Journal of Clinical and Diagnostic Research, ISSN - 0973 - 709X

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MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
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Dr. Mamta Gupta
Consultant
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Aug 2018




Dr. Rajendra Kumar Ghritlaharey

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Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
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Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2010 | Month : August | Volume : 4 | Issue : 4 | Page : 2752 - 2756 Full Version

Oxidative Stress And Antioxidants In CRF Patients Before And After Dialysis


Published: August 1, 2010 | DOI: https://doi.org/10.7860/JCDR/2010/.812
SATISHKUMAR D *, VISHALI V**, INDUMATI V ***, KODLIWADMATH M V ****, DEVERANAVADGI BB *****, CHANDRAKANTH K H******

*(MD) , **(MBBS), ***( MD)Department of Biochemistry, VIMS, Bellary, Karnataka, (India) ****(MD)(Biochemistry) , ****Department of Biochemistry, Navodaya Medical College, Raichur, (India) *****(MD)(Biochemistry),*****(MD)(Biochemistry), Department of Biochemistry Shri B M Patil Medical College, Bijapur,(India)

Correspondence Address :
Dr.SatishKumar.D
Assistant Professor
Department of Biochemistry
VIMS, Bellary-583104
Karnataka,(India)
Email id: drsatishkumard@yahoo.co.in
Phone No: 09632061135

Abstract

Chronic Renal Failure (CRF) is a debilitating condition which is responsible for high morbidity and mortality. Tissue injury due to free radicals is commonly seen in a variety of disease processes. The aim of the present study was to investigate the possible free radical mediated tissue damage in CRF before and after dialysis, by measuring Melondialdehyde (MDA) which is marker of oxidative stress. Antioxidants like Vitamin A, Vitamin E and Vitamin C, which prevent oxidative damage, were also measured. The study included 30 healthy persons as controls and 30 patients of CRF (before dialysis and after dialysis) as the study cases. All parameters were assessed by spectrophotometric methods.
Results: MDA levels were found to be significantly increased in pre dialysis CRF patients as compared to the controls (P< 0.001) and the levels further increased in post dialysis CRF patients as compared to that in pre dialysis CRF patients (P <0.001). The levels of antioxidants like Vitamin A, Vitamin E and Vitamin C were significantly decreased in pre dialysis patients as compared to the controls (P=<0.001) and the values further significantly decreased in post dialysis patients as compared to the pre dialysis CRF patients(P<0.001).
Conclusion: This study indicates that there is considerable oxidative stress in patients with CRF, which is further exacerbated by hemodialysis, as evidenced by increased lipid peroxidation and low antioxidant levels.

Keywords

Antioxidants, Chronic renal failure, Dialysis, Melondialdehyde,

Introduction
Chronic renal failure is a debilitating condition which is responsible for high morbidity and mortality and it is a financial burden on both the government and the society. True data on the incidence and the prevalence of End Stage Renal Disease (ESRD) in India is lacking, because no National Registries exist here. A high incidence of ESRD has also been noted in Asians of Indian origin in the United Kingdom. If the incidence of ESRD is indeed 100 patients/ million population/ year, this would mean  100,000 new patients every year for a population of 1 billion in India (1).

The present study is focused on the role of free radical mediated tissue damage in chronic renal failure (CRF) and dialysis. The sources of free radicals (FR) are activated macrophages, vascular cells and various glomerular cells, including fibroblasts and renal interstitial cells. Different cellular enzymes including mitochondrial oxidases, lipooxygenases, myeloperoxidase, NADPH oxidase, xanthine oxidase and nitric oxide synthase have been identified as the cellular sources for the formation of reactive oxygen species (ROS) (2).

Generation of oxidative free radicals (OFR) is increased in many pathological conditions. The cellular constituents and biomolecules are subject to free radical attack. Polyunsaturated fatty acids (PUFAs) present in cell membranes are readily attacked by oxygen free radicals. The oxidative destruction of polyunsaturated fatty acids is known as lipid peroxidation. It is a destructive chain-reaction that can damage other cellular components by the production of reactive aldehydes (3). Lipid peroxidation has been implicated in a wide range of tissue injuries and diseases (4). Proteins are subject to free radical mediated destruction which may lead to structural loss of the enzymes or enzymatic deactivation. Nucleic acids are prone to base hydroxylation, cross linking or stand scission, which may result in mutation or even cell death (5). Extracellular tissue components including hyaluronic acid and collagen are vulnerable to injury by toxic free radicals (FRs), thus resulting in damage to the architectural integrity of the tissues (6).

The present study was planned to investigate the possible FR mediated injury in CRF patients who were undergoing haemodialysis by measuring the oxidative stress. Oxidative stress is assessed by measuring malondialdehyde (MDA), which is a marker and a product of lipid peroxidation. The study also included the assessment of the non-enzymatic antioxidants Vitamin E, Vitamin A and Vitamin C in CRF patients before and after dialysis, which prevent oxidative damage and scavange or neutralize free radicals which are produced in disease processes.

Material and Methods

Materials
The sample size included 60 subjects, that is, 30 normal healthy controls and 30 CRF patients who were attending the dialysis unit over a period of one year. These 30 CRF patients had a urine output of less than 600ml per day. The average serum creatinine level was 8.2±3.1 mg/dl and the average blood urea level was 159±40 mg/dl. The causes for CRF in these patients were Hypertensive Nephropathy, Obstructive Uropathy, Acute Gastroenteritis, Acute Pyelonephritis and Diabetic Nephropathy. Most of these patients had Microcytic Hypochromic Anaemia. Ultrasonography findings showed grade II-III nephropathic changes.

Patients with reversible renal involvement, advanced form of major extra renal complications like cerebro vascular accidents, coronary artery disease, neoplasia and active infection elsewhere in the body were excluded from the study. The study was approved by the Medical College ethical and research committee

Methods
6 ml of blood was drawn by venipuncture from the controls and the CRF patients who were selected for dialysis (before and immediately after dialysis) and was collected in a heparinized tube (5 units/ml of blood) by using aseptic precautionary measures.

1ml of blood was processed for the estimation of Malondialdehyde by the Thiobarbituric acid method (7) and the rest of the sample was centrifuged and plasma was separated for
the analysis of antioxidants like Vitamin E (-tocopherol) by the Quaife et al. method (8),Vitamin A (Retinol) by the Bessay et al method (9) and Vitamin C (Ascorbic acid) by the Evelyn and Malloy method (10).


Estimation of Malondialdehyde (MDA) (7)
This reaction depends on the formation of a pink coloured complex between malondialdehyde and thiobarbituric acid (TBA), having a maximum absorption at 532 nm.

Estimations of Plasma-Tocopherol(8)
This method is based on the Emmerie Engel reaction. The Xylene extract of plasma containing -tocopherol reacts with ferric chloride, thus reducing the ferric ions to ferrous ions. The ferrous ions then react with, ’ – dipyridyl to give a red coloured complex which is measured at 520 nm. Carotenoids, which were also extracted into xylene, were estimated by their absorbance at 460 nm and a correction was applied at 520 nm. The carotenoid absorption at 520 nm was 29% of the absorption at 460 nm.

Retinol (9)
Proteins get precipitated on the addition of ethanol and the concentration of Retinol can be determined by reading the extinction value of the heptane extract of retinol at 327nm.

Ascorbic Acid (10)
When ascorbic acid reacts with 2, 6-dichlorophenol indophenol, reduced 2, 6-dichlorophenol indophenol is formed which is colourless. The decrease in colour is proportional to the concentration of ascorbic acid which is present in the solution. Decrease in the optical density was measured at 520nm and the concentration was calculated from standards which were treated similarly.

Statistical analysis of all the obtained parameters in patients with chronic renal failure (before and after dialysis) and the control groups were done using paired and unpaired student’s ‘t’ test. Also, Logistic regression analysis was performed for all the parameters.

Results

(Table/Fig 1) shows the mean and standard deviation of oxidant and anti-oxidant levels in controls and in CRF patients, before and after dialysis. The levels of MDA were significantly increased in pre-dialysis CRF patients as compared to the controls (p<0.001) and the levels were further significantly increased in post-dialysis patients as compared to the pre-dialysis patients (p<0.001). The levels of Vitamin E, Vitamin A and Vitamin C showed significant decrease in pre-dialysis CRF patients as compared to healthy controls (p<0.001) and further significant decrease was seen in post-dialysis CRF patients as compared to the pre-dialysis patients (p<0.001).The following variables (MDA, Vitamin E, Vitamin A and Vitamin C) were also considered for multiple regression analysis as shown in (Table/Fig 2) , (Table/Fig 3). The model used for regression analysis showed that the R square was 0.75. By this, we can explain that 75% of the changes in the dependent variable (MDA) can be attributed to the variables which were considered for the regression (Vitamin E, Vitamin A and Vitamin C).

Discussion

CRF leads to imbalance between the production of oxygen free radicals and cellular antioxidants system. The Hexose Mono Phosphate (HMP) shunt pathway which is required for NADPH synthesis in RBC is impaired in CRF, thus resulting in the accumulation of free radicals. This enhances the process of oxidative stress and plays a role in the development of the disease, thus leading to an increase in MDA levels (11), (12),(13). Further increase in the levels of MDA in post-dialysis CRF patients may be due to the exacerbation of the lipid peroxidation process by haemodialysis. Haemodialysis leads to the activation of complement system, which in turn stimulates free radical generation, mainly superoxides through the NADPH system. Heparin which is used in haemodialysis causes lipolysis, thus increasing free fatty acid in plasma, which leads to the exacerbation of lipid peroxidation. The red cell MDA increases the rigidity of the RBCs and makes them more susceptible to lysis during haemodialysis (14), (15).

Antioxidants
In the present study, plasma concentrations of Vitamin E, Vitamin A and Vitamin C were significantly decreased. The levels were further significantly decreased in post-dialysis CRF patients as compared to pre-dialysis CRF patients.

Plasma -tocopherol (Vitamin E)
Decreased levels of -tocopherol in predialysis CRF patients as compared to healthy controls may be due to enhanced lipid peroxidation. There may also be impaired absorption of dietary -tocopherol due to altered lipid metabolism. After dialysis, further decreased levels may be due to increased consumption in an attempt to reduce the effect of the oxygen free radicals (16), (17), (18).

Plasma Vitamin A
Decreased levels of Vitamin A in predialysis CRF patients as compared to healthy controls, was due to enhanced lipid peroxidation, thus leading to the exhaustion of hepatic storage. So, there was reduced conversion of -carotene to vitamin A and decreased secretion of vitamin A from the liver into the circulation. In post-dialysis CRF patients, the enhanced production of oxygen free radical leads to the increased utilization of vitamin A and its microsomal degradation, thus resulting in its reduced levels (14).

Plasma Vitamin C
Decreased levels of Vitamin C in pre-dialysis CRF patients as compared to healthy controls, may be due to the utilization of ascorbic acid to generate -tocopherol from the -tocopheroxyl radical at the water lipid interface. The ascorbate may be reduced because it is an efficient quencher of superoxide peroxyl and hydroxyl radicals. In addition to the above, there may be associated nutritional deficiency of ascorbate to limit potassium intake in CRF patients. During haemodialysis, there may be additional loss of ascorbic acid from plasma, as it is a water soluble vitamin (14),(16),(17).

Conclusion

The result of the present study indicates that there is considerable oxidative stress in patients with CRF, which is further exacerbated by haemodialysis, as evidenced by increased lipid peroxidation and low antioxidant levels. Increased levels of Malondialdehyde (MDA) which is a reliable marker and a product of lipid peroxidation in CRF and dialysis patients, indicates the existence of oxidative stress.

The decreased levels of non-enzymatic antioxidants like Vitamin E, Vitamin A and Vitamin C indicate the increase in oxidative stress. Inflammatory processes during dialysis appear to be the main factors which are involved in oxidative stress. Exogenous supplementation of non-enzymatic antioxidants may decrease the damage to renal tissue by quenching and preventing the free radical action which are responsible for the disease process.

References

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M Taccone Gallucci, R Lubrano, C Meloni. Vitamin E as antioxidant agent; in Ranco C, La Greca G; Vit E bonded membrane. A further step in dialysis optimization. Contrib. Nephrol Basel, Karger 1999; 127:32-43.
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