Using genetics and metabolomics we investigated the synthesis (gshA and gshB genes) and catabolism (ggt) of the conserved antioxidant glutathione in the model cyanobacterium Synechocystis PCC6803. These three genes are crucial to Synechocystis, in agreement with the proposed invention of glutathione by ancient cyanobacteria to protect themselves against the toxicity of oxygen they produced through photosynthesis. Consistent with their indispensability, gshA and gshB also operate in the production of another antioxidant, ergothioneine, as well as of the glutathione analogues ophthalmate and norophthalmate. Furthermore, we show that glutathione, ophthalmate and norophthalmate are accumulated in cells stressed by glucose, and that the two glutathione-dependent glyoxalase enzymes operate in the protection against glucose and its catabolite methylglyoxal. These findings are interesting because ophthalmate and norophthalmate were observed only in mammals so far, where ophthalmate is regarded as a biomarker of glutathione depletion. Instead, our data suggest that ophthalmate and norophthalmate are stress-induced markers of cysteine depletion triggered by its accelerated incorporation into glutathione, to face its increased demand for detoxification purposes. Hence, Synechocystis is an attractive model for the analysis of the role of glutathione, ergothioneine, ophthalmate and norophthalmate, in signalling and detoxification of oxidants and metabolic by-products.

OBJECTIVE: To evaluate the effects of Mind Over Matter: Healthy Bowels, Healthy Bladder, a small-group intervention, on urinary and bowel incontinence symptoms among older women with incontinence. METHODS: In this individually randomized group treatment trial, women aged 50 years and older with urinary, bowel incontinence, or both, were randomly allocated at baseline to participate in Mind Over Matter: Healthy Bowels, Healthy Bladder immediately (treatment group) or after final data collection (waitlist control group). The primary outcome was urinary incontinence (UI) improvement on the Patient Global Impression of Improvement at 4 months. Validated instruments assessed incontinence, self-efficacy, depression, and barriers to care-seeking. Intent-to-treat analyses compared differences between groups. Target sample size, based on an anticipated improvement rate of 45% in treated women vs 11% in the control group, 90% power, type I error of 0.05, with anticipated attrition of 25%, was 110. RESULTS: Among 121 women randomized (62 treatment group; 59 control group), 116 (95%) completed the 4-month assessment. Most participants were non-Hispanic white (97%), with a mean age of 75 years (SD 9.2, range 51–98); 66% had attended some college. There were no significant between-group differences at baseline. At 4 months, 71% of treated women vs 23% of women in the control group reported improved UI on Patient Global Impression of Improvement (P<.001); 39% vs 5% were much improved (P<.001). Regarding bowel incontinence, 55% of treated women vs 27% of women in the control group improved on Patient Global Impression of Improvement (P<.005), with 35% vs 11% reporting much improvement (P<.005). Treated women improved significantly more than women in the control group on all validated instruments of incontinence severity, quality of life, and self-efficacy. Care-seeking rates were similar between groups. CONCLUSION: Participation in a small-group intervention improves symptoms of both urinary and bowel incontinence in older women. Mind Over Matter is a feasible model with potential to bring effective behavioral solutions to the community. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, .

BackgroundUnrecognised changes in a hospitalised patient’s clinical course may lead to a preventable adverse event. Early warning systems (EWS) use patient data, such as vital signs, nursing assessments and laboratory values, to aid in the detection of early clinical deterioration. In 2018, an EWS programme was deployed at an academic hospital that consisted of a commercially available EWS algorithm and a centralised virtual nurse team to monitor alerts. Our objective was to understand the nursing perspective on the use of an EWS programme with centralised monitoring.MethodsWe conducted and audio-recorded semistructured focus groups during nurse staff meetings on six inpatient units, stratified by alert frequency (high: >100 alerts/month; medium: 50–100 alerts/month; low: <50 alerts/month). Discussion topics included EWS programme experiences, perception of EWS programme utility and EWS programme implementation. Investigators analysed the focus group transcripts using a grounded theory approach.ResultsWe conducted 28 focus groups with 227 bedside nurses across all shifts. We identified six principal themes: (1) Alert timeliness, nurses reported being aware of the patient’s deterioration before the EWS alert, (2) Lack of accuracy, nurses perceived most alerts as false positives, (3) Workflow interruptions caused by EWS alerts, (4) Questions of actionability of alerts, nurses were often uncertain about next steps, (5) Concerns around an underappreciation of core nursing skills via reliance on the EWS programme and (6) The opportunity cost of deploying the EWS programme.ConclusionThis qualitative study of nurses demonstrates the importance of earning user trust, ensuring timeliness and outlining actionable next steps when implementing an EWS. Careful attention to user workflow is required to maximise EWS impact on improving hospital quality and patient safety.

Skin models mimicking features of psoriasis‐related inflammation are needed to support the development of new drugs in dermatology. Reconstructed skin models lack tissue complexity, including a fully competent skin barrier, and presence and/or diversity of immune cells. Here, we describe InflammaSkin®, a novel human Th17‐driven ex vivo skin inflammation model. In this model, skin‐resident T cells are in situ activated by intradermal injection of anti‐CD3 and anti‐CD28 antibodies and Th17 cell polarization is sustained by culture in a chemically defined medium supplemented with IL‐1β, IL‐23 and TGF‐β for seven days. The acquired Th17 signature is demonstrated by the sustained secretion of IL‐17A, IL‐17AF, IL‐17F, IL‐22, IFN‐γ, and to some degree IL‐15 and TNF‐α observed in the activated ex vivo skin inflammation model compared with the non‐activated skin model control. Furthermore, expression of S100A7 and Keratin‐16 by keratinocytes and loss of epidermal structure integrity occur subsequently to in situ Th17cell activation, demonstrating cellular crosstalk between Th17 cells and keratinocytes. Finally, we demonstrate the use of this model to investigate the modulation of the IL‐23/IL‐17 immune axis by topically applied anti‐inflammatory compounds. Taken together, we show that by in situ activation of skin‐resident Th17 cells, the InflammaSkin® model reproduces aspects of inflammatory responses observed in psoriatic lesions and could be used as a translational tool to assess efficacy of test compounds.

Epidermal growth factor (EGF) is known to stimulate skin cell growth and synthesis of extracellular matrix such as hyaluronan(HA) which is decreased with aging in skin. However, EGF is hard to penetrate the stratum corneum and permeate into skin due to its size and hydrophilicity. To improve the skin delivery of EGF, we have used multi-lamellar complexes consist of EGF and 1,2-dioleoyl-3-tirmethylammonium-propane (DOTAP). EGF-DOTAP complex showed higher permeation rate in human skin and synthesis of hyaluronan in human skin equivalent model than normal liposome. A randomized, double-blind, vehiclecontrolled, split-face study was designed to evaluate the anti-aging effects of a cream containing EGF-DOTAP complex on human skin. Twentyeone Korean women completed a 12weeks study and were applied twice daily to the face. As compared with the vehicle control, EGF-DOTAP cream significantly improved the facial photo-damage score, skin roughness (R1), maximum roughness (R2), average roughness (R3, Ra), arithmetic roughness average (R5), and dermal density at 12 weeks. In conclusion, this study suggests that topical application of EGF-DOTAP complex exhibited significant improvement on skin aging by enhancing the permeation of EGF. LB1548 A pro-inflammatory environment modulates the human dermal fibroblast secretory phenotype: Implications for chronic wounds

Routine clinical assays, such as conventional immunohistochemistry, often fail to resolve the regional heterogeneity of complex inflammatory skin conditions. Here we introduce MANTIS (Multiplexed Annotated Tissue Imaging System), a flexible analytic pipeline compatible with routine practice, specifically-designed for spatially-resolved immune phenotyping of the skin in experimental or clinical samples. Based on phenotype attribution matrices coupled to α-shape algorithms, MANTIS projects a representative digital immune landscape, while enabling automated detection of major inflammatory clusters and concomitant single-cell data quantification of biomarkers. We observed that severe pathological lesions from systemic lupus erythematosus, Kawasaki syndrome or COVID-19-associated skin manifestations share common quantitative immune features, while displaying a non-random distribution of cells with the formation of disease-specific dermal immune structures. Given its accuracy and flexibility, MANTIS is designed to solve the spatial organization of complex immune environments to better apprehend the pathophysiology of skin manifestations.

To investigate the driving force behind the increase of ILCs in atopic dermatitis (AD) skin, we numerically and phenotypically compared ILC (sub)populations in the blood of AD patients and healthy controls (HC). For this purpose, peripheral blood mononuclear cells (PBMCs) were isolated from clinically active AD patients (n¼19) and age-matched (35.5 AE 0.1 years) healthy individuals (n¼17), immunostained with a panel of subset-characterising antibodies and analysed with a BD FACS Aria III sorter. Total ILCs were defined as viable CD45 + , CD3-, Lin-, CD127 + , CD161 + mononuclear cells and expressed as percent of CD45 + cells (mean AE SEM). Subpopulations of total ILCs were defined as follows: ILC1 (CD117-, CRTH2-), ILC2 (CRTH2 +), and ILC3 (CD117 + , CRTH2-). Results obtained show that the amount of total ILCs from PBMCs in AD patients is significantly lower than in healthy individuals (AD: 0.025 AE 0.004; HC: 0.044 AE 0.006, p ¼ 0.006). Comparing ILC subsets we observed significantly higher frequencies of ILC1 and lower percentages of ILC2 within total ILCs in AD versus healthy blood (ILC1: AD: 26.53 AE 4.63; HC: 12 AE 2.30; p ¼ 0.0104; ILC2: AD: 20.56 AE 2.78; HC: 34.32 AE 3.66; p ¼ 0.0046). No significant difference was observed for ILC3 in the blood of AD patients and HC. Our data support the concept that the reported increase of ILCs, particularly ILC2, in AD skin is at least partly due to emigration from blood and that this pathogenic loop could be a potential pharmacologic target.

Objectives-This study aimed to understand the potential reach of continence promotion intervention formats among incontinent women. Methods-The Survey of the Health of Wisconsin conducts household interviews on a population-based sample. In 2016, 399 adult women were asked about incontinence and likelihood of participation in continence promotion via 3 formats: single lecture, interactive 3-session workshop, or online. Descriptive analyses compared women likely versus unlikely to participate in continence promotion. To understand format preferences, modified grounded theory was used to conduct and analyze telephone interviews. Results-One hundred eighty-seven (76%) of 246 incontinent women reported being likely to attend continence promotion: 111 (45%) for a single lecture, 43 (17%) for an interactive 3-session workshop, and 156 (64%) for an online program. Obesity, older age, nonwhite race, prior health program participation, and Internet use for health information were associated with reported continence promotion participation. Cited advantages of a single lecture included convenience and ability to ask questions. A workshop offered accountability, hands-on learning, and opportunity to learn from others; online format offered privacy, convenience, and self-directed learning.