Comparative Genomic Analysis of Human Fungal Pathogens Causing Paracoccidioidomycosis

Paracoccidioides is a fungal pathogen and the cause of paracoccidioidomycosis, a health-threatening human systemic mycosis endemic to Latin America. Infection by Paracoccidioides, a dimorphic fungus in the order Onygenales, is coupled with a thermally regulated transition from a soil-dwelling filamentous form to a yeast-like pathogenic form. To better understand the genetic basis of growth and pathogenicity in Paracoccidioides, we sequenced the genomes of two strains of Paracoccidioides brasiliensis (Pb03 and Pb18) and one strain of Paracoccidioides lutzii (Pb01). These genomes range in size from 29.1 Mb to 32.9 Mb and encode 7,610 to 8,130 genes. To enable genetic studies, we mapped 94% of the P. brasiliensis Pb18 assembly onto five chromosomes. We characterized gene family content across Onygenales and related fungi, and within Paracoccidioides we found expansions of the fungal-specific kinase family FunK1. Additionally, the Onygenales have lost many genes involved in carbohydrate metabolism and fewer genes involved in protein metabolism, resulting in a higher ratio of proteases to carbohydrate active enzymes in the Onygenales than their relatives. To determine if gene content correlated with growth on different substrates, we screened the non-pathogenic onygenale Uncinocarpus reesii, which has orthologs for 91% of Paracoccidioides metabolic genes, for growth on 190 carbon sources. U. reesii showed growth on a limited range of carbohydrates, primarily basic plant sugars and cell wall components; this suggests that Onygenales, including dimorphic fungi, can degrade cellulosic plant material in the soil. In addition, U. reesii grew on gelatin and a wide range of dipeptides and amino acids, indicating a preference for proteinaceous growth substrates over carbohydrates, which may enable these fungi to also degrade animal biomass. These capabilities for degrading plant and animal substrates suggest a duality in lifestyle that could enable pathogenic species of Onygenales to transfer from soil to animal hosts.National Institute of Allergy and Infectious Diseases (U.S.)National Institutes of Health. Department of Health and Human Services (contract HHSN266200400001C)National Institutes of Health. Department of Health and Human Services(contract HHSN2722009000018C)Brazil. National Council for Scientific and Technological Developmen

Transmission of composite polymerization contraction force through a flowable composite and a resin-modified glass ionomer cement

The purpose of this study was to evaluate the individual contraction force during polymerization of a composite resin (Z-250), a flowable composite (Filtek Flow, FF) and a resin-modified glass ionomer cement (Vitrebond, VB), and the transmission of Z-250 composite resin polymerization contraction force through different thicknesses of FF and VB. The experiment setup consisted of two identical parallel steel plates connected to a universal testing machine. One was fixed to a transversal base and the other to the equipment's cross head. The evaluated materials were inserted into a 1-mm space between the steel plates or between the inferior steel plate and a previously polymerized layer of an intermediate material (either FF or VB) adhered to the upper steel plate. The composite resin was light-cured with a halogen lamp with light intensity of 500 mW/cm² for 60 s. A force/time graph was obtained for each sample for up to 120 s. Seven groups of 10 specimens each were evaluated: G1: Z-250; G2: FF; G3: VB; G4: Z-250 through a 0.5-mm layer of FF; G5: Z-250 through a 1-mm layer of FF; G6: Z-250 through a 0.5-mm of VB; G7: Z-250 through a 1-mm layer of VB. They were averaged and compared using one-way ANOVA and Tukey test at a = 0.05. The obtained contraction forces were: G1: 6.3N + 0.2N; G2: 9.8 + 0.2N; G3: 1.8 + 0.2N; G4: 6.8N + 0.2N; G5: 6.9N + 0.3N; G6: 4.0N + 0.4N and G7: 2.8N + 0.4N. The use of VB as an intermediate layer promoted a significant decrease in polymerization contraction force values of the restorative system, regardless of material thickness. The use of FF as an intermediate layer promoted an increase in polymerization contraction force values with both material thicknesses