Effects of physical training on the immune system in diabetic rats

Aims: This study aims to investigate the influence of physical training on the immune system of diabetic rats. Materials and Methods: Adult male Wistar rats were distributed into Sedentary Control (SC), Trained Control (TC), Sedentary Diabetic (SD) and Trained Diabetic (TD) groups were used. Diabetes was induced by alloxan (32 mg/bw-i.v.). Training protocol consisted of swimming, at 32 18C, one hour/day, five days/week, supporting an overload equivalent to 5 of the body weight, during four weeks. At the end of the experiment the rats were sacrificed by decapitation and blood samples were collected for glucose, insulin, albumin, hematocrit determinations, total and differential leukocyte counting. Additionally, liver samples for glycogen analyses were obtained. Results: The results were analyzed by one way at a significance level of 5. Diabetes reduced blood insulin, liver glycogen stores and increased blood glucose and neutrophil count. Physical training restored glycemia, liver glycogen levels, neutrophils and lymphocytes count in diabetic rats. Conclusions: In summary, physical training was able to improve metabolic and immunological aspects in the experimental diabetic rats.

Epigenetic Silencing of CRABP2 and MX1 in Head and Neck Tumors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morphological, morphometrical and ultrastructural characterization of Phrynops geoffroanus' (Testudines: Chelidae) blood cells, in different environments

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Distinct localization of T cell Agrin during antigen presentation - evidence for the expression of Agrin receptor(s) in antigen-presenting cells

Agrin is over-expressed by activated and autoimmune T cells, and synergizes with the T cell receptor (TCR) to augment cell activation. In the present study, we show that Agrin accumulates to distinct areas of the plasma membrane and that cell activation causes its redistribution. During antigen presentation, Agrin primarily accumulates to the periphery of the mature immunological synapse, mostly in lamellipodia-like protrusions that wrap around the antigen-presenting cell and, conversely, anti-Agrin sera induced a significant redistribution of TCR at the plasma membrane. We also provide evidence for the expression of Agrin receptors in peripheral blood monocytes, dendritic cells and a fraction of B cells. Interestingly, interferon-a treatment, which induces the expression of Agrin in T cells, also augmented Agrin binding to monocytes. Stimulation of monocytes with recombinant Agrin induced the clustering of surface receptors, including major histocompatibility complex class II, activation of intracellular signalling cascades, as well as enhanced dsRNA-induced expression of pro-inflammatory cytokines interleukin-6 and tumour necrosis factor-a. Collectively, these results confirm the location of Agrin at the immunological synapse between T cells and antigen-presenting cells and justify further characterization of its receptors in the immune system.

Annexin 1 mimetic peptide protects against renal ischemia/reperfusion injury in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Alterações cromossômicas causadas pela radiação dos monitores de vídeo de computadores

OBJETIVO: em decorrência dos questionamentos sobre o efeito deletério das radiações emitidas pelo campo eletromagnético (CEM) dos tipos ELF (extremely low frequency) e VLF (very low frequency) transmitidas pelos monitores de vídeo dos computadores (CRT), foi avaliada a freqüência de anomalias cromossômicas estruturais e a cinética do ciclo celular em indivíduos expostos por seu trabalho à radiação dos CRT. MÉTODOS: A pesquisa de aberrações cromossômicas foi realizada em 2.000 metáfases de primeira divisão celular obtidas de culturas de 48h de linfócitos de sangue venoso periférico de dez indivíduos expostos ao CRT (grupo E) e de dez controles (grupo C). A cinética do ciclo celular foi pesquisada pelos índices mitótico (IM) e de proliferação celular (IPC). RESULTADOS: A análise estatística evidenciou freqüências significativamente maiores de metáfases com anomalias cromossômicas (E=5,9%; C=3,7%) e anomalias/célula (E=0,066±0,026; C=0,040±0,026) nos indivíduos expostos aos CRTs. As alterações citogenéticas mais comuns foram as quebras cromatídicas, com freqüência de 0,034±0,016 no grupo E e de 0,016±0,015 no grupo C. As freqüências de IM e IPC não apresentaram diferenças significantes entre os grupos avaliados. CONCLUSÕES: Os resultados sugerem um efeito genotóxico do CEM emitido pelos CRTs devido à freqüência mais elevada de quebras cromatídicas, enfatizando a necessidade de haver um número maior de estudos com diferentes técnicas que vise a investigar a ação do CEM sobre o material genético.

Influence of the Bilayer Composition on the Binding and Membrane Disrupting Effect of Polybia-MP1, an Antimicrobial Mastoparan Peptide with Leukemic T-Lymphocyte Cell Selectivity

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

In vitro effects of calcium hydroxide and polymyxin B on endotoxins in root canals

Objectives. To evaluate the effects of intracanal medicaments on endotoxins in root canals.Methods. Seventy-five freshly extracted maxillary incisors were used in this study. The crowns of teeth were sectioned near the CEJ in order to standardize the root length to 14 mm. The root canals were instrumented to an apical size #50 file and irrigated with 1% sodium hypochlorite solution and sterilized with 60 Co gamma irradiation. Standardized suspension containing Escherichia coli endotoxin was inoculated into the 60 root canals. The specimens were randomly assigned to 5 groups (n=15), according to the intracanal medicament used: (G1) calcium hydroxide; (G2) polymyxin B; (0) combination neomycin-potymyxin B-hydrocortisone; (G4) positive control (no intracanal medicament); (G5) negative control (no endotoxin and no intracanal medicament). After 7 days, the detoxification of endotoxin was evaluated by Limulus lysate assay and antibody production in B-tymphocytes culture.Results. Groups 1, 2 and 5 presented the best results by Limulus lysate and were significantly different to groups 3 and 4 (p<0.05). Stimulation of antibodies production in cell culture by groups 1 and 6 was smaller and statistically different than groups 2, 3, 4 and 5 (p<0.05). Groups 2 and 5 induced a small increase in the antibodies production in relation to the groups 1 and 6. Groups 3 and 4 induced a significant increase of antibodies production (p<0.05).Conclusions. The calcium hydroxide and polymyxin B intracanal medicaments detoxified endotoxin in root canals and altered the properties of LPS to stimulate the antibody production by B-Lymphocytes. The combination neomycin-polymyxin B-hydrocortisone did not detoxified endotoxin. (C) 2004 Elsevier Ltd. All rights reserved.

Detecção de anticorpo anticoração em camundongos Balb/c imunizados com Streptococcus mutans

Anticorpos para antígenos cardíacos foram analisados por ELISA em 14 soros de camundongos Balb/c hiperimunizados com Streptococcus mutans, inativado pelo formaldeído. Os níveis de anticorpos da classe IgG anticoração e antimiosina elevaram-se significativamente nos animais imunizados quando comparados com os controles, especialmente no grupo A, imunizado e reestimulado com antígenos solúveis de S. mutans. Neste grupo, os resultados do Western Blot mostraram reatividade com miosina cardíaca e uma banda de 35 kDa. A análise histológica dos corações dos animais do grupo B, imunizado e reestimulado com antígenos de superfície do microrganismo, demonstrou a presença de degeneração celular, tipo hidrópica e hialina e focos inflamatórios constituídos de linfócitos e macrófagos no miocárdio e pericárdio. Os resultados deste trabalho reforçam a hipótese da existência de mimetismo antigênico entre tecido cardíaco e S. mutans e chamam a atenção para o risco de desenvolvimento de anticorpos reativos com antígenos próprios induzidos por vacina anticárie com componentes estreptocócicos.

Epstein-Barr virus-associated gastric carcinoma in Brazil: comparison between in situ hybridization and polymerase chain reaction detection

Epstein-Barr virus (EBV) has been associated with 10% of gastric carcinomas. The aim of this study was to determine the frequency of EBV in gastric carcinomas in Brazil assessed by in situ hybridization (ISH) and PCR, which would contribute to the characterization of the clinical and pathological aspects of EBV-associated gastric carcinomas. One hundred and ninety-two gastric carcinoma cases were collected at hospitals in two Brazilian states. Seventy-three out of 151 cases were PCR(+), while 11/160 cases were ISH(+). Nine out of eleven ISH(+) cases displayed a diffuse staining pattern and 2 out of 11 a focal pattern. Both techniques showed that the EBV(+) cases were characterized by their association with males, older patients, lower gastric region, intestinal type, advanced stage and poorly to moderately differentiated tumors. The concordance between the two techniques was 55.8% (Cohen's kappa index = 0.034). Four cases were ISH(+)/PCR(-), while 49 cases were PCR(+)/ISH(-). Only two cases showed stained lymphocytes by ISH and one of them was PCR(-). The observed discrepancy between the two techniques could not be explained just by the elevated accuracy of PCR. ISH(+)/PCR(-) carcinomas may be encountered if EBV is not present in the whole tumor tissue or if there are polymorphisms in the sequences of the viral genome amplified. on the other hand, the high frequency of PCR(+) results associated with the absence of ISH staining in lymphocytes and/or tumors cells suggests that the virus may be present in tumor cells or other cell types without expressing EBER1, the target of the ISH technique.

Efeitos do treinamento físico de alta intensidade sobre os leucócitos de ratos diabéticos

Estudos têm demonstrado que o exercício físico regular melhora as condições do diabetes, facilitando a captação periférica da glicose e o metabolismo de glicogênio, proteínas, etc. Por outro lado, pouco se conhece sobre os efeitos do exercício intenso em diabéticos, principalmente com relação ao sistema imune desses organismos. O presente estudo teve como objetivo verificar os efeitos de um treinamento físico de alta intensidade sobre a contagem total e diferencial de leucócitos em ratos diabéticos. Ratos machos jovens Wistar foram distribuídos em quatro grupos: controle sedentário (CS), controle treinado (CT), diabético sedentário (DS) e diabético treinado (DT). O diabetes foi induzido por aloxana (35mg/kg de peso corporal). Durante seis semanas os animais dos grupos CT e DT realizaram um protocolo de treinamento físico, que consistiu na realização de quatro séries de 10 saltos (intercaladas por um minuto de intervalo) em piscina, com o nível da água correspondendo a 150% do comprimento corporal e sobrecarga equivalente a 50% da massa corporal dos animais. Ao final do período experimental, amostras de sangue foram coletadas para a contagem total e diferencial dos leucócitos. Os resultados foram avaliados estatisticamente por ANOVA com um nível de significância de 5%. A glicemia foi aumentada entre os diabéticos e a insulinemia diminuída. Não foram observadas diferenças significativas na contagem diferencial dos linfócitos, neutrófilos, eosinófilos e contagem total de leucócitos entre os grupos estudados. Houve aumento dos monócitos entre os treinados (CS = 10,0 ± 4,5, CT* = 25,4 ± 7,9, DS = 19,75 ± 7,4, DT* = 25,8 ± 4,4%). O peso relativo do timo foi reduzido pelo treinamento e pelo diabetes (CS = 125,0 ± 37,7, CT* = 74,6 ± 8,2, DS* = 47,5 ± 12,2, DT* = 40,1 ± 16,9mg/100g). Esses resultados permitem concluir que o treinamento físico de alta intensidade não alterou o estado geral do diabetes, mas aumentou os monócitos, o que pode representar um efeito positivo sobre a resposta imunológica desses animais.

Detecção do vírus Epstein-Barr (EBV) em adenocarcinomas gástricos procedentes dos estados do Ceará e de São Paulo

INTRODUÇÃO: O vírus Epstein-Barr (EBV) está associado a cerca de 10% dos adenocarcinomas gástricos, representando mais de 50 mil casos por ano no mundo. Apesar dos estudos realizados em várias partes do mundo, alguns aspectos clinicopatológicos permanecem controversos. OBJETIVOS: O presente estudo teve como objetivo analisar as características clinicopatológicas de casos de adenocarcinomas gástricos procedentes dos estados de São Paulo e Ceará, correlacionando-os com a detecção de EBV. MATERIAIS E MÉTODOS: Foram obtidos 192 casos de adenocarcinomas gástricos de hospitais dos estados de São Paulo e do Ceará, dos quais 160 foram submetidos à técnica de RNA-hibridização in situ para detecção de EBV. RESULTADOS: Dos 160 casos, 11 (6,9%) foram EBV-positivo, exibindo intensa marcação nuclear em células tumorais. Destes, dois casos também apresentaram linfócitos infiltrados marcados. Não encontramos marcação em tecido normal ou pré-neoplásico. São Paulo e Ceará apresentaram as frequências 3/60 (5%) e 8/100 (8%), respectivamente, e maior relação do EBV com indivíduos do sexo masculino, de idade avançada, com tumores do tipo intestinal, de estadiamento elevado e grau pouco a moderadamente diferenciado. Os casos do Ceará exibiram aumento relativo de tumores EBV(+) localizados na cárdia, enquanto os casos de São Paulo demonstraram aumento naqueles localizados no corpo gástrico. CONCLUSÃO: A frequência de tumores EBV(+) do presente estudo situa-se nos valores descritos na literatura mundial. Entre os achados, um deles não encontra paralelo na literatura mundial e refere-se ao elevado percentual de tumores EBV(+) no corpo gástrico observado nos casos de São Paulo.

Clinical and hematological alterations in dogs during experimental envenomation with Crotalus durissus terrificus venom and treated with antiophidic serum

The present work aimed to evaluate the clinical and hematological aspects during experimental envenomation by Crotalus durissus terrificus in dogs treated with different antiophidic serum doses. Sixteen dogs were divided into two groups of eight animals each. Group I received 1mg/kg venom subcutaneously and 30mg antiophidic serum intravenously; Group II received 1mg/kg venom subcutaneously and 60mg antiophidic serum intravenously. In the clinical evaluation, we observed ataxia, moderate sedation, dilated pupils, sialorrhea, flaccid paralysis of mandibular muscles, and discreet edema at the site of venom inoculation. Evaluating red and white blood cells, we observed a decrease of hemoglobins, globular volume and erythrocytes, and an increase of plasmatic proteins, leukocytes, neutrophils, monocytes and lymphocytes. Clotting time increased and there was blood incoagulability with return to normal clotting time six hours after antiophidic serum administration. Animals treated with six antiophidic serum flasks had a faster recovery than the animals that received three serum flasks.

Response activity of alveolar macrophages in pulmonary dysfunction caused by Leptospira infection

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resistance to experimental autoimmune encephalomyelitis development in Lewis rats from a conventional animal facility

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory disease of the brain and spinal cord that is mediated by CD4+ T lymphocytes specific to myelin components. In this study we compared development of EAE in Lewis rats from two colonies, one kept in pathogen-free conditions (CEMIB colony) and the other (Botucatu colony) kept in a conventional animal facility. Female Lewis rats were immunized with 100 µl of an emulsion containing 50 µg of myelin, associated with incomplete Freund's adjuvant plus Mycobacterium butyricum. Animals were daily evaluated for clinical score and weight. CEMIB colony presented high EAE incidence with clinical scores that varied from three to four along with significant weight losses. A variable disease incidence was observed in the Botucatu colony with clinical scores not higher than one and no weight loss. Immunological and histopathological characteristics were also compared after 20 days of immunization. Significant amounts of IFN-gamma, TNF-alpha and IL-10 were induced by myelin in cultures from CEMIB animals but not from the Botucatu colony. Significantly higher levels of anti-myelin IgG1 were detected in the CEMIB colony. Clear histopathological differences were also found. Cervical spinal cord sections from CEMIB animals showed typical perivascular inflammatory foci whereas samples from the Botucatu colony showed a scanty inflammatory infiltration. Helminths were found in animals from Botucatu colony but not, as expected, in the CEMIB pathogen-free animals. As the animals maintained in a conventional animal facility developed a very discrete clinical, and histopathological EAE in comparison to the rats kept in pathogen-free conditions, we believe that environmental factors such as intestinal parasites could underlie this resistance to EAE development, supporting the applicability of the hygiene hypothesis to EAE.