Development and comparative evaluation of two antigen detection tests for Visceral Leishmaniasis.

BACKGROUND: Visceral leishmaniasis (VL) can be fatal without timely diagnosis and treatment. Treatment efficacies vary due to drug resistance, drug toxicity and co-morbidities. It is important to monitor treatment responsiveness to confirm cure and curtail relapse. Currently, microscopy of spleen, bone marrow or lymph node biopsies is the only definitive method to evaluate cure. A less invasive test for treatment success is a high priority for VL management. METHODS: In this study, we describe the development of a capture ELISA based on detecting Leishmania donovani antigens in urine samples and comparison with the Leishmania Antigen ELISA, also developed for the same purpose. Both were developed as prototype kits and tested on patient urine samples from Sudan, Ethiopia, Bangladesh and Brazil, along with appropriate control samples from endemic and non-endemic regions. Sensitivity and specificity were assessed based on accurate detection of patients compared to control samples. One-Way ANOVA was used to assess the discrimination capacity of the tests and Cohen's kappa was used to assess their correlation. RESULTS: The Leishmania Antigen Detect ELISA demonstrated >90% sensitivity on VL patient samples from Sudan, Bangladesh and Ethiopia and 88% on samples from Brazil. The Leishmania Antigen ELISA was comparable in performance except for lower sensitivity on Sudanese samples. Both were highly specific. To confirm utility in monitoring treatment, urine samples were collected from VL patients at days 0, 30 and 180 post-treatment. For the Leishmania Antigen Detect ELISA, positivity was high at day 0 at 95%, falling to 21% at day 30. At day 180, all samples were negative, corresponding well with clinical cure. A similar trend was also seen for the Leishmania Antigen ELISA albeit; with lower positivity of 91% at Day 0 and more patients, remaining positive at Days 30 and 180. DISCUSSION: The Leishmania Antigen Detect and the Leishmania Antigen ELISAs are standardized, user- friendly, quantitative and direct tests to detect Leishmania during acute VL as well as to monitor parasite clearance during treatment. They are a clear improvement over existing options. CONCLUSION: The ELISAs provide a non-invasive method to detect parasite antigens during acute infection and monitor its clearance upon cure, filling an unmet need in VL management. Further refinement of the tests with more samples from endemic regions will define their utility in monitoring treatment.

Anti-parasite therapy drives changes in human visceral leishmaniasis-associated inflammatory balance.

Visceral leishmaniasis (VL) remains a major public health problem worldwide. Cytokine balance is thought to play a critical role in the development of this disease. Here, we perform a prospective exploratory study addressing whether simultaneous assessment of circulating levels of different lipid mediators and cytokines could highlight specific pathways involved with VL pathogenesis. VL patients displayed substantial increases in serum levels of Prostaglandin F2α (PGF2α), Leukotriene B4 (LTB4), Resolvin D1 (RvD1), IL-1ß, IL-6, IL-8, IL-10, IL-12p70 and TNF-α compared with uninfected endemic control group, while exhibiting decreased levels of TGF-ß1. Hierarchical cluster analysis of the prospective changes in the expression level of theses parameters upon anti-Leishmania treatment initiation revealed that the inflammatory profile observed in active disease gradually changed over time and was generally reversed at day 30 of therapy. Furthermore, not only the individual concentrations of most of the inflammatory biomarkers changed upon treatment, but the correlations between those and several biochemical parameters used to characterize VL disease activity were also modified over time. These results demonstrate that an inflammatory imbalance hallmarks active VL disease and open perspective for manipulation of these pathways in future studies examining a potential host-directed therapy against VL.

Elaboração de biscoitos de chocolate com substituição parcial da farinha de trigo por polvilho azedo e farinha de albedo de laranja / Chocolate biscuits preparation with partial substitution of wheat flour by fermented cassava starch and orange albedo flour

Os consumidores estão cada vez mais exigentes e preocupados com a saúde e, por consequência, exigindo produtos naturais que deverão ser seguros e promover qualidade de vida. O presente trabalho teve o objetivo de desenvolver biscoitos com farinhas mistas de polvilho azedo e farinha de albedo de laranja, utilizando um planejamento fatorial 2³, com variáveis independentes: concentração de polvilho azedo, açúcar e farinha de albedo de laranja, e variáveis respostas: as características sensoriais: aparência, aroma, textura e sabor, além da impressão global, utilizando escala hedônica estruturada de nove pontos. Os biscoitos foram avaliados também quanto às características físico-químicas. As médias obtidas para as características sensoriais estiveram na faixa que vai de cinco (não gostei nem desgostei) até sete (gostei moderadamente), não apresentando diferença significativa (P≤0,05) para todas as características sensoriais avaliadas. Dessa maneira, foi possível escolher a formulação ideal obtida de acordo com os parâmetros nutricionais, por não existir diferença sensorial significativa, que foi 35 por cento de polvilho azedo, 100 por cento de açúcar e 7,5 por cento de farinha de albedo de laranja. No teste sensorial comparativo com os biscoitos comercial, padrão e otimizado, as características sensoriais aparência, sabor e textura apresentaram diferença significativa (P≤0,05). As características físico-químicas apresentaram resultados correspondentes aos padrões estabelecidos pela legislação brasileira, e o teor de fibra bruta do biscoito otimizado foi de 3,08 por cento, assim, pode ser classificado como biscoito "fonte de fibras".

The consumers are increasingly demanding and worried about health and, consequently, requiring natural products that should be safe and that promote life quality. The present research had the objective to develop biscuits with mixed flour of fermented cassava starch and orange albedo flour, using a 2³ factorial experimental. The independent variables were: concentration of fermented cassava starch, sugar and orange albedo flour, and the responses variables were the sensory characteristic: appearance, aroma, texture and flavor, in addition to overall impression, using a nine point hedonic structure scale. The biscuits were also evaluated for physic-chemical properties. The average for sensory characteristic were in the range of five (not liked nor disliked) up to seven (like moderately), and it didn't show significant difference (P≤0.05) for all sensory characteristics assessed. Thus, it was possible to choose the optimal formulation obtained according with the nutritional parameters, because there isn't sensory difference significant, which was: 35 percent of fermented cassava starch, 100 percent of sugar and 7.5 percent of orange albedo flour. In comparison with the commercial biscuits test, standard and optimized, sensory appearance, flavor and texture showed significant differences (P≤0.05). The results correspond to standards set by Brazilian legislation, and crude fiber content of the biscuit was optimized at 3.08 percent, thus, can be classified as biscuit "source of fiber".

Avaliação do consumo alimentar e do perfil lipídico de mulheres na menopausa / Evaluation of alimentar consumption and lipidic profile of menopause women

Este estudo tem como objetivo avaliar o consumo alimentar e o perfil lipídico em mulheres na menopausa. Foi realizadocom 72 pacientes, atendidas no laboratório de análises clínica do HU/UFS através do SUS, com idade entre 45 e 60 anos, menopausadas,submetidas à avaliação do consumo alimentar e do perfil lipídico. O perfil nutricional foi obtido através de questionário de freqüência de consumo alimentar (QFCA) validado pelo National Cholesterol Education Program, o qual classifica as pacientes em estágios I, II e III. O perfil lipídico foi aferido através de dosagens séricas, pelo analisador bioquímico automatizado - Dimension RXL (DadeBehring), sendo que as frações do colesterol LDL e VLDL foram obtidas pela equação de Friedewald. Em relação ao consumo alimentar, cerca de 20,8% das pacientes foram classificadas no estágio II. Quanto ao perfil lipídico, 31% apresentaram níveis baixo deHDLc, também foi observado níveis elevados de colesterol total e LDLc, em 22,2% e 25% da população estudada, respectivamente. Portanto, os resultados reforçam a necessidade de conscientização dessa população sobre a importância do perfil nutricional adequado para a melhora da qualidade de vida, reduzindo, assim, a incidência de doenças cardiovasculares.

The objective of this study was evaluated the food consumption and lipid profile in women on menopause. This study evaluated 72 patients’ users by SUS, of analyses clinical laboratory at Sergipe’s Federal University Hospital. The patients had age between 45 and 60 years old, all on menopause. They were submitted of food consumption questionnaire (FCQ) validated by National Cholesterol Education Program, and their lipid profile were also valuated. The lipidic profile was mensured by serial dosage, using a biochemical analyzer- Dimension RXL (Dade Behring), and the cholesterol LDL and VLDL fractions obtained thought Friedewald. In relation to food consumption, about 20,8% of the patients were classified in period of training II. About the lipidic profile, 31% of patients presented low levels of HDLc, was also observed high levels of LDLc fraction and total cholesterol, 25% and 22% respectively. Therefore, this results shows the necessity of nutritional education in the evaluated population for the improvement of life’s quality, reducing the incidence of cardiovascular disease.