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

A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

Meeting abstrac

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Search for a very light NMSSM Higgs boson produced in decays of the 125 GeV scalar boson and decaying into tau leptons in pp collisions at root 8=TeV

Peer reviewe

Reconstrução abdominal tardia sem tensão após laparostomia: uma nova técnica

OBJETIVO: A reconstrução abdominal tardia após laparostomia é sempre um procedimento desafiador para o cirurgião devido à necessidade de se corrigir um grande defeito na parede abdominal anterior, o que habitualmente demanda a lise de extensas aderências entre alças intestinais e o tecido cicatricial, sem que a hérnia incisional seja o resultado final. Neste trabalho, propomos uma técnica simples e inédita para esta reconstrução abdominal, sem tensão, utilizando tela de polipropileno sobre o tecido de granulação, sem necessidade de qualquer dissecção intra-peritoneal. MÉTODO: Descrição da técnica e estudo prospectivo de 17 pacientes submetidos à mesma entre 1998 e 2005. Foram analisados; a causa da laparostomia, o tempo entre a laparostomia e a reconstrução, o tempo operatório e a evolução pós-operatória imediata e tardia incluindo a incidência de hérnias incisionais. RESULTADOS: A idade média dos pacientes foi de 41 anos. As indicações da laparostomia foram; peritonite em oito pacientes, trauma abdominal em outros oito e pancreatite necrotizante em um. O tempo médio até a reconstrução abdominal foi de 14 meses. O tempo médio do procedimento cirúrgico foi de 130 minutos. O período médio de internação hospitalar foi de 2,6 dias para os 15 pacientes sem complicações pós-operatórias. Não houveram óbitos ou ocorrência de síndrome de compartimento abdominal relacionados à técnica. A média do período de acompanhamento pós-operatório é de 24 meses e até o momento não há ocorrência de hérnia incisional em todo o grupo. CONCLUSÕES: A técnica aqui proposta é de fácil execução e reprodutibilidade, torna desnecessária a manipulação da cavidade abdominal com conseqüente diminuição do risco de lesão de vísceras abdominais e proporciona o fechamento definitivo da laparostomia sem tensão. Esta técnica não acarretou síndrome de compartimento abdominal e nenhum paciente desenvolveu hérnia incisional até o momento

Influence of luting agents on time required for cast post removal by ultrasound: an in vitro study

OBJECTIVE: This in vitro study evaluated the influence of luting agents on ultrasonic vibration time for intraradicular cast post removal. MATERIAL AND METHODS: After endodontic treatment, 30 roots of extracted human canines were embedded in resin cylinders. The post-holes were prepared at 10 mm depth and their impressions were taken using autopolymerizing acrylic resin. After casting procedures using a nickel-chromium alloy, the posts were randomly distributed into 3 groups (n=10) according to the luting material: G1- zinc phosphate (SS White) (control group), G2 - glass ionomer cement (Vidrion C; SS White), and G3- resin cement (C&B; Bisco). In G3, the adhesive procedure was performed before post cementation. After 24 h, the cement line was removed at the post/tooth interface using a fine diamond bur, and the ST-09 tip of an Enac ultrasound unit was applied at maximum power on all surfaces surrounding the posts. The application time was recorded with a chronometer until the post was completely dislodged and data were analyzed by ANOVA and Tukey's test (p<0.05). RESULTS: The roots were removed from the acrylic resin and inspected to detect cracks and/or fractures. The means for G1, G2, and G3 were 168.5, 59.5, and 285 s, respectively, with statistically significant differences among them. Two G3 posts resisted removal, one of which developed a vertical fracture line. CONCLUSIONS: Therefore, the cement type had a direct influence on the time required for ultrasonic post removal. Compared to the zinc phosphate and glass ionomer cements, the resin cement required a longer ultrasonic vibration time