RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA
BANGALORE
ANNEXURE II
PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION
|1. |Name of the Candidate |Dr. NALINAKSHI. S |
| |and Address |POST GRADUATE STUDENT, |
| |(in block letters) |DEPARTMENT OF BIOCHEMISTRY, |
| | |J J M MEDICAL COLLEGE, |
| | |DAVANGERE-577 004. |
|2. |Name of the Institution |J J M MEDICAL COLLEGE, |
| | |DAVANGERE – 577 004. |
|3. |Course of study and subject |M.D. – BIOCHEMISTRY |
|4. |Date of Admission to course |26-07-2010 |
|5. |Title of the Topic |“EVALUATION OF SERUM SUPEROXIDE DISMUTASE, PARAOXANASE, |
| | |MALONDIALDEHYDE AND HIGH SENSITIVITY C-REACTIVE PROTEIN IN ISCHEMIC |
| | |STROKE PATIENTS WITH AND WITHOUT TYPE II DIABETES MELLITUS’’. |
|6. |Brief Resume of the intended work : |
| | |
| |6.1 Need for the study: |
| |Stroke is defined as rapidly developed clinical signs of focal disturbance of cerebral function; lasting more than 24hrs |
| |or leading to death, with no apparent cause other than vascular origin1. |
| |Stroke is the third leading cause of death in developed countries. It ranked as the sixth leading cause of Disability |
| |Adjusted Life Years (DALY; one DALY being equal to one year of health lost due to disease) in 1990 and is projected to |
| |rank fourth by the year 20202. |
| |WHO collaborative study in 12 countries showed stroke incidence rates ranged from 0.2 to 2.5/1000 population per year. The|
| |prevalence rate of stroke in India is about 1.54/1000 and death rate about 0.6/1000. It accounts for 10-12% of total |
| |deaths1. The best strategy for reducing its burden is prevention by targeting population at risk for primary prevention |
| |and risk factor management to prevent its recurrence2. |
| |Strokes are currently classified as either ischemic or hemorrhagic. Ischemic stroke occurs when the blood supply to a part|
| |of the brain is suddenly interrupted by occlusion3. |
| |Diabetes is a strong risk factor for ischemic stroke, because of accompanying risk factors, such as obesity and high blood|
| |pressure. Also insulin resistance is implicated as an independent risk factor in the development of atherosclerosis and |
| |stroke3. |
| |Oxidative stress is probably one of the mechanisms involved in neuronal damage induced by ischemia-reperfusion. During |
| |ischemia, high amounts of free radical formation together with reduced antioxidant defense causes oxidative stress that |
| |may play a role in the pathogenesis of stroke-associated neuronal injury4. Brain is very rich in polyunsaturated fatty |
| |acids which are highly susceptible to free radical attack3. |
| |Free radicals that are formed induce lipid peroxidation of polyunsaturated fatty acids and damage biological membranes |
| |producing malondialdehyde (MDA) as one of the end product of lipid peroxidation. The pathophysiological processes involve |
| |pathological permeability of the blood-brain barrier, energy failure, loss of cell ion homeostasis, acidosis, increased |
| |intracellular calcium, apoptosis and neuronal death5. |
| |Antioxidants are exogenous or endogenous compounds acting in several ways, scavenging reactive oxygen species or their |
| |precursors, inhibiting reactive oxygen species (ROS) formation and binding metal ions needed for catalysis of ROS |
| |generation3. |
| |Enzymatic antioxidants like glutathione peroxidase, glutathione reductase, superoxide dismutase, catalase and |
| |non-enzymatic antioxidants like ascorbic acid, tocopherols, β-carotene, bilirubin, uric acid scavenge the free radicals |
| |and protect the cells. Superoxide dismutase (SOD), present in cytosol functions mainly as a first order antioxidant enzyme|
| |by neutralizing the effect of superoxide anion which is an important precursor for oxidative stress. It dismutates |
| |superoxide anion to H2O26,9. |
| |Human serum paraoxanase or PON (aryldialkyl phosphatase) is a protein synthesized in the liver and has antiatherogenic |
| |activity due to its protective function against oxidative modification of low density lipoprotein (LDL)7. |
| |High sensitivity C-Reactive Protein (hs-CRP) is an acute-phase reactant and emerging as a new marker for coronary artery |
| |disease and associated cerebrovascular disorders. Inflammation theory of atherosclerosis suggests – CRP is not only a |
| |marker of inflammation but also proinflammatory, proatherosclerotic and prothrombotic in nature. It enhances production of|
| |MCP-1 (Monocyte chemoattractant protein) recruiting more monocytes, promoting uptake of LDL by macrophages and activation |
| |and thus plaque formation leading to atherosclerosis and subsequently stroke2. |
| |Inflammatory course of atherosclerosis is more severe in diabetics than non-diabetics. Serum CRP levels increase with the |
| |stage of beta cell dysfunction and insulin resistance. CRP induces upregulation of NADPH oxidase and uncoupling of |
| |endothelial nitric oxide synthase (eNOS) in blood vessels of diabetics leading to endothelial dysfunction and excessive |
| |superoxide anion (O2.-) production causing decreased NO (nitric oxide) bioavailability which has an important role in |
| |homeostatic vasodilatation and regulation of blood flow8. |
| |High CRP levels have poor prognostic significance in ischemic stroke patients. Statins or fibrate therapy may be used |
| |which are helpful in controlling, both dyslipidemia and elevated CRP levels. Monitoring the levels of CRP may help in the |
| |prevention and treatment of stroke2. |
| |Human studies on stroke and oxidative stress are limited and controversial. Hence the present study is undertaken to |
| |evaluate by measuring the serum levels of superoxide dismutase, paraoxanase as antioxidant enzymes, hs-CRP as inflammatory|
| |marker and malondialdehyde as lipid peroxidation marker and correlate between serum levels of SOD, PON with serum levels |
| |of hs-CRP and MDA in ischemic stroke patients with and without type II diabetes mellitus. |
| |6.2 Review of literature : |
| |Changes in enzymatic antioxidative defence systems after acute stroke are controversial5. Cherubini et al9 and Demirkaya |
| |et al10 have demonstrated a significantly reduced SOD activity in AIS (acute ischemic stroke) while others have |
| |demonstrated an increased SOD activity11 still others have demonstrated an unchanged SOD activity in AIS4. |
| |Significant reduction in paraoxanase activity was found in ischemic stroke patients with age and oxidized LDL |
| |significantly increased7. |
| |Studies have shown that ischemic stroke is associated with increased oxidative stress reflected by the lipid peroxidation |
| |product malondialdehyde which is seen to be increased and the reduced antioxidant status in ischemic stroke patients with |
| |and without diabetes3. |
| |Demirkaya et al showed that patients with acute ischemic stroke had significantly higher levels of MDA in their red blood |
| |cells compared with controls which also correlated with infarct size and initial stroke severity3, 10. |
| |A meta – analysis of studies with long term follow up showed that the risk for stroke in healthy individuals with the |
| |highest quartile of CRP concentrations increased nearly 1.7 fold compared with those with the lowest quartile2, 12. |
| |The Framingham study, a cohort follow up study stated that independent of other cardiovascular risk factors, elevated |
| |plasma CRP levels significantly predict the risk of future ischemic stroke and TIA (Transient Ischemic Attack) in the |
| |elderly13. |
| |The Hisayama study, a cohort follow up study in middle aged and elderly Japanese men & women found that elevated serum |
| |hs-CRP levels are an independent risk factor for future ischemic stroke in men and that the coexistence of another risk |
| |factor with high hs-CRP levels increased the risk further14. |
| |Admission CRP after acute ischemic stroke is associated with stroke severity and long-term mortality. High CRP levels may |
| |be considered as a “red flag” marker of high mortality15. |
| |6.3 Objectives of the study : |
| |To evaluate the serum levels of superoxide dismutase and paraoxanase as antioxidant enzymes. |
| |To evaluate the serum levels of hs-CRP as inflammatory marker. |
| |To study the serum levels of malondialdehyde as marker of lipid peroxidation. |
| |To evaluate the correlation of the serum levels of SOD and PON with hs-CRP and MDA in ischemic stroke patients with and |
| |without type II diabetes mellitus and in healthy individuals (as control). |
|7. |Materials and methods : |
| |7.1 Source of data : |
| |A cross sectional study will be carried out for a period of one year. The patients will be selected from Chigateri General|
| |Hospital and Bapuji Hospital, Davangere (both hospitals are attached to the teaching institute J.J.M. Medical College, |
| |Davangere). |
| |The study group consists of acute ischemic stroke patients with and without type II diabetes mellitus. The control group |
| |consists of age and sex matched healthy individuals. Both cases and controls will be interviewed to obtain relevant data |
| |after taking informed consent. Based on inclusion and exclusion criteria minimum of 60 cases will be selected. Among 60 |
| |cases approximately 30 cases will be with type II diabetes mellitus and the rest will be without type II diabetes |
| |mellitus. About 30 age and sex matched healthy individuals will be taken as controls. |
| |Inclusion criteria : |
| |Newly diagnosed acute ischemic stroke patients, diagnosed by Computed Tomography scan, clinical signs and symptoms and |
| |biochemical parameters in the age group of 30-80 years admitted within first 48 hours of stroke onset. |
| |Acute ischemic stroke patients with type II diabetes mellitus. |
| |Acute ischemic stroke patients without type II diabetes mellitus. |
| |Age and sex matched healthy individuals without any major illness and not on any medications as controls. |
| |Exclusion criteria : |
| |Patients with recent clinical infection, chronic inflammatory condition, or neoplastic disease. |
| |Patients with surgery or major trauma in the previous month. |
| |Patients with any major renal or hepatic disease. |
| |Patients on antioxidant medications. |
| |Individuals who are chronic smokers or chronic alcoholics. |
| | |
| |7.2 Method of collection of data (including sampling procedure if any): |
| |About 5ml of venous blood will be drawn from subjects (from large peripheral veins) under aseptic precautions in a sterile|
| |bulb. Serum will be separated by centrifugation and used for analysis of MDA, SOD, PON and hs-CRP. |
| |Estimation of serum malondialdehyde (MDA) by Nadiger et al method16. |
| |Principle: Serum is treated with 10% trichloroacetic acid (TCA) followed by addition of 0.67% thiobarbituric acid (TBA) |
| |and to that solution 1ml of distilled water is added. The mixture is centrifuged and clear pale pink supernatent is |
| |collected. Colour intensity is measured at 530nm in a colorimeter. |
| |Estimation of serum superoxide dismutase (SOD) by Marklund and Marklund method6. |
| |Principle: Superoxide anion is involved in auto-oxidation of pyrogallol at alkaline pH (8.5). The superoxide dismutase |
| |inhibits auto-oxidation of pyrogallol, which can be determined as an increase in absorbance at 420nm. |
| |Estimation of serum paraoxanase (PON) as done by Sarkar7. |
| |Principle: PON activity is measured by using 5.5 mmol/l p-nitrophenyl acetate as a substrate. The increase in the |
| |absorbance at 412nm due to formation of p-nitrophenol is measured using spectrophotometer. |
| |Estimation of serum high sensitivity C-Reactive protein (hs-CRP) by Immunoturbidimetry17. |
| |Principle: Turbidimetry measures the reduction in the light transmission caused by particle formation and it quantifies |
| |the residual light transmitted. It is based on the principle of agglutination reaction. The test specimen is mixed with |
| |the activation buffer (R1) and the antihuman antibody solution (R2) and allowed to react. |
| |Presence of hs-CRP in the test specimen forms an insoluble complex producing a turbidity, which is measured at wavelength |
| |600nm. The resulting turbidity corresponds to the concentration of the hs-CRP in the test specimen. |
| |Statistical analysis : |
| |Results will be subjected to appropriate statistical analysis. Unpaired ‘t’ test will be used to compare between two |
| |groups. Correlation analysis will be done using Pearson’s correlation co-efficient. |
| | |
| |7.3 Does the study require any investigations or interventions to be conducted on patients or other humans or animals? If|
| |so, please describe briefly. |
| |Yes |
| |Estimation of malondialdehyde (MDA), superoxide dismutase (SOD), paraoxanase (PON), high sensitivity C-Reactive Protein |
| |(hs-CRP) using human blood sample in ischemic stroke patients with and without type II diabetes mellitus and in healthy |
| |individuals as controls. |
| | |
| |7.4 Has ethical clearance been obtained from your institution in case of 7.3? |
| |Yes |
|8. |List of References : |
| |Park K. Epidemiology of chronic non communicable diseases and conditions. In: Park’s textbook of preventive and social |
| |medicine. 20th ed. Jabalpur: Banarsidas Bhanot; 2009. p. 327,328. |
| |Dhamija RK, Arora S, Gaba P, Jais M, Kaintura A, Kumar M and Bhattacharjee J. Study of genetic, metabolic and inflammatory|
| |risk factors in patients of acute ischemic stroke. IJCB 2008;23(2):136–143. |
| |D’souza B, D’souza V, Sowmya S, Seema G, Hemalatha CN, Anu M et al. A comparative study on oxidative stress and |
| |antioxidant status in ischemic stroke patients with and without diabetes. IJCB 2008;23(3):218-222. |
| |Abdul-Fattah D, Abdel-Ghani AM, El-Azouni OMMA, Eltarhouny SA, Gharieb AF. Acute ischemic stroke: clinical, radiological |
| |and antioxidant profile study. Egypt J. Neurol. Psychiat. Neurosurg. 2006;43(1):159-166. |
| |Aygul R, Kotan D, Demirbas F, Ulvi H and Deniz O. Plasma oxidants and antioxidants in acute ischemic stroke. Journal of |
| |International Medical Research 2006;34:413-18. |
| |Nischal HK, Sharma MP, Goyal RK and Kaushik GG. Serum superoxide dismutase levels in diabetes mellitus with or without |
| |microangiopathic complications. JAPI 1998;46(10):853-55. |
| |Sarkar PD and Rautaray SS. Oxidized LDL and Paraoxanase status in ischemic stroke patients. Indian J Physiol Pharmacol |
| |2008;52(4):403-7. |
| |Khan DA and Qayyum S. Evaluation of cardiac risk by oxidative stress and inflammatory markers in diabetic patients. Pak J |
| |Med Sci 2009;25(5):776-781. |
| |Cherubini A, Polidori MC, Bregnocchi M, Pezzuto S, Cecchetti R, Ingegni T et al. Antioxidant profile and early outcome in |
| |stroke patients. Stroke 2000;31:2295-2300. |
| |Demirkaya S, Topcuoglu MA, Aydin A, Ulas UH, Isimer AI and Vural O. Malondialdehyde, glutathione peroxidase and |
| |superoxide dismutase in peripheral blood erythrocytes of patients with acute cerebral ischemia. European Journal of |
| |Neurology 2001;8(1):43-51. |
| |Kocaturk PA, Akbostanci MC, Isikay C, Ocal A, Tuncel D, Kavas GO et al. Antioxidant status in cerebrovascular accident.|
| |Biol Trace Elem Res 2001;80(2):115-24. |
| |Hsu-Ko Kuo MPH, Dr Chung-Jen Yen, Chia-Hsuin Chang, Chen-Ko Kuo, Jen-Hau Chen and Farzaneh Sorond. Relation of C-reactive |
| |protein to stroke, cognitive disorders, and depression in the general population: systematic review and meta-analysis. The|
| |Lancet Neurology 2005;4(6):371-380. |
| |Rost NS, Wolf PA, Kase CS, Kelly-Hayes M, Silbershatz H, Massaro JM et al. Plasma concentration of C- reactive |
| |protein and risk of ischemic stroke and transient ischemic attack: the Framingham Study. Stroke 2001;32:2575-2579. |
| |Wakugawa Y, Kiyohara Y, Tanizaki Y, Kubo M, Ninomiya T, Hata J et al. C-reactive protein and risk of first-ever ischemic |
| |and hemorrhagic stroke in a general Japanese population: the Hisayama Study. Stroke 2006;37:27-32. |
| |Idicula TT, Brogger J, Naess H, Waje-Andreassen U and Thomassen L. Admission C-reactive protein after acute ischemic |
| |stroke is associated with stroke severity and mortality: The ‘Bergen stroke study’. BMC Neurology 2009;9:18. |
| |Kalavacherla US, Ishaq M, Rao URK, Sachindranath A and Hepsiba T. Malondialdehyde as a sensitive marker of inflammation in|
| |patients with rheumatoid arthritis. JAPI 1994;42(10):775-776. |
| |Hind CRH and Pepys MB. The role of serum C-Reactive protein (CRP) measurement in clinical practice. Int Med |
| |1984;5:112-151. |
|9. |Signature of candidate | |
|10. |Remarks of the guide |Human studies on stroke and its consequent oxidative stress are limited. |
| | |Therefore, the present study is undertaken to evaluate antioxidant status and|
| | |hs-CRP as inflammatory marker and it may be helpful in understanding the use |
| | |of antioxidants as therapeutic agents in the treatment of ischemic stroke. |
|11. |Name & Designation of (in block letters) | |
| |11.1 Guide | |
| | |Dr. P.M. GANGADHARA SWAMYM.Sc., PhD., |
| | |PROFESSOR, |
| | |Department of BIOCHEMISTRY, |
| | |J.J.M. Medical College, |
| | |DAVANGERE - 577 004. |
| |11.2 Signature | |
| | | |
| |11.3 Co-Guide | |
| | |Dr. P.M. VINAY SWAMYM.D., |
| | |PROFESSOR, |
| | |DEPARTMENT OF MEDICINE, |
| | |J.J.M. MEDICAL COLLEGE, |
| | |DAVANGERE - 577 004. |
| |11.4 Signature | |
| | | |
| |11.5 Head of the Department | |
| | |Dr. D.S. JAYAPRAKASH MURTHYM.D., |
| | |Professor and Head, |
| | |Department of BIOCHEMISTRY, |
| | |J.J.M. Medical College, |
| |11.6 Signature |DAVANGERE - 577 004. |
| | | |
| | | |
|12. |12.1 Remarks of the | |
| |Chairman & Principal | |
| | | |
| |12.2 Signature | |
................
................
In order to avoid copyright disputes, this page is only a partial summary.
To fulfill the demand for quickly locating and searching documents.
It is intelligent file search solution for home and business.
Related searches
- archives of biological sciences journal
- definition of social sciences pdf
- blackrock health sciences fact sheet
- list of social sciences degrees
- blackrock health sciences trust
- blackrock health sciences stock
- archives of biological sciences impact
- institute of mathematical sciences chennai
- university of natural health scam
- university of texas health system
- university of tampa health center
- university of medical sciences ondo