The alternating access mechanism of transport as observed in the sodium-hydantoin transporter Mhp1

Crystal structures of a membrane protein transporter in three different conformational states provide insights into the transport mechanism

Lower limb stiffness and maximal sprint speed in 11-16-year-old boys

The purpose of the study was to examine the relationship between vertical stiffness, leg stiffness and maximal sprint speed in a large cohort of 11-16-year-old boys. Three-hundred and thirty-six boys undertook a 30 m sprint test using a floor-level optical measurement system, positioned in the final 15 m section. Measures of speed, step length, step frequency, contact time and flight time were directly measured whilst force, displacement, vertical stiffness and leg stiffness, were modeled from contact and flight times, from the two fastest consecutive steps for each participant over two trials. All force, displacement and stiffness variables were significantly correlated with maximal sprint speed (p 0.7) relationship with sprint speed, while vertical center of mass displacement, absolute vertical stiffness, relative peak force, and maximal leg spring displacement had large (r > 0.5) relationships. Relative vertical stiffness and relative peak force did not significantly change with advancing age (p > 0.05), but together with maximal leg spring displacement accounted for 96% of the variance in maximal speed. It appears that relative vertical stiffness and relative peak force are important determinants of sprint speed in boys aged 11-16 years, but are qualities that may need to be trained due to no apparent increases from natural development. Practitioners may wish to utilize training modalities such as plyometrics and resistance training to enable adaptation to these qualities due to their importance as predictors of speed in youth

Molecular mechanism of ligand recognition by membrane transport protein, Mhp1

The hydantoin transporter Mhp1 is a sodium-coupled secondary active transport protein of the nucleobase-cation-symport family and a member of the widespread 5-helix inverted repeat superfamily of transporters. The structure of Mhp1 was previously solved in three different conformations providing insight into the molecular basis of the alternating access mechanism. Here, we elucidate detailed events of substrate binding, through a combination of crystallography, molecular dynamics, site-directed mutagenesis, biochemical/biophysical assays, and the design and synthesis of novel ligands. We show precisely where 5-substituted hydantoin substrates bind in an extended configuration at the interface of the bundle and hash domains. They are recognised through hydrogen bonds to the hydantoin moiety and the complementarity of the 5-substituent for a hydrophobic pocket in the protein. Furthermore, we describe a novel structure of an intermediate state of the protein with the external thin gate locked open by an inhibitor, 5-(2-naphthylmethyl)-L-hydantoin, which becomes a substrate when leucine 363 is changed to an alanine. We deduce the molecular events that underlie acquisition and transport of a ligand by Mhp1

A New Direction to Athletic Performance: Understanding the Acute and Longitudinal Responses to Backward Running

Backward running (BR) is a form of locomotion that occurs in short bursts during many overground field and court sports. It has also traditionally been used in clinical settings as a method to rehabilitate lower body injuries. Comparisons between BR and forward running (FR) have led to the discovery that both may be generated by the same neural circuitry. Comparisons of the acute responses to FR reveal that BR is characterised by a smaller ratio of braking to propulsive forces, increased step frequency, decreased step length, increased muscle activity and reliance on isometric and concentric muscle actions. These biomechanical differences have been critical in informing recent scientific explorations which have discovered that BR can be used as a method for reducing injury and improving a variety of physical attributes deemed advantageous to sports performance. This includes improved lower body strength and power, decreased injury prevalence and improvements in change of direction performance following BR training. The current findings from research help improve our understanding of BR biomechanics and provide evidence which supports BR as a useful method to improve athlete performance. However, further acute and longitudinal research is needed to better understand the utility of BR in athletic performance programs

Right drug, right patient, right time: aspiration or future promise for biologics in rheumatoid arthritis?

Individualising biologic disease-modifying anti-rheumatic drugs (bDMARDs) to maximise outcomes and deliver safe and cost-effective care is a key goal in the management of rheumatoid arthritis (RA). Investigation to identify predictive tools of bDMARD response is a highly active and prolific area of research. In addition to clinical phenotyping, cellular and molecular characterisation of synovial tissue and blood in patients with RA, using different technologies, can facilitate predictive testing. This narrative review will summarise the literature for the available bDMARD classes and focus on where progress has been made. We will also look ahead and consider the increasing use of ‘omics’ technologies, the potential they hold as well as the challenges, and what is needed in the future to fully realise our ambition of personalised bDMARD treatment

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

ansa-metallocene derivatives : XXV. Synthesis, crystal structure and reactions of a tetramethylethano-bridged vanadocene dichloride, (CH₃)₄C₂(C₅H₄)₂VCl₂

The tetramethylethano-bridged vanadocene complex (CH3)4C2(C5H4)2VCl2 has been made in two ways: (i) from VCl3·3THF by reaction with (CH3)4C2(C5H4MgCl·2THF)2 in THF and subsequent oxidation with PCl3, and (ii) from VCl4·2THF by reaction with (CH3)4C2(C5H4Li)2 or with (CH3)4C2(C5H4MgCl·2THF)2 in a toluene/dimethoxyethane mixture. Its crystal structure is in accordance with expectations. Reduction of (CH3)4C2(C5H4)2VCl2 with tetrakis(dimethylamino)ethene in toluene or with LiAlH4 in THF yields the monochloride (CH3)4C2(C5H4)2VCl; from this VIII compound cationic complexes (CH3)4C2(C5H4)2VL2+ with L = CO or CNtBu are readily obtained by ligand exchange. 1H NMR signals of these diamagnetic complexes have been assigned to H atoms in α and β C5-ring positions by use of selective nuclear Overhauser effects. The ring-bridged VII complex (CH3)4C2(C5H4)2V could not be prepared. Cyclovoltammetry studies indicate that (in contrast to its unbridged counterpart (C5H5)2V) this VII complex decays within seconds after its formation by electrochemical reduction of its chloro derivatives