Institutional Animal Ethics Committee Research Articles

Ameliorative effect of Curcuma longa against arsenic induced reproductive toxicity in Charles Foster rats

The present study aims to develop the antidote against arsenic induced male reproductive toxicity in animal models. The study was carried out on Charles Foster Rats after the approval from Institutional Animal Ethics Committee. A total of n=18, rats (12 weeks old) of an average weight of 160 ± 20 g were used for the study. The study group included n=6 control and n= 12 treated with sodium arsenite orally at the dose of 8mg/Kg b.w daily for 40 days. The n= 6 animals were dissected and rest n=6 was administered orally with Curcuma longa rhizome ethanolic extract at the dose of 600mg/Kg b.w per day for 40 days. At the end of the entire experiment, all the animals were dissected out and their reproductive organs were taken out, especially epididymis for sperm counts, sperm motility, sperm mortality, sperm morphology. The blood samples were collected for the hormonal assay (testosterone and luteinizing hormone), as well as for hematological and biochemical analysis. The study showed, high magnitude of degeneration in the reproductive organs of the rats in the arsenic treated group. There were degenerative fluctuations in the sperm counts, sperm motility, sperm mortality, sperm morphology and in the hormonal parameters, as well as in the hematological and biochemical parameters in the arsenic treated rats. But, after the administration of Curcuma longa, there was significant amelioration in all these parameters. Therefore, the present study shows that Curcuma longa plays vital role to combat arsenic induced male reproductive toxicity.

Mutagenicity and safety evaluation of Ashwagandha (Withania somnifera) root aqueous extract in different models

Withania somnifera (Ashwagandha) also called as Indian ginseng, a revered herb from Indian traditional system of medicine is a rejuvenator and tonic (Rasayana) used for its varied benefits. The roots of ashwagandha exhibit properties like anti-inflammatory, aphrodisiac, anthelmintic, astringent, diuretic, stimulant and thermogenic. However, data of ashwagandha on its mutagenic effects are lacking. In the present study, in-vitro genotoxicity tests were used to evaluate the mutagenic potential of Ashwagandha Root Extract (ARE). Concentrations of 0.156 to 5.00 mg/plate ARE were used for conducting Bacterial reverse mutation test (BRMT). For chromosome aberration (CA) test ARE was used in concentrations of 0.25 to 2.00 mg/ml, and for micronucleus (MN) tests ARE concentrations of 500/1000/2000 mg/kg were used. Acute oral toxicity was conducted in Wistar rats (n = 25) as per the OECD guideline (#423) with doses of 500/1000/2000 mg/kg body weight in male Swiss albino mice for morbidity and mortality for 3 days. The BRMT and CA tests were conducted with and without metabolic activation (S9). The study was approved by the institutional ethics committee (IEC) and institutional animal ethics committee (IAEC). ARE failed to show any mutagenic effects up to a dose of 5 mg/plate in BRMT. Also, ARE did not show any clastogenic activity in doses up to 2 mg/ml in CA test and in micronucleus test up to 2000 mg/kg body weight. These results were observed with and without metabolic activation (S9) under the stated experimental conditions. No mortality, morbidity, or any clinical signs were observed up to 3 days following ARE administration. Ashwagandha root extract failed to show any mortality in doses up to 2000 mg/kg oral dosage and did not show any mutagenic (genotoxic) effects in high concentrations.

Evaluation of renal ischemia–reperfusion injury using CEUS in mice

BackgroundRenal ischemia–reperfusion injury (IRI) frequently occurs clinically. We investigated the value of contrast-enhanced ultrasonography (CEUS) in the evaluation of renal IRI levels in mice.MethodsThirty-six healthy adult male C57BL/6 mice (20–22 g) were randomly divided into the sham, 10 min, 20 min, 30 min, 40 min, and 50 min groups based on the time of renal warm ischemia by blocking the left renal pedicle, approved by the Institutional Animal Ethics Committee. Time-intensity curve (TIC)-derived parameters such as peak enhancement (PE) and wash-in perfusion index (WiPI) were produced using CEUS at 1 h and 24 h after IRI. The severity of kidney injury was detected by the renal tubular necrosis rate which was analyzed by hematoxylin and eosin staining at 24 h after IRI. The Spearman correlation coefficient was used to describe the correlations between PE and WiPI values and the renal tubular necrosis rate.ResultsThe PE and WiPI values decreased after IRI in the groups with a warm ischemia time ≥ 20 min. The renal tubular necrosis rate was significantly correlated with the PE value at 1 h (ρ = -0.802) and 24 h (ρ = -0.861) after IRI and the WiPI value at 1 h (ρ = -0.814) and 24 h (ρ = -0.853) after IRI (all p < 0.001).ConclusionTIC-derived parameters, including PE and WiPI values, can be used to evaluate the severity of renal IRI in mice. CEUS is a safe and effective technology for the detection of renal IRI.Relevance statementCEUS can evaluate the severity of renal ischemia–reperfusion injury by peak enhancement and wash-in perfusion index values selected from various time-intensity curve-derived parameters.Key points• Contrast-enhanced ultrasonography can evaluate the level of renal ischemia–reperfusion injury.• Peak enhancement and wash-in perfusion index are correlated with the renal tubular necrosis rate.• CEUS can detect changes in unilateral renal function without radiation.Graphical

ANTI-INFLAMMATORY ACTIVITY OF SIDDHA POLYHERBAL FORMULATION SEVVIYADHI CHOORANAM ON CARRAGEENAN INDUCED PAW EDEMA IN WISTAR ALBINO RATS

Objective: The aim of the study was to explore the anti-inflammatory activity of Siddha polyherbal formulation Sevviyadhi chooranam in Carrageenan induced paw edema in wistar albino rats, and compared with the standard drug Indomethacin. Methods: The Siddha polyherbal formulation Sevviyadhi chooranam indicated for sinusitis was prepared based on GMP (Good Clinical Practice) guidelines. Study procedure was approved by Institutional Animal Ethics Committee (IAEC). The experimental animals were measured for paw edema volume at 1, 2, 3, 4, 5 h using Plethysmometer (Model 7150 UGO Basile, Italy). Edema was expressed as mean increase in paw volume relative to control animals. And then, findings were compared with Indomethacin (Standard drug). Results: The findings revealed that test drug Sevviyadhi chooranam at higher dosage 200 mg/kg (Group V) had equal effect on anti-inflammatory activity with percentage protection of 93.2% when compared with the standard drug Indomethacin at about 40 mg/kg (Group III) with percentage protection 93.2%. However, the test drug Sevviyadhi chooranam at a higher dosage 200 mg/kg (Group V) with a percentage protection 93.2% was highly effective when compared with lower dosage about 100 mg/kg (Group IV) with a percentage protection 27.12%. Hence, the study resulted that the Siddha polyherbal formulation Sevviyadhi chooranam has an optimistic anti-inflammatory activity with more therapeutic value. Conclusion: The study concluded that the Siddha polyherbal formulation Sevviyadhi chooranam has a promising anti-inflammatory activity, probably due to the presence of biologically active phytocompounds. However, it is important to admit that there are some scientific evidences of the potential actions of these phytocompounds in anti-inflammatory activity.

Protective effect of Cordia myxa leaves extract in FCA-induced arthritis in rats

Background: Arthritis, an age-related chronic inflammatory autoimmune disease primarily characterized by joint pain, swelling, stiffness, cartilage destruction, is increasing tremendously at an alarming rate and also becoming a major risk factor for other health complications. Cordia myxa, family Boraginaceae is an herbal plant used since ancient times for its various significant pharmacological activities, including anti-inflammatory effects. Scientific studies reporting anti-arthritic potential of leaves extract of this plant are still lacking. Therefore, the present work has been designed to investigate the protective effect of ethanolic extract of Cordia myxa leaves in Freund’s complete adjuvant (FCA)-induced arthritis in rats. Methods: Ethanolic extract (100mg/Kg, 200mg/Kg and 400mg/Kg) was investigated for their efficacy in the management of FCA induced arthritis in rats. Various parameters like paw volume (using plethysmograph), paw thickness (using Vernier calliper), body weight, hematological and histopathological profile were observed. The protocol was approved by Institutional Animal Ethical Committee (MMCP/IAEC/94) and was according to CPCSEA guidelines. Results: The phytochemical screening reveals the presence of flavonoids, glycosides, alkaloids, saponins and carbohydrates in the ethanolic extract of Cordia myxa leaves. Promising results with significant improvement were discovered as evident from all determined parameters when compared with standard drug methotrexate, which might be due to the presence of flavonoids, glycosides, alkaloids in the ethanolic extract of Cordia myxa leaves. Conclusion: The present study supports the traditional use of Cordia myxa leaves as potential anti-arthritic agent which can be effective in Freund’s complete adjuvant (FCA)-induced arthritis in rats, however its mode of action is still need to be studied.

CONSUMPTION OF HYPERLIPIDEMIC DIET AND WATER INTAKE IN ALBINO RATS TREATED WITH ECLIPTA ALBA – ORIGINAL RESEARCH PAPER

Objective: A high-fat (HF) diet described as higher calories generating from fat instead of carbohydrate or protein. HF usually diets a typical low-carbohydrate diet. Methods: Thirty adult Albino rats of Wistar strain accurately weighing between 165 and 215 g utilized for the present the study. As per CPCSEA guidelines, the protocol was approved by the institutional animal ethical committee no. BMCH/IAEC/01 Anat/2015 dated June 4, 2015. Animals were maintained and fed with standard rat chow, HF diet, and clean drinking water ad libitum, commercially available and manufactured by the National Institute of Nutrition, Hyderabad. Results: Rate of food consumption increased in rats fed with the HF diet for 8 weeks and decreased significantly in rats treated with Eclipta alba for 1, 2, and 3 weeks progressively compared to control group. Water intake increased in the rats fed with an HF diet (43±2.1) compared to rats fed with a normal diet, decreased significantly in the rats treated with E. alba (31±1.4) compared to the HF diet-fed animals. Conclusion: The rate of food consumption increased in rats fed with the HF diet for 8 weeks and decreased significantly in rats treated with E. alba for 1, 2, and 3 weeks progressively compared to control group. Water intake increased in the rats fed with an HF diet compared to rats fed with a normal diet, decreased significantly in the rats treated with E. alba compared to the HF diet-fed animals

Severity Classification of Laboratory Animal Procedures in Two Belgian Academic Institutions.

According to the EU Directive 2010/63, all animal procedures must be classified as non-recovery, mild, moderate or severe. Several examples are included in the Directive to help in severity classification. Since the implementation of the Directive, different publications and guidelines have been disseminated on the topic. However, due to the large variety of disease models and animal procedures carried out in many different animal species, guidance on the severity classification of specific procedures or models is often lacking or not specific enough. The latter is especially the case in disease models where the level of pain, suffering, distress and lasting harm depends on the duration of the study (for progressive disease models) or the dosage given (for infectious or chemically induced disease models). This, in turn, may lead to inconsistencies in severity classification between countries, within countries and even within institutions. To overcome this, two Belgian academic institutions with a focus on biomedical research collaborated to develop a severity classification for all the procedures performed. This work started with listing all in-house procedures and assigning them to 16 (sub)categories. First, we determined which parameters, such as clinical signs, dosage or duration, were crucial for severity classification within a specific (sub)category. Next, a severity classification was assigned to the different procedures, which was based on professional judgment by the designated veterinarians, members of the animal welfare body (AWB) and institutional animal ethics committee (AEC), integrating the available literature and guidelines. During the classification process, the use of vague terminology, such as 'minor impact', was avoided as much as possible. Instead, well-defined cut-offs between severity levels were used. Furthermore, we sought to define common denominators to group procedures and to be able to classify new procedures more easily. Although the primary aim is to address prospective severity, this can also be used to assess actual severity. In summary, we developed a severity classification for all procedures performed in two academic, biomedical institutions. These include many procedures and disease models in a variety of animal species for which a severity classification was not reported so far, or the terms that assign them to a different severity were too vague.

Pharmacological Evaluation of Ovulation Inducing Potential of Siddha Herbomineral Formulation Arputha Mathirai on Estradiol Valerate Induced Poly Cystic Ovarian Syndrome Wistar Albino Rat Model

Aim: The aim of this study is to examine the potential of Arputha Mathirai, a tablet-based Siddha herbomineral formulation, in promoting ovulation in Wistar Albino rats with polycystic ovary syndrome (PCOS) induced by Estradiol Valerate (EV).&#x0D; Place of Study: The study took place at C.L. Baid Metha College of Pharmacy located in Thorapakkam, Chennai - 600 092, Tamil Nadu.&#x0D; Methodology: Arputha Mathirai, the Siddha herbomineral formulation, was prepared in accordance with Good Manufacturing Practices (GMP) guidelines. Prior to conducting the study, approval was obtained from the Institutional Animal Ethics Committee (IAEC). The research adhered to ethical principles and guidelines established by the committee responsible for overseeing and regulating animal experimentation, ensuring proper control and supervision.&#x0D; Female Wistar Albino rats were selected as the preferred rodent species for the study. Polycystic ovary syndrome (PCOS) was induced in the animals by administering subcutaneous injection of 100μg Estardiol valerate (EV). The reproductive cycles of the rats were synchronized to ensure consistency.&#x0D; The rats were divided into four groups, each containing six rats, as follows:&#x0D; &#x0D; Group I: Normal Control animals received 1 ml/kg of Sodium Carboxymethyl Cellulose (CMC) solution.&#x0D; Group II: Rats were orally administered Arputha Mathirai at a dosage of 100 mg/kg for 10 days.&#x0D; Group III: Rats were orally administered Arputha Mathirai at a dosage of 200 mg/kg for 10 days.&#x0D; Group IV: Received Clomiphene at a dosage of 10 mg/kg and served as the standard group.&#x0D; &#x0D; At the end of the study, blood samples were collected from the rats through retro-orbital and cardiac puncture. The levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol, and progesterone were estimated using ELISA method. Ovaries from the experimental rats were dissected out for histopathological studies.&#x0D; Results: The results consistently showed that the higher dosage of the drug (group III) had a more pronounced effect in normalizing hormone levels compared to the lower dosage group. However, it is important to note that the lower dosage group also exhibited effectiveness. While the results in both groups did not closely match those of the standard group (group IV), the drug Arputha Mathirai demonstrated significant potential in the experimental groups, supporting its efficacy in inducing ovulation.&#x0D; Furthermore, the histopathological analysis confirmed the drug's potential in inducing ovulation, particularly in the higher dosage group (group III) compared to the lower dosage group (group II).&#x0D; Overall, the findings suggest that Arputha Mathirai has promising ovulation-inducing activity, with the higher dosage showing more pronounced effects.&#x0D; Conclusion: It is hypothesized that the bioactive phytocompounds present in Arputha Mathirai may exert their effects at various stages to restore hormone levels and reverse the pathological condition associated with PCOS. These natural compounds have the potential to regulate hormone levels and promote ovulation. However, it is important to acknowledge that there is a scarcity of published evidence regarding the specific ability of these phytochemicals to address the underlying causes of the condition. Further research is needed to explore the mechanisms by which these compounds act and their potential in addressing the root causes of PCOS.

Catastrophic consequences of the enormous use of hydroxychloroquine during COVID era on liver and kidney of male albino rats: an in-vivo study

Background: Hydroxychloroquine (HCQ) is mainly used for the treatment of malaria but during COVID trial, it was used against coronavirus though no history of the drug is known against SARS COV 2 or any other respiratory ailment. Many case studies showed the adverse effects on liver and kidney in many patients after the exposure of HCQ. The main aim of this study is to know the effect of HCQ drug on the liver and kidney of male albino rat at a range of human equivalent dose that was given during COVID period. Methods: After institutional animal ethics committee (IAEC) approval, ten male albino rats were obtained and divided into two groups-control and treated. Treated groups receives HCQ through oral gavage for six days and then serum, tissue enzymes and total serum bilirubin were measured. Histopathological study was done from liver and kidney tissue. After that statistical analysis was done. Results: We found significant increase in enzymes glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and alkaline phosphatase (ALP) in the HCQ-treated rats than in control and this signifies that there might be damages that occurred in liver and kidney. Increased level of bilirubin in HCQ-treated rats indicate hyperbilirubinemia and may be a sign of jaundice or any other hepatic disorder. From histopathological identification we also found liver and kidney tissues got damaged due to exposure of HCQ. Conclusions: From this study, we can conclude that the exposure of this drug might have led to the impaired function of organs that could have potentiated their ill fate.

Ameliorative effect of aqueous extract of Carica papaya Linn. leaves on Acetic acid induced Ulcerative Colitis in Male Albino Wistar rats

Ulcerative colitis (UC) manifests as chronic inflammation of the colon. The bowel inflammation is due to alteration in the immune response to gut micro flora, oxidative stress and hereditary genetic factors. Leaf extract of papaya Carica papaya Linn. contain vitamin A, C, alkaloids, saponins, glycosides, tannins and flavonoids which have antispasmodic, analgesic, antibacterial and antioxidant properties. The main aim of the study was to detect the efficacy of Carica papaya leaf extract (PLE) on ulcerative colitis in Wistar Albino rats with objectives to explore the possible mechanism involved in its efficacy and to look for prebiotic activity. The project was initiated after the approval of Institutional Animal Ethics Committee. Thirty adult Wistar rats were randomly assigned into seven groups of six rats each. Group I-control, Group II-UC disease control, Group III-Disease with standard Sulfasalazine 100mg/kg, Group IV and Group V, UC disease model receiving PLE of 250mg/kg and 500 mg/kg respectively. Ulcerative colitis was not induced in Group VI and Group VII which received PLE of 500mg/kg and 700 mg/kg respectively. All the drugs were administered orally for 15 days. UC was induced by acetic acid on the 8th day. Blood sample was collected for biochemical analysis for antioxidant levels. Histopathological analysis of distal colon was performed to look for inflammatory features. Microbial analysis was done for group VI and group VII. There was significant reduction in body weight in UC induced groups when compared to control. The weight loss in Group IV and V (PE1 and PE2) was not as extreme as seen in the UC alone group. The treatment groups IV and V showed significant increase in protein levels, increased catalase and decreased malondialdehyde levels when compared to Group I, II and III. On histopathological analysis, the severity and extent of inflammation was less in 500mg/kg dose of leaf extract (Group V) than 250mg/kg of leaf extract (Group IV) and 100mg/kg of sulfasalazine (Group III). Tissue sections of Group V showed only few inflammatory infiltrates with absence of oedema and crypt abscess in the submucosal area. On examining the pathogenic versus beneficial bacterial load in the control and Group VI (PE 3) and Group VII (PE 4), papaya leaf extracts depicted antibacterial property and probiotic activity. In the present study, 500mg/kg of papaya leaf extract was efficacious in relieving oxidative stress and reducing inflammation in acetic acid induced ulcerative colitis in rats. Carica papaya leaf extracts have a potential role to modify gut microbe with respect to its prebiotic activity.

The Use of Retraction Sutures in End-To-Side Microvascular Anastomosis—A Novel Technical Innovation in Experimental Rat Femoral Model with Successful Use in STA-MCA Bypass in Moyamoya Disease

A narrow working space, small diameters, and the tendency to collapse with clamps make cerebral microvascular anastomosis challenging. A retraction suture (RS) is a novel technique to keep the recipient vessel lumen open during the bypass. To provide a step-by-step overview of RS for end-to-side (ES) microvascular anastomosis on rat femoral vessels and successful use for STA-MCA bypass in Moya Moya disease (MMD) patients. A prospective experimental study with approval from the Institutional Animal Ethics Committee. Femoral vessels ES anastomoses were performed on Sprague Daley rats. The rat model used three types of RS (Adventitial, Luminal, and Flap-retraction sutures). An ES-interrupted anastomosis was done. The rats were observed for an average period of 16.18±5.65 days; the patency was assessed by re-exploration. The immediate patency on the STA-MCA bypasses was confirmed with intra-operative indocyanine green (ICG) angiography and micro-doppler; delayed patency with MRI and DSA after 3-6 months. In the rat model, 45 anastomoses were performed, 15 each using the three subtypes. The immediate patency was 100%. Delayed patency was 42/43 (97.67%), and two rats died during observation. In the clinical series, 59 STA-MCA bypasses were done in 44 patients (average age -18.14±11.09 years) using RS. The follow-up imaging was available for 41/59 patients. Both immediate and delayed patency (41/41 at six months) was 100%. The RS allows continuous visualization of the vessel lumen, reduces the handling of intimal edges, and avoids incorporating the back wall in sutures; thus improving anastomosis patency.

Assessment of Nephroprotective Activity of PippalyadiAgad on Diclofenac Sodium Induced Nephrotoxicity in Albino Mice: An Experimental Study

Background: Nephrotoxicity due to Diclofenac Sodium can be correlated with this concept of Dushi Visha and nephroprotective drugs mentioned in Ayurveda texts may play an important role in this area. Pippalyadi Agad is the herbo-mineral medication stated by Yogaratnakara for treatment of Dushi Visha. Some of the drugs in this formulation have shown efficacy in correcting nephrotoxicity. Objective: To study the nephroprotective activity of Pippalyadi Agad on diclofenac sodium induced nephrotoxicity in albino mice. Methods: The experimental study was carried out in albino mice as per CPCSEA guidelines after getting approval from the Institutional Animal Ethics Committee (IAEC). 30 albino mice, each weighing about 20-30 gm, were procured from Bharat serum and vaccines. The mice were housed in the institutional animal house in standard conditions. 30 albino mice were equally divided into five groups having six animals in each group as Trial Drug (low dose), Trial Drug (medium dose), Standard Control, Disease Control and Vehicle Control. For assessment of nephrotoxicity, the animals were monitored on Day 0, Day 15 and Day 45 by carrying out blood investigations such as BUN, Serum Creatinine, Body weight and CBC. The Data were subjected to appropriate statistical tests to derive results. Results: Pippalyadi Agad treatment at low and medium doses significantly lowered BUN and Serum Creatinine in comparison with Disease and Standard Control at Day 45. Pippalyadi Agad treatment at low and medium doses significantly (p &lt; 0.001 and p &lt; 0.001) increased body weights of the animals in comparison with Disease and Standard Control at Day 45. No significant difference in CBC parameters was observed in any of the Groups except the Disease Control Group on Day 15 and Day 45. Conclusion: Pippalyadi Agad has shown nephroprotective activity based on biochemical changes observed in Serum Creatinine, BUN and CBC.

Protective Effect of Lycopene Rich Tomato Puree on the Development of Atherosclerosis/Heart Diseases in Animal Models

Background: Lycopene as an aliphatic carotenoid promulgated from tomato has gained the attention of new generation food and pharma scientists on the ground of its nutraceutical efficacy. It is a preeminent antioxidant known for its singlet oxygen quenching ability that reduces oxidative stress responsible for non-communicable diseases. In present investigation the health claim exploration ability of lycopene rich tomato puree as a dietary supplement was assessed by animal studies to project its hypolipidemic efficacy effective in reducing risk of heart diseases.&#x0D; Aim: The present investigation aims to determine efficacy of tomato puree lycopene in monitoring atherogenesis mechanism through synergic mode of action of notified specialty features (total cholesterol, serum triglycerides, LDL cholesterol and HDL cholesterol) in Wistar rats.&#x0D; Study Design: Wistar rats (Positive control, high fat diet fed, HFD-Stain fed and HFD-tomato puree fed) were examined for general examinations (behavioural changes and body weight), Lipid profile (total cholesterol, triglyceride, LDL and HDL cholesterol) on 0, 14 and 28th day of experiment to assess effect of tomato puree administration on wellbeing of rats against healthy, hyperlipidemic and standard drug status.&#x0D; Place and Duration of Study: MIT School of Food Technology in collaboration with Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Parbhani (MAFSU, Nagpur), Maharashtra. The overall study duration was for 3 months.&#x0D; Methodology: The experimental rats (Wistar) employed in present study were acquired from authorized breeder after the approval of the experimental protocol by Institutional Animal Ethical Committee (IAEC approval resolution no.: IAEC 58/19, Date 20/08/2019). Acclimatization of the animals was undertaken for 20 days during which they were provided with natural diet, pure and safe drinking water. CPCSEA (Committee for the purpose of control and supervision of experiments on animals) guidelines were followed while handling the experimental animals throughout the complete study duration. The experimental animals (32 Wistar rats) were segregated into four different groups (Positive control, high fat diet fed, HFD-Stain fed and HFD-tomato puree fed), containing 8 rats (4 male and 4 female). Animals from four distinguished groups were monitored regularly for general observations (body weights, behavioural changes and mortality changes) throughout the experimental period. The lipid profile (total cholesterol, triglyceride, HDL, LDL) of blood serum samples was recorded using auto analyser on 0, 14th and 28th day of experiment.&#x0D; Results: The data analysis, interpretation and reviewed literature infusion collectively underlined the efficacy of lycopene (tomato puree) in monitoring atherogenesis mechanism through synergic mode of action of notified specialty features (total cholesterol, serum triglycerides, LDL cholesterol and HDL cholesterol) and projected an effective gateway to support disease infused health claim. The cumulative decrease in total cholesterol (159.99 to 126.40mg/dl), serum triglycerides (114.81 to 107.34mg/dl), LDL cholesterol (40.19 to 32.90mg/dl) and increase in HDL cholesterol (19.99 to 23.87mg/dl) of treated rats represents hypolipidemic efficacy of tomato puree lycopene.&#x0D; Conclusion: The tomato puree mass being a viable lycopene source with innate antiatherogenic efficacy has rationally updated the review base critical appraisal leading to development of prototype of CVD to launch health claim. The human clinical study as a confirmation tool, may be the best option for next generation research.

Effect of Bezafibrate, a PPARα Activator, on Acute and Subacute Inflammation in Male Wistar Rats

Aim To investigate the anti-inflammatory effect of bezafibrate on acute and subacute inflammation in adult male Wistar rats. Methods The study was carried out in adult male Wistar rats and they were allocated into three groups, that is, control, aspirin and bezafibrate, after obtaining clearance from Institutional Animal Ethics Committee. Acute inflammation was studied using carrageenan-induced rat paw oedema, and the volume displacement due to paw oedema using the plethysmograph. Subacute inflammation was studied using foreign body induced granuloma models. Analysis was performed using one-way ANOVA followed by post hoc tests of Dunnett’s. The value p &lt; 0.05 was considered statistically significant. Results Bezafibrate showed a significant anti-inflammatory effect in acute as well as subacute models of inflammation when compared to control in the present study. Conclusions In patients receiving bezafibrate for hyperlipidemia, its anti-inflammatory potential may have an additional benefit in preventing complication of atherosclerosis.

1146 SLEEP DEPRIVATION INDUCES AGEING-LIKE CHANGES IN ANTIGRAVITY MUSCLES OF YOUNG ADULT MALE WISTAR RATS

Abstract Introduction Poor muscle health is associated with a series of chronic and metabolic conditions that are prevalent in individuals who chronically experience poor-quality sleep. But there is no study deciphering the role of sleep deprivation on muscle ageing. Therefore, in the present study we have measured the ultrastructure, histopathology, and oxidative stressors in soleus muscle of wistar rat after sleep deprivation and recovery sleep. Material and Methods The experiments were conducted in 18 rats of three groups. Group I rats had normal sleep wake cycle, Group II rats were subjected to 24 h sleep deprivation (SD) by gentle handling method1 and Group III rats had recovery sleep after 24 h SD. At the end of the sleep, sleep deprivation and recovery period, soleus muscle tissue was collected for ultrastructural, histological and oxidative stress markers. Oxidative damage was assessed by lipid peroxidation, catalase activity, reduced glutathione and nuclear labelling of 8-OHdG. The study was conducted as per the guidelines of the Institutional Animal Ethics Committee (960/IAEC/16). Results The data demonstrated that SD leads to ultrastructural changes in soleus muscle which includes sarcolemmal and mitochondrial alterations. In case of histopathological and histomorphological changes there was signs of tissue degeneration, inflammatory infiltrate in type I fibres and muscle atrophy was observed in soleus muscles. There was significant increase in level of 8-OHdG (p=0.02) and malondialdehyde in 24h SD (p=0.02) than control and recovery sleep groups. Moreover, the catalase activity and reduced glutathione level was significantly decreased in 24h SD group (p≤0.02) than control and recovery sleep. Conclusion 24hr sleep deprivation leads to an ageing like state in the skeletal muscle, which was recovered after sleep rebound.

Dehydroepiandrosterone and Acute Stress Attenuation: An Interventional Rodent Study

Introduction: Stress activates hypothalamo-pituitary-adrenal axis leading to the release of glucocorticoid that mediates the stress response. This adaptive response is self-limited but if persistent for prolonged periods can lead to disease states. Nature has endowed the body with efficient buffer systems to attenuate the stress effects and Dehydroepiandrosterone (DHEA), a steroid hormone with neuromodulatory functions is implicated as an efficient candidate to buffer stress. Aim: To assess the effect of prophylactic administration of DHEA in the attenuation of acute stress in male Wistar rats. Materials and Methods: This interventional study was carried out at centre for Toxicology and Developmental Research, Sri Ramachandra Institute of Higher Education and Research, Chennai, between June 2021 and August 2021, in compliance with the animal welfare guidelines of CPCSEA, and in accordance to the protocol approved by Institutional animal ethics committee. The 18 male Wistar rats approved for the study were segregated into 3 groups with 6 animals in control (no stress) group, 6 in stress group and 6 in intervention group that received DHEA prophylactically 30 min before stress procedure. Animals in stress and intervention groups were subjected to one hour immobilisation stress. Blood samples were collected from all animals after the stress period and serum corticosterone, the stress marker, was estimated. The data were expressed as mean±standard error of mean (mean±SEM) and Mann-Whitney U test was used to test the significant difference between the: (i) control &amp; stress groups; (ii) stress &amp; study groups; and (iii) control &amp; study groups. The p-value&lt;0.05 was considered significant. The analysis was done using SPSS version 23.0. Results: The values of corticosterone in control, stress and intervention groups were 26.6±4.4 ng/mL, 51.6±3.9 ng/mL and 23.4±3.6 ng/mL, respectively. Significant difference in the mean serum corticosterone levels with p-value 0.013 between control and stress groups and with p-value 0.008 between stress and DHEA groups were observed. Conclusion: It could be observed from the findings that prophylactic DHEA administration attenuated acute stress efficiently in male Wistar rats as reflected by the significant decrease in serum corticosterone levels in the group that received DHEA intervention, thus inferring the efficiency of DHEA in stress buffering.

Diagnostic Tests for Confirmation of Mating in Adult Female Wistar Rats: An Experimental Study

Introduction: The assessment of confirmation of mating in rats is a procedure which requires expertise with caution, and careful execution. Rats are spontaneous ovulators and are widely used to study the reproductive system. Aim: The aim of the present study study was to compare diagnostic tests for confirming mating in adult female wistar rats. Materials and Methods: The present experimental research was duly approved by the Institutional Animal Ethical Committee and conducted at the Centre for Toxicology and Developmental Research (CEFTE) at Sri Ramachandra Institute of Higher Education and Research (Deemed to be University). The study was conducted in December 2019. A total of 72 adult Wistar rats (36 males and 36 females) were used in the study. To identify the event of mating, the rats were subjected to the following confirmatory methods: naked eye examination, non invasive vaginal lavage method, and abdominal palpatory method. Data were subjected to statistical analysis using non parametric tests with SPSS software (version 25.0). The level of significance was set at p&lt;0.05. Comparison of all three methods was done using non parametric tests (post hoc with Wilcoxon signed-rank test). Results: The non invasive vaginal lavage method was found to be more accurate (72.22%) compared to the visual method (16.66%) and the abdominal palpatory method (11.11%). A significant difference was seen when comparing the three methods using post hoc with Wilcoxon signed-rank test (p≤0.05). Conclusion: Based on the findings, it can be concluded that the vaginal lavage method is more specific and easy to confirm the occurrence of mating in adult female Wistar rats. The non invasive vaginal lavage method is a simple, easy, and accurate method for estimating pregnancy in cohabitating/generation studies.

Therapeutic Effect of Tinospora cordifolia (Willd) Extracts on Letrozole-Induced Polycystic Ovarian Syndrome and its Complications in Murine Model.

Tinosopora cordifolia (Willd) (TC) is commonly used in Ayurvedic medicine since long time for number of ailments and its preparations are also considered by food safety and standards authority of India as nutritional supplement. However the scientific evidence on its possible safety and efficacy in polycystic ovarian syndrome and associated complications was not studied in detail. The purpose of this investigation is to examine whether or not TC can have therapeutic effects on letrozole induced PCOS and related complications such as body weight, dyslipidaemia, glucose tolerance, hormonal regulation, insulin resistance and sensitivity, severity of PCOS and histopathological changes in ovary using mice animal model. Present study is a preclinical study involving laboratory animals. After verifying the absence of PCOS, the animals began receiving Letrozole, which lasted for 21 days. Fasting blood glucose (FBG), the oral glucose tolerance test (OGTT), triglycerides, cholesterol, and weight were recorded. The levels of hormones like oestrogen, progesterone, insulin, testosterone, luteinising hormone (LH) and follicle stimulating hormone (FSH), histopathology was carried out. The Institutional Animal Ethics Committee at DITU gave its clearance to the animal experimentation on July 10, 2021 (DITU/IAEC/21-22/07-06). The majority of cornified epithelial cells were seen in groups treated with TC extract during the estrous phase of the cycle. Mice exposed to TC retained normal body weight. FBG, 1- and 2-hour OGTT, triglyceride and cholesterol levels were all significantly improved by extracts. Estradiol, progesterone, testosterone, insulin, LH and FSH concentration were all corrected in TC-treated animals. The HOMA-IR, HOMA-Beta and QUICKI values were also corrected with TC extracts. The morphological and microscopic features of the ovary were also greatly enhanced. Based on these findings, we conclude that treating PCOS mice with TC extracts significantly ameliorates the disease and severity down to nil-to-moderate levels by reducing hyperinsulinemia, hyperandrogenism, dyslipidaemia, enhancing insulin sensitivity, correcting oestrogen, progesterone, LH and FSH levels via enhanced ovarian function. Further molecular and cellular level of study is recommended for further elaboration of mechanism of action. • Tinospora cordifolia satva, oil and hydroalcoholic extract were studied in letrozole-induced PCOS in mice model• Anti PCOS efficacy of 3 preparations studied with respect to their mechanism of action in detail• For the first time proposing method of calculating severity of PCOS in animal model• Tinospora cordifolia oil preparation completely reversed PCOS effect of letrozole and made them normal• Histopathological and morphological studies support the biochemical claims.

Histological and immunohistochemical effects of chronic intraperitoneal monosodium glutamate at doses equivalent to human consumption in cerebellar cortex of the adult mice

Background: Monosodium glutamate (MSG), a flavor enhancer, is one of the most widely used commercial food additives. MSG produces oxidative stress in various tissues including cerebellar cortex. There is a paucity if data on the histological and immunohistochemical changes of glial fibrillary acidic protein (GFAP) on chronic administration of MSG at doses equivalent to human consumption (i.e., 40 mg/kg, 60 mg/kg, and 80 mg/kg daily for 3-month duration), hence this study. Aims and Objectives: The aim of the study was to evaluate the histological and immunohistochemical effects produced by MSG at doses equivalent to human consumption (40mg/kg/day, 60 mg/kg/day, and 80 mg/kg/day) in cerebellar cortex of adult mice. Materials and Methods: Study commenced after the Institutional Animal Ethics Committee approval. 40 Swiss-albino mice were randomly divided into four groups. Group 1 served as the negative control and received distilled water i.p. Groups 2, 3, and 4 received MSG at increasing doses. Animals were euthanized at 3 months, cerebellar samples were examined histopathologically and immunohistochemically. Results: Degeneration of the layers of cerebellar cortex in histopathology and reduction in the GFAP activity was observed in immunohistochemistry. Conclusion: MSG due to its wide consumption can cause degenerative changes in the cerebellum.

An in vivo and in silico evaluation of the hepatoprotective potential of Gynura procumbens: A promising agent for combating hepatotoxicity.

The liver, the most important metabolic organ of the body, performs a wide variety of vital functions. Hepatic cell injury occurs by the activation of reactive oxygen species (ROS) that are generated by carbon tetrachloride (CCl4), xenobiotics, and other toxic substances through cytochrome P450-dependent steps resulting from the covalent bond formation with lipoproteins and nucleic acids. Observing the urgent state of hepatotoxic patients worldwide, different medicinal plants and their properties can be explored to combat such free radical damage to the liver. In vivo and in silico studies were designed and conducted to evaluate the antioxidant and hepatoprotective properties of Gynura procumbens in rats. Gynura procumbens leaves were collected and extracted using 70% ethanol. The required chemicals CCl4, standard drug (silymarin), and blood serum analysis kits were stocked. The in vivo tests were performed in 140 healthy Wister albino rats of either sex under well-controlled parameters divided into 14 groups, strictly maintaining Institutional Animal Ethics Committee (IEAC) protocols. For the histopathology study, 10% buffered neutral formalin was used for organ preservation. Later the specimens were studied under a fluorescence microscope. In silico molecular docking and absorption, distribution, metabolism, excretion, and toxicity (ADMET) studies were performed, and the results were analyzed statistically. Gynura procumbens partially negate the deleterious effect of carbon tetrachloride on normal weight gain in rats. The elevated level of serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), creatinine, LDH, total cholesterol (TC), low-density lipoprotein (LDL), triglycerides (TG), malondialdehyde (MDA), deoxyribonucleic acid (DNA) fragmentation ranges, gamma-glutamyl transferase (γ-GT) in CCl4 treated groups were decreased by both standard drug silymarin and G. procumbens leaf extract. We have found significant & highly significant changes statistically for different doses, here p<0.05 & p<0.01, respectively. On the other hand, G. procumbens and silymarin displayed Statistically significant (p<0.05) and high significant(p<0.01) increased levels of HDL, CAT SOD (here p<0.05 & p<0.01 for different doses) when the treatment groups were compared with the disease control group. Because the therapeutic activity imparted by plants and drugs accelerates the movement of the disturbed pathophysiological state toward the healthy state. In the molecular docking analysis, G. procumbens phytoconstituents performed poorly against transforming growth factor-beta 1 (TGF-β1) compared to the control drug silymarin. In contrast, 26 phytoconstituents scored better than the control bezafibrate against peroxisome proliferator-activated receptor alpha (PPAR-α). The top scoring compounds for both macromolecules were observed to form stable complexes in the molecular dynamics simulations. Flavonoids and phenolic compounds performed better than other constituents in providing hepatoprotective activity. It can, thus, be inferred that the extract of G. procumbens showed good hepatoprotective properties in rats.