Alcohol has been consumed over many centuries, but its connection to criminal activity and accidental fatalities has become a prominent concern in more recent centuries(1). Scientists have developed numerous testing methods to detect alcohol consumption. Numerous studies have recently suggested that zinc has the potential to interfere with the results of these testing methods for drugs of abuse such as enzyme-linked immunosorbent assay (ELISA) and enzyme multiplied immunoassay technique (EMIT) (2, 3). False negatives have been reported from urine testing of drugs such as cocaine, methamphetamine, opiates, and cannabinoids. Nevertheless, minimal research has been conducted concerning zinc’s effect on the adulteration of alcohol metabolite testing. Ethyl glucuronide (EtG) is a promising ethanol metabolite for the confirmation of alcohol consumption. Previous research conducted by Shanna Cawley, a graduate from the Boston University School of Medicine’s Biomedical Forensic Sciences program, has found that zinc sulfate is ineffective at producing conclusive false negative results using two immunochromatographic assay brands in synthetic urine solutions(4). This study uses five different immunoassay brands, five different zinc sources, and two distinct matrices to determine the effectiveness of commercial zinc products as adulterants in drugs of abuse testing. Zinc and EtG solutions were produced at concentrations of 15mg/mL and 750ng/mL, respectively. A positive control, negative control, and two to three experimental trials were conducted for each immunoassay brand and each zinc source resulting in a total of 165 tests. Approximately sixty experimental trials in synthetic urine were invalidated or positive for the presence of EtG (81%) in zinc adulterated EtG solutions. Immunoassay kits produced false positive results when testing human urine from subjects who abstained from alcohol consumption Therefore, preliminary immunoassay screenings for the presence of EtG are not a reliable method for confirming alcohol consumption. Previously researched methods, ELISA and EMIT, and confirmatory methods such as gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) are currently the most robust and reliable techniques for EtG detection in urine.

Il existe un intérêt croissant pour le développement de levures S. cerevisiae œnologiques à rendement abaissé de conversion des sucres en alcool. Nous proposons ici une approche originale basée sur la réorientation du flux carboné vers la voie des pentoses phosphate (VPP). Dans un premier temps, nous avons montré que le flux à travers la VPP est limité par le niveau de réoxydation du NADPH et par la capacité de la voie elle même. Nous avons ensuite mis en évidence le rôle crucial du facteur de transcription Stb5 dans le maintien d'un flux basal à travers la VPP. La surexpression de STB5, couplée à l'introduction d'un système de réoxydation du NADPH, est une stratégie intéressante pour amplifier le flux à travers la VPP. En parallèle, une stratégie d'évolution dirigée basée sur l'adaptation des souches sur gluconate, un hexose mal assimilé et incorporé au niveau de la VPP, a été développée. Des souches évoluées présentant une meilleure assimilation du gluconate ont été obtenues après 70, 180 et 240 générations. En fermentation, ces souches produisent la même quantité d'éthanol que la souche parentale mais présentent des phénotypes complètement nouveaux, en particulier des performances fermentaires accrues, de faibles besoins en azote, une production d'acétate réduite et une forte production de composés aromatiques. L'analyse 13C-flux et transcriptomique d'une souche évoluée ECA5 révèle une amplification de la VPP d'un facteur 1.5 par rapport à la souche parentale EC1118, en lien avec la surexpression de GND1 et TKL1. L'expression de nombreux gènes du métabolisme azoté et de la voie Ehrlich, de l'homéostasie des protons et de la glycolyse est augmentée chez ECA5, alors que les gènes de stress et de la respiration sont globalement réprimés, de façon cohérente avec les phénotypes observés. Outre le développement de nouvelles souches d'intérêt œnologique, ce travail apporte un éclairage nouveau sur le fonctionnement de la VPP et sur ses liens avec le métabolisme central et secondaire / There is an ever-growing interest in the development of S. cerevisiae wine yeast strains with reduced ethanol yield. We proposed a novel approach based on rerouting the carbon flux towards the pentose phosphate (PP) pathway. First, we showed that the flux through the PP pathway is limited both by the absence of a mechanism for reoxidation of NADPH and by the intrinsic capacity of the pathway. We also showed that the transcription factor Stb5 plays a key role in maintaining a basal flux through the PP pathway to meet the requirements for NADPH and biosynthetic precursors. Over-expression of STB5 is a potentially useful strategy for increasing the flux through the PP pathway, provided that an alternative system of reoxidation of NADPH is expressed. In parallel, we investigated an evolutionary engineering strategy based on long-term batch culture on gluconate, a substrate poorly assimilated by S. cerevisiae cells and metabolized by the PP pathway. We selected strains that had evolved a greater gluconate consumption capacity after 70, 180 and 240 generations. During wine fermentation, these evolved strains produced similar amounts of ethanol as the parental strain but displayed completely novel phenotypes, including higher fermentation rates, lower nitrogen requirements, lower levels of acetate production, and enhanced production of aroma compounds. 13C flux analysis and transcripomic analysis of one of these strains, ECA5, showed a greater flux through the PP pathway consistent with the observed increased expression of GND1 and TKL1. The expression of genes associated with nitrogen metabolism, the Ehrlich pathway, proton homeostasis and glycolysis was stronger than in the parental strain, whereas genes involved in stress response and respiration were down-regulated, in agreement with the phenotypes of ECA5. In addition to providing strains with considerable potential for wine making, this work sheds new light on the operation of PP pathway and its links with central and secondary metabolism

Low-fat Cheddar cheese flavor is different from full-fat Cheddar cheese and is not acceptable to many consumers. This 2-part experiment was designed to examine effects adjunct cultures have on low-fat Cheddar cheese flavor as determined through descriptive analysis and consumer feedback. In Part 1, low-fat (5%) Cheddar cheese was produced in duplicate, using 6 combinations of DVS850, LH32, CR540, CRL431, Emfour, and CR319 bacterial cultures. Due to a previously observed positive effect by sodium gluconate on low-fat cheese flavor, each replicate was split into treatments of 0.0% and 0.8% sodium gluconate. Each of these treatments was then split into ripening temperature treatments: 6°C for 21 ± 1 wk; or 6°C for 3 wk, 10°C for 8 wk, and 6°C for 10 wk. Cheese was tasted first by an informal panel. The 4 treatment combinations for the control cheese and the CR540 (a Lactococcus lactis ssp. and Lactobacillus ssp. blend) cheese, along with all culture combinations containing sodium gluconate and ripened only at 6°C, were selected for descriptive analysis. Some statistically significant differences in culture treatment were observed. Sodium gluconate addition had a positive influence on flavor while elevated ripening temperature resulted in undesirable flavor notes. Low-fat (5%) Cheddar cheese with the CR540 adjunct with and without sodium gluconate was evaluated in a consumer taste panel with commercial full-fat (33% fat) and commercial reduced-fat (25% fat) Cheddar cheese. The low-fat cheeses were not significantly different from the commercial reduced-fat, indicating comparable cheese. Part 2 involved making Cheddar-like cheese with non-Cheddar adjunct cultures, using the same process as Part 1. Sodium gluconate was again added but elevated ripening temperature was not included. Each treatment was also divided into a sodium treatment, full salt (2%) and reduced salt (1.5%). After 2 mo of storage at 6°C, cheese was tasted by an informal panel and found to be bitter because of the starter culture used. A culture was added to the second replicate of the experiment to reduce bitterness. This adjunct was found to be somewhat effective in reducing bitterness but not entirely. Descriptive analysis was performed on the high salt level treatments for both replicates. Some difference was observed among cultures and sodium gluconate treatments; however, no acceptable cheese was produced due to bitterness in both replicates. Sodium treatments were not analyzed.

Neisseria meningitidis possède un PTS incomplet. Constitué des composants générales EI et HPr et de deux EIIAs (EIIANtr et EIIAMan), ce système ne permet pas le transport des sucres chez cette bactérie. Cependant, nous avons confirmé que la cascade de phosphorylation (EI HPr EIIANtr) est fonctionnelle et que HPr est aussi phosphorylée sur sa Ser-46 par une HprK/P.Dans l’objectif d’étudier l’effet de HPr sur la virulence de N. meningitidis, nous avons construit un mutant ΔptsH chez N. meningitidis 2C4-3. Le mutant ΔptsH a montré une faible survie chez la souris, une faible production de la capsule, une meilleure adhérence aux cellules épithéliales et un niveau élevé de cellules apoptotiques, par rapport à la souche sauvage. HPr semble intervenir dans la virulence de N. meningitidis en interagissant avec la protéine CrgA. L’interaction HPr/CrgA est plus forte quand HPr est phosphorylée sur sa Ser-46 par HprK/P.N. meningitidis utilise le glucose et le maltose comme seuls sucres. Nous avons identifié une perméase à glucose (GlcP) et une perméase à maltose (MalY), responsables du transport de ces sucres. Une perméase putative à gluconate (GntP) a été également identifiée chez N. meningitidis 2C4-3. Cette perméase n’assure pas le transport du gluconate, dans les conditions testées. La délétion de gntP chez N. meningitidis 2C4-3 induit une meilleure croissance sur glucose et une bonne survie du mutant ΔgntP chez la souris, par rapport à la souche sauvage. La fonction réelle de la perméase GntP chez N. meningitidis reste inconnue et suscite des études ultérieures. / Neisseria meningitidis has an incomplete PTS composed of general proteins EI and HPr and two EIIAs (EIIANtr and EIIAMan). This system does not allow the transport of sugars in this bacterium. However, we confirmed that the phosphorylation cascade (EI HPr EIIANtr) is functional and HPr is also phosphorylated at its Ser-46 by an HprK/P.In order to study the effect of HPr on meningococcal virulence, we constructed a ΔptsH mutant in N. meningitidis 2C4-3. Compared to the wild-type strain, the ΔptsH mutant showed poor survival in mice, low production of capsule, better adherence to epithelial cells and high levels of apoptotic cells. HPr appears to be involved in the virulence of N. meningitidis by interacting with CrgA protein. The HPr/CrgA interaction is stronger when HPr is phosphorylated at its Ser-46 by HprK/P.N. meningitidis uses glucose and maltose as the only sugars. We identified permeases for glucose (GlcP) and maltose (MalY), which catalyze the uptake of these sugars.A putative gluconate permease (GntP) has also been identified in N. meningitidis 2C4-3. Under the conditions tested, this permease did not catalyze the transport of gluconate. Compared to the wild-type strain, deletion of gntP in N. meningitidis 2C4-3 induced faster growth on glucose and a better survival of the mutant in mice. To underst and the function of the GntP permease in N. meningitidis further studies will have to be carried out.

Les matériaux cimentaires sont largement utilisés dans la conception et la construction des sites de stockage de déchets radioactifs. Une des manières d’améliorer leur performance est d’introduire des adjuvants organiques dans la structure. La présence de matière organique dans l’eau porale peut affecter la mobilité des radionucléides : les molécules organiques forment des complexes solubles et peuvent être en compétition avec les radionucléides au niveau des sites de sorption. Ce travail avait pour but de comprendre les mécanismes de telles interactions au niveau moléculaire. Le système modèle a trois composantes. D’abord, des phases C-S-H ont été choisies en tant que modèles du ciment.Ensuite, le gluconate est sélectionné en tant que modèle d’additif organique pour sonder les mécanismes d’interaction à l’échelle moléculaire. Un système plus complexe impliquant un superplastifiant (PCE) a été testé. La troisième espèce, U(VI), est représentative d’un radionucléide de la série des actinides. Le développement de la description des effets de postproduction des espèces organiques pour les applications de stockage des déchets radioactifs était l’objectif principal de ce travail. L’étude des systèmes binaires fournit des données de référence pour l’investigation de systèmes ternaires C-S-H/matière organique/U(VI) plus complexes. Des cinétiques et des isothermes de sorption/désorption pour les espèces sur les C-S-H sont mesurés. En parallèle, des modèles atomiques ont été développés pour les interfaces d’intérêt. Les aspects structuraux, énergétiques et dynamiques des processus de sorption sur les surfaces de ciment sont modélisés par la technique de la dynamique moléculaire. / Cementitious materials are extensively used in the design and construction of radioactive waste repositories. One of the ways to enhance their performance is to introduce organic admixtures into the cement structure. However, the presence of organics in the pore water may affect the radionuclide mobility: organic molecules can form water-soluble complexes and compete for sorption sites. This work was designed to get detailed understanding of the mechanisms of such interactions on the molecular level. The model system has three components. First, pure C-S-H phases with different Ca/Si ratios were chosen as a cement model. Secondly, gluconate (a simple well-described molecule) is selected as a good starting organic additive model to probe the interaction mechanisms on the molecular scale. A more complex system involving polycarboxylate superplasticizer (PCE) was also tested. The third, U (VI), is a representative of the actinide radionuclide series. The development of description of the effects of organics for radioactive waste disposal applications was the primary objective of this work. The study of binary systems provides reference data for the investigation of more complex ternary (C-S-H/organic/U(VI)). The interactions are studied by means of both experimental and computational molecular modelling techniques. Data on sorption and desorption kinetics and isotherms for additives and for U (VI) on C-S-H are acquired in this work. In parallel, atomistic models are developed for the interfaces of interest. Structural, energetic, and dynamic aspects of the sorption processes on surface of cement are quantitatively modeled by molecular dynamics technique.

The purpose of this study was to determine whether chlorhexidine gluconate (0.12%), used as an endodontic irrigating solution would affect the apical seal of three root canal cements. One hundred extracted human single-canal teeth were divided into 9 experimental groups of 10 teeth each, in addition to a positive and negative control group of 5 teeth each. The teeth were decoronated at the level of the cementoenamel junction, accessed, instrumented to a Master Apical File #50, irrigated with either sterile saline, 5.25% NaOCl or 0.12% chlorhexidine gluconate, and dried using paper points. Obturation was accomplished using lateral condensation and one of three endodontic sealers: Roth's 811, AH26, or Sealapex. Post-obturation apical leakage was measured at 270- and 360-day observation periods using the fluid filtration method. Using the mixed-model repeated-measures ANOVA test with Tukey's HSD multiple comparison procedure, the results showed the saline-Sealapex combination had significantly more leakage (p<0.05) than either the Peridex-Sealapex or saline-Roth's combinations at 270 days. No other significant differences were noted between any sealer-irrigant combination at 270 days. The saline-Sealapex combination had significantly more leakage than the saline-Roth's combination at 360 days. No other significant differences were noted at 360 days. Under the conditions of this study, chlorhexidine gluconate irrigant did not adversely affect the apical seal of three root canal cements at 270 and 360 days.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Uma solução de glicerofosfato de sódio, estéril e apirogênica para administração intravenosa, não disponível comercialmente no Brasil, foi desenvolvida e adicionada às nutrições parenterais padronizadas no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo, com o objetivo de fornecer fósforo juntamente com o cálcio. Essas nutrições parenterais foram preparadas adicionando-se 4 mmol/dL de fósforo e 3,712 mmol/dL de cálcio, utilizando-se como fonte as soluções de glicerofosfato de sódio a 1 mmol/mL e gluconato de cálcio a 10%(p/v), respectivamente, e mantidas à temperatura de 5±3ºC por 0, 6, 12 e 24 horas, seguidas de 24 horas à temperatura ambiente (24±3ºC) e posteriormente avaliadas microscopicamente, quando preparadas com emulsão lipídica, com aumento de 400 vezes, e macroscopicamente após centrifugação a 6390 g. As preparações sem emulsão lipídica foram avaliadas macroscopicamente nas mesmas condições e também após a manutenção em temperaturas de 5±3ºC, 24±3ºC e 37°C, durante 96 horas. Em nenhuma formulação, contendo ou não emulsão lipídica, foi verificada a presença do precipitado de fosfato de cálcio. Conclusão: O glicerofosfato de sódio, quando adicionado às nutrições parenterais juntamente com o gluconato de cálcio em quantidades e nas condições avaliadas, não apresentou incompatibilidade in vitro. / A sterile and apyrogenic sodium glycerophosphate solution for intravenous administration, not commercially available in Brazil, was developed and added to standard parenteral nutrition formulations used at the University Hospital, Faculty of Medicine of Ribeirão Preto, University of São Paulo, in order to supply phosphorus together with calcium. These parenteral nutrition formulations were prepared by adding 4 mmol/dL phosphorus and 3.712 mmol/dL calcium using as sources solutions of 1 mmol/mL glycerophosphate and 10% calcium gluconate (w/v) and kept at a temperature (5 + 3oC) for 0, 6, 12 and 24 hours, followed by 24 hours at room temperature of 24 + 3oC. The formulations were then evaluated microscopically, at 400 X magnification, when prepared with a lipid emulsion, and macroscopically after centrifugation at 6390 g. The preparations without a lipid emulsion were evaluated macroscopically under the same conditions and also after being kept at temperatures of 5 + 3oC, 24 + 3oC and 37oC for 96 hours. No calcium phosphate precipitates were observed in any formulation containing or not a lipid emulsion. We conclude that sodium glycerophosphate, when added to parenteral nutrition formulations together with calcium gluconate in the amounts and under the conditions tested here did not present in vitro incompatibility.

Magister Chirurgiae Dentium - MChD (Oral Medicine and Periodontics) / Candida species are frequently isolated from oral mucosal surfaces of healthy individuals and is the most common genus responsible for up to 75% of all candidal infections. The most common problems associated management of oral candidiasis are antifungal drug resistance and side effects Natural medicine is an emerging field and is being explored to overcome drug resistance and to reduce side effects. Gymnemagenin (will be known as Gymnemic acid; GA) is a purified extract from Gymnema sylvestre, a slow growing, perennial, medicinal plant found in Central and Western India, Tropical Africa and Australia is regarded as one of the plants with potent antimicrobial and antifungal activity.

Nitric oxide (NO) has been shown to be lethal for a variety of intracellular pathogens including Leishmania. In murine models, the inducible nitric oxide synthase gene (iNOS) is expressed in activated macrophages and is involved in the synthesis of NO. Because the role of NO and iNOS in human leishmaniasis was less clear, we examined the expression of the iNOS gene in human macrophages infected with Leishmania in vitro and in biopsy tissue from subjects with cutaneous leishmaniasis. / Pentavalent antimony (Sb5+) in the form of Pentostam(TM) or Glucantime(TM) is still the treatment of choice despite its toxicity. Aldara(TM) (5% imiquimod) is an immune-response modifying agent that has been approved by the Food and Drug Administration in the USA for treating genital warts caused by papillomaviruses. We conducted an open-label, prospective study of combined Glucantime(TM) + Aldara(TM) therapy in subjects with CL who had previously failed a complete course of Glucantime(TM) treatment at regular doses. (Abstract shortened by UMI.)