Biochemical and biophysical analyses of tight junction permeability made of claudin-16 and claudin-19 dimerization

The molecular nature of tight junction architecture and permeability is a long-standing mystery. Here, by comprehensive biochemical, biophysical, genetic, and electron microscopic analyses of claudin-16 and -19 interactions—two claudins that play key polygenic roles in fatal human renal disease, FHHNC—we found that 1) claudin-16 and -19 form a stable dimer through cis association of transmembrane domains 3 and 4; 2) mutations disrupting the claudin-16 and -19 cis interaction increase tight junction ultrastructural complexity but reduce tight junction permeability; and 3) no claudin hemichannel or heterotypic channel made of claudin-16 and -19 trans interaction can exist. These principles can be used to artificially alter tight junction permeabilities in various epithelia by manipulating selective claudin interactions. Our study also emphasizes the use of a novel recording approach based on scanning ion conductance microscopy to resolve tight junction permeabilities with submicrometer precision

Transient aerodynamic pressures and forces on trackside and overhead structures due to passing trains. Part 1 Model scale experiments Part 2 Standards applications

This is the first part of a two-part paper that describes the results of an experimental investigation to measure the aerodynamic pressure forces on structures in the vicinity of railway tracks. The investigations were carried out in order to obtain a fundamental understanding of the nature of the phenomenon and to obtain data for a variety of railway infrastructure geometries of particular relevance to the UK situation, in order to provide material for a National Annex to the relevant Eurocode. The experiments were carried out on the moving model TRAIN Rig, with models of three different sorts of trains with different nose types, and a variety of infrastructures types: vertical hoardings, overbridges, station canopies and trestle platforms. The transient loads that were measured had a characteristic form: a positive pressure peak followed by a negative pressure peak. In general the magnitudes of the two peaks were different, and varied with infrastructure type and position, as well as with train type. As would be expected, the more streamlined the train, the lower were the magnitudes of the pressure transients. A comparison of the experimental results was made with a variety of existing model- scale and full-scale data and a broad consistency was demonstrated, within the limits that the rather different experimental conditions in the various cases would allow. An analysis of the scaling of these pressure transients was carried out, and it was shown that whilst there was a reasonable coalescence around a theoretical formulation, the complexity of the flows involved meant that a general scaling formulation could not be achieved. Part 2 of this paper will consider the application of the results to the development of revised standards formulations. </jats:p

From Quantum Systems to L-Functions: Pair Correlation Statistics and Beyond

The discovery of connections between the distribution of energy levels of heavy nuclei and spacings between prime numbers has been one of the most surprising and fruitful observations in the twentieth century. The connection between the two areas was first observed through Montgomery's work on the pair correlation of zeros of the Riemann zeta function. As its generalizations and consequences have motivated much of the following work, and to this day remains one of the most important outstanding conjectures in the field, it occupies a central role in our discussion below. We describe some of the many techniques and results from the past sixty years, especially the important roles played by numerical and experimental investigations, that led to the discovery of the connections and progress towards understanding the behaviors. In our survey of these two areas, we describe the common mathematics that explains the remarkable universality. We conclude with some thoughts on what might lie ahead in the pair correlation of zeros of the zeta function, and other similar quantities.Comment: Version 1.1, 50 pages, 6 figures. To appear in "Open Problems in Mathematics", Editors John Nash and Michael Th. Rassias. arXiv admin note: text overlap with arXiv:0909.491

Mutational analyses of UPIIIA, SHH, EFNB2, and HNF1β in persistent cloaca and associated kidney malformations

OBJECTIVES: ‘Persistent cloaca’ is a severe malformation affecting females in which the urinary, genital and alimentary tracts share a single conduit. Previously, a Uroplakin IIIA (UPIIIA) mutation was reported in one individual with persistent cloaca, and UPIIIA, Sonic Hedgehog (SHH), Ephrin B2 (EFNB2) and Hepatocyte Nuclear Factor 1β (HNF1β) are expressed during the normal development of organs that are affected in this condition. HNF1β mutations have been associated with uterine malformations in humans, and mutations of genes homologous to human SHH or EFNB2 cause persistent cloaca in mice. PATIENTS AND METHODS: We sought mutations of coding regions of UPIIIA, SHH, EFNB2 and HNF1β genes by direct sequencing in a group of 20 patients with persistent cloaca. Most had associated malformations of the upper renal tract and over half had impaired renal excretory function. The majority of patients had congenital anomalies outside the renal/genital tracts and two had the VACTERL association. RESULTS: Apart from a previously described index case, we failed to find UPIIIA mutations, and no patient had a SHH, EFNB2 or HNF1β mutation. CONCLUSION: Persistent cloaca is only rarely associated with UPIIIA mutation. Despite the fact that SHH and EFNB2 are appealing candidate genes, based on their expression patterns and mutant mice phenotypes, they were not mutated in these humans with persistent cloaca. Although HNF1β mutations can perturb paramesonephric duct fusion in humans, HNF1β was not mutated in persistent cloaca

Asymmetric triplex metallohelices with high and selective activity against cancer cells

Small cationic amphiphilic α-helical peptides are emerging as agents for the treatment of cancer and infection, but they are costly and display unfavourable pharmacokinetics. Helical coordination complexes may offer a three-dimensional scaffold for the synthesis of mimetic architectures. However, the high symmetry and modest functionality of current systems offer little scope to tailor the structure to interact with specific biomolecular targets, or to create libraries for phenotypic screens. Here, we report the highly stereoselective asymmetric self-assembly of very stable, functionalized metallohelices. Their anti-parallel head-to-head-to-tail ‘triplex’ strand arrangement creates an amphipathic functional topology akin to that of the active sub-units of, for example, host-defence peptides and ​p53. The metallohelices display high, structure-dependent toxicity to the human colon carcinoma cell-line HCT116 ​p53++, causing dramatic changes in the cell cycle without DNA damage. They have lower toxicity to human breast adenocarcinoma cells (MDA-MB-468) and, most remarkably, they show no significant toxicity to the bacteria methicillin-resistant Staphylococcus aureus and Escherichia coli. At a glanc

A Threat Assessment and Security Analysis of the Three Sports Facilities of Indiana University-Purdue University, Indianapolis NCAA Softball Fields, Carroll Stadium, and the IU Natatorium

This research report provides a security assessment of the Softball Fields, Carroll Stadium, and the Natatorium Complex at Indiana University-Purdue University Indianapolis (IUPUI). The purpose of this report is to prevent and mitigate harm to visitors and these facilities which resulting from human-made or natural disasters. Research is guided by the hypothesis that these facilities- due to their respective importance, locations, and attendance patterns are in harm’s way; and that certain strategies of prevention, protection, and mitigation coupled with effective preparedness, response, and recovery can lessen risk, improve security and provide A THREAT ASSESSMENT AND SECURITY ANALYSIS 5 added resilience. Further, “harm’s way” is considered to be either a natural disaster or a human-made disaster, accident, active provocation, or act of terrorism. Methods of analysis include applied research; predominantly utilizing qualitative data with some quantitative investigation. Results of this assessment illustrate that these venues possess numerous vulnerabilities to both natural and human-made threats that if exposed, could result in serious consequences. The two most likely natural hazards identified include straight-line winds and tornadoes. Further, the most likely human threats to these facilities arise from a potential terrorist vehicle attack (TVA) and an active shooter. This project also identifies a specific need for additional planning to prevent an IED or VBIED attack on the Natatorium. Common themes from the attached three case studies reveal that given theses vulnerabilities, the following safety and security adjustments are recommended: Surveillance equipment Metal detectors Security bollards or other temporary barriers Evacuation routes and shelter in place plans Special event security procedures Weather related technology and protocols Staff training for emergency situation

Multi-centre parallel arm randomised controlled trial to assess the effectiveness and cost-effectiveness of a group-based cognitive behavioural approach to managing fatigue in people with multiple sclerosis

Abstract (provisional) Background Fatigue is one of the most commonly reported and debilitating symptoms of multiple sclerosis (MS); approximately two-thirds of people with MS consider it to be one of their three most troubling symptoms. It may limit or prevent participation in everyday activities, work, leisure, and social pursuits, reduce psychological well-being and is one of the key precipitants of early retirement. Energy effectiveness approaches have been shown to be effective in reducing MS-fatigue, increasing self-efficacy and improving quality of life. Cognitive behavioural approaches have been found to be effective for managing fatigue in other conditions, such as chronic fatigue syndrome, and more recently, in MS. The aim of this pragmatic trial is to evaluate the clinical and cost-effectiveness of a recently developed group-based fatigue management intervention (that blends cognitive behavioural and energy effectiveness approaches) compared with current local practice. Methods This is a multi-centre parallel arm block-randomised controlled trial (RCT) of a six session group-based fatigue management intervention, delivered by health professionals, compared with current local practice. 180 consenting adults with a confirmed diagnosis of MS and significant fatigue levels, recruited via secondary/primary care or newsletters/websites, will be randomised to receive the fatigue management intervention or current local practice. An economic evaluation will be undertaken alongside the trial. Primary outcomes are fatigue severity, self-efficacy and disease-specific quality of life. Secondary outcomes include fatigue impact, general quality of life, mood, activity patterns, and cost-effectiveness. Outcomes in those receiving the fatigue management intervention will be measured 1 week prior to, and 1, 4, and 12 months after the intervention (and at equivalent times in those receiving current local practice). A qualitative component will examine what aspects of the fatigue management intervention participants found helpful/unhelpful and barriers to change. Discussion This trial is the fourth stage of a research programme that has followed the Medical Research Council guidance for developing and evaluating complex interventions. What makes the intervention unique is that it blends cognitive behavioural and energy effectiveness approaches. A potential strength of the intervention is that it could be integrated into existing service delivery models as it has been designed to be delivered by staff already working with people with MS. Service users will be involved throughout this research. Trial registration: Current Controlled Trials ISRCTN7651747

Ovine pedomics : the first study of the ovine foot 16S rRNA-based microbiome

We report the first study of the bacterial microbiome of ovine interdigital skin based on 16S rRNA by pyrosequencing and conventional cloning with Sanger-sequencing. Three flocks were selected, one a flock with no signs of footrot or interdigital dermatitis, a second flock with interdigital dermatitis alone and a third flock with both interdigital dermatitis and footrot. The sheep were classified as having either healthy interdigital skin (H), interdigital dermatitis (ID) or virulent footrot (VFR). The ovine interdigital skin bacterial community varied significantly by flock and clinical condition. The diversity and richness of operational taxonomic units was greater in tissue from sheep with ID than H or VFR affected sheep. Actinobacteria, Bacteriodetes, Firmicutes and Proteobacteria were the most abundant phyla comprising 25 genera. Peptostreptococcus, Corynebacterium and Staphylococcus were associated with H, ID and VFR respectively. Sequences of Dichelobacter nodosus, the causal agent of ovine footrot, were not amplified due to mismatches in the 16S rRNA universal forward primer (27F). A specific real time PCR assay was used to demonstrate the presence of D. nodosus which was detected in all samples including the flock with no signs of ID or VFR. Sheep with ID had significantly higher numbers of D. nodosus (104-109 cells/g tissue) than those with H or VFR feet