Improved Canine and Human Visceral Leishmaniasis Immunodiagnosis Using Combinations of Synthetic Peptides in Enzyme-Linked Immunosorbent Assay

Visceral leishmaniasis is endemic in many areas of tropical and subtropical America where it constitutes a significant public health problem. It is usually diagnosed by enzyme-linked immunosorbent assays (ELISA) using crude Leishmania antigens, but a variety of other immunological methods may also be applied. Although these approaches are useful, historically their sensitivity and specificity have often been compromised by the use of complex mixtures of antigens. In this context, the use of combinations of purified, well-characterized antigens appears preferable and may yield better results. In the present study, combinations of peptides derived from the previously described Leishmania diagnostic antigens A2, NH, LACK and K39 were used in ELISA against sera from 106 dogs and 44 human patients. Improved sensitivities and specificities, close to 100%, for both sera of patients and dogs was observed for ELISA using some combinations of the peptides, including the detection of VL in dogs with low anti-Leishmania antibody titers and asymptomatic infection. So, the use of combinations of B cell predicted synthetic peptides derived from antigens A2, NH, LACK and K39 may provide an alternative for improved sensitivities and specificities for immunodiagnostic assays of VL

The Cysteine-Rich Protein Thimet Oligopeptidase as a Model of the Structural Requirements for S-glutathiolation and Oxidative Oligomerization

Thimet oligopeptidase (EP24.15) is a cysteine-rich metallopeptidase containing fifteen Cys residues and no intra-protein disulfide bonds. Previous work on this enzyme revealed that the oxidative oligomerization of EP24.15 is triggered by S-glutathiolation at physiological GSSG levels (10–50 µM) via a mechanism based on thiol-disulfide exchange. In the present work, our aim was to identify EP24.15 Cys residues that are prone to S-glutathiolation and to determine which structural features in the cysteinyl bulk are responsible for the formation of mixed disulfides through the reaction with GSSG and, in this particular case, the Cys residues within EP24.15 that favor either S-glutathiolation or inter-protein thiol-disulfide exchange. These studies were conducted by in silico structural analyses and simulations as well as site-specific mutation. S-glutathiolation was determined by mass spectrometric analyses and western blotting with anti-glutathione antibody. The results indicated that the stabilization of a thiolate sulfhydryl and the solvent accessibility of the cysteines are necessary for S-thiolation. The Solvent Access Surface analysis of the Cys residues prone to glutathione modification showed that the S-glutathiolated Cys residues are located inside pockets where the sulfur atom comes into contact with the solvent and that the positively charged amino acids are directed toward these Cys residues. The simulation of a covalent glutathione docking onto the same Cys residues allowed for perfect glutathione posing. A mutation of the Arg residue 263 that forms a saline bridge to the Cys residue 175 significantly decreased the overall S-glutathiolation and oligomerization of EP24.15. The present results show for the first time the structural requirements for protein S-glutathiolation by GSSG and are consistent with our previous hypothesis that EP24.15 oligomerization is dependent on the electron transfer from specific protonated Cys residues of one molecule to previously S-glutathionylated Cys residues of another one

Chlorhexidine with or without alcohol against biofilm formation: efficacy, adverse events and taste preference

Abstract In recent years, different chlorhexidine formulations have been tested, including an alcohol-free alternative, but the effect of this solution on early biofilm formation is not clear. A crossover, randomized, double-blind clinical trial was conducted to evaluate the effect of two chlorhexidine solutions against supra- and subgingival biofilm formation (NCT#02656251). Thirty-five participants were randomized and asked to rinse twice daily with 15 ml of an alcohol-containing 0.12% chlorhexidine solution, an alcohol-free 0.12% chlorhexidine solution, or placebo. The study was conducted in three experimental periods of 4 days each, with a 10-day washout between the periods. All the experimental periods followed the same protocol, except that the solutions were switched. Biofilm distribution was evaluated every 24 hours by the Plaque-Free Zone Index, during 96 hours. Adverse events were self-reported and sensory evaluation was performed using a hedonic scale. Compared to the placebo, the chlorhexidine solutions resulted in a significantly higher number of surfaces free of plaque over 96 hours (p < 0.01), and were able to prevent subgingival biofilm formation (p < 0.01). The alcohol-free chlorhexidine solution was associated with a lower incidence of adverse events, compared with alcohol-containing chlorhexidine (p < 0.05); it also received better sensory evaluation and acceptance by trial participants, compared with the alcohol-containing chlorhexidine (p = 0.007), and had a similar inhibitory effect on the formation of supra- and subgingival biofilms

Phytoplankton dynamics in a highly eutrophic estuary in tropical Brazil

The port of Recife in northeastern Brazil is an important ecological and social area but little is known about its environmental quality. Observations, sampling and measurements of phytoplankton were performed during spring and neap tides in the dry (January-February, 2005) and rainy (June, 2005) seasons to assess the environmental quality of the port of Recife. The area had chlorophyll-a concentrations ranging from 3.30 to 54.40 mg m-3, the highest values occurring at low tide during the dry season. A total of 129 species were identified: 53 of them in the dry season and 97 in the rainy season. Diatoms were the most diverse group and comprised 75.47% of the phytoplankton collected in the dry season and 60.82% of those collected in the rainy season. The dry season was characterized by Coscinodiscus sp. and Helicotheca tamesis; the rainy season by Oscillatoria sp. and Coscinodiscus centralis. Species diversity indices varied from 1.06 to 3.74 bits cel-1. Low indices were related to the dominance of Helicotheca tamesis, Coscinodiscus centralis, Coscinodiscus sp. and Aulacoseira granulata. Seasonal forcing, rather than the estuarine flux, determined the phytoplankton community structure. The area is exposed to seasonally varying negative impacts due to metropolitan degradation and the high level of eutrophication.Para avaliar a qualidade ambiental Porto do Recife (Nordeste do Brasil), uma área ecológica e socialmente importante mas pouco investigada, observações, medições e amostragem do fitoplâncton foram realizadas durante as marés de sizígia e quadratura no período de estiagem (janeiro-fevereiro / 2005) e chuvoso (junho/2005). A área apresentou concentração de clorofila a variando de 3,30 a 54,40 mg m-3, com maiores valores registrados durante a maré baixa no período de estiagem. Um total de 129 espécies foi identificada com 53 espécies durante o período de estiagem e 97 espécies no chuvoso. As diatomáceas formaram o grupo mais diverso, com 75,47% no período de estiagem e 60,82% no chuvoso. Coscinodiscus sp. e Helicotheca tamesis caracterizaram o período de estiagem e Oscillatoria sp. e Coscinodiscus centralis o período chuvoso. A diversidade de espécie variou de 1,06 a 3,74 bits cel-1 . Os baixos índices foram relacionados com a dominância de Helicotheca tamesis, Coscinodiscus centralis, Coscinodiscus sp. e Aulacoseira granulata. A sazonalidade determinou a estrutura da comunidade fitoplanctônica, em vez do fluxo estuarino. A área é exposta sazonalmente a vários impactos negativos, devido à degradação metropolitana e ao elevado nível de eutrofização

Características químicas dos mingaus desidratados de arroz e soja

O presente trabalho teve por objetivo caracterizar quimicamente os mingaus desidratados de arroz e soja nas respectivas proporções 100:0, 90:10, 80:20, 70:30, 60:40 e 50:50% e verificar a possibilidade do uso de alguns destes mingaus como ingredientes na formulação de alimentos infantis com base nos resultados discutidos. O processo utilizado para sua obtenção foi decorticagem dos grãos de soja, branqueamento, desintegração e homogeneização do arroz e soja, assim como secagem por atomização. A caracterização química foi realizada através das seguintes determinações: composição centesimal, composição de minerais e de aminoácidos, e atividade do inibidor de tripsina. Com o aumento das proporções de soja (de 0 a 50%), os mingaus desidratados tiveram aumento nos teores de proteína, extrato etéreo, cinzas, fibra bruta, fósforo, cálcio, potássio, magnésio, ferro, cobre, zinco, selênio, manganês e molibdênio, porém diminuição nos teores de carboidrato, sódio e cromo. Contudo, os valores de cobalto não foram alterados. Os teores de potássio, magnésio, ferro e cobre foram altos e os teores de cromo, selênio, manganês e molibdênio foram suficientes em 100g de mingaus desidratados de arroz e soja com 40 e 50% de soja, atendendo 100% das recomendações da RDA para lactentes, podendo estes formulados para alimentos infantis. Houve uma melhoria nos escores de aminoácidos essenciais com as crescentes proporções de soja, exceto para os aminoácidos sulfurados. A composição em aminoácidos foi adequada e comparável ao padrão da FAO/WHO para lactentes, exceto para lisina e metionina. Em relação a lisina, os mingaus desidratados com 30, 40 e 50% de soja mostraram os maiores escores para pré-escolares, não havendo deficiência para escolares. Por outro lado, a metionina não foi deficiente para pré-escolares e escolares. A atividade residual do inibidor de tripsina não foi observada nos mingaus desidratados estudados