Mollicute Anti-Adhesive and Growth Inhibition Properties of the Methanolic Extract of Propolis from the Brazilian Native Bee Melipona quadrifasciata.

Hydroalcoholic propolis extracts from the bee species Melipona quadrifasciata have been shown to possess antimicrobial activity against different mollicute strains, but a methanolic extract (ME) could contain an increased diversity of nonpolar bioactive components with a potentially higher antimicrobial activity. The ME obtained by maceration of the propolis sample was fractionated with solvents of different polarities and then, purified by silica gel column chromatography through biomonitoring of its antimicrobial activity against mollicute strains. Analysis by gas chromatography-mass spectrometry (GC/MS) enabled the identification of compounds using the NIST library. Minimum inhibitory concentrations (MICs) of the samples were determined by broth microdilution. Anti-adhesive assays were performed with Mycoplasma pneumoniae cells. The hexane (MIC=62.5 mg/L) and dichloromethane (MIC=125 mg/L) fractions presented the most promising results against M. pneumoniae. They were fractionated into 74 subfractions, and even the best ones did not show better results (MIC>250 mg/L) than their original fractions, likely due to the loss of terpene compounds that seem to act in synergy. The dichloromethane subfraction FD4 was highlighted in the anti-adhesive assay with an inhibitory activity of 21.6 %. A synergistic effect of the nonpolar compounds in M. quadrifasciata propolis may be responsible for its antibacterial activity, but several purified components can improve its anti-adhesive properties.

Longitudinal analysis of laboratory tests in a population-based study / Análise longitudinal da qualidade dos exames laboratoriais em um estudo de base populacional

ABSTRACT Clinical laboratories must have quality management systems that guarantee the reliability of their results. Furthermore, in longitudinal studies, it is important that the generated data be analyzed over time, as an additional measure of quality control (QC), seeking to identify fluctuations not explainable by biological variables. Applying this strategy to the SHIP-Brazil study was the aim of this work. Thus, we analyzed the results of fasting glucose, post-load glucose and glycated hemoglobin in the participants of the SHIP-Brazil study, from July 2014 to November 2016, in relation to the following aspects: difference of each individual's results from the mean of the month over time, mean and median of results over time, and percentage of results above the reference values over time, with trend line. According to the observed data, in order to guarantee the correct association of laboratory parameters with different health and disease conditions, the fasting blood glucose and post-load glucose measurements in the samples collected during the first months of the study are consistent, despite being performed in two different facilities. This QC strategy has proved very useful, and may even be used by clinical laboratories in their routine, observing result fluctuations of their population in the course of time, being capable of eventually detect bias even before the routine practices of internal and external quality control.

RESUMEN Los laboratorios clínicos deben tener sistemas de gestión de calidad que garanticen la fiabilidad de los resultados. Además, en estudios longitudinales, es importante que los datos generados sean analizados a lo largo del tempo, como medida adicional de control de calidad (QC), buscando identificar fluctuaciones no explicables por variables biológicas. Aplicar esa estrategia al estudio SHIP-Brasil es el objetivo de este trabajo. Analizamos los resultados de glucemia en ayunas, prueba de sobrecarga de glucosa y hemoglobina glucada en los participantes del estudio SHIP-Brasil, en el período de julio de 2014 a noviembre de 2016, acerca de los siguientes aspectos: diferencia de los resultados de cada individuo con respecto a la media del mes con el tiempo; media y mediana de los valores con el tiempo; y porcentaje de resultados por encima de los valores de referencia con el tempo, con línea de tendencia. Según los datos observados, las mediciones de glucemia en ayunas y tras una sobrecarga de glucosa, en las muestras recolectadas en los primeros meses de estudio, resultan consistentes, a pesar de haber sido realizadas en dos laboratorios distintos. Esa estrategia de QC se presentó muy útil y puede, incluso, ser utilizada por laboratorios clínicos en su rutina, observando fluctuaciones de resultados de su población a lo largo del tiempo, detectando, eventualmente, desviaciones aún antes de las prácticas rutinarias de control interno y externo de calidad.

RESUMO Os laboratórios clínicos devem ter sistemas de gestão da qualidade que garantam a fidedignidade dos resultados. Ainda, em estudos longitudinais, é importante que os dados gerados sejam analisados ao longo do tempo, como medida adicional de controle de qualidade (CQ), buscando identificar flutuações não explicáveis por variáveis biológicas. Aplicar essa estratégia ao estudo SHIP-Brasil foi o objetivo deste trabalho. Analisamos os resultados de glicemia em jejum, glicemia pós-sobrecarga e hemoglobina glicada nos participantes do estudo SHIP-Brasil, no período de julho de 2014 a novembro de 2016, em relação aos seguintes aspectos: diferença dos resultados de cada indivíduo em relação à média do mês ao longo do tempo; média e mediana dos valores ao longo do tempo; e porcentagem de resultados acima dos valores de referência ao longo do tempo, com linha de tendência. De acordo com os dados observados, as dosagens de glicemia em jejum e glicemia pós-sobrecarga, nas amostras coletadas nos primeiros meses de estudo, apresentam-se consistentes, mesmo tendo sido realizadas em dois laboratórios distintos. Essa estratégia de CQ mostrou-se bastante útil e pode, inclusive, ser utilizada pelos laboratórios clínicos em sua rotina, observando as flutuações de resultados de sua população ao longo do tempo, detectando, eventualmente, desvios antes mesmo das práticas rotineiras de controle interno e externo da qualidade.

Avaliação da qualidade e da estabilidade de amostras de soro humano para análise e armazenamento em biorrepositório / Evaluation of the quality and the stability of human samples for analysis and storage in biorepository

Objetivo: O Estudo das Condições de Vida e Saúde de Pomerode, SC (SHIP-Brasil), conduzido pela Universidade Regional de Blumenau, visa estratificar os fatores de risco e genéticos eventualmente existentes na gênese das mais variadas doenças. Para tal são necessários exames laboratoriais, antropométricos, genéticos e de imagem para a definição das mesmas e o acompanhamento de sua evolução. Este trabalho teve como objetivo avaliar a qualidade das amostras armazenadas no biorrepositório do estudo, que podem ser utilizadas para pesquisas de biomarcadores no futuro. Métodos: Foram avaliadas amostras de soro em jejum de 56 voluntários. Os analitos estudados foram glicose, colesterol total, creatinina e transaminase glutâmico-pirúvica (ALT), após 1, 2 e 3 anos de armazenamento a -80ºC. Os resultados foram analisados com o Teste t de Student e de acordo com o Erro Permitido Máximo para cada analito. Resultados: Não encontramos evidências de que as amostras do Estudo SHIP-Brasil possam perder a sua qualidade após serem armazenadas por longos períodos. Conclusão: Os Procedimentos Operacionais Padrão (POPs) estabelecidos estão adequados para o processamento e preservação das amostras.

Evaluation of a new equation for LDL-c estimation and prediction of death by cardiovascular related events in a German population-based study cohort.

A simple equation established by Cordova & Cordova (LDL-COR) was developed to provide an improved estimation of LDL-cholesterol in a large Brazilian laboratory database. We evaluated this new equation in a general population cohort in Pomerania, north-eastern Germany (SHIP Study) compared to other existing formulas (Anandaraja, Teerakanchana, Chen, Hattori, Martin, Friedewald and Ahmadi), and its power in the prediction of death by atherosclerosis related events as the primary outcome. Analysis was conducted on a cohort of 4075 individuals considering age, gender, use of lipid lowering therapy and associated co-morbidities such as diabetes, hepatic, kidney and thyroid disease. LDL-COR values had a lower standard deviation compared to the previously published equations: 0.92 versus 1.02, 1.02, 1.03, 1.04, 1.09, 1.10 and 1.74 mmol/L, respectively. All of the factors known to affect the results obtained by the Friedewald's equation (LDL-FW), except fibrate use, were associated with the difference between LDL-COR and LDL-FW (p < .01), with TSH being borderline (p = .06). LDL-COR determined a higher hazard ratio (1.23 versus 1.12, 1.19, 1.21, 1.19, 1.21 and 1.19) for cardiovascular disease related mortality, incident stroke or myocardial infarction compared to the other evaluated formulas, except for Ahmadi's (1.24), and the same adjusted predictive power considering all confounding factors. The proposed simple equation was demonstrated to be suitable for a more precise LDL-c estimation in the studied population. Since LDL-c is a parameter frequently requested by medical laboratories in clinical routine, and will probably remain so, precise methods for its estimation are needed when direct measurement is not available.

Mollicutes/HIV Coinfection and the Development of AIDS: Still Far from a Definitive Response.

Background. Mycoplasmas are known to cause various infections in humans, mainly in the respiratory and urogenital tracts. The different species are usually host-specific and cause diseases in well-defined sites. New species have been isolated, including those from HIV-infected persons. Summary. Its in vitro properties, combined with clinical findings, have led to the hypothesis that these microorganisms may act as cofactors of HIV in AIDS development. Even today this point of view is quite polemic among infectious disease specialists and many aspects remain to be clarified, in contrast to what happens, for instance, with HIV/Mycobacterium tuberculosis coinfection. Dozens of papers have been published covering aspects of Mollicutes/HIV coinfection, but they add little to no information about the putative contribution of Mollicutes to the evolution of AIDS. Very few researchers have devoted their efforts to trying to answer this question, which remains open. In this review, we discuss the evidences that may support this statement in the light of current knowledge in the field of mycoplasmology.