on ATT were Conclusion: This study showed GSTM1 was enrolled. DNA used for polymorphic analysis was significantly higher in hepatotoxicity patients as isolated from lymphocytes. The genetic compared to non-hepatotoxicity patients. Abstract No: 8.10

Expression of Myogenin and CD105 in Autologous Bone Marrow Derived Mesenchymal Stromal Cells of Duchenne Muscular Dystrophy Patients.

Dr. Sumit Jhajharia*, Dr Prakash C.Mohapatra*, Debashish Das# and Sujay Singh # * S.C.B Medical College, Cuttack, Orissa, # Imgenex India pvt ltd, Bhubaneswar. E-mail id : [email protected] Objective: Application of Bone marrow derived mesenchymal stem cells represent a promising approach for supporting new clinical concepts of cell based therapy in regenerative medicine. The non hematopoietic component of bone marrow includes multipotent mesenchymal stem cells capable of differentiating into fat, bone, muscle, cartilage and endothelium. The present study was undertaken to assess the expression of Myogenin and CD105 in autologous bone marrow derived mesenchymal stromal cells of Duchenne Muscular Dystrophy patients.

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Methods: Five Duchenne Muscular Dystrophy patients between 12 yrs to 24 yrs were enrolled. With informed consent the bone marrow mononuclear cells were harvested, cultured and assessed for the expression of Myogenin and CD105 using Flourescence Activated Cell Sorter (FACS analysis). Results: A significant increase in expression of Myogenin and CD105 was observed in the cultured Abstract No: 8.11

autologous bone marrow derived Mesenchymal stromal cells of Duchenne Muscular Dystrophy patients. Conclusion- The expression of the above mesenchymal stem cell markers in patients with Duchenne Muscular Dystrophy indicate that autologous transplantation of bone marrow derived mesenchymal stromal cells could be of therapeutic potential.

Somatic DNA Damages in Cardiovascular Autonomic Neuropathy

Supriya Simon A1, Jayapal V2 and Dinesh Roy D3, 1 Dept. of Biochemistry, Pushpagiri Institute of Medical Sciences and Research Centre, Thiruvalla, Kerala ­ 689 101; 2Senior Consultant Cardiologist, Govt. General Hospital, Thiruvananthapuram, 3 Genetika, Centre for Advanced Genetic Studies, Thiruvananthapuram ­ 695 024. Introduction: Cardiovascular autonomic diabetic neuropathy (CADN is one of the most clinically significant complications of diabetes mellitus (DM), but one of the least frequently diagnosed. The pathophysiology range from genetic disorders with specific gene defects to metabolic disorders with accumulation of toxins and to autoimmune disorders. An increased level of micronuclei has been shown to be marker of chromosome damage. Methods: Present study was undertaken to quantify the extent of somatic DNA damages by cytokinesis-block micronuclei (CBMN) assay in subjects suffering with CADN and to asses the various socio-economic, demographic characteristics. Forty six subjects suffering from autonomic neuropathy formed the study groups. All the subjects were suffering from diabetes for atleast 8 years and have varying degrees of Abstract No: 8.12 coronary artery diseases. Twenty five healthy age and sex matched control subjects were selected. Two to three ml of venous blood was collected aseptically in heparinized vacuutainers and CBMN assay was performed. Results: The mean CBMN frequency of 15.21 of the study subjects was significantly higher than the CBMN frequency of 10.6 observed in the control subjects (P<0.05). The mean CBMN frequency in patients with CADN with respect to various risk factors such as dyslipidemia, hypertension, smoking, chewing, and drinking showed increased level of somatic damage. Conclusion: This study clearly demonstrated an increased level of somatic damages in CADN patients which was found to be increasing with risk factors associated with life style and duration of diabetes.

Interaction of Iron Deficiency Anaemia and Alpha Thalassaemia in the Population

Tapash Rudra, Sila Chakrabarti and Bani Sengupta Thalassaemia Counselling Unit, Vivekananda Institute of Medical Sciences, 99 Sarat Bose Road Kolkata 700026 Background: Anemia or hemoglobin deficiency

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in human red blood cells is one of the most

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common clinical manifestations in our country, especially in females and children. Hemoglobinopahies and thalassemias together with iron deficiency are the major causes, the majority are due to iron deficiency anaemia, 1015% are due to beta globin gene defects specially beta thalassemia. The alpha globin disorders are less commonly reported because the diagnosis of alpha thalassaemia is usually missed unless in the severe homozygous form or as Hb Barts. But presence of alpha gene has been found in cases of unexplained anaemias.

were co-related with phenotypic changes as well as with serum iron level.

Results: In our present study we have examined a total of 250 samples. The cases were mostly non tribal Bengali cases referred to our unit for investigation of anaemia. 125 cases had iron deficiency anaemia. In 70 selected cases of microcytic anaemias we found two common alpha thalassaemia mutations. We studied 55 selected referral samples to depict the interaction between two common alpha thalassaemia mutations with existing iron deficiency anaemia in the Methods: The alpha cases therefore, referred to populations. our unit for anaemia were then subjected to Multiplex PCR test for the common alpha Conclusions: So from the present study we can mutations ­3.7 and ­4.2 deletions. The results conclude the incidence of á thalassaemia is fairly high together with iron deficiency anaemias in the population. Abstract No: 8.13

Potential of Mycobacterial Excretory Secretory Protein Antigens (SEVA TB ES31, ES-43, EST-6 and ES-20) as Biomarkers to detect Mycobacterium tuberculosis Bacilli

Anindita M1, Thamke D2, Mendiratta DK2 and Harinath BC1 Jamnalal Bajaj Tropical Disease Research Centre1 & Department of Microbiology2, Mahatma Gandhi Institute of Medical Sciences, Sevagram ­ 442102, Wardha (MS). Email: [email protected] , [email protected], [email protected] Objective: To study the utility of mycobacterial ES-31, ES-43, EST-6 or ES-20 antigen as a biomarker for differentiation of Mycobacterium tuberculosis bacilli from nontuberculous mycobacteria (NTM). Results: The presence of antigen in bacilli using FITC labelled antibody was indicated by green fluorescence on the cell surface while its absence by no fluorescence under microscope. In M.tb H 37 Ra and H 37 Rv bacilli, fluorescence was observed on addition of anti ES-31 and ES-43 antigen to mycobacterial bacilli antibody, while no fluorescence was observed in case of EST-6 and ES-20 antigen. However all the antigens were present in detergent soluble antigen of tubercle bacilli. Fluorescence was not observed for ES-31, ES-43, EST-6 and ES-20 antigen in any of the tested NTM as well as in Escherichia coli.

Methods: Detection and location of ES-31, ES43, EST-6 and ES-20 antigens in bacilli and detergent soluble antigen was explored using Fluorescein isothiocyanate conjugated antibodies against these antigens and nontubercular mycobacterial strains were studied for the presence of ES antigen. Detection of these Conclusion: SEVA TB ES-31 and ES-43 may be antigens was done from M.tb H37Ra and H37Rv used as biomarkers to distinguish M.tuberculosis DSS antigen. bacilli from NTM.

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Abstract No: 8.14

Reliability of Pleural Fluid ADA as a Marker for Exudates

Mr.D.S.S.K.Raju, Dr.D.L.Lalitha, Dr.B.Sreehari Babu and Mr.M.Anil Kumar Department of Biochemistry, MIMS, Nellimarla, Vizianagaram, A.P, Email ID: [email protected] respect of ADA level in pleural fluid, a statistically significant increase (p<0.001) was observed in exudates (60.34U/L±35.14) as compared to pleural fluid ADA level in transudates (14.68U/L±2.85). The pleural fluid/serum ADA ratio is also elevated, which is statistically significant (p<0.001) in exudative cases when compared to transudative cases. In the three groups of exudative pleural Methods: In the present study 35 cases comprising effusion (tuberculosis, malignancy and of tuberculosis, malignancy and parapneumonic parapneumonic effusion), it was observed that effusion were included in the study as exudates in there was significant rise of pleural fluid ADA in which 22(62%) were men and 13(48%) were tubercular effusion (97.37U/L±9.25) cases in women. Another 25 cases of pleural effusion due comparison with cases of malignancy(30.8U/ to hypoproteinemia, liver cirrhosis, and congestive L±1.78) and parapneumonic (28.57U/ cardiac failure constituted the transudates, which L±4.39)effusions. Further a positive correlation included 18(72%) men and 7(28%) women. In all coefficient (r=0.98) was established between the the cases serum and pleural fluid protein as well increase of pleural fluid protein and pleural fluid as serum and pleural fluid ADA were estimated ADA in case of tuberculosis. by Biuret method and Gusti & Galanti method Conclusion: In differentiating exudates and respectively. transudates, estimation of pleural fluid ADA and Results: In the group of 35 patients with exudates, ratio of pleural fluid/serum ADA can be considered pleural fluid/serum protein ratio was significantly as additional parameter of significance higher (p<0.001) as compared to transudates. In Background: Pleural effusion, which is a common clinical entity, requires the identification of the nature of effusion, either exudates or transudates for the diagnosis of the diseases. In comparison to transudate, it is the exudative lesion, which needs precise diagnostic measures to find out the cause of pleural effusion. Abstract No: 8.15

Computational Studies of the Interactions of Some Inhibitors with HIV-1 Protease

Rajkumari Rathore, Savita Rathore, Neha Sharma and Anil Bidwai Department of Biochemistry, Index Medical College, Indore, MP; Email: [email protected] Background: Structure-based design has been an integral player in molecular research for over a decade. HIV-1 protease (HIV-1 PR) is an important target for structure-based inhibitor design of anti-HIV drugs. This enzyme cleaves gag and gag-pol proteins to produce structural and functional proteins in the virus. It has been shown that inhibition of this process produces noninfectious virions. HIV-1 PR is a homodimer, with

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each monomer contributing an aspartic acid to the catalytic dyad. Two flexible flap regions cover the ligand upon binding. Inhibitors of HIV-1 PR are widely used in the treatment of HIV/AIDS. However, due to the rapid development of drug resistance of the virus and side effects encountered upon treatment, new inhibitors of HIV-1 PR are constantly needed.

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Objective: This study was undertaken with the objectives to understand structural features that contribute to the interactions of ligands with HIV1 PR and to elucidate the mechanisms of these interactions. Methods: In this study interactions of three potential inhibitors of HIV-1 PR, namely aha024, xk2 and cu216, were analysed using computational docking methods. A computational docking program, AutoDock, was used to carry out the simulations. AutoDock uses a Monte Carlo simulated annealing technique for configurational exploration with rapid energy evaluation using grid-based molecular affinity potentials. Aha024 docked with a minimum docking energy of -19 Abstract No: 8.16

kcal/mol, rmsd = 0.78 ú and an inhibitor constant of 1.1x10-10 M. The respective values for xk2 were -25 kcal/mol, 0.76 ú and 4.7x10-15 M and those for cu216 were -15 kcal/mol, 0.59 ú and 1.5x10-8 M. Results: Results of docking with these peptidomimetic inhibitors show that the ligands bind in an extended manner: the transition state isosteres bind at the catalytic dyad and the residue side chains occupy pockets on alternate sides of the backbone. The amide bonds engage in a series of hydrogen bonds with the enzyme, and also with a conserved water molecule (H2O 301) positioned between the flap regions and the ligand. This key binding mode difference can provide a unique strategy for nonpeptide inhibitor design

Auto Antibody in The Cases Of Neurological Disorder

Dr.Anand Saran*,Mr.S.S.Haque*,Dr.Ashok Kumar** and Dr.Uday Kumar* *Dept. of Biochemistry,IGIMS,Patna, ** HOD,Dept. of Neurology,IGIMS,Patna Background: An autoantibody is an antibody (a type of protein) manufactured by the immune system that is directed against one or more of the individual's own proteins. Antibodies are normally produced in response to a foreign protein or typically a pathogen, which is a infectious organism. Normally, the immune system is able to recognize and ignore the body's own cells. Sometimes, however, the immune system ceases to recognize one or more of the body's normal constituents as "self," leading to production of autoantibodies. These autoantibodies attack the body's own cells, tissues or organs. The causes of autoantibody production are varied and not well understood. It is postulated that :- 1. Due to a genetic predisposition, 2. Environmental trigger,such as a viral illness or a prolonged exposure to certain toxic chemicals, 3. A hormonal component. A diagnosis of disease can not be made solely due to detection of Autoantibody Methods:The diagnosis starts with a complete medical history and a thorough physical examination and by observing signs and symptoms. A number of diagnostic studies is required to identify a specific disease. Viz. 1.Blood tests to detect inflammation, autoantibodies, and organ involvement 2.X-rays and other imaging scans to detect changes in bones, joints, and organs. 3.Biopsies to look for pathologic changes in tissue specimens. Results: Antibodies against muscle or neuronal ion channels, and against myelin glycolipids are clearly associated with peripheral neurological diseases. Some of these antibodies are also being identified in central nervous system (CNS) disorders. In this article we present autoantibody of significance in Neurological disorder and various methods of its estimation.

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Abstract No: 8.17

Evaluation of Serum Adenosine Deaminase Activity and Lipid Profile in Type 2Diabetes Mellitus

M.M.Suchitra 1, S.Aruna 1, K.V.N.Raju 1, K.Deepthi 1, V.Suresh 2, Aparna R.Bitla 1, and P.V.L.N.Srinivasa Rao 1 Department of Biochemistry 1 and Department of Endocrinology 2 Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh, India, Email: [email protected] Background: An altered immune status has been found in Type 2 diabetes mellitus (T2DM). Adenosine deaminase (ADA), has the highest activity in T-lymphocytes, and is a well known marker of CMI. Type 2 diabetes is associated with systemic low-grade inflammation, and ADA which mediates an inflammatory response is considered an inflammatory marker. Adenosine, the substrate for ADA enhances the effect of insulin action on glucose transport and in the presence of elevated ADA activity, this effect of Adenosine is lost. Dyslipidemia commonly observed in diabetes, is a well established traditional risk factor for cardiovascular disease (CVD). The present study was taken up to evaluate serum ADA activity and lipid profile in T2DM. Methods: 25 patients diagnosed as T2DM within the preceding 2-3 months and 25 age and sex matched healthy individuals were taken into the Abstract No: 8.18 study. Serum ADA was estimated by the method of Galanti and Giusti using Perkin Elmer UVSpectrophotometer. Fasting blood sugar, cholesterol, triglycerides, HDL were estimated on Beckman Synchron CX9 autoanalyzer. VLDL and LDL were calculated using Friedewald's formula. The differences in the means of the biochemical parameters were analyzed using unpaired student's t test. Results: Significant increase in serum ADA, cholesterol, triglycerides, LDL, VLDL and a decrease in HDL were observed in T2DM patients when compared to controls. Conclusion: Elevated serum ADA activity may be an important marker of an altered immune status even in early onset T2DM, and also indicates an inflammatory state. Diabetic dyslipidemia increases the risk of CVD.

Immunodiagnostic Evaluation of Recombinant Filarial Antigen rWbL2.

Mahendra B.Gandhe, B.V.Sivaprasad.,Kiran Pote and M.V.R.Reddy Mahatma Gandhi Institute of Medical Sciences,Sevagram ,Wardha-442102; Email: [email protected] Background: The implementation of DEC based mass drug administration (MDA) has created need more than in the past, for the novel filarial tests to monitor the treated population and to assess the transmission of infection. Recombinant filarial antigens with higher specificity are expected to be superior to native antigens and also help to overcome the practical difficulty in getting suffi232

cient quantity of antigen material. Methods: Recombinant fialarial antigen WbL2 has been identified by immunoscreening W.bancrofti cDNA library with microfilariaemic sera. Monospecific polyclonal antibodies were raised against rWbL2 in BALB/c mice. Filarial antigen and antibody assays were developed us-

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ing rWbL2, antibodies raised against it and IgG fraction of human clinical filarial serum (IgG). A total of 90 sera samples belonging to different filarial groups were screened. Results: Filarial antigen assays using anti rWbL2 antibodies alone or in combination with FSIgG were 100% specific,but yielded low sensitivity of only 40% and 60% respectively to detect microfilariaemic cases. In contrast the filarial IgG4 antibody assay using rWbL2 antigen detected all

the 25 microfilaraemic cases screened giving 100% sensitivity with 100% specificity. However this assay also picked up some of the exposed / prepatent infections by detecting 16% of the endemic normals screened. Conclusion: Recombinant WbL2 antigen based filarial IgG4 antibody assay provides a useful tool to detect infected cases for the prompt initiation of anti filarial therapy and for survelliance of elimination programme

Abstract No: 8.19

Immunomodulatory and Respiratory Burst Killing Activity of Pyrazine-2Carbohydrazide Derivatives

P.B.Miniyar1 and S. J. Makhija2, 1.Sinhgad Institute of Pharmacy, Narhe Road, Narhe, Pune-411041(M.S.), 2. S. B.College of Science, Dept. of Biochemistry, Aurangpura, Aurangabad­431001 (M.S.), E-mail: [email protected] Background: Tuberculosis has been a major health problem for developing countries including India. Multi-Drug Resistant Tuberculosis (MDRTB) is a growing hazard to human health worldwide and threat to control of tuberculosis. Current estimates report the prevalence of primary and acquired MDR-TB in India as 3.4% and 25% respectively. In the last two decades, the AIDS epidemic and the deadly synergy between Mycobacterium tuberculosis and HIV have favored the recrudescence of microbial infections throughout the world. The current recommended strategy for microbial infections is facing two problems: Multi Drug Resistance (MDR) and HIV/AIDS pandemic. Additionally, in patients with impaired cellular immunity (HIV-syndrom), mycobacterial and fungal (Aspergillus, Histoplasma, etc.) infections predominate and may coexist. Emergence of multi-resistant strains of bacteria has increased the problem of drug

resistance dramatically. Every country in the world has drug resistance bacteria. Objective: The development & use of new antimicrobial agents is one of the ways to combat the emergence of anti- microbial resistance. Synthesis of heterocyclic compounds to explore the potential biologically active agents still draws continued interest. Hence the different aldehydes substituted 5-methyl pyrazine/ pyrazine-2-carbohydrazide derivatives were prepared and tested for its immuno modulatory and respiratory burst killing activity. Results : The derivatives PH 9, PH 13, PM 2, PM 5, PM 6, PM 14 have been found to be more promising amongst the compounds tested as compared to the ­ve control.

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Abstract No: 8.20

Serum Lactate ­ Not a Good Marker of Disease Severity in Adults With Plasmodium Falciparum Infection

Dr Rina Triparthy* and Dr Bidyut Kumar Das** *Associate Professor Dept of Biochemistry,SCB Medical College, Cuttack, ** Professor of Medicine, VSS Medical College, Burla; Email-id : [email protected] Objective: Studies indicate lactate level as an accurate prognostic indicator and major cause of death in children with severe malaria.This study was undertaken to assess serum lactate levels in various subsets of severe malaria and its prognostic importance in adults. Methods: Adult patients of severe malaria were examined and classified as : Cerebral Malaria(CM), n = 29 Non Cerebral Severe Malaria(NCSM), n = 58 Muti Organ Dysfunction(MOD) ­ classified as involvement of two or more organs, n = 36 Two control groups were Uncomplicated falciparum infection (UM), n = 64 Endemic Normals (EN), age and sex matched, n = 10 Venous blood collected was used for routine and biochemical tests. Serum lactate was estimated by endpoint colorimetric method. Abstract No: 8.21 Results: Mean control values for EN and UM were 24.4 ± 7.03 mg/dl and 37.01 ± 25.5 mg/dl respectively. In comparison value for CM was 43.8 ± 24.3mg/dl, NCSM was 39.5 ± 28.6 mg/dl and MOD was 58.5 ± 92.3. The values for UC and subsets of severe malaria were significantly high(p < 0.01) compared to EN. There was no significant difference of lactate values between different subsets of falciparum infection (p > 0.05) although values were highest in MOD. The mean lactate levels in survivors was 40.7 ± 29.2 mg/dl compared to 87.7 ± 143.1 mg/dl in those who died but the difference was statistically insignificant(p<0.24). Correlation between serum creatinine and lactate was positive (r = 0.11). Conclusion: Serum lactate levels in adult falciparum infection in Orissa has insignificant prognostic value contrary to observations made in other parts of the world.

Altered Renal Profile and Liver Function Test in Case of Complicated Plasmodium Falciparum Malaria

Shivaraj Gowda, Prakash B Desai, Sindhu J Shetty, Vijayetha S Kagwad, and Mahantesh B Ilakal Department of Biochemistry, J. N. Medical College, Belgaum 590010, Karnataka (India); Email: [email protected] Background: Malaria is one of the major public health problems in India. Around 1.5 million confirmed cases are reported annually by the National Vector Borne Disease Control Programme (NVBDCP) India, of which 40­50% is due to Plasmodium falciparum. Serious

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complications like cerebral malaria, acute renal failure (ARF), severe anaemia, jaundice, acidosis, acute respiratory distress syndrome (ARDS) etc are seen usually in association with falciparum malaria. Severe malaria may present with non specific clinical features making it difficult to distinguish from other febrile illness.

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old male showed renal profile high for Serum Creatinine 1.6 mg/dl and Blood Urea 75 mg/dl. The electrolytes showed low Serum Sodium 133 mEq/L and normal Serum Potassium 4.6 mEq/L. The liver profile was abnormal showing high Serum Total Bilirubin 10 mg/dl (Conjugated 8.3 mg/dl, Unconjugated 1.7 mg/dl), Serum SGOT 138 U/L and SGPT 97 U/L. While Serum Total Protein Results: This is a report of a 50 year old female 6 g/dl, Albumin 2.6 g/dl and A: G ratio 0.8 were showing renal profile with elevated Serum low. Creatinine 2.5 mg/dl and Blood Urea 111 mg/dl. The electrolytes values showed high Serum Conclusion: P.malariae and P.falciparum are Potassium 5.4 mEq/L and normal Serum Sodium responsible for clinically important renal disease 144mEq/L.The liver profile were abnormal the former causes chronic progressive syndrome showing high Serum Total Bilirubin17.1 mg/dl and the latter causes acute renal disease. Jaundice ,(Conjugated 14.7 mg/dl, Unconjugated 2.4 mg/ by itself is not considered as severe malaria. When dl) and Serum SGOT 62 U/L whereas SGPT28 serum bilirubin >3mg/dl is found in association U/L, Alkaline Phosphatase 65 U/L were normal. with other vital organ dysfunction, which indicates Serum Total Protein 6 g/dl, Albumin 2.4 g/dl, A:G severe disease. ratio 0.7 were low. Whereas the report of a 20 year Methods: We report two cases of severe malaria who presented with non-specific clinical features. One of the cases is a 50 year old female who was brought to our hospital with high grade fever and severe abdominal pain, whereas the other case was a 20 year old male presented with high grade fever and headache. Abstract No: 8.22

A Study on Acute Phase Proteins and its Relationship with Hemoglobin in Anemic Pulmonary Tuberculosis Patients

Subodh Kumar1, Kiran Saxena1, Rashmi A Kulkarni1, Avanish Mishra1, Ramesh Pradhan1 , Rosy L. Pradhan,1 and A.R.S Sinha2 1. Department Of Biochemistry, Sri Aurobindo Institute Of Medical Sciences, Indore (MP), 2. Department Of Biochemistry, Peoples Medical College, Bhopal (MP) Background: Tuberculosis (TB) remains one of the world's leading infectious causes of death among adults. Every year, approximately 2 million persons in india develop tuberculosis (TB), accounting for one fourth of the world's new tb cases. Acute phase protein (APP) are a class of diverse proteins whose blood plasma concentration increases or decreases during the response to inflammation, in the acute phase. Objective:Therefore, the present study was planned to know whether there is relationship between serum ferritin and haptoglobin with hemoglobin in anemic pulmonary tuberculosis patients when compared with healthy control of the same age group (20-58yrs) and sex. Methods: A prospective study was carried out in 40 newly diagnosed patients with anemic pulmonary tuberculosis at Sri Aurobindo Institute of Medical Sciences & from the dots centre, indore. Normal control subject (n=35) were recruited for the study. Serum haptoglobin and ferritin levels were estimated by ELISA method & hemoglobin by cell counter (sysmex kx-21). Results: The above study revealed that there is significant increase in serum haptoglobin & ferritin levels with decrease in hemoglobin concentrations when compared with healthy controls. The study suggests that `rise in acute phase protein causes disturbance of iron homeostasis which leads to anemia.'

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Abstract No: 8.23

Plasma levels of Interleukin-6 and Interleukin-10 in Rheumatoid Arthritis

Vasanthi Pallinti*, Nalini Ganesan* and Rajasekhar G# *Department of Biochemistry, #Department of Rheumatology, Sri Ramachandra Medical College and Research Institute, Sri Ramachandra University, Porur, Chennai ­ 600 116; E-mail ID: [email protected] Objective: To study the plasma levels of proinflammatory cytokine, interleukin-6 (IL-6) and anti-inflammatory cytokine, interleukin-10 (IL-10) in patients with Rheumatoid Arthritis (RA) and healthy controls. Methods: 120 patients diagnosed with RA (satisfying ARA criteria) were selected for the study. 50 healthy age and sex matched subjects were included in the control group. PotassiumEDTA samples were analyzed for Erythrocyte Sedimentation Rate (ESR, 1 hour) by Westergren method and Lithium heparin plasma samples were analyzed for Rheumatoid factor (RF), interleukin6 (IL-6) and interleukin-10 (IL-10) by ELISA using commercial kits. elevated (62.7 ± 27.9mm) compared to control subjects (19.37 ± 11.4mm). The plasma IL-6 levels were significantly elevated in RA group when compared with control subjects (p<0.001). A significant positive correlation was observed between IL-6 and RF in both RA and controls. A moderate increase in the levels of IL-10 was observed in RA group compared to the controls. Statistical analysis was performed using SPSS (version 15.0) software.

Conclusion: Increased levels of IL-6 and IL-10 in the patient population clearly indicate the role of these cytokines in the pathogenesis of the disease. The increased ESR values and the RF positive status in the patient group indicate active disease condition. The correlation between IL-6 Results: 81% of the RA group was found to be and RF indicates the close association of IL-6 to RF positive in comparison with 4% for the control disease activity in RA patients. group. ESR values of RA group were significantly Abstract No: 8.24

Anti - CCP Auto Antibodies in Rheumatoid Arthritis

Vinod AN*, Nalini Ganesan* and Rajasekar# Department of Biochemistry*, Rheumatologist#, Sri Ramachandra Medical College and Research Institute, Sri Ramachandra Medical University, Porur, Chennai. India Background: Rheumatoid arthritis (RA) is a chronic inflammatory arthritis with potential to cause significant morbidity and mortality. It is important to diagnose RA at a very early stage in the disease. Serological tests are important in diagnosing rheumatic diseases. To facilitate diagnosis during the early stages of the disease a good serological marker is needed. Besides the rheumatoid factor (RF), another group of auto antibodies has recently been detected in serum of

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patients with RA. Anti ­ CCP antibodies are undoubtedly among the most interesting antibodies to have been identified in recent years. Auto antibodies directed against citrullinated proteins have a much higher specificity for RA. These citrullinated proteins have been detected in the synovium of RA patients. Citrulline is a non standard amino acid being formed as a result of post translational modificational change of protein bound arginine by peptidyl arginine deaminase.

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Our previous studies on the serum biochemical marker of RA reports 56% of RF positivity in a group of rheumatoid arthritis patients. Several observations have indicated that anti-CCP positive in early RA patients may develop a more erosive disease than those without anti ­ CCP. In this review we will discuss the process of formation of citrullinated protein, its transformation to auto Abstract No: 8.25

antigens and the clinical utility of these auto antibodies. Objective: We have planned to estimate the anticcp antibodies in the RA patients (early, established and late disease) to study our observations on the clinical utility of this autoantibody in RA.

Molecular Mechanisms Underlying the Effect of Papaya Seed Products in Regulating Male Fertility

N. Pathak, R. M. Samartha, D. Jain, K. K. Maudar, S. Tiwari, S. Gupta. and P. K. Mishra Bhopal Memorial Hospital & Research Centre, Raisen Bypass Road, Bhopal Background: Fertility regulation with plant preparation/medicaments has been mentioned in ancient texts of indigenous system of medicine. The major advantage of the use of herbal drugs, being non-steroidal is that they have least side effects, having low toxicity potential for human use in the history of folklore medicaments and more importantly cost effective. Plant derived compounds or their derivatives are the basis of a large number of established drugs. Approximately 80% of the world's population depends on healthcare system that involved the use of traditional medicine, medical plants, including fertility regulating plants, are integral part of such practices. The seed product of Carica papaya has shown promising results in proving contraceptive effectiveness in animal models. However, concerted efforts at a molecular level to decipher the mode of action of the drug have yet not been established. Objective: To elucidate the molecular mechanisms of the contraceptive effects of Carica papya seed products for developing a plant based oral pill for male contraception. Methods: Studies were conducted on GC-1spg cells, a continuous mouse spermatogonial cell line which shows characteristics of a stage between type B spermatogonia and primary spermatocytes. Both dose dependent and time course response (n=3) using active compounds of Carica papaya. The studied compounds CP-M1 & CP-M2 were isolated from methanol sub-fraction of the benzene column chromatographic fraction of the crude chloroform extract of Carica Papaya Linn. Seeds (Honey Dew Variety). Cellular DNA damage response was studied for qualitative phosphorylation states of ATM, ãH2AX proteins and quantitative state of p53 phosphorylation; measure of apoptotic index through annexin-V/PI assay and mitochondrial depolarization; induction of oxidative stress by CM-H2DCFDA and formation of 8-hydroxy-2'-deoxy guanosine; levels of anti-oxidant defense system enzyme glutathione reductase, prior to and following treatment were also investigated. Results: Both the compounds exhibited an increasing trend in DNA damage responsive parameters in treated cells in comparison with controls. CP-M1 & CP-M2 exposed cells showed an increased mitochondrial depolarization and intracellular ROS generation due to oxidative burst, with altered antioxidant defense mechanism finally leading to an inexorable cellular demise as implicated by the higher apoptotic index in studied cells.

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Conclusion: Results obtained so far from the However, studies aimed at understanding signaling preliminary studies conducted demonstrate that mechanisms involved in DNA damage, apoptosis CP-M1 & CP-M2 compounds act by negatively and cell cycle regulation are in progress. regulating the DNA damage signaling pathway. Abstract No: 8.26

Prevalence of HCV Genotypes and Significance of TH1/TH2 Cytokines in Response to Combination Therapy: A Pilot Study from Central India

P. K. Mishra, A. Bhargava, R. P. Punde, S. Tiwari, P. Vashistha, S. Varshney and K. K. Maudar Bhopal Memorial Hospital & Research Centre, Bhopal, India Background: HCV infection is a worldwide public health problem and is a major cause of hepatic fibrosis, cirrhosis, end-stage liver disease and hepato-cellular carcinoma. Till date 6 different HCV genotypes and more than 70 sub-types based on the nucleic acid sequences have been identified. Due to such high levels of genetic heterogeneity, treatment options for HCV infections are poor. At the moment, the only accepted antiviral therapy with proven effectiveness is a combination therapy of (pegylated) interferon alpha and ribavirin. Moreover, the precise immune response against HCV genotypes and their region specific prevalence in central India is unknown and the impacts of viral load, genotype, and immune mediators on the clinical course are poorly defined. Thus, in order to refine the current knowledge of treatment strategies there is an ongoing need to evaluate the influence of host and viral factors. Aim: The objective of present study was to perform a clinical, epidemiological and immunological characterization of chronic hepatitis C infection in a cohort from central India. Methods: A total of 34 patients suffering from chronic HCV were included in the study. Response to combination therapy (IFN-R) was classified as rapid virological response (RVR), defined as undetectable HCV RNA at week 4 after the initiation of combination therapy; early virological response (EVR), defined as undetectable HCV RNA at week, 5 to 12 of combination therapy;

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and late virological response (LVR), defined as undetectable HCV RNA after week 13 of combination therapy. A SVR was defined as clearance of HCV RNA (<600 IU/ml) at 6 months after the cessation of combination therapy. Detection of HCV RNA was done through COBAS AMPLICOR Analyzer (minimum detection limit 600 IU/ml). Genotypic characterization of HCV was done using Linear Array HCV genotyping test kit from Roche Diagnostics, Mannheim, Germany. Analysis of secreted levels of inflammatory cytokines was performed using BDTM Multiplex Cytometric Bead Array (CBA) Human TH1/TH2 kit from BDTM Biosciences, San Diego, USA. Results: The prevalence of genotype 3 was found in 91.18 % cases while 28.8% were found to be infected with genotype 1. In genotype 3 infected patients RVR, EVR and SVR were observed in 74.19 %, 16.12 % and 9.6 % respectively. While in patients infected with genotype 1 RVR and LVR was observed in 33.33 % and in 66.66 % patients. With an intention to treat analysis 100 % and 66.6 % SVR was observed in genotype 3 and 1 respectively. However, SVR was not observed in 1 patient out of 3 in genotype 1 infected population. A significant increase in the levels of Th1 cytokines (IL-2, TNF-á, IFN-ã) following therapy was observed during RVR in comparison to their base line levels (before initiation of therapy). In addition, a week Th1 response observed in the patients were SVR was not achieved further

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indicating a strong correlation of Th1 cytokines genotype 3 is more prevalent in the central Indian population along with traces of genotype 1. Our with the SVR. study also indicated that immune therapeutic Conclusion: Given the lack of an effective response is weak in genotype 1 in comparison to vaccine, optimal treatment of chronic HCV genotype 3. These immune responses are directly infection is now perceived as a `must'. Prior related to the SVR. Besides this a better recognition of the infecting genotype will assist understanding of these interactions will allow the development of increasingly specific immunein scheduling the optimal treatment duration. The based interventions for prevention and treatment present study revealed, for the first time, that of chronic HCV. Abstract No: 8.27

Diagnosis of Mycobacterial Tuberculosis Complex (MTBC) in Endoscopic Biopsy Samples and Paraffin-Embedded Tissue Specimens Using Real Time PCR: Implications in Patient Care

G. V. Raghuram, A. Bhargava, R. P. Punde, S. Varshney, K. K. Maudar and P. K. Mishra Bhopal Memorial Hospital & Research Centre, Bhopal, India Objective: The diagnosis of Mycobacterium tuberculosis complex (MTBC) in endoscopy biopsy samples and formalin-fixed, paraffinembedded surgically resected tissues of the gastrointestinal tract remains a complex issue because the most widely used conventional diagnostic tools, such as culture and acid fast bacilli staining are unable to rapidly detect Mycobacterium tuberculosis with sufficient sensitivity. Using fluorescence resonance energy transfer (FRET) technology based on hybridization probes, we applied a novel technique consisting of an internally controlled quantitative real-time PCR assay that provided a significant improvement in detection sensitivity and quantification. Methods: The study included three groups (A) control (n=24) with no previous signs of MTBC, (B) patients (n=28) with known TB origin and (C) patients (n=50) with clinical and histopathological signs of TB but was culture and AFB negative. The samples were obtained from Medical & Surgical Gastroenterology Departments of Bhopal Memorial Hospital & Research Centre. We extracted DNA using DNeasy Blood & Tissue kit (QIAGEN, Germany) and performed Real time assay using Roche Light Cycler 2.0 with FRET probes for specific amplification of 159 bp region of mycobacterium genome. Results: All the samples (n=24) of group A were found to be negative, while in group B out of total 28 cases studied 27 were found to be positive by real-time PCR. In group C out of total 50 cases studied 18 were found to be positive showing a positivity of 36 %. The overall positive and negative predictive values of the test for clinical TB (group C) were 100 % and 96.9 % respectively. Conclusion: Results of our investigation exhibit that the real time detection technology using FRET probes, has much higher sensitivity for the detection of Mycobacterium tuberculosis complex DNA in tissue biopsy samples and formalin-fixed paraffin embedded surgically resected tissues of the gastro-intestinal tract. However, to establish the superiority of this novel technique for MTBC diagnosis in various latent infectious states, it will be necessary to accumulate data from a larger number of patients with suspected tuberculosis infection

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Abstract No: 8.28

Mediators of Immune System and Their Possible Role in Pathogenesis of Chronic Hepatitis B and C Viral Infections

R. P. Punde, A. Bhargava, S. Gupta, S. Varshney, P. Vashistha, K. K. Maudar and P. K. Mishra Bhopal Memorial Hospital & Research Centre, Bhopal, India Background: The circulating inflammatory biomarkers may contribute to the pathogenesis of chronic viral hepatitis infections. The present study was carried out in order to evaluate the levels of circulating biomarkers in the patients with chronic viral hepatitis of different aetiology and their possible role in the pathogenesis of the disease. Methods: The study was divided into 3 groups i.e. group 1 comprise of patients with chronic HBV infections (n=60), group 2 included patients with chronic HCV infections (n=60) while group 3 consist of healthy age and gender matched controls (n=60). The mean age of all the subjects in the study was 35 years. Total viral load was quantified from the EDTA plasma and correlation studies were performed with circulating Th1/Th2 cytokines; levels of C reactive protein (CRP); Abstract No: 8.29 glutathione reductase (GR) and superoxide dismutase (SOD) and levels of circulating nucleosomes. Results: A strong positive correlation of Th2 cytokines and GR with viral load in HBV and HCV was observed. The levels of circulating apoptotic nucleosomes also correlated with viral load. However, no significant correlation with CRP and SOD was observed. Conclusion: These results can be helpful in designing interventional strategies like anti-viral therapeutic and/or immunotherapeutic strategies to prevent the progression and long term consequence, as well as for effective prevention of chronic infections and thereby contributing towards developing personalized medical care

Electrophoresis and Immunochemical Quantitation of Serum IgG, IgA, and IgM in Multiple Myeloma and Study of Complications in Multiple Myeloma

Dr. Sajitha Krishnan and Toni Thomas Department of Biochemistry, Amrita Institute of Medical Sciences, Kochi. Background: Multiple myeloma is a chronic, progressive and fatal malignant condition in which the fundamental abnormality is a neoplastic proliferation of plasma cells, which infiltrate the bone marrow and often other body tissues. The disease affects man kind almost universally. It representing approximately 1% of all types of cancer. Serum protein electrophoresis is the most commonly used diagnostic tool for this disease. Objective: The objective of this study was to evaluate electrophoresis and immunochemical

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quantification of serum IgG IgM & IgA in the diagnosis of multiple myeloma and to study the complication of multiple myeloma with the help of parameters like Total protein, albumin, globulin, A G ratio, Creatinine, Serum Alkaline Phosphatase Serum calcium and Serum LDH. Methods: The study was carried out at department of biochemistry AIMS, Cochin during the period January 2008 to January 2009. Electrophoresis was carried out for the blood samples come with order of serum protein electrophoresis.45 M band

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positive (monoclonal protein present) blood samples were selected and performed immunochemical quantification of serum (IgG IgM & IgA) ,Total protein, albumin, globulin, A G ratio, Creatinine, Serum Alkaline Phosphatase Serum calcium and Serum LDH. Result: Of the 45 M band positive patients, 28 (62.22%) patients of them showed an increase in IgG, 2(4.45%) showed an increase in IgM, 4(8.89%)patients showed elevated levels of IgA, and 11 (24.44%) of them showed low levels of Immunoglobulins(IgG, IgM, IgA). In the study group, about 44% showed increased protein levels, 62% patient's serum globulin levels were above normal.About 42.5% patients showed low serum albumin levels. The A/G ratio was reversed in about 75%. 35.5% of the patients showed an increase in serum creatinine. Among the creatinine elevated patients, 75% of them were under IgG type Myeloma group & about 6% were under IgA type Myeloma and 19% of them in the Light chain multiple myeloma group. Of the 45 patients about 33.33% of the patients have high serum ALP level. About 60% high ALP values belonged to IgG myeloma group, about 13.34% belonged to IgA, and the rest 26.66% belonged to light chain multiple myeloma group. High Serum Lactate dehydrogenase (LDH) is seen in 57.14% patients.Of the 57.14% LDH elevated patients, Abstract No: 8.30

75% high LDH values belonged to IgG myeloma group, about 12.5%belonged to IgA, 6.25%belonged to IgM Myeloma and the 31.25% belonged to Light chain multiple myeloma group. Calcium levels were increased in less than 10% patients. Among the hypercalcemic patients 50% were in light chain myeloma. Conclusion: Serum protein electrophoresis is not sufficient to diagnose multiple myeloma because the position of the band only suggests the type of antibody. Serum protein electrophoresis alone will be insufficient especially if the multiple myeloma is of non-secretory type. In such cases urine protein electrophoresis should be done to detect the light chain produced. Immunoturbidimetric quantitative measurement of immunoglobulin levels used along with serum protein elcetrophoresis can be used as a confirmatory test in the diagnosis of myeloma. So a combination of serum protein electrophoresis and immunoturbidometric quantitative estimation of gammaglobulins along with clinical symptoms can be used as tools for diagnosis of different kinds of multiple myeloma and to classify it. In the present study it has been noted that IgG myelomas showed the highest prevalence rate. Moreover, complications in myeloma were also maximum in the IgG myeloma class. This may be due to the greater percentage of IgG producing B-cells in the human body. Thus these B-cells have a greater tendency to be affected by myeloma.

Comparative Evaluation of Recombinant WbSXP-1 Antigen and B.Malayi Microfilarial Antigens in the Detection and Management of Bancroftian Filariasis

Partha Sarathi Pal, Gajalakshmi Dakshinamoorthy, Sivaprasad BV and MVR Reddy Department of Biochemistry& JB Tropical Disease Research Centre, Mahatma Gandhi Institute of Medical Sciences, Sevagram 442 102, Maharashtra; Email for sending acceptance: [email protected] Background: Immunomonitoring followed by treatment with DEC was found to be very effective in the management of Lymphatic filariasis infection. Hence mass chemotherapy programs using DEC to eliminate lymphatic filariasis required specific, sensitive and cost effective methods for the evaluation of infection status. Antibody assays based on crude filarial extracts are limited by cross-reactions with other nematode antigens and the difficulty in getting sufficient

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parasite material. These problems could be overcome by replacing the native filarial antigens with the recombinant filarial antigens.

ES & rWbSXP-1 antigens showed 92, 64 and 100% sensitivity for asymptomatic mf carriers respectively. Further immunomonitoring of clinical filarial cases with these assays showed that Methods: In this study the diagnostic efficacy of the recombinant WbSXP-1 antigen based assay is recombinant WbSXP-1 antigen and two native more sensitive and specific. filarial antigens viz., Bm mf ´S' & Bm mf `ES' were comparatively evaluated for the detection of Conclusion: In conclusion, it can be envisaged actively infected Cases and their that rWbSXP-1 antigen based filarial IgG4 immunomonitoring followed by DEC therapy. antibody test will be a very useful tool in the detection and management of Bancroftian filariasis Results: Performance characteristics analysis of filarial IgG4 antibody against Bm mf ´S', Bm mf Abstract No: 8.31

Oxidalitive Stress and Immunity Status in Leprosy

V.B.Londhe1, Sevesina P Madhale2, Z.G.Badade3, PE Jagtap4, SK Ahaley5 ..and M. M Chavan6 Asso, Prof JNMC Belgaum1 Sr. Research follow in immunolgy2 prof & Head Dept. of Biochemistry MGMC Navi munbai3 Prof. & head Dept. of Biochemistry B.V.MC, Miraj4 , prof of Biochemistry, BVMC Miraj5 medical superintendent TLMH. Belgaum6 Background: In many infactious and disease Conditions people are severly affected by health related immunity. The lymphocyte is primary cell for immunity system. `T' Lymphocytes, thymus derived responsible for cell mediatory and `B' lymphocytes from bone marrow responsible for humoral immunity in human. Leprosy is a disease, which strikes fear in society as a mutilating, contagious, delaying for treatment due to fear of rejection from family and society, horrifying nature of physical disfigurement and no cure being available until twenty century. Objective: The aim of our study was to explore the role of immunity status as upgrading disease. Methods: In present study fifty clinically diagnosed leprosy patients and fifty age, sex Abstract No: 8.32 matched healthy subjects were selected from civil hospital Sangli, Richardson leprosy hospital Miraj and The leprosy mission hospital, Hindalga Belgaum. Serum Zinc estimated by AAS, Salivary IgA estimated by Elisa, ADA activity estimated by K. Satoh method and bacillary index decided by ear lobe smear, Elbow smear and Tissue biopsy. Conclusion: In our study as upgrading disease, due to oxidative stress with increase in bacillary, index, increases LPO and for defence mechanism and ulilization purpose, serum Zinc, ADA and Salivary IgA activity, which `T' cell functioning cell immunity reflect downgrading concentration as per severity of disease. The estimations of serum Zinc, ADA, salivary IgA and LPO as biochemical markers for immunity in leprosy.

Real Time PCR Quantitation of Minimal Residual Disease (MRD) in Acute Lymphoblastic Leukemia (ALL).

KJ Suneetha, K Nirmala Nancy, KR Rajalekshmy, TG Sagar and T Rajkumar Background: ALL is the most common childhood malignancy constituting about 30% of all

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childhood cancers. Though risk adapted chemotherapy resulted in favourable outcome in the majority of the children, a significant percentage of both children and adults eventually relapse. Such failure is presumably due to residual leukemic cells resistant to standard therapy. A more sensitive and precise assessment of residual leukemic cells would help tailor treatment and improve clinical outcome of childhood ALL . Quantitation of MRD by real time PCR allows the detection of one leukemic cell in 104 to 106 normal cells.

identified by PCR-heteroduplex analysis, ASO designed and MRD in followup samples quantitated by Taqman chemistry using real time PCR . Results: MRD was quantitated in 12 patients. 5/ 12 are in remission and 7/12 relapsed. A maximum detection limit of 10-5 . Ig/TCR rearrangements were found to be reliable markers for MRD detection and relapse prediction. We were able to antedate molecular relapse 10-50 days before the clinical and hematological Relapse.

Methods: 2ml of Bone Marrow and 8ml of Conclusion: Quantitation of MRD by Real time Peripheral Blood at diagnosis and 2ml of BM at PCR ia a specific and sensitive method to antedate different treatment intervals were collected. relapse. Leukemia specific Ig or TCR rearrangement was Abstract No.8.33

Role of GST Polymorphisms in the Susceptibility and Outcome in Acute Lmphoblastic Lukemia

KJ Suneetha, K Nirmala Nancy, KR Rajalekshmy, TG Sagar and T Rajkumar Background: ALL is the most common pediatric malignancy constituting about 30% of childhood cancers. The origin of this disease may be explained by a combination of genetic and environmental factors. The glutathione S transferase family is involved in the metabolism of a wide range of chemicals including environmental carcinogens, reactive oxygen species, and chemotherapeutic drugs. Polymorphisms in GST enzymes have been implicated in the susceptibility and outcome in several cancers. by real time allelic discrimination assay for GSTP1 polymorphisms. Results: We found a significant increased risk for ALL with the GSTM1 null genotype (OR: 1.96, 95%CI=1.08-3.57). Though GSTP1 by itself is not associated with the susceptibility to ALL, when it is combined with GSTM1, a significant increase in risk has been observed (OR=2.78: 95%CI=1.166.69). Kaplan Meier survival analysis shows significant inferior outcome in the patients harbouring GSTP1 val allele (log rank p-value of 0.0025).

Methods: A total of 92 immunophenotyped ALL Conclusions: GST polymorphisms play a patients and 150 cord blood samples(controls) significant role in the susceptibility and outcome were analysed by PCR for GSTM1, GSTT1 and in Acute lymphoblastic leukemia.

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Abstract No: 9.1

CSF Calcium and Magnesium (Ionic and Total) Levels: a Preliminary Study

Asharani.N***.Sindhu, Samson Sujit Kumar G*, Ramakrishna** and Ramalingam K*** Departments of Neurosurgery*, Anesthesia** & Biochemistry***, Narayana Medical College, Nellore-2, A.P Background: There have been only few studies to estimate the normal ionic magnesium levels in the CSF in human beings. Role of serum ionic magnesium has been studied in various cardiovascular, obstetrics and neurological disorders. Before estimating CSF ionic magnesium levels in various neurosurgical disorders we wanted to establish normal range of ionic magnesium in the CSF. Methods: Patients undergoing lumbar puncture for anesthesia and suspected neurological disorders were included in the preliminary study (with ethics committee approval and patient consent). CSF (about 0.5ml) was collected from these patients and sent to the lab where the ionic levels of calcium and magnesium were estimated Abstract No: 9.2 along with total levels. Total Ca++ and Mg++ were measured by photometrically.The ionic Ca ++ measure by ion selective method. The ionic Mg++ measured by atomic absorption spectrophotometer. Results: CSF was collected from 15 patients undergoing LP for spinal anesthesia. The average value of CSF total calcium and magnesium was 5.4mg% (2.64meq/L) and 2.7mg % (2.22meq/L). The average ionic concentration of calcium was 1.38 meql/L for calcium and 1.96 meql/L for magnesium. The ionic magnesium level accounts for significant amount of total CSF magnesium. Conclusions: Our preliminary data suggests that ionic magnesium and ionic calcium contribute to significant proportion of total CSF values.

Serotonin, Dopamine and Norepinephrine Functional Regulation of DA D1 Receptors in Unilateral Rotenone lesioned Parkinson's Rat Mdel.

Jes Paul, Nandhu M.S, Korah P. Kuruvilla and C.S.Paulose Molecular Neurobiology and Cell biology Unit, Centre for Neuroscience, Department of Biotechnology, Cochin University of Science and Technology, Cochin ­ 682 022, Kerala, India. Email: [email protected] Background: Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by selective degeneration of dopaminergic neurons in substantia nigra (SN) pars compacta leading to marked reduction of dopamine (DA) levels in the striatum. Method: The present study analysed the effect of neurotransmitters as treatment on rotenone induced hemi-Parkinson's disease in rats and its

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role in the regulation of DA D1 receptor in the corpus striatum (CS) and cerebral cortex (CC) of the experimental rats. Unilateral stereotaxic single dose infusions of 1 µl of rotenone (8µg/µl) were administered to the substantia nigra of adult male Wistar rats. Neurotransmitters ­serotonin (5-HT), dopamine and norepinephrine (NE) treatments were given to rotenone induced hemi-Parkinson's rats.

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Result and conclusion: Scatchard analysis of DA D 1 receptors showed a significant decrease (p<0.001) in the CS and a significant increase (p<0.001) in the CC in the PD rats compared to control. These altered parameters were reversed to near control in the 5-HT and NE treated PD rats and no change was observed in DA treated PD rats. Real-time PCR results confirmed the receptor data. Cognitive and sensorimotor

activities were reduced in PD rats which were reversed by 5-HT and NE. Our results showed 5HT and NE functionally reversed in DA D1 receptors in rotenone induced hemi-Parkinson's rat. This has clinical significance in the therapeutic management of Parkinson's disease. Acknowledgement:This work was supported by research grants from DBT, DST, ICMR, Govt. of India and KSCSTE, Govt. of Kerala

Abstract No: 9.3

Dopamine D1 Receptor Up Regulation In Cerebellum and Brain Stem In Unilateral 6-Hydroxy Dopamine Rat Model: Antagonism By Serotonin And Gamma Amino Butyric Acid

Nandhu. M. S, Jes Paul, Korah P Kuruvilla and C S Paulose* * Molecular Neurobiology and Cell Biology Unit, Centre for Neuroscience, Department of Biotechnology, Cochin University of Science and Technology, Cochin-682 022, Kerala, India. Email:- [email protected] Background: Parkinson's disease (PD) is a severe and progressive neurodegenerative disease characterized by resting tremor, cogwheel rigidity, bradykinesia and loss of postural reflexes. There is degeneration in the central and peripheral nervous systems in Parkinson's disease, predominantly in dopaminergic nigrostriatal neurons. Methods: In the present study, the effects of serotonin, gamma-aminobutyric acid intranigrally to substantia nigra individually on unilateral 6hydroxydopamine infused Parkinson's rat model was analyzed. hydroxydopamine infused Parkinson's rat compared to control. Real Time PCR amplification of Dopamine D1 receptors were up regulated (P<0.001) in cerebellum and brain stem of 6hydroxydopamine infused rats compared to control. Behavioural studies were carried out to confirm the biochemical and molecular studies. Serotonin and gamma-aminobutyric acid supplementation reversed these changes to control. Our results showed that serotonin and gammaaminobutyric acid treatment functionally reversed DA D1 receptor in unilateral 6-hydroxy dopamine infused Parkinson's model. This is suggested to have clinical significance in the management of Parkinson's disease

Results: Scatchard analysis of dopamine D1 receptor binding parameters using [3H]SCH Acknowledgement: This work was supported by 23390 showed a significant increase in Bmax research grants from DBT, DST, ICMR, Govt. of (P<0.001) in cerebellum and brain stem of 6- India and KSCSTE, Govt. of Kerala

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Abstract No: 9.4

Status of Enzymatic and Non-Enzymatic Anti-Oxidant Systems in Patients with Acute Ischemic Stroke

J. Sudha Rani Assistant Professor, Dept of Medical Biochemistry, GVP Biomedical Engineering College for Women, Visakhapatnam, AP. Objective: Oxidative stress is probably involved in neuronal damage induced by ischemiareperfusion. The present study was designed to measure the enzymatic and non enzymatic antioxidant status following acute ischemic stroke patients with large vessel and small vessel infarcts. Methods: The study included 72 subjects in which 30 were controls and 42 were acute ischemic stroke cases. They were diagnosed basing on clinical manifestations and by CT scan of the brain. The cases were further divided into 2 groups. The ischemic stroke patients with large vessel infarcts (Group-L) and small vessel infarcts (Group-S) based on CT scan of the brain. Serum enzymatic antioxidants (superoxide dismutase and glutathione peroxidase) and the non enzymatic antioxidants (reduced glutathione, vitamin E and vitamin C) were estimated in all the subjects by using spectrophotometer. Abstract No: 9.5 Results: Ischemic stroke cases had significantly lower levels of enzymatic and non enzymatic antioxidants, compared to controls which is statistically significant (p<0.05). The Group L ischemic stroke cases had statistically significant (p<0.05) lower levels when compared to Group S ischemic stroke cases. Conclusion: The antioxidant activity of serum may be an important factor providing protection from neurological damage caused by stroke-associated oxidative stress. The present study suggest that reduced antioxidant status may be used as an indirect evidence of oxidative stress induced neuronal damage in acute ischemic stroke which may be useful for monitoring and optimizing antioxidant therapy.

Nitric Oxide and Lipid Profile in Type II Diabetes Mellitus-- A Case Control Study in East Sikkim

Dr A Ghosh, Ms Y Bhutia, Dr T A Singh and Dr M L Sherpa Department of Biochemistry, Sikkim Manipal Institute of Medical Sciences, 5th Mile Tadong, Sikkim737102, Email- [email protected] Objective: Diabetes Mellitus has reached epidemic proportions with an alarming rise in the Asian countries including India. This particular study has been taken up to compare the serum NO level in patients with diabetes and healthy controls and to establish correlation between serum

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nitric oxide level and diabetes mellitus in Sikkim. Methods: Lipid Profile and serum nitrite were assayed in 52 subjects, 26 of which were diagnosed cases of type II Diabetes Mellitus undergoing treatment in SMIMS and 26 were healthy volunteers.

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The statistical analysis was done using SPSS 17 cantly lower in Diabetics in comparison to non and the comparison between cases and control was diabetics (p=0.001). Total Cholesterol was markedly lower in diabetics but this difference was stacalculated by Student's t ­test. tistically insignificant. Results: Most of the patients were above 40 years of age. Mean serum nitrite was lower in diabetics Conclusion: The pattern of findings in our studin comparison to non-diabetics but the difference ies is consistent with the findings elsewhere but was insignificant (p=0.141). Triacylglyerol, LDL not statistically significant. This could be due to were markedly raised in Diabetics but statistically the duration of diabetes and the role of drugs, diinsignificant (p=0.08), (p=0.55). HDL was signifi- etary and lifestyle modification prior to the study. Abstract No: 9.6

Blood Glucose in Small for Gestational Age Infants

Arvindkumar Chowdhary, R Nisarga, C S Muralidhara Krishna and K L Mahadevappa Dept. of Pediatrics and Dept. of Biochemistry, Kempegowda Institute of Medical Sciences, Bangalore. Background: Small for gestational age (SGA) babies have a higher metabolic rate than the appropriate for gestational age (AGA) infants of the same size and are more prone to hypoglycemia that can result in brain damage,mental retardation and cerebral palsy. Methods: Present study is an assessment of the frequency of hypoglycemia and its clinical manifestations in SGA neonates. Singleton live born babies, born at KIMS Hospital, who were below the tenth percentile were taken as SGA infants. Randomely chosen 200 cases each of SGA & AGA infants in which 52% were male, 54.5% were of gestational age of < 37 weeks and 50% were in the birth weight ranging from 15002400gs. Blood sugar was estimated on a routine chemistry analyser in cord blood just after birth and cubital venous cord blood at 2--3hrs, 6hrs, 12hrs and 24hrs of life. Resulst: Mean blood sugar level at 0 hr and 2-- 3hrs of age in different groups is as follows: 0 hr2--3hrs, SGA preterm infants-- 51.90mg% --44.75 mg% SGA fullterm infants-- 54.13mg% -- 47.07 mg% SGA male infants -- 55.00mg% -- 47.00 mg% SGA female infants--50.00mg% -- 44.oo mg% --

SGA infants weighing less than 1500gs 52.77mg% --46.15mg%

SGA infants weighing more than 1500gs -- 53.07mg% --46.01mg% Mean blood glucose level at 0 hr of age is nearly normal in both SGA and AGA babies but the glucose level falls to hypoglycemic levels within 2-3hrs of extrauterine life more commonly in SGA babies (46.02 mg% in SGA babies as against 51.62 mg% in AGA babies).Hypoglycemic babies presented themselves with signs and symptoms such as lethargy, convulsions, apnea, refusal to feed, tachycardia, jitteriness and the state of collapse. Conclusion: Thus the study suggests that screening of all babies for hypoglycemia at 0 hr, 2-3hrs, 6hrs, 12hrs and after, if necessary, of birth should be mandatory and babies at the risk of developing hypoglycemia should be identified. Early intervention with 10% dextrose helps to alleviate the neurological sequale and prevents the burden of mentally and /or physically impaired children.

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Abstract No: 9.7

Insulin Resistance and Lipid Profile in Pregnancy Induced Hypertension

Asmathulla S. and Rajagovindan D Jakanattane Department of Biochemistry, Sri Manakula Vinayagar Medical College & Hospital. Results: Fasting insulin and HOMA-IR are significantly increased in PIH cases compared to controls. Among PIH cases, Preeclampsia showed significant increase in Insulin and HOMA-IR levels compared to controls and gestational hypertension. Total cholesterol, LDL cholesterol Methods: Twenty eight PIH cases and ten normal and triacylglycerol are elevated significantly in healthy pregnant controls are recruited for this PIH. The preeclamptic cases are associated with study. Serum Insulin levels are measured by significant rise in total cholesterol, LDL ELISA method. HOMA-IR is calculated by cholesterol, VLDL cholesterol, and triacylglycerol (fasting insulin x fasting glucose)/22.5. Total as compared to controls but these changes are not cholesterol, triglycerides, HDL-cholesterol and significant in gestational hypertension. glucose are measured using enzymatic kits using autoanalyzer. VLDL-cholesterol is calculated by Conclusion: The data from the present study documents Hyperinsulinemia, Insulin Resistance Friedwald's formulae. and Dyslipidemia in PIH patients. Objective: To study fasting insulin levels and lipid profile in Pregnancy Induced Hypertension (PIH) cases and to compare them with normal pregnant controls. To calculate insulin resistance in PIH and to compare them with healthy pregnant controls. Abstract No: 9.8

Thyroid Status in Hospital Based Cases from Rajendra Institute of Medical Sciences, Ranchi

*Dr K. K. Sinha and **Dr. B. R. Ekka *Prof. and Head of Department, **Assistant Prof, Dept of Biochemistry , RIMS, Ranchi, Jharkhand E-mail: [email protected] Objective: The incidence of thyroid disease in the patients referred to the Biochemistry Lab, Department of Biochemistry , RIMS Ranchi was studied by analysis of thyroid hormone by ELISA. Data for prevalence of thyroid disease in patients at Jharkhand has not been analysed and the study was undertaken to generate such data. Past three years data has been analysed to know the age, sexwise and tribal prevalence of thyroid disease, to detect subclinical thyroid dysfunction and thyroid hormone levels. assay (ELISA). The criteria for evaluating the functional status of the thyroid was based on normal values given by ELISA kits ( T3 -0.52-1.85 ng/ml ; T4 male 4.4 - 10.8 µg/dl, female 4.8-11..6 µg/dl ; TSH 0.39 ­ 6.16 µIU /ml).

Results: Three years data (2006, 2007, 2008) has been analysed. Out of a total of 2016 cases there were 1153 with normal thyroid status, and rest 863 with overt or subclinical disease and follow up cases. The preliminary observations are ­ (1) The prevalence of thyroid disease amongst women in Methods: Thyroid hormone tests which included the age group 20 ­ 50 yrs is very high (84.7%). Total T3, Total T4 and TSH were performed by (2) Incidence of overt hyperthyroidism (20%) and the method of Enzyme Linked Immunosorbant hypothyroidism (23%) are almost equal. (3)

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Subclinical thyroid disease is 21% of the total, out of which subclinical hypothyroidism being 84% and subclinical hyperthyroidism being 16%. (4) 157 cases were of tribal population which was 8% of the total, out of which 31% were suffering from Abstract No: 9.9

thyroid disease. (5) The ELISA kits were sensitive enough to detect changes in thyroid hormone levels and concurrent with clinical symptoms. Follow up studies of 91cases suggest the usefulness of these tests in diagnosis and treatment of patients.

Effect of Iron Deficiency Anemia on Glycosylated Hemoglobin Levels in Type 2 Diabetics.

Dr.B.Shanthi M.D Associate professor, Dept. of Biochemistry, Sree Balaji Medical College & Hospital, ChrompetChennai; E mail id- [email protected] Objective: Glycosylated hemoglobin is a very important parameter to measure glycemic control. Diabetes is now a disorder of lower socio economic population too. Hence important variable "iron deficiency anemia" & its effect on glycosylated hemoglobin is studied. 25 Controls - non diabetics aged 25- 50 were estimated for glycosylated hemoglobin & ferritin levels. Glycosylated Hb- ion exchange resin chromatography; Serum ferritin- ELISA.

Conclusion: From this study we could infer that iron deficiency anemia increases glycosylated Methods- Subjects : 100 type 2 diabetic patients hemoglobin levels & this has to be definitely taken aged (25-50) with good glycemic control - into account while interpreting glycosylated Hb. Estimated for glycosylated hemoglobin & ferritin. Abstract No: 9.10

Minerals Status in Thyroid Disorders (Hypo & Hyper thyroidism)

B.Suneel, D.R.Nagendra, K.Ramalingam and J.N.Naidu Department of Biochemistry, Narayana Medical College, Nellore. controls Ca+ levels are (12.96 ± 11.1) Increased P levels are (2.01 ± 1.03) Decreased Mg+ levels are (3.8 ± 1.91) Increased Zn+ levels are (167.42 ± 120.7)Decreased. In Hyperthyroidism compared to controls Ca+ levels are Decreased (6.60 ± 2.78) 2+ ± Methods: The studies includes 25 controls with , P levels are Increased (7.07 + 3.05), Mg levels normal TSH, T3& T4 . 25 Thyroid disorders having are Decreased (1.8 ± 0.87), Zn levels are Increased below 4 years in age group of 30 ­ 48 years both (60 ± 30.86). sex were included TSH, T3, T4 was estimated by Conclusion: In this study the hypothyroid patients using ELISA Ca was estimated OCPC method should increased Ca and Mg levels decreased, P ,Mg2+ GEDTA method, Zn+ Nitro PAPS , P and Zn levels in hyperthyroid patients should Molybdate method. Decreased Ca and Mg levels, increased P and Zn Results: In hypo thyroid patient compared with levels Objective: The Study was under taken to evaluate the changes of minerals in Thyroid disorders (hypo & hyper). TSH, FT3 & FT4 as markers of hypo & hyper Thyroidism & mineral Status Ca+, P+, Mg+, Zn+ are parameters studied.

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Abstract No: 9.11

The Role of Salacia Oblonga in Maintaining Glucose and Lipid Homeostasis in Diabetic Rats

Bhagyajyothi.D °, Dr.Vivian D'Souza ¹, Dr.Poornima Manjrekar ², & Dr.C.V.Raghuveer ³, ° - Senior Research Fellow, Department of Biochemistry, Kasturba Medical College, Mangalore, ¹ Professor& HOD, Department of Biochemistry, Kasturba Medical College, Mangalore, ² - Associate Professor, Department of Biochemistry, Kasturba Medical College, Mangalore, ³ - Professor of Pathology, Melaka Manipal Medical College, Manipal; E mail: [email protected] Objective: To study the effect of a hydroalcoholic root extract of Salacia oblonga on blood glucose, serum Total Cholesterol (TC), Triglycerides(TG), and HDL- Cholesterol(HDL-C) levels on long standing diabetes. Results: It was observed that there was a significant reduction in RBS levels in Salacia treated rats. The rats fed with 50mg/kg/day showed 28% decrease in RBS (P= 0.018), whereas the group treated with 100mg/kg/day showed 18% reduction (P= 0.012). Either of the groups did not Methods: Streptozotocin (STZ) induced diabetic show significant change in the lipid profile. The Wistar rats of either sex were treated with two oral plasma TC, TG and also HDL-C levels were doses of Salacia oblonga extract, 50mg/kg/day and slightly low in 50mg/kg/day group, but did not 100mg/kg/day for a period of 16weeks. Random attain statistical significance when compared with blood sugar (RBS) was estimated on Day-1 and at diabetic controls. the end of 16 weeks (previous day of estimating the lipid profile) by ACCU-CHEK Active Blood Conclusion: The hydroalcoholic root extract of Glucose Monitor from Roche with disposable Salacia oblonga lowers the RBS in diabetic rats. strips from the same company. The fasting serum Its effect on lipid profile is not significant on long TC, TG and HDL-C were estimated at the end of standing diabetic rats. There was no appreciable 16 weeks in the Autoanalyser, HITACHI-917 using effect of higher dosage (100mg/kg/day) over 50mg/kg/day. Roche Kits. Abstract No: 9.12

Glycemic Control and Complications of Diabetes Mellitus

Adiga Usha 1 and Adiga MNS2 1 Assistant Professor, Biochemistry, KMC, Manipal, Manipal Objective: According to the American Diabetes Association's Clinical Practice recommendations, monitoring of glycemic status is considered a cornerstone of management of diabetes mellitus in order to minimize the complications. The objective of our study is to find the relationship between the glycemic status and common complications seen in them.

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2

Associate Professor, Pharmacology, KMC,

Methods: One hundred and five diabetic patients receiving treatment were seen over a period of one year. Among these, patients with diabetic nephropathy, retinopathy, diabetic ulcers and urinary tract infections were segregated. Fasting, postprandial blood sugar (monthly) and glycosylated hemoglobin (once in three months) were estimated. The analysis was carried out by means of Graphpad statistical package.

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Results: A total of 60 patients had the above complication (15 each). The average duration of the diabetes was 8.65± 1.26, 10.37± 1.25, 13.23±1.75 and 8.86±0.87 years respectively in patients with diabetic retinopathy, nephropathy, diabetic foot ulcers and urinary tract infections. We found that the highest FBS, PPBS and glycosylated hemoglobin was seen in patients who had diabetic retinopathy (160.67±10.32, 221.50±9.0, 8.04±0.87 respectively). The lowest values were seen with urinary tract infection Abstract No: 9.13

(144.17±11.0, 213.50±5.68, 7.77 ±0.47). There was a statistically significant difference between these groups (p<0.05). The re was no much difference between the other two parameters. Conclusion: - We conclude from our study that urinary tract infections are commonly seen with earliest alternation of blood sugar. Retinopathy was seen in patient with high blood sugars and glycosylated hemoglobin values though it has manifested early compared to other complications.

Relationship of Staple Diet and Abnormal Thyroid Function Observed in Outpatients at Ujjain (MP)

Mane A. Y.*, Bhagwat V. R.** Potey G G* and Gaikwad S B** *Dept of Biochemistry, RD Gardi Medical college, Ujjain (MP), **Dept of Biochemistry, SBH Govt Medical College, Dhule (Maharashtra); Email: [email protected] Background: Thyroid function abnormalities are Results: It was observed that mean T3 T4, TSH common in general population. Among several and T /T ratio were higher in mixed diet group 3 4 factors nutritional factors have detrimental effects while T4, TSH/T3 ratio was higher in vegetarian on thyroid functions. group. Mean T3 and TSH/T3 were higher while Methods: This study attempts to probe in to TSH levels were lower in Jowar group. Mean T 4 dietary pattern and thyroid functions in patients and TSH/T4 ratio were lowest in Bajra group reporting in urban hospital at Ujjain city (MP). This is a hospital based study involving 165 whereas it was higher in Jowar group. outpatients with clinically suspected thyroid Conclusion: The results indicate that Jowar as dysfunctions. Thyroid function tests included staple diet appears to promote hypothyroid state circulating levels of free T3, T4 and TSH as determined by micro particulate enzyme while wheat and rice promote hyperthyroid state. immunoassay technique. Abstract No: 9.14

Evaluation of Cardiovascular Risk Markers in First Degree Relatives of Type 2 Diabetes Mellitus

H. Nandeesha, V. Sathiyapriya, Zachariah Bobby, Aparna Agrawal1 and N. Selvaraj Department of Biochemistry and Medicine1, JIPMER, Puducherry. Objective: The objective of the study is to Methods: 29 first degree relatives of type 2 evaluate cardiovascular risk markers in first degree diabetes subjects and 32 controls were enrolled in relatives of type 2 diabetes mellitus. the study. Subjects with history of diabetes,

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hypertension, myocardial infarction, infections and those who are on any kind of medications were excluded from the study. Fasting glucose, lipid profile, insulin, glycated hemoglobin, C-reactive protein and protein bound sialic acid were estimated in both the groups. Results: Insulin, Total cholesterol, triglyceride, glycated hemoglobin, C-reactive protein and Abstract No: 9.15

protein bound sialic acid were significantly increased in first degree relatives of type 2 diabetes subjects compared to controls. Conclusion: Insulin resistance, dyslipidemia and inflammation can enhance the risk of cardiovascular disease in first degree relatives of type 2 diabetes mellitus.

Lipid Profile and Glycemic Status in Non- smoking Type 2 Diabetes Mellitus on Treatment with Oral Hypoglycemic Drugs

J. Sandhya, B.R.Shyam Prasad and U.Satyanarayana Department of Biochemistry, Siddartha Medical College, Dr. NTR University Of Health Sciences, Vijayawada, Andhra Pradesh. ­ 520008; Email: [email protected] Background: Macro and micro vascular complications are the leading causes of mortality and morbidity in type II diabetic patients. These vascular complications are mainly due to dyslipidemia. Objective: The objective of this study was to elucidate glycemic status and lipid profile in non ­ smoking type II diabetics belonging to low socio - economic group and attending Government General Hospital, Vijayawada, and on treatment with oral hypoglycemic drugs. Methods: Seventy patients (aged 35 ­ 70 years) with history of 5 ­ 15 years of type II diabetes and on treatment with oral drugs (metformin, glibenclamide, etc..) and 35 age matched healthy controls were the subjects of this study. Blood samples were collected after keeping the individuals on overnight fast. Serum samples were Abstract No: 9.16 estimated for triacylglycerol, total cholesterol and HDL cholesterol by enzymatic methods using kits. VLDL and LDL cholesterol were calculated by using Friedewald formula. Results: The diabetic patients had relatively higher fasting and post - prandial blood glucose levels compared to controls. Serum triacylglycerol, total cholesterol, VLDL and LDL cholesterol concentrations were also higher in diabetics compared to healthy individuals. Further, serum HDL concentration was lower in diabetics when compared to controls. Conclusion: It appears from this pilot study that type II diabetics of low socio - economic group, mainly depending on the government general hospital for their health care, are not fully under control with regard to glycemic status and lipid profile despite the use of oral drugs.

Thyroid, Lipid and Glycemic Profile in Dyslipidemia and Altered Thyroid Status

Dr.Sridevi , Dr.Vivian D'souza, Dr.Poornima Manjrekar and Dr.Vinit Anand Email ID: [email protected] Objective: To study lipid and glycemic

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abnormalities in subclinical hypothyroidism cases

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hypothyroid group. Both hypothyroid and hyperthyroid pools had higher FPG values, but fructosamine levels were highest in hypothyroid followed by subclinical hypothyroid & hyperthyroid pool. Among dyslipidemic patients, Methods: A total number of 150 subjects, 30 FPG was higher with comparable fructosamine newly diagnosed hypothyroid, 30 newly and lower TSH with normal T ,T . 3 4 diagnosed subclinical hypothyroid, 30 newly diagnosed hyperthyroid, 30 dyslipidemic patients Conclusion: Lipid profile is deranged in both and 30 normal controls between the age 25-55 subclinical and overt hypothyroidism. All classes years of both the sexes were chosen for the study of thyroid abnormalities except subclinical and FPG, fructosamine, fasting lipid profile, hypothyroid showed propensity to develop T3,T4,TSH were done in all subjects. diabetes, fructosamine levels with highest in hypothyroid cases and lowest in hyperthyroid. This Results: The freshly diagnosed hyperthyroid could be because of altered protein metabolism in patients had comparable lipid profile with the hyperthyroid cases. Dyslipidemic pool showed controls except for high LDL values. In highest fasting plasma glucose levels pointing to hypothyroid pool, TC, TG, LDL, VLDL were the possibility of correlation between higher lipid found to be higher & HDL lower than the normal levels and hyperglycemia. Low TSH and pool. Subclinical hypothyroid pool also showed a fructosamine observed in dyslipidemic patients can similar picture except for comparable HDL levels. be attributed to the subclinical hyperthyroid state This pool had lipid status similar to overt altering the aminoacid metabolism. Abstract No: 9.17

& thyroid and glycemic abnormalities in dyslipidemia patients & to compare the same with euthyroid, overt hypothyroid and hyperthyroid subjects.

Incontrovertible Behavior of Oxidative Stress in Etiopathogenesis of Diabetes Mellitus or its Complications is Disputable

P.P. Singh, Anu Chandra, Chetna Vashishtha, Farzana Mahdi and Abbas Ali mahdi1 Department of Biochemistry, Era's Lucknow Medical College & Hospital, Lucknow, (UP), India, 1 Department of Biochemistry, C.S.M. Medical University, Lucknow, (UP), India Background: Unfortunately India topped the list of diabetic population. In the recent years, raised oxidative stress (OS) has been claimed as one of the significant risk factors. The two main proposed hypotheses are: a) raised OS can trigger DM by inducing insulin resistance which is a cardinal feature of DM and b) hyperglycemia raises OS which exacerbate multiple risk cascade resulting in diabetic complications. Our observations do not unequivocally support either of these claims. dant defense system is an essential component of DM or associated complications. Methods: The study included 209 diabetic patients (Type 1-70 and Type 2-139) and 131 matched controls. Serum/RBC lipid peroxidation levels (LPO expressed as nmol/ml), and antioxidant complement was examined.

Results: LPO levels were raised in diabetics (4.95±2.19) as compared to controls (2.88±1.29), DMT-1 patients had slightly but significantly Objective: To evaluate whether raised OS due to higher (5.22±2.24) than DMT-2 (4.67±2.09). Its heightened lipid peroxidation or weaker antioxi253

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level in different DMT-2 groups was: without complications - 3.81±1.32, with cardiovascular disease 4.49±1.78, retinopathy 2.86±2.11, neuropathy 3.84±1.52 and nephropathy 4.70±0.95.Salient findings are a) in none of the groups all patients had raised OS b) DM with complications did not show raised LPO levels than those without complications c) mean GSH levels were slightly but significantly lower in patients but neither it could Abstract No: 9.18

be categorized as depleted or low d) Antioxidant levels in diabetic and control did not offer any conclusive explanation. Conclusion: Though there is compelling evidence that OS can potentiate and contribute to risk in DM or its complications, our data suggest that under clinical setting its role is generally facultative rather than obligatory

A Comparative Analysis of Anthropometric Measurements, Lipid Profile, Fasting Insulin and HOMA2-IR to Identify Insulin Resistance in Overweight Individuals

Dr.M.D.Sreenivasa murthy 1, Dr. Manjunath M Tembad 2, and Dr. Jayaprakash Murthy DS 3 1 Postgraduate student, 2 Professor, 3, Professor & HOD, Department of Biochemistry, J.J.M.Medical College, Davanagere ­ 577004, Karnataka state. Email ID [email protected] Background: Insulin resistance is more common in overweight individuals and is associated with metabolic syndrome which increases one's risk for the development of type-2 diabetes mellitus and cardiovascular disease. using Microsoft Excel® based HOMA-2 calculator.

Results: The optimal cut-off values for identification of insulin resistance in overweight individuals were: 144 mg/dL for triglycerides Objective: This study was carried out to evaluate (sensitivity-66%; specificity-83%), 3.4 for TG/ the ability of anthropometric measurements, lipid HDLc ratio (sensitivity-68%; specificity-90%), profile and fasting insulin to predict insulin and 17µU/mL for fasting insulin (sensitivity-74%; resistance and their correlation with HOMA2-IR specificity-100%). Linear regression analysis in overweight individuals. showed a significant linear relationship between HOMA2-IR and the following parameters: waist Methods: The present study included 120 subjects circumference, BMI, serum triglycerides, TG/ comprising of 60 healthy controls (BMI: 18.5HDLc ratio and fasting insulin (p<0.001). 24.9) and 60 overweight subjects (BMI: 25.0-29.9) in whom the following parameters were measured: Conclusion: Triglyceride levels, TG/HDLc ratio anthropometric measurements (waist and insulin concentration are practical methods for circumference, waist-hip ratio, BMI Prime); identifying overweight persons who are insulin fasting glucose, triglycerides, total cholesterol, resistant. In the absence of a standardized insulin HDLc and LDLc (enzymatic methods by assay, we suggest that the most practical approach autoanaylzer) and fasting insulin (ELISA by Bio- to identify overweight individuals who are insulin rad microplate reader). HOMA2-IR (updated resistant is to use the cut-off values for either homeostasis model assessment) was calculated triglyceride concentration or the TG/HDLc ratio.

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Abstract No: 9.19

Thyroid Hormone Levels in New Borns of Mothers with Gestational Diabetes

1Dr Geetha A, 2Dr Joseph P Thyparambil, 2Dr Sreekumari S, and 3Dr Sulekha B 1Assistant Professor of Biochemistry, Medical College, Kottayam; 2 Professor of Biochemistry S Gokulam Medical College Thiruvananthapuram; 3 Professor of Biochemistry SUT Medical College Thiruvananthapuram. Email address: [email protected] Objectives: The present study was done to assess the glycated haemoglobin levels. The glycated the thyroid status of new borns of mothers with haemoglobin levels were estimated during the third trimester of pregnancy. gestational diabetes. Methods: The thyroid hormones, Tetraiodothyronine, Tri iodothyronine, and Thyroid Stimulating Hormone were estimated in cord blood. The thyroid function tests were done by the ELISA method, the blood glucose by glucose oxidase-peroxidase method and glycated haemoglobin by the cation exchange resin method. Results: In the present study the tri iodothyronine levels in cord blood were found to be much higher than normal range. Normally the triiodothyronine levels are lower in cord blood (4-141 ng/dl) as compared to the adult (70-207 ng/dl). Triiodothyronine levels in cord blood also showed a statistically significant positive correlation with the severity of maternal diabetes as judged by post prandial blood sugar levels and Abstract No: 9.20 Discussion: The increase in Triiodothyronine levels in cord blood may be due to foetal hyperglycaemia and the resulting foetal hyperinsulinsm. In the mother when the blood glucose levels are high the intracellular entry of glucose is low due to insulin resistance. The foetus of a mother with gestational diabetes is exposed to hyperglycemia due to transplacental transfer of maternal glucose .This stimulates the foetal pancreas to produce more insulin. In the foetus,there is no insulin resistance and the increase in both glucose and insulin levels facilitates the entry of more glucose into the foetal cells which may enhance the activity of 5' deiiodinase causing increased formation of T3 from T4.This may be the reason for increased T3 levels in cord in correlation with maternal hyperglycemia

Lp(a) in Hypothyroidism

F.S.Geethanjali* , M.S. Seashadri** and A.S. Kanagasabapathy*** Departments of Clinical Biochemistry*, Endocrinology ** and Former Professor in Clinical Biochemistry*** , Christian Medical College, Vellore. Objective: Thyroid disorders are known to influence the lipoprotein metabolism. In hypothyroidism the plasma concentrations of the LDL fraction is elevated. Since Lp(a) and LDL share the apoB100 in common, this study is to evaluate the Lp(a) levels in Hypothyroid patients and its risk for artherogenesis in them. Methods: Eighty four primary hypothyroid patients attending the Endocrinology Clinic and sixty four euthyroid subjects were included in the study. Serum Lipid profile, thyroid function tests and plasma Lp(a) levels were performed. Results: The thyroid profile showed typical euthyroid and hypothyroid patterns. The mean age

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for both the groups was comparable. Total cholesterol, triglycerides and LDL cholesterol were significantly elevated in the hypothyroid patients. HDL was marginally increased but not statistically significant. The plasma Lp(a) levels were markedly increased in hypothyroid compared to the euthyroid. Conclusion: The hypothyroid induced changes in lipoprotein metabolism could directly or indirectly promote atherogenesis. The elevated cholesterol and LDLc fraction may be due to the reduced Abstract No: 9.21

activity of the LDL receptor. There are suggestions that the Lp(a) concentrations depend on the amount of apo(a) available for binding to apoB100 and not vise versa. The thyroid hormones which are known to affect the biosynthesis at the transcriptional and / or translational level of the apoprotein could reasonably exert an effect on apo(a) synthesis. But it is unclear whether the increased synthesis or the decreased catabolism could be the cause of Lp(a) elevation in hypothyroid patients.

Role of Myelin Oligodendrocyte Glycoprotein and Galactocerebroside Antibodies on Phosphorylation and Degradation of Myelin Basic Protein In Isolated Myelin: Implications in Multiple Sclerosis.

Krishnakumar Menon*# Sidhy Viha C.V*+., Krishna Prasad G.N*+. and Claude C.A. Bernard# *Institute of Molecular Medicine, +Department of Biochemistry, Amrita Institute of Medical Sciences, Elamakkara, Kochi, India and #Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Victoria, Australia. email:[email protected] Background: In Multiple Sclerosis (MS), both neuronal and myelin damage is the cause of the ensuing neurodegeneration leading to neurological deficit. This process results from an autoimmune attack on myelin in combination with genetic susceptibility and the involvement of an unknown environmental factor. Although the pathogenic role of immunoglobulins (Igs) directed to myelin antigens in inducing myelin/ oligodendrocyte damage have been elucidated, how the pathogenic Igs induce biochemical breakdown of myelin is poorly understood. Role of MS Igs as well as monoclonal antibodies (mAbs) directed to myelin oligodendrocyte glycoprotein (MOG) and Galactocerebroside (GalC), in inducing significant myelin basic protein (MBP) degradation mediated by neutral proteases in myelin had been shown. MBP plays a pivotal role in the structural organization of myelin. Thus breakdown of MBP could lead to myelinolysis.

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Methods: In order to elucidate the mechanism by which specific anti-MOG and anti-GalC antibodies induce MBP degradation, we have investigated the possible ways by which MBP is made vulnerable to proteolysis as well as the relationship between Gal C and MOG. Results: In this report, we demonstrate that antibodies to GalC and MOG induced phosphorylation of MBP prior to its degradation in isolated human myelin. Neither such a phosphorylation nor a degradation of MBP could be observed using antibodies directed against MBP or a plant glycoprotein (control Igs). Conclusion: We here suggest that phosphorylation of MBP may result in the exposure of MBP to proteolysis as a result of poor MBP-lipid interaction leading to myelinolysis. It is likely that these demyelinating antibodies may mimic the naturally occurring ligands to MOG/GalC and we propose a possible interaction between MOG and GalC in inducing myelin damage thus exploiting

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Abstract No: 10.1

Glycosylated Hemoglobin Levels in Relation to Antioxidant Status and Lipid Profile in Type 2 Diabetes Mellitus Patients

Prasiddha Tilak1, Mahesh S1, Naureen Anwar1 and Dr Mungli Prakash1 1 Department of Biochemistry, Kasturba Medical College, Manipal, Karnataka-576104, E-mail for correspondence: [email protected] Objective: Type 2 Diabetes Mellitus (Type 2 DM) is a metabolic disorder related to either insulin deficiency or resistance. Glycosylated Hemoglobin (HbA1c) is used to monitor long term regulation of blood glucose. Studies relating HbA1c with antioxidant status and lipid profile are very few. Present study was carried out to determine the relation between HbA1c and antioxidant markers along with lipid profile in these patients. Results: There was significant increase in fasting plasma glucose (FPG) (p<0.001), HbA1c (p<0.001), LDL-C (p<0.001) and ceruloplasmin (p<0.001), and there was significant decrease in serum thiols (p<0.001), GST (p<0.001), and HDLC (p<0.001) in type 2 DM patients compared to healthy controls. HbA1c correlated positively with FPG (r = 0.749, p<0.001), LDL-C (r = 0.513, p<0.001), and ceruloplasmin (r = 0.568, p<0.001) and negatively with HDL-C (r = -0.715, p<0.001), Methods: Blood samples were collected from thiols (r = -0.704, p<0.001) and GST (r = -0.426, healthy controls (n=55) and Type 2 DM patients p<0.001). (n=55) and their fasting lipid profile and HbA1c Conclusion: Findings in our study indicate the levels were determined using auto analyser. possibility of increased glycation of bio molecules Antioxidant status markers such as serum thiols, and enhanced generation of free radicals leading ceruloplasmin and GST are measured to deficient antioxidant status and altered lipid profile in favour of increased atherogenesis spectrophotometrically. Abstract No: 10.2

Lipid Indices in Type 2 Diabetes Mellitus and Their Association with Macro and Micro-vascular Complications

Dr. Imran Ahmed Siddiqui a , Dr. J Rama Rao b and Dr. Prabhavati Modi c a Postgraduate MD biochemistry, Department of Biochemistry Osmania Medical College, Hyd. b Former Professor & Head Department of Biochemistry, Osmania Medical College, Hyd. c Professor & Head Department of Biochemistry, Osmania Medical College, Hyd. Email: [email protected] Objective: To identify changes in lipid metabolism in Type II DM in context with the glycemic status, its relative impact on the macro & micro vascular events, and the effects of insulin therapy on the lipid indices. method, Triglyceride levels by GPO-PAP method and HDL-C by using the phosphotungstate precipitation and CHOD-POD method. The Cholesterol/ HDL-C ratio and Atherogenic Index (AI) [Logarithm (log) of TG/HDL-C] were then Methods: 158 Type II diabetics as cases and 30 calculated. subjects without any coincidental illness as Results: Atherogenic index and CHOL/HDL-C are controls were selected for the study. Total significantly higher in diabetics than in controls cholesterol was measured using the CHOD-POD and in uncontrolled diabetics than controlled

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diabetics. Both the indices were also found to be lowered in patients on treatment with insulin. The AI in patients with complications was also significantly higher than those without complications; However CHOL/HDL-C was not significantly different. Thus using the best cutoff values AI can be used as a better indicator for complications than using the ratio of CHOL/HDL-C. Conclusion: An imbalance between the LDL and HDL particles, where in, increase in the number Abstract No: 10.3

of small and dense LDL and decreased HDL levels leads to atherogenic dyslipidemia and increased incidence of plaque. Both lipid indices indicated dyslipidemia in diabetics and lower values were associated with glycemic control and insulin therapy. Hence AI can be considered as a diagnostic marker of increased cardiovascular risk in patients with Type II DM, and a guide for aggressive therapeutic approach.

Evaluation of Prothrombin Time and Activated Partial Thromboplastin Time in Type-2 Diabetes.

Dr. Itishri Jena and Prof. Prakash Ch. Mohapatra S.C.B. Medical College and Hospital, Cuttack, Orissa, India. Email id: [email protected] Objective: Diabetes Mellitus is the leading endocrine disorder of global interest that leads to thromboembolic phenomena in future. The pathogenesis of diabetic vascular disease is not clearly known. Haemorrheological as well as hypercoagulable states have been suggested to cause this complication. In order to asses the status of coagulation, we investigated prothrombin time (PT) and activated partial thromboplastin time (APTT) in diabetic patients and their relation with blood glucose, Glycated haemoglobin (HbA1c) and insulin resistance. Methods: A study was undertaken to evaluate these parameters in 60 diabetics and 30 normal controls attending the out and in-patient department of S.C.B. Medical College and Hospital, Cuttack, during the period from July 2008 to March 2009. Both PT and APTT were Abstract No: 10.4 measured by an automated blood coagulation analyzer, CA-50, manufactured by Sysmex. Results: The average age of cases was 51.55 years with average duration of disease 5.8 years. Both PT and APTT were observed to be shorter in diabetics than in controls; though within normal range. The fall is more marked in diabetics with complications (p<0.05). Both PT and APTT revealed association with hyperglycemia and insulin resistance. Conclusion: Our study revealed an association between shortened PT and APTT with hyperglycemia which may lead to thromboembolic complications in future. Identification of such factors may allow identifying that subset of diabetic patients at risk of thromboembolic complications and special preventive measures may be provided to them.

Effect of Trigonella Foenum Graecum and Insulin on Altered Membrane Functions in Alloxan Diabetic Rat Brains

Pardeep Kumar, Asia Taha, R.K. Kale and Najma Zaheer Baquer School of Life Sciences, Jawaharlal Nehru University, New Delhi, India, Email: [email protected] Objective: To study changes in monoamine oxidase (MAO) activity, lipid peroxidation and

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insulin levels, membrane fluidity and lipofuscin accumulation occurring in diabetic rat brain and

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to see whether these changes are restored to accumulation in total brains of rats. Activity of normal levels after administration of insulin and catecholamine degrading enzyme, MAO showed Trigonella foenum graecum powder (TSP). a significant increase in the synaptosomes membrane fractions of whole brain in diabetic Methods: Animals were made diabetic by animals. Present study also showed that insulin injecting 15mg/100g body weight alloxan, to administration and TSP to diabetic animals 24hours starved rats. Diabetic animals were given significantly decreased MAO activity, lipid Trigonella seed powder (5% w/w) mixed with their peroxidation, membrane fluidity and lipofuscin standard food for 21 days. MAO activity was accumulation in brains of rats. assayed in synaptosomal and supernatant fractions of the brain. Lipid peroxidation was measured in Conclusion: It can therefore be suggested that different brain regions by measuring formation of Trigonella foenum graecum (TSP)'s beneficial 4-Hydroxynonenal (4HNE) levels. The presence effects on metabolic parameters is due to their of lipofuscin was observed by fluorescence antioxidative, antilipofuscin and microscopy in different brain regions. Insulin antilipidperoxidative actions. The results of such studies will be useful for pharmacological levels were measured using ELISA. modification of the diabetic process, for Results: Present work revealed that alloxan development of new antidiabetic drugs and diabetes was associated with significant increases normalizing the metabolic syndrome. in MAO activity, lipid peroxidation and lipofuscin Abstract No: 10.5

Metabolic Syndrome in Type 2 Diabetes Mellitus

Priya.K.Dhas*, Janet**, Shantharam** and Dr. Rita Mary Aruna * *Department of Biochemistry, Penang International Dental College, Vinayaka Missions University, Salem ** Department of Biochemistry, Madurai Medical College, Madurai. [email protected] Objective: Insulin resistant patients are at an increased risk of developing metabolic syndrome, a major cause of heart disease and dyslipidemia. Metabolic syndrome is a cluster of metabolic risk factors that come together in a single individual. Hence in the present study the prevalence of metabolic syndrome among Type 2 diabetes mellitus by WHO criteria has been carried out. The risk factors are compared between patients with Type 2 diabetes mellitus with and without metabolic syndrome. Methods: 87 age matched Type 2 diabetic patients attending a diabetic clinic were included in this study. The biochemical parameters like FBS, PPBS, lipid profile were estimated using diagnostic kits. Blood pressure was measured and BMI was calculated. The prevalence of metabolic syndrome was carried out by classifying the parameters based on WHO criteria. Results: The prevalence of metabolic syndrome as per WHO criteria was 33% in Type 2 diabetic patients. A significant increase was seen in TGL, VLDL, TGL/HDL ratio, BP, BMI and a decrease was observed in HDL of Type 2 diabetic patients with metabolic syndrome when compared with the Type 2 diabetic patients without metabolic syndrome. Conclusion: The significant increase in TGL, TGL/HDL ratio, BP, BMI of diabetic patients with metabolic syndrome when compared to the Type2 diabetic patients without metabolic syndrome predicts that patients with Type 2 diabetes mellitus with metabolic syndrome are at a higher risk level for CHD.

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Abstract No: 10.6

Study of the Effects of HCN Exposure in Cassava Workers

Priya.K.Dhas, P.Chitra and Dr. Rita Mary Aruna Department of Biochemistry,Penang International Dental College, Vinayaka Missions University, Salem. [email protected] Objective: Hydrogen cyanide is one of the chemicals responsible for tissue hypoxia. Chronic exposure to HCN may cause neurological, respiratory, cardiovascular and thyroid defects. Onset of symptoms depends on dose and duration of exposure. Large scale of cassava processing could be disastrous due to discharge of hydrocyanic acid into the air. Hence the present study is taken to assess the thyroid dysfunction leading to impairment in energy metabolism. Methods: 25 cassava workers in Salem and age matched controls were taken up for this study. Abstract No: 10.7 Their clinical history was obtained and the following parameters are estimated using a fasting blood sample. Blood sugar, lipid profile, serum total protein, urea, creatinine and T3, T4, TSH are estimated using diagnostic kits. Results: Our study reveals a significant increase in triglycerides and TGL/HDL ratio which is a risk factor for cardiovascular disease. This triglyceride is predominantly endogenous and may be synthesized from sugars that cannot enter energy producing pathways. The results of T3, T4 and TSH will be discussed.

Prevalence of Obesity in Female Students Residing in Hostels in the University Campus of Manipal

Rosemol Jacob M, Yeshoda K, Elsa Maria, Uma Pallavi, Asha Kamath, Dr. Revathy P Shenoy and Dr. Anjali Rao Department of Biochemistry, Kasturba Medical College Manipal, India-576104, E mail: [email protected] Objective: The fact that women are most vulnerable to obesity and our observation that considerable number of females residing in Kasturba medical college (KMC) hostels are gaining weight made us take up this study. Although obesity is a prevalent problem in all parts of world, very few data are available regarding obesity among young Indian women. This study was conducted to explore the problem among the female hostel inmates and to identify different factors contributing to obesity. Methods: Survey approach was a nonexperimental study in which we investigated 266 female hostel inmates of age group 17-27. Their

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biometrical characters such as height, weight, body mass index (BMI), waist-hip ratio (WHR), waist circumference (WC) and food habits were studied. Data analysis was done using SPSS (version 14). Results: According to waist-hip ratio, 227(85.3%) out of 266 females were found to be overweight where as only 32(12%) females were found to be overweight when BMI was considered. Conclusion: Significant correlation was observed between the different biometric characters. Since WHR is a slightly stronger predictor of CVD risk than WC or BMI, the female inmates of KMC Hostels, Manipal are susceptible to disorders associated with obesity.

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Abstract No: 10.8

In Vitro Effect of Visfatin on Glucose Utilization by Visceral Adipose Tissue

Radhika G1, Magnus J2, Sasikala M1, Venkat Rao G, Pradeep Rabella and Nageshwar Reddy D 1 Asian Healthcare Foundation, 2Asian Institute of Gastroenterology; 6-3-661, Somajiguda, Hyderabad- 500 083. E-mail: [email protected] Background: Visfatin a novel adipokine earlier recognized as pre â cell colony enhancing factor (PBEF), predominantly expressed in visceral adipose tissue has distinct activities of central importance in cellular energetics and innate immunity. It is recently shown to have insulin mimetic effects, bind and activate insulin receptor through a different binding site other than insulin, thus stimulate glucose utilization in peripheral tissues as well as reduce release of glucose from hepatocytes. conditions. At required intervals glucose in the medium was estimated by GOD-POD method using UV-Visible spectrophotometer. Insulin receptor phosphorylation was estimated by ELISA method.

Results: Visfatin treatment significantly increased (P< 0.001) insulin stimulated glucose utilization and insulin receptor phosphorylation in visceral adipose tissue obtained from lean subjects with normal BMI without central obesity. However, increase in visfatin induced insulin stimulated Objective: Present work was intended to study glucose utilization was not observed in tissues the invitro effect of visfatin on insulin stimulated obtained from overweight and obese subjects under glucose utilization by visceral adipose tissue identical experimental conditions. explants obtained from lean, overweight and obese Conclusion: In conclusion, our result shows that human subjects. Results may have therapeutic visfatin enhances insulin stimulated glucose implication in metabolic syndrome. utilization in lean human subjects. The observed Methods: Visceral adipose tissue explants inability of visfatin to increase insulin stimulated incubated in DMEM (nutrient medium with glucose utilization in overweight as well as obese 1000mg/l glucose) with insulin were exposed to subjects is substantiated by the insulin resistant visfatin and glucose under optimal experimental nature of those subjects. Abstract No: 10.9

Salivary Cortisol Level in Depression Patients

Sukanya Shetty1, Suhas Pattar2, Satheesh Rao3 and Srinivas Bhat4 a Student, K.S.Hedge Medical Academy, Mangalore, 1 Professor & Head, Dept. Biochemistry; K.S.Hedge Medical Academy, Mangalore, a2Student, K.S.Hedge Medical Academy, Mangalore, 3 HOD, Dept. Psychiatry; K.S.Hedge Medical Academy, Mangalore, d Asst Prof, Dept. of Psychiatry; K.S.Hedge Medical Academy, Mangalore. Backgfound: Major depressive disorder is a state activities of daily living for long period of time of intense sadness, melancholia or despair that Major depression does not have any specific disrupts individual's social functioning and/or markers but recent studies have shown that in

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depressive patients there is an elevated level of Salivary Cortisol. It is said that about 50% of patients with major depression hypersecrete Cortisol, which may be secondary to HPA overdrive. AIM: To compare the level of salivary cortisol in depressed patients and normal people. Methods: All 40 subjects were given GHQ (General Health Questionnaire). The subjects who score less than 20 in the GHQ were selected as control group (N -20) and the subjects who score more than 20 and fulfill the ICD-10 criteria for depression were selected as active group (N- 20). Psychiatrist diagnosed depression and also assessed the Severity of depression by HAMD(Hamilton Depression Scale). Morning sample(9:00am-12:00pm) of Saliva was collected Abstract No: 10.10

in a sterile plastic tube and stored under 0 degree Celsius refrigeration till the sample was analyzed for cortisol level. Results: The Mean of the Salivary Cortisol in Depression Patients was 1.81+0.54 ug/dL. Mean of the Salivary Cortisol in Control was 0.71+0.21 ug/dL. The mean difference were statistically significant, Z- value was 3.431 and the difference was found to be statistically significant @p<0.001. Mean Salivary cortisol level was low in the group at the GHQ Scoring 20-30 1.6+0.29 and higher in the GHQ Scoring of 30 and above. This was not significant. Conclusion: The Salivary Cortisol level was found to be significantly higher in depressed patients as compared with the controls.

Atherogenic Index of Plasma and its Relationship with Fasting Blood Glucose in Diabetes Mellitus Patients.

Supriya M, Kavya R, Phani M.N, .Jeevan K Shetty and Mungli Prakash Department of Biochemistry, Kasturba Medical College, Manipal-576104 E-mail id: [email protected] Objective: Dyslipidemia is very common in patients with diabetes mellitus. Diabetic dyslipidemia is characterized with increase plasma triacyglycerol and decrease HDL concentration. In present study we measured atherogenic index of plasma (AIP) & its relationship with fasting blood glucose in diabetes mellitus patients. Methods: Fasting glucose, fasting lipid profile and AIP were estimated in 50 diabetes mellitus patients and 50 healthy controls. Lipid profile and fasting blood glucose was analyzed by auto analyzer. AIP was calculated using formula log [TG/HDL-C]. Results: There was significant increase in Fasting blood sugar, total cholesterol, triacyglycerol, LDL

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cholesterol and AIP observed in cases (p<0.001) compared to healthy controls. There was significant decrease in HDL cholesterol (p<0.001) in cases compared to controls. Significant positive correlation was observed between fasting glucose &AIP (p<0.05) in cases. Conclusion: According to previous studies, elevated AIP levels increases the risk for development of atherosclerosis. In our study we observed elevated AIP in diabetic patients which predisposes them for development of atherosclerosis. The positive correlation between fasting blood glucose and AIP indicates poorly control of diabetes mellitus has got enhanced risk for development of atherosclerosis.

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Abstract No: 10.11

Lipoprotein (a), Fibrinogen, C ­ Reactive Protein, Uric Acid and Microalbuminuria in Coronary Artery Disease and Diabetes Mellitus"

Suvarna.T. Jadhav*, Dr. A.V. Sontakke** Dr. B.M. Tiwale*** and Dr. Mrs. C.C. Khanwelkar** Department of Pharmacology, Krishna Institute of Medical Sciences, Karad; *Department of Biochemistry, B. V. U. Dental College and Hospital, Sangli; ** Department of Biochemistry, Krishna Institute of Medical Sciences, Karad; *** Department of Biochemistry, Dr. D.Y. Patil, Medical College, Kolhapur; E-mail ID ­ [email protected] Objectives: 1) To study biochemical parameters Results: Significant increased in the levels of Lp (a) (P < 0.001), Fibrinogen (P < 0.001), C ­ as cardiac markers, like, Reactive Protein (P < 0.001), Sr. Uric Acid (P < a. Serum Lipoprotein (a) 0.05) & microalbuminuria (P < 0.001) was observed in patients with CAD & DM as compared b. Plasma Fibrinogen to normal healthy controls. c. C ­ Reactive Protein Conclusion: The increased incidence of CAD in d. Serum Uric Acid patients with DM may be due to increased levels e. Microalbuminurea of Lp(a) and Fibrinogen in DM patients which may 2) To study these parameters in patients of CAD be responsible for modified blood coagulation mechanisms. Increased C ­ Reactive protein with DM. values may predict future cardiovascular disease, 3) To establish the correlation between CAD and atherosclerosis being an inflammatory disorder, DM with reference to above parameterts. Elevated Sr. Uric Acid has been found to be closely Methods: Following methods were used for associated with dyslipidemia, abesity, estimation. 1.Lipoproein(a) by latex turbidimetry; hypertension, diabetes, smoking & inflammation 2 Fibrinogen by DADE. Behring method; 3 C- which are well known risk factors of CAD. reactive protein by turbilatex method; 4 Serum Microalbuminuria is associated with wide spread uric acid by Trinder method; 5 Microalbuminuria abnormalities in the vasculature that may manifest by immunoturbimetric method. as altered vascular reactivity and endothelial dysfunction. Abstract No: 10.12

Is hsCRP an Independent Risk Factor in Type 2 Diabetes?

Dr.V.S. Kalaiselvi Associate professor, Dept. of Biochemistry, Sree Balaji Medical College & Hospital, Chrompet-chennai; Email id:- [email protected] Background: Diabetic patients are known to have hsCRP( high specific C-reactive protein) a a manifold risk for coronary heart disease and molecule produced in response to inflammation , other complications. In Type 2 Diabetes, elevated is found to be an independent risk factor.

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Objective: 1. To estimate the level of hsCRP in latex turbidimetry. Lipid profile, blood sugar & non diabetic and diabetic individuals. 2. To find others investigations were done by routine methods. any correlation between hsCRP levels and lipid Results: Significant rise in hs CRP levels is noticed profile in type 2 diabetes. in diabetic patients when compared to non diabetic Methods: 100 individuals (n=100) were included individuals. hs CRP levels are correlating well with and grouped as follows, Group 1- patients with serum TGL levels in type 2 diabetics. type 2 diabetes (n=50) Group 2- non diabetic individuals (n=50); hs CRP level estimated by Conclusion: Elevated hs CRP levels, in type 2 Diabetes may be considered as an independent risk factor.

Abstract No: 10.13

Oxidative Stress and Proteolytic Activity in Erythrocytes of Diabetic Patients

Dr. Varashree B S1 and Dr.P.Gopalakrishna Bhat2 1 Assistant Professor, Department of Biochemistry, 2Professor, Department of biochemistry, Kasturba medical college, Manipal- 576104; Email Address: [email protected] Objective: To evaluate oxidant stress status in diabetes mellitus and its association with proteolytic system in erythrocytes. The parameters assessed are erythrocyte malondialdehyde (MDA) levels and proteolytic enzyme activity in both diabetic individuals and controls. Methods: 51 non diabetic patients (mean age=54.47); 53 diabetic patients (mean age=52.92) were included in the study. The diabetic status was assessed by estimating the fasting blood sugar (FBS) using glucose oxidase method. Oxidant stress was measured by estimating erythrocyte MDA in terms of thiobarbituric acid reacting substance (TBARS). Proteolytic activity by the amount of amino group released by the erythrocyte lysate of the diabetic individual using phenylhydrazine treated hemoglobin as substrate. Results: Erythrocyte MDA was higher in diabetics (4.7±1.7nmoles/gHb) than the controls

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(3.3±2.2nmoles/gHb) [p=0.001]. Erythrocyte proteolytic activity was higher in cases (167.2±648 nmoles/gHb) than the controls (27.9±31.7 nmoles/ gHb). FBS did not correlate with the erythrocyte MDA in diabetic patients and erythrocyte proteolytic activity. No correlation between erythrocyte MDA and erythrocyte proteolytic activity in cases. Conclusion: The erythrocyte lipid peroxidation was increased in diabetic patients. The proteolytic activity in the erythrocyte lysates of diabetic patients was significantly increased. There was no correlation of proteolytic activity and the TBARS in erythrocytes of diabetic subjects. Diabetes is associated with a significant increase in TBARS which is an index of oxidant stress. Proteolytic enzymes degrade many oxidatively altered proteins preventing the accumulation of altered and damaged proteins in the cell.

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Abstract No: 10.14

Leptin Levels in Early Post Transplant Period: Can it be Used to Assess Changes in Body Composition ?

Victoria Job, Venkataramana Raju* and George T John* Clinical Biochemistry, * Nephrology, Christian medical college, Vellore Background: Weight gain is common after renal transplantation, influenced by improved appetite and a reversal of the uremic state. An increased centripetal obesity and muscle atrophy accompany the long term glucocorticoid requirements. Such changes in body composition are associated with an increased risk of cardiovascular complications, which is a major cause of morbidity and mortality in renal transplantation. 33.4±11.2, had a mean weight of 52±10.1, 56.8±9.3 and 57.7 ±9.6 kg at base line 3, 6 months (p<0.01).The mean body fat at base line, 3 & 6 months were 11.1±5.7, 13.9±5.7 and 14.5±6.2 kg (p <0.001). The fat increment in arms, legs and trunk were 37.4%, 31.6% and 36.6%. Total bone mineral content decreased significantly at 6 months. The mean Leptin levels 5.3±7.9, 6.0± 8.4 and 15.4± 17.4 ng/ml (p<0.05) and it positively correlated with total body and regional fat content Objective: To assess the changes in body (p<0.01). composition and Leptin levels in the early post transplant period. Conclusions: Post renal transplant patients gain significant amount of weight, mostly due to Methods: Consecutive renal allograft recipients accumulation of fat, especially around arms and were assessed prospectively at baseline, 3 and 6 trunk. There is overall decrease in bone mineral months post-transplant by anthropometry, DEXA content, predominantly in the and serum Leptin levels. spine.Anthropometric measurements of fat content are a reasonable substitute for DEXA. Leptin levels Results: 62 recipients (M: F=3:1), mean age correlate with body fat content. Abstract No: 10.15

Inhibition of Type I 5á-reductase

Vijaya Patil a, Debjani Dasgupta a, Grace Samual b, Shubhangi Mirapurkar b, and Krishnamohan b a.Department of Biochemistry, The Institute of Science, 15 Madam Cama Road, Mumbai - 400032, India.; b. Board of Radiation and Isotope Technology,Vashi complex, Navi Mumbai-400705 Objective: Aim of this study was to investigate the inhibition of 5á-reductase type I enzyme using extracts from Indian medicinal plants. This enzyme converts testosterone into metabolites such as dihydroxytestosterone and androstenedione. Type I 5á-reductase has been implicated in skin disorders such as acne, hirsuitism and male pattern baldness and its inhibition offers a potential treatment for these disorders. Methods: Plant extracts selected for this study were screened and selected based on their ability to inhibit acne-causing microorganisms. PC3 cell lines were used as a source of 5á-reductase type I enzyme and experiments were conducted at pH7

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(optimum pH of the enzyme). PC3 cells were incubated for varying time intervals with testosterone (1.25 µM) in steroid free medium in presence or absence of increasing doses of seven plant extracts. Dutasteride and Finasteride were used as standards. The rate of uptake of Testosterone was studied using radioimmunoassay. Results: It was found that extracts of Picrorhiza

kurroa, Vitex negundo and Embelia ribes were potent inhibitors of 5á-reductase type I enzyme compared to known inhibitors such as Dutasteride and Finasteride. Conclusion: The present study indicates that extracts from Picrorhiza kurroa, Vitex negundo and Embelia ribes may be used for treatment of skin disorders involving 5-á-reductase type I enzyme.

Abstract No: 10.16

Thyroid Function in Diabetic Subjects

Ranjani.P and Vijaya Srinivasan Department of Biochemistry, Sri Ramachandra Medical College and Research Institute, Sri Ramachandra University, Chennai. Objective: Diabetes mellitus and thyroid disease are the two common endocrinopathies seen in adult population. Insulin and thyroid hormone being intimately involved in cellular metabolism excess or deficit of either of these hormones could result in the functional derangements of the other. Several studies have documented the prevalence of the thyroid disease in women with prevalence rates ranging from 4 to 21%and the rate in men being 2.8 to 16%. However the studies on the association of thyroid disease and diabetes are scarce. Failure to recognize the presence of abnormal thyroid hormone levels in diabetes may be the primary cause of poor management often encountered in some treated diabetics. The present study was therefore undertaken to evaluate the thyroid function in diabetic subjects. Methods: The study was undertaken at Sri Ramachandra Medical College, Chennai. People in the age group of 30-60 years with diabetes mellitus were included for this study. The control group consisted of normal subjects in the age group of 30-60 years. The blood sample obtained

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from the patients was processed. The thyroid hormones T3, T4, TSH and FT4 were analyzed by using automated analyzer ADVIA Centaur system. Results: The T3 concentration was lower (p< 0.001) in diabetic patients compared to normal subjects. A significant negative correlation was observed between the fasting blood glucose and T3 levels (r = -0.41) in diabetic patients. The decreased T3 levels could probably be due to decreased peripheral conversion of T4 to T3, increased degradation or clearance of T3 or else increased T3 metabolism via alternative pathway other than deiodination. No significant variation in T4,TSH and FT4 were observed in diabetic subjects. Conclusion: The finding of the present study suggests that alteration of thyroid hormone is a characteristic feature in diabetic patients. The detection of abnormal thyroid hormone levels in the early stage of diabetes mellitus will help the patients to improve quality of life and reduce the morbidity rate. Thus evaluation of thyroid profile is essential in diabetic subjects.

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Abstract No: 10.17

Atypical Findings of Oral Glucose Tolerance Test (OGTT) Curve in Gestational Diabetes Mellitus (GDM)

Shivaraj Gowda, P B Desai, Vijayetha S Kagwad, Sindhu J Shetty and M B Ilakal. Department of Biochemistry, J. N. Medical College, Belgaum 590010, Karnataka Email: [email protected] Background: Gestational diabetes mellitus is defined as any glucose intolerance detected during pregnancy associated with the metabolic risk of type-2 diabetes mellitus and high risk of maternal and perinatal morbidity. Early diagnosis is very important to prevent the complications and OGTT is the preferred test to detect GDM. The objective of the study was to analyze the pattern of OGTT curve and to understand the response of Insulin to the glucose metabolism. Methods: A total of 96 pregnant women at high risk were undertaken for OGTT and test performed at 24-28 weeks of gestation using 100gms of glucose. The test results were plotted on a graph Abstract No: 10.18 and interpreted using Carpenter and Coustan criteria. Results: 15 women were diagnosed as GDM and 5 out of them show atypical patterns in OGTT curve. Similar such patterns were also seen in 5 normal subjects. Conclusion: The shape of the OGTT curve is related to glucose metabolism, â-cell function and insulin sensitivity. Pregnancy is a state of physiologic insulin resistance. Insulin secretion pattern is disturbed in patients with insulin resistance. The atypical findings of OGTT will be discussed in the scientific presentation

Insulin Level in Obese Patients with Obstructive Sleep Apnea Syndrome.

Yogendra Singh, Surya Prakash Bhatt, *Randeep Guleria, and Manjit Singh Dhillo Department of Medicine All India Institute of Medical Sciences New Delhi-10023

Background: Obstructive sleep apnea (OSA) is caused by either complete obstruction or by partial obstruction of the upper airway and is known to be associated with metabolic, cardiovascular and neuropsychological disorders. OSA may correlate with Insulin resistance. Indians are more prone to Insulin resistance. We studied the correlation between Insulin levels and OSA in obese Indian patients.

levels were measured by Radio Immuno assay (RIA). Subjects with Apnea hypopnea index (AHI) >10/h were considered as obese with OSA and subjects with AHI <5/h were considered as obese without OSA.

Results: In this study, 49 % had severe, 14.6 % moderate and 19.4 % mild OSA and 17 % had normal study. BMI was high in severe OSA as compared to normal subjects. Mean AHI levels Methods: We enrolled 41 obese patients with significantly correlated with Insulin levels in the suspected sleep disordered breathing. Informed consent was taken. Demographic data and blood patients with OSA (p = 0.03). The mean serum samples were collected. All patients underwent a insulin levels was high in patients with OSA but split night polysomnography and serum insulin not statistically significant when compared to

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normal controls (p= 0.268). Overall obese subjects had 40 % higher serum insulin levels. Conclusion: We found a significant correlation between AHI and Insulin levels in Indian patients Abstract No: 10.19

with OSA. A larger sample size is needed to validate these finding and evaluate the relationship between insulin levels and OSA

Study of Lipid Profile, Oxidative Stress and Enzymatic Antioxidant in Type-2 Diabetes Mellitus

Youb Raj Neupane *, Dr Sandeep Roy**, and Dr Z G Badade*** *P.G. Student, *** Professor & Head, Department of Biochemistry, **Associate Professor, Dept of Medicine, MGM Medical College, Kamothe, Navi-Mumbai, Pin-410 209 [email protected] Objective: A distinguishable feature of type-2 diabetes besides hyperglycemia and deranged lipid profile is an impaired insulin secretion, peripheral insulin resistance, obesity and biological events due to oxidative stress has become major health concern worldwide. Methods: The study was designed to find out correlation between lipid peroxidation, enzymatic antioxidant and lipid profile, to rule out oxidative stress in type-2 diabetes mellitus. Patients with type-2 diabetes were diagnosed on the basis of WHO criteria. The degree of lipid peroxidation was measured in terms of malondialdehyde (MDA); enzymatic antioxidant viz. superoxide dismutase (SOD) along with lipid profile to assess Abstract No: 10.20 the status of oxidative stress in type-2 diabetic patients in comparison to the normal control. Results: There was significant increase in the product of lipid peroxidation and lipid profile except HDL cholesterol and superoxide dismutase, which was found, decreased in comparison to the normal control. From present study, it is clearly evident that type-2 diabetic patients were more prone to oxidative stress. Conclusion: It is concluded that controlling of hyperglycemia will prevent alteration in lipid metabolism and degree of oxidative stress that definitely plays an important role in prognosis and prevents secondary complications in type-2 diabetes mellitus.

Assay of CRP and HbA1C in Smokers and Nonsmokers with Type 2 Diabetes Mellitus in a Segment of Indian Urban Population

Rajyalakshmi Amancherla and Pradeep Naik Objective: Elevated high-sensitivity C-reactive protein (hsCRP) levels have been shown to be associated with type 2 diabetes mellitus (T2DM). The aim of the study was to assess the association of hsCRP with type 2 diabetes and to determine its correlates with HbA1C in a segment of urban population from the twin cities of Hyderabad.

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Methods: HbA1C and hsCRP were measured in 545 urban subjects by cation exchange chromatography and ELISA respectively. Results: Our results showed that a significant increases in hsCRP levels were observed with a mean (± SD) values of 1.65 ± 0.02 mg/L in normal

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smokers when compared to normal non smokers who had mean (± SD) of 0.96 ± 0.02 mg/L. HbA1C levels were also significantly increased in smokers who showed 6.5 %± 0.1 whereas non smokers showed 5.5 % ± 0.1 value. In the case of diabetes group smokers showed significantly higher values of hs CRP and HbA1c than their non smoker counterparts. The diabetic smokers showed 3.3 ± 0.2 mg/L hs CRP whereas the diabetic non smokers showed 2.1 ± 0.3 mg/L hs CRP in the serum. Similarly the diabetic smokers Abstract No: 10.21

showed 9.2 % + 0.2 HbA1c whereas the diabetic non smokers showed 7.99 % + 0.3 HbA1c in the blood. All the subjects were free from hemoglobinopathies and any other blood disorders. We demonstrate the association of low-grade systemic inflammation, as indicated by elevated hsCRP levels, with T2DM in Indian population. Smoking seems to play a major role in the increase of systemic inflammation irrespective of blood glucose levels.

Association of ApoA5 1131 T>C Polymorphism with Triglyceride Levels in Indians

R.Lakshmy, HPS Sachdev, Meenakshi Sharma, SK Bhargava, GR Chandak and KS Reddy Cardiac Biochemistry, AIIMS, New Delhi Objective: ApoA5 gene occurs near the ApoA1/ C3/A4 cluster on human chromosome 11q23. Knocking out ApoA5 gene in mice increases triglycerides by a factor of four. A polymorphism in the non coding region located 5' to the ApoA5 gene at nucleotide 1131 has been shown to be associated with high triglyceride levels in humans. The polymorphism results from T to C substitution at 1131. AopoA5 -1131C has been reported in 78% of Caucasians, 33% of Chinese and 35% of Japanese population. Since Indians have a higher prevalence of metabolic syndrome characterized by high triglyceride and low HDL, we looked at the relative contribution of ApoA5 T1131C polymorphism on triglyceride levels. Methods: The study was conducted in 1300 adults residing in Delhi. Polymorphism at ApoA5 T1131C was assessed by ARMS PCR. The lipid levels as well as LDL and HDL sub fractions were determined in all the subjects. Results: The associations of ApoA5 1131 genotype with lipid variables were computed. The TT genotype was present in 66% of the population studied, 30.5% had the TC genotype and 3.5% had CC genotype with the minor allele (C) frequency of 18.6%. This is higher than the 7-8% frequency of minor allele of ApoA5 (1131) reported in Caucasians. Subjects with CC had significantly higher fasting and post prandial triglyceride levels as compared to the subjects with TT genotype (p<0.001). 25% of the subjects with TT genotype had triglyceride >150, 32% of the subjects with CT genotype had hypertriglyceridemia where as 45% of subjects with CC genotype had high triglycerides. None of the other variables studied showed any association with ApoA5 1131 polymorphism. Conclusion: In conclusion ApoA5 variation may be an important determinant of triglyceride levels in Indians.

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Abstract No: 10.22

Minerals Status in Thyroid Disorders (Hypo & Hyper Thyroidism)

B.Suneel,D.R.Nagendra, K.Ramalingam and J.N.Naidu Department of Bio-chemistry, Narayana Medical College, Nellore. Objective: The Study was under taken to evaluate the changes of minerals in Thyroid disorders (hypo & hyper). TSH, FT3 & FT4 as markers of hypo & hyper Thyroidism & mineral Status Ca+, P+, Mg+, Zn+ are parameters studied. Increased P levels are (2.01 ± 1.03mg/dl) Decreased Mg+ levels are (3.8 ± 1.91mg/dl) Increased Zn+ levels are (167.42 ± 120.7ug/dl) decreased. In Hyper-thyroidism compared to controls Ca+ levels are decreased (6.60 ± 2.78mg/ dl) , P levels are Increased (7.07 ± 3.05mg/dl), Methods: The studies includes 25 controls with Mg2+ levels are Decreased (1.8 ± 0.87mg/dl), Zn+ normal TSH, T3& T4 . 25 Thyroid disorders having levels are Increased (60 ± 30.86ug/dl). below 4 years in age group of 30 ­ 48 years both sex were included TSH, T3, T4 was estimated by Conclusion: In this study the hypothyroid patients using ELISA Ca was estimated OCPC method should increased Ca and Mg levels decreased, P ,Mg2+ GEDTA method, Zn+ Nitro PAPS , P and Zn levels in hyperthyroid patients should Molybdate method. Decreased Ca and Mg levels, increased P and Zn levels. Results: In hypo thyroid patient compared with controls Ca + levels are (12.9 ± 11.1mg/dl) Abstract No: 10.23

Postprandial Hyperglecaemia and Reduction in Serum Paraoxonase Activity in Type 2 Diabetes Mellitus Subject: an Alarm of Vascular Complication

Raj Narayan Gupta*, Neelima Singh, S.K. Singh and Preeti Dohare *P.G. Department of Life Sciences, IASCA, ITM Universe, Gwalior, Department of Biochemistry, G.R. Medical College, Gwalior (M.P.),: [email protected]) Objective: The present study aimed to investigate PON activity and Lipid profile along with Lipid per oxidation in different group of diabetic and non diabetic with and without macro vascular complication and age matched healthy control. Methods: Blood samples were obtained from patients admitted in ICU or attending to OPD of Medicine of J.A. Group Hospital attached G.R. Medical, Gwalior. The Blood sugar (PPBS), HbA1c were measured in all the samples.

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Results: Significant decrease in PON activity along with HDL-c in type 2 diabetic is observed with increased postprandial hyperglycemia and HbA1c. the reduction in PON activity is further confirmed by elevation of Pro-oxidant P-MDA. Therefore PON activity along with HDL-c could be considered as diagnostic marker for macro vascular complications in NIDDM subjects and reduction in PON activity along with HDL-c can be alarming condition for uncontrolled postprandial hyperglycemia.

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Abstract No: 10.24

Anti-oxidant Potential (AOP) of LDL in Post Menopausal Women Suffering from Type 2 Diabetes Mellitus.

Nivedita Singh, Neelima Singh, Sanjeev Singh and Vishal Bhargava Department of Biochemistry, G.R. Medical College, Gwalior Objective : Menopause occurs as the age advances in life of women (40-50 years). Post menopausal phase is associated with absence of estrogen known to be cardio protective and potent antioxidant hormone. A strong association of absence of estrogen and cardiovascular complication in post menopausal diabetic female has been reported. and 50 post menopausal diabetic female subjects. Results: It is concluded from study that in these subjects besides hyperglycemia, hypertryglyceridemia and hypercholesterolemia also exists. LDL and VLDL were increased (P<0.001) with decrease of HDL cholesterol (P<0.001).

Methods: The present study has been carried out Conclusion: Sensitivity for oxidation of LDL in in 50 age matched healthy female (40-50 years) these subjects were found increased and therefore antioxidant potential (AOP) of LDL was decreased. Abstract No: 10.25

Analysis of Multivariate Factors in Metabolic Syndrome - a Correlation Study

Vinayak S Bhat 1, Reshma Kumarchandra 1, and Vijay S. Bhat 2 1. Kasturba Medical College, Mangalore, 2. Manipal Hospital Bangalore Background: Metabolic syndrome is a combination of medical disorders that increase the risk of developing cardiovascular disease and diabetes. It is a multifactorial disorder encompassing abdominal obesity, altered glucose and lipid metabolism, insulin resistance, hypoadiponectinemia, elevated blood pressure and high level of CRP. For the present study, controls were on prescribed diet and exercise and subjects selected had most of the characteristics of metabolic syndrome. Objective: To estimate the parameters such as Adiponectin, CRP and HOMA score, FBG, lipid profile, Homocysteine, HbA1c & to make a correlative study of multivariate factors using Kruskal­Wallis one-way analysis of variance and analysis of variance (ANOVA) involved in this abnormal clinical condition and also to check whether diet and exercise have any effect on reducing the abnormal level of the different parameters involved in this disorder. Methods: Two categories of subjects were taken for the study, first category consisting of controls (10 members) and second category consisting of tests (61 members). Fasting insulin was estimated by electrochemiluminiscence method. HS-CRP by Immunonephelometric method, plasma Adiponectin was estimated by ELISA. FBG was by Hexokinase method. Cholesterol was by CHOD-POD method & Triglyceride by GPO-PAP method. Statistical analysis was done by using Kruskal ­ Wallis one way analysis of variance and by ANOVA. Results: Inverse relationship was observed between Adiponectin level and HOMA score, as well as with HS-CRP. A direct relationship between CRP & Homocysteine, HOMA score &HbA1c, history & Homocysteine, blood pressure & Homocysteine was observed. These values were statistically significant (p-value < 0.0001). Post exercise for 2 months, there was an increase in level of adiponectin in all the controls which also

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indirectly brought in corrections in rest of the risk factors. Conclusion: After all the investigation and data analysis in this work on metabolic syndrome, we observed altered levels of various parameters like, CRP, adiponectin, homocystiene, lipid profile, HOMA score(insulin resistance), blood pressure, fasting blood glucose etc. When we compared these parameters with the controls Abstract No: 10.26

parameters we observed a significant difference in the values of parameters. So based on the results of this study, we come to a conclusion that proper diet and regular exercise has an effect on bringing down the different parameters to normal level and the persons who are on the borderline and prone to develop the metabolic syndrome can reverse it back with proper diet and exercise.

Diabetes Risk Prediction by Fructosamine in Non diabetic First Degree Relatives

Hegde A, D'souza F, Kaveeshwar V, Jose A, Tasneem S, and Manjrekar AP Dept. of Biochemistry, Kasturba Medical College, Mangalore-575004, Karnataka. Email: [email protected] Objective: A positive family history of diabetes increases the chances of developing the disease manifold. The earliest diagnostic marker is plasma glucose value and long term follow up through glycated hemoglobin (HbA1c). Fructosamine (FA) depicting short term glycemic control has been seldom tried. Hence, clinical usefulness of fructosamine estimation and its comparison with established markers before the actual onset of the disease was the purport. Waist Circumference (WC) and Blood Pressure was recorded and compared between groups.

Results: Group III had significantly higher mean FPG (150.26mg/dl) & HbA1c (8.23 %) values. In addition significant elevation in salivary FA (202.05 mg/dl) was also noticed. Group II was associated with elevated serum FA levels (533.62 mg/dl), increased serum FA/ total protein (TP) ratio & larger WC. In Group II it was also seen that WC & BMI correlated significantly and positively Methods: 23 non-diabetic first degree relatives with FPG (0.613, 0.400 respectively), while WC of diabetics (Group II) were compared with 27 correlated importantly with systolic BP (0.526). healthy controls (Group I) and 23 diabetic people (Group III). Fasting plasma Glucose (FPG), Conclusion: Serum FA has less sensitivity in Glycosylated hemoglobin (HbA1c), serum depicting chronic hyperglycemia as was seen in Fructosamine and Total proteins were estimated case of diabetics. Monitoring serum FA along with in fasting blood samples. Simultaneously saliva measures of WC and SBP could be used to foresee was analyzed for fasting salivary glucose (SG), the development of Diabetes and Cardiovascular salivary fuctosamine and Total proteins. BMI, disease in high risk individuals. Abstract No: 10.27

A Study of CVD Risk Factors in Metabolic Syndrome.

Sajimon Thomas and T.Vijayakumar Department of Biochemistry and Physiology, Educare Institute of Dental Sciences,Malapuram, Kerala -675506, India. Objective: Metabolic Syndrome is one of the commonest risk factors for Cardiovascular

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diseases. The major characteristics of metabolic syndrome include insulin resistance, abdominal

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obesity, elevated blood pressure and lipid abnormalities.The present study was aimed at establishing the link between Metabolic syndrome and it's role as an atherogenic risk factor in an Indian perspective. It is now established that this syndrome predicts the development of type 2 diabetes mellitus and cardiovascular disease.[1]The prevalence of the metabolic syndrome depends on age, ethnic background, and gender. It rises linearly from 20 to 50 years and plateaus thereafter. Looking at various studies around the world, which included population samples, aged from 20 to 25 and upwards, the prevalence varies from 8% (India) to 24% (United States) in men and from 7% (France) to 46% (India) in women.1-3

fibrinogen, High sensitivity C reactive protein( hsCRP) and Insulin were analyzed in both Case Group and Control Group Results: The two sample t statistical analysis of the results of the data from 225 individuals showed a statistically significant increase in all parameters in the Case Group compared to the Control Group (p<0.001). Conclusion: This establishes a strong link between Metabolic Syndrome and atherogenic risk factors substantiating similar observations elsewhere. References:

1. Vega GL. Obesity, the metabolic syndrome, and Cardiovascular disease. Am Heart J 2001;142:1108-16. Methods: The study was carried out in 225 individuals and the selection was primarily based 2. Ramachandran A, Snehalatha C, Satyavani K, on age, sex and socioeconomic status. 85 men and Sivasankari S, Vijay V. Metabolic syndrome in 65 women in the age group of 30 to 50 years and urban Asian Indian adults-a population study who are having 3 or more of the classified using modified ATP III criteria. Diabetes Res manifestations of Metabolic Syndrome were Clin Pract 2003;60:199-204. included in the Case Group45 men and 30 women in the age group of 30 to 50 years and who had 3. Deepa R, Shantiram CS, Premlalitha G, Shanti NG, Mohan V. Prevalence of insulin resistance none of the above mentioned manifestations syndrome in a selected south Indian populationcomprised the Control Group. Blood/Serum the Chennai urban population study-7 [CUPS- 7]. Glucose, Total cholesterol, HDL cholesterol, LDL Indian J Med Res 2002;115:118-27. cholesterol, Triglycerides, Uricacid Plasma Abstract No: 10.28

CVD Risk Factors in Women with Subclinical Autoimmune Thyroiditis.

Sudheesh.M and T.Vijayakumar Department of Biochemistry and Physiology, Educare Institute of Dental Sciences,Malapuram, Kerala -675506, India. Objective: Sub clinical hypothyroidism (SH), defined as the clinical status of mildly elevated serum TSH levels, is a far more common disorder than overt hypothyroidism with a higher prevalence among women and older subjects. Certain studies have indicated that sub clinical hypothyroidism has been associated with increased risk of CAD, especially in women with antibodies to thyroid peroxidase. Moreover, SH impairs ventricular function as well as cardiovascular and respiratory adaptation to effort and, decreases heart rate variability impairs flowmediated vasodilatation, which is a marker of endothelial function 1-3. The present study is an effort to investigate the CVD risk profile in women having autoimmune sub clinical hypothyroidism with elevated Anti Thyroid Peroxidase Antibodies.(Anti TPO Ab.)

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Methods: The study was carried out in blood samples from 150 women in the age group of 35 to 50 who are suspected to have hypothyroidism were included in the Case Group.80 apparently healthy women in the age group of 35 to 50 with normal serum TSH levels comprised the Control Group. Blood / serum TSH, Free T3, Free T4, Anti TPO Ab, Total Cholesterol, LDL cholesterol, Triglycerides , Plasma fibrinogen and High sensitivity C reactive protein ( hs-CRP) were analyzed in both Case and Control Groups. The data from the blood investigations were analyzed statistically using Two Sample`t' test. Results: All the blood / serum parameters which were analyzed in our study were found to be increased significantly in the Case Group when compared to the Control Group (p < 0.001). Conclusion: Thus the Case Control study in 200 women having sub clinical hypothyroidism with Abstract No: 11.1

elevated Anti Thyroid Peroxidase Antibodies showed that they are at a higher risk for CVD than their age-matched control counter parts. References 1. Ross DS: Serum thyroid ­ stimulating hormone measurement for assessment of thyroid function and disease. Endocrinol Metab Clin North Am 2001, 30:245-264. 2. Walsh JP, Bremner AP, Bulsara MK, et al. Subclinical thyroid dysfunction as a risk factor for cardiovascular disease. Arch Intern Med 2005; 165:2467-2472. 3. Hak AE, Pols HA, Visser TJ, Drexhage HA, Hofman A, Witteman JC. Subclinical hypothyroidism is an independent risk factor for atherosclerosis and myocardial infarction in elderly women: the Rotterdam Study. Ann Intern Med. 2000;132:270­278.

Recurrent Aphhous Ulcer and Oral Lichen Planus: a Study of Patho-physiological and Psychosomatic Significance of Salivary Nitric Oxide.

AB Agte* Bm Despande* Sm Dharwadkar** Kiran Jagtap* Sb Gaikwad*** and S.R. Holkar****, *SBM Dental College, Sangamner, Ahmednagar, **SB Science College, Aurangabad, *** SBH; Government Medical College, Dhule, **** MGM Medical College, Aurangabad. Objective: The aim of the present study was, to study the role of Nitric Oxide in human saliva & its diagnostic role in RAU & OLP, at outpatient in SBM. Dental College & Hospital, Sangamner, District Ahmednagar. Twenty (20) cases with RAU, Fifteen (15) with OLP & Thirty (30) healthy control individuals were included in the study. The clinically diagnosed known cases of RAU & OLP were included after taking detail case history. The immunological imbalance & psychological stress are considered to be the most prime factors for the conditions like RAU & OLP. Further; the Oxidative stress caused by Nitric Oxide has a definite role in establishing the correlation between [NO] with RAU & OLP.

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Methods:: Under all aseptic precautions, fresh saliva was collected in sterile plain bulb. The sample was subjected to biochemical analysis of Nitric Oxide (NO) by diazotization reaction with adoption of the principles of Griess reagent. The RAU cases were subdivided into major & minor RAU & OLP cases were divided into erosive or non erosive type after clinical & histological examination. Results: The salivary Nitric Oxide levels were found to be increased significantly In RAU & OLP group when compare with Control. Further significantly increased levels have been observed in OLP group when compared with RAU group

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(P< 0.001). The Salivary Nitric Oxide levels were Conclusion: Thus these parameters can be treated found to be increased significantly in minor RAU as a diagnostic tool for the differential diagnosis than Major RAU & increase in erosive type of of RAU & OLP. OLP than non erosive OLP. Abstract No: 11.2

Oxidative Stress and Calcium Phosphorus Ratio in Rheumatoid Arthritis

Dr.A.Pullaiah*, and DR.N.Vani *P.G.in Biochemistry; ** Professor&HOD, Dept of Biochemistry, Kakatiya Medical College, Warangal-506007. Andhrapradesh. Background: Rheumatoid arthritis is a common debilitating, crippling disease with unknown etiology associated with chronic inflammation and bone resorption. Increased oxidative stress contributes to the etiopathogenesis of R.A. Objectives: Present study aims to determine the oxidative injury by estimating the levels of prooxidant Malondialdehyde and to study the role of oxygen derived free radicals in R.A by assessing the influence of disease process on serum calcium, phosphorus, ALP levels in a view to study the beneficial influence of adding free radical scavengers to the existing therapeutic regimen. Methods: The study was conducted on 50 clinically diagnosed cases of R.A who fulfill the ARA criteria, compared with age & sex matched controls with out systemic diseases like DM, HTN, RF. Estimation of Malondialdehyde was done by TBA assay, Calcium by O-CPC method, Abstract No: 11.3 Phosphorus by Fiske and Subbarao method, ALP by King & Kind. Statistical analysis was done by using student "t" test and p-value <0.05 were considered as statistically significant. Results: The results that were found are statistically significant increase in serum MDA levels (p<0.001), decrease in serum Calcium (p<0.001), increase in serum Phosphorus (p<0.01), decreased Ca,P ratio (p<0.001) and with no statistically significant change in serum ALP levels (p>0.05) in R.A patients as compared with control group. Conclusion: The observations in the present study shows that oxidative stress plays an important role in the etiopathogenesis of R.A and addition of free radical scavengers as supplements may prove to be beneficial. There is an altered Calcium and Phosphorus metabolism and Ca,P ratio in R.A and increase in MDA levels, an important marker.

The Role of Oxidative Stress and Fibrinogen Level in Chronic in Diabetes Mellitus

Mrs. G. Ramani, Dr. Rita Mary Aruna, Dr. T. V. Mohan, and Mrs. G. Kavitha VMKV Medical College, Salem. Objective: Diabetes mellitus is a commonest endocrine disorder which represents the serious risk factor for the development of cardio vascular diseases. Oxidative stress is a pathophysiological state in which there is excess of Reactive Oxygen Species. In diabetes mellitus oxidative stress results from hyperglycemia. Oxidative stress alters the plasma lipoprotein profile, the coagulative parameters, the endothelium and the cell membrane. Hence the present study was focused to understand the total antioxidant level by Ferric Reducing Antioxidant Antioxidant Power assay. Methods: To conduct this study, blood samples were collected from the thirty patients from the Department of Medicine, Vinayaka Mission Medical College, Salem. The level of total

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antioxidant status was measured. The Lipid profile development of vascular disease. A negative was studied along with the level of Fibrinogen. correlation by Pearson has been observed between The results were compared with a control group. FBS and Ferric Reducing Antioxidant Antioxidant Results: The present study has demonstrated a Power as well as HDL and Ferric Reducing significant increase in FBS, PPBS, Total Antioxidant Antioxidant Power. cholesterol, Triacylglycerols, very low density cholesterol, low density cholesterol & Fibrinogen levels and decrease in Total antioxidant level and HDL in chronic diabetes mellitus patients. This appears to be related to increased production of lipid derived free radicals. Thus FBS oxidative stress seems to play an important role in the Abstract No: 11.4 Conclusion: This is suggestive of increased FBS as well as decreased HDL cause oxidative stress which may lead to atherogenesis. Therefore, the screening of diabetic patients in the initial stage for abnormalities in lipid metabolism and oxidative stress will be beneficial in the prevention of diabetic vascular complications

The Effect of Yoga Therapy on Antioxidant Level in Perimenopausal Women

1

Alok Kumar, 1Archana E.,1 Akshatha, 2Mrs. Gayathry Nayak, 1Dr.Revathi P. Shenoy and 1Dr. Anjali Rao 1 Dept. Of Biochemistry, Kasturba Medical College, Manipal, India, 2Dept. Of Yoga Therapy, Kasturba Medical College Hospital, Manipal, India; Email : [email protected] Objective:: To assess the efficacy of yoga therapy on antioxidant levels of perimenopausal women. Methods: It is a prospective, non-randomized control study. Consecutive sampling method was used for the selection of perimenopausal women aged between 40-60 years. There were yoga (n=30) and control (n=30) groups. The yoga intervention was on daily basis for 12 weeks (4550- minutes/day) and the control group intervention was a set of standardised physical exercises (15 minutes/day) and walking (20-25 minutes/day). The total antioxidant (TA) levels in serum were measured spectrophotometrically. Results: Total antioxidant levels are increased significantly (p<0.025) after three months of exercise and decreased significantly (p<0.048) after three months of yoga. There is a significant decrease in TA in post yoga group (p<0.001) as compared to post exercise group (control). Conclusion: There is a significant decrease in total antioxidant levels after yoga intervention probably due to increased consumption of TA to prevent the oxidative stress when compared to exercise group (control).

Abstract No: 11.5

Aluminum Induced Morphological and Biochemical Changes in Liver, Kidney and Brain of Male Albino Rats; An Age Dependent Study

Anumesh Pathak1,2, Sandeep Tripathi1, A. K. Pathak2, Abbas Ali Mahdi1 , Sanjay Sen1,2 , Ramkrishan Ojha1,2 and Neelam Dubey1,2 1 Medical Elementology and Free Radical biology lab, Department of Biochemistry, C.S.M. Medical University, Lucknow, 226003,2Department of Pharmacy, Barkatullah University, Bhopal, India Objective: Substantial evidence supports the hypothesis that aluminum (Al) is toxic for human

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beings. To date, information on the interaction of Al with oxidative stress parameters in the rat

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organs in relation to age was limited and unclear. old rats but the retention was high in old than the young rats. Oral administration of AlC3 increases Methods: The purpose of the present study was Al, Fe, LPO, and GSSG and decreases SOD, CAT, to investigate the effects of Al-induced toxicity GPx, GSH, and GSH/GSSG ratio, significantly (100 mg/ kg b.w. AlCl3 orally for 90 days) on age (p<0.01) in comparison to control. No significant (young and old) and organs (liver, kidney and (p>0.05) difference were observed in the activity brain) of male albino rats. For this, selected of CAT and GPx and level of LPO, GSH, GSSG oxidative stress biomarkers- lipid peroxidase and ratio of GSH/GSSG in the organs of young (LPO), superoxide dismutase (SOD), catalase treated and old untreated rats suggests treatment (CAT), glutathione peroxidase (GPx), reduced made young equivalent to old. Histopathological glutathione (GSH) and oxidized glutathione and ultrastructural study also correlated with (GSSG) along with metals- Al and Fe were biochemical findings. investigated in the brain, kidney and liver of male albino Wister rats. In addition, morphological Conclusion: These findings suggest that oral (Histopathological and ultrastructure study) and administration of AlCl3 may produce pro-oxidant Y maze test were also evaluated. effect in rats and could be of interest for understanding the controversial role of Al in Results: Results show that the oral administration assessing toxicity and neurodegenerative diseases of AlCl3 produce significantly (p<0.01) oxidative in exposed young and elderly population. damage in liver, kidney and brain of young and Abstract No: 11.6

Reduced Nitric Oxide Synthesis in Pre-eclampsia: Role of Inflammatory Cytokines and eNOS Gene Polymorphism

Archana Singh. 1, Deepika Sharma.1, Chitra Raghunandan.2 and Jayashree Bhattacharjee.1 1-Deptt. of Biochemistry, 2- Deptt. Of Obstetrics and Gynecology, Lady Hardinge Medical College, New Delhi, India.Email: [email protected] Results: Pre-eclamptic group showed significantly decreased serum NO levels whereas TNFá, IL-6, IL-2and ET-1 were increased significantly. No significant differences were found in genotype/allele distribution among the two groups. A significant negative correlation was seen between NO and IL-6 in pre-eclamptic group. Levels of NO were found to be decreased in women with GT Methods: This cross-sectional study included 50 genotype whereas levels of IL-6 were found to be pregnant women diagnosed with Pre-eclampsia increased in the same. and 50 healthy pregnant women. Their blood samples were analyzed for NO, TNF á , IL -6 , Conclusion: Pre-eclampsia occurs when there is IL-2 and Endothelin-1 and eNOS gene polymor- failure of compensatory response in terms of dephism. creased vasodilatation. NO, a potent vasodilator

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Background: Pre-eclampsia is associated with nitric oxide dysfunction. In order to gain more insight into the mechanisms underlying pre-eclampsia, we evaluated the effect of endothelial nitric oxide synthase Glu298Asp gene polymorphism and inflammatory cytokines on levels of serum NO in pre-eclamptic pregnant women.

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is decreased in pre-eclampsia. Increased IL-6, contributes to increased vascular resistance by inhibiting endothelium dependent NO-cyclic guanine monophosphate mediated relaxation pathway in systemic vessels in pre-eclampsia. In the present study no significant association was found beAbstract No: 11.7

tween Glu298Asp polymorphism and pre-eclampsia. However, this polymorphism decreases NO levels, and women with this polymorphism might develop CAD later in life. Much larger studies are needed to confirm or refute a realistic genotypic risk of disease.

Liver Function and Total Antioxidant Capacity in Hypothyroidism

Bhawna Bhimte*, Dr.B.K.Agrawal*, Dr.V.K.Sharma** and Prashant Nigam* *Department of Medical Biochemistry, Gandhi Medical College, Bhopal, **Department of Medicine, Gandhi Medical College, Bhopal . Email: [email protected] Objective: Thyroid hormone plays important role in normal hepatic function. Hypothyroidism may perturb normal metabolic function of liver. Low activity of thyroid hormones may also be associated with oxidative stress causing decrease antioxidant capacity. Methods: Prospective study has been done to assess liver function and total antioxidant capacity in 100 hypothyroid patients compared to healthy controls. Following parameters has been measured at the time of detection & then after 6 weeks. TSH: Competitive Solid Phase Enzyme Immunoassay Alkaline Phosphatase: p-nitrophenyl phosphate Gamma Glutamyl Transferase: End point pnitroaniline method Total Antioxidant Capacity: Korvacevic method. Results: TSH level of hypothyroid patients were Abstract No: 11.8 found to be increased significantly at the time of detection when compared to controls (P<0.001), but values were found to be reduced after 6 weeks of therapy. Activity of both liver enzymes ALP, GGT were found to be reduced in hypothyroid patients at the time of detection, but after 6weeks of therapy values of ALP & GGT were improved. (P<0.05). Total Antioxidant Capacity were also reduced in hypothyroid patients, that was found to be improved after six weeks of therapy (P<0.001). Conclusion: From above results it may be concluded that hypothyroidism modulate normal liver function, thus interfering with the liver enzyme production or probably its activity. Increased TSH may stimulate excess oxidative stress, thus creating load on Antioxidant Capacity.

A Study on Free Radical Activity in Burns

Dr. Bikash Kr. Chaudhury and Dr. Runi Devi Gauhati Medical College & Hospital, Guwahati-781032, NEIGRIHMS, Shillong-793018 Email: - [email protected] Objectives: The metabolic response to thermal sustained injury is mediated not by the burns itself, injury is similar to the metabolic response to stress but by the ROS generated secondarily. The study following trauma and about two-thirds of the was undertaken assess free radical activity in burn

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cases, to correlate free radical activity with extent Results: In the test subjects serum MDA and serum and depth of burn injury and to utilize data for Uric acid was significantly raised whereas serum better management of burn patients. TIBC, blood Glutathione Peroxidase and Superoxide dismutase levels were significantly Methods: 35 cases of burn injuries admitted to depleted compared to the controls establishing the Gauhati Medical College and Hospital, Guwahati relation that oxidative stress is associated with and 35 healthy individuals, age and gender increased burn trauma severity resulting in matched for the patients were studied. antioxidant depletion, strong oxidant production Malondialdehyde, Superoxide Dismutase, and cellular damage. Glutathione Peroxidase, Uric acid and Total Iron Binding Capacity (TIBC) were using a Conclusion: The study demonstrates the colorimeter, a semi automated analyzer relationship between a lipid peroxide reaction and (Humanlyser 2000) and the fully automated Dade secondary pathological changes following burns. Behring AR series. Statistical analysis was done Therefore early introduction of prophylactic AnalystSoft, StatPlus statistical analysis program antioxidant therapy may be beneficial following Version 2006. The results were taken as significant trauma and is highly recommended. when the `p' value < 0.05. Abstract No: 11.9

Oxidative Stress in N-acetylcysteine Treated Off Pump CABG Surgery Patient

Shyama Subramaniam, S.Subramaniam, K.Dhanajayan, K.Ezhilarasan, K.Baskar* and B.Jalakandan* Department of Biochemistry, Apollo Hospitals, Chennai ­ 600 006. *Department of Anesthesiology, Apollo Hospitals, Chennai ­ 600 006. Objective: To compare the effect of Nacetylcysteine (NAC) on off-pump CABG induced oxidative stress by analyzing the plasma malonedialdehyde (MDA) and glutathione levels. Results: A marked reduction of GSH and significant elevation of MDA were noticed after surgery in off pump CABG patients (P<0.001) . Pre operative treatment of NAC increased the GSH level and reduces the MDA level.

Methods: Plasma malonedialdehyde (MDA) and glutathione (GSH) were analysed in blood samples Conclusion: Off pump CABG operation increases obtained from 50 patients undergoing CABG, pre- the oxidative stress but NAC treatment reduces the operatively and after surgery. The patients were oxidative stress by increasing GSH level. divided into two groups with and with out NAC treated patients. Abstract No: 11.10

Estimation of Lipid Profile, Vitamin E and Malondialdehyde, in Pre-eclampsia

*Dr Chandan Kumar Nath and Dr Upasana Baruah *Senior Resident, Department of Biochemistry, NEIGRIHMS, Shillong-18; E [email protected], Objective: The present study was undertaken to evaluate the metabolic correlation between

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preeclampsia and oxidative stress. Methods: We selected 95 no of subjects for the study out of which normal nonpregnant control group had 25 subjects, normal pregnant control group had 40 subjects & preeclampsia group constituted of 30 subjects. The investigations included are serum lipid profile consisting of serum total cholesterol, serum triglycerides, VLDL, LDL and HDL, serum vitamin E as á- tocopherol and malondialdehyde. Results: There is significant increase in serum triglyceride, serum cholesterol along with LDL and VLDL (p<0.0001) in pregnancy over nonpregnant controls and preeclampsia over pregnant control (p<0.0001). It is observed that in normal controls without pregnancy there is no relationship between Abstract No: 11.11

serum vitamin E and serum lipid peroxidation as represented by whole blood MDA (malondialdehyde) with a correlation coefficient of 0.13 only. In contrast to this, under the condition of normal pregnancy and preeclampsia the correlation between these two parameters increased with coefficients of correlation (r = - 0.9) with a negative trend indicating a definite and significant degree of inverse relationship between two. Conclusion: It is finally proposed that adequate vitamin E supplementation during pregnancy with proportionate increase in preeclampsia may reduce the consequences of peroxidation induced complications during pregnancy.

Antioxidant Status in Goats During Pregnancy

Cynthia Jose* and K.K. Jayavardhanan** * MVSc Scholar, ** Associate Professor, Department of Veterinary Biochemistry, College of Veterinary and Animal Sciences, Mannuthy, Thrissur- Pin 680651 [email protected] Objective: Production of free radicals is a physiological phenomenon which occurs continuously in the body during metabolism and body usually has sufficient antioxidant reserves to cope with its production. However, when free radical generation exceeds the body's anti-oxidant production capacity, oxidative stress develops. The aim of the present study was to evaluate under field condition the oxidative status in healthy goats during pregnancy on the basis of 3 parameters: plasma levels of malondialdehyde (MDA), reduced glutathione (GSH) and glutathione peroxidase (GSH-px). In order to evaluate metabolic homeostasis, common metabolic parameters (glucose, total protein, albumin, cholesterol, urea and creatinine) were determined as well. Methods: The study was carried out using 15 healthy female cross bred goats maintained at

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Kerala Agricultural University Goat Farm, Mannuthy. Blood samples were collected 4 times: before pregnancy, at 2 nd and 4 th months of pregnancy and one week before kidding. The values obtained during pregnancy were compared with those obtained in the same animals before pregnancy. Results: The study showed that the lipid peroxidation was increased in the pregnancy period as revealed by an increase in MDA level. Significant higher MDA level was observed in the 4th month of pregnancy. This suggested that after the middle gestation period, the body presents high levels of free radicals which cause lipid peroxidation. The activity of anti-oxidant enzyme, GSH-px and the level of GSH were also found increased in this period, providing protection to the fetus against the negative influence of free radicals. The serum glucose concentration was

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significantly lower in the 4th month of pregnancy, indicating low energy balance in this period. The cholesterol and creatinine concentrations were higher in pregnant goats with a peak level in the late pregnancy period. Abstract No: 11.12

Conclusion: The data suggest that the antioxidative/prooxidative status is associated with the metabolic adaptations to low energy balance, which occur during pregnancy.

Comparative Study of Lipid Peroxidation and Non-enzymatic Antioxidant Vitamin E and Vitamin C in Plasmodium Vivax and Falciparum Malaria in Western Rajasthan with special reference to Bikaner District.

R.K. Vyas, D. Yadav, M.L. Sharma, Y. Soni and D. Nandini Department of Biochemistry S.P. Medical College, Bikaner (Rajasthan) Objectives: - Oxidative stress and changes in antioxidant status have been implicated in the pathogenesis of malaria. Bikaner district, being a part of Thar desert due to extremes of temperature has always been regarded as hypoendemic region for malaria but in recent years prevalence of Malaria has increased in this region due to changes in Ecosystem contributed by increased rainfall and Canal Irrigation. So this study was conducted in patients with Plasmodium falciparum and Plasmodium vivax malaria to evaluate status of lipid peroxidation and effect on vitamin E and C level. Methods: - In study we included 100patients out of which 50were suffering from Plasmodium vivex Malaria and other 50with Plasmodium falciparum Malaria. Plasma Thiobarbituric acid reactive substances(TBARS) were measured using Buege & Aust's Method(1978) to assess the degree of lipid peroxidation and antioxidant status was measured by estimating the level of Vit E&C Abstract No: 11.13 using Baker & Frank's Method(1976) using Bipyridyl reagent and Roe & Kuethe's Method using 2.4DNPH respectively. Results were compared with age and sex matched control subjects(n=50). Results: Our study demonstrates that level of oxidative stress was significantly increased (P<.001) while antioxidant vitamins E&C were significantly decreased (P<.001) in both the groups of malaria. Plasmodium falciparum (2.82+ 1.06 nmol/ ml) malaria showed significantly increased (P<.001) level of Malondialdehyde when compared to Plasmodium vivax malaria (1.72+ 0.70nmol/ml). Maximum decline in Vit C level (p<.001) were observed in plasmodium vivax malaria while maximum decline in Vit E level (p<.001) were observed in Plasmodium falciparum malaria. Therefore it has been hypothesized that the oxidative stress induced by malaria can be, combated by augmentation of Vit E&C.

Comparative Study of Serum Myeloperoxidase Level and Lipid Profile in Healthy Individuals

Suchetha Kumari. N* and Ramitha K** * Professor, Dept. of Biochemistry, K.S Hedge Medical Academy, Deralakatte, Mangalore -575018, ** Research Assistant, Central Research Lab, A.B.S.M.I.D.S Deralakatte, Mangalore -575018, Email: [email protected] Background : Myeloperoxidase (MPO) is an ironcontaining heme protein, plays an important role

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in host defense. It is one of the enzymes of the innate immune system, which acts as a potential marker of cardiovascular disease and potential target for treatment. Myeloperoxidase oxidizes LDL rendering it atherogenic, as well as oxidative modification of apolipoprotein. Objective: The present study was designed to compare the serum myeloperoxidase level and lipid profile in healthy individuals. Methods: This study was carried out at K.S.Hegde Medical College Hospital. Fifty volunteers of age between 30-40 yrs were selected for the study. About 5 ml of overnight fasting blood was collected and used for the estimation of lipid profile and Myeloperoxidase levels by Abstract No: 11.14

spectrophotometric method. The subjects were grouped into low lipid profile subjects and high lipid profile subjects based on their cholesterol level. The data were analyzed for statistical significance by one way ANOVA and P<0.05 was considered as the level of significance. Results: Myeloperoxidase levels were positively correlated with cholesterol, LDL, and Triglycerol levels in both high and low lipid profile subjects. But, HDL has shown a negative correlation with the Myeloperoxidase level. Conclusion: In this study we observed that individuals with high lipid profiles showed an increased Myeloperoxidase activity, which might be due to peroxide ions of lipids.

Nitric Oxide and Lipid Profile in Type 2 Diabetes Mellitus-- A Case Control Study in East Sikkim

Dr A Ghosh, Ms Y Bhutia, Dr T A Sing and Dr M L Sherpa Department of Biochemistry, Sikkim Manipal Institute of Medical Sciences, 5th Mile Tadong, Sikkim737102; Email- [email protected] Objective: Diabetes Mellitus has reached epidemic proportions with an alarming rise in the Asian countries including India. This particular study has been taken up to compare the serum NO level in patients with diabetes and healthy controls and to establish correlation between serum nitric oxide level and diabetes mellitus in Sikkim. Methods: Lipid Profile and serum nitrite were assayed in 52 subjects, 26 of which were diagnosed cases of type II Diabetes Mellitus undergoing treatment in SMIMS and 26 were healthy volunteers. The statistical analysis was done using SPSS 17 and the comparison between cases and control was calculated by Student's t ­test.

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Results: Most of the patients were above 40 years of age. Mean serum nitrite was lower in diabetics in comparison to non-diabetics but the difference was insignificant (p=0.141). Triacylglyerol, LDL were markedly raised in Diabetics but statistically insignificant (p=0.08), (p=0.55). HDL was significantly lower in Diabetics in comparison to non diabetics (p=0.001). Total Cholesterol was markedly lower in diabetics but this difference was statistically insignificant. Conclusion: The pattern of findings in our studies is consistent with the findings elsewhere but not statistically significant. This could be due to the duration of diabetes and the role of drugs, dietary and lifestyle modification prior to the study.

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Abstract No: 11.15

MDA, Antioxidant Enzymes, their Correlation in Normotensive and Pre-eclamptic Maternal and Cord Blood

Mohd Suhail1, Safia Suhail2 , Bharat Kumar Gupta3 and Vinay Bharat4 1. Department of Biochemistry, University of Allahabad, Allahabad-211002, India., 2. City Nursing & Maternity Home Research Center, 21, Minhajpur, Allahabad- 211003, India., 3. Professor and Head, Department of Biochemistry & Immunology, Subharti Medical College, S. V. S. University, Meerut250005, India, 4. Associate Professor, Department of Pathology, Subharti Medical College, S. V. S. University, Meerut- 250005, India Email: [email protected] Objective: We evaluated, if cord blood of pregnant women with oxygen radical disease had different total enzymatic antioxidant status than those without preeclampsia (PE). Methods: Blood from 23 normotensive (control) and 23 preeclamptic mothers was tested for Malondialdehyde (MDA); and antioxidant enzymes, Superoxide dismutase (SOD); Glutathione Peroxidase (GPx) and Catalase activity and results were compared for oxidative and anti-oxidative system in maternal and cord blood of pair-matched mother and neonate. Results: MDA in preeclamptic maternal plasma was significantly high (p < 0.001) as compared to control. Interestingly, its content in preeclamptic cord blood was significantly low (p < 0.001) compared to their pair-matched maternal blood. SOD activity was 8.7% higher in cord as compared to pair-matched normotensive maternal blood which was significant (p=0.01) whereas in Abstract No: 11.16 preeclamptic cord the level decreased significantly (p=0.011) in comparison to pair-matched preeclamptic maternal. GPx was 16.4% higher in normotensive cord compared to maternal blood and 7% low in preeclamptic cord compared to pairmatched maternal blood. The increase was significant (P=0.011) in normotensive cord whereas in preeclamptic cord the decrease was insignificant (p=0.06). Catalase activity showed 20.97% elevation in normotensive and 16.12% increase in the preeclamptic cord blood compared to their pair-matched maternal blood. This was significant with p=0.01 and p=0.017 in control and preeclamptic group respectively. Conclusion: we found that the oxidative stress is low in the blood of neonates born to preeclamptic mothers. Further studies are needed to explore strategies so that the normal levels of antioxidant are maintained to combat preeclampsia in high risk patients.

Superoxide Dismutase Activity in Ridley-Jopling Leprosy Types

CVB Prasad and MV Kodliwadmath Department of Biochemistry, J. N. Medical College, Belgaum- 590 010, Karnataka, India. E mail: [email protected] Objective: Oxidative stress has been reported in (SOD) activity in Ridley-Jopling leprosy types. leprosy due to poor immunity and malnutrition. Methods: Hundred patients of leprosy comprising The objective of the present study was to 22 Tuberculoid leprosy (TT), 28 Borderline investigate the erythrocyte superoxide dismutase tuberculoid leprosy (BT), 13 Borderline leprosy

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(BB), 16 Borderline lepromatous leprosy (BL) and 21 Lepromatous leprosy (LL) were studied along with 50 normal healthy controls.

enzyme activity was detected along the leprosy spectrum from TT to LL.

Conclusion: The findings suggest diminution of Results: The enzyme activity was found to be SOD activity leading to impaired antioxidant significantly low in leprosy (total patients) as potential in leprosy patients compared to controls. A progressive decline in Abstract No: 11.17

Effect of Wheat Grass on Oxidative Stress in High Fat Diet induced Hyperlipidemia in Rabbits

Dr. Kiran Dahiya*, Dr. Veena Singh*, Dr. Sangeeta B.Singh*, Dr. Jyoti Sethi#, Dr. Mridul Yadav# and Prashanta Saha Roy*. *Department of Biochemistry, # Department of Physiology, Pt. B.D. Sharma PGIMS, Rohtak, Haryana, India. Email ID: [email protected] Results: High fat diet was found to produce hyperlipidemia and an increase in oxidative stress as indicated by a significant rise in malondialdehyde levels while antioxidants like glutathione and vitamin C were found to be reduced significantly. Wheatgrass supplementation along with high fat diet produced an improvement in lipids (total cholesterol decreased and HDL-C increased) while significantly reducing MDA levels. A significant increase was observed in reduced glutathione and vitamin C Methods: Thirty rabbits were divided in three levels. groups of ten rabbits each, group I received control diet, group II high fat diet & group III both high Conclusion: Thus wheat grass can act as an fat diet and wheat grass for a period of ten weeks. antioxidant booster and may have a beneficial role The fasting serum samples were analyzed for total in ameliorating hyperlipidemia and associated cholesterol, HDL-cholesterol, malondialdehyde, oxidative stress. reduced glutathione and vitamin C and compared statistically between the groups. Objective: Hyperlipidemia is a commonly encountered health problem. A number of medicinal plants are found effective in treating hyperlipidemia besides the drug therapy. Wheat grass (Triticum aestivium)is used as a general health tonic and has been reported to be effective in some medical disorders though much literature is not yet available. So this study was planned to observe the effect of wheat grass on high fat diet induced hyperlipidemia in rabbits. bstract No: 11.18

Study of Lipid Peroxidation and Anti Oxidant Status in Pre-Eclampsia

Dr.Rajni S* , Dr. S.S. Halyal** and Dr.Jayaprakash Murthy D.S ** *Assistant Professor; **Professor; Department of Biochemistry, J.J.M.Medical College, Davangere , Karnataka [email protected] Background: Pre-eclampsia is a multiorgan disorder characterized by hypertension and

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proteinuria and a complication of pregnancy affecting 2-4% of all pregnancies. Multiple factors,

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including increased oxidative stress and decreased method. The data was then statistically analyzed antioxidant capacity have been associated with using students't test (unpaired). pre-eclampsia. Results: The results revealed that pre-eclamptic Objective: The purpose of this study is to evaluate patients had significant increase in serum oxidative stress in pre-eclampsia by estimating malondialdehyde and decrease in serum vitamin E,superoxide dismutase and erythrocyte reduced serum pro-oxidants and antioxidants. glutathione levels when compared to healthy Methods: The present study included one hundred controls, and these results were more pronounced and ten women, among them forty were healthy in severe pre-eclampsia when compared to mild controls and seventy were clinically diagnosed pre-eclamptic patients. pre-eclamptic cases who were divided into mild and severe pre-eclamptic cases.The samples were Conclusion: The study concludes that the severity estimated for serum Malondialdehyde by of pre-eclampsia is closely related to the degree of thiobarbituric acid method, serum Vitamin E by oxidative stress. This study suggests that a regular Baker and frank method, serum Superoxide evaluation of oxidant and antioxidant status could dismutase by marklund and marklund method and provide an early predictive index of risk factors erythrocyte Reduced glutathione by Beutler et al and further intervention to prevent pre-eclampsia. Abstract No: 11.19

Attenuation of Inflammation and Oxidative Stress by Plumbago capensis in Freund's Complete Adjuvant-Induced Arthritis in Rats

A.I.Charles Dorni2 , Hannah R. Vasanthi1, K.S Jayachandran2.. and G.V.Rajamanickam2 1 Dept of Biochemistry, Sri Ramachandra Medical College, Sri Ramachandra University, Porur, Chennai, India, 2Centre for Advanced Research in Indian System of Medicine(CARISM), SASTRA University, Thanjavur, India Objective: Oxygen derived free radicals and their products are known to play an important role in the pathogenesis of chronic inflammatory disorders such as rheumatoid arthritis. The purpose of this study was to investigate the antiinflammatory and anti-oxidant effects of alcoholic extract of Plumbago capensis (PC) in adjuvant induced arthritis in rats. glutathione peroxidase and an increase in the lipid peroxidation as indicated by the higher levels of thiobarbituric acid reactive substances (TBARSs). Daily treatment of rats with PC (250 mg/kg) and standard anti-inflammatory drug indomethacin (10mg/kg) for 45 days produced a significant attenuation in the inflammatory response and ameliorated the arthritic changes in the joint. The protection afforded by Plumbago capensis (PC) was more pronounced than that of indomethacin and was associated with normalization of the levels of inflammatory mediators and biochemical parameters of oxidative stress.

Methods: Intra-articular injection of FCA produced inflammation of the joint with a peak effect occurring on day 4 where a maximum increase in the levels of paw edema and inflammatory mediators like PGE2, IL2 and IL6 was observed. Conclusion: The pharmacological potential exhibited by the plant drug is probably mediated Results: This was associated with oxidative stress through the phyto-constituents such as flavonoids with a marked reduction in the levels of in the extract which was also confirmed through glutathione, catalase, superoxide dismutase and in-silico analysis.

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Abstract No: 11.20

Effect of Vitamin E Supplement on Blood Anti-oxidant Status in Cigarette Smokers.

Juhi Aggarwal and Sadhana Sharma Santosh Medical College and Hospital, Santosh University, Ghaziabad-201009, India Background: Coronary Heart Disease (CHD) is a major cause of mortality worldwide and smokers are at high risk of developing CHD. Reactive free radicals are associated with the formation of atheromatous plaques. Cigarette smoke contains free radicals which initiate and propagate the process of lipid peroxidation.Present study aims to assess indices of free radical mediated peroxidation in smokers and non smokers along with the effect of vitamin E supplementation. Methods: Two experiments were designed for the study comprising age group of 30-45 years. Exp.1 has 10 male smokers and 10 non smokers. Plasma and erythrocyte antioxidants and antioxidant enzyme status were measured. Exp. 2 included 20 male smokers and 20 non smokers and each given a placebo followed by vitamin E (1000 mg tocopherol acetate) for 14 days. Plasma indices of lipid peroxidation were measured. Abstract No: 11.21 Results: For Exp. 1 plasma concentration of vitamin E were decreased (p < 0.04) in smokers. Plasma ascorbic acid concentration was 50% lower in smokers but dehydroascorbic acid concentration was raised (p < 0.06) in them. Erythrocyte glucose -6 phosphate dehydrogenase and glutathione peroxidase levels were significantly lower (p < 0.05 and p < 0.02 respectively) in smokers. For Exp.2 In smokers, the elevated erythrocyte lipid peroxidation (p< 0.01) was abolished by vitamin E supplementation (p < 0.05) and conjugated diene levels were raised significantly (p < 0.04). Conclusion: Hence it is evident that free radicals involved in cigarette smoke contribute to the pathogenesis of CHD and increased antioxidant intake inhibits the formation of atheroma.

Oxidative Stress and Antioxidant Therapy in Nephrotic Syndrome

1

Jyoti Dwivedi 1 and Dr.Purnima Dey Sarkar 2 Deptt. of Biochemistry, S.S. Medical College Rewa (M.P.) 486001 India, 2 Deptt. of Biochemistry, N.S.C.B. Medical College Jabalpur (M.P.) India. Objective: The free radicals have a negative influence on renal tissue in nephrotic syndrome. Nephrotic syndrome is a consequence of an imbalance oxidant/antioxidant status. Hyperhomocyst(e)inemia induce oxidative modification in nephrotic syndrome. Zinc and copper deficiency in nephrotic syndrome related to increase urinary zinc and copper losses. The aim of the present study was to estimate the serum total antioxidant capacity, malondialdehyde, homocysteine, copper, zinc, plasma vitamin C during remission and complication.

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Methods: The present study was conducted on 3 groups, group1 comprised of 50 pretreated nephrotic syndrome patients, group 2 comprised of 50 post treated nephrotic syndrome patients and group 3 comprised of 41 secondary nephrotic syndrome patients. Serum total antioxidant capacity, malondialdehyde, homocysteine, copper, zinc, plasma vitamin C were analyzed. Results: In the present study observed there were significant decreased level of serum total antioxidant capacity, copper, zinc, plasma vitamin

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C and increased serum level of malondialdehyde, homocysteine, in group 3 when compared group1. However significant reduction in malondialdehyde, homocysteine and significant improvement in serum total antioxidant capacity, copper, zinc & plasma vitamin C were observed after antioxidant, minerals & B-complex therapy with routine prescription in group 2 when compared to group 1. Abstract No: 11.22

Conclusion: In the present study oxidative stress is higher in secondary nephrotic syndrome than nephrotic syndrome. The study further reports beneficial effects of antioxidants, minerals and Bcomplex vitamins on oxidative stress in nephrotic syndrome patients may prolong need for treatment of nephrotic syndrome and secondary nephrotic syndrome patients.

Antioxidant Status in Patients with Uncomplicated Type 2 Diabetes Mellitus

K. Sowmya1, S.Thanikachalam2 and Satyajeet Giri2 1-Department of Biochemistry, 2- Department of cardiology, Sri Ramachandra Medical College and Research Institute, Chennai- 6000116,India. Email:[email protected] Background: In the past few decades, type 2 diabetes mellitus (T2DM) has rapidly increased worldwide. Persistent hyperglycemia leads to increased generation of reactive oxygen species, as a result the antioxidant capacity is expected to be decreased in diabetic patients. hemolysate for SOD were used. Plasma glucose, lipid profile and uric acid were analyzed in Dimension RxL autoanalyzer using kits supplied by Siemens Dimension. HbA1c was analyzed by Biorad D-10. SOD was analyzed in Hitachi U-2001 spectrophotometer .TAS was analyzed in Konelab 60i autoanalyzer using kits supplied by Randox . Objective: Hence this study was taken up to Statistical analysis was done using students t- test. compare the levels of plasma total antioxidant P < 0.05 was considered significant. status (TAS), uric acid and superoxide dismutase (SOD) of T2DM patients with healthy individuals Results: Significant dyslipidemia & elevated along with routine plasma fasting & postprandial plasma glucose levels were seen among the T2DM glucose, HbA1c and lipid profile. cases. The levels of TAS & SOD were significantly decreased in cases compared to the normal group Methods: The study group comprised of 50 and uric acid levels were found to be slightly healthy individuals as controls and age matched elevated among cases. 35 uncomplicated Type2 diabetes mellitus patients as cases. Samples were collected using BD Conclusion: Our study finding shows reduced Vacutainer tubes. Fluoride plasma for glucose, antioxidant activity among patients with Type2 heparinized plasma for TAS, EDTA whole blood diabetes mellitus. Hence they are prone for for HbA1c, serum for uric acid & lipid profile and oxidative damage due to increased oxidative stress. Abstract No: 11.23

Chronomics of Oxidants, Anti-oxidant Enzymes and Related Molecules in Gyenecological Malignancies

R K Singh, S Singh, R Singh, S Pandey, S Mehrotra, U Singh, O Schwartzkopff, G Cornelissen and F Halberg Departments of Biochemistry and Obstetrics & Gynecology, C S M Medical University and Halberg Chronobiology Center, University Of Minnesota, Minneapolis, MN, USA.

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Background: Chronomics ( an outgrowth of chronobiology, the study of diversity in time ), is the inferential statistical mapping, "imaging'' of time structures in variables in and around us, consisting of rhythms, chaos and trends i.e., of the chronome. The chronome ( from chronos, time, and nomos, rule; time structure ) of oxidants and anti-oxidant defense mechanisms may relate to prevention and curative chronochemotherapeutic efficacy and management. Methods: Forty newly diagnosed women with gyenecological malignancies, 30-60 years of age, and 30 age-matched clinically healthy women were synchronized for 1 week with diurnal activity from about 06:00 to about 22:00 and nocturnal rest. Breakfast was around 08:30, lunch around 13:30 and dinner around 20:30. Drugs known to affect the free-radical system were not taken. Blood samples were collected at 6-h intervals for 24 hour under standardized conditions. Plasma malondialdehyde (MDA), total lipid Abstract No: 11.24

hydroperoxide (LOOH), protein carbonyl (PC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) activities, and serum ascorbate, urate and high-density lipoprotein cholesterol (HDL-C) concentrations and urinary MDA and Melatonin were also determined. Results: A marked circadian variation was demonstrated for each variable in each group by population-mean cosinor (p < 0.01). In addition to anticipated differences in overall mean value (MESOR), patients differed from healthy volunteers also in terms of their circadian pattern. Conclusion: Mapping the broader time structure (Chronome) with age and multifrequency rhythm characteristics of anti-oxidants and pro-oxidants is needed for exploring their putative chemotherapeutic role as markers in cancer chronoprevention and management of gyenecological malignancies.

Seminal Malondialdehyde, Nitric Oxide, Zinc and Superoxide Dismutase Activity in Male Infertility

Kavita More*, Prof P G Samant** and Dr Z G Badade*** *Lecturer, **Professor, ***Professor& Head, Department of Biochemistry, MGM Medical College, Kamothe, Navi-Mumbai, Pin-410 209: [email protected] Objective: The aim of our study was to determine the levels of malondialdehyde (MDA), nitric oxide, zinc and superoxide dismutase (SOD) activity in seminal plasma and to find out their correlation with semen parameters. Methods: The material represents 60 semen samples, which were divided into two groups: Group I-Normospermic (n=30), Group IIOligospermic (n=30). Seminal plasma malondialdehyde, nitric oxide, zinc and SOD were estimated by spectrophotometric methods. The study includes determination of selected semen parameters.

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Results: Seminal plasma malondialdehyde and nitric oxide were high in oligospermic than normospermic. Seminal plasma zinc and SOD were low in oligospermic than normospermic. There was a negative correlation between malondialdehyde and nitric oxide with zinc and SOD in oligospermic. Seminal plasma malondialdehyde and nitric oxide showed a negative correlation with sperm count and sperm motility. Seminal plasma zinc and SOD showed a positive correlation with sperm count and sperm motility in oligospermic group.

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Conclusion: The increase in malondialdehyde, nitric oxide and decrease in zinc and superoxide dismutase levels in oligospermic may cause disruption in the membrane integrity of spermatozoa and may have role in reduction of sperm DNA integrity. There was a positive correlation between zinc and SOD with sperm count and sperm motility, indicating zinc may play Abstract No: 11.25

an important role in spermatogenesis and steriodogenesis. Thus estimation of seminal malondialdehyde, nitric oxide, zinc, and SOD can be useful tools for determining sperm fertilization potential. These parameters could assist in diagnosis of male infertility and help in treatment of infertile male.

Study of Nitrate + Nitrite and Antioxidant Enzyme Levels in Oral Cancer

Kinjal R. Patel, Jayendra B. Patel, Shruti R. Patel, Shilin N. Shukla and Prabhudas S. Patel Biochemistry Research Division, The Gujarat Cancer & Research Institute, Asarwa, Ahmedabad 380 016, Gujarat E mail: [email protected] activities of erythrocyte SOD and catalase were higher in WHT than NHT. Erythrocyte SOD and catalase levels were higher in WHT and patients with OPC as compared to NHT. The erythrocyte SOD and catalase activities were lower in oral cancer patients than patients with OPC. The erythrocyte SOD activity was higher in advanced oral cancer than the early disease. Erythrocyte catalase activity was lower in poorly differentiated tumours than well and moderately differentiated Methods: Blood samples were collected from tumours. Person's correlation analysis revealed that the subjects. NO 2 +NO 3 (nitrite+nitrate) , alterations in plasma NO 2+NO 3 levels were superoxide dismutase (SOD) and catalase levels negatively associated with changes in erythrocyte were estimated by highly specific SOD activities. spectrophotometeric methods. Statistical analysis was done by SPSS statistical software version 11. Conclusion: The data revealed that the alterations in antioxidant activities were associated with Results: Mean plasma NO2+NO3 levels were production of nitric oxide in oral cancer, which elevated in patients with OPC and oral cancer may have significant role in oral carcinogenesis. patients as compared to the controls. Mean Background: Oral cancer is the leading malignancy in India. Nitric oxide and antioxidant enzymes play an important role in etiology of oral cancer. Therefore, the present study evaluated nitric oxide and antioxidant enzyme levels in healthy individual without tobacco habits (NHT, N=30) and healthy individuals with tobacco habits (WHT, n=90), patients with oral precancers (OPC, n=15) and oral cancer patients (n=126). Abstract No: 11.26

Oxidatant and Antioxidant Defence Mechanism in Chronic Myeloid Leukemia

Rizwan Ahmad, Anil K Tripathi, Payal Tripathi, Vinod Kumar Singh and Raj K Singh Departments of 1Biochemistry and 2 Hemato-Oncology Unit, Department of Medicine, C.S.M. Medical University, Lucknow, India Background: Chronic myeloid leukemia (CML) is a myeloproliferative disorder with a

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characteristic genetic rearrangement, the Philadelphia chromosome. Oxidative stress, a pervasive condition of increased amount of free radicals is now recognized to be prominent feature of various diseases including leukemias and their progression. The relationship between the levels of well known oxidative stress markers and antioxidants status reflect better health indices and postures. The present study was planned to review the role of oxidative stress and antioxidant defense system in patho-biology of CML. Oxidative stress was assessed in terms of malondialdehyde (MDA), total lipid hydroperoxide (LOOH) and protein carbonyl (PC) content whereas antioxidant status was evaluated in term of reduced glutathione (GSH) and total thiol (T-SH) and total antioxidant status (TAS) levels in plasma of CML patients. Melatonin (MEL) level was measured in urine. Methods: The present study included 82 (male: female; 1.8:1) CML patients and 70 (male: female; 1.5:1) age-sex matched healthy volunteers. Out of 82 CML patients, 66 were in chronic phase (CML-CP) and 16 in accelerated phase (CMLAP). The median age of CML patients was 35 years and that of healthy participants 34 years. Oxidative stress and antioxidant defense system markers in plasma were evaluated by spectrophotometric procedures whereas MEL level was determined in terms of 6sulphatoxymelatonine excreted in urine by ELISA kit (IBL-Hamburg). Results: There was a significant increase (p<0.05) in plasma MDA, LOOH and PC levels in CML patients as compared to healthy subjects. Our results also showed that plasma MDA, LOOH and PC levels were markedly elevated (p<0.05) in both

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CML-CP and CML-AP as compared to healthy volunteers. Antioxidant defense system which was measured in term of reduced GSH, T-SH, TAS and MEL was found to be significantly decreased (p<0.05) in CML patients and its phases (CMLCP, CML-AP) as compared to healthy participants. During the follow-up of total 66 CML-CP patients for 12 months, 15 patients of CML-CP progressed to the accelerated phase whereas 51 patients remain in CML-CP phase. The mean plasma levels of MDA, LOOH and PC in patients with CMLCP who progressed to CML-AP were found to be higher than in patients with CML-CP who did not progress to the accelerated phase. An elevation in the plasma levels of MDA, LOOH, and PC was observed in CML-CP patients who progressed to CML-AP. The antioxidant defense system in patients with CML-CP who advanced to CML-AP was found to be decreased than in patients with CML-CP who did not progress to the accelerated phase. The antioxidant defense profiles remained decreased in those CML-CP patients who progressed to CML-AP. Conclusion: It could be implicated that plasma MDA, LOOH and PC levels may reveal the magnitude of oxidative stress in CML patients whereas reduced GSH, T-SH, TAS and MEL explain the antioxidant defense system against oxidative stress. All these parameters for oxidative stress and antioxidant defense mechanism may precisely reflect the proliferative signal transduction, disease phenotype and its subsequent disease progression. Plasma MDA, LOOH and PC may serve as indices for oxidative stress and disease progression in patients with chronic myeloid leukemia whereas antioxidant defense system plays an important role in nullifying the oxidative stress.

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Abstract No: 12.1

Antioxidant Markers and Liver Function Parameters in Chronic Alcoholics: A Comparative Study between Koraga Tribal Community, Civilized Alcoholics and Healthy Controls

Naureen Anwar1, Prasiddha Tilak1, Mahesh S1 and Dr Mungli Prakash1 1 Department of Biochemistry, Kasturba Medical College, Manipal, Karnataka-576104; E-mail for correspondence: [email protected] Objective: It is well established that long-term alcohol consumption leads to liver cirrhosis. Sufficient work has been done on biochemical markers of liver damage and antioxidant status of civilized chronic alcoholics. In the current study chronic alcoholics from a tribal community called Koraga are analysed for the same parameters in a view to assess the extent of liver damage as compared to healthy controls and civilized alcoholics. Methods: Serum and urine samples from Koraga alcoholics (n=28), civilized alcoholics (n=30) and healthy controls (n=31) are analysed for liver function parameters and antioxidant markers. Liver function parameters are determined by automated analyzer. Markers of antioxidant status are estimated spectrophotometrically. The data are analysed using SPSS (14.0) urine GST in both civilized and Koraga alcoholics (p<0.01). Serum thiol levels decreased in Koraga subjects significantly (p<0.01) whereas the decrease was not significant in civilized subjects. There was significant increase in urine thiols of civilized alcoholics (p<0.05) compared to controls, however, such finding was not observed in Koraga subjects. On Pearson correlation, serum GST correlated positively with AST, ALT, total bilirubin and urine GST (p<0.01), serum thiol levels correlated negatively with AST and ALT (p<0.05).

Conclusion: Results of our study possibly indicate that the extent of alcohol induced liver damage in Koraga subjects is comparatively lower than civilized alcoholics, even though the alcohol consumption is found to be higher in them. There may be some mechanism that is rendering them resistant to alcoholic liver damage which needs to be explored through further studies at molecular Results: Compared to healthy controls there was level. significant increase in AST, ALT, serum GST and Abstract No: 12.2

Oxidative Stress in Organophosphorous Toxicity

*Dr.M.Jaiprakash Babu Professor & Head of Biochemistry, Rajiv Gandhi Institute of Medical Sciences, Srikakulam, A.P Objective: India is agriculture based country, there is more use of organophosphates leading to various abnormalities, which are largely caused due to the oxidative stress. The aim of the study is to assess oxidative stress and antioxidant status in organophosphorous toxicity in both acute and chronic exposure cases. Methods: The study group consists of 40 farmers who have been involved mainly in spaying activities in the paddy fields for more than five years from near by villages as chronic exposure cases and 20 cases of oganophosphorous poisoning cases as acute exposure cases. The control group consists of

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30 volunteers from the same place with same sex and age and did not exposed to organophosphates.. Blood samples were collected and analyzed for Malondialdehyde (MDA), Superoxide dismutase (SOD) and vitamin C. The observed values were compared with control group. Statistical analysis was performed by analyzing the data using student "t" test and ANOVA. Results: In the present study MDA levels were significantly elevated in Acute Cases (p < 0.001) Abstract No: 12.3

and in chronic cases (p <0.001) due to the inhibition of Acetyl choline esterase and production of oxidative free radicals. Superoxide dismutase (SOD) is the first line of defense against reactive oxygen species. SOD levels were increased in acute cases (p < 0.001) and also in chronic cases (p <0.001) due to elevated antioxidant status. Conclusion: Vitamin C significantly decreased in chronic cases than acute cases indicated the low antioxidant status.

Effect of Lemon Grass Oil Mouthwash on Patients with Gingivitis

Meghana Bhandarkar*, Mahima .B.S*, Madhur Agrawal*, Sonal Sukreet* , Dr. Shobha.U.Kamath* and Dr.Meena AnandÏ% * - Department of Biochemistry, Kasturba medical college, Manipal, Ï% - Department of Periodontics, MCODS, Manipal. Objective: The study was undertaken to determine the influence of lemon-grass oil mouthwash on patients with oxidative stress due to gingivitis, by estimation of antioxidant levels in GCF (Gingival Crevicular Fluid) and saliva. Methods: In this study, 40 individuals were divided into 2 groups : A.Clinically healthy people as controls (10) B.Patients with gingivitis (30) The level of total thiols was determined by DTNB and level of superoxide dismutase (SOD) by Abstract No: 12.4 Winter bourn et.al, both before and after the use of the mouthwash. Results: The level of SOD and total thiols were found to have increased significantly in patients with gingivitis in comparison with healthy controls. Conclusion: The lemon-grass oil in the mouthwash is found to be effective in decreasing the oxidative stress. The values of thiols and SOD indicate this and support the conclusion.

Oxidative Stress and Vitamin Antioxidant Status in Rheumatoid Arthritis.

Dr P.B Desai, Dr. Manjunath.S, Dr. Shivraj gowda, Dr. Sumangala Kadi, Dr. Shivprasad and Dr. Chetana.K. Department of Biochemistry, J N Medical college, Belgaum, Karnataka. Email: [email protected] Background: Rheumatoid arthritis(RA) is a chronic multi system disease of unknown cause.

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A number of factors are implicated in the etiology but none are decisively conclusive as causative

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agents. However a major feature seems to be method modified by Karmarkar and Rajgopal, Vit synovial inflammation hypothesized to be E by Quafie et al modified by Baker and Frank mediated by self perpetuated reactive oxygen and VitC by Evelyn and Malloy method. species(ROS). Results :It was observed that MDA levels were Objective: The present study was done ,to significantly elevated(p<o.oo1) along with evaluate the oxidative stress by estimating diminished levels of antioxidant vitamins ie VitA, malondialdehyde (MDA)levels and vitamin VitE and VitC in all patients of RA as compared to antioxidant status by estimating Vitamin A(Vit A), healthy individuals. Vitamin E(VitE) and VitaminC(VitC) in blood. Conclusion: It can be concluded that free radicals Methods: The present study included 40 clinically may be responsible for causing inflammation and diagnosed rheumatoid arthritis patients and 40 age damage to joints. Depletion of vitamins give a and sex matched healthy individuals .10 ml of clue that their substitution in food may have some blood was drawn and MDA was estimated by therapeutic use in RA. Thiobarbituric acid method, Vit A by Basses et al Abstract No: 12.5

Changes in Advanced Oxidation Protein Product (AOPP) in Moderate and Severly Uremic Rats

Merin Iype C*, Subramanya Upadhya**, Sharmila Upadhya** and Gopalakrishna Bhat*** *Department of Physiology, SNIMS, Chalakka, **Department of Physiology and Biochemistry, SGU, Grenada, West Indies, ***Department of Biochemistry, KMC, Manipal. Email: [email protected] Background: Oxidation of proteins leads to the formation of carbonyl compounds. Carbonyl formation may be an early marker for protein oxidation. Products formed by oxidation of proteins are called advanced oxidation protein products (AOPP), which increase in patients with renal failure. Normal group. After the development of uremia, blood samples were collected at 4, 8, 12 & 16 weeks for estimation of protein carbonyls by DNPH method.

Results: There was a significant elevation in Protein carbonyls in both the uremic groups when compared to sham and normal controls at all weeks. Objective: To estimate the plasma levels of In addition the level of protein carbonyl showed a protein carbonyl, an uremic toxins at various significant difference between moderate and severe weeks of chronic renal failure in rats and establish uremic groups in 4, 12 and 16 week groups. their relationship with the severity and metabolism Conclusion: Protein carbonyl is a uremic toxin during renal failure. which increases during renal failure. There is a Methods: Two models of stable uremia was developed A. Moderate uremia B. Severe uremia. progressive increase in the level of protein carbonyl Both these models were compared with two depending on the severity of the condition which groups of controls A. Sham operated group B. is seen for the first time in nephrectimized rats.

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Abstract No: 12.6

Oxidative Stress and Calcium Levels in Senile and Diabetic Catract Patients

Deepa K, * Sumana Kamath, Vivian D'Souza and Nandini M Dept of Biochemistry,* Dept of Ophthalmology Kasturba Medical College, Mangalore , 575001; Email: [email protected] Background: One of the often seen consequences of aging process is the development of senile cataract. Disturbance in balance between the oxidant and antioxidant levels is thought to be one of the risk factor for the process of cataractogenesis. The transparency of lens which depends on the protein structure and optimum concentration of intracellular calcium is altered by the process of oxidative stress. Results: Significant increase in the serum MDA was observed in patients with diabetic cataract. Antioxidant activity was significantly reduced in the serum as well as the lens in these subjects. However there was no significant change in the serum and lens calcium levels. Comparison between mature and immature cataractous lenses showed a significant increase in the calcium and decrease in antioxidant activity. A positive association of lens calcium with serum MDA was Objective: The main objective of the present study observed. was to compare the degree of oxidative stress and calcium level in senile and diabetic cataract Conclusion: Degree of oxidative stress is more in subject. diabetic subjects indicating that they are prone to oxidative stress at much earlier age. Increase in Methods: Study included 25 senile cataract calcium content of mature lenses suggests its role patients of either sex and 25 diabetic cataract in the process of cataractogenesis. patients. Blood samples were collected during preSupplementation of adequate dose of antioxidants operative period. Serum was analyzed for MDA, may be beneficial in delaying this and avoiding total antioxidant activity and total calcium while lens the complications. was analyzed for antioxidant activity, and calcium. Abstract No: 12.7

Oxidative Stress in Erythrocytes and Plasma of Healthy Controls

Nirjala Laxmi Madhikarmi and Kora Rudraiah Siddalinga Murthy Department of Biochemistry, Central College Campus, Bangalore University, Bangalore- 560 001 India Email- [email protected], [email protected] Objective: Oxidative stress is a biochemical disequilibrium produced by the excessive production of free radicals and reactive oxygen species, which provoke oxidative damage to biomolecules. Malondialdehyde is the major and one of the most frequently used indicators of lipid peroxidation product that is mutagenic and tumorigenic. The present study was carried out to determine the oxidative stress in erythrocyte and plasma of healthy human subjects.

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Methods: Blood was collected from thirty (30) healthy individuals (15male and 15 female), age ranging from 23 to 53 years. Lipid peroxidation was analysed by the estimation of malondialdehyde concentration both in erythrocyte and plasma. Malon-dialdehyde concentration was determined by estimating thiobarbituric acid reactive substances (TBARS) by the method of Buege and Aust.

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Results: The mean ages of male and female healthy controls were 30.93 ± 7.85 years and 26.70 ± 6.98 years respectively. TBARS in plasma and erythrocyte showed highly significant correlation with respect to body mass index (BMI). In female, lipid peroxidation was higher in plasma but lower in erythrocytes. On the other hand, in male lipid Abstract No: 12.8

peroxidation was higher in erythrocytes than compared to plasma. Conclusion: Our findings suggest that BMI is one of the governing for the increase of oxidative stress. The potential relationship between TBARS in plasma and erythrocytes was observed in control subjects.

Oxidant and Antioxidant status in Metabolic Syndrome

PVLN Srinivasa Rao*, N. Manasa Kumari*, Alok Sachan**, KVN Raju, M.M.Suchitra*, V. Seshadri Reddy* and Aparna R. Bitla* *Department of Biochemistry, **Department of Endocrinology, Sri Venkateswara Institute of Medical Sciences, Tirupati, A.P. .Email: [email protected] Objective: Oxidative stress is involved in the pathophysiology of diabetes and cardio vascular complications of metabolic syndrome. Endothelial dysfunction which is the key feature of metabolic syndrome and its vascular complication is intimately linked to insulin resistance. This relationship is partly due to oxidative stress. The present study was taken up to evaluate the oxidant and antioxidant status in patients with metabolic syndrome as compared with healthy controls. Methods: Twenty five patients with metabolic syndrome diagnosed on the basis of WHO criteria taken as subjects along with twenty five, age and sex matched healthy individuals. Malondialdehyde (MDA), as an index of changes in lipid peroxidation was estimated as Thiobarbituric acid reactive substances (TBARS), Plasma total antioxidant capacity as Ferric Abstract No: 12.9 reducing ability of plasma (FRAP), Superoxide dismutase activity (SOD) by continuous Spectrophotometric rate determination. Results: A significant increase (p<0.001) in MDA levels in study group was observed when compared to the control group, where as FRAP levels were decreased in study group compared with control group (p<0.001). SOD activity was decreased but not statistically significant in study group when compared with control group. Conclusion: The findings of the present study suggest the presence of oxidative stress in patients with metabolic syndrome which further increase the cardiovascular risk in these patients. Hence management should be aimed at treating the atherogenic factors associated with metabolic syndrome like dyslipidemia, hypertension etc., and also correcting the antioxidant status.

Study of Antioxidant Enzymes of Seminal Plasma in Association with Mental Stress Subjects

R.K Padalkar, V.R Pandhare, P.S Kamble and P.D Zende PD VVPF's Medical College, Ahmednagar, Maharashtra, India, Email:[email protected] Objective: Mental stress, which is responsible for and social problems. It is reported that mental stress various disorders including abnormality in sperm causes abnormality in sperm quality. In the present cell functions, is one of the most important medical study we investigated the impact of mental stress

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in MBBS students due to final examination and after examination on free radical activity and three important antioxidant enzymes of seminal plasma, Superoxide Dismutase (SOD), Glutathione Peroxidase (GSH) and Catalase. Methods: For carrying out morphological and biochemical analysis, semen samples were collected twice from 50 healthy male volunteers, who were third semester students of a medical college, just before and after stress period 12.00 ± 0.50 week final examination and 12.00 ± 0.50 week gap after final examination, venous blood samples were also withdrawn and serum separated for assessment of cortisol levels. Psychological stress of participants was assessed on the basis of questionnaire and elevated serum cortisol level. Abstract No: 12.10

Similarly non stress subjects were evidenced by the study of questionnaire and normal serum cortisol level. Results: The results demonstrated spermatozoa concentrations, motility index and percentage of rapid progressive motility decreased under stress. Moreover, serum cortisol and seminal plasma Lipid Peroxide (LPO) levels were found elevated along with decreased seminal plasma GSH and reduced SOD. Conclusion: From present study our results indicated that mental stress negatively affected semen quality and antioxidants level is positively correlated with semen characteristics and the concentration of antioxidant scavengers.

Role of Mucuna Pruriens on Antioxidant Enzymes of Seminal Plasma in Association of Mental Stress Subjects

VR Pandhare, RK Padalkar, PS Kamble and PD Zende PDVVPF's Medical College, Ahmednagar, Maharashtra, India Email: [email protected] Objective: In the present study we investigated with M. prurines seed powder (5g day-1) orally and the role of Mucuna Pruriens on activityof rest 25 individuals were kept free from dose. antioxidant enzymes of seminal plasma in the Results: The results demonstrated spermatozoa mental stress subjects. concentrations, motility index and percentage of Methods: Semen samples were collected thrice rapid progressive motility decreased under stress. from 50 healthy male volunteers, who were third Moreover, serum cortisol and seminal plasma semester students of a medical college, just before Lipid Peroxide (LPO) levels were found elevated and after stress period 12.00 ± 0.50 week final along with decreased seminal plasma GSH and examination and 12.00 ± 0.50 week gap after final reduced SOD. M. Pruriens significantly examination, venous blood samples were also ameliorated psychological stress and seminal withdrawn and serum separated for assessment of plasma LPO level along with improved spermatozoa concentration motility index and cortisol level. SOD and Catalase activities were rapid progressive motility. Treatment also restored measured in the seminal plasma. Psychological the levels of SOD, Catalase and GSH in seminal stress of participants was assessed on the basis of plasma of stress subjects. questionnaire and elevated serum cortisol level and vice versa. Stress observed in the individuals after Conclusion: From our results it is concluded that the period of 12.00 ± 0.50 week gap after final M. Pruriens not only reactivates the antioxidant examination was arranged into two groups, 25 defense system of infertile male but also helps in each. The first 25 individuals were administered the management of stress and improves semen quality.

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Abstract No: 12.11

Relationship between ROS and Antioxidants in Male Infertility

Parineeta Samant*, Prof P G Samant** and Dr Z G Badade*** *Lecturer, **Professor, ***Professor & Head, Department of Biochemistry, MGM Medical College, Kamothe, Navi ­ Mumbai, Pin-410 209; [email protected] or [email protected] Objective: There is growing evidence that damage to spermatozoa by reactive oxygen species (ROS) has a key role in male infertility. The aim of the present study was to assess seminal plasma Malondialdehyde (MDA), Vitamin C concentration and Catalase activity. These oxidant & antioxidants represents the lipid peroxidation and spermicidal antioxidant activity respectively. We aimed to correlate the relationship of seminal parameters, MDA concentration with Catalase activity and Vitamin C levels in control and study group. Methods: Specimens were divided into two groups: Group I- Normospermia (n=30); Group II-Oligoasthenozoospermia (n=30). Seminal MDA concentration was measured by K Satoh's method. Seminal Catalase activity was measured by A K Sinha's assay. Vitamin C levels are measured by 2,4-DNPH method. Abstract No: 12.12 Results: Seminal parameters, MDA, Vitamin C levels and Catalase activitiy in both groups were compared. MDA concentrations in oligoasthenozoospermic group were significantly elevated. Catalase activities in oligoasthenozoospermic groups were significantly decreased. Vitamin C levels were significantly lower in oligoasthenozoospermic patients, comparing to the levels found in normospermic men. Assessment of ROS, Catalase and Vitamin C may assist in diagnosis and prognosis of male infertility. Conclusion: Treatment strategies must be directed toward lowering of ROS levels to keep only a small amount necessary to maintain normal cell function and improve antioxidant status.

Study of Lipid-Peroxidation and Antioxidant Status in Malaria Patients

Pawan Kare*, B.K.Agrawal* and V.K.Sharma** *Dept. of Biochemistry, **Dept. of Medicine. Gandhi Medical College, Bhopal. Email: [email protected] Objective: Malaria is a major health problem in developing countries accounting for 2-3 million deaths per year. Oxidative stress (Lipidperoxidation) plays an important role in development of malarial anemia. The present study was done to assess MDA as a marker of lipid-peroxidation and SOD as an antioxidant in malaria patients. Methods: The present study included 50 untreated malaria cases compared to 25 age and sex matched healthy control cases. In these patients following investigation were carried out:1. MDA level was estimated by Buege.et. al.(1978) method. 2. SOD activity was estimated by Das.et. al.(2000) method. 3. Hb was estimated by cyan methemoglobin method.

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Results: The MDA level was significantly increased while SOD level and Hb were significantly decreased in both groups of malaria patients as compared to control groups. The MDA level was found to be significantly (P<0.001) increased while SOD level and Hb were found to be significantly (P<0.001) decreased in P.Vivax as compared to control group. The MDA level was found to be significantly (P<0.001) increased while SOD level and Hb were found to be significantly(P<0.001) decreased in P.Falciparum as compared to control group. The MDA level was Abstract No: 12.13

found to be significantly (P<0.05) more increased in P.Falciparum as compared to P.Vivax while SOD level was found to be significantly (P<0.001) more decreased in P.Vivax as compared to P.Falciparum. The Hb level was found to be significantly(P<0.01) more decreased in P.Falciparum as compared to P.Vivax. Conclusion: We concluded that lipid-peroxidation is a significant problem in malaria patients. A high level of MDA and decline level of SOD activity and Hb level are associated with malarial anemia

A Study of Oxidative Stress in Cancer Patient Before and After Supplementation of Vitamin C and E

Dr. Pranami Bordoloi ,Dr. Bikash Kr. Chaudhury, Dr. Chandan Kr. Nath, and *Dr. Dipali Das Department of Biochemistry,Gauhati Medical College and Hospital , Guwahati ­ 781032 Assam, Senior Resident Doctor, Department of Biochemistry, NEIGRIHMS, Shillong, Meghalya; Email: drpranamibordoloi @ gmail.com Objective: Cancer is the second leading cause of death behind heart disease. Increased lipid peroxidation is due to the altered ratio between free radicals and antioxidant status. The study was carried out to analyze and evaluate the relationship between antioxidant vitamin C & E supplementation and its effect on the lipid peroxidation in the cancer and normal population. Methods: We estimated Malon-dialdehyde, lipid peroxidation marker colorimetrically by Thiobarbituric acids Reactive Substances (TBARS) and uric acid is by Uricase method. Abstract No: 12.14 Results: Statistical analysis is done by Student`t' test. MDA values in both the study groups decreased significantly after 7 days of vitamin supplementation (p< 0.001). Uric acid values in both the studied group increased significantly after Vitamin Supplementation for 7 days (p< 0.001). Conclusion: The study suggested that MDA value reduced in cancer patients within one week of antioxidant vitamin supplementation but for reduction of uric acid levels longer period of vitamin supplementation is required in both the population.

Biochemical Antioxidative Parameters in Type 2 Diabetic Patients with Cardiovascular Complications.

Prashant Nigam*,B. K. Agrawal*, V. K. Sharma** and Bhawna Bhimte* *Dept. of Biochemistry. ** Dept. of Medicine, Gandhi Medical College, Bhopal. E-mail ­ [email protected] Objective: Prolonged hyperglycemia is the major factor in the pathogenesis of atherosclerosis in

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diabetes, which can lead to cardiovascular complications the generation of reactive oxygen

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species (oxidative stress) may play an important P<0.001),(Vit C P<0.01) decreased as compared role in the aetiology of diabetic complications. to control. In TYPE 2 DM with Hypertension Methods: The present study included 50 Type2 group almost similar observation were found to DM with AMI, 50 Type2 DM with Hypertension TYPE 2 DM with AMI group. Value of MDA was patients with 50 normal control group. In these found to be statistically higher significant groups following investigations were done to (P<0.001) increased while SOD, and TAC was assay antioxidant activity. 1. The activity of statistically significant (P<0.001) decreased. The antioxidant enzyme Superoxide dismutase in value of Vit C was found statically less significant erythrocytes by Das et al method 2000, 2. The (P<0.01) decreased. But statistically insignificant plasma level of Vit C by Omaye et al method 1979, (P>0.05) difference was observed when TYPE 2 3. The lipid peroxidation through measuring DM with AMI group compared to TYPE 2 DM Malondialdehyde level in erythrocytes by Buege with Hypertension group. et al method 1978, 4. The Total Antioxidant Conclusion: It can be concluded that the high MDA and low SOD, Vit.C and TAC levels are capacity by D. Koracevic et al method 2001. Results: In TYPE 2 DM with AMI group value associated with T2DM with AMI and T2DM with of MDA was found to be statistically highly Hypertension hence antioxidants, may be significant (P<0.001) increased while value of administered along with Type2 Diabetic and SOD, Vit C and TAC was significantly (TAC, SOD cardiac therapy. Abstract No: 12.15

Evaluation of Flow Rate, pH, Buffering Capacity, Calcium, Total Protein and Total Antioxidant Levels of Saliva in Caries Free and Caries-active Children ­ an in vivo Study

*Dr. Preethi.B.P, **Dr. Jayaprakash Murthy.D.S and ***Dr. Reshma Dodawad *Assistant Professor; **Professor, Dept of Biochemistry, J.J.M.Medical College, Davangere, Karnataka; ***Asst Prof, Dept of Pedodontics, S.D.M.,Dharwad Karnataka. [email protected] Objective: The purpose of this study was to evaluate the relationship between the physicochemical properties of saliva such as flow rate, pH, buffering capacity, calcium level, total protein and total antioxidant levels in caries free and caries active children. capacity, total protein, calcium and total antioxidant capacity. The data was then statistically analyzed using students `t `test {unpaired}.

Results: The results revealed that when all these parameters were compared among the caries free and caries active children, the flow rate, pH and Methods: The present study included one hundred buffering capacity were slightly reduced in caries and twenty healthy children who were divided into active children, but the total protein and total two groups; Group- I and Group- II comprising antioxidant capacity of saliva increased of age groups 7 to 10 and 11 to 14 years significantly in caries active children and the total respectively. Both the groups were then sub- calcium decreased significantly in caries active divided equally according to gender. They were children. further divided into caries free and caries active children with 15 children in each group. Conclusion: Within the limitation of this study, Unstimulated saliva was collected by suction we can conclude that, the physicochemical method and flow rates were determined. The properties of saliva play a major role in the samples were then analyzed for pH, buffering development of caries

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Abstract No: 12.16

Comparative Study of Free Radical Activity of Plasmodium Falciparum and Plasmodium Vivax Malaria Patients

PS Kamble, RK Padalkar, SR Kharade, PD Zende and VR Pandhare PDVVPF's Medical College, Ahmednagar, Maharashtra, India; Email: [email protected] Objective: The present study was undertaken to study the free radical activity in the malaria patients suffering from Plasmodium Falciparum (n=14) and Plasmodium Vivax (n=21) against the control subjects. Plasmodium Vivax infected patients and healthy controls. Serum Ceruloplasmin / Lipid Peroxidation ratio are found to be significantly lower in the malaria patients (p<0.001) when compared with controls. While correlation between them (r=0.521) was direct positive in Plasmodium Methods: Lipid Peroxidation was assessed by Falciparum malaria as compared to Plasmodium measuring serum lipid peroxide in both groups of Vivax and healthy controls. malaria patients by using Malondialdehyde as standard. The serum Ceruloplasmin is also studied Conclusion: The parallel changes were due to along with serum Lipid Peroxidation. different mechanism. Increase in serum Lipid Peroxidation was due to overactivity of free Results: The level of serum Lipid Peroxidation radical, which corresponds with severely tissue were significantly high (p<0.001) in patients with damage while increase in serum Ceruloplasmin Plasmodium Falciparum malaria as compared to was due to acute phase response in malaria. Abstract No: 12.17

Alterations in Oxidant and Antioxidant Status in Early Alcoholic Liver Disease

Ms. Sripradha. R, Dr. M.G. Sridhar and P.Doureradjou-JIPMER Objective: Oxidative stress plays a crucial role in the pathogenesis of Alcoholic liver disease.(ALD). The present study was undertaken to evaluate, compare and correlate the oxidative stress parameters with the enzymatic changes in the serum of ALD patients and healthy controls. commercial kits adapted to auto analyzer respectively.

Results: ALD patients showed significantly higher values of MDA, PC, TSA, GGT, AST and significantly lower GSH level. There was increased ALT, decreased PSA and the values were Methods: The study included thirty ALD patients not statistically significant. Significant correlation and equal number of matched controls. was observed among MDA, PC, TSA, GSH, GGT Malondialdehyde (MDA) was estimated by and AST. Thiobarbituric acid method, Protein carbonyls (PC) by modified Levine's method, Total and Conclusion: The study revealed that oxidative Protein bound sialic acid(TSA, PSA) levels by stress is strongly associated with changes in both modified Aminoff 's method, reduced sialic acid and enzymes in ALD. Further studies glutathione(GSH) by Dithiobis-2nitrobenzoic acid are required to explore the relevance of these method and enzymes (GGT,AST and ALT) by markers for prognostic purposes.

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Abstract No: 12.18

Respiratory Burst Enzymes of Leukocytes in Withaferin Treated Carcinoma of Uterine Cervix

Reshma K, AV Rao, Dinesh M and Vasudevan D.M Department of Biochemistry, KMC, Mangalore Objective: Withaferin,an active component obtained from the dried root extracts of withania somnifera (ashwagandha), showed antitumor and radiosensitising effects in animals. A similar approach in human cancer patients could probably increase the therapeutic outcome. Respiratory burst enzymes of the leukocytes are involved in a delicate interplay of generating and scavenging free radicals. Radiotherapy (RT), could influence the activity of these enzymes by generating free radicals. Furthermore, radiosensitisers also sensitise the tumor tissues to RT by similar mechanisms. cervix , which would perhaps aid in the understanding of radiosensitising mechanisms.Blood samples were collected from stage IIIB carcinoma of uterine cervix patients (n=20), before starting treatment,after 15 days and after 30 days of treatment with RT and withaferin. Age and sex matched controls (n=25)were also considered for comparison.

Results: A significant decrease in NADPH oxidase was observed in baseline samples of cancer patients when compared to controls.Except for this ,there was no change in the activities of other enzymes, when cancer patients were compared to Methods: Therefore a study of the respiratory controls or when baseline values were compared burst enzymes namely NADPH oxidase with that of follow up in case of patients. Myeloperoxidase (MPO), Glutathione (GSH),Glutathione peroxidase (GSH-PX), Conclusion: Based on the results of this study, it Glucose 6 phosphate dehydrogenase (G6PD) and appears that the mechanism of action of withaferin Superoxide dismutase (SOD) were undertaken in is yet to be explored leukocytes of patients with carcinoma of uterine Abstract No: 12.19

Advanced Oxidative Products of Proteins and Total Antioxidants in Gastric Carcinoma Patients

Anita, Sudha K, Beena V Shetty and Gayatri M Rao Dept. of Biochemistry, KMC, Mangalore Objective: It has been suggested that oxidative stress, defined as a shift in pro-oxidant/oxidant balance towards oxidants has been associated with the pathogenesis of many diseases including carcinogenesis. Reactive oxygen species can induce carcinogenesis via injury to macromolecules such as DNA, carbohydrates and proteins. Methods: 25 primary gastric carcinoma patients and 30 healthy control were included in the study. Advanced oxidation protein products, total protein, albumin and total antioxidant activity in plasma were estimated. Results: Our studies demonstrated a significant increase in AOPP, A:G ratio(p=0.003,) and total antioxidant activity(p=0.014).

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Conclusion: The findings suggest that gastric carcinoma is associated with oxygen derived free radicals accumulation and depletion of total Abstract No: 12.20

antioxidant activity has lead to oxidative stress and advancement of oxidative-antioxidative disorders followed by progression of gastric cancer.

Protein Oxidation and Antioxidants in Oral Cancer

Sudha K, Vinodchandran, Gayathri M Rao and Beena Shetty K.M.C., Mangalore Objective: The complex series of cellular and molecular changes that occur through the development of cancer can be mediated by a diversity of endogenous and environmental stimuli.Free radicals have long been known to be mutagenic. albumin.Data compared with that of age and sex matched controls.

Results: The results indicate that there is a significant increase in protein oxidation and a significant decrease in total thiols in oral cancer patients compared to normal subjects.Plasma GSH Methods: This study examines the role of free and albumin also showed a significant decrease radical induced protein oxidation and antioxidant in these patients compared to healthy controls. defence in oral cancer patients.Plasma advanced Conclusion: We conclude that protein oxidation oxidation protein products (AOPP) was estimated and decreased antioxidants can be considered to as protein oxidation marker.Antioxidant status was be important players in multi mechanistic evaluated by determining GSH,thiols and pathogenesis of oral cancer. Abstract No: 12.21

Oxidative Stress in Maternal and Cord Blood of Pre Term Infants

Revathi.R, Saravanan.A, Ramakrishnan.T, Bharathy.N and Sujitha.S Department of Biochemistry, SRM Medical College and Research Centre ­ Chennai Background: Oxygen is an essential component of living organism but formation of ros (reactive oxygen species) seems to be common place in aerobically metabolizing cells. In healthy human body, ros and anti-oxidans remain in balance but when balance gets disrupted, ros increases and leads to occurance of oxidative stress. This oxidative stress has been implicated in enormous variety of physiological and pathological disorders. Objective: The main objective of the present study is to estimate malondialdehyde (MDApreoxidant), anti oxidant enzymes SOD (superoxide dismutase) & CAT (catalase) in maternal and cord blood of preterm infants compared to that of normal individuals.

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Methods: The study group of 25 mother -infant pairs at term delivery (test groups) was selected for the study. Serum samples were analyzed for MDA, SOD and catalase. Student `t' test was used for statistical analysis. Results: There was a significant increase in lipidperoxidase MDA level in preterm motherinfant pairs compared to term (p>0.001), The reduced levels of SOD and CAT observed in preterm mother -infant pair compared to term mother- infant pair. Conclusion: The result of present study indicates that there is a increased oxidative stress in preterm infant -mother pair compared to term motherinfant pair.

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Abstract No: 12.22

Effect of Renoprotective Therapy on Plasma Thiols and Ferric Reducing Antioxidant Power in Children with Nephrotic Syndrome

Rimshida Malik, Priya Pai, Dr. Krishnananda Prabhu, Dr. Nalini Bhaskaranand, and Dr. Anjali Rao Department of Biochemistry , Kasturba Medical College Manipal, India-576104. E mail: [email protected] Ph: 09986353006 Background: Oxidative damage by free radicals has been implicated in number of clinical disorders including renal injury. Reactive oxygen species promote cell injury by lipid peroxidation which disrupts the structural integrity of the tubular epithelial cells and increases glomerular permeability to proteins along with alteration of glomerular hemodynamics. In the present study, biomarkers of oxidative stress like protein thiols and ferric reducing/ antioxidant power were estimated in children with nephrotic syndrome before onset of renoprotective therapy and after 4 yrs of treatment. and after a period of four years of renoprotectivetherapy (enalpril, aspirin, and dipyridamole) and their respective levels were compared. Those patients who recived oral prednisolone (40-60 mg/m2/day) for period of 4 weeks but did not go into remission phase were included in the study. Results: We observed a significant decrease in both plasma protein thiols and ferric reducing/ antioxidant power (p < 0.001) after treatment as compared to their initial values.

Conclusion: This may indicate that antioxidant Objective: To evaluate the effectiveness of status of these children was decreased as there was renoprotective therapy on the levels of these a decreased demand due to improvement in clinical state parameters. Methods: Plasma protein thiols and ferric reducing/antioxidant power were measured in children suffering from nephrotic syndrome attending Kasturba Medical College Hospital, Manipal before the onset of renoprotective therapy References: 1. Contrib Nephrol. 2008;160:17-28 2. Clin Chim Acta. 2002; 325: 147 -150 3. Am J Med. 1991; 91: 31S ­ 38S 4. Arch Dis Chil. 200; 82: 76 ­ 78

Abstract No.12.23 Role of Lipid Peroxidation and Enzymatic antioxidant Status in complicated Pregnancies: Sadanand B Patil 1, M.V. Kodliwadmath 1 and Sheela M. Kodliwadmath 2 Asst. Professor Professor, Dept. of Biochemistry1 and professor, Dept of Obstetrics and gynaecology2; J.N. Medical College, Nehrunagar, Belgaum-10, Karnataka. Objective: Pre-eclampsia and eclampsia are pregnancy complications with serious consequences for mother and infant. Uncontrolled lipid peroxidation may play an important role in the pathophysiology of pre-eclampsia and eclampsia by causing vascular endothelial cell dysfunction. Antioxidants serve to control lipid peroxidation. We tested that antioxidant protective mechanisms are diminished in women with Preeclampsia and eclampsia. Methods: Lipid peroxidation and antioxidant markers were assayed in 25 healthy non-pregnant women as control, 25 third trimester normal pregnant, 25 pre-eclamptic and 25 eclamptic

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patients of the same trimester by standard spectrophotometer methods. Results: In Pre-eclampsia and eclampsia malondialdehyde, a product of lipid peroxidation was significantly increased while enzymatic antioxidants like superoxide dismutase, Glutathione perxidase Glutathione reductase and catalase were reduced significantly as compared to normal pregnant and non-pregnant controls. Abstract No: 12.24

Conclusion: Lipid peroxidation is an important factor in the pathogenesis of pre-eclampsia and eclampsia. The decrease in antioxidants is probably due to compensatory nature responding to the increased lipid peroxide load in preeclamptic and eclamptic patients and may reflect the severity of the disease. Early attention, intensive management and better treatment with antioxidant vitamins may be essential for improving the maternal and foetal outcome in PIH

Study of Antioxidant Status with Respect to AF after Cardiac Surgery

Shalini Nair*, Khalid Iqbal**, Madhavi S Phadke* and Jagdish Khandeparkar** *Department Of Biochemistry, LTMG Hospital & LTM Medical College, Sion, Mumbai ­ 400022, **Department of Cardiovascular & Thoracic Surgery, LTMG Hospital & LTM Medical College, Sion, Mumbai - 400022 Objective: Atrial Fibrillation (AF) is the most common sustained tachyarrhythmia in patients of valvular heart disease. Post-operative AF continues to be the most common complications after open heart surgery. It is associated with increased cardiovascular morbidity and mortality. Our attempt was to study the role of oxidative stress in patients after cardiopulmonary bypass. collected preoperatively and on the 5th post operative day for the measurement of various antioxidant and oxidative stress markers [glutathione, vitamin C, super oxide dismutase, MDA].

Results: Decrease in the antioxidants and increase in the free radical was evident in all the 3 groups. Glutathione, Super oxide dismutase and Vitamin Methods: Patients undergoing valve procedures C showed a decrease in their levels on the 5th postoperative day as compared to the preoperative were categorized in to 3 study groups namely, value; p<0.001. MDA showed an increase on the 1. Patients who were in normal sinus rhythm 5th day which was more distinct in Gp3 (8.5±3.6 (NSR) pre and post operatively (A1). vs. 11.07±3.46); p<0.001. 2. Patients who converted in to NSR Conclusion: Patients who exhibit AF after cardiac postoperatively from AF (A2). surgery have significantly increased oxidative 3. Patients who were in AF preoperatively and stress, and significantly lower antioxidant status continued to be in AF postoperatively also (A3). which can result in increased myocardial Post operative AF was assessed by 12 lead oxidation. It is also evident that this differential electrocardiogram and required initiation of response is present at the systemic level and not antiarrythmic therapy. Blood samples were just within the myocardium.

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Abstract No: 12.25

Chronic Alcohol Induced Tissue Injury: Microarray Analyses of Gene Regulation in the Disease Process

R. Sandeep, P.Nagababu, Sunitha T, Rekha, D, Farid, M., Malathi, R., .Ranjeetha, N, and R. Polavarapu* Dept. of Biochemistry, Katuri Medical College, Guntur, AP 522019, India, *Emory University School of Medicine, Atlanta, GA 30322, USA Objective: Chronic alcohol abuse is known to have a positive correlation with the onset of tissue injury including liver cirrhosis, pancreatitis, gastitis, acute respiratory distress syndrome (ARDS) and an increased risk for the development of breast cancer. Though the causal relationship between the alcohol abuse and the disease process is not clear but was implicated metabolic activation of ethanol in these tissues might be the causative of the disease process. Here, using alcoholic rat animal model and Affymetrix high density gene chip technology, we report differential gene regulation in different tissues due to chronic alcoholism. Methods: Male and Female Sprague-Dawley rats were pair-fed with isocaloric Lieber-DeCarli liquid diets for six weeks. Different tissues of interest were isolated and subjected for microarray analyses using Affymetrix gene chip technology. Abstract No: 12.26 Results: The data suggests like in liver, there is a state of oxidative stress in those tissues with organ injury and is correlated with phase I and II enzymes that are involved in ethanol metabolism. There was an increase in the expression of alcohol dehydrogenase and several cytochrome p450 gene families in Antrhrum and lung. Several genes involved in glutathione homeostasis increased expression levels, including glutathione Stransferases, several inflammatory proteins and oxidative stress genes are down regulated at the mRNA level. In the pancreas, it was suggested that several oxidative stress genes and genes that are involved in the genesis of inflammation were differentially regulated. Conclusion: This produces an imbalance in the homeostasis of pro and anti-oxidative stress producing, inflammation, reduced immunity and organ damage at large

Involvement of Cellular Oxidants in Deregulated Redox Homeostasis in Diabetes Mellitus

P. P. Singh* and Farzana Mahdi** * Professor Emeritus ** Director Academic's, Department of Biochemistry, Era's Lucknow Medical College & Hospital, Sarfarazganj, Hardoi road, Lucknow. Email [email protected] Background: The debate on reactive oxygen species (ROS) involvement, as a mediator in the genesis of diabetes mellitus (DM) (both in type 1 & 2) and as a critical indicator in development of diabetic complications continues, especially in Indian context which tops the global list of diabetic patients. The moot issues about ROS participation in DM are: a) heightened oxidative stress (OS) is universal or selective. b) cause or consequence. c) central or peripheral. d) facultative or obligatory. e) mechanism/s for activated generation of ROS pathways. f) antioxidants (AO) in the management of the disease. g) why majority of clinical trials have failed to achieve envisaged benefits. The

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studies of our group have revolved around these issues.

glucose induced hyperglycemic spikes are accompanied with raised OS. 5) most importantly Methods: Case controlled studies and clinical experimental induction of chronic or acute OS in trials among several controls and experimental animal models is not accompanied with hyperglycemia distinctively suggesting that OS studies on animal models. alone is not a causative factor. 7) AO treatment Results: Our results do not posit a putative has almost no value in management of patients involvement of OS as measure risk factor in DM with good antioxidant status. As a corollary, several or in associated complication nor benefits of AO pathways can get stimulated to raise ROS treatment in non deficient subjects. production in most of the patients. This may raise Conclusion: We conclude from our data: 1) raised OS to a certain degree, but its contribution to development of DM or further complications is OS or weak AO network is present only in selected only a minor one and only in small number of patients. 2) in majority of patients it begins as cases. AO can not prevent or cure DM with or consequence 3) in humans it does not initiate without complications nor can attenuate it if insulin resistance but may aggravate it. 4) diet or patients have pre existing AO status. Abstract No: 13.1

Oxidative Stress and Inflammatory Response in Pre-eclampsia

Sathish B M 1, Bobby Z 1 and Habeebullah S 2 Department of 1Biochemistry and 2Obstetrics&Gynaecology, JIPMER, Puducherry Objective: To compare markers of oxidative stress Results: MDA, PC, PBS, us-CRP levels were and inflammatory response in pre-eclampsia significantly increased in pre-eclampsia group versus normal pregnant women. compared to the control group. Statistically significant correlation was observed between Methods:Reduced glutathione (GSH), MDA levels and the CRP in the pre-eclampsia malondialdehyde (MDA), protein carbonylation group. PC), protein bound sialic acid (PBS) and ultra sensitive C-reactive protein (us-CRP)were Conclusion: Oxidative stress synergizes with compared between 30 controls and 30 pre- inflammation in pre-eclampsia, which increases the disease severity. eclampsia cases. Abstract No: 13.2

Antioxidant Activity of Fermented Tea (Kombucha)

U. Satyanarayana and T. Srihari. Department of Biochemistry, Siddhartha Medical College, Dr. NTR University of Health Sciences, Vijayawada, Andhra Pradesh. ­ 520008. [email protected] Objective:Kombucha is a home made fermented tea that has been traditionally consumed in China for over 2200 years (Since 220 BC), and is now being used in many other countries. The claimed

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health benefits of kombucha include in disorders such as diabetes mellitus, hypertension, cancer, atherosclerosis, arthritis, constipation, AIDS etc. The biochemical basis of the health benefits of

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kombucha remains largely unknown. The present study was undertaken to find out the total antioxidant activity and total content of phenolic compounds in the kombucha.

Methods: Kombucha was prepared by fermenting freshly made tea after inoculating with a previously grown culture of kombucha. The antioxidant activity of kombucha was determined by ABTS (2,2'-azinobis-3-ethylbenzo-thiazoline6-sulfonic acid) free radical scavenging capacity Conclusion: Further studies are required to of kombucha samples on different days (upto 7 identify the nature of antioxidants in kombucha. days) of fermentation. Total phenolic compounds in kombucha samples were estimated by FolinCiocalteau method. Abstract No: 13.3

Results: Kombucha exhibited good antioxidant and free radical scavenging activity, besides the presence of phenolic compounds. Further, the antioxidant activity and phenolic compounds concentration were found to increase with increased fermentation time. It appears that the claimed health benefits of kombucha consumption may be partly related to the antioxidants produced during fermentation.

Health Benefits of Origanum vulgare (a Medicinal Herb) through its Antioxidant Property.

T. Srihari. U. Satyanarayana and N. Nalini* Department of Biochemistry, Siddhartha Medical College, Dr. NTR UHS, Vijayawada, Andhra Pradesh. ­ 520008, * Department of Biochemistry, Annamalai University, T amil Nadu ­ 608002.Email: [email protected] Objective:The use of medicinal herbs in the healthcare of humans is centuries old. Oregano (Origanum Vulgare L.) is widely used medicinal herb in a treatment of various ailments. The present study was undertaken to evaluate the efficacy of oregano by in vivo studies against 1,2dimethylhydrazine(DMH)-induced rat colon carcinogenesis by estimating circulatory lipid peroxidation (LPO) and antioxidant status. In addition, the in vitro antioxidant activity of oregano was also studied. Methods: DMH-induced cancer was developed in Wistar rats. Oregano was orally supplemented everyday in a dose-dependent manner to different groups of rats for 15 weeks. Tumour-bearing rats showed increased LPO in the circulation which was accompanied by a decrease in levels of antioxidant enzymes (superoxide dismutase, catalase etc.), besides reduced glutathione levels. Oregano supplementation however, reversed the circulatory LPO and antioxidant profile in colon cancer bearing rats. Result:In vitro studies demonstrated that oregano exhibits good free radical scavenging activity, besides the presence of phenolic compounds. Conclusion:The medicinal herb oregano possesses antioxidant property which was proved by both in vitro and in vivo studies. This property of oregano may be involved in the treatment of cancer and other ailments.

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Abstract No: 13.4

Status of Enzymatic and Non-enzymatic Anti-oxidant Systems in Patients with Acute Ischemic Stroke

J. Sudha Rani Assistant Professor, Dept of Medical Biochemistry, GVP Biomedical Engineering College for Women, Visakhapatnam, AP. Objectives: Oxidative stress is probably involved in neuronal damage induced by ischemiareperfusion. The present study was designed to measure the enzymatic and non enzymatic antioxidant status following acute ischemic stroke patients with large vessel and small vessel infarcts. Methods: The study included 72 subjects in which 30 were controls and 42 were acute ischemic stroke cases. They were diagnosed basing on clinical manifestations and by CT scan of the brain. The cases were further divided into 2 groups. The ischemic stroke patients with large vessel infarcts (Group-L) and small vessel infarcts (Group-S) basedCT scan of the brain. Serum enzymatic antioxidants (superoxide dismutase and glutathione peroxidase) and the non enzymatic antioxidants (reduced glutathione, vitamin E and vitamin C) were estimated in all the subjects by using spectrophotometer. Results: Ischemic stroke cases had significantly lower levels of enzymatic and non enzymatic antioxidants, compared to controls which is statistically significant (p<0.05). The Group L ischemic stroke cases had statistically significant (p<0.05) lower levels when compared to Group S ischemic stroke cases. Conclusion: The antioxidant activity of serum may be an important factor providing protection from neurological damage caused by stroke-associated oxidative stress. The present study suggest that reduced antioxidant status may be used as an indirect evidence of oxidative stress induced neuronal damage in acute ischemic stroke which may be useful for monitoring and optimizing antioxidant therapy.

Abstract No: 13.5

Oxidative Protein Damage in Thyroid Dysfunction

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Dr. Sweta Shivshanker1 and Dr. Anushre Prasad2 Tutor, 2Assistant Professor, Department of Biochemistry, Kasturba medical college, Manipal- 576104 Email address: 1 [email protected], 2 [email protected]

Objective: To determine plasma oxidative protein Patients were classified based on T3, T4, TSH damage in patients with thyroid dysfunction. measurements. Methods: 50 hypothyroid patients (mean age 44.6±2.1); 40 hyperthyroid patients (mean age 44.75±2.6) and 50 patients with euthyroid goiter (mean age 37.2±1.6) were included in the study.

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Plasma protein thiols, protein carbonyls, erythrocyte glutathione and glutathione reductase activity were measured spectrophotometrically to determine the oxidative protein damage. Plasma

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protein thiols were measured after oxidation with Ellman's reagent, plasma protein carbonyls measured with DNPH; erythrocyte glutathione measured with DTNB and glutathione reductase activity in haemolysate expressed as activity coefficient.

Results: There was decrease in protein thiol levels in hyperthyroid patients (114.15±2.0 nmoles/ L) and hypothyroid patients (145.8±2.7nmoles/ L) when compared to Euthyroid controls (244.06±3.1 nmoles/ L) p<0.001. Protein carbonyl levels in hyperthyroid patients (1.938±0.03 ng/mg protein) and hypothyroid patients (1.707±0.02 ng/mg protein) were elevated when compared to Conclusion: Both hyperthyroidism and euthyroid controls (1.013±0.01 ng/mg protein) hypothyroidism is associated with oxidative [p<0.001]. Erythrocyte glutathione levels in protein damage in plasma. Abstract No: 13.6

hyperthyroid patients was 11.74±0.09 mg/g Hb which was elevated, when compared to Euthyroid patients (10.147±0.11 mg/g Hb) [p<0.001]. However, erythrocyte glutathione levels were significantly lower in hypothyroid patients (8.72±0.07 mg/g Hb) [p<0.001]. Glutathione reductase expressed as activity co-efficient in hyperthyroid patients was 1.47±0.01 and in hypothyroid patients was 1.23±0.01. The activity co-efficient was increased in both the groups when compared to controls (1.009±0.03) [p<0.001] which indicated a significant reduction of the antioxidant capacity.

Auto Antibodies against Oxidized Low Density Lipoproteins and Lipid Peroxidation in Patients with Essential Hypertension

K.Ramalingam *, K.Santha , S.Sethupathy and R.Vinoth Kumar * * Department of Biochemistry, Narayana Medical College and Hospital, Nellore ,A.P, Division of Biochemistry, Raja Muthiah Medical college and Hospital, Annamalai University, Annamalai Nagar, TamilNadu. Background: Lipid peroxidation has been suggested to play a key role in the oxidative modification of LDL (oxd-LDL) which stimulate the production of auto antibodies by B-cells and anti-oxd LDL antibodies are produced. These antibodies could represent a biological marker of oxidative stress and serve as markers of atherosclerosis. Essential hypertension is a major risk factor for atherosclerosis, recently border line hypertension also has been shown to be a risk factor for atherosclerosis. The aim of this study is to examine the correlation between oxd -LDL antibodies and lipid peroxidation in patient with essential hypertension. healthy individuals (n=60) levels of Malonaldihyde (MDA), Total cholesterol, Triglycerides, HDL and LDL were estimated by spectrophotometry and levels of Oxd- LDL antibodies were obtained by ELISA.

Results: Plasma levels of MDA, anti-oxdLDL antibodies, Total cholesterol and LDL Cholesterol is higher in patients than those in controls. Among patients concentration of MDA, total cholesterol and LDL cholesterol were not significantly different, however the concentration of anti-oxd LDL were higher in essential hypertensive patients (p=0.003). Significant positive correlation was observed between plasma levels of MDA, total Methods: Blood samples were collected from cholesterol, LDL cholesterol and the concentration patients with essential hypertension (n=55) and of anti-oxdLDL in patients but not in the controls.

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Conclusion: High concentrations of anti-oxdLDL suggest an increase in oxidative stress that would contribute to the development of atherosclerosis. The observed correlation of MDA with antiAbstract No: 13.7

oxdLDL may suggest a possible pro-oxidative activity of MDA in patients with essential hypertension.

Oxidative Stress and Insulin Resistance in Type-2 Diabetes Mellitus

R.Viswakumar, K.Ramalingam and R.Vinoth Kumar * Department of Biochemistry, Narayana Medical College and Hospital, Nellore ,A.P Background: Diabetes mellitus (DM) is a metabolic disorder characterized by hyperglycemia with disturbances in carbohydrate, fat and protein metabolism. Hyperglycemia may cause increased production of free radicals, leads to oxidative stress results in cardiovascular and Reno vascular complications. Recent studies demonstrate the reactive oxygen species interfere with insulin signaling cascades which leads to resistance to the insulin action . The present study was aimed to study the relation between insulin resistance and oxidative stress. Methods: Blood samples were collected from patients with type-II DM (n=50) and healthy individuals (n=30) levels of Malonaldihyde (MDA), reduced glutathione,Ferric reducing Abstract No: 13.8 antioxidant power(FRAP), serum creatinine, fasting plasma glucose and postprandial plasma glucose were estimated spectrophotometrically.The levels of fating of insulin were estimated by ELISA. Insulin resistance was calculated by standard formula. Results: Statistical analysis of the results shows MDA and insulin resistance were increased significantly in the patients when compared to control subjects. Reduce glutathione and FRAP were decreased in patients than the controls.. Conclusion: The study conclude that the insulin resistance is associated with oxidative stress. Antioxidants supplements in type-2 diabetes mellitus may reduce the associated complications.

Lipid Profile and Glucose Tolerance Test in Cholelithiasis

R.N Devaki and H.S. Virupaksha Asssistant Professor, Department of Biochemistry, J.S.S Medical College, S.S Nagar, Mysore-570015 Karnataka, E-mail : [email protected] Background: Cholilithiasis is a common disease of gallbladder characterized by gall stone formation.It is associated with abnormalities in the relative concentrations of the major biliary lipids like cholesterol and phospholids. of cholelithiasis with altered glucose tolerance.

Methods: The study was carried out on patients with cholelithiasis. A total 50 patients were taken for the study, among which 35 were female patients and 15 were males. (Age range ­ Objective: 1.To study the association between 30 to 70 years). An equal number of age and serum lipids like total cholesterol, LDL choles- sex matched healthy subjects formed the conterol, HDL cholesterol, triglycerides and trol group. Patient's fasting venous blood was cholelithiasis.2.To study the possible association drawn for the determination of total cholesterol,

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HDL-cholesterol, LDL ­ cholesterol, triglycerides and for fasting blood glucose. A venous blood sample was also drawn after 2 hours of a dose of 75g of oral glucose, to perform glucose tolerance test. Results : There was significant increase in the Abstract No: 13.9

total cholesterol, triglyceride and LDL- cholesterol in patients with cholelithiasis when compared to the control groups. There was significant increase in blood glucose level in female patients but it was not significant in male patients.There was signicant decrease in HDL-cholesterol levels.

Association of Hypoalbuminemia with Size of Pleural Effusion in Pneumonia.

Mrs Ranjita V. Gaur and Dr (Mrs ) Rita M. Shah Email : [email protected], [email protected] Objective: The objective of this study was to assess association of hypoabluminemia & pleural effusion & to propose a possible patho-physiologic mechanism. Methods: The Study group consisted of patients who had presumed bacterial pneumonic complicated by pleural effusion and admitted to Dhiraj General Hospital from March 2004 to April 2006. The Volume of Pleural fluid was assessed semi quantitatively & categorized as small, moderate or large. The following laboratory results were recorded for the study: complete blood count, ESR, Serum electrolyte level, renal & liver function tests, albumin & protein levels. The finding were compared with age & gender, who had matched persons bacterial pneumonic without pleural effusion. Abstract No: 13.10 Results: Of the 75 patients in the study group 23(31%) had small effusion, 24(32%) had moderate effusion and 28(37%) had large effusion. Moderate to severe hypoabluminemia was found in patients 52% of the study group 6% of the patients with pneumonic without pleural effusion. Means serum albumin level was lower in patients with large pleural effusion than in patients with small effusion (2.66+/- 0.37 vs 3.66+/-0.47 g/dL). There was no evidence of albumin loss or significant malnutrition. Estimation of the amount of albumin in the drained pleural fluid suggested an albumin shift from blood to pleural fluid. Conclusion: The Significant hypoabluminemia is common with patient of parapneumonic pleural effusion. Large effusion are associated with low serum albumin levels, which might be explained in part by a shift from blood to the pleural fluid.

Biochemical Effects of Feeding Diglycerol Disulphide in Rats Fed Ethanol or High Fat Diet

Blessy M. I., Vinod P., Praveen K. V., Sheeba V., J. K. Mukkadan and P. K. Joseph Little Flower Institute of Medical Sciences and Research, Angamaly 683 572, India. Email ID: [email protected] Objective: Aliphatic sulphur compounds are implicated as the active ingredients in garlic and onion which have medicinal properties. However they are not acceptable to many because of the harsh taste or smell or because of religious tabor. Hence we have synthesized a synthetic aliphatic disulphide, diglycerol disulphide and tested its biochemical effects. Methods: Thioglycerol was treated with iodine to get diglycerol disulphide. It was extracted with ether. After evaporating the ether, light yellow oily

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liquid obtained had a feeble smell of garlic and ethanol such changes were ameliorated. Similarly taste was not harsh. This was fed to rats along similar changes produced in serum, liver and with ethanol or high fat diet. kidneys by feeding a diet containing 30% dalda could also be prevented by feeding daily 10mg Results: When 3ml 30% ethanol was fed to fasting diglycerol disulphide per 100g body weight. rats, triacyl glycerol, total cholesterol were raised in liver, kidney and serum. Also AST, ALT and Conclusion: Diglycerol disulphide could be useful ALP of the serum were raised. When such rats to produce hypolipidemic effects and to protect were fed 10mg diglycerol disulphide along with the liver similar to garlic. Abstract No: 13.11

The Prevalence of Celiac Disease in Patients Presenting with Gastro-intestinal Symptoms

Satinder Kaur, APS Narang, Navpreet Kaur and Indu Verma Department of Biochemistry Dayanand Medical Collage & Hospital, Ludhiana, Punjab Objective: Celiac disease is inherited, an auto immunologically mediated intestinal intolerance to wheat protein gliadin Celiac disease is associated with chronic gastrointestinal symptoms and malabsorption of nutrients causing weight loss, metabolic bone disease and anemia. the period of ten months were included in the study. Patients were divided into four age groups of 05,5-14,14-40 and > 40 years.

Result:Anti ttg IgA levels were estimated using ORGENTEC diagnostic kits. 16.3% patients were found to be positive. Females (55%) are affected Methods: The early diagnosis of celiac disease is more than males. a set of blood test to measure autoantibodies especially anti tissue trance glutaminase (ttg) IgA Conclusion:Maximum number of patients 3000 male and female patients visiting out patient detected positive are in age group of 14 to 40 years. department (Medicine, Gastrentology, pediatric) Serological screening can be highly useful in of Dayanand Medical Collage and Hospital over detecting sub clinical cases of celiac disease as well. Abstract No: 13.12

Jun N-Terminal Kinase (JNK) mediates Dextran Sodium Sulfate-Induced Tight Junction Barrier Dysfunction in Caco-2 Cell Monolayers.

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Geetha Samak1 and Radhakrishna Rao2, DVS College of Arts and Science, Shimoga, Karnataka, India, 2 Professor, Department of Physiology, University of Tennessee, Memphis, TN. Email ID: [email protected] Objective: JNK /SAPK (Stress activated protein kinase) activation plays an important role in the intestinal inflammation in patients with inflammatory bowel disease (IBD), ulcerative colitis and Crohn's disease. Several types of stress and allergens disrupt intestinal epithelial barrier function and enhance mucosal permeability. The

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major component of mucosal barrier function is epithelial tight junctions (TJs) and therefore, stressinduced inflammatory diseases are likely to be initiated by the disruption of TJs and induction of barrier dysfunction. Methods: Dextran sodium sulfate (DSS)-induced

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murine colitis is widely used model for human colitis/IBD. We investigated the role of JNK in mediating the DSS-induced disruption of barrier function in Caco-2 cell monolayers. Barrier function was evaluated followed with the exposure to 2% DSS by measuring transepithelial electrical resistance (TER) and inulin flux. TJ integrity was assessed by confocal microscopy. Role of JNK in DSS-induced TJ disruption was determined by inhibiting JNK activity by SP600125. Tyrosinephosphorylation of TJ proteins was determined by immunoprecipitation and immunoblot analysis .

redistribution of occludin and ZO-1 from the intercellular junction into the intracellular compartment. DSS rapidly increased the level of phosho-JNK. Pretreatment with SP600125 significantly attenuated DSS-induced decrease in TER, increase in flux and redistribution of TJ proteins. DSS-induced tyrosine-phosphorylation of TJ proteins was inhibited by the pretreatment of SP600125.

Conclusion: These studies indicate that DSS disrupts tight junctions of Caco-2 cell monolayer by JNK activity dependent mechanism which involves tyrosine-phosphorylation of TJ proteins Result: DSS-induced decrease in TER, increase and redistribution TJ proteins from the intercellular in inulin permeability was associated with the junction. Abstract No: 13.13 Storage Stability Characteristics of Hepatobiliary Enzymes in Cattle and Buffalo Divya P.D.* and K.K. Jayavardhanan** MVSc Scholar, ** Associate Professor, Department of Veterinary Biochemistry, College of Veterinary and Animal Sciences, Mannuthy, Thrissur- Pin 680651 Email: [email protected] Results: The study revealed significant differences in the storage stability characteristics of each enzyme in both species at different times and temperatures. The investigation recommended 4 ºC as the ideal storage condition for preserving sera samples of cattle for ALT assay, as it was unstable at freezer, while both 4 ºC and -20 ºC for buffalo serum. Storage of serum at 4 ºC and 20 ºC was suitable for AST assay for cattle whereas buffalo serum showed maximum stability in the serum kept at 4 ºC. The study suggested that ALP assay should be performed on the day of venipuncture itself in both species as the enzyme showed wide variations upon storage. The storage Methods: Blood samples were collected from of serum both at 4 ºC and -20 ºC was ideal for eight adult healthy cattle and buffalo, maintained GGT assay in buffalo whereas 4 ºC was at University farms, Mannuthy. Aliquot of serum recommended for cattle serum. were assayed for ALT, AST, ALP and GGT Conclusion: It is therefore advisable to consider activities after storing at room temperature (25 ºC), stability of serum hepatobiliary enzyme activity 4 ºC and -20 ºC for definite time intervals up to of each species separately for a more valid and two weeks post collection. reliable result. Objective: Although modern analysers can process samples at a rapid rate, when blood samples are collected in large numbers or when many different analysis are required, it is inevitable to store the sera samples and do the analysis on a later date. In veterinary medicine, to date very little has been published on the stability of serum enzyme markers during preservation. The study described here was designed to find out the ideal storage condition for preserving sera samples of cattle and buffalo for the analysis of hepatobiliary enzymes like ALT, AST, ALP and GGT.

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Abstract No: 13.14

IL-6 is a Reliable Independent Biochemical Marker for Predicitng Severity of Acute Pancreatitis

G.Vasantha lakshmi * Pradeep Naik * B. Prem sagar* B. Vijay Kumar** and K.Ravindranath** *Dept of Biochemistry, **Dept of Gastroenterology, GLOBAL HOSPITALS, Lakidikapool, HYDERABAD-500004, Background: There are several biochemical 1261 kit, with a cut-off of 380 pg/ml. The final markers that have been used in an attempt to outcome of the patients in terms of severity of predict the severity of acute pancreatitis. There is pancreatitis viz; mild or severe were noted. lack of clinical data in Indian population. Results: There were 21 patients with severe acute Objective: We performed this study in a tertiary pancreatitis and 32 were mild cases. The mean ILreferral care hospital and aimed at assessing the 6 was 540.57 (SD 761.82). Bivariate analysis reliability of IL-6 as an independent marker in showed IL-6 to be significantly higher in severe predicting severity of acute pancreatitis. pancreatitis. The sensitivity, specificity, PPV, NPV and AUROC of IL-6 in predicting severe acute Methods: A total of 53 patients were included in pancreatitis was 90, 94, 90, 94% and 0.9539 the study. All patients who met the specified respectively. clinical and biochemical criteria of acute pancreatitis and those who presented within 72 Conclusions: IL-6 is a very accurate and reliable hours of the onset of symptoms were included in independent biochemical marker for predicting the study. IL-6 was measured in all these patients severe acute pancreatitis. on admission. We used IL-6-EASIA-CE KAP Abstract No: 13.15

Peritoneal Fluid and Bblood Lactic Acid Levels in Acute Abdominal Disorders

A.P.S. Narang, S Goyal*, Indu Verma, Satinder Kaur and J.S. Multani* Department of Biochemistry and Surgery*, Dayanand Medical College & Hospital,Ludhiana, Punjab Objective:The present study was aimed to find the predictive value by comparing the lactic acid level in the peritoneal fluid and is the blood of patients with acute abdomen. Methods:Lactic acid was estimated in peritoneal fluid and blood in fifty patients with acute abdominal conditions blood lactic acid in patients with surgical abdomen was 3.85+2.96 mmol/l as compared to 4.36+4.34 mmol/l in patients with non-surgical abdomen. The peritoneal fluid lactic acid levels were significantly higher in patients with gangrenous gut, perforation of hollow viscous and secondary peritonitis. There was no significant difference in blood lactic acid in two groups. Peritoneal fluid lactic acid levels were significantly higher than the blood lactic acid in patients with surgical abdomen.

(30 patients with surgical abdomen and 20 patients with non-surgical abdomen) by Blood Gas Analyzer. The mean peritoneal fluid lactic acid values in patients with surgical abdomen was Results: The determination of peritoneal fluid 14.65+6.54 mmol/l and in patients with non- lactic acid concentration particularly when surgical abdomen was 5.92+4.34 mmol/l and the compared with blood lactic acid levels, is

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potentially useful in the patients who presented is most beneficial when the patient's neurologic with acute intra-abdominal pathology. status or unresponsiveness hampers the standard clinical evaluation and in the clinically ill medical Conclusion:Although its usefulness would be patients in whom a negative laparatomy might in limited in patients with clear clinical indications itself be catastrophic. for urgent laparatomy, this diagnostic maneuver Abstract No: 13.16

Estimation of Serum Enzymes in Antibiotic Therapy for Assessment of Hepatotoxicity

Sen S*, Sinha S**,Nazmi A ***,Tripathi S**** and Prashant**** Dept of Pharmacology & Biochemistry,Index Medical College,Indore, Madhya Pradesh, *Asst. Professor, Dept. of Pharmacology;**Demonstrator,Dept of Biochemistry,***Asst. Prof;**** Asso. Prof,Dept of Pharmacology,Index Medical College,Indore,MP-452016. Objective: To elucidate the effect of antibiotics that are causing hepatotoxicity and to assess the relationship between parameters in the patients of antibiotic therapy. Methods: Blood samples were collected after overnight fasting for 12 hrs and serum were separated within one hour in admitted patients who were on antibiotic therapy for more than 14 days in various departments of Index Medical College Hospital & Research Centre, Indore. Exclusion criteria: patients with preexisting liver disease, chronic alcoholics, history of acute hypotension, malaria parasite toxicity were excluded from this study. Based on activity of transaminase enzymes the patients were divided into two groups. 42 patients who developed hepatotoxicity were in Group 1 and remaining 35 patients who did not Abstract No: 14.1 report heptotoxicity were in Group2. Parameters (AST, ALT, ALP & LDH) were estimated in semi auto analyzer (EBRA) using suitable standard methods & correlated with normal reference values using suitable statistical method (`t' test). Results: Group1 patients(n=42) shows antibiotic toxicity with mean values were AST 100.8± SD 66.79, ALT 117.6± SD 110.7, ALP 154.4±SD 42.48 ,LDH 178±45.9.Whereas group2 patients(n=35) with no hepatotoxicity have shown mean values of AST 36.51± SD 7.35, ALT 40.7± SD 6.42, ALP 122.5±SD 13.4 ,LDH 144±38.1. Conclusion: Present study was done to assess antibiotic induced hepatotoxicity by measurements of the serum liver enzymes and necessity of dose adjustment among those patients.

Phytochemical Investigation and Anthelmintic Activity of Extracts of Seeds of Elletaria cardamomum

A.A Bidkar, M. K. Aswar*, K.N. Gujar and C. S. Patel Sinhgad Institute of Pharmacy, Narhe, Pune-411041(M.S) Email ID: [email protected], [email protected] Background: Elletaria cardamomum is a widely with wide ranging biological activities. It is used acclaimed acclaimed Ayurvedic medicinal plant in treating various gastrointestinal, cardiovascular

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and neuronal disorders. It is attributed with antimicrobial activity, gut modulatory, blood pressure lowering, diuretic and sedative activities. A literature survey revealed that the anthelmintic activity has not been evaluated so far hence Objective: of the present work was to evaluate these activities of the extracts of the seeds of the plant Methods: The coarse powder of the seeds of E.cardamomum were defatted with petroleum ether and subjected to continuous hot water extraction using with methanol and water. The extracts were filtered and concentrated under reduced pressure to obtain the extracts as solid Abstract No: 14.2

residues. Preliminary Phytochemical screening of extracts revealed the presence of Flavanoids, alkaloids, fats & oils and tannins. Both the extracts were subjected to screening for anthelmintic activity on earthworms Results; The methanolic extract showed a significant anthelmintic activity as compared to albendazole and aqueous extract. Conclusion: Hence from the results of Phytochemical and pharmacological; investigation it can be concluded that, methanolic extract shows more significant anthelmintic activity compared to aqueous extract and Albendazole.

Phytochemical Investigation and Pharmacological Evaluation of Extracts of Leaves of Calotropis procera Linn.

A.A Bidkar, K.N. Gujar, U.S. Bagul* K.S.Kanugo and K.J. Oza Sinhgad Institute of Pharmacy, Narhe, Pune-411041(M.S), Email ID: [email protected], [email protected] Background:Calotropis procera (R.Br) is a widely acclaimed Ayurvedic medicinal shrub with wide ranging biological activities. It is used generally for its emetic, anti-inflammatory, diuretic, anthelmintic, analgesic well as hepatoprotective properties. Very commonly used in asthma, piles and skin diseases. presence of Flavanoids, Steroids and tannins. Both the extracts were subjected to screening for locomotor activity and anxiolytic activity.

Results: Locomotor activity and anxiolytic activity was ascertained on white albino mice using Actophotometer model and elevated plus maize model respectively. The alcoholic extract showed Objective:A literature survey revealed that the the depression in locomotor activity, due to CNS controlled locomotor activity and anxiolytic increase in the GABA concentration in the brain, activity has not been evaluated so far. Hence more than the Diazepam and aqueous extract. objective of the present work was to evaluate these Similarly the alcoholic extract showed greater activities of the extracts of the leaves of the plant. anxiolytic activity as compared to aqueous extract and Diazepam Methods: The shade dried leaves of C.procera were defatted with petroleum ether and subjected Conclusion:Hence from the results of to continuous hot water extraction using with phytochemical and pharmacological investigation it can be concluded that, alcoholic extract shows alcohol (95% v/v) and water. The extracts were more significant CNS depressant activity in the filtered and concentrated under reduced pressure form of locomotor activity as well as anxiolytic to obtain the extracts as solid residues. Preliminary activity compared to aqueous extract and Phytochemical screening of extracts revealed the Diazepam.

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Abstract No: 14.3

Phytochemical Investigations and Anticonvulsant Activity of Seeds of Brassica campestris.

A. Bidkar*, K. N. Gujar, P. S. Parmar, A. A. Deshpande and A.V. Vaidya Department of Pharmacognosy*, STES's Sinhgad Institute of Pharmacy, Narhe, Pune- 411 041 (MS) India, Email ID: [email protected], [email protected] Objective: Pentylenetetarazole (PTZ) induced and duration of PTZ induced seizures in more than seizures in albino mice were used to study 50% of mice. anticonvulsant activity of aqueous, chloroform and alcoholic extract of seeds of Brassica campestris. Results: The standard anti-epileptic drugs (diazepam, phenytoin) however, significantly Methods An aqueous, alcoholic and chloroform antagonized seizures induced by PTZ. The extracts were administered intra-peritoneally (i.p) alcoholic extract significantly depressed the central at doses of 250mg/kg in the experimental model nervous system. The pharmacognostic evaluation and the effects were compared against PTZ of seeds of Brassica campestris was also carried induced seizures in standard, control, respectively. out. Preliminary phytochemical studies revealed Amongst these extracts the alcoholic extract of the presence of alkaloids, flavonoids and tannins. seeds of Brassica campestris at dose of 250mg/ kg showed significant protection against PTZ Conclusion:The results obtained in this study induced seizures. The extract delayed the onset indicate the presence of anticonvulsant activity of alcoholic extract of seeds of Brassica campestris. Abstract No: 14.4

Anticarcinogenic Effect of Indigofera Aspalathoides by Influencing Xenobiotic Enzyme Activity During DMBA Induced Hamster Buccal Pouch Carcinogenesis

Philips Abraham, V. Arul and Dr.S.Sethupathy Animals were sacrificed after 14 weeks. Phase I (Cytochrome p450, Cytochrome b5 and Cytochrome b5 reductase) and phase II (Glulathione-s-transferase, Quinone reductase and UDP-glucuronyl transferase,)Xenobiotic enzyme activity of oral tissues were assayed. Tissue expression of Cytochrome p450B1(CYP1B1),Aryl hydrocarbon receptor(AHR) and Aryl hydrocarbon nuclear translocator (AHNT) were Methods: Syrian Golden hamsters were divided analyzed by RT-PCR. in to 4 groups. The right buccal pouch of animals in Group I & II were painted with 0.5% DMBA Results: All hamsters painted with DMBA alone 3 times a week for 14 weeks. Group II also developed buccal pouch cancer. Phase I enzyme received an oral dose of ethanol extract of IA (300 activity was observed to be elevated and phase II mg/Kg body wt). Group III was treated with the enzyme activity was found to be decreased in this extract alone. Group IV served as the control. group. Administration of IA extract has prevented Objective: Research for new drugs in the field of Oncology has been refocused on natural products -particularly from the plants that is being used as Folk medicine. This study evaluated the anticarcinogenic effect of extract from the leaves of Indigofera Aspalathoides(IA) - which is traditionally used to treat various skin disorders and cancer.

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carcinogenesis with a significant decrease in phase I enzyme and an increase in phase II enzyme activities.

Conclusion:From the above observation, we propose that influence of IA extract on xenobiotic enzyme activity may be considered as one of the factor for its anticarinogenic property.

Abstract No: 14.5

Biochemical Changes of Feeding Nigella sativa to Rats Fed a High Fat Diet and Treated with Adrenaline

Bini Balakrishnan, Ceema Varghese, Nithi Varghese, Vidya A B, Praveen K V, Sheeba Varghese, J K Mukkadan and P K Joseph Little Flower Institute of Medical Sciences and Research, Angamaly 683 572, India. Email ID: [email protected] and moderate reduction in the levels of blood glucose and liver lipids. When nigella was fed along with HFD there was reduction in liver and serum lipids, serum AST, ALT and ALP and blood glucose. When rats fed HFD and nigella was Results: Rats fed a diet containing 40% dalda treated with the hormone, the adrenalin induced (HFD) exhibited higher levels of total cholesterol rise in serum enzymes was much less. Also the and triacylglycerol in liver and serum, liver levels of serum and liver lipids were less than in thiobarbituricacid reactive substances (TBARS), HFD fed rats in spite of adrenaline treatment. serum levels of AST, ALT, and ALP and reduced levels of serum albumin. When HFD fed rats were Conclusion: Nigella sativa is cardioprotective and treated with 2 mg adrenaline per Kg, there was hepatoprotective. phenomenal increase in serum AST, ALT and ALP Objective: Nigella is a plant used in ayurveda and its seeds are considered to be cardioprotective and hepato-protective. To study this, the present study was undertaken. Abstract No: 14.6

Biochemical Changes of Feeding Desmodium gangeticum to Rats Fed a High Fat Diet and Treated with Adrenaline

Ceema Varghese, Bini Balakrishnan, Nithi Varghese, Vidya A B, Praveen K V, Sheeba Varghese, J K Mukkadan and P K Joseph Little Flower Institute of Medical Sciences and Research, Angamaly 683 572, India; Email ID: [email protected] Objective: Desmodium is a plant used in ayurveda and extracts of its roots are considered to be cardioprotective and hepatoprotective. To study this, the present study was undertaken. Results: Rats fed a diet containing 40% dalda (HFD) exhibited higher levels of total cholesterol and triacylglycerol in liver and serum , liver

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thiobarbituricacid reactive substances (TBARS), serum levels of AST, ALT and ALP, and reduced levels of serum albumin. When HFD fed rats were treated with 2 mg adrenaline per Kg, there was phenomenal increase in serum AST, ALT and ALP and moderate reduction in the levels of blood glucose and liver lipids. When desmodium was fed along with HFD there was reduction in liver

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and serum lipids, serum AST, ALT and ALP, and blood glucose. When rats fed HFD and desmodium were treated with the hormone, the adrenaline induced rise in serum enzymes was much less. Also the levels of serum and liver lipids Abstract No: 14.7

were less than in HFD fed rats in spite of adrenaline treatment. Conclusion: Desmodium gangeticum is cardioprotective and hepatoprotective

Comparative Study of Jamun Powder and Tulsi Leaf Powder as Hypoglycemic and Hypolipoidemic Agent in Diabetes Mellitus Type 2

Dr. Meena Verma1, Deependra Sharma,1 Dr. Sangeeta Paneri1 and Dr.A.Mishra2 1 Dept. of Biochemistry M.G.M. Medical College Indore. 2 Dept. of Biochemistry SAIMS Medical College Indore Objective: A study was done to evaluate the Results: The comparative study has shown hypoglycemic and hypolipoidemic effects of significant value in between control and study group. After comparison of both supplements jamun seed powder and tulsi leaf powder. jamun powder and tulsi leaf powder in study group, Methods: Among the 60 obese and diabetic jamun powder have shown highly significant value subject of 35 years to 55 years. 10 were taken as in serum glucose and significant in serum lipids, healthy control and 25 diabetic and obese were but the tulsi leaf powder has shown significant treated with jamun seed powder and 25 were values in serum glucose and non significant in treated with tulsi leaf powder. The criteria of serum lipids as compared to jamun seed powder inclusion of study group in subjects was FBS > group. 110 mg/dl of glucose, >290 mg/dl of cholesterol, > 210 mg/dl of triglyceride, <30 mg/dl of HDL, Conclusion: This study shows that jamun seed and > than 180 mg/dl of LDL. All the subjects powder is more effective in lowering of serum glucose were examined for serum glucose and serum lipids. as well serum lipids than the tulsi leaf powder. Abstract No: 14.8

Nyctanthes arbortristis Extract Restores the Cartilage Damage During Experimental Arthritis

Brijesh Rathore, A A Mahdi, B N Paul*, P N Saxena**, Farzana Mahdi# and S K Das## Department of Biochemistry and Rheumatology##, King George's Medical University, Lucknow, # Department of Biochemistry, Era's Lucknow Medical College, Sarfarazganj, Hardoi Road, Lucknow 226 003, *Immunobiology Laboratory, Industrial Toxicology Research Centre, M. G. Marg, Lucknow, **Deparment of Zoology, Dr. B R Ambedkar University, Agra. Objective: Rheumatoid arthritis is an autoimmune disease, characterised by progressive cartilage and bone destruction. Normal synovium is a delicate tissue, lining the joint capsule but in arthritic condition, the synovium transforms into an aggressive tumor like structure called pannus, which invades and erodes the joint. In later stages, the presence of macrophage, fibroblasts, mast cells and chondrocyte accumulation is found at the site of cartilage erosion. However, the pathogenesis of disease is unclear but activated synovial fibroblasts appear to play a major role in disease. It is

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speculated that their ability to induce and perpetuate inflammation is associated with the production of pro-inflammatory cytokines which inturn results in inflamed synovium. Methods: Arthritis was induced in mice by injecting the Freund's Complete Adjuvant in the foot pad. A booster dose was given on the day 12 th . Nyctanthes arbor tristis extract was administered orally from the day of induction of disease. Abstract No: 14.9

Results: On the day 47th animals were sacrificed and the ankle joint was processed for the histopathology. Infiltration of inflammatory cells, neovascularisation and cartilage damage was found negligible in the extract treated animals as compared to the arthritic one. The cell lining was also observed intact in Nyctanthes arbor tristis extract treated animals as compared to the control animals. Conclusion:Our findings open up new herbal therapeutic approach for rheumatoid arthritis.

Polyherbal Anti Diabetic Drug: an Approach to Cure Diabetes

Gaikwad SB*, Bhagwat VR* and Trivedi DJ ** * Dept of Biochemistry, SBH: Govt.Medical College, Dhule, ** Dept of Biochemistry, SDN Medical College, Dharwad .Email: [email protected] Background: In the last few years the herbal medicine and drugs are gaining popularity, because of their natural origin and less side effects compared to modern synthetic chemical compounds. To date, over 400 traditional plant treatments for diabetes have been reported, although only a small number of these have received scientific and medical evaluation to assess their efficacy. There are several mechanisms through which these herbs act to control the glucose level, majority of the actions remains unexplored. The purpose of this study was to evaluate the possible hypoglycemic actions of a polyherbal mixture of 17 ingredients which are commonly used as food and in other herbal treatments. Objective: The present study was designed to investigate the effect of a polyherbal drug, on blood glucose, glycosylated haemoglobin and insulin level in alloxan-induced diabetic rats. Group 1: normal rats given 2 ml of saline; group 2: normal rats given aqueous solution of drug (0.25 g/kg of body weight); group 3: diabetic control rats given 2 ml of saline; group 4: diabetic rats given aqueous solution of drug (0.05 g/kg of body weight); group 5: diabetic rats given aqueous solution of drug (0.10 g/kg of body weight); group 6: diabetic rats given aqueous solution of drug (0.20 g/kg of body weight) and group 7: diabetic rats given aqueous solution of Metformin (500 µg/ kg of body weight). The treatment was given for 30 days. After the treatment, fasting blood glucose, plasma insulin and glycosylated haemoglobin were determined in normal and experimental animals. Results: Treatment with drug resulted in a significant reduction in blood glucose, glycosylated haemoglobin value, but plasma insulin level did not show any significant change.

Conclusions: The present investigation indicated that the polyherbal drug has shown significant Methods: Male Wistar rats, body weight of 160- control on the blood glucose level. The herbal 200 g (10 normal and 25 diabetic rats), were used combination could be decreasing the insulin in this study. The rats were divided into seven resistance thereby increasing the uptake of glucose groups after the induction of alloxan diabetes. In by cells, resulting into lowering of the glycosylated the experiment, five rats were used in each group. haemoglobin value and blood sugar level.

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Abstract No: 14.10

Extraction and in vitro Evaluation of Biological Activity of Crude Polyphenolic Extracts from Indian Grapes (Vitis vinifera) on Transformed (AW13516) and Nontransformed (HaCaT) Stratified Epithelial Cells.

Neha Tamhankar (a), Geeta Ibrahim (a), and Milind Vaidya (b) (a) College of Home Science, Nirmala Niketan, New Marine Lines, Mumbai-40002 (b) ACTREC (Advanced Centre for Research and Education in Cancer), Kharghar, Objective: Cancer being one of the leading causes of mortality in the world, preventive measure is the most cost effective long term strategy for combating the disease. Recently, naturally occurring polyphenolic antioxidants obtained from fresh fruits and vegetables have been identified as potential chemopreventive agents. Due to lack of substantial data about crude polyphenolic extracts from Indian grapes, this study was carried out to evaluate the biological activity of the extracted polyphenols on stratified epithelial cell lines (HaCaT and AW-13516). Methods: The grape varieties selected for the study were namely Pandhari sahebi (PS) and Thompson seedless (TS). The crude extracts were analysed for their polyphenol content as well as antioxidant activity using gallic acid as the standard. Abstract No: 14.11 Results: The polyphenolic content of both the varieties were around 1.5mgs gallic acid / gm freeze dried powder but the seed showed a very high polyphenol content and IC50 value was 78µg/ ml. Sulphorhodamine B assay was carried out on the transformed (AW-13516) and non transformed (HaCaT) cell lines and it was found that PS polyphenolic extract was more effective as compared to the TS polyphenolic extract in causing cell growth inhibition. Also the effect seen was greater on the transformed cell line as compared to the non - transformed. A rapid assessment survey was undertaken in order to know the awareness of grape polyphenols among consumers and it was seen that about 69% of the consumers were unaware of polyphenols and hence there is a need to spread awareness regarding the same.

Beneficial Effects of Tetrahydrocurcumin and Chlorogenic Acid on Lipid Profile in Streptozotocin-Nicotinamide-Induced Diabetic Rats

Karthikesan. K and Pari. L Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalainagar-608 002, Tamilnadu, India. Objective: The present study investigated the effect of tetrahydro curcumin (THC) and chlorogenic acid (CGA) on lipids, lipoproteins and lipid metabolizing enzymes in streptozotocin (STZ)-nicotinamide (NA)-induced diabetic rats were examined. intraperitoneal (i.p) injection of STZ (45 mg/kg b.w) 15 minutes after the i.p administration of NA (110 mg/kg b.w). Diabetic rats showed a significant (p<0.05) increase in the levels of lipids (cholesterol, triglycerides (TGs), free fatty acids (FFAs) and phospholipids (PLs)) in plasma and tissues (liver and kidney).

Methods: Diabetes was induced in experimental rats weighing 180­220 g, by a single Results: The plasma levels of low-density and very

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low-density lipoproteins (LDL and VLDL respectively) were increased, whereas the levels of high-density lipoproteins (HDL) were significantly decreased (p<0.05). The activity of 3-hydroxy 3-methylglutaryl Coenzyme A (HMG CoA) reductase was increased significantly (p<0.05) in liver and kidney and the activities of lipoprotein lipase (LPL) and lecithin cholesterol acyl transferase (LCAT) decreased significantly (p<0.05) in the plasma of diabetic rats. Oral treatment of THC (80 mg/kg b.w) and CGA (5 mg/kg b.w) to diabetic rats for 45 days produced

a significant (p< 0.05) decrease the concentration of plasma and tissue lipids. Lipoprotein levels were restored to near normal levels. The activity of HMG CoA reductase decreased in the tissues and the activities of plasma LPL and LCAT increased significantly (p<0.05) in THC and CGA treated diabetic rats. Conclusion: These results showed that THC and CGA supplements seem to be beneficial for correcting the lipid profile and preventing diabetic complications.

Abstract No: 14.12

Search for a Perfect Diet Including Antioxidants

Prof (Dr) Kaushal K.Srivastava* 12/4B, Aravali View Rail Vihar, Sector56, Gurgaon (Haryana) *Advisor and Consultant, Pharma Industries, President, DelhiChapter ACBI, Emeritus Professor, ACBR, Delhi University, Delhi-110007. Email: [email protected] Diet pattern of a population depends largely on the traditions, availability of food items, agricultural and fruiticultural output. Due to globalization, easy storage and transportation the products from one corner of the world are available all over. Yet, the search for a perfect diet continues especially with reference to promotion of health and prevention of chronic life style diseases such as cardiovascular diseases, cerebral strokes, neurological disorders, diabetes and even some type of cancers. A large number of studies have been made on the role of various food components like simple and complex carbohydrates, saturated, unsaturated fats, and fibers in food, minerals and water in connection with the incidence of chronic life style diseases. The results have lead to confusing recommendations from time to time. The role of free radicals induced damage and its prevention by antioxidants has been widely

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described in these conditions. Sometimes increased intake of antioxidants has resulted in increased incidence of disease which it intended to prevent such as carotene and smoking induced lung cancer. More often, the intake of antioxidant supplements such as Vitamin E, C and Polyphenols including Flavanoids had little effect.The search for a perfect diet continues unabated to promote health and prevent ageing and disease. References: Jonathan Scheff (2008) Scintific American June 6. Trivedi,Bijal (2006) New Scientist. September 23, 42-47 The Good, The Fad, The Unhealthy Martin, Lisa (2006) New Scientist. August 5, 40-47 The antioxidant myth

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Abstract No: 14.13

Antioxidant Evaluation of Some Hepatoprotective Medicinal Plants

Chorge Ketan, , Dr. (Mrs.) Kale K. U. and Chorge Ulka The Institute of Science, 15 Madam Cama Road, Fort, Mumbai-32. Email: [email protected] Objective: Hepatitis is well known common liver dysfunction all over the world. There are different types of jaundice like Infectious hepatitis, Obstructive jaundice, Alcoholic hepatitis, Malerial hepatitis; Drug induced jaundice, Cirrhosis of liver, Viral hepatitis (A to G). It is generally due to long-term alcohol abuse, infection, or exposure to various chemicals and drugs. A condition characterized by yellowness of skin and eyes, mucous membranes and body fluids due to deposition of bile pigment resulting from excess bilirubin in blood (hyperbilirubinanemia). Some allopathic medicines are known to cure hepatitis. However, their recovery is very slow and having side effects of their own. Literature reviews reveal that the herbal medicines are found to be more effective for different types of liver dysfunction. Hence, herbal medicines provide a wide scope in attempting cure and treatment of liver dysfunction, in general known as jaundice. Active principles Abstract No: 14.14 extracted from plants like phyllanthus niruri, silybum marium etc. are showed to be effective against liver dysfunction. Herbs have pronounced hepatoprotective activity and would be useful tool in controlling and treating jaundice. An attempt was done to do comparative study of extraction, identification, separation and characterization of herbal active principles from medicinal plants and to compare the characteristics. Methods: The present study involves isolation of phyllanthin a metabolite from the plant of phyllanthus niruri and silymarin from silybum marium and comparison of antioxidant activity of the total extract with the isolated compounds. The antioxidant activity was compared spectrophotometrically using DPPH. Results: The results reveal good antioxidant activity as compared to standard used ascorbic acid.

Does Chilli Affect Platelet Aggregation?

Kiran DK Ahuja, Murray J Adams, Dominic P Geraghty and Madeleine J Ball School of Human Life Sciences, University of Tasmania, Launceston, Australia [email protected] Objective: Two separate studies were conducted absence and presence of capsaicin and to investigate the concentration/dose effect of dihydrocapsaicin (final concentration, 3.125 ­ 100 chilli (capsaicin) on platelet aggregation under in µmol/L). vitro and ex vivo conditions. Study 2: Fourteen participants undertook five Methods: Study 1: Venous whole blood was meal-challenge tests, each in a random order. collected from six healthy subjects (aged 23-39 Meals comprised of bread, meat patty and a years) into 3.2% sodium citrate containing glucose drink. Three of the five meals also anticoagulant tubes. Platelet rich plasma (PRP) contained 20, 30 and 40g of chilli paste, was separated and aggregation induced with respectively. One meal included capsules filled adenosine diphosphate (ADP; 5 µmol/L) in the with dry chilli powder. The other meal was a

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placebo. PRP separated from citrated plasma collected at fasting and 40 and 120min postprandially was subjected to ADP (2.5, 5 and 10 µmol/L) induced platelet aggregation.

Results: Study 1: Concentration-dependent decreases in ADP-induced platelet aggregation were observed with both capsaicin and dihydrocapsaicin (0 vs 100 µmol/L) for max % Conclusion: Chilli (capsaicin) helps inhibit aggregation and % area-under-curve (all p<0.05). platelet aggregation, a risk factor for cardiovascular disease. Abstract No: 14.15

Study 2: Intake of all five meals resulted in a reduced maximum platelet aggregation at 120min postprandially compared to the fasting samples (p<0.05). The three meals containing chilli paste resulted in further inhibition of platelet aggregation compared to the bland (placebo) and the chilli capsule meals.

Antidiabetic Effects of the Aqueous Extract of Fruit of Withania Coagulans in Diabetic Albino Rats

Kirtikar Shukla, JK Gambhir and Rimi Shukla Department of Biochemistry, University College of Medical Sciences and G.T.B. Hospital, Shahdara, Delhi-110095; E.mail: [email protected] Objective: Diabetes mellitus is a clinical syndrome characterized by a common phenotype of hyperglycemia and dyslipidemia with subsequent derangement of carbohydrate, Lipid and protein metabolism. Fruit of Withania coagulans is used in the traditional Indian system of medicine for the treatment of diabetes mellitus. Our aim was to investigate the hypoglycemic and hypolipidemic effects of aqueous extract of fruits of Withania coagulans in alloxan induced diabetes mellitus in rats. Methods: Diabetes was induced by intraperitoneal injection of alloxan at dose of 100mg/kg b.wt. Rats weighing 150-200gm were divided into three groups of six each, Group I healthy control, Group II diabetic control and Group III Diabetic rats treated with Withania coagulans at dose of 250 mg/kg b.wt./day orally for 30 days.

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Results: The Group II, rats showed increased fasting & postprandial blood sugar, serum cholesterol, triacylglycerol levels and decreased level of HDL as compared to Group I. Treatment with water extract of Withania coagulans, significantly decreased fasting and postprandial glucose levels in group III as compared to group II diabetic control rats (p<0.05). Serum cholesterol and triacylglycerol levels also decreased significantly in diabetic treated rats as compared to group II (diabetic rats) (p<0.05), while HDL level increased as compared to Group II diabetic control rats (p<0.05). Glycosylated hemoglobin (HbA1c) was 8.79 in diabetic group which decreased to 5.9 in WC treated group While GHb A1c of healthy control was 4.65. Conclusion: These results show that water extract of fruit of Withania coagulans has potential to be used in the treatment of diabetes mellitus.

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Abstract No: 14.16

A Putative Anticancer Mechanism of Plant Derived Polyphenols

Mohd. Fahad Ullah, Uzma Shamim, Sarmad Hanif, Asfar S Azmi, Showket H Bhat and S.M.Hadi Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh 202002, India; Email : [email protected] Nutraceuticals are defined as food or parts of food that provide medical or health benefits including the prevention or treatment of diseases. Many plant derived foods and beverages contain the class of nutraceuticals known as polyphenols. As such plant polyphenols are important constituents of human diet and a number of these are considered to possess chemopreventive and therapeutic properties against cancer. They are recognized as naturally occurring antioxidants but also act as prooxidants catalyzing oxidative DNA degradation in the presence of transition metal ions such as copper. Using human peripheral lymphocytes and Comet assay we have shown that polyphenols are capable of causing cellular DNA breakage. Incubation of lymphocytes with neocuproine (a cell membrane permeable Cu (I) chelator) inhibited DNA degradation confirming that Cu (I) is an intermediate in the DNA cleavage reaction. Bathocuproine, which is unable to Abstract No: 14.17 permeate through the cell membrane, did not cause such inhibition. Further we have shown that polyphenols are able to degrade DNA in cell nuclei and that such DNA degradation is inhibited by neocuproine as well as bathocuproine (both of which are able to permeate the nuclear pore complex) but not by compounds that specifically bind iron and zinc (desferrioxamine mesylate and histidine respectively). Copper is a major metal ion present in nuclei and it is well established that tissue, cellular and serum copper levels are considerably elevated in various malignancies. Therefore cancer cells may be more subject to electron transfer between copper ions and polyphenols to generate reactive oxygen species. Such a common mechanism would better explain the anticancer effects of polyphenols with diverse chemical structures as also the preferential cytotoxicity towards cancer cells

A Comparative Study of Nitric Oxide and Hydrogen Peroxide Scavenging Activities of Botanical Extracts Ocimum sanctum and Wagatea spicata.

Geetha Samak1, Revathi P Shenoy 2 and D. M. Vasudevan 3 1 DVS College of Arts and Science, Shimoga, Pin 577201, Karnataka, India , 2 Department of Biochemistry, Kasturba Medical College, Manipal University, Manipal Pin 576104 Karnataka, , 3 Professor of Biochemistry and Chairman Amrita laboratory service, Amrita school of Medicine, Amrita Vishwa Vidyapeetham, Cochi -682041, Kerala. Objective: Ocimum sanctum, a traditional medicinal plant of India, showed significant abilities of scavenging superoxide, hydrogen peroxide radicals and inhibiting lipid peroxidation. Wagatea spicata, a medicinal plant found in Western Ghats of India is also potent scavenger of highly reactive superoxide and hydroxyl radicals. Methods: In the present study we evaluated Nitric oxide(NO) and Hydrogen Peroxide(H 2 O 2 ) scavenging abilities of the plants in comparison

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with known antioxidant Ascorbic acid. Shade dried plant material powder was extracted with water, fractionated with different solvents. Total phenolic and flavonoid content of the plant materials were analyzed through spectrophotometry. NO was generated from sodium nitroprusside and scavenging action was measured by the Greiss reduction. H2O2 scavenging action was measured by peroxidase based assay system. Results: Botanical extracts of both the plants showed concentration-dependant inhibition of H2O2 and NO. Ocimum leaves showed better scavenging of H2O2 than Wagatea. Crude extract of Ocimum showed IC50 at 18.5 ± 6.2µg/ml, ethyl acetate and diethyl fractions having IC50 at Abstract No: 14.18

5 ± 1.4µg /ml, 6.5 ± 2.9µg /ml respectively which is almost equal to that of ascorbate. Wagatea showed significant NO scavenging action than ocimum. Flower and bark extracts, their methanol and water fractions of Wagatea showed NO scavenging activity having IC50 at 10 ± 7µg/ml. Free radical scavenging action of Wagatea spicata and Ocimum sanctum can be correlated with its phenolic and flavonoid rich plant materials. Conclusion: These results indicate that both these plants can scavenge deleterious NO and H2O2 radicals at low concentration. Phenolic and flavonoid rich Ocimum and Wagatea can be supplemented as antioxidant equivalent to ascorbate.

Cardioprotective Effect of Terminalia arjuna on Isoproterenol Induced Myocardial Infarction in Rats

S.K. Shukla*, S. Dwivedi**, U.R. Singh*** and S.B. Sharma* Department of Biochemistry*, Department of Medicine and Preventive Cardiology**, Department of Pathology***, University College of Medical Sciences (University of Delhi), Delhi -110095. Objective: Cardiovascular diseases are one of the leading cause of morbidity and mortality in developed as well as in developing countries, therefore its prevention is a major public health challenge. Oxidative stress plays a potential role in CAD and it has been reported to accelerates atherosclerosis. Terminalia arjuna is traditionally used for the treatment of heart diseases. Methods: The present study was undertaken to demonstrate the effect of hydroalcoholic extract of Terminalia arjuna bark on lipid peroxidation, reduced glutathione and superoxide dismutase to evaluate the antioxidant activity in experimentally induced myocardial infarction in rats and then sacrificed for histopathological investigation.24 male albino rats weighing 200-250g were divided into three groups (n=8 in each groups).Healthy control (groupI) were given normal saline for 30 days. Isoproproterenol treated rats(groupII) was given normal saline for30 days, Isoproterenol

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(85mg/kg b.w) administered Subcutaneously on 29th and 30th day. To drug treated rats (groupIII) hydroalcoholic extract of T. arjuna (HETA), orally administered for 30 days at a dose of 100 mg/kg b.w, Isoproterenol administered SC on 29th and 30th day. Fasting blood samples were taken prior to and after given the treatment to estimate MDA, GSH and SOD, and then sacrificed for histopathological examination. Results: HETA produced significant depletion in MDA with a concomitant elevation in activity of SOD. GSH level was also significantly increased by HETA (p< 0.001).Histopathological studies showed area of necrosis in isoproterenol treated rats. Conclusion: The results suggest that crude bark of TA augments endogenous antioxidant compounds of rat heart and also prevents oxidative stress associated with Isoproterenol induced ischemia.

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Abstract No: 14.19

Aluminum Induced Changes in Blood and Brain; Protective Role of Bacopa monnieri

Sandeep Tripathi1, Abbas Ali Mahdi1, Mahdi Hasan2, Deepa Prajapati3, Sanjay Annarao3, Raja Roy3 and C. L. Khetrapal3 Departments of 1Biochemistry and 2Anatomy, C.S.M. Medical University, 3Centre of Biomedical Magnetic Resonance, SGPGIMS Campus, Lucknow -226014. Objective: Comparative therapeutic efficacy of Bacopa monnieri extract (BR) and anticholinergic drug donepezil (DP) on aluminum (Al) neurotoxicity have been investigated using Magnetic Resonance Imaging - Magnetization Transfer Contrast (MRI-MTC) imaging, correlative behavioral, biochemical and ultra structural investigations of rat brain followed by metabolic changes in serum and urine and blood biochemistry. Methods: In the present study, male albino rats were administered with AlCl3 (100 mg / kg body weight) for 90 days and simultaneously treated another groups with 40mg and 2.5 mg / kg body weight with Bacopa monnieri and Donepezil respectively. Serum and urine was used to asses the markers of metabolic disturbances followed by the behavioral and MRI-MTC imaging of rats. Furthermore, ultrastructural changes were also evaluated in the hippocampus. Abstract No: 14.20 Results: Large accumulation of age pigment lipofuscin granules, macroautophagy in hippocampus of Al3+ treated rats as observed in the Electron microscopy (EM) images indicates early aging and its similarities with Alzheimer disease, which reflects its direct correlation with lower MTR value. Significantly elevated value of MTR following 90 days co-administration of Brahmi when compared with Al3+ and DP treated rats. Conclusion: On the basis of the results of the present study, we conclude that Al may be responsible for neurolipofuscinogenesis and alteration in neurobehavioral activity and these changes may be responsible for the development of age related diseases like Alzheimer's disease and Bacopa monnieri might be used as neuroprotective agent.

Hypolipidemic Activity of Hibiscus rosa sinensis (root) in Hyperlipidemic Rats

Vishnu Kumar 1, Ranjana Singh 3, Sushma Singh 3, Shivani Pandey 3, Ashok Kumar Khanna 2, Ramesh Chander 1,Pradyumn Singh1, Farzana Mahdi 1, Jitendra Kumar Saxena 2, Vinod Kumar Singh 3 and Raj Kumar Singh 3 1 Department of Biochemistry & Pathology, Era's Lucknow Medical College &Hospital, Lucknow226003, ,2 Division of Biochemistry, Central Drug Research Institute, Lucknow-226001, and 3 Department of Biochemistry, Chhatrapati Shahuji Maharaj Medical University ( Formerly King George's Medical University ), Lucknow- 226003. The hypolipidemic activity of Hibiscus rosa extract has been studied in triton and cholesterol sinensis (family Malvaceae; Hindi name Gudhal, rich high fat diet (HFD) fed hyperlipidemic rats. Japa in Sanskrit and shoe flower in English) root In triton WR-1339 induced hyperlipidemia,

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feeding with root extract (500mg kg-1 b.w.) exerted lipid lowering effect as assessed by reversal of plasma levels of total cholesterol:TC (22 %), phospholipids:PL (19 %) and triglyceride:TG (22 %) following reactivation of Post Heparin Lipolytic Activity:PHLA (11 %). In another model, chronic feeding of this natural product (500mg kg-1 b.w.) in animals simultaneously fed with high fat diet for 30 days caused lowering of lipid levels in plasma TC (20 %), PL (23 %), TG (20 %), reactivation of PHLA (16 %) and in liver homogenate decrease in TC (21 %), PL (23 %), TG (24 %) with stimulation of hepatic lipolytic activity 17 %. The hypolipidemic activity of Hibiscus rosa sinensis root was compared with a standard drug guggulipid (200mg kg-1 b.w.), a known lipid lowering drug in both models. Histopathology of liver in rats fed with Abstract No: 14.21

cholesterol rich diet showed microvesicular steatosis and occasional macrovesicular in zone 3 (perivenular area) of hepatic lobules along with sinusoidal dilation and mild congestion and the portal trials revealed mild increased in inflammatory infiltrate comprising of mononuclear cells and neutrophils. The liver histopathology in rats which simultaneously also received Hibiscus root extract showed only mild feathery degenerative changes and mild vascular congestion but no steatosis Kupfer cell linings the sinusoids were prominent and increased in number. Their portal triads showed increased inflammatory infiltrate rich in mononuclear cells admixed with polymorphs and fair number of eosinophils. The overall hepatic architecture was maintained.

Cholinesterase Activity in Health Workers Involved in Handling and Spraying of Organophosphorous Insecticides

Himanshu Madaan, Veena Singh Ghalaut and Ashuma Sachdeva Deptt of Biochemistry, Pt. B D Sharma Univ of Health Sciences, Rohtak, Haryana, [email protected] ; [email protected] Background: Anyone exposed to Organophosphorous (OP) pesticides can develop lowered cholinesterase levels; therefore regular checking of their AChE levels should be done. Nearly all the insecticides have the potential to significantly alter the ecosystem. In humans, they are responsible for acute poisonings as well as for long term health effects, Organophosphates and Carbamates are, nowadays, widely used as insecticides. These compounds can cause toxicity through all routes of exposure-dermal, oral, respiratory and, even, ocular. They exert their effect mainly by the inhibition of Acetylcholinesterase (AChE), an enzyme that

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occurs in the CNS, which functions by removing acetylcholine (ACh) from its postsynaptic receptors. Methods: The present study was conducted on 38 subjects working as Multipurpose Health Workers (MPHW GROUP-I) and Field Workers (FW GROUP-II) in District Rohtak, Haryana India to see the effect of exposure to OP insecticides. Serum cholinesterase activity was measured by a kinetic method based on hydrolysis of butrylthiocholine. Results: The MPHW were involved in storing and distributing of the insecticides, whereas the FW were involved in the actual spraying of the insecticides. The mean cholinesterase activity in

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group- 2 (5410.56+957.66) was lower by 27.76% reduction in the levels of PChE, compared to the workers involved in only the handling of the than that in group-1 (7489.28+1106.95). insecticides indicating that this group is at a bigger Conclusions: The workers involved in actual risk. So, the improved controls on workplace spraying of the insecticides had a more marked exposure to pesticides should be focused on these Abstract No: 14.22 individuals.

Lead Poisoning in Residential Areas Near Drainage Systems in Bangalore

Anitha Jerry Juneeja Varghese and Antony.P.U Under the guidance of Dr. Venkatesh. T With the accumulation of lead containing substances in drainage systems, there is a possibility of this lead-contained water to seep into the groundwater and to be used by the residents of the nearby area. In which case, the water might be able unusable. This was checked by a study done in a few areas of Bangalore near large drainage systems. Both bore well and normal well water samples were collected and tested for their Abstract No: 14.23 level of lead concentration. Most results remained within the prescribed limits, except for a few odd samples. The variation in the few samples can be ascribed to the activities in the surrounding areas and certain human errors that could have occurred during the course of the sample collection. This showed that the water used in residential areas near large drainage systems were free from the harmful concentration of lead and was safe for use.

Study of ADA Levels in Tuberculus and Non tuberculus Pleural Effusion

K. Sravanthi, Dr. B. Prabhakar Rao, Dr. V. Sunanda and Dr. C.N. Prasad Prathima Institue Of Medical Sicences, Karimnagar, Andhra Pradesh. E.mail : [email protected] Objective: To assess the diagnostic level of ADA in pleural effusion fluid. Methods: The present study was carried out on 60 patients suffering from pleural effusions of various etiologies. 40 Patients with history and clinical examination suggesting of Tubercular pleural effusion admitted in chest unit of Prathima Institute of Medical Sciences, Karimnagar where included in the study & labeled as group ­ I. In addition as a control of the study, 20 selected patients having Transudate pleural effusions secondary to cirrhosis of liver of Cardiac Heart Failure etc., are taken as group ­ II. This selection criteria was based on history, clinical examination and by fluid protein estimation to differentiate exudates from transudate (Lights Criteria). The ADA activity was compared in both the groups and a cut of value was set as 40 U/L. The results were analyzed by applying "Chi Square Test". Results: The ADA activity of Group ­ I was 40.6 + 9.8 U/L. While that of Group -II was 24 + 10 U/L. The "P" value was < 0.001. The results of the study favors application of ADA activity in Pleural Fluid as the diagnostic tool for Tuberculus pleural effusion without performing any invasive procedures like pleural biopsy.

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Abstract No: 14.24

A Brief Study of Lead Poisoning Among Children

Faculty and Students Dept. Of Biotechnology, NMKRV College for Women, Bangalore; E-mail id: [email protected] Ph.No: 080-26643499 Children are more susceptible to lead poisoning because their bodies are still developing and their body's tend to retain more lead than adults. 30 students from Byramangala government school were tested for blood lead levels. The criteria for choosing Byramangala as the study area was that the untreated domestic sewage of Bangalore city, effluents of various small scale industries are released into Vrishbhavati River which in-turn enters the Byramangala reservoir throughout the year. The water and vegetable samples consumed by these children were analyzed using Atomic Absorption Spectrophotometry. Blood lead levels was analyzed using Anodic Stripping Voltametry using an ESA 3010 lead analyzer. The atomic absorption spectrophotometry showed that the vegetable and water samples had lead levels in "Below Detection Levels" (BDL) and thus may Abstract No.14.25 be ruled out as possible sources of lead poisoning. However the results of the blood analysis obtained remained a cause for concern, as 10% (3 Nos.) were having high blood lead levels, 27% (8 Nos.) were on the borderline, indicating the fact that almost 37% of children have or are on the verge of lead poisoning. The reasons for such high blood lead levels needs to be further investigated and awareness has to be created among the people living in that area. References: a). Chapter 9, AOAC, 18th edition for vegetable samples. b). IS: 3025 (P47) - 1994 for water samples. c) .Anodic Stripping Voltametry using an ESA 3010 lead analyzer.

Iodine Deficiency Disorders Among the Primary School Children of Eastern Nepal.

Gelal B, Chaudhari R, Nepal A, Das BKL, Lamsal M and Baral N* *Professor and HOD, Department of Biochemistry, B. P. Koirala Institute of Health Sciences, Dharan, Nepal . E-mail: [email protected] Objective: To assess the iodine status among the deficient (UIE <100µg/L) children were 26.6% in primary school children of eastern Nepal. Dhankuta and 15.6% in Dharan. In both areas Methods: A population based cross sectional study majority of children consumed packet salt. The was conducted on two schools of Dhankuta and percentages of salt samples with adequately Dharan at January ­ March. A total of 385 urine iodized salt (e"15 ppm) were 83.3% in Dhankuta and equal numbers of salt samples were collected and 89.6% in Dharan. from school children of 6-11 years. UIE was Conclusion: In both areas iodine deficiency is not measured by ammonium-persulphate digestion a major pubic health problem because of normal microplate (APDM) method and salt iodine median UIE, consumption of packet salt and content by semi quantitative rapid test kit. majority of salt samples were adequately iodized. Results: The median UIE of school children of But continuous monitoring program must be Dhankuta and Dharan were 151.13µg/L and continued to ensure the adequate iodine status of 180.34µg/L respectively. The percentage of iodine population

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Abstract No.14.26

Role of Plasma Homocysteine Level in Coronary Artery Disease and Stroke- A Clinical Perspective

*Girish P. Pandey, **Sukhes Mukherjee, **Kannan Vaidyanathan and **D.M Vasudevan *Department of Medicine, **Department of Biochemistry, Amrita School of Medicine, Kochi Objective: Homocysteine is a compound formed in the trans-sulphuration pathway from methionine via the intermediates S Adenosyl methionine and S adenosyl Homocysteine. Homocysteine is commonly associated with various diseases and its elevated levels are known to be associated with cardio-vascular disease, stroke, pre-eclamsia of pregnancy and dementias. Homocysteine concentrations will be estimated by ELISA. Estimations of other enzymes like cystathionine beta synthase, methylene tetrahydrofolate reductase and methoinine synthase was also measured.

Results: There was significant elevation of plasma homocysteine level in acute myocardial infarction patients when compared with control group. There Methods: We studied the total Homocysteine was also alteration of the other enzymes studied. levels in patients selected from our cardiology and neurology department. 95 patients in each Conclusion: Our data from this study provide category of CAD and stroke were compared with evidence that plasma homocysteine levels are 105 normal individuals coming to the medicine elevated in patients with acute myocardial department for routine check up. Total infarction. Abstract No.14.27

Vitamin B12 and Folic Acid Levels in Tobacco Chewers: A Duration Dependent Study.

Indira Samal1, M.Maneesh, Sukhes Mukherjee1 and Subir K Das2 . 1 Dept of Biochemistry, Amrita Institute of Medical Sciences, Kochi, Kerala. 2 Dept of Biochemistry, Govt Medical College, Agartala, Tripura. Background: Cytotoxic effects of chewing tobacco are mediated through production of reactive oxygen species. Superoxide dismutase (SOD) and Glutathione Reductase are antioxidant enzymes.Vitamin B12 and folate are cofactors in the synthesis of DNA.Vitamin B12 is also a cofactor in the synthesis of methionine from homocysteine, increased levels of which are risk factors for coronary artery disease and cardiovascular diseases. Species and to correlate them with Vitamin B12 and folic acid levels in tobacco chewers.

Methods: 50 healthy male tobacco chewers aged 30.6 ± 4.7 years with 3 to 10 years (7.37 ± 2.1) history of tobacco chewing were included in this study. Twenty-five healthy male volunteers, aged 26.5 ± 4.8 years, served as controls. All the participants were from the same community and of similar dietary habits. SOD activity was assayed by method of Paoletti et.al, GR using Goldberg Objective: To evaluate the activities of oxidative Spooner method and vit B12 and folic acid. stress parameters like superoxide dismutase (SOD) and Glutathione Reductase in view of Results: Decreased erythrocyte SOD and GR level possible role of the generation of Reactive Oxygen indicates suppressed antioxidant status. A parallel

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significant decrease in these antioxidant enzymes along with decrease in vit B 12 and folic acid level was observed with increase in duration of tobacco chewing. Conclusion: These factors studied play an important role in oral leukoplakia cancers. The possibility of increase in homocysteine in serum, which elevates the risk of coronary artery disease Abstract No.14.28

and probably cardiovascular accidents, has to be studied further. Moreover, providing adequate quantity of vit B12 and folic acid will prevent cytotoxic effects of chewing tobacco and has to be considered as supplementation in the management of these subjects and while deaddiction programmes.

Role of Pro and Anti-inflammatory Cytokines in the Modulation of alcoholic Liver Disease.

Sukhes Mukherjee* , Subir Kumar Das$, and D.M Vasudevan* *Department of Biochemistry, Amrita School of Medicine, Kochi-682041 $ Department of Biochemistry, Agarthala Govt Medical College, Kunjaban P.O., Agartala-799006 Objective: Hepatic fibrosis is an outcome of chronic liver diseases; and alcoholic liver disease (ALD) is associated with elevated liver hydroxyproline content in experimental animal model. Alcohol also mediates clinical diseases by altering host immune function. normal subjects. While TNF- level increased with severity of ALD, TGF-1 level increased significantly in severe stage of ALD patients compared to moderate and mild stages of ALD. Though serum hydroxyproline content level elevated significantly in ALD patients and Methods: Therefore, we investigated the role of correlated significantly (p<0.01) with every tested pro-inflammatory and anti-inflammatory parameter, but did not change with severity of the cytokines on hydroxyproline content in serum of disease. 32 alcoholic subjects, 40 ALD patients and 45 Conclusion: The results indicate that some of the `normal' subjects. initial cytokines that are produced in chronic ALD Results: Liver function tests were significantly set the stage for direct a profiled cytokine response altered in ALD patients. Interleukin (IL)-4 levels either toward a strong proinflammatory Th1 decreased significantly, while IL-6, IL-10, TNF- cytokine response or a profibrogenic Th2 cytokine and TGF-1 levels increased significantly in response and thus regulate fibrogenesis including ALD patients in comparison to alcoholic and stimulation for hydroxyproline production. Abstract No.14.29

A Simple Urine Test for Diagnosis and Day to Day Management of Diabetes Mellitus

Dr. M.V. Imbichi Mammi Former Prof. of Medicine & Research, Government Medical College, Melevallopra, Kovoor, Calicut, Kerala 673 008. Email: [email protected] Objective: Most of the diagnostic tests for comfortable to the doctor nor friendly to the Diabetes Mellitus available today are neither patient. Our objective is to offer a cost effective

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and hassle-free test to empower patients for single colour varied in proportion to the glucose management, prevention and even cure of the concentration. disease. Conclusion: It is found that the renal threshold Methods: We have experimented and modified for glucose is around 150mg% and not 180mg% the use of GOD/POD solution for estimating the as argued earlier. Painful, costly and invasive blood glucose in urine. Through further trials we have glucose estimation is no more necessary. The simple urine test empowers the patient for succeeded to standardise the new test. diagnosis and day to day management of DM. It is Results: Even 1 mg% of glucose was detected by Cheap, simple, hassle free and is best suited for auto analyser and 10 to 15% of the Benedict's -ve mass screening. cases were shown to be +ve. The intensity of the Abstract No.15.1

Antidiabetic Activity of a Coastal Mangrove, Rhizophora mucronata Poir. : Effect on Carbohydrate Metabolism in Alloxan-induced Diabetic Rats

a

K. Desigamanib, S. Manivannan*, K Kathiresana and T. Ramanathana rita mary arunab Centre of Advanced Study in Marine Biology, Annamalai University, Parangipettai-608502, Tamil Nadu. bAssistant Profesor, Department of Biochemistry, Vinayaka Mission Medical Collage & Hospital, Salem-636308, Tamil Nadu. Email: [email protected]

Results: Oral administration of RBEt (120 mg kg1 ) modulated all the above parameters to normal level. A comparison was made between the actions of RBEt and commercial antidiabetic drug glebenclamide (0.1 mg Kg-1) and the effect of RBEt (120 mg kg-1 ) was near to commercial drug, Methods: Three different concentrations (30, 60, glibenclamide. 120 mg kg -1) for 30 days on blood glucose, haemoglobin (Hb), glycosylated haemoglobin Conclusion: Our study provided evidence for the (GHb), some key carbohydrate metabolic enzymes antidiabetic activity of ethanolic RBEt that can also and liver glycogen in normal and alloxan-induced be extrapolated to clinical studies in future. diabetic rats were studied. Aim: The aim of the present study was to investigate the effect of every day oral administration of ethanolic extract of a coastal mangrove plant Rhizophora mucronata bark extract (RBEt) Abstract No.15.2

Antitumour Activity of Some Homoeopathic Medicines in Animal Models and Possible Mechanism of Action

Ramadasan Kuttan, Preethi K.C., Girija Kuttan Amala Cancer Research Centre, Thrissur, Kerala. Email: [email protected] Homoeopathic medicines are practiced in many Western as well as in Eastern countries. Even though clinical use of homoeopathic medicines against several diseases has been well documented its scientific acceptability is still questionable due to the lack of well carried out clinical trials as well as evaluation of these medicines in a scientific manner. Several homoeopathic medicines have

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been prescribed in cancer. In the present study we have evaluated the use of homoeopathic preparation in the laboratory using modern techniques. Initial studies conducted to find out the cytotoxic potential of potentiated homoeopathic drugs indicated that the some of the potentiated homoeopathic medicines produced cytotoxicity to various cell lines and inhibited T-thymidine uptake. Moreover these preparations were found to inhibit the tumour development induced by Dalton's lymphoma ascites (DLA) cells and Ehrlich ascites carcinoma cells in mice. Potentiated preparation of Hydrastis was found to cure the developed tumours in mice produced by DLA cells. Potentiated homoeo drugs were also found to inhibit the chemical carcinogenesis induced by 3-methylcholanthrene and nitrosodiethylamine. Abstract No.15.3

Mechanism of action of homeopathic drugs indicated that some of the mother tincture as well as their potentiated preparation had similar activity. For example mother tincture of Ruta as well as its 200C potentiated preparation induced chromosomal aberrations and DNA damage in vitro and in vivo. Moreover some of the preparations were found to induce apoptosis to tumour cells as seen in morphology and induction of marker enzymes as well as by Tunnel assay. This was also substantiated by microarray analysis in which several genes involved in apoptotic pathway were found to be elevated by treatment with these preparations. It was also found that some of these preparations induced ROS generation in the cells and this may be a major initiating agent to induce DNA damage as well as apoptosis.

Radioprotective and Chemoprotective Effect of Carotenoid Meso-zeaxanthin A.P. Firdous and R. Kuttan Amala Cancer Research Centre, Amala Nagar, Thrissur, Kerala [email protected] Methods: Radio protective and chemo protective activities of carotenoid meso-zeaxanthin [(3R, 3'S)-â, â-carotene-3, 3'-diol] were studied using in vivo mice models. For radio protective study whole body irradiated (300 rads, single dose) animals and for chemo protective study, cisplatin (16 kg /kg.b.wt, ip, single dose) treated animals were used. Animals were pre treated with different doses of meso-zeaxanthin (50 and 250 mg/ kg.b.wt, orally) and drug treatment was continued after irradiation / cisplatin injection and the animals were sacrificed at different time periods. Results: Treatment with meso-zeaxanthin significantly reduced myelosuppression in both cases as seen from the increased WBC count, bone marrow cellularity as well as number of maturing monocytes in the drug treated animals. The levels of antioxidant enzymes like superoxide dismutase,

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catalase, glutathione peroxidase as well as glutathione levels were found to be drastically decreased after irradiation and cisplatin alone treatment .The levels were markedly elevated by meso-zeaxanthin treatment in a dose dependent manner. Kidney function markers in the serum like urea and creatinine were increased in cisplatin alone treated animals and were decreased significantly by meso-zeaxanthin treatment. The result of histopathological analysis also support the chemo protective potential of meso-zeaxanthin. Conclusion: All these results showed that mesozeaxanthin has profound chemo and radio protective potential. Acknowledgment: This work was supported by a grant from Omni Active Health Technologies Pvt. Ltd., Mumbai.

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Abstract No.15.4

Hepatoprotective Activity of Carotenoid Lutein

Sindhu E R and Ramadasan Kuttan Amala Cancer Research Centre, Amala Nagar, Thrissur, Kerala. [email protected] Aim: Carotenoid lutein (3,3'-dihydroxy- â, å- conjugated diene and hydroperoxides in the liver carotene) was evaluated for its hepatoprotective tissue produced by the administration of activity in rats. paracetamol were found to be reduced in lutein treated groups. The hydroxy proline which is an Methods: Paracetamol (3g/ Kg body weight, indicator of fibrosis in liver tissue was high in orally), 20 % ethanol (7.5 g/kg body weight, control group treated with ethanol. Hydroxy orally) and CCl4 (2.5 ml/kg, ip) were used to proline levels were decreased by simultaneous induce the liver toxicity. lutein administration. The antioxidant enzymes like Results: The level of SGOT, SGPT and ALP SOD, catalase, GPx and glutathione levels in liver which were increased in the serum was found to tissue were found to be increased in lutein treated be significantly reduced by the treatment of lutein groups compared with control group during alcohol 50, 100 and 250 mg/kg b.wt in a dose dependent and CCl4 induced liver toxicity. manner indicating that lutein could reduce hepatotoxicity induced by paracetamol, ethanol and CCl 4. The serum bilirubin was also significantly low in lutein pre-treated groups when compared with control indicating its protective role. Moreover albumin globulin ratio was found to be high in lutein treated groups compared control groups. Increased lipid peroxidation, Abstract No. 15.5 Conclusion: These results substantiate the hepatoprotective action of lutein which may be due to its ability to scavenge reactive oxygen radicals. Acknowledgment: This work was supported by a grant from Omni Active Health Technologies Pvt. Ltd. Mumbai.

XRCC ­ 1 Polymorphisms (Arg194Trp and Arg399Gln) in Oral Cancer from a Kerala Population.

Balaji Natarajan Background : Oral cancer is one of the leading cancers in India. In particular, the state of Kerala has one of the highest incidence of oral cancer in the world. The status of DNA repairing gene XRCC1 genotypes Arg194Trp (exons 5, 6) and Arg399Gln (exons 9, 10) in Oral cancer is to be determined by the following study. samples were selected from the repository for this study. DNA isolated from these samples was amplified using PCR technique. Restriction digestion of the amplified gene was performed using Pvu II & Nci I enzymes respectively to detect polymorphisms in the two XRCC 1 genotypes. The patients were classified into risk groups (cigarettes, betel quid, pan, alcohol or tobacco) and no-risk groups Methods : Tissue samples (normal & tumor) were based on their lifestyle. collected from individuals diagnosed with carcinoma at The Head and Neck Institute. 15 tissue Results: The results so obtained were combined

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with the results available in the lab for the purpose of analysis. So, a total of 30 patients were analyzed. Analysis of exons 5, 6 revealed that only 1 patient was homozygous for Trp194Trp, while 2 (7%) were heterozygous (Arg194Trp) and the rest (90%) showed the Arg194Arg genotype. Exons 9, 10 analysis indicated that 8 (26%) were Arg399Gln heterozygous, while none were homozygous for Gln399Gln. Amongst the risk group, Abstract No.15.6

only 6 patients exhibited polymorphism. Conclusion: Our study showed an increase in the Arg399Arg heterozygote in our study population, suggesting a marginally increased risk of HNSCC associated with this allele, as supported by other studies, especially in other Asian populations. It is of much concern that Arg194Arg was seen in 90% of patients. This is in accordance with other studies that this may be the risk factor.

Multiple Myeloma, 2008 Statistics

Ramesh K, Riju M, Prasanth G, Sreejith A.S., EI Lab Metropolis, North Square, Parmara Road, Opp. To Town Hall, Cochin-18 Email: [email protected] Aim: The study was carried out to determine the incidence of Multiple Myeloma in Kerala population and to study the age and sex wise distribution as well as to analyze the type of immunoglobulin. Methods: Past one year multiple myeloma cases of all sample received for electrophoresis in the EI Lab Metropolis Laboratory was analyzed. Results: Out of the 12000 sample received for electrophoresis 2.5% were having multiple myeloma band. 70% of which were showing thick M Band at gamma region while 15 % were between gamma and beta region and the remaining 15% at beta region. The male female ration was 1.1 suggesting a slight prevalence of the disease in male Abstract No. 15.7 population. Number of case at age below 40 were very less 1% (100% Male), between 41 and 50 were 9% (60% Male, 40% Female), between 51 and 60 were 21% (43% Male, 57% Female), between 61 and 70 were 32% (41% Male, 59% Female), between 71 and 80 were 27% (58% Male, 42% Female) and between 81 and 90 were 10% (78% Male, 22% Female) Conclusion: The incidence of multiple myeloma in Kerala population may be higher than the national and international 0.8 to 1.2 per lakh. Even though there is a higher incidence there is no considerable gender difference in this population. Most of them gives a band in the Gamma region.

Auto Immune Thyroid Disease Antibodies

Riju M, Prasanth G, Sreejith A S. Ramesh K, EI Lab Metropolis, North Square, Parmara Road, Opp. Town Hall, Cochin-18 Email: [email protected] Aim: The objective is to study the pattern of thyroid antibodies in thyroid patients of Kerala. Methods: The seven analytes of the thyroid profile (thyroid stimulating hormone (TSH), Triiodothyronine(T3), Thyroxine(T4), Free T3

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(FT3), Free T4 (FT4), Thyroglobulin antibody(ATA) and anti thyroid peroxidase antibodies (anti-TPO)) were estimated and 218 samples with anti-TPO values above the normal cutoff of 60 IU/ml were analyzed. All samples were analyzed using direct chemiluminescence method on Advia Centaur (Siemens). Results: 91% of the cases were female while 6% male and 3% were children. The incidence is highest during the third decade of life in females and during the 4 decade in men and then shows a decline. 77% of the case were having a Anti TPO values between 60 and 500 and 16% were having very high titre of more than 1300 IU/ml. Almost Abstract No.15.8

50% of the cases were showing a Normal ATA level, 32% had an ATA between 61and 250 IU/ml while 12% showed a high titre of more than 500 IU/ml. 10% of the total were showing a high titre of both Anti TPO and ATA. The average TSH level were 5.95. Conclusion: The incidence of autoimmune thyroid may be high in Kerala population which needs demographic studies. Autoimmune antibodies were predominant in female of reproductive age group, may be mainly due to the postpartum hypothyroidism. There is considerable percentage of high autoimmune titre patients.

Orocecal Transit Time and Small Intestinal Bacterial Overgrowth in Patients with Microscopic Colitis

SV Rana, KK Prasad, SK Sinha, S Sharma, J Kaur and K Singh. Department of Super Specialty, Gastroenterology, PGIMER, Chandigarh Background: Microscopic colitis presents with a characteristic clinical feature of chronic nonbloody watery diarrhea. Small intestinal changes contributing to its pathogenesis and clinical picture have not been extensively studied. There may be delayed orocecal transit time (OCTT) resulting in small intestinal bacterial overgrowth (SIBO) in microscopic colitis. inflammation in the lamina propria with mainly mononuclear cells, a subepithelial collagen layer <10 µm or a diffusely disturbed and thickened subepithelial collagen layer >10 µm, increased number of intraepithelial lymphocytes <20 / 100 surface epithelial cells. Orocecal transit time was measured by non-invasive lactulose hydrogen breath test and SIBO by glucose hydrogen breath test. 100 age and sex matched apparently healthy Aim: Therefore, this study was planned to measubjects were also enrolled. sure OCTT and SIBO in this condition. Results: Mean age of patients was 40.5±15.6 yrs Methods: 54 adult patients with age range 19-75 while of controls 42±17.5 yrs. SIBO was in 7% yrs, complaining for watery diarrhea, were diagpatients while 1% in controls. Orocecal transit time nosed for microscopic colitis by Histopathology. was 112 ± 15.4 minutes in patients and 104.6 ± Out of 54 patients 42 (77%) were males & 12 12.8 minutes in controls. (23%) females. Criteria for diagnosis of microscopic colitis was intraepithelial lymphocytes Conclusion: This study indicates that there was (IEL) >20 / 100 surface epithelial cells, epithelial no significant difference in OCTT & SIBO in padamage such as flattening and mucin depletion, tients with microscopic colitis as compared to controls.

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Indian Journal of Clinical Biochemistry Supplement, 2009/24

Abstract No.15.9

Plasma Myeloperoxidase in Diabetes Mellitus

Usha Anand, Vijaya D, Gayathri B, Aruna V and Anand CV Department of Biochemistry, PSG Institute of Medical Sciences and Research, Coimbatore-641004. Email: [email protected] Objective: Myeloperoxidase (MPO) is an enzyme normally present abundantly in neutrophil granulocytes. It has been shown that MPO serves as a link between inflammation and cardiovascular disease (CVD). It is known that endothelial dysfunction develops early in diabetes mellitus (DM), preceding clinically detectable atherosclerosis. This study was aimed at determining the levels of plasma MPO in patients with an established diagnosis of type 2 DM. Methods: Plasma MPO was determined in 100 patients with type2 DM and 43 controls by a spectrophotometric method. HbA1c and plasma fasting glucose were estimated on Cobas Integra 400 plus autoanalyzer.