Mycobacterium marinum infection simulating chromomycosis: a case report

Skins infections caused by Mycobacterium marinum occur only rarely. We report one case of chronic and extensive M. marinum cutaneous infection simulating chromoblastomycosis and review the pertinent literature. A 52-year-old farmer reported a 32-year chronic skin problem on his right lower limb, resulting from contact with cacti. It consisted of skin lesion presenting with dyschromic atrophic center plate and verrucous borders with hematic crusts, extending from the knee anteriorly to the inferior third of the right leg. Mycobacterium marinum infection was detected by histopathological examination of a skin fragment, culture for mycobacteria and genetic mapping of the culture material. The patient was successfully treated with Ethambutol, Rifampicin and Trimethoprim-Sulfamethoxazole. The clinical and histopathological findings of M. marinum infection is nonspecific showing clinical polymorphism and bacilli are rarely evident on histopathological examination. Given these difficulties, it is essential to perform tissue culture in a suspicious case and it is important keep this infection in mind in patients with long-lasting indolent verrucous lesions and a history of exposure to sea water, freshwater, aquaria or fish

Detection of new exemestane metabolites by liquid chromatography interfaced to electrospray-tandem mass spectrometry

Exemestane is an irreversible aromatase inhibitor used for anticancer therapy. Unfortunately, this drug is also misused in sports to avoid some adverse effects caused by steroids administration. For this reason exemestane has been included in World Anti-Doping Agency prohibited list. Usually, doping control laboratories monitor prohibited substances through their metabolites, because parent compounds are readily metabolized. Thus metabolism studies of these substances are very important. Metabolism of exemestane in humans is not clearly reported and this drug is detected indirectly through analysis of its only known metabolite: 17 beta-hydroxyexemestane using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) and gas chromatography coupled to mass spectrometry (GC-MS). This drug is extensively metabolized to several unknown oxidized metabolites. For this purpose LC-MS/MS has been used to propose new urinary exemestane metabolites, mainly oxidized in C6-exomethylene and simultaneously reduced in 17-keto group. Urine samples from four volunteers obtained after administration of a 25 mg dose of exemestane were analyzed separately by LC-MS/MS. Urine samples of each volunteer were hydrolyzed followed by liquid-liquid extraction and injected into a LC-MS/MS system. Three unreported metabolites were detected in all urine samples by LC-MS/MS. The postulated structures of the detected metabolites were based on molecular formulae composition obtained through high accuracy mass determination by liquid chromatography coupled to hybrid quadrupole-time of flight mass spectrometry (LC-QTOF MS) (all mass errors below 2 ppm), electrospray (ESI) product ion spectra and chromatographic behavior. (C) 2011 Elsevier Ltd. All rights reserved

Optimization of an online heart-cutting multidimensional gas chromatography cleanup step for isotopic ratio mass spectrometry and simultaneous quadrupole mass spectrometry measurements of endogenous anabolic steroid in urine

Measuring carbon isotope ratios (CIRs) of urinary analytes represents a cornerstone of doping control analysis and has been particularly optimized for the detection of the misuse of endogenous steroids. Isotope ratio mass spectrometry (IRMS) of appropriate quality, however, necessitates adequate purities of the investigated steroids, which requires extensive pre-analytical sample clean-up steps due to both the natural presence of the target analytes and the high complexity of the matrix. In order to accelerate the sample preparation and increase the automation of the process, the use of multidimensional gas chromatography (MDGC) prior to IRMS experiments, was investigated. A well-established instrumental configuration based on two independent GC ovens and one heart-cutting device was optimized. The first dimension (1D) separation was obtained by a non-polar column which assured high efficiency and good loading capacity, while the second dimension (2D), based on a mid-polar stationary phase, provided good selectivity. A flame ionization detector monitored the 1D, and the 2D was simultaneously recorded by isotope ratio and quadrupole mass spectrometry. The assembled MDGC set-up was applied for measuring testosterone, 5 alpha- and 5 beta-androstanediol, androsterone, and etiocholanolone as target compounds and pregnanediol as endogenous reference compound. The urine sample were pretreated by conventional sample preparation steps comprising solid-phase extraction, hydrolysis, and liquid-liquid extraction. The extract obtained was acetylated and different aliquots were injected into the MDGC system. Two high performance liquid chromatography steps, conventionally adopted prior to CIR measurements, were replaced by the MDGC approach. The obtained values were consistent with the conventional ones. Copyright (C) 2016 John Wiley & Sons, Ltd

Incertezas de medição envolvidas na calibração de vidraria volumétrica de laboratório

3 p. : il.Os laboratórios analíticos nas suas mais diversas áreas de atuação, necessitam garantir a confiabilidade de seus resultados através de procedimentos e técnicas validadas. Como meio de garantir o resultado analítico é necessário que toda a vidraria volumétrica utilizada seja calibrada através de procedimentos que permitam determinar a incerteza de medição. O artigo apresenta o trabalho desenvolvido no Laboratório de Apoio ao Desenvolvimento Tecnológico LADETEC - IQ - UFRJ, na área da calibração de vidraria volumétrica discutindo as variáveis relacionadas as condições ambientais e as possíveis fontes de erro, encontradas durante as operações de calibração. São apresentadas e discutidas as equações utilizadas para o cálculo da estimativa do volume de vidrarias volumétricas e da massa específica da água e do ar. É apresentado um diagrama de causa e efeito para a calibração de vidrarias, descrevendo todas as possíveis fontes de incertezas associadas a cada parâmetro envolvido na estimativa do volume de uma vidraria volumétrica

Different cell death responses induced by Eupomatenoid-5 in MCF-7 and 786-0 tumor cell lines

Natural products remain an important source of new drugs, including anticancer drugs. Recently, our group reported the anticancer activity of eupomatenoid-5 (eup-5), a neolignan isolated from Piper regnellii (Miq.) C. DC. var. regnellii leaves. In vitro studies demonstrated that MCF-7 (breast) and 786-0 (kidney) were among the cancer cell lines most sensitive to eup-5 treatment. The current results demonstrate that mitochondrial membrane depolarization and generation of reactive oxygen species are implicated in eup-5-mediated cytotoxic effects on these cancer cells lines. In MCF-7 cells, eup-5 led to phosphatidylserine externalization and caspase activation, whereas the same did not occur in 786-0 cells. Scanning electron microscopy revealed a reduction of microvilli density, as well as cell morphology alterations. Moreover, treated MCF-7 cells exhibited well-characterized apoptosis alterations, while treated 786-0 cells exhibited characteristics of programmed necroptosis process. These findings support the possibility that different mechanisms may be targeted by eup-5 in cell death response29510261033CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçãosem informação2010/50232-

Implementation and Performance of the Gas Chromatography/Combustion/Isotope Ratio Mass Spectrometry-Based Method for the Confirmatory Analysis of Endogenous Anabolic Steroids during the Rio de Janeiro Olympic and Paralympic Games 2016

Carbon isotope ratio (CIR) confirmation is one of the most complex and delicate analyses in the doping control field, due to the nature of the molecules to be confirmed, normally present in urinary samples as a consequence of an endogenous production. The requirements for method validation established by the World Anti-Doping Agency (WADA) have been pushing the accredited laboratories to improve their methods. The choice of the method is always a cost benefit ratio involving a hard-working and time-consuming analysis and the guarantee of reporting of reliable results. This work presents the method fully validated by the Brazilian Doping Control Laboratory as part of the preparation for the Rio de Janeiro Summer Olympic and Paralympic Games 2016. Sample preparation encompassed solid-phase extraction, liquid-liquid extraction, enzymatic hydrolysis, acetylation, and purification by preparative high-performance liquid chromatography, and analyses were performed by gas chromatography/combustion/isotope ratio mass spectrometry. This proved to be a robust method to CIR confirmation in a big event, as demonstrated by the analysis of 179 samples during the Games 2016, from clearly negative results and adverse findings for testosterone (T) and related substances, boldenone and its metabolite, 19-norandrosterone and formestane. Two atypical findings were also reported for T and metabolites