Orientador: Ricardo de Souza Cavalcante / Resumo: Introdução. A candidemia tem-se tornado cada vez mais frequente nos hospitais e apresentado elevada mortalidade. Conhecer a epidemiologia e os fatores prognósticos permite melhor manejo clínico desta infecção. Diante disto, este estudo teve por objetivo avaliar a incidência da candidemia em um hospital de ensino durante os últimos anos, a mortalidade, a prevalência das espécies causadoras da infecção, bem como os fatores prognósticos. Métodos. Este foi um estudo de coorte não concorrente, com pacientes atendidos no Hospital das Clínicas da Faculdade de Medicina de Botucatu (HCFMB), que apresentaram diagnóstico de candidemia durante sua internação, entre os anos de 2012 e 2019. Os dados clínicos foram obtidos a partir do prontuário médico. Para a avaliação dos fatores associados ao prognóstico, em análise univariada e múltipla, foi utilizado o modelo de Regressão Proporcional de Cox. Foram consideradas significativas as variáveis com valores de p inferiores a 0,05. Resultados. Ocorreram 314 episódios de candidemia em 288 pacientes. A taxa de incidência foi de 1,66/1000 admissões. A espécie mais prevalente foi Candida albicans (38,8%), seguida de complexo C. parapsilosis (20,4%), C. tropicalis (15,3%), C. glabrata (10,8%) e C. krusei (2,5%). Na população pediátrica houve predomínio de complexo C. parapsilosis (p<0,0001) e C. glabrata nos adultos (p=0,002). A taxa de mortalidade em 30 dias foi de 43,4%, sendo maior em adultos do que pacientes pediátricos (19,3% vs 54,0%; p<0,01)... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction. Candidemia has become increasingly frequent in hospitals and has shown high mortality. Knowing the epidemiology and prognostic factors allows better clinical management of this infection. In view of this, this study aimed to evaluate the incidence of candidemia in a teaching hospital during the past few years, mortality, the prevalence of species causing the infection, as well as prognostic factors. Methods. Non-concurrent cohort study was carried out, with inpatients at the Hospital das Clínicas, Faculdade de Medicina de Botucatu (HCFMB), who presented a diagnosis of candidemia during their hospitalization, between the years 2012 and 2019. The clinical data were obtained from the medical records. For the assessment of factors associated with prognosis, in univariate and multiple analysis, the Cox Proportional Regression model was used. Variables with p values below 0.05 were considered significant. Results. There were 314 episodes of candidemia in 288 patients. The incidence rate was 1.66 / 1000 admissions. The most prevalent species was Candida albicans (38.8%), followed by C. parapsilosis complex (20.4%), C. tropicalis (15.3%), C. glabrata (10.8%) and C. krusei (2.5%). In the pediatric population, there was a predominance of the C. parapsilosis complex (p <0.0001) and C. glabrata in adults (p=0.002). The 30-day mortality rate was 43.4%, being higher in adults than pediatric patients (19.3% vs 54.0%; p<0.01). Independent factors of mortality were liver cirrhos... (Complete abstract click electronic access below) / Mestre

Carbon dots (CD) have emerged as the new eye-catching theranostic nanomaterials due to their distinctive features, including tunable emission, facile surface modification, high biocompatibility and low cytotoxicity. These distinguishing features allow full customizations of CD according to the needs of various studies. Of note, they have been widely employed as nano-vehicles with live-tracking systems in many biological applications to deliver medicine with low bioavailability to targeted sites. Candida albicans, a commonly seen commensal fungus accounts for life-threatening infections in humans, is the leading cause of oral candidiasis. Yet, the efficacy of the "gold standard" Amphotericin B (AmB) has been limited due to poor water solubility and dose-dependent cytotoxicity. In addition, the interactions of CD with Candida cells/biofilms and human epithelial tissues have not been fully investigated, and very limited studies have been done on CD-based antifungal drugs delivery for topical administration. Herein, AmB-conjugated guanylated CD (CD-Gu + -AmB) tackling oral fungal infections were synthesized and possessed potent antifungal/anti-biofilm effects against C. albicans. Moreover, CD-Gu + -AmB exhibit low cytotoxicity to primary human oral keratinocytes and can selectively accumulate in the cell nuclei. Above all, the first evidence of studying the penetration and exfoliation profiles of CD in a three- dimensional organotypic human oral epithelial tissue model was provided, and the accumulation of CD-Gu + -AmB in the epithelial tissue can form a 'shielding' layer on oral epithelia against C. albicans. This study demonstrates that CD-Gu + -AmB may serve as a promising antifungal agent for tackling C. albicans and Candida-induced oral candidiasis through fast epithelial penetration, extra-/intra-cellular embedding and gradual exfoliation

Occidiofungin is a novel glycolipopeptide, synthesized and secreted by Burkholderia contaminans MS14, demonstrating broad-spectrum antifungal activity and potential for successful clinical applications. Its mechanism of action has not yet been determined but is known to exhibit fungicidal activity via the induction of apoptosis in a manner unique from that of currently approved antifungals. As an early investigation into occidiofungin’s mechanism of action, we aimed to identify environmental and cellular factors that significantly alter the susceptibility of the model organism, Saccharomyces cerevisiae. To that end, we have demonstrated that occidiofungin’s bioactivity requires active cellular growth, that new protein synthesis is necessary to adequately respond to occidiofungin exposure, and that alterations in transcriptional regulation in response to glucose and phosphate deprivation have synergistic and antagonist consequences, respectively, on occidiofungin’s effectiveness. Together, this data provides a foundation on which occidiofungin’s mechanism of action can be illuminated.

Fungal infections have a significant impact on the world population, with estimates of over 1.6 million deaths a year. One contributing factor is the increasing number of fungi resistant to the current clinical treatments, including the last approved family of antifungal compounds introduced into the market over a decade ago. This is driving the search for new antifungals with different biological targets. A new potential antifungal occidiofungin, is a cyclic glycolipopeptide isolated from the soil bacterium Burkholderia contaminans MS14 with a broad spectrum of activity against both human and plant pathogens. Kill kinetics demonstrated that this compound is fungicidal and activates the cell wall integrity pathway at suboptimal dosing as determined by Mkc1 MAPK phosphorylation studies. As three of the four classes of currently available antifungals target ergosterol or ergosterol biosynthesis, the bioactivity of occidiofungin was assayed in the presence of ergosterol containing DOPC vesicles and was shown to retain antifungal properties. Occidiofungin was also found to have a similar activity profile against the S. cerevisiae -1,3-glucan synthesis mutant, indicating that it does not share a target with the fourth class of antifungals. Stability testing showed occidiofungin retained in vitro potency in the presence of human serum, across a broad range of pH and temperature conditions, and was resistant to gastric proteases. Based on cell morphology, occidiofungin did not target a specific stage of the yeast cell cycle, however cells were smaller in size and acquired ‘dancing bodies’, both properties of apoptosis. This was confirmed with data showing concentration dependent increases in DNA fragmentation, reactive oxygen species, and extracellular localization of phosphatidylserine. In addition to these findings, cells deleted for the yeast caspase gene exhibit a 2old resistance to occidiofungin. While SEM showed no morphological differences between treated and untreated cells, TEM did identify a thinning of the cell wall and inclusion bodies in cells treated with occidiofungin. As a stable fungicidal compound that induces apoptosis in yeast, occidiofungin has a great potential to become a new candidate drug for clinical use in treating fungal infections, including those resistant to current antifungals.

Ongoing antibiotic drug discovery is vital as antimicrobial resistance continues to be a significant issue faced in the clinic. Natural products have long been a highly productive source to mine for new antimicrobials. While it has been challenging to discover new and unique antimicrobial natural products, numerous drugs have been derived from studying how natural products function as secondary metabolites. Previous studies suggested that screening natural product extract fraction libraries for antimicrobials can be more productive than screening crude extracts alone. These studies from large industrial enterprises are generally not directly portable to an academic setting due to significant infrastructure costs. We developed a screening platform consisting of low pressure reversed-phase chromatographic separation of methanolic extracts of bacteria and fungi to generate a prefractionated natural product library. This platform is suitable for academic labs to screen for antimicrobial compounds. A large growth inhibitor screen against multiple pathogens and lab strains of microbes was conducted to assess the validity of the advantages of screening fraction libraries versus crude extract libraries and to search for potential new drug-like compounds. Hits were investigated for reproducibility, isolated, and purified. One compound was discovered in an antifungal screen which may be a novel lipopeptide. / Thesis / Master of Science (MSc) / To combat the growing antibiotic resistance crisis, new strategies for drug discovery must be investigated and implemented. Natural products from bacteria and fungi have long been a source of critically important drugs. Prior research suggests that looking for vital natural products in fraction libraries can be more productive than screening crude extracts. Here the development and assessment of a fractionation library suitable for an academic lab's ingrained limitations are described. Assessing the library indicates an increased hit rate on screening fractions compared to crude extract. Furthermore, pursuing these hits may have revealed a novel antifungal lipopeptide.

Yes / There are many human oral antimicrobial peptides responsible for playing important roles including maintenance, repairing of oral tissues (hard or soft) and defense against oral microbes. In this review we have highlighted the biochemistry, physiology and proteomics of human oral histatin peptides, secreted from parotid and submandibular salivary glands in human. The significance of these peptides includes capability for ionic binding that can kill fungal Candida albicans. They have histidine rich amino acid sequences (7–12 family members; corresponding to residues 12–24, 13–24, 12–25, 13–25, 5–11, and 5–12, respectively) for Histatin-3. However, Histatin-3 can be synthesized proteolytically from histatin 5 or 6. Due to their fungicidal response and high biocompatibility (little or no toxicity), these peptides can be considered as therapeutic agents with most probable applications for example, artificial saliva for denture wearers and salivary gland dysfunction conditions. The objectives of current article are to explore the human histatin peptides for its types, chemical and biological aspects. In addition, the potential for therapeutic bio-dental applications has been elaborated. / King Saud University

No / Herein we report a library of new ruthenium(II) complexes which incorporate a range of functionalised β -ketoiminate ligands. The complexes undergo an unusual reduction from Ru(III) to Ru(II), and consequently incorporate carbonyl ligands from the 2-ethoxyethanol solvent, forming ruthenium dicarbonyl complexes. In order to address the potential applications of these complexes, we have screened the library against a range of tumour cell lines, however, all compounds exhibit low cellular activity and this is tentatively assigned to the decomposition of the compounds in aqueous media. Studies to establish the antifungal and antibacterial potential of these complexes was addressed and show increased growth inhibitions for C. neoformans and S. aureus species. / The authors would like to thank the universities of Leeds, Huddersfield and Bradford for internal financial support.

The polyene macrolide antibiotic natamycin (Antibiotic A-5283) is commonly used to retard the growth of surface molds on various cheese varieties. Natamycin is commonly applied to the surface of cheese by dipping or spraying, using an aqueous dispersion containing 200 to 300 ppm of the additive. The large molecular weight of natamycin, 666 g/mol, and conjugated double bond structure causes it to be extremely insoluble in water and most food grade solvents. The inability to apply natamycin in true solution creates void non-treated areas on the food surface. These non-treated areas promote the growth of fungal organisms. A water soluble N-alkyl semisynthetic derivative of natamycin was synthesized by the Michael addition reaction of the parent with a N-substituted malemide. A comparative study investigating the effectiveness of the semisynthetic derivative of natamycin and the parent antibiotic in suppressing mold growth on one month aged shredded Cheddar cheese modified atmosphere packaged (MAP) was performed. A 20 ppm natamycin treatment effectively suppressed visible mold growth (<104 CFU/g) in MAP samples for up to 30 days after opening. The 20 ppm semisynthetic derivative performed similarly to the 10 ppm natamycin treatment in retarding mold growth. Visible mold growth did not occur for these treatments in MAP samples until 20 days after opening. Analysis of storage conditions revealed that an outgrowth of mold in shredded cheese occurred in MAP packages stored longer than 15 days. This bloom in mold growth was attributed to the degradation of natamycin and the semisynthetic derivative throughout storage. The stability and degradation of natamycin and the derivative were monitored throughout the study. Antibiotic concentration on the cheese surface was quantified by molecular absorption spectrometry. Results from this study showed, heavily contaminated samples caused the rate and loss of natamycin and the derivative to increase. Antibiotic concentration decreased at a similar rate in MAP and open package conditions. Natamycin and derivative were found to have similar degradation properties. / Master of Science

Xia Lixin. / "August 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 210-224). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Les moisissures sont responsables de contaminations sur les produits de BVP et induisent des pertes économiques conséquentes. Dans ce contexte, les cultures bioprotectrices représentent un intérêt croissant comme alternative aux conservateurs chimiques. L’objectif de la première partie de cette étude a été d’évaluer in vitro et in situ l’activité antifongique de bactéries lactiques et propioniques contre cinq moisissures contaminants isolées de produits de BVP. Les bactéries les plus actives pendant les tests in vitro ont été testées in situ par pulvérisation de surface. Sur le milieu WFH, les isolats bactériens les plus actifs correspondent aux espèces Lactobacillus plantarum, reuteri et au groupe buchneri. Les souches plus actives après ces tests ont été testées par inclusion dans la recette du pain au lait et différentes souches ont montré un effet retard en particulier la souche Leuconostoccitreum qui semble retarder la croissance de Penicillium corylophilum après 10 jours. Dans la deuxième partie de cette étude, les surnageants de cultures actifs sont analysés pour identifier les composés antifongiques grâce à différentes traitements et différentes méthode telle que l’HPLC et la spectrométrie de masse. Les résultats suggèrent que les acides organiques jouent un rôle prépondérant dans l’activité antifongique et ont montré que les composés antifongiques retrouvés correspondaient aux acides lactique, acétique et propionique, à l’éthanol et au peroxyde d’hydrogène, ainsi que d’autres composés mais à plus faible échelle. Sur ces résultats, différentes combinaisons des composés identifiés ont été testées pour leur effet sur la germination et la croissance radiale de P. corylophilum et E. repens. Certaines de ces combinaisons ont montré les mêmes effets que le surnageant actif ce qui confirme l’implication des molécules identifiées dans l’activité. Les résultats suggèrent que l’acide acétique est responsable de la totalité de l’activité antifongique observée sur P. corylophilum et qu’il joue un rôle important dans l’inhibition de E. repens. La souche bactérienne sélectionnée pourrait représenter une possibilité de culture bioprotectrice pour les produits de BVP. / Molds are responsible for the spoilage of bakery products and thus, cause substantial economic losses. In this context, bioprotective cultures represent a growing interest as an alternative to chemical preservatives. The aims of the first part of this study was to evaluate the in vitro and in situ antifungal activity of lactic acid bacteria (LAB) and propionibacteria against five moulds species isolated from bakery products. The most inhibitorybacteria found during the in vitro test were evaluated in situ after surface spraying. In WFH medium, the most active LAB isolates belonged to the Lactobacillus plantarum, reuteri and buchneri groups. The most active strains were added directly during “pains au lait” preparation and differents strains present delayed effect in particular a strain of Leuconostoc citreum which seems to delay the growth of Penicillium corylophilum after 10 days. In the second part, supernatants were analyzed to identified and quantified antifungal compounds by different treatments and different methods like HPLC, mass spectrometry. The results suggested that organic acids played the most important role in the antifungal activity and show that the main antifungal compounds corresponded to lactic, acetic and propionic acids, ethanol and hydrogen peroxide, as well as other compounds present at low levels. Based on these results, various combinations of the identified compounds were used to evaluate their effect on spore germination and fungal growth of P. corylophilum and E. repens. Some combinations presented the same activity than the bacterial culture supernatant thus confirming the involvement of the molecules in the antifungal activity. The results suggested that acetic acid was responsible of the entire antifungal activity against Penicillium corylophilum and played an important role in Eurotium repens inhibition. The selected bacteria provide a future prospect for use as bioprotective cultures on bakery products.