Ind J Clin Biochem (Dec 2012) 27(Suppl 1):S1–S114 DOI 10.1007/s12291-013-0298-z

KEYNOTE ADDRESS

RISK MANAGEMENT BEYOND PATIENT SAFETY

Endang Hoyaranda Prodia Group - Jakarta – Indonesia

Managing risk in the medical laboratory is not merely iden- involve e.g. medical record data loss due to insufficiently tifying and controlling laboratory error sources which cause functioning back-up systems, unsustainability of manage- direct harm to patient, e.g. a hyperglycemic glucose result ment functions due to change of officers or malpractice when the patient is hypoglycemic. This aspect of risk to or fraud by officers, etc. patients has been a subject of various quality control and A risk management program which covers the prevention quality assurance as well as certification and accreditation and mitigation of all possible hazards, even indirectly, to programs and education sessions for decades. The medical the healthcare system, should be in place at all medical laboratory professionals worldwide has become increasingly settings including laboratory medicine. aware and knowledgeable about the potentials of direct hazards to the patient by understanding various principles Among various mechanisms to manage risk, the medical and techniques like Pareto, FMEA (Failure Modes and Ef- laboratory management should verify the frequency vs the fects Analysis) and FRACAS (Failure Reporting, Analysis impact of each identified hazard in each type of possible and Corrective Action Systems). risk like the security risk, competency risk, business risk, etc, and then categorize each risk against action taken Apart form the direct hazards imposed to the patient, a if risk arise. The whole process should be implemented number of other risks may indirectly affect the patient. in the routine system to prevent hazards arising without Although indirectly affecting patient safety, these risks being noticed. may be recognized too late to prevent harm. This may

123 S2 Ind J Clin Biochem

AWADHESH SARAN MEMORIAL ORATION

INTEGRATED YOGA FOR THE MODERN LIFESTYLE

Dr HR Nagendra

Abstract: In today’s world health scenario there are many signals are released. Immediately after injury, endorphins factors driving the growth of diseases, but most experts allow animals to feel a sense of power and control over agree that changes in lifestyle including diet are the major themselves that allows them to persist with activity for attributes. As developing countries rapidly industrialize, an extended time people tend to do work involving less physical activity. In Yoga, ‘Cure’ is possible due to the treatment of the root At the same time, the availability of food that is cheap cause of the disease. Through Yoga emotional imbalances but high in calories becomes more common. Stress and are brought down by the art of sublimation of emotions. other psychological phenomena also play a key role in They eradicate the root cause of the diseases, Ādhi. Ad- widespread illnesses. The trend emerging is that the age vanced yogic techniques harness energy to bring about ranges have been dropping and people are getting sick powerful healing of dreaded diseases like Cancer. in the prime of their life. Further, Yoga can create an enhanced feeling of well- Medical professionals base their practice on this matter- being and take us to the state of ever-lasting, permanent based concept using pharmacological and surgical interven- bliss! tions which give symptomatic relief but not the cure as these NCDs have multidimensional etiology. How does Yoga work? According to Taittiriya Upanishad, human beings live a 5-sheathed existence! The Annamaya Yoga offers solutions as it is multi-dimensional; hence can Kosha, Pranamaya Kosha, Manomaya Kosha, Vijnanamaya offer a total solution. It is here that India has to make Kosha and Anandamaya Kosha! The Annamaya Kosha cor- its great contributions, as India has the complete knowl- responds to the body. The Pranamaya Kosha corresponds edge base, which is contained in the Vedas. Coming from to the life-energy which manifests as breath. Manomaya the root vid jnane, the Vedas treasure us with the total kosha corresponds to the mind, Vijnanamaya kosha to knowledge - a knowledge-base much needed for us in the the intellect and Anandamaya kosha to bliss! Yoga helps modern era to meet the challenge. us progress from the grosser physical instincts to the Derived from the verbal root Yuj, the term yoga means divine, subtle bliss. joining; joining our small individual personality with all Swami Vivekananda categorized Yoga in to four streams pervasive cosmic personality; raising us from an animalistic including Raja Yoga, the path of Will Power, Karma Yoga level to the highest levels of perfection, featured by total or the path of Action, Bhakti yoga or the path of Emotions freedom, knowledge and bliss as shown below. and Jnana yoga or the path of Intellect. Based on these Yoga as is commonly understood is not merely practices four streams of yoga, we have developed an Integrated of Asanas as physical exercises. Yoga is a lifestyle where Approach to Yoga Therapy (IAYT) to deal with all Koshas. we are able to live without any conflict, in a state of Our efforts have been to develop specific yoga modules harmony with ourselves and with others around us. Yoga for all NCDs consisting of generalized stress release and leads ultimately total freedom, total power, total knowl- life-style change modules couples with specific modules edge and total bliss! for specific diseases.

All of us know that Endorphins function as neurotransmit- Research Evidence: A brief summary of the research ters. They resemble opiates in their abilities to produce done over the last THREE decades published in indexed, analgesia and a feeling of well-being. We now know that peer-reviewed national and international journals will be when a nerve impulse reaches the spinal cord, endor- presented. phins that prevent nerve cells from releasing more pain

123 Ind J Clin Biochem S3

MRS & PROF. G. P. TALWAR ORATION

DISEASE MODULATING ROLE OF “MEMBRANE COMPLEMENT REGULATORY PROTEINS” AND THEIR SIGNIFICANCE AS EMERGING BIOMARKERS OF SYSTEMIC AUTOIMMUNE DISORDERS

Nibhriti Das Department of Biochemistry, AIIMS

In the late 19th century, Hans Ernst August Buchner Complement mediated tissue injury due to exagger- found that blood serum contained a “factor” or “prin- ated complement activation is a key mechanism in the ciple” capable of killing bacteria. In 1896, Jules Bordet, a pathophysiology of autoimmune disorders like Systemic young Belgian scientist in Paris at the Pasteur Institute, Lupus Erythematosus (SLE) and Rheumatoid arthritis (RA)., demonstrated that this principle had two components: one Animal experiment have, suggested disease modulating that maintained this effect after being heated, and one role of complement regulatory proteins in these disorders. that lost this effect after being heated. The heat-stable Their potential as biomarkers and therapeutic targets had component was responsible for the immunity against also been envisaged. These aspects, however, had not specific microorganisms, whereas the heat-sensitive (heat- adequately been explored in humans. labile) component was responsible for the non-specific In India, triggered by the pioneering work of Professor LM antimicrobial activity conferred by all normal serum. This Srivastava in the Department of Biochemistry, AIIMS, we heat-labile component is what we now call “complement” took up studies to assess the relations of Erythrocyte CR1, earlier known as “alexine.” The term “complement” was soluble CR1 and urinary CR1 with the pathophysiology of introduced by Paul Ehrlich in the late 1890s. glomerulonephritis and their significance as prognostic and Today, complement, in immunology, is a complex system of differential markers of different classes of nephritis,. more than 30 soluble and cell surface proteins comprising We developed a robust method for monitoring soluble of zymogens, receptors and regulatory proteins. Zymogens CR1 levels for our studies and, also a C1q-based Mancini on activation by immune complexes and specific molecular technique to detect and quantitate immune complexes in patterns by antibody dependent classical pathway and serum samples. antibody independent alternative and, MBL pathways generate inflammatory anaphylotoxins & chemotaxins We observed marked declines in the levels of urinary CR1 C3a, C5a, opsonins C3b, C4b and membrane attack com- in patients with SLE which correlated very significantly with plex C56789n, that act in concert to eliminate infectious the glomerular expression of CR1. This observation though microorganisms causing the lysis of foreign and infected unexpected, brought us to suggest and report urinary CR1 cells, the phagocytosis of foreign particles and cell debris, as a marker of glomerular involvement in SLE. and the inflammation of surrounding tissue. Exaggerated With this finding, we, for the first time envisaged disease activation however, bring about cellular damage and acquired impairment of CR1 synthesis in SLE. Earlier, stud- pathological manifestations to the host. A large number of ies remained confined only to E-CR1 and modulation in the Complement regulatory proteins protect the host against levels of cell surface CR1was considered to be due only complement mediated injury in normal health. Interest in to shedding of CR1 from the cell surface by proteolytic membrane-bound complement regulatory proteins geared cleavage. up with the discovery of complement receptor1 (1953, RA Nelson) followed by several others like Decay accelerating Inheritance of low CR1 levels in this disease could be ruled factor (DAF,. Hoffman, 1969.), Membrane Cofactor Protein out by our gene polymorphism studies using PCR-RFLP. (MCP, Cole et al. 1985), CD59 (Protectin, Sugita et al. This led us to focus on Lekocyte CR1 for further stud- 1988;) and few others. ies. Our case-control and longitudinal studies suggested

123 S4 Ind J Clin Biochem

reduced levels of leucocyte cell surface CR1in patients disease. Our observations also suggested that all these with SLE. We found this decrease to be due to reduced membrane complement regulatory proteins hold promise synthesis, impaired translocation of cytoplsmic CR1 to cell as biomarkers of SLE and RA. MCP and CD59 may serve membrane and enhanced proteolytic cleavage of the cell as discriminative markers between SLE and RA. Leukocyte surface CR1. complement regulatory proteins are now known to trigger cell signaling in T- and B-cells Understanding this pleiotro- Simultaneously we found significant correlations of leuko- pic role may bring new insight to the etiopathogenesis of cyte CR1 transcript and protein with SLEDAI scores and autoimmune disorders. other disease related parameters. Translational implications of our findings will fully be rea- Leucocyte CR1 transcript, thus, was envisaged as emerging lised on further studies and a multicentric approach for disease activity marker of SLE. validation of these proteins as biomarkers and establish- Studies were extended to other leukocyte complement ment of reference ranges. regulatory proteins, namely, MCP, DAF and CD59 on the I express my sincere thanks to the organizers and the same lines and also to RA. Aim was to assess a) the inter- award committee who decorated me with this award. relations amongst these proteins and their correlations with the pathophysiology of SLE and RA singly and, in combi- I am deeply grateful to my mentor Professor LM Srivas- nation; b) modulation of these proteins during phases of tava, the “father” of complement research in India ‘for flares and remission and, c) potential of these proteins as initiating, guiding, encouraging and supporting me in this disease activity markers for these two immensely debilitat- fascinating field of research; and, considering me befitting ing prototypypic systemic auto immune disorders. for this award. I have no words to express my gratitude to Professor GP Talwar and Mrs. Talwar. I feel overwhelmed We found increased expression of all these proteins in SLE and honored to receive this award which bears his name during flares excepting CR1 which declined, with significant along with Mrs. Talwar. I express my heartfelt thanks to correlations with SLEDAI scores and other disease activity both of you. On this special day. I gratefully remember parameters and near normalization of the levels during Prof. PS Krishnan, Professor GG Sanwal, Prof. CR Krishna remission. In RA, all these proteins declined with increases Murti and all my teachers for their mentoring and educat- on remission in drug responsive patients. ing me with utmost love and care. I owe this day to my To conclude, expression of leukocyte complement regu- parents who have left the world but are always with me latory proteins CR1, MCP, DAF and CD59 is modulated showering their blessings. I thank all my students, col- along with the modulation of disease activity of SLE and leagues and above all, the volunteers, the, patients and, RA. Their correlations with the disease activity indices the funding agencies, CSIR, ICMR, DBT and DST without were found as significant. This suggested intimate rela- whose dedicated work, co-operation and support our re- tions of these proteins with the pathophysiology of the search work was not possible.

123 Ind J Clin Biochem S5

PATTABHIRAMAN ORATION

PLENARY OF AGEING

Dr. M.V. Kodliwadmath Prof. & HOD, Dept. of Biochemistry, Navodaya Medical College, Raichur, Karnataka

• Ageing is a process of gradual, progressive and gen- Hence this decline can be slowed down or even reversed eralised impairment of functions resulting in the loss at any age by proper understanding of of adaptive response to stress and increasing the risk Bio-physiological changes of ageing, the factors and of diseases. World is now carrying a heavy rucksack theories of ageing, by proper insight to the geriatric of elderly population with its mass increasing every nutrition and with many recent advents in science & moment and by the year 2025 it is expected to rise technology providing a boon to the field of medicine, above 1.2 billion has helped to eliminate risk factors and has provided • Ageing is a normal, but inevitable phenomenon that life prolonging interventions. every human has to undergo. A fact that hurls on us • With a GOAL of disease and disability free life with an ill feeling because of the problems and diseases high physical and cognitive function for the elderly and associated with ageing. Functional capacity of Human placing high importance on exercise and throwing light declines as age advances, but the rate of decline is on its benefits in retarding ageing phenomenon so as accelerated by negative adult life style factors, like to provide healthy, independent and functional life for habits, improper diet as well as environmental and the elderly. external factors.

123 S6 Ind J Clin Biochem

SETH GS ORATION

PHYTOCHEMICALS: A POSSIBLE HERBAL DRUG TO DIABETES AND ITS COMPLICATIONS

Prof. S.B. Sharma Department of Biochemistry, University College of Medical Sciences (Delhi University)

Phytochemicals are non-nutritive plant chemicals that have imparts blood glucose-lowering effect through improved protective or disease preventive properties which includes insulin levels and control over carbohydrate metabolizing mainly antidiabetic, anti-inflammatory, anti-oxidative and enzymes. FIIc also showed protective effect on hypergly- anti-atherosclerotic activities etc. Therefore, the study cemia-induced oxidative stress and endothelial dysfunction was conducted to isolate and characterize the active and hence, it may have beneficial effect in reducing the risk compounds (phytochemicals) from Eugenia jambolana (EJ) of development of cardiovascular diseases. CA100 exerts and Cassia auriculata (CA) effective for antidiabetic and anti-hyperglycemic effect through insulin response along anti-atherosclerotic potential. Detailed UV, NMR, IR spectra with improvement in insulin signaling by reversal of PKC suggested that purified active compound (FIIc) isolated activation and GLUT4 expression and translocation. Both from fruit pulp of EJ is α-hydroxysuccinamic acid (patent FIIc and CA100 treatment cause reversal of fatty steato- granted) and compound isolated from leaves of CA is a sis in heart myocardium. The improved insulin signaling coumarin derivative (CA100). The effective dose of FIIc is pathway and decreased activity of HMG CoA reductase found to be 15 mg/kg b.wt, while of CA100 is 20 mg/kg by CA100 might be responsible for antidiabetic and anti- b.wt., which exhibit glycemic control and lipid-lowering atherosclerotic effect. LD50 of FIIc and CA100 is found to activity is STZ-induced diabetic rats. A single dose of these be much higher than the effective dose, thus they can phyochemicals exhibit glycemic control up to 48 hrs, while be considered quite safe which is also proved both by other conventional drugs require daily administration. FIIc biochemically and histomorphologically.

123 Ind J Clin Biochem S7

INDUSTRY PRESENTATION

LEAN FOR LABS

Prasad Sawant

A New Paradigm: Today laboratories face mounting pres- What Can Make the Most Impact on the Laboratory sure to increase capacity, eliminate waste, overcome labor Operation? shortages and improve quality. To survive and thrive in the Reducing the steps in a process, physical distance traveled current healthcare environment demands diligent focus to complete those steps, and the complexity (or manual on operations improvement and quality. A new paradigm decision making) around those steps will improve the is emerging to help diagnostic laboratories gain a better overall productivity of the laboratory as well as reduce understanding of their productivity. A key component in errors in the final outcome. The technologies or systems this paradigm is the application of new metrics and a a laboratory selects have a great impact on meeting the group of concepts and comparisons that enable labora- needs expressed by their customers, such as the patients, tories to optimize their operations as they move towards physicians and laboratory staff. “Process Excellence”. In a typical laboratory process, there are three phases: the About PEx: Process Excellence (PEx) allows a systematic pre-analytical, the analytical, and the post-analytical. While approach to quality and operational process improvement. many laboratories work on the pre-analytical phase of test- Process Excellence combines the tools and methodolo- ing as their initial improvement project, the analytical and gies of Six Sigma, Lean Thinking and Design Excellence post-analytical phases of testing also need attention. For to drive dramatic improvements in all areas of business example, bringing in barcode scanners to receive specimens performance. decreases hands-on manipulation of samples and improves Three pillars/overarching methodologies are essential to workflow in that part of the testing process, but will not Process Excellence, starting point being Lean: ultimately improve workflow if an analyzer cannot read • Lean Thinking those same barcodes. • Six Sigma PEx/Lean helps laboratory blend process, product and • Design Excellence people to achieve the lab goals now and in the future. These methodologies play a key role in achieving best This empowers labs to maximize the value of equipment practice and empowering the laboratory to achieve maxi- by also streamlining the process around it and enhancing mum productivity. PEx services helps achieve impressive the productivity of staff working in the lab. operational improvements mainly using lean principles. Lean PEx/Lean can make a difference and reshape lab culture, thinking is a methodology that serves to improve workflow helping drive critical business benefits by cutting the un- by helping to eliminate waste, defined as activities that wanted fat (waste) from the system: do not add value relative to customer need. • Lower operating costs There are eight identified wastes in Lean: overproduction, • Improve productivity transportation, motion, inspection, rework/repair, inventory, • Reduce errors and adverse effects waiting and human intelligence. Eliminating waste is not • Overcome staff shortages by improving productivity exactly a novel proposition, and both ISO and Six Sigma • Reduce the flight of talent by empowering and involv- target waste, but the way that Lean removes waste makes ing workforce it possibly the most radical quality improvement manage- • Achieve top performance in patient care experience ment technique. • Engage physicians

123 S8 Ind J Clin Biochem

INDUSTRY PRESENTATION

LEVERAGING TECHNOLOGY TO REDUCE LAB ERRORS: FOCUS ON DRY SLIDE TECHNOLOGY

Prasad Sawant

Knowledge isn’t defined by accurate results alone. Knowl- tal effects for the patient. Inappropriate samples namely edge that truly impacts patient care includes sophisticated hemolysed, icteric and lipemic samples are routine findings information management and interpretation tools to reduce in the lab and have a profound effect on the lab results the probability of human error, improve operational effi- and in turn patient diagnosis and management. MicroSlide ciencies and increase uptime. This is the quality technology technology by virtue of the multi-layered format ensures that frees your lab to focus on improving patient care minimal interference from hemolysis, icterus (bilirubin) and without compromising results. turbidity (lipemia).

The MicroSlide technology or more popularly known as Another large variable which affects lab results is the Dry Slide Technology is one such technology that ensures Quality of water used in the clinical labs. This can affect minimal manual intervention and reduced error potential. results of routine assays like urea, calcium, ALKP, CK etc. About the size of a postage stamp, it contains all the re- The MicroSlide technology completely eliminates the ef- agents for completing the test. It is immediately available fects of water since they function on reagents being in for use. In dry slide technology, multi-layered reagents the DRY format and do not require any water for analysis are applied to a clear polyester support base cut to the and operations. size of a postage stamp. When plasma, serum, urine or Thus proper choice of technology and automation solutions CSF comes into contact with these dry chemistry layers, can play a major role in reducing spurious lab results thus a reaction occurs which is measured using reflectance ensuring better patient care through accurate and reliable spectrophotometry. diagnosis and treatment. Assay interference by endogenous or exogenous sub- stances is an underrated problem with potential detrimen-

123 Ind J Clin Biochem S9

SYMPOSIA I : INVITED LECTURES

A NEW POINTOFCARE DEVICE FOR MEASUREMENT OF NTPROBNP IN EMERGENCY DEPARTMENT

T. Lepoutre1, F. Thys2, F. Verschuren2, D. Gruson1 1Department of Laboratory Medicine, Cliniques Universitaires Saint-Luc, Brussels, Belgium 2Emergency department, Cliniques Universitaires Saint-Luc, Brussels, Belgium

Background and objectives: Amino terminal pro-brain na- intercept of 3.72 and no significant deviation from linearity triuretic peptide (Nt-proBNP) testing supports the triage (P>0.05). Levels of AQT90® Nt-proBNP were also signifi- of patients suspected of heart failure and point of care cantly correlated with BNP (p<0.001). The upper limit of testing (POCT) may allow a faster delivery of results. Our the reference interval of AQT90® Nt-proBNP was 118 ng/l. study was devoted to the evaluation of a new POCT as- Final diagnosis of ED patients were: 24 heart failure [HF], 9 say for Nt-proBNP measurement in patients admitted to coronary artery disease [CAD], 5 pulmonary embolism [EP], emergency department (ED). 3 pulmonary disorder [PD] and 17 non-cardiopulmonary disease [NCP]). In NCP patients, mean Nt-proBNP levels Methods: Nt-proBNP levels were measured with the were significantly lower than in CAD, EP and HF patients AQT90® FLEX analyser (Radiometer Medical Aps, Denmark) but not from PD patients. in 37 healthy volunteers and in 58 ED patients. Results were compared with a central laboratory assay (Elecsys® Conclusion: Nt-proBNP testing with the AQT90® FLEX 2010 analyser, Roche Diagnostics). Circulating levels of showed a good correlation and agreement with the BNP were also determined (DxI® Beckman Coulter, Alere central laboratory assay and allows a fast and reliable reagents) measurement of Nt-proBNP. Therefore, the AQT90® FLEX Nt-proBNP assay may support an efficient of ED patients Results: The Correlation with the central laboratory assay suspected of HF. was excellent (r=0.99, p<0.0001), with a slope of 1.1, an

NEW TRENDS ON POCT

Dr. Bernard GOUGET SFBC-EFLM Representative; IFCC Treasure; Secretary General, International Francophone Federation of Clinical Biology and Laboratory Medicine (FIFBCML)

Diagnostic tests performed outside the central laboratory a core laboratory, point-of-care testing becomes a more or decentralized testing is generally known as point-of-care viable option for many time-sensitive tests. (POC). Biosensors and new technologies for patient-proxi- POCT has the potential to enhance clinical outcome. POCT mal diagnostic products and the new attitudes of patients encompasses a large variety of IVD products ranging from and healthcare providers and systems will transform the moderate sized instrumented diagnostic systems serving organization and the practice of laboratory medicine for larger institutional uses to single-use, disposable tests more efficient medical service and economic perspective. for individual home use. Near patient tests, including Over the years, its use has increased over the past decades those for blood glucose, pregnancy, fertility, coagulation, in response to pressures for cost-containment, faster results cholesterol, drugs, cardiovascular and infectious diseases, and smaller sample volumes. The drive toward centralized drugs of abuse, and various urine components, are in high laboratory operations has catalyzed the value of point-of- demand as physicians and patients realize their potential care testing in hospitals. Now, with testing centralized in

123 S10 Ind J Clin Biochem for promoting improved healthcare in different segments: trend that is expected to continue. The worldwide Point hospital sector, private practice, primary care settings, and of Care Diagnostic testing sector (POCT) is the fastest home monitoring. Existing new technology areas such as growing sector of the worldwide diagnostic industry. protein sequencing, DNA sequencing, and the human proj- The global testing POC market is around USD 14 billion. ect have to be placed into the context of their impact on It will further grow to USD 16.5 billion in 2016 for a rapid diagnosis new developments and forecasts in POCT. compound annual growth rate (CAGR) of 3.7% between In addition, the drive for wellness and the attention sur- 2011 and 2016. The glucosemonitoring segment of the rounding healthcare reforms will focus on rapid diagnosis POC market reached USD 7.5 billion in 2011. The blood and prevention. However, there are many challenges, which chemistry and electrolyte segments of the POC market must be overcome for POCT to be fully utilized. reached USD 2.2 billion in 2010 and will further grow to The value of immediate results is also evident in testing USD 2.8 billion in 2016 for a CAGR of 4.8% between 2011 for chronic diseases where rapid diagnosis can limit the and 2016. The fastest-growing segment, cardiac markers, spread of disease and lead to prompt initiation of therapy is projected to increase at a CAGR of 14.4 percent, rising in the increasing numbers of diagnosed diabetics, people from USD 1 billion in 2011 to USD 2 billion in 2016. The with cardiovascular disease and other chronic conditions, infectious disease and tumor marker segments have the or in people with infectious diseases. For serious diseases next highest projected growth rates, with both expected such as the human immunodeficiency virus (HIV), the to increase at CAGR of more than 10%. ability to rapidly detect the virus in a single doctor visit Globally, the diagnostic business is typified by intense is crucial in initiating therapy as quickly as possible and competition and areas with high growth. The constant ensuring that patients are aware of their test results. In pressure to reduce healthcare spending is shifting health- many countries studies demonstrated that 25%-35% of those care utilization in favor of the medical laboratory, making tested for HIV, using nonrapid methods did not return it an ever more valuable part of the treatment plan. As for test results! hospital stays are shortened, contact between the physician Factors driving growth are the increasing patient and and patient is reduced, which places a larger role on labs physician demands for more efficient testing approaches, to gather, interpret, and deliver accurate information in a availability of new biosensors technologies for more ef- timely manner. POCT is a challenge for our professional fective diagnosis and triage, and reduction of healthcare maturity. Implementing POCT is complex and requires a costs. Although the benefits of POCT are clear, this cooperative effort at many levels. The specialist in lab med industry is not without challenges. Tests performed on has the expertise; we should embrace and manage the clinical laboratory analyzers continue to be perceived as POCT concept. Of course, Universal connectivity of POCT superior to POCT. Companies entering the POCT market devices is the key to managing quality and accreditation, must demonstrate that their tests have more clinical utility according to ISO 22870, which is a part for a successful than those performed in laboratories. POCT implementation. But the challenge to succeed in the task of establishing POCT must be met, so that in POC testing appears to be headed for an even bigger role this 21st century medical laboratory science can exists for in diagnosis and monitoring patient care and the world professionals who practice it. market for POC tests has shown consistent growth, a

123 Ind J Clin Biochem S11

ISCHAEMIC HEART DISEASE AND INSULIN

Udayan Ray Royal Hobart Hospital, Hobart, University of Tasmania Australia

Aim: To study the role of insulin in acute ischemic heart sensitizer. Circulating concentrations of adiponectin are disease (AIHD). determined primarily by genetic factors, nutrition, exercise, and abdominal adiposity. Adiponectin concentrations are Introduction: Acute Ischemic Heart Disease (AIHD) is the lower in subjects with obesity, metabolic syndrome, and main killer disease in the world. With the increase inci- cardiovascular disease. Adiponectin knockout mice manifest dence and prevalence of diabetes mellitus and metabolic glucose intolerance, insulin resistance, and hyperlipidaemia syndrome AIHD has started taking their tides the same and tend to develop malignancies especially when on for ischaemic heart disease has expressed in the similar high-fat diets. Circulating concentrations of adiponectin fashion as it is well known that AIHD is 3-4 folds common are lower in patients with diabetes, cardiovascular disease, in diabetics and so is the case of metabolic syndrome... and several malignancies. The second in the loop in terms of mortality is cancer and again AIHD is 2-3 folds common among cancer sufferers. Methods: We measured HDL, C-Peptide, Insulin, HbA1c%, However the pathological mediator for these diseases re- Nitric Oxide, Resistin and Adiponectin levels in the blood mains obscure. Since lack of insulin or resistance to insulin of AIHD patients and control subjects (healthy sex and plays the key role in diabetics and now it is being known age matched individuals) by Architect CI 8200, Spectro- in cancer, it could be very significant to show the role of photometric method and Immulyte. insulin in acute ischaemic heart disease. Results: HDL, adiponectin and NO levels were significantly The common markers of insulin resistance are hyperin- low (P<0.05), C-Peptide, Insulin and resistin levels were sig- sulinaemia, low HDL, elevated HbA1c%, various cytokines nificantly higher (p<0.05) and not complimentary to plasma and endothelial dysfunction expressed as deranged Nitric glucose levels than those observed in healthy controls Oxide (NO) synthesis. Conclusion: Insulin Resistance is a common phenomenon The adipocytokines are biologically active polypeptides in ACUTE ISCHEMIC HEART DISEASE (AIHD). It could be that are produced either exclusively or substantially by the either a causative factor in the pathogenesis of AIHD or a adipocytes, and act by endocrine, paracrine, and autocrine sequel to it. There is greater need of further research in mechanisms. Out of six adipocytokines (VEGF etc) only this area which could bring a dramatic change or modu- Adiponectin is inhibitory and the rest are promoter to lation in the diagnosis as well as management of Acute angiogenesis. Adiponectin is also an endogenous insulin Ischaemic Heart Disease.

ADRENAL INSUFFICIENCY

V. Parameswaran Diabetes and Endocrinology, Royal Hobart Hospital, Australia

Adrenal deficiency is a relatively uncommon endocrine primary cause -- the adrenal gland being affected, or a disorder with an estimated prevalence of 0.01 to 0.02% secondary cause - due to lack of pituitary drive to stimu- worldwide. It is not gender specific and occurs in both late the adrenal gland. It becomes important, therefore, to males and females and diagnosis is usually based on estab- check the hypothalamic – pituitary - adrenal axis function lishing reduced secretion of Cortisol (and/or Aldosterone) to identify the site of adrenal insufficiency. by the adrenal gland. Reliable diagnostic testing protocols for checking adrenal Failure to produce adequate Cortisol could arise from a reserves of Cortisol include either using stimulation test-

123 S12 Ind J Clin Biochem

ing or, in specific cases depending on the clinical history, combined with more sensitive testing can provide better inhibitory tests are carried out. This allows clinical delinea- outcomes for patients with adrenal insufficiency. tion of the site of deficiency, so that appropriate clinical The testing along with specific clinical cases will be pre- management can be undertaken. Conventional testing sented and discussed.

PROLACTIN SECRETION AND CLINICALLY RELATED DISORDERS

V. Parameswaran Diabetes and Endocrinology, Royal Hobart Hospital, Australia

Prolactin is secreted from the anterior pituitary gland Accurate and timely laboratory diagnosis is critical and by the lactotroph cells. Prolactin release is controlled by sometimes spuriously low or high levels can be reported. releasing and inhibiting factors from the hypothalamus. This might arise from analytical issues which in turn may Prolactin can be raised due to several physiological reasons lead to wrong diagnosis being made resulting in unneces- or in some pathological conditions. Consistently elevated sary clinical investigations being performed that are very levels of prolactin commonly results from a micro or expensive and time consuming. Awareness of these prob- macro adenoma in the pituitary. Whilst they are mostly lems and appropriate remedial action will add to quality benign, early diagnosis and management is important as of reporting. other related disorders like infertility, muscle weakness, A review of prolactin physiology and the disorders result- headaches and sometimes visual problems can ensue. The ing from abnormal secretion will be presented along with incidence of prolactinomas is not high (~ 0.02%) but is the related clinical cases that will help delineate the role of most common pituitary tumour diagnosed in patients with the laboratory in the clinical diagnosis, management and MEN-1, an autosomal dominant condition. prognosis of patients with this condition.

123 Ind J Clin Biochem S13

SYMPOSIA II : CARDIOVASCULAR SYSTEM RELATED DISORDERS

EVALUATION AND MONITORING OF PREANALYTICAL ERRORS IN A HOSPITAL BASED LABORATORY

M. Kankra, S. Bhargava, A Manocha, S. Das, L.M. Srivastava Department of Biochemistry, Sir Ganga Ram Hospital, New Delhi, India

Background: Quality is the heart of management for all patient identification, test order inappropriateness, empty laboratories. For appropriate test results the entire testing tube and inappropriate transportation. process in lab is divided into three phases – preanalytical, Results: On evaluation of preanalytical indicators it was analytical and post analytical. The preanalyitical phase is found that inappropriate sample was the most common an important component of the total laboratory quality. error observed (36.7%) followed by discordance between Errors in this phase are generally underestimated in routine tests requested and test entered into computer system practice. The aim of this study was to design and review (18.3%), insufficient sample (12.1%), inappropriate transporta- components of preanalytical phase from time to time in tion (10.3%) empty tube (9.8%), erroneous request (7.7%), an effort to improve the performance of the laboratory. error in patient identification (2.9%), and test order inap- Method: The study included an evaluation of preanalytical propriateness (2.2%). quality indicator data collected from our lab over a period Conclusion: Sample management directly affects patient of one year. A total of 2, 92,000 samples were received care and outcome and also appropriately timed decision of which 5,923 samples could not be processed as per making of the clinician. Error tracking, scrutiny by evalu- our sample rejection policy. ation and working constantly on preanalytical indicators Quality indicators for preanalytical phase were - inappro- are of utmost importance. With the aim of continuous priate sample (hemolytic, lipemic, clotted etc), insufficient improvement the outcome of these indicators by correc- sample, discordance between tests requested and test tive measures over a period of time will definitely help entered into computer system, erroneous request, error in to improve the quality of laboratory services and patient care.

THE HIGH SENSITIVITY CREACTIVE PROTEIN AND LOW DENSITY LIPOPROTEIN CHOLESTEROL IN RISK PREDICTION OF ISCHEMIC HEART DISEASE

Hariom Sharma, Sunil Patani, Bhavesh Sadariya Department of Biochemistry, Govt. Medical College, Bhavnagar, Gujarat

Abstract: Atherosclerosis is an inflammatory process and group were hs-CRP, LDL-C, HDL-C, Total Cholesterol, Trig- inflammation plays a significant role in the development lyceride, Plasma glucose, Ck-MB. The hs-CRP was measured and progression of coronary artery disease. As C-reactive by immunoturbidimetry method and LDL-C was measured protein is considered a marker for inflammation, leading to by direct method. The hs-CRP was significantly elevated in Ischemic Heart Disease, the study investigated hs-CRP and IHD patients (p<0.01) than the control group. Though LDL-C compared it with LDL-C as a better risk predictor of IHD. was also elevated in IHD patients significantly (p<0.05) This study comprised 100 cases of known Ischemic Heart as compare to the control group but, still 55 patients Disease and 50 apparently healthy subjects as a control with normal LDL-C had IHD but their hs-CRP level was group. They were primarily diagnosed by clinical examina- significantly, high. The specificity and sensitivity of hs-CRP tion and further evaluated by Biochemical investigations. were 1.0 (100%) & 0.97 (97%) and same for LDL-C were Laboratory Investigations done in study group and control 0.66 (66%) & 0.44 (44%) respectively. In the present study

123 S14 Ind J Clin Biochem

we found that the half of the IHD patients with normal guideline for its application as a better & single predictor LDL-C levels had significantly elevated levels of hs-CRP. This of IHD than conventional Lipid Profile. shows that hs-CRP is more sensitive and specific predic- Key Words : hs-CRP, LDL-C tor of IHD than LDL-C and it requires to establish global

GENOMICS OF HUMAN HEART FAILURE: PUTATIVE THERAPEUTIC TARGETS

Hari S. Sharma Institute for Cardiovascular Research, Department of Pathology VUmc University Medical Center, Amsterdam, The Netherlands

Tetralogy of Fallot (TF) is a common form of cyanotic and growth were down regulated where as extra cellular congenital heart disease (CHD) where primary correction at matrix genes were up-regulated. Myocardial expression a young age is the treatment of choice. Main features in of fibrosis markers assessed by video-image analysis and tetralogy of Fallot (TF) include right ventricular hypertrophy corrected for hypertrophy showed significantly increased (RVH) and failure of which underlying molecular mecha- interstitial fibronectin (p<0.01), total collagen (p<0.01) and nisms are not yet understood. We hypothesise that various collagen subtype I (p<0.01) and III (p<0.01) in both TF1 and forms and stages of CHD in children are associated with TF2 as well as peri-vascular fibronectin (TF1; p<0.01, TF2; a number of characteristic novel genes (e.g., cell surface p<0.05) and total collagen (TF1;p<0.01) as compared to receptors, signalling molecules and transcription factors respective controls. TIMPS showed down regulated pattern. etc) and a subset of these genes would predict the etiol- Immunostaining of VEGF in cardiomyocytes was increased ogy and molecular phenotype of cardiovascular defect. We (p<0.01) in patients with TF. Morphometric analysis revealed performed microarray analysis on RVB from 4 patients who enhanced vascular density (p<0.05) with unchanged wall underwent primary correction for the ToF (mean age 0.8 thickness and enlarged myocyte cross sectional areas yr) and compared the expression profile of various genes (p<0.01) with linear correlation (r=0.92, p<0.01) with the age with RVB of age matched controls. Total RNA was isolated, in TF-1 patients. Furthermore, molecular pathway analysis subjected to cDNA synthesis and eventually processed for revealed complex gene interaction in both angiogenesis the preparation of labelled cRNA probes which were used and fibrosis pathways. Quantitative RT-PCR analysis of to hybridise Affymetrix U95A expression chips containing mRNAs revealed enhanced expression pattern in TF-2 as ~12,600 genes in separate experiments. After scanning compared to C2. We conclude that RVH is associated with and computer analysis of the expression data in RVB of enhanced myocardial fibrosis and apoptosis and thereby ToF patients was compared to the baseline expression of diminishing cardiac function in patients with TF. Further- respective genes in control. Data were considered signifi- more, up-regulation of genes encoding VEGF seems key cant when the mean expression changed >2 fold. About event contributing to right ventricular hypertrophy and 200 genes showed up and down regulation pattern by failure associated with stunted angiogenesis and targeting setting a threshold of 2 and above. Data analysis revealed these genes and their products may serve as potential that certain group of Genes involved in cell proliferation therapy for patients with heart failure.

EVALUATION OF NON ENZYMATIC ANTIOXIDANTS AND ELECTROLYTE STATUS IN HYPERTENSIVE PATIENTS, ATTENDING CENTRAL REFERRAL HOSPITAL, GANGTOK

Nimkur S Lepcha1, Mingma L Sherpa1, Bidita Khandelwal2 1Department of Biochemistry, Sikkim Manipal Institute of Medical Sciences, Gangtok, Sikkim 2Department of Medicine, Sikkim Manipal Institute of Medical Sciences, Gangtok, Sikkim

Hypertension, a disease of the urban sedentary is creeping Background: The aim of the study was to evaluate non- into the rural outreaches of the nation which may be due enzymatic antioxidant vitamins status and electrolytes in to rapid urbanization and globalization. hypertensive patients attending Central Referral Hospital,

123 Ind J Clin Biochem S15

Gangtok by estimating the levels of sodium, potassium, Results: Serum sodium was significantly increased in hy- Vitamin C and E. pertensive patients when compared to healthy controls (p<0.05), while no statistically significant difference was Material and methods: The blood samples were obtained observed in serum potassium (p=0.5). Both serum magne- from 20 normotensive subjects who served as control and sium and calcium were significantly decreased in hyperten- 35 hypertensive patients with blood pressure ≥140/90mmHg. sive patients in comparison to normal (p<0.01). The non- The electrolytes along with calcium and magnesium were enzymatic antioxidants of the study Vitamin C and E were assayed in serum with standard kit methods and Vitamin also significantly decreased in hypertensive in comparison C and E were assayed in plasma spectrophotometrically. to control (p<0.01) supporting the fact that imbalance in Student’s t-test was used for statistical analysis of the redox status can be attributed to hypertension. results and a p <0.05 was considered to be statistically significant.

SERUM OMEGA6/OMEGA3 RATIO AND MARKERS FOR CARDIOVASCULAR DISEASE

Ruby Gupta, Ramakrishnan Lakshmy, Ransi Ann Abraham, Jeemon P, Dorairaj Prabhakaran, K Srinath Reddy Center for Chronic Disease Control & South Asia Network for Chronic Disease, New Delhi

A very high omega-6/omega-3 ratio promotes pathogenesis resis. Fatty acids in serum were estimated by Gas chro- of many chronic diseases like cardiovascular disease, cancer, matography. The identified Omega -3 fatty acid included inflammatory and autoimmune diseases whereas reduced were 18:3 (Alpha-linolenic acid), 20:5 (Eicosapentenoic acid) omega-6/omega-3 ratio suppresses the effect. Secondary & 22:6 (Docosahexenoic acid). Among omega-6 included prevention studies have demonstrated a decline in disease were 18:2 (linoleic acid), 18:3 (gamma-linolenic acid) & 20:4 progression or reversal of symptoms with decline in ratio; (Arachidonic acid). however the optimal level varies with the disease under Complete data was available for 176 participants (89% males consideration. We have evaluated the circulating levels of and 11% females) with mean age of 47.23 +/- 6.00 years. omega-6/omega-3 ratio and its effect on cardiovascular The bmi of the participants was 24.88 +/- 3.43 Kg/m2 and risk markers in India. waist circumference was 91.50 +/- 9.56 cm. The median of The present study was conducted in industrial setting omega-6/omega-3 ratio in the study population was 36.69 where employees were randomly selected. Data on their (range: 6.21 - 183.69). The levels of total cholesterol, trig- demographic characteristics were collected using pre-tested lycerides, ldl-cholesterol and cholesterol/hdl ratio and apo questionnaire. Fasting blood samples were collected from B/A ratio correlated significantly (p<0.05) with omega6/3 all the participants. Serum/ plasma was separated and ratio. No significant correlation was observed with hsCRP, stored at -800C till the time of analysis. fibrinogen and LDL- particle size.

Lipids were estimated using standard kits. LDL-particle size A direct relationship of omega-6/omega-3 ratio with dys- was estimated using polyacrylamide slab gel electropho- lipidemia was observed in our study.

CARDIAC RISK PROFILE IN AT RISK ADOLESCENTS

Satinder Kaur, Navjot Kaur, GS Wander*, Parveen C Sobti** and M Minocha** Department of Biochemistry, Hero DMC Heart Institute* and Department of Pediatrics** Dayanand Medical College and Hospital, Ludhiana

Objective: Cardiovascular disease although diagnosed in provide significant knowledge about hereditary origin of adulthood, its roots begin in childhood, so the intervention disease. strategy to reduce risk should begin as early as possible. Materials and Methods: 100 adolescents with family his- Risk profile in adolescent children of CAD parents may tory of CAD were enrolled for the study. 100 adolescents

123 S16 Ind J Clin Biochem

without family history of CAD served as controls. Cardiac Conclusion: Although environmental risk factors (smoking, risk factors: smoking, alcohol consumption, physical activ- alcohol consumption, physical activity, abnormal eating ity, obesity, truncal obesity, hypertension, dyslipidemia and habits) were similar in adolescents of CAD patients as dysglycemia were evaluated. of normals, still the metabolic abnormalities like obesity, hypertension and dyslipidemia were significantly higher Results: Smoking, alcohol consumption, physical activity and amongst off-springs of CAD patients thereby indicating a eating non conventional food habits were similar in both genetic predisposition towards abnormal risk factor. Con- the groups. Cholesterol, Random Blood Sugar, Body Mass sequently they need more aggressive prevention measures Index, Waist Hip Ratio and blood pressure was significantly as compared to controls. higher in study group compared to control group. Incidence of obesity (13 vs 8), overweight (9 vs 2), central obesity Keywords: Adolescents, body mass index (BMI), waist (15 vs 6), hypertension (12 vs 5) and hypercholesterolemia hip ratio (WHR), coronary artery disease (CAD), Cardiac (20 vs 10) was higher in study group. risk factors.

SHOULD NON HDLC REPLACE LDLC AS THERAPEUTIC TARGET TO DECREASE CARDIOVASCULAR RISK

Seema Gupta, Jasbinder Kaur, Shivani Jaswal, Harjeet Kaur, Harpreet Kaur Department of Biochemistry, Govt. Medical College and Hospital, Chandigarh

Aims and Objectives: LDL cholesterol is considered as major Results: Waist circumference correlated both with LDL-C risk factor for cardiovascular disease and is the primary and non HDL-C (r=0.31 and 0.41 respectively, p<0.001) but target of lipid lowering therapy. The estimation of LDL-C the degree of correlation was more with non HDL-C. The by Friedwald’s formula becomes progressively less accurate coefficient of correlation between FBS and LDL-C was 0.08 with increasing triglycerides, is not valid for type III hyper- which was non significant (p>0.05) whereas between FBS lipoproteinemia and requires fasting samples. Nowadays, and non HDL-C it was 0.15 and significant (p<0.05). HDL-C non HDL-C is gaining significance as cardiovascular disease showed negative correlation with waist circumference (r= risk score predictor. It accounts for LDL-C, VLDL-C, IDL-C, -0.18, p<0.05) but no significant correlation was found chylomicron remnants and lipoprotein (a). The objective of between HDL-C and FBS (p>0.05) the study is to calculate non HDL-C in sedentary population Conclusions: Non HDL-C had better correlation with waist and find its correlation with FBS and waist circumference circumference and FBS as compared to LDL-C and HDL-C. which are the key components of metabolic syndrome and Serum non HDL-C can be considered as a better screen- predisposes to cardiovascular disease. ing tool than LDL-C for the assessment of CAD risk in Material and Methods: The study group comprised of adults as it includes all classes of atherogenic lipoproteins. 197 adult individuals having sedentary life style. FBS, total Moreover, its value does not depend on triglycerides levels cholesterol (TC), triglycerides and HDL-C were measured and can be used as therapeutic target for patients having by automated Roche assays and LDL-C was calculated by hypertriglyceridemia also. Freidwald’s formula. Non-HDL-C was calculated by equation TC- (HDL-C). The waist measurement was taken above the iliac crest at the natural waistline.

123 Ind J Clin Biochem S17

EVALUATION OF CYSTATIN C AS AN EARLY MARKER OF RENAL IMPAIRMENT IN DIABETIC AND HYPERLIPIDEMIC PATIENTS

Dr. Shyamali Pal Consultant Biochemist & Technical Director, S. Serum Analysis Centre, Serum Group, Bidhan Sarani, Kolkata

Objective of the study: The NKDEP*, NIDDK*, NKF* and of lipid profile and Cystatin C at 4 month and 8 month ASN* recommend estimating GFR from serum Creatinine. after starting 10mg atorvastatin per day was done. The The commonly used equations are MDRD study equation, diet is prescribed by the same dietician, suggested medi- Cockcroft-Gault and CKD-EPI equation. The 4th choice is cine was same (Storvas, 10mg/day), age group was same, cystatin C-GFR equation based on estimation of cystatin having similar lifestyle and no history of renal impairment, C from blood sample and measuring the renal impairment inflammatory diseases. The patients have not used any status from the scale provided in the diagnostic kit. other medicine except Storvas, 10mg after dinner. So, the follow up study is of help to find out the interrelationship The role of cystatin C, the protein encoded by CST3 gene of lipid profile with Cystatin C. as an early marker of renal impairment is established, pro- vided the patient is euthyroid and not on glucocorticoid So, The study has been done in four parts- i) Validity of treatment. Elevated concentration of cystatin C have also cystatin-C-GFR compared to classical creatinine clearance been reported in inducible ischemia and in cardiovascular and other calculated values. ii) Evaluation of Cystatin C as heart disease (CHD) patients not on β-blocker or statin. early marker of renal impairment in diabetics with normal serum creatinine and microalbumin creatinine ratio in the But in Indian scenario where the comparison is within range of microalbuminuria. iii) To assess whether Cystatin C two races seems not to be satisfactory. So, in Group is acting as non renal marker in euglycemic –hyperlipidemic I a comparative study of classical creatinine clearance patients. iv) Pharamacokinetic study on Storvas users. with calculated clearances and cystatin C-GFR has been performed. ABB: NKDEP-National Kidney Disease Education Programme, USA. NIDDK-National Inst. of Diabetes, Digestive and Kid- In studies on patients with CVD without statin cystatin ney Disease, USA. NKF- National Kidney Foundation, USA. c was found out to be elevated and some non renal ASN- American Society of Nephrology, USA. mechanism have been proposed. The association of Cys- tatin C was thought to be either direct pathologic effect Materials and methods: Study materials: 30 Diabetic nor- or inflammatory effect. molipidemic (male), 30 nondiabetic hyperlipidemic (male), of the age group 45-55 yrs were included in the study. 10 The chronic diabetics are having high risk of renal impair- patients of the same age group and sex were included in ment. So, a study was (group II) designed to compare phase three study. The history of diabetes for group II was Cystatin C values with microalbumin creatinine ratio, and 6-8 yrs and they were under oral antidiabetic medicine. cystatinC-GFR values. The statistical analysis of these data and comparative analysis of duration of diabetes, age, glu- The patients in group (III) study were euglycemic. When cose (F&PP) may help to get an idea about the effectivity hyperlipide-mia was detected they were not on any medi- of Cystatin C as early marker of renal impairment. cine. After starting of 10mg Storvas/day the Cystatin C, CRP, creatinine and lipid profile have been estimated at The cystatin C levels of euglycemic hyperlipidemic (group 4 and 8 months, 12months. III) were studied and C-reactive protein (CRP) of the 2nd group have also been estimated. As the cause is not Methods: Tests performed are serum/urine creatinine, Cys- renal so raised cystatin C has to be due to some non tatin C, plasma glucose fasting and post prandial, microal- renal cause. CRP has been estimated to rule out inflam- bumincreatinine ratio (M/Cr) for phase II study. In phase matory cause. III instead M/Cr, CRP has been estimated. In phase III only glucose fasting, CystatinC, lipid profile and creatinine were In group IV, taking 10 euglycemic, hyperlipidemic patients a performed. GFR has been calculated from eGFR/MDRD pilot study was performed. The basal level and estimation

123 S18 Ind J Clin Biochem

calculator and Cystatin C-GFR calculated from the scale of Cystatin C is apparently related to hyperlipidemia. The provided in the kit. Rourine parameters like glucose, lipid results facilitated the study involving group IV. The group profile, serum/urine creatine, CRP etc, estimated in fullybau- IV study, though it is a pilot study has nicely shown the tomated system Cobas-6000 integrated analyser. Cystatin lowering of serum creatinine and Cystatin C after getting C estimated in Beckman CX-5Pro automated system. treatment with 10mg of atorvastatin/day. The decrease in creatinine concentration, improvement of lipid profile Conclusion: In group I&II study it seems comparison of clas- and lowering of CystatinC have been found out to be sical creatinine clearance and Cystatin C-GFR is correspond- directly proportional and statistically the changes are very ing well than calculated values. In group III, the raised level significant. Hence, Cystatin C may be referred as study of of CystatinC is neither associated with diabetes nor any impairment marker in hyperlipidemics also. inflammatory process as the patient group is euglycemic and CRP of the whole group is normal. So, the raised level

EVALUATION OF HIGH SENSITIVE CREACTIVE PROTEIN HSCRP AND SERUM LIPID PROFILE IN HYPERTENSION

Supriya Dawri, R.R. Melinkeri Department of Biochemistry, Bharati Vidyapeeth medical college, Pune

Background: Hypertension is a common, asymptomatic, cine OPD at a tertiary care hospital. A healthy group of readily detectable disease that leads to lethal complications normotensive volunteers were taken as controls. Fasting if left untreated. Vascular inflammation may be involved in blood samples were collected for measurement of serum both the initiation and development of hypertension that is lipid profile and hsCRP (by CLIA). evident from the elevated levels of inflammatory markers Results: There was statistically significant rise in hsCRP like Tumor necrosis factor-α, Interleukin-6 and C-reactive levels in hypertensives as compared to controls (p<0.001). protein (CRP) found in people with hypertension with no The concentration of cholesterol, triglycerides and LDL-C evidence of cardiovascular disease. The ‘high sensitivity CRP were significantly high in cases as compared to controls (hsCRP)’ simply refers to the lower detection limit of the (p<0.001). There was statistically significant decrease in assay procedure. hsCRP is associated with an increased risk HDL levels in cases as compared to control. of incident hypertension at all baseline blood pressures and among individuals without traditional coronary heart Conclusion: Findings of higher levels of hsCRP in hyper- disease risk factors. tension along with atherogenic lipid profile suggests that elevated hsCRP and hypertension can be independent Objectives: The present cross-sectional study is an attempt determinants of cardiovascular risk. to evaluate the relationship of serum hsCRP levels and Serum Lipid profile in hypertension. Keywords: Hypertension, hsCRP, Lipid profile.

Material & methods: The study group included twenty-one diagnosed cases of hypertension patients attending medi-

123 Ind J Clin Biochem S19

SYMPOSIA III : FREE RADICALS AND ANTIOXIDANT

EFFECT OF NBUVB PHOTOTHERAPY AND ANTIOXIDANT SUPPLEMENTATION ON OXIDATIVE STRESS STATUS IN VITILIGO

Brijesh Rathore1, Aparna Misra1, Kshitij Garg2, Devesh Misra2 and Farzana Mahdi1 1Department of Biochemistry, 2Department of Dermatology, Venereology & Leprosy, Era’s Lucknow Medical College, Sarfarazganj, Hardoi Road, Lucknow

Oxidative stress plays an important role in the pathogenesis compared to baseline. While in stable vitiligo patients of vitiligo. Free radicals destroy melanocytes and develop (SVP) additional 4 weeks of combined phototherapy and an aberrant immune response. The altered antioxidant en- antioxidant supplementation resulted in significant (p<0.05) zymes in the vitiligo epidermis, have been reported to be increase in activity by 5.98% compared to its activity at dysregulated. Narrow Band UVB phototherapy (NB-UVB) is 4 weeks. Superoxide dismutase (SOD) activity showed the preferred modality for the treatment of vitiligo. The significant (p<0.05) increase in both AVP and SVP groups role of oral antioxidant supplementation has been sug- at 8 weeks as compared to its activity at 4 weeks as gested to overcome the deleterious effects of free radicals. compared to NB-UVB phototherapy group. The change in The aim of the study was to assess the oxidative stress glutathione peroxidase (GPx) activity from baseline to 4 and antioxidant status in vitiligo patients, after four weeks weeks of NB-UVB phototherapy was 2.10% in AVP and 4.89% of NB-UVB phototherapy alone; and combined NB-UVB in SVP. After 4 weeks of NB-UVB phototherapy there was phototherapy and antioxidant supplementation. an increase in the malondialdehyde (MDA) levels from their baseline values in both AVP and SVP. Thus, it is clear that In active vitiligo patients (AVP), the change in catalase antioxidant supplementation with NB-UVB phototherapy (CAT) activity was 4.56% in NB-UVB phototherapy group may be given to vitiligo patients. and 10.44% in the antioxidant supplemented group as

PROOXIDANT ANTIOXIDANT IMBALANCE IN MENOPAUSAL WOMEN OF NORTH WESTERN INDIAN PUNJABI POPULATION

Jyot Amrita1, Mridula Mahajan2, Sujata Sharma3, Sarabjit Singh4 Department of Biochemistry1, Department of Medicine4, Sri Guru Ram Das Institute of Medical Sciences and Research, Amritsar. Department of Biochemistry2, Department of Gynaecology3, Government Medical College, Amritsar

The state of menopause is marked by many metabolic coronary artery disease. The patient group comprised of changes such as redox-status imbalance, changes in body menopausal women (n=100) suffering from CAD and the fat distribution, dyslipidemia, reduced glucose tolerance, control group included those menopausal women (n=70) hypertension, endothelial dysfunction and vascular inflam- who were not having any evidence of coronary artery mation. Women in the menopausal stage are considered disease. All the females were screened for serum lipid to be at relatively higher risk of developing coronary profile, serum malondialdehyde (MDA), serum superoxide artery disease (CAD). There is substantial evidence which dismutase (SOD) activity and LDL protein oxidation. LDL indicates that oxidative stress plays a key role in the carbonyl content was taken as an index of LDL protein pathogenesis of coronary artery disease and it may be oxidation. It was observed that 42% of the menopausal possible that the prevalence of high oxidative stress in females suffering from CAD were normolipidemics whereas females at initial stages make them more susceptible to 56% of menopausal females in the control group (without the development of CAD at menopause. Hence, the pres- CAD) were hyperlipidemics. This clearly shows that lipid ent study was aimed to evaluate the role of oxidative levels are not always predictive of increased risk of CAD in stress in menopausal women with and without having menopausal women. Therefore, there may be some other

123 S20 Ind J Clin Biochem

hidden detrimental factors involved in the disease patho- may be said that increased oxidative stress at the initial genesis. One of the most important culprits behind this stages (before menopause) may be responsible for making is the oxidative stress. Serum MDA levels were observed some females more susceptible to CAD development at to be significantly raised (p<0.05) in female patients while menopause than others. Hence, it is highly important to serum SOD activity was significantly low (p<0.05) showing keep a check at pro-oxidant-antioxidant imbalance at earlier their disturbed antioxidant status. This was further sup- stages so as to avoid the complications at an unavoidable ported from the observation that LDL carbonyl content stage of female’s life i.e. menopause. was significantly elevated (p<0.05) in CAD menopausal Keywords: Menopause, Oxidative Stress, Coronary Artery women as compared to the control group. Oxidation of Disease LDL initiates the process of atherosclerosis thereby promot- ing increased oxidative stress and hence inflammation. It

ESTIMATION OF TOTAL ANTIOXIDANTS IN VERY LOW BIRTH WEIGHT NEONATES BEFORE AND AFTER RECEIVING TPN

Mithun Nath*, Sumantha H.D*, Anjali Rao*, Kishnananda Prabhu*, Jayashree Purkayastha#, Soundaram V# *Department of Biochemistry & #Department of Pediatrics, Kasturba Medical College, Manipal

Introduction: Very-low-birth-weight infants are born at estimating the total anti-oxidants in neonates before and a time of otherwise rapid intrauterine brain & body after receiving TPN. growth. These infants have limited endogenous energy Materials and Methods:Hemodynamicallystable, low birth stores. Specificneonatal morbidity (ventilator & oxygen weight infants with a birth weight <1250 g are included in dependent infants) may also increase metabolic demands the study and total anti-oxidants in the plasma is measured by as much as 25% compared to the controls. Rapid es- using FRAP (Ferric Reducing Anti-oxidant Property) assay, tablishment of postnatal nutrition is essential to provide on the day of birth as well as at the end of TPN. FRAP continuous administration ofnutrition. Thus total parental assay offers a putative index of antioxidant or reducing nutrition is provided to achieve a body composition & potential of biological fluids. growth similar to those of a normal fetus of the same postmenstrual age. Result: No significant changes in the anti-oxidants level is observed so far. More significant results are expected in Objective:Total Parental Nutrition (TPN) is a potential future as the study is ongoing. source of oxidants. A balance between oxidants & anti- oxidants is instrumental to maintain a competent immune Conclusion: The oxidative stress in the VLBW infants is not system. This study is carried out with an objective of significantly altered before and after receiving TPN.

ACUTE RENAL FAILURE IN LIVER TRANSPLANTS: INDIAN STUDY

Pradeep Naik*, Prem Sagar B, Mallikarjuna M. Department of Clinical Biochemistry, Global Hospitals, Hyderabad

Acute renal failure (ARF) is a frequent medical compli- following indications: biliary atresia (n=1), Wilson’s disease cation after liver transplantation (LTx). We analyzed 68 (n=2), hepatitis B–related liver cirrhosis (n =9), hepatitis (males 57, Females 11) liver transplant recipient’s From C–related liver cirrhosis (n=15), Acute liver disease (n =17), January 2003 to December 2009, a total of 68 patients Chronic liver disease (n=7), Alagille syndrome (n=1), cho- underwent (cadaveric50, live 18) Liver Transplantation. They langitis (n=3), autoimmune cirrhosis (n=2), Non alcoholic had developed ARF early in the postoperative course. Their fatty liver disease (n=10). mean age was 42 years at LTx. The 68 recipients had the

123 Ind J Clin Biochem S21

All patients had received steroids and Mycophenolate, with at 3-4 weeks after the transplant and the mean of serum combination of Tacrolimus (FK506) after liver transplant. MDRD was 98 ml/min. We analyze the MDRD (Modification of Diet in Renal The ARF etiology was multifactorial for the majority of pa- Disease) before and after liver transplant in 68 patients. tients. These factors include arterial hypertension, diabetes The mean of serum MDRD was 95.9 ml/min in before mellitus, hepatitis and immunosuppression. We conclude transplant. All patients developed ARF at a mean time that Immunosupression immediately after transplantation of 5 days after LTx and the mean of serum MDRD was influences the development of acute renal failure. ARF 52.8 ml/min. During the initial days of after transplanta- treatment included fluid replacement, decreased or altered tion renal function deteriorates rapidly by nephro toxic immunosuppressive agents, avoiding exposure to nephro- effects of immunosuppressive drugs. Immunosuppression toxic drugs, and adjusting antibiotic dosages. immediately after transplantation influences the develop- ment of acute renal failure but declines slowly thereafter. Keywords: Acute renal failure, LTx, MDRD. The majority of patients returned to normal renal function

OXIDANT AND ANTIOXIDANT STATUS IN CRF

Dr. Ratna Priya1, Dr. J.R. Keshri2, Dr. Uday Kumar3, Dr. Anand Sharan4, Dr. Rekha Kumari5, Dr. Chandan Kishore6, Dr. Jyoti Priya7 1Senior resident, 2Asst. prof. 3Prof. & head 4Addl. Prof. 5Asst. Prof. 6Asst prof. 7Senior Resident, Dept. of biochemistry, Indira Gandhi institute of medical sciences, Patna. 6Dept of Radiology, Narayan Medical College & Hospital, Sasaram. 7Dept. of Physiology, Indira Gandhi institute of medical sciences, Patna Abstract: Oxidative stress plays a significant role in patho- The level of MDA, creatinine, blood urea nitrogen was genesis of chronic renal failure (CRF). Redox imbalance significantly increased, antioxidant vitamin levels were caused by increased ROS production and/or reduced an- decreased in CRF patients as compared to normal sub- tioxidant reserve, leading to pathological consequences jects. The elevated MDA level can lead to lipid peroxida- including damage to proteins, lipids and DNA. The present tion and can also promote the complication of CRF. The paper focuses on the extent of free radical damage on antioxidant levels were decreased, so these patients can lipid (measured as malondialdehyde, MDA) and effect on be supplemented with antioxidant vitamins for preventing antioxidant defense mechanism (measured as vitamin C) lipid peroxidation and occurance of complication. in the serum of CRF patients predialytically.40 cases and Keywords: Antioxidant vitamins, chronic renal failure, MDA, 40 controls between age group of 25 – 60 were taken. oxidative stress

ASSOCIATION OF ANTIOXIDANT POTENTIAL AOP OF LDL WITH CORONARY HEART DISEASE IN TYPE 2 DIABETIC PATIENTS

Sanjeev kumar singh, Nivedita Singh, Neelima Singh, Harshvardhan Singh Bacground and Introduction: Cardiovascular disease is one Materials & Methods: Serum sample was tested from 90 of the major causes of death in type 2 diabetic patients. Type 2 DM patients in whom 45 were without any compli- The peroxidation of lipoproteins, especially low density cation and 45 with CHD and 50 healthy controls matched lipoproteins (LDL) plays a significant role in the pathogen- for age, height, and weight. LDL from the serum sample esis and progression of atherosclerosis in diabetes. This was precipitated by heparin-citrate precipitation method. peroxidation reaction is countered by antioxidants present AOP of LDL was measured in isolated LDL. in LDL known as its antioxidant potential (AOP). Results & Conclusion: AOP was significantly decreased in Aim of the study: The present study was carried out to all diabetic groups compared to control group. Individuals see the levels of AOP of LDL in T2DM patients with and with type 2 diabetes experience increased risk of athero- without CHD. sclerosis could be due to decreased AOP of LDL.

123 S22 Ind J Clin Biochem

THE EFFECTS OF OXIDATIVE STRESS AND VITAMIN C LEVELS IN COPD AND ASTHMA

Shweta Paul1, Anju Sharma1,3, Kapili Nath1, Indresh Kumar Singh1, Prema Adhikari1, P. Pavni1, Surinder Kumar Bansal2, Ashok Shah4, Vishwajeet Rohil1 1Department of Clinical Biochemistry, 2Department of Biochemistry, 4Department of Pulmonary Medicine, Vallabhbhai Patel Chest, Delhi. 3Department of Biochemistry, Vardhman Mahavir Medical College & Safdarjung Hospital.

Background: The pathogenesis of COPD and asthma has • To measure oxidant (MDA level) and antioxidant (Vit. been claimed to be attributable to increased systemic and C level) in Asthma, COPD patients and controls. local oxidative stress. Asthma and COPD are diseases with • To compare MDA and Vitamin C levels in patients of different physiological and anatomical features. One of Asthma and COPD with control. the hypothesis in the pathogenesis of Asthma and COPD Methodology: A total number of 60 subjects comprising is that the increased oxidant burden due to release of 20 controls, 20 COPD and 20 Asthma cases were studied. reactive oxygen species from airspace leukocytes may not In all the subjects, Plasma levels of MDA (Ohkawa et al., be adequately counter balanced by the lung antioxidant 1979) and Vitamin C (Jagota & Dani, 1982) were estimated systems, resulting in oxidative stress. Markers of the spectrophotometrically. elevated oxidant burden have been detected in both of these disorders. ROS are produced endogenously by several Result and Conclusion: The MDA levels were significantly cell types including activated inflammatory and structural increased in both COPD patients as well as Asthma cases cells, the most potent generators of these species being as compared to controls. In COPD patients as compared eosinophils and neutrophils, i.e., cells that are abundant to controls, the levels of plasma Vit-C was found to be both in Asthma and COPD and we intended to study MDA significantly decreased and in Asthma patients though as a biomarker of lipid peroxidation and antioxidant Vitamin Vit. C levels were decreased but it was not statistically C in plasma of Asthma and COPD cases. significant.

Objectives: Present study is undertaken to evaluate Plasma Keywords: Chronic Obstructive Pulmonary Disease – COPD, levels of Vitamin C and MDA in Controls, Asthmatics and Malondialdehyde – MDA, Reactive oxygen species - ROS. COPD cases.

AUGMENTATIVE EFFECT OF CATECHINS ON INHERENT ANTIOXIDANT CAPACITY : A COMPARATIVE STUDY

Renu Uniyal**, Vinita Kalra* *Professor, Department of Biochemistry, Himalayan Institute of Medical Sciences, Dehradun, India *Reader, Department of Biochemistry, UDMRI, Dehradun, India

Excessive generation of reactive oxygen species (ROS) Results of the study include a significant reduction in the induces oxidative damage to vital cellular molecules and concentration of MDA after treatment with catechins from structures including DNA, lipids, proteins and membranes. the two plant species. Acacia Catechu was more effective Catechin is a flavon-3-ol, a type of natural polyphenol (p ≤ 0.00) in this regard. It was also observed that the belonging the family of flavonoids. The catechins and purified catechin extract caused greater reduction of MDA can be isolated from a variety of indigenous medicinal (p ≤ 0.02 vs. Kattha). This outcome was further correlated plant species. with pronounced augmentation of the inherent antioxida- tive capacity as manifested by increase in the activities This study was aimed to compare the antioxidative po- of the natural antioxidant defense enzymes viz. Catalase, tential in vivo of Kattha & purified catechins derived Glutathione peroxidase and Glutathione reductase. from two different plant species, namely Acacia Catechu & Uncaria Gambier.

123 Ind J Clin Biochem S23

SYMPOSIA IV : INVITED LECTURES

MYCOBACTERIAL SECRETORY PROTEOME WITH ES31, ES41, ES43, ES20, ES38 AND ES6 PROTEINS OF INTEREST AS ANTIGEN, BIOMARKER AND ENZYME WITH DRUG TARGET POTENTIAL

B.C. Harinath JB Tropical Disease Research Centre, Mahatma Gandhi Institute of Medical Sciences, Sevagram,

Tuberculosis (TB) remains a major threat to public health losis sera of patients attending a tertiary care Kasturba worldwide and has been declared as global emergency by Hospital. The antigen assay was found to be more sensi- WHO in 1993. According to the World Health Organization tive than antibody based assay for detecting tuberculosis (WHO, 2009), 9 million new TB cases including 4 million with HIV co-infection. Biochemical characterization showed smear positive cases with 15% HIV- Positivity have been that in vitro released ES-31, ES-43 and ES-20 antigens as reported in 2007. glycoproteins while ES-41 and ES-38 antigens were found to be lipoprotein in nature. The secretory proteome of mycobacterium tuberculosis consists of large number of actively secreted proteins and User friendly Peroxidase ELISA has been developed for enzymes of interest in diagnosis, immunoprophylaxis and detection of circulating SEVA TB ES-31 antigen – a serine as drug targets. In our laboratory proteins SEVA TB ES- protease (Free antigen and Immune complexed antigen) 31, ES-43, ES-41, ES-20 and ES-6 were isolated from short with 70-75% sensitivity and 90% specificity and with a term M. tuberculosis H37Ra culture filtrate by FPLC, affinity sensitivity of detection of antigen is 1 ng/2μl (0.5 μg / ml chromatography and SDS-PAGE and explored in diagnosis. serum). SEVA TB ELISA using cocktail antigen and cocktail The seroreactivity of these purified antigens were assessed antibody is being routinely done for screening of patients in pulmonary tuberculosis, extra pulmonary tuberculosis, suspected of tuberculosis attending Kasturba Hospital, a different stages of pulmonary tuberculosis (fresh, relapse, tertiary health care centre located in rural area. chronic and latent) and in HIV-TB co-infection. Analysis of Further study revealed that ES-31 antigen is a zinc con- immune response to these purified antigens by indirect taining serine protease with protease and lipase activities. and sandwich ELISA using highly sensitive Penicillinase Serine protease inhibitors such as pefabloc, 3, 4 dichlor- enzyme immuno assay, showed ES-31 antigen as having oisocoumarin, phenyl methyl sulphonyl fluoride (PMSF) good diagnostic potential in pulmonary tuberculosis and in and metalloprotease inhibitors such as ethylene diamine certain groups of extra pulmonary tuberculosis in particular tetracetic acid (EDTA) and 1, 10 phenanthroline and orlistat tuberculous lymphadenopathy and tuberculous meningitis, (lipase inhibitor) inhibited 65 – 92 % serine protease activity whereas ES-41 was found to be more seroreactive in in vitro. Isoniazid showed 95 % inhibition on mycobacte- abdominal and bone & joint tuberculosis. ES-43 antigen rial ES-31 serine protease. These inhibitors also showed was primarily recognized by serum antibodies in relapse decreased bacterial growth in axenic culture and inhibition cases. Further it has been shown that ES-6 and ES-20 have is further confirmed by decreased amount of ES-31 serine serodiagnostic potential in latent infection and lymphnode protease in culture filtrate. In human macrophage culture, tuberculosis respectively. The later is observed at elevated highly inhibitory pefabloc, 1,10 phenanthroline and isoniazid level in weak immune patients. Immunomonitoring for inhibited infectivity of virulent as well as avirulent M. presence of antigen in TB patients under ATT, showed tuberculosis bacilli to macrophages. The enhancement of that ES-31 antigen assay was useful in determining the mycobacterial cell growth in the presence of mycobacterial effectiveness of therapy and patient’s compliance. Further ES-31 serine protease while 90% inhibition (as observed by a cocktail of ES antigens SEVA TB ES-31, ES-43 and EST-6 decreased CFU count) in the presence of anti ES-31 serine (containing 38 kDa and 41 kDa protein antigens) and their protease antibody was observed showing the importance affinity purified antibodies were explored and found useful of enzyme for entry and multiplication of bacilli and thus in diagnosis of pulmonary and extra pulmonary tubercu-

123 S24 Ind J Clin Biochem

may be used as drug target. Orlistat may have therapeutic M.tb in cell cultures and to differentiate from non tuber- use in isoniazid resistant cases and in suspected cases of culous mycobacteriae (NTM). Further our findings support tuberculosis. Immunofluorescence study using FITC labelled the concept that SEVA TB ES-31 as extracellular protease of anti ES-31 antibody conjugate showed the presence of ES- M. tuberculosis which is readily accessible target of these 31 serine protease on mycobacterial cell surface and thus impermeable organisms may be useful in rapid screening found to be a sensitive and specific biomarker to confirm of potential antitubercular drugs.

INNOVATIVE TECHNOLOGY TO QUANTITATE SERUM PROTEINS COMPONENTS

Dr. D.M. Vasudevan Principal (Retd), Amrita Institute of Medical Sciences, Kochi

In blood, some of the proteins are present in much lower technology will have the advantages of both turbidimetry concentrations, while others are seen in much higher and nephelometry. Such an instrument should have the concentrations. Designing a single instrument for mea- following capacities: 1) Automatic selection of nephelom- suring all the serum proteins of diagnostic significance is etry and turbidimetry, depending on the concentration of a big challenge to the in vitro diagnostic (IVD) industry. protein present in the solution. 2) Eliminate time consum- The turbidimetry and nephelometry had been developed ing calibration process. 3) Should have a semi-automatic a few decades ago. Nephelometry was not very popular or automatic platform. 4) Should avoid orminimize errors. because of the cost of the instrument. Turbidimetry was 5) Should be able to work with minimum quantity of not very popular due to lack of sensitivity, although having reagents and serum samples. 6) Should detect prozone very good specificity. It is well established that turbidim- phenomenon automatically and should warn the opera- etry offers high linearity and nephelometry offers greater tor. 7) Should have the QC function. 8) Should be able sensitivity. For the same reason, turbidimetry is used to to quantitate all serum proteins. 9) Should have good determine proteins present in high concentrations, while sensitivity and specificity. 10) Instrument should be simple nephelometry is used to determine proteins present in to operate. 11) Cost should be affordable to ordinary labo- lower concentrations. Therefore, it will be ideal if we could ratories. Recently such an instrument hasbeen launched in combine turbidimetry and nephelometry. This innovative the Indian market.

GLUTATHIONE PEROXIDASE 1 POLYMORPHISM IN ANEMIC CHILDREN

Dr. Neelima Singh Department of Biochemistry, G.R. Medical College, Gwalior

Anemia is associated with low antioxidant enzyme de- C allele and 57.21 % for A allele, as compared to 43.48 ficiency especially Glutathione Peroxidase. GPX exist in % for C allele and 23.48 % for A allele of age matched 6 isomeric forms out of which GPX1 Single Nucleotide control group. The genotyping heterozygous frequency Polymorphism is found to be associated with many types of G/C & G/A genotype in GPx 1 polymorphic marker in of anemia. In our study, the determination of the allelic anemic children were (33.83 % & 54.72 % respectively). frequency and phenotype of a common polymorphism in The statistical analysis of data showed that the difference Se-dependent glutathione peroxidase1 (GPX1) was observed in the genotypic frequency between the two groups is in Anemic populations. The study has been carried out in significant for G/C genotype (P<0.05) and for G/A geno- 40 anemic cases and 40 control subjects. The genotypes type (P< 0.001). No significant association between allele encoding G/C and G/A are distributed according to the frequency and risk to get fatal was evident. Erythrocyte Hardy–Weinberg relationship. The allelic frequency of GPx GPX activity was determined and significant differences 1 polymorphic marker in anemic group was 40.54 % for were obtained between the genotypes when compared

123 Ind J Clin Biochem S25

with control group (P<0.001) It can be concluded that Keywords: GPX 1, Anemia, SNP. G/C & G/A genetic variation appear to compromise the defense against oxidative stress in red blood cells along with other antioxidants in blood.

ECONOMICS AND MANAGEMENT OF QUALITY ENHANCEMENT IN A CLINICAL LABORATORY … PERSONAL EXPERIENCE AS A CONSULTANT

Dr. Uday Donde Consultant for Accreditation, Director. UXL Advisor. Mumbai

The Director of every Clinical Laboratory is supposed to pation in the external quality assurance programs (EQAP), be judicious in using the available resources and quality are the economic constraints of this process. conscious in reporting on clinical material. An enthusiastic Validation of analytical procedures and calibration of the Laboratory Director may be keen on getting his laboratory major equipments used in the analysis are a primary re- accredited from a regulatory authority; but so may not be quirement for laboratory work. Although the cost involved the case with several others. Clinical Laboratory Accredita- in this is marginal, it increases precision and avoids repeat tion is defined as a process of evaluation of the labora- experimentation. Once this is established, quality enhance- tory by a qualified and competent third party to ensure ment does not become a big issue. In-house training to conformity with pre-defined criteria. The EQAS (external the staff is imperative, and is difficult to compute in terms quality assurance scheme) service provider’s grade the of its economics. performance of a laboratory based on criteria applied at a national / international level. Accreditation is optional in As a Consultant to diagnostic laboratory services’ groups some countries, mandatory in others. in India, I have given consultations to large level and mid level laboratories on these issues for a decade. Advice on Quality enhancement begins with self appraisal of all the laboratory reorganization, minimum / maximum capital activities carried out in a laboratory covering all the special- investment needed for quality enhancement; handling izations. These activities may range from giving instructions the component of in-house training, incentives to staff to the patient, error-free specimen collection and transport etc. have been the crucial issues. Document preparation to the analytical station, reliable and speedy analysis and and update; record keeping / archival is cumbersome but eventually giving the right report to the right person for necessary. A systematic approach to laboratory transforma- appropriate action. This unbroken chain demands constant tion and the economics involved in quality enhancement monitoring and use of standard operating procedures. will be discussed during the presentation. The possibility Designing these procedures is a matter of logistics and of converting this approach to “webinars” will also be implementing them is a skill. Adopting internal quality discussed. control (IQC) measures at an affordable cost, and partici-

123 S26 Ind J Clin Biochem

SYMPOSIA V : DIABETES, ENDOCRINE & HEMATOLOGICAL DISORDERS

ROLE OF VITAMIN D SUPPLEMENT THERAPY ON GLYCEMIC CONTROL IN VITAMIN D DEFICIENT TYPE 2 DIABETIC MELLITUS PATIENTS

Charu Agarwal, Sadhana Marwah, Bindu Kulshreshtha, Anubhuti Dr Ram Manohar Lohia Hospital, PGIMER, New Delhi

Aims and objectives: Vitamin D is a secosteroid that is patients were followed up serially and grouped into cases either synthesized in the skin by the action of sunlight or and controls. Thirty patients were given cholecalciferol ingested in the diet. It plays a significant role in immu- (cases) and other 30 served as control. All patients were nity and insulin secretion. Vitamin D is thought to affect followed up. At the end of three months, the values of pancreatic β cell function (insulin synthesis and secretion) fasting and post prandial sugar, HbA1c and Vitamin D were and immune response. The mechanism by which vitamin D compared with those measured at baseline month. might act on insulin secretion is suggested by significant Results: The group of diabetic patients supplemented with rise in cytoplasmic calcium levels observed for vitamin D vitamin D showed a significant improvement in post pran- stimulated secretion of insulin by islet cells. Therefore, dial glucose levels (p= 0.016). Fasting glucose and HbA1c this study is planned to look for the changes in glycemic values also showed a decline but it was not statistically levels in diabetic patients after supplementing Vitamin significant. D in South Asian population since South Asians are the population with a high prevalence of Vitamin D deficiency Conclusion: Supplementation of vitamin D for three months and Type 2 Diabetes mellitus. improved levels of postprandial sugar levels in vitamin D deficient T2DM in the present study. However a larger Materials and methods: A total of 60 patients of T2DM number of study population with longer duration of study with HbA1c ranging between 7% - 8.5% and vitamin D levels would be helpful to determine the role of vitamin D in less than 75nmol/l were taken from the diabetic clinic. On glycemic control in T2DM patients. the basis of computer generated random number table,

EFFECT OF HORMONE REPLACEMENT THERAPY ON LIPID PROFILE

Indu Verma*, Promila Jindal, Rama Sofat, Vandana Chopra Department of Biochemistry* and Gyneacology, Dayanand Medical College & Hospital, Ludhiana

Background: Women with premature menopause are at for three months, while group II B patients received me- higher risk of vascular complications due to thrombogen- droxy-progesterone acetate daily for last ten days of each esis and atherogenesis. Hyperlipidemia is one of the major month. Lipid profile was done before operation, after one risk factor of atherosclerosis so the effect of hormone and fourth month postoperatively. Results were analysed replacement therapy (HRT) was studied on lipid profile by t test & p value in women after surgical menopause. Results & Discussion: The mean levels of total lipids, cho- Materials and Methods: 100 women after total abdominal lesterol, triglycerides, low density lipoproteins (LDL), very hysterectomy (TAH) with bilateral salpingo-oopherectomy low density lipoproteins (VLDL) levels increased after one (BSO) were divided into two groups. Group I (control) month of hysterectomy in all the three groups. The increase comprised of 50 women who received no HRT. Group in levels of total lipids, cholesterol, LDL, VLDL continued II (study) subdivided into II A and II B with 25 patients in control group till the end of the study while in study each. After one month of surgery all the women of group group this trend was reversed at the end of the study. II received 0.625 mg of conjugated equine-estrogen daily Hormone replacement therapy decreased total lipids, cho-

123 Ind J Clin Biochem S27

lesterol, triglycerides, LDL, VLDL by 12.2% and 9.6%, 8.97% is after surgical menopause where changes are abrupt. and 1.31%, 17.7% and 15.4%, 8.24% and 7.59%, 15.8% and 20.0% Correlation between cardio vascular disease & post meno- in group II A and group II B respectively. On comparing pausal HRT clearly indicate that addition of progestin does the results in control and study group, the differences are not appear to attenuate the cardio protective effects of statistically significant although no significant difference post menopausal estrogen therapy. was observed between subgroup II A and II B. Keywords: HRT, surgical menopause, lipid profile. Conclusion: HRT plays an important role in combating complications of natural menopause but more important

TO STUDY THE GLUCOSE6PHOSPHATE DEHYDROGENASE DEFICIENCY STATUS

Narendra Kumar Gupta1, Matendra Kumar2, Department of Pathology1, ESIC Model Hospital Cum Occupational Disease Centre, Indore, Department of Pathology2, ESIC Hospital, Adityapur, Jamshedpur

Objective & Background: Glucose-6-Phosphate Dehydroge- substrates Glucose-6-Phosphate & NADP to give NADPH, nase (G6PD) defiency is one of the most common inherited this NADPH decolorizes the blue colored indophenol dye hemolytic disorders reported & studied. It affects 400 (DCPIP) in presence of PMS. It leaves behind color which million people world wide. In India about 4 lacs children is due to hemolysate. The rate of reaction is proportional are born annually with this disease. G6PD difiency is sex to enzyme activity of G6PD present in erythrocytes. The (X) linked trait affecting males, while females are carriers time required for decolorization is inversely proportional & asymptomatic. Individuals develop hemolytic episodes to enzyme activity in the hemolysate. on exposure to oxidant stress such as viral & bacterial Result: The results were analyzed & data compiled. Our infections, certain drugs (antimalarials, sulfonamides, ni- study involves 500 persons (300 males & 200 females). trofurantoin, aspirin, vitamin K), metabolic acidosis & on G6PD deficiency was detected in 45(15%) males & 10(5%) ingestion of fava beans (favism). females with total of 55(11%). Methods: Test was done on the patient who was referred Conclusion: Prevalence of G6PD deficiency in India is var- to laboratory for detection of G6PD status. The study was ies from 0-28%. Our study follows the Indian trend. But done for 6 years from 1st July 2006 to 30th June 2012. significant number of females (5%) is also G6PD deficient Measurement of G6PD was done by qualitative enzyme in our study. assay. In this test G6PD present in hemolysate acts on

TREND OF LIVER ENZYME RECOVERY POST LIVE DONAR LIVER TRANSPLANT

Dr Neelima Verma1, Dr Vivek Vij2, Dr Vishal Kumar Chourasia3, Dr Rajiv Tangri4, Dr Neena verma5, Dr Sonia Vij6 1Deptt of Clinical Biochemistry, Fortis Hospital, Noida. 2Liver Transplant & GI surgery, Fortis Hospital, Noida. 3Liver transplant & GI surgery, Fortis Hospital, Noida. 4Histopathologist & Lab Head, Fortis Hospital, Noida. 5Sr Pathologist, Fortis Hospital, Noida. 6Consultant Pathologist, Fortis Hospital, Noida.

Objective: Serial monitoring of liver enzymes is very crucial aberration from the pattern can be better & timely may for monitoring recovery of patients after living donor liver be dealt with to avoid post transplant complications. transplant because in these cases partial liver is utilized Material & Methods: All the end stage liver disease patients which shows a characteristic enzyme pattern during re- undergoing live donor liver transplant (n=22) admitted generation process. Therefore it is important to have in the year 2012 at Fortis Hospital, Noida were included understanding of normal liver enzyme recovery pattern in the study. Serial monitoring of levels of SGOT, SGPT, with respect to liver regeneration so that any pathological

123 S28 Ind J Clin Biochem

SAP & SGGT was done for up to three weeks & trend Conclusion: Monitoring of liver enzymes and knowledge of was observed. normal patterns of recovery is crucial in patient manage- ment following live donor liver transplant. Results: Out of 22 patients of liver transplant 16 patients were male & 6 patients were female. Age varied from Keywords: Liver transplant, Serum glutamic oxaloacetic 33-62 yrs for male patients & 46 - 58 yrs for female transaminase, Serum glutamic pyruvic transaminase, Se- patients. There was only one child (9yrs old male) who rum gamma glutamyl transferase, Serum alkaline phos- underwent liver transplant. In the patients who had nor- phatase. mal recovery SGOT & SGPT tend to peak after 48-72 hrs Abbreviations: LT – Liver Transplant, SGOT – Serum glu- of transplant then tend to decline. SAP & SGGTP tend to tamic oxaloacetic transaminase, SGPT – Serum glutamic rise by the end of first week & peak at around 8-10 days pyruvic transaminase, SGGT – Serum gamma glutamyl then remain elevated for another 3-4 days before com- transferase, SAP – Serum alkaline phosphatase. ing to normal after 14-18 days. Any deviation from this pattern marked presence either of biliary complications, rejection or sepsis.

UNMASKING OF ALPHABETA DOUBLE GENE DELETION THALASSAEMIA TRAIT BY HBA1C ANALYSIS

Rinchu Loomba, R. Chawlaand Vikas Loomba* Department of Biochemistry, *Department of Medicine, CMC & Hospital, Ludhiana

Thalassemia Minor (Trait) is usually a benign condition that We did Hb electrophoresis using cellulose acetate strips produces only mild anemia and usually goes undiagnosed. and got a value of 6.2% for HbA2, which was confirmed Here we present an unusually silent case of Thalassemia by affinity column chromatography. However, the RBC trait which was unraveled by incidental HbA1c analysis indices in CBC did not conform withThalassemia trait, and bylow pressure cation exchange chromatography on a Bio were suggestive of only of a mild iron deficiency. These Rad Dia STAT analyser. anomalous observations prompted us to go for DNA analy- sis. The ARMS-PCR and southern blot analysis reported The haemolysate of this 27 years old married medical co-inheritance of bothα and β gene deletion leading to student was run as a control to establish the laboratory the RBC indices which were not as reduced as expected reference ranges of HbA1c by dia STAT analyser. She was in classical β thalassemia trait. non-diabetic with no family history of Diabetes Mellitus. The HbA1c value was alarmingly high at 22.0% and 23.2% in The family screening later confirmed the younger sibling two consecutive runs. HbA1c values of all other controls to be carrying similar thalassemia trait and helped in his run in parallel were well within the expected range. This selection of a non-carrier marital partner. led us to think of a possibility of presence of some other Keywords: HbA1c, Hb, Hemolysate, Thalassemia trait. Hb variant in the hemolysate.

COMPARATIVE STUDY OF HAEMATOLOGICAL PARAMETERS AND CELL INDICES WITH LEVEL OF G6PD IN PATIENTS OF AVBRH

Dr Ruchir Khare, Dr A W Meshram Department of Biochemistry, J N Medical College, Sawangi (M), Wardha

Object of study: Most studies of G6PD have focused on G6PD in nucleated cells have been largely overlooked. To the molecular characterization of different G6PD variants, the best of our knowledge no study has been taken to the pathophysiology of G6PD-deficient erythrocytes, and study the hematological indicators with normal or near the hemolytic aspect of G6PD deficiency. The roles of normal level of G6PD and its relation with morphological

123 Ind J Clin Biochem S29

effects on red blood cells. Results and observations: Various biochemical parameters such as G6PD and hematological parameters such as MCV, Aim and objectives: Study of hematological parameters MCHC, MCH, hematocrit, RBC& WBC count and hemo- and their correlation with G6PD concentration in patients globin were recorded and will be discussed with suitable with morphological changes of RBCs with various level statistical methods. of G6PD. Conclusions: G6PD greatly affect the red cell morphology Material and Methods: 30 patients suffering from vari- in deficient condition. Changes in other cell number and ous skin disorders & 30 healthy control were taken from structure is also studied and their relationships with other AVBRH and 2ml blood is collected after informed consent. hematological parameters were analyzed. Along with G6PD Biochemical investigations are done. G6PD, hematological some hematological parameters and cell structure can give parameters have been evaluated using suitable biochemi- better assessment of future events like hemolysis. cal procedures. Various cell morphology studied with the help of peripheral smear.

HYPOMAGNESEMIA, LIPID PROFILE AND GLYCOSYLATED HAEMOGLOBIN IN TYPE 2 DIABETES MELLITUS PATIENTS

Dr. Supriya, Dr. Shrabani Mohanty, Dr. Roopa Murgod, Dr. D.S. Raghavendra Department of Biochemistry, VIMS&RC, Bangalore

Objective: Hypomagnesemia is a common feature in haemoglobin were significantly higher in cases as com- patients with type 2 diabetes mellitus. It is known to pared to controls. cause vasoconstriction & endothelial injury, making it an A significant negative correlation between serum mag- atherogenic risk factor. Our objective of this study was nesium and glycosylated haemoglobin was observed. to investigate the effect of magnesium on lipid profile & Also, negative correlation was observed between serum also on glycosylated haemoglobin. magnesium and lipids such as total cholesterol, triglycer- Material & Methods: This study was conducted in 100 ides, VLDL, LDL, whereas a positive correlation was seen subjects, out of which 50 were type 2 diabetes mellitus between magnesium & HDL. patients (cases) and 50 were non diabetic healthy subjects Conclusion: From our study we were able to conclude (controls). serum Magnesium, Glycosylated haemoglobin that magnesium has a significant role to play in serum and serum lipid profile were measured by using the auto lipid profile. Hypomagnesemia results in elevation of total analyzer Beckman coulter DXC600. cholesterol, TG, LDL, VLDL, whereas it lowers HDL levels. Results: Our study revealed that, serum magnesium and Hypomagnesemia also results in elevation of glycosylated HDL cholesterol were significantly lower, whereas other haemoglobin levels. lipids like total cholesterol, TG, LDL, VLDL & Glycosylated

URIC ACID AS A BIOMARKER IN THE DIAGNOSIS OF SYNDROME OF INAPPROPRIATE ANTIDIURETIC HORMONE SECRETION SIADH

Dr Sutirtha Chakraborty Consultant & Chief, Clinical Biochemistry, Peerless Hospital & BK Roy Research Centre, Kolkata

Syndrome of inappropriate antidiuretic hormone secretion results from inappropriate, continued secretion and/or (SIADH) is a common cause of hyponatremia which itself action of antidiuretic hormone (ADH) despite normal or is the commonest electrolyte abnormality. The syndrome increased plasma volume, which results in impaired wa- is defined by the hyponatremia and hypo-osmolality that ter excretion. Till date SIADH does not have consensous

123 S30 Ind J Clin Biochem

diagnostic criteria and remains more of a diagnosis of control population (n=45) we included those hospitalized exclusion. But diagnosis is of great importance because the patients who had depletional hyponatremia as a result of management is more of fluid restriction rather than normal dehydration, diuretics etc. saline therapy used in cases of depletional hyponatremia Result analysis showed that the SIADH group of patients like dehydration. Interestingly it has been suggested that had a significantly lower serum UA as compared to the serum and urinary Uric Acid (UA) levels can be useful other control group. All patients with SIADH had serum UA biomarkers in the diagnosis of SIADH. levels of less than 3 mg/dl. FE-UA was significantly higher Considering this background, a study was done to assess in SIADH patients (> 12%) than the control group. the importance of serum uric acid and renal handling of Our study shows that serum UA and FE UA are simple, urate through Fractional Excretion of Uric Acid (FE-UA). cheap and readily available investigations that can help The cases (n=45) included all clinically suspected SIADH in the diagnostic approach of hyponatremia leading to patients admitted to our hospital with hyponatremia, identification of SIADH. who met the inclusion and exclusion criteria. Serum and urinary UA was assayed on an Olympus 480 platform. As

123 Ind J Clin Biochem S31

SYMPOSIA VI : INSTRUMENTATION IN LAB PRACTICES & ANIMAL EXPERIMENTS

IMMUNOFIXATION ELECTROPHORESIS AN IMPORTANT AID TO THE DIAGNOSIS, MANAGEMENT AND PROGNOSIS IN MULTIPLE MYELOMA PATIENTS THE CMC LUDHIANA EXPERIENCE

R. Chawla, M. Joseph John, Rinchu Loomba, Maria Thomas Christian Medical College & Hospital, Ludhiana (Pb)

Immunotyping in multiple myeloma (MM) patients is of only in two of our cases. One case of biclonal prolifera- utmost importance considering the wide variation in the tion of immunoglobulins (IgG and IgA) was also reported. response to therapy and prognosis in these patients. The Kappa light chains were more common with IgG type of data generated from 44 multiple myeloma cases using immunoglobulins, whereas IgA immunoglobulins associated immunofixationelectrophoresis (IFE), estimation of im- more with the lambda light chains (55.6%). munoglobulins, free light chains, β2-microglobulins, serum Position of the M-band on SPE was of little significance protein electrophoresis (SPE) and Bence Jones Proteins either for the type of immunoglobulin or occurrence of (BJP) was analyzed with the following results. the free light chain disease (FLC), although M-band near IFE showed that IgG (62.1%) is the most common type of β-globulins was, more often than not, found to be IgA immunoglobulin in MM patients, followed by IgA (31.0%). or FLC. IgM type of multiple myeloma was rather rare, occurring FLC disease was diagnosed in 34.1% of the MM patients and was associated with significantly increased risk for Total Total% kappa Lambda k% L% nephropathy and poorer prognosis. Free lambda light IgG 18 40.9 14 4 77.8 32.2 chains were more common (60%) compared to free kappa IgA 9 20.5 4 5 44.4 55.6 light chains and were associated with significantly higher IgM 2 4.5 2 0 100.0 0 mortality rates. β2-microglobulins also predicted a poorer FLC 15 34.1 6 9 40.0 60 outcome. BJP showed a moderate sensitivity (68%) but 44 very high positive predictive value (95%). IFE is an important tool for immunotyping of multiple Kappa 26 59.1 myeloma for management and prediction of outcome in Lambda 18 40.9 these patients.

CURRENT CLINICAL CHALLENGES & SOLUTIONS IN AUTOMATED BIOCHEMISTRY ANALYSIS IN INDIA

Atanu Kumar Dutta Demonstrator (Lecturer), Department of Biochemistry, IPGMER, Kolkata

Clinical Biochemistry or Laboratory medicine has a great term total laboratory automation was coined by Sasaki in influence on clinical decisions and 60-70% of the decisions 1967 which was realized in 1990s with the advancement in on admission, discharge and medications are based on labo- robotics and development of “belt” system for delivering ratory results. The birth of laboratory automation should specimens to analytical systems. Other types of automa- be dated in the 1950s when the first continuous flow tion technologies include modular laboratory automation autoanalyzer (Technicon, “Autoanalyzer”) was developed and workcell/workstation automation. These two types of to measure Creatinine and uric acid. This was followed by automations are being used in most of the Indian labora- use of flame photometry and Coulter counter (1957). The tories which couple preanalytical and postanalytical work

123 S32 Ind J Clin Biochem

done by manual methods to automated analysis of clinical chemistry analysis from ingested drugs and other factors specimens. For example in IPGMER, Kolkata Clinical Bio- which contributes significantly to Clinico-Biochemical dys- chemistry Laboratory most of the pre and post analytical correlation. Postanalytical automation also include sample work is done manually whereas routine tests are performed storage stations and autovalidation softwares which may in two random access autoanalyzers and emergency and help to perform additional tests requested by the clinician special tests are performed in four semiautoanalyzers. As in a later period and maintain uniform standards in patient preanalytical phase constitutes about 60% of the time in reporting throughout the day. Use of analytical algorithms a diagnostic process and can account for 46-68% of the for reflex testing, for example assay of fT4 when TSH total errors according to various studies, the absence of is 4-10 mU/L or <0.02 mU/L can dramatically reduce the preanalytical automation which includes an aliquotter and sample load and prevent unnecessary strain over limited automated centrifuge in most of the Indian laboratories resources. These types of algorithms may also help in the imposes major problems in minimizing preanalytical errors. practice of evidence based medicine in the resource poor Postanalytical phase accounts for 19-47% of total errors settings in India and better utilize the laboratory resources which comes from improper handling of laboratory data for diagnosis, cure and prognosis. Another major drawback and also contributes to prolonged turnaround time. Al- of the Indian scenario is that point of care testing is only though laboratory softwares are usually used in most of sparingly used, though automated POCT instruments have the large Indian laboratories to prepare and store patient been validated to be highly helpful in timely clinical deci- reports, the practice of using laboratory information sys- sion making. There is also a huge gap in the research and tem and hospital information system is not very common education in the field of Laboratory Medicine in India. in India at least in the Government hospitals. Hospital Even postgraduate courses in Biochemistry do not provide information system also provide laboratory professionals ample scope for training in state of the art automation with the access to electronic patient records which may system and data handling. Collaboration with industry may help them to find out possible interference in clinical offer an olive branch.

PROTECTIVE EFFECT OF DOCOSAHEXAENOIC ACID ON ALUMINUMMEDIATED METABONOMIC CHANGES IN RAT URINE; A PROTON NUCLEAR MAGNETIC RESONANCE STUDY

Devesh Kumar Joshi1, Manisha Choudhary1, Sudhanshu Mishra1, Sandeep Tripathi1, Abbas Ali Mahdi2, Raja Roy2 1Department of Biotechnology, Nims Institute Engineering & Technology, NIMS University, Jaipur 2Department of Biochemistry, K.G. Medical University, Lucknow 3Centre of Biomedical Magnetic Resonance, SGPGIMS Campus, Lucknow

Background: Aluminum (Al) is responsible for dialysis Results: Metabolic profile of Al treated rat urine showed dementia and Alzheimer’s disease. Patients with chronic significantly decreased levels of citrate, creatinine, allantoin, kidney disease on regular haemodialysis treatment are trans-aconitate, succinate and increased acetate concentra- exposed to Al. Docosahexaenoic acid (DHA) has been tion. While DHA treated rats exhibited significantly reversed used extensively as a modulator of brain, kidney and liver. changes near to control. These results demonstrated Aim of the present study was to investigate therapeutic the impairment urea cycle in kidney, which was further potential of DHA on nephrotoxicity particular. reinstated by clinical chemistry and histopathological ob- servations. Methodology: The toxic effects of aluminium (Al) have been studied in 90-days AlCl3 orally treated male albino rats Conclusion: On the basis of results it may conclude that (n = 7) and simultaneously, protective efficacy of 100mg DHA ameliorates Al induced nephrotoxicity and cellular / kg b.w. DHA was tested using 1H NMR spectroscopy- changes in rats. Furthermore, DHA can be suggested for based metabonomic profile of rat urine and histopathol- the intervention to kidney failure patients. ogy of kidney.

123 Ind J Clin Biochem S33

ACBI/CMC EQAS QUO VADIS ?

F.S. Geethanjali, R. Selvakumar Department of Clinical Biochemistry, Christian Medical College, Vellore

Objective: The objective of this retrospective study is to Results: The awareness of Quality assurance in India has evaluate the usefulness of the ACBI/CMC EQAS program in tremendously increased and is reflected by a 100% increase the overall performance of Indian Laboratories as well as in the number of participants over the past ten years. The individual labs in different categories over the years. It is randomly selected labsshowed decreased OMVIS which is the journey of our National EQAS over the past decade. indirectly proportional to their performance.

Method: The awareness ofthe Indian labs towards qual- Conclusion: It was during this past 10 years, lot of changes ity assurance is evaluated based on the number of took place in the ACBI/CMC EQAS, i.e., change over form participants, the response in submitting results, annual bovine sera to human sera, liquid samples to lyophilised evaluation and the trend in methods selection. Similarly samples, introducing immunoassay program etc. This study few labs form each category such as lab size (number of shows how the Indian laboratories have improved their parameters done), type (public/private) and few labs at performance as well as their awareness towards the QA- random were selected and their performance is evaluated concept. It is very encouraging to note that the concept over the past 10 years. of EQAS is well accepted and appreciated by majority of laboratories and thus paving the way towards international standards.

NEEDS AND CHALLENGES OF TRANSFORMATION OF LABBASED ASSAYS INTO DIAGNOSTIC KITS: AN EXPERIENCE WITH DIAGNOSTICS KITS FOR FERTILITY ASSESSMENT

M. Ikram Khatkhatay National Institute for Research in Reproductive Health (ICMR), J.M. Street, Parel, Mumbai

Hormones, being key bio-regulatory molecules, their mea- validated and then transformed into kits by providing in- surements have assumed a position of pivotal importance dividual components of the assays in ready-to-use format. in diagnosis of endocrine disorders, which are prevalent in The kits contained antibody coated plates in dry-format India. Development of inexpensive home-grown diagnostics besides lyophilized standards, concentrated enzyme labels kits and their availability to masses is therefore needed. and substrate-chromogen which are required to be recon- Besides hormones, indigenous assays for other biomarkers stituted or diluted appropriately before use. The shelf-life developed at laboratory level generally remain at prototype of kits was determined by accelerated stability studies. level only. We share herewith the challenges and modalities Subsequently, second party validation was performed by of transforming indigenously developed ELISA for estrone analyzing coded samples received from endocrinologist and glucuronide (E1G), pregnanediol glucuronide (PdG), the comparing results with clinical parameters. For third party principal metabolites of estradiol and progesterone in validation, an external agency provided the kits to clinicians urine into commercial mode. Antisera raised against E1G who independently analyzed samples and compared with and PdG were checked for its specificity by checking its standard parameters. We conclude that data on stability crossreactivity with compounds present in urine. Enzyme studies along with second and third party validations of labels were prepared by conjugating E1G and PdG to developed kits are mandatory steps; and facilitate their peroxidase. ELISAs developed using these reagents, were commercialization.

123 S34 Ind J Clin Biochem

TOURNIQUET APPLICATION TIME DURING PHLEBOTOMY AFFECTS BIOCHEMICAL TESTING

Monika Gupta1, Dharamveer Yadav2, Sandhya Mishra2 and Praveen Sharma3 1Department of Biochemistry, Mahatma Gandhi Medical College & Hospital; 2SMS Medical College, Jaipur; 3All India Institute of Medical Sciences, Jodhpur

Introduction: Venous blood sampling is usually performed protocol of commercially available kits. using a tourniquet to help locate and define peripheral Results: ALT concentration was significantly high in samples veins to achieve successful and safe venepuncture. In de- collected after application of external pressure for 3min veloping countries like India, there is lack of understanding (P<0.05) and 6min (P<0.01); Alkaline phosphatase, LDH, about good laboratory practices and inadequate training Creatine kinase, Total Cholesterol, Triglyceride, Total Protein, to phlebotomists, compelling them to make errors during Albumin and Calcium concentrations were significantly high phlebotomy. The improper venous accesses or prolonged in samples collected after application of external pressure venous stasis created by tourniquet application will result for 6min (P<0.05) as compared to samples collected with- in collection of unsuitable blood sample. out venous stasis. There were no significant alterations in Methods: The present study was conducted at one of rest biochemical analytes concentrations (Glucose, Urea, the tertiary care hospital at State Capital to find out the Creatinine, AST, Uric Acid, Phosphorus, Sodium, and Potas- effect of tourniquet application time on 17 common bio- sium) even after 6 min venous stasis. chemical analytes in 20 fasting healthy adult volunteers. Conclusion: Venous stasis from tourniquet placement dur- Sequential venepunctures were performed by a single ing venepuncture should be minimized, as it accounts for expert phlebotomist either without venous stasis or fol- spurious and significant variations for several biochemical lowing the application of standardized external pressure analytes. of 60 mm Hg using a sphygmomanometer for 1, 3 and 6 minutes. Biochemical parameters were analyzed on Fully Keywords: Tourniquet, Venous Stasis, Biochemical Analytes, Automated Analyzer Olympus AU 400 by using standard Phlebotomy

ANTIHYPERLIPIDIMIC AND ANTIOXIDANT ACTIVITY OF STEVIA REBAUDIANA BERTONI IN TRITON INDUCED HYPERLIPIDEMIC RATS

Poonam Yadav, Madhvi Bhardwaj, Purnima Kumari, Sandeep Tripathi, Sandeep Kumar Plant Tissue Engineering & Metabolomics Research Lab, Department of Biotechnology, NIET, NIMS University, Jaipur

Background: Leaves of Stevia rebaudiana Bertoni have Results: Oral administration of the S. rebaudiana (200 mg/ been extensively used as a sweetener in foods and bever- kg body wt.) exhibited a significant reduction (P<0.01) in ages for diabetics and obese people due to their potent serum lipid parameters like total cholesterol, triglycerides, sweetener stevioside. It regulates blood sugar and prevents low density lipoprotein (LDL), very low density lipopreotein hypertension. Multiple complications are reported to be (VLDL) and increase in high density lipoprotein (HDL) in common in those parts of the world where dietary fat hyperlipidemic rats. Moreover, lipid peroxide levels and intakes are high. antioxidant status (SOD, CAT, GPx and GSH) were found to be reversed in treated with extract. Methodology: In present study; we evaluated the hyper- lipidemic effects of leaves extract of S. rebaudiana (100, Conclusion: Our results demonstrated that (200 mg/ kg 200 and 400 mg/ kg for 30 days) in titron induced hy- body wt) of S. rebaudiana possessed significant antihy- percholesteromic rats. Moreover a comparison was made perlipidemic activity and hence it could be a potential between the action of S. rebaudiana leaves extract and herbal medicine as adjuvant with existing therapy for the a known antihyperlipidemic drug simvastatin (10 mg/kg treatment of hyperlipidemia along with antihyperglycemic body wt.). & antihypertensive activity. Moreover it also ameliorates increased rate of lipid peroxidation.

123 Ind J Clin Biochem S35

NEPHROPROTECTIVE EFFECT OF KARISALAI KARPAM POLYHEDRAL FORMULATION IN CARBON TETRACHLORIDE CCL4 INDUCED NEPHROTOXICITY IN RATS

R. Ganesan*, B. Ramyaa**, G. Pramod Reddy*, P. Sathyarajeshwaran* and S. JagaJothi Pandian* *Siddha Central Research Institute (SCRI), Arumbakkam, Chennai, **SASTRA, University, Thanjavur

Alcoholic extract of aerial parts of Karisalai Karpam in the cellular alterations were scored as Nil, +, ++ and +++ Tween 80 was evaluated for nephroprotective activity for No, Mild, Moderate and Severe damage, respectively. It in Wistar albino rats. All the animals were weighed and was observed that in the Group II (P<0.05) and Group III randomly divided into four groups comprising of six rats (P<0.01) there was significant reduction in BUN, Creatinine in each group. The kidney damage was induced by i/m as compared to that of Group IV. In the sections of kidney injection of Carbon tetrachloride (ccl4) @ 1.5 ml/kg on 6th obtained, there was mild degree of damage in group III day onwards in group II, III and IV. Group I and IV were (+) followed by Group II (+) as compared to damage in kept as normal (Saline) and nephrotoxic control group, Group IV (+++). There were no changes in the sections respectively. Whereas, Group II and III were treated with obtained from Group I, which was normal control. From Karisalai Karpam @ 45 mg/kg and 90mg/kg orally respec- the results obtained it can be concluded that Karisalai tively, for 10 days. The blood samples were estimated Karpam may be effective as nephroprotector. for biochemical parameters. The kidney were collected, Keywords: Karisalai Karpam, Nephroprotective effect, Car- weighed and processed for histopathology. The kidney bon tetrachloride. sections were examined for histopathological changes and

123 S36 Ind J Clin Biochem

SYMPOSIA VII : INVITED LECTURES

ANTIOXIDANT FOOD AND DRINKS FOR YOUR HEALTH AND PREVENTING DISEASES

Amitava Dasgupta Professor of Pathology and Laboratory Medicine University of Texas Health Sciences Center at Houston

Free radicals such as superoxide anion, peroxide radical, important role in aging process. Many fruits and vegetables hydrogen peroxide, hydroxyl radical as well as singlet are rich in natural antioxidant and best defense against oxygen are produced during normal cellular metabolism oxidative stress is to eat healthy meals including generous especially mitochondrial respiration which is essential for servings of fruits (at least 80 gm a day) and vegetables burning glucose as a fuel. However, body has can neutral- (80 gm or more a day) each day. Tea and coffee are ize these highly reactive free radicals through vitamins also good source of antioxidant while coffee is richer in such as vitamin C, vitamin E, beta-carotenoids as well as antioxidant than tae. Chocolate is also rich in antioxidant antioxidant enzyme such as superoxide dismutase, cata- but chocolate is full of calories. Red wine is also a rich lase and peroxidase. Oxidative stress is generated when source of antioxidant because flavonoids present in grape body’s antioxidant defense to detoxify free radicals is skin especially resveratrol is present in higher quantities inadequate. If not neutralize free radicals can damage vari- in red wine than white wine. Resveratrol can provide ous organic molecules essential for life including proteins, defense again cardiovascular diseases and can reduce lipids, carbohydrates and DNA. In humans oxidative stress risk of developing Alzheimer’s disease. Although certain is related to diseases including cancer, cardiovascular dis- vitamins are antioxidants, current research indicates that eases especially arthrosclerosis, diabetes, psychiatric illness taking such fat soluble vitamins as supplements can cause and even neurodegenerative diseases such as Parkinson’s more harm than good. disease and Alzheimer’s disease. Oxidative stress plays an

GENOMICS OF HUMAN HEART FAILURE: PUTATIVE THERAPEUTIC TARGETS

Hari S. Sharma Institute for Cardiovascular Research, Department of Pathology, VUmc University Medical Center, Amsterdam, The Netherlands

Tetralogy of Fallot (TF) is a common form of cyanotic with RVB of age matched controls. Total RNA was isolated, congenital heart disease (CHD) where primary correction at subjected to cDNA synthesis and eventually processed for a young age is the treatment of choice. Main features in the preparation of labelled cRNA probes which were used tetralogy of Fallot (TF) include right ventricular hypertrophy to hybridise Affymetrix U95A expression chips containing (RVH) and failure of which underlying molecular mecha- ~12,600 genes in separate experiments. After scanning nisms are not yet understood. We hypothesise that various and computer analysis of the expression data in RVB of forms and stages of CHD in children are associated with ToF patients was compared to the baseline expression of a number of characteristic novel genes (e.g., cell surface respective genes in control. Data were considered signifi- receptors, signalling molecules and transcription factors cant when the mean expression changed >2 fold. About etc) and a subset of these genes would predict the etiol- 200 genes showed up and down regulation pattern by ogy and molecular phenotype of cardiovascular defect. We setting a threshold of 2 and above. Data analysis revealed performed microarray analysis on RVB from 4 patients who that certain group of Genes involved in cell proliferation underwent primary correction for the ToF (mean age 0.8 and growth were down regulated where as extra cellular yr) and compared the expression profile of various genes matrix genes were up-regulated. Myocardial expression

123 Ind J Clin Biochem S37

of fibrosis markers assessed by video-image analysis and in TF-1 patients. Furthermore, molecular pathway analysis corrected for hypertrophy showed significantly increased revealed complex gene interaction in both angiogenesis interstitial fibronectin (p<0.01), total collagen (p<0.01) and and fibrosis pathways. Quantitative RT-PCR analysis of collagen subtype I (p<0.01) and III (p<0.01) in both TF1 and mRNAs revealed enhanced expression pattern in TF-2 as TF2 as well as peri-vascular fibronectin (TF1; p<0.01, TF2; compared to C2. We conclude that RVH is associated with p<0.05) and total collagen (TF1;p<0.01) as compared to enhanced myocardial fibrosis and apoptosis and thereby respective controls. TIMPS showed down regulated pattern. diminishing cardiac function in patients with TF. Further- Immunostaining of VEGF in cardiomyocytes was increased more, up-regulation of genes encoding VEGF seems key (p<0.01) in patients with TF. Morphometric analysis revealed event contributing to right ventricular hypertrophy and enhanced vascular density (p<0.05) with unchanged wall failure associated with stunted angiogenesis and targeting thickness and enlarged myocyte cross sectional areas these genes and their products may serve as potential (p<0.01) with linear correlation (r=0.92, p<0.01) with the age therapy for patients with heart failure.

123 S38 Ind J Clin Biochem

SYMPOSIA VIII : MALADIES OF REPRODUCTIVE SYSTEM & TOPICS ASSOCIATED WITH PREGNANCY

SIGNIFICANCE OF SERUM URIC ACID LEVEL AND ITS RELATION TO OUTCOME IN HYPERTENSIVE DISORDER OF PREGNANCY

Hariom Sharma*, Nikunj Modi*, Amit Maheshwari* *Department of Biochemistry, Govt. Medical College, Bhavnagar

Abstract: Pregnancy induced hypertension is a multisys- group, with a p value of 0.003. Proteinuria is also pres- temic syndrome and a leading cause of maternal and fetal ent in the case group in the patients of preeclampsia and morbidity and mortality. This study comprised 125 cases eclampsia but it is not present in the normal healthy preg- of pregnancy induced hypertension (PIH) and 125 healthy nant woman. But Blood urea and Serum Creatinine levels pregnant women with normal blood pressure as a control in both cases and controls is within normal limits. This group. They were primarily diagnosed for PIH by clinical finding rules out any renal disorder. In the present study examination followed by Biochemical investigations and we found that Patient with high serum uric acid levels are their fetal outcomes were observed. Mean Uric acid level at risk of worst fetal outcome as compare to those who was 3.8 ± 1.0 mg/dl in the normotensive group and 7.4 ± have normal serum uric acid levels. Thus high serum uric 2.2 mg/dl in (PIH) case group, with a p value of < 0.001. acid level in patients of pregnancy induced hypertension Serum urea was 26.5 ± 9.1 mg/dL in the normotensive can be used as marker for fetal outcome. group and 23.4 ± 6.6 mg/dl in PIH case group, with a p Keywords: Fetal Outcome, pregnancy, Pregnancy induced value of 0.002. Serum creatinine was 0.7 ± 0.4 mg/dL in hypertension (PIH), Uric acid. the normotensive group and 0.6 ± 0.2 mg/dl in PIH case

STUDY OF IRON PARAMETERS IN PREGNANCY INDUCED HYPERTENSION

Dharmveer Yadav1, Jairam Yadav2, Monika Gupta3, Sandhya Mishra1 and Praveen Sharma4 1SMS Medical College, Jaipur. 2Ashvini Naval Hospital, Kolaba, Mumbai. 3Mahatma Gandhi Medical College & Hospital, Jaipur. 4All India Institute of Medical Sciences, Jodhpur.

Background: Pregnancy induced hypertension (PIH) is a as admitted in wards of Obstetrics and Gynaecology in disorder of unknown etiology unique to pregnant woman. Mahila Chikitsalaya, Ajmer were included in the study. Incidence of PIH in India is about 7-10%. It is estimated that Fasting blood samples were collected from anticubital vein upto 30% of perinatal deaths are related to hypertensive using asceptic technique were analyzed for estimation of disorders of pregnancy. Since the condition is preventable, serum Iron, UIBC, TIBC, Ferritin, Total Bilirubin and blood it is essential to diagnose the disease at early stage so Hemoglobin by standard methods. that proper medical care can be instituted in time. Results :- Mean values of serum Iron, Ferritin, TIBC, UIBC, Objective: In the present study an effort has been made Total Bilirubin and blood Hemoglobin in study and control to define more precisely the levels of various Iron me- groups were 141.89±45.58 μg/dl v/s 92.08±19.60 μg/dl; tabolism related biochemical parameters i.e. serum Iron, 146.01±64.29 ng/dl v/s 52.02 ± 22.29 ng/dl; 309.64±46.12 μg/ Ferritin, TIBC, UIBC, Total Bilirubin and blood Hemoglobin dl v/s 255.38±32.92 μg/dl; 250.71±43.67 μg/dl v/s 221.57±36.91 in PIH patients and to establish their significance in early μg/dl; 0.95±0.13 mg/dl v/s 0.91±0.10 mg/dl respectively. diagnosis, prognosis and management of the disease. The mean serum Iron and Ferritin levels were significantly higher (p<0.001) and mean TIBC and UIBC were signifi- Material and Methods: 75 primigravida women ranging cantly decreased (p<0.001) in PIH subjects as compared in age from 20-35 years of which 50 with PIH and 25 to healthy controls. with normal pregnancy attending Antinatal clinic as well

123 Ind J Clin Biochem S39

Conclusion: Increased levels of serum Iron and ferritin diagnosis will help in better prognosis and management and decreased levels of serum TIBC, UIBC, Bilirubin and of the disease. blood hemoglobin may help in early stage diagnosis of Keywords: Pregnancy induced hypertension Iron, UIBC, PIH. Since PIH is a preventable condition, its early stage TIBC, Ferritin, Bilirubin, Hemoglobin.

THE INFLUENCE OF GENERAL MATERNAL WEIGHT ON SERUM ALPHAFETOPROTEIN AND FREE BETAHUMAN CHORIONIC GONADOTROPIN LEVELS DURING THE SECOND TRIMESTER OF PREGNANCY

Diganta Dey, Abhijit Banerjee, Parbati Banerjee, Arnab Paul and Ratnamala Ray Dept. of Biochemistry, Ashok Laboratory Clinical Testing Centre Pvt. Ltd., Kolkata

Objective: Maternal serum alpha-fetoprotein (AFP) and There was a markedly inverse relationship between mater- free beta-human chorionic gonadotropin (beta-hCG) levels nal weight and serum AFP and free beta-hCG levels. The are useful biomarkers for antenatal screening of Down median AFP and free beta-hCG level for women weighing syndrome, Trisomy 18 and open neural tube defects. The ≤ 50 kg were respectively 90.4% and 57.4% higher than objective of this study was to investigate the relationship the median level for women weighing 70 kg or more. The between maternal weight and serum AFP and free beta- overall variation in AFP (p = 0.17) and free beta-hCG (p hCG concentrations. = 0.69) levels were significant between these two weight groups. The reference interval (2.5 to 97.5 percentile) for Methodology: The study included 70 normal singleton AFP and free beta-hCG were obtained 5.56 – 136.37 ng/ pregnancies (mean age: 31.45 years) without any congenital mL and 4.41 – 283.55 IU/mL respectively. anomalies, were screened using AFP, free beta-hCG and unconjugated estriol between 16 and 20 weeks of gesta- Conclusion: Maternal weight may affect the AFP and free tion, during January 2010 to July 2012, using enzyme linked beta-hCG concentrations during the second trimester. Heavi- immunosorbent assay (ELISA; AFP-hCG-uE3 AccuBind VAST er pregnant women have lower median values, perhaps ELISA Kit, USA), followed by the PregSafe prenatal risk as a result of a diluting effect of larger blood volume. A assessment software. revision of the reference interval for AFP and free beta-hCG considering the body weight might improve in assessing Results: The mean maternal weight was 59.75 ± 18.02 kg risk of poor outcome, especially in heavier women. in studied pregnant women during the second trimester.

EXCESSIVE FLUORIDE IN DRINKING WATER REDUCES THE QUALITY OF SEMEN

Dushyant Singh Chauhan1, Vivek Pratap Singh1, Sudhanshu Mishra1, Sandeep Tripathi1, Sandeep Kumar1, Surabhi Tomar3, Anurag Tomar2 National Referral Centre for Fluoride Poisoning in India, 1Department of Biotechnology, Nims Institute of Engineering & Technology; 2Department of Paediatrics, 3Department of Gynaecology, Nims Medical College, NIMS University, Jaipur

Background: Humans are exposed environmentally to fluo- healthy age matched controls were selected from the non ride. Fluoride accumulates in the tissues, bones and body fluoride region (<1.5ppm) of the Rajasthan. Semen and fluids. It has been reported that fluoride accelerates free blood samples were collected for assessment of semen radical production and increased lipid peroxidation and quality (motility, count, viability) and testicular hormonal further it may lead to reproductive toxicity. In this study profiles (Follicle stimulating hormone, lutinizing hormone, we attempt to evaluate serum fluoride levels and semen inhibin-B and prolactin). Moreover, lipid peroxide and an- quality in fluoride exposed population. tioxidant status was also evaluated.

Method: We recruited 189 individuals (30 to 50 Years), Results: Analysis of semen samples revealed that sperm who were residing in high fluoride region (>2.5 ppm) and count was significantly (p>0.01) decreased while the pro-

123 S40 Ind J Clin Biochem

lactin level was increased markedly (p>0.05) in subjects. Conclusion: On the basis of results it may safely conclude On the other hand, LH and FSH levels were found to be that, fluoride increases prolactin which may alters the reduced in subjects as compared with the healthy control. secretion of LH and FSH. Perturbed LPO and antioxidant Moreover, insignificant changes observed in the concentra- is the marker of oxidative burden. Moreover, it may be tion of inhibin. Increased LPO and decreased antioxidants due to the alteration in pathophysiology of hypothalamic (Superoxide dismutase, catalase, glutathione peroxidase axis. Further study is needed for explanation of the above and Glutathione) were observed in cases. problem.

BIOCHEMICAL ASPECTS OF CREATININE CLEARANCE AS A MARKER FOR DIAGNOSING AND ASSESSING THE SEVERITY OF PREECLEMPSIA

Dr. Satya Prakash*, Ms. Neha Sharma** Dr. (Mrs.) Puja Kumari*** Dr. Ajit Kumar* *Department of Biochemistry, Rajendra Institute of Medical Sciences (RIMS), Ranchi, Jharkhand **Department of Biochemistry, Shri Ram Murti Smarak Institute of Medical Sciences (SRMSIMS), Bareilly ***Department of Obstetrics and Gynecology, Rajendra Institute of Medical Sciences, Ranchi, Jharkhand

Background: Creatinine is the metabolic waste product Results: The present study was undertaken to analyse the resulting from the breakdown of creatine (which remains relationship between creatinine clearance and pregnant as creatine phosphate in muscle), and is usually produced women. The results were: at a fairly constant rate by the body (depending on muscle Creatinine clearance mass). Serum creatinine though is a simple test and is the a. In normal non-pregnant cases, the mean was 102.63 most commonly used indicator of renal function a rise ml/min. in blood creatinine levels is observed only with marked b. In normal 1st trimester pregnant cases, the mean was damage to functioning nephrons. Therefore this test is not 127.3 ml/min. suitable for detecting early stage kidney malfunctioning. c. In normal 2nd trimester cases, the mean was 142.1 ml/ A better estimation of kidney function is given by the min. creatinine clearance test. In view of the aforementioned d. In normal 3rd trimester cases the mean clearance was controversial literature, it was decided to evaluate the 128.7 ml/min. relation-ship between the renal changes (creatinine clear- e. In mild Preeclamptic cases, the mean was 127.81 ml/ ance) in pre-eclamptic, eclamptic, preeclamptic toxeaemic, min. non pregnant and normal pregnant women. f. In moderate Preeclamptic cases, the mean was 103.81 Methods: The present investigations were undertaken to ml/min. study creatinine clearance in normal non-pregnant women, g. In severe Preeclamptic cases, the mean fall down to Pregnant women in different trimesters, Preeclamptic 77.68 ml/min. toxaemic patients and eclamptic patients admitted in h. In eclamptic cases, the mean found was around 71.98 Rajendra Institute of Medical Sciences, Ranchi during the ml/min. period from June 2008 to September 2009. Control group The difference between each group is highly significant consisted of 15 normal non-pregnant women, 10 cases except when control and mild cases were compared in first trimester, 30 cases in 2nd trimester, 20 cases in (p>0.005). 3rd trimester, 60 cases of Preeclamptic toxaemia and 20 cases of eclampsia. Creatinine clearance measurements Conclusions: Clearance fall gradually in cases from mild to were done by Enzymatic method and modified Jaffe’s severe Preeclamptic cases. But the difference in fall of the kinetic method. clearance values were not highly significant in mild and moderate cases when compared to normal 3rd and 2nd trimester values. However, it is a good index to gauge the renal status in these patients.

123 Ind J Clin Biochem S41

MATERNAL NUTRITION AND OFFSPRING OBESITY/ADIPOSITY: INDIAN SCENARIO

Dr. Sadanand S. Naik, Dr. C.S. Yajnik K.E.M. Hospital Research Centre, Pune, India The global epidemic of childhood obesity is a major public vegetables, fruit and milk was a strong determinant of health issue. The factors acting during early life have long fetal growth. “Early life nutritional exposures (intrauter- term effects on health and the process is programmed ine and neonatal) play a prominent role in programming in-utero. Fetal life is critical window for programming the susceptibility in adult life to chronic diseases, such as a later body fatness is termed as ‘fetal programming’. obesity, CVD, DM, osteoporosis. Asian Indians have a unique phenotype characterized by We along with International collaborators have created increased abdominal obesity and visceral fat despite low conceptual bridge between some of the most important BMI. The study done in K.E.M. Hospitral Research Centre, global public health challenges of our time: obesity/adipos- Pune, supports the idea that the risk of excess energy ity, fetal/early life origins of adult disease, and prevalent intake is accentuated in those who are born small, i.e. and persistent micronutrient malnutrition. 2/3rd of Indian catch up growth is influencing cardio vascular risk. mothers are vitamin B12 deficient and their off-springs are The study carried out at 6 villages near Pune, has shown short and thin, but have more body fat. This is associated that the mothers size, food intake, physical activity and with high prevalence of Insulin Resistance, presumably circulating concentrations of nutrients and metabolites dur- leading to epidemic type-2 DM among adults in India. ing pregnancy are important determinants of baby’s size Therefore, the prenatal environment which programs sub- and body composition at birth. The rural Indian newborn sequent obesity/adiposity provides a potential therapeutic babies who are 800 gms lighter than the White Caucasian target. The study in this field to readily translate into babies have almost similar sub-cutaneous-skin fold-thickness improved interventional strategies is under progress. which suggests that the ‘Thin-fat-phenotype’ of Indians originates in-utero. The frequency of intake of green leafy

MATERNAL VITAMIN D LEVELS AN ALARM FOR PREECLAMPSIA

Victoria Job and Reshma Rajhu Department of Clinical Biochemistry and Community Medicine, Christian Medical College, Vellore Introduction: Preeclampsia is one of the major causes for Methods: A nested case-control design. Cases and controls maternal and fetal mortality and morbidity. In developing were selected from patients registered in the clinic. Serum countries, the incidence of preeclampsia ranges from 1.8- sample is stored at – 20ºc for all pregnant women. Cases 16.7% and in India 2-8%. Some potential risk factors are ma- - 45 Pregnant women who were diagnosed to have preec- ternal age, race, exposure to sun. 1,25 dihydroxy VitaminD lampsia. Controls - 90 i.e. two subsequent pregnancies with has immunoregulatory functions and is a potent inhibitor similar gestational age and parity. VitaminD was analysed of Th1 cell proliferation, which is correlated with spontane- using an Electrochemiluminescentimmunoassay. ous abortions. Normal pregnancy is a Th2 cell dominant Results: The VitaminD levels were equally distributed process. Recent studies have shown controversial findings among the cases and controls. The mean levels among on the association between VitaminD and preeclampsia. the cases was 21.47 and controls 23.57 ng/ml. There is Baker reported a 4 fold risk of severe preeclampsia with no significant difference between the groups (p value is < 20 ng/ml level of VitaminD in midgestation. Powe found 0.29). The matched pair analysis revealed no association no association between first trimester VitaminD levels and between VitaminD deficiency and preeclampsia. subsequent preeclampsia. Conclusion: Low socioeconomic strata had 61% less chance Aim: To determine the association between maternal of developing preeclampsia and muslim women had a serum VitaminD levels in early pregnancy and the risk of 8.1% higher risk. preeclampsia in a community health hospital.

123 S42 Ind J Clin Biochem

SYMPOSIA IX : FUNCTIONAL DISORDER

TO CORRELATE THE NUTRIENT INTAKE WITH SOCIOECONOMIC CHARACTERISTICS AMONG THE MALE AND FEMALE CARDIAC PATIENTS

Archana Singh1, Meenakshi Verma Department of Biochemistry, Institute of Home Science, Dr. B.R. Ambedkar University, Khandari, Agra

Abstract: Cardiovascular disease today is one of the most tor contributing to high prevalence. Multistage stratified leading cause of mortality in both developed and develop- systematic sampling techniques were used for selecting ing countries. The incidence of hypertension, heart attack 100 samples and an interview schedule was evolved to and atherosclerosis has increased multi-fold. Today, life is collect the information regarding socioeconomic profile, a race in which each person tries to compete with other dietary pattern and specific information regarding this in terms of wealth, status, success, reputation and several disease. In this study we found the consumption of high such ideals through fair and unfair means. This accompa- fat diet was revealed as the major contribution by cause nying tension and worries, the craving for the cigarette, of CHD. The mean BMI significant and positively correlated this compelling peg, the leisurely or hectic made of travel with Vitamin C among the male CHD patients whereas in and overweight, all drag the person towards a disease. significant and positively correlated with Vitamin C among The present study was conducted to know the prevalence female CHD patients. The dietary intake between male and of CVD between male and females and major risk fac- female were highly significant.

ROLE OF HORMONAL THERAPY FOR REDUCING PROSTATE SPECIFIC ANTIGEN LEVEL, IN THE MANAGEMENT OF CARCINOMA PROSTATE

Dr. Ashika. Mudilikulam Sukumaran, Dr. Jagathlal Palliparambil Chandrasekharan, Dr. Rajeev Kavalakara Raghavan

Background: Prostate cancer is one of the most common 2012, falling in the age group of 40 to 90 years. The total cancers in men, predominantly a disease of the older age number of samples analyzed were 400 from 200 patients. group. Even though the exact etiology of the disease is The PSA values of these patients before and after treat- unknown, it takes many years for the cancer cells to grow. ment were studied. The patients were grouped into two Over the past 10-15years, there have been a number of categories, one who received hormonal therapy and the significant advances in the management of prostate car- other who had undergone treatment modalities, other than cinoma. Hormonal Therapy is found to be having a major hormonal therapy. The first group enlisted 133 patients and role by reducing serum prostate specific antigen level. second group enlisted 67 patients. All the samples were LHRH analogues, Anti androgens and Diethyl Stilbestrol analyzed by Chemiluminescence method. are effectively used as hormones for this current purpose. Results: PSA values were assessed in both the groups, It involves manipulation of the endocrine system, through before and after receiving the treatment. The reduction exogenous administration of these hormones to control in PSA in the first group was 82.7% whereas it was only the progression of the disease process. 68.7% for patients belonging to the second group. The Aim of the Study: To highlight the effects of hormonal results analysis showed remarkable reduction in PSA titer therapy in reducing PSA level, so as to make the treat- by the influence of hormones. The values were found to ment of prostatic cancer more effective. be statistically significant (p value<0.05).

Materials and Methods: This is a retrospective analysis of Conclusion: From this study, we found that, hormonal serum PSA levels and its influence on hormonal therapy, therapy, in the management of carcinoma prostate, is managed in the Department of Oncology, Amrita Institute effective in reducing the serum PSA levels so as to pro- of Medical sciences and Research Centre. The period of vide a control over the pathogenesis of the disease and study of patients ranges from January 2000 to January a better prognosis in long run.

123 Ind J Clin Biochem S43

EFFECT OF SEASONAL CHANGES ON THYROID PROFILE OF REPRODUCTIVE AGEGROUP FEMALES IN NORTH INDIA

Basu A, Seth S, Agarwal H K, Bhadra J, Kulshrestha M R, Kumari A

Objective: To study the contrast between thyroid profiles significantly higher than that (11.20, sd = 26.48) in winter during summer and winter in reproductive age-group (p=0.0004). Serum T3 also demonstrated significant con- females. trast (p<<0.0001) between summer (mean = 147.89, sd = 62.25) and winter (mean = 130.36, sd = 84.38). In the Methods: Analysis of retrospective data of female patients year-wise analysis, significantly higher mean TSH in summer (20 – 40 years) undergoing thyroid profile test in the De- was found during 2009 (p<0.0001) and 2011 (p<0.0001); partment of Biochemistry, PGIMS Rohtak was performed. significantly higher mean T3 in summer was found during Serum TSH was measured with IMRA (immunoradiometric 2010 (p=0.0006) and 2011 (p=0.005); while 2009 winter assay; normal = 0.3 - 5.0 microIU/ml). Serum T3 (normal (mean) T4 level was significantly higher than in summer = 70 - 200 nanogram/dl) and T4 (normal = 5.5 -13.5 mi- (p=0.0068). crogram/dl) was measured with RIA (radioimmuno assay). Pooled data for 3 consecutive summers (April, May, June Conclusion: The overall results clearly demonstrate interest- of 2009, 2010, 2011) and 3 consecutive winters (November, ing seasonal variations in thyroid profiles of reproductive December, January of these 3 years) were compared. age-group women of Northern India. Further prospective study of thyroid profiles for hyper, hypo and euthyroid Results: Thyroid profile data of (n=1261) patients were groups, taking into account the pathological basis is rec- studied, including (n=632) in summer and (n=629) in ommended. winter. Overall analysis for 3 consecutive years showed that mean TSH level (16.78, sd = 29.04) in summer was

CORELATION OF GLEASON’S SCORE, PROSTATE SPECIFIC ANTIGEN PSA & PROSTATIC ACID PHOSPHATSE PAP LEVELS IN ADENOCARCINOMA OF PROSTATE

Gaurav Karve*, A.K. Tandon** *P.G. Student, **Professor, Department of Biochemistry, BVDUMC, Pune

Prostate cancer is the 2nd most common cause of cancer immunoassay (CLIA) & chemiluminescent microparticle im- deaths among males. 90% cases are adenocarcinomas of munoassay (CMIA). prostate. It is slow growing & largely non-symptomatic dis- Out of 423 subjects screened, 35 (8.27%) were diagnosed ease. Study was conducted to identify co-relation between as prostate adenocarcinoma. They were divided into 3 the Gleason scores and concurrent serum prostate-specific groups depending on their Gleason score. The mean PSA antigen (PSA) and prostate acid phosphatase (PAP) values. & PAP values in Low grade cancer group (n=12) were 37 The Gleason’s scoring system is a histopathological grading ± 8.15 & 3.0 ± 0.39 respectively; in Intermediate grade scheme for prostatic adenocarcinoma. PSA and PAP are cancer group (n=13) they were, 123 ± 62.8 & 4.83 ± 1.26 widely accepted serum markers for prostatic diseases. respectively, and in high grade cancer group (n = 10) 435 The study was conducted on 420 elderly males above 40 ± 150 & 8.79 ± 2.98 respectively. years of age, freshly diagnosed with prostatic carcinoma This indicates a clear positive correlation between the in Bharati Hospital Research Centre (BHRC). They were three parameters. The PSA & PAP levels are elevated in subjected to clinical examination, Digital Rectal Examina- prostatic adenocarcinoma with PSA showing significant tion (DRE), Sonography, serum PSA & PAP estimation & increase and better correlation with Gleason score as Biopsy. PSA & PAP were estimated using chemiluminiscence compared to the PAP levels.

123 S44 Ind J Clin Biochem

ROLE OF ABCB6 IN MITOCHONDRIAL BIOGENESIS IN CANCER CELL LINES

Pamela Christudoss Dept. of Clinical Biochemistry, CMC, Vellore

Objective: ABCB6 is a mitochondrial transporter that determined by real time quantitative reverse transcription regulates porphyrin biosynthesis. ABCB6 expression is polymerase chain reaction and quantitative polymerase up regulated in hepatocellular cancer cell lines, but the chain reaction. significance of this up regulation to cancer is not known. Results: ABCB6 expression was up regulated in all the 3 In the present study, we investigated 1) ABCB6 expres- cell lines. NRF-2and T-fam expressions were up regulated in sion in 3 human cancer cell lines- Hep G2 (liver), Hek 293 both Hep-G2 and K-562 over expressing cell lines as com- (kidney) and K562 (erythroblastoma). 2) Expression of the pared with vector cell lines. A significant up regulation of key mitochondrial transcription factors NRF-2 (nuclear re- the expression of some of the downstream mitochondrial spiratory factor) and T-fam (mitochondrial transcriptional genes was observed in the HepG2 cell lines only. factor) in these cell lines. 3) Expression of the downstream mitochondrial genes COX-1 and 2, ND-1, 5, 6 and ATPase Conclusion: The study has shown a correlation between 6 which are regulated by NRF-2, and T-fam in the HepG2 T-fam and NRF-2 activity and the expression of some and K562 cell lines. proteins encoded by mitochondrial DNA in the Hepatoma cell lines. These data suggest the transcriptional control Method: ABCB6 over expressing cells were cultured in of the biogenesis marker in mitochondrial biogenesis in Dulbeccos modified Eagle’s medium (DME medium) sup- the hepatocellular carcinoma cell line. plemented with glutamine, and penicillin. ABCB6, NRF-2, T-fam and the downstream mitochondrial genes were

STUDY OF SERUM CREATINE KINASE AND LACTATE DEHYDROGENASE IN HYPOTHYROIDISM

Dr. Reena R.1, Dr. H.V. Shetty2 1Post-Graduate Student, 2Professor and Head of the Department, Department of Biochemistry, Rajarajeshwari Medical College, Bangalore.

Background: Hypothyroidism is one of the most com- were taken as controls. Biochemical estimations of T3, T4, monly occurring thyroid disorders, worldwide. Deficiency and TSH by ultra sensitive chemiluminescence, serum CK of thyroid hormones slows down the metabolic function and serum LDH by enzyme kinetic method in Transasia and in muscles this results in pain on exertion and delayed EM 360 autoanalyzer were carried out. reflexes. 30-80% of hypothyroid patients presents with Results: 99% of the study subjects were females in the muscular symptoms varying from myalgia to true myopa- age group of 30-35 years with mild muscular symptoms. thy. To confirm the involvement of muscles, biochemical Serum CK (25-192 U/L) and serum LDH (80-227U/L) were tests such as estimation of serum Creatine Kinase (CK) well within normal reference range in both cases and a sensitive marker of muscle damage and serum Lactate controls. Dehydrogenase (LDH) a general marker of tissue damage are useful. The aim of this study was to assess the involve- Conclusion: In hypothyroidism the serum CK or LDH levels ment of muscles in hypothyroid patients by estimating may not be elevated when muscular symptoms are mild; serum CK and LDH levels. early diagnosis and timely intervention may also be one the reasons for the normal levels of serum CK and LDH Materials and Methods: 50 hypothyroid patients (irrespec- in these patients. tive of the duration of the disease) with history of muscle symptoms were taken as cases and 50 healthy adults with Keywords: Thyroid disorders, Creatine Kinase (CK), Lactate normal TFT attending medical OPD of RRMCH, Bangalore, Dehydrogenase (LDH).

123 Ind J Clin Biochem S45

ASSOCIATION OF INSULIN RESISTANCE WITH ENDOTHELIAL DYSFUNCTION IN METABOLIC SYNDROME

Sandhya Lal1, Vandana Saini1, Jayashree Bhattacharjee1, MK Bhatnagar2 Department of Biochemistry1 & Department of Medicine2, Lady Hardinge Medical College and Shrimati Sucheta Kriplani Hospital, Delhi

Background: Insulin resistance is strongly associated with Results: Statistical analysis was done using SPSS version different components of metabolic syndrome (MetS) includ- 19. The levels of ET-1, a potent vasoconstrictor and NO a ing obesity. The role of insulin resistance in the determina- potent vasodilator were found to be significantly higher tion of endothelial dysfunction is not known. (p<0.05) in cases as compared to controls. Insulin resis- tance was found to be significantly higher in cases as Objective: To study the effect of insulin resistance on compared to controls (p<0.05). There was no significant endothelial dysfunction markers endothelin-1 (ET-1) and association between insulin resistance and the endothelial nitric oxide (NO) in MetS in New Delhi. dysfunction (p>0.05). Methodology: We conducted a hospital based case-control Conclusion: Association between insulin resistance and study. It included 46 MetS cases (IDF 2006) attending medi- endothelial dysfunction is not statistically significant. Both cine OPD of LHMC and SSKH, New Delhi and 47 healthy insulin resistance and endothelial dysfunction are associated volunteers. Fasting plasma Glucose level were estimated independently with Metabolic Syndrome. on clinical chemistry auto analyzer & Plasma insulin levels were estimated by chemiluminiscent based immunoassay Keywords: Metabolic Syndrome, Insulin Resistance, En- (CLIA). HOMA-IR was calculated for insulin resistance. dothelial Dysfunction. Plasma ET-1 levels were estimated by ELISA. Plasma NO levels were measured by modified Griess reaction.

SERUM ADENOSINE DEAMINASE AND ITS ISOENZYME ACTIVITY IN ASTHMA

Sharma J, *Menon BK, **#Vijayan VK, Bansal SK Departments of Biochemistry, *Respiratory allergy and applied immunology, and **Pulmonary medicine, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi. #Bhopal Memorial Hospital and Research Centre, Bhopal, Madhya Pradesh Asthma is a chronic inflammatory disorder of airways and its isoenzymes (ADA1, ADA2) in serum. The data was causing variable airflow obstruction. In asthma, adenosine analyzed statistically and a p < 0.05 was considered as levels increase; this may then interact with mast cells, significant. lymphocytes and monocytes-macrophages via its receptors The results show that in asthma the total ADA activity to cause inflammation. Adenosine is converted to inosine decreased significantly in group II and III patients in com- by Adenosine deaminase (ADA, E.C.3.5.4.4). The increase parison to healthy controls (p< 0.0001). The difference was in adenosine may be due to decrease in ADA activity. also significant when group I was compared with group II There are two isoenzymes of ADA in serum; ADA1 and and III (p<0.05). There was a significant difference in ADA1 ADA2. The objective of this study was to examine the and ADA2 activities between groups II, III in comparison changes in activity of ADA and its isoenzyms in serum of with control (p< 0.0001). ADA activity in group I did not asthma patients and correlate them with the severity of differ from control significantly. The FEV1% has a positive airflow obstruction. correlation with the serum ADA activity (r =0.6127). The study included 45 patients of bronchial asthma classi- We conclude that ADA and its isoenzymes activity decrease fied into three groups viz. mild persistent (group I), moder- with increase in severity of airway obstruction or vice-versa, ate persistent (group II) and severe persistent (group III) which may play an important role in airway inflammation and 15 healthy controls. Blood was collected and serum and possibly in pulmonary injury in asthma. separated, followed by determination of activities of ADA

123 S46 Ind J Clin Biochem

SYMPOSIA X : INVITED LECTURES

ROLE OF C677T POLYMORPHISM IN METHYLENETETRAHYDROFOLATE REDUCTASE MTHFR GENE IN NORTH INDIAN INFERTILE MEN

Abbas Ali Mahdi*, M.K. Ahmad*, S. R. Hussain*, H. Naqvi*, $S.P. Jaiswar, S.N. Shankhwar# *Molecular Cell Biology Lab., Department of Biochemistry & Departments of $Obstetrics & Gynaecology and #Urology, King George’s Medical University, Lucknow

Background: Failure or difficulty in conceiving a child is spermia, astheno-zoospermia, astheno-terato-zoospermia, surprisingly becoming a common worldwide problem. Half oligo zoospermia, oligo-astheno-teratto zoospermia (OAT), of these cases are due to male factors with defect in sperm oligo-astheno zoospermia, oligo-terato zoospermia, normo (1 in 15 men) being the single most common cause. Also zoospermia, necro zoospermia and 364 fertile north Indian about 60–75% of male infertility cases are idiopathic, since men as healthy controls by using PCR-RFLP technique and the molecular mechanisms underlying the defects remain Chi-Square test for statistical analysis. unknown. DNA methylation is crucial for spermatogenesis Results: The average MTHFR 677CC, 677CT, 677TT geno- and high methylenetetrahydrofolate reductase (MTHFR) type frequencies of total infertile men were 70.17%, 24.17%, activity in adult testis than other organs in mouse, signifies 5.65% in cases and 75.27%, 21.7%, 2.74% in healthy controls, its critical role in spermatogenesis. According to recent respectively. The average frequency of the MTHFR 677T findings there is a correlation of epigenetic regulation of allele was 23.39% in the infertile men as compared to 16.34% several imprinted genes with disturbed spermatogenesis of healthy controls and the difference was established and fertility. Consequently any change in the MTHFR statistically significant. gene sequence can modify the spermatogenesis including transmission of infertility to the carriers. Conclusions: Our findings indicate that there is an asso- ciation of C677T in the MTHFR gene polymorphism with Materials and Methods: To analyze the distribution of the male infertility, indicative of this polymorphism might be single nucleotide polymorphism (SNP) C677T in the MTHFR a genetic risk factor for male infertility in north Indian gene in 637 north Indian infertile patients with azoo- population.

ADDRESSING LAB CHALLENGES THROUGH AUTOMATION & PROCESS EXCELLENCE

Arun Raizada, Aditi Gupta, Kamini Vinayak, Vijay Kher, Naresh Trehan Medanta-The Medicity, Gurgaon

The Laboratory Medicine department supports various clini- patients and doctors. Most laboratories are trying to cope cal departments by providing valuable information about up with these changing demands through automation, the health of a patient as pertaining to the diagnosis, consolidation, increasing use of Information Technology treatment, and prevention of disease. Clinical laboratory and improved processes. Automation solutions are now testing thus plays a crucial role in patient care and manage- available at almost all phases of laboratory testing namely ment. The Indian healthcare scenario is undergoing major – pre-analytical, analytical and post analytical. Over the changes with reference to clinical care, consumer/ patient years the analytical platforms have evolved from simple behavior, service standards etc. The clinical laboratory is colorimeters to fully automated integrated analyzers. facing numerous challenges today is terms of pressure Integration (combined chemistry and immunoassay ana- to improve quality, reduce medical errors, improve TAT, lyzer) have helped improve lab productivity and operator handle increasing test volumes and complexities with efficiency by simplifying and consolidating user interface, space, budget and staff shortage. Laboratories across the maintenance schedules, inventory management, quality world are trying to find ways and means to meet these assurance and management, consumable usage etc. the operational challenges and deliver quality services to the biggest advantage has been less dependence on skilled

123 Ind J Clin Biochem S47

manpower and reduced stress levels on existing staff. This of the above mentioned advantages would fail. Automa- has led to reduced TAT including add on tests and lesser tion well suited to your lab needs when coupled with sample volume requirements resulting in improved patient process solutions like Lean can bring about path breaking management by physicians. Improved lab productivity is improvements in the overall operational efficiency of your helping improve the brand value of the hospital, increase lab. These Lean solutions can provide immense benefits revenue and savings by reducing average length of stay to patients, physicians, employees as well as the hospital for patients and improving patient care standards. How- management by improving workflow, creating standard ever the best of automation alone is not enough. If the work systems, introducing error proofing systems and processes around automation are not streamlined most introducing performance metrics.

ACCUMULATION OF LEAD IN VEGETABLE CROPS ALONG MAJOR HIGHWAY IN KOLKATA, INDIA

K. Goswami, Adelene Basu Department of Pathology, ESIC Hospital & ODC (EZ), DH Road, Joka, Kolkata

Introduction: Exposure to lead (Pb) through food, water, Result: The experiment was set up in blocks at random or contaminated air has adverse health impacts that are with four replicates. The results showed that carrot and particularly severe in children. The problem associated beet were the most Pb-tolerant species, while the others with the disposal of sewage water and municipal waste were very Pb- sensitive. The decreasing order for Pb ac- is important aspects of water pollution which should be cumulation in the vegetables crops was: carrot > spinach considered in the context of wider environmental problem. > tomato > cabbage > cauliflower > bringle > ladies finger Use utilization of waste water for irrigation purposes has > chilly > beet. Taking into account the Pb allocation in increased over the past years, as this waste contain high plants, in order was root > stems > leaves > edible parts. amount of heavy metals, like Pb. Although carrot translocated the lowest Pb amount in the edible part, such level exceeded the legally acceptable or Objective: To determine lead levels in the environment permitted limit. and foods around Kolkata City in India and assess with reference implications human health hazards. Conclusion: These results indicate that there is considerable risk of lead poisoning from eating some foods from these Methodology: The amount of Pd in selected samples of sites. Also, the concentration of Pb in all the vegetables vegetables obtained directly from Warmian-Masurian pro- tested even after washing exceeded the permissible limit cedures. Concentration of analyzed compounds after dry recommended by WHO / FAO. Also, recommendation for mineralization of biological material and organic phase waste water utilization for irrigation if the content of toxic extraction (APDC / MIBK) has been determined using elements is higher than the permissible limits is reduced atomic absorption spectrophotometry (AAS) in a Perkin- considerably. Elmer, AAS 2380.

VERIFICATION PROTOCOL IN THE LABORATORY: FIRST STEP OF STANDARDIZATION AND HARMONIZATION OF THYROID FUNCTION TEST

Barnali Das Consultant, Biochemistry & Immunology, Kokilaben Dhirubhai Ambani Hospital Medical Research Institute, Mumbai

Objectives: To pursue method verification as the first step tion tests were done [with particular reference to Thyroid in establishing standardization and harmonization of thyroid Stimulating Hormone (TSH)] by verifying reference intervals, function test, in order to eliminate error in reports. analytical accuracy and precision, inter-assay and intra-assay variations, analytical sensitivity, limit of detection, linear- Design and Methods: Verification of all the thyroid func-

123 S48 Ind J Clin Biochem

ity and reportable range i.e., (i) Analytical measurement manufacturer’s claim of within – Laboratory Precision SD range (AMR) and Clinically reportable range (CRR) as well (σl) or verification value (vv) for all level of control sera. as sigma metrics. Sigma metric for normal euthyroid level control serum was 8.77. Intra- assay variation C.V. 1.36% and 2.01% for Results: Our obtained reference range from verification low and high abnormal level control sera. Lowest limit of study was 0.62 – 4.22 micro IU/L. It showed high degree of detection was <0.01. AMR was also verified with C.V. 1.7%, analytical accuracy over AMR between two instruments (r2 1.85% and 2.51%, for low, normal & high level of control = 0.985 with linear regression equation of y = 0.94x + 0.11). sera, respectively. The assay was linear with different dilu- Precision was comparable with the manufacturer’s claim. tions. Lean concept (pneumatic system) was also verified Our (user’s) estimated within – run precision standard with high recovery percentage. deviation (SD) (sr) was lower than manufacturer’s claim of within – run precision SD (σr) or verification value (vv) for Conclusions: Verification protocol in the laboratory ensures all level of control sera. Similarly, our (user’s) estimated that accurate and precise results are reported. within – laboratory precision SD (sl) was also lower than

ANGIOTENSIN CONVERTING ENZYME ACE GENE POLYMORPHISM INSERTION/DELETION IN NORTH INDIAN INFERTILE MEN

M. Kaleem Ahmad*, A.A. Mahdi*, S. R. Hussain*, H. Naqvi*, S.P. Jaiswar$, S.N. Shankhwar# *Molecular Cell Biology Lab., Department of Biochemistry, King George’s Medical University, Lucknow, Uttar Pradesh, India. Departments of $Obstetrics & Gynaecology and #Urology, King George’s Medical University, Lucknow

Background: ACE is a membrane bound dipeptidyl carboxy (OAT), respectively. DNA of 221 infertile cases and 319 peptidase that generates vasoconstricting angiotensin II control subjects was analyzed for the insertion/deletion and inactivates vasodilating bradykinin. The ace gene en- polymorphisms of ACE using PCR amplification. codes two isozymes (sACE) and is found in many tissues Results: The ACE gene II, ID, DD genotype frequencies including vascular endothelial cells, whereas testis specific were obtained in 80.54%, 15.75%, 3.16% in cases and 81.19%, isozyme (tACE) is expressed exclusively in developing 15.98%, 2.82% in healthy controls. ACE D/D genotype was spermatids and mature sperms. We assessed seminal as- significantly more frequent in cases as compared to con- sociations of the ACE gene insertion/deletion polymorphism trols. in infertile men. Conclusions: ACE gene deletion polymorphism is associated Materials and Methods: A total of 540 men were investi- with abnormal seminal parameters, such that carriers of gated, divided into healthy fertile men, and infertile men the ACE deletion/deletion genotype might be a genetic with azoospermia, asthenozoospermia, asthenoterato- risk factor for male infertility. zoospermia Oligo zoospermia, Oligo-astheno zoospermia Oligo-terato zoospermia and oligoasthenoteratozoospermia IODINE DEFICIENCY ITS ADVERSE EFFECT ON SEXUAL AND REPRODUCTIVE HEALTH

MS Banerjee Prof. Emeritus, Acad. Medicine, MGM Medical College, Kishanganj Iodine deficiency in water and soil has a geographical tial component of thyroid hormones, necessary for body distribution common in Central Africa, Papua New Guinea, metabolism. The consequences of low thyroid function Andes Region of South America and Himalayan Valley. are impaired physical and sexual development, moreso in 300 million people live in these areas. Iodine deficiency girls, and poor reproductive performance and pregnancy is also prevalent in some developed countries. Survey has outcome. Congenital hypothyroidism is the commonest shown lower iodine level in teenage girls in UK, and also cause of mental retardation with or without other features amongst pregnant mothers in Brussels. Iodine is an essen- of goitre. Iodine deficiency in early pregnancy by itself has

123 Ind J Clin Biochem S49

adverse effect on brain development of the embryo with assay of thyroid function profile. Results corroborate consequent irreversiblecentral nervous system disturbances published data. including mental retardation. Conclusion: The catastrophic effect if iodine deficiency and Materials and Methods: Teenaged girls, prospective and the hypothyroidism are preventable by consumption of expectant mothers in Ethiopia, North Africa and those iodized salt and use of iodized oil as vaccine. around Calcutta. Study included Clinical Evaluation and

PATIENTS SAFETY AND CLINICAL BIOCHEMISTRY LABORATORY

Kaushal K. Srivastava Affiliate, Indian Confederation for Healthcare Accreditation

The healthcare system is a highly complex operation. De- The diagnostic errors have been frequently underestimated spite best efforts of the healthcare personnel, things keep in the clinical practice. A Biochemical Laboratory plays an on going wrong. These produce unintentional harm to the important role in clinical diagnosis. A laboratory error is patients. As such, patient safety must be considered as any defect occurring at any part of the laboratory cycle, one of the leading healthcare challenges. from ordering tests to reporting, interpreting, and reacting to results. Although they have been traditionally identified A medical error is traditionally defined as an unintended with analytical problems and uncertainty of measurements, act, the failure of a planned action to be completed as the vast majority of these arise from the extra-analytical intended, the use of a wrong plan to achieve an aim. Medi- activities of the total testing process. Pre analytical er- cal errors can be classified according to several models, rors are the first cause of variability in laboratory testing such as the clinical pathway (i.e., diagnostic, treatment, followed with subsequent processing through entire turn prevention and others), the resulting harm to the patient over time. (i.e., near misses, no harm or harmful incident). Medical errors can also be classified in skill-based slips and lapses The audit of system errors is seldom carried out in a clinical (i.e., errors of action), or rule and knowledge-based mis- laboratory set up. Such data are not available from Indian takes (i.e., errors of intention). Most errors result from the Laboratories and only a few from other developed health combination of active failures and latent conditions. care systems. This challenge is required to be taken up in the interest of Patients, the first and the last consumer in the health care industry.

123 S50 Ind J Clin Biochem

SYMPOSIA XI : FREE PAPER

IMMUNOFLUORESCENCE STUDY ANALYSIS OF 450 POSTIVE CASES OF VARIOUS AUTOIMMUNE DISEASES AT A TERTIARY CARE HOSPITAL AND RESEARCH CENTRE

Jagathlal Palliparambil Chandrasekharan; Lekshmi Krishnan Nair Geethakumari

Aim: To study about the percentage of incidences falling inflammatory arthritis, SLE, Scleroderma, Paliandromic in each autoimmune diseases and their correlation with rheumatism, poly myalgia rheumatica, and poly arthral- CRP (c reactive protein) and ESR (erythrocyte sedimenta- gia. tion rate) values. • Among this, Rheumatoid arthritis leads first with 29.98%(122 numbers), SLE comes second with 22.26% Objectives: As Amrita Institute of Medical Sciences and (91 numbers) Research Centre, Kochi, Kerala is a tertiary care hospital • Of the total 450 cases, rheumatoid arthritis and and academic, research institute, the population of patients other ostoe arthro inflammatory disorders constitute who get benefits from this hospital, represents the whole 49.22%. population of Kerala. So present study illustrates the au- • Rheumatoid arthritis is having a mean value of 27.29 toimmune disease prevalence in the state of Kerala. and standard deviation of 33.90 for C reactive protein. Materials and Methods: A median of 50 with minimum 10 and maximum 90 for • The study includes retrospective analysis of 450 positive ESR. cases of various autoimmune diseases diagnosed with • Regarding SLE, mean value of 17.98 and standard de- immunofluorescence analysis. viation 29.41 for CRP. A median of 50 with minimum • We got 5 immunofluorescence patterns which remark of 2 and maximum of 93 for ESR the positive cases. They were Homogenous, speckled, Disscusion: Centromeric, Nucleolar, and Mixed patterns. • The autoimmune patients are subdivided into 2 groups As IF pattern study gives a direct visual impression of depending upon the diagnosis 1) organ specific and 2) ANA. The diagnosis accuracy is more reliable than other systemic autoimmune disorders investigative method like ELISA. • Out of 450 autoimmune cases, 335 numbers of cases In kerala female to male ratio of auto immune disorders analysed for correlation with CRP and 402 numbers of are 7.7: 1 cases analyzed for correlation with ESR. Rheumatoid arthritis is the predominant auto immune dis- Results: order, with other osteo arthro inflammatory disorders. • Out of 450, positive cases 53.1% are belonging to speckled, 36.2% are homogenous, 5.6% are nucleolar, CRP mean value of SLE is less than that of Rheumatoid 4 % are centromeric and 1.1% is mixed patterns. arthritis. • The study include 391 females and 51 males Blood viscosity comparatively less in rheumatoid disorders • Out of 450 autoimmune patients 407 (90.44%) belonging than SLE to systemic autoimmune diseases and 43 (9.5%) belong- ing to organ specific autoimmune diseases group Conclusion: This study points out more detailed research • Systemic auto immune diseases include Sjogren’s syn- works required to sort out genetic, bio morphologic and drome, Rheumatoid arthritis, Auto immune haemolytic ecological back ground of kerala population for their day anemia, auto immune thrombocytopenic purpura, Fibro by day increasing prevalence of rheumatoid and other myalgia syndrome, inflammatory poly arthritis, juvenile osteo artho inflammatory auto immune disorders.

123 Ind J Clin Biochem S51

IMPORTANCE OF MIXED ACIDBASE DISTURBANCES IN CLINICAL PRACTICE

Dr. Kannan Vaidyanathan Clinical Associate Professor, Department of Biochemistry & In Charge, Metabolic Disorders Laboratory & ABG Facility, Amrita Institute of Medical Science, AIMS Ponekkara PO, Kochi, Kerala

Acid-base disturbances are usually classified into acidosis alkalosis in patients with COPD. Respiratory alkalosis may and alkalosis, which may be either of metabolic or respira- be associated with metabolic acidosis in salicylate poison- tory origin. However in actual clinical scenario, especially in ing, acute cardiopulmonary arrest and in Gram negative acutely ill patients, we see a mixture of different acid-base septicemia. Metabolic acidosis and respiratory acidosis are disturbances. Mixed acid base disturbance is defined as seen in cardiopulmonary arrest and respiratory failure with simultaneous presence of two or more acid-base disorders. anoxia. Respiratory alkalosis and metabolic alkalosis are Any kind of mixed disturbance is clinically possible, except seen in congestive cardiac failure, diuretic therapy, hepatic respiratory acidosis and respiratory alkalosis, in a given failure and pneumonia. patient at the same time. Recognition of mixed acid-base The commonest type of triple disorders is respiratory disturbance is very important because treatment plans will acidosis or respiratory alkalosis existing along with meta- depend upon the actual disturbance (s) present. bolic alkalosis and metabolic acidosis and may be seen in Suspicion of mixed acid base disturbance should be seri- cardiopulmonary resuscitation, COPD, advanced congestive ously considered when – heart failure. Patients with end stage renal disease are also 1. Expected compensatory changes are not seen, prone to complicated forms of mixed disturbances. 2. Changes in pCO2 and [HCO3-] are not as anticipated for It must be emphasized that the diagnosis of mixed acid- a simple disorder, especially with a normal pH, and base disturbances in not always easy. In this presentation, 3. Delta ratio is abnormal. we shall discuss some clinical scenarios where mixed acid- Chronic respiratory acidosis may co-exist with metabolic base disturbances were seen.

EFFECT OF YOGA AND NATUROPATHY ON IL 6 LEVELS IN PATIENTS SUFFERING FROM RHEUMATOID ARTHRITIS

Dr Ranjna Chawla, Dr Bhawna Singh, Dr VK Gupta, Dr Gaurav Thakur*, Dr Manjunath RM*, Aparna, Dr Rukmani Nair* Department of Biochemistry, GB Pant Hospital, *Department of research, Bapu Nature Cure Hospital and Yogashram, New Delhi

Introduction: Pathogenesis of rheumatoid arthritis (RA), an IL6 levels more than 6 pg/ml were selected and randomly autoimmune inflammatory disease, involves inflammatory divided into two groups– Gp1(n=10): took only allopathic cytokines like interleukin-6 which is found in abundance treatment; Gp2 (n=10): practiced yoga (pranayam and aasans) in the synovial fluid and serum of patients with RA and and naturopathy (hydrotherapy and message therapy) along level correlates with disease activity. Present study was with allopathic treatment. IL-6 levels were estimated in both aimed to see effect of yoga and naturopathy, that has groups after respective treatments for six months using gained tremendous magnitude in recent times, on IL-6 commercially available ELISA kits (Dichrome). levels in patients of RA. Results: Mean of IL-6 levels in patients (n=20), of RA at Materials and Methods: Study was conducted in department the time of diagnosis was 10.5± 6.5 pg/ml. IL-6 levels in of Biochemistry, GB Pant Hospital, in collaboration with patients belonging to group 1 were significantly lower department of research, Bapu Nature Cure Hospital and Yo- (2.4±1.6 pg/ml), (p value < 0.05) as compared to group gashram. Blood samples were collected from 20, radiologically 2 (6.1±4.8 pg/ml) after six months of their respective and serologically proven RA patients. Patients (n=20) with treatments.

123 S52 Ind J Clin Biochem

Conclusion: Significant reduction in levels of IL-6 in patients to allopathic cure emphasizes importance of natural healing who underwent regular yoga and naturopathy in addition measures in order to limit inflammation in RA.

FATTY ACID AS BIOLOGICAL MODULATORS OF TRANSCRIPTION FACTORS REGULATING FATTY ACID METABOLISM

Ransi Ann Abraham1, Lakshmy Ramakrishnan1, Rajinder Parshad2, Seenu V2, Prabhakaran D3, Bahl V K4 1Department of Cardiac Biochemistry, All India Institute of Medical Sciences, Delhi 2Department of surgery, All India institute of Medical Sciences, Delhi 3Centre of Chronic Disease Control, Delhi 4Department of Cardiology, All India Institute of Medical Sciences, Delhi

Objective: Fatty acids are unique macromolecules as they examined by the nonparametric Spearman’s rank correla- act as biological modulators of transcription factors and tion test. regulate their own metabolism by controlling the activ- Results: The expression of PPARγ and SREBP1c was signifi- ity or abundance of transcription factors of fatty acid cantly correlated (r=0.4 p<0.005). The correlation of 0.24 metabolism either by RNA processing and RNA stability. and 0.22 for linoleic acid and total PUFA (poly unsatu- PPAR-γ (Peroxisome Proliferator Activated Receptor) and rated fatty acid) was observed with PPARγ. A significant SREBP-1c (Sterol Regulatory Element Binding Protein) are correlation was observed for adipose tissue level of DHA transcription factors expressed primarily in adipose tissue. and PPARγ expression (r=0.33 p<0.03) which remained We have studied the relation of fatty acid assessed in significant (r=0.87, p<0.0001) after adjusting for BMI, insulin adipose tissue with the transcription factors. and energy consumed. Methodology: Adipose tissue was collected from 50 healthy Conclusion: The beneficial effects of PUFA have been subjects undergoing elective abdominal surgery. Fatty acid attributed to their ability to act as modulators. PUFA was assessed in the tissue by gas chromatography. The modulate plasma lipids by their effect on transcription expression of PPARγ and SREBP-1c were studied by real factors that regulate their partition into lipid oxidation or time PCR. Significance of the correlations between the synthesis. High intakes of PUFA are known to upregulate fatty acid and the expression of transcription factors was SREBP-1c and inhibit SREBP-1c. An upregulation of PPARγ often leads to decreased levels of SREBP 1c.

NONINVASIVE HYDROGEN BREATH TEST FOR MEASUREMENT OF OROCECAL TRANSIT TIME AND SMALL INTESTINAL BACTERIAL OVERGROWTH IN INFLAMMATORY BOWEL DISEASE

S V Rana, S Sharma, J Kaur, KK Prasad, SK Sinha, K Singh Department of Super Specialty Gastroenterology, Post Graduate Institute of Medical Education & Research, Sector 12, Chandigarh

Background: Inflammatory bowel disease (IBD) is chronic SIBO in UC & CD indian patients. inflammatory disorder with unknown etiology. IBD consists Materials & Methods: 137 patients of IBD & 115 healthy con- of ulcerative colitis (UC) and Crohn’s disease (CD). These trols were enrolled in present study. OCTT was measured two conditions share common features- diarrhea, bloody by Lactulose hydrogen breath test. Time taken for rise stools, weight loss, abdominal pain, fever and fatigue. ≥10ppm over fasting value was taken as OCTT. All patients Small intestinal bacterial overgrowth (SIBO) and orocecal and controls were investigated for SIBO after ingesting 80 transit time (OCTT) has not yet been studied in CD and gm of glucose dissolved in 250 ml water. Analysis of end UC Indian patients. expiratory air was done at the unset of test & every 15 Objective: This study was planned to measure OCTT & minutes for 2 hours. Rise in hydrogen ≥12ppm over the

123 Ind J Clin Biochem S53

baseline value within two hours of ingesting glucose was colitis patients (122.1±51.8ppm). Percentage of SIBO (22.62%) taken suggestive of SIBO. was also significantly higher in IBD patients as compared to controls (0.86%) which was further higher in Crohn’s dis- Results: Mean±SD of OCTT in IBD patients (130.5±50.8ppm) ease (45.2 %) patients as compared to UC patients (17.8%). was significantly higher as compared to controls (90.5±30.6 Conclusion: This study indicates that increased SIBO in UC ppm). Furthermore, OCTT was significantly higher in Crohn’s & CD patients may be due to increased OCTT. disease patients (138.9±49.9ppm) as compared to ulcerative

INSULIN RESISTANCE AND MEAN PLATELET VOLUME: RELATIONSHIP BETWEEN NON ALCOHOLIC FATTY LIVER DISEASES AND ATHEROSCLEROSIS

Subir Kumar Das Department of Biochemistry, College of Medicine & JNM Hospital, WBUHS, Kalyani, Nadia

Non-alcoholic fatty liver disease (NAFLD) is strongly associ- NAFLD patients and compared with 77 normal subjects in ated with abdominal obesity, type II diabetes mellitus, and this study. Insulin resistance (IR), assessed by homeostasis dyslipidemia, and most patients have evidence of insulin model assessment index (HOMA-IR) was confirmed in 70% resistance (IR). IR, the pathogenic driver of the metabolic patients with NAFLD and 42% patients fulfilled minimum syndrome, refers to a constellation of features such as criteria for IRS. Body mass index and waist circumference overweight/obesity, glucose intolerance, dyslipidemia and levels were higher in the NAFLD group when compared hypertension, all of which are important risk factors for to the controls. There were no differences between the cardiovascular disease (CVD). Low-grade inflammation two groups regarding LDL cholesterol levels, whereas HDL is typically linked to NAFLD. Thus, NAFLD shares many cholesterol levels were lower in the NAFLD group. NAFLD features of the metabolic syndrome, a highly atherogenic patients showed elevated levels of pro-inflammatory cy- condition, and this has stimulated interest in the possible tokines (TNF-α1, IL-6) and TGF-β1. But, the results of this role of NAFLD in the development of atherosclerosis. Mean study do not show any difference in MPV levels between platelet volume (MPV), a determinant of platelet activation, subjects with NAFLD and controls. Thus we conclude is an emerging risk factor for atherosclerosis. We evalu- that MPV might not be involved in the mechanism(s) of ated the relationship between IR and MPV among 105 increased cardiovascular risk in NAFLD.

ROLE OF ALCOHOL IN CAUSING NEURONAL APOPTOSIS A DOSE DEPENDENT STUDY

Sukhes Mukherjee Department of Biochemistry, VCSG Govt Medical College& HNB Base Hospital, Srinagar, Uttarakhand

Ethanol has a negative impact on human health. Moder- during ethanol metabolism and induction of pro-apoptotic, ate to heavy doses of alcohol have deleterious effects anti-apoptotic or other pathways. Ethanol intake dose and especially on brain and its different compartments. dependentlyupregulated COX-2 expression, iNOS expres- Excessive alcohol intake often results in life-threatening sion, p38 MAPK pathways and also cell death in brain medical disorders stemming from selective cell and tissue (cerebrum, thalamic area, mid brain hippocampal region injury and ultimately cell death. Oxidant stress caused by and cerebellum), which was evidenced by Caspase-3 and alcohol metabolism is a common element in cellular injury Caspase-8 assays. These findings indicated that exposure in brain. In the present study, we investigated the role of to higher concentration of ethanol induced brain dam- natural antioxidants like grape polyphenols and Vit E in age triggering the inflammatory processes and leads to combating the ROS balance in alcoholism for a dose of neuronal apoptosis by TUNEL assay. Here in this study 4g /Kg b.w./day of ethanol for a period of 4 months in we aimed in highlighting the role of natural antioxidant male Wistar rats. Enhancement of tissue injury with ethanol i.e. grape skin and flesh as a protective effect in alcohol consumption is operated by inflammatory mediators and induced brain damage. their modifications, generation of reactive oxygen species

123 S54 Ind J Clin Biochem

SYMPOSIA XII : FREE PAPER

ASSESSMENT OF SERUM MINERALS AND CLIMACTERIC SYMPTOMS IN PERIMENOPAUSAL WOMEN BEFORE AND AFTER YOGA THERAPY, A PRELIMINARY STUDY

1Alok Kumar, 2Archana E., 3Akshatha, 4Mrs. Gayathry Nayak, 5Dr. Revathi P. Shenoy, 3Dr. Anjali Rao 1Dept. of Biochemistry, ICARE Institute of Medical Sciences & Research, Haldia; 2Dept. Of Biochemistry, Manya Institute of Medical Sciences, Mandya; 3Dept. Of Biochemistry, Maleka Manipal Medical College, Manipal; 4Dept. Of Yoga Therapy, Kasturba Medical College, Manipal. 5Dept. Of Biochemistry, Kasturba Medical College, Manipal, India

Background: Many women report an increased level of were included in the study. Yoga intervention was given anxiety, irritability and mood swings during their perimeno- on a daily basis (45 minutes duration) for 12 weeks. The pausal period. Studies show that physically active people climacteric symptoms were assessed by Greene’s Climac- can reduce their anxiety and depression and yoga practice teric Scale and biochemical parameters were analysed is the one such activity. Magnesium plays an important spectrophotometrically. role in body temperature regulation and heart palpitation Results: Climacteric symptoms were improved significantly associated with hot flashes during perimenopausal period. (p<0.001) after yoga intervention. Changes in serum cal- Calcium deficiency in the body causes restlessness and cium, magnesium, copper and ceruloplasmin were within awakefulness observed in perimenopausal women and normal physiological range. serum calcium is also associated with blood pressure and metabolic risk factor for cardiovascular diseases. Conclusion: The overall changes observed in the min- eral status and climacteric symptoms suggest that yoga Objective: To assess the serum mineral status and climac- therapy protocol can be effectively used to improve the teric symptoms in perimenopausal women before and after quality of life in perimenopausal women. Sample size of yoga intervention. our study was less, so further study with larger sample Methods: It is a single group, prospective study. Thirty size can be done. female subjects belonging to the coastal belt of Udupi Dis- Keywords: Perimenopause; yoga therapy; serum minerals; trict, Karnataka with perimenopausal symptoms and ability climacteric symptoms. to perform yoga, aged between 40-60 years (49.43±6.15)

SERUM AMINOTRANSFERASES ACTIVITY IN EPILEPSY AND BIPOLAR DISORDER: RESPONSE TO TREATMENT WITH SODIUM VALPROATE AND CORRELATIONS WITH CLINICAL PARAMETERS

Hariom Sharma, Dharmesh Gamit, Hardik Javia Department of Biochemistry, Govt. Medical College, Bhavnagar, Gujarat Aminotransferases are an accepted marker for Liver injury. of Bipolar Disorder and Epilepsy after administration of In this study, we investigated serum Aminotransferase Sodium Valproate and elevation was highly significant. levels in Epilepsy and Bipolar Disorder treated with So- (ALT:- 27±10 vs. 58±22 IU/L, p<0.001 and AST:- 29±9 vs. dium Valproate. 75 patients with Bipolar disorder and 75 63±31IU/L, p<0.001). After com¬pletion of the 3 month patients with Epilepsy were enrolled in the study. Patients treatment, aminotransferases levels in Epilepsy and Bipolar included in the study then subjected to various biochemi- Disorder pa¬tients significantly increased (10.47±4.54 nmol/ cal investigations and their ultrasonography of liver was mL/h, P <0.001). Our results suggested a need for monitor- also performed initially and after 3 months of treatment ing serum Aminotransferases levels in patients receiving with Sodium Valproate. Aminotransferase levels were Sodium Valproate. measured by IFCC recommended UV kinetic method. The Keywords: Aminotransferases, Epilepsy, Sodium Valproate, levels of aminotransferases enzymes increased in Patients Bipolar Disorder.

123 Ind J Clin Biochem S55

ASSESSMENT OF SERUM COPPER IN CLINICAL DEPRESSION

Dr Ipsita Mazumdar1, Dr Chittaranjan Maity2 1Asst. Professor, Dept of Biochemistry, 2Professor & Head, Dept of Biochemistry, KPC Medical College & Hospital, Kolkata

Background: Copper, an essential trace element, is in- Materials & Methods: Copper was analysed on Microlab volved in a large number of bodily metabolic processes, 300 Semi-autoanalyser (Merck) employing Direct Colorim- including being co-factor in enzymatic activities related etry method with Spinreact Copper kit (Color 3,5-DiBr- to brain neurotransmitter function. It is a component of PAESA), in serum samples of 49 patients (28 female, 21 several metalloenzymes linked to dopamine synthesis, male; 25-65yrs) suffering from depression and on 45 (25 involving either antagonism of dopamine production or female, 20 male) age matched controls. catalysis of its breakdown. As dopamine is implicated in Results: The level of copper was found to be significantly clinical depression, copper homeostasis may be particularly (P<0.05) higher in patients (124.71+28.6) compared to relevant in its etiology, as an excess of copper may be controls (112.96+20.1). associated with dopamine dys-regulation. Various studies have indicated that excessive copper and zinc levels in Conclusion: Our study indicates that serum copper level blood may cause brain dysfunction, including depression, is generally higher in people suffering from depression schizophrenia, panic disorder etc. than in normal individuals. Hence we conclude that serum copper assay is a cost-effective, simple & readily available Aim: Considering this background, the present study was investigation that helps in the identification & diagnostic conducted to assess serum copper levels in patients suf- approach of clinical depression. fering from clinical depression & to determine any abnor- mality in its levels.

MEASURING THE EFFECT OF DIFFERENT PARAMETERS ON THE CATALYTIC ACTIVITY OF IMMOBILIZED ΒGALATOSIDASE FROM YEAST

Sandhya Hora1, Manoj Gupta2, Mamta Gupta3 1Research Assistant, Central Research Laboratory, Department of Biochemistry, Shri Ram Murti Samarak-Institute of Medical Sciences, Bareilly, Uttar Pradesh; 2,3PG Students, Department of Biochemistry, Shri Ram Murti Samarak-Institute of Medical Sciences, Bareilly, Uttar Pradesh.

Background: Enzyme catalysis is intermediate between plyacrylamide beads and alginate beads were used to homogeneous and heterogeneous catalysis. In general, analyze the catalytic activity of β-galatosidase on differ- it is similar to inorganic heterogeneous catalysis and ent parameters. sometimes it is called micro heterogeneous catalysis. Ef- Results: The effect of temperature, pH, and amino acids ficient enzyme catalysis depends on high specific nature on normal β-galatosidase and on immobilized β-galatosidase of enzyme, pH, temperature, optimum temperature, col- were analyzed. Immobilized enzyme showed the decreasing loidal nature, activator or co-enzyme, inhibitors or poisons. conversion level of lactose. Polyacrylamide beads showed Immobilized β-galatosidase show high activity towards decreasing conversion level of lactose at different concen- both pure lactose and lactose in skim milk, and a better tration of lactose and same concentration of beads. Effect thermal stability also. of temperature on β-galactosidase was maximum at 400C Methods: β-galatosidase was extracted by Kluyveromyces for growth. pH profile (6.2 to 7.4) of β-galactosidase was lactis and enzyme assay done by GOD/POD method. Partial studied at 280C & 370C. Highest lactose conversion (4.44%) purification of enzyme was done by precipitation with am- was noted at pH-6.8. monium sulphate, DEAE-cellulose column chromatography Conclusion: It was found that the β-galactosidase from followed by SDS-PAGE. Immobilization of K.lactis cells was K.lactis showed an optimum activity at temperature 400C, done by Entrapment method in which we agarose beads,

123 S56 Ind J Clin Biochem

pH 6.8. Permeabilization of the cell wall with chloroform able permeabilized cells give better result then viable increased the enzyme activity. The immobilized non vi- immobilized cells.

MULTIPLEXING OF BONE TURNOVER MARKERS

Meena Desai National Institute for Research in Reproductive Health (ICMR), J.M. Street, Parel, Mumbai

With an increasingly ageing population, osteoporosis and detection of multiple analytes, potentially higher sensitiv- its related fractures is fast becoming an important public ity, miniaturization of assay procedures, and reduction health issue. Biochemical markers of bone metabolism in sample and reagent volumes. Recently, beadbasedand play an important role in the assessment of bone diseases electro-chemiluminescencebased multiplex assays have such as osteoporosis, arthritisand bone metastasis with been developed andpromoted for being far more costef- cancers. Serum C-terminal telopeptide of type I collagen fective than traditional ELISA measurements. However, (CTX-I), N-terminal propeptide of type I collagen (PINP), the most commonly arrayed proteins are antibodies since Osteocalcin (OC) and Pyridinium crosslinks (PD/DPD) are they are robust molecules that can be easily handled and among the most sensitive bone turnover markers for immobilized by standard procedures on flat surfaces such evaluating osteoporosis. Currently, these biomarkersare as glass slides (chips) or membranes. The most widely measured individually by manual or automated immunoas- adopted strategy consists of a multiplex adaptation of the says. Enzyme-Linked Immuno-Sorbant Assay (ELISA), the classical antibody sandwich assay, where a pairof antibod- most widely used and validated method is limited by its ies binds to discrete recognition surfaces on each protein. ability to measure only a single protein in each sample, An antibody based array technology for bone markers is time consuming with sample volumes of 50 to 300 ul for currently under development for routine diagnostic ap- each assay. To overcome the limitations, theassay meth- plications. Both the ELISA and Multiplex measurement ods could bereplaced by multiplex microarrays. Multiplex technology in the current context will be reviewed. arrays have several advantages such as the simultaneous

VARIATION OF MAJOR CATIONS & NA+K+ATPASE IN ERYTHROCYTES & ITS RELATION BETWEEN THE SERUM LEVEL OF THESE CATIONS ALONG WITH GLUCOSE, UREA & CREATININE IN VEGETARIANS & NONVEGETARIANS A CROSS SECTIONAL STUDY

1Dr Smita S Sonoli, 2Mohit Adhikary, 2Dr Anuradha B Patil, 3Dr Vanishree B J, 3Basant Joshi, 4Dushyant Kumar 1,2,3Department of Biochemistry, J N Medical College, Nehrunagar, Belgaum, 4RMRC (ICMR), Nehrunagar, Belgaum

Abstract: Lacto-vegetarians, lacto-ovo-vegetarians & pesco- K+ATPase & the status of serum glucose, urea and crea- vegetarians are broadly put as vegetarians and who con- tinine in healthy vegetarians and non-vegetarians. sume animal products as meat, poultry, fish, seafood at Secondary Objective: Co-relation amongst the above men- least once a week are non-vegetarians. The diet is the tion parameters. key factor for tolerance to glucose, urea, salt retention & many enzymes. Na+-K+-ATPase, an enzyme present Type of Study: Cross-sectional. on the surface of RBC membrane, Na+& K+ are seen to Materials & Methods: The institutional board approved the alter with levels of urea and creatitnine, in the serum study & all volunteers gave written consent. 50 vegetarians and erythrocytes with an altered glucose level and blood and 50 non-vegetarians within the age group 30-40 years pressure. Hence this study was undertaken to analyse the were selected via the aid of a questionnaire & analysis above hypothesis of Na+, K+ and Na+-K+ATPase in serum & erythrocytes & Primary Objective: Estimatation of levels of serum and urea, creatinine & glucose in serum was done. Erythrocytes erythrocyte Na+, K+ in association with membrane Na+- isolated from samples were washed lysed and used for

123 Ind J Clin Biochem S57

estimation of sodium and potassium by flame photometer. Oxidase’s method, urea by DiAcetyl Monoxime’s (DAM) ATPase activity measured in by the method of Raccah and method and creatinine by Jaffe’s method. et al. Serum glucose is estimated by Glucose Oxidase- Per-

ESTIMATION OF PROTEINTHIOLS IN VERY LOW BIRTH WEIGHT NEONATES BEFORE AND AFTER RECEIVING TPN

Sumantha H.D.*, Mithun Nath*, Anjali Rao*, Kishnananda Prabhu*, Jayashree Purkayastha#, Soundaram V# *Department of Biochemistry &#Department of Pediatrics, Kasturba Medical College, Manipal

Introduction: Very-low-birth-weight infants are born at major systems that control the cellular redox potential & a time of otherwise rapid intrauterine brain & body prevent oxidative damage. This study is carried out with growth. These infants have limited endogenous energy an objective of estimating protein thiols in the plasma of stores. Specificneonatal morbidity (ventilator & oxygen neonates before and after receiving TPN. dependent infants) may also increase metabolic demands Materials and Methods: Hemodynamicallystable, low birth by as much as 25% compared to the controls. Rapid es- weight infants with a birth weight <1250 g are included tablishment of postnatal nutrition is essential to provide in the study and protein thiols in the plasma is measured continuous administration ofnutrition. Thus total parental using “Spectrophotometric method using DTNB-Ellman’s nutrition is provided to achieve a body composition & method”, on the day of birth as well as at the end of growth similar to those of a normal fetus of the same TPN. postmenstrual age. Result: No significant changes in the protein thiols are Objective: Total Parental Nutrition (TPN) is a potential observed so far. More significant results are expected in source of oxidants. A balance between oxidants & anti- future as the study is ongoing. oxidants is instrumental to maintain competent immune system. Thiol-disulfide interconversions play crucial role in Conclusion: The oxidative stress in the VLBW infants is not the chemistry of biological systems. They participate in significantly altered before and after receiving TPN.

123 S58 Ind J Clin Biochem

POSTERS

AUTOMATED HPLC SCREENING OF NEWBORNS FOR SICKLE CELL DISEASE

Dr. Mrs. Leela. G. Abichandani, Dr. Kanchan K. Sonone Department of Biochemistry, Grant Govt Medical College and Sir J.J. Group of Hospitals, Mumbai, Maharashtra, India.

Abstract: Sickle cell disease is a significant global public Results: We found that, birth prevalence of sickle cell trait health issue. Complications of this disease are numerous. was two out of hundred births ie; 2% among the total Newborn screening helps to diagnose the disease even population and 8% among backward and tribal communi- before the development of sign and symptoms of the ties of our society. disease. This would enable early detection and therefore Conclusion: 8% (as in our study) sickle cell trait prevalence early management that can improve both morbidity and among backward and tribal communities should be taken mortality, since it is a congenital genetic disorder. Symp- into consideration for recommending a universal neonatal toms usually don’t occur before 4 months of age. screening programme in the rural and hilly areas of Maha- Objective: To find the birth prevalence of sickle cell anae- rashtra and selective implementation of screening among mia and sickle cell trait among the hundred full term newborns in the cities. Thus, Neonatal screening is highly newborns screened at J.J. Hospital over a period of two significant, to identify babies with sickle cell disease and months. commence comprehensive care.

Technique: Umbilical cord blood (100 samples) of the new- Keywords: Screening, Sickle cell disease, HPLC. born was taken and thus was left unscratched. Automated HPLC was chosen for this purpose.

SCREENING OF GLUCOSE6PHOSPHATE DEHYDROGENASE DEFICIENCY IN NEWBORN BABIES

Dr. L.G. Abichandani; Dr. Raviraj Naik Grant Government Medical College, Mumbai

Abstract: The early characterization of G6PD activity Results: At the end of the study the prevalence of glucose- provides an aetiological diagnosis for neonatal jaundice, 6-phosphate dehydrogenase deficiency were found to be as well as the opportunity to give the newborns family 1 % and another 2 % of the samples were found to be information concerning the prevention of complications G6PD deficiency carriers. like hemolytic crisis and jaundice and mortality associated Considering its high incidence in our country, a neonatal with G6PD deficiency. screening programme for G6PD deficiency should therefore Material & Method: A total of 100 blood samples were be taken into account in national health schemes. collected from umbilical cord of fullterm newborn babies Keywords: screening, G6PD deficiency, qualitative dye in labour room of Obstetrics and Gynaecology depart- decolorization method. ment and analyzed at Biochemistry laboratory of Grant Government Medical College; Mumbai. All the samples were analyzed using a Motulsky qualitative dye decolor- ization method.

123 Ind J Clin Biochem S59

COMPARATIVE STUDY OF SERUM ELECTROLYTES BY FLAME PHOTOMETER AND AUTOANALYZER

Dr. Leela G. Abichandani, Dr. Shrikant C. Raut Department of Biochemistry, Grant Government Medical College, Mumbai

Abstract: Electrolytes are present in the human body, and was faster and with less quantity of serum sample as the balance of the electrolytes in our body is essential compared to Sherwood Scientific Model 420 flame pho- for normal functioning of our body. Common electrolytes tometer which requires diluted serum sample, which may important for quantitative estimation includes sodium and introduce manual error. Fully Automated Olympus AU400 potassium. In 1930’s, chemical methods were used to mea- autoanalyzer measures serum electrolytes with ion-selective sure serum electrolytes. To overcome disadvantages like electrodes (ISE) which depends on the ionic activity, time consuming process, little accuracy, flame photometer whereas flame photometer measures the stoichiometric was devised and after three decades, autoanalyzer was concentration. Olympus AU400 autoanalyzer for electrolyte introduced. estimation is a closed system, using the calibrator of the same company, whereas Sherwood Scientific Model 420 Objective: Measurement of serum electrolytes by means flame photometer is an open system. Olympus AU400 of Sherwood Scientific model 420 flame photometer were autoanalyzer can simultaneously determine glucose, urea, compared with measurements made by Fully Automated protein, etc. alongwith electrolytes. Olympus AU400 autoanalyzer to determine the efficiency of instrument. Conclusion: Determination of serum electrolytes by Olym- pus AU400 autoanalyzer was more efficient, sensitive Results: Determination of serum electrolytes showed slight and accurate than Sherwood Scientific Model 420 flame difference in the results. Measurement of serum electro- photometer. lytes by Fully Automated Olympus AU400 autoanalyzer

STUDY OF RENAL AND HEPATIC FUNCTION IN NEONATAL ASPHYXIA

Prachi Paliwal, Dr. Mrs. Meena Varma, Dr. M. K.S. Shaikh SAIMS, Indore, M.P.

Objectives: Birth asphyxia occurs when a baby does not 2. Serum Creatinine was estimated by JAFFE’S KINETIC receive enough oxygen before, during or just after birth. method. Birth asphyxia is a multisystem disorder. This study was 3. Serum SGOT and SGPT were estimated by MODIFIED undertaken to assess the hepatic function and renal func- IFCC method. tion in asphyxiated neonates and to correlate with severity Results: Blood urea and serum creatinine were significantly of birth asphyxia. High blood urea and serum creatinine higher in asphyxiated babies compared to the control values are associated with renal failure. Rise in hepatic group. (p<0.001). Blood Urea nitrogen/Creatinine ratio enzymes (SGOT, SGPT) indicates liver cell dysfunction. was significantly higher in asphyxiated babies compared Values of serum creatinine, urea, SGOT and SGPT were to control groups (p<0.001) on day 3 but insignificant on measured on 1st and 3rd day of life. day 1. The mean SGOT and SGPT of asphyxiated babies Methods: The present study included 50 asphyxiated and were significantly higher in asphyxiated babies compared 50 healthy neonates. Biochemical parameters such as blood to control groups (p<0.001). The rise also showed a sig- urea, serum creatinine, hepatic enzymes SGOT and SGPT nificant correlation with severity of birth asphyxia. were assessed for renal functions and hepatic functions Conclusion: We conclude that renal failure is a significant respectively. problem in asphyxiated neonates. A high level of urea and 1. Serum Urea was estimated by UREASE GLDH meth- creatinine on 3rd day of study is associated with renal od.

123 S60 Ind J Clin Biochem

failure in asphyxia. The rise in Serum hepatic enzymes is as well as to assess severity of birth asphyxia. Thus early indicative of liver cell dysfunction either due to hepato- diagnosis and treatment can be provided on the basis cyte necrosis or due to changes in cell permeability. The of renal and liver function tests particularly whose birth estimation of hepatic enzymes can be used for diagnosis details are not well recorded.

STUDY OF TRACE ELEMENTS IN MALIGNANT AND NONMALIGNANT ASCITES

Md Khaleel Pasha, M. Gopal Reddy, Qamar Ayesha Department of Biochemistry / Gastroenterology, Osmania General Hospital, Afzalgunj, Hyderabad, A.P. Introduction & Objectives: The diagnosis of ascites is and non malignant ascites patients when compare to con- made by clinical, noninvasive and invasive methods. The trols (34.62, 35.91 vs 102.41 p<0.001, p<0.001 respectively). conventional methods are inadequate in the differential The zinc levels in ascitic fluid samples in malignant group diagnosis of ascites due to its multitude causes. In this were considerably high (20.92 vs 16.70) when compared study trace elements were studied to explore in the dif- to non malignant group (p < 0.05). Similar results were ferential diagnosis of malignant and nonmalignant ascites noted in the iron levels in both the groups (serum 37.22, and understanding their pathophysiology. 48.91 vs 127.68, p < 0.001 respectively). The magnesium levels in serum samples of non-malignant ascites patients Materials & Methods: In the present study 70 cirrhosis are comparable to that of controls (2.00 vs 2.05) but the of liver (53 males, 17 females, 22-75 yrs), 53 ovarian ma- levels in malignant ascites patients are high when compared lignancy patients (53 females, 19-84 yrs) and 60 controls to that of controls (2.86 vs 2.05). were used. Trace elements such as magnesium, copper, zinc and iron in serum and ascitic fluid were quantified Discussion & Conclusion: Since trace elements are cofac- using standard biochemical methods. tors to certain enzymes and functional proteins which can influence their activity. Their deficiency or toxic levels can Results: The mean serum and ascitic fluid concentrations causes altered fluid and electrolyte homeostasis. Assessing of copper were significantly higher in ovarian malignancy the relationship between trace element levels and conven- patients compared to cirrhosis of liver patients (161.69 vs tional tests would be useful in the differential diagnosis 105.82; 111.90 vs 29.20 p <0.05 and p < 0.01 respectively). and prognostic evaluation. The zinc level in serum samples were low in both malignant

ROLE OF OXIDATIVE STRESS IN AGGRAVATING KIDNEY DYSFUNCTION IN CORONARY ARTERY DISEASE PATIENTS

Ritu Sharma, Mridula Mahajan Department of Biochemistry, Government Medical College, Amritsar

Background & Objective: Acute cardiac decompensation of clinical symptoms, ECG changes and TMT test wherever results in activation of hemodynamic and neurohormonal required. All the patients were screened for cardiac profile factors that lead to an acute drop in GFR resulting in (serum Total lipid profile, plasma SGOT, SGPT), blood glu- acute kidney injury. This relationship is referred to as cose and serum uric acid levels along with blood urea and Cardiorenal syndrome type 1(CRS), which usually goes serum creatinine to evaluate their kidney functions. Written unnoticed. The present study was designed to evaluate informed consent was obtained from all the patients as the occurrence of kidney dysfunction in coronary artery per The Ethical Committee of the Institute. disease (CAD) patients (CRS type 1) visiting the clinical Results: Ninety percent (90%) of CAD patients were ob- biochemistry laboratory. served to have renal disease which was revealed from Methods: A total of hundred (n=100) CAD patients visit- their significantly raised (p<0.001) blood urea and serum ing the clinical biochemistry laboratory were selected. The creatinine levels. Renal dysfunction was more pronounced diagnosis of CAD was done by the clinician on the basis in CAD patients suffering from congestive heart failure

123 Ind J Clin Biochem S61

along with hypertension. All these patients were advised Conclusions: Reduced free radical scavenging capacity of serum uric acid estimations by the clinician. A close look the body may be responsible for inflammatory conditions at serum uric acid levels interestingly showed that levels prevailing in the body in response to the injury to the cell were significantly low (p<0.01) in CAD patients having membrane causing organ dysfunction which could be the kidney dysfunction as compared to those without kidney involvement of kidneys in CAD patients. Hence it is very disease. Relatively low uric acid levels in these patients important to check the prooxidant-antioxidant balance indicate its quenching in scavenging of free radicals at very initial stages of a diseased condition involving owing to increased oxidative stress. The prevalence of inflammatory processes so that the advanced complica- increased oxidative stress was further supported from tions leading to organ dysfunction can be put to check significantly low activities of serum SOD and catalase in well in advance. these patients. Keywords: Cardio-renal syndrome, creatinine, urea, coronary artery disease, oxidative stress EVALUATION OF THERAPEUTIC ROLE OF FILARIAL ANTIGENS ON THE DEVELOPMENT OF TYPE 1 DIABETES

Manjusha Hivre, Nitin Amdare, Vishal Khatri, Ravi Yadav & M.V.R. Reddy Department of Biochemistry, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Maharashtra Background: Increasing evidence is available in support the blood glucose levels. Mice treated with rBmCys in of an inverse relationship between worm infection and T alum adjuvant showed significantly lower blood glucose helper type 1/17 (Th1/17) - based inflammatory disorder such level as compared to the diabetic mice treated with only as Type 1 diabetes suggesting the therapeutic potential of Alum (p<0.001). Also there was significant reduction in the helminth molecules in this condition. glucose level in diabetic group of mice treated separately with filarial native proteins (Bm mf’s and Bmmf’ES) com- Objective: To evaluate the therapeutic potential of filarial pared to diabetic mice treated with only Alum (p<0.002 immune modulators (Bm mf’s/ Bmmf’ES’/ rBmCys) on the andp<0.001). Almost 70% of the mice treated with Bmmf’ES development of type 1 diabetes. showed recovery from diabetes at the end of the experi- Methodology: A study was designed to validate therapeutic ment period. Although all mice had evidence of ongoing efficacy of filarial proteins (Bm mf’s/ Bmmf’ES’/ rBmCys) pancreatic islet cell inflammation by histology, micetreated in streptozotocin (STZ) induced diabetic mice. Following with filarial proteins had greater numbers of total intact experimental induction of diabetes the mice were either islets and non-infiltrated islets than untreated group of treated with or without the said proteins using alum as mice. adjuvant (25 μg) for 2 months. After treatment the blood Conclusion: These findings suggest that filarial derived glucose level and pancreatic histopathological changes proteins play pivotal role in the amelioration of disease were measured. condition in mice and act as novel therapeutic candidates Results: Therapeutic treatment of diabetic mice with filarial in the treatment of type 1 diabetes. proteins reduced the severity of disease by decreasing

ALPHA1 ANTITRYPSIN SERPIN LEVEL IN SOME LUNG DISEASES

Khushtar Anwar Salman,1 Sharique Ahmed,2 Humera Khan,3 & Akif Ehsan4 1,4J N Medical College, Department of Biochemistry Faculty of Medicine, Aligarh Muslim University, Aligarh 2Department of Allied Health Division, College of Health Sciences, Kingdom of Bahrain 3Department of Biochemistry, HIMS Lucknow

Human plasma contains a number of proteinase inhibitors control a number of functions such as fibrinolysis, coagu- which together form 10% of the total plasma proteins. They lation, complement activation, connective tissue turnover

123 S62 Ind J Clin Biochem

and inflammatory reactions. Alpha-1 antitrypsin (α1-ATT), in emphysema. Keeping this in view, (α1-ATT activity was a serine proteinase inhibitor (SERPIN) is found in highest measured in 60 patients (both males and females suf- concentration in plasma. α 1-ATT (anti-protease), normally fering from various lung diseases like, chronic bronchitis, inhibits elastase (protease) released during phagocytosis emphysema, bronchiectasis, asthma, pneumonia and tu- by neutrophils present in alveoli of lungs The protein berculosis. (age: 31- above 60 years) Among these 35% alpha-1 antitrypsin is a 52 kDa molecule produced primar- were smokers. ily in hepatocytes and released into the blood circulation Methods: α1-antitryptic activity was measured by the by the liver. The protein is present in all body tissues but method of Waheed & Salahuddin (1975) The level of serum appears to have its primary physiologic significance in the α 1-ATT activity in 25 healthy subjects served as control lungs, where it protects the healthy but fragile alveolar and was compared with the study group. tissue from proteolytic damage by enzymes like neutrophil elastase. Alpha-1 antitrypsin is an acute phase protein which Results: The present study establishes that the incidence means that production by the liver is subject to various of α 1-ATT deficiency was higher among male smokers. stimuli, including inflammatory mediators Therefore, protein Conclusion: Individuals with α 1-ATT deficiency have symp- levels in the circulation may vary depending on the medical toms suggestive of asthma and this can lead to misdi- condition of an individual. The normal serum concentration agnosis. Thus, to prevent the fatal outcome of α 1-ATT may range between 1.5 to 3.5 g/L (or 20 to. 48 μM) deficiency, an early diagnosis is recommended. Imbalance between the two enzymes, proteases and an- tiproteases causes the injury to lung tissue which results

EVALUATION OF IMMUNOMODULATORY POTENTIAL OF FILARIAL ANTIGENS ON THE DEVELOPMENT OF TYPE 1 DIABETES

Manjusha Hivre, Mohammad Sameer. M.H. Khan, Nitin Amdare, Ravi Yadav, Vishal Khatri & M.V.R. Reddy Department of Biochemistry, Mahatma Gandhi Institute of Medical Sciences, Sevagram, Maharashtra

Background: In recent years substantial epidemiological vals of 15 days) followed by the induction of diabetes and experimental evidence became available to support using streptozotocin (STZ). The mice was followed up by the hypothesis that infection with helminths can sup- monitoring their plasma fasting glucose levels. press the development of type 1 diabetes. Understanding Results: At the end of experiment period, the group of modulation of the host immune system by pathogens mice (n=7) pre-treated with rBmCysfollowed by induction offers rich protective approach. Hence, identification and of diabetes (rBmCys-STZ) showed significantly lower fasting characterization of helminths derived immunomodulatory blood glucose level compared to the levels in mice (n=7) molecules that contribute to the protective effect could pre-treated with only alum adjuvant (Alum - STZ) (p<0.01). lead to new protective approaches in type 1 diabetes. Also there was significant reduction in the blood glucose Objective: To evaluate the immunomodulatory and pro- levels in the mice pretreated withBm mf’s (n=7, p<0.001) phylactic effect of filarial immune modulators (Bm mf’s/ rBmmf’ES (n=7, p<0.001). Consistent with thesechanges Bmmf’ES’/ rBmCys) on the development of type 1 diabe- in glucose levels, mice pre-treated with filarial proteins tes. showed almost normal pancreatic islet cell architecture with minimal inflammatory change. Methodology: BALB/c mice (n=7-8) were pretreated with 4 doses of filarial derived proteins (Bm mf’s/ Bmmf’ES/ Conclusion: Present study provided evidence of exploiting rBmCys) in alum adjuvant (25μg) for 2 months (at inter- filarial derived proteins for the prevention against type 1 diabetesthrough immune modulation.

123 Ind J Clin Biochem S63

ASSOCIATION OF SERUM CALCIUM & PHOSPHORUS LEVELS WITH CHRONIC KIDNEY DISEASE CKD

Dr Milav H. Bhavsar, Dr Rinku R. Makadiya, Dr PranayJadav, Dr Rita M. Shah

Objective: To measure serum calcium & phosphorus levels 86% patients. Also among patients with or without CKD- in CKD patients &evaluate their association with CKD. low calcium levels were found in 85.7% patients with high phosphorus levels. Methodology: This is a descriptive analytical cross-sectional study carried out at Dhiraj Hospital, Piparia, Vadodara. Conclusion: Low calcium level & high phosphorus levels Total 71 indoor patients were enrolled in study provided are associated with patients with CKD also there is direct time period, 45 patients with known CKD &26 without correlation with high phosphorus & low calcium levels in CKD taken as control group. CKD or non CKD patients.

Results: In CKD patients low calcium levels were found Keywords: Chronic Kidney Disease (CKD), S. Phosphorus, in 80% patients & high phosphorus levels were found in S. Calcium.

EFFECT OF CHELATORS ON TYROSINASE ACTIVITY

Dr. S. J. Makhija and Dr. B.S. Badadhe Biochemistry Dept S.B.E.S. College of Science, Aurangabad Tyrosinase enzyme is a type of protein that helps to pro- It was observed that appart from inhibition; sodium azide duce chemical reactions in the body. Tyrosinase inhibitors could also act as activator in certain cases. All the chelators are important ingredients of someindications that help to showed maximum decrease in tyrosinase with beetroot as decrease some types of coloured spots on people’s skin. the sources of enzyme. We tried to study the effect of chelators like glycine, These studies may have a way to the pigmentation and cysteine and sodium azide on Tyrosinase isolated from depigmentation in skin diseases. different sources esp. potato. Other sources showed vari- able changes.

STUDY OF OXIDATIVE STRESS AND IRON OVERLOAD IN PATIENTS WITH VARIED ALCOHOL CONSUMPTION IN CRONIC ALCOHOLOLISM

Neelesh Deshpande, Manohar Muddeshwar Department of Biochemistry, Government Medical College, Nagpur

Oxidative stress has been increasingly implicated in the group had significantly lower serum SOD (P < 0.0001) pathogenesis and progression of cirrhosis. Chronic etha- GPx (0.0001) and significantly higher MDA levels and fer- nol administration is able to induce an oxidative stress. ritin levels (P<0.001) than those in control group. These AIM: The aim of this study is to demonstrate the effect results suggest that the decrease in serum SOD, GPx and of alcohol exposure on biomarkers of oxidative stress increase in MDA and ferritin levels are related to the Viz: Serum superoxide dismutase (SOD) and Glutathione alcohol consumption and status of liver cirrhosis clearly peroxidase (GPx), Malonilaldehyde (MDA) and ferritin in depicted with the combination of four biomarkers. Conclu- cirrhotic patients. Methods: All 50 decompensated cir- sions: The decrease in the antioxidant defense system and rhotic patients were divided into two groups with (28 the rising oxidative stress with leads to pathogenesis and Low alcoholic intake (ALC1), mean age: 42.7 ± 8.4 years; liver injury is suggested to be associated with iron overload 22 on high alcoholic consumption (ALC2), mean age: 42.2 and continued alcohol drinking behavior. Thus progression ± 7.9 years) with variable alcohol intake considering the in liver injury with continued drinking behavior underlines exposure period of 30 years. Results: The experimental importance of ceasation methods.

123 S64 Ind J Clin Biochem

A STUDY OF ASSOCIATION BETWEEN LIPID PROFILE AND WAIST TO HIP RATIOS IN DIABETIC PATIENTS OF WESTERN LUCKNOW

Seema Singh1, Kanchan Singh2 Vishnu Kumar2 and Ajanta Roy2 1Department of Physiology, Institute of medical Sciences, Integral University, 2Department of Biochemistry Era’s Lucknow Medical College and Hospital, Sarfaraz Ganj, Hardoi Road, Lucknow

Introduction: Indian population faces higher risk of diabetes automatic enzymatic analyzer (Erba Chem. 5 Plus, Transasia, and its complications. The waist to hip ratio is commonly Ltd., India). The waist to hip ratio is simply calculated by used as an indirect measure of lower and upper body fat dividing waist circumference by hip circumference as per distribution. Body mass index and waist circumference WHO norm of anthropometric measurement. independently contributed to the prediction of abdominal, Results: The results indicate that in females, statistically subcutaneous and visceral fat. In the present study, an negatively significant correlation (r = - 0.26) was found in attempt has been made to investigate the correlation of LDL- C in age group 41 – 50 years and in males of the waist to hip ratios with lipid profiles. same age group, positively significant correlations were Aim of study: To evaluate correlation between lipid profile noted in serum cholesterol (r = 0.48), LDL-C (r = 0.35) and waist to hip ratios in diabetic patients of Western and in Triglyceride (r = 0.35) and also in age group 61+ Lucknow. years positively significant correlations were found in males between waist to hip ratio and Triglyceride (r = 0.60). Material and Methods: This study was carried out on a total of 90 subjects in which 45 were male and 45 were Keywords: Serum cholesterol, Triglyceride, High Density female, attending OPD of General Medicine, Era’s Lucknow Lipoprotein – Cholesterol, Low Density Lipoprotein Cho- Medical College & Hospital, Sarfaraz Ganj, Hardoi Road, lesterol. Lucknow. Serum lipid profiles were determined with a semi-

STUDY OF SERUM TRANSAMINASES IN HYPOTHYROIDISM

Dr. K.S. Manjula1, Dr. Priyadarshini2 1Post-Graduate Student 2Associate Professor, Department of Biochemistry, Rajarajeshwari Medical College, Bangalore.

Background: Thyroid hormones regulate Basal Metabolic OPD of Rajarajeshwari Medical College & Hospital, Banga- Rate (BMR) and calorigenesis in tissues, including hepato- lore were taken as control. Estimation of T3, T4, TSH by cytes and thereby modulate hepatic function. In hypothy- ultrasensitive chemiluminescence, serum cholesterol, AST roidism cholesterol elevation occurs due to hypometbolism and serum ALT by enzyme kinetic method in Transasia EM which resuls in fatty liver causing elevation of transami- 360 autoanalyzer were carried out. nases in serum. Muscle involvement in these patients Results: 99% of the study group were females in the age also results in elevated AST. Raised serum transaminase group of 30-35 years. Serum AST (0-35 U/L) and serum activities in absence of any overt liver dysfunction can ALT (0-40U/L) were within normal reference range in both therefore be attributed to primary thyroid dysfunction. cases and controls. The aim of this study was to assess the impairment in liver function by estimating serum Aspartate Transaminase Conclusion: The likelihood of abnormal serum transami- (AST) and Alanine Transaminase (ALT) in patients with nases levels in hypothyroid patients is minimal. Early hypothyroidism. detection and better management of the thyroid diseases in recent times might be attributed as one of the factors Materials and Methods: 50 patients diagnosed with thy- for the same. roid disorders irrespective of duration of the disease were taken as cases and 50 healthy adults attending medical Keywords: Thyroid disorders, Transaminases.

123 Ind J Clin Biochem S65

POTENTIAL OF POLYHERBAL PREPARATION AGAINST GENTAMICIN INDUCED NEPHROTOXICITY IN RAT

Bharati D. Talele, Manojkumar Z. Chopda, Namrata V. Nemade and Raghunath T. Mahajan Department of Zoology, Moolji Jaitha College, Jalgaon

The present study was undertaken to evaluate the poly- used aminoglycoside antibiotic. In rats, renal damage is herbal formulation. Bark of Bahunia racemosa, pod of indicated by several folds increase of BUN (blood urea Tephrosia purpura, seed of Tectona grandis fruit of Tribulus nitrogen) and creatinine concomitant histological changes terrestris Seed of Dolichos biflorus and flower of Sphaer- in kidney tissue. In rat, damage is occurred around eight anthus indicus for its protective effect on nephrotoxicity days after gentamicin induced toxicity. The administration induced in Wistar rats by intraperitoneal administration of herbal mixture enhances the recovery rate of kidney of gentamicin 80 g/kg/d for eight days in rats. The study damage as indicated by decrease in BUN and creatinine. groups include six rats in each group. Administration of The polyherbal preparation showed very significant recov- herbal mixture at 250, 500 and 1000 mg/kg body weight ery on gentamicin induced nephrotoxicity. given by oral route was determined using serum creatinine, Key words: Gentamicin, Nephrotoxicity, polyherbal, Neph- serum albumin, blood urea nitrogen, and haematology as roprotective indicators of kidney damage. Gentamicin is an extensively

RELATION BETWEEN ANEMIA AND TYPE 2 DIABETES MELLITUS

Dr Rinku Makadiya1, Dr Milav Bhavsar1, Dr Pranay Jadav2, Dr Rita Shah3 1Resident, 3HOD & Prof, Department of Biochemistry, Smt. Bhikhiben K. Shah Medical Institute & Research Center, Piparia, Gujarat. 2Resident, Department of Community Medicine, Smt. Bhikhiben K. Shah Medical Institute & Research Center, Piparia, Gujarat

Objective: Estimation of Hemoglobin level in type 2 dia- Results: Anemia was 37% in patients with only diabetes, betes mellitus. 17% in diabetic patients with chronic kidney disease, 3% in patients with only chronic kidney disease. Anemia is Methodology: This is a descriptive analytical cross-sectional significantly higher in diabetes patient than chronic kidney study carried out in Dhiraj hospital, Piparia, Vadodara. diseaseor diabetes and CKD patient. A total no of 100 consecutive patients were enrolled; 25patients having Diabetes, next 25 patients having both Conclusion: Hemoglobin level is found in lowerlevel in dia- diabetes and chronic kidney disease, another 25patients betic patients than those patients who have only chronic having only chronic kidney disease (CKD) but no diabetes kidney disease or diabetes and chronic kidney disease. and another 25patients not having diabetes or chronic Keyword: Glycosylated hemoglobin (HbA1c), Hemoglobin, kidney diseasetaken as a control group. diabetes mellitus (DM), chronic kidney disease (CKD).

STRUCTURAL CHARACTERIZATION OF ADIPONUTRIN AND CHRONIC LIVER DISEASE

Atanu Kumar Dutta Demonstrator (Lecturer), Department of Biochemistry, IPGMER, Kolkata

Chronic liver disease is a leading cause of mortality, out what are the predisposing genetic factors in Indians morbidity and loss of productivity worldwide. With west- towards chronic liver disease which revealed Indians are ernization of lifestyle and dietary patterns the disease more likely to produce an inflammatory response in the burden is increasing further in India and Indian migrants liver compared to other ethnic groups. However till date in European countries. An attempt was made to find only one gene named Adiponutrin has been proved to be

123 S66 Ind J Clin Biochem

associated with higher incidence of chronic liver disease Joint Centre for Structural Genomics at Stanford Univer- of varied aetiologies. Isoleucin to Methionine substitution sity. A homology modelling analysis carried out through at 148th position of the protein increases the hepatic fat the ITASSER web based software and a network analysis content in Human. However the exact physiological role with STRING 9 software gave surprising insights into the of this protein is still not clear and knockout study in structure and function of this protein which is in addition mice failed to produce any pathological change in the to the known function. However computational design- liver. Hence it is customary to gain in depth knowledge ing of specific inhibitor to targets within this protein about this protein to modulate it for the betterment of would be absolutely necessary once the crystal structure the patients. A request for crystallographic analysis of the is known to find out if the predicted characteristics are protein was submitted through Nature Structural Biology indeed correct. Knowledgebase and is currently being carried out at the

HBA1C, HSCRP AND ANTHROPOMETRIC PARAMETERS EVALUATION IN THE PATIENTS OF DIABETES MELLITUS OF CENTRAL RURAL INDIA

Ajay Meshram, Udit Agrawal Professor and Head, Department of Biochemistry, J. N. Medical College (D.M.I.M.S.), Sawangi (M), Wardha

Association of hs-CRP in diabetic and non – diabetic sub- parameters were found to be high in diabetic subjects jects was studied in our rural based tertiary care hospital. compared with non-diabetic subjects. The high hs-CRP The analysis was done with 50 diabetic and 50 non-diabetic levels in diabetic subjects were also observed. individuals. Anthropometric and biochemical parameters Serum hs-CRP levels were positively related to anthropo- were studied to assess the association of hs-CRP within metric parameters. The relationship of hs-CRP with glycae- diabetes mellitus. mic control was studied with HbA1c, and it was positively hs-CRP, the golden marker of inflammation was analyzed correlated with hs-CRP. The results concluded that hs-CRP in diabetic and non- diabetic subjects. Anthropometric has strong association with diabetic individuals.

BIOCHEMICAL HYPERTHYROIDISM OF HYDATIFORM MOLE: “HOOKED” WITH A GRAPY CASE

Meghana. K. Padwal1, Gayatri Gawade2, Gaurav Karve3, R.R. Melinkeri4 1Associate Professor, 2Jr Resident II, 3Jr Resident III, 4Prof & Head, Department of Biochemistry; BVDUMC, Pune Background: Gestational trophoblastic disease (GTD) is a the measurement of Beta HCG by Chemiluminescence im- rare complication of pregnancy, ranging from molar preg- munoassay, we faced analytical challenges due to “High nancy to choriocarcinoma. Occasionally; GTD is complicated Dose Hook effect.” Patient also developed biochemical by hyperthyroidism, which may require treatment. This is hyperthyroidism but did not show any signs and symptoms thought to occur due to molecular mimicry between hu- of clinical hyperthyroidism during perioperative period. man chorionic gonadotrophin (HCG) and thyroid-stimulating Four months follow up after suction of mole showed hormone (TSH), resulting in cross-reactivity with the TSH decline in Beta HCG levels with normalisation of T3, T4 receptors. and TSH levels.

Case Report: A 20 year old female with 10weeks of preg- Conclusion: Quantitative rise in Beta HCG may not al- nancy, presented with severe abdominal pain, per vaginal ways reflect as clinical hyperthyroidism. However careful bleeding, vomiting and breathlessness. She was diagnosed monitoring of thyroid function is mandatory in all cases as a case of partial hydatiform mole. of molar pregnancies.

Results: We report highest levels of Beta HCG encountered Keywords: Biochemical Hyperthyroidism, High dose Hook in our set up in cases of trophoblastic diseases. During effect.

123 Ind J Clin Biochem S67

STUDY OF ANTIOXIDANT ACTIVITY OF CARYOTA URENS

Mrs. P.V. Ansulkar, Amruta Naik, Dr. Abhay Chowdhury Department of Biochemistry, Haine Institute, Parel, Mumbai

An antioxidant is a molecule capable of terminating the evaluated using two assays viz. DPPH Radical Scavenging chain reactions that damage cells by removing free radical Assay and Total Reducing Capacity. In DPPH assay, the IC50 intermediates and inhibit other oxidation reactions by re- values obtained for Hexane, DCM and Methanol extracts ducing stress responsible for many degenerative disorders. were 223.3μg/ml, 69.32μg/ml and 82.23μg/ml respectively. The palm caryota urenes Linn., amultipurpose tropical plant In the Total reducing capacity assay, activity increased in is believed to havemany medicinal properties. The purpose dse dependant manner for all the three plant extracts. of this study was to evaluate the in-virto antioxidant Keywords: Caryota urens, Antioxidant, DPPH assay, Total activity of the Hexane, DCM and Methanol extracts of reducing capacity. Caryota urens. The antioxidant activity of the extracts was

THE GLYCEMIC INDEX OF PARTIALLY REFINED YELLOW SUGAR AND PLANTATION WHITE SUGAR: A COMPARATIVE STUDY

Amruta Naik, Pallavi Ansulkar, Dr. Abhay Chowdhury Department of Biochemistry, Haine Institute, Parel, Mumbai

The Glycemic index (GI) is a measure of the effects of The main objective of this study was to estimate the com- carbohydrates in food on blood sugar levels. It estimates pare the GI values of the Partially Refined Yellow Sugar how much each gram of available carbohydrate (total and Plantation White Sugar. In-vivo study was conducted carbohydrate minus fiber) in a food raises a person’s by using Male wistar rats as a model and blood glucose blood glucose level following consumption of the food, analysis was done on semiautoanalyser by GOD-POD relative to consumption of glucose. Cane sugar requires method. In this study, it was observed that the GI value further processing to provide the free-flowing white table of the Partially Refined Yellow Sugar was 84% and GI value sugar required by the consumer. The completely refined of the Plantation White Sugar was 100%. From this we white sugar product is now over 99.9% sucrose and for all concluded that Partially Refined Yellow Sugar has low GI practical purposes contains no nutritional elements such value as compared to the Plantation White Sugar and thus as vitamins, minerals, proteins or fibers. Refined sugar is it can be used for the preparation of food items for the lethal when ingested by humans because it provides only diabetic patients and for the weight loss program. that which nutritionists describe as “empty” or “naked” Keywords: Glycemic Index, Partially Refined Yellow Sugar, calories. It lacks the natural minerals which are present Plantation White Sugar. in the sugar beet or cane.

HYPOGLYCEMIC AND ANTIHYPERLIPIDEMIC EFFECT OF AQUEOUS LEAVES EXTRACT OF FICUS RELIGIOSA IN ALLOXAN INDUCED DIABETIC RATS

Mamata L. Pochhi1, Dr. M. G. Muddeshwar2 1Department of Biochemistry, Swargiya Dadasaheb Kalmegh Smruti Dental College and Hospital, Nagpur 2Department of Biochemistry, Government Medical College, Nagpur

Objectives: The purpose of this study was to examine the hy- Methodology: The aqueous leaves extracts of Ficus religiosa poglycemic and antihyperglycemic effect of aqueous leaves (250 mg/kg) were administered orally for 21 days, in alloxan extract of F. religiosa in alloxan diabetes albino rats. induced diabetic rats. The effect of the leaves extracts of

123 S68 Ind J Clin Biochem

F.religiosa on Glucose, Lipid profile, Lipoproteins and Lipid activities of lipid metabolizing enzymes in hyperglycemic metabolizing enzymes were studied. rats. From this study it may be conclude that the extract acts by initiating the release of insulin by pancreatic beta Result: In alloxan induced diabetic rats, the aqueous cells of pancreas and having an extra pancreatic effect leaves extract of F. religiosa induced significant improve- to possibly increase the number of insulin receptors 1. ment in glucose, serum lipids, lipoproteins concentration ‘Ficus religiosa’, a traditional medicinal plant. This study and lipid metabolizing enzymes activity (Lipase, HMG CoA was performed to confirm the possible role of indigenous reductase and LCAT) with a single oral administration of medicinal plants in the treatment of Diabetes. F. religiosa. Keywords: Ficus religiosa, lipid profile, lipoproteins, LPL, Conclusion: The present study indicates that the aqueous HMG-CoA reductase, LCAT. leaves extract of Ficus religiosa exhibits lipid lowering, maintaining lipoproteins concentration and also improve the

IMPORTANCE OF ALBUMIN CREATININE RATIO IN SPOT URINE AS SCREENING TEST FOR EARLY DIABETIC NEPHROPATHY IN TYPE 2 DIABETES MELLITUS PATIENTS IN BIHAR

Dr. Rekha Kumari,1 Dr. Uday Kumar2, Dr. Anand Saran3, Dr. S.S. Haque4, Dr. Ratna Priya5, Dr. Shailesh Kumar6 1-5Department of Biochemistry, 6Department of Microbiology, Indira Gandhi Institute of Medical Sciences, Patna

Objective: The aim of this study is to detect diabetic Result: Albumin/Creatinine ratio in the control group was nephropathy at earliest stage in type 2 diabetic patients by 13.2 + 2.9 whereas in diabetic group it was 46.9 + 11.9 mg/ calculating albumin / creatinine ratio in spot urine sample, gm. Out of seventy diabetic cases 20 cases were having because rigourous control of blood sugar and medication albumin creatinine ratio less than 30 mg/gm but in rest at this stage can prevent progression of the disease to 50 case it was in the range of 30 – 300 mg/gm. Statisti- end stage renal disease. Its cut off range is 30 – 300 mg/ cal analysis was done. p value was found to be less than gm or 2.5-30 mg/mmol. 0.0001, which is extremely significant.

Method: We included 70 diagnosed cases of type 2 diabetes Conclusion: This study shows that albumin / creatinine mellitus of minimum 10 year duration attending medicine ratio can be used as screening test to detect early involve- department of IGIMS Patna from different places of Bihar. ment of kidney in type 2 diabetes mellitus cases in Bihar. 40 normal healthy persons of similar age and sex group Diagnosis of diabetic nephropathy at the earliest is very were taken as control. Any Other cause of proteinuria important because rigorous control of blood sugar and were excluded from both the groups. Estimation of urine addition of medication at this stage can prevent progres- albumin and creatinine was done separately and urinary sion of diabetic nephropathy. albumin/creatinine ratio were calculated in mg/gm unit on spot urine sample with commercial kit.

STUDY OF SERUM HSCRP AND PLASMA FIBRINOGEN LEVELS AS THE INDICATOR OF SEVERITY IN PREECLAMPSIA

Mrs Sonal Somali, Dr. (Mrs.) Poornima Dey Sarkar, *Dr. (Mrs.) Sumitra Yadav Department of Biochemistry, *Department of Obstetrics and Gynecology. M.G.M. Medical College, Indore

Objectives: To compare serum hs-CRP and plasma fibrino- healthy pregnant women (n=52) admitted in the obstetrics gen levels in mild preeclampsia women, severe preeclamp- and gynecology wards of M.Y. Hospital, Indore (M.P.). The sia women and normal pregnant women. blood samples were collected and analyzed for hs-CRP and fibrinogen. Study Design: The study was performed on three groups; mild preeclampsia (n=32), severe preeclampsia (n=18) and Results: In this study, it was found that the mean values

123 Ind J Clin Biochem S69

of serum hs-CRP in preeclamptic group were significantly group was 828.0 ± 53.0 mg/dl and in normal pregnant higher than the normal pregnant women. The mean value women was 557.0 ± 113.0 mg/dl. The plasma fibrinogen in mild preeclampsia group was 14.28 ± 11.62 mg/L, severe levels were significantly elevated in women with severe preeclampsia group was 34.0 ± 25.27 mg/L and in normal preeclampsia than mild preeclampsia. pregnant women was 3.42 ± 0.48 mg/L. The serum hs-CRP Preeclampsia is associated with a state of hypercoagulabil- levels were significantly elevated in women with severe ity and a systemic inflammatory response is considered to preeclampsia than mild preeclampsia. have a role in pathogenesis of preeclampsia and fibrinogen Normal pregnancy, a condition with an enhanced systemic being an important acute phase reactant, might therefore inflammatory response, becomes markedly exaggerated in correlates with preeclampsia and its severity. preeclampsia in response to excessive stimuli and triggers Conclusions: The present study showed that serum hs-CRP the endothelial dysfunction and inflammation leading to the and plasma fibrinogen levels have increased as disease maternal symptoms. The hs-CRP is a sensitive marker of progressed from mild to severe condition. Elevated lev- tissue damage and inflammation and plays an important els of serum hs-CRP and plasma fibrinogen reflects the role in eliciting the inflammatory processes. Its production involvement of inflammation in preeclampsia. Thus it was is stimulated by inflammatory cytokines, Interleukin-6 and concluded that hs-CRP and fibrinogen levels may be help- a-Tumor Necrosis Factor (α-TNF). ful to predict severity of disease. Similarly it was that the mean values of plasma fibrinogen Keywords: Preeclampsia, Norrnal pregnancy, Serum hs-CRP, in preeclamptic group were significantly higher than the Plasma Fibrinogen, inflammation. normal pregnant women. The mean value in mild preec- lampsia group was 724.0 ± 84.0 mg/dl, severe preeclampsia

ROLE OF RECENT CARDIAC BIO MARKERS IN THE DIAGNOSIS OF MYOCARDIAL INFARCTION AND THERE CUT OFF VALUE AT DIFFERENT INTERVALS IN GARHWAL POPULATION FROM A TERTIARY CARE CENTRE

Ajit Thakur#, Sukhes Mukherjee, Kailash Gairola, Sanjay bhatt, J.B. Gogoi Department of Biochemistry, VCSGGMC&RI and HNB Base Hospital, Srinagar, Pauri Garhwal, Uttarakhand

Mayocardial infarction ranks high in causing mortality the first two samples are negative and suspicion) and on worldwide and the Indian scenario is not much differ- 2nd day and was quantitatively analyzed by AQT 90 FLEX, ent. The recent markers like BNP, NTproBNP, D-dimer RADIOMETER, Denmark for following parameters – BNP, NT and hsCRP gives better edge from conventional markers pro BNP, D-dimer, hs CRP, Trop I, Trop T and CK-MB. like Trop I, Trop T, CK-MB, AST and LDH as they help in From the analysed results it was observed that there is diagnosis of Myocardial infarction, stroke and in various marked increase in the values of recent marker in support types of C.V.D. with conventional markers. The following study shows In our study, we have selected patients admitted in critical the significant elevation in values of D-dimer, BNP, NTpro condition in I.C.U. in H.N.B. BASE HOSPITAL, SRINAGAR BNP and hsCRP than the conventional markers like Trop with sign and symptoms of myocardial infarction (M.I.) I, Trop T, CK-MB, AST and LDH. The following pilot study and elevated S-T segment in ECG. 100 patients after dif- of recent cardiac biomarkers and after different time in- ferent time intervals were selected from last 5 months tervals gave us a different cut off range and peak values duration. 100 patients without any heart related problem after M.I. and helps the clinicians in monitoring C.V.D as coming for comprehensive check up in medicine depart- even electrocardiography is not able to detect the cardiac ment was selected as control. The blood sampling is serial profile changes at this span of time. This is the first study in order to detect a rise and /or fall of the biomarker i.e. of such type in the Garhwal population where we have at admission, 6 to 9 hours (optionally 12 to 14 hours if highlighted some significant aspects of the latest cardiac

123 S70 Ind J Clin Biochem

biomarkers and there cut off values in one of the most Keywords: M.I., BNP, NTpro BNP, D-Dimer, hsCRP, CK- MB, prominent disease causing maximum mortality and morbid- AST, Trop I and Trop T ity amongst various age groups.

SERUM URIC ACID LEVEL & OXIDATIVE STRESS OF PSORIASIS PATIENTS IN RELATION TO DIABETES MELLITUS AND HYPERTENSION

Dr. Payasvi Baweja1, Dr. B. K. Agarwal2, Dr. V.K. Sharma3, Dr. Ana Alex4 1Department of Biochemistry, GMC Bhopal, 2Ex. Prof. & HOD Department of Biochemistry, GMC Bhopal, 3Prof. & HOD Department. of Medicine, GMC Bhopal, 4Prof. & HOD Department. of Skin, GMC Bhopal

Object: To measure newer diagnostic tests in Psoriasis groups, Group II with only psoriasis and Group III with patients in relation to D.M./H.T. & to prevent progression psoriasis patients suffering from DM/HT and 50 age and of co-morbidities. sex matched healthy control subjects in Group I. In all groups estimation of serum SOD done by manual method Aim: For determination of level of lipid peroxidation as (Mishra HP and Fridovich I), serum TAC (D. Koracevic et (Malondialdehyde MDA) and antioxidants (Total antioxi- al method 2001), plasma MDA (by thiobarbituric acid as- dant status, superoxide dismutase SOD) and its co-relation say Buege et al 1978). Serum uric acid by colorimetric with the severity of psoriasis and to determine serum method at 546nm. uric acid level. Results: The mean ± S.D. of MDA, SOD, TAC, Serum uric Material & Method: The present study comprised of 50 acid of all groups and their significance is summarized in clinically diagnosed psoriasis patients divided into two table shown below:

Parameters Units Control Group I Psoriasis Group Psoriasis with * P<0.001 as compared to Group I II DM/HT Group III # P<0.001 as compared to Group II No. Of Subjects 50 25 25 @ P>0.05 as compared to Group II P. MDA n mol/ml 0.91±0.22 1.73±0.27* 3.98±0.17# i.e. P. MDA is significantly elevated in both groups of S. SOD U/gm Hb 6.80±0.76 5.39±0.32* 3.28±0.24# patients as compared to controls. S. TAC μ mol/lit 1752.21±16.69 1621.27±16.88* 1067.83±23.37# S.SOD & S.TAC is significantly decreased in both groups of patients as compared to controls. S. Uric Acid mg/dl 4.80±0.16 7.20±0.64* 7.80±0.64@

INFLUENCE OF OXIDATIVE STRESS ON PROTEIN CARBONYL CONTENT AND GLYCEMIC STATUS IN TYPE II DIABETES

Pradeep Kumar Vegi, Dayanand C. D, A. V. M Kutty, Krishna Murthy N Dept. of Biochemistry& Allied Health Sciences, Sri Devaraj Urs Academy of Higher Education and Research, Kolar

Aim: On the basis of increased oxidative stress observed estimated according to method described by Levine et al. in diabetes mellitus, the present work is designed to de- using 2, 4 di nitro phenyl hydrazine (DNPH), which reacts termine its effect on plasma protein measured by Protein with protein carbonyls forming a Schiff base to produce the carbonyl content and glycemic status which serves as hydrazone, measured spectrophotometrically at 370nm. biomarker of oxidative protein damage in type II diabetes Results: The Mean±SD levels of total protein carbonyl in comparison to healthy controls. content nmol/ml (0.70±0.34), fasting plasma glucose mg/ Materials and Methods: 60 Diabetic subjects, 60 healthy dl (79.12±11.74), HbA1c % (5.79±0.66) and plasma insulin controls attending to R. L. Jalapa hospital, Kolar were mcu/ml (13.16±11.10) in control group and (1.68±0.47), recruited in the study. Plasma Protein carbonyls were (182.58±102.42), (8.86±2.24), (18.61±17.15) respectively in

123 Ind J Clin Biochem S71

diabetic group. There is a significant positive increase re- Conclusion: The above findings suggeststhe a deleterious flected between control and diabetic group with respect effect of oxidative stress on total plasma protein which to protein carbonyl groups. Similar observation noticed is reflected by increased level of protein carbonyl content with respect to glycemic status in terms of HbA1c and along with glycemic status which can lead to complications plasma glucose, with a normal insulin levels in both the observed in Type II diabetes mellitus. study group. Keywords: Protein carbonyl, Insulin, DNPH, protein marker, Oxidative stress.

EVALUATION OF OXIDATIVE STRESS IN OBESE MEDICAL STUDENTS WITH REFERENCE TO CARDIOMETABOLIC SYNDROME

Meghana K Padwal, Priya Kamthe, R.R. Melinkeri Associate Professor, Post Graduate Student, Professor and Head, Department of Biochemistry, BVDUMC Pune

Background: The prevalence of obesity in young population levels were done by automated biochemistry analyser. is on rise. The major complication of obesity is dylipedemia, Estimation of hs-CRP was done by automated chemilumi- impaired glucose metabolism etc. It has been demon- nescence immunoassay system. strated that fat accumulation correlates with markers of Results: There was statistically significant rise in the levels systemic oxidative stress. Oxidative stress may have pro- of hs-CRP and MDA in obese students as compared to non inflammatory role on promoting atherosclerosis. obese (p=0.001). There was no statistically significant dif- Objectives: The present study was designed to evaluate ference in the levels of blood glucose in both the groups the relationship between the marker oxidative stress i.e. (p = 0.06). However the levels of Total cholesterol, LDL Malondialdehyde (MDA) with reference to markers of were significantly increased in cases (p = 0.01), while Cardiometabolic Syndrome i.e. hs- CRP, lipid profile and levels of HDL were significantly decreased (p = 0.01) as blood glucose in healthy obese students. compare to controls.

Material and Methods: The present study was a prospec- Conclusions: There is increased oxidative stress in obesity tive case control study, carried out in a university medical which may aggrevate the proinflammatory processes of college. The participants were healthy medical students of atherosclerosis. The metabolic changes in obesity are asso- both the genders. They were divided into cases: Obese ciated with to chronic systemic low grade inflammation. (BMI ≥ 25 kg/m2; n=30) and controls: Non obese (BMI ≤ Keywords: Oxidative stress, Cardiometabolic Syndrome, 25 kg/m2; n=30). Estimation of MDA, Lipid Profile, glucose MDA, hs – CRP.

ASSESSMENT OF UMBILICAL CORD TSH LEVELS IN SMALL FOR GESTATIONAL AGE NEONATES

*M. K Padwal, *B.D. Kamble, *R.R. Melinkeri, P.B. Suryawanshi** Associate Professor, JRII, Professor & Head, Associate Professor, *Department of Biochemistry, **Department of Paediatrics, BVDU Medical College, Pune

Background: Small for gestational age (SGA) neonates Objectives: The present study was a prospective cross- are born as a result of Intrauterine Growth Retardation sectional cohort study, designed to assess Umbilical cord (IUGR). This may result due to complex factors of fetal, Thyroid Stimulating Hormone (TSH) levels in a term small placental and maternal origin. Thyroid hormones are for gestational age neonates. critical for growth and development of the fetus. It may Material and Methods: A group of term babies delivered permanently influence the endocrine system by affecting to eu-thyroid mothers was selected from a tertiary care its programming during development.

123 S72 Ind J Clin Biochem

teaching hospital. It was subdivided into cases (SGA Conclusions: The finding of higher TSH concentration in a neonates with birth weight ≤ 2.5 kg, n=58) and control term SGA neonates suggests that there may be intrauter- (Average for gestational age neonates (AGA) with birth ine reprogramming of TSH sensitivity at the hypothalamic weight ≥ 2.5 kg, n=66). Umbilical cord TSH levels were pituitary level. However a larger survey with an appropriate measured in SGA and AGA neonates by chemiluminescence follow-up is required to evaluate the prevalence and degree immunoassay system. of thyroid dysfunction in children born in a term SGA.

Results: There was statistically significant rise in cord TSH Keywords: Small for Gestational Age, IUGR, TSH. levels in SGA neonates as compared to AGA (P=0.001).

ANTIHYPERLIPIDEMIC AND ANTIOXIDANT EFFECT OF AQUEOUS EXTRACT OF WITHANIA COAGULANS FRUIT IN CHOLESTEROLFED HYERCHOLESTEROLEMIC RABBITS

Kirtikar Shukla, Piyush Dikshit, Rimi Shukla, Jasvinder K Gambhir Department of Biochemistry, University College of Medical Sciences (University of Delhi) and GTB Hospital, Delhi

Antihyperlipidemic and antioxidant effects of Withania (GSH), superoxide dismutase (SOD) and ferric reducing coagulans (aqWC) have been studied in cholesterol-fed ability of plasma (FRAP) (p<0.05 vs Gp I). Cholesterol + rabbits. Animals were divided into three groups. Group I; aqWC administered (group III) animals showed significantly healthy controls, group II; cholesterol-fed (100 mg/kg bw) decreased levels of TC, TAG, LDL-C and tissue lipid content and group III; Cholesterol + aqWC (250 mg/kg bw) for six and increased HDL-C levels (p<0.05 vs Gp II). HMGCR and weeks. Cholesterol feeding led to significant increase in LPL activity increased significantly whereas ACC activity total cholesterol (TC), triacylglycerol (TAG), low density decreased in group III animals (p<0.05 vs Gp II). Group lipoprotein (LDL-C) and tissue lipid content and decrease III also showed significantly decreased MDA and increased in high density lipoprotein (HDL-C) (p<0.05 vs Gp I). Activ- GSH, SOD and FRAP activity (p<0.05 vs Gp II). These results ity of HMG CoA reductase (HMGCR) and lipoprotein lipase suggest that the aqWC may have beneficial effects on lipid (LPL) decreased whereas acetyl CoA carboxylase (ACC) profile and activity of enzymes of lipid metabolism. It also activity increased in group II (p<0.05 vs Gp I). Group II ameliorates oxidative stress by improving antioxidant sta- animals also showed significantly increased lipid peroxide tus. These effects may improve lipid profile and decrease (MDA) and decreased antioxidants i.e. reduced glutathione oxidative stress in hyperlipidemic state.

STUDY OF INSULIN RESISTANCE AND LIPID PROFILE IN OBESE ADULTS

Arti Gurung*, Prof. Dr. Ebenezer William, Dr. Meera shivsekhar Department of Biochemistry, SRM medical College Hospital and research centre, SRM University, Kattankulathur

Introduction: Insulin resistance and abnormal lipid profile insulin resistance (IR) index was calculated and the ratio of are the risk factors for cardiovascular disease in obesity. lipid was calculated. All statistical analysis were performed The aim of the study is to determine the level of insulin using the statistical package SPSS. resistance and lipid profile in obese Indian adults. Result: According to this study, the levels of fasting se- Method: About 35 obese individuals between the age rum insulin, Total cholesterol (TC), Low density lipoprotein group 18-50 yrs from the out-patients attending SRMMCH (LDL-C), were higher in obese than in controls while high & RC for general check up and an equal number of healthy density lipoprotein cholesterol (HDL-C) and insulin resis- controls were selected. Anthropometric and plasma bio- tance were lower in the obese. chemical variables (including fasting glucose, HbA1c, insulin Conclusion: The study reveals that there is a strong re- and lipid profile) were measured by standard methods. lationship between insulin resistance and lipid profile in By using homeostasis model assessment (HOMA) method obese adults.

123 Ind J Clin Biochem S73

DISCRETE BANDS SEEN AT THE POINT OF APPLICATION IN ALL LANES OF IMMUNOFIXATION ELECTROPHORESIS

Rajeev Gupta, Dilip Kumar, Poonam Das, Nitin Dayal, Anupama Arya, Andleeb Abrari, Twinkle Garg Institute of Laboratory Medicine, Max Super Specialty Hospital (West Block), New Delhi

Objective: To analyze a rare case of discrete bands seen Results: Biclonal ‘M’ spike was seen in Gamma region of at the point of application in all lanes of Immunofixation SPE lane. In all other lanes (i.e. IgG, IgM, IgA, K, λ) a dis- Electrophoresis. crete Band was seen at the Point of application. Biclonal ‘M’ Band seen in Gamma region was of IgM kappa type. Material and Method: A serum sample from 61 years male Total protein was 10.4 g/dL. IgM was very high 5525 mg/ patient was requested for serum Protein Electrophoresis dl {60-280}. Serum Beta 2 Micro globulin was 8.44 mg/L (SPE) and Serum Immunofixation Electrophoresis (IFE). {0.9-3.0}. Rheumatoid Factor (RF) was negative. Bence The SPE and Serum IFE were performed on agarose Jones Protein was negative. Histo Biopsy and Immuno- gel kit provided by Helena Biosciences. The Electropho- histochemistry (IHC) from Left Axillary lymph node was retogram was scanned by scanner and the densities of reported as Lympho-plasmacytic Lymphoma (Low grade the discrete bands were measured at 620 nm by using B- cell Non Hodgkin Lymphoma). Platinum software, Helena Biosciences. Serum Protein was analyzed by DXC 800 Auto analyzer, Beckman Coulter. In Conclusion: Final Diagnosis of Waldenstrom’s macroglobu- addition Immunoglobulin Profile (IgG, IgA and IgM), Beta linemia was made. The discrete Band seen in all lanes of 2 Micro globulin, 24 hours Urine IFE, Bence Jones Protein, Serum IFE was due to non specific staining of precipitated Complete Blood Count (CBC), Bone marrow Aspirate and IgM proteins which got adhered to gel matrix of Agarose Biopsy, Flow Cytometry for CLL and Left Axillary lymph and was stained by Acid Blue Stain. node biopsy with IHC were also done.

EVALUATION OF NITRIC OXIDE AND TRACE ELEMENTS IN BREAST AND OTHER CANCER PATIENTS

Anjali Vagga*, Anita Kale, N.R. Akarte and Kumud Harley *Assistant Professor, Department of Biochemistry, JNMC, SAWANGI (M), Wardha

Aim: To determine the alteration in total nitric oxide level was more in other cancer patients as compared to (NO), nitrate (NO3-), nitrite (NO2-), Copper (Cu) and zinc the breast cancer patients (p<0.05). Serum NO3-levels (Zn) in serum and Cu/ Zn ratio and role they may play in were significantly more in other cancer patients (p<0.0001) contributing to breast and other cancers. while NO2-level were raised in breast cancer patients (p<0.0001) as compared to the healthy controls. There was Material and methods: The present study was conducted a significant fall in serum NO2- in other cancer patients in total of 90 subjects divided into three groups as as compared to the healthy controls and breast cancer control (n=30), breast cancer (n=30) and other cancers patients (p<0.05, p<0.0001 respectively). (n=30). Serum NO and NO3 was estimated by cadmium reduction method. Serum Cu and Zn were estimated by Conclusion: Serum levels of NO, NO3-, NO2-, Cu and Zn atomic absorption spectrometry. Serum NO2- and Cu/ Zn and recently application of the Cu/ Zn ratio can be used ratio were calculated. as potentially useful parameters in evaluating extent of disease, disease activity and prognosis in patients with Results: Total NO, Cu and Cu/Zn ratio were significantly variety of cancers. raised (p<0.0001) while Zn levels were significantly low (p<0.0001), in serum of both breast and other cancer Keywords: Atomic absorption spectrometry, nitric oxide, patients as compared to the healthy controls. Fall in Zn nitrate, nitrite, cadmium reduction.

123 S74 Ind J Clin Biochem

EVALUATION OF ANTIOXIDANT ENZYMES & LIPID PEROXIDATION IN PATIENTS WITH TYPE 2 DIABETES MELLITUS WITH OR WITHOUT NEPHROPATHY

Manjulata Kumawat1, Ishwar Singh3, Tarun Kumar Sharma1, Neelima Singh2, Veena Singh Ghalaut1, Kiran Dahiya1, Ramanjit Kaur1 1Department of Biochemistry, Pt. B.D. Sharma University of Health Sciences, Rohtak Haryana, 2Department of Biochemistry G.R. Medical College, Gwalior (MP), 3Department of Neurosurgery, Pt. B.D. Sharma University of Health Sciences, Rohtak Haryana

Background: Oxidative stress has been considered to be Creatinine, Triglyceride (Tg), VLDL-Cholesterol (VLDL-C), a pathogenic factor of diabetic complications including Catalase (CAT) and Malondialdehyde (MDA) were signifi- nephropathy. Hyperglycemia and dyslipidemia in diabetes cantly elevated in Type 2 diabetic patients with and without mellitus induce increased lipid peroxidation and peroxyl nephropathy as compared to controls and also significantly radical formation is an important mechanism in genesis of higher in Type 2 diabetic patients with diabetic neph- micro-angiopathy. There are many controversies regarding ropathy as compared to Type 2 diabetic patients without the status of antioxidant enzymes in diabetic nephropa- nephropathy. Activities of reduced glutathione, Superoxide thy also limited studies are seen which include all these dismutase, glutathione peroxidase, Glutathione reductase, antioxidant enzymes, so we planned this study by taking were significantly lower in type 2 diabetic patients with most of the antioxidant enzymes. and without nephropathy as compared to controls and also significantly lower in Type 2 diabetic patients with diabetic Methods: The study group included 90 subjects, out of nephropathy as compared to Type 2 diabetic patients which, 60 patients with Type 2 Diabetes Mellitus (30 each without nephropathy. There was significant correlation with and without nephropathy) and 30 healthy controls between all these parameters with GSH and MDA within age group of 45-65 years. Blood samples of all subjects were collected and analyzed for glycosylated He- Conclusion: These results confirm that intensity of oxida- moglobin, Fasting and post prandial plasma glucose, Urea, tive stress in type 2 diabetic patients with nephropathy Creatinine, Total cholesterol, Triglyceride, HDL-Cholesterol, is greater when compared with type 2 diabetic patients LDL-Cholesterol, VLDL-Cholesterol, reduced glutathione, without nephropathy as compared to the control group. glutathione peroxidase, Glutathione reductase, catalase, Keywords: Non insulin dependent diabetes mellitus, Anti- Superoxide dismutase and Malondialdehyde in Type 2 oxidant enzymes, Malondialdehyde, reduced glutathione, Diabetes Mellitus patients with or without Nephropathy. lipid profile, glycosylated Hemoglobin and diabetic neph- Results : Glycosylated Hemoglobin (HbA1c), Fasting plasma ropathy. glucose (FPG), post prandial plasma glucose (PPPG), Urea,

STUDY OF OXIDATIVE STRESS AND NEW EMERGING CARDIAC RISK MARKERS IN SUBCLINICAL AND OVERT HYPOTHYROIDISM: BEFORE AND AFTER TREATMENT

Sarika Singh Chauhan and Dr. Purnima Dey Sarkar Department of Biochemistry, NSCB (Jabalpur) & MGM (Indore) Medical College

Objective: Overt (OH) and subclinical (SCH) hypothyroid- Methods: 50 SCH, 50 OH and 100 euthyroid control ism are associated with dyslipidemia, an increased level patients of similar age and sex, were enrolled for the of low-grade inflammation, and oxidative or antioxidative study. In these patients’ serum concentrations of fT4, status of the organism. Hyperhomocysteinemia is an im- fT3, TSH, lipid parameters (total cholesterol, triglyceride, portant, independent risk factor for atherosclerosis and HDL-C, LDL-C), C-reactive protein, homocysteine, MDA and thrombotic disease. This study was designed to evaluate Paraoxonase levels were assessed, before and after 8-12 oxidative stress and new emerging cardiac risk markers weeks of treatment. in SCH and OH before and after treatment. • Fasting Lipid profile and CRP by fully auto analyzer.

123 Ind J Clin Biochem S75

• Homocysteine and Serum level fT3, fT4 & TSH by (P<0.01) and lower fT3, fT4, HDL levels. L-thyroxine therapy chemiluminescence assay. significantly decreased serum TSH (P<0.001), TC, LDLc, TG, • MDA and Paraoxonase by spectrophotometer. CRP (p<0.01) and homocysteine (P<0.001) concentrations in both groups. Results: Pretreatment, in both SCH and OH groups, there were significantly higher serum TSH (P<0.001), TC (P<0.05), Conclusion: These results suggest that patients with SCH LDL-C (P<0.01), hs-CRP (0.01) and MDA (0.05) levels, than and OH exhibited elevated atherogenic parameters and in controls. The activity of PON-1 was significantly lower high risk of cardiovascular diseases. Lipid peroxidation may than controls (P <0.05). No significant changes were found have a role in the pathogenesis of the atherosclerosis in in other parameters in SCH patients compared to controls both hypothyroidism. Thyroid hormone replacement ther- and overt hypothyroid patients whereas OH patients apy may slow the progression of coronary heart disease, also showed significantly higher serum TG, homocysteine because of its beneficial effects on lipids.

HYDROXYL RADICAL GENERATION BY CATALASE: A FEW EXPERIMENTAL EVIDENCES

Madhur M. Goyal, Ajay Meshram & Anjan Basak Department of Biochemistry, J.N. Medical College, DMIMS (DU), Wardha

Catalase is well known antioxidant enzyme which protects ence of azide or urea induced structural changes of BLC cells from the toxic effects of hydrogen peroxide. Unlike suggest that the enzyme participate in the production of peroxidase, it catalyses the dissociation of hydrogen perox- HRs. The possible character is unusual in contrast to the ide directly into H2O and O2. Our results show that apart well established fact that catalase reduce the generation of from the known catalytic action, catalase can generate reactive oxygen species in biological system. The possible Hydroxyl Radicals (HRs) in experimental conditions. At low new character (generation of HRs) could be an answer of concentrations of H2O2 (<20μM), presence of bovine liver several unexplained features of catalase, however, need catalase (BLC) increased the generation of HRs in reaction to be tested further. mixture. Differential productions of HRs due to either pres-

COMPARE SERUM CONCENTRATIONS OF NUTRIENT ANTIOXIDANT VITAMINS, VIZ., VITAMIN C AND E IN SUBJECTS DIVIDED ACCORDING TO BMI AND TO FIND ASSOCIATION OF VITAMIN C AND E LEVELS WITH METS IF ANY.

Neelam Khandelwal Govt. Medical College, Kota

Object: Experimental and clinical observations indicate three groups - Normal, Overweight and Obese consisting oxidative stress and depletion of antioxidants as an im- of 40 subjects in each group. Data were analysed using portant pathogenic mechanism in obesity-associated meta- students‘t’ test, two ways ANOVA and strength of associa- bolic abnormalities and in the development of DM and tion between two variables was measured by Pearson’s its complications. This study is aimed to compare serum correlation coefficient (r). concentrations of nutrient antioxidant vitamins, viz., vitamin Results: Serum vitamin C and E concentrations were C and E in subjects divided according to BMI andto find significantly lowered in obese subjects as compared to association of vitamin C and E levels withMetS if any. overweight and normal subjects (f = 51.27, P < 0.0001 & Method: Study includes 120 male subjects ranging in age f = 32.25, P < 0.0001 respectively for vitamin C and E). 30-50 years. All the subjects were screened for obesity Subjects with MetS have significantly lower concentrations according to new guidelines given by the Indian Ministry of vitamin C and E as compared to those without MetS of Health. Efforts were made to divide the subjects into in all the three groups (p < 0.01).

123 S76 Ind J Clin Biochem

Conclusion: Vitamin C and E levels were found to have association between increased abdominal fat storage, and anegative association with BMI (r = -0.6687 and r = -0.5834 systemicoxidative alterations in MetS patients. respectively) and with MetS. In conclusion there is a strong

STUDY OF THE ASSOCIATION OF LIPID LEVELS AND PROTHROMBOTIC STATE IN DIABETES AT A TERTIARY CARE HOSPITAL IN CENTRAL DELHI

Bhawesh Mishra, Ekta Malik, Anju Jain Deptt. of Biochemistry, Lady Hardinge Medical College, New Delhi

Background: Diabetics are known to be at increased risk were reported as Mean ± SEM. Independent t-test was of developing cardiovascular diseases probably due to un- applied to compare means. Pearsons correlation analysis derlying prothrombotic state caused by hyperglycemia and was performed to determine the association between dyslipidemia. An increased Plasminogen Activator Inhibitor- different variables. 1(PAI-1) level predisposes to a prothrombotic state and is Results: Serum PAI-1 levels were significantly higher in responsible for adverse cardiovascular outcomes. PAI-1 is diabetics (165±10.9 vs. 114±8.1ng/ml, p<0.001). Serum the primary inhibitor of plasminogen activator which plays triglyceride, cholesterol, VLDL and LDL showed a posi- a major role in fibrinolysis. tive correlation with PAI-1 levels whereas HDL showed Objectives: The study aimed at estimating blood lipids negative correlation. Blood glucose (r=0.7), triglyceride and PAI-1 levels in diabetic and non-diabetic subjects and (r=0.53) and VLDL (r=0.53) correlated more strongly with further correlate levels of blood glucose and lipids with PAI-1 level compared to total cholesterol (r=0.36), HDL PAI-1 levels to study the association. (r=-0.29), LDL (r=0.27).

Methods: 30 known diabetic cases and 30 age & sex Conclusion: Serum PAI-1 levels are raised in diabetics, matched apparently healthy non-diabetic controls were predisposing to a prothrombotic state. Blood glucose and randomly enrolled for the study. Subjects suffering from triglycerides strongly correlated with PAI-1 levels. Hence an other chronic diseases, inflammatory conditions or im- early intervention by good glycemic control and reducing paired liver and/or renal function were excluded. Fasting triglyceride levels can prevent cardiovascular complications serum lipids, blood glucose and Plasminogen Activator in diabetics. Inhibitor-1 levels were estimated in all subjects. The data

EVALUATION OF BIOCHEMICAL INDICES IN CHRONIC ALCOHOLISM

Akanksha Dubey, Dr B.K. Agrawal, Dr Bhawna Bhimte Department of Medical Biochemistry, GMC Bhopal

Objectives: Chronic alcoholism perturb to pathological drinker’s (90.37±9.09 & 127.61±15.33 respectively) and dura- condition affecting the nervous, gastro enteric system. tion 8-10years with mean of 9.4±0.74 and >10years with Hepatocytes are particularly susceptible to alcohol me- average of 12.26±1.46 of alcohol. Biochemical parameters tabolism; consequently abnormalities could be recurrent in estimated were liver enzymes (GGT, AST, ALT and AST/ liver function test. Present study has been undertaken to ALT). AST-ALT were done by U-V Kinetic method and GGT evaluate various biochemical alterations caused by alcoholic by End Point Reaction method. beverages on long term ingestion. Result: Mean ±SD values of GGT for alcoholics were Methodology: Study has been carried out on 50 alcoholic 167.5±95.67 (p <0.001) as compared to controls morbid groups of 20-60 years of age compared to 50 age (14.01±0.613). The ratio of AST/ALT showed significant matched non alcoholic control. Morbid group are again raise in alcoholics than the controls (1.35±0.186, 1.02±0.005 classified on the basis of amount Moderate and heavy respectively). Parallel results were also observed with

123 Ind J Clin Biochem S77

Moderate drinkers compared to heavy drinkers. marker of alcohol abuse. However detailed studies with large sample size may be required to establish LFT as Conclusion: Evaluation of hepatic parameters establish that marker of alcoholism. alcohol interfere with liver functioning. The data suggest that elevations of enzymes can be a particular and specific

LIPID PEROXIDATION IN CORD BLOOD AT BIRTH A COMPARATIVE STUDY OF NORMAL VAGINAL DELIVERY AND CAESARIAN SECTION

Neelima Singh, Amrita Richhariya, Ekankshi Sharma, Sanjeev K. Singh, Nivedita Singh, Vishal Bhargava Department of Biochemistry, G.R. Medical College and J. A. Group of Hospitals, Gwalior

Background and objective: Pregnancy is a stressful condi- eters were estimated, MDA as marker of lipid peroxidation tion often accompanied by a high energy demand of many and SOD as antioxidant enzyme. bodily functions. Genetic variability, maternal oxidative Result: Highly significant (p<0.001) increased levels of MDA stress, maternal antioxidant capacity are likely to alter & decreased levels of SOD in the cord blood of foetuses the cord blood lipid peroxidation and antioxidant enzyme. born via normal vaginal deliveries as compared to cesarean During labour birth, increased oxidative stress will results section indicates a condition of oxidative stress in former in the increased oxidation of macromolecules in the tissue group i.e. Normal vaginal delivery subjects. damage. This study was aimed to estimate the level of lipid peroxidation and antioxidant enzyme in the umbilical Conclusion: We concluded that oxidative stress is more in cord blood of newborn infants delivered through caesarean the cord blood of foetus born via normal vaginal delivery section or normal vaginal delivery. therefore it is more stressful for both mother & the new- born which signifies the importance & safety of elective Materials and methods: Study has been carried out in caesarean section in certain cases. 50 female subjects, divided into two groups, 25 female subjects with normal vaginal delivery & 25 female subjects Keywords: Lipid peroxidation, antioxidant, cord blood, with caesarean section. In both groups following param- Oxidative stress.

KIDNEY DYSFUNCTION IN SICKLE CELL DISEASE POPULATION

Gopinath Agnihotram*, Dr. Purnima Dey Sarkar**, Lincy K. Skaria* *Research Scholar in Dept. of Biochemistry, M.G.M. Medical College, Indore, **H.O.D., Dept. of Biochemistry, M.G.M. Medical College, Indore

Objective: To study and compare several types of Kidney lege, Indore. Complete blood count, urinalysis, along with dysfunctions in Sickle Cell Disease (HbSS) and Sickle Cell Serum Creatinine estimation by the Jaffe’s reaction, Total Trait (HbAs) population. Protein estimation by Biuret method, Creatinine clearance determinations were based on 24-h urine collection and Materials and Methods: A cross sectional randomized were calculated as (Urine Creatinine × 24-h Urine vol- study conducted on 180 individuals (n=100) of HbSS ume) / Plasma Creatinine, Serum Bilirubin estimation by and remaining (n=80) belongs to HbAS. The patients Zendrassik Method, Serum Lactate Dehydrogenase (LDH) included both male and female around the age group by Enzymatic kinetic method, Serum Electrolytes by Iso of 5-56 years. The detailed clinical examination has done Selective Electrode automation. on each individual with recorded demographics, medical and family history. The collected samples are processed Results: Serum Creatinine (<0.01) is not significant in at clinical Biochemistry Laboratory of M.G.M. Medical Col- comparison of both groups, Urinary Osmolality (<0.001) is

123 S78 Ind J Clin Biochem

significantly reduced in HbSS population with comparison of renal pathology has been disrupted and causes glomerular, HbAS, Micro Albuminuria (<0.001) is significantly increased tubular changes such as mainly Protenuria, Hyposthenuria, in HbSS, Serum Bilirubin (<0.001) and Serum LDH (<0.001) Hematuria, Nephritic syndrome, Renal failure. are significantly increased in HbSS with comparison to Conclusion: We thus can conclude that the polymeriza- HbAS, the remaining parameters are more or less similar tion of HbS is not confined to any one region of kidney or equivalent in both the groups. it affects all the parts including vasa recta. Increased risk Discussion: The kidney is sensitive to the vasoocclusion with increment in age and Hyposthenuria is the prominent induced hypoxia that can result from red cell sickling and consistent feature bserved in kidney abnormalities and from sickle cell endothelial cell adhesion. Acidosis, in relation to sickle cell disease. Raised hemolytic factor, Hypertonicity and Hypoxia are the factors tend to pro- Proteinuria, Hypertension are the parameters predict End mote HbS polymerization and red cell sickling there by stage renal disease. changes in oxygen delivery to kidney, By this reason the

THE VARIATION IN SECRETION OF GONADOTROPIN HORMONES AND CARDIOVASCULAR RISK IN PATIENTS OF POLYCYSTIC OVARIAN SYNDROME

Jaya Thilakan*, Dr B.K. Agrawal*, Dr Shabana Sultan**, Dr Bharti Parihaar**, Dr Tripti Saxena*, Dr Bhawana Bhimte* Department of Medical Biochemistry* Department of Obsterics and Gynecology** GMC Bhopal

Introduction: PCOS is most common endocrine disorder FSH was estimated by CLIA method, total cholesterol by affecting female reproductive system and is one of the CHOD-PAP method, triglyceride by GPO-PAP method leading causes of infertility. It is characterized by a complex Result: Total cholesterol, TG and LDL were significantly physiology implicating an interaction with environmental higher in PCOS patients while HDL levels were very low. and genetic factors, resulting in broad spectrum of repro- Also in about 70% patients LH/FSH ratio were greater ductive and metabolic disorders. than 2. Aim: Study of LH/FSH ratio and lipid profile in PCOS Conclutions: Increased LDL, TG and lower levels of HDL patients shows that women suffering from PCOS are at higher risk Methodology: Study has been carried out on 50 PCOS of cardiovascular disorders; also increased LH/FSH ratio patients and 50 controls of age group 16-30 years. LH and signifies inappropriate gonadotropin secretion.

ANALYSIS OF CARDIOVASCULAR RISK IN HYPERTENSIVE AND MYOCARDIAL INFARCTION

Saman Amin, Dr. Arun Shrivastav, Dr. Tripti Saxena, Dr. B.S. Yadav, Dr. Bhawna Bhimte Department of Medical Biochemistry, Gandhi Medical College, Bhopal

Introduction: Hypertension or high blood pressure is a prolonged ischemia. This is usually result from an imbal- chronic medical condition in which the blood pressure ance in oxygen demand, resulting in an acute reduction in the arteries is elevated this require the heart to work with thrombus formation in a coronary vessel, tresulting harder than normal to circulate blood through the blood in an acute reduction of blood supply to a portion of vessels it involves 2 measurements systolic and diastolic. myocardium. Hypertension and myocardial infarction are normal blood pressure is at or below 120/80 mm hg Hy- closely linked in several ways pertension is a risk factor to stroke myocardial infarction Method: The aim of this study was the assessment of lipid Myocardial infarction commonly known as heart attack it profile and c-reactive protein level in hypertension and is the irreversible necrosis of heart muscle secondary to

123 Ind J Clin Biochem S79

myocardial infarction patients the present study includes Result: The study showed the significant increased in lipid 100 patients of hypertension and myocardial infarction profile and significant increase in crp levels. patients and 100 healthy control blood sample where tak- Conclusion: In conclusion, increased lipid profile, and ing and following investigation were done- estimation of increased crp levels is associated with hypertensive and lipid profile by GPO/POD, CHOD-PAP method. Estimation myocardial infarction patients. of c-reactive protein by LATEX SLIDE METHOD.

EVALUATION OF THE FERTILITY POTENTIAL IN MEN WITH SICKLE CELL DISEASE

Prashant Nigam*, Dr. Purnima Dey Sarkar**, Vikas Gupta*, Dr. S.K.B. Patil* Dept. of Biochemistry, CIMS, Bilaspur (C.G.), **Dept of Biochemistry, MGMMC, Indore

Infertility is a major problem in Sickle Cell Disease Pa- cases 20±9, p<0.05), lower mean serum total testosterone tient, especially in males. In order to assess the fertil- (Control 7.2 ± 1.8, Cases 4.4±2, p< 0.01) in SCD subjects ity potential in men with Sickle Cell Disease serum LH, than in normal control. The fertility potential associated FSH, Prolactin & Total Testosterone were determined by with lower testosterone in subjects with SCD suggested using Radio Immunological technique. In present study that the hypothalamic/pituitary function in these patients 20 males with SCD & 20 normal control subjects were is intact and that the primary fault leading to poor sperm used. Our results shows a significantly higher mean FSH production by the testes. (Control 12±4, Cases 16.2±3), & Prolactin (Control 11.4±4.6,

INDUCED BIOCHEMICAL CHANGES IN ASTHMATIC PATIENTS

Swati Pathak*, Dr. B.K. Agarwal*, Dr. Lokendra Dave**, Dr. Tripti Saxena*, Dr. Bhawna Bhimte* Department of Biochemistry*, Department of Chest Medicine**, Gandhi Medical College, Bhopal

Introduction: Asthma is a disorder caused by inflammation CRP estimated by Latex slide test, P-MDA estimated by in the airways (called bronchi) that lead to the lungs. This Jean C.D. et.al. and Serum-SOD estimated by Mishra H.P.& inflammation causes airways to tighten and narrow, which Fridovich I method. blocks air from flowing freely into the lungs, making it Result: Increased CRP value (p<0.001) was observed in hard to breathe. Symptoms include wheezing, breathless- asthmatic patients. Also a significant increase in P-MDA ness, chest tightness and cough, particularly at night or (p<0.001), a decrease in S-SOD (p<0.001) & serum magne- after exercise/activity. sium level (p<0.001) were observed in asthmatic patients Objective: The study was designed to document changes as compared to normal healthy subjects. in serum magnesium level, serum CRP level and lipid per- Conclusion: Oxidative stress has a role in the pathogenesis oxidation and antioxidant status in asthmatic patients. of asthma. Oxidative stress aggravates airway inflammation Methodology: 50 asthmatic patients of age group 20-50 by inducing diverse proinflammatory mediators, enhancing years were enrolled for the study and were compared to bronchial hyperresponsiveness, stimulating bronchospasm, 50 normal healthy individuals of same age. Those fulfill- and increasing mucin secretion. Change in levels of serum ing inclusion and exclusion criteria were enrolled for the magnesium and CRP also has significant correlation with study & following tests were performed: Serum magne- asthma. sium estimated by Xylidyl blue with ATCS method, Serum

123 S80 Ind J Clin Biochem

COMPARISION OF ATYPICAL AND TYPICAL DRUGS IN SCHIZOPHRENIC PATIENTS FOR CAUSATION OF METABOLIC SYNDROME

Aditi Gupta, arun Raizada, taru Goyal, Dharamveer Yadav, Praveen Sharma Medanta- The Medicity Gurgaon, SMS Medical College, Jaipur

Schizophrenia is a devastating psychotic disorder and is kg and after 16 weeks of olanzapine treatment changed associated with metabolic syndrome after undergoing an- to 68.7+4.76 kg, it induces dyslipidemia (P<0.001) and hy- tipsychotic treatment. The study was designed to evaluate perglycemia (P<0.01). Clozapine was found to be second the impact of seven different antipsychotic in induction most potent drug in causation of metabolic syndrome as of metabolic syndrome in schizophrenic patients. A total the weight gain in clozapine treated patient was from the of 210 subjects (30 under each drug) with schizophrenia baseline 65.03+3.18kg to 67.83+3.53kg and with a significant were recruited according to ICD-10 criteria and were as- increase in glucemic and lipid parameters (P<0.001, P<0.01) signed to receive the drug for 16 weeks. Measurement of respectively. Aripriazole and amisulphride were compara- anthropometric (body weight, waist circumference, blood tively safer drugs as they were least potent in inducing pressure) and biochemical parameters (glucose, insulin, metabolic abnormalities (P>0.05) in schizophrenic patients HOMA-IR, HOMA-B, triglycerides, LDL, HDL) were done although their long term impacts need to be explored. As and the patients were subjected to ATP-III defined criteria clear from the study antipsychotics induces metabolic syn- for metabolic syndrome. The following outcome data were drome thus, the metabolic and cardiovascular risk factors obtained patients undergoing treatment with olanzapine should be surveillanced regularly in schizophrenic patients were highly associated with metabolic syndrome as it in- undergoing antipsychotic treatment. duces weight gain which at the baseline was 65.53+4.72

LEVELS OF UREA, CREATININE AND URIC ACID IN CHILDREN WITH SICKLE CELL DISEASE

Vikas K. Gupta*, Neelima Singh**, Prashant Nigam*, S.K.B. Patil* *Department of Biochemistry, CIMS, Bilaspur (C.G.), ** Department of Biochemistry, GRMC, Gwalior

Introduction: Uric acid is the end-product of purine me- healthy, age and sex matched controls were also chosen tabolism in man and is derived from the catabolism of for the study. Estimation of Urea, Creatinine, and Uric acid nucleic acids. Some uric acid is also produced during the done by standard kits methods. synthesis of nucleotides. Sickle cell disease (SCD) is clinically Result: The uric acid level was elevated in sickle cell pa- one of the most important haemoglobinopathies. Eryth- tients as compared with the normal control group. The rocytes containing mainly haemoglobin “S’ have a short value of Mean ±SD of the two groups: HbAA vs. HbAS life span. It would he expected that during erythropoisis was 5.19±0.64 // 9.4 ± 0.76 while for HbAA for HbSS was increased Synthesis of nucleic acid might occur, thus the 5.19±0.64 // 8.5±1.5, both being statistically highly signifi- destruction of red blood cells lead to increased nucleic cant. Urea and creatinine levels were considerably lower acid degradation. which means that lysis of red cells in in the sickle cell disease patients. The difference in the person with sickle cell disease does liberate the uric acid patient’s mean for urea compared to the mean in the content in the cell. Hyperuricemia was encountered in normal group (HbAA) was 32.37± 4.94 // 23±3.1 and 32.37± several studies on sickle cell disease patients. 4.94 // 23.8±2.84 for HbSS and HbAS, respectively. Likewise, Material and methods: A case – Control study carried out the difference in the mean for creatinine in HbSS group on 60 children subjects age 3 – 15 both male and female was 1.45±0.32 // 0.85±0.15 and in HbAS was 1.45±0.32 // Out of the 60 cases, 38 were heterozygous (20 males and 0.79±0.1, which was statistically significant. 18 females) and were 22 homozygous (12 males and 10 Conclusion: The results showed that the uric acid levels females). All cases of sickle cell anaemia were confirmed were increased while urea and creatinine levels were by haemoglobin electrophoresis and total number of 30 decreased in the sickle cell group patients as compared

123 Ind J Clin Biochem S81

with their age and sex-matched controls. Raised serum these patients as well as with other age groups for the uric levels were found in children with sickle cell disease. evaluation of their renal function. Clearance test studies are important to be carried out on

IDENTIFYING THE METABOLICALLY OBESE NORMAL WEIGHT MONW INDIVIDUALS IN SEDENTARY POPULATION

Shivani Jaswal, Jasbinder Kaur, Harjeet Kaur, Seema, Harpreet Kaur Walia Department of Biochemistry, Government Medical College & Hospital, Chandigarh

Aims & Objectives: The objective of this study was to Compared with individuals with BMI 18.5-20.9 kg/m2, the identify the metabolically obese normal weight individuals odds of metabolic syndrome were 1.33 (95% CI of 0.27-6.19) in a sedentary population and determine the likelihood of for BMI 21-22.9 kg/m2, 1.45 (95% CI of 0.35-6.0) for BMI metabolic syndrome in normal weight individuals. 23-24.9 kg/m2 and 3.40 (95% CI of 0.87-13.40) for BMI of 25-26.9 kg/m2. Material & Methods: The study was conducted by the Department of Biochemistry, GMCH, Chandigarh. The study Relative risk of developing metabolic syndrome was found group consisted of 250 volunteers who were leading a to be 4.58 (95% CI of 2.6-8.0) with increased TG, 3.19 sedentary lifestyle. Waist circumference, body weight, (95% CI of 2.2-4.5) with low HDL, 16.05 (95% CI of 3.6- height and BP of each participant were measured and 70.0) with increased FBS, 5.73 (95% CI of 3.31-9.9) with BMI was calculated. Fasting blood samples were obtained high BP and 18.3 (95% CI of 2.1-54.0) with increased waist and levels of Triglycerides, HDL and fasting blood sugar circumference. were estimated on HITACHI 902 analyzer by enzymatic Likelihood of metabolic syndrome was found to be sig- methods. Metabolic syndrome was defined according to nificantly increased with high TG, high FBS and increased ATP III guidelines. waist circumference. Low HDL and high BP however were Normal weight individuals (BMI < 25 kg/m2) were sub- found to have a moderate effect on likelihood of metabolic grouped according to BMI into three categories 18.5-20.9 syndrome in the otherwise normal weight individuals. kg/m2, 21-22.9 kg/m2, 23-24.9 kg/m2 and also compared Conclusion: Metabolically obese normal weight individuals with slightly overweight individuals (BMI 25-26.9 kg/m2). are those whose BMI was considered normal, but who had Results: The prevalence of metabolic syndrome in the nor- metabolic syndrome. It was observed that individuals in mal weight population was found to be 17.8% and increased the upper end and just above the normal BMI range are significantly with increase in BMI from 18.5kg/m2 to 26.9 more likely to have the metabolic syndrome as compared kg/m2. The prevalence of large waist circumference, high with those with BMI 18.5-20.9 kg/m2. The results of our TG, low HDL, high BP and high FBS increased significantly study suggest that screening for metabolic abnormalities (p<0.001) with increasing BMI. Large waist circumference should also be done for individuals at upper end of normal and elevated plasma fasting glucose concentration was weight spectrum, since the early detection of metabolically the least prevalent among normal weight individuals with obese normal weight individuals may be beneficial in the prevalence rates < 10%. prevention of diabetes and cardiovascular disease.

123 S82 Ind J Clin Biochem

CORRELATION OF SERUM FLUORIDE LEVELS AND OXIDATIVE BURDEN IN HYPERTENSIVE RETINOPATHY PATIENTS IN RAJASTHAN

Sudhanshu Mishra1, Anant Sharma2, Swati Tomar2, Manisha Choudhary1, Dushyant S. Chauhan1, Vivek Pratap Singh1, Sandeep Tripathi1, Anurag Tomar3 National Referral Centre for Fluoride Poisoning in India, 1Department of Biotechnology, NIET; Departments of 2Ophthalmology and 3Paediatrics, Nims Medical College. NIMS University, Shobha Nagar Jaipur Rajasthan, Jaipur

Background: Fluoride toxicity due to the presence of high (ROS), Malondialdehyde (MDA), protein carbonyl content levels of fluoride in our drinking water (>1.5ppm) may be (PC) and antioxidant namely superoxide dismutase (SOD), serious problems in our health. In Rajasthan, almost all catalase (CAT), glutathione peroxidase (GPx). In addition, districts have high Fluoride (up to 24.0 ppm) and about serum fluoride and lipid profile viz cholesterol, Triglyceride 11 million peoples are at risk. High fluoride ingestion has (TG), HDL, LDL and VLDL were determined in subjects. a definite relationship with the prevalence and severity of Results: Results of the study revealed that concentration diseases. There is very limited studies have been pointed of fluoride in serum was directly associated with increased out in association of hypertensive retinopathy (HRP) and cholesterol, TG, LDL, LPO and PC concentration while in- fluoride exposure. verse to the HDL and antioxidant levels in hypertensive Methodology: In the present study, we recruited 143 pa- patients as compared to positive controls. tients of HRP from the high fluoride region (>5.4 ppm) in Conclusion: It was concluded that hypertensive retinopathy Rajasthan and age and sex matched positive control (N= had higher fluoride content and in serum and it is strongly 149) were selected from the region where fluoride content associated with increased oxidative stress in hypertensive in water was less than 1.5 ppm. 5.0 ml of venous blood retinopathy patients. was collected for analysis of total reacting oxygen species

EVALUATION OF SERUM SODIUM, POTASSIUM, ALBUMIN AND CRP LEVEL IN PNEUMONIA

Shraddha Singh*, Dr. B.K. Agrawal*, Dr. Lokendra Dave**, Dr. Tripti Saxena*, Dr. Bhawna Bhimte* Department of Biochemistry*, Department of Medicine**, Gandhi Medical College Bhopal

Objective: To study the changes in serum sodium, potas- levels of serum CRP (p<0.001), Potassium (p<0.05), whereas sium, albumin and CRP level in pneumonia patients. significantly lower levels of serum albumin (p<0.001),

Methodology: A case control study was conducted on Sodium (p<0.001) as compared to normal healthy sub- 50 patients with pneumonia admitted to medical ward jects. at Hamidia hospital, Bhopal, and were compared with Conclusion: The decreased level of Na+ may be a marker 50 healthy individuals. Serum Na+ & K+ estimated by au- for severe underlying disease carrying a bad prognosis. toanalyser, serum albumin estimated by BCG method and The changes in level of serum potassium, albumin and serum CRP level estimated by Latex slide test. CRP to some extent could provide an illustration of the Result: Patients with pneumonia had significantly higher severity of infection.

123 Ind J Clin Biochem S83

STUDY ON MALNUTRITION, INFLAMMATION AND ATHEROSCLEROSIS MIA IN PREDIALYSIS PATIENTS

Shyam Krishnan. M*, Dr. Meera Shivashekar*, Dr. Padmanabhan**, Dr. Ebenezer William* Department of Biochemistry and Department of Nephrology, SRM Medical College Hospital & Research Centre

Objective: The study was performed to determine any trols showed a significant increase in Inflammatory and strong association between Malnutrition, Inflammation and atherosclerotic markers. In predialysis patients the levels Atherosclerosis (MIA) in Predialysis patients. of Ferritin, Transferrin and hsCRP was found to be signifi- cantly increased than the controls. But Albumin levels are Method: The study was carried out in 45 Hemodialysis decreased significantly in predialysis group. patients attending the Nephrology unit of SRM Medical College Hospital and Research Centre. 30 healthy individu- Conclusion: The study suggests a strong association als were included as controls in this study. Blood samples between the markers of Malnutrition, Inflammation and were collected from both groups and markers of MIA Atherosclerosis (MIA) in predialysis patients. This study were assessed. suggests the presence of MIA syndrome which is associ- ated with an exceptionally high mortality rate. Results: The study including 45 patients on regular he- modialysis (21 yrs and above) and 30 sex matched con-

PROTECTIVE ROLE OF DOCOSAHEXAENOIC ACID ON LITHIUM INDUCED CEREBELLAR TOXICITY AND ASSOCIATED NEUROMUSCULAR STATUS IN MALE ALBINO RATS

Mehta Tathagat, Ahmad Nadeem, Manisha Choudhary, Sudhanshu Mishra, Dushyant Singh Chauhan, Sandeep Tripathi Department of Biotechnology, NIET, Nims University, Jaipur

The present study attempts to assess the protective ef- significant changes in behavioral profiles but these changes ficacy of docosahexaenoic acid (100 mg/kg body weight) were reversed to near control following the treatment on lithium chloride (100 mg / kg b. wt.) mediated oxida- of DHA. The light microscopic and ultrastructural studies tive damage in the cerebellum of male albino rats (N=10) revealed damaged Purkinje’s neurons and altered granu- along with the associated dysfunctioning of neuromuscular lar cell layer along with the increased accumulation of coordination and motor activity. A significant decrease in lipofuscin granules in lithium treated rats. These changes the activities of antioxidant enzymes and increased total were quite less pronounced in DHA treated group than reacting oxygen species, lipid and protein peroxidation that of lithium and this may be due to the reduction of products observed in lithium exposed rats. We observed oxidative burden by DHA. On the basis of our results it that treatment with DHA restored the altered antioxidant may be concluded that excessive lithium may be linked enzyme activities, when compared with lithium treated with cerebellar degeneration and neuromuscular disorders rats. Moreover, lithium treated groups of rats exhibited while DHA ameliorating these effects.

EFFECT OF POLYPHENOLIC ACETATES ON NFКB GENE EXPRESSION IN LUNG CANCER

Anju Sharma1,2, Vishwajeet Rohil2, Kapili Nath2, Shweta Paul2, Jayashree Bhattacharjee1 1Vardhman Mahavir Medical College & Safdarjung Hospital, New-Delhi, 2Department of Clinical Biochemistry, Vallabhbhai Patel Chest, Delhi

Objectives: Lung cancer has been the most common cancer all the new cancers. Alteration of genes occurs in cancer in the world for several decades, and by 2008, there were either by mutation or epigenetics. Polyphenolic acetates an estimated 1.61 million new cases, representing 12.7% of (DAMC* and EAA**) have been shown earlier to induce

123 S84 Ind J Clin Biochem

Apoptosis and cell cycle arrest in lung cancer. NF-kB is a protocol. This cDNA was used as a template for NF-КB gene transcription factor that controls the expression of genes quantification using suitable primers by Real-time PCR. involved in immune responses, apoptosis and cell cycle and Results: The data was analyzed using Real-time PCR Soft- it has been implicated in several malignancies. Here we ware from Applied Biosystem and there is a statistically are analyzing that induction of Apoptosis in lung cancer significant increase in the expression of NF-КB gene as is NF-КB dependent or not. obtained by the Ct value. a. To treat lung cancer cells (A549) with various combina- tion of Polyphenolic Acetates. Conclusions: Polyphenolic Acetates have shown to sig- b. To study the effect of Polyphenolic acetates on lung nificantly increase apoptosis in a NF-КB dependent in cancer cells (A549) by analyzing the change in NF-КB lung cancer cells. This study can further help in better gene expression by Real time PCR. understanding of the underlying mechanism that lead to apoptosis in lung cancer. So, it might provide a new insight Methodology: Non small lung cancer cells (A549) are to the role of polyphenolic acetates as chemopreventive treated with various Polyphenolic acetates (DAMC* and drugs for lung cancer treatment. EAA**). Total cellular RNA was extracted and reverse transcribed using Reverse transcriptase enzyme (Moloney Keywords: DAMC* = 7,8 -Diacetoxy-4-Methyl Coumarin and murine leukemia virus) according to the manufacturer’s EAA** = Ellagic acid peracetate

HYPOCHOLESTEROLEMIC AND ANTIOXIDANT EFFECT OF METHANOLIC EXTRACT OF MUSA SAPIENTUM LINN

Rimi Shukla, Piyush Dikshit, Mool K Tyagi, Kirikar Shukla, Jaswinder K Gambhir Corresponding Author:-Dr. Rimi Shukla, Professor, Room no.217, Department of Biochemistry, University College of Medical Sciences and Guru Teg Bahadur Hospital, Dilshad Garden, Delhi

Objective: Hypercholesterolemia contributes significantly Results: Six weeks treatment of hypercholesterolemic rats in the prevalence & severity of atherosclerosis and coro- with methanolic extract decreased total cholesterol by 47%, nary heart disease. Medicinal plants are measure source LDL+ VLDL-C by 62% and TAG by 30%. HDL-C increased by of drug in Indian and other systems of medicine. Present 21%. Atherogenic index (LDL + VLDL-C/HDL-C) decreased study was planned to assess antihypercholesterolemic & significantly (p<0.01). Treatment with Msmt decreased antioxidant effect of methanolic extract of stem of Musa MDA, a marker of lipid peroxidation, in blood as well as sapientum – a herbaceous plant of musaceae family, in liver tissue whereas reduced glutathione and antioxidant hypercholesterolemic rats. enzymes, superoxide dismutase and catalase increased significantly (p<0.01). Methods: Rats were made hypercholesterolemic by feeding cholesterol (100mg/kg). Methanolic extract (20mg/kg) was Conclusion: Methanolic extract of stem of Musa sapientum administered along with cholesterol suspended in soya oil has significant antihypercholesterolemic & antioxidant ef- to rats once daily orally for 45 days. Cholesterol, Triacylg- fect. It has potential to be used for development of drug lycerol & HDL-C was measured on 0 and 45th day. At the for treatment of hypercholesterolemia. end of the experiment, antioxidant enzymes (superoxide dismutase and catalase), reduced glutathione and catalase were determined in blood and liver tissue.

123 Ind J Clin Biochem S85

EVALUATION OF OXIDATIVE STRESS IN OBESE MEDICAL STUDENTS WITH REFERENCE TO CARDIOMETABOLIC SYNDROME

Meghana K Padwal1, Priya Kamthe2, R.R. Melinkeri3 1Associate Professor, 2Post Graduate Student, 3Professor and Head, Department of Biochemistry, BVDUMC Pune

Background: The prevalence of obesity in young population levels were done by automated biochemistry analyser. is on rise. The major complication of obesity is dylipedemia, Estimation of hs-CRP was done by automated chemilumi- impaired glucose metabolism etc. It has been demon- nescence immunoassay system. strated that fat accumulation correlates with markers of Results: There was statistically significant rise in the levels systemic oxidative stress. Oxidative stress may have pro- of hs-CRP and MDA in obese students as compared to non inflammatory role on promoting atherosclerosis. obese (p=0.001). There was no statistically significant dif- Objectives: The present study was designed to evaluate ference in the levels of blood glucose in both the groups the relationship between the marker oxidative stress i.e. (p = 0.06). However the levels of Total cholesterol, LDL Malondialdehyde (MDA) with reference to markers of were significantly increased in cases (p = 0.01), while Cardiometabolic Syndrome i.e. hs- CRP, lipid profile and levels of HDL were significantly decreased (p = 0.01) as blood glucose in healthy obese students. compare to controls.

Material and Methods: The present study was a prospec- Conclusions: There is increased oxidative stress in obesity tive case control study, carried out in a university medical which may aggrevate the proinflammatory processes of college. The participants were healthy medical students of atherosclerosis. The metabolic changes in obesity are asso- both the genders. They were divided into cases: Obese ciated with to chronic systemic low grade inflammation. (BMI ≥ 25 kg/m2; n=30) and controls: Non obese (BMI ≤ Keywords: Oxidative stress, Cardiometabolic Syndrome, 25 kg/m2; n=30). Estimation of MDA, Lipid Profile, glucose MDA, hs – CRP.

COMPARATIVE STUDY OF METABOLIC CHANGES OCCURS IN DIFFERENT MALIGNANCIES

Varma M1, Paneri S2, Singh J.D.3, Yadav S4, Ghanghoria S5, Paneri L6 1Dept. of Biochemistry Shree Arbindo institute of medical sciences. 2,3Department of Biochemistry M.G.M. Medical college Indore, 4Dept. of Gynecology M.G.M. Medical college Indore. 5Dept. of Pathology M.G.M. Medical college

Cancer presenting clinically with number of metabolic B-cell lymphoma, t-cell lymphoma) and compared it with changes which are characterized by irreversible morpho- healthy control subjects by estimating serum biochemical logical and functional alterations in corresponding areas. parameters like glucose, urea, creatinine, uric acid, al- The present study was undertaken in department of kaline phasphatase, SGOT, SGPT, Total protein, albumin, biochemistry MDA. The serum levels of above biochemical parameters were significantly changed in study patients compared to from 1990 to 2011 and study comprised 500 patients suf- control group (p<0.001). However there was significant fering from different types of malignancies. The objectives difference in serum MDA levels (p< 0.05) between dif- of the present study was to investigate the patterns of ferent malignancies. Among all malignancies NHL T-cell metabolic changes in patients suffering from different ma- lymphoma patient shows highly significant alteration in lignancies (breast cancer, cancer cervix, prostate cancer, their biochemical parameters.

123 S86 Ind J Clin Biochem

ALTERATIONS IN THE LEVELS OF PLASMA AMINO ACIDS IN POLYCYSTIC OVARIAN SYNDROME

Sumithra. N. Unni. C*, Kannan Vaidyanathan*, Lakshmi. R. Lakshman**, Leela Menon* *Dept. of Biochemistry, **Dept. of Obstetrics and Gynecology, Amrita Institute of Medical Sciences, AIMS Ponekkara PO, Kochi

Background: Plasma amino acid levels are altered during HPLC-Shimadzu; Column- LUNA C18, Detection-Absorption stress. Polycystic ovarian syndrome is a condition which (254nm). Samples were deproteinised and treated with is associated with metabolic stress. This study was under- Phenyl isothiocyanate (PITC) and triethylamine (TEA) prior taken to find out the level of amino acids in Polycystic to injection (precolumn derivatization). ovarian syndrome. Results: The levels of Methionine, Cystine, Isoleucine, Phe- Aim: To analyse the levels of the following amino acids nylalanine, Valine, Tyrosine, Proline, Glycine (all p<0.001), in plasma in Polycystic ovarian syndrome: Glycine, Ala- Lysine (p=0.001) and Histidine (p=0.016) were found to be nine, Valine, Leucine, Isoleucine, Tyrosine, Phenylalanine, significantly lower in cases than controls. Arginine (p<0.001) Methionine, Cystine, Serine, Histidine, Arginine, Lysine and Alanine (p=0.001) were found to be significantly higher and Proline. in cases. Level of Serine (p=0.064) was found to be higher and statistically significant; but in the borderline. Materials and methods: 65 female patients in the age group 20-40 years were included in this study. Amino ac- Conclusion: Significant derangement in the levels of plasma ids levels were determined using High Performance Liquid amino acids are found in Polycystic ovarian syndrome. Chromatography. Analytical conditions were as follows:

UNCONTROLLED HYPERGLYCEMIA: A PROBABLE CAUSE OF INFLAMMATION AND OXIDATIVE STRESS IN TYPE 2 DIABETIC NEPHROPATHY SUBJECTS

Deepak Kafle, Neelima Singh, S.K. Singh, Nivedita Singh, Vishal Bhargav and A. K. Singh Department of Biochemistry, GR Medical College, Gwalior

Background: Oxidative stress increased in diabetes gener- (P<0.01), with group 2 having more significant than group ates ROS producing inflammatory cytokines tumor necrosis 1 (P<0.01). Antioxidant enzyme glutathione peroxidase was factor (TNF)-α and interleukin (IL-6) in renal cells, which found to be decreased in both groups of diabetic patients are the factors responsible for diabetic nephropathy. as compared to controls, (P<<0.05), with group 2 having The study aimed to predict the development of diabetic significantly decreased as compared to group 1 (P<0.05). nephropathy in type 2 diabetes due to inflammation and Glutathione reductase was also found decreased in group oxidative stress via uncontrolled hyperglycemia.. 2 subjects as compared with group 1 and control subjects. Serum urea and creatinine levels were increased in group Methods: Serum levels of inflammatory markers (IL-6 2, (P<0.05), but were within normal range in group 1 and and Tnf-α), antioxidants, [Glutathione reductase (GR) and healthy controls. Glutathione peroxidase (GPx)], plasma malondialdehyde (MDA), fasting blood sugar, urea and creatinine levels Conclusion: Results of the present study indicates that were estimated in controls (n=25), diabetes without dia- inflammatory markers are increased with decreased antioxi- betic nephropathy (n=25, group1) and with nephropathy dant defense levels in patients with diabetic nephropathy (n=50, group2). due to uncontrolled hyperglycemia.

Results: Serum levels of inflammatory markers (IL-6 and Keywords: Oxidative stress, inflammation, diabetic neph- Tnf-α), malondialdehyde, fasting blood sugar, were high in ropathy. both groups of diabetic patients as compared to controls,

123 Ind J Clin Biochem S87

ASSOCIATION BETWEEN POSTPRANDIAL DYSLIPIDEMIA IN TYPE II DIABETIC SUBJECTS AND URINARY ALBUMIN EXCRETION A CASECONTROL STUDY IN NORTH INDIAN POPULATION

Sarika Arora, Prabhat K. Singh, Rulie Buragohain, Shakti Aggarwal, R.K. Kapoor Department of Biochemistry, ESI Postgraduate Institute of Medical Sciences & Research, Basaidarapur, New Delhi

Background: Postprandial dyslipidemia and microalbumin Result: Fasting and postprandial blood sugar levels and excretion have been independently shown to increase insulin levels were significantly higher in diabetic subjects cardiovascular disease risk in Type II Diabetic subjects. How- as compared to the control group (p<0.01). The difference ever, the association between postprandial dyslipidemia in triglyceride levels was not found to be significant in and microalbumin excretion has not been studied. fasting state (p=0.051) but in postprandial state highly significant difference was observed between the diabetic Method: This case-control study included 105 subjects with subjects and the control group (p=0.001). Serum triglycer- Type II Diabetes Mellitus and 64 age and sex matched ide levels in diabetic subjects showed a significant positive healthy controls. Fasting and 2-hour postprandial blood correlation with urinary albumin excretion in these subjects samples were collected from all the subjects and analy- (r= 0.444, p<0.001). sed for plasma glucose, serum insulin, serum cholesterol, triglycerides and high-density lipoproteins. In addition Conclusion: Diabetic subjects excreting microalbumin in fasting samples were analysed for Kidney function tests urine have exaggerated postprandial lipemia than control (Serum urea, creatinine and uric acid levels) to rule out subjects and diabetic patients with no microalbumin ex- overt Renal disease and glycated haemoglobin levels. A cretion. Microalbuminuria in patients with diabetes might spot urine sample was collected from the patient within reflect widespread vascular damage resulting in low Li- the two hours waiting period and analyzed for urine mi- poprotein lipase activity and hence delayed clearance of croalbumin/ creatinine ratio. triglyceride rich lipoproteins from plasma.

ESTIMATION OF THE LEVELS OF LIPID PEROXIDATION AND TOTAL ANTIOXIDANT CAPACITY IN HYPERTHYROID, HYPOTHYROID & EUTHYROID SUBJECTS A CROSSSECTIONAL STUDY

Dr Vanishree B J1, Dr Anil M2, Mr. Basant Joshi2, Dr Smita S Sonoli3, Mr Mohit3 Miss Deepali3 1,2,3Department of Biochemistry, J. N. Medical College, Belgaum, Karnataka India

Variation in thyroid hormones (TH) is the most important Mean±SD, statistical analysis performed using one way factors involved in the regulation of the basal metabolic ANOVA considering p<0.05 as lowest limit of significance. condition as well as in the oxidative metabolism. TH All these parameters differed significantly among the stimulates the production of free radicals and their dis- groups. The level of MDA was significantly higher in cases orders have pathogenic impact on human tissues. MDA, of hyperthyroid when compared with euthyroid subjects. breakdown product of major chain reaction leads to No change was observed in hypothyroid when compared definite oxidation of PUFA & serves as reliable marker of with normal subjects. The levels of TAC decreased sig- lipid per-oxidation (LPO). Total Antioxidant Capacity (TAC), nificantly in both hyperthyroid and hypothyroid patients instead of individual antioxidants, and LPO with changes when compared with euthyroid subjects. In conclusion, in TH were seen in this study. Serum T3, T4 & TSH by hyperthyroidism may cause LPO in tissues whereas hypo- chemiluminescence assay, LPO by Okawa H et al (modi- thyroidism has no effects on LPO. The hyperthyroidism and fied) and TAC by D Koracevic et al were estimated in 40 hypothyroidism may cause changes in antioxidant defence eu-thyroids, 20 hyperthyroids and 30 hypothyroids subjects system indicating increased tissue damage with alteration of age group of 25-50 years. The data was expressed as of thyroid hormones.

123 S88 Ind J Clin Biochem

PROTECTIVE ROLE OF VISFATIN IN NON ALCOHOLIC FATTY LIVER DISEASE: A PROOF OF CONCEPT, EXPERIMENTAL ANIMAL STUDY

Sadik Ali Mahammad1, R M Mukarjee1, P N Rao2, D N Reddy2, M Sasikala1 Institute of Basic Science and Translational Research1, Asian institute of Gastroenterology2, Hyderabad

Introduction: Visfatin/PBEF is a protein (56 kDa) secreted Visfatin (fed with HFD and received Intraperitonial injection by visceral adipose tissue with which metabolic disorders of Recombinant Rat Visfatin daily for 3 weeks). After 3 in humans have been associated. Visfatin was found to weeks, the rats were sacrificed to collect the liver tissue have anti apoptotic activity and regulatory role in inflam- and blood for biochemical parameters. mation. Previous study demonstrated association between Results: significant (p<0.05) increase in rat body eight, decreased visceral adipose tissue Visfatin and degree of ste- liver weight, ALT, cholesterol, triglycerides and NEFA were atosis in NAFLD patients (Sasikala et al, Ann Hepato1.2010 noted in HFD group (Group2) compared to controls. Vis- Jul-Sep;9(3):266-70).•:•in this study our objective is to fatin treatment (Group 3) significantly reduced the body evaluate its protective role on liver in high fat fed rats. weight, liver weight, ALT, triglycerides, LDL and NEFA in Materials and Methods: Animals:-Male Sprague-Dawley animals kept on HFD (Group 2). Rats weighing 180 10 gm and aged 7 to 8 weeks, Ani- Conclusion: The findings of this study shows decreased ALT mals were randomly divided into 3 groups.) Group 1 (n=5) levels in High Fat Diet fed rats treated with Visfatin. This Control (fed with Normal Rat diet) DGroup2 (n=5) High suggest a Putative Protective role of Visfatin in Non Alco- Fat Diet (fed with High Fat Diet) >Group 3 (n=7) HFD + holic Fatty Liver Disease in Experimental animal study.

STUDY OF LIPID PROFILE IN HIV POSITIVE PATIENTS

Asha kumari

With increase in life span, HIV positive subjects are fac- low density lipoprotein cholesterol (VLDL-C) & Triglycerides ing more cardiovascular complications, because of HIV (TG) were taken. CD4+ cell counts were assessed in HIV infection and ART. Many studies have shown that higher positive subjects & three groups were made as follows: CD4+ count was associated with increased HDL-Cholesterol Group1 (CD4+ counts <200cells/ μL, n=52), Group 2 (CD4+ while higher TG and VLDL-Cholesterol levels were seen at counts 200-350cell/μl, n=18), Group 3 (CD4+ counts >350cell lower CD4+ count. / μL, n=54).

Aim and Ojectives: Results: Mean of TG in Group 1, 2 and 3 were 194.4mg/dl, 1. To assess and compare lipid profile in HIV positive 139.5mg/dl and 159.5mg/dl. Mean of TG and VLDL in HIV subjects with controls. patients were 167.9mg/dl & 33.5mg/dl, significantly higher 2. To find correlations if any with their CD4+ cell (p<0.05) than that of control group (TG=94.2mg/dl and counts. VLDL=18.8mg/dl). HDL in 53.41% HIV positive subjects were below 30mg/dl. Rest parameters were not significantly Materials and Methods: 113 HIV positive subjects attending different between cases and controls. ART centre of Pt BDS PGIMS, Rohtak and 31 age matched controls were taken for study. Fasting serum levels for Conclusion: Lipid profile should be evaluated in every pa- total cholesterol (TC), high density lipoprotein cholesterol tient of HIV infection, to detect derangements which can (HDL-C), low density lipoprotein cholesterol (LDL-C), very lead to earlier cardiovascular complications.

123 Ind J Clin Biochem S89

STUDIES ON REACTIVE OXYGEN SPECIES AND DNA DAMAGE IN EXPERIMENTAL DIABETES

Jayant Shiv Kumar, Srivastava Nalini SOS in Biochemistry Jiwaji University Gwalior

Diabetes is a chronic endocrine disorder, which occurs drate to overnight fasted rats. Rats were sacrificed after when the pancreas does not produce enough insulin, this 48h and the levels of hydrogen peroxide and DNA single lead to an increased concentration of glucose in the blood strand breaks were measured. The results suggested that (hyperglycemia). There are several secondary complications the levels of hydrogen peroxide were increased in all rats reported in diabetes. Present study was aimed to evaluate tissues which were given alloxan. The levels of DNA dam- the level of reactive oxygen species and consequent DNA age was also increased in rat tissues which have blood damage in rat tissues by single cell gel electrophoresis glucose levels above 300 mg/dl. The results clearly show (comet assay), in experimental diabetes. The rats of Wistar that experimental diabetes caused increase in the levels strain were selected as animal model and diabetes was reactive oxygen species and DNA damage in rat tissues. induced by single intravenous injection of alloxan monohy-

CONFIRMING AND MONITORING FORMS OF PROSTATE CANCER BY SELECTIVE TUMOR MARKERS PSA OR PAP?

V Mehrotra and G Awasthi Biochemistry Department, Himalayan Institute of Medical Sciences, Jollygrant and *Dolphin Institute of Para Medical Sciences, Dehradun

Background: The past decade has witnessed a marked by elevation of serum PSA levels in men with prostate increase in the interest in prostate cancer, reflecting nodular hyperplasia. the desire for improvements in treatment of all forms Method: To evaluate possible discrimination of prostate of cancer. Prostate cancer is the most common visceral adenocarcinoma from prostate nodular hyperplasia, the cancer with second leading cause of cancer deaths and serum PSA and PAP concentrations were measured in 50 major cause of morbidity and health care expenditure. patients with prostate adenocarcinoma and 50 patients Both the incidence and age-adjusted mortality rates of with nodular hyperplasia. prostate cancer have increased substantially in resent years. Despite significant advances in the diagnosis, staging Results and Conclusions: 95% of patients with prostate and treatment the optimal management of this disease carcinoma having serum PSA level >4 ng/ml indicate higher remains undefined. sensitivity for detecting prostate cancer, however 36% pa- tients with prostate hyperplasia has PSA levels >4 ng/ml. Prostatic specific antigen (PSA) and prostatic acid phos- Results indicate that serum PSA can discriminate prostate phatase (PAP) are the tumor markers used for confirming adenocarcinoma from Prostate nodular hyperplasia better and monitoring prostate cancer. The clinical utility of PSA than PAP. and PAP for early detection of prostate cancer is hampered

OXIDANT & ANTIOXIDANT STATUS IN CEREBROSPINAL FLUID IN CHILDREN WITH SEPTIC MENINGITIS

Ragini Srivastava1, Rajiv Lohokare1, Rajniti Prasad2 1Department of Biochemistry, 2Department of Pediatrics, Institute of Medical Sciences, Banaras Hindu University, Varanasi

Aim: Septic meningitis is the most severe and frequent in- sequelae in 50% of survivors after bacterial meningitis. The fection of the central nervous system (CNS). Several studies mechanisms of CNS damage during meningitis have not confirmed that brain damage is responsible for neurological been conclusively identified. During bacterial infections,

123 S90 Ind J Clin Biochem

neutrophils and macrophages gather at the site of infec- ene vials stored at -20˚C for analysis of malondialdehyde, tion to combat the microorganisms. Levels of antioxidant protein carbonyl, nitrite, superoxide dismutase, ascorbic and oxidant become imbalanced during bacterial meningitis acid, glutathione peroxidase till estimation. and produce free radicals and oxidative stress. The rela- Result: The CSF MDA, protein carbonyl and nitrite levels tionship between oxidative stress/antioxidant mechanisms were significantly raised in the cases of septic meningitis as in patients with bacterial meningitis is barely determined. compared to controls (p <0.001). The plasma glutathione, The present study has been taken up to know the levels SOD and ascorbic acid level were significantly decreased of different markers of oxidative injury and antioxidant in cases with septic meningitis as compared to controls capacity in septic meningitis. (p <0.001). Methods: The present study included 40 children with Conclusion: Oxidative stress may be associated with the newly diagnosed cases of meningitis and 40 children pathophysiology of meningitis and that its clinical attenu- matched for age and sex were taken as controls. After ation may lead to improvement in the prognosis for the having informed consent, under aseptic precaution cere- disease. brospinal fluid (CSF) was collected in deionized polyethyl-

ESTIMATION OF NON CONVENTIONAL RISK FACTOR FOR CARDIAC RISK EVALUATION IN RURAL AND URBAN POPULATION

G. Awasthi1, S. Kumar1 Lokendra1 Vinit Mehrotra2 and M.S.M. Rawat3 1Department of Biochemistry. Dolphin (PG) Institute of Bio-medical and Natural Science. Dehradun, 2Department of Biochemistry. Himalyan Institute of medical Science Jolly Grant Dehradun, 3Department of Chemistry, H.N.B. Garhwal University, Srinagar Garhwal

Background: Lp[a] is lipoprotein particle found in the volunteers. Age, height, weight and blood pressure were blood stream and is a major independent risk factor for recorded and food habits of the subjects were obtained myocardial infarction. Approximately 30% of individuals with using a questionnaire. The past history of persons along heart disease have elevated Lp[a] levels. Fibrinogen is con- with family history was tracked with no cases of M.I. Lp[a], verted to fibrin during the clot formation and is reported Fibrinogen and Homocysteine was estimated. as newer risk factor for MI. Homocysteine increases the Result: No significant difference in Lp[a] levels was pathogencity of LDL. found between the two population. The fibrinogen and The present study was aimed at estimating LP [a] levels, Homocysteine levels were found to be greater in urban Fibrinogen and Homocysteine in rural and urban popula- as compared to rural population. Greater level of Homo- tion of Dehradun region. cysteine and fibrinogen in urban population as compared to rural population found in the present study put the Method: A total of 56 (Males= 28 and Females= 28) in- urban population to the greater risk of developing coro- dividuals aged between 20- 60 years were selected from nary heart disease. rural and urban population. 5 ml intravenous blood was drawn from each individual in resting position after over- night fasting. An informed consent was taken from the

PREVALENCE OF GLUCOSE6PHOSPHATE DEHYDROGENASE DEFICIENCY IN TERM BABIES WITH NEONATAL HYPERBILIRUBINEMIA IN A TERTIARY CARE HOSPITAL

Monika Chadha Objective: Glucose-6-phosphate dehydrogenase (G6PD) the cause of neonatal jaundice. Our aim was to study the deficiency is the most prevalent enzyme deficiency world- prevalence of G6PD deficiency in neonatal hyperbilirubine- wide. The prevalence of G6PD deficiency ranges from 0.2% mia in a tertiary care centre. to 19.0% in India. It is an inherited deficiency that may be

123 Ind J Clin Biochem S91

Material and Methods: This retrospective study was con- not found associated with AB+ve blood group and all the ducted amongst neonates with hyperbilirubinemia admitted Negative blood groups. to Neonatal Intensive Care Unit of Satguru Partap Singh Conclusion: In this study, the prevalence of G6PD defi- Apollo Hospital, Ludhiana from January 2011 to August ciency in neonatal hyperbilirubinemia was considerably low. 2012. The dataset included: Age, Sex, Total Bilirubin, Blood Yet the importance of identifying G6PD deficiency in the Group and Rh of mother and newborn, Direct Coomb’s newborn period cannot be overlooked so that appropriate test and G6PD level. counseling is provided to the family and potentially harm- Results: Out of 224 noenates with bilirubin level more than ful drugs are not administered to these infants and their 10 mg/dL on phototherapy, 5 neonates (2.23%) were found nursing mothers. Thus, we recommend that a screening to have G6PD deficiency. The male to female ratio was 4: program should be introduced to cover all the neonates 1. Rh and ABO incompatibility were not seen in any of the for identification of G6PD deficiency in all institutional 5 patients with G6PD deficiency. The mean bilirubin level deliveries at tertiary care hospitals. of the study group was 18.3 mg/dL. G6PD deficiency was

ENDOGENOUS SEX HORMONES AND GLYCEMIC CONTROL IN POSTMENOPAUSAL WOMEN HAVING DIABETES TYPE 2 IN NORTH INDIA

Yadav P, Seth S, Chugh K, Gehlot V, Sachdev A

Introduction: In postmenopausal women, endogenous measured on Autoanalyser via Immunoassay Kits. The estrogen and testosterone have been positively associ- results were analysed and compared. ated with glucose intolerance and type 2 diabetes. Most Results: The mean age was 64.7 years (SD ± 8.7) and studies have not examined these associations in a large n=1250. HbA1c was significantly higher (p<0.05) in women group of postmenopausal women in India. having low Estrogen as compared to women having normal Objective: To study the effect of Endogenous Sex Hor- Estrogen levels (age-adjusted mean ± SE: 8.5 ± 0.02 vs. mones on Glycemic Control in postmenopausal women 6.6 ± 0.03, respectively). HbA1c levels were significantly having Diabetes type 2 in North India. higher (p<0.05) in women with high Testosterone levels as compared to women having normal Testosterone lev- Material And Methods: Only diagnosed cases of diabetes els (age adjusted mean ± SE: 8.0 ± 0.02 vs. 6.9 ± 0.03, type 2 (n=1250 women) aged >55 years years undergoing respectively). glucose profile testing were included. Serum testosterone (normal=300-1000 ng/dl), serum estrogen in post-menopaus- Conclusion: Overall results clearly demonstrate that Low al women (upper limit-35 pg/ml) and HbA1c (normal=4-5.6% Estrogen levels and High Testosterone levels in North In- in normal people, <6.5% -target control in diabetics) were dian Postmenopausal Females having Diabetes type 2 are associated with Poor Glycemic control.

EFFECT OF OXYTETRACYCLINE ON IN VITRO MINERALIZATION AND DEMINERALIZATION REACTIONS IN THE ABSENCE AND PRESENCE OF COLLAGEN

Monica Kakkar*, Ashok Kumar Lal*, Vineet Mehrotra*, Raj Kumar Jethi** *Himalayan Institute of Medical Sciences, Dehradun **Punjab University, Chandigarh

Background: Oxytetracycline and its derivatives routinely as bone fluorochrome to study bone metabolism. However, used to treat various ailments have also been shown to in- despite the above observations, its mechanism of action hibit embryonic bone formation, mineralization in pregnant is not clearly understood. Controversy exists whether or female rats and parathyroid hormone induced demineraliza- not it acts by influencing collagen biosynthesis. tion of bones. Oxytetracycline has also been routinely used

123 S92 Ind J Clin Biochem

Objective: To study the mechanism by which Oxytetracy- collagen) oxytetracycline was found to inhibit both the clin influences in vitro mineralization and demineralization initial mineral phase formation and its subsequent growth reactions in the absence and presence of collagen without influencing the demineralization. However, when in the heterogeneous system mineralization was studied Materials and Methods: Homogeneous and Heterogeneous in the presence of collagen, oxytetracycline was found systems of In vitro mineralization under physiological condi- to inhibit not only the initial mineralization but also the tions were used to investigate the effect of oxytetracyline subsequent growth and demineralization of the preformed not only on initial mineralization but also the subsequent mineral phase. growth and demineralization of the preformed mineral phase both in the absence and presence of collagen. Conclusions: Oxytetracycline acted like crystal poisons to inhibit the mineralization and demineralization reaction Results: The studies demonstrated that in the homoge- reactions by tightly associating with the mineral phase. neous reaction system (mineralization in the absence of

STUDY OF GLYCOSYLATED HAEMOGLOBIN HBA1C IN TYPE2 DIABETES MELLITUS PATIENTS IN AGRA POPULATION

K P Mishra*, Alok Mawar, Pawan Kare and Manju Kumari Deparment of Biochemistry, S N Medical College, Agra

The present Study on “glycosylated haemoglobin (HbA1C) higher found in diabetic male as compared to that of among type 2 diabetes mellitus patients in Agra popula- healthy subjects. No significant change in FBS and HbA1c tion” was conducted. For this, 40 clinically established levels was observed in diabetic male patients as compared hyperglycaemic patients suffering from type-2 diabetes to that found in diabetic female patients. The increase in mellitus, attending the O.P.D. of Department of Medicine, FBS level was associated with increase in HbA1c levels. S.N. Medical Collage & Hospital, Agra were selected under FBS levels increased significantly with the increasing dura- study. The result was compared with 50 age matched tion of diabetes. Glycosylated haemoglobin level increased healthy, non-obese, non diabetic control subjects. The significantly with the increase in duration of diabetes. The aim of study was to find out relationship between HbA1c increasing duration of diabetes also influenced the glycemic and duration of diabetes mellitus. Significant increased control in diabetic patients. Therefore, a regular monitoring in fasting blood sugar (FBS) and HbA1c levels in diabetic of HbA1c in diabetic patients with increasing duration of patients were observed when compared to that of healthy diabetes is helpful to assess the further complications. controls. Similarly, FBS and HbA1c levels were significantly

ELEVATED EXPRESSION LEVEL OF SURVIVIN PROTEIN IN EARLY STAGE OF HUMAN GASTRIC CANCER

Anbarasu kannan, Niranjali Devaraj*, S. Subaramaniam**, Shyama Subaramaniam Department Biochemistry, Apollo Hospitals, Chennai. *Department Biochemistry, University of Madras, Chennai, **Regenix super speciality laboratories, Chennai.

Objective: To identify the role of survivin in early gastric at the G2-M phase. It is a nuclear shuttle protein that is cancer. To study the relationship between survivin and actively exported from the nucleus. histological differentiation of grades of gastric cancer. Methdology: Immunohisto staining of survivin by using Introduction: Survivin is a member of the inhibitor of the monoclonal antibody was performed by the standard apoptotic protein family, represents a multifunctional pro- streptavidin-peroxidase (SP) technique. The survivin pro- tein that suppresses apoptosis and regulates cell division tein level was quantified by a ELISA and by western blot

123 Ind J Clin Biochem S93

analysis of 45 early gastric cancer patient and 10 normal western blot, we found a significant correlation in both individuals. methods. However in a direct comparison of both survivin protein detection assays, we found a higher sensitivity and Results: Expression of survivin was observed in the cyto- stronger correlation of prognosis in survivin ELISA. In nor- plasm and nucleus in pathological grade of early stage. mal individuals expression of survivin is not observed. Survivin proteins were expressed in 38 of the 45 (80.59%) patients. High survivin protein levels were detected by Conclusion: The survivin expression is an essential event ELISA and western blot analysis in tumour tissue. When in the early stage of gastric cancer. It may thus be a new compare with survivin values obtained by ELISA (or) diagnostic and therapeutic target in early gastric cancer.

MOLECULAR FINGERPRINTING OF S. MAXIMA AND EVALUATION OF ITS ANTI HYPERGLYCEMIC ACTIVITY IN STREPTOZOTACIN INDUCED DIABETES

Anish Zacharia. J, Senthil Kumar S, Urmila J, Prasad GBKS, and Bisen PS. SOS in Biochemistry, Jiwaji University, Gwalior, Defence Research & Development Establishment, Gwalior

Spirulina maxima is a filamentous cyanobacterium well Results: Therapy with aqueous and ethanolic extracts of known for its neutraceutical activity since 16th century S. maxima reduced the fasting blood glucose levels by onwards. The metabolites of this cyanobacterium are of 40% and 75% respectively. The functional group analysis high therapeutic importance. of S. maxima revealed molecular vibrations specific for CH out of the plane stretch, CN- Stretch, C-O stretch, Ar C-C Objectives: The objective of the study was fractionation stretch, C=O and OH stretch. These are specific groups of and evaluation of antidiabetic potential of S. maxima and carbohydrates, proteins, aromatic compounds, lipids and identification of functional groups. polyphenolic compounds. The GCMS analysis of organic Methods: The anti-hyperglycemic potential of ethanolic extracts of S. maxima showed lipids of C5- C22 carbons. and aqueous extracts of S. maxima were evaluated in The fatty alcohols and isoprenoids were found to be 16.15 male Wistar rats injected with streptozotacin (45 mg/ and 30.32% in S. maxima while the fatty acids and fatty kg (i.p)). The molecular fingerprinting of S. maxima was acid conjugates constituted 54.98 and 20.86% respectively done using FTIR spectroscopy and the functional groups of total lipid content of S. maxima. were interpreted using IR PAL 2.0 and OMNIC 7.2a thermo Conclusions: From the study it is concluded that, the anti- library. The organic extract of S. maxima was derivatized hyperglycemic activity is associated with organic extract with diazomethane/BF3- methanol and analyzed through of S. maxima and the functional groups identified include GCMS. The mass spectrum was matched with the published fatty acids, fatty alcohols and isoprenoids. lipid databases.

EVALUATION AND MONITORING OF PREANALYTICAL ERRORS IN A HOSPITAL BASED LABORATORY

M. Kankra, S. Bhargava, A Manocha, S. Das, L.M. Srivastava Department of Biochemistry, Sir Ganga Ram Hospital, New Delhi, India

Background: Quality is the heart of management for all components of preanalytical phase from time to time in laboratories. For appropriate test results the entire testing an effort to improve the performance of the laboratory. process in lab is divided into three phases – preanalytical, Method: The study included an evaluation of preanalytical analytical and post analytical. The preanalyitical phase is quality indicator data collected from our lab over a period an important component of the total laboratory quality. of one year. A total of 2, 92,000 samples were received Errors in this phase are generally underestimated in routine of which 5,923 samples could not be processed as per practice. The aim of this study was to design and review

123 S94 Ind J Clin Biochem

our sample rejection policy. tion (10.3%) empty tube (9.8%), erroneous request (7.7%), error in patient identification (2.9%), and test order inap- Quality indicators for preanalytical phase were - inappro- propriateness (2.2%). priate sample (hemolytic, lipemic, clotted etc), insufficient sample, discordance between tests requested and test Conclusion: Sample management directly affects patient entered into computer system, erroneous request, error in care and outcome and also appropriately timed decision patient identification, test order inappropriateness, empty making of the clinician. Error tracking, scrutiny by evalu- tube and inappropriate transportation. ation and working constantly on preanalytical indicators are of utmost importance. With the aim of continuous Results: On evaluation of preanalytical indicators it was improvement the outcome of these indicators by correc- found that inappropriate sample was the most common tive measures over a period of time will definitely help error observed (36.7%) followed by discordance between to improve the quality of laboratory services and patient tests requested and test entered into computer system care. (18.3%), insufficient sample (12.1%), inappropriate transporta-

CA 199 REFERENCE RANGE IN DIABETES MELLITUS TYPE 2

Kahkasha, Bhawna Singh and Vinod Kumar Gupta Department of Biochemistry, Gobind Ballabh Pant Hospital, New Delhi, India

Introduction: CA 19-9 is a tumor-associated antigen that diabetic control subjects. Abdominal CT was performed in was originally defined by a monoclonal antibody (1116 NS order to eliminate abdominal malignancy in the diabetic 19-9) produced by a hybridoma prepared from murine and control groups. The serum levels of Ca19.9 and HbA1c spleen cells immunized with a human colorectal cancer were estimated by chemiluminometric assay performed cell line. CA 19-9 is elevated in pancreatic, upper gastroin- on cobas e-411(Roche) and BioRad D-10 HPLC analyser testinal tract, ovarian, hepatocellular and colorectal cancer respectively. The reference range for CA 19-9 in our lab and in certain non-malignant and inflammatory conditions being <39 U/Ml. of the lungs and hepatobiliary system. Results: HbA1C levels in diabetics compared from non Object of the study: To evaluate serum CA 19-9 levels diabetic control group were 10.2 ± 2.8 vs 5.2 ± 1.4 respec- in type 2 diabetic patients in comparison with age and tively. The mean CA 19-9 value was 49.8 ± 9.4 U/mL for gender-matched control subjects. diabetic patients whereas it was 10.2 ± 7.8 U/mL for the control group (P < 0.05). Materials and Methods: The case-control study was conducted in the department of Biochemistry, G B Pant Conclusion: The reference range of serum Ca19.9 is higher Hospital. Blood samples were collected from 40 known in diabetics which probably indicates that a higher cut-off cases of diabetes mellitus attending the OPD for routine value for CA 19-9 should be used for diabetics to eliminate follow up and 40 healthy, age and gender-matched non- doubts about pancreatic cancer.

PREVALENCE OF DIFFERENT TYPES OF HEMOGLOBINOPATHIES IN JHARKHAND

Bela Rose Ekka* Anupa Prasad** *Asstt Prof, Department of Biochemistry, RIMS, ** Tutor, Department of Biochemistry, RIMS.

Hemoglobinopathies are associated with an astonishing va- types of hemoglobinopathies in patients with hemtologi- riety of clinical and hematological manifestations. However, cal and clinical manifestations coming to paediatrics and most hemoglobinopathies are clinically and hematologically medecine department of RIMS. benign and detected only during screening programs. We performed HPLC in 80 such patients out of which A preliminary study was done for the presence of different 75% were having different types of hemoglobinopathies.

123 Ind J Clin Biochem S95

More than 50% patients had HbS in different quantities. double heterozygotes for HbS and β thalassemia. This group included Sickle cell disease, Sickle cell trait and

HEPATOPROTECTIVE ACTIVITY OF ETHANOLIC EXTRACT OF PHYLLANTHUS DEBILIS AND PLECTRANTHUS AMBOINICUS AGAINST ANTITUBERCULAR DRUGS INDUCED HEPATOTOXICITY IN WISTAR RATS

Don Mathew*, Shraddha hedge*, Ankita kaushal*, Krishnananda prabhu*, Smitha shenoy**, Mohan A**, Tara shanbhag**, Anand koppal**, Praveen Kumar**, Mohamed azar**, K. Praveen**, Ragavendra**, R. pirasantan** *Department of Biochemistry, KMC, Manipal University Manipal, India, **Department of Pharmacology, KMC, Manipal University Manipal, India

Introduction: Hepato-protectivity or anti–hepatotoxicity is of extracts of P.debilis [(200mg/kg, group III A),(400mg/ the ability of a substance to prevent liver damage. This kg, group IV A)] & P.amboinicus [(600mg/kg, group III B), study is aimed at evaluation of anti-oxidant property ef- (900mg/kg, group IV B)] & silymarin (50mg/kg, group V) fectiveness of using P.debilis and P.amboinicus extracts for daily for 30 days. treating hepatotoxicity induced by anti-tubercular drugs In Group VII (A+B), VIII (A+B) & IX, after 30 days dose of in Wistar rats. anti-tubercular drugs, rats received oral supplementation Materials & Methods: Plant extracts were prepared by of extracts of P.debilis [(200mg/kg, group VII A),(400mg/ thrice reflux condensation with absolute alcohol and dos- kg, group VIII A)] & P.amboinicus [(600mg/kg, group VII age was administered as suspension with 0.5% Carboxy B), (900mg/kg, group VIII B)] & silymarin (50mg/kg, group methyl cellulose. IX) daily from 31st -50th days.

A study group of 54 healthy Wistar rats (150-250g, either At the end of 30 days (Group I-V) & 50 days (Group VI- sex) were taken and divided into 9 groups of 6 each. XI), homogenised liver tissue was used for estimation of Group III, IV, VII, VIII were subdivided into A (3 rats) & Protein thiol (Ellman’s method). B (3 rats). Results & Discussion: The results will be discussed during Group I (30 days) received dosage of 0.5% Carboxy methyl presentation, as study is ongoing. cellulose (1ml/kg) daily. Group II (30 days) & VI (dose till Conclusions: Protein thiol levels can establish the potential day 30th, no drugs from day 31st -50th) received oral hepato-protective & anti-oxidant values of plant extracts dose of Isoniazid (31.5 mg/kg) + Rifampin (54 mg/kg) + of P.debilis and P.amboinicus for treating hepatotoxicity Pyrazinamide (189mg/kg) to induce anti tubercular drugs induced by anti-tubercular drugs. induced liver damage.

In Group III (A+B), IV (A+B) & V, after 30 days dose of anti-tubercular drugs, rats received oral supplementation

PREVALENCE OF ANTISPERM ANTIBODIES IN YOUNG URBAN INDIAN WOMEN PRESENTING WITH INFERTILITY

Rulie Buragohain, Prabhat Singh, R.K. Kapoor, SarikaArora, Shakti Agarwal Department of Biochemistry, ESI Hospital, Basaidarapur, New Delhi

Introduction: Infertility is a very common condition oc- for infertility is presence of antisperm antibodies in serum. curring more or less equally in both sexes. There are Antisperm antibodies are also found in cervical mucus, various causes of infertility and some cases of infertility seminal plasma and sera of men and women. The etiol- considered inexplicable have been found to have an im- ogy of sperm immunity in human females is unknown, but munologic basis. One of the immunologic factors proposed several possible mechanisms have been proposed, includ-

123 S96 Ind J Clin Biochem

ing cross-reactivity with microbial antigens and interferon Quantitative determination of Antibodies directed against gamma-mediated potentiation of the antisperm immune human spermatozoa were estimated in the serum by Elisa response in women whose male partners have sperm method. autoantibodies in their semen. Results: Total 57 infertile females were studied for presence The aim of this study was to find out the incidence of of antisperm antibodies. Out of which 35 were suffering antisperm antibodies in primary and secondary infertile from primary infertility & 22 were of secondary infertil- females coming to infertility clinic of ESI Hospital, Basaid- ity. Amongst the 35 primary infertility females 31.4% were arapur, NewDelhi. positive for ASA (11 out of 35) and in secondary infertility females the incidence of ASA was 22.7 % (5 out of 22). Objective: To evaluate the incidence of antisperm antibod- ies (ASA) and hormonal profile in infertile women. Conclusion: Antisperm antibodies exert heterogeneous effects on the ability of the spermatozoa to fertilize and Research Design and Methods: In the ongoing study, pa- may interfere with fertility. The study till now shows a tients registered for infertility treatment in the hospital, difference in the incidence of ASA in primary & second- irrespective of their etiologies, were screened for ASA. The ary infertility, the statistical significance of which will be study till now included 57 women suffering from infertil- determined after completion of the study. The detection of ity (either primary or secondary) for more than one year ASA can therefore provide a clue to the aetiologic factor duration. Detailed history of present and past illnesses as in such women which can aid in their management. well as medical and surgical treatment was taken.

BROAD SPECTRUM ROLE OF GINGER EXTRACT IN CASES OF PULMONARY TUBERCULOSIS PATIENTS

Dr. Subodh Kumar1, Dr. Mrs. Kiran Saxena2, Dr. Ravi Saxena3, Dr. K.R. Prasad4 1Department of Biochemistry, Major S D S Medical College, Fatehgarh, 2Department of Biochemistry, 3Department of Physiology, Chirayu Medical College & Hospital, Bhopal, 4Department of Biochemistry, Katihar Medical College & Hospital, Katihar (Bihar)

The study was designed to know the role of ginger ex- with respect to the initial stage (D - 0). tract in newly diagnosed sputum AFB positive Pulmonary The results of Anti-tuberculous treatment with ginger Tuberculosis patients having significant anemia. Total num- supplement (1st group) showed significant change in all ber of patients was 68 (sixty eight). The Patients were analyzed parameters i.e. CRP, Ferritin, Iron, EPO and He- randomly divided in two groups irrespective of age and moglobin (p < 0.001). Post-treatment (D - 30) levels of CRP sex. 1st group supplemented with ginger extract along and Ferritin & Erythropoietin were decreased whereas level with the anti-tuberculous treatment (n = 36) whereas of Serum Iron & hemoglobin were increased. Result of 2nd the 2nd group received only anti-tuberculous treatment, group patients who received only anti-tuberculous treat- which were serve as control. 250mg. of ginger extract ment showed similar pattern as in group 1st on day zero (equivalent to 1.5gm of pure ginger powder) was given but the (D – 30) results showed improvement which was orally twice daily after meals for 30 Days in capsular form statistically less significant as compared to that of group in both the groups. 1st. Beside this it was also observed potential decrease in In both the groups first batch of the blood samples were CRP level. At the same time there were significant rise in collected on zero day (D - 0) and 2nd batch of sample Hemoglobin level and weight gain by the patients in both was collected on thirtieth day (D - 30). Serum C Reactive the groups. But the result was more marked in the pa- Protein (CRP), Serum Ferritin, Serum Iron, Erythropoietin & tients with ginger supplement along with Anti-tuberculous Hemoglobin were estimated in all the samples. The results treatment group (1st), this was due to multifaceted, anti- obtained for these parameters were statistically analyzed tuberculous & hematanic action of ginger supplements. in both the patient groups. The results were compared Results of this study suggest that ginger can safely be

123 Ind J Clin Biochem S97

included in the standard anti tubercular treatment because pulmonary tuberculosis patients this was due to broad it maintains micronutrients level and provide immunity to spectrum role of ginger supplement.

INCREASED LEVEL OF NEOPTERIN ACCELERATE DYSFUNCTION OF NEUTROPHILS IN DEVELOPMENT OF ACUTE LIVER FAILURE

S. Prakash, Priyatma, H Sharma, Shalimar, Renu Yadav, S Bharti and SK Acharya Department of Gastroenterology and Human Nutrition, All India Institute of Medical Sciences, Ansari Nagar, New Delhi

Inflammation and increased oxidative stress (OS) are in- neutrophils was counted microscopically using the NBT volved in dysfunction of neutrophils and have synergistic method. Statistical analysis was done and p values <0.05 involvement with reactive oxygen species and releasing were deemed statistically significant. 76.5% abnormal neu- of neopterin may accelerate dysfunction of neutrophils trophils were counted in ALF while in healthy control it in development of acute liver failure. To evaluate serum was 16%. Significant decreased level of FRAP (p <0.000), neopterin levels and their correlations with OS markers in SOD (p <0.000), vitamin C (p <0.000) and MPO activity acute liver failure?. Abnormality of neutrophils function, OS (p <0.001) was found in ALF patients than healthy con- markers (free radical antioxidant potential (FRAP), Vitamin trol. An increased level of lipid peroxidation (p <0.000) C, Myeloperoxidase (MPO), Superoxide dismutase (SOD), and serum neopterin (p<0.001) were found in ALF than lipid peroxidation) and neopterin levels have evaluated in control subjects. SOD and FRAP levels were significantly 53 patients. The range of age was 22-65 years (M/F 4/1; (P<0.05) lower in sepsis while MDA and vitamin C was not 48.4 ± 18.8 years). The diagnosis of acute liver failure significant. As high neopterin production is associated with has done in the AB2 ward of Gastroenterology, AIIMS. increased production of reactive oxygen species and with Twenty age, sex matched (M/F 5/1; 33.23 ± 15.25 years) low serum concentrations of antioxidants like vitamin C, healthy controls have included in the study. In the etiol- SOD. Inflammation activation of macrophages in patients ogy of ALF clinical, biochemical and ultrasonographical with acute liver failure exhibiting elevated levels neopterin investigations was done in patients. Standard methodology may indicate neutrophil dysfunction. Thus, an assessment of have used for analysis of SOD, vitamin C, FRAP, MPO, neopterin status and functioning of neutrophils evaluation lipid peroxidation and serum neopterin. Abnormality of could be the diagnostic marker of ALF patients.

THYROID PROFILE IN GERIATRIC POPULATION

Dr. Abhijit Pratap, Dr. Mona A. Tilak, Dept of Biochemistry, Dr. D. Y. Patil Medical College, Pimpri, Pune

Introduction: The elderly population aged 70 and above, 60 – 65 years. which was only 8 million in 1961 rose to 21 million in 1991 Control group (group 2):- The control group consists of 30 and to 29 million in 2001. young ambulatory individuals of both sexes aged between Ageing leads to number of medical problems, which could 25 – 30 years. be attributed to various physiological changes occurring in TFT was measured by estimating the Total T3, Total T4 and all organs of the body. TSH in both the groups and comparing the results. Objectives: The aim of the study is to observe the thyroid TFT were also compared between males and females in parameter in geriatric age group. group 1 and group 2. Materials and Methods: Observation and Results: There is decrease in Total T3, Study group (group 1):- The study group consist of 30 Total T4 and increase in TSH (p < 0.05) in the geriatric elderly ambulatory patients of both sexes aged between population as compared to the young population.

123 S98 Ind J Clin Biochem

The Total T3 levels, is more in males as compared to Conclusion: This study concludes that there are differ- females in both geriatric and young population, however ences in the thyroid profile levels between the geriatric the Total T4 (p < 0.05) and TSH levels (p < 0.05) were population as compared to the younger population due more in females as compared to males. to various age related changes.

THERAPEUTIC DRUG MONITORING OF LAMOTRIGINE

Rohan Lokhande1, Prasad Naik1, A.J. Dherai1, Prajakta Kadam2, Parmee Dedhia1 Vrajesh Udani3, Roop Gursahani4, T.F. Ashavaid1 1Dept. of Laboratory Medicine, 2Research Laboratory, 3Dept of Pediatric Neurology, 4Dept of Neurology. P.D. Hinduja National Hospital & MRC, Veer Savarkar Marg, Mahim. Mumbai 400 016.

Introduction: Lamotrigine is an antiepileptic drug (AED) acid. Significant inter individual variability in drug level was used both as monotherapy and polytherapy. Its clearance observed. Therapeutic levels were obtained in 26 adults is significantly affected by coadministration of several drugs (7.8±3 ug/ml) and 34 children (7.7±4 ug/ml). A significantly and hence monitoring would help for dosage optimization higher level i.e. 9.4±3.6 ug/ml was obtained in adults on and therapeutic efficacy. polytherapy (n=15) as compared to that on monotherapy (n=21) i.e. 4.7±2.9 ug/ml. However in children lamotrigene Aim: To review lamotrigine levels obtained in the labora- level in polytherapy (n= 39)) and monotherapy (n=8) were tory and correlate the effect of dosage and polytherapy. overlapping i.e. 8.3±5.8 ug/ml and 8.0±4.4 ug/ml respec- Material and Methods: Pre-dose blood samples and de- tively. Irrespective of the drug regimen sub therapeutic tailed drug history were obtained from 36 adults and 46 level was observed in 9 adults and 7 children while toxic children on Lamotrigine therapy. Lamotrigine estimation level were observed in 1 adult and 5 children. Dosage was done by HPLC. The relationship between concentra- modifications in these patients resulted in optimal level tion and dosage was assessed. Follow up drug levels after on follow up analysis. dosage modification was performed in 19 patients. Conclusion: Lamotrigine monitoring helps in assessing Result: Monotherapy was administered in 17% of children and maintaining the dosage in individuals on mono and and 43% of adults. Polytherapy mainly included valproic polytherapy to obtain optimal epilepsy control.

DYSLIPIDAEMIA AND OXIDATIVE STRESS IN APPARENTLY HEALTHY LATE ADOLESCENT AND YOUNG ADULT PASSIVE SMOKERS

Viyatprajna Acharya*, S Pradeep, S Lakshmiprabha

Introduction: Passive smoking is considered to be equally or Objective: A case-control study was done to learn about more hazardous than active smoking due to the exposure the oxidative stress and extent of cardiovascular risk in to the side-stream smoke which contains certain toxins the passive smokers. in more amount than the main stream smoke. Passive Methodology: 50 apparently healthy passive smokers and smoking is responsible for 1% deaths around the world and 50 non-smokers in the age group 16-25 years were chosen about 10% of such passive smokers reside in India. and their routine investigations, lipid profile, serum uric acid Due to direct and indirect stimulation of sympathetic and oxidative stress marker MDA (Malondialdehyde) were system it leads to dyslipidaemia or affects heart directly. assayed to see the correlation between passive smoking, The different chemicals present in side-stream of smoke oxidative stress and cardiovascular risk factors. are either oxidants themselves or may lead to generation Results: In passive smokers total cholesterol and MDA were of free radicals that ultimately gives rise to cardiovascular statistically significantly high (p< 0.001). MDA was found disorders.

123 Ind J Clin Biochem S99

to be correlated to TC (r=0.33; p< 0.01), LDL-C (r=0.35; p< 0.001). Passive smoking exposure was significantly associ- 0.01), Non- HDL-C (r= 0.3; p<0.01). Also MDA was found ated with MDA (OR=17.93, p<0.0001; CI=95%). to be positively correlated to SBP, DBP and respiratory Conclusion: Passive smokers have a potential risk to de- rate that were statistically significant. Passive smoking was velop oxidative stress and dyslipidaemia which may lead found to be strongly associated with hypercholesterolae- to cardiovascular disorders in the long run. mia (OR=17.2; p< 0.001) and oxidative stress (OR=38.5; p<

ELEVATED SERUM TOTAL TESTOSTERONE LEVELS AND THEIR RELATIONSHIP WITH HYPERTENSION IN PREECLAMPSIA

Vinayak W. Patil *, Shahid A. Mujawar **, Rekha G. Daver*** *Professor and Head, ** Assistant Professor, Department of Biochemistry, ***Professor and Head, Department of obstetrics & Gynecology, Grant Government Medical College & Sir J.J. Group of Government Hospitals, Byculla, Mumbai

Background: Most current hypotheses regarding the Results: The statistical analysis of study group of preec- pathophysiologic mechanisms of preeclampsia point to lampsia compared with normotensive control group, trophoblastic dysfunction. Therefore, we attempted to showed significant increase in serum TT concentrations in evaluate the relationship between serum total testosterone patients with preeclampsia. (TT) and hypertension in patients with preeclampsia. Conclusion: The present study found a significant associa- Material and method: Fifty preeclamptic patients from tion between serum TT and mean blood pressure as a risk gynecology ward were studied for estimation of serum factor for cardiovascular disease (CVD) and trophoblastic TT over a period of October 2007 to June 2010. Serum dysfunction in preeclampsia. TT determined by means of fully automated enzyme Keywards: Total Testosterone, Mean Blood Pressure, amplified chemiluminescent immunoassay based Immulite Preeclampsia. 1000 analyzer. Measurement of serum TT by using com- mercial kits from Siemens Medical Solutions Diagnostics, Los Angeles, CA, USA.

THE INFLUENCE OF GENERAL MATERNAL WEIGHT ON SERUM ALPHAFETOPROTEIN AND FREE BETAHUMAN CHORIONIC GONADOTROPIN LEVELS DURING THE SECOND TRIMESTER OF PREGNANCY

Diganta Dey, Abhijit Banerjee, Parbati Banerjee, Arnab Paul and Ratnamala Ray Dept. of Biochemistry, Ashok Laboratory Clinical Testing Centre Pvt. Ltd., Kolkata

Objective: Maternal serum alpha-fetoprotein (AFP) and tion, during January’2010 to July’2012, using enzyme linked free beta-human chorionic gonadotropin (beta-hCG) levels immunosorbent assay (ELISA; AFP-hCG-uE3 AccuBind VAST are useful biomarkers for antenatal screening of Down ELISA Kit, USA), followed by the PregSafe prenatal risk syndrome, Trisomy 18 and open neural tube defects. The assessment software. objective of this study was to investigate the relationship Results: The mean maternal weight was 59.75 ± 18.02 kg between maternal weight and serum AFP and free beta- in studied pregnant women during the second trimester. hCG concentrations. There was a markedly inverse relationship between mater- Methodology: The study included 70 normal singleton nal weight and serum AFP and free beta-hCG levels. The pregnancies (mean age: 31.45 years) without any congenital median AFP and free beta-hCG level for women weighing anomalies, were screened using AFP, free beta-hCG and ≤ 50 kg were respectively 90.4% and 57.4% higher than unconjugated estriol between 16 and 20 weeks of gesta- the median level for women weighing 70 kg or more. The

123 S100 Ind J Clin Biochem

overall variation in AFP (p = 0.17) and free beta-hCG (p = Conclusion: Maternal weight may affect the AFP and free 0.69) levels were significant between these two weight beta-hCG concentrations during the second trimester. Heavi- groups. The reference interval (2.5 to 97.5 percentile) for er pregnant women have lower median values, perhaps AFP and free beta-hCG were obtained 5.56 – 136.37 ng/ as a result of a diluting effect of larger blood volume. A mL and 4.41 – 283.55 IU/mL respectively. revision of the reference interval for AFP and free beta-hCG considering the body weight might improve in assessing risk of poor outcome, especially in heavier women.

STUDY OF ANTIOXIDANT LEVEL AND SERUM CREATININE IN HYPERTENSIVE DISORDER OF PREGNANCY

Sonali Singh, Dr. Arun Shrivastva, Dr. Tripti saxena, Dr. Shabana Sultan. Dr. Bhawana Bhimte Department of Medical Biochemistry, Department of Gaynecology, Gandhi Medical College, Bhopal

Introduction: Hypertensive disorders of pregnancy is a disorders of pregnancy (pre-eclampsia & eclampsia. The common complication of late pregnancy is a major cause present study includes 50 patients of preeclampsia and of renal damage during pregnancy. It includes: Pre-ec- eclampsia. 50 healthy controls. Blood sample were tak- lampsia and Eclampsia. Pre eclampsia general prevalence ing and following investigation were done estimation of is between 2-3% of pregnancy. It is a common cause of serum creatinine by Jaffo reaction without deproteinisa- both maternal and perinatal morbidity & mortality in both tion, kinetic method. And Estimation of serum uric acid developed and developing countries. Eclampsia, an acute by Uricase/Trinder method. and life-threatening complication of pregnancy. It includes Results: This study showed the significant increased in seizures and coma that happen during pregnancy but are serum uric acid and, significant increased in creatinine not due to preexisting or organic brain disorders. cases as compare to healthy controls. Objective: To study of serum creatinine and serum uric Conclusion: Increased Serum uric acid and serum creati- acid level in hypertensive disorder of pregnancy. nine level in associated with in hypertensive disorders of Methods: The aim of this study was the assessment of pregnancy (pre-eclampsia and eclampsia). serum creatinine and serum uric acid level in hypertensive

URINARY CALCIUM/CREATININE RATIO IN THE PREDICTION OF PREECLAMPSIA

Dr. Mittal Shilpa, Dr. Shaikh M.K.S., Dr. Thakur R., Dr. Verma M., Dr. Meena M. SAIMS Medical College & P. G. Institute, Indore

Objective: The objective of the study is to evaluate pre- method & Creatinine by Jaffe’s method by HITACHI 902 dictive values of Urinary Calcium/creatinine ratio in Pre- Autoanalyser. eclampsia and to introduce spot urine test in ANC profile. Result: In this clinical study the mean Urinary Calcium/ Pre-eclampsia is an idiopathic multisystem disorder causes creatinine ratio is lower in Pre-eclampsia (i.e.; less than or maternal and fetal morbidity and mortality. equal to 0.04) compared with normotensive group. The Materials and Methods: The present study was conducted incidence of Pre-eclampsia was 13.63% out of which 11.36% in the Departments of Bio-chemistry and Obstetrics & Gy- is mild, 2.27% is moderate Pre-clampsia. necology, SAIMS Medical College & P. G. Institute, Indore. Conclusion: This study evaluates that a single Urinary Cal- The study includes total 44 asymptomatic pregnant women cium/creatinine ratio in spot urine sample can be added as aged between 21-35 years with 20-28 weeks of gesta- an effective and routine marker in ANC profile for predict- tion. Urinary Calcium was estimated by O-Cresolphthalein ing Pre-eclampsia in a high risk population.

123 Ind J Clin Biochem S101

PLASMA PROTEIN CARBONYL AND RBC REDUCED GLUTATHIONE LEVEL IN BETA THALASSEMIA MAJOR PATIENTS ATTENDING A TERTIARY CARE HOSPITAL IN EASTERN INDIA

Tanima Mandal Department Of Biochemistry, Calcutta National Medical College, Kolkata

Thalassemia refers to a heterogeneous group of inherited oxidant and anti oxidant status in these subjects. Studies disease characterized by defect in synthesis of alpha or for beta thalassemia have shown increased plasma protein beta globin chains of haemoglobin. There is collaboration carbonyl with reduced RBC Glutathione between oxidative stress and anaemia in thalassemia. Re- 100 diagnosed cases of beta thalassemia major based on active oxidative species (ROSs) are generated in increased clinical features, HPLC and haematology based laboratory amount in thalassemic erythrocytes and oxidative insult in findings and receiving transfusion management at thalas- thalassemia leads to haemolysis. Carbonyl stress represents semia clinic of the above mentioned institute has been toxic effect of ROSs and various mono di carboxyl and randomly selected. 100 age and sex matched controls alpha di carboxyl molecules on proteins. The evaluation attending the immunization clinic has been also included of carbonyl group content in blood protein is considered in the study. Plasma protein carbonyl estimation by Di as a useful biomarker of oxidation induced by Glutathi- nitro Phenyl Hydrazine (DNPH). RBC reduced glutathione one is a sulphydryl group containing tri peptide, acting estimation by 5,5’ di thiobis 2 di thiobis nitro benzoic acid as an antioxidant, that prevents cellular damage caused (DTNB). Obtain data shall be co-related each other and by ROSs. In this background a study has been designed draw a suitable conclusion. assess plasma protein carbonyl and reduced RBC glutathi- one label in beta thalassemia major cases to assess the

APA I POLYMORPHISM AND HBF LEVELS IN BETA THALASSEMIA PATIENTS OF THE GWALIOR CHAMBAL REGION OF CENTRAL INDIA

Rakesh Kumar1, Dharmesh C Sharma2, Purnima Kishor1 1School of studies in Biochemistry, Jiwaji University, Gwalior, 2Blood Bank, Jayarogya Hospital, GR Medical College, Gwalior

The treatment of beta thalassemia, especially in developing promoter of regularly transfused β-thal patients of the countries, is far from satisfactory. Because of the milder Gwalior Chambal region of Central India. The subjects phenotypes often associated with relatively high levels were tested for the presence Apa1 polymorphism by PCR, of fetal hemoglobin in patients with different forms of β and its association with HbF levels, determined by HPLC, thalassemia there is major interest in the factors that regu- as well as the association of the ApaI site with beta late the switch from fetal to adult hemoglobin and how globin gene mutation. 11% patients presented with Apa1 this may be modified for therapeutic purposes. Recently polymorphism of which only one exhibited an increase in there has been considerable progress in defining the loci HbF. HbF levels of the other Apa1 positive patients were or genomic regions that may be involved in the regulation in the normal range. The ApaI site was associated with of the switch from fetal to adult hemoglobin. all the six mutations studied. Thus, the present study reveals that in the subjects of this region, no correlation ApaI polymorphism (−202 (G›T)), in A γ-globin gene exists between ApaI polymorphism and HbF levels and promoter is known to affect gamma globin gene ex- that the site is not associated with any particular beta pression and ameliorate disease symptoms. The present globin gene mutation. study involves an analysis of 120 alleles of Aγ-globin gene

123 S102 Ind J Clin Biochem

SIX SIGMA IN THE HEALTH CARE SECTOR : QUALITY IMPROVEMENT IN THE CLINICAL LABORATORY OF A TERTIARY CARE HOSPITAL

Dr. Leena Dash Consultant Biochemist and Laboratory Head (Certified Black Belt in Six Sigma), SRL at Fortis Hospitals, Bannerghata Road, Bangalore

Introduction: To address concerns of delayed Turnaround sigma methodology. Time for routine Biochemistry and Hematology tests in Key steps of the Lab processes and sub processes were the clinical laboratory, a quality improvement project, mapped with MS Visio. Pareto analysis helped to identify using Six Sigma Methodology was used to correct issues the steps which were responsible for maximum delays. leading to the delay. Statistical calculations were done with Minitab software. Objective: To reduce TAT for routine testing in the Labora- Results: The initial average TAT was 137.4+/-15 with almost tory and to put a process for effectively implementing the 90 % of tests crossing the target TAT of 90 minutes. same for all hospital patients. This would increase patient From this level, the operating average TAT was brought and clinicians satisfaction as it would reduce patient wait- down to 71+/- 15 minutes with only 20 % of tests with a ing time, faster consultations and improved discharge time longer TAT. for inpatients. The required TAT for effective services was defined as 90 minutes. Conclusions: The study was carried out for a year and is ongoing, with weekly data monitoring. It highlighted the Methods: The project was carried out in a 250 bedded, effective use of Six sigma analyses in the healthcare sec- multispecialty hospital in Bangalore; Karnataka during a tor for improving care processes, eliminating waste and period of June 2011 to June 2012. A team was put together enhancing customer satisfaction. for handling each phase of the DMAIC steps used in six

COMPARISON OF BLOOD GLUCOSE ESTIMATION ON THREE DIFFERENT INSTRUMENTS

Harjeet Kaur, Jasbinder Kaur, Shivani Jaswal, Seema, Sumita Sharma Department of Biochemistry, Government Medical College & Hospital, Chandigarh

Introduction: Although various instruments have been 3 instruments (fully automated analyzer ROCHE MODULAR developed for estimation of blood glucose, not much P 800, metabolite analyzer RADIOMETER ABL 800 FLEX is explored about their accuracy and comparability. The and semi-autoanalyzer ERBA CHEM 5 PLUS) concurrently. accurate measurement of glucose is extremely important According to the blood glucose levels, the samples were in the diagnosis of diabetes and pre-diabetes, where the divided into 3 groups (<80mg/dl, 80-110mg/dl and >110mg/ laboratory values are crucial. Tight control of plasma dl) of 25 samples each. The results of blood glucose values glucose is now a mandate for diabetes treatment. This, were compared with those obtained from the commonly combined with the lower diagnostic criteria for diabetes used routine instrument i.e. semi-autoanalyzer. Analyses of (> 126 mg/dl FBG) and the new category of pre-diabetes data were performed with the student’s t test. starting at an FBG of 100 mg/dl, our ability to rely on the Results: Mean glucose concentrations on fully automated accuracy of glucose measurement is clearly important. We analyzer, metabolite analyzer and semi-autoanalyzer therefore compared the best new instruments with the were 82.2±8.67 mg/dl, 75.5±7.4mg/dl and 73.0±4.9 mg/ former instrument being used in most of the laboratories dl respectively for group with glucose conc. of <80mg/ that is the semi-auto analyzer. dl, 102.4±15.7mg/dl,100.4±12.33mg/dl and 94.4±9.0 mg/dl Methods: Fasting blood samples of 75 subjects were col- respectively for group with glucose conc. of 80-110 mg/dl lected into sodium-fluoride tubes, and glucose levels were and 182.0±62.7mg/dl, 175.4±62.9mg/dl and 166.0±59.9 mg/dl measured in plasma. The measurements were obtained on respectively for group with glucose conc. of >110 mg/dl.

123 Ind J Clin Biochem S103

On comparing the results of automated analyzer with Conclusion: From this study, it is concluded that differ- the semi autoanalyzer, it was found that the result on ences in glucose levels measured by using various types automated analyzer were 10.2% higher than those on semi of instruments are not negligible and can not be ignored. autoanalyzer. Similarly the results were found to be 5.1% So we have to identify the most precise instrument for higher on radiometer as compared to semi autoanalyzer. estimation of blood glucose. This difference was found to be statistically significant (p<0.001)

PODOCYTURIA A NEW MARKER FOR EARLY DIABETIC NEPHROPATHY

Dr Brijesh Mukherjee, Dr Sibasish Sahoo Department of Biochemistry, Kalinga Institute of Medical Sciences, Bhubaneswar

Constant efforts are ongoing to identify reliable and re- from that in Europeans in several aspects: the onset is at producible noninvasive biomarkers for acute and chronic a younger age, obesity is less common, and genetic factors kidney injury targeted toward identifying kidney injury not appear to be more common. Some studies conducted in only in its early stages, but also in classifying kidney dis- migrant Asian Indians in the U.K. and Europe have reported ease according to severity, predict disease outcomes, and increased prevalence of diabetic nephropathy compared monitor response to therapeutic interventions. with white Caucasians. The few studies published on the prevalence of diabetic nephropathy in India have all been The number of podocytes in urine or podocyturia increases clinic based. Indeed, the Diabetes Atlas 2006 does not list with active kidney disease even before proteinuria appears a single population-based study on diabetic nephropathy and seems to improve with treatment. Also, podocyturia from South Asia. seems to be confined to active disease, in contrast to pro- teinuria, which is present during both active and chronic Diabetes is becoming the major single cause of end stage phases of glomerular damage. It will be particularly inter- renal disease in the world. Precise evaluation of renal func- esting to explore podocyturia, as a marker of subclinical tion in early stages of diabetic kidney disease may indicate early renal damage, which may be a detectable way before patients susceptible to progression to the end-stage renal the occurrence of overt proteinuria and development of disease. Intensification of the treatment in those patients full blown glomerular disease. might slow the progression of the disease. Implementation of behaviour modification and pharmacological therapy Diabetic nephropathy is the leading cause of end-stage targeted especially at hyperglycemia and hypertension renal disease (ESRD) worldwide, and it is estimated that declines urinary albumin excretion rate and decreases the ~20% of type 2 diabetic patients reach ESRD during their progression to end-stage renal disease. Intensive treatment lifetime. Kidney disease in diabetic patients is clinically is associated with increased life expectancy and is more characterized by increasing rates of urinary albumin excre- cost-effective than conventional treatment in patients with tion, starting from normoalbuminuria, which progresses diabetic kidney disease Progressive renal function decline to microalbuminuria, macroalbuminuria, and eventually to in diabetes is an early event that occurs in a proportion ESRD. Microalbuminuria is the earliest clinically detectable of patients without increased albumin excretion rate. stage of diabetic kidney disease at which appropriate interventions can retard or even reverse, the progress of Estimation of the glomerular filtration rate (GFR) is the the disease. most widely used test of renal function and reflects the kidney’s ability to clear a particular substance from plas- According to the most recent estimates published in the ma. GFR is defined as the quantity of glomerular filtrate Diabetes Atlas 2006, India has the largest number of dia- formed per unit time in all nephrons of both kidneys. The betic patients in the world, estimated to be ~40.9 million most precise and accurate methods for estimating GFR in the year 2007 and expected to increase to ~69.9 million are based upon determinations of plasma clearance of by the year 2025. Type 2 diabetes in Asian Indians differs substances like 51Cr-EDTA, iothalamate or iohexol. These

123 S104 Ind J Clin Biochem

so called “gold standard” methods require injection of an to undergo regenerative proliferation to compensate for a exogenous radioactive or contrast agent, and are complex, loss of podocytes or an increase in GBM surface area. It laborious, expensive and impractical in the clinical setting has been speculated that podocyte cell cycle quiescence is and for larger research studies. Therefore, the measure- a prerequisite for a functional glomerulus as for podocytes ment of endogenous blood substances to estimate GFR reentry into the cell cycle a deconstruction of the highly is common practice. For several decades clinicians have structured cytoskeletal organization would be required with relied on measurements of serum creatinine as a rapid loss of glomerular permselectivity. In human DN there is first-line test to determine GFR. This test is convenient a decrease in the number of podocytes per glomerulus. and cheap, but results are affected by age, sex, muscle Furthermore, loss of podocytes correlates closely with the mass, diet, race and tubular creatinine secretion, particularly degree of progression with fewer podocytes per glom- when GFR is reduced. Thus there is a need for ongoing erulus in the rapidly progressing group compared with research for identification of new markers for early diabetic slow progressors. It is unclear whether in DN there is nephropathy. an absolute reduction in podocytes number or a relative reduction due to increased GBM surface area. In the first Traditionally, the appearance of microalbuminuria has been case, podocytes structural alterations and damage would used to detect the onset of diabetic nephropathy (DN), be the primary event resulting in podocytes apoptosis and and its appearance prompts aggressive treatment. podocyturia. In the second case, both glomerular hyper- However, podocyte injury starts to occur in previously as- trophy and enhanced glomerular volume expansion would sumed unaffected populations of patients before microal- cause an increase in GBM surface area with consequent buminuria appears, and therefore podocyturia is an earlier reduction in podocytes density. FP broadening would be a marker of DN than microalbuminuria, researchers stated compensatory response of the remaining podocytes in the here on April 15 at the National Kidney Foundation (NKF) attempt to cover areas of bare GBM. Broadening, however, 2010 Spring Clinical Meetings. reduces FP adhesion to the GBM and may eventually result in podocyte detachment and loss. Podocytes are highly differentiated cells with a complex cel- lular morphology. Podocyte cell body bulges into the urinary Podocytes are normally absent or seen in small numbers space and gives rise to long primary processes that extend in urine of normal individuals or those with inactive kidney toward the capillaries to which they affix by numerous disease. Although not visible utilizing a microscope, it is foot processes (FPs). The FPs of neighbouring podocytes possible to visualize these podocytes in urine with immu- interdigitate, leaving between them filtration slits bridged nofluoresence staining and after incubation with antihu- by an extracellular structure, known as the slit diaphragm man podocalyxin monoclonal antibody PHM-5 (Australian (SD). Our knowledge of the molecular structure of the SD Monoclonal Development, Artarmon, New South Wales, has remarkably improved in the past few years. Several Australia). The number of podocytes in urine or podo- molecules, including nephrin, CD2AP, FAT, ZO-1, P-cadherin, cyturia increases with active kidney disease even before Podocin, and Neph 1-3 have been shown to be associ- proteinuria appears and seems to improve with treatment. ated with the SD complex, and some of these molecules Also, podocyturia seems to be confined to active disease, are now known to be critical for its integrity. Podocyte in contrast to proteinuria, which is present during both structural abnormalities and loss have been described in active and chronic phases of glomerular damage. It will be DN. Furthermore, podocytes form an integral part of the particularly interesting to explore podocyturia, as a marker glomerular filtration barrier and participate in GBM forma- of subclinical early renal damage, which may be a detect- tion, and thus they may be implicated in GBM thickening able way before the occurrence of overt proteinuria and and abnormal glomerular permeability to proteins. Primary development of full blown glomerular disease. podocytes undergo dedifferentiation and apoptosis in cul- NOTE: Fresh urine is collected and urinary cell pellets were ture ; however cell lines of differentiated podocytes have derived via centrifugation. Immunofluorescence is used to recently become available, allowing the in vitro study of mark the presence of podocytes. The urinary podocytes are podocyte response to the insults relevant to DN Podocyte identified by co localisation of podocyte-specific markers. Apoptosis-Loss In the adult kidney podocytes are unable The podocytes-to-creatinine ratios can also be calculated in

123 Ind J Clin Biochem S105

order to semi quantify the cases of podocyturia sediments of patients with various forms of glomerulone- phritis by immunoflorescence and reported that the number A glomerular epithelial cell antigen, podocalyxin, has been of podocytes in the urine was a direct indication of the found in the urine of patients with glomerular disease. Hara degree of glomerular epithelial cell injury et al. detected podocalyxin-marked podocytes in urinary

INSULIN RESISTANCE ACCOMPANIES TSH RESISTANCE IN OBESE

Kiran Chugh1, Sandeep Goyal2, Vijay Shankar1, S.N. Chugh2, Yuthika Agrawal1 1Department of Biochemistry, 2Department of Medicine, Pt. B. D. Sharma University of Health Sciences, Rohtak, Haryana.

Background and Objectives: Obesity is associated with vari- T3, T4 were done using respective radioimmunoassay kits. ous endocrinal and non endocrinal disorders. One major Homeostasis model for insulin resistance (HOMA IR) was endocrine disorder associated with obesity is insulin resis- calculated and cases were divided into insulin resistant tance, due to defect in post receptor signal transduction [subgroup IA] and insulin non-resistant [subgroup IB] in target tissue. Also obesity causes increase in thyroid based on standardised value (mean+2 S.D.) of HOMA IR stimulating hormone (TSH) with no effect or increase in of controls [group II] which was taken as reference value triiodothyronin (T3), and no effect on thyroxin (T4). On to define the insulin resistance. the other hand, subclinical hypothyroidism causes obesity Results: TSH was significantly increased in insulin resistant due to slow metabolism. Thus thyroid function tests and than in non resistant, whereas T3 and T4 levels were insulin resistance were evaluated to know whether TSH comparable. increase is a cause or consequence of obesity. Conclusion: Raised TSH levels along with comparable and Methods: 50 obese cases (body mass index > 30 kg/m2) normal T3 and T4 in obese persons with insulin resistance [group I] were taken against 25 non obese controls (body could be due to association of thyroid receptor resistance mass index < 25kg/m2) [group II]. Blood sugar, insulin, T3, to TSH similar to insulin resistance. T4, TSH levels were measured. Blood sugar was estimated by glucose oxidase-peroxidase method while insulin, TSH,

A CORRELATION OF THE CONCENTRATION OF FLUORIDE IN BLOOD AND INCREASED LIPID PEROXIDATION AND ASSOCIATED COGNITIVE DECLINE IN FLUOROTIC PATIENTS OF RAJASTHAN

Vivek Pratap Singh1, Vikas Gaur2, Mukesh Tiwari3, Dushyant S. Chauhan1, Sudhanshu Mishra1, Sandeep Kumar1, Sandeep Tripathi1, Anurag Tomar1,4 National Referral Centre for Fluoride Poisoning in India, (NRCFPI). 1Department of Biotechnology, Nims Institute of Engineering & Technology; Departments of 2Psychiatry, 3Orthopedics and 4Pediatrics, Nims Medical College & Hospital. Nims University, Shobha Nagar Jaipur Rajasthan, Jaipur

Background: The data has been suggested that Fluorosis Method: In the present study, 57 adults (age, 20 to 40 in Rajasthan has gradually increased due to the presence years) fluorotic patients were selected from the high fluo- of higher amount of fluoride in drinking water (>1.5ppm). ride region of the eastern regions of the Jaipur, where It may be serious problems in health of the adults and fluoride content in water is 5.5 ± 1.2 ppm. Moreover, age children. In the state of Rajasthan, almost all districts have matched controls were selected from the Jaipur district high Fluoride (up to 24 ppm) in their drinking / ground where fluoride content in water was less than 1.5 ppm. water sources and about 11 million of the populations are 5.0 ml of blood sample were taken for estimation of fluo- at risk. The aim of the present study was to establish ride and lipid peroxide levels. The GHQ60 questionnaire a correlation between behavioural changes and fluoride was used to assess behavioural changes in patients and content in the blood. healthy subjects.

123 S106 Ind J Clin Biochem

Results: Significantly increased rate of lipid peroxidation increased production of malenodyaldehyde (MDA) in blood. and depleted in antioxidant levels superoxide dismuatase, These alterations may induce pathophysiological activities catalyse, glutathione peroxidase and glutathione (SOD, CAT, due to lack of antioxidants. Moreover, perturbed behav- GPX and GSH) was observed in patients. The concentration ioural activity score directly proportion to the concentra- of fluoride was significantly (p<0.0001) elevated and it’s tion of fluoride and rate of lipid peroxidation. However, correlated with the altered behavioural score (GSHQ60). further in depth studies is required for the understanding of pathophysiology of fluorosis. Conclusion: On the basis of the results it may conclude that fluoride exposure promote oxidative stress through

STUDY 0F SERUM INTERLEUKIN6 IL6 AND TUMOR NECROSIS FACTOR ALPHA TNF Α IN POST MENOPAUSAL OSTEOARTHRITIC FEMALE PATIENTS

Pradeep Sharma Department of Biochemistry, U.P. Rural Institute of Medical Science & Research Saifai Etawah

Abstract: Osteoarthritis is a very common chronic disease age group. The IL-6 and TNF-α were measured in the that affects all joint tissues, causing progressive irrevers- serum by enzyme linked immunosorbent assay technique. ible damage and finally, the failure of the joint as an TNF-α and IL-6 levels were high as compared to control organ. The present study is to determine the levels of female subjects (P<0.001). TNF-α and IL-6 responsible interleukin-6 and tumor necrosis factor alpha are associ- for the degradation of cartilage in osteoarthritis in post ated with osteoarthritis in post menopausal females. The menopausal females. present study is being carried out in 150 female subjects Keyword: TNF-α, IL-6 and osteoarthritis of age group of 45-60 years suffering from osteoarthritis and 50 normal healthy control female subjects of same

LIPID PROFILE AND BLOOD SUGAR LEVEL IN PSORIASIS PATIENTS

Monika Mandrai*, Dr. B.K. Agarwal* Dr. Tripti saxena*, Dr. Bhawna Bhimte*, Dr. Anna Alex**, Harish Makwane Department of Medical Biochemistry, Gandhi medical College, Bhopal

Introduction: Psoriasis is a chronic disorder characterized The present study includes 50 patients of psoriasis and 50 by erythematous scaly patches that affect the scalp, trunk, healthy controls. Blood sample were taking and following extensor surfaces of the limbs and the genital area. Psoria- investigation were done: - estimation of serum lipid profile sis is highly prevalent in the general population, mainly as by GPO/POD, CHOD-PAP method. Estimation of blood sugar a result of its chronic nature and the absence of a cure. level by GOD/POD method. Psoriasis affects about 2–4% of the general population. In Results: This study showed the significant increased to- Israel, the prevalence of psoriasis is estimated at 2% (Israel tal cholesterol and LDL cholesterol, significant increased Society for Dermatology and Venereology, Personal Com- blood sugar level in psoriasis cases as compare to healthy munication). Psoriasis is associated with decreased quality controls. of life and high rates of depression. Patients with mild to moderate psoriasis are usually given topical treatments. Conclusion: In conclusion, increased Serum Cholesterol Systemic therapy or phototherapy is reserved for patients and LDL-cholesterol level in associated with in psoriasis. with moderate to severe psoriasis. Increased significant blood sugar level in psoriasis pa- tients. Methods: The aim of this study was the assessment of serum lipid profile and blood sugar level in psoriasis cases.

123 Ind J Clin Biochem S107

MEASURING THE EFFECT OF DIFFERENT PARAMETERS ON THE CATALYTIC ACTIVITY OF IMMOBILIZED ΒGALATOSIDASE FROM YEAST

Sandhya Hora1, Manoj Gupta2, Mamta Gupta3 1Research Assistant, Central Research Laboratory, Department of Biochemistry, Shri Ram Murti Samarak-Institute of Medical Sciences, Bareilly, Uttar Pradesh. 2,3PG Students, Department of Biochemistry, Shri Ram Murti Samarak-Institute of Medical Sciences, Bareilly, Uttar Pradesh.

Background: Enzyme catalysis is intermediate between Results: The effect of temperature, pH, and amino acids homogeneous and heterogeneous catalysis. In general, on normal β-galatosidase and on immobilized β-galatosidase it is similar to inorganic heterogeneous catalysis and were analyzed. Immobilized enzyme showed the decreasing sometimes it is called micro heterogeneous catalysis. Ef- conversion level of lactose. Polyacrylamide beads showed ficient enzyme catalysis depends on high specific nature decreasing conversion level of lactose at different concen- of enzyme, pH, temperature, optimum temperature, col- tration of lactose and same concentration of beads. Effect loidal nature, activator or co-enzyme, inhibitors or poisons. of temperature on β-galactosidase was maximum at 400C Immobilized β-galatosidase show high activity towards for growth. pH profile (6.2 to 7.4) of β-galactosidase was both pure lactose and lactose in skim milk, and a better studied at 280C & 370C. Highest lactose conversion (4.44%) thermal stability also. was noted at pH-6.8.

Methods: β-galatosidase was extracted by Kluyveromyces Conclusion: It was found that the β-galactosidase from lactis and enzyme assay done by GOD/POD method. Partial K.lactis showed an optimum activity at temperature 400C, purification of enzyme was done by precipitation with am- pH 6.8. Permeabilization of the cell wall with chloroform monium sulphate, DEAE-cellulose column chromatography increased the enzyme activity. The immobilized non vi- followed by SDS-PAGE. Immobilization of K.lactis cells was able permeabilized cells give better result then viable done by Entrapment method in which we agarose beads, immobilized cells. plyacrylamide beads and alginate beads were used to analyze the catalytic activity of β-galatosidase on differ- ent parameters.

STUDY OF SERUM E SELECTIN LEVELS IN CARDIOVASCULAR DISORDERS IN UDUPI DISTRICT

Salini Shereen Koshy, Sumanth H.D., Namita Rao, Varashree B.S., Pragna Rao Department of Biochemistry, Kasturba Medical College, Manipal University, Manipal, Karnataka

Objective: To correlate the known pro-atherogenic risk Result: E-selectin showed a negative correlation with hs- factors like low HDL levels with E-selectin levels in angio- Troponin- t in Group 2. There was a negative correlation genesis and cardiovascular disease, and to see if E-selectin between hs- Troponin- t, E-selectin and HDL in group 3. levels in blood can be used along with other parameters Conclusion: There does not appear to be any role for E- to screen for risk of CVDs. selectin as a marker in the studied population according Methods: Serum samples of subjects between the ages to the present study. The fact that E- selectin has genetic of 50-60 with CVD were chosen randomly on the basis polymorphism may explain the lack of significant difference of hs- Troponin- t levels and were divided into 3 groups in E-selectin levels. of 15 each. The Abnova human sE- selectin immunoassay- a four and a half hour solid phase ELISA was used to measure sE- selectin in serum samples. CK, CKMB- mass, hs- Troponin- t and HDL were analysed using auto analy- ser- Cobas 6000.

123 S108 Ind J Clin Biochem

HOMOCYSTEINE AND VITAMINS

Dr. Madhumita Das Biochemistry Lab, GNRC Hospitals, Dispur, Guwahati, Assam

Objective: The present study was conducted to analyze (p = 0.001). Another interesting finding of this study is that and correlate the plasma homocysteine level with the the mean folate (5.3ng/ml & 8.4ng/ml respectively; p=0.000) blood levels of folic acid, vitamin B6 and vitamin B12, and mean vitamin B12 (249pg/ml & 423pg/ml respectively; as decreased circulating concentrations of folate, vitamin p=0.000) are significantly low in the group of subjects B-12, and pyridoxine are associated with increased serum having higher homocysteine level than the subjects having Homocysteine concentration. normal homocysteine suggesting that decreased circulating concentrations of folic acid and vitamin B12 are associated Materials & Method: A total of 970 healthy volunteers with hyperhomocysteinemia. But Vitamin B6 deficiency is of both genders in the age group of 35-86 years, were not associated with increased serum homocysteine con- enrolled in the study. Plasma total homocysteine and centration (p=0.169) in our study. Vitamin B6 were determined by HPLC (High Performance Liquid Chromatography) based on fluorimetric detection. CONCLUSION: To our knowledge, this is the first study Folic acid and Vitamin B12 were estimated by Chemilumi- to report data on Homocysteine concentrations in entire nescent Immunoassay. North East and to evaluate the associations of vitamin deficiency with the risk of hyperhomocysteinemia. The Results: 55% of the community shows hyperhomocysteine- findings of this population based study suggest that the mia with a mean of 18.41μmol/L. Males (mean=20.36μmol/L) prevalence of hyperhomocysteinemia is high in this popu- have a greater tendency to have higher homocysteine than lation sample. the females (mean=16.37μmol/L), which is highly significant

HOSPITAL BASED STUDY ON REFERENCE RANGE OF SERUM PSA LEVEL IN HEALTHY INDIAN MALES

Aditi Gupta, Deepa Gupta, Kamini Vinayak, Vijay, NP Gupta, Vijay Kher, Arun Raizada Medanta-The Medicity Gurgaon Objective: Prostate cancer (PC) is a non dermatological were performed in clinical chemistry analyzer, based on malignancy in males. PSA has been widely used as an initial Immunometric Analyzer Technique (Chemiluminescence). diagnostic marker for prostate disorder. As there is low The exclusion criteria were patient having any urologi- incidence of prostate cancer in Indian males as compared cal complication, prehistorically issues of prostate gland, to western countries males, there are very few records prostate surgery, chemotherapy or radiotherapy. Patients available that indicates routine use of PSA for early de- with pus cells more than 4 were also excluded. tection of PC. In India the reference value of PSA in use Result: In our present study the mean of serum PSA is reference value of western countries. Previous studies range is 1.35±1.9 and variance is 3.7 in overall subject. The suggest that PSA range varies from countries to countries minimum and maximum range is 0.06 and 25.9. The 95% with age, race and religion. Thus the reliable cutoff value percentile range of serum PSA is 3.08 of PSA in Indian males is in debate. This study has been carried out to keep this point in mind that the need to Conclusion: Serum PSA range is more valuable to find create reference interval of PSA in Healthy Indian males. out prostate disorder in patients. The above data suggest that the reference range of serum PSA is 3.08 ng/ml in Method: This was a Descriptive case study on 1283 patients Indian males. enrolled in Executive Health Checkup in Medanta Hospital, Gurgaon from Jan 2010 to Jan 2012. All serum PSA tests

123 Ind J Clin Biochem S109

STUDY OF GLUCOSE 6PHOSPHATE DEHYDROGENASE DEFICIENCY IN TRIBAL AND NON TRIBAL POPULATION OF JHARKHAND

Dr. Shashi Bala Nag, Dr. (Prof.) K.K. Sinha Department of Biochemistry, Rajendra Institute of Medical Sciences, Ranchi, Jharkhand Objective: The incidence of the disease in the patients Result: In this study group of 137 subjects, 11.68% (16) admitted in RIMS and those attending OPDs were studied were G6PD deficient, (14 were male and 2 female). Among by analyzing the G6PD activity. Jharkhand is a malaria non-tribal subjects 10.71% (9) and 13.20% (7) of tribal were endemic zone. Other infections are also common here G6PD deficient. Hemoglobin level in the deficient was and estimation of G6PD activity is not common practice subnormal for the age and sex of individual. here. Exposure to precipitating factors can be avoided by Conclusion: In Jharkhand 28% of the total population detection of G6PD deficiency in a person. consists of Tribes and rest 72% by others. Study results Materials and Methods: The study group comprises of 137 suggest G6PD deficiency is not uncommon in both groups subjects. 84 were non-tribal and 53 were tribal from both so it is necessary to carry out G6PD estimation in whole sex and different age group. Qualitative estimation of G6PD population to prevent G6PD deficiency related health activity was done by Methemoglobin reduction test and problems by close monitoring and judicious use of drugs Quantitative estimation by Kornberg & Horecker method, in the deficient subjects. serum bilirubin by Jendrassik and Grof and hemoglobin by Cyanmethemoglobin method.

BIOCHEMICAL PARAMETERS IN DENGUE

Moushumi Lodh1, Debakanta Pradhan2, Shreekant Bharti3, Mridushri Bharti4 1M.D. Biochemistry, 2M.D. Microbiology, 3M.D. Pathology, The Mission Hospital, Durgapur. 4M.B.B.S. PG (Dip) Microbiology, The Mission Hospital, Durgapur. Objective: The acute phase of dengue begins with fever Result: Abnormal levels of aspartate aminotransferase that is indistinguishable from the initial phase of other (AST), alanine aminotransferase (ALT), bilirubin, alkaline acute febrile infectious diseases. Thus, acute dengue in- phosphatase, protein, albumin, gamma-glutamyl trans- fection is often unrecognized until the appearance of the peptidase (G-GT), LDH, ADA, amylase, lipase, creatineph- more severe forms of the disease. This observation leads osphokinase, hsCRP, calcium were observed in most of to underestimation of the actual incidence, as well as in- the patients. Initially, the level of AST was greater than adequate or late treatment of a disabling and potentially that of ALT, increasing to maximum levels ten days after lethal medical condition. The need arises for a detailed the onset of symptoms, then decreasing to normal levels evaluation of potential biochemical indicators in early within 15 days. Of 3 fatal cases, 2 died of dengue shock stages of dengue. We look at the utility of biochemical syndrome and one of liver failure. alterations for timely identification of such patients. Conclusion: It is concluded that dengue fever may cause Method: The impact of dengue on biochemical parameters hepatic injury and in epidemic or endemic areas, dengue was studied on 200 consecutive patients diagnosed with fever infection should be considered in the differential this disease during July to September 2012, in a tertiary diagnosis of hepatitis. Early alterations of biochemical pa- care hospital of Durgapur in West Bengal. rameters like CK, hsCRP, LDH or ADA in fever could also predict dengue. The biochemical basis of the alterations in Dengue were discussed.

123 S110 Ind J Clin Biochem

POSSIBLE MECHANISMS OF MODULATION OF CHOLESTEROL METABOLISM BY MEDICINAL PLANTS IN VITRO AND EX VIVO ASSAYS

Vanitha Reddy and Asna Urooj Department of Studies in Food Science & Nutrition, University of Mysore, Mysore

Elevated plasma cholesterol concentration is a well known drug, statins. Compared to statins, all 3 plants exhibited risk factor for the development of atherosclerosis and coro- higher inhibition of HMG CoA. The Coenzyme A released nary heart disease (CHD). Reductions in plasma cholesterol at the end of the assay were as follows: control -119.70 can be achieved by inhibition of HMG Co A reductase (key nM and statins 94.49nM, CS -72.97nM, CP 55.41nM and enzyme in cholesterol metabolism) and by promoting bile AP 50.23nM. The in vitro bile acid binding capacity was acid binding. The dietary fiber and saponins are known to analyzed using a bile acid mixture similar to that found reduce elevated cholesterol levels by forming complexes in human bile at duodenal physiological pH and compared with cholesterol and bile acids. Medicinal plants are well with cholestyramine- a cholesterol lowering drug. The bile known in reducing the high cholesterol level at biological acid binding capacity of the 3 samples was dose dependent. level. In the present study, three medicinal plants namely Relative to cholestyramine, Canthium (76%) and Costus Abrus precatorius (AP), Canthium parviflorum (CP) and Cos- (74%), being rich in dietary fiber and saponins, exhibited tus speciosus (CS) were studied for their ability to inhibit higher bile acid retardation index (BARI). Results suggest HMG Co A reductase based on the stoichiometric formation that there is scope to explore these medicinal plants as of Coenzyme A (SH) during the reduction of microsomal natural cholesterol lowering agents. Further studies using HMG COA to mevalonate and compared with the standard animal models are underway.

BIOCHEMICAL STUDY OF RISK FACTORS IN MALE PATIENTS NEPHROLITHIASIS

*Kanchan Singh, **Osman Musa, *Farzana Mehdi, *Aparna Misra, *Ajanta Roy *Department of biochemistry, **Department of Surgery, Ears Lucknow Medical College & Hospital, Sarfaraz Ganj, Hardon Road, Lucknow

Introduction: Kidney stone disease is mainly a multi-factorial for routine and microscopy and 24 hrs urine for calcium, disorder resulting from the combined influence of environ- oxalate. phosphate, uric acid, magnesium, citrate, THP mental, biochemical and genetic risk factors Calcium oxalate (tamm horsfall protein). All parameters were checked and phosphate stones are more common In men; average by standard method. Stone analysis was also done by age of onset is third to fourth Wade of life other types standard method. of stones are uric acid, struvite etc. Urine contain various Results: Although nephrolithiasis has multifactorial causa- inhibitors and promoters for stone formation. tion but biochemical constituents of blood and urine like Aim of study: to evaluate the biochemical risk factors in hypercalcaemia, hyperuricemia, hyperphosphetmia, hyper- blood and urine in male patient with nephrolithiasis. parathyroidism has significant association in formation of nephrolithiasis. Promoters of stone formation was raised Material and Method: This study was carried out on a in urine and inhibitor are deranged. oxalate stones. In total of 60 individuals, in which 30 were male patients Conclusion - Our study shows that commonest type of and 30 were healthy male control, attending OPD of Gen- stones are calcium urine promoters of stone formation eral Surgery, Era’s Lucknow Medical College & Hospital, are raised as compared to inhibitors Lucknow. Serum calcium, phosphate, uric acid creatinine, PTH was measured along with analysis of fasting urine Keywords: nephrolithiasis, promoters, inhibitors.

123 Ind J Clin Biochem S111

FETUIN A A NEW TOOL FOR PREDICTION OF PRE DIABETES

Dr. Mridula Mahajan, Mrs. Sukhraj Kaur and Dr. B.S. Bal Department of Biochemistry and Medicine Government Medical College Amritsar, Punjab

Type 2 diabetes mellitus is predeceased by a long asymp- individuals were offsprings of diabetics (either mother or tomatic period designated as pre-diabetes, impaired fasting father or both) and were siblings amongst themselves glucose or insulin resistance. This asymptomatic period if belonging to age group of 10-55 years. Various biochemi- diagnosed at the right time can help the individuals to lead cal investigations such as fasting plasma glucose, glycated a normal healthy life and relieve the society as a whole Hb, S. insulin, c-peptide, complete lipid profile and fetuin-a, of the burden of treatment of diabetes mellitus. Keeping apart from anthropometric measurements were carried out. in view the magnitude of incidence of pre-diabetes, the It was observed that fetuin-a had a positive correlation present study was planned to study fetuin- a, and find its with the various biochemical investigations thus suggesting correlation if any with various biochemical investigations that fasting plasma glucose and fetuin-a can be used as used as markers of pre-diabetes or insulin resistance. To a tool to determine the susceptibility of an individual to collect the sample size a door to door survey of some develop pre-diabetes and thus diabetes mellitus. of the villages of border belt of Amritsar and Tarn Taran Keywords: insulin resistance, pre diabetes, fetuin-a, fam- district was conducted. A total of 400 families compris- ily history ing of 1161 individuals were recruited for the study. These

THE ROLE OF SERUM PROCALCITONIN AND CREACTIVEPROTEIN IN UPPER RESPIRATORY TRACT INFECTIONSAS A PRIMARY DIAGNOSTIC MARKER

P. Chauhan*, P. Dabla**, A. Sood* Deptt. of Biochemistry, E.S.I. Dental College & Hospital, Rohini, Delhi* DepttOf Biochemistry, C.N.B.C, Affiliated to MAMC, Geeta Colony, Delhi**

Objective: Upper Respiratory Tract infections (URI’s) are blood cells (WBC) counts and microbiological culture. the most frequent of the community with major social Result: In 40 patients with acute tonsillitis the level of and financial impact. Thus, early and proper diagnosis is PCT was highly significant (p<0.05). PCT levels in acute of utmost importance. Biomarkers such as the circulating tonsillitis showed significant correlation with CRP (P<0.050) levels of Procalcitonin (PCT) have been shown to be el- and with TLC (p< 0.01) whereas in rhinosinusitis cases CRP evated in systemic bacterial infections, but remain relatively showed significant correalation with WBC (p< 0.05). On low in viral infections and inflammatory diseases and have the other hand serum CRP levels strongly correlated with been suggested as signals for the initiation of antimicrobial body temperature and WBC. Patients with acute tonsillitis therapy. Thus, purpose of our study is to assess the role showed low CRP as compared to rhinosinusitis. of PCT and C-reactive Protein (CRP) as a inflammatory markers in patients with URI’s for the purpose of early Conclusion: These results suggest that both PCT and CRP diagnoses and intervention. found to be good indicator of bacteremia. Further inves- tigation and research is needed in procalcitonin guided Methods: The study design comprised ofFifty patients with antibiotic treatment in upper respiratory tract infection bacterial URI’s. It includes 32 male and 18 female with a which is required to combat antibiotic resistance without median age of 16-48 years. The PCT was measured by using jeopardizing the safety of patients.Therefore CRP and PCT Immunochromatograficsemiquatitative test (BRAHMS PCT-Q can be used as a marker to predict the severity of URI’s test), as bacterial inflammation marker. It was compared especially in acute tonsilitis with other markers, such as C-Reactive Protein (CRP), white

123 S112 Ind J Clin Biochem

FIRSTPHASE INSULIN SECRETION, INSULIN SENSITIVITY, GHRELIN, GLP1 GLUCAGON LIKE PEPTIDE1, AND PYY PEPTIDE TYROSIN TYROSIN CHANGES 72 H AFTER SLEEVE GASTRECTOMY IN OBESE DIABETIC PATIENTS: THE GASTRIC HYPOTHESIS

Dr Mukesh Meena, Dr M K S Sheikh, Dr Mohit Bhandari, Dr Meena Varma, Dr R.K. Shrivastav Dr Shilpa Mittal

Aim & Objective: The aim of this study is to evaluate the decrease in ghrelin values and an increase in GLP-1 and possible role of sleeve GASTRECTOMY (SG) per se in the PYY values. However, in the group of patients with disease reversibility of diabetes. duration >10.5 years, the differences were not significant except for the late insulin AUC. Postoperative basal and Methods: Insulin secretion and peripheral insulin sensitiv- intravenous glucose-stimulated ghrelin levels were lower ity using the intravenous glucose tolerance test (IVGTT) than preoperative levels in all groups of patients. Basal and were assessed in 100 obese type 2 diabetic patients and intravenous stimulated GLP-1 and PYY postoperative values in 50 non-diabetic obese patients before and 3 days after were higher than preoperative levels in all groups. SG, before any food intake and any weight change occur- rence. At the same time, ghrelin, GLP-1, and PYY levels Conclusions: Restoration of the first phase of insulin se- were determined. cretion and improved insulin sensitivity in diabetic obese patients immediately after SG, before any food passage Results: In diabetic patients who had the disease less than through the gastrointestinal tract and before any weight 10.5 years, the first phase of insulin secretion promptly loss, seem to be related to ghrelin, GLP-1, and PYY hor- improved after SG. The early insulin area under the curve monal changes of possible gastric origin and was neither (AUC) significantly increased at the postoperative IVGTT, in- meal- nor weight-change-related. Duration of the disease dicating an increased glucose-induced insulin secretion. The up to 10.5 years seems to be a major cut off in the second phase of insulin secretion (late AUC) significantly pathophysiological changes induced by SG. A “gastric” decreased after SG in all groups, indicating an improved hypothesis may be put forward to explain the anti-diabetes insulin peripheral sensitivity. In all groups, pre- and post- effect of SG. operatively, intravenous glucose stimulation determined a

GLYCATED ALBUMIN AN INDICATOR OF GLYCEMIC CONTROL

Brijesh Mukherjee

The progressive complications of unmanaged diabetes complications. Diabetes monitoring for protein glycation, include heart disease, blindness, kidney failure, amputa- an essential element for the long-term control of the tion of extremities due to circulation problems, and nerve complications of diabetes mellitus, is currently managed disorders, as well as other chronic conditions. Decades by a combination of daily self-monitoring of blood glucose of research have established that prolonged exposure to (SMBG) measurements and physician-assessed hemoglobin excess glucose is the cause of diabetes complications, and A1c (A1C) levels every 3–6 months. Short term methods like that long-term control of blood glucose levels is required self monitoring of blood glucose and long term methods to avoid or lessen the damage caused by excess glucose. like measurement of HbA1c have limitations. Various re- The process of protein glycation is now understood to be searchers have identified glycated albumin (GA) as the ideal both a marker for the progress of diabetes complications marker for an intermediate index to measure glycation. and an underlying cause of many of the most serious

123 Ind J Clin Biochem S113

ABDOMINAL FAT DISTRIBUTION, INSULIN RESISTANCE AND CARDIOVASCULAR RISK PROFILES IN FEMALES WITH POLYCYSTIC OVARIAN DISEASE ATTENDING OBSTETRICS AND GYNAECOLOGY DEPARTMENT T. RIMS

Vanita Lall1, Chandrahaas Prasad2, Archana Kumari3 1Associate Professor, Department of Biochemistry, AIIMS, Jodhpur, 2Associate Prof. Department of Pathology 3Assistant Professor in Department of Obstetrics and Gynaecology at RIMS, Ranchi

Aim: to determine the association of abdominal fat distribu- Result: Mean age of the subjects were 25.98 years. Eighty tion with insulin resistance (HOMA IR) and cardiovascular females were obese and 43 had android obesity. Insulin risk of females with PCOS. resistance (HOMA-IR) was present in 67 women and 56 women had cardiovascular risk. (>1). Women with waist Method: Cross sectional study that included detailed hip ratio >0.85 were more likely to be insulin resistant clinical examination, BMI, waist hip ratio, insulin resis- and at an increased risk of cardiovascular events. tance (HOMA –IR) and cardiovascular risk scores for 100 women between January 2008-Dec 2009 were calculated. Conclusion: Long term health hazards of PCOS have to be Biochemical Parameters included Blood sugar (fasting), considered as these may be prevented by early identifica- serum insulin levels, triglycerides, total cholesterol, HDL tion and intervention, such as changing the lifestyle of the Cholesterol, LDL and VLDL. individual by execise, food habits etc.

NEW HORIZONS IN DIAGNOSTIC EFFICACY OF PERIODONTAL DISEASES

Kaur Ramanjit1, Ghalaut Veena S.1, Ghalaut Pankaj2, Bansal Piyush1, Manjubala1, Manjulata Kumawat1, Kiran Dahiya1 1Dept. of Biochemistry, PGIMS, Rohtak, Haryana, 2Dept. of Periodontics, Govt. Dental College Rohtak, Haryana

Study of changes in the enzymatic activity in saliva due Thermoking (Konelab) automated analyzer. Kinetic methods to pathological and metabolic changes in the gingiva and was used for the determination of AST, ALT, ALP and LDH periodontium in response of an organism to the periodontal activity. Fluoride was measured using Orion Ion Selective infection may help in accurate assessment of the pathology Electrode from thermo- scientific. Statistical analysis was and provide a non-invasive and convenient diagnostic tool. done using microsoft excel and Student’s t-test was ap- Fluoride has been considered a protective element, but it plied to compare the study and control group. may itself be involved in pathogenesis and its levels may In the saliva, from the patients suffering with periodon- serve as a marker of periodontal pathology. tal disease, there was a significant increase in activity We estimated intracellular enzymes and fluoride levels in of LDH, AST, ALT and ALP in comparison to the control 30 healthy adults ( volunteers ) of both sexes in the of group. Fluoride levels were also significantly raised in sa- age group of 18 – 40 years and in 30 persons, of both liva of the patients with periodontal disease. The results sexes, aged 18 – 40 years, with periodontal disease. Pa- of the study emphasize the use of salivary enzymes and tients with a probing depth >5 mm, bleeding on probing salivary fluoride as objective and simple markers of the and alveolar bone loss >40% were included. pathological state of the periodontal tissues which opens new horizons in diagnostic efficacy. 3.0 mL of un-stimulated mixed saliva was collected in a sterile test tube. Samples were then centrifuged at 10000 Keywords: Sialochemistry, Intracellular enzymes, Fluoride, rpm for 10 min and processed on the same day of col- periodontitis lection. Salivary enzyme activities were measured on a

123 S114 Ind J Clin Biochem

IODINE DEFICIENCY DISORDERS IN & AROUND MEERUT CITY

Suryakant Nagtilak, Naved Ahmad, Amit Rastogi*, Pawan Parashar** Department of Biochemistry, Medicine (Endocrinology)*, Community Medicine**, Subharti Medical College, Meerut

Introduction: Thyroid dysfunctions are the most common & TSH by ELISA & Iodine content in salt, analysed using form of IDD in India, it may be due qualitative or quantita- standard titration. tive alterations in hormonal secretion, the increase in size Results: The study reveals hypothyroidism 17%, hyperthy- of the gland or both mechanisms. The shortage of thyroid roidism 5%, euthyroidism 78% respectively, iodine content hormones produces hypothyroidism and excessive secretion in edible salt revealed that in RHTC 96% house holds causes hyperthyroidism. The dysfunctions manifest in wide consuming iodised, 4% using non iodised salt & 42% house s p e c t r u m o f c l i n i c a l a n d b i o c h e m i c a l d i s e a s e . holds consuming salt < 15 ppm Iodine. In UHTC 100% Objectives: Estimate (T3, T4) and TSH levels in patients house holds using iodised salt but around 4% using < 15 attending medical [Endo] OPD/IPD, salt analysis for content ppm iodine. of iodine & consumption pattern in study area. Conclusion: This is for the first time clinico-Biochemical Materials & Methods: Study was carried in & around Study conducted on IDD in this region. Study highlights [RHTC & UHTC] Meerut city in 504 patients attending that hypothyroidism, was alarmingly high with female Medical O.P.D/ I.P.D Specialty-Endocrinology assessed for vulnerability with a need to monitor iodised salt distribu- thyroid hormone profile & TSH aged between 15 to 60 tion/storage/consumption practice by health authorities yrs of both sexes were screened. Fasting venous blood & to educate, convince both consumers and salt traders samples were collected & assayed for thyroid hormones of rural areas about health benefits of adequate use of iodised salt.

123