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

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Dr Bhanu K Bhakhri

"The Journal of Clinical and Diagnostic Research (JCDR) has been in operation since almost a decade. It has contributed a huge number of peer reviewed articles, across a spectrum of medical disciplines, to the medical literature.
Its wide based indexing and open access publications attracts many authors as well as readers
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Dr Bhanu K Bhakhri
Faculty, Pediatric Medicine
Super Speciality Paediatric Hospital and Post Graduate Teaching Institute, Noida
On Sep 2018




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Believers Church Medical College,
Thiruvalla, Kerala
On Sep 2018




Prof. Somashekhar Nimbalkar

"Over the last few years, we have published our research regularly in Journal of Clinical and Diagnostic Research. Having published in more than 20 high impact journals over the last five years including several high impact ones and reviewing articles for even more journals across my fields of interest, we value our published work in JCDR for their high standards in publishing scientific articles. The ease of submission, the rapid reviews in under a month, the high quality of their reviewers and keen attention to the final process of proofs and publication, ensure that there are no mistakes in the final article. We have been asked clarifications on several occasions and have been happy to provide them and it exemplifies the commitment to quality of the team at JCDR."



Prof. Somashekhar Nimbalkar
Head, Department of Pediatrics, Pramukhswami Medical College, Karamsad
Chairman, Research Group, Charutar Arogya Mandal, Karamsad
National Joint Coordinator - Advanced IAP NNF NRP Program
Ex-Member, Governing Body, National Neonatology Forum, New Delhi
Ex-President - National Neonatology Forum Gujarat State Chapter
Department of Pediatrics, Pramukhswami Medical College, Karamsad, Anand, Gujarat.
On Sep 2018




Dr. Kalyani R

"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



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Professor and Head
Department of Pathology
Sri Devaraj Urs Medical College
Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
On Sep 2018




Dr. Saumya Navit

"As a peer-reviewed journal, the Journal of Clinical and Diagnostic Research provides an opportunity to researchers, scientists and budding professionals to explore the developments in the field of medicine and dentistry and their varied specialities, thus extending our view on biological diversities of living species in relation to medicine.
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Dr Saumya Navit
Professor and Head
Department of Pediatric Dentistry
Saraswati Dental College
Lucknow
On Sep 2018




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MD, DM (Clinical Pharmacology)
Assistant Professor
Department of Pharmacology
Calcutta National Medical College & Hospital , Kolkata




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Best regards,
C.S. Ramesh Babu,
Associate Professor of Anatomy,
Muzaffarnagar Medical College,
Muzaffarnagar.
On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



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




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
The journal has a monthly publication and the articles are published quite fast. In time compared to other journals. The on-line first publication is also a great advantage and facility to review one's own articles before going to print. The response to any query and permission if required, is quite fast; this is quite commendable. I have a very good experience about seeking quick permission for quoting a photograph (Fig.) from a JCDR article for my chapter authored in an E book. I never thought it would be so easy. No hassles.
Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help ones reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journalsNo manuscriptsNo authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



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.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
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

Important Notice

Original article / research
Year : 2010 | Month : February | Volume : 4 | Issue : 1 | Page : 2061 - 2067

A Potential Correlation between Systemic Oxidative Stress and Intracellular Ambiance of the Lens Epithelia in Patients with Cataract

GOYAL M M *, VISHWAJEET P*, MITTAL R** , SUNE P***

Dept. of Biochemistry*, Dept. of Anatomy**,Dept.ofOphthalmology***,J.N.Medical College,Datta Meghe Institute of Medical Sciences University,Wardha (MH), (INDIA).

Correspondence Address :
Madhur M. Goyal,Assistant Professor
Department of Biochemistry,J.N.Medical]
College,Datta Meghe Institute of
Medical Sciences
University, Wardha-442001 (MH),(INDIA),
Ph:919823585404,E.mail :monusvm@yahoo.com

Abstract

Aim: To study the correlation between systemic oxidative stress and intracellular ambiance of the lens epithelia in patients with cataract.
Materials and Methods: Spectrophotometery was employed for the estimation of catalase activity and the extent of lipid peroxidation in the lens epithelial cells (LEC) and plasma. Both are markers for oxidative stress. No antioxidant medicines were used by the cataract patients enrolled in this study; otherwise, they were all healthy individuals without any systemic diseases.
Results: A total of 56 patients with cataract were included in this study. The mean ages of the patients were 66.6 ± 8.3 (±SD) years for males and 62.4 ± 10.0 years for females. Catalase activity was estimated in surgically removed LEC (221.16±135.87 U/µg protein; Mean±SD) and in plasma (277.56±162.44 KU/mg Hb). MDA levels were also calculated in LEC (1.28±0.79nM/mg protein) and in plasma (537.30±238.47 nM/g protein). Linear regression analysis showed a partial positive correlation in LEC and plasma catalase activity (r: 0.701; p<0.05), but not in MDA (r: 0.248; p>0.05).
Conclusion: Increased systemic oxidative stress can lead to the development or progression of cataract by affecting the intracellular ambiance of the lens epithelia. So, subjects having high systemic oxidative stress are more vulnerable for the development of cataract.

Keywords

cataract formation, lens epithelium, catalase, lipid peroxidation

Introduction
Cataract is the leading cause of blindness worldwide. In India, contemporary findings suggest that the population is either widely exposed to the environmental risk factors and /or there is a genetic predisposition. In a recent study on the North Indian rural population, the prevalence of cataract in the age group >50 years was 75.3% (1), which appears to be the highest in the world. Even with all resources available, clinicians are finding it hard to eradicate cataract induced blindness completely (2). Therefore, it is indispensable to find out the causative factors for the prevention of the development of cataract.

Several risk factors have been identified for the development of human cataract: aging, diabetes, malnutrition, poverty, sunlight, smoking, hypertension, and renal failure (3). Although cataract is a multifactorial disease, oxidative mechanisms are believed to play an important role in the pathogenesis of maturity-onset cataract, the most important cause of visual impairment at an advanced age. Oxidative stress (OS) occurs when the level of pro-oxidants (reactive oxygen species and other free radicals) exceeds the ability of the cell to respond through antioxidant defenses and ultimately leads to the modification and degradation of protein, damage to DNA and mitochondria and cell death (4). The increased production of free radicals and the oxidation of unsaturated lipids have been observed in cataractous lenses (5) and aqueous humour. Lens epithelial cells (LEC) are the main site of oxidative damage, which then ultimately affects the entire lens, thus affecting the lens clarity and leading to the pathogenesis of cataract (6), (7).

On the other hand, since many years, experimental evidences have suggested an association between nutrition and lens opacities. Increased systemic OS has been observed in cataract patients (8]. In some other studies, the parameters of their antioxidative defenses are lower and those of OS are higher in serum, in the lens and in the humour aqueous (9).

But systemic stress (oxidative) and its effects on the intracellular ambiance of the lens epithelia have not been correlated directly in pathological human samples. Therefore, the present study was undertaken to examine the correlation between oxidative stress in surgically removed LEC and plasma of patients with cataract.

Material and Methods

Patient Selection
This study comprised of 56 consecutive patients with uncomplicated age-related cataract, who had undergone phacoemulsification. Informed consent was obtained from all patients. Only patients older than 50 years, with pupils dilating more than 7.0 mm and with otherwise normal eyes, were included in the study. The exclusion criteria were diabetes mellitus, hypertension, glaucoma, shallow anterior chamber, high myopia (axial length O27.0 mm), pseudoexfoliation, traumatic cataract, subluxated cataract, previous ocular surgery, ocular disease, steroid or immunosuppressive therapy and allergy to dilating drops.

Collection and Processing Of Samples
Circular pieces of cataractous human anterior lens capsule (rhexis), about 6mm in diameter, were obtained post operatively in sterile normal saline. The cataract type was noted using the LOCs classification system (10). The anterior capsules taken for all the experiments belonged to immature senile cataract. For acclimatization, a single rhexis was placed in 1 ml of Eagle’s Minimal Essential Medium (MEM) containing 10% foetal calf serum in a single well of a 24 well plate and was incubated for 30 min in a 5% CO2 incubator at 370C.

Fasting blood samples were collected in a glass bulb containing EDTA from patients, and their verbal consent was taken. The plasma was separated and stored at -20 °C until analysis.

Cell Viability Test
The Typan blue exclusion test was used to ensure cell viability. In this test, a few drops of typan blue is added on a rhexis, placed on a glass slide and then microscopically examined to determine whether cells take up or exclude the dye. A viable cell will have a clear cytoplasm whereas a nonviable cell will have a blue cytoplasm.

Biochemical Assay
The level of malondialdehyde (MDA) was measured as an index of peroxidation of lipids by using the method of Beuge and Aust (11). The catalase activities of plasma and LECs were measured by Luck’s method (12). The catalase specific activity was calculated in LEC by measuring protein using the Eosin Y method (13) and in plasma by estimating haemoglobin using the O-tolidine method (14). The methods are described here in brief.

Catalase Assay
The enzyme extract was prepared by homogenizing the cells in a lysis buffer (0.25M sucrose, 20mM tris-HCl,100mM KCl, 40mM NaCl, and 10mM MgCl2), centrifuging them at 20 000 g for 30 min at 41C and by re-extracting the pellet in a microsomal dilution buffer (0.1M KH2PO4, 20% glycerol, 10mM EDTA, and 0.1mM b-mercaptoethanol). The pooled extract was added to 10mM of H2O2 and the rate of decomposition of H2O2 was measured spectophotometrically at 240 nm. The decomposition of 1 mM of H2O2 in 1 min corresponds to 1U of catalase.

Lipid peroxidation / MDA assay
TBA-TCA- HCl reagent (0.8 ml) was added to 0.2 ml of plasma. The mixture was boiled in a boiling water-bath for 15 min. After centrifugation, the absorbance of the supernatant was recorded spectrophotometrically at 535 nm. A blank (saline) absorbance due to reagents was subtracted from the corresponding experimental sample. The MDA content in the sample was calculated using an extinction coefficient of 1.56 × 105 /M/cm. at 535 nm. In LEC, the cells were homogenized in distilled water and were centrifuged at 12,000 g at 4°C for 30 min. Aliquots of this homogenate were used as the sample instead of plasma in the above procedure.

Estimation of Protein
100µl of the sample was added to 1.0 ml of the reagent (0.012% Eosin Y Dye and 0.6% Citric Acid; pH: 2.6-2.8) and was incubated for 15min at room temperature. The absorbance was measured spectrophotometrically at 543nm.

Estimation of Plasma Haemoglobin
1.0 ml of working solution [0.4 gm % o-tolidine, 0.1% triton x-100 in solution (20:80) of glacial acetic acid and ethanol] and 1 ml of freshly prepared H2O2 solution [2% H2O2 and 2.26 % w/v sodium acetate in distilled water] were mixed in test tubes and this was kept for 5 min for maturation of the reagent. 10 µL of haemoglobin standards (6 to 400 mg/L) and samples were added into respective test tubes and immediately, ∆Abs per minute were measured at 630 nm with a delay time of 30 secs.Distilled water was used as the reagent blank.


Results

A total of 56 rhexis samples were collected from patients with immature senile cataract, having mean ages of 66.6 ± 8.3 (±SD) years for males and 62.4 ± 10.0 years for females. They (males and females) were approximately equal in number. We analyzed 25 samples for MDA and 31 samples for catalase activity separately, in LEC and plasma of the same patients.

Cell Viability and Acclimatization
It was observed that the lens epithelial cells which were present in the capsulorhexis remained viable upto 2 hours in PBS, as shown in figure 1A. More than 90 % of the cells had clear cytoplasm (live cells) up to 2 hours, whereas the remaining cells picked up the dye (dead cells). LECs rapidly died after 2 hours in PBS (Figure 1B) (Table/Fig 1). However, in the present study, it took a maximum of twenty minutes to enter the sample into the process after its removal from the eye.

Correlation between Catalase Activity in Plasma and in LEC
Catalase specific activity was measured both in LEC (mean ± SD; 221.16 ± 135.87 U/µg of protein) as well as in plasma (277.56 ± 162.44 KU/mg of Hb) of the same patient. A partial positive linear correlation was found (r: 0.701, p<0.05) between the catalase activity in LECs and in the plasma of the patients with cataract (Table/Fig 2).

Correlation between MDA in Plasma and in LEC
MDA level was measured in LECs (1.282 ± 0.798 nM/mg protein; mean± SD) and in plasma (537.30 ± 238.47 nM/g protein) of the same patient. As shown in (Table/Fig 3), MDA levels in LECs are poorly correlated (positive) with plasma MDA levels. The correlation coefficient (r) was only 0.248. It was in accord with the results of other workers (15).

Discussion

The elevated level of oxidative stress markers are reported in serum and RBCs in cataract patients. Experimental and observational data suggest that oral supplementation or increased blood level of micronutrients which can reduce oxidative stress, can retard the development of age-related cataract (16),(17),(18),(19).

In the present study, we attempted to find out the correlation between systemic and LEC oxidative stress. In the previous studies whole lens cells (5),(8) or aqueous humour samples were used to see the correlation of different factors in ocular tissue and blood. The generation of free radicals is an unwanted surplus of metabolism and it is a continuous process in living cells. Most of the lens cells (except epithelial cells) and aqueous humour do not contain cell organelles and so they are not metabolically active. Lens contents are a result of diffusion from LECs. So, appraisal of the effects of systemic oxidative stress on the lens leave a question as to how these cells which are not capable of general metabolism, react against stress.

Again, there are other factors also which can induce free radical generation in the surroundings of the lens, like UV light. The aqueous humour normally contains hydrogen peroxide (H2O2), a compound which is capable of generating reactive oxygen species (ROS). The systems protecting the ocular lens from these ROS (or oxidative damage) are primarily confined to the epithelium, a single layer of cells on the anterior side of the organ which directly lies beneath the lens capsule (20). These LECs are centres of lens metabolic activities as they contain maximum mitochondria than any other parts of the lens (21). Some data suggest that these cells are the initial site of attack by oxidative stress and follows the lens fibers, leading to cortical cataract (22),(23). Hence, LECs have great significance in cataractogenesis and were employed to study the changes which were brought about by oxidative stress (22). Cataract surgery can induce some stress in LEC (24). To avoid this, the acclimatization step was introduced. The same model was previously used by our co authors also (25),(26)[,27].

Since ROS are highly unstable, their measurement in human serum or plasma samples does not necessarily reflect their in vivo concentration. Therefore, only secondary (end) products of oxidative stress have emerged for application in clinical studies. Malondialdehyde (MDA) is a naturally occurring product of lipid peroxidation, a well-established mechanism of cellular injury by ROS in both plants and animals and is used as an indicator of oxidative stress in cells and tissues.

H2O2 is the major oxidant involved in cataract formation (28). There is a significant increase in the H2O2 concentration during cataractogenesis (17). In in-vitro conditions also, relatively high concentrations of H2O2 are needed to cause significant changes in the lens epithelial cells (4). Two major antioxidative enzymes used by the lens to combat H2O2 are catalase and glutathione peroxidase (29). Glutathione peroxidase plays a major role in removing H2O2 which is present in low concentrations in cells, whereas catalase is more effective with high concentrations of H2O2 (30). In another report, the cause of cell death following the inhibition of catalase was identified to be related to an inability of the cultured LEC to remove peroxide from the culture medium at a rapid rate, following the H2O2-pulse (31). Catalase also protects other antioxidant enzymes from the destructive effects of H2O2 (32). Hence, catalase was preferred to glutathione peroxidase and was used as an indirect marker to evaluate the oxidative status of the sample. In the lens of an experimental model, a slight stimulation of the antioxidant systems by a small number of free radicals was observed, which provoked a reaction of sweeping them away (33). Perhaps by the same mechanism, its (catalase) activity is increased to counteract increased oxidative stress (in term of H2O2) either in LEC or in blood (systemic) (34).

Both catalase activity and MDA levels are good markers of oxidative stress. An increase in plasma catalase activity and MDA reflect increased systemic oxidative stress which might be the result of poor nutrition or smoking and tobacco chewing habits or environmental exposure, or all of them. Body tissue including LEC can react against oxidative stress in two ways. One is, by increasing the synthesis of antioxidant enzymes like catalase and glutathione peroxidase and the second is through more consumption of antioxidant substrates like reduced glutathione, vitamin C and vitamin E. Due to increased OS, the consumption of antioxidant substrates by body tissues increases and therefore, its availability is reduced across the blood brain barrier (BBB), a membranic structure that acts primarily to protect the brain from chemicals in the blood, while still allowing essential metabolic function. In this way, slight decrease in the availability of antioxidant substrates to LEC, which is continuously exposed to factors which provoke free radical generation like light, leads these cells to OS. If this condition persists for a long time, it affects the whole lens and its clarity by modification or degeneration of DNA, proteins and other biomolecules like lipids. These cells also respond by increasing the synthesis of catalase and if fails to respond, cataract develops. This is indicated by some findings where the catalase activity in cataract patients was significantly lower than in the control subjects (35). Linear regression analysis results (Figure 2) endorsed the above said matter and found a correlation between the catalase activity in LEC and in the plasma of same patient (r : 0.701, p<0.05). These results are in agreement with those of other groups who worked on the same hypothesis and reported a correlation between different enzymes and lipidperoxidation products (36). Some contradictory reports which are also available (6),(15), might be the result of the presence of factors other than elevated levels of H2O2. We could not find any report in which the catalase activities in surgically removed LEC and in plasma were correlated.

Plasma MDA levels are poorly correlated (Figure 3) with MDA levels in LECs. In the previous studies, an inverse correlation of MDA and catalase activity has been reported. It is quite obvious as both are produced by opposite mechanisms. Lipid modification (peroxidation) by ROS results in an increase in MDA levels whereas increased catalase activity shows an elevated defense mechanism in the cells. This might be the reason for the poor correlation (Figure 3), as increased catalase activity decreases oxidative stress in LEC and inhibits lipid peroxidation (formation of MDA).

Although our results are from a relatively small sample size, they suggest some intriguing and potentially provocative findings which suggest that systemic oxidative stress can increase the same (OS) in LECs. This change in the intracellular ambiance of the lens epithelia can lead to modification of DNA, proteins and other biomolecules and can result in the development or progression of lens opacity. From this, we can conclude that individuals having increased systemic oxidative stress are more vulnerable to the development of cataract and so the oral supplementation or increased blood levels of the antioxidants may be beneficial in the prevention of cataractogenesis. Moreover, further studies with an increased sample size and different stress markers are needed to define the unambiguous role of oxidative stress in the development of cataract.

Acknowledgement

We are thankful to Dr Vijay Babar, Assist. Professor,Department of Medical Education, DMIMS, Wardha, for his help as a statistician.

References

1.
Murthy GV, Gupta SK, Maraini G, Camparini M, Price GM, Dherani M, John N, Chakravarthy U, Fletcher AE. Prevalence of lens opacities in North India: the INDEYE feasibility study. Invest Ophthalmol Vis Sci. 2007; 48(1):88-95.
2.
Murthy GVS, Gupta SK, John N, Vashist P. Current status of cataract blindness and Vision 2020: The right to sight initiative in India. Indian J Ophthalmol: 2008; 56:489-94.
3.
Harding J. The epidemiology of cataract. In: Harding J, ed. Cataract - biochemistry, epidemiology and pharmacology. 1st ed. Madras Chapman and Hall, 1991: 83.
4.
Doshna CM, Shen AC, Gebhard DF, Brees DJ, Somps CJ. Investigating markers of DNA oxidation, lipid peroxidation and stress response in human lens epithelial cell lines. Invest Ophthalmol Vis Sci 2005; 46: E-Abstract 3850.
5.
Ferrari MT, Vendemiale G, Grattagliano I, Boscia F, Arnese L, Altomare E, Cardia L. Role of lipid peroxidation in the pathogenesis of myopic and senile cataract. British Journal of Ophthalmology 1996; 80:840-843.
6.
AREDS Report No. 9. A randomized, placebo-controlled, clinical trial of high-dose supplementation with vitamins C and E and beta carotene for age-related cataract and vision loss: Arch Ophthalmol 2001; 119(10): 1439–1452.
7.
Fan X, Reneker LW, Obrenovich ME, Strauch C, Cheng R, Jarvis SM, Ortwerth BJ, Monnier VM. Vitamin C mediates chemical aging of lens crystallins by the Maillard reaction in a humanized mouse model. PNAS 2006; 103 (11): 16912–16917.
8.
Donma O, Yorulmaz E, Pekel H, Suyugul N. Blood and lens lipid peroxidation and antioxidant status in normal individuals, senile and diabetic cataractous patients. Curr Eye Res 2002; 25(1):9-16.
9.
Zorić L. Parameters of oxidative stress in the lens, aqueous humor and blood in patients with diabetes and senile cataracts. Srp Arh Celok Lek 2003; 131(3-4):137-42.
10.
Chylack LT Jr, Wolfe JK, Singer DM, Leske MC, Bullimore MA, Bailey IL, Friend J, McCarthy D, Wu SY. The Lens Opacities Classification System III. The Longitudinal Study of Cataract Study Group. Arch Ophthalmol 1993; 111(6):831-6.
11.
Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol 1978; 52: 302–310.
12.
Luck H. Catalase. In: Bergmeyer HU, editor. Methods of enzyme analysis. Academic Press: New York; 1971. p. 885.
13.
Waheed AA, Gupta PD. Estimation of submicrogram quantities of protein using the dye eosin Y. J Biochem Biophy methods 2000; 42:125-132.
14.
Goyal MM, Basak A. Estimation of plasma haemoglobin by a modified kinetic method using o-tolidine. Indian Journal of Clinical Biochemistry 2009; 24 (1): 36-41.
15.
Lian H, Li S, Cao X, Pan S, Liang S. Malonaldehyde, superoxide dismutase and human cataract. Yan Ke Xue Bao 1993; 9(4):186-9.
16.
Jacques PF, Hartz SC, Chylack LT, McGandy RB, Sadowski FA. Nutritional status in persons with and without senile cataract:blood vitamin and mineral levels. Am J Clin Nutr l988; 48:l52-8.
17.
Bhuyan KC,Bhuyan DK. Molecular mechanism of cataractogenesis III.Toxic metabolites of oxygen as initiators of lipid peroxidation and cataract. Curr Eye Res 1984; 3(1):67-81.
18.
Jacques PF. Nutritional antioxidants and prevention of age related eye disease. Antioxidant and disease prevention. E.D. Garewal H.S. CRC Press NY; 1997: p. 179-173.
19.
Harding J.Cataract - Biochemistry, Epidemiology and Pharmacology. St. Edmand sbury press N.Y.; 1991: 83.
20.
Reddan JR, Steiger CA, Dziedzic DC, Gordon SR. Regional differences in the distribution of catalase in the epithelium of the ocular lens. Cell Mol Biol 1996; 42(2):209-19.
21.
McNulty R,Wang H,Mathias RT, Ortwerth BJ, Truscott RJW, Bassnett S. Regulation of tissue oxygen levels in the mammalian lens. J Physiol 2004; 559 (3): 883-898.
22.
Kalariya N, Rawal VM, Vasavada AR. Human lens epithelial layer in cortical cataract. Indian J Ophthalmol 1998; 46:159-62.
23.
Spector, A. Oxidative stress induced cataract: Mechanism of action. FASEBJ 1995; 9: 1173-1182.
24.
Augustin AJ, Dick HB.Oxidative tissue damage after phacoemulsification: influence of ophthalmic viscosurgical devices. J Cataract Refract Surg 2004; 30: 424-7.
25.
Rajkumar S, Praveen MR, Gajjar D, Vasavada AR, Alapure B, Patel D, Kapur S. Activity of superoxide dismutase isoenzymes in lens epithelial cells derived from different types of age-related cataract. J Cataract Refract Surg 2008;34(3):470-4.
26.
Gajjar D, Patel D, Alapure B, Praveen MR, Patel A, Johar K Sr, Vasavada AR. Rapid action of oestradiol against hydrogen peroxide-induced oxidative stress in cataractous lens epithelium: an in vitro study. Eye. 2008; 1-8.
27.
Goyal MM, Gajjar DU, Patel DB, Sune P, Vasavda AR. Effect of vitamin C and E activity on surgically removed cataractous human lens epithelium cells. IJCB 2009; 24(4): 375-380.
28.
Spector A. Oxidative stress-induced cataract: mechanism of action. FASEB J 1995; 9: 1173-1182.
29.
Spector A, Ma W, Wang RR, Yang Y, Ho YS. The contribution of GSH peroxidase-I, catalase and GSH to the degradation of H2O2 by the mouse lens. Exp Eye Res 1997; 64:477-485.
30.
Costarides AP, Riley MV, Green K. Roles of catalase and the glutathione redox cycle in the regulation of anterior-chamber hydrogen peroxide. Ophthalmic Res 1991; 23(5):284-94.
31.
Giblin FJ, Reddan JR, Schrimscher L, Dziedzic DC, Reddy VN. The relative roles of the glutathione redox cycle and catalase in the detoxification of H2O2 by cultured rabbit lens epithelial cells. Exp Eye Res 1990; 50(6):795-804.
32.
Bhuyan KC,Bhuyan DK. Suproxide dismutase of the eye: relative functions of superoxide dismutase and catalase in protecting the ocular lens from oxidative damage. Biochim Biophys Acta1978; 542(1):28-38.
33.
Sztarbała T, Goś R, Kedziora J, Błaszczyk J, Sibińska E, Góralczyk M. Changes in the antioxidant system of the aqueous humor, lens and erythrocytes after sulfur hexafluoride. Klin Oczna 1998; 100: 73-5.
34.
Chandrasena LG, Chackrewarthy S, Teckla P, Perera MJ, Silva D. Erythrocyte antioxidant enzymes in patients with cataract. Annals of Clinical & Laboratory Science 2006; 36:201-204.
35.
Ates NA, Yildirim O, Tamer L, Unlu A, Ercan B, Muslu N, Kanik A, Hatungil R, Atik U. Plasma catalase activity and malondialdehyde level in patients with cataract. Eye 2004; 18(8): 785-788.
36.
Rola JK, Zagórski Z. Peroxidation of lipids in patients with senile cataract. Klin Oczna 2004; 106: 416-8.

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