Patellar tendon in vivo regional strain with varying knee angle

Purpose: Descriptive data on the aspects of site specific in vivo tendon strain with varying knee joint angle are non-existent. The present study determines and compares surface and deep layer strain of the patellar tendon during isometric contractions across a range of knee joint angles. Methods: Male participants (age 22.0 ± 3.4) performed ramped isometric knee extensions at knee joint angles of 900, 700, 500 and 300 of flexion. Strain patterns of the anterior and posterior regions of the patellar tendon were determined using real-time B-mode ultrasonography at each knee joint angle. Regional strain measures were compared using an automated pixel tracking method. Results: Strain was seen to be greatest for both the anterior and posterior regions with the knee at 900 (7.76 ± 0.89% and 5.06 ± 0.76%). Anterior strain was seen to be significantly greater (p<0.05) than posterior strain for all knee angles apart from 300, 900 = (7.76 vs. 5.06%), 700 = (4.77 vs. 3.75%), and 500 = (3.74 vs. 2.90%). The relative strain (ratio of anterior to posterior), was greatest with the knee joint angle at 900, and decreased as the knee joint angle reduced. Conclusions: The results from this study indicate that not only are there greater absolute tendon strains with the knee in greater flexion, but that the knee joint angle affects the regional strain differentially, resulting in greater shear between the tendon layers with force application when the knee is in greater degrees of flexion. These results have important implications for rehabilitation and training

Sequential and counter-selectable cassettes for fission yeast

BACKGROUND: Fission yeast is one of the most commonly used model organisms for studying genetics. For selection of desirable genotypes, antibiotic resistance cassettes are widely integrated into the genome near genes of interest. In yeasts, this is achieved by PCR amplification of the cassette flanked by short homology sequences, which can be incorporated by homology directed repair. However, the currently available cassettes all share the same tef promoter and terminator sequences. It can therefore be challenging to perform multiple genetic modifications by PCR-based targeting, as existing resistance cassettes in strains can be favored for recombination due to shared homology between the cassettes. RESULTS: Here we have generated new selection cassettes that do not recombine with those traditionally used. We achieved this by swapping the tef promoter and terminator sequences in the established antibiotic resistance MX6 cassette series for alternative promoters and/or terminators. The newly created selection cassettes did not recombine with the tef-containing MX6 cassettes already present in the genome, allowing for sequential gene targeting using the PCR-based method. In addition, we have generated a series of plasmids to facilitate the C-terminal tagging of genes with desired epitopes. We also utilized the anti-selection gene HSV-TK, which results in cell death in strains grown on the drug 5-Fluoro-2'-deoxyuridine (FdU, Floxuridin or FUDR). By fusing an antibiotic resistance gene to HSV-TK, we were able to select on the relevant antibiotic as well as counter-select on FdU media to confirm the desired genomic modification had been made. We noted that the efficiency of the counter selection by FdU was enhanced by treatment with hydroxyurea. However, a number of DNA replication checkpoint and homologous recombination mutants, including rad3∆, cds1∆, rad54∆ and rad55∆, exhibited sensitivity to FdU even though those strains did not carry the HSV-TK gene. To remove counter-selectable markers, we introduced the Cre-loxP irreversible recombination method. Finally, utilizing the negative selectable markers, we showed efficient induction of point mutations in an endogenous gene by a two-step transformation method. CONCLUSIONS: The plasmid constructs and techniques described here are invaluable tools for sequential gene targeting and will simplify construction of fission yeast strains required for study

Analytical Exploration of Potential Pathways by which Diabetes Mellitus Impacts Tuberculosis Epidemiology.

We aimed to develop a conceptual framework of diabetes mellitus (DM) effects on tuberculosis (TB) natural history and treatment outcomes, and to assess the impact of these effects on TB-transmission dynamics. The model was calibrated using TB data for India. A conceptual framework was developed based on a literature review, and then translated into a mathematical model to assess the impact of the DM-on-TB effects. The impact was analyzed using TB-disease incidence hazard ratio (HR) and population attributable fraction (PAF) measures. Evidence was identified for 10 plausible DM-on-TB effects. Assuming a flat change of 300% (meaning an effect size of 3.0) for each DM-on-TB effect, the HR ranged between 1.0 (Effect 9-Recovery) and 2.7 (Effect 2-Fast progression); most effects did not have an impact on the HR. Meanwhile, TB-disease incidence attributed directly and indirectly to each effect ranged between -4.6% (Effect 7-TB mortality) and 34.5% (Effect 2-Fast progression). The second largest impact was for Effect 6-Disease infectiousness at 29.9%. In conclusion, DM can affect TB-transmission dynamics in multiple ways, most of which are poorly characterized and difficult to assess in epidemiologic studies. The indirect (e.g. onward transmission) impacts of some DM-on-TB effects are comparable in scale to the direct impacts. While the impact of several effects on the HR was limited, the impact on the PAF was substantial suggesting that DM could be impacting TB epidemiology to a larger extent than previously thought

Towards Emotion Recognition: A Persistent Entropy Application

Emotion recognition and classification is a very active area of research. In this paper, we present a first approach to emotion classification using persistent entropy and support vector machines. A topology-based model is applied to obtain a single real number from each raw signal. These data are used as input of a support vector machine to classify signals into 8 different emotions (calm, happy, sad, angry, fearful, disgust and surprised)

LARP7 family proteins have conserved function in telomerase assembly

Understanding the intricacies of telomerase regulation is crucial due to the potential health benefits of modifying its activity. Telomerase is composed of an RNA component and reverse transcriptase. However, additional factors required during biogenesis vary between species. Here we have identified fission yeast Lar7 as a member of the conserved LARP7 family, which includes the Tetrahymena telomerase-binding protein p65 and human LARP7. We show that Lar7 has conserved RNA-recognition motifs, which bind telomerase RNA to protect it from exosomal degradation. In addition, Lar7 is required to stabilise the association of telomerase RNA with the protective complex LSm2-8, and telomerase reverse transcriptase. Lar7 remains a component of the mature telomerase complex and is required for telomerase localisation to the telomere. Collectively, we demonstrate that Lar7 is a crucial player in fission yeast telomerase biogenesis, similarly to p65 in Tetrahymena, and highlight the LARP7 family as a conserved factor in telomere maintenance

Telomere protein Rap1 is a charge resistant scaffolding protein in chromosomal bouquet formation

© 2015 Amelina et al. Background: Chromosomes reorganize in early meiotic prophase to form the so-called telomere bouquet. In fission yeast, telomeres localize to the nuclear periphery via interaction of the telomeric protein Rap1 with the membrane protein Bqt4. During meiotic prophase, the meiotic proteins Bqt1-2 bind Rap1 and tether to the spindle pole body to form the bouquet. Although it is known that this polarized chromosomal arrangement plays a crucial role in meiotic progression, the molecular mechanisms of telomere bouquet regulation are poorly understood. Results: Here, we detected high levels of Rap1 phospho-modification throughout meiotic prophase, and identified a maximum of 35 phosphorylation sites. Concomitant phosphomimetic mutation of the modification sites suggests that Rap1 hyper-phosphorylation does not directly regulate telomere bouquet formation or dissociation. Despite the negative charge conferred by its highly phosphorylated state, Rap1 maintains interactions with its binding partners. Interestingly, mutations that change the charge of negatively charged residues within the Bqt1-2 binding site of Rap1 abolished the affinity to the Bqt1-2 complex, suggesting that the intrinsic negative charge of Rap1 is crucial for telomere bouquet formation. Conclusions: Whereas Rap1 hyper-phosphorylation observed in meiotic prophase does not have an apparent role in bouquet formation, the intrinsic negative charge of Rap1 is important for forming interactions with its binding partners. Thus, Rap1 is able to retain bouquet formation under heavily phosphorylated status.This work is supported mainly by the European Research Council (281722-HRMCB) and partly by Cancer Research UK (C36439/A12097) and the Cancer Research UK - UCL Centre

The role of temperature and frequency on fretting wear of a like-on-like stainless steel contact

The influences of environmental temperature and fretting frequency on the mechanisms and rates of wear in a like-on-like 304 stainless steel contact were examined, and mainly attributed to changes in the mechanical response of the bulk material and to changes in the behaviour of the oxide debris formed in the fretting process. At low temperatures, wear proceeds by continual oxide formation and egress from the contact, whilst at high temperatures, the rate of wear is much reduced, associated with the development of oxide formed into a protective bed within the contact. The temperature at which the change between these two behaviours took place was dependent upon the fretting frequency, with evidence that, at this transition temperature, changes in behaviour can occur as the fretting test proceeds under a fixed set of conditions. An interaction diagram has been developed which provides a coherent framework by which the complex effects of these two parameters can be rationalised in terms of widely accepted physical principles

Risk stratification by pre-operative cardiopulmonary exercise testing improves outcomes following elective abdominal aortic aneurysm surgery : a cohort study

Background: In 2009, the NHS evidence adoption center and National Institute for Health and Care Excellence (NICE) published a review of the use of endovascular aneurysm repair (EVAR) of abdominal aortic aneurysms (AAAs). They recommended the development of a risk-assessment tool to help identify AAA patients with greater or lesser risk of operative mortality and to contribute to mortality prediction. A low anaerobic threshold (AT), which is a reliable, objective measure of pre-operative cardiorespiratory fitness, as determined by pre-operative cardiopulmonary exercise testing (CPET) is associated with poor surgical outcomes for major abdominal surgery. We aimed to assess the impact of a CPET-based risk-stratification strategy upon perioperative mortality, length of stay and non-operative costs for elective (open and endovascular) infra-renal AAA patients. Methods: A retrospective cohort study was undertaken. Pre-operative CPET-based selection for elective surgical intervention was introduced in 2007. An anonymized cohort of 230 consecutive infra-renal AAA patients (2007 to 2011) was studied. A historical control group of 128 consecutive infra-renal AAA patients (2003 to 2007) was identified for comparison. Comparative analysis of demographic and outcome data for CPET-pass (AT ≥ 11 ml/kg/min), CPET-fail (AT < 11 ml/kg/min) and CPET-submaximal (no AT generated) subgroups with control subjects was performed. Primary outcomes included 30-day mortality, survival and length of stay (LOS); secondary outcomes were non-operative inpatient costs. Results: Of 230 subjects, 188 underwent CPET: CPET-pass n = 131, CPET-fail n = 35 and CPET-submaximal n = 22. When compared to the controls, CPET-pass patients exhibited reduced median total LOS (10 vs 13 days for open surgery, n = 74, P < 0.01 and 4 vs 6 days for EVAR, n = 29, P < 0.05), intensive therapy unit requirement (3 vs 4 days for open repair only, P < 0.001), non-operative costs (£5,387 vs £9,634 for open repair, P < 0.001) and perioperative mortality (2.7% vs 12.6% (odds ratio: 0.19) for open repair only, P < 0.05). CPET-stratified (open/endovascular) patients exhibited a mid-term survival benefit (P < 0.05). Conclusion: In this retrospective cohort study, a pre-operative AT > 11 ml/kg/min was associated with reduced perioperative mortality (open cases only), LOS, survival and inpatient costs (open and endovascular repair) for elective infra-renal AAA surgery

Dynamics of temporally interleaved percept-choice sequences: interaction via adaptation in shared neural populations

At the onset of visually ambiguous or conflicting stimuli, our visual system quickly ‘chooses’ one of the possible percepts. Interrupted presentation of the same stimuli has revealed that each percept-choice depends strongly on the history of previous choices and the duration of the interruptions. Recent psychophysics and modeling has discovered increasingly rich dynamical structure in such percept-choice sequences, and explained or predicted these patterns in terms of simple neural mechanisms: fast cross-inhibition and slow shunting adaptation that also causes a near-threshold facilitatory effect. However, we still lack a clear understanding of the dynamical interactions between two distinct, temporally interleaved, percept-choice sequences—a type of experiment that probes which feature-level neural network connectivity and dynamics allow the visual system to resolve the vast ambiguity of everyday vision. Here, we fill this gap. We first show that a simple column-structured neural network captures the known phenomenology, and then identify and analyze the crucial underlying mechanism via two stages of model-reduction: A 6-population reduction shows how temporally well-separated sequences become coupled via adaptation in neurons that are shared between the populations driven by either of the two sequences. The essential dynamics can then be reduced further, to a set of iterated adaptation-maps. This enables detailed analysis, resulting in the prediction of phase-diagrams of possible sequence-pair patterns and their response to perturbations. These predictions invite a variety of future experiments