Abstract：
Paracoccidioidomycosis (PCM) is a systemic mycosis caused by Paracoccidioides spp. The interaction mediated by the presence of adhesins on the fungal surface and receptors in the extracellular matrix of the host, as well as the biofilm formation, is essential in its pathogenesis. Adhesins such as gp43, enolase, GAPDH (glyceraldehyde-3-phosphate dehydrogenase), and 14-3-3 have been demonstrated in the Paracoccidioides brasiliensis (Pb18) strain and recognized as necessary in the fungus-host interaction. The Pb 18 strain silenced to 14-3-3 showed changes in morphology, virulence, and adhesion capacity. The study aimed to evaluate the role of adhesin 14-3-3 in P. brasiliensis biofilm formation and the differential expression of genes related to adhesins, comparing planktonic and biofilm forms. The presence of biofilm was also verified in sutures in vitro and in vivo. The silenced strain (Pb14-3-3 aRNA) was compared with the wild type Pb18, determining the differential metabolic activity between the strains by the XTT reduction assay; the biomass by violet crystal and the polysaccharides by safranin, even as morphological differences by microscopic techniques. Differential gene expression for adhesins was also analyzed, comparing the relative expression of these in planktonic and biofilm forms at different times. The results suggested that the silencing of 14-3-3 protein altered the ability to form biofilm and its metabolism. The quantity of biomass was similar in both strains; however, the formation of exopolymeric substances and polysaccharide material was lower in the silenced strain. Our results showed increased expression of enolase, GAPDH, and 14-3-3 genes in the first periods of biofilm formation in the Pb18 strain. In contrast, the silenced strain showed a lower expression of these genes, indicating that gene silencing can influence the expression of other genes and be involved in the biofilm formation of P. brasiliensis. In vitro and in vivo assays using sutures confirmed this yeast\'s ability to form biofilm and may be implicated in the pathogenesis of paracoccidioidomycosis.
摘要：
副角菌病（PCM）是由副角菌病引起的全身性真菌病。由真菌表面粘附素和宿主细胞外基质中受体的存在介导的相互作用，以及生物膜的形成，在其发病机理中至关重要。粘附素，如gp43，烯醇化酶，GAPDH（甘油醛-3-磷酸脱氢酶），和14-3-3-3已在巴西副球菌（Pb18）菌株中得到证明，并被认为是真菌与宿主相互作用所必需的。沉默为14-3-3的Pb18菌株显示出形态变化，毒力,和粘附能力。本研究旨在评估粘附素14-3-3在巴西巴氏杆菌生物膜形成中的作用以及与粘附素相关基因的差异表达。比较浮游和生物膜形式。生物膜的存在也在体外和体内的缝合线中得到证实。将沉默的菌株（Pb14-3-3aRNA）与野生型Pb18进行比较，通过XTT还原试验确定菌株之间的差异代谢活性；紫水晶的生物量和番红的多糖，甚至作为微观技术的形态差异。还分析了粘附素的差异基因表达，比较这些在不同时间浮游和生物膜形式中的相对表达。结果表明，14-3-3蛋白的沉默改变了生物膜的形成能力及其代谢。两种菌株的生物量相似；然而，在沉默的菌株中，外聚合物和多糖物质的形成较低。我们的结果显示烯醇化酶的表达增加，GAPDH,在Pb18菌株的生物膜形成的第一阶段和14-3-3基因。相比之下,沉默的菌株显示这些基因的表达较低，表明基因沉默可以影响其他基因的表达，并参与巴西假单胞菌的生物膜形成。使用缝合线的体外和体内试验证实了这种酵母形成生物膜的能力，并且可能与副球菌病的发病机理有关。
