923 resultados para 28S rDNA


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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Candida yeasts are common in the oral cavity and can cause candidosis in the presence of predisposing factors, especially diabetes. The manifestation of the disease is related to this set of local factors such as the presence of dental prostheses, salivary pH, salivary flow and tobacco and the ability to form biofilms. Biofilms are specific and organized communities of cells under the control of signaling molecules rather than random accumulations of cells resulting from cell division and frequently are drugs resistance. Aim: The objectives of this study were to determine the genetic patterns of these C. albicans isolates and to evaluate the in vitro activity amphotericin B and caspofungin against C. albicans biofilms. Methods: Microbial samples were collected from subgingival sites and seeded in CHROMagar for subsequent identification of C. albicans by PCR. Genotypes were defined based on the identification of the transposable introns in the 25S rDNA by PCR. Results: In this study, 6 strains were identified as C. albicans and of these, 3 strains were genotype A and 3 were genotype B. The results showed that both amphotericin B and caspofungin exhibited strong antifungal activities against C. albicans biofilm formation and inhibiting the biofilm formation ranging from 70.8 – 95.3% and 77.7 - 88.7%, respectively. The antifungals studied had low inhibitory effect on preformed biofims, ranging from 39.5 - 50.8% for amphotericin B and from 23.1 - 36.9% for caspofungin at the same concentration. The activity of the two drugs was most effective in inhibit biofilm formation.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Background: The intestinal microbiome (IM) has extensively been studied in the search for a link of bacteria with the cause of Crohn`s disease (CD). The association might result from the action of a specific pathogen and/or an eventual imbalance in bacterial species composition of the gut. The innumerous virulence associated markers and strategies described for adherent and invasive Escherichia coli (AIEC) have made them putative candidate pathogens for CD. IM of CD patients shows dysbiosis, manifested by the proliferation of bacterial groups such as Enterobacteriaceae and reduction of others such as Lactobacillus and Bifidobacterium. The augmented bacterial population comprising of commensal and/or pathogenic organisms super stimulates the immune system, triggering the inflammatory reactions responsible for the clinical manifestations of the disease. Considering the role played by IM in CD and the multiple variables influencing its species composition, resulting in differences among populations, the objective of this study was to determine the bacterial biodiversity in the mucosa associated microbiome of CD patients from a population not previously subject to this analysis, living in the middle west region of Sao Paulo state. Methods: A total of 4 CD patients and 5 controls subjects attending the Botucatu Medical School of the Sao Paulo State University (UNESP) for routine colonoscopy and who signed an informed consent were included in the study. A number of 2 biopsies, one from the ileum and other from any part of the terminal colon, were taken from each subject and immediately frozen at -70[degrees]C until DNA purification. The bacterial biodiversity was assessed by next generation (ion torrent) sequencing of PCR amplicons of the ribosomal DNA 16S V6 region (16S V6 rDNA). The bacterial identification was performed at the genus level, by alignment of the generated DNA sequences with those available at the ribosomal database project (RDP) website. Results: The overall DNA sequence output was based on an average number of 526,427 reads per run, matching 50 bacterial genus 16SrDNA sequences available at the RDB website, and 22 non matching sequences. Over 95% of the sequences corresponded to taxa belonging to the major phyla: Firmicutes, Bacterioidetes, Proteobacteria and Actinobacteria. Irrespective of the intestinal site analyzed, no case-control differences could be observed in the prevalence of Actinobacteria and Firmicutes. The prevalence of Proteobacteria was higher (40%) in the biopsies of control subjects as compared to that of DC patients (16%). For Bacterioidetes, the higher prevalence was observed among DC patients (33% as opposed to 14,5% in controls). The significance for all comparisons considered a p value < 0,05 in a Chi2 test. No mucosal site specific differences could be observed in IM comparisons of CD and control subjects. Conclusions: The rise in the number of Bacterioidetes observed here among CD patients seems to be in agreement with most of studies published thus far. Yet, the reduction in the number of Proteobacteria along with an apparently unaltered population of Actinobacteria and Firmicutes, which include the so called "beneficial" organisms Bifidobacterium and Lactobacillus were rather surprising. These data suggest that the analyses on the role of IM in CD should consider the multiple variables that may influence its species composition.

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As formigas do gênero Cephalotes, rapidamente identificadas por suas operárias polimórficas de cutícula resistente e cabeça achatada possuem seu sucesso ecológico creditado à sua dieta predominantemente generalista e nidificação em cavidades pré-existentes de troncos de árvores. Possuem hábitos exclusivamente arborícolas e ocorrem nos trópicos e subtrópicos do Novo Mundo possuindo ampla distribuição geográfica. O grupo apresenta uma interessante associação com microorganismos. No presente trabalho foi feita a caracterização molecular e estudo de relações filogenéticas do gênero através da amplificação e sequenciamento de fragmento do gene 28S do DNA Nuclear de três populações localizadas em Rio Claro-SP e São José do Rio Preto-SP de duas espécies de Cephalotes: C. pusillus e C. clypeatus. Também foi feito o levantamento da ocorrência e frequência do endossimbionte Wolbachia em sete populações de Cephalotes localizadas em São José do Rio Preto-SP, Guaraci-SP, São Carlos-SP, Araraquara-SP, Delfinópolis-MG e Rio Claro-SP; abrangendo três espécies: C. pusillus, C. clypeatus e C. atratus. Esse levantamento foi realizado com a utilização de ferramentas moleculares para a análise do gene codificador da proteína de superfície de membrana do endossimbionte, o wsp. Para analises de filogenia e também do endossimbionte, foi realizada a extração do DNA total de operárias, a amplificação do gene através da técnica de PCR utilizando os primers já estabelecidos e em seguida, as amostras foram sequenciadas pelo método de Sanger. Os resultados obtidos mostraram relações monofiléticas dentro da subfamília Myrmicinae, a qual pertence o gênero Cephalotes. As análises do gene 28S trouxeram resultados otimistas no estudo da caracterização molecular do grupo. Os resultados da analise do endossimbionte corroboraram com estudos anteriores com outras espécies do gênero Cephalotes, onde ocorreu alta...

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Black fungi are able to adapt to extreme environmental conditions, such as: high temperatures, the presence of toxic chemical substances and lack of nutrients. Besides, they are also potential pathogens to humans. The natural environment of many black fungi is still unknown and some studies are being conducted to evaluate the biodiversity of this group and their different habitats. This study aimed to isolate black fungi in domestic environments and facilities, such as toothbrushes, fridge sealing rubbers, bathroom strainers and divisions, windows, wall tiles and bath sponge. For the collection, material surfaces were scratched with a scalpel and the resulting fragments were sewed in Mycosel agar (DifcoTM), supplemented with actidione to inhibit the growth of highly-sporulating fungi. Plates were incubated at 25ºC for three weeks. The 46 isolated fungi were maintained on MA2% slants at 8ºC and cryopreserved at -80ºC. Fungal identification was performed through the analysis of macro and microscopic features and ITS rDNA sequencing. The following black fungi taxa were found: Ascomycota sp., Cladosporium spp., Dothideomycete sp., Exophiala alcalophila, Ochroconis mirabilis and Rhinocladiella atrovirens. Non-melanized fungi were also found, such as Geosmithia sp., Penicillium sp. and Rhodotorula mucilaginosa. The temperature tests showed that isolated black fungi were not able to grow at 37°C, however, this temperature proved to be fungistatic to 43% of them. According to literature, all black fungi isolated in this study are opportunistic pathogens and additional studies are necessary to evaluate the risk that these micro-organisms offer to health, once they were isolated from domestic environments

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