Dantrolene e células-tronco mensenquimais no tratamento do trauma medular agudo em ratos Wistar

Exportado OPUSMade available in DSpace on 2019-08-12T08:57:55Z (GMT). No. of bitstreams: 1 tese_final.pdf: 2028793 bytes, checksum: 1a520727c0db3da67305e4a459fffdcb (MD5) Previous issue date: 28Objetivou-se avaliar o efeito do dantrolene (DAN) e das células-tronco mesenquimais (CTM) no trauma medular agudo (TMA). Foram utilizados 60 ratos Wistar, machos com três meses de idade, divididos nos grupos CTM, DAN + CTM, DAN, controle positivo (CP) (trauma e placebo) e controle negativo (CN) (sem trauma e placebo). Realizou-se laminectomia de T12 em todos os grupos, seguida de TMA contusivo compressivo, com exceção do grupo CN. Uma hora depois, os grupos DAN + CTM e DAN receberam 10mg/kg de DAN em dose única, por via intraperitoneal e, após sete dias, os grupos CTM e DAN + CTM receberam 1x106 células, por via intravenosa. Testes comportamentais foram realizados para avaliar a recuperação funcional, por meio da escala de BBB e pelo método de escala descritiva adaptada, durante 30 dias. Para avaliação da medula espinhal foram realizadas histologia em H.E., imunoistoquímica com anti-NeuN e RT-PCR em tempo real para avaliar a expressão gênica dos fatores neurotróficos (BDNF e NT-3) e das proteínas antiapoptóticas (Bcl-2 e Bcl-xl) e pró-apoptóticas (caspase-3, caspase-9 e Bax), nos segmentos craniais e caudais ao epicentro da lesão. Os animais traumatizados apresentaram paraplegia grave e retenção urinária com cistite hemorrágica. Houve recuperação funcional significativa nos grupos tratados com CTM, DAN ou associação (p<0,05) em comparação ao grupo CP. Histologicamente, observou-se lesão espinhal grave, caracterizada por degeneração axonal e perda neuronal, estendendo-se craniocaudalmente ao epicentro. Todos os grupos apresentaram maior número médio de neurônios NeuN-positivos que os do CP (p<0,05). O TMA resultou em diminuição da expressão de BDNF, NT-3 e Bcl-xl, e aumento da expressão de caspase-3 e Bax em relação às medulas espinhais de animais do CN. Após o tratamento, observou-se aumento significativo da expressão de BDNF nos grupos CTM, CTM + DAN e DAN em relação ao grupo CP (p<0,05). Já a expressão da NT-3, mostrou-se maior nos grupos CTM + DAN e DAN nos segmentos craniais (p<0,05), e DAN nos caudais (p<0,01). A expressão de Bax foi significativamente menor nos grupos CTM + DAN (p<0,05) e DAN (p<0,001). A expressão das caspases 3 e 9 foi significativamente menor nos grupos DAN, CTM e CTM + DAN (p<0,05). Conclui-se que o DAN, as CTM ou associação de ambos, para tratamento de TMA em ratos, são protocolos seguros, promovem recuperação funcional e efeitos neuroprotetores, neurotróficos e antiapoptóticos..This study aimed to evaluate the effects of dantrolene (DAN) and mesenchymal stem cells (MSCs) in acute spinal cord injury (SCI). Sixty three-months-old male Wistar rats were divided into groups MSCs, MSCs + DAN, DAN, positive control (PC) (trauma and placebo) and negative control (NC) (no trauma and placebo). Laminectomy was performed at T12 level in all animals, followed by a weight-drop model of SCI, except for the NC group. An hour later, the MSCs + DAN and DAN groups received 10mg/kg of DAN in a single dose, intraperitoneally, and after seven days, the MSCs and MSCs + DAN groups received 1x106 cells intravenously. Functional recovery was assessed through BBB and adapted descriptive scale methods for 30 days. Spinal cord was evaluated with H.E., immunoistochemistry with anti-NeuN, and real time RTPCR to assess gene expression of neurotrophic factors (BDNF and NT-3), antiapoptotic (Bcl-2 and Bcl-xl) and pro-apoptotic proteins (caspase-3, caspase-9 and Bax), in the cranial and caudal segments from the lesion epicenter. Traumatized animals showed severe paraplegia and urinary retention with hemorrhagic cystitis. There was a significant recovery in groups MSCs, DAN and MSCs + DAN (p<0.05) compared to PC. Histologically, there was no systemic changes. It was observed severe spinal injury, characterized by axonal degeneration and neuronal loss, which extended cranio-caudally from epicenter. All groups showed average number of NeuN-positive neurons significantly higher when compared to PC (p<0.05). The SCI resulted in reduced BDNF, NT-3 and Bcl-xl gene expression, and increased Bax and caspase-3 expression. After treatments, there was a significant increase in BDNF expression in MSCs, MSCs + DAN and DAN groups compared to PC group (p<0.05). The NT-3 expression was higher in MSCs + DAN and DAN groups in cranial segments (p<0.05), and DAN group in the caudal ones (p<0.01). The Bax expression was significantly lower in the MSCs + DAN (p<0.05) and DAN (p<0.001). The caspase-3 and 9 expression were significantly lower in DAN, MSCs and MSCs + DAN groups (p<0.05). It was concluded that DAN, MSCs or the combination of both for the treatment of SCI in rats promote functional recovery and neuroprotection through neurotrophic and antiapoptotic effects

Epizootiology of canine distemper in naturally infected dogs in Goiânia, Brazil

ABSTRACT: Although the epizootiological profile of canine distemper in Goiânia is unknown, there is clinical evidence for a high incidence of canine distemper virus (CDV) infection among dogs. Therefore, this study determined the epizootiological characteristics of canine distemper in naturally infected dogs. Data of 46 dogs that tested positive for the CDV based on immunochromatography or reverse transcription-polymerase chain reaction were collected. Data on the sex, breed, age, and vaccination status were obtained from these dogs, and extraneural and neural sign analyses were performed. Although, the infected dogs belonged to both sexes, different breeds, and different age groups, a greater proportion of cases were seen in adults (1-6 years), undefined breeds, and unvaccinated dogs. Among the CDV-positive dogs, 10.87% had been vaccinated. In addition, 4.35% showed neural signs, 8.69% showed extraneural signs, and 86.96% showed both. High lethality was observed, with viral antigen and/or DNA detected in 82.61% dead dogs. Only 8.70% of the total CDV-infected dogs remained alive at the time of their assessment

Avaliação da técnica de guilhotina modificada para lobectomia hepática completa em gatos

O fígado, o maior tecido glandular do organismo, pode ser acometido por afecções localizadas, como neoplasias, abscessos e torção do lobo, onde é indicada a hepatectomia parcial ou a lobectomia. Nesse experimento, avaliou-se a aplicabilidade e segurança de uma forma modificada da técnica de sutura em guilhotina para a lobectomia hepática completa em gatos. Foram utilizados 13 gatos adultos jovens, sem raça definida, machos e clinicamente saudáveis. Empregou-se a técnica de sutura em guilhotina de forma modificada na base do lobo hepático lateral esquerdo, promovendo a hemostasia do órgão por meio de esmagamento dos tecidos. No período pós-operatório, foram avaliados diversos parâmetros clínicos por 10 dias consecutivos e aos 11 dias após a cirurgia realizou-se avaliação ultrassonográfica. Neste período, os parâmetros clínicos avaliados permaneceram dentro dos limites fisiológicos, demonstrando ausência de complicações importantes decorrentes da cirurgia. Na modificação da técnica de guilhotina utilizada, dividiu-se a base do lobo hepático em três segmentos, permitindo que a sutura promovesse um adequado esmagamento do tecido hepático e ligadura satisfatória dos vasos sanguíneos e ductos biliares, não sendo observada hemorragia ou ascite nos exames clínicos e ultrassonográficos. Aos 11 dias após a lobectomia, os fígados apresentavam-se ultrassonograficamente normais com pequena área de hiperecogenicidade no local da ligadura, sugestiva de tecido cicatricial. A técnica de sutura em guilhotina modificada foi considerada segura e com mínima morbidade para a lobectomia hepática em gatos

Estudo comparativo entre três técnicas abertas de orquiectomia em gatos

Background:  :  :  : Surgical sterilization of male cats is one of the most commonly procedures in the small animals practice, used as a control method against the overpopulation of domestic cats. Orchiectomy also aims to prevent or treat hormone-mediated diseases, disorders of the reproductive organs, such as testicular cancer, orchitis, prostatic diseases, trauma or abscesses. In addition, it may be used to control undesirable changes in behavior in adult cats such as the case of territorial marking and aggression. Several surgical techniques are described for orchiectomy in cats using different methods of hemostasis and/or ligation of the spermatic cord. There is a paucity of scientific studies and comparative data that supports one technique over the other, so this experiment was conducted to compare the three major open surgical techniques of ligation of the spermatic vessels to perform orchiectomy in cats. Materials, Methods & Results: Thirty-nine young entire adult male cats, with an average weight of 2.7 kg, were divided into three groups. In group I, animals underwent orchiectomy using the suture technique to ligation of the spermatic cord. In group II, the spermatic cord was used to perform an “eight-knot” on itself. In group III, the vas deferens and spermatic cord were used together to make “square-knots”. The outcome of surgical procedures was assessed by clinical examination, incidence of complications trans and postoperatively, duration of the procedure and ease of implementation. None of the animals had immediate or delayed bleeding after hemostasis. During surgery, there was no significant difference in procedure duration among the three different techniques. Postoperatively, the overall clinical evaluation was normal and there were no changes in behavior in any animal. It was noted a statistically significant difference in respiratory rate among groups, with the GI animals showing high frequency at the postoperative period when compared to animals from GII and GIII. There were no major complications in any of the techniques used and the parameters showed no significant difference among the groups. Discussion: The three most widely used techniques to perform orchiectomy in cats were evaluated in this study. Even though the respiratory rate has been shown to be statistically different among the groups, it was not a significant factor, because all other parameters were normal. These clinical data reflect the absence of major complications of surgery such as infection, pain, hypotension, hypovolemia or shock. In the current study, only adult cats were used and the structures of the spermatic cord showed strong and efficient ligation of testicular vessels with the techniques used. It was clinically observed that there was more edema in the group whose ligature employed was the suture technique. The catgut acts as a foreign body and interferes with the inflammatory reaction, the time of absorption and wound healing. The “eight-knot” technique is safe and effective, however, requires greater skill of the surgeon when compared to other techniques evaluated. The technique of “square-knot” resulted in less tissue manipulation without damaging the spermatic vessels, demonstrated by the absence of bleeding and minimal inflammatory reaction. This technique was effective, without complications postoperatively and easy to perform. These data suggest that the three techniques are considered easy and efficacious to perform orchiectomy in cats, but the technique of “square-knot” performed seems to be easier

Different leaf extracts from Brunfelsia uniflora in mice

ABSTRACT: In Brazil, at least 16 poisonous plant species can affect the central nervous system of livestock. Recently in the state of Piauí, Northeastern Brazil, Brunfelsia uniflora was reported as a cause of nervous signs in donkeys, cattle, and small ruminants. In order to assess the toxicity of B. uniflora extracts, 20 Swiss mice were distributed into four groups (n=5) receiving by gavage alkaloids, flavonoids, saponins, or saline (control group). After administration of the extracts in a single dose (5g kg-1), all mice were observed daily for clinical signs. Mice that received the extracts showed moderate to severe clinical signs, including piloerection, vocalization, and seizures. All mice dosed with saponins died between 10 and 20min after administration. Serum biochemical evaluation of animals that received saponins revealed slight increases in total protein levels and decreased magnesium and chlorite levels. In conclusion, saponins of B. uniflora leaves induced acute toxic neurological effects and death in mice