Secondary mineral formation associated with respiration of nontronite, NAu-1 by iron reducing bacteria

Experimental batch and miscible-flow cultures were studied in order to determine the mechanistic pathways of microbial Fe(III) respiration in ferruginous smectite clay, NAu-1. The primary purpose was to resolve if alteration of smectite and release of Fe precedes microbial respiration. Alteration of NAu-1, represented by the morphological and mineralogical changes, occurred regardless of the extent of microbial Fe(III) reduction in all of our experimental systems, including those that contained heat-killed bacteria and those in which O(2), rather than Fe(III), was the primary terminal electron acceptor. The solid alteration products observed under transmission electron microscopy included poorly crystalline smectite with diffuse electron diffraction signals, discrete grains of Fe-free amorphous aluminosilicate with increased Al/Si ratio, Fe-rich grains, and amorphous Si globules in the immediate vicinity of bacterial cells and extracellular polymeric substances. In reducing systems, Fe was also found as siderite. The small amount of Fe partitioned to the aqueous phase was primarily in the form of dissolved Fe(III) species even in the systems in which Fe(III) was the primary terminal electron acceptor for microbial respiration. From these observations, we conclude that microbial respiration of Fe(III) in our laboratory systems proceeded through the following: (1) alteration of NAu-1 and concurrent release of Fe(III) from the octahedral sheets of NAu-1; and (2) subsequent microbial respiration of Fe(III)

The use of contextualised standardised client simulation to develop clinical reasoning in final year veterinary students

Clinical reasoning is an important skill for veterinary students to develop before graduation. Simulation has been studied in medical education as a method for developing clinical reasoning in students, but evidence supporting it is limited. This study involved the creation of a contextualized, standardized client simulation session that aimed to improve the clinical reasoning ability and confidence of final-year veterinary students. Sixty-eight participants completed three simulated primary-care consultations, with the client played by an actor and the pet by a healthy animal. Survey data showed that all participants felt that the session improved their clinical decision-making ability. Quantitative clinical reasoning self-assessment, performed using a validated rubric, triangulated this finding, showing an improvement in students’ perception of several components of their clinical reasoning skill level from before the simulation to after it. Blinded researcher analysis of the consultation video recordings found that students showed a significant increase in ability on the history-taking and making-sense-of-data (including formation of a differential diagnosis) components of the assessment rubric. Thirty students took part in focus groups investigating their experience with the simulation. Two themes arose from thematic analysis of these data: variety of reasoning methods and “It’s a different way of thinking.” The latter highlights differences between the decision making students practice during their time in education and the decision making they will use once they are in practice. Our findings suggest that simulation can be used to develop clinical reasoning in veterinary students, and they demonstrate the need for further research in this area

Exchange of functional domains between a bacterial conjugative relaxase and the integrase of the human adeno-associated virus

Endonucleases of the HUH family are specialized in processing single-stranded DNA in a variety of evolutionarily highly conserved biological processes related to mobile genetic elements. They share a structurally defined catalytic domain for site-specific nicking and strand-transfer reactions, which is often linked to the activities of additional functional domains, contributing to their overall versatility. To assess if these HUH domains could be interchanged, we created a chimeric protein from two distantly related HUH endonucleases, containing the N-terminal HUH domain of the bacterial conjugative relaxase TrwC and the C-terminal DNA helicase domain of the human adeno-associated virus (AAV) replicase and site-specific integrase. The purified chimeric protein retained oligomerization properties and DNA helicase activities similar to Rep68, while its DNA binding specificity and cleaving-joining activity at oriT was similar to TrwC. Interestingly, the chimeric protein could catalyse site-specific integration in bacteria with an efficiency comparable to that of TrwC, while the HUH domain of TrwC alone was unable to catalyze this reaction, implying that the Rep68 C-terminal helicase domain is complementing the TrwC HUH domain to achieve site-specific integration into TrwC targets in bacteria. Our results illustrate how HUH domains could have acquired through evolution other domains in order to attain new roles, contributing to the functional flexibility observed in this protein superfamily.This work was supported by the Medical Research Council (MRC) grant MR/N022890/1 to EH and grant 1001764 to RML; National Institutes of Health (NIH) grant RO1-GM09285 to CRE; Spanish Ministry of Economy and competitiveness (MINECO) grant BIO2013-46414-P to ML and AFM is supported by a Doc.Mobility fellowship from the Swiss National Science Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

High-Anxious Individuals Show Increased Chronic Stress Burden, Decreased Protective Immunity, and Increased Cancer Progression in a Mouse Model of Squamous Cell Carcinoma

In spite of widespread anecdotal and scientific evidence much remains to be understood about the long-suspected connection between psychological factors and susceptibility to cancer. The skin is the most common site of cancer, accounting for nearly half of all cancers in the US, with approximately 2–3 million cases of non-melanoma cancers occurring each year worldwide. We hypothesized that a high-anxious, stress-prone behavioral phenotype would result in a higher chronic stress burden, lower protective-immunity, and increased progression of the immuno-responsive skin cancer, squamous cell carcinoma. SKH1 mice were phenotyped as high- or low-anxious at baseline, and subsequently exposed to ultraviolet-B light (1 minimal erythemal dose (MED), 3 times/week, 10-weeks). The significant strengths of this cancer model are that it uses a normal, immunocompetent, outbred strain, without surgery/injection of exogenous tumor cells/cell lines, and produces lesions that resemble human tumors. Tumors were counted weekly (primary outcome), and tissues collected during early and late phases of tumor development. Chemokine/cytokine gene-expression was quantified by PCR, tumor-infiltrating helper (Th), cytolytic (CTL), and regulatory (Treg) T cells by immunohistochemistry, lymph node T and B cells by flow cytometry, adrenal and plasma corticosterone and tissue vascular-endothelial-growth-factor (VEGF) by ELISA. High-anxious mice showed a higher tumor burden during all phases of tumor development. They also showed: higher corticosterone levels (indicating greater chronic stress burden), increased CCL22 expression and Treg infiltration (increased tumor-recruited immuno-suppression), lower CTACK/CCL27, IL-12, and IFN-γ gene-expression and lower numbers of tumor infiltrating Th and CTLs (suppressed protective immunity), and higher VEGF concentrations (increased tumor angiogenesis/invasion/metastasis). These results suggest that the deleterious effects of high trait anxiety could be: exacerbated by life-stressors, accentuated by the stress of cancer diagnosis/treatment, and mediate increased tumor progression and/or metastasis. Therefore, it may be beneficial to investigate the use of chemotherapy-compatible anxiolytic treatments immediately following cancer diagnosis, and during cancer treatment/survivorship

A user's guide to the Encyclopedia of DNA elements (ENCODE)

The mission of the Encyclopedia of DNA Elements (ENCODE) Project is to enable the scientific and medical communities to interpret the human genome sequence and apply it to understand human biology and improve health. The ENCODE Consortium is integrating multiple technologies and approaches in a collective effort to discover and define the functional elements encoded in the human genome, including genes, transcripts, and transcriptional regulatory regions, together with their attendant chromatin states and DNA methylation patterns. In the process, standards to ensure high-quality data have been implemented, and novel algorithms have been developed to facilitate analysis. Data and derived results are made available through a freely accessible database. Here we provide an overview of the project and the resources it is generating and illustrate the application of ENCODE data to interpret the human genome

209Bi NMR and NQR investigation of the small-gap semiconductor Ce3Bi4Pt3.

We report measurements of the temperature dependence of the Bi209 nuclear quadrupole resonance frequency νQ, the Knight shift K, and the spin-lattice relaxation rate 1/T1 in the small-gap semiconductor Ce3Bi4Pt3 between 1.8 and 300 K. Corresponding measurements also are reported for the nonmagnetic metallic isomorph La3Bi4Pt3. The νQ data in the Ce compound show a characteristic departure from metallic-to-insulating behavior when the sample is cooled below TM=80 K, the temperature of the susceptibility maximum, attributable to a loss of low-frequency vibrational modes in the insulating state. The Knight shift has both isotropic and axial components; this anisotropy originates from the presence of Ce via a transferred hyperfine coupling between Ce 4f and conduction electrons. An s-f exchange constant 0.4 eV is found, consistent with hybridization in other rare-earth intermetallic compounds. A change in the scaling between the susceptibility and both the isotropic and axial Knight shifts at temperature TM provides evidence that hybridization between the Ce 4f orbitals and the conduction electrons is responsible for the gap structure. The temperature dependence of the 1/T1 data is consistent with a model electronic density of states possessing a temperature-independent gap δ of 180 K and a bandwidth of the order of 1600 K. The temperature dependence of 1/T1 can also be fit well with a temperature-dependent gap with δ(0) also 180 K. © 1994 The American Physical Society

209Bi NMR in the heavy-electron system YbBiPt

Bismuth NMR Knight shift and spin lattice relaxation rate 1/T1 data are reported between 35 and 325 K in the low-carrier heavy fermion system YbBiPt. The Knight shift is strongly temperature dependent and negative. Its temperature dependence tracks the bulk susceptibility with a hyperfine coupling constant Ahf = -7.88 kOe/μB. At low temperatures 1/T1 exhibits a dramatic increase, such that the average 4f spin correlation time τf shows a crossover behavior at about 75 K. The rate 1 τf is proportional to temperature above 75 K, consistent with non-interacting 4f local moments which are relaxed via Korringa-type scattering with the conduction electrons. We discuss the behavior below 75 K in terms of crystal-field effects or a strongly temperature dependent contribution from non-zero q regime of the dynamical susceptibility. © 1995

209Bi NMR in the heavy-electron system YbBiPt

Bismuth NMR Knight shift and spin lattice relaxation rate 1/T1 data are reported between 35 and 325 K in the low-carrier heavy fermion system YbBiPt. The Knight shift is strongly temperature dependent and negative. Its temperature dependence tracks the bulk susceptibility with a hyperfine coupling constant Ahf = -7.88 kOe/μB. At low temperatures 1/T1 exhibits a dramatic increase, such that the average 4f spin correlation time τf shows a crossover behavior at about 75 K. The rate 1 τf is proportional to temperature above 75 K, consistent with non-interacting 4f local moments which are relaxed via Korringa-type scattering with the conduction electrons. We discuss the behavior below 75 K in terms of crystal-field effects or a strongly temperature dependent contribution from non-zero q regime of the dynamical susceptibility. © 1995