A influência do óxido nítrico nas alterações patológicas tardias do músculo esquelético ocasionadas pela sepse induzida pelo método Cecal Slurry

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Biologia Celular e do Desenvolvimento, Florianópolis, 2022.A sepse é responsável por cerca de 20% de todas as mortes mundiais. Aproximadamente, 40% dos sobreviventes desenvolvem a síndrome pós-sepse, cujos principais problemas estão relacionados com disfunções do músculo esquelético. Essas disfunções impactam tanto a qualidade de vida como a sobrevivência a longo prazo desses indivíduos. O aumento do óxido nítrico (NO) durante a sepse parece ter relação com o desenvolvimento desse dano muscular. Estudos pré-clínicos de sepse utilizam como padrão-ouro o modelo CLP (Cecal Ligation and Puncture) em camundongos, porém, o método Cecal Slurry utilizado para indução de sepse emcamundongos C57BL/6 parece ser mais vantajoso, pela ausência da necessidade de realização de cirurgia e por não ter diferença em relação ao tamanho e quantidade do conteúdo cecal dos animais. Portanto, um dos nossos objetivos foi adaptar o método Cecal Slurry para camundongos Swiss, avaliando parâmetros comumente utilizados para caracterizar a sepse em roedores. Em um segundo objetivo, utilizamos o modelo Cecal Slurry para verificar a influência do NO na miopatia tardia observada no músculo esquelético causada pela sepse. Para este fim, induzimos a sepse pelo método previamente citado, e após 24 h iniciamos o tratamento dos animais com um inibidor não seletivo das óxido nítrico sintases (NOS; L-NAME) e um inibidor seletivo da isoforma neuronal (NOS-1) das NOS (7-NI), com ambas as drogas inibindo seletivamente ou não a produção de NO. Além disso, utilizamos um doador de NO, o nitroprussiato de sódio (SNP), com objetivo de fornecer exogenamente o NO nos animais. Após 10 dias da indução da sepse e tratamento com as drogas realizamos análises morfométricas e imuno-histoquímicas do músculo EDL (extensor digitorum longus). Nossos resultados mostram que o método foi padronizado com sucesso nos animais Swiss, por ter induzido um quadro típico de sepse. Os animais apresentaram hipotermia, redução de peso corporal, diminuição da ingestão alimentar, hipoglicemia, hipotensão e aumento de creatinina nos camundongos sépticos, estatisticamente diferentes dos animais controle. Esse quadro de sepse teve uma taxa de sobrevivência de 25%. As análises do músculo EDL apresentaram alterações patológicas após dez dias da indução da sepse, mais especificamente atrofia, edema e aumento do espaço intersticial entre as miofibras. A realização da imuno-histoquímica para a NOS-1 teve marcações no núcleo, sarcolema e no interior das miofibras. A quantificação da NOS-1 foi estatisticamente menor no grupo tratado com inibidor de NOS. Com isso, nosso trabalho mostra que a indução da sepse pelo método Cecal Slurry ocasionou em alterações patológicas tardias no músculo esquelético, e a inibição não seletiva de NOS parece ter sido o tratamento que mais apresentou dano muscular no EDL, pois sofreu a maior redução da enzima NOS-1 nesse órgão, e em outras miopatias a ausência de NOS-1 está relacionada com um dano muscular maior. Porém, são necessários mais estudos, como a avaliação funcional, para a confirmação desse evento.Abstract: Sepsis is responsible for about 20% of all deaths worldwide. Approximately 40% of survivors develop post-sepsis syndrome, whose main problems are related to skeletal muscle dysfunction. These dysfunctions impact both the quality of life and the long-term survival of these individuals. The increase in nitric oxide (NO) during sepsis seems to be related with the development of this muscle damage. Preclinical studies of sepsis use the CLP (Cecal Ligation and Puncture) model in mice as the gold standard, however, the Cecal Slurry method used to induce sepsis in C57BL/6 mice seems to be more advantageous, due to the absence of the need for surgery and because there is no difference according to the size and quantity of the animals' cecal content. Therefore, one of our objectives was to adapt the Cecal Slurry method for Swiss mice, evaluating parameters commonly used to characterize sepsis in rodents. In a second objective, we used the Cecal Slurry model to verify the influence of NO on the late myopathy observed in skeletal muscle caused by sepsis. For this purpose, we induced sepsis by the previously mentioned method, and after 24h we started the treatment of the animals with a non-selective nitric oxide synthases inhibitor (NOS; L-NAME) and a selective inhibitor of the neuronal isoform (NOS-1) of NOS (7- NI), with both drugs selectively or not selectively inhibiting NO production. In addition, we used a NO donor, sodium nitroprusside (SNP), with the aim of exogenously supplying NO to the animals. Ten days after sepsis induction and drug treatment, we performed morphometric and immunohistochemical analyzes of the EDL muscle (extensor digitorum longus). Our results show that the method was successfully standardized in Swiss animals, as it induced a typical condition of sepsis. The animals showed hypothermia, reduced body weight, decreased food intake, hypoglycemia, hypotension and increased creatinine in septic mice, statistically different from control animals. This sepsis condition had a 25% survival rate. Analysis of the EDL muscle showed pathological changes ten days after sepsis induction, more specifically atrophy, edema and increased interstitial space between myofibers. Immunohistochemistry for NOS-1 showed markings in the nucleus, sarcolemma and inside the myofibers. NOS-1 quantification was statistically lower expressed in the group treated with NOS inhibitor. Thus, our work shows that the induction of sepsis by the Cecal Slurry method caused late pathological changes in the skeletal muscle, and the non-selective inhibition of NOS seems to have been the treatment that most showed muscle damage in the EDL, as it suffered the greatest reduction in NOS-1 enzyme in that organ, and in other myopathies the absence of NOS-1 is related to greater muscle damage. However, further studies are needed, such as functional evaluation, to confirm this event

Automação do processo de avaliação de desempenho de tratores e implementos em campo

 Power machinery and implements are being used in many regions of Brazil to support different grain production systems without proper adaptation for working in different soil and weather conditions. To help farmers and manufacters solve problems related to equipments, a performance test needs to be working under a similar field condition. The objective of this research was to evaluate an in-field performance test, using an automatic data acquisition system. The engine power demanded was calculated indirectly, by using the engine rpm and the fuel consumption data on the tractor's calibration curves. The easiness of programming and operating the electronic instrumentation systems allowed to accomplish a great number of performance tests. The results showed very little difference between drawbar force measured and calculated obtaining 0.996 of correlation. The metodology used for in-field performance test showed to be a very powerfull tool to help farmers and the industry better manage their equipments and to improve performance.Em muitas regiões do Brasil, tratores e implementos agrícolas têm sido usados para dar suporte aos diferentes sistemas de produção, sem uma adequação própria às condições de solo e clima. Para auxiliar os fabricantes e agricultores na solução de problemas em máquinas e implementos agrícolas, há necessidade de verificar o desempenho desses equipamentos e estabelecer condições de testes semelhantes àquelas em que estiveram sendo usados. Este trabalho teve como objetivo avaliar o desempenho de um conjunto trator-implemento agrícola, no campo, utilizando um sistema eletrônico de medições. A potência demandada pelos equipamentos foi calculada indiretamente, com os dados de rotação do motor e consumo de combustível, nas curvas de calibração do trator. O uso de instrumentação eletrônica permitiu que se tivesse uma grande capacidade de trabalho, em razão da facilidade de programação e operação da cadeia de medições. Os resultados de campo obtidos para esforço de tração líquida indicaram tendência de comportamento similar à dos calculados por meio de fórmulas, com uma correlação de 0,996. A metodologia utilizada nos testes de desempenho operacional do conjunto trator-implemento no campo indicou ser uma ferramenta de bastante utilidade, tanto para os usuários dos equipamentos, quanto para os fabricantes

Acute Effect of Whole-Body Vibration in a Female Dog with Metritis

Background: Whole-body vibration is a modality of exercise used in humans for therapeutic purposes or to increase physical performance. In veterinary medicine there are only a few reports on the use of this technology. The Whole-body vibration derivate from vibrating rhythmic movements caused by vibrating platforms. Vibrating platforms are used over 30 years in the treatment and prevention of injuries and other debilitating conditions in humans. This paper aims to describe the first report of a possible spontaneous opening of the cervix in a female dog with purulent metritis by Whole-body vibration using a platform vibration.Case: A sexually intact female American Pit Bull Terrier dog showed an acute effect after a single session of Whole- body vibration training. Physical examination and physiological parameters were within normal. Slight edema of the vulva was observed without signs of discharge. Complete blood cell count, serum chemistry and urinalysis yielded no significant abnormal findings. An enlarged uterus with content was observed during a transabdominal ultrasound. Despite this, a vibrating-platform session was performed during 15 min. A purulent vulvar discharge was observed 6 min. after Whole-body vibration exercise and remained continuous through session. After the Whole-body vibration exercise, the dog was treated with cephalexin for 15 days. Escherichia coli were isolated from vaginal discharge culture. Seven days after the Whole-body vibration session, no signs of vulvar edema or discharge were observed. A complete blood count, serum chemistry, urinalysis and uterus ultrasound showed no abnormalities. Ovariohysterectomy by minimally invasive technique was performed and was observed cysts in both ovaries. The patient was placed under general anesthesia withisoflurane/O2. The premedication used was carprofen, morphine sulphate, acepromazine and diazepam. The induction was with propofol. An open ventral midline celiotomy an ovariohysterectomy was performed. Ten days after the surgery the dog showed no abnormality. After recovered from anesthesia, the patient was discharged with meloxican and tramadol for postoperative inflammation and pain management. According to the owner, the immediate postoperative period was without any problems. The histopathology examination confirmed the presence of right ovary cystic and left uterine horn cystic, and uterine hyperplasia. Four months after the surgery, the dog returned for evaluation and the owner reported that patient had been doing well, with normal appetite, normal urination and defecation.Discussion: The Whole-body vibration is not yet fully understood and it is important that the patient is clinically evaluated before the Whole-body vibration session. Whole-body vibration is a modality of exercise used in humans for therapeutic purposes or to increase physical performance. Otherwise, in veterinary medicine there are only a few reports on the use of this technology. In the present report there were no clinical signs that indicated uterine infection. In human patients there are reports that showed any clinical signs of diseases before use the Whole-body vibration. The ultrasound exam had been done because was part of another study and showed an enlarged uterus. Therefore, the cervix opening after a single Whole- body vibration training may be considered a positive side effect of Whole-body vibration in female dogs since this event helped to diagnose the disease. Although, the effects of WBV on reproductive organs and endocrine system are not clear

Evolution of the patella and patelloid in marsupial mammals

The musculoskeletal system of marsupial mammals has numerous unusual features beyond the pouch and epipubic bones. One example is the widespread absence or reduction (to a fibrous “patelloid”) of the patella (“kneecap”) sesamoid bone, but prior studies with coarse sampling indicated complex patterns of evolution of this absence or reduction. Here, we conducted an in-depth investigation into the form of the patella of extant marsupial species and used the assembled dataset to reconstruct the likely pattern of evolution of the marsupial patella. Critical assessment of the available literature was followed by examination and imaging of museum specimens, as well as CT scanning and histological examination of dissected wet specimens. Our results, from sampling about 19% of extant marsupial species-level diversity, include new images and descriptions of the fibrocartilaginous patelloid in Thylacinus cynocephalus (the thylacine or “marsupial wolf”) and other marsupials as well as the ossified patella in Notoryctes ‘marsupial moles’, Caenolestes shrew opossums, bandicoots and bilbies. We found novel evidence of an ossified patella in one specimen of Macropus rufogriseus (Bennett’s wallaby), with hints of similar variation in other species. It remains uncertain whether such ossifications are ontogenetic variation, unusual individual variation, pathological or otherwise, but future studies must continue to be conscious of variation in metatherian patellar sesamoid morphology. Our evolutionary reconstructions using our assembled data vary, too, depending on the reconstruction algorithm used. A maximum likelihood algorithm favours ancestral fibrocartilaginous “patelloid” for crown clade Marsupialia and independent origins of ossified patellae in extinct sparassodonts, peramelids, notoryctids and caenolestids. A maximum parsimony algorithm favours ancestral ossified patella for the clade [Marsupialia + sparassodonts] and subsequent reductions into fibrocartilage in didelphids, dasyuromorphs and diprotodonts; but this result changed to agree more with the maximum likelihood results if the character state reconstructions were ordered. Thus, there is substantial homoplasy in marsupial patellae regardless of the evolutionary algorithm adopted. We contend that the most plausible inference, however, is that metatherians independently ossified their patellae at least three times in their evolution. Furthermore, the variability of the patellar state we observed, even within single species (e.g. M. rufogriseus), is fascinating and warrants further investigation, especially as it hints at developmental plasticity that might have been harnessed in marsupial evolution to drive the complex patterns inferred here

Mensuração das dimensões cardíacas pelo método VHS (Vertebral Heart Size) em cães

It is imortant to be able to evaluate the cardiact size . In 1995, was proposed by Buchanan and Bücheler a new way to measure the dimensions of the cardiac silhouette. This study aims to gather several studies of the cardiac dimensions by VHS method in the right lateral recumbency of of different breeds of dogs in a single work, facilitating the assess to information. Deep chest breeds have a smaller mean VHS value than the barrel chest breeds, showing that there is a variation of the VHS values acoording to the thoracic conformation of each breed, which indicates that there is a need to recognize the specific VHS values according to the breed standard to not to interpret the cardiac measurement in a wrong way through not compatible reference values for the animal. There is the individual variation that should be taken into accont at the time of measurement. The VHS is a helper method to a radiographic evaluation of the heart, and the ultrasonography remains the gold standard for the evaluation of the cardiac chambers and other changes of the heartÉ importante ser capaz de avaliar o tamanho cardíaco radiograficamente. Em 1995, foi proposta por Buchanan e Bücheler uma nova forma de se avaliar as dimensões da silhueta cardíaca. O presente estudo tem por objetivo reunir diversos estudos das dimensões cardíacas pelo método VHS em decúbito lateral direito de diferentes raças de cães em um único trabalho, facilitando o acesso às informações. Raças de tórax profundo apresentaram um valor médio de VHS menor que as raças de tórax em formato de barril, mostrando que há variação dos valores de VHS de acordo com a conformação torácica de cada raça, o que indica que existe a necessidade de se conhecer os valores específicos de VHS de acordo com o padrão racial para que não se interprete a mensuração cardíaca de forma errônea através de valores de referência não compatíveis com o animal. Existe a variação individual que também deve ser levada em consideração no momento da mensuração. O VHS é um método auxiliar para a avaliação radiográfica do coração, e a ultrassonografia continua sendo o padrão ouro para avaliação das câmaras cardíacas e outras alterações do coraçã

Automação do processo de avaliação de desempenho de tratores e implementos em campo Electronic instrumentation use on field performance test of agricultural tractors and implements

Em muitas regiões do Brasil, tratores e implementos agrícolas têm sido usados para dar suporte aos diferentes sistemas de produção, sem uma adequação própria às condições de solo e clima. Para auxiliar os fabricantes e agricultores na solução de problemas em máquinas e implementos agrícolas, há necessidade de verificar o desempenho desses equipamentos e estabelecer condições de testes semelhantes àquelas em que estiveram sendo usados. Este trabalho teve como objetivo avaliar o desempenho de um conjunto tratorimplemento agrícola, no campo, utilizando um sistema eletrônico de medições. A potência demandada pelos equipamentos foi calculada indiretamente, com os dados de rotação do motor e consumo de combustível, nas curvas de calibração do trator. O uso de instrumentação eletrônica permitiu que se tivesse uma grande capacidade de trabalho, em razão da facilidade de programação e operação da cadeia de medições. Os resultados de campo obtidos para esforço de tração líquida indicaram tendência de comportamento similar à dos calculados por meio de fórmulas, com uma correlação de 0,996. A metodologia utilizada nos testes de desempenho operacional do conjunto trator-implemento no campo indicou ser uma ferramenta de bastante utilidade, tanto para os usuários dos equipamentos, quanto para os fabricantes.Power machinery and implements are being used in many regions of Brazil to support different grain production systems without proper adaptation for working in different soil and weather conditions. To help farmers and manufacters solve problems related to equipments, a performance test needs to be working under a similar field condition. The objective of this research was to evaluate an in-field performance test, using an automatic data acquisition system. The engine power demanded was calculated indirectly, by using the engine rpm and the fuel consumption data on the tractor's calibration curves. The easiness of programming and operating the electronic instrumentation systems allowed to accomplish a great number of performance tests. The results showed very little difference between drawbar force measured and calculated obtaining 0.996 of correlation. The metodology used for in-field performance test showed to be a very powerfull tool to help farmers and the industry better manage their equipments and to improve performance