Analyzing the Role of a Protein Downregulated After Induction of Filamentous Growth In Candida albicans

Candida albicans is a commensal fungus, normally living with its human host, however, it has the ability to cause invasive infection. Candida albicans is the fourth most frequent nosocomial infection affecting a vulnerable immunocompromised population. Candida albicans exhibits different morphologies including yeast, pseudohyphae, and hyphae. The varying morphological potential of this organism is a virulence trait. Because of this, research has focused on what drives activation of hyphal formation as well as what impedes it. During a filamentation assay, a novel observation pertaining to a subgroup of proteins being downregulated early after germination, was made. In this study, we constructed an over-expression strain of one of these proteins and have used known hypha inducing media to determine if it will have an impact on filamentation. With the conditions tested thus far, no significant impacts on morphology have been observed

A Symplectic Integrator for Hill's Equations

Hill's equations are an approximation that is useful in a number of areas of astrophysics including planetary rings and planetesimal disks. We derive a symplectic method for integrating Hill's equations based on a generalized leapfrog. This method is implemented in the parallel N-body code, PKDGRAV and tested on some simple orbits. The method demonstrates a lack of secular changes in orbital elements, making it a very useful technique for integrating Hill's equations over many dynamical times. Furthermore, the method allows for efficient collision searching using linear extrapolation of particle positions.Comment: 15 pages, 2 figures; minor revisions; accepted for publication in the Astronomical Journa

Outcomes following oesophagectomy in patients with oesophageal cancer: a secondary analysis of the ICNARC Case Mix Programme Database

Introduction: This report describes the case mix and outcomes of patients with oesophageal cancer admitted to adult critical care units following elective oesophageal surgery in England, Wales and Northern Ireland. Methods: Admissions to critical care following elective oesophageal surgery for malignancy were identified using data from the Intensive Care National Audit and Research Centre (ICNARC) Case Mix Programme Database. Information on admissions between December 1995 and September 2007 were extracted and the association between in-hospital mortality and patient characteristics on admission to critical care was assessed using multiple logistic regression analysis. The performance of three prognostic models (Simplified Acute Physiology Score (SAPS) II, Acute Physiology and Chronic Health Evaluation (APACHE) II and the ICNARC physiology score) was also evaluated. Results: Between 1995 and 2007, there were 7227 admissions to 181 critical care units following oesophageal surgery for malignancy. Overall mortality in critical care was 4.4% and in-hospital mortality was 11%, although both declined steadily over time. Eight hundred and seventy-three (12.2%) patients were readmitted to critical care, most commonly for respiratory complications (49%) and surgical complications (25%). Readmitted patients had a critical care unit mortality of 24.7% and in-hospital mortality of 33.9%. Overall in-hospital mortality was associated with patient age, and various physiological measurements on admission to critical care (partial pressure of arterial oxygen (PaO2):fraction of inspired oxygen (FiO2) ratio, lowest arterial pH, mechanical ventilation, serum albumin, urea and creatinine). The three prognostic models evaluated performed poorly in measures of discrimination, calibration and goodness of fit. Conclusions: Surgery for oesophageal malignancy continues to be associated with significant morbidity and mortality. Age and organ dysfunction in the early postoperative period are associated with an increased risk of death. Postoperative serum albumin is confirmed as an additional prognostic factor. More work is required to determine how this knowledge may improve clinical management

From Isotopes to TK Interviews: Towards Interdisciplinary Research in Fort Resolution and the Slave River Delta, Northwest Territories

Evolving research in Fort Resolution and the Slave River Delta, Northwest Territories, aims to improve understanding of how the natural ecosystem functions and responds to various environmental stressors, as well as to enhance the stewardship of natural resources and the capacity of local residents to respond to change. We seek to integrate approaches that span the natural and social sciences and traditional knowledge understandings of change, employing a research design developed in response to the concerns of a northern community. In doing so, we have strived for a research process that is collaborative, interdisciplinary, policy-oriented, and reflective of northern priorities. These elements characterize the new northern research paradigm increasingly promoted by various federal funding agencies, northern partners, and communities. They represent a holistic perspective in the pursuit of solutions to address complex environmental and socioeconomic concerns about impacts of climate change and resource development on northern societies. However, efforts to fulfill the objectives of this research paradigm are associated with a host of on-the-ground challenges. These challenges include (but are not restricted to) developing effective community partnerships and collaboration and documenting change through interdisciplinary approaches. Here we provide an overview of the components that comprise our interdisciplinary research program and offer an accounting of our formative experiences in confronting these challenges

Ex vivo renal perfusion and autotransplantation in treatment of calculous disease or abdominal aortic aneurysm.

Two more indications are described for temporary ex vivo perfusion of kidneys with revascularization of these organs as autografts to orthotopic or heterotopic locations. One of the patients had staghorn calculi which were removed from a solitary kidney. The other patient had both kidneys autografted in the course of a surgical procedure on an extensive abdominal aortic aneurysm

Absence of \u3ci\u3esodA\u3c/i\u3e Increases the Levels of Oxidation of Key Metabolic Determinants of \u3ci\u3eBorrelia burgdorferi\u3c/i\u3e

Borrelia burgdorferi, the causative agent of Lyme disease, alters its gene expression in response to environmental signals unique to its tick vector or vertebrate hosts. B. burgdorferi carries one superoxide dismutase gene (sodA) capable of controlling intracellular superoxide levels. Previously, sodA was shown to be essential for infection of B. burgdorferi in the C3H/HeN model of Lyme disease. We employed two-dimensional electrophoresis (2-DE) and immunoblot analysis with antibodies specific to carbonylated proteins to identify targets that were differentially oxidized in the soluble fractions of the sodA mutant compared to its isogenic parental control strain following treatment with an endogenous superoxide generator, methyl viologen (MV, paraquat). HPLC-ESI-MS/MS analysis of oxidized proteins revealed that several proteins of the glycolytic pathway (BB0057, BB0020, BB0348) exhibited increased carbonylation in the sodA mutant treated with MV. Levels of ATP and NAD/NADH were reduced in the sodA mutant compared with the parental strain following treatment with MV and could be attributed to increased levels of oxidation of proteins of the glycolytic pathway. In addition, a chaperone, HtpG (BB0560), and outer surface protein A (OspA, BBA15) were also observed to be oxidized in the sodA mutant. Immunoblot analysis revealed reduced levels of Outer surface protein C (OspC), Decorin binding protein A (DbpA), fibronectin binding protein (BBK32), RpoS and BosR in the sodA mutant compared to the control strains. Viable sodA mutant spirochetes could not be recovered from both gp91/phox−⁄− and iNOS deficient mice while borrelial DNA was detected in multiple tissues samples from infected mice at significantly lower levels compared to the parental strain. Taken together, these observations indicate that the increased oxidation of select borrelial determinants and reduced levels of critical pathogenesis-associated lipoproteins contribute to the in vivo deficit of the sodA mutant in the mouse model of Lyme disease. This study, utilizing the sodA mutant, has provided insights into adaptive capabilities critical for survival of B. burgdorferi in its hosts

ENZYME ENGINEERING AT ALMAC: Case studies of enzyme discovery and engineering

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Presence of extracellular DNA in candida albicans biofilm matrix and its role in biofilm structure and antifungal susceptibility

Biofilms are structurally complex microconsortia of surface adhering cells embedded within an extracellular matrix (ECM) composed of substances produced and secreted by cells or derived from cell lysis. One of the recently discovered bacterial biofilms ECM components is the extracellular DNA (eDNA). Although the investigation on eDNA in fungal biofilms is scarce, preliminary studies suggest that eDNA may play a role in biofilms formed by the opportunistic fungal pathogen Candida albicans. Thus, the present study aimed at determining the eDNA content of C. albicans SC5314 biofilm ECM and the effect of DNase I treatment on biofilm formation and biofilm cells susceptibility to antifungals, as indicators of the role of eDNA in C. albicans biofilms. Results from our experiments showed that the ECM of C. albicans biofilms formed under conditions of flow for 48 h contained 3045.4 ± 227.3 ng eDNA/mg of protein. Additionally, using a microtiter plate model, we observed that different DNase treatments (0.02 - 2 mg/ml) did not affect further biofilm development by C. albicans adherent cells. However, DNase (> 0.03 mg/ml) promoted a general biomass reduction on C. albicans preformed biofilms. Finally, DNase (0.13 mg/ml) did not change C. albicans biofilm cells susceptibility to fluconazole, but increased their susceptibility to amphotericin B and caspofungin, as indicated by the lower SMIC compared to biofilms grown without DNase. This work presents evidence for the role of eDNA in C. albicans biofilm integrity and antifungal resistance consistent with eDNA being a key element of the ECM

Development and application of novel engineered transaminase panels assisted by in- silico rational design for the production of chiral amines

There is a high demand for the synthesis of chiral amines as building blocks for a large number of industrially valuable compounds. Transaminases (TAm) offer an enzymatic route for the synthesis of chiral amines that avoids complex chemical synthesis [1]. However, their catalytic efficiency towards bulky ketone substrates is greatly limited by steric hinderance [2]. This poster highlights a rational design strategy of combining in silico and in vitro methods to engineer the transaminase enzyme with a minimal number of mutations, achieving high catalytic activity and high enantioselectivity. The wildtype TAm showed no detectable activity towards the ketone 2-acetylbiphenyl but upon introduction of two mutations detectable enzyme activity was observed. The reaction rate was improved a further 1716-fold with the rationally designed variant, that contained a further 5 mutations, producing the corresponding enantiomeric pure (S)-amine (enantiomeric excess (ee) value of \u3e99%)[3]. In addition, screening of in silico designed (R)-TAm mutant panels in resolution mode offered an attractive and efficient route for the preparation of problematic (S)-amines. A mutant was identified from the panels that gave complete resolution of the racemic amine (high substrate loading) to leave the desired enantiomer at a low enzyme loading fit for process development towards an economically viable scale up process. [1] R. C. Simon, et al, ACS Catal. 2014, 4(1) [2] F. Steffen-Munsberg, et al, ChemCatChem 2013, 5, (1) [3]D.F.A.R.Dourado et al, ACS Catal. 2016, 6 (11