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Activation of astrocytes or astrogliosis is a prominent component of the inflammatory response and an indicator of injury in the brain. These astrocytes produce a large array of inflammatory mediators, growth and neuroprotective factors. This study was an investigation from astrocyte (GFAP) response and TGF- ? 1 involvement during VSV acute encephalitis using immunohistochemistry to verify relation between astrocytes and TGF- ? 1. Animals developed symptoms around 6 th day after VSV inoculation. Viral proteins were mainly detected at olfactory bulb, ventricular cell layer and disseminated to hippocampus, mesencephalon and diecephalon areas and brain stem. Also at 6 th day post inoculation GFAP and TGF- ? 1 staining was observed in good association with virus-detected areas of brain. However, in mice with severe symptoms we observed reduction in the intensity of GFAP labeling at the same areas where TGF- ? 1 upregulation was observed. These areas show correlation with areas of necrosis and where are astrocytes with degenerative aspect. We observed TGF- ? 1 staining in damaged astrocytes, suggesting an effort of those cells in controlling inflammation in acute phase of VSV encephalitis.

O presente estudo teve por objetivo avaliar a capacidade de concentração plaquetária e sua correlação com os níveis do fator de crescimento TGF-B1, a presença de leucócitos e de hemácias nos diferentes protocolos utilizados na obtenção do plasma rico em plaquetas (PRP) de equinos, através do método manual. Dez equinos, sadios, com idade média de 7 anos (±2,39), pesando em média 500kg (±67,1) foram utilizados neste estudo. Os protocolos testados variaram na velocidade e no tempo nas duas centrifugações. As variáveis analisadas nas amostras de PRP foram: concentração de plaquetas, presença de leucócitos e hemácias, e níveis de TGF-β1 quantificados pelo teste ELISA. Os protocolos testados não diferiram na capacidade de concentração de plaquetas e nos níveis de TGF-β1. Entretanto, houve diferença significava entre o protocolo I e os demais por este apresentar maior número de hemácias e leucócitos nas amostras de PRP, sendo por esse motivo considerado um protocolo inadequado para processamento do volume de sangue utilizado. Os demais protocolos podem ser utilizados para obtenção de PRP terapêutico em equinos. PRP is plasma that contains a high numbers of platelets and growth factors in a small volume. The aim of this study was to evaluate seven different protocols to obtain PRP by the manual method according to their capacity to concentrate platelets, leukocyte and erythrocyte contamination and correlation between platelet count and TGF-β1 growth factor levels in PRP samples. Ten healthy horses with a mean age of 7 years (±2.39), weighing on average 500 kg (±67.1) were used in this study. The protocols tested varied according to the speed and time used at the two centrifugations. PRP samples were analyzed regarding platelet concentration, leukocyte and erythrocyte contamination and TGF-β1 levels quantified by ELISA. No significant differences among protocols were observed regarding the ability to concentrate platelets and TGF-β1. However, protocol I showed significantly higher erythrocyte and leukocyte counts in PRP samples than the other protocols, reason why it was considered inadequate for the volume of blood processed in this experiment. The remaining protocols are suitable for extracting PRP.

Introdução: Biomarcadores séricos têm sido usados no diagnóstico e avaliação da intensidade da fibrose na esquistossomose hepatoesplênica com resultados variados. Poucos estudos usaram a biópsia hepática na confirmação da presença de fibrose de Symmers e mais frequentemente utilizaram-se métodos de imagem como padrão ouro. Objetivo: Avaliamos neste estudo o papel do ácido hialurônico (AH), do YKL40 e do fator de crescimento e transformação 1 (TGF-1) no diagnóstico da esquistossomose mansônica hepatoesplênica e na avaliação da intensidade da fibrose hepática utilizando métodos de imagem e biópsia hepática cirúrgica. Metodologia: Sessenta pacientes com esquistossomose mansônica foram selecionados para o estudo. Trinta (Grupo 1) tinham esquistossomose hepatoesplênica e frequentavam o ambulatório de Doenças Infecciosas e Parasitárias da UFMG (estes pacientes não tinham evidência de infecção ativa pelo S. mansoni); 23 eram homens (76,7%), com média de idade de 39,2 anos (± 9,7). Outros 30 pacientes tinham esquistossomose hepatointestinal (Grupo 2) e eram provenientes de área endêmica de esquistossomose (ovos viáveis nas fezes); 15 eram homens (50%), com média de idade de 35,5 anos (±12,7). Todos foram submetidos a exame clínico, ultrassonografia abdominal e coleta de sangue para a realização dos marcadores séricos de fibrose. Os hepatoesplênicos (Grupo 1) submeteram-se a outros exames, como: marcadores sorológicos de hepatites B e C, endoscopia digestiva alta, ressonância magnética do abdome e biópsia hepática em cunha durante o ato cirúrgico para tratamento de hipertensão porta. Os marcadores séricos foram dosados pelo método de ELISA utilizando kits comerciais. Para a ultrassonografia usou-se o aparelho ALOKA SSD 1700 Dynaview e para a ressonância magnética o aparelho da General Eletric (GE) 1.5 tesla. Os fragmentos de fígado obtidos durante a cirurgia foram conservados em formol a 10% e a seguir emblocados em parafina. Os cortes de 5m (micrômetros) corados pela HE foram analisados à microscopia de luz. Para a histomorfometria usou-se o corante picrossírius vermelho e as áreas examinadas foram quantificadas usando-se o software Image-Pro-Plus. Os dados obtidos foram armazenados no programa EpiData 3.0 e analisados pelo software R Project for Statistical Computing, versão 2.14.0. Resultados: O ácido hialurônico (AH) e o YKL40 não tiveram valor no diagnóstico da fibrose hepática da esquistossomose (não separaram os hepatoesplênicos dos hepatointestinais), quando comparados aos métodos de imagem. A distribuição dos valores de TGF-1 foi maior no grupo de pacientes com esquistossomose hepatointestinal (p= 0,0001). A classificação da intensidade da fibrose hepática na histologia não coincidiu com os valores séricos dos marcadores avaliados neste estudo. Observou-se correlação moderada dos níveis séricos de AH com a histomorfometria (p= 0,006). Houve boa concordância entre os métodos de imagem e a biópsia hepática na classificação da intensidade da fibrose. Conclusões: O ácido hialurônico e o YKL40 não separaram os pacientes com fibrose dos pacientes sem fibrose hepática. O TGF-1 não teve importância como marcador de fibrose hepática nos hepatoesplênicos, mas apresentou níveis séricos significativamente mais elevados nos pacientes com esquistossomose hepatointestinal. Isso sugere que ele seja marcador de infecção ativa pelo S. mansoni. Os marcadores usados neste estudo não foram importantes na classificação da intensidade da fibrose hepática esquistossomótica. Introduction: Serum biomarkers have been used as a tool in the diagnosis and evaluation of liver fibrosis intensity in hepatosplenic schistosomiasis with variable results. Few studies have used liver biopsy in the confirmation of Symmers' fibrosis and more frequently physicians rely upon imaging techniques as gold standard in the evaluation of liver fibrosis severity. Objective: Herein we evaluated the importance of hyaluronic acid (HA), YKL-40 and transforming growth factor Beta1 (TGF-1) in the diagnosis of hepatosplenic schistosomiasis and in the evaluation of liver fibrosis intensity using imaging techniques and surgical wedge liver biopsy. Methodology: Sixty patients with schistosomiasis mansoni were selected for this study. Thirty (Group 1) had hepatosplenic schistosomiasis and they were attending the Centro de Treinamento e Referência em Doenças Infecciosas e Parasitárias, UFMG (patients, in this group, had no evidence of active Schistosoma mansoni infection); 23 were male (76.7%) in the mean age of 39.2 years (±9.7). Another 30 patients had hepatointestinal schistosomiasis (Group 2) and they came from endemic areas of Minas Gerais (with viable eggs in the stool); 15 were male (50.0%), mean age of 35.5 years (±12.7). All patients were submitted to clinical examination and abdominal ultrasound; a blood sample was collected and stored for further analysis. The hepatosplenic group was submitted to other tests, such as: serology for hepatitides B and C, upper digestive endoscopy, abdominal magnetic resonance and surgical liver wedge biopsy. The serum markers of fibrosis were measured using commercial kits. For ultrasound an ALOKA SSD 1700 Dynaview apparatus was used and for magnetic resonance a GE 1.5 tesla instrument. Liver fragments obtained during surgery were fixed in 10% buffered formalin and afterwards embedded in paraffin wax. Five m (micrometers) slices were stained using Hematoxylin-Eosin and examined under light microscopy. Other fragments were stained with Picrosirius red and portal tracts were selected by the examiner and quantified by the software Image-Pro-Plus. Collected data were stored in EpiData 3.1 and the software R Project for Statistical Computing, 2.14.0 version was used for statistical analysis. Results: Both hyaluronic acid and YKL40 had no value in the diagnosis of liver fibrosis (they did not separate patients with hepatosplenic from hepatointestinal schistosomiasis) when imaging techniques were used for liver fibrosis identification. Serum levels of TGF-B1 were higher in the sera of patients with hepatointestinal schistosomiasis compared to patients with the hepatosplenic form of the disease. Intensity of liver fibrosis classified by histology did not coincide with serum levels of the biomarkers evaluated in this study. There was moderate correlation between serum levels of hyaluronic acid when it was compared to histomorphometry (p=0.006). There was a good concordance between imaging techniques and liver biopsy in the classification of liver fibrosis intensity. Conclusions: Hyaluronic acid and YKL40 were not useful as a marker of liver fibrosis in our study. TGF- 1, also, was not a good marker of liver fibrosis but its serum levels were significantly higher in patients with hepatointestinal schistosomiasis as compared to hepatosplenics. Therefore, TGF-1 may be a marker of active S. mansoni infection. The biomarkers used in the present study were not important in classifying schistosomiasis liver fibrosis severity.