Paulo Fantinato Neto

Fetal-Maternal Interactions in the Synepitheliochorial Placenta Using the eGFP Cloned Cattle Model

Background To investigate mechanisms of fetal-maternal cell interactions in the bovine placenta, we developed a model of transgenic enhanced Green Fluorescent Protein (t-eGFP) expressing bovine embryos produced by nuclear transfer (NT) to assess the distribution of fetal-derived products in the bovine placenta. In addition, we searched for male specific DNA in the blood of females carrying in vitro produced male embryos. Our hypothesis is that the bovine placenta is more permeable to fetal-derived products than described elsewhere. Methodology/Principal Findings Samples of placentomes, chorion, endometrium, maternal peripheral blood leukocytes and blood plasma were collected during early gestation and processed for nested-PCR for eGFP and testis-specific Y-encoded protein (TSPY), western blotting and immunohistochemistry for eGFP detection, as well as transmission electron microscopy to verify the level of interaction between maternal and fetal cells. TSPY and eGFP DNA were present in the blood of cows carrying male pregnancies at day 60 of pregnancy. Protein and mRNA of eGFP were observed in the trophoblast and uterine tissues. In the placentomes, the protein expression was weak in the syncytial regions, but intense in neighboring cells on both sides of the fetal-maternal interface. Ultrastructurally, our samples from t-eGFP expressing NT pregnancies showed to be normal, such as the presence of interdigitating structures between fetal and maternal cells. In addition, channels-like structures were present in the trophoblast cells. Conclusions/Significance Data suggested that there is a delivery of fetal contents to the maternal system on both systemic and local levels that involved nuclear acids and proteins. It not clear the mechanisms involved in the transfer of fetal-derived molecules to the maternal system. This delivery may occur through nonclassical protein secretion; throughout transtrophoblastic-like channels and/or by apoptotic processes previously described. In conclusion, the bovine synepitheliochorial placenta displays an intimate fetal-maternal interaction, similar to other placental types for instance human and mouse.

Respostas fisiológicas e comportamentais de vacas Holandesas mantidas em sistema adiabático evaporativo

This study aimed to evaluate the influence of cooling systems, ventilation and nebulization on the physiology and behavior of Holstein cows housed in free-stall during the summer of southeastern Brazil. 20 Holstein cows were subjected to two treatments with and without cooling system. Environmental parameters dry bulb temperature, relative humidity and black globe temperature were also recorded. Rectal temperature and respiratory rate were evaluated at 5h, 9h30min, 11h30min, 13h30min, 16h30min, 18h30min e 21h30min. The behavioral variables recorded were posture and activities from 5h to 21:30h. Statistics we done using the method of least squares means. Despite the statistical differences obtained for the physiological variables, they were not biologically effective and indicated that the animals were in thermal comfort. Animals that had ventilation and nebulization have eaten even during the hottest hours of the day. The cooling system is a strategy that allows greater thermal comfort to animals and therefore can optimize milk production by increasing the dietary intake.

Investigação clínica e comparação do esfregaço sanguíneo e PCR para diagnóstico de hemoparasitas em equinos de esporte e tração (carroceiros)

Na clinica medica de equinos, explora-se o perfil hematologico do animal, geralmente, com a finalidade de encontrar alteracoes que nao foram constatadas ao exame clinico. A pesquisa de hematozoarios em equinos, muitas vezes, apresenta resultados conflitantes entre o quadro clinico apresentado pelo animal e o resultado laboratorial, levantando a hipotese de que a tecnica de pesquisa de hematozoarios seja a responsavel por falhas diagnosticas. Este estudo visa comparar os valores obtidos em exames hematologicos de 15 equinos de esporte e 15 equinos de tracao (carroceiros), levando-se em consideracao diferencas como caracteristicas nutricionais, estado de higidez e tipo de atividade realizada, e comparar as diferentes tecnicas de pesquisa de hematozoarios, como esfregaco sanguineo e PCR. Verificou-se que apenas os equinos de tracao apresentaram valores medios de hemacias, hematocrito e hemoglobina abaixo do considerado fisiologico para a especie, embora 100% dos animais, de ambos os grupos experimentais, tenham sido considerados positivos para hemoparasitoses por PCR. Verifica-se a superioridade do metodo de pesquisa de hemoparasitas por PCR, em comparacao com esfregaco sanguineo, realizado por diferentes tecnicas, visto que apenas 33,3% dos animais foram considerados positivos para Theileria equi por esta tecnica, enquanto que o PCR revelou 100% de positividade, para Theileria equi, Babesia caballi e infeccao mista. Nenhum dos animais estudados foi diagnosticado com Anaplasma phagocytophilum (Ehrlichia equi) e Ehrlichia risticcii (Neoricketsia risticii). Verifica-se, entao, que muitos dos diagnosticos de ausencia de hemoparasitose por exame hematologico e ou esfregaco sanguineo sao erroneos, devido a baixa sensibilidade da tecnica e podem repercutir em falha no tratamento ou disseminacao dos hemoparasitos e das hemoparasitoses. Ressalta-se, entao, a importância de exames como o PCR na elaboracao de diagnostico definitivo.

Tepoxalin on renal function and liver enzymes in cats exposed to hypotension with isoflurane

This study aimed to evaluate the possible renal and hepatic toxicity of tepoxalin administered before or after isoflurane-induced hypotension, as well as for five consecutive days. Twelve healthy mixed-breed cats, adult males, weighing 4.0±0.8kg were allocated into two groups. They received 25mgkg-1 of tepoxalin orally, two hours before the anesthetic procedure (PRE) or after the procedure (POST) and daily for five days. Cats were anesthetized with isoflurane and the concentration was increased until mean arterial pressure reached 40-60mmHg and kept at this level for 60 minutes. During hypotension, the physiological variables were measured at time 0 and every 10 minutes until 60 minutes, and bleeding time was measured at time 0, 30 and 60 minutes. Blood samples were drawn for a hemogram and determination of concentrations of alanine aminotransferase, alkaline phosphatase, urea, creatinine and Na+ at baseline, 24 hours, 48 hours and 7 days post-hypotension. Urine was collected at baseline, 24 hours, 48 hours and 7 days post-hypotension for determination of concentrations of creatinine, gamma-glutamyltransferase, urine specific gravity, protein, albumin and Na+. During the anesthetic procedure there were no important variations in physiological variables and bleeding time. There were differences only in fractional excretion of Na+, which was elevated at 7 days of evaluation in PRE and in the urine protein/creatinine ratio in PRE, which was higher than in POST at 24 and 48 hours post-hypotension. We conclude that tepoxalin does not cause alterations in hepatic enzymes but can cause discrete renal injury, resulting in proteinuria, in cats subjected to 60min of hypotension.