Predictions of structural elements for the binding of Hin recombinase with the hix site of DNA

Molecular dynamics simulations were coupled with experimental data from biochemistry and genetics to generate a theoretical structure for the binding domain of Hin recombinase complexed with the hix site of DNA. The theoretical model explains the observed sequence specificity of Hin recombinase and leads to a number of testable predictions concerning altered sequence selectivity for various mutants of protein and DNA. Combining molecular dynamics simulations with constraints based on current knowledge of protein structure leads to a theoretical structure of the binding domain of Hin recombinase with the hix site of DNA. The model offers a mechanistic explanation of the presently known characteristics of Hin and predicts the effects of specific mutations of both protein and DNA. The predictions can be tested by currently feasible experiments that should lead to refinements in and improvements on the current theoretical model. Because current experimental and theoretical methods are all limited to providing only partial information about protein-DNA interactions, we believe that this approach of basing molecular simulations on experimental knowledge and using the results of these simulations to design new, more precise experimental tests will be of general utility. These results provide additional evidence for the generality of the helix-turn-helix motif in DNA recognition and stabilization of proteins on DNA

LC3 and STRAP regulate actin filament assembly by JMY during autophagosome formation.

During autophagy, actin filament networks move and remodel cellular membranes to form autophagosomes that enclose and metabolize cytoplasmic contents. Two actin regulators, WHAMM and JMY, participate in autophagosome formation, but the signals linking autophagy to actin assembly are poorly understood. We show that, in nonstarved cells, cytoplasmic JMY colocalizes with STRAP, a regulator of JMY's nuclear functions, on nonmotile vesicles with no associated actin networks. Upon starvation, JMY shifts to motile, LC3-containing membranes that move on actin comet tails. LC3 enhances JMY's de novo actin nucleation activity via a cryptic actin-binding sequence near JMY's N terminus, and STRAP inhibits JMY's ability to nucleate actin and activate the Arp2/3 complex. Cytoplasmic STRAP negatively regulates autophagy. Finally, we use purified proteins to reconstitute LC3- and JMY-dependent actin network formation on membranes and inhibition of network formation by STRAP. We conclude that LC3 and STRAP regulate JMY's actin assembly activities in trans during autophagy

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

Group Vibrational Mode Assignments as a Broadly Applicable Tool for Characterizing Ionomer Membrane Structure as a Function of Degree of Hydration

Infrared spectra of Nafion, Aquivion, and the 3M membrane were acquired during total dehydration of fully hydrated samples. Fully hydrated exchange sites are in a sulfonate form with a C₃V local symmetry. The mechanical coupling of the exchange site to a side chain ether link gives rise to vibrational group modes that are classified as C₃V modes. These mode intensities diminish concertedly with dehydration. When totally dehydrated, the sulfonic acid form of the exchange site is mechanically coupled to an ether link with no local symmetry. This gives rise to C₁ group modes that emerge at the expense of C₃V modes during dehydration. Membrane IR spectra feature a total absence of C₃V modes when totally dehydrated, overlapping C₁ and C₃V modes when partially hydrated, and a total absence of C₁ modes when fully hydrated. DFT calculated normal mode analyses complemented with molecular dynamics simulations of Nafion with overall λ (λ_(Avg)) values of 1, 3, 10, 15 and 20 waters/exchange site, were sectioned into sub-cubes to enable the manual counting of the distribution of λ_(local) values that integrate to λ_(Avg) values. This work suggests that at any state of hydration, IR spectra are a consequence of a distribution of λ_(local) values. Bond distances and the threshold value of λ_(local), for exchange site dissociation, were determined by DFT modelling and used to correlate spectra to manually counted λ_(local) distributions

On-resin N-methylation of cyclic peptides for discovery of orally bioavailable scaffolds.

Backbone N-methylation is common among peptide natural products and has a substantial impact on both the physical properties and the conformational states of cyclic peptides. However, the specific impact of N-methylation on passive membrane diffusion in cyclic peptides has not been investigated systematically. Here we report a method for the selective, on-resin N-methylation of cyclic peptides to generate compounds with drug-like membrane permeability and oral bioavailability. The selectivity and degree of N-methylation of the cyclic peptide was dependent on backbone stereochemistry, suggesting that conformation dictates the regiochemistry of the N-methylation reaction. The permeabilities of the N-methyl variants were corroborated by computational studies on a 1,024-member virtual library of N-methyl cyclic peptides. One of the most permeable compounds, a cyclic hexapeptide (molecular mass = 755 Da) with three N-methyl groups, showed an oral bioavailability of 28% in rat

Chronically Sustainable: Creating a Sustainable Community for the Chronically Ill

In a world that is rapidly changing due to climate change, there is a group of people that are being left behind in the ?green? movement. This is not new for these people; the group has often been forgotten and under served by society throughout history. This group is the chronically ill. With a variety of abilities and needs, society has failed to give them the same opportunities in life as the abled body. Buildings are required to have accessible entrances but often time they are far from the main entrances and highlight a person?s disabilities by inconveniencing not just only the disabled but everyone in their group. Yet, there is even more to accessibility than ADA entrances. Accessibility has many factors such as affordability, access to resources, and usability. This project focuses on how to create a community that is designed for the chronically ill with their various needs and abilities at the forefront of the design while considering the sustainability of the project. Located outside of Phoenix, Arizona within the Sonoran Desert, this project focuses on the ability to create a sustainable community that caters to a demographic that has struggled to become more sustainable due to their additional medical needs. The community will provide for the chronically ill and their families within the master planned community. Amenities include a community center, retail, grocery store, medical facility, athletic facilities, and leisure facilities. The project is divided into two parts: master planning the community and designing key community aspects

Proposed Modifications to the Los Angeles Rattler Test

The amount of highway construction is increasing every year. As a result, there is an ever increasing demand for mineral aggregates. Engineers classify these mineral aggregates as the smaller rocks which are composed of one or more minerals. In the state of California, between one-fifth and one-third of the 300 million dollars spent annually for the construction of highways is used for the procurement and placement of aggregates. (l) This cost is steadily increasing because of the depletion of the more accessible and higher quality aggregates. Because the traffic volume and loads are steadily increasing, the Interstate system as well as many other primary roads being built demand durable aggregates. The selecting of high quality aggregates is not a new problem. Back in Roman times it was necessary to select suitable rocks for use as flag stones, a surfacing material of the time. Some of the highways of that era remain in excellent condition today as a result of the prudent selection of surfacing material. It was not until the late nineteenth century, however, that the use of machines to simulate field conditions for the purpose of determining the durability of aggregates began. The first successful test for abrasion of aggregates was the Deval test. At the present time the Los Angeles rattler tests used by most of the State Highway Departments. It is considered the most effective test for determining the structural and abrasive qualities of an aggregate. In the late 1940\u27s however, it was observed that aggregates which had passed the Los Angeles rattler test were failing under field conditions. In 1947, Melville investigated a failure in Route 250 near Charlottesville, Virginia. From his investigation he concluded that the aggregates had weathered sufficiently to produce a layer of plastic fines which caused the failure. (2) Since then there have been many similar cases reported, especially in the western states. As a consequence of· these failures, many of the western states have been attempting to develop an empirical test that will determine the tendency of an aggregate to produce plastic fines. Washington, Oregon, Idaho, California, and South Dakota, as well as other states, have developed tests to determine the durability characteristics of aggregates. At the present time, none of these tests are universally accepted by other states or by the American Society for Testing and Materials. The Los Angeles rattler test is a very simple test and requires little time to perform. The amount of time required to conduct a test is becoming increasingly important because labor costs are steadily rising. Also, with the high production of modern aggregate contractors, it is important to have a test which can be accomplished in a short period of time. The Los Angeles rattler machine is an expensive piece of equipment and one that all state highway departments regard as standard equipment. Accordingly, this study was made with a view toward modifying the Los Angeles rattler test to include wet abrasion. This would make it possible to obtain the necessary durability information from a single laboratory test without appreciably adding to the cost of equipment

Talking about torque : measuring pack accessibility - a review

Vacuum lug closures are a simple, reliable and low-cost packaging option used for the protection and promotion of jams, pickles and sauces. Several surveys and anecdotal evidence suggest that packaging of this type can be notoriously difficult to open. Given the difficulty that packaging of this type may pose, there has been significant academic research in understanding the difficulties associated with accessing packaging of this type. In response to the qualitative data gathered in these surveys, research teams have attempted to quantify the forces that users can apply. What emerges from the approaches taken is a complex picture. Researchers do make comparisons with previous work, but numbers of people tested, materials used, diameter and posture differ between research groups as does the information and style of the dissemination of results. Future packaging research experimental design should be more thorough and consistent in the sampling and presentation of data to facilitate repeatability and validity and enable the data gathered to form a larger data set. Further, to create usable 'design limits' for manufacturers and designers to reduce the variability within the data set, more focused measurements should be taken on distinct user groups such as a specific female decile and subgroup, i.e. small-handed women between 70 and 80. Working with distinct populations would enable the likelihood of design changes to packaging to be readily compared and assessed

Strengthening and stretching for rheumatoid arthritis of the hand (SARAH). A randomised controlled trial and economic evaluation

Study registration: Current Controlled Trials ISRCTN 89936343.Background - The effectiveness of exercise for improving hand and wrist function in people with rheumatoid arthritis (RA) is uncertain. Objectives - The study aims were (1) to estimate the clinical effectiveness and cost-effectiveness of adding an optimised exercise programme for hands and upper limbs to standard care for patients with RA; and (2) to qualitatively describe the experience of participants in the trial with a particular emphasis on acceptability of the intervention, exercise behaviours and reasons for adherence/non-adherence.This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 19, No. 19. See the NIHR Journals Library website for further project information. This report has been developed in association with the NIHR Collaboration for Leadership in Applied Health Research and Care Oxford and the NIHR Biomedical Research Unit Funding Scheme. This project benefited from facilities funded through Birmingham Science City Translational Medicine Clinical Research and Infrastructure Trials Platform, with support from Advantage West Midlands

Linking the National Medical Expenditure Survey with the National Health Interview Survey: analysis of field trials

[by Nancy A. Mathiowetz and E. Patrick Ward]."September 1987."Includes bibliographical references