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ThesisItem Open Access BIOENHANCING EFFECT OF BERBERIS ARISTATA AND CURCUMA LONGA ON PHARMACOKINETICS OF AMOXICILLIN IN CALVES(CSKHPKV, Palampur, 2014-07) Tandon, Tamanna; Varshneya, C.The bioenhancing effects of Berberis aristata and Curcuma longa (30 mg.kg-1) were studied on pharmacokinetics of amoxicillin in calves. The studies were carried out on six apparently healthy male calves (2-3 months of age). The calves were divided into six groups and amoxicillin was administered at the dose rate of 10 mg.kg-1 by intravenous and oral route in groups I and II, respectively. The animals of group III and IV received extracts of Berberis aristata and Curcuma longa along with amoxicillin, respectively. The animals of group V and VI received only the extracts of Berberis aristata and Curcuma longa, respectively. The blood samples were collected at 2.5 min., 5 min., 10 min., 15 min., 30 min., 1h, 2 h, 4 h, 6 h, 8 h, 10 h and 12 h following the intravenous administration and samples were collected at 15 min., 30 min., 1h, 2 h, 3 h, 4 h, 5 h, 6 h, 8 h, 10 h and 12 h after oral administration. The samples were analysed for amoxicillin using agar plate diffusion method. The disposition kinetics of amoxicillin followed two compartment open model following intravenous and oral administration. Following a single intravenous injection of amoxicillin, the plasma level was 11.9 ± 0.21 μg.ml-1 at 2.5 min. and the drug was detected above MIC in plasma upto 6 h. The pharmacokinetic parameters of the distribution and elimination phase of amoxicillin revealed rapid distribution of the drug into body fluids and tissues with a short distribution half life of 0.13 ± 0.01 h. A fair distribution of drug was also reflected by the high values of K12/K21 ratio (1.81 ± 0.05) and AUC (16.13 ± 0.28 μg.ml-1.h). The values of t1/2β, Kel, ClB and MRT were: 2.50 ± 0.07 h, 0.88 ± 0.01h-1, 620.75 ± 10.49 ml.kg-1.h-1 and 3.18 ± 0.08 h, respectively. After oral administration the peak plasma level was achieved at 3 h and the values of AUC, t1/2β, ClB and MRT were: 9.25 ± 0.31 μg.ml-1.h, 1.66 ± 0.05 h, 617.03 ± 10.71 ml.kg-1.h-1 and 3.79 ± 0.06 h, respectively. The simultaneous administration of extracts of Berberis aristata and Curcuma longa altered both the absorption and elimination phases. The findings of the present study revealed that amoxicillin has a bioavailability of 57 percent. The simultaneous administration of extracts significantly (P<0.05) increased the oral bioavailability of amoxicillin.ThesisItem Open Access STUDIES ON AMELIORATIVE POTENTIAL OF SEABUCKTHORN AGAINST AFLATOXIN INDUCED HEPATOTOXICITY IN POULTRY(CSKHPKV, Palampur, 2014) Sharma, Navjot; Varshneya, C.The present investigation was undertaken to study the ameliorative potential of seabuckthorn (leaves and pulp) in aflatoxin induced hapatotoxicity in poultry. A total of 210, day old broiler chicks were used for studying the growth performance, biochemical parameter and antioxidant status for the period of 4 weeks. 70% methanolic extracts of seabuckthorn leaves and pulp were fed to broiler chicks @ 1000ppm in feed . Glucomannan was mixed in feed @ 1g/kg whereas, aflatoxin was added @ 400ppb. The aflatoxin treatment significantly (P<0.05) reduced weight gain and increased FCR. The seabuckthorn (leaves and pulp extract) alone significantly (P<0.05) improved the growth performance. AF treatment produced a significant (P<0.05) decrease in total serum proteins, albumin, globulin, cholesterol, triglycerides and HDL and significant (P<0.05) increase in AST, ALT, ALP, LDH and total serum billirubin. However, seabuckthorn (leaves and pulp extract) and glucomannan supplementation significantly (P<0.05) restored these biochemical parameters to control levels. In aflatoxin treated groups the malondialdehyde (MDA) level of liver, kidney and blood was significantly (P<0.05) increased whereas, GSH, catalase, SOD and GPx levels were significantly (P<0.05) decreased, while there was a significant (P<0.05) increase in levels of MDA, GSH, catalase, SOD and GPx in liver of chicks fed on seabuckthorn. The combined dietary supplementation of AF + SBT leaves , AF + SBT pulp, and AF +GM produced significant (P<0.05) decrease in lipid peroxidation and an increase GSH, Catalase, SOD and GPx level of liver, kidney and blood, respectively. Histopathological lesions in the liver were severe in aflatoxin group with marked congestion of hepatic sinusoids, focal haemorrhages, centro-lobular fatty cytoplasmic vacuolation and or necrosis, biliary hyperplasia, severe degeneration of hepatocytes, dilation of central veins, bile duct proliferation and least severe in group (AF+GM) followed by groups fed with SBT (leaves and pulp extract) and there were no lesions in chicks fed with SBT (leaves and pulp) alone. On the basis of growth performance, biochemical parameters, antioxidant activity and histopathology of liver tissues, it can be concluded that dietary supplementation of seabuckthorn is capable of ameliorating the aflatoxin induced hepatotoxicity in poultry.ThesisItem Open Access PHARMACOLOGICAL & TOXICOLOGICAL EFFECTS OF PACLITAXEL(CSKHPKV, Palampur, 2014) Bhat, Mohammad Aamir; Varshneya, C.The present investigation was conducted to study in vitro cytotoxicity, apoptosis and changes in cell cycle kinetics induced by paclitaxel on C6 (Rattus norvegicus glioma) and CHO-K1 (Chinese hamster ovarian carcinoma) cell lines. In this study, percentage cytotoxicity against cell lines was evaluated by Sulphorhodamine (SRB) assay. The percentage of surviving cells fell after 48 hours of treatment and IC50 values were observed between 0.5 to 0.75 and 0.25 to 0.75 μg/ml in C6 and CHO-K1 cells, respectively. Cells incubated in high concentrations of paclitaxel had increased survivability compared with cells treated with lower concentrations of the drug. No significant cytotoxicity was observed after 24 hours of treatment. Paclitaxel induced apoptosis by caspase 3/7 activation and caused accumulation of cells in G2/M phase of cell cycle in both the cell lines. Upon fluorescent microscopy, both the cell lines lost their morphology, confluency and adherence after 24 hours but the effects were much more pronounced after 48 hours of treatment. In vivo toxicological studies were also conducted in adult wistar rats to evaluate the highest non toxic dose level of intra peritoneal administration of Paclitaxel-Cremophor EL formulation at 0.30 (G1), 1.20 (G2), 2.32 (G3) and 3.20 (G4) mg/kg at 7 day interval over a period of 21 days (3 times in total). In acute toxicity studies alopecia and tachypnea was observed in G4. No other signs of acute toxicity and no mortality was observed in any of the dose groups. After repeated dosing, at 21st day, serum aspartate transaminase, alanine transaminase, lactate dehydrogenase, creatinine phosphokinase levels were significantly increased in G3 and G4 and total serum protein, albumin and alkaline phosphatase levels were also increased in G4. No significant changes were observed in serum globulin levels in any of the dose groups. Red Blood cell and lymphocytic count was significantly decreased in groups G3 and G4 and white blood cell, neutrophil count, hematocrit percentage and haemoglobin concentration was also decreased in G4. Histopathological changes were observed mainly as nuclear fragmentation, apoptosis and cellular damage in liver, heart, spleen, kidney, mesenteric lymph nodes, lungs and testes in G3 and G4 but the severity of lesions was much higher in G4. The in vitro data suggested that paclitaxel will be most effective clinically when there is prolonged exposure of tumour to the drug rather than increasing concentration of drug in the biophase. Based on in vivo results, the highest non-toxic dose of formulation was estimated to be 1.20 mg/kg in rats, under this study condition and equivalent human dose was estimated to be 0.1332 mg/kg.ThesisItem Open Access ANTIOXIDANT AND IMMUNOMODULATORY ACTIVITY OF GLYCYRRHIZA GLABRA AND BAUHINIA VARIEGATA IN RATS(CSKHPKV, Palampur, 2014-07-18) Kanika; Varshneya, C.The present investigation was undertaken to study the antioxidant and immunomodulatory action of Glycyrrhiza glabra root and Bauhinia variegata stem bark extract. The fine powders of roots of Glycyrrhiza glabra and stem bark of Bauhinia variegata were subjected to methanolic, aquamethanolic and aqueous extraction and recovery of the extracts was 15.5, 19 and 10.24%, respectively, in case of Glycyrrhiza glabra and 30, 9.65 and 16.66 %, respectively for Bauhinia variegata. ABTS quenching activity (expressed as IC50 values) was 0.114 mg ml-1, 0.193 mg ml-1 and 0.709 mg ml-1 for methanolic, aqua-methanolic and aqueous extracts of Glycyrrhiza glabra, respectively whereas it was 0.276 mg ml-1, 0.302 mg ml-1 and 0.757 mg ml-1 for Bauhinia variegata extracts, respectively. DPPH radical quenching activity (expressed as IC50 values) was 0.159 mg ml-1, 0.571 mg ml-1 and 0.552 mg ml-1 for methanolic, aqua-methanolic and aqueous extracts of Glycyrrhiza glabra , respectively whereas it was 0.258 mg ml-1, 0.395 mg ml-1 and 0.410 mg ml-1 for Bauhinia variegata extracts, respectively. These results suggested that methanolic extract had better quenching activity for reactive radicals. The in vivo antioxidant activity was studied in rats (administred extract of both plants at dose of 100 mg kg-1 b.w. p.o. for 30 days) by evaluating lipid peroxidation, reduced glutathione and catalase in liver, kidney and erythrocytes of male Wistar rats (130-190g). Potassium dichromate at dose rate of 30mg kg-1 b.w. orally for 30days was used to induce oxidative stress in rats. The studies revealed a significant (P<0.05) decrease in lipid peroxidation in liver, kidney and erythrocytes and a significant (P<0.05) increase in reduced glutathione level in the liver and kidney of Bauhinia variegata and Glycyrrhiza glabra treated rats in comparison to oxidative stress induced rats. Further immunomodulatory studies were conducted in the rats immunocompromised with dexamethasone (5mg kg-1 b.w. interaperitoneally for 5 days). Dexamethasone caused immunosuppression characterized by significant lymphocytopenia.The studies further revealed a significant increase in footpad thickness of Glycyrrhiza glabra treated rats at 24 and 48 hours of challenge with 10 % SRBCs administered interadermally on sub planter region of footpad. There was a significant increase in the absolute lymphocyte count and haemagglutination titre in Glycyrrhiza glabra treated rats. On differential leucocyte count a significant lymphocytopenia observed in deaxamethasone treated rats. The histopathological evaluation of lymphoid tissue exhibited the enhanced lymphocyte proliferation in lymphoid follicles in Glycyrrhiza glabra and Bauhinia variegata treated rats. There was significant (P<0.05) increase in CD4+ receptor cell count in lymphocytes in Glycyrrhiza glabra treated rats in comparison to rats immunocompromised with dexamethasone. Further Glycyrrhhiza glabra had significantly (P<0.05) higher CD4+ cell count as compared to Bauhinia variegata treated rats and CD4+ cell count was comparable to placebo treated animals.The present studies revealed that Glycyrrhiza glabra has got a better antioxidant as well as immunomodulatory activity as compared to Bauhinia variegata.ThesisItem Open Access STUDIES ON PROTECTIVE EFFECTS OF SEABUCKTHORN (HIPPOPHAE RHAMNOIDES) OIL IN POULTRY AFLATOXICOSIS(CSKHPKV, Palampur, 2014-04) Pathak, Gyan Prakash; Varshneya, C.The present investigation was undertaken to study the protective actions of seabuckthorn (SBT) seed oil and glucomannan (GM) in aflatoxin (AF) treated broiler chicks. A total of 210, day old broiler chicks were used for studying the growth performance, biochemical parameter and antioxidant status for the period of 4 weeks. The seabuckthorn seeds oil was collected, SBT oil was fed orally to the broiler chicks @ 0.5 ml/kg bwt. and 1.0 ml/kg bwt. GM was mixed with the feed @ 1g/ kg whereas, AF was added @ 400ppb. The AF treatment significantly (P<0.05) reduced weight gain and increased FCR. The SBT oil as well as GM significantly (P<0.05) improved the growth performance. AF treatment produced a significant (P<0.05) decrease in total serum proteins, albumin, globulin, cholesterol, triglycerides and HDL and significant (P<0.05) increase in AST, ALT, ALP, LDH and total serum billirubin. However, SBT oil as well as GM significantly (P<0.05) restored these biochemical parameters to control levels. In aflatoxin treated groups the malondialdehyde (MDA) level of liver, kidney and blood were significantly (P<0.05) increased whereas GSH, Catalase, SOD and GPx levels were significantly (P<0.05) decreased, while there was no significant (P>0.05) change in these parameters were found following only SBT oil administration. The combined dietary supplementation of SBToil+AF and GM+AF produced significant (P<0.05) decrease in lipid peroxidation and an increase GSH, catalase, SOD and GPx level of liver, kidney and blood respectively than that of AF treated groups. The histopathological lesions in the liver were severe in AF treated group and were least severe in SBToil+AF followed by GM+AF groups and there were no lesions in only SBT oil alone treated groups. On the basis of growth performance, biochemical parameters, antioxidant activity and histology of the liver tissues, it is be concluded that oral supplementation of seabuckthorn oil @ 0.5 ml/kg bwt. has provides protection against aflatoxicosis in broilers.ThesisItem Open Access Effect of piperine on pharmaokinetics of gatifloxacin in dogs(CSKHPKV, Palampur, 2011) Sheria, Shalu; Varshneya, C.ThesisItem Open Access Studies on utilization of Seabuckthorn leaves for protection of Aflatoxicosis in poultry(CSKHPKV, Palampur, 2011) Thakur, Ashutosh; Varshneya, C.ThesisItem Open Access Study of ochra toxin induced nephropathy and its protection by seabuckthorn(Hippophae rhamnoides) Leaves in Japanese quail(CSKHPKV, Palampur, 2011) Jain, Aasheesh; Varshneya, C.ThesisItem Open Access Studies on antioxidant and immunomodulatory actions of seabuckthorn pomace in poultry(CSKHPKV, Palampur, 2010) Sharma,Ankita; Varshneya, C.