Umbelliferone alleviates impaired wound healing and skin barrier dysfunction in high glucose-exposed dermal fibroblasts and diabetic skins.

Skin wound healing is a complex process involving various cellular and molecular events. However, chronic wounds, particularly in individuals with diabetes, often experience delayed wound healing, potentially leading to diabetic skin complications. In this study, we examined the effects of umbelliferone on skin wound healing using dermal fibroblasts and skin tissues from a type 2 diabetic mouse model. Our results demonstrate that umbelliferone enhances several crucial aspects of wound healing. It increases the synthesis of key extracellular matrix components such as collagen I and fibronectin, as well as proteins involved in cell migration like EVL and Fascin-1. Additionally, umbelliferone boosts the secretion of angiogenesis factors VEGF and HIF-1α, enhances the expression of cell adhesion proteins including E-cadherin, ZO-1, and Occludin, and elevates levels of skin hydration-related proteins like HAS2 and AQP3. Notably, umbelliferone reduces the expression of HYAL, thereby potentially decreasing tissue permeability. As a result, it promotes extracellular matrix deposition, activates cell migration and proliferation, and stimulates pro-angiogenic factors while maintaining skin barrier functions. In summary, these findings underscore the therapeutic potential of umbelliferone in diabetic wound care, suggesting its promise as a treatment for diabetic skin complications. KEY MESSAGES: Umbelliferone suppressed the breakdown of extracellular matrix components in the skin dermis while promoting their synthesis. Umbelliferone augmented the migratory and proliferative capacities of fibroblasts. Umbelliferone activated the release of angiogenic factors in diabetic wounds, leading to accelerated wound healing. Umbelliferone bolstered intercellular adhesion and reinforced the skin barrier by preventing moisture loss and preserving skin hydration.

How Shift Nurses' Presenteeism is Related to Insomnia and Care Left Undone: A Cross-Sectional Study Using Generalised Structural Equation Modelling.

AIM: To examine the association between nurses' insomnia, presenteeism and care left undone and explore the mediating effect of presenteeism on the relationship between insomnia and care left undone among nurses in acute care hospitals, applying a dynamic model of presenteeism and absenteeism. DESIGN: A cross-sectional design. METHODS: A secondary data analysis was conducted using online survey data collected in January 2023. Participants were 1154 registered nurses who provided direct nursing care to patients in tertiary or general hospitals (hospitals with 300 or more beds) in South Korea. Poisson regression analysis was used to examine the association between nurses' insomnia, presenteeism and care left undone. Generalised structural equation modelling was performed to examine the mediating effect of presenteeism on the relationship between insomnia and care left undone. RESULTS: The prevalence of insomnia was 15.3%. More than half of the participants (63.7%) reported experiencing presenteeism in the past 4 weeks. All reported at least one nursing care left undone during their last shift. Insomnia and presenteeism were positively associated with care left undone, and presenteeism mediated the relationship between insomnia and care left undone among nurses in acute care hospitals. CONCLUSION: Nurses experienced insomnia and presenteeism, related to higher rates of care left undone. Nurses' insomnia had a direct effect on care left undone and an indirect effect through presenteeism. IMPLICATIONS FOR THE PROFESSION: Nurse managers and administrators need to make greater efforts to reduce insomnia and presenteeism among nurses, thereby reducing care left undone and improving patient safety in acute care settings. IMPACT: Nurse managers and administrators should consider proactive interventions to address nurses' insomnia and foster a workplace with healthy systems and culture to reduce sleep disturbance and presenteeism, which can effectively decrease care left undone. REPORTING METHOD: STROBE guidelines. PATIENT OR PUBLIC CONTRIBUTION: No patient or public contribution.

Job Characteristic Profiles and Presenteeism Among Shift Nurses: A Latent Profile Analysis Based on the Job Demands-Resources Model.

AIM(S): To investigate job characteristic profiles based on the job demands-resources (JD-R) model and the association between job characteristic profiles and presenteeism. DESIGN: Cross-sectional study. METHODS: Data were collected from shift nurses in South Korea in January 2023 using an online survey. This study included 544 nurses working in tertiary and general hospitals in South Korea. The participants were classified into four job characteristic profiles based on the JD-R model using latent profile analysis. Multiple logistic and linear regression analyses were performed to examine the association between job characteristic profiles and presenteeism. RESULTS: Four profiles were derived from the latent profile analysis: low strain-low motivation, high strain-low motivation, low strain-high motivation and high strain-high motivation. The high strain-low motivation, low strain-high motivation and high strain-high motivation groups were 1.74, 1.37 and 3.51 times more likely to experience presenteeism than the low strain-low motivation group respectively. The number of days worked while sick and the level of reduced job productivity was also the highest in the high strain-high motivation group. In addition, general health, multimorbidities and insomnia were significant factors affecting presenteeism, number of days worked while sick and productivity loss among shift nurses. CONCLUSIONS: This study found that even with higher levels of job resources, work environments with high levels of job demands were associated with higher rates of presenteeism. IMPACT: Study findings suggest that various strategies are needed to improve the physical and mental health of shift nurses, and create a healthy organisational environment by reducing excessive job demands to reduce presenteeism and increase productivity. REPORTING METHOD: STROBE guidelines. PATIENT OR PUBLIC CONTRIBUTION: No patient or public contribution.

Biological, Antifungal, and Physical Efficacy of a Denture Cleanser Formulated with Cnidium officinale Extracts.

BACKGROUND/OBJECTIVES: We aimed to assess the antifungal efficacy and impact of a denture cleanser containing Cnidium officinale extract on the surface characteristics of denture base materials, as well as its physical and biological properties. METHODS: The experimental denture cleansers were formulated with C. officinale at concentrations of 100 and 150 µg/mL, combined with 1% cocamidopropyl betaine as a natural surfactant. Antifungal efficacy was evaluated using zone-of-inhibition assays against Candida albicans, revealing inhibition zones of 20 ± 1.8 mm for the 100 µg/mL concentration and 23.6 ± 1.6 mm for the 150 µg/mL concentration. Surface property assessments-including hardness, roughness, color stability, and solubility measurements-demonstrated no significant differences compared to the control group. Biological evaluations included the quantification of polyphenol and flavonoid content. RESULTS: The C. officinale-based cleanser showed significant antifungal activity without affecting the hardness, roughness, color stability, or solubility of denture base materials. Biological tests revealed no cytotoxicity and minimal mucosal irritation. Polyphenol and flavonoid contents were quantitatively measured, revealing higher concentrations in the experimental groups, which were correlated with significant antifungal activity. These compounds are known for their roles in disrupting microbial processes and enhancing antimicrobial effects. These findings suggest that the C. officinale-based denture cleanser effectively inhibits C. albicans while preserving the physical properties of denture base materials. CONCLUSIONS: This study highlights the potential of C. officinale in denture cleanser formulations, promoting denture hygiene and oral health. Future research should prioritize long-term clinical evaluations and formulation optimization.

Impact of activity limitations due to fear of falling on changes in frailty in Korean older adults: a longitudinal study.

This study investigated the frailty change patterns among Korean older adults during 2006-2020 and the effect of activity limitations induced by the fear of falling (FOF) on these patterns. We employed a descriptive longitudinal design utilizing data from Waves 1 to 8 of the Korean Longitudinal Study of Aging. The exclusion criteria were a baseline age < 65 years, frailty index (FI) captured only at baseline, and death or unknown survival status. Multilevel modeling, combining regression equations at two levels, was used to examine the effect of activity limitations due to FOF on frailty, adjusting for other confounding factors. An increase in FI (1.97; p < .001) was demonstrated in participants who had experienced falls versus those who had not in the past 2 years. Notably, those facing activity limitations due to FOF exhibited a more significant increase in FI (4.62; p < .001) compared with those without; frailty progression intensified over time in the former (0.54, p < .001). Activity limitations due to FOF had a more pronounced impact on frailty than falls; moreover, these activity limitations accelerated the progression of frailty. Healthcare providers must prioritize addressing FOF by employing both physical and psychological interventions to mitigate activity limitations and ultimately decelerate frailty.

Delirium in neonates and infants: a scoping review protocol.

INTRODUCTION: Delirium in neonates and infants is difficult to screen or assess because of their different developmental features and distinct delirium characteristics compared with those of older children. Some delirium management strategies, including assessment, pharmacological and non-pharmacological interventions, and prevention strategies, have been previously suggested for paediatric delirium. However, whether these strategies are effective for delirium in neonates and infants is unclear. This scoping review aims to explore comprehensive information on delirium in neonates and infants, such as the features of delirium, factors related to delirium, and current assessments and interventions in neonates and infants. METHODS AND ANALYSIS: This study will be based on the Joanna Briggs Institute guidelines for scoping review protocol development and follow each stage of the framework proposed by Arksey and O'Malley. Research questions regarding delirium management in neonates and infants will be specified. A wide range of databases, including MEDLINE, EBSCO, Embase and the Cochrane Library, using optimised terms will be searched from 2004 to 2024. The summarised results will be reported in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension of the Scoping Reviews checklist. ETHICS AND DISSEMINATION: Since this study will review and summarise published scientific literature, ethical approval is not required. The results of this scoping review will be disseminated through conference presentations and peer-reviewed publications. The study findings will be disseminated through seminars for experts so that they can be reflected in practice.

A dispensable role of mural cell-derived laminin-α5 in intracerebral hemorrhage.

Although most laminin isoforms are neuroprotective in stroke, mural cell-derived laminin-α5 plays a detrimental role in an ischemia-reperfusion model. To determine whether this deleterious effect is an intrinsic feature of mural cell-derived laminin-α5 or unique to ischemic stroke, we performed loss-of-function studies using middle-aged mice with laminin-α5 deficiency in mural cells (α5-PKO) in an intracerebral hemorrhage (ICH) model. Control and α5-PKO mice exhibited comparable changes in all parameters examined, including hematoma size, neuronal death, neurological function, blood-brain barrier integrity, and reactive gliosis. These findings highlight a minimal role of mural cell-derived laminin-α5 in ICH. Together with the detrimental role of mural cell-derived laminin-α5 in ischemic stroke, these negative results in ICH model suggest that mural cell-derived laminin-α5 may exert distinct functions in different diseases.

High Throughput Correlative Electrochemistry-Microscopy Analysis on a Zn-Al Alloy.

Conventional electrodes and electrocatalysts possess complex compositional and structural motifs that impact their overall electrochemical activity. These motifs range from defects and crystal orientation on the electrode surface to layers and composites with other electrode components, such as binders. Therefore, it is vital to identify how these individual motifs alter the electrochemical activity of the electrode. Scanning electrochemical cell microscopy (SECCM) is a powerful tool that has been developed for investigating the electrochemical properties of complex structures. An example of a complex electrode surface is Zn-Al alloys, which are utilized in various sectors ranging from cathodic protection of steel to battery electrodes. Herein, voltammetric SECCM and correlative microstructure analysis are deployed to probe the electrochemical activities of a range of microstructural features, with 651 independent voltammetric measurements made in six distinctive areas on the surface of a Zn-Al alloy. Energy-dispersive X-ray spectroscopy (EDS) mapping reveals that specific phases of the alloy structure, particularly the α-phase Zn-Al, favor the early stages of metal dissolution (i.e., oxidation) and electrochemical reduction processes such as the oxygen reduction reaction (ORR) and redeposition of dissolved metal ions. A correlative analysis performed by comparing high-resolution quantitative elemental composition (i.e., EDS) with the corresponding spatially resolved cyclic voltammograms (i.e., SECCM) shows that the nanospot α-phase of the Zn-Al alloy contains high Al content (30-50%), which may facilitate local Al dissolution as the local pH increases during the ORR in unbuffered aqueous media. Overall, SECCM-based high-throughput electrochemical screening, combined with microstructure analysis, conclusively demonstrates that structure-composition heterogeneity significantly influences the local electrochemical activity on complex electrode surfaces. These insights are invaluable for the rational design of advanced electromaterials.

Eucalyptol Ameliorates Retinal Microvascular Defects through Modulating ER Stress and Angiopoietin-Tie Signaling in Diabetic Eyes.

Loss of the inner blood-retinal barrier (BRB) integrity is a main feature of ocular diseases such as diabetic macular edema. However, there is a lack of clarity on how inner BRB function is modulated within the diabetic retina. The current study examined whether eucalyptol inhibited inner BRB destruction and aberrant retinal angiogenesis in 33 mM glucose-exposed human retinal microvascular endothelial (RVE) cells and db/db mice. This study further examined the molecular mechanisms underlying endothelial dysfunction including retinal endoplasmic reticulum (ER) stress and angiopoietin (Ang)/Tie axis in conjunction with vascular endothelial growth factor (VEGF). Eucalyptol is a naturally occurring monoterpenoid and an achiral aromatic component of many plants including eucalyptus leaves. Nontoxic eucalyptol reduced the production of amyloid-ß (Aß) protein in glucose-loaded RVE cells and in diabetic mice. This natural compound blocked apoptosis of Aß-exposed RVE cells in diabetic mouse eyes by targeting ER stress via the inhibition of PERK-eIF2α-ATF4-CHOP signaling. Eucalyptol promoted activation of the Ang-1/Tie-2 pathway and dual inhibition of Ang-2/VEGF in Aß-exposed RVE cells and in diabetic eyes. Supply of eucalyptol reversed the induction of junction proteins in glucose/Aß-exposed RVE cells within the retina and reduced permeability. In addition, oral administration of eucalyptol reduced vascular leaks in diabetic retinal vessels. Taken together, these findings clearly show that eucalyptol inhibits glucose-induced Aß-mediated ER stress and manipulates Ang signaling in diabetic retinal vessels, which ultimately blocks abnormal angiogenesis and loss of inner BRB integrity. Therefore, eucalyptol provides new treatment strategies for diabetes-associated RVE defects through modulating diverse therapeutic targets including ER stress, Ang-1/Tie-2 signaling, and Ang-2/VEGF.

Five years of scanning electrochemical cell microscopy (SECCM): new insights and innovations.

Scanning electrochemical cell microscopy (SECCM) is a nanopipette-based technique which enables measurement of localised electrochemistry. SECCM has found use in a wide range of electrochemical applications, and due to the wider uptake of this technique in recent years, new applications and techniques have been developed. This minireview has collected all SECCM research articles published in the last 5 years, to demonstrate and celebrate the recent advances, and to make it easier for SECCM researchers to remain well-informed. The wide range of SECCM applications is demonstrated, which are categorised here into electrocatalysis, electroanalysis, photoelectrochemistry, biological materials, energy storage materials, corrosion, electrosynthesis, and instrumental development. In the collection of this library of SECCM studies, a few key trends emerge. (1) The range of materials and processes explored with SECCM has grown, with new applications emerging constantly. (2) The instrumental capabilities of SECCM have grown, with creative techniques being developed from research groups worldwide. (3) The SECCM research community has grown significantly, with adoption of the SECCM technique becoming more prominent.

Optimizing Amorphous Molybdenum Sulfide Thin Film Electrocatalysts: Trade-Off between Specific Activity and Microscopic Porosity.

Amorphous molybdenum sulfide (a-MoSx) is a promising candidate to replace noble metals as electrocatalysts for the hydrogen evolution reaction (HER) in electrochemical water splitting. So far, understanding of the activity of a-MoSx in relation to its physical (e.g., porosity) and chemical (e.g., Mo/S bonding environments) properties has mostly been derived from bulk electrochemical measurements, which provide limited information about electrode materials that possess microscopic structural heterogeneities. To overcome this limitation, herein, scanning electrochemical cell microscopy (SECCM) has been deployed to characterize the microscopic electrochemical activity of a-MoSx thin films (ca. 200 nm thickness), which possess a significant three-dimensional structure (i.e., intrinsic porosity) when produced by electrodeposition. A novel two-step SECCM protocol is designed to quantitatively determine spatially resolved electrochemical activity and electrochemical surface area (ECSA) within a single, high-throughput measurement. It is shown for the first time that although the highest surface area (e.g., most porous) regions of the a-MoSx film possess the highest total activity (measured by the electrochemical current), they do not possess the highest specific activity (measured by the ECSA-normalized current density). Instead, the areas of highest specific activity are localized at/around circular structures, coined "pockmarks", which are tens to hundreds of micrometers in size and ubiquitous to a-MoSx films produced by electrodeposition. By coupling this technique with structural and elemental composition analysis techniques (scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy) and correlating ECSA with activity and specific activity across SECCM scans, this work furthers the understanding of structure-activity relations in a-MoSx and highlights the importance of local measurements for the systematic and rational design of thin film catalyst materials.

Sleep Deprivation and Fatigue among Nurses Working Consecutive Night Shifts: A Prospective Observational Study.

PURPOSE: This study aimed to identify changes in sleep patterns and fatigue levels during consecutive night shifts among shift nurses and to determine the association between sleep parameters and increased fatigue levels during work. METHODS: This prospective observational study employing ecological momentary assessments was conducted using data collected from 98 shift nurses working in Korean hospitals between June 2019 and February 2021. The sleep patterns were recorded using actigraphy. The participants reported their fatigue levels at the beginning and end of each night shift in real time via a mobile link. Linear mixed models were used for the analysis. RESULTS: Nurses spent significantly less time in bed and had shorter sleep durations during consecutive night shifts than on off-duty days, whereas their wake times after sleep onset were much longer on off-duty days than on on-duty days. Fatigue levels were higher on the second and third night-shift days than on the first night-shift days. A shorter time spent in bed and asleep was associated with a greater increase in fatigue levels at the end of the shift than at the beginning. CONCLUSION: Nurses experience significant sleep deprivation during consecutive night shifts compared with off-duty days, and this sleep shortage is associated with a considerable increase in fatigue levels at the end of shifts. Nurse managers and administrators must ensure sufficient intershift recovery time during consecutive night shifts to increase the time spent in bed and sleeping.

Oligodendrocyte-derived laminin-γ1 regulates the blood-brain barrier and CNS myelination in mice.

Although oligodendrocytes (OLs) synthesize laminin-γ1, the most widely used γ subunit, its functional significance in the CNS remains unknown. To answer this important question, we generated a conditional knockout mouse line with laminin-γ1 deficiency in OL lineage cells (γ1-OKO). γ1-OKO mice exhibit weakness/paralysis and die by post-natal day 33. Additionally, they develop blood-brain barrier (BBB) disruption in the cortex and striatum. Subsequent studies reveal decreased major facilitator superfamily domain containing 2a expression and increased endothelial caveolae vesicles, but unaltered tight junction protein expression and tight junction ultrastructure, indicating a transcellular, rather than a paracellular, mechanism of BBB breakdown. Furthermore, significantly reduced OL lineage cells, OL precursor cells (OPCs), proliferating OPCs, and mature OLs are observed in γ1-OKO brains in a region-specific manner. Consistent with this finding, various defects in myelination are detected in γ1-OKO brains at biochemical and ultrastructural levels. Overall, these results highlight important roles of OL-derived laminin-γ1 in BBB maintenance and OL biology (proliferation, differentiation, and myelination).

Ellagic acid, a functional food component, ameliorates functionality of reverse cholesterol transport in murine model of atherosclerosis.

BACKGROUND/OBJECTIVES: High levels of plasma low-density lipoprotein (LDL) cholesterol are an important determinant of atherosclerotic lesion formation. The disruption of cholesterol efflux or reverse cholesterol transport (RCT) in peripheral tissues and macrophages may promote atherogenesis. The aim of the current study was to examine whether bioactive ellagic acid, a functional food component, improved RCT functionality and high-density lipoprotein (HDL) function in diet-induced atherogenesis of apolipoproteins E (apoE) knockout (KO) mice. MATERIALS/METHODS: Wild type mice and apoE KO mice were fed a high-cholesterol Paigen diet for 10 weeks to induce hypercholesterolemia and atherosclerosis, and concomitantly received 10 mg/kg ellagic acid via gavage. RESULTS: Supplying ellagic acid enhanced induction of apoE and ATP-binding cassette (ABC) transporter G1 in oxidized LDL-exposed macrophages, facilitating cholesterol efflux associated with RCT. Oral administration of ellagic acid to apoE KO mice fed on Paigen diet improved hypercholesterolemia with reduced atherogenic index. This compound enhanced the expression of ABC transporters in peritoneal macrophages isolated from apoE KO mice fed on Paigen diet, indicating increased cholesterol efflux. Plasma levels of cholesterol ester transport protein and phospholipid transport protein involved in RCT were elevated in mice lack of apoE gene, which was substantially reduced by supplementing ellagic acid to Paigen diet-fed mice. In addition, ellagic acid attenuated hepatic lipid accumulation in apoE KO mice, evidenced by staining of hematoxylin and eosin and oil red O. Furthermore, the supplementation of 10 mg/kg ellagic acid favorably influenced the transcriptional levels of hepatic LDL receptor and scavenger receptor-B1 in Paigen diet-fed apoE KO mice. CONCLUSION: Ellagic acid may be an athero-protective dietary compound encumbering diet-induced atherogenesis though improving the RCT functionality.

Practical guidelines for the use of scanning electrochemical cell microscopy (SECCM).

Scanning electrochemical cell microscopy (SECCM) has emerged as a transformative technology for electrochemical materials characterisation and the study of single entities, garnering global adoption by numerous research groups. While details on the instrumentation and operational principles of SECCM are readily available, the growing need for practical guidelines, troubleshooting strategies, and a systematic overview of applications and trends has become increasingly evident. This tutorial review addresses this gap by offering a comprehensive guide to the practical application of SECCM. The review begins with a discussion of recent developments and trends in the application of SECCM, before providing systematic approaches to (and the associated troubleshooting associated with) instrumental set up, probe fabrication, substrate preparation and the deployment of environmental (e.g., atmosphere and humidity) control. Serving as an invaluable resource, this tutorial review aims to equip researchers and practitioners entering the field with a comprehensive guide to essential considerations for conducting successful SECCM experiments.

Nursing experiences and knowledge of paediatric delirium: Analysing knowledge-practice gaps.

BACKGROUND: Delirium commonly occurs in paediatric patients with acute critical illness and negatively affects clinical outcomes. Variations in delirium knowledge levels and its management have been noted among nurses. AIMS: This study investigated nurses' experiences and knowledge levels regarding paediatric delirium. Additionally, we aimed to assess the gap between knowledge levels and practical experiences with paediatric delirium. STUDY DESIGN: This cross-sectional descriptive study conveniently sampled paediatric nurses from a university hospital in South Korea between September 2022 and March 2023. Nursing experiences with paediatric delirium and delirium knowledge levels were measured using structured survey questionnaires. Delirium knowledge was scored 0 to 47, and higher scores indicated higher levels of delirium-related knowledge. Data were analysed using descriptive statistics and presented as mean, standard deviation, frequency and percentage. RESULTS: A total of 127 paediatric nurses participated in this study; 40.2% had experience with 1-5 delirium cases in the previous year, and 86.6% (n = 110) had never used assessment tools for paediatric delirium assessment. The mean total delirium knowledge score was 34.45 ± 5.4; the mean scores of knowledge regarding aetiology, signs and symptoms and nursing management of delirium were 8.93 ± 1.31, 13.24 ± 2.81 and 12.3 ± 2.7, respectively. Interventions associated with a lower level of delirium-related knowledge and a lower performance rate included avoiding restraint use and maintaining hydration and electrolyte levels. Interventions associated with a higher level of delirium-related knowledge but a lower rate of performance comprised providing orientation, offering emotional support, allowing participants to stay with family members and administering medications to manage delirium. CONCLUSIONS: Although nurses working in paediatric units exhibited high rate of delirium nursing care, there was the gap between their delirium-related knowledge and practice. Nurses need to be aware of the insufficient part of delirium care, and paediatric delirium education should be reinforced. RELEVANCE TO CLINICAL PRACTICE: Preventing, assessing and systematically managing paediatric delirium is crucial, and considering the study results, delirium education among nurses is needed.

Endothelial and mural laminin-α5 contributes to neurovascular integrity maintenance.

BACKGROUND: Laminin-α5, a major component of the basal lamina, is predominantly synthesized by endothelial and mural cells (pericytes and vascular smooth muscle cells) in the CNS. Loss of laminin-α5 in either population fails to induce any abnormalities due to functional redundancy. Thus, the functional significance of laminin-α5 in neurovascular integrity remains unknown. Here, we hypothesize that ablation of laminin-α5 in both endothelial and mural cells increases neurovascular permeability. METHODS: The compound knockout mice were generated by crossing laminin-α5 floxed mice with Tie2-Cre and PDGFRß-Cre, which target endothelial cells and mural cells, respectively. Neurovascular permeability in these mutants was determined with both exogenous and endogenous tracers. Endothelial paracellular and transcellular permeability was assessed by examining the expression of tight junction proteins and transcytosis-associated proteins. In addition, transmission electron microscopy (TEM) was used to visualize tight junction ultrastructure and endothelial caveolae vesicles. Defects in pericytes and astrocytes were investigated by examining pericyte coverage/contact and astrocyte polarity. RESULTS: Elevated neurovascular permeability was observed in the mutants. Subsequent studies found increased Caveolin-1 and decreased major facilitator superfamily domain-containing protein 2a (MFSD2A) expression, but unaltered Claudin-5 or zonula occludens-1 (ZO-1) expression. Consistent with these results, mutant mice exhibited increased endothelial caveolae vesicle number with intact tight junction structure under TEM. Additionally, pericyte coverage and contact were also decreased in the mutant mice, while astrocyte polarity was unaffected. CONCLUSIONS: These results strongly indicate that endothelial and mural cell-derived laminin-α5 actively maintains neurovascular integrity via the transcellular rather than paracellular mechanism.

Prognostic Implication of Platelet Reactivity According to Left Ventricular Systolic Dysfunction Status in Patients Treated With Drug-Eluting Stent Implantation: Analysis of the PTRG-DES Consortium.

BACKGROUND: Coronary artery disease patients undergoing percutaneous coronary intervention (PCI) often exhibit reduced left ventricular ejection fraction (LVEF). However, the impact of LV dysfunction status in conjunction with platelet reactivity on clinical outcomes has not been previously investigated. METHODS: From the multicenter PTRG-DES (Platelet function and genoType-Related long-term prognosis in DES-treated patients) consortium, the patients were classified as preserved-EF (PEF: LVEF ≥ 50%) and reduced-EF (REF: LVEF< 5 0%) group by echocardiography. Platelet reactivity was measured using VerifyNow P2Y12 assay and high platelet reactivity (HPR) was defined as PRU ≥ 252. The major adverse cardiac and cerebrovascular events (MACCEs) were a composite of death, myocardial infarction, stent thrombosis and stroke at 5 years after PCI. Major bleeding was defined as Bleeding Academic Research Consortium bleeding types 3-5. RESULTS: A total of 13,160 patients from PTRG-DES, 9,319 (79.6%) patients with the results of both PRU and LVEF were analyzed. The incidence of MACCE and major bleeding was higher in REF group as compared with PEF group (MACCEs: hazard ratio [HR] 2.17, P < 0.001, 95% confidence interval [CI] 1.85-2.55; major bleeding: HR 1.78, P < 0.001, 95% CI 1.39-2.78). The highest rate of MACCEs was found in patients with REF and HPR, and the difference between the groups was statistically significant (HR 3.14 in REF(+)/HPR(+) vs. PEF(+)/HPR(-) group, P < 0.01, 95% CI 2.51-3.91). The frequency of major bleeding was not associated with the HPR in either group. CONCLUSION: LV dysfunction was associated with an increased incidence of MACCEs and major bleeding in patients who underwent PCI. The HPR status further exhibited significant increase of MACCEs in patients with LV dysfunction in a large, real-world registry. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT04734028.

A dispensable role of oligodendrocyte-derived laminin-α5 in brain homeostasis and intracerebral hemorrhage.

Laminin, a major component of the basal lamina in the CNS, is also expressed in oligodendrocytes (OLs). However, the function of OL-derived laminin remains largely unknown. Here, we performed loss-of-function studies using two OL-specific laminin-α5 conditional knockout mouse lines. Both mutants were grossly normal and displayed intact blood-brain barrier (BBB) integrity. In a mouse model of intracerebral hemorrhage (ICH), control mice and both mutants exhibited comparable hematoma size and neurological dysfunction. In addition, similar levels of hemoglobin and IgG leakage were detected in the mutant brains compared to the controls, indicating comparable BBB damage. Consistent with this finding, subsequent studies revealed no differences in tight junction protein (TJP) and caveolin-1 expression among control and knockout mice, suggesting that neither paracellular nor transcellular mechanism was affected in the mutants. Furthermore, compared to the controls, both mutant lines showed comparable oligodendrocyte number, oligodendrocyte proliferation rate, MBP/MAG levels, and SMI-32 expression, highlighting a minimal role of OL-derived laminin-α5 in OL biology. Together, these findings highlight a dispensable role of OL-derived laminin-α5 in both brain homeostasis and ICH pathogenesis.

Zinc Deficiency Promotes Calcification in Vascular Smooth Muscle Cells Independent of Alkaline Phosphatase Action and Partly Impacted by Pit1 Upregulation.

Inorganic phosphate (Pi) is a critical determinant of calcification, and its concentration is regulated by alkaline phosphatase (ALP) and Pit1. ALP is a key regulator of osteogenic calcification and acts by modulating local inorganic phosphate (Pi) concentrations through hydrolyzing pyrophosphate in the extracellular matrix (ECM). Pit1, a sodium-dependent phosphate transporter, regulates calcification via facilitating phosphate uptake within the cells. To investigate whether zinc differentially regulates osteoblastic and vascular calcifications, we examined ALP activity and Pit1 in osteoblastic and vascular smooth muscle cells (VSMCs). Our findings demonstrate that calcification in osteoblastic MC3T3-E1 cells is decreased via diminished ALP action under zinc deficiency. In contrast, zinc-deficiency-induced calcification in VSMCs is independent of ALP action, as demonstrated by very weak ALP activity and expression in calcified VSMCs. In zinc-deficient A7r5 VSMC, P accumulation increased with increasing Na phosphate concentration (3-7 mM) but not with ß-GP treatment, which requires ALP activity to generate Pi. Ca deposition also increased with Na phosphate in a dose-dependent manner; in contrast, ß-GP did not affect Ca deposition. In osteoblastic cells, Pit1 expression was not affected by zinc treatments. In contrast, Pit1 expression is highly upregulated in A7r5 VSMC under zinc deficiency. Using phosphonoformic acid, a competitive inhibitor of Pit1, we showed that calcification is inhibited in both A7r5 and MC3T3-E1 cells, indicating a requirement for Pit1 in both calcifications. Moreover, the downregulation of VSMC markers under zinc deficiency was restored by blocking Pit1. Taken together, our results imply that zinc-deficiency-induced calcification in VSMC is independent of ALP action in contrast to osteoblastic calcification. Moreover, Pit1 expression in VSMCs is a target for zinc deficiency and may mediate the inhibition of VSMC marker expression under zinc deficiency.