Inhibition of nitric oxide production of activated mice peritoneal macrophages is independent of the Toxoplasma gondii strain

BACKGROUND Toxoplasma gondii causes toxoplasmosis and is controlled by activated macrophages. However, infection of macrophages by tachyzoites induces TGF-β signaling (TGF-s) inhibiting nitric oxide (NO) production. NO inhibition may be a general escape mechanism of distinct T. gondii strains. OBJECTIVES To evaluate in activated macrophages the capacity of T. gondii strains of different virulence and genetics (RH, type I; ME-49, type II; VEG, type III; P-Br, recombinant) to evade the NO microbicidal defense system and determine LC3 loading to the parasitophorous vacuole. METHODS Activated peritoneal macrophages were infected with the different T. gondii strains, NO-production was evaluated by the Griess reagent, and inducible nitric oxide synthase expression, TGF-s, and LC3 localisation assayed by immunofluorescence. FINDINGS Only RH persisted in macrophages, while VEG was more resistant than P-Br and ME-49. All strains induced TGF-s, degradation of inducible nitric oxide synthase, and NO-production inhibition from 2 to 24 h of infection, but only RH sustained these alterations for 48 h. By 24 h of infection, TGF-s lowered in macrophages infected by ME-49, and P-Br, and NO-production recovered, while VEG sustained TGF-s and NO-production inhibition longer. LC3 loading to parasitophorous vacuole was strain-dependent: higher for ME-49, P-Br and VEG, lower for RH. All strains inhibited NO-production, but only RH sustained this effect probably because it persisted in macrophages due to additional evasive mechanisms as lower LC3 loading to parasitophorous vacuole. MAIN CONCLUSIONS These results support that T. gondii can escape the NO microbicidal defense system at the initial phase of the infection, but only the virulent strain sustain this evasion mechanism.

Implantação de um centro na área das violências doméstica e sexual em Campos dos Goytacazes, Rio de Janeiro, 2009-2012 / Implementation of a domestic and sexual violence care center in Campos dos Goytacazes, Rio de Janeiro, Brazil, 2009-2012 / Implementación de un centro en áreas de violencia doméstica y sexual en Campos dos Goytacazes, Río de Janeiro, Brasil, 2009-2012

OBJETIVO: descrever a experiência de implantação do Centro de Atendimento, Pesquisas e Estudos na Área das Violências Doméstica e Sexual (Capevi) em Campos dos Goytacazes-RJ, Brasil, 2009-2012. MÉTODOS: foram convocadas câmaras técnicas entre as secretarias municipais de Saúde e da Família e Assistência Social para implantação do Capevi, visando à reorganização da rede de assistência em urgência e emergência e unificação do instrumento de notificação de violência doméstica e sexual nos sistemas de Notificação de Violências (Sinov) e de Informação de Agravos de Notificação (Sinan). RESULTADOS: após a implantação, cresceram aproximadamente 2000% as notificações no Sinov-Sinan; os tipos de violência com maior incremento de casos no período estudado foram física (de 14 para 435), psicológica/moral (de 4 para 338), tortura (de 3 para 61) e doméstica (de 17 para 200). CONCLUSÃO: o Capevi permitiu o redimensionamento da rede de enfrentamento à violência local e propiciou o aumento quantitativo de notificações.

OBJECTIVE: to describe the experience of implementing a Domestic and Sexual Violence Care and Research Center (Capevi) in the municipality of Campos dos Goytacazes, state of Rio de Janeiro, Brazil, 2009-2012. METHODS: Municipal Health, Family and Social Services Department technical chambers were convened to guide the Center's implantation 2010, with the aim of reorganizing the urgency and emergency care network and unifying the domestic and sexual violence notification instrument on the Violence Notification System (Sinov)and on the Notifiable Diseases Information System (Sinan). RESULTS: after the implementation of the Center notifications on the two Systems increased around 2000%; notifications of violence that incresed the most were physical (from 14 to 435), psychological/moral (from 4 to 338), torture (from 3 to 61) and domestic violence (from 17 to 200). CONCLUSION: the implementation of the Center allowed the city's network to combat violence to be resized and this impacted on increased reporting.

OBJETIVO: describir la experiencia de implementación de un centro de atención, investigación y estudios en el área de violencia doméstica y sexual (Capevi) en el municipio de "Campos dos Goytacazes", estado de Río de Janeiro, Brasil, 2009-2012. MÉTODOS: convocamos cámaras técnicas de las secretarías municipales de salud y familia y, asistencia social para la implantación del Capevi; con el objetivo de reorganizar la red de asistencia a urgencias y emergencias y la unificación del instrumento de notificación de violencia doméstica y sexual en los sistemas de notificación de violencia (Sinov) y Sistema de Información de Enfermedades de Declaración Obligatoria (Sinan). RESULTADOS: después de la implantación, hubo un aumento de hasta 2000% en las notificaciones de los sistemas Sinov-Sinan; los tipos de violencia con mayor incremento de casos en el periodo estudiado fueron la física (de 14 para 453), psicológica/moral (de 4 para 338), tortura (de 3 para 61) y doméstica (de 17 para 200). CONCLUSIONES: el Capevi permitió el redimensionamiento de la red de atención contra la violencia local, propiciando el aumento cuantitativo de notificaciones.

Experimental infection with the Toxoplasma gondii ME-49 strain in the Brazilian BR-1 mini pig is a suitable animal model for human toxoplasmosis

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs' peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.