865 resultados para pre-clinical animal models
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Members of the subfamily Alphaherpesvirinae use the epithelium of the upper respiratory and/or genital tract as preferential sites for primary replication. However, bovine herpesvirus 5 (BoHV5) is neurotropic and neuroinvasive and responsible for meningoencephalitis in cattle and in animal models. A related virus, BoHV1 has also been occasionally implicated in natural cases of neurological infection and disease in cattle. The aim of the present study was to assess the in vitro effects of BoHV1 and BoHV5 replication in neuron-like cells. Overall, cytopathic effects, consisting of floating rounded cells, giant cells and monolayer lysis, induced by both viruses at 48 h postinfection (p.i.) resulted in a loss of cell viability and high virus titres (r = 0.978). The BoHV1 Cooper strain produced the lowest titres in neuron-like cells, although viral DNA was detected in infected cells during all experiments. Virus replication in infected cells was demonstrated by immunocytochemistry, flow cytometry and qPCR assays. BoHV antigens were better visualized at 48 h p.i. and flow cytometry analysis showed that SV56/90 and Los Angeles antigens were present at higher levels. In spite of the fact that BoHV titres dropped at 48 h p.i, viral DNA remained detectable until 120 h p.i. Sensitive TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) and annexin V assays were used to identify apoptosis. BoHV5 induced death in approximately 50 % of cells within 24 h p.i., similar to what has been observed for BoHV1 Los Angeles. Infection with the BoHV1 Cooper strain resulted in 26.37 % of cells being in the early stages of apoptosis; 63.69 % of infected cells were considered viable. Modulation of mitochondrial function, as measured by mitochondrial membrane depolarization, was synchronous with the virus replication cycle, cell viability and virus titres at 48 h p.i. Our results indicate that apoptosis plays an important role in preventing neuronal death and provides a bovine-derived in vitro system to study herpesvirus-neuron interactions.
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Introduction: The urgent need for studies using standardized protocols to evaluate the real biological effects of PRP has been emphasized by several authors. Objective: The purpose of this study was to standardize a methodology for autologous Platelet-Rich Plasma (PRP) preparation in rats. Material and methods: Twentyfour, 5 to 6-month-old, male rats, weighing 450 to 500 g were used. After general anesthesia, 3.15 ml of blood was collected from each animal, via cannulation of the jugular vein. A standardized technique of double centrifugation was used to prepare PRP. PRP samples and peripheral blood platelets were then manually counted using a Neubauer chamber. Student’s t-test was used to compare the differences between the number of platelets in peripheral blood and PRP samples (p < 0.05). In addition, PRP and peripheral blood smears were stained to see platelets’ morphology. Results: All surgical procedures were well tolerated by the animals and they were healthy during the entire experimental period. PRP samples showed higher significantly platelet concentrations than peripheral blood samples (2,677,583 and 683,680 respectively). Conclusion: Within the limits of this study, it can be concluded that the method used produced autologous PRP with appropriated platelet quantity and quality, in rats.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
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Pós-graduação em Odontologia Restauradora - ICT
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Objective: To present a model for research and training in multivisceral transplantation in pigs. Methods: Eight Large White pigs (four donors and four recipients) were operated. The multivisceral transplant with stomach, duodenum, pancreas, liver and intestine was performed similarly to transplantation in humans with a few differences, described below. Anastomoses were performed as follows: end-to-end from the supra-hepatic vena cava of the graft to the recipient juxta diaphragmatic vena cava; end-to-end from the infra-hepatic vena cava of the graft to the inferior (suprarenal) vena cava of the recipient; and endto-side patch of the aorta of the graft to the infrarenal aorta of the recipient plus digestive reconstruction. Results: The performance of the multivisceral transplantion was possible in all four animals. Reperfusions of the multivisceral graft led to a severe ischemia-reperfusion syndrome, despite flushing of the graft. The animals presented with hypotension and the need for high doses of vasoactive drugs, and all of them were sacrificed after discontinuing these drugs. Conclusion: Some alternatives to minimize the ischemia-reperfusion syndrome, such as the use of another vasoactive drug, use of a third pig merely for blood transfusion, presence of an anesthesia team in the operating room, and reduction of the graft, will be the next steps to enable experimental studies.
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Recent research seeking to elucidate the possible effects of different types of physical training on the morphological adaptations of skeletal muscle. Although it is relatively easy to study the effects of exercise training in humans, such research becomes limited due to the invasive nature of the biopsies and the risk inherent in the use of human subjects. Thus, the application of animal models of training has been considered an appropriate strategy for the study of muscular adaptations in response to exercise. Objective: This study used a rodent model to determine the possible effects of aerobic and strength training on the CSA of fibers of the plantaris muscle. Methods: 24 male Wistar rats (80 to 120 days, 250 to 400 g) were randomly divided into 3 groups: aerobic training (TA, n = 8), strength training (ST, n = 8) and control (CO, n = 8). The animals in groups TA and TF were subjected to 8 weeks of training, while the animals of group C remained without any stimulus from start to finish the training period. At the end of the experiment, the animals were sacrificed and right plantar muscles dissected and removed. For morphological and morphometric analysis of muscle fibers was performed staining was performed H.E. Results: There was no significant difference in initial body weight between experimental groups. After 8 weeks of training, the TA group showed a significant reduction in final body weight, compared to CO and TF groups. With respect to the CSA of fibers of the plantaris muscle, no significant difference between the groups CO and TA. On the other hand, the strength training promoted a significant increase in AST of the group TF in compared with the groups CO and TA. Conclusion: Strength training used in this study promoted an increase in CSA of fibers of the plantaris muscle. On the other hand, animals submitted to aerobic training showed no changes in the CSA of the fibers, however, there was reduction in PC animals. The data strongly suggest the use of animal model of strength training used in this study as an appropriate strategy for studying the hypertrophic response of skeletal muscle.
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Tuberculosis due to Mycobacterium bovis affects both captive and free-ranging Cervidae in the United States. Various animal models have been developed to study tuberculosis of both humans and animals. Generally, tuberculosis is transmitted by aerosol and oral routes. Models of aerosol exposure of large animals to M. bovis are uncommon. In order to develop a reliable method of aerosol exposure of white-tailed deer (Odocoileus virginianus) to M. bovis, 12 healthy white-tailed deer, aged 8–10 mo, were infected by aerosol exposure to 2x105 to 1x106 colony forming units (CFU) (high dose, n=4) of M. bovis or 6x102 to 1.6 x 103 CFU (low dose, n=8) of M. bovis. Tuberculous lesions were more widely disseminated in deer receiving the high dose, while lesions in deer receiving the low dose were more focused on the lungs and associated lymph nodes (tracheobronchial and mediastinal). Aerosol delivery of M. bovis to white-tailed deer results in a reliable manner of experimental infection that may be useful for studies of disease pathogenesis, immune response, mycobacterial shedding, and vaccine efficacy.
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The following study analyzed the attitudes held by pre-clinical medical students about the Medical College Admission Test or MCAT. One hundred and eighty first-year and second-year medical students at a public Midwestern medical university participated in this study. Participants completed the “Medical Students Attitudes toward the Medical College Admission Test” survey during their morning lectures near the end of their spring semester. A composite scale score of the Likert items of the survey was computed and the proportion of students with attitudes ranging from strongly agree to strongly disagree was calculated. For six of the twelve Likert items the largest proportion of participants disagreed with the statements about the MCAT and its use in the admission process and its applicability to their current medical education. Other questions included how participants prepared for the MCAT and if they completed each of the subsections were addressed as well. Future research could determine if attitudes between students accepted into medical school and those not accepted are drastically different. Advisor: Kurt F. Geisinger