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click here for freelancing tutoring sitesRAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA BANGALOREANNEXURE-II

PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION

1. Name of the candidate and address (in block letters)

Dr. RASHMI SINHADept of Biochemistry,M.R Medical College,Gulbarga– 585105

2. Name of the Institution H.K.E Society’sM.R Medical college,Gulbarga-585105

3. Course of Study and subject M.D. (Biochemistry)

4. Date of Admission of Course 28-4- 2009

5. Title of the topic “STUDY OF SERUM & URINE PROTEIN THIOLS IN ESSENTIAL HYPERTENSION”

6. Brief Resume of the Intended work6.1. Need for the Study: Essential hypertension is common in developed

societies & is a major risk factor for cardiovascular disease. It is likely to be the one of the consequences of an interaction between environmental & genetic factors.1

Several previous studies have indicated that

essential hypertension is a state of increased oxidative stress with increased reactive oxygen species (ROS) & imbalance of ROS & the antioxidant defense mechanism.2,3

The thiol group on proteins has an important role in antioxidant status of the body. This group of serum proteins especially albumin has been suggested to play an important role as antioxidant in plasma & extravascular spaces4. Several investigators suggested the enhanced thiol group’s oxidation involved in the pathogenesis of renovascular hypertension.5,6,7 However, the information regarding oxidative protein damage in patients with essential hypertension have not been studied exclusively & there is paucity of information in the literature in this regard.

As it is well known fact that about 6% to 40% of uncontrolled hypertension is associated with proteinuria & albumin is a major protein excreted in urine of patient.8,9,10 The current study is designed to establish the relationship between serum albumin & protein thiols with protein – bound thiols in the urine of essential hypertensive patients.

6.2. Review of Literature: In humans, hypertension is also considered as a state of oxidative stress that can contribute to the development of atherosclerosis.11

Previous studies promoted the view that essential hypertension is a state of increased oxidative stress and the data on oxidative protein modification is lacking. 2

Available evidences supports a role of protein thiol groups in this process as they represent a well known target reactive species12,13

Indrajit Sinha et al 14 found that markers of oxidative stress correlated significantly with proteinuria in nephrotic syndrome patients, which is in accordance to the previous studies.

Among hypertensive subjects, patients with microalbuminuria have increased risk of cardiovascular complications.15,16

6.3. Objective of the study: The present study aims to assess the changes in the serum & urine protein thiol level in essential hypertensive patients.

7.Material and methods

7.1 Source of Data:The study would be undertaken at Basaveshwar Teaching and General Hospital attached to M.R.Medical College, Gulbarga.

Present study consists of 50 clinically diagnosed cases of essential hypertension in the department of Medicine during the period of Dec 2009 to June 2011.

Controls would consist of non-hypertensive persons who will be carefully selected & examined in detail with age & sex matched.

7.2 Methods of collection data: (including sampling procedure if any)

Under aseptic conditions, blood samples will be drawn into plain vacutainers from the antecubital vein .The collected blood will be allowed to clot & then centrifuge at 2000rpm for 15 minute for clear separation of serum.

From the same cases & controls, 24hr urine samples will be collected in bottles containing toluene & will be then assayed for urinary proteins & protein thiols.

Methods- Serum & Urine protein thiols will be measured by Spectrophotometric method using 5,5`dithio-bis2-nitrobenzoicacid(DTNB),Serum creatinine, serum albumin, urinary total proteins and creatinine levels will be estimated by spectrophotometric methods using an Semiautoanalyser (ERBA-CHEM 7).

Inclusion Criteria:Cases clinically diagnosed as essential hypertension in department of Medicine at Basaveshwar Teaching & General Hospital, attached to M.R.Medical College, Gulbarga.

Exclusion Criteria:Cases of essential hypertension associated with Diabetes, obesity, Smoking and other types of Hypertension.

7.3 DDoes the study required any investigation & intervention to be conducted on patients or other humans or animals? If so, please describe briefly.

The present study requires investigation to conduct on humans as per ethical guidelines. Volunteer written informed consent will be obtained from all the patients & controls of the study.

7.4 Has ethical clearance been obtained from your institution in case of 7.3

Yes, Ethical clearance has been obtained from the ethical committee of the institution for the study.

8. List of References

1.Harrison’s Principles of Internal Medicine: 17th edition pg.no.- 1554.

2.SimicDV,Mimic-OkaJ,Pljesa-Ercegovac,Savic-adojevic,Opacic M,MaticD,Ivanovic B and Simic. Journal of human hypertension.8 Dec 2005;doi:10.1038/sj.jhh.1001945.

3.Stocker&Keaney.Role of oxidative modifications in Atherosclerosis.PhysiolRev 2003; 84:1381- 1478.

4.HalliwellB;Antioxidants&human disease; a general introduction,Nutr Rev1997,55,544-552. 5.Inagi R, Miyata T .Oxidative protein damage with carbohydrates & lipids in uremia:’Carbonyl stress’Blood Purif 1999; 17:95-98.

6.Himmelfarb J,McMonagle E,McMenamin E .Plasma protein thiol oxidation & carbonyl formation in chronic renal failure. Kidney Int 2000; 58:2571-2578.

7. Mimic Oka J ,Simic T, Pljesa M, Stupar N, Turkovic S. Oxidative modifications of plasma proteins in different stages of chronic renal failure.Facta Univarsitalis 2001;8:1-5.

8.G.Crippa.Microalbuminuria in essential hypertension: Journalof Human Hypertension 2002 ;16 :S74 – S77. 9.Rosa,Torres Tania;Palatini,Paolo.Clinical value of microalbuminaria in hypertention.Journal of human Hypertention, June-2000-Volume18-Issue6 p 645-654. 10.Barry M Brenner,The Kidney,8th edition, page no. 1473-1474. 11. Romero JC, Reckelhoff JF. Role of angiotensin and oxidative stress in essential hypertension. Hypertension. 1999; 34:943–949.

12.Lohose DL, Denu JM, Santoro N and Dixon JI: Role of aspartic acid-181 and Serine -222 in intermediate formation and hydrolysis of mammalian protein-tyrosine-phosphatase PTPI.Biochemistry36:4568-4575, 1997.

13.Zhang ZYand Dixon JE:Active site labeling of the Yersinia protein tyrosine phosphatase the determination of the pKa of the active site cysteine and the function of the conserved histidine 402;Biochemistry 32:9340-9345.

14.SinhaIndrajit,GhoshSandip,Prasenjit Dey,Jose Jacob andDibyajyoti Banerjee. Reduction of urinary thiols in nephrotic syndrome- a possible effect of free iron: Clinica chimica Acta; Volume 355, issues 1 – 2: May 2005.

15.Redon J et al.Factors related to the presence of microalbuminuria in essential hypertention.Am J Hypertens 1994; 801-807.

16. Pedrinelli P et al.Microalbuminuria and endothelial dysfunction in essential hypertention.Lancet 1994;344:14-18.

9. Signature of the Candidate

10. Remarks of the Guide

Essential hypertension is a state of oxidative stress. Because of such oxidative environment, it could lead to decrease in serum and urine protein thiols.The present study is taken up to establish the changes during essential hypertension.Recommended & Forwarded for kind acceptance.

11.Name and Designation of (in block letters)

11.1. Guide

Dr. JAGADISH B. INGIN, M.D.

PROFESSOR & HEADDEPT. OF BIOCHEMISTRY,MAHADEVAPPA RAMPURE MEDICAL COLLEGE, GULBARGA.

11.2. Signature

11.3. Co-Guide

Dr. BASAVARAJ MANGASHETTY M.D ASSOCIATE PROFESSOR DEPT OF MEDICINE ,MAHADEVAPPA RAMPURE MEDICAL COLLEGE, GULBARGA .

11.4. Signature

11.5. Head of the Department

Dr. JAGADISH B. INGIN M.D.

PROFESSOR & HEADDEPT. OF BIOCHEMISTRY,MAHADEVAPPA RAMPURE MEDICAL COLLEGE, GULBARGA

11.6. Signature

12.12.1. Remarks of the Chairmanand Principal

12.2. Signature