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

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On Sep 2018




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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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Sri Devaraj Urs Academy of Higher Education and Research , Kolar, Karnataka
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Saraswati Dental College
Lucknow
On Sep 2018




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Muzaffarnagar.
On Aug 2018




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"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.
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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 one’s 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.


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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.
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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
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 : 2022 | Month : March | Volume : 16 | Issue : 3 | Page : AC10 - AC14 Full Version

Neurobehavioural and Neurochemical Changes in Arsenic Induced Cerebellar Toxicity in Male Sprague-Dawley Rats: An Experimental Study


Published: March 1, 2022 | DOI: https://doi.org/10.7860/JCDR/2022/53048.16157
Ravi Shankar Prasad Sawan, Sridevi Nangali Srinivasa, Shashidhar Kurpad Nagaraj

1. PhD Scholar, Department of Anatomy, Sri Devaraj Urs Medical College, Kolar, Karnataka, India. 2. Professor and Head, Department of Anatomy, Sri Devaraj Urs Medical College, Kolar, Karnataka, India. 3. Professor and Head, Department of Biochemistry, Sri Devaraj Urs Medical College, Kolar, Karnataka, India.

Correspondence Address :
Sridevi Nangali Srinivasa,
Professor and Head, Department of Anatomy, Sri Devaraj Urs Medical College, Kolar, Karnataka, India.
E-mail: sridevins26@gmail.com

Abstract

Introduction: Sodium arsenite, an inorganic arsenic, is naturally present at high level (>50 μg/L) in ground water. Drinking ground water is the biggest threat to public health. Though, there are numerous reports on arsenic neurotoxicity, the arsenic effect on cerebellar neurotoxicity remains vague especially its chronic effect on its neurobehavioural and neurochemical alterations.

Aim: To evaluate the neurobehavioural and neurochemical alterations caused by sodium arsenite in cerebellum of rats.

Materials and Methods: This experimental study was conducted in the Central Animal House at Sri Devaraj Urs Academy of Higher Education and Research (SDUAHER) from November 2019 to February 2020 for a period of 90 days. Total 16 male sprague-dawley rats were randomised into two equal groups. Group I: Control, received normal saline. Group II: Sodium arsenite, doses of 50 Parts per Millions (PPM) for 90 days through oral gavage. Rats were subjected to Open Field Test (OFT) for locomotor and exploratory behaviour and Beam Walking Test (BWT) for motor coordination and balance. Following behavioural tests, rats were anaesthetised. Blood was drawn from a retro-orbital puncture. Brains were dissected and cerebellum was separated. Concentration of Malondialdehyde (MDA), Nitric Oxide (NO) and activity of Glutathione Peroxidase (GPx) were assessed spectrophotometrically in serum and cerebellum of rats. Mean±SD was used for normally distributed data and groups were compared using independent t-test, whereas for non-normally distributed data, Median (25th-75th Percentile) was used and Mann-Whitney U-test was used to compare groups.

Results: Arsenic-treated rats showed a significant increase in arsenic concentration in serum and cerebellum (5.5±1.6 ng/mL, 2.76±0.56 μg/g, respectively) compared to control (1.14±0.43 ng/mL, 0.65±0.29 μg/g, respectively). There was a significant decrease in locomotor and exploratory behaviour and impairment in motor coordination and balance in arsenic treated rats with a p-value <0.001 in comparison with control rats. The arsenic treated rats had significantly enhanced concentration of MDA and NO level and reduced activity of GPx in serum {16.84 (13.84-18.87), 33.79 (30.05-37.17) nmol/mL, and 6.89 (5.24-8.5) mmoles of Reduced glutathione (GSH) oxidised/min/mL, respectively} compared to control {8.81 (8.36-9.48), 17.66 (15.33-21.29) nmol/mL, and 15.16 (12.77-16.59) mmoles of GSH oxidised/min/mL, respectively} and also found increased concentration of MDA and NO level and reduced activity of GPx in tissue {7.98 (7.14-8.92), 24.67 (21.4-28-22) nmol/mg of protein and 2.66 (1.19-3.86) mmoles of GSH oxidised/min/mg protein, respectively} compared to control {3.02 (2.35-3.61), 13.93 (11.0-16.16) nmol/mg of protein and 7.63 (7.08-9.19) mmoles of GSH oxidised/min/mg protein, respectively}.

Conclusion: The oral administration of sodium arsenite at the doses of 50 PPM for 90 days showed interesting alterations in neurobehavioural and neurochemical parameters related to cerebellum of rats.

Keywords

Cerebellum, Glutathione peroxidase, Motor activities, Oxidative stress markers

In 21st century world, industrialisation of society and expansion of factories has vividly expanded the amount of pollutants and environmental pollutions. Environmental pollution due to arsenic is a major toxin and has become a major public health challenge in many countries such as Bangladesh, India, Nepal, Taiwan, Mongolia, Vietnam, Pakistan, China, Afghanistan, Argentina and USA (1). Arsenic is mainly distributed in the environment through industries using wood preservatives, glass, semiconductors, mining waste, herbicides and pesticides (2). Due to its increasing production and utilisation, arsenic is ubiquitously present in water, air and soil affecting occupational workers as well as general population (3).

Arsenic primarily enters into the body through direct consumption of drinking water from geological deposits by drilling of tube well (4). World Health Organization (WHO) and other regulatory bodies have established a Maximum Contaminant Limit (MCL) of 10 μg/L inorganic arsenic (iAs) in drinking water for the safety of human health based on reducing cancer risk, but this limit is not considering endpoints other cancer end points such as digestive effects, reproductive effect (5). Globally, more than 200 million people are affected by ground-water contaminated with arsenic concentration greater than 10 μg/L (6). Arsenic affected area in West Bengal and Bihar region of the Ganga Plain in India has extremely high arsenic contaminated water where arsenic concentration in water has been reported upto 1000 μg/L (i.e.,100 PPM) against the safe limit of 10 μg/L recommended by WHO (7),(8). High dosages iAs can cause death but chronic lower levels of arsenic results in serious health problems such as cancers and skin lesions (9). Numerous experimental and epidemiological data provide evidence that acute and chronic arsenic exposure has been linked with numerous chronic indices in all human and animal organ systems and contributes to wide spectrum of diseases, especially diseases involving central nervous system (10),(11),(12).

Cerebellum is an important component of brain which is linked to control of posture, coordination of movements, gait and skilled voluntary movements. Some studies suggest that cerebellum is also involved in cognition, language and executive functioning (13). Arsenic penetrates the Blood Brain Barrier (BBB) and accumulates in different regions of the brain such as cortex, cerebellum and hippocampus and increases the production of reactive oxygen species in brain which causes an imbalance between the anti-oxidant defense system levels and the production of free radicals to prompt neurotoxicity (14),(15).

Arsenic induced oxidative stress causes neuronal injuries leading to neuronal cell death and impairment of brain functions because of its high energy demand, high oxygen consumption and abundance of polyunsaturated lipid contents and have shown various neurological side-effects such as vertigo, impaired coordination of movements, uncontrolled motor learning, unsteady gait, loss of skilled voluntary movements, impaired learning, memory and concentration (16),(17),(18). Reports in the literature also documented that arsenic-treated rats reduces neuronal viability in primary cultures of rat cerebellar neurons and showed alterations in locomotor behaviour and learning task (19),(20).

These literatures indicate that arsenic has an impact on the cerebellum. There are minimal studies with a possible impact of arsenic on neurobehavioural and biochemical changes related with cerebellum (21),(22). Considering the lacunae, the present study aimed to investigate the impact of arsenic in cerebellum associated neurotoxicity by evaluating motor coordination, balance, locomotor and exploratory activities of rats with standard neurobehavioural procedure and also evaluated the biomarkers of oxidative stress.

Material and Methods

This experimental study was conducted in the Central Animal House at Sri Devaraj Urs Academy of Higher Education and Research (SDUAHER) from November 2019 to February 2020 for a period of 90 days. Full details of work plan with experimental animals were approved by Institutional Animal Ethics Committee (IAEC) (IAEC/PHARMA/SDUMC/2018-19/12a from SDUAHER, Kolar). All experiments were made according to guidelines established by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) (23). All efforts were made to minimise animal suffering and to reduce the number of animals used.

Inclusion criteria: Healthy male Sprague-Dawley rats having 150 to 180 grams in weight were included in the study.

Exclusion criteria: Rats with any signs/symptoms of illness as well as female rats were excluded from this study.

After two weeks of acclimatisation, all 16 rats were randomly divided into two groups of eight rats each.

Group I: Control, received normal saline.

Group II: Arsenic (as sodium arsenite at the doses of 50 PPM once in a day for 90 days through oral gavage).

Study Procedure

The dose of arsenic was chosen according to a previously published study (24). Because of paucity in behavioural parameters and duration of exposure (90 days), the dose of present study was chosen on previously published report because the experimental animal model start appearing number of clinical and biochemical manifestation after 8-10 weeks of exposure.

Chemical: Sodium arsenite (NaAsO2), purchased from LOBA Chemicals, was used in this study. All other reagents used in this experiment were of analytical grade and obtained from commercial sources.

Experimental animals: Male Sprague-Dawley rats free of specific pathogens were purchased from Biogen Attibele, Bangalore, India and kept in the central animal house of Sri Devaraj Urs Medical College, Kolar Karnataka, India. Rats were 10 to 12 weeks of age with a body weight varying from 150 to 180 grams and procured in well-maintained temperature ranging from 20°C-26°C with a 12 hours dark/light cycles. They were housed in polypropylene home cages confirming two rats per cage. Throughout the experimentation, rats were given free access to rodent feed pellets and water Ad-libitum.

Neurobehavioural Studies

After 90 days of treatment, all rats underwent behavioural tests. The test was performed in a separate room maintaining 20°C-26°C with noise and dust free room. Rats were acclimatised to the test room for 30 minutes prior to the testing. The tests were performed between 09:00 AM to 03:00 PM.

1. Open Field Test (OFT): OFT was used to assess locomotion and exploratory behaviours (25).

Equipment: Briefly, OFT consisted of an open-top box (90×90 cm) with 45 cm high walls to prevent the rats from escaping. The floor of arena was painted black. White lines were drawn on arena floor and divided into 36 square blocks and each block was 15 cm2 long. The central part consisted of four squares in the center of the apparatus and marked with a red line. Video camera and a 60 W white light bulb were fixed above the arena.

Procedure: Each rat was gently placed in the center of arena and allowed freely to explore the arena for 300 seconds. At the end of exploration, rats were transferred to their respective home cages. After each trails, arena was cleaned with 70% ethanol.

The following parameters were assessed (26):

a. Number of square line crossed
b. Number of rearing
c. Number of wall-rearing
d. Time spent in center
e. Freezing time
f. Number of grooming

Note: Line crossed was only considered if the rats cross one of the lines with all four paws. Freezing time was only considered if the rats remain static for more than six seconds.

2. Beam Walking Test (BWT): Motor coordination and balance of rats were assessed by measuring the rat’s ability to traverse narrow beams to reach an enclosed safety platform (27).

Equipment: The Beam consisted of 150 cm long strips of wood, stretched between a blind and home end. It was 5 cm and 2.5 cm square thick, placed 50 cm above the floor. Just below the beam, a soft cloth was placed to prevent rats from injury in case of fall from the beam during experiment. An aversive stimulus (60 W light) was positioned near the blind end of the beam (27).

Procedure: Rats were individually kept at the blind end and allowed to walk on the beam. The animals’ performances were recorded with a video-camera. After each trails, test apparatus was cleaned with 70% alcohol to remove any smell and residue.

The following parameters were assessed:

a. Time taken to traverse the beam
b. Number of foot slips
c. Number of near fall
d. Actual fall

Note: All neurobehavioural tests were assessed three times and mean of these values was taken.

Sample collection: Following the behavioural tests, all rats were fasted over-night except water. Rats were deeply anaesthetised by injecting Ketamine (92 mg/kg Body-weight) and Xylocaine (10 mg/kg Body-weight) intraperitoneally and sacrificed by cervical decapitation. Blood was drawn from retro-orbital plexus and collected in heparinised vials, centrifuged to collect serum and stored in -80oC until use. Brain was removed, rinsed in cold saline and cerebellum was separated quickly. Half of the cerebellum was quickly frozen in liquid nitrogen and stored at -80°C and used for biochemical analysis. Remaining half was stored at -80°C and used for arsenic estimation.

Estimation of Arsenic Concentration in Serum and Cerebellum of Rat

Arsenic concentration in serum and cerebellum were assessed according to the procedure reported by Ballentine R et al., (28). A 200 mg of cerebellum was added in di-acid mixture which contains HNO3 and HClO4 in the ratio of 10:4, respectively. The sample was digested over a sand bath fume-hood chamber until a white gelatinous residue was formed. It was cooled and brought to 20 mL of volume by adding distilled water. Aliquots of digested and undigested samples such as cerebellum and serum were used to estimate arsenic using Atomic Absorption Spectrophotometer (Thermo-Scientific iCE 3000 series AA Spectrophotometer). The values are expressed in μg/g for tissue and ng/mL in serum.

Preparation of tissue homogenate for biochemical parameters: Each cerebellar tissue was homogenised individually in phosphate buffer Solution (pH 7.4) by using Glass-Teflon homogeniser. The cerebellar tissue segments were cut and fabricated in a 1:10 W/V (i.e., 1 gram of tissue in 10 mL of PBS) and homogenised for 10-15 minutes at 4°C. The homogenate was centrifuged at 15000 xg for 30 minutes at 4°C to remove debris. Clear supernatant was taken and stored at -80°C (29). The supernatant obtained from the tissues was used to estimate biochemical parameters such as Malondialdehye, Nitric Oxide and Glutathione Peroxidase spectrophotometrically (Table/Fig 1) (30),(31),(32).

Statistical Analysis

Statistical Package for the Social Sciences (SPSS) IBM, version 22.0 was used for analysis. Mean±SD was used for normally distributed data and the significance of difference was tested by independent t-test. Non normally distributed data were represented as Median (25th-75th Percentiles). Mann-Whitney U test were used for non parametric variables to check the difference in median between the groups. p-value <0.05 was considered statistically significant.

Results

General toxicity: There was alteration in eating, drinking and defecation. Following three to five days of treatment with arsenic, we noticed a reduction in food and water intake for 15-20 days, thereafter consumption of food and water was almost similar compared to controls. After 45 days till decapitation, they reduced food and water intake. A few changes were also noticed in arsenic treated rats like alteration in texture of stool, bloated abdomen, restlessness.

Body weight, brain weight and arsenic concentration: Arsenic treated rats showed significant reduction in the body weight gain after 90 days when compared to body-weight gain of control rats after 90 days. There was also significant decrease in brain weight in arsenic-exposed rats compared to controls (p-value <0.001). Group II also increased arsenic concentration in serum and cerebellum of rats (Table/Fig 2).

Oxidative Stress (OS) markers: To check the impact of NaAsO2 on OS- parameters, the activity of GPx and concentration of MDA and NO in serum and cerebellum of rats were assessed (Table/Fig 3).

The current results documented that rats treated with arsenic (Group II) significantly increases the concentration of MDA and NO and decreased the activity of GPx in serum and cerebellum of rats in comparison with control rats. The neurobehavioural activities were measured in OFT and BWT in experimental rats.

Effect of arsenic on OFT parameters: OFT was performed to assess the effect of oral administration of NaAsO2 on locomotion and exploratory behaviour. Group II rats significantly reduced the frequency of square line crossed, rearing, wall rearing and no of grooming in comparison with group I. On the other hand, freezing time increased significantly in group II, but there was no difference in grooming duration between the groups (Table/Fig 4).

Effect of arsenic on Beam Walking Test (BWT) parameters: BWT was done to assess the effect of oral administration of NaAsO2 on motor co-ordination and balance of rats (Table/Fig 5)a,b.

The current study indicated that group II took longer time to traverse the beam and also experienced more number of foot slips and near fall in 2.5 cm and 5 cm thick beam compared to group I. This may suggest that NaAsO2 alters the motor co-ordination and balance.

All rats (control and arsenic treated) took more time to traverse 2.5 cm square thick beam as compared to 5 cm thick beam. And also number of foot slips and near falls was more in 2.5 cm thick beam as compared to 5 cm thick beam.

Discussion

There is an increasing concentration of arsenic in the environment due to rapid industrialisation and urbanisation, which contaminate the water, air and soil. The uses of arsenic contaminants are toxic to humans and animals even at low (<10 μg/L) concentration (33). The current finding reported that the administration of NaAsO2 causes an impact on neurobehavioural and neurochemical alterations in rat’s cerebellum.

The present results revealed that arsenic treated rats cause a reduction in body weight gain which is similar to the study conducted by Sárközi K et al., and Rodríguez VM et al., (34),(35). The deficit in body weight gain could be due to reduced food consumption or damage to the gastric mucosa that causes gastrointestinal alterations and may alter the texture of stool (36). The current study also distinguished that there was a significant reduction in brain weight which may indicate arsenic has crossed the BBB and altered the brain weight and changes in brain weight could indicate oxidative damage.

It is documented in animal experiments that arsenic can cross BBB and invade the brain parenchyma (20). In present study, arsenic concentration in serum and cerebellum significantly increased in arsenic-treated rats which might be a contributing factor to cerebellar neurotoxicity. The present study is parallel with the results of Guan H et al., and Markowski VP et al., (37),(38). However, the change in serum and cerebellum arsenic concentration in the current and previous study could be due to different durations and routes of exposure and also to different groups of rodents.

OS is considered a deciding factor that contributes and triggers the induction of arsenic toxicity and has solid relationship with accumulation of arsenic in many organs such as heart, kidneys, lungs, liver, muscles, spleen and brain. Its accumulation has a persistent potential to damage organs and tissues (39). To study the toxic signs caused by arsenic and its effect on oxidative stress, we assessed MDA, NO and GPx in serum and cerebellum.

The results of the present study indicated that NaAsO2 causes significant neurotoxicity as confirmed by increase in OS markers such as MDA and NO as well as decrease in GPx activity in the serum and cerebellum of rats. In the current study, the concentration of MDA and NO level increased significantly in cerebellum which indirectly reflects the body cells by the severity of free radicals attacks. The present results also points out a positive relationship between MDA and NO levels and arsenic concentration in serum and cerebellum of rats. Simultaneously, the oxidative damage parameters i.e., GPx found to be significantly reduced due to antioxidant enzyme consumption and the antioxidant enzyme; GPx had the ability to scavenge oxygen free radicals. These results may show that arsenic can induce oxidative stress in rats’ cerebellum. These findings are consistent with the earlier reports indicating an increase in MDA and NO level and decrease in GPx activity in the cerebellum (10),(40),(41).

Ingested arsenic accumulates in different targeted tissues and can interfere with many biochemical reactions and alter numerous physiological activities in various organs of body (42). To substantiate the motor coordination, balance, locomotion and exploratory behaviour in arsenic induced rats, we performed BWT and OFT.

Rats treated with NaAsO2 showed marked impairment in locomotor activities as evidenced by the decreased frequency of square line crossed and increased freezing time as addressed in OFT which was similar to the study conducted by Kaushal P et al., and Saritha S et al., (43),(44). The reduction in locomotion might be due to a cerebellar disorder because the cerebellum is especially linked to voluntary nervous system and can result improper coordination between the nervous system and the muscular junction and might also be due to the oxidative stress detected in the of arsenic treated cerebellum which decreased motility in open field (20),(34),(45). However, the present study data is contradictory to the study done by Bikashvili T et al., and stated that there was no difference in locomotion between the arsenic exposed and control rodents (46).

In addition, we also evaluated the exploratory activities and found that there was significant reduction in exploratory activities such as rearing and wall-rearing in arsenic-treated rats, which is parallel to the study conducted by Saritha S et al., (44). The reduction in rearing in arsenic treated rats may indicate weakening of the limb muscles and leads to motor impairment and balance of the body.

Damage to the cerebellar structures promotes impairment in motor tasks such as coordination of movements, balance, and timing to traverse the beam. The present study showed that NaAsO2 treated rats showed a significant impairment in fine motor co-ordination and balance by increasing time to cross the beam, number of foot-slips and near fall which may indicate arsenic induced cerebellar neurotoxicity. This is in consonance with the study done by Kim H et al., and stated that arsenic treated mice reduced the motor coordination compared to control (47). This impairment in motor dysfunction may be linked with cerebellar markers of lipid and DNA oxidation (45).

Limitation(s)

Less number of animals was used in each group, microscopic study and other biochemical parameters has not been evaluated, rats were given sodium Arsenite orally through oral gavage, hence further research can be carried out in different experimental animals with other routes of administration.

Conclusion

The oral administration of sodium arsenite (NaAsO2) at the doses of 50 PPM once in a day for 90 days showed reduction in body weight gain. The arsenic treated rats had high concentration of arsenic in serum and cerebellar tissue which indicates accumulation of arsenic in cerebellum of rats and may reduce the brain weight and probably could have increased the production of free radicals and might cause oxidative stress. The present study also confirms that reduction in locomotion and exploratory behaviours and impairment in motor coordination and balance which indicate arsenic induced cerebellar neurotoxicity. Further, molecular studies may confirm the cerebellar neurotoxicity which may strengthen the present study.

Acknowledgement

Authors would like to thank Dr. Sarala N Department of Pharmacology and Incharge animal house facility, Sri Devaraj Urs Medical College, Kolar for providing animal house to carry out the study. Also would like to thank Mr. Ravi Shankar, Statistician, Department of Community medicine, for his timely help.

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DOI and Others

DOI: 10.7860/JCDR/2022/53048.16157

Date of Submission: Oct 28, 2021
Date of Peer Review: Nov 30, 2021
Date of Acceptance: Jan 25, 2022
Date of Publishing: Mar 01, 2022

AUTHOR DECLARATATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? NA
• For any images presented appropriate consent has been obtained from the subjects. NA

PLAGIARISM CHECKING METHODS:
• Plagiarism X-checker: Oct 31, 2021
• Manual Googling: Jan 19, 2022
• iThenticate Software: Feb 18, 2022 (7%)

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