Mccarthythestrup2801

[Purpose] To investigate the interaction between locomotion and improvements in performing self-care. [Participants and Methods] We retrospectively analyzed 930 patients with stroke who were registered in the Japanese Rehabilitation Database. We performed a correlation analysis to evaluate the relationships among all the collected data. Then, hierarchical multiple regression analysis was performed using the self-care motor score of the Functional Independent Measure (FIM) as the dependent variable. "Model 1" used two independent variables (National Institute of Health Stroke and Rankin Scale), "model 2" used two independent variables (locomotion gain and gain of an item with the closest coefficient correlation added to model 1), and "model 3" used a mean-centering value, which was added to model 2. R2 values were calculated using a simple slope analysis. [Results] Locomotion showed an interaction with three self-care activities. The R2 changes in models 1 and 2 (ΔR2) were significant for dressing upper body (ΔR2=0.001), bowel management (ΔR2=0.006), and toileting (ΔR2=0.006). The results of the simple slope analysis were significant. [Conclusion] Locomotion demonstrated an interaction with various activities for improving self-care. There were varying degrees of improvement in self-care despite a uniform improvement in the degree of locomotion. Therefore, locomotion interaction should be considered for each intervention that targets activities of daily living.[Purpose] This study evaluated subjective posture recognition by physiotherapists with expertise in posture, examined the quantification of posture using a three-dimensional (3D) motion capture, and described posture-based characteristics. [Participants and Methods] We photographed good, normal, and bad postures in 12 participants using an infrared camera, and the resultant data were analyzed. [Results] We observed the largest displacement from a good to a bad posture in the tenth thoracic vertebra on the X-axis in the anterior-posterior direction in comparison with other index points. Further, we observed considerable differences between good and bad postures compared with other index points. Moreover, we noted significant differences between the amount of displacement between good to a normal posture and from a good to a bad posture. The vertical displacement of the Z-axis was smaller than other index points. [Conclusion] Th10 captured features from the three postures. The X-axis was displaced most between good and bad postures. Further, the amount of displacement on the Z-axis was less between good and bad posture, rendering it difficult to capture features. Therefore, the findings reported herein can be used to compare the front and rear directions of the X-axis for capturing postural changes.[Purpose] This study aimed to examine the thermal skin responses (thermal buildup and retention rate) to instrument-assisted soft tissue mobilization (IASTM) procedures applied on hamstrings at different angles. [Participants and Methods] Thirty university students (age 20 ± 4 years, weight 70.61 ± 9.11 kg, height 168.5 ± 7.5 cm) received three sessions of 10-min Ergon® IASTM treatment on their dominant limbs' hamstrings at 20°, 60°, and 90° application angles, respectively. The skin temperature was measured with a thermometer immediately before and after treatment, and every minute thereafter until it returned to the baseline value. [Results] IASTM resulted in a significant increase in skin temperature irrespective of the application angle. The thermal retention rate produced by the treatment at a 90° angle was significantly higher than that produced by the 20° application angle (78.9 vs. 64.53 min). No significant differences were observed between the 60° and 90° angle applications (72.5 vs. 78.9 min). [Conclusion] IASTM application at 60° and 90° angles can increase and retain the hamstring's skin temperature for more than an hour, creating the conditions for potential positive adaptations to local metabolism and muscle tone.. [Purpose] Arm swing is seldom considered while designing clinical rehabilitation protocols for hemiplegic patients with upper or lower extremity disabilities, likely due to the unclear role that arm swinging plays in the ability to walk. We, therefore, aimed to investigate the effect of arm swinging on walking abilities. [Participants and Methods] The study enrolled 20 healthy adults who performed a 10 m walking test with normal gait, single-arm restricted gait, both-arms restricted gait, and maximum arm-swing gait with one arm fixed in the Wernicke-Mann's position. The walking time, number of steps taken, and pelvic fluctuation were measured for the four gaits. A fixed-trunk type arm sling was used for maintaining the Wernicke-Mann's position. [Results] Velocity and stride length decreased significantly while walking with the single-arm restricted gait and both-arms restricted gait in comparison to normal gait. The maximum arm-swing gait showed no significant differences from normal gait in terms of cadence, velocity, and stride. Pelvic fluctuations also had no significant differences among all gaits. [Conclusion] Restricting movement of one or both arms limited the walking speed and stride; however, in Wernicke-Mann's limb position, if the arm is intentionally swung, the walking speed and stride resembled that of normal gait.[Purpose] This study investigated the effects of manual manipulation therapy on the pain and dysfunction of patients with lumbar spinal stenosis. [Participants and Methods] In this study, 30 patients with chronic back pain were evenly divided into an experimental group, who received manual traction therapy, and a control group, who received intermittent traction therapy. Both groups received therapy three times a week for eight weeks. A visual analogue scale was used to measure participants' back pain, and the Oswestry disability index (ODI) was used to check the functional impediment they experienced as a result. [Results] The intragroup comparison showed that the visual analog scale and the ODI significantly decreased in the control group and the experimental group, respectively. The intergroup comparison after treatment showed that the visual analog scale and the ODI of the experimental group were significantly lower than in the control group. [Conclusion] The results of this study suggest that manual manipulation therapy is an effective intervention for treating pain and dysfunction in patients with lumbar spinal stenosis.[Purpose] The purpose of this study is to consider the correlation between ankle plantar flexor strength and leg extensor torque in order to investigate whether the leg extension torque can be expected to increase as the triceps surae muscle strength is increased. [Participants and Methods] Healthy adults of 30 males and 22 females were recruited. Hand Held Dynamometer was used to measure ankle plantar flexor strength. Strength Ergo 240 was used to measure leg extensor torque. After measurement, a correlation between these factors was investigated by gender. [Results] For both males and females, a significant positive correlation between the left and right ankle plantar flexor strength and leg extensor torque was observed. [Conclusion] Actively performing muscle strengthening exercises for ankle plantar flexor by physical therapists was found to be meaningful in increasing leg extensor torque.[Purpose] The purpose of this study was to assess the effects of flexi bar training model and moderate running exercise on health-related physical fitness in overweight adults. [Participants and Methods] Forty participants were randomly assigned to an experimental (20 participant performing flexi bar training model (FBT)) and control (20 participant performing moderate running exercise (MRE) group. The participant in both groups then underwent program training 50 min/day, 3 times a week, for 12 weeks. The main outcome measures were health related physical fitness (HRPF). [Results] The result showed significant differences between FBT and MRE group. After 12 weeks FBT showed improve HRPF variable. [Conclusion] flexi bar training model can improvement health related physical fitness in overweight adults.[Purpose] The purpose of this longitudinal study was to investigate the diversity in infant crawling and examine the quantitative regularity in crawling variations necessary for the acquisition of walking in infants with typical development. [Participants and Methods] Infants with no neurological or orthopedic problems participated in this study. Using Internet Protocol (IP) cameras, crawling was simultaneously filmed from six different angles. Filming was continued until the acquisition of independent walking. The crawling movement in the video was coded. We considered the number of different completed codes as the number of variations and examined the cumulative number during the filming period in each participant. [Results] Nineteen infants completed the study. The pattern of change in the cumulative number of variations with increasing age (in days) varied between cases. Although the cumulative number of crawling variations at the time of acquisition of independent walking was inconsistent, it was negatively correlated with the crawling start age (in days). [Conclusion] Diversity exists in infant crawling. Infants who start crawling at a younger age tend to express more variation, whereas infants who start crawling when older tend to express less variation.In many trials, the duration between patient enrolment and an event occurring is used as the efficacy endpoint. Common endpoints of this type include the time until relapse, progression to the next stage of a disease, or time until remission. The criteria of an event may be defined by multiple components, one or more of which may be a continuous measurement being above or below a threshold. Typical analyses consider all components as binary variables and record the first time at which the patient has an event. This is analysed through constructing and testing survival functions using Kaplan-Meier, parametric models or Cox models. This approach ignores information contained in the continuous components. We propose a method that makes use of this information to improve the precision of analyses using these types of endpoints. We use joint modelling of the continuous and binary components to construct survival curves. We show how to compute confidence intervals for quantities of interest, such as the median or mean event time. We assess the properties of the proposed method using simulations and data from a phase II cancer trial and an observational study in renal disease.Tropical forests are a critical component of the Earth system, storing half of the global forest carbon stocks and accounting for a third of terrestrial photosynthesis. Lianas are structural parasites that can substantially reduce the carbon sequestration capacity of these forests. Simulations of this peculiar growth form have only recently started and a single vegetation model included lianas so far. In this work we present a new liana implementation within the individual based model Formind. Initial tests indicate high structural realism both horizontal and vertical. In particular, we benchmarked the model against empirical observations of size distribution, mean liana cluster size and vertical leaf distribution for the Paracou site in French Guiana. Our model predicted a reduction of above-ground biomass between 10% for mature stands to 45% for secondary plots upon inclusion of lianas in the simulations. The reduced biomass was the result of a lower productivity due to a combination of lower tree photosynthesis and high liana respiration. We evaluated structural metrics (LAI, basal area, mean tree-height) and carbon fluxes (GPP, respiration) by comparing simulations with and without lianas. At the equilibrium, liana productivity was 1.9t C ha - 1 y - 1 , or 23% of the total GPP and the forest carbon stocks were between 5% and 11% lower in simulations with lianas. We also highlight the main strengths and limitations of this new approach and propose new field measurements to further the understanding of liana ecology in a modelling framework.Under the banner of a "New Green Revolution for Africa," agricultural intensification programs aim to make smallholder agriculture more productive as well as "climate smart". As with Green Revolutions in Asia and Mexico, agricultural innovations (hybrid seeds, agronomic engineering, market linkages,and increased use of fertilizer and pesticides) are promoted as essential catalysts of agriculture-led economic growth. Intensification programs are now frequently linked to Climate Smart Agriculture (CSA), which attempts to build resilience and reduce greenhouse gas emissions while increasing crop yields. This article considers who and what is resilient in Africa's Green Revolution. We report on a multi-season study of smallholder food producers' experiences with Rwanda's Crop Intensification Program (CIP) and related policies that aim to commercialize subsistence agriculture while implementing CSA. . We suggest that there are fundamental limits to the climate resilience afforded by CSA and development efforts rooted in Green Revolution thinking. Our findings illustrate that such efforts foreground technology and management adjustments in ways that have reduced smallholder resilience by inhibiting sovereignty over land use, decreasing livelihood flexibility, and constricting resource access. We put forth that rural development policies could better promote climate-resilient livelihoods through 1) adaptive governance that enables smallholder land use decision-making; 2) support for smallholder food producers' existing agro-ecological strategies of intensification; 3) participatory approaches to visualize and correct for inequalities in local processes of social-ecological resilence Such considerations are paramount for meeting the United Nations Sustainable Development Goals and building climate-resilient food systems.A highly diastereoselective three-component C-H bond addition across butadiene and activated ketones is described. This transformation provides homoallylic tertiary alcohols through the formation of two C-C σ bonds and with complete selectivity for an E-alkene isomer. The reaction exhibits good scope with respect to activated ketone inputs, including highly strained cyclic and electron-deficient cyclic and acyclic ketones. Additionally, high diastereoselectivities were achieved for alcohols prepared from unsymmetrical ketones.The increasing impact of humans on land and ongoing global population growth requires an improved understanding of land cover (LC) and land use (LU) processes related to settlements. The heterogeneity of built-up areas and infrastructures as well as the importance of not only mapping, but also characterizing anthropogenic structures suggests using a sub-pixel mapping approach for analysing related LC from space. We implement a regression-based unmixing approach for mapping built-up surfaces and infrastructure, woody vegetation and non-woody vegetation for all of Germany and Austria at 10 m resolution to demonstrate the potential of sub-pixel mapping. We map LC fractions for one point in time, using all available Sentinel-2 data from 2017 and 2018 ( less then 70% cloud cover). We combine the concept of synthetically mixed training data with statistical aggregations from spectral-temporal metrics (STM) derived from Sentinel-2 reflectance time series. We specifically examine how STM can be used for creating synt8, 0.22) and non-woody vegetation (0.14, 0.19) are highly consistent across Germany and Austria. Only a few surface types were not accurately predicted in our nation-wide mapping. Further research is required to optimize mapping of temporally invariant bare soil and rock surfaces that show spectral similarity to built-up surfaces and infrastructure. The proposed methodology combines benefits of both regression-based modelling with synthetically mixed training data and STM, and thus facilitates mapping of LC fractions on a national scale and at high resolution. Such information will allow to better characterize settlements and identifying processes such as densification that are best represented by continuous LC mapping.Dithiolanes are used to obtain dynamic and reversible crosslinks between polymer chains. Copolymers of two different dithiolane-containing cyclic carbonate monomers and ε-caprolactone (CL) were synthesized by ring-opening polymerization using a methoxy-poly(ethylene glycol) (mPEG) initiator and different catalysts (diphenyl phosphate (DPP), methanesulfonic acid (MSA), 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD), or Sn(Oct)2). Each catalyst required a different temperature, which had a pronounced influence on the reactivity ratio of the monomers and occurrence of transesterification reactions and, therefore, the monomer sequence. Self-crosslinkable copolymers were obtained when the dithiolane units were connected closely to the polymer backbone, whereas the presence of a linker unit between the dithiolane and the backbone prevented self-crosslinking. The former amphiphilic PEGylated block copolymers formed micelles by nanoprecipitation in the aqueous environment and crosslinked spontaneously by disulfide exchange during subsequent dialysis. These dithiolane-crosslinked micelles showed reduction-responsive dissociation in the presence of 10 mM glutathione, making them promising drug delivery systems for the intracellularly triggered cargo release.The changes in the gluten network during extrusion treatment were studied by assessing the polymerization behavior of glutenin. Gluten samples were extruded at different barrel temperatures, screw speeds, and flow rates. The results indicated that high molecular weight glutenin subunits increased while free sulfhydryl groups and low molecular weight glutenin subunits decreased as the screw speeds and flow rates increased during extrusion treatment. Specific β-sheet structures of gluten clearly increased, while α-helices and β-turns fluctuated during extrusion processing, thus forming a tight gluten network. The characteristics of the protein network were evaluated by confocal laser scanning microscopy. The results showed that a homogeneous and denser gluten network was formed at higher extrusion temperatures during the extrusion process, which may be related to the polymerization of low-molecular-weight glutenin subunits. This study provides a theoretical basis for the improvement and regulation of extrusion quality during the gluten extrusion process.The demand for new gluten-free (GF) products is still very crucial issue in food industry. There is also a need for bioactive compounds and natural alternatives for food additives. For now, not only providing structure without gluten is major challenge, but also high sensory acceptance and nutritional value are on the top. This study is focused on the effect of high-purity oat β-glucan as a structure-making agent on physicochemical and sensory properties of gluten-free yeast leavened cake. The response surface methodology (RSM) was used to set the design of the experiment. Water and oat β-glucan were chosen as independent variables. Enzymatic extraction was conducted in order to obtain pure oat β-glucan (approx. 85%). Physicochemical and microstructure analyses, and a consumer hedonic test were carried out to check the quality of the final product. As a last step, verification was undertaken to compare the predicted and experimental values of the results. The results showed that the optimisation process was crucial in obtaining high-quality, gluten-free yeast leavened cake. The optimised amounts of water and oat β-glucan were 66.12% and 2.63% respectively. This proves that the application of oat β-glucan to gluten-free products is possible and gives positive results in terms of texture, volume and sensory acceptance. Due to oat β-glucan's pro-health benefits, the final product can be seen as a functional alternative for common gluten-free products in the market.Accurate calculation of electrostatic potential and gradient on the molecular surface is highly desirable for the continuum and hybrid modeling of large scale deformation of biomolecules in solvent. In this article a new numerical method is proposed to calculate these quantities on the dielectric interface from the numerical solutions of the Poisson-Boltzmann equation. Our method reconstructs a potential field locally in the least square sense on the polynomial basis enriched with Green's functions, the latter characterize the Coulomb potential induced by charges near the position of reconstruction. This enrichment resembles the decomposition of electrostatic potential into singular Coulomb component and the regular reaction field in the Generalized Born methods. Numerical experiments demonstrate that the enrichment recovery produces drastically more accurate and stable potential gradients on molecular surfaces compared to classical recovery techniques.In this work, a membraneless microbial fuel cell (MFC) with an empty volume of 1.5 mL, fed continuously with hydrolysed urine, was tested in supercapacitive mode (SC-MFC). In order to enhance the power output, a double strategy was used i) a double cathode was added leading to a decrease in the equivalent series resistance (ESR); ii) the apparent capacitance was boosted up by adding capacitive features on the anode electrode. Galvanostatic (GLV) discharges were performed at different discharge currents. The results showed that both strategies were successful obtaining a maximum power output of 1.59 ± 0.01 mW (1.06 ± 0.01 mW mL-1) at pulse time of 0.01 s and 0.57 ± 0.01 mW (0.38 ± 0.01 mW mL-1) at pulse time of 2 s. The highest energy delivered at ipulse equal to 2 mA was 3.3 ± 0.1 mJ. The best performing SC-MFCs were then connected in series and parallel and tested through GLV discharges. As the power output was similar, the connection in parallel allowed to roughly doubling the current produced. Durability tests over ≈5.6 days showed certain stability despite a light overall decrease.This work is presenting for the first time the use of inexpensive and efficient anode material for boosting power production, as well as improving electrofiltration of human urine in tubular microbial fuel cells (MFCs). The MFCs were constructed using unglazed ceramic clay functioning as the membrane and chassis. The study is looking into effective anodic surface modification by applying activated carbon micro-nanostructure onto carbon fibres that allows electrode packing without excessive enlargement of the electrode. The surface treatment of the carbon veil matrix resulted in 3.7 mW (52.9 W m-3 and 1626 mW m-2) of power generated and almost a 10-fold increase in the anodic current due to the doping as well as long-term stability in one year of continuous operation. The higher power output resulted in the synthesis of clear catholyte, thereby i) avoiding cathode fouling and contributing to the active splitting of both pH and ions and ii) transforming urine into a purified catholyte - 30% salt reduction - by electroosmotic drag, whilst generating - rather than consuming - electricity, and in a way demonstrating electrofiltration. For the purpose of future technology implementation , the importance of simultaneous increase in power generation, long-term stability over 1 year and efficient urine cleaning by using low-cost materials, is very promising and helps the technology enter the wider market.Coronavirus disease 2019 (COVID-19) in humans is often a clinically mild illness, but some individuals develop severe pneumonia, respiratory failure and death1-4. Studies of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in hamsters5-7 and nonhuman primates8-10 have generally reported mild clinical disease, and preclinical SARS-CoV-2 vaccine studies have demonstrated reduction of viral replication in the upper and lower respiratory tracts in nonhuman primates11-13. Here we show that high-dose intranasal SARS-CoV-2 infection in hamsters results in severe clinical disease, including high levels of virus replication in tissues, extensive pneumonia, weight loss and mortality in a subset of animals. A single immunization with an adenovirus serotype 26 vector-based vaccine expressing a stabilized SARS-CoV-2 spike protein elicited binding and neutralizing antibody responses and protected against SARS-CoV-2-induced weight loss, pneumonia and mortality. These data demonstrate vaccine protection against SARS-CoV-2 clinical disease. This model should prove useful for preclinical studies of SARS-CoV-2 vaccines, therapeutics and pathogenesis.The process of poly(ADP-ribosyl)ation and the major enzyme that catalyses this reaction, poly(ADP-ribose) polymerase 1 (PARP1), were discovered more than 50 years ago. Since then, advances in our understanding of the roles of PARP1 in cellular processes such as DNA repair, gene transcription and cell death have allowed the investigation of therapeutic PARP inhibition for a variety of diseases - particularly cancers in which defects in DNA repair pathways make tumour cells highly sensitive to the inhibition of PARP activity. Efforts to identify and evaluate potent PARP inhibitors have so far led to the regulatory approval of four PARP inhibitors for the treatment of several types of cancer, and PARP inhibitors have also shown therapeutic potential in treating non-oncological diseases. This Review provides a timeline of PARP biology and medicinal chemistry, summarizes the pathophysiological processes in which PARP plays a role and highlights key opportunities and challenges in the field, such as counteracting PARP inhibitor resistance during cancer therapy and repurposing PARP inhibitors for the treatment of non-oncological diseases.Sandhoff disease (SD) is an autosomal recessive lysosomal storage disease caused by defects in the β-subunit of β-N-acetylhexosaminidase (Hex), the enzyme that catabolizes GM2 ganglioside. Hex deficiency causes neuronal storage of GM2 and related glycoconjugates, resulting in progressive neurodegeneration and death, typically in infancy. No effective treatment exists for human patients. Adeno-associated virus (AAV) gene therapy led to improved clinical outcome and survival of SD cats treated before the onset of disease symptoms. Most human patients are diagnosed after clinical disease onset, so it is imperative to test AAV-gene therapy in symptomatic SD cats to provide a realistic indication of therapeutic benefits that can be expected in humans. In this study, AAVrh8 vectors injected into the thalamus and deep cerebellar nuclei of symptomatic SD cats resulted in widespread central nervous system enzyme distribution, although a substantial burden of storage material remained. Cats treated in the early symptomatic phase showed delayed disease progression and a significant survival increase versus untreated cats. Treatment was less effective when administered later in the disease course, although therapeutic benefit was still possible. Results are encouraging for the treatment of human patients and provide support for the development AAV-gene therapy for human SD.An amendment to this paper has been published and can be accessed via a link at the top of the paper.Intestinal stem and progenitor cells replicate and differentiate in distinct compartments, influenced by Wnt, BMP, and other subepithelial cues. The cellular sources of these signals were long obscure because intestinal mesenchyme was insufficiently characterised. In this Review, we discuss how recent mRNA profiles of mouse and human intestinal submucosa, coupled with fine-resolution microscopy and gene and cell disruptions, reveal a coherent picture of an organised tissue carrying cells with distinct molecular properties and functions.Transient modulation of the genes involved in immunity, without exerting a permanent change in the DNA code, can be an effective strategy to modulate the course of many inflammatory conditions. CRISPR-Cas9 technology represents a promising platform for achieving this goal. Truncation of guide RNA (gRNA) from the 5' end enables the application of a nuclease competent Cas9 protein for transcriptional modulation of genes, allowing multifunctionality of CRISPR. Here, we introduce an enhanced CRISPR-based transcriptional repressor to reprogram immune homeostasis in vivo. In this repressor system, two transcriptional repressors-heterochromatin protein 1 (HP1a) and Krüppel-associated box (KRAB)-are fused to the MS2 coat protein and subsequently recruited by gRNA aptamer binding to a nuclease competent CRISPR complex containing truncated gRNAs. With the enhanced repressor, we demonstrate transcriptional repression of the Myeloid differentiation primary response 88 (Myd88) gene in vitro and in vivo. We demonstrate thawe report that CRISPR-mediated repression of endogenous Myd88 can effectively modulate the host immune response against AAV-mediated gene therapy and influence the course of septicaemia. The ability to control Myd88 transcript levels using a CRISPR-based synthetic repressor can be an effective strategy for AAV-based CRISPR therapies, as this pathway serves as a key node in the induction of humoral immunity against AAV serotypes.Adaptive behaviour crucially depends on flexible decision-making, which in mammals relies on the frontal cortex, specifically the orbitofrontal cortex (OFC)1-9. How OFC encodes decision variables and instructs sensory areas to guide adaptive behaviour are key open questions. Here we developed a reversal learning task for head-fixed mice, monitored the activity of neurons of the lateral OFC using two-photon calcium imaging and investigated how OFC dynamically interacts with primary somatosensory cortex (S1). Mice learned to discriminate 'go' from 'no-go' tactile stimuli10,11 and adapt their behaviour upon reversal of stimulus-reward contingency ('rule switch'). Imaging individual neurons longitudinally across all behavioural phases revealed a distinct engagement of S1 and lateral OFC, with S1 neural activity reflecting initial task learning, whereas lateral OFC neurons responded saliently and transiently to the rule switch. We identified direct long-range projections from lateral OFC to S1 that can feed this activity back to S1 as value prediction error. This top-down signal updated sensory representations in S1 by functionally remapping responses in a subpopulation of neurons that was sensitive to reward history. Functional remapping crucially depended on top-down feedback as chemogenetic silencing of lateral OFC neurons disrupted reversal learning, as well as plasticity in S1. The dynamic interaction of lateral OFC with sensory cortex thus implements computations critical for value prediction that are history dependent and error based, providing plasticity essential for flexible decision-making.An amendment to this paper has been published and can be accessed via a link at the top of the paper.The voltage-gated potassium channel KCNQ2 is responsible for M-current in neurons and is an important drug target to treat epilepsy, pain and several other diseases related to neuronal hyper-excitability. A list of synthetic compounds have been developed to directly activate KCNQ2, yet our knowledge of their activation mechanism is limited, due to lack of high-resolution structures. Here, we report cryo-electron microscopy (cryo-EM) structures of the human KCNQ2 determined in apo state and in complex with two activators, ztz240 or retigabine, which activate KCNQ2 through different mechanisms. The activator-bound structures, along with electrophysiology analysis, reveal that ztz240 binds at the voltage-sensing domain and directly stabilizes it at the activated state, whereas retigabine binds at the pore domain and activates the channel by an allosteric modulation. By accurately defining ligand-binding sites, these KCNQ2 structures not only reveal different ligand recognition and activation mechanisms, but also provide a structural basis for drug optimization and design.In Gram-negative bacteria, phospholipids are major components of the inner membrane and the inner leaflet of the outer membrane, playing an essential role in forming the unique dual-membrane barrier to exclude the entry of most antibiotics. Understanding the mechanisms of phospholipid translocation between the inner and outer membrane represents one of the major challenges surrounding bacterial phospholipid homeostasis. The conserved MlaFEDB complex in the inner membrane functions as an ABC transporter to drive the translocation of phospholipids between the inner membrane and the periplasmic protein MlaC. However, the mechanism of phospholipid translocation remains elusive. Here we determined three cryo-EM structures of MlaFEDB from Escherichia coli in its nucleotide-free and ATP-bound conformations, and performed extensive functional studies to verify and extend our findings from structural analyses. Our work reveals unique structural features of the entire MlaFEDB complex, six well-resolved phospholipids in three distinct cavities, and large-scale conformational changes upon ATP binding. Together, these findings define the cycle of structural rearrangement of MlaFEDB in action, and suggest that MlaFEDB uses an extrusion mechanism to extract and release phospholipids through the central translocation cavity.Proper development of fetal germ cells (FGCs) is vital for the precise transmission of genetic and epigenetic information through generations. The transcriptional landscapes of human FGC development have been revealed; however, the epigenetic reprogramming process of FGCs remains elusive. Here, we profiled the genome-wide DNA methylation and chromatin accessibility of human FGCs at different phases as well as gonadal niche cells at single-cell resolution. First, we found that DNA methylation levels of FGCs changed in a temporal manner, whereas FGCs at different phases in the same embryo exhibited comparable DNA methylation levels and patterns. Second, we revealed the phase-specific chromatin accessibility signatures at the promoter regions of a large set of critical transcription factors and signaling pathway genes. We also identified potential distal regulatory elements including enhancers in FGCs. Third, compared with other hominid-specific retrotransposons, SVA_D might have a broad spectrum of binding capacity for transcription factors, including SOX15 and SOX17. Finally, using an in vitro culture system of human FGCs, we showed that the BMP signaling pathway promoted the cell proliferation of FGCs, and regulated the WNT signaling pathway by orchestrating the chromatin accessibility of its ligand genes. Our single-cell epigenomic atlas and functional assays provide valuable insights for understanding the strongly heterogeneous, unsynchronized, yet highly robust nature of human germ cell development.An amendment to this paper has been published and can be accessed via a link at the top of the paper.

To obtain a picture of the current status, training and governance for advanced practice and extended roles in the ophthalmic hospital non-medical workforce.

A 10 question, quantitative survey was designed with multidisciplinary members of the UK Ophthalmology Alliance and sent to the membership to obtain information on expanded non-medical roles.

34 of the 58 UKOA member hospitals responded (58% response rate). All responding units were using registered optometrists, orthoptists and nurses to undertake expanded outpatient roles and 28/34 (82%) had expanded roles for undertaking procedures. Some units had large numbers of staff undertaking these roles. There were noticeable trends for certain professional groups to undertake certain roles. For example, nurses were undertaking most procedures, apart from lasers which were mainly delivered by optometrists. Nurses had the lowest banding and optometrists the highest for apparently similar roles. Training was mostly in-house apprenticeship style although some formal external qualifications were undertaken.

Ophthalmology is developing many innovative roles for the non-medical workforce and, with the launch of the OCCCF training, this is likely to increase. Terminology is confusing and a categorisation suitable for ophthalmology is proposed.

Ophthalmology is developing many innovative roles for the non-medical workforce and, with the launch of the OCCCF training, this is likely to increase. Terminology is confusing and a categorisation suitable for ophthalmology is proposed.

Vascular malformations (VM) are primarily caused by somatic activating pathogenic variants in oncogenes. Targeted pharmacotherapies are emerging but require molecular diagnosis. Since variants are currently only detected in malformation tissue, patients may be ineligible for clinical trials prior to surgery. We hypothesized that cell-free DNA (cfDNA) could provide molecular diagnoses for patients with isolated VM.

cfDNA was isolated from plasma or cyst fluid from patients with arteriovenous malformations (AVM), venous malformations (VeM), or lymphatic malformations (LM), and assayed for known pathogenic variants using droplet digital polymerase chain reaction (ddPCR). Cyst fluid cfDNA from an independent cohort of LM patients was prospectively screened for variants using a multiplex ddPCR assay.

Variants were detected in plasma cfDNA in patients with AVM (2/8) and VeM (1/3). Variants were detected in cyst fluid cfDNA (7/7) but not plasma (0/26) in LM patients. Prospective testing of cyst fluid cfDNA with multiplex ddPCR identified variants in LM patients who had never undergone surgery (4/5).

Variants were detected in plasma from AVM and VeM patients, and in cyst fluid from patients with LM. These data support investigation of cfDNA-based molecular diagnostics for VM patients, which may provide opportunities to initiate targeted pharmacotherapies without prior surgery.

Variants were detected in plasma from AVM and VeM patients, and in cyst fluid from patients with LM. These data support investigation of cfDNA-based molecular diagnostics for VM patients, which may provide opportunities to initiate targeted pharmacotherapies without prior surgery.

To achieve the ultimate goal of personalized treatment of patients, accurate molecular diagnosis and precise interpretation of the impact of genetic variants on gene function is essential. With sequencing cost becoming increasingly affordable, the accurate distinguishing of benign from pathogenic variants becomes the major bottleneck. Although large normal population sequence databases have become a key resource in filtering benign variants, they are not effective at filtering extremely rare variants.

To address this challenge, we developed a novel statistical test by combining sequencing data from a patient cohort with a normal control population database. By comparing the expected and observed allele frequency in the patient cohort, variants that are likely benign can be identified.

The performance of this new method is evaluated on both simulated and real data sets coupled with experimental validation. As a result, we demonstrate this new test is well powered to identify benign variants, and is particularly effective for variants with low frequency in the normal population.

Overall, as a general test that can be applied to any type of variants in the context of all Mendelian diseases, our work provides a general framework for filtering benign variants with very low population allele frequency.

Overall, as a general test that can be applied to any type of variants in the context of all Mendelian diseases, our work provides a general framework for filtering benign variants with very low population allele frequency.An amendment to this paper has been published and can be accessed via a link at the top of the paper.Phosphatase and tensin homolog (PTEN) loss is associated with adverse outcomes in prostate cancer and has clinical potential as a prognostic biomarker. The objective of this work was to develop an artificial intelligence (AI) system for automated detection and localization of PTEN loss on immunohistochemically (IHC) stained sections. PTEN loss was assessed using IHC in two prostate tissue microarrays (TMA) (internal cohort, n = 272 and external cohort, n = 129 patients). TMA cores were visually scored for PTEN loss by pathologists and, if present, spatially annotated. Cores from each patient within the internal TMA cohort were split into 90% cross-validation (N = 2048) and 10% hold-out testing (N = 224) sets. ResNet-101 architecture was used to train core-based classification using a multi-resolution ensemble approach (×5, ×10, and ×20). For spatial annotations, single resolution pixel-based classification was trained from patches extracted at ×20 resolution, interpolated to ×40 resolution, and applied in a sples. AI-based algorithms have potential to streamline sample assessment in research and clinical laboratories.PD-L1 immunohistochemistry (IHC) currently has the most Food and Drug Administration (FDA) approvals as a companion diagnostic (CDx) for immunotherapies in specific tumor types; however, multiple other immunotherapy biomarkers exist. We performed this study to examine and report the prevalence of PD-L1 expression in a wide variety of tumor types and examine its relationship to microsatellite instability (MSI), tumor mutational burden (TMB), and CD274 (PD-L1) gene amplification. We performed a retrospective analysis of all cases in which both PD-L1 IHC (using the DAKO 22C3 IHC assay with either tumor proportion score (TPS) or combined positive score (CPS); or the VENTANA SP142 assay with infiltrating immune cell score (IC)) and comprehensive genomic profiling (CGP) were tested at Foundation Medicine between January 2016 and November 2019. Of note, PD-L1 positivity is defined per the CDx indication and tumor proportion score (TPS ≥ 1) for indications without a CDx claim; and TMB positivity is defined as ≥10 mutpy.Immune checkpoint inhibitors (ICI) can induce a durable response against a wide range of malignancies but cause immune related adverse events. The purpose of this study was to evaluate whether the pattern of inflammation in a liver biopsy in patients on ICIs is likely to be related to ICIs or other causes, and whether the pattern correlates with LFT abnormalities, imaging findings, and responsiveness to steroids. Cancer patients on ICIs who underwent liver biopsy were identified. Clinical data were obtained from electronic records. Liver biopsies were recorded as hepatitic, cholangitic, mixed, steatotic, or as mild nonspecific changes. In total, 28 liver biopsies had a predominantly hepatitic pattern of inflammation, including 11 biopsies with granulomas and 10 with endothelialitis. Eight biopsies had a mixed hepatocytic and cholangitic pattern of injury, including 6 with granulomas and 4 with endothelialitis. Sixteen patients had a predominantly cholangitic pattern, with portal-based inflammation. Three patients had a pattern resembling fatty liver, and five had mild nonspecific changes. The three most common histologic patterns correlated with the pattern of LFT abnormalities. The majority of patients with a cholangitic pattern had competing causes for elevated LFTs, including disease progression or concomitant chemotherapy. The cholangitic pattern was more likely to have bile duct dilatation or narrowing on liver imaging. The pattern of inflammation, degree of lobular injury, or presence of granulomas or endothelialitis did not predict response to steroids or the need for secondary immunosuppression. In this retrospective study, the pattern of inflammation did not predict the need for steroids, the length of time that steroids is required, or the need for secondary immunosuppression. A cholangitic pattern was seen when the pattern of LFTs was cholestatic, and was associated with imaging abnormalities of the bile duct, but a similar pattern was seen in bile duct obstruction and other drug reactions.An amendment to this paper has been published and can be accessed via a link at the top of the paper.Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by antinuclear antibodies (ANAs) that form immune complexes that mediate pathogenesis by tissue deposition or cytokine induction. Some ANAs bind DNA or associated nucleosome proteins, whereas other ANAs bind protein components of complexes of RNA and RNA-binding proteins (RBPs). Levels of anti-DNA antibodies can fluctuate widely, unlike those of anti-RBP antibodies, which tend to be stable. Because anti-DNA antibody levels can reflect disease activity, repeat testing is common; by contrast, a single anti-RBP antibody determination is thought to suffice for clinical purposes. Experience from clinical trials of novel therapies has provided a new perspective on ANA expression during disease, as many patients with SLE are ANA negative at screening despite previously testing positive. Because trial results suggest that patients who are ANA negative might not respond to certain agents, screening strategies now involve ANA and anti-DNA antibody testing to identify patients with so-called 'active, autoantibody-positive SLE'. Evidence suggests that ANA responses can decrease over time because of the natural history of disease or the effects of therapy. Together, these findings suggest that, during established disease, more regular serological testing could illuminate changes relevant to pathogenesis and disease status.Traditional Chinese medicine (TCM) is an entirely coherent system, with internal logic and consistency of thought and practice. Though TCM has a long history, it is not easily accepted by Western medicine due to its theoretical and conceptual complexity. TCM nutrition is an ancient but burgeoning discipline, and its main goal is to use food as a means to achieve balance and harmony within the body. Compared with modern nutrition, it has unique beneficial concepts, such as the holism, diet suggestions based on syndrome differentiation, the idea that the spleen-stomach is the "root" of post-heaven, and the homology of medicine and food. Until today, it is difficult to evaluate whether TCM nutrition could play a major role in the treatment of various diseases. The limitations mainly include the scope of application is limited, lack of evidence-based research, and the constitution differentiation need the cooperation of clinicians of TCM. In contemporary China, the inheritance, innovation, and broadening the scope of applications of TCM nutrition is very important. The government should establish a system in which TCM nutrition and modern nutrition coexist, and perform higher specialist training for dietitians of TCM. Moreover, TCM nutrition should integrate the research methods of modern nutrition, and involve adjustment to target populations, the formulation of age-specific nutrition principles, and an emphasis on the research and development of nutritional food, thus fully demonstrating the advantages and characteristics of TCM nutrition.Aluminum (Al) contamination of parenteral nutrition (PN) solutions has been known for over 30 years. In particular, vascular intake of Al leads to its accumulation in tissues. In this study, 8 all-in-one PN solutions the aluminum concentration was analyzed by high-performance liquid chromatography. The mean Al concentration of the glucose solutions of the PN solutions combinations was 16.36 ± 8.31 µg/L, the mean Al concentration of the amino acid solutions was 4.96 ± 3.73 µg/L, and the mean Al concentration of the lipid solutions was 9.09 ± 11.23 µg/L. The Al concentration of the PN5 glucose and PN2 lipid solutions were above 25 µg/L, which is the limit set by the Food and Drug Administration (FDA). No studies in the literature have examined the Al concentrations of all-in-one PN solutions via HPLC. In two of the analyzed solutions, the Al concentration was found to be higher than the limit set by the FDA.The 'real-world' patient population of metastatic melanoma is not fully represented in clinical trials investigating checkpoint inhibitors. We described therapy discontinuation in an unselected population-based cohort of adults with metastatic melanoma who started therapy with pembrolizumab, nivolumab, or nivolumab/ipilimumab from January 2015 to August 2017. Therapy discontinuation was defined as a gap between doses beyond 120 days, and/or initiation of another cancer therapy. We estimated drug-specific rate ratios for therapy discontinuation adjusted for age, sex, comorbidities, health care use, and past cancer therapies. We included 876 metastatic melanoma patients initiating pembrolizumab (44.3%), nivolumab/ipilimumab (31.2%), and nivolumab (24.5%). At 12 months of follow-up, the probabilities of therapy discontinuation were 49.9% (95% confidence interval, CI 43.6-56.5) for pembrolizumab, 58.8% (95% CI 50.5-67.3) for nivolumab, and 59.2% (95% CI 51.7-66.8) for nivolumab/ipilimumab. Stratified analyses based on prior cancer therapy, brain metastases at baseline, and sex showed similar trends. In multivariable analyses, compared with pembrolizumab, patients starting nivolumab (rate ratio 1.38, 95% CI 1.08-1.77) or nivolumab/ipilimumab (rate ratio 1.30, 95% CI 1.02-1.65) were more likely to discontinue therapy. Our findings indicate frequent discontinuations of checkpoint inhibitors at one year. The lower discontinuation associated with pembrolizumab should be confirmed in further studies.In this study, we performed a spinal muscular atrophy carrier screening investigation with NGS-based method. First, the validation for NGS-based method was implemented in 2255 samples using real-time PCR. The concordance between the NGS-based method and real-time PCR for the detection of SMA carrier and patient were up to 100%. Then, we applied this NGS-based method in 10,585 self-reported normal couples (34 Chinese ethnic groups from 5 provinces in South China) for SMA carrier screening. The overall carrier frequency was 1 in 73.8 (1.4%). It varied substantially between ethnic groups, highest in Dai ethnicity (4.3%), and no significant difference was found between five provinces. One couple was detected as carriers with an elevated risk of having an SMA affected baby. The distribution of SMN1SMN2 genotype was also revealed in this study. Among the individuals with normal phenotype, the exon 7 copy-number ratio of SMN1 to SMN2 proved the gene conversion between them. With NGS-based method, we investigated SMA carrier status in Chinese population for the first time, and our results demonstrated that it is a promising alternative for SMA carrier screening and could provide data support and reference for future clinical application.