The host GTPase Dynamin 2 modulates apical junction structure to control cell-to-cell spread of Listeria monocytogenes.

The food-borne pathogen Listeria monocytogenes uses actin-based motility to generate plasma membrane protrusions that mediate the spread of bacteria between host cells. In polarized epithelial cells, efficient protrusion formation by L. monocytogenes requires the secreted bacterial protein InlC, which binds to a carboxyl-terminal Src homology 3 (SH3) domain in the human scaffolding protein Tuba. This interaction antagonizes Tuba, thereby diminishing cortical tension at the apical junctional complex and enhancing L. monocytogenes protrusion formation and spread. Tuba contains five SH3 domains apart from the domain that interacts with InlC. Here, we show that human GTPase Dynamin 2 associates with two SH3 domains in the amino-terminus of Tuba and acts together with this scaffolding protein to control the spread of L. monocytogenes. Genetic or pharmacological inhibition of Dynamin 2 or knockdown of Tuba each restored normal protrusion formation and spread to a bacterial strain deleted for the inlC gene (∆inlC). Dynamin 2 localized to apical junctions in uninfected human cells and protrusions in cells infected with L. monocytogenes. Localization of Dynamin 2 to junctions and protrusions depended on Tuba. Knockdown of Dynamin 2 or Tuba diminished junctional linearity, indicating a role for these proteins in controlling cortical tension. Infection with L. monocytogenes induced InlC-dependent displacement of Dynamin 2 from junctions, suggesting a possible mechanism of antagonism of this GTPase. Collectively, our results show that Dynamin 2 cooperates with Tuba to promote intercellular tension that restricts the spread of ∆inlC Listeria. By expressing InlC, wild-type L. monocytogenes overcomes this restriction.

Preventing Surgeon Work-Related Musculoskeletal Disorders: A Pilot Study of the Comprehensive Operating Room Ergonomics (CORE) Program.

IMPORTANCE: Work-related musculoskeletal disorders (WMSDs) among surgeons are markedly increasing. Several proposed interventions to reduce WMSDs among surgeons have been studied, but few follow an occupational therapy-oriented approach addressing biomechanical, psychophysical, and psychosocial risk factors. OBJECTIVE: To design, implement, and assess the potential of the Comprehensive Operating Room Ergonomics (CORE) program for surgeons, a holistic evidence-based ergonomics and wellness intervention grounded in occupational therapy principles. DESIGN: Mixed-methods pilot study with the quantitative strand embedded in the qualitative strand. SETTING: University-affiliated hospital. PARTICIPANTS: Six laparoscopic surgeons. OUTCOMES AND MEASURES: CORE program outcomes were assessed using qualitative and quantitative data to indicate changes in posture, physical discomfort, sense of wellness, and operating room (OR) ergonomic performance. The Rapid Upper Limb Assessment (RULA) was used to quantify surgeons' WMSD risk level before and after intervention. RESULTS: There were 12 baseline observations (two for each participant), and two or three post-CORE implementation observations. A statistically significant difference, F(1, 6) = 8.57, p = .03, was found between pre- and post-occupational therapy intervention RULA scores. Thematic analysis of surgeon feedback, which was overwhelmingly positive, identified five themes: postural alignment, areas of commonly reported physical pain or discomfort, setup of the OR environment, surgical ergonomics training, and ergonomics in everyday life. CONCLUSIONS AND RELEVANCE: The CORE program effectively decreased ergonomic risk factors to optimize surgeons' occupational performance in the OR. This study demonstrates a potential solution to how occupational therapists can holistically support surgeons and health care providers who are at risk for WMSDs. Plain-Language Summary: By 2025, a surgeon shortage is expected, partly because of the increase in surgeons' work-related musculoskeletal disorders, which affect their health and job continuity. This pilot study shows that the Comprehensive Operating Room Ergonomics program effectively addresses these problems. The study also serves as a framework for occupational therapy professionals to work with health care providers on ergonomics, benefiting population health. Results suggest that this approach could enhance surgeons' work conditions, supporting the American Occupational Therapy Association's Vision 2025 to improve health and quality of life.

Can We Screen for Limb Length Discrepancy on Spinal Radiographs of Patients With Adolescent Idiopathic Scoliosis?

BACKGROUND: The aim of this study was to determine whether height differences in the levels of the iliac crests and femoral heads on erect spinal radiographs can be used as indirect measurements for the screening and surveillance of limb length discrepancy (LLD) in patients with scoliosis. METHODS: Whole body posteroanterior (PA) and lateral erect radiographs of patients with adolescent idiopathic scoliosis (AIS) were retrospectively reviewed. Patients with congenital, syndromic, and neuromuscular scoliosis were excluded. A direct measurement of each limb was taken from the highest point of the femoral head to the middle of the tibial plafond; any difference between the sides was recorded as the LLD. In addition, the PACS Software tool was used to measure femoral head height difference (FHHD) and iliac crest height difference (ICHD). Pearson's correlation, linear regression, and Bland-Altman plots were used to determine the relationships between LLD and FHHD, and LLD and ICHD. RESULTS: Radiographs of 141 patients (92 women, 49 men) with an average age of 12.0±2.65 years were analyzed. A strong correlation (r=0.730, P<0.001) was found between LLD and FHHD; the correlation between LLD and ICHD was weaker (r=0.585, P<0.001). The Bland-Altman analysis showed good agreements of LLD with FHHD and ICHD. Linear regression analysis predicted an LLD of ≤10 mm based on an FHHD of ≤11.5 mm or an ICHD of ≤15.3 mm. CONCLUSIONS: FHHD and ICHD on spinal PA radiographs can be used for the screening and monitoring of LLD in patients with AIS with FHHD being the preferred indirect measurement. These measurements are readily learned and quick to perform. The FHHD and ICHD can be measured on any erect scoliosis PA radiograph. Therefore, these proxy measurements can be used to screen and monitor for LLD in patients with AIS. LEVEL OF EVIDENCE: Level III.

Mental Health Promotion in Hong Kong: A Service Evaluation of the Coolminds Pilot Study.

BACKGROUND: It is vital to improve the mental health (MH) of young people in Hong Kong, where the suicide rate has nearly doubled among those aged 15-29 years since 2014. This multi-method evaluation aimed to assess Coolminds, which promoted MH literacy by engaging students, parents, and educators in secondary schools in Hong Kong. METHODS: Students, parents, and teachers completed self-report measures before and after attending workshops aimed at raising MH literacy. Pre- and post-survey data were collected between November 2021 and August 2022. Qualitative data from participant feedback and a focus group interview with Coolminds staff were analyzed using descriptive content analysis. RESULTS: The final dataset included 2903 students, 395 parents, and 325 educators from 52 schools. Only 3 schools had engagement with all 3 key groups. Significant improvements in MH knowledge were observed after the Coolminds workshops for all 3 groups. Qualitative analysis identified a reactive approach to MH issues, and bureaucratic and logistical challenges as barriers to implementation. CONCLUSION: Findings support the effectiveness of the Coolminds curriculum in improving MH literacy for each group and the demand for MH promotions in school settings. The implementation challenges indicate the need for future initiatives to better align with local needs and practices.

Premature Activation of the HIV-1 Protease Is Influenced by Polymorphisms in the Hinge Region.

HIV-1 protease inhibitors are an essential component of antiretroviral therapy. However, drug resistance is a pervasive issue motivating a persistent search for novel therapies. Recent reports found that when protease activates within the host cell's cytosol, it facilitates the pyroptotic killing of infected cells. This has led to speculation that promoting protease activation, rather than inhibiting it, could help to eradicate infected cells and potentially cure HIV-1 infection. Here, we used a nanoscale flow cytometry-based assay to characterize protease resistance mutations and polymorphisms. We quantified protease activity, viral concentration, and premature protease activation and confirmed previous findings that major resistance mutations generally destabilize the protease structure. Intriguingly, we found evidence that common polymorphisms in the hinge domain of protease can influence its susceptibility to premature activation. This suggests that viral heterogeneity could pose a considerable challenge for therapeutic strategies aimed at inducing premature protease activation in the future.

An epigenetically distinct HSC subset supports thymic reconstitution.

Hematopoietic stem cells (HSCs) with multilineage potential are critical for effective T cell reconstitution and restoration of the adaptive immune system after allogeneic Hematopoietic Cell Transplantation (allo-HCT). The Kit lo subset of HSCs is enriched for multipotential precursors, 1, 2 but their T-cell lineage potential has not been well-characterized. We therefore studied the thymic reconstituting and T-cell potential of Kit lo HSCs. Using a preclinical allo-HCT model, we demonstrate that Kit lo HSCs support better thymic recovery, and T-cell reconstitution resulting in improved T cell responses to infection post-HCT. Furthermore, Kit lo HSCs with augmented BM lymphopoiesis mitigate age-associated thymic alterations, thus enhancing T-cell recovery in middle-aged hosts. We find the frequency of the Kit lo subset declines with age, providing one explanation for the reduced frequency of T-competent HSCs and reduced T-lymphopoietic potential in BM precursors of aged mice. 3, 4, 5 Chromatin profiling revealed that Kit lo HSCs exhibit higher activity of lymphoid-specifying transcription factors (TFs), including Zbtb1 . Deletion of Zbtb1 in Kit lo HSCs diminished their T-cell potential, while reinstating Zbtb1 in megakaryocytic-biased Kit hi HSCs rescued T-cell potential, in vitro and in vivo . Finally, we discover an analogous Kit lo HSC subset with enhanced lymphoid potential in human bone marrow. Our results demonstrate that Kit lo HSCs with enhanced lymphoid potential have a distinct underlying epigenetic program.

Experimental evolution under predation reduces body size in dung flies but courtship displays persist in males (Diptera: Sepsidae).

Exaggerated sexual traits, such as ornaments and courtship displays, are crucial for mate acquisition in many species and are often subject to directional runaway selection. However, in the face of high predation risk, natural selection can result in a reduction of conspicuous precopulatory displays to avoid detection by potential predators. Sexual selection may then favour increased investment in inconspicuous postcopulatory traits. Here, we investigated the transgenerational effects of predation on precopulatory male courtship and postcopulatory sexual traits (testes size, sperm length) in a dung fly, Sepsis punctum (Sepsidae). Behavioural assays prior to selection document a marked decrease in male courtship displays in the presence of a predator, the Asian Ant Mantis (Odontomantis planiceps). However, after ten generations of experimental evolution, flies exhibited a marked increase in courtship, both in the absence and presence of a predator. Additionally, under sustained predation pressure, male and female body size decreased but male postcopulatory traits were not significantly affected. These results suggest that precopulatory courtship can be under strong sexual selection even in the face of predation pressure. Larger flies were more susceptible to predation, and there could be canalisation of postcopulatory traits that are crucial for fertilisation.

Efficacy and Safety of WCFA19 (Weissella confusa WIKIM51) in Reducing Body Fat in Overweight and Obese Adults.

Background: WCFA19 (Weissella confusa WIKIM51), found during the fermentation of kimchi, is known for its inhibitory effects on body weight and body fat. This study looked at the impact of WCFA19 isolated from dandelion kimchi on weight loss in overweight and obese adults that are otherwise healthy. Methods: This study was conducted as a multicenter, double-blind, randomized, placebo-controlled study with 104 overweight and obese subjects. Subjects were randomized evenly into the test group (WCFA19, 500 mg, n = 40) or control group (n = 34) for 12 weeks from 14 June 2021 to 24 December 2021. Effects were based on DEXA to measure changes in body fat mass and percentage. Results: Among the 74 subjects analyzed, WCFA19 oral supplementation for 12 weeks resulted in a significant decrease in body fat mass of 633.38 ± 1396.17 g (p = 0.0066) in overweight and obese individuals in the experimental group. The control group showed an increase of 59.10 ± 1120.57 g (p = 0.7604), indicating a statistically significant difference between the two groups. There was also a statistically significant difference (p = 0.0448) in the change in body fat percentage, with a decrease of 0.41 ± 1.22% (p = 0.0424) in the experimental group and an increase of 0.17 ± 1.21% (p = 0.4078) in the control group. No significant adverse events were reported. Conclusions: Oral supplementation of 500 mg of WCFA19 for 12 weeks is associated with a decrease in body weight, particularly in body fat mass and percentage.

The Role of Human Voice in Direct-To-Consumer Health Communication.

Despite overall positive perceptions, many customers distrust direct-to-consumer genetic testing companies in regard to privacy and aftercare. Prior research suggests companies can assuage mistrust by personalizing communication when discussing sensitive health information. This content analysis of customer e-mails (N = 338) investigates the purpose, personalization, and use of the human voice strategy by direct-to-consumer genetic testing companies to gain trust. Results reveal that companies rarely use human voice when communicating with customers, most of the information provided is promotional, and they invite dialogue less over time. Theoretical and practical implications are provided.

Kalium channelrhodopsins effectively inhibit neurons.

The analysis of neural circuits has been revolutionized by optogenetic methods. Light-gated chloride-conducting anion channelrhodopsins (ACRs)-recently emerged as powerful neuron inhibitors. For cells or sub-neuronal compartments with high intracellular chloride concentrations, however, a chloride conductance can have instead an activating effect. The recently discovered light-gated, potassium-conducting, kalium channelrhodopsins (KCRs) might serve as an alternative in these situations, with potentially broad application. As yet, KCRs have not been shown to confer potent inhibitory effects in small genetically tractable animals. Here, we evaluated the utility of KCRs to suppress behavior and inhibit neural activity in Drosophila, Caenorhabditis elegans, and zebrafish. In direct comparisons with ACR1, a KCR1 variant with enhanced plasma-membrane trafficking displayed comparable potency, but with improved properties that include reduced toxicity and superior efficacy in putative high-chloride cells. This comparative analysis of behavioral inhibition between chloride- and potassium-selective silencing tools establishes KCRs as next-generation optogenetic inhibitors for in vivo circuit analysis in behaving animals.

Exploring the personal stroke and rehabilitation experiences of older adults with chronic stroke during the COVID-19 pandemic: a qualitative descriptive study.

PURPOSE: The purpose of this study was to explore the personal stroke and rehabilitation experiences of older adults with chronic stroke living in a mid-sized Northwestern Ontario city in Canada during the COVID-19 pandemic. METHODS: A qualitative descriptive approach with a constructivist worldview was used. In addition, a semi-structured interview guide was used to gather the participants' perspectives on their experiences throughout stroke recovery. Ten participants were interviewed, including six males and four females. The interviews were completed, transcribed, and analysed using inductive and deductive content analysis. Multiple steps were taken to enhance data trustworthiness. RESULTS: Six main themes and eight related subthemes emerged. These included: getting help is complex, the effects of stroke are multifaceted, losing rehabilitation services during the COVID-19 pandemic, overcoming hardships but not alone, "If you don't use it, you lost it": rehabilitative success is based on one's actions, and "look at me now": the importance of taking pride in one's successes. CONCLUSIONS: One unique finding was that the participants used this study as an opportunity to teach and advocate for future stroke survivors which is not often seen in qualitative stroke rehabilitation research. Future stroke research should place emphasis on both the positive and negative experiences of this population.

Can the Lower Extremity Functional Scale Be Used in Children and Adolescents? A Validation Study.

BACKGROUND: The lower extremity functional scale (LEFS) is a patient-reported outcome measure for assessment of lower extremity function. It has been validated in adults but not in children or adolescents. METHODS: Patients 8 to 18 years of age who were treated for a lower limb fracture, injury, or other conditions were invited to join the study. LEFS and Pediatric Quality of Life Inventory 4.0 Generic Core Scales (PedsQL) were administered. Reliability and validity of the LEFS were determined for the entire cohort and for 2 age groups (children: 8 to 12 years; adolescents 13 to 18 years) using PedsQL as comparison. RESULTS: A total of 178 patients were recruited into this study. In the entire cohort, internal consistency of LEFS was excellent (0. 972) with acceptable floor (0%) and ceiling (12%) effects. Correlation between LEFS and PedsQL physical functioning component was high ( r =0.859). Construct validity was acceptable, with all 8 hypotheses demonstrating statistical significance. Factor analysis showed that item 15 (sitting for 1 hour) may contribute to measurement error in the pediatric population. Results remained similar when comparing the 2 age groups. CONCLUSIONS: The LEFS is overall an acceptable patient-reported outcome assessment of children and adolescents with various lower limbs disorders. LEVEL OF EVIDENCE: Level II.

A pilot study on the "ScoliBrace" in the treatment of adolescent idiopathic scoliosis.

PURPOSE: Bracing for adolescents with idiopathic scoliosis (AIS) is a treatment option to prevent curve progression to surgical level. This study aimed to assess the efficacy of a 3D fully customized over corrective brace, "ScoliBrace," an orthosis treatment for AIS. METHODS: This was a prospective pilot study of AIS female patients with inclusion criteria followed recommended Scoliosis Research Society (SRS) Guidelines. Cobb angles measured at: baseline (T0), 21 months (T5-2), skeletal maturity (T6), 6 months post-brace (T7), along with hours of brace wear using a thermal sensor and health-related quality of life (HRQoL) using the SRS-22r questionnaire. RESULTS: A total of 30 female AIS patients with mean age 11.85 ± 0.68 years, predominantly Risser 0 (70%), and median Cobb angle 29° were recruited; 21 patients were included for the final analysis. Results showed significant difference in Cobb angle between T0 and T5-2 (median = 22.5° vs. 28.5°, p = 0.0082). 57.14% had reduction in Cobb angle by ≥ 5° at skeletal maturity. Cobb angle reduced 0.794° for each additional hour of dosage (p = 0.036, 95% CI = - 1.532°, - 0.056°). Although pain level was increased at T6 (4.37 ± 0.51vs.4.70 ± 0.41, p = 0.014), patients reported significantly greater satisfaction with management of their condition (3.90 ± 0.90vs.3.29 ± 0.88, p = 0.020). CONCLUSION: Results show similar findings to the BRAIST study, whereby curves remained under surgical threshold and showed improvement. More than half had curve reduction of ≥ 5° at skeletal maturity. Increased dose was also associated with improved outcomes. Using "ScoliBrace" as a non-surgical treatment, maintained curves below surgical threshold and showed curve reduction, improving patient satisfaction with management.

Systematic review of paediatric pulseless pink humerus supracondylar fractures.

The supracondylar humeral fracture (SCHF) is the most common fracture seen in children, forming up to 70% of all pediatric elbow fractures. The decision to surgically explore the brachial artery in a well-perfused, pulseless arm remains a controversial one among vascular and orthopedic surgeons and is something we seek to explore in this article. We reviewed the literature from electronic databases such as PubMed and Embase for studies focusing on the management of the pink pulseless hand (PPH) following SCHF. We gathered a total of 23 articles to be analyzed in this review. We found 336 PPH postreduction and evaluated the management as well as the follow-up and complications of the PPH involved. Most recent articles have cited the close observation strategy as the most reliable strategy. It is clear that the management of a SCHF requires immediate reduction and fixation. In the management of a postoperatively pulseless pink humerus, we do agree with the latest conclusion of Delionitis et al . who advocate the traditional dogma of watchful waiting in the case of a PPH postreduction and fixation as long as no signs of vascular deterioration appear. However, the outcome of this study also advocates for the monitoring of up to 24-48 h postreduction and the use of noninvasive tools such as ultrasound Doppler, Color flow duplex Ultrasound and pulse oximetry to monitor perfusion as they all have had good outcomes in the articles cited in this review. Still, in the event of vascular deterioration or development of complications of the hand, immediate vascular exploration is still indicated.