Single cell sequencing data identify distinct B cell and fibroblast populations in stricturing Crohn's disease.

Single cell RNA sequencing of human full thickness Crohn's disease (CD) small bowel resection specimens was used to identify potential therapeutic targets for stricturing (S) CD. Using an unbiased approach, 16 cell lineages were assigned within 14,539 sequenced cells from patient-matched SCD and non-stricturing (NSCD) preparations. SCD and NSCD contained identical cell types. Amongst immune cells, B cells and plasma cells were selectively increased in SCD samples. B cell subsets suggested formation of tertiary lymphoid tissue in SCD and compared with NSCD there was an increase in IgG, and a decrease in IgA plasma cells, consistent with their potential role in CD fibrosis. Two Lumican-positive fibroblast subtypes were identified and subclassified based on expression of selectively enriched genes as fibroblast clusters (C) 12 and C9. Cells within these clusters expressed the profibrotic genes Decorin (C12) and JUN (C9). C9 cells expressed ACTA2; ECM genes COL4A1, COL4A2, COL15A1, COL6A3, COL18A1 and ADAMDEC1; LAMB1 and GREM1. GO and KEGG Biological terms showed extracellular matrix and stricture organization associated with C12 and C9, and regulation of WNT pathway genes with C9. Trajectory and differential gene analysis of C12 and C9 identified four sub-clusters. Intra sub-cluster gene analysis detected 13 co-regulated gene modules that aligned along predicted pseudotime trajectories. CXCL14 and ADAMDEC1 were key markers in module 1. Our findings support further investigation of fibroblast heterogeneity and interactions with local and circulating immune cells at earlier time points in fibrosis progression. Breaking these interactions by targeting one or other population may improve therapeutic management for SCD.

Human Intestinal Dendritic Cells Can Overcome Retinoic Acid Signaling to Generate Proinflammatory CD4 T Cells with Both Gut and Skin Homing Properties.

Retinoic acid, produced by intestinal dendritic cells (DCs), promotes T cell trafficking to the intestinal mucosa by upregulating α4ß7 integrin and inhibiting the generation of cutaneous leukocyte Ag (CLA) required for skin entry. In the present study, we report that activation of human naive CD4 T cells in an APC-free system generates cells expressing α4ß7 alone; in contrast, activation by intestinal DCs that produce retinoic acid and induce high levels of α4ß7 also results in CLA expression, generating CLA+α4ß7+ "dual tropic" cells, with both gut and skin trafficking potential, that also express high levels of α4ß1 integrin. DC generation of CLA+α4ß7+ T cells is associated with upregulation of FUT7, a fucosyltransferase involved in CLA generation; requires cell contact; and is enhanced by IL-12/IL-23. The blood CD4+ T cell population contains CLA+α4ß7+ cells, which are significantly enriched for cells capable of IFN-γ, IL-17, and TNF-α production compared with conventional CLA-α4ß7+ cells. Dual tropic lymphocytes are increased in intestinal tissue from patients with Crohn's disease, and single-cell RNA-sequencing analysis identifies a transcriptionally distinct cluster of FUT7-expressing cells present only in inflamed tissue; expression of genes associated with cell proliferation suggests that these cells are undergoing local activation. The expression of multiple trafficking molecules by CLA+α4ß7+ T cells can enable their recruitment by alternative pathways to both skin and gut; they may contribute to both intestinal and cutaneous manifestations of inflammatory bowel disease.

The Biomechanics Research and Innovation Challenge: Development, Implementation, Uptake, and Reflections on the Inaugural Program.

Biomechanics as a discipline is ideally placed to increase awareness and participation of girls and women in science, technology, engineering, and mathematics. A nationwide Biomechanics and Research Innovation Challenge (BRInC) centered on mentoring and role modeling was developed to engage high school girls (mentees) and early-mid-career women (mentors) in the field of biomechanics through the completion of a 100-day research and/or innovation project. This manuscript describes the development, implementation, and uptake of the inaugural BRInC program and synthesizes the research and innovation projects undertaken, providing a framework for adoption of this program within the global biomechanics community. Eighty-seven high school girls in years 9 and 10 (age range: 14-16 y) were mentored in teams (n = 17) by women in biomechanics (n = 24). Using a design thinking approach, teams generated solutions to biomechanics-based problem(s)/research question(s). Eight key reflections on program strengths, as well as areas for improvement and planned changes for future iterations of the BRInC program, are outlined. These key reflections highlight the innovation, impact, and scalability of the program; the importance of a program framework and effective communication tools; and implementation of strategies to sustain the program as well as the importance of diversity and building a sense of community.

A Dual Inertial Measurement Unit System for Classifying Standard Overhead Drill Movements in Elite Women's Water Polo.

INTRODUCTION: Water polo upper limb external load monitoring cannot be currently measured accurately because of technological and methodological challenges. This is problematic as large fluctuations in overhead movement volume and intensity may affect performance and alter injury risk. Inertial measurement units (IMU) and machine learning techniques have been shown to accurately classify overhead movements in other sports. We investigated the model accuracy and class precision, sensitivity, and specificity of IMU and machine learning techniques to classify standard overhead drill movements in elite women's water polo. METHODS: Ten women's water polo players performed standard drills of swimming, blocking, low-intensity throwing and high-intensity throwing under training conditions. Athletes wore two IMU: one on the upper back and the other on the distal forearm. Each movement was videoed and coded to a standard overhead drill movement. IMU and coded video data were merged to verify the IMU-detected activity classification of each movement to that of the video. Data were partitioned into a training and a test set and used to form a decision tree algorithm. Model accuracy and class precision, sensitivity, and specificity were assessed. RESULTS: IMU resultant acceleration and angular velocity values displayed drill-specific values. A total of 194 activities were identified by the model in the test set, with 8 activities being incorrectly classified. Model accuracy was 95.88%. Percentage class precision, sensitivity, and specificity were as follows: blocking (96.15, 86.21, 99.39), high-intensity throwing (100, 100, 100), low-intensity throwing (93.48, 93.48, 97.97), and swimming (94.81, 98.65, 96.67). CONCLUSIONS: IMU and machine learning techniques can accurately classify standard overhead drill movements in elite women's water polo.

Epigenetic and metabolic reprogramming of fibroblasts in Crohn's disease strictures reveals histone deacetylases as therapeutic targets.

BACKGROUND & AIMS: No effective therapeutic intervention exists for intestinal fibrosis in Crohn's disease [CD]. We characterised fibroblast subtypes, epigenetic and metabolic changes, and signalling pathways in CD fibrosis to inform future therapeutic strategies. METHODS: We undertook immunohistochemistry, metabolic, signalling pathway and Epigenetic [Transposase-Accessible Chromatin using sequencing] analyses associated with collagen production in CCD-18Co intestinal fibroblasts and primary fibroblasts isolated from stricturing [SCD] and non-stricturing [NSCD] CD small intestine. SCD/ NSCD fibroblasts were cultured with TGFß and valproic acid [VPA]. RESULTS: Stricturing CD was characterised by distinct histone deacetylase [HDAC] expression profiles, particularly HDAC1, HDAC2, and HDAC7. As a proxy for HDAC activity, reduced numbers of H3K27ac+ cells were found in SCD compared to NSCD sections. Primary fibroblasts had increased extracellular lactate [increased glycolytic activity] and intracellular hydroxyproline [increased collagen production] in SCD compared to NSCD cultures. The metabolic effect of TGFß-stimulation was reversed by the HDAC inhibitor VPA. SCD fibroblasts appear "metabolically primed" and responded more strongly to both TGFß and VPA. Treatment with VPA revealed TGFß-dependent and independent Collagen-I production in CCD-18Co cells and primary fibroblasts. VPA altered the epigenetic landscape with reduced chromatin accessibility at the COL1A1 and COL1A2 promoters. CONCLUSIONS: Increased HDAC expression profiles, H3K27ac hypoacetylation, a significant glycolytic phenotype, and metabolic priming, characterise SCD-derived as compared to NSCD fibroblasts. Our results reveal a novel epigenetic component to Collagen-I regulation and TGFß-mediated CD fibrosis. HDAC inhibitor therapy may 'reset' the epigenetic changes associated with fibrosis.

Macrocystis pyrifera ferment-containing creams for optimizing facial skin rejuvenation.

BACKGROUND: There is an increasing demand for facial skin rejuvenation. Specialized aesthetic skincare treatments may be one of the first steps to help prevent or treat facial signs of aging. This article discusses aesthetic skin care for facial skin rejuvenation, particularly data on two creams containing Macrocystis pyrifera ferment. METHODS: The authors convened a dermatology advisory board to discuss challenges and practices in using skincare for facial rejuvenation, combining their expert opinion and experience on facial rejuvenation with preclinical and clinical data on two creams containing Macrocystis pyrifera ferment and a review of the literature. RESULTS: Preclinical and clinical studies on Macrocystis pyrifera ferment and two creams containing the ferment exhibit anti-inflammatory, anti-aging, and healing properties. In preclinical studies, the ferment demonstrated collagen type I enhancing properties in ex vivo skin models, and skin cells treated with the ferment migrated faster than untreated cells in the in vitro study. In clinical studies measuring visible anti-inflammatory activity, the ferment alone and the ferment-containing products significantly decreased erythema, and in anti-aging studies, they improved visible skin aging parameters. Finally, in clinical studies on the stratum corneum, the two creams increased moisture levels and decreased transepidermal water loss (TEWL), reflecting healing by enhancing barrier strength and recovery. CONCLUSIONS: The Macrocystis pyrifera ferment and creams containing the ferment are effective skin care treatment products to decrease the visible effects of inflammation and signs of aging while promoting healing by enhancing barrier resilience and recovery.

Treatment of mild to severe acne with 1726 nm laser: A safe alternative to traditional acne therapies.

INTRODUCTION: Acne is the most common reason for dermatology consultation in adolescents and young adults. Consultation is often delayed despite unsuccessful self-treatment. Postponing effective treatment places acne sufferers at higher risk for permanent acne scars and post-inflammatory pigment changes. AIM: This review discusses clinical challenges with present therapeutic options for acne treatment and the role of a 1726 nm laser for acne. METHODS: Current acne treatment guidelines were reviewed. A literature review was conducted for trials of light-based acne therapy. The selectivity of previous light-based therapies was reviewed. RESULTS: Available acne therapy is effective, but treatment-related side effects are common. Acne treatment guidelines do not include recommendations for light-based treatments. Different types of light-based treatments have been tried but until now no wavelength specifically targeted sebaceous glands. CONCLUSION: The 1726 nm laser is safe and effective for treating mild to severe acne in all Fitzpatrick skin types. Acne resolution is apparent within the first month and improves for up to 2 years beyond treatment.

Model ensembles of ecosystem services fill global certainty and capacity gaps.

Sustaining ecosystem services (ES) critical to human well-being is hindered by many practitioners lacking access to ES models ("the capacity gap") or knowledge of the accuracy of available models ("the certainty gap"), especially in the world's poorer regions. We developed ensembles of multiple models at an unprecedented global scale for five ES of high policy relevance. Ensembles were 2 to 14% more accurate than individual models. Ensemble accuracy was not correlated with proxies for research capacity, indicating that accuracy is distributed equitably across the globe and that countries less able to research ES suffer no accuracy penalty. By making these ES ensembles and associated accuracy estimates freely available, we provide globally consistent ES information that can support policy and decision-making in regions with low data availability or low capacity for implementing complex ES models. Thus, we hope to reduce the capacity and certainty gaps impeding local- to global-scale movement toward ES sustainability.

An arginase 2 promoter transgenic line illuminates immune cell polarisation in zebrafish.

Innate immune responses to inflammation and infection are complex and represent major challenges for developing much needed new treatments for chronic inflammatory diseases and drug-resistant infections. To be ultimately successful, the immune response must be balanced to allow pathogen clearance without excess tissue damage, processes controlled by pro- and anti-inflammatory signals. The roles of anti-inflammatory signalling in raising an appropriate immune response are underappreciated, representing overlooked potential drug targets. This is especially true in neutrophils, a difficult cell type to study ex vivo owing to a short lifespan, dogmatically seen as being highly pro-inflammatory. Here, we have generated and describe the first zebrafish transgenic line [TgBAC(arg2:eGFP)sh571] that labels expression of the anti-inflammatory gene arginase 2 (arg2) and show that a subpopulation of neutrophils upregulate arginase soon after immune challenge with injury and infection. At wound-healing stages, arg2:GFP is expressed in subsets of neutrophils and macrophages, potentially representing anti-inflammatory, polarised immune cell populations. Our findings identify nuanced responses to immune challenge in vivo, responses that represent new opportunities for therapeutic interventions during inflammation and infection.

Small-molecule Wnt inhibitors are a potential novel therapy for intestinal fibrosis in Crohns disease.

Intestinal fibrosis and stricture formation is an aggressive complication of Crohns disease (CD), linked to increased morbidity and costs. The present study investigates the contribution of Wingless-Int-1 (Wnt) signalling to intestinal fibrogenesis, considers potential cross-talk between Wnt and transforming growth factor ß1 (TGFß) signalling pathways, and assesses the therapeutic potential of small-molecule Wnt inhibitors. ß-catenin expression was explored by immunohistochemistry (IHC) in formalin-fixed paraffin embedded (FFPE) tissue from patient-matched nonstrictured (NSCD) and strictured (SCD) intestine (n=6 pairs). Functional interactions between Wnt activation, TGFß signalling, and type I collagen (Collagen-I) expression were explored in CCD-18Co cells and primary CD myofibroblast cultures established from surgical resection specimens (n=16) using small-molecule Wnt inhibitors and molecular techniques, including siRNA-mediated gene knockdown, immunofluorescence (IF), Wnt gene expression arrays, and western blotting. Fibrotic SCD tissue was marked by an increase in ß-catenin-positive cells. In vitro, activation of Wnt-ß-catenin signalling increased Collagen-I expression in CCD-18Co cells. Conversely, ICG-001, an inhibitor of ß-catenin signalling, reduced Collagen-I expression in cell lines and primary CD myofibroblasts. TGFß increased ß-catenin protein levels but did not activate canonical Wnt signalling. Rather, TGFß up-regulated WNT5B, a noncanonical Wnt ligand, and the Wnt receptor FZD8, which contributed directly to the up-regulation of Collagen-I through a ß-catenin-independent mechanism. Treatment of CCD-18Co fibroblasts and patient-derived myofibroblasts with the FZD8 inhibitor 3235-0367 reduced extracellular matrix (ECM) expression. Our data highlight small-molecule Wnt inhibitors of both canonical and noncanonical Wnt signalling, as potential antifibrotic drugs to treat SCD intestinal fibrosis. They also highlight the importance of the cross-talk between Wnt and TGFß signalling pathways in CD intestinal fibrosis.

Preference Testing in Medical Devices: Current Framework and Regulatory Gaps.

Preference testing is a valuable source of information that can be provided by both healthcare professionals (HCPs) and patients (users). It can be used to improve the design and development of medical devices by feeding into device usability and, ultimately, risk management. Furthermore, it can aid with selecting the most appropriate clinical endpoints to be used in the clinical evaluation of a device and increase patient engagement by incorporating patient-relevant outcomes. Preference testing is widely conducted in the food industry but is not widespread in the medical field due to limited guidelines and a lack of regulatory framework. As such, manufacturers may be unaware of the benefits of preference testing and fail to take full advantage of it, or conversely, may use inappropriate methodology and/or analyses and consequently fail to collect meaningful data. In this position paper, we aim to highlight the benefits and uses of preference testing, along with potential methods that could be used for preference testing of medical devices. A key step towards the wider implementation of preference testing in medical devices is for the publication of international standards and guidelines for the collection, assessment, and implementation of preference data into the life cycle of a medical device.

Throwing in the deep end: athletes, coaches and support staff experiences, perceptions and beliefs of upper limb injuries and training load in elite women's water polo.

To understand elite athlete, coach and support staff experiences, perceptions and beliefs in women's water polo with managing upper limb injuries and monitoring training loads. Inductive qualitative design. Twenty athletes, coaches and support staff were purposively recruited and participated in semistructured interviews. Participants either had experienced an upper limb injury or had experience managing athletes with upper limb injuries. Interviews were conducted in-person or virtually, audio-recorded, deidentified, transcribed verbatim and cleaned to ensure accuracy. Data were thematically analysed. Analysis identified five cohesive themes: (1) upper limb injury management is adequate-but prevention, communication and knowledge need improving, (2) current training load monitoring generates uncertainty and lack of consistency of processes-due to reliance on internal, and lack of external load monitoring, (3) optimal training load monitoring requires objective measurement of training load-that accurately measures the external load of athletes' upper limbs, (4) athlete-centred philosophy matters-including athlete-centred care to facilitate individually tailored rehabilitation programmes and their inclusion in management decisions, (5) mental, social and emotional aspects of upper limb injury management matter-acknowledging feelings of loss of team inclusion, fear of missing out and frustration felt by athletes as well as the emotional labour felt by coaches when supporting athletes with an upper limb injury. Upper limb injury management and training load monitoring are evolving areas where objective measurement of training load may assist in increasing consistency of communication, collaboration and coordination between all stakeholders, and to address uncertainty. Stakeholders placed value in intangible qualities such as trust and care in their relationships with other collaborators-facilitating athlete physical, mental and emotional recovery following upper limb injuries.

Companion Diagnostics: State of the Art and New Regulations.

Companion diagnostics (CDx) hail promise of improving the drug development process and precision medicine. However, there are various challenges involved in the clinical development and regulation of CDx, which are considered high-risk in vitro diagnostic medical devices given the role they play in therapeutic decision-making and the complications they may introduce with respect to their sensitivity and specificity. The European Union (E.U.) is currently in the process of bringing into effect in vitro Diagnostic Medical Devices Regulation (IVDR). The new Regulation is introducing a wide range of stringent requirements for scientific validity, analytical and clinical performance, as well as on post-market surveillance activities throughout the lifetime of in vitro diagnostics (IVD). Compliance with General Safety and Performance Requirements (GSPRs) adopts a risk-based approach, which is also the case for the new classification system. This changing regulatory framework has an impact on all stakeholders involved in the IVD Industry, including Authorized Representatives, Distributors, Importers, Notified Bodies, and Reference Laboratories and is expected to have a significant effect on the development of new CDx.

New design yields robust large-area framing camera.

Framing cameras provide a discrete series of images over a short period of time in a manner that mimics a high-speed movie camera but with individual shutter times that must be extremely short in order to capture freeze-frame images of rapidly evolving phenomena such as high-explosive shock-driven ejecta, dynamic compression of metals, and high-velocity fluid flow. The Nevada National Security Site has designed and fielded a new, large-area, gated framing camera called Kraken. The camera design emphasized manufacturability and flexibility by improving imager yield and creating a camera architecture to shorten design cycles. Design strategies and field data are presented.

PARP inhibitors and immunotherapy in ovarian and endometrial cancers.

Advanced ovarian and endometrial cancers have historically been associated with poor prognosis and few treatment options, limited to single or doublet chemotherapy regimens. The introduction of novel target therapies has transformed the management of these cancers. In contrast to chemotherapy, which inhibits DNA replication and mitosis, targeted therapies target cancer signalling pathways, stroma, immune-microenvironment and vasculature in tumour tissues. The most notable advances in gynaecological cancers have come from the introduction of PARP inhibitors and immune checkpoint inhibitors for ovarian and endometrial cancer, respectively. Several PARP inhibitors, which target defective DNA repair, have been approved as maintenance therapy for advanced ovarian cancer in both the first line and platinum-sensitive relapsed settings. Immune checkpoint inhibitors such as anti-PD-1/PD-L1 antibodies have proven successful in advanced mismatch repair deficient endometrial cancers with use now being investigated beyond this population. This review will explore the biological rationale and clinical evidence behind the use of PARP inhibitors and immunotherapy in ovarian and endometrial cancers.

Placement effects of inertial measurement units on contact identification in wheelchair racing.

Inertial measurement units (IMUs) provide a practical solution for attaining key performance data for wheelchair sports. The effects of IMU placement position on the identification of propulsion characteristics are unknown. The aim of this study was to determine the variability in the reliability of cycle time measurements (time between hand contacts) across IMU locations on the chair frame (axle housings), and wheels (axle, push rim, outer rim), on both the left and right sides (n = 8). Contacts were defined by spikes in the resultant acceleration data, corresponding to impact between the hands and push rim, and verified against motion capture. Five elite wheelchair racing athletes propelled at racing speeds on a treadmill. Excellent inter-rater Intraclass Correlation Coefficient values indicated high reliability and repeatability for both motion capture and IMU signal analysis approaches (R = 0.997, p < 0.001 and R = 0.990, p < 0.001, respectively). The best results were (as determined by the best between method agreement) were observed for IMUs located on the frame. Detection reliability was positively associated with signal-to-noise ratio of the acceleration data. The IMU assessment approach facilitates an automated processing capability, which is an improvement to the currently used video analysis.

A practical assessment of wheelchair racing performance kinetics using accelerometers.

Due to the detrimental influence of unnecessary mass on performance, racing wheelchair instrumentation used in both competition assessment and research is currently limited. Attaining key kinetic parameters of propulsion can enhance technique and provide athletes with a competitive advantage. This research examined the plausibility of inertial measurement units (IMUs) to estimate propulsion forces, during a simulated wheelchair race start and training. Start propulsion data calculated from an IMU system was compared to reference force plate data; steady state motion data was compared with existing literature. Some agreement in kinetic parameters between IMU data was observed under steady state motion, with data from athletes following a linear force-velocity relationship. In this context, it is important to identify that this cannot be directly compared to the existing literature due to the different methods of force measurement and the lack of data for similar force measurements using IMUs. IMUs were ineffective when used with wheelchairs having spoked wheels. Performance was best for measurements in the direction of motion. Although exact agreement was not observed, the IMU can provide an effective tool in the in-field assessment of propulsion kinetics.

Organ donation in the US and Europe: The supply vs demand imbalance.

Organ donation and transplantation remain the best and most cost-effective clinical solution for end-stage organ failure. Several agencies across the US and Europe provide legislative, regulatory, and humanitarian services to generate smoother applications in all transplantation processes and donor-recipient relationships. US and European statistics present nine types of grafts, with kidneys being the most transplanted organ worldwide. However, organ shortage, religion, underrepresented minority groups, difficulties in obtaining consent, lack of understanding, and general ethical concerns present challenging barriers to organ donation, reflecting the complexity of graft procurement and allocation. Breaking down these barriers to reduce the organ-supply imbalance requires an appropriate multifaceted approach. Some of the key areas include increasing the potential donor pool and consent rates, apt organ allocation, and improving organ health. Additionally, suitable policies and standardized guidelines for both donors and recipients, alongside educational initiatives, are needed to ensure patient safety and global awareness. Looking forward, novel and effective research plans and initiatives are needed if we are to avoid a colossal supply-demand gap.

If it's not one thing, HIF's another: immunoregulation by hypoxia inducible factors in disease.

Hypoxia-inducible factors (HIFs) have emerged in recent years as critical regulators of immunity. Localised, low oxygen tension is a hallmark of inflamed and infected tissues. Subsequent myeloid cell HIF stabilisation plays key roles in the innate immune response, alongside emerging oxygen-independent roles. Manipulation of regulatory proteins of the HIF transcription factor family can profoundly influence inflammatory profiles, innate immune cell function and pathogen clearance and, as such, has been proposed as a therapeutic strategy against inflammatory diseases. The direction and mode of HIF manipulation as a therapy are dictated by the inflammatory properties of the disease in question, with innate immune cell HIF reduction being, in general, advantageous during chronic inflammatory conditions, while upregulation of HIF is beneficial during infections. The therapeutic potential of targeting myeloid HIFs, both genetically and pharmacologically, has been recently illuminated in vitro and in vivo, with an emerging range of inhibitory and activating strategies becoming available. This review focuses on cutting edge findings that uncover the roles of myeloid cell HIF signalling on immunoregulation in the contexts of inflammation and infection and explores future directions of potential therapeutic strategies.