Beneficial effect of Lactococcus lactis NCC 2287 in a murine model of eosinophilic esophagitis.

BACKGROUND: Eosinophilic esophagitis (EoE) is a severe inflammatory disease of the esophagus which is characterized histologically by an eosinophilic infiltration into the esophageal tissue. The efficacy of probiotics in the context of atopic diseases has been well investigated but, to date, there has been no study which has evaluated probiotic effects on EoE inflammation. This study sought to identify a probiotic which improves esophageal inflammation in experimental EoE. METHODS: Two candidate probiotics, Lactococcus lactis NCC 2287 and Bifidobacterium lactis NCC 2818, were tested in a murine model of EoE elicited by epicutaneous sensitization with Aspergillus fumigatus protein extract. Administration of bacterial strains in drinking water was used, respectively, as a preventive or treatment measure, or continuously throughout the study. Inflammatory parameters were assessed in the esophagus, skin, and lungs after allergen challenge. RESULTS: In this EoE model, supplementation with L. lactis NCC 2287 significantly decreased esophageal and bronchoalveolar eosinophilia but only when given as a therapeutic treatment. No significant effect on eosinophilia was observed when NCC 2287 was given as a preventive or a continuous intervention. NCC 2287 supplementation had no significant effect on immunoglobulin levels, skin symptom scores, or on transepidermal water loss. Supplementation with another probiotic, B. lactis NCC 2818, had no significant effect on esophageal eosinophilia. CONCLUSION: We identified a L. lactis strain, able to attenuate esophageal eosinophilic inflammation in a preclinical model of EoE. This effect is strain specific and depends on the timing and duration of bacterial supplementation. Confirmation of these observations in human clinical trials is warranted.

Genetic immunization and comprehensive screening approaches in HLA-A2 transgenic mice lead to the identification of three novel epitopes in hepatitis C virus NS3 antigen.

Interferon-gamma (IFN gamma)-producing CD8+ T cells have been shown to play a key role in the control or eradication of hepatitis C virus (HCV) infections. In particular, T cells specific of the non-structural protein 3 (NS3) are often associated with control of viremia. The aim of the study was to identify novel HLA-A2 restricted CD8+ T cell epitopes specific of NS3 using a combination of comprehensive approaches. HLA-A2.1 transgenic mice were immunized with a DNA vaccine optimized for NS3 specific epitope presentation and induced CD8+ T cell reactivity was screened using 42 algorithm-predicted peptides as well as a library of 78 overlapping 15-mer peptides spanning the whole protein. Three epitopes mapping within the NS3 protease (GLL: aa 1038-1047) or helicase (ATL: aa 1260-1268 and TLH: aa 1617-1625) were identified. These epitopes, which display similar and high in vitro binding capacities to soluble HLA-A2 molecules, are able to induce either cytotoxic T lymphocytes (CTL) and/or IFN gamma-producing T cells. Comparative in vitro target cell sensitization studies revealed a higher immunogenicity of the GLL peptide as compared with both ATL and TLH peptides. This peptide was capable to recall in vitro HCV-specific IFN gamma and IL-10-producing T cells from peripheral blood mononuclear cells (PBMC) of chronically infected patients. These data increase the pool of NS3-specific CD8+ T cell epitopes available to analyze HCV associated immunity and could contribute to the design and evaluation of candidate vaccines.

Localization of fission yeast type II myosin, Myo2, to the cytokinetic actin ring is regulated by phosphorylation of a C-terminal coiled-coil domain and requires a functional septation initiation network.

Myo2 truncations fused to green fluorescent protein (GFP) defined a C-terminal domain essential for the localization of Myo2 to the cytokinetic actin ring (CAR). The localization domain contained two predicted phosphorylation sites. Mutation of serine 1518 to alanine (S(1518)A) abolished Myo2 localization, whereas Myo2 with a glutamic acid at this position (S(1518)E) localized to the CAR. GFP-Myo2 formed rings in the septation initiation kinase (SIN) mutant cdc7-24 at 25 degrees C but not at 36 degrees C. GFP-Myo2S(1518)E rings persisted at 36 degrees C in cdc7-24 but not in another SIN kinase mutant, sid2-250. To further examine the relationship between Myo2 and the SIN pathway, the chromosomal copy of myo2(+) was fused to GFP (strain myo2-gc). Myo2 ring formation was abolished in the double mutants myo2-gc cdc7.24 and myo2-gc sid2-250 at the restrictive temperature. In contrast, activation of the SIN pathway in the double mutant myo2-gc cdc16-116 resulted in the formation of Myo2 rings which subsequently collapsed at 36 degrees C. We conclude that the SIN pathway that controls septation in fission yeast also regulates Myo2 ring formation and contraction. Cdc7 and Sid2 are involved in ring formation, in the case of Cdc7 by phosphorylation of a single serine residue in the Myo2 tail. Other kinases and/or phosphatases may control ring contraction.

Detection of hepatitis B virus DNA by the polymerase chain reaction in anti-HBE positive chronic hepatitis B patients

A deteccao do DNA do Virus da Hepatite B pela Reacao em cadeia Polimerase (PCR) foi comparada com os outros marcadores sorologicos virais (AgHBs, AgHBe e anti-HBe) numa serie de 49 pacientes com hepatite cronica B, incluindo 12 que apresentaram clareamento espontaneo do AgHBs. Nenhum caso AgHBs negativo foi PCR positivo, mas 33/37 (89,2 por cento) dos casos AgHBs positivos foram PCR positivos (p<0,0001). Entre as amostras AgHBs positivas, 9 foram AgHBe positivas e anti-HBe negativas, todas elas PCR positivas....

A anestesia no Sarah - Instituto Nacional de Medicina do Aparelho Locomotor: exames laboratoriais pré-operatórios / Anesthesia in the SARAH - Instituo Nacional de Medicina do Aparelho Locomotor: preoperatve screening test

De 1.500 pacinetes operados em 1981, 56 por cento tinham o estado físico ASA I e 40 por cento ASA II. Näo nos parecia correto que estes pacientes tivessem os mesmos exames pré-operatórios que os 4 por cento restantes (ASA III e IV). Para a escolha de determinado exame uma rotina pré-operatória devem ser consideradas: a prevalência das doenças, a sensibilidade, a especificidade, o custo, o risco e o benefício do exame. Baseados na estatística de Roizen nossos pacientes foram divididos em 2 grandes grupos, por uma comissäo médica multidisciplinar, no seguinte esquema de exames pré-operatórios: 0 a 45 anos Sumário de urina, hemograma completo, coagulograma, classificaçäo sangüínea. Acima de 45 anos: Sumário de urina, hemograma completo, coagulograma, classificaçäo sangüínea, raios X de tórax, Eletrocardiograma, uréia, creatinina, glicose séricas, Reaçäo de Machado Guerreiro. Recomendou-se o bom senso no pedido de outros exames principalmente na faixa etária abaixo de 45 anos se a anamnese e o exame físico assim os indicassem. Trataram-se todos os pacientes com vermífugo polivalente (mebendazol) pela freqüência d exames falso-negativos, com pedido de exame parasitológico após o tratamento (para excluir-se giardíase, estrongiloidíase, esquistossomiase e amebíase). Em conclusäo, esta orientaçäo, em funcionamento há 7 meses näo aumentou a morbidade transoperatória e pós-operatória imediata em 820 pacientes e contribuiu para reduzir o custo hospitalar e o sacrifício de nossos pacientes