Theses

Browse

Search Results

Now showing 1 - 9 of 18

Open Access
Electroencephalographic and electrocardiographic studies on propofol anaesthesia in sheep
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2011-06) Raj Kumari; Das, Arup Kumar
In this study electroencephalographic and electrocardiographic changes during surgical interventions {viz., placement of central venous catheters (Group-B), docking/ tail amputation (Group-C), bilateral orchiectomy (Group-D) and no surgical stimuli as control (Group-A)} with propofol anaesthesia were assessed in twenty-four (n=24) apparently healthy male sheep. For surgical operations each sheep of this study was premedicated with glycopyrrolate (@ 0.02 mg/kg body weight, IM) and, five minutes after, anaesthesia was induced with propofol (@ 4.00 mg/kg body weight, IV). The parameters in this study included clinical, cardiopulmonary, haematobiochemical and electrocardiographic parameters. Similarly, qualitative electroencephalographic evaluation was done through spectral analysis to generate total power, SEF-90, MF and PPF over 0-30 frequency band width while relative power of different bandwidths viz., , , and was assessed. Rectal temperature and respiration rate has not shown any significant variability in any of the groups during the entire course of experiment. Haemoglobin concentration reduced and this extent was higher in control group than central venous catheterization, docking and castration groups. PCV reduced in all the groups. TEC depicted reduction in control group; however, this fall was not apparent in other groups. Blood glucose decreased initially followed by an increase in three groups after propofol induction while the castration group exhibited a non-significant increase from very beginning. Total serum protein declined in all the groups. A sudden increase in serum triglyceride (TG) was observed at one minute post-propofol induction in all the groups, which later on declined. Propofol has not been able to induce any significant changes in P-wave, QRS wave and T-wave amplitude as exhibited in control group. However, P-wave amplitude increased significantly in animals performed with central venous catheterization. The T-wave amplitude increased nonsignificantly in CVC group while decrease in docking and castration groups. P-wave and T-wave duration has not exhibited any significant difference throughout the course of experiment. However, QRS wave duration decreased in control group and no significant changes were observed in surgical groups. PR segment length reduced in castration group only and none of the other groups exhibited any significant changes. ST segment length exhibited significant increase in control group, non-significant increase in CVC while exhibiting significant fall in castration and docking groups. RR interval exhibited reduction in all the four groups, non-significantly in control while significantly in the three surgical groups. Heart Rate increased in all the four groups. EEG analysis showed an increase in total power, median frequency, theta frequency, peak power in all group, and after a transient increase reduction in spectral edge frequency and alpha frequency. However delta frequency showed an increase after a transient decrease and a marked decrease in beta frequency after propofol injection. It was concluded that propofol to some extent possess analgesic effect and may be useful in husbandry practices/ day case surgery. However, evaluation of analgesic effects of propofol needs further investigation.
Open Access
Studies on regenerative potential of hepatocytic mesenchymal cells in liver degeneration and delineation of their gene expression in canine
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-09) Saini, Rashmi; Jadon, N.S.
Twenty four dogs of either sex naturally suffering from hepatic dysfunction were used in this study. The animals were randomly divided into four groups administered with different drug or drugs combination viz. control (Group A); pepsid-C @ 1ml /10 kg body weight, IM alternatively for fifteen days (Group B); hepatocytic stem cells @ 5×107 cells, intraperitonially animal thrice at an interval of 5 days (Group C) and hepatocytic stem cells @ 5× 107 cells, intraperitonially animal thrice at an interval of 5 days + pepsid-C @ 1ml /10 kg body weight, IM alternatively for fifteen days (Group D). The clinical signs, haematological (Hb, TEC, TLC, DLC, PCV, BT and CT), biochemical (serum T.P, serum albumin, serum total bilirubin, serum glucose, serum urea nitrogen, serum creatinine and serum AST, serum ALT and serum GGT) parameters were studied at 0, 3rd, 6th, 10th, 15th and 20th day by using standard protocol. Radiographic and ultrasonographic changes were studied at 0 and 20th day following treatment. Gene expression of CD44 gene was evaluated via real time PCR. On the basis of parameter observed in this study, it was concluded that the effectiveness of treatment at 15th and 20th day interval was more in the animals treated with combination of hepatocytic stem cells + pepsid-C (Group D) and hepatocytic stem cells (Group C) alone as compared to pepsid-C alone (Group B), as indicated by haematological, biochemical, radiographic, ultrasonographic examination and gene expression of CD44 studies. Hepatocytic stem cells and pepsid-C are safe drugs used in the present study as there was no adverse biochemical and hematological changes during entire period of study. However, the variable changes observed during these periods were purely temporarily and were within normal physiological limits. Ultrasonographic study at day 0 interval of all the animals showed various degree of increased echogenicity, hepatopathy, changes in hepatic symmetric or asymmetric, cholecystitis, acute hepatitis and chronic hepatitis. Variable regenerative changes were seen in rest of the three groups B, C and D at 20th day following the administered of pepsid-C alone, hepatocytic stem cells alone and combination of hepatocytic stem cells + pepsid-C respectively. Abdominal radiograph of all the animals of group A, B, C and D at day 0 interval revealed the morphologic abnormalities in size, shape, position, and radio density of the liver. Gene expression of CD44 associated with regeneration was evident in animals where liver derived hepatocytes were administered and not evident in other group (A and B) while GAPDH, being a housekeeping gene was evident in all groups. Hepatocytic stem cells are necessary for the creation of new cells for growth, development and regeneration. These cells are responsible for the development of new specific cells and also have potential to differentiate into different cell types but are mostly bipotent and are specific for the type of tissue they belong to. These hepatocytic stem cells have better effect on liver function. Thus the combination of hepatocytic stem cells and pepsid-C may be used safely by field veterinarian for the treatment of hepatic dysfunction in canine.
Open Access
Apoptic effects of vincristine sulphate and cisplatin scaffolds on HeLa cell line and their clinical efficacy on canine transmissible venereal tumors
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-06) Arun Kumar; Jadon, N.S.
The present study was undertaken to evaluate the effects of vincristine sulphate, cisplatin and their scaffolds on canine transmissible venereal tumour in twenty four sexually mature adult dogs affected with naturally occurring canine transmissible venereal tumour (CTVT). The animals were randomly divided into four groups (n=6) and subjected to administration of different oncolytic drugs and drugs scaffolds. The animals of group A were administered vincristine sulphate @ 0.025 mg/kg intravenously once in a week for four consecutive weeks and animals of group B were administered cisplatin @ 2.14 mg/kg intravenously and repeated after 21 days. The animals of group C and D were subjected to the administration of scaffolds of vincristine sulphate @ 0.025 mg/kg intravenously once in a week for four consecutive weeks and scaffolds of cisplatin @ 2.14mg/kg intravenously and repeated after 21 days respectively. Preparation and characterization of hydrogel scaffolds were consisting of FTIR measurements, ultrastructure studies and electrochemical analysis. The oncolytic potential of the different chemotherapeutic agent (vincristine sulphate, cisplatin and their scaffolds) was evaluated on the basis of physical and cytological parameters, histopathological studies, haemato-biochemical parameters(Hb, PCV, TLC, DLC and platelets, total protein, glucose, BUN, creatinine, ALT, ASTand GGT) and apoptotic effect on HeLa cell line. Genomic DNA from HeLa cells was isolated and subjected to electrophoresis in agarose gel (1.8%) and 1kb DNA ladder. DNA fragments were visualized under a UV trans-illuminator and compared with a standard marker. Lane C showed no fragmented DNA, however, 20 μg/ml vincristine and 20 μg/ml cisplatin showed fragmented DNA in the form of ladder 1 and 3 after 24 h. Vincristine scaffolds and cisplatin scaffolds showed mild DNA fragment in lane 2 and 4. On the basis of parameter observed in this study, it is concluded that the early and best regression of the CTVT was observed in the animals treated with vincristine scaffolds. Cisplatin regressed the CTVT masses upto some extent; however, cisplatin scaffolds are moderately effective when it is used in appropriate dose. Vincristine alone is effective drug for the treatment of CTVT even in metastatic conditions, however the vincristine scaffolds are more effective as it has early regression of tumour as compare to vincristine alone. This may be due to decreasing the side effects caused in healthy cells. These vincristine scaffolds may be used safely by field veterinarian for the treatment of TVT in canines.
Open Access
Isolation and propagation of muscle-derived mesenchymal cells and assessment of their regenerative potential in skeletal muscle injuries in mice
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Godiyal, Akanksha; Kandpal, Manjul
The present study was conducted on 24 clinically healthy Swiss albino mice (8-10 weeks old) of either sex, weighing 25-35 gm. The animals were divided into three groups viz. group A, B and C, having 8 animals in each. In all the groups, the gastrocnemius muscle was injured by surgical resection of the muscle. After creating muscle injury, PBS was injected in the mice of group A (control group) on day 1 and 5. In the mice of group B, MDSCs were injected in the injured muscle on day 1 and 5. While in the mice of group C, MDSCs were injected in the injured muscle on day 5 and 10. Muscle-derived mesenchymal cells were isolated by modified preplate method and cultured in proliferation media and incubated at 37 °C in a humidified 5% CO2 incubator. When the cells reached 70-80 % confluency, they were harvested using 0.25% trypsin-EDTA and cells were implanted at the site of injury in animals of group B and C. The regenerative potential of MDSCs for repair of the injured muscle was assessed by the evaluation of physical parameters, physiological parameters, wound condition, biochemical parameters, histopathological studies and free wire hanging test at different time intervals. Also, gene expression profiling of mVEGF was done for the evaluation of muscle regeneration via angiogenesis on day 7 and 15 after creation of muscle injury wound. On the basis of parameters observed in the present study, early and better healing of injured muscle was revealed in the mice of group B. There was early reduction in swelling, exudation, warmth and pain at the repaired site in the mice of group B. The level of enzymes CK, LDH, AST and AST was decreased significantly in the mice of group B. Also, the score of free wire hanging test was highest in the mice of group B as compared to the mice of group A and C indicating complete regeneration of the injured muscle. Histopathological observations revealed the presence of well organized, polygonal shaped regenerated myofibers having peripherally arranged nucleus in the mice of group B. Gene expression analysis showed upregulation of the gene mVEGF in group B as compared to control group. Results concluded that muscle-derived mesenchymal cells can be isolated by modified preplate method, cultured in-vitro and could be used without any scaffold for regeneration of injured skeletal muscle. Therefore, the current study confirmed that muscle-derived mesenchymal cells may be used for early, better and complete muscle regeneration in clinical cases of muscle injuries.
Open Access
Studies on the chemotherapeutic effects of Docetaxel and gene expression during regression of mammary tumour in canine
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Upadhyay, Prachi; Jadon, N.S.
Sixteen adult dogs suffering from canine mammary tumours used in this study were divided randomly into two groups having equal number of dogs (Group I and Group II). Patients of group I were subjected to administration of docetaxel (@30mg/m2 weekly four consecutive cycles) and patients of Group II were subjected to surgical excision of tumoral growth followed by chemotherapy with docetaxel (@30mg/m2 weekly four consecutive cycles). The therapeutic efficacy was determined by observing various parameters clinical (physical appearance and gross regression), radiological assessment, ultrasonographic screening, haematological (haemoglobin, total erythrocyte count, total leucocyte count and differential leucocyte count), biochemical (aspartate amino transferase, alanine amino transferase, serum creatinine and serum urea nitrogen) histopathological and gene expression profiling of EGFR to assess the fold change in its expression as result of treatment modality via RT-PCR analysis. The rectal temperature, heart rate and respiration rate showed non-significant changes at various time intervals in both the groups. Haemoglobin levels decreased non-significantly, total erythrocyte count and platelets varied non-significantly in both the groups. Neutrophil count revealed significant decrease in both the groups, whereas significant increase in lymphocyte increased significantly in both the groups and significant increase was observed in group II as compared to group I. Biochemical study revealed significant increase in levels of AST, BUN and serum creatinine within group II further, significant increase was also observed in group II as compared to group I. istopathologically, five (31.25%) tumours appeared to be benign while eleven (68.75%) tumours were malignant. On ultrasonographical study, no difference in regularity of tumour shape and margins, echotexture and shadowing was observed among benign and malignant tumours, tumour echogenicity was homogenous in benign tumours and posterior enhancement was evident in malignant ones. Lateral and ventrodorsal radiographs manifested lung metastsis in 10 cases however, three cases out of eight in group I and one case out of eight in group II revealed slight lung clearence on day 28. Clinical response rate was responders 62.5% and non-responders 37.5% in group I. In group II, complete response with no reoccurrence was observed in five cases (62.5%) and three cases showed reoccurrence, these patients had a tendency towards better survival rates and prolongation of life as compared to group I. Gene expression profiling revealed that EGFR receptor was comparatively more down-regulated in patients of group II subjected to surgical resection of canine mammary tumours followed by chemotherapy as compared to patients of group I which were subjected to docetaxel therapy. On the basis of above mentioned parameters it was concluded that the combination therapy had better response rate, survival rates and more down-regulation of EGFR gene involved in tumour invasion and metastasis. Combination of surgery and chemotherapy with docetaxel may be used safely by field veterinarians under proper observation for the treatment of mammary tumours in canines.
Open Access
Anaesthetic evaluation of various combinations of acepromazine, butorphanol and propofol in dogs
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Rohit Prabhat; Kandpal, Manjul
Eighteen adult dogs (requiring various clinical procedures) used in this study were divided randomly into three groups (A, B and C). All the dogs were premedicated with injection atropine sulphate at the dose rate of 0.04mg/kg body weight I/M in all the groups. Thereafter, acepromazine at the dose rate of 0.05mg/kg body weight I/M, butorphanol at the dose rate of 0.4mg/kg I/M and combination of acepromazine (at the rate of 0.05mg/kg body weight I/M) and butorphanol (at the rate of 0.04mg/kg body weight I/M), were administered in the group-A, -B and -C, respectively. The general anaesthesia using propofol in the group-A, -B and -C were found to be, 6.13±2.50mg/kg body weight, 4.84±1.97 mg/kg body weight and 3.49±1.42mg/kg body weight, respectively on intravenous administration. Maintenance of propofol with repetitive administration of established dose was done on the basis of response. The effectiveness of anaesthesia was evaluated by observing various clinical (Induction time, duration of anaesthesia, muscle relaxation, recovery time, required doses of induction agent, physiological (rectal temperature, heart rate, respiration rate, blood pressure, SpO2, electrocardiography), haematological (haemoglobin, total erythrocyte count, total leucocyte count, differential leucocyte count, packed cell volume, and biochemical (serum glucose, total protein, serum urea nitrogen, serum creatinine, serum bilirubin, alanine amino transferase and aspartate amino transferase) parameters before and after administration of anaesthesia. Group-C showed a quicker induction, better analgesia and muscle relaxation as compared to the other groups. Heart rate and respiratory rate increased significantly in group-A after anaesthesia. In group-C mean arterial blood pressure was increased as compared to other groups. In all the groups haematological parameters maintained their values within the normal range significantly. Serum glucose and total protein values altered significantly within their normal range in all the groups but total protein was somewhat higher in group-A. The creatinine and blood urea nitrogen vary significantly within the normal range. AST, ALT and total bilirubin level was significantly reduced transiently and then maintained within the normal range in all the groups. On the basis of clinicophysiological and haematobiochemical observation, all the anaesthetic combination were found suitable and effective in canine general surgery. Propofol with its smooth induction, short duration of action, early recovery and good compatibility with acepromazine and butorphanol, either alone or in combination, was found to be an effective general anaesthetic. Being a neuroleptanalgesia, acepromazine and butorphanol with propofol (group-C) was found suitable for surgeries demanding pain salvage for long duration.
Open Access
Studies on biocompatibility and biodegradability of magnesium based orthopaedic bone implants in avian model
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-06) Abhishek, M.S.; Das, Arup Kumar
The present study was conducted in 18 adult Uttara-fowls of 8-10 months of either sex, divided into three groups with six birds each, for intramedullary insertion of Mg and Mg-apatite orthopaedic bone spacers with the objective of finding their biodegradability and biocompatibility for the period of 180 days. In first, second and third group, plain Magnesium (Mg), Mg with five per cent Hydroxyapatite (HA) and Mg with 15 per cent HA containing implants, were surgically inserted in intramedullary space of humerus, respectively. The anaesthetic regimen with atropine sulphate premedication and induction of ketamine anaesthesia was pursued. For radiography medio-lateral and leading edge (Hanging-drop positioning technique) views for humerus of the birds were taken on scheduled intervals (immediate postoperative, 1st week, 2nd week, 3rd week, 4th week, 6th week, 8th week,10th week, 12th week, 15th week, 18th week, 21st week and 24th week) and clinical parameters like heart rate, cloacal temperature, surgical duration, flight test, wing dropping test, histological evaluation and serum magnesium, calcium and phosphorous estimation were assessed to evaluate biodegradability and biocompatibility of implant material. Similar postoperative treatment and care were given to every bird during surgical convalescence. At the end of the study all the birds were euthanized with intravenous injection of thiopentone sodium as recommended by The Animal Welfare Board of India. Initiation of biodegradation was discernible in radiographs in birds of Group-I on 1st-2nd week (11th day); in Group-II on 3rd week (17th day); and Group-III on 1st week (6th day). In this study the biodegradable magnesium based orthopaedic bone implants were evaluated as the best choice for the avian fracture management. From this study following conclusions can be made (i) all Mg based metallic implants are biocompatible and biodegradable (ii) concentration of HA relates directly in the biodegradation of the Mg-HA matrix implant.
Open Access
Isolation, propagation and characterization of bone marrow derived mesenchymal stem cells and their differentiation into neurocytes in dogs
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-07) Verma, Rakesh Kumar; Kandpal, Manjul
The adult stem cell therapy is a blooming area of clinical research and relevance. The therapeutic benefits of mesenchymal stem cells in veterinary regenerative medicine are numerous as they are in human medicine. The mesenchymal stem cells in canines can be potentially used for cell based therapy to regenerate damaged or lost neuronal cells. The leitmotif of this study was differentiating cBM-MSCs into neurocytes in vitro. A healthy nondescript male dog of 3 years of age and 24kg weight was used for this study. The bone marrow aspirate was aseptically collected from ileac crest of the dog after administration of general anaesthesia by 2.5% thiopentone sodium till effect and atropine sulphate and diazepam as pre-anaesthetics. The bone marrow aspirate was processed in laboratory within 4 hours of its collection. Mesenchymal stem cells were isolated by density gradient centrifugation technique and cells were seeded in T-25 tissue culture flasks. After attainment of 80-90% confluence, first passage was performed by using Trypin-EDTA to expand the cell population. Cells were passaged till 4 passages and then grown in neurogenic differentiation media for 21 days in CO2 incubator at 37°C and 5% CO2. The morphology of cells was transitioned from spindle elongated shape to ring or net like shape. The cells were characterized at regular intervals by staining with the Nissl body stain after 21 days with positive result which coloured the nuclei blue-violet. The RT-PCR for the genes NSE and NFM revealed bands on days 21 confirming the differentiation process. The sample on gelatin scaffold and on pellet when subjected to scanning electron microscopy revealed net shape cells with cell aggregates. On the basis of this study it was concluded that MSCs possess potential to transdifferentiate to neurocytes when cultured in neurogenic media in vitro and the identification techniques are successful. The meticulous exploitation of MSCs in tissue engineering and regenerative therapy in veterinary medicine is the need of the hour.
Open Access
Isolation, expansion and characterization of bone marrow derived mesenchymal stem cells and their differentiation into hepatocytes in canines
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-07) Shaista Khanum; Kandpal, Manjul
Mesenchymal stem cells (MSCs), as adult stem cells (ASCs) able to divide into a variety of different cells, are of utmost importance for stem cell research. Mesenchymal stem cells (MSC) isolated from bone marrow and differentiated into hepatocyte-like cells have increasingly gained attention for clinical cell therapy of liver diseases because of their high regenerative capacity. The main focus of current study was on canine bone marrow derived mesenchymal stem cells and their differentiation towards hepatic lineage in vitro. The bone marrow was aspirated from iliac crest of young non-descript male dog under xylazine and ketamine anesthesia. The mononuclear cells along with mesenchymal stem cells were harvested by density gradient centrifugation. The mesenchymal stem cells were isolated on the basis of their plastic adherence property. The cells were seeded in tissue culture flasks and allowed to grow in monolayer till the attainment of 80-90% of confluency. At this confluency passaging was done using trypsin-EDTA and cultured with two different densities of 16 cells/cm2 and 4000 cell/cm2 in T-25 flasks. The MSCs cultured with initial seeding density of 4000 cell/cm2 showed higher yield up to 10 passages with faster expansion and followed standard growth kinetic curve. Thus, the fourth passage of these MSCs were made to differentiate into hepatocytes in stepwise manner including induction, differentiation and maturation steps. The differentiation was done by using growth factors HGF, bFGF and nicotinamide for induction, HGF, ITS and Dexamethasone for differentiation and OSM, ITS and dexamethasone for maturation. The cells were cultured for 30 days in an incubator maintained at 5% CO2 and 37oC.The morphology of cells was observed at regular intervals and cells were characterized by PAS staining, ICG uptake, gene expression analysis and SEM. The cells demonstrated positive PAS staining by giving pink to purple red colour. The cells also affirmed the ICG uptake by manifestation of dark green colour nuclei. The RT-PCR analysis revealed the expressions of hepatocyte specific markers alfa fetoprotein (AFP), albumin (ALB), tyrosine aminotransferase (TAT) and alpha-1 antitrypsin (_1-AT) in differentiated cells. The cells when subjected to scanning electron microscopy also revealed hepatocyte like morphology. On the basis of this study it was concluded that canine bone marrow derived mesenchymal stem cells were expanded successfully up to 10 passages with optimum initial seeding density and possessed potential for differentiating into hepatocyte-like cells in vitro. The exploitation of this in vitro harvested hepatocyte like cells may prove an effective therapy for liver diseases.

Theses

Browse

Thesis Type

Type

Author

Date

Subject

Search Results