Induction of TNF-alfa and CXCL-2 mRNAs in different organs of mice infected with pathogenic Leptospira

The role of innate immune response in protection against leptospirosis is poorly understood. We examined the expression of the chemokine CXCL2/MIP-2 and the cytokine TNF-alpha. in experimental resistant and susceptible mice models, C3H/HeJ, C3H/HePas and BALB/c strains, using a virulent strain of Leptospira interrogans serovar Copenhageni. Animals were infected intraperitoneally with 107 cells and the development of the disease was followed. Mortality of C3H/HeJ mice was observed whereas C3H/HePas presented jaundice and BALB/c mice remained asymptomatic. The infection was confirmed by the presence of leptospiral DNA in the organs of the animals, demonstrated by PCR. Sections of the organs were analyzed, after H&E stain. The relative expression of mRNA of chemokine CXCL2/MIP-2 and cytokine TNF-alpha was measured in lung, kidney and liver of the mice by qPCR. The concentrations of these proteins were measured in extracts of tissues and in serum of the animals, by ELISA. Increasing levels of transcripts and protein CXCL2/MIP-2 were detected since the first day of infection. The highest expression was observed at third day of infection in kidney, liver and lung of BALB/c mice. In C3H/HeJ the expression of CXCL2/MIP-2 was delayed, showing highest protein concentration in lung and kidney at the 5th day. Increasing in TNF-alpha transcripts were detected after infection, in kidney and liver of animals from the three mice strains. The expression of TNF-alpha protein in C3H/HeJ was also delayed, being detected in kidney and lung. Our data demonstrated that Leptospira infection stimulates early expression of CXCL2/MIP-2 and TNF-alpha in the resistant strain of mice. Histological analysis suggests that the expression of those molecules may be related to the influx of distinct immune cells and plays a role in the naturally acquired protective immunity. (C) 2012 Elsevier Ltd. All rights reserved.

Re-treatment of previous non-responders and relapsers to interferon plus ribavirin with peginterferon alfa-2a (40KD), ribavirin +/- amantadine in patients with chronic hepatitis C: randomized multicentre clinical trial

Introduction. A large number of patients with chronic hepatitis C have not been cured with interferon-based therapy. Therefore, we evaluated the efficacy of amantadine combined with the standard of care (pegylated interferon plus ribavirin) in patients who had not responded to or had relapsed after 24 weeks of treatment with conventional interferon plus ribavirin. Material and methods. Patients stratified by previous response (i.e., non-response or relapse) were randomized to 48 weeks of open-label treatment with peginterferon alfa-2a (401(D) 180 pg/week plus ribavirin 1,000/1,200 mg/day plus amantadine 200 mg/day (triple therapy), or the standard of care (peginterferon alfa-2a [40KD] plus ribavirin). Results. The primary outcome was sustained virological response (SVR), defined as undetectable hepatitis C virus RNA in serum (< 50 IU/mL) at end of follow-up (week 72). Among patients with a previous non-response, 12/53 (22.6%; 95% confidence interval [CI] 12.3-36.2%) randomized to triple therapy achieved an SVR compared with 16/52 (30.8%; 95% CI 18.7-45.1%) randomized to the standard of care. Among patients with a previous relapse 22/39 (56.4%; 95% CI 39.6-72.2%) randomized to triple therapy achieved an SVR compared with 23/38 (60.5%; 95% CI 43.4-76.0%) randomized to the standard of care. Undetectable HCV RNA (< 50 IU/mL) at week 12 had a high positive predictive value for SVR. A substantial proportion of non-responders and relapsers to conventional interferon plus ribavirin achieve an SVR when re-treated with peginterferon alfa-2a (40KD) plus ribavirin. Conclusion. Amantadine does not enhance SVR rates in previously treated patients with chronic hepatitis C and cannot be recommended in this setting.

Evolução temporal da proliferação neointimal após implante de dois tipos de stent farmacológico com polímeros biodegradáveis em modelo porcino: avaliação qualitativa por tomografia de coerência óptica sequencial

INTRODUÇÃO: Baseados na hipótese de que a neoíntima encontrada em stents farmacológicos (SFs) com polímeros biodegradáveis aos 28 dias não é a neoíntima definitiva e de que a tomografia de coerência óptica (TCO) é um método eficaz para a avaliação sequencial da neoíntima, objetivamos, neste estudo experimental, comparar os achados da TCO aos 28 dias e aos 90 dias em dois tipos de SF com polímeros biodegradáveis: o stent liberador de sirolimus (Inspiron®, Scitech) e o stent liberador de biolimus A9 (Biomatrix®, Biosensors International). MÉTODOS: No total, 6 porcos não-ateroscleróticos foram submetidos a implante de 6 stents Inspiron® e de 6 stents Biomatrix®. Cada porco recebeu os dois tipos de stent, um em cada artéria coronária (descendente anterior e circunflexa) e após 28 dias e 90 dias foram realizadas avaliações qualitativas intrastent a cada milímetro com TCO. RESULTADOS: A avaliação qualitativa, feita por pareamento milímetro a milímetro intrastent, evidenciou neoíntima heterogênea em 39% aos 28 dias e em 0% aos 90 dias, presença de tecido intraluminal em 18% aos 28 dias e em 0% aos 90 dias, irregularidade luminal em 62% aos 28 dias e em 2% aos 90 dias (P < 0,005). Não houve diferença entre os grupos quanto à qualidade da neoíntima ao longo do tempo (P > 0,05). CONCLUSÕES: Os achados à TCO corroboram a hipótese de que a neoíntima encontrada em SFs com polímeros biodegradáveis aos 28 dias não é a neoíntima definitiva. A evidência experimental mais significativa é a mudança das características da neoíntima observada à TCO sequencial.