Hsc66 substrate specificity is directed toward a discrete region of the iron-sulfur cluster template protein IscU

Hsc66 and Hsc20 comprise a specialized chaperone system important for the assembly of iron-sulfur clusters in Escherchia coli. Only a single substrate, the Fe/S template protein IscU, has been identified for the Hsc66/Hsc20 system, but the mechanism by which Hsc66 selectively binds IscU is unknown. We have investigated Hsc66 substrate specificity using phage display and a peptide array of IscU. Screening of a heptameric peptide phage display library revealed that Hsc66 prefers peptides with a centrally located Pro-Pro motif. Using a cellulose-bound peptide array of IscU we determined that Hsc66 interacts specifically with a region (residues 99-103, LPPVK) that is invariant among all IscU family members. A synthetic peptide (ELPPVKIHC) corresponding to IscU residues 98-106 behaves in a similar manner to native IscU, stimulating the ATPase activity of Hsc66 with similar affinity as IscU, preventing Hsc66 suppression of bovine rhodanese aggregation, and interacting with the peptide-binding domain of Hsc66. Unlike native IscU, however, the synthetic peptide is not bound by Hsc20 and does not synergistically stimulate Hsc66 ATPase activity with Hsc20. Our results indicate that Hsc66 and Hsc20 recognize distinct regions of IscU and further suggest that Hsc66 will not bind LPPVK motifs with high affinity in vivo unless they are in the context of native IscU and can be directed to Hsc66 by Hsc20

Microstructural evolution in infiltration-growth processed MgB₂ bulk superconductors

The study reports phase and microstructural evolution in MgB2 bulk superconductors fabricated by an infiltration and growth (IG) process. Three distinct stages, (1) intermediate boride formation, (2) bulk liquid Mg infiltration, and (3) MgB2 layer formation, were identified in IG process after detailed examination of series of samples prepared with varied heating conditions. The intermediate phase Mg2B25, isomorphous to β-boron, was detected prior to MgB2 phase formation in stage (1). Due to volume expansion involved in stage 1, cracks formed in the β-boron particles and propagated radially inwards during stage 3. The growing MgB2 particles sintered simultaneously with formation of grain boundaries during the process, as evidenced by the measured hardness and critical current density in these samples. From our observations, we estimate the total time needed for complete transformation to MgB2.Authors acknowledge financial support from KACST-Cambridge Joint Centre of Excellence in Advanced Materials and Manufacturing (CAMM) based at the University of Cambridge, UK. Partial financial support from Engineering and Physical Sciences Research Council, UK (Grant: EP/K031422/1) is gratefully acknowledged

Microstructural evolution in infiltration-growth processed MgB2 bulk superconductors

© 2017 The Authors. The study reports phase and microstructural evolution in MgB2 bulk superconductors fabricated by an Infiltration and Growth (IG) process. Three distinct stages: (1) intermediate boride formation (2) bulk liquid Mg infiltration and (3) MgB2 layer formation, were identified in IG process after detailed examination of series of samples prepared with varied heating conditions. The intermediate phase Mg2B25, isomorphous to β-Boron, was detected prior to MgB2 phase formation in stage (1). Due to volume expansion involved in stage 1, cracks formed in the β-Boron particles and propagated radially inwards during stage 3. The growing MgB2 particles sintered simultaneously with formation of grain boundaries during the process, as evidenced by the measured hardness and critical current density in these samples. From our observations we estimate the total time needed for complete transformation to MgB2.Authors acknowledge financial support from KACST-Cambridge Joint Centre of Excellence in Advanced Materials and Manufacturing (CAMM) based at the University of Cambridge, UK. Partial financial support from Engineering and Physical Sciences Research Council, UK (Grant: EP/K031422/1) is gratefully acknowledged.KACST‐Cambridge Joint Centre of Excellence in Advanced Materials and Manufacturing; Engineering and Physical Sciences Research Council. Grant Number: EP/K031422/

Imaging characteristics and treatment of a penetrating brain injury caused by an oropharyngeal foreign body in a dog

A 4-year-old Border collie was presented with one episode of collapse, altered mentation, and a suspected pharyngeal stick injury. Magnetic resonance imaging (MRI) and computed tomography showed a linear foreign body penetrating the right oropharynx, through the foramen ovale and the brain parenchyma. The foreign body was surgically removed and medical treatment initiated. Complete resolution of clinical signs was noted at recheck 8 weeks later. Repeat MRI showed chronic secondary changes in the brain parenchyma. To the authors' knowledge, this is the first report of the advanced imaging findings and successful treatment of a penetrating oropharyngeal intracranial foreign body in a dog

Three hydrophobic amino acids in Escherichia coli HscB make the greatest contribution to the stability of the HscB-IscU complex

<p>Abstract</p> <p>Background</p> <p>General iron-sulfur cluster biosynthesis proceeds through assembly of a transient cluster on IscU followed by its transfer to a recipient apo-protein. The efficiency of the second step is increased by the presence of HscA and HscB, but the reason behind this is poorly understood. To shed light on the function of HscB, we began a study on the nature of its interaction with IscU. Our work suggested that the binding site of IscU is in the C-terminal domain of HscB, and two different triple alanine substitutions ([L92A, M93A, F153A] and [E97A, E100A, E104A]) involving predicted binding site residues had detrimental effects on this interaction. However, the individual contribution of each substitution to the observed effect remains to be determined as well as the possible involvement of other residues in the proposed binding site.</p> <p>Results</p> <p>In the work reported here, we used isothermal titration calorimetry to characterize the affinity of single alanine HscB mutants for IscU, and subsequently confirmed our results with nuclear magnetic resonance spectroscopy. Alanine substitutions of L92, L96, and F153 severely impaired the ability of HscB to form a complex with IscU; substitutions of R87, R99, and E100 had more modest effects; and substitutions of T89, M93, E97, D103, E104, R152, K156, and S160 had only minor or no detectable effects.</p> <p>Conclusions</p> <p>Our results show that the residues of HscB most important for strong interaction with IscU include three hydrophobic residues (L92, L96, and F153); in addition, we identified a number of other residues whose side chains contribute to a lesser extent to the interaction. Our results suggest that the triple alanine substitution at HscB positions 92, 96, and 153 will destabilize the HscB-IscU complex by ΔΔ<it>G</it>_b≅ 5.7 kcal/mol, equivalent to a ≅ 15000-fold reduction in the affinity of HscB for IscU. We propose that this triple mutant could provide a more definitive test of the functional importance of the HscB-IscU interaction in vivo than those used previously that yielded inconclusive results.</p

A GIS-Aided Assessment of the Health Hazards of Cadmium in Farm Soils in Central Taiwan

A geostatistical method was developed to examine the correlation, or lack of it, between the levels of cadmium (Cd) detected in farm soils and those detected in the human specimens collected from residents around the contaminated areas in Changhua County where cadmium contamination of staple rice has been documented. We used the Taiwan EPA environment data in 2002 and human data which were generated by the National Health Research Institutes during 2003–2005. Kriging interpolation methods were used to determine soil Cd concentrations. A Zonal statistical function was performed to assess the individual exposure. Soil Cd levels and tissue Cd levels in residents were analyzed for contamination hotspots and other areas to determine correlation between the two variables. Three Cd contamination hotspots were identified, in which no correlation was found between soil Cd levels and tissue Cd levels in residents. Our results demonstrate how GIS spatial modeling technique can be used to estimate distribution of pollutants in an area using a limited number of data points. Results indicated no association between the soil contamination and the exposure of residents to Cd, suggesting that both the soils and the residents are receptors of Cd as a pollutant from as yet unidentified sources

High Trapped Fields in C-doped MgB2 Bulk Superconductors Fabricated by Infiltration and Growth Process.

The grain boundaries in superconducting MgB2 are known to form effective magnetic flux pinning sites and, consequently, bulk MgB2 containing a fine-grain microstructure fabricated from nanoscale Mg and B precursor powders exhibits good magnetic field-trapping performance below 20 K. We report here that the trapped field of MgB2 bulk superconductors fabricated by an infiltration and growth process to yield a dense, pore-free microstructure, can be enhanced significantly by carbon-doping, which increases intra-band scattering within the superconducting grains. A maximum trapped field of 4.15 T has been measured at 7.5 K at the centre of a five-sample stack of Mg(B1-xiCxi)2 bulk superconductors processed by infiltration and growth, which not only represents a ~40% increase in trapped field observed compared to undoped bulk MgB2, but also is the highest trapped field reported to date in MgB2 samples processed under ambient pressure. The trapped field is observed to decay at a rate of <2%/day at 10 K, which suggests that bulk MgB2 superconductors fabricated using the infiltration and growth technique can be used potentially to generate stable, high magnetic fields for a variety of engineering applications

Young Adults View Smartphone Tracking Technologies for COVID-19 as Acceptable:The Case of Taiwan

Taiwan has been successful in controlling the spread of SARS-CoV-2 during the COVID-19 pandemic; however, without a vaccine the threat of a second outbreak remains. Young adults who show few to no symptoms when infected have been identified in many countries as driving the virus' spread through unidentifiable community transmission. Mobile tracking technologies register nearby contacts of a user and notifies them if one later tests positive to the virus, potentially solving this issue; however, the effectiveness of these technologies depends on their acceptance by the public. The current study assessed attitudes towards three tracking technologies (telecommunication network tracking, a government app, and Apple and Google's Bluetooth exposure notification system) among four samples of young Taiwanese adults (aged 25 years or younger). Using Bayesian methods, we find high acceptance for all three tracking technologies (>75%), with acceptance for each technology surpassing 90% if additional privacy measures were included. We consider the policy implications of these results for Taiwan and similar cultures

Immunopanning purification and long-term culture of human retinal ganglion cells

Purpose: To establish a robust method to isolate primary retinal ganglion cells (RGCs) from human fetal retina for long-term culture while maintaining neuronal morphology and marker protein expression. Methods: A total of six human retinas were obtained from aborted fetuses at 10 to 12 weeks of gestation with informed consent from mothers. RGCs were isolated and purified by a modified two-step immunopanning procedure. The cells were maintained in a serum-free defined medium supplemented with brain-derived neurotrophic factor, ciliary neutrophic factor, and forskolin. The viable RGCs and the extent of neurite outgrowth were examined by calcein-acetoxymethylester assay. Expression of RGC markers was studied by immunocytochemistry. Results: Primary RGCs from human fetal retinas were isolated and maintained in vitro for one month with substantial neurite elongation. In cell culture, almost 70% of the isolated cells attached, spread, and displayed numerous dendrites. They were immunoreactive to RGC-specific markers (Thy-1, TUJ-1, and Brn3a) and negative for glial fibrillary acidic protein and amacrine cells marker HPC-1. Conclusions: Human RGCs were successfully isolated and maintained in long-term culture. This can serve as an ideal model for biologic, toxicological, and genomic assays of human RGCs in vitr

Sustainable Lifestyles: Practices, Policies and Barriers

A lifestyle survey is currently being conducted of Oregon residents (Spring 2015). The aim of this survey is to identify people’s attitudes, perceived social barriers, behaviors, and policies that encourage or discourage sustainable household practices as they relate to water, energy, food, transportation, and consumerism. It also examines people’s attitudes toward environmental responsibility, government regulation, and quality of life. This study, while focused on Oregon is part of a larger comparative study with Ireland. (Sources: CONSENSUS (2012); Steel (2014); Wolters, Hubbard & Steel (2010