Ensaio do linfonodo local murino (LLNA) na diferenciação entre a dermatite de contato alérgica e de contato irritante: um estudo da expressão de marcadores de superfície de linfócitos T / Local Lymph node assay in the differentiation between an allergic contact dermatitis and irritation: study of the expression of T lymphocyte surface markers

Introdução: O ensaio do linfonodo local murino (LLNA) foi desenvolvido como uma alternativa aos testes de Buhler e Maximização. O teste é utilizado com o objetivo de identificar substâncias capazes de induzir dermatite de contato e tem como desfecho a quantificação celular nos linfonodos auriculares. Embora recomendado por agências internacionais envolvidas no desenvolvimento de metodologias alternativas, o LLNA ainda necessita de aprimoramento. Objetivo: O objetivo do trabalho foi estudar possíveis diferenças nos padrões de subpopulações linfocitárias entre camundongos tratados com substâncias irritantes e dermosensibilizantes. Método: Os animais foram tratados com os sensibilizantes dinitroclorobenzeno (DNCB) e parafenilenidiamina (PPD), os irritantes lauril sulfato de sódio (LSS) e tritonX-100 (TX-100), por três dias consecutivos no dorso de ambas as orelhas. As subpopulações foram analisadas por citometria de fluxo e possíveis alterações histopatológicas nas orelhas dos animais foram também analisadas. Resultados: Foram observadas diferenças nas células CD4+CD25+ e CD4+CD69+, assim como na proliferação dessas subpopulações. Nenhuma diferença foi vista nos estudos histopatológicos das orelhas dos animais quando tratados com dermosensibilizantes ou irritantes. Conclusões: A fenotipagem de linfócitos T pode ser considerada útil no desenvolvimento de possíveis protocolos de ensaios que visem a diferenciação entre substâncias dermosensibilizantes e irritantes. Além disso, os resultados obtidos podem vir a contribuir com o aumento do conhecimento nesta área e auxiliar na busca por um ensaio in vitro correlato.

Introduction: The Local Lymph Node Assay (LLNA) was developed as an alternative to Buhler and Maximization assays. It is applied to discriminate substances that are able to induce contact dermatitis and the endpoint is cell quantification in mice auricular lymph nodes. Although recommended by international agencies involved in the development of alternative methodologies, LLNA still needs to be improved. Objective: In this context, the goal of this study was to investigate possible differences in lymphocyte subpopulation patterns among mice treated with irritants and dermosensitizers. Method: Animals were treated with sensitizers dinitrochlorobenzene (DNCB) and paraphenylenediamine (PPD) and irritants sodium lauryl sulfate (SLS) and tritonX-100 (TX-100) for 3 days, using dorsum area of both ears. The percentage of different lymphocyte subpopulations were analyzed by flow cytometry. Ears of animals were also evaluated for possible pathological alterations. Results: Differences were observed in CD4+ CD25+ and CD4+ CD69+ cells, as well as in the proliferation of these subpopulations. The histopathological analysis of the ears showed no difference between the treatment with either dermosensitizers or irritants. Conclusions: T lymphocyte phenotyping might still be a useful tool in the development of an assay to differentiate between dermosensitizers and irritants. Moreover, these results may contribute to improving knowledge on this field and helping in the search of a correlate in vitro assay.

Phenotypic profiles and detection of target genes by PCR in isolates from different sources and reference strains, identified as Cronobacter spp. (Enterobacter sakazakii)

Cronobacter, formerly known as Enterobacter sakazakii, is a novel genus of the Enterobacteriaceae family recognized as a cause of high number of fatal cases in neonates, after consuming infant formula. The conventional methods for detecting these organisms are time-consuming and lack sensitivity. The ISO/TS 22964:2006 is the most recently standardized methodology for detecting Cronobacter in powderedinfant formula. This study aimed at confirming the Brazilian isolates previously identified as E. sakazakiias Cronobacter spp. by biochemical assays, and also to compare characteristics of 37 Cronobacter andnon-Cronobacter isolates; and the miniaturized kits and the ISO/TS methodology were evaluated. A conventional PCR protocol targeting dna G was also developed and a previously described gluA targeting protocol was used. The majority of the Brazilian isolates were not confirmed as Cronobacter spp., and the selective enrichment step of ISO/TS methodology was inhibitory to some Cronobacter strains. The ID 32 Ewas the most reliable kit. The PCR protocol targeting gluA showed consistent results with ID 32E and the developed dnaG PCR protocol was 100% sensitive and specific. Thus, the PCR protocols targeting gluA and dnaG might be used to complement the Cronobacter spp. detection or identification after performing the conventional isolation and identification methods.

Aplicação do conceito do erro total na validação do ensaio de potência da vacina oral contra a poliomielite / Use of the total error approach for validating the potency assay for oral poliomyelitis vaccine

Esta pesquisa foi realizada utilizando-se uma abordagem alternativa na validação do ensaio de potênciada vacina oral contra a poliomielite para traçar o perfil deste ensaio, levando-se em conta a grande variabilidade de ensaios biológicos. Foram adotadas duas abordagens para a validação do ensaio: a abordagem Clássica da “International Conference on Harmonization” aplicada aos ensaios biológicos e a abordagem do Conceito do Erro Total. As principais características avaliadas no estudo de validação, por se tratar de um ensaio quantitativo, foram a veracidade, a precisão e a exatidão. Foram ainda avaliadas: a adequação do ensaio aos critérios de aceitação preconizados pelo Food and Drug Administration (EUA) para validação, adotando-se a variação máxima aceita de 20 por cento, a reprodutibilidade do ensaio em uma abordagem prática, com o uso das variâncias entre os resultados de potência de 39 lotes da vacina (do mesmo produtor) obtidos no INCQS e no laboratório produtor, por amostra, para calcular o Coeficiente de Variação geométrico geral. Foi também determinada a Incerteza de Medição do Teste. O ensaio apresentou veracidade e precisão satisfatórias, o que demonstrou sua Exatidão satisfatória para a intenção de uso nas duas abordagens de validação.

Información para la equidad en salud en Chile / Information for health equity in Chile

Objectives. To estimate the magnitude of geographical health inequalities in Chile through key indicators based on data and information that are routinely collected and easily obtained, and to characterize the current situation with respect to the availability, quality, and access to information on health equity that official sources routinely collect. Methods. A conceptual framework proposed by the World Health Organization was used to study health equity in terms of four dimensions: 1) state of health, 2) health determinants, 3) resources for and the supply of health system services, and 4) utilization of health system services. For each of these four dimensions, indicators were selected for which there was available information. The information was aggregated according to geographical and administrative units in the country: communes (342 in Chile), sanitary districts called "Health Services" (28), and regions (13). The aggregated information was analyzed using univariate analysis (distribution characteristics), bivariate analysis (correlations and frequency tables), and tabulation of values for selected indicators for the communes. Results. With respect to the first dimension, state of health, we found an inverse relationship between mortality and average family income in the communes (r = ­0.24; P < 0.001; n = 191 communes). With health determinants, there were important differences among the communes with regard to average household income, years of schooling, literacy, quality of housing, drinking water supply, and the wastewater disposal system. In terms of resources for and the supply of health system services, the municipal governments of the communes with higher average household incomes tended to contribute more funds per beneficiary (r = 0.19; P = 0.013). The financial contributions from the national government were targeted well, but they only partially compensated for the morelimited resources available in poorer communes. With respect to the utilization of health care services per beneficiary in the different sanitary districts, we found some large differences. In terms of the ratio between the highest rate of utilization in any of the districts and the lowest rate in any other district, the ratio for primary-care visits per beneficiary was 2.8, the ratio for emergency-care visits was 3.9, and the ratio for hospitalizations was 2.0. Conclusions. There are important geographical differences in Chile with respect to mortality and other health outcomes, income and environmental conditions, and the financing and utilization of health care services. The information that is collected regularly and is available to characterize the health-related variables frequently has limitations in terms of quality, sustainability, and access. In Chile it would be pointless to focus the greatest efforts on reorganizing the information systems. The existing indicators showing marked inequalities are adequate to support the planning of interventions aimed at making urgently needed improvements in the situation of the worst-off Chileans

Objetivos. Estimar la magnitud de las desigualdades geográficas de salud en Chile mediante indicadores clave basados en datos e información de fácil obtención recolectada de forma rutinaria, y caracterizar la situación actual con respecto a la disponibilidad, calidad y accesibilidad de la información sobre equidad en salud recolectada de forma rutinaria por fuentes oficiales. Métodos. Se usó un marco conceptual propuesto por la Organización Mundial de la Salud para el estudio de la equidad en salud que consta de cuatro dimensiones: I. estado de salud, II. determinantes de la salud, III. recursos y oferta del sistema de salud, y IV. utilización de servicios del sistema de salud. Para cada una de estas dimensiones se hizo una selección de indicadores para los cuales existía información disponible. La información, agregada por unidad geográfica (comuna, Servicio de Salud o región), fue analizada usando los siguientes métodos: análisis univariado (características de las distribuciones); análisis bivariado (correlaciones y tablas de frecuencia); tabulación de valores comunales para indicadores seleccionados. Resultados. Estado de salud: encontramos una relación inversa entre la mortalidad y el ingreso familiar medio en la comuna (r = ­0,24; P < 0,001; n = 191 comunas). Determinantes de la salud: hay importantes variaciones entre las comunas con respecto al ingreso doméstico medio, años de escolarización, analfabetismo, calidad de la vivienda, abastecimiento de agua potable y sistemas de eliminación de aguas residuales. Recursos y oferta del sistema de salud: los gobiernos municipales de las comunas con mayores ingresos domésticos medios tienden a aportar mayores recursos financieros por beneficiario (r = 0,19; P = 0,0130). El aporte del gobierno central, aunque se encuentra bien orientado, solo compensa parcialmente esta situación en beneficio de las comunas pobres. Utilización de servicios de atención de salud: entre los servicios de salud, el uso de los servicios de atención médica en el nivel primario es 2,8 veces mayor en unos que en otros, la atención médica de urgencia 3,9 veces mayor, y los egresos hospitalarios 2 veces más numerosos. Conclusiones. Existen importantes variaciones geográficas en lo que se refiere a la mortalidad y a otros resultados de salud, en ingresos y condiciones ambientales, y en el financiamiento y utilización de los servicios de asistencia sanitaria. La información recolectada regularmente y que se encuentra disponible para caracterizar las variables relacionadas con la salud presenta con cierta frecuencia limitaciones de calidad, sostenibilidad o accesibilidad. En el contexto chileno, sería infructuoso centrar los mayores esfuerzos en la reorganización de los sistemas de información, toda vez que las pruebas ya existentes de marcadas desigualdades son suficientes para apoyar la planificación de intervenciones orientadas a mejorar con urgencia la situación de los chilenos más desprotegidos