Dissertation submitted in partial compliance with the requirements of the Master's Degree in Technology: Homoeopathy, Technikon Natal, 2002. / The purpose of this study was to determine the effect of herbal extracts of certain Labiatiae species (Rosmannus officinalis, Salvia officinalis and Thymus vulgaris) in the inhibition of in vitro growth of Candida albicans as compared to ethanol as a control and nystatin in terms of the disc diffusion test. / M

The glycolytic enzyme enolase is an abundant, immunodominant antigen of the fungal pathogen Candida albicans. A C. albicans enolase cDNA was used to study the genetic organization and expression of enolase genes. Experimental results were consistent with a model predicting four enolase genes per diploid genome. Enolase steady state mRNA levels increased during logarithmic growth, but were unaffected by pH, growth form or carbon source.

Thesis (M.Sc.D.)--Boston University School of Graduate Dentistry. Dept. of Orthodontics. 1970. / The present investigation was undertaken to determine an extra-skeletal reference plane and its reliability. The results of this study revealed that it seems possible for an orthodontist to establish an extra-skeletal reference plane, if a specially designed analyzer is used. It has been shown that by using this devise in his determination of a base landmark, an orthodontist can have a high degree of reliability within himself. It also appears likely that the reference plane obtained in this manner by one orthodontist is reproducible, to a high degree of reliability by other orthodontists. A remarkably high reproducibility, as obtained if a group determination of a reference base was done. To the orthodontist, it means that there may be a way of accurately producing a reliable reference plane for static cephalocentric analysis. This could be used to standardize communication among orthodontists.[TRUNCATED]

C. albicans est un microorganisme opportuniste présent chez plus de 60% de la population. La pathogénicité de C. albicans est contrôlée par le système immunitaire de l’hôte, grâce entre autres aux peptides antimicrobiens (PAMs). Ces PAMs peuvent être de différentes origines. Notre étude a pour but d’évaluer l’effet d’un PAM d’amphibiens, la dermaseptine-S1 (DS1), sur la croissance, la transformation et l’expression de certains gènes de virulence de C. albicans. C. albicans (ATCC-SC5314) a été cultivé en présence ou en absence de la DS1 à 10, 50, et 100 μg/ml, pendant 24 h puis la croissance a été évaluée par un teste de numération cellulaire. Le passage de la forme levure à la forme hyphe a été évalué par microscopie optique et électronique à balayage après culture en présence ou en absence de la DS1 pendant 3 et 6 h à 37°C et ce en présence de 10% de sérum de veau foetal. L’effet de la DS1 sur la formation de biofilm a été évalué par un test MTT et par microscopie électronique à balayage. Enfin, l’expression de certains gènes protéase aspartique sécrétée (Saps 1, 2, 3, 9, 10 et Hwp1) a été évaluée par qRT-PCR. Nous avons montré que la DS1 réduit de façon significative la croissance de C. albicans. La DS1 inhibe le passage de C. albicans de sa forme blastospore à la forme hyphe, ainsi que la formation du biofilm. L’effet de la DS1 sur C. albicans pourrait passer par un contrôle de l’expression de plusieurs gènes impliqués dans la sécrétion d’enzyme protéolytique. En effet, la DS1 diminue de façon significative l’expression des gènes Saps 1, 2, 3, 9 et 10 ainsi que du gène Hwp1. L’ensemble de nos travaux démontre que la DS1 peut contrôler la pathogénicité de C. albicans. / Candida is an opportunistic microorganism present in more than 60% of the population. The pathogenicity of Candida is controlled by the host immune system, such as antimicrobial peptides (AMPs). These PAMs can be of different origins. Our study is to evaluate the effect of an amphibian-isolated antimicrobial peptide, dermaseptin-S1 (DS1) on the growth, transformation and the expression of certain C. albicans virulence genes. C. albicans (ATCC-SC5314) was cultured in the presence or the absence of DS1 at 10, 50, and 100 μg / ml for 24 h and then the growth of yeast was assessed by cell count. The transformation of yeast was evaluated at 3 and 6 h of culture under conditions conducive to transformation (37°C in the presence of 10% of serum) by optical and scanning electron microscopy. Biofilm formation was assessed using a collagen scaffold, after incubation for 30 min at 30°C, with MTT assay and scanning electron microscopy. Finally, the expression of some secreted aspartic protease genes (Saps 1, 2, 3, 9, 10 and Hwp1) was evaluated by qRT-PCR. The results showed that DS1 significantly reduces the growth of C. albicans. This effect is proportional to the concentration of the peptide. This peptide inhibits the passage of C. albicans from the blastospore to the hypha form. The effect of DS1 on C. albicans involves controlling the expression of several genes. Indeed, DS1 significantly decreases the expression of Sap 1, 2, 3, 9 and 10 genes as well as Hwp1 gene. All of our work demonstrates that DS1 can control the pathogenicity of C. albicans.

Candida albicans est une levure pathogène opportuniste responsable d’infections appelées candidoses. Ce type d’infection est particulièrement grave chez les personnes ayant un système immunitaire affaibli. Toutefois, peu de traitements sont disponibles et une augmentation des résistances font que les infections fongiques sont un problème de santé publique majeur. Au sein de l’hôte, cette levure doit s’adapter à l’environnement parfois hostile des différentes niches. Afin de persister chez l’être humain, ce micro-organisme doit être en mesure de percevoir les changements environnementaux et adapter son métabolisme en conséquence. L’adaptation aux nutriments disponibles ainsi que de la teneur en oxygène sont deux éléments importants pour le développement dans les différentes niches. La compréhension de l’adaptation de cette levure au sein de l’hôte pourra permettre l’identification de nouvelles cibles thérapeutiques. Nous avons effectué une analyse métabolomique à différents temps d’exposition à l’hypoxie dans le but de découvrir pour la première fois la réponse métabolomique de C. albicans lorsque cette levure est confrontée à une déplétion en oxygène. Nous avons observé un remodelage métabolique important en particulier concernant la glycolyse, la voie des pentoses phosphates et la paroi cellulaire. Le lipidome de C. albicans est également fortement affecté par l’hypoxie, en particulier les composants de la membrane plasmique tels que les phospholipides, les sphingolipides et l’ergostérol. Les données transcriptomiques corroborent les résultats observés lors de l’étude métabolomique. Enfin, nous avons testé la sensibilité de C. albicans à différents stress afin de confirmer l’altération physiologique provoquée par l’hypoxie comme suggérée par l’analyse du métabolome. Nous avons également réalisé un criblage de banques de mutants afin d’identifier des régulateurs de la flexibilité métabolique hypoxique. Nous avons identifié le rôle de Snf5, une sous-unité du complexe de remodelage de la chromatine SWI/SNF, permettant de coordonner la disponibilité en nutriments et en oxygène avec la réponse transcriptionnelle pour permettre l’adaptation aux changements environnementaux. Snf5 est nécessaire pour la réponse transcriptionnelle relative au commensalisme et aussi aux traits de virulence de C. albicans. Nous avons confirmé ces observations par des tests de colonisation intestinale dans un modèle murin et le mutant snf5 s’est révélé incapable de persister dans le tractus digestif des souris. Le mutant snf5 présente également une atténuation des traits de pathogénicité : il ne peut former des hyphes, ne cause pas de dommage aux cellules de l’hôte et il est avirulent dans un modèle d’infection des larves de Galleria mellonella. L’étude des interactions génétiques a permis d’identifier un lien entre SNF5 et le gène de l’adénylate cyclase CYR1. Ceci suggère que Snf5 et Cyr1 appartiennent à la même voie de signalisation afin de permettre la flexibilité métabolique hypoxique. Les travaux de cette thèse ont permis de connaitre la réponse métabolomique hypoxique chez C. albicans et apportent des connaissances fondamentales sur cette levure. Cette compréhension pourra par la suite servir à cibler les voies métaboliques importantes pour la survie au sein de l’hôte et ainsi permettre le développement de nouveaux antifongiques. Nous avons également identifié le rôle de Snf5 pour la flexibilité métabolique hypoxique. L’absence de Snf5 provoque un défaut majeur de virulence chez C. albicans, ceci renforce l’idée de cibler les voies métaboliques pour lutter contre cette levure. Ainsi Snf5 ou un autre membre du complexe SWI/SNF pourrait être une cible thérapeutique pour le développement de nouveaux traitements antifongiques.

L’intestin humain contient une variété de microbes commensaux qui sont représentés par divers organismes appartenant aux trois domaines de la vie où les Eukarya sont essentiellement représentés par le règne des champignons. La levure commensale et opportuniste Candida albicans a été identifiée comme étant le champignon le plus commun dans l’intestin des humains sains. Des études récentes soutiennent que malgré leur faible abondance les levures du genre Candida peuvent altérer l'équilibre du microbiote et conduire à des dysbioses ou des pathologies récurrentes comme la maladie de Crohn et les colites ulcéreuses. Il a été démontré que le microbiote commensal joue un rôle essentiel dans la protection de l’intestin contre la colonisation par des bactéries pathogènes et des pathobiontes. Cependant, jusqu'à présent, on ignore si la prolifération ou la pathogénicité de C. albicans peuvent être contrôlées par d'autres microbiotes fécaux. Dans cette étude, nous avons démontré que le métabolome microbien de l'intestin humain exerce une activité antifongique contre C. albicans et d’autres levures qui résident au niveau intestinal. Ces métabolites inhibent plusieurs traits de virulence de C. albicans incluant la filamentation et l'invasion des cellules humaines. De plus, un crible génétique chez C. albicans a suggéré que TOR est la cible moléculaire de la ou des molécules antifongiques du métabolome microbien de l'intestin humain. / The human gut contains a variety of commensal microbes which are composed of diverse organisms that belong to all three domains of life with Eukarya primarily represented by fungi. The commensal / opportunistic yeast Candida albicans has been reported as the most common fungus in the gut of healthy humans. Recent evidences support that, this small fraction can alter the microbiota equilibrium leading to dysbiosis and diseases like inflammatory bowel diseases. It has been demonstrated that commensal microbiota plays a critical role in the protection of the gut against colonization by other bacterial pathogens and pathobionts. However, so far, whether C. albicans overgrowth or pathogenicity are controlled by other fecal microbiota is not known. In this study, we showed that the human microbial gut metabolome (GM) exerts an antifungal activity against different intestinal-resident yeasts including hyphal growth and the invasion of human enterocytes of C. albicans. Furthermore, a genetic screen in C. albicans suggested that TOR is the molecular target of the antifungal molecule(s) of the GM.

Thesis (M. Sc.)--Université Laval, 1999. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M. Sc.)--Université Laval, 1999. / Includes bibliographical references.

Orientadores: Altair Antoninha Del Bel Cury, Tatiana Pereira Cenci / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-15T11:15:55Z (GMT). No. of bitstreams: 1 Fernandes_FredericoSilvadeFreitas_M.pdf: 733963 bytes, checksum: a861bd550ab439f138ef016317f27638 (MD5) Previous issue date: 2010 / Resumo: Biofilme de Candida spp formado na superfície de próteses removíveis é considerado o principal fator etiológico da candidose, a qual é a infecção oral fúngica mais prevalente em humanos. Em pacientes com comprometimento motor, o uso de limpadores químicos é indicado para o controle desse biofilme, entretanto, pouco se conhece sobre o efeito desses agentes sobre o biofilme de Candidas não-albicans. Adicionalmente, a literatura é escassa de estudos avaliando a formação de biofilme de Candida sobre novos materiais para base de próteses. Assim, o objetivo desse estudo foi avaliar o efeito de limpadores químicos sobre o biofilme mono e multi-espécie de Candida formado sobre a superfície de materiais para confecção de próteses removíveis. Foram confeccionados espécimes de resina de polimetilmetacrilato (PMMA) e resina poliamida, os quais, após a padronização da rugosidade de superfície (0,34 ± 0,02 µm), foram submetidos à avaliação da energia livre de superfície (ELS) ou à formação de biofilme. Biofilme de Candida albicans e/ou Candida glabrata foi formado por 72 h, sendo os espécimes, previamente, submetidos à formação da película adquirida. Após o período de formação do biofilme, os espécimes foram submetidos aos tratamentos, segundo o tempo recomendado por cada fabricante: limpador químico enzimático (3 min); limpador químico sem enzimas (5 min); e hipoclorito de sódio (NaOCl) a 0,5% (10 min). A água destilada e deionizada foi utilizada como controle. Após os tratamentos, os espécimes foram sonicados (7W por 30s) em solução salina, para remoção das células aderidas. Essa solução foi serialmente diluída em solução salina e semeada em CHROMagar® Candida. O número de células viáveis de Candida foi expresso em unidades formadoras de colônia (UFC)/mm2. Os dados da ELS e ângulo de contato foram submetidos a ANOVA um fator, enquanto que os dados de células viáveis de Candida foram submetidos a ANOVA três fatores, seguido do teste de Tukey-Kramer. Todos os biofilmes avaliados apresentaram maior crescimento na resina de poliamida (p<0,0001), entretanto, essa resina apresentou um menor valor de ELS quando comparada à resina de PMMA. Os limpadores químicos, contendo ou não enzimas, reduziram significantemente os níveis de Candida, sem haver diferença estatística entre eles (p=0,9999). Entretanto, o NaOCl a 0,5% foi mais eficaz, na medida em que resultou na ausência de células viáveis. Em todas as situações avaliadas, a C. glabrata apresentou maiores valores de células viáveis do que a C. albicans (p=0,0002). Nas condições desse estudo, conclui-se que a resina de poliamida possibilitou uma maior proliferação de Candida; e os limpadores químicos comerciais foram eficazes na redução dos níveis de Candida spp, mas apenas a solução de hipoclorito de sódio a 0,5% resultou na ausência de células viáveis na superfícies dos materiais testados / Resumo: Os limpadores químicos de prótese têm sido bastante indicados para o controle do biofilme formado sobre próteses removíveis de pacientes com comprometimento motor. Apesar de estudos prévios terem mostrado que uma única imersão nesses agentes é capaz de reduzir os níveis de Candida albicans do biofilme formado sobre próteses removíveis, pouco se sabe sobre o efeito do uso diário desses limpadores sobre o biofilme residual de Candida. Assim, o objetivo desse estudo foi avaliar a eficácia do uso diário de um limpador químico enzimático sobre o biofilme de C. albicans formado sobre a superfície de materiais para confecção de próteses removíveis; bem como a atividade enzimática das células de Candida desse biofilme após exposições diárias a esse limpador de prótese. Foram confeccionados espécimes de resina de polimetilmetacrilato (PMMA) e resina de poliamida, nos quais foi realizada, inicialmente, a padronização da rugosidade de superfície (0,34 ± 0,02 ?m). Após a formação da película adquirida, os espécimes foram divididos aleatoriamente em 12 grupos (n=9) para formação do biofilme de C. albicans por 72 horas. Após esse período, os espécimes foram tratados por 1, 4 ou 7 dias, sendo realizado um tratamento por dia, com um limpador químico enzimático (Polident 3 Minutes) ou com água destilada (controle negativo). Após os respectivos períodos de tratamento, os microrganismos remanescentes foram removidos da superfície dos espécimes por meio de ondas ultra-sônicas (7W por 30s). Em seguida, as unidades formadoras de colônia (UFC) foram calculadas e a atividade enzimática das células remanescentes foi avaliada. Os dados foram submetidos à ANOVA um fator ou dois fatores, seguido do teste de Tukey-Kramer. O biofilme de Candida albicans formado sobre a resina de poliamida apresentou maiores níveis de Candida e uma maior atividade fosfolipásica que o biofilme formado sobre a resina de PMMA (p<0,001). O limpador químico enzimático reduziu significantemente os níveis de Candida albicans em todos os períodos avaliados (p<0,001), entretanto os níveis desse microrganismo aumentaram com o tempo, sendo observada diferença estatisticamente significante entre os períodos avaliados (p<0,001). As exposições diárias a esse limpador químico aumentaram a virulência das células de Candida, no que diz respeito à atividade fosfolipásica. Nas condições desse estudo, conclui-se que o uso diário do limpador químico enzimático não foi capaz de impedir a proliferação de Candida albicans no biofilme residual, apesar de ter interferido no crescimento desse biofilme. / Abstract: Candida denture biofilm is considered the the primary aetiological agent for the development of oral candidosis, which is the most common fungal oral infection in humans. Although, for patients with limited motor capacity, chemical cleansing with immersion in denture cleansers has been shown to be effective in controlling Candida biofilm accumulation, limited data is available on the effect of those cleansing agents on other Candida species biofilms. Additionally, few studies have examined the development of Candida biofims on novel denture materials. This study evaluated the efficacy of denture cleansers on C. albicans and C. glabrata single and dual-species biofilms formed on novel denture base materials. Specimens of polymethylmetacrylate resin (PMMA) and polyamide resin were prepared and had their surface roughness standardized (0.34 ± 0.02 µm). Part of the specimens had their surface free energy measured and the other specimens were submitted to the biofilm assays. C. albicans and/or C. glabrata biofilm was formed for 72 hours on saliva-coated specimens. On the 3rd day, specimens were treated with an enzymatic cleanser, denture cleanser or 0.5% sodium hypochlorite (NaOCl) solution by soaking for, 3, 5 and 10 min, respectively. Water was used as negative control. After treatment, adhered cells were detached from the acrylic resin surface by ultrassonic waves at 7 watts for 30 seconds in phosphate buffered saline solution (PBS). This solution was serially diluted in PBS and plated on CHROMagar® Candida. Candida viable cell were expressed in colony forming units per surface area (CFU/mm2). Data of surface free energy and contact angle were analyzed by one-way ANOVA, and data of Candida species were analyzed by three way-ANOVA followed by Tukey-Kramer test. All tested biofilms displayed significantly higher growth on polyamide thermoplastic resin (p<0.0001), which presented the lowest SFE. Denture cleansers significantly decreased Candida spp levels, with no statistical difference between them (p=0.9999); however, 0.5% NaOCl solution was more effective, since, after treatment, no viable cell was observed. Candida glabrata revealed significantly higher CFU counts when compared to Candida albicans under all experimental conditions (p=0.0002). Our study has shown that polyamide resins may present a convenient substratum for microbial colonization. Although denture cleansers reduced Candida levels, sodium hypochlorite should be preferred as it was efficient to eliminate Candida cells from the tested materials / Abstract: Chemical cleansing with immersion in denture cleansers has been indicated for denture biofilm control in patients with limited motor capacity. Although previous studies have shown that a single immersion in those agents is able to substantially reduce Candida albicans biofilm levels, the effect of the routine use of denture cleansers on the Candida residual biofilm is poorly understood. This study evaluated the efficacy of daily use of an enzymatic denture cleanser on C. albicans biofilm formed on denture base materials; and the enzymatic activities of Candida biofilm cells after daily exposure to this cleanser agent. Polymethyl methacrylate (PMMA) and polyamide resins specimens were prepared (n=54), and their surface roughness was standardized (0.34 ±0.02 ?m). Saliva-coated specimens were randomly divided by lottery into 12 groups (n=9) for biofilm assay. C. albicans biofilm was formed for 72 hours, and then specimens were treated for 1, 4 or 7 days, once a day, with an enzymatic cleanser (Polident 3 Minutes), or distilled water (negative control). Remaining adherent microorganisms were removed from the treated specimens by ultrasonic waves at 7 watts for 30 seconds, and then colony-forming units (CFU) were calculated and remaining cells enzymatic activities were determined. Data were analyzed by 1-way or 2-way ANOVA followed by the Tukey-Kramer test. C. albicans biofilm formed on polyamide resin showed significantly higher Candida levels and phospholipase activity (p<0.001) than biofilm formed on PMMA resin. The enzymatic cleanser significantly reduced C. albicans levels in all evaluated periods (p<0.001); however, the number of this microorganism increased with time, showing statistical difference among the treatment periods (p<0.001). The daily exposure to the denture cleanser increased Candida cells virulence, with regard to phospholipase activity. Our study has shown that the enzymatic cleanser daily use did not prevent C. albicans proliferation in residual biofilm; however, this agent reduced this fungus rate of growth. / Mestrado / Protese Dental / Mestre em Clínica Odontológica

Call number: LD2668 .T4 1964 B62 / Master of Science