Desenvolvimento de metodologia analítica por HS-SPME-GC-MS para avaliação da presença de metabólitos resultantes da decomposição microbiana da água de lastro de biodiesel

O biodiesel é um combustível menos poluente que os de origem fóssil (petróleo e derivados), produzido a partir de fontes renováveis, como óleos vegetais e gorduras animais. Quimicamente são ésteres metílico ou etílico de óleos e gorduras obtido por meio da reação de transesterificação de triglicerídeos com álcool metanol ou etanol. Apesar das inúmeras vantagens do biodiesel, o contato com o ar e com a água durante a estocagem pode afetar a sua qualidade; por ser altamente higroscópico. Dependendo das condições de transporte e armazenamento, há proliferação de micro-organismos envolvidos na degradação do combustível, na água de lastro. A água de lastro possui em sua composição uma série de compostos derivados do óleo do qual foi originada. Com intuito de sanar ou reduzir esse problema, determinados fungos são utilizados para conter esta proliferação. Um dos fungos amplamente utilizados é o Pseudallescheria boydii. A avaliação da composição da água de lastro em termos de compostos resultantes dos processos de degradação envolvendo fungos e micro-organismos é fundamental para a caracterização do sistema e eficiência dos fungos. Para a identificação de compostos residuais resultantes da degradação microbiana da água de lastro na presença e ausência de fungos, foi desenvolvida uma metodologia analítica baseada em Cromatografia a Gás acoplada à Espectrometria de Massas que permitisse identificação por tentativa destes compostos. A análise por tentativa é utilizada quando não há padrões disponíveis para os compostos de interesse e consiste em comparar o perfil de fragmentação do espectro de massa gerado com a biblioteca NIST. Uma das técnicas de preparo de amostra mais empregadas para esse tipo de matriz é a SPME (Solid Phase Microextraction). A partir desta problemática, o objetivo do trabalho foi desenvolver metodologia analítica por SPME para análise por cromatografia a gás acoplada a espectrômetro de massas (GC-MS) para compostos resultantes da decomposição microbiana de água de lastro de biodiesel. As melhores condições para a SPME foram estabelecidas possibilitando a comparação entre o perfil de compostos das amostras na ausência e presença do fungo. Na comparação do perfil cromatográfico das amostras controle, os ésteres de cadeia longa são detectados já na primeira semana. Na presença de fungos a ocorrência de ésteres de cadeia longa pode ser detectada somente a partir da quarta semana.Biodiesel is a less polluting fuel than fossil fuels (petroleum and derivatives), produced from renewable sources such as vegetable oils and animal fats. Chemically they are methyl or ethyl esters of oils and fats obtained by a transesterification reaction of triglycerides with methanol or ethanol. Despite the many advantages of biodiesel, contact with air and water during storage can affect its quality; being highly hygroscopic. Depending on the conditions of transport and storage, proliferation of microorganisms involved in fuel degradation in free water, may occur. Free water has a series of oil-derived compounds in its composition, springing from its original source. In order to remedy or reduce this problem, certain fungi are used. A widely used fungus is Pseudallescheria boydii. The evaluation of free water composition, in terms of compounds resulting from degradation processes involving fungi and microorganisms is essential for the characterization of the system and ascertainment of the efficiency of the fungi. – For identification of residual compounds, resulting from microbial degradation of free water in presence and absence of fungi, an analytical methodology based on Gas-Chromatography coupled to Mass Spectrometry that allows tentative identification of these compounds was developed. Tentative identification is applied when no standards are available for the interest compounds and consists in comparing the fragmentation profile of the generated mass spectrum with the NIST library. One of the most commonly used sample preparation techniques for this type of matrix is SPME (Solid Phase Microextraction). From this problematic, the objective of this work was to develop SPME analytical methodology for analysis by Gas-Chromatography coupled to Mass Spectrometry (GC-MS) for compounds resulting from microbial decomposition of biodiesel free water. The best conditions for SPME were established allowing the comparison between the compound profile of the samples in the absence and presence of the fungus. Comparing the chromatographic profile of control samples, long chain esters are detected as early as the first week. In the presence of fungi, the occurrence of long chain esters can be detected only from the fourth week

Comparison of Value Set Based on DCE and/or TTO Data: Scoring for EQ-5D-5L Health States in Japan

AbstractBackgroundThe valuation study of the five-level version of the EuroQol five-dimensional questionnaire (EQ-5D-5L) involved composite time trade-off (cTTO) and a discrete choice experiment (DCE). The DCE scores must be anchored to the quality-of-life scale from 0 (death) to 1 (full health). Nevertheless, the characteristics of the statistical methods used for converting the EQ-5D-5L DCE results by using TTO information are not yet clearly known.ObjectivesTo present the Japanese DCE value set of the EQ-5D-5L and compare three methods for converting latent DCE values.MethodsThe survey sampled the general population at five locations in Japan. 1098 respondents were stratified by age and sex. To obtain and compare the value sets of the EQ-5D-5L, the cTTO and DCE data were analyzed by a linear mixed model and conditional logit, respectively. The DCE scores were converted to the quality-of-life scale by anchoring to the worst state using cTTO, mapping DCE onto cTTO, and a hybrid model.ResultsThe data from 1026 respondents were analyzed. All the coefficients in the cTTO and DCE value sets were consistent throughout all the analyses. Compared with the cTTO algorithm, the mapping and hybrid methods yielded very similar scoring coefficients. The hybrid model results, however, produced a lower root mean square error and fewer health states with errors exceeding 0.05 than did the other models. The DCE anchored to the worst state overestimated the cTTO scores of almost all the health states.ConclusionsJapanese value sets based on DCE were demonstrated. On comparing the observed cTTO scores, we found that the hybrid model was slightly superior to the simpler methods, including the TTO model

Associations among baseline variables, treatment-related factors and health-related quality of life 2 years after breast cancer surgery

Provision of social support and rehabilitation for patients with physical, mental, and functional problems after cancer treatment is important for long-term health-related quality of life (HRQOL). Effective use of human and financial healthcare resources requires identification of patients requiring rehabilitation. The objectives of the current study were to clarify the patterns of physical and psychosocial recovery over time, to evaluate the associations among baseline variables, treatment-related factors and HRQOL at 6 months, 1, and 2 years after breast cancer surgery, and to identify the significant factors predicting HRQOL at each point. A multicenter longitudinal study was performed to evaluate physical conditions, anxiety, depression, and HRQOL at 1 month (baseline), 6 months, 1, and 2 years after surgery in 196 patients (mean age: 53.3 years old) with early breast cancer and no postoperative recurrence. Physical conditions were evaluated using a patient-reported symptom checklist. HRQOL was rated using the functional assessment of cancer treatment scalegeneral (FACT-G) and the breast cancer subscale (FACTB). Anxiety and depression were rated using the hospital anxiety and depression scale (HADS). More than 50% of patients had local problems of "tightness", "arm weakness." and "arm lymphedema", and systemic problems of "reduced energy, fatigue, and general weakness" postoperatively. The HRQOL score significantly improved 1 year after surgery, and scores for physical, emotional and functional well-being also increased with time, whereas the score for social well-being was the highest at baseline and decreased with time. Depression and anxiety significantly improved with time. Concomitant disease, marital status, and the presence of a partner, anxiety and depression at baseline, pathological lymph node involvement, and adjuvant intravenous chemotherapy were significant factors predicting FACT-G scores at 6 months, 1, and 2 years after surgery. Depression at baseline was a strong predictor of HRQOL up to 2 years after surgery. These results suggest that physical rehabilitation is required for tightness and lymphedema to improve long-term postoperative physical function. A further study of psychosocial interventions is required to improve depression and social well-being after breast cancer surgery

Opposing role of condensin hinge against replication protein A in mitosis and interphase through promoting DNA annealing

Condensin is required for chromosome dynamics and diverse DNA metabolism. How condensin works, however, is not well understood. Condensin contains two structural maintenance of chromosomes (SMC) subunits with the terminal globular domains connected to coiled-coil that is interrupted by the central hinge. Heterotrimeric non-SMC subunits regulate SMC. We identified a novel fission yeast SMC hinge mutant, cut14-Y1, which displayed defects in DNA damage repair and chromosome segregation. It contains an amino acid substitution at a conserved hinge residue of Cut14/SMC2, resulting in diminished DNA binding and annealing. A replication protein A mutant, ssb1-418, greatly alleviated the repair and mitotic defects of cut14-Y1. Ssb1 protein formed nucleolar foci in cut14-Y1 cells, but the number of foci was diminished in cut14-Y1 ssb1-418 double mutants. Consistent with the above results, Ssb1 protein bound to single-strand DNA was removed by condensin or the SMC dimer through DNA reannealing in vitro. Similarly, RNA hybridized to DNA may be removed by the SMC dimer. Thus, condensin may wind up DNA strands to unload chromosomal components after DNA repair and prior to mitosis. We show that 16 suppressor mutations of cut14-Y1 were all mapped within the hinge domain, which surrounded the original L543 mutation site