Correlative light and X-ray tomography jointly unveil the critical role of connexin43 channels on inflammation-induced cellular ultrastructural alterations.

Non-junctional connexin43 (Cx43) plasma membrane hemichannels have been implicated in several inflammatory diseases, particularly playing a role in ATP release that triggers activation of the inflammasome. Therapies targeting the blocking of the hemichannels to prevent the pathological release or uptake of ions and signalling molecules through its pores are of therapeutic interest. To date, there is no close-to-native, high-definition documentation of the impact of Cx43 hemichannel-mediated inflammation on cellular ultrastructure, neither is there a robust account of the ultrastructural changes that occur following treatment with selective Cx43 hemichannel blockers such as Xentry-Gap19 (XG19). A combination of same-sample correlative high-resolution three-dimensional fluorescence microscopy and soft X-ray tomography at cryogenic temperatures, enabled in the identification of novel 3D molecular interactions within the cellular milieu when comparing behaviour in healthy states and during the early onset or late stages under inflammatory conditions. Notably, our findings suggest that XG19 blockage of connexin hemichannels under pro-inflammatory conditions may be crucial in preventing the direct degradation of connexosomes by lysosomes, without affecting connexin protein translation and trafficking. We also delineated fine and gross cellular phenotypes, characteristic of inflammatory insult or road-to-recovery from inflammation, where XG19 could indirectly prevent and reverse inflammatory cytokine-induced mitochondrial swelling and cellular hypertrophy through its action on Cx43 hemichannels. Our findings suggest that XG19 might have prophylactic and therapeutic effects on the inflammatory response, in line with functional studies.

Feasibility of linking universal child and family healthcare and financial counselling: findings from the Australian Healthier Wealthier Families (HWF) mixed-methods study.

OBJECTIVES: 'Healthier Wealthier Families' (HWF) seeks to reduce financial hardship in the early years by embedding a referral pathway between Australia's universal child and family health (CFH) services and financial counselling. This pilot study investigated the feasibility and short-term impacts of HWF, adapted from a successful Scottish initiative. METHODS: Setting: CFH services in five sites across two states, coinciding with the COVID-19 pandemic. PARTICIPANTS: Caregivers of children aged 0-5 years experiencing financial hardship (study-designed screen). DESIGN: Mixed methods. With limited progress using a randomised trial (RCT) design in sites 1-3 (March 2020-November 2021), qualitative interviews with service providers identified implementation barriers including stigma, lack of knowledge of financial counselling, low financial literacy, research burden and pandemic disruption. This informed a simplified RCT protocol (site 4) and direct referral model (no randomisation, pre-post evaluation, site 5) (June 2021-May 2022). INTERVENTION: financial counselling; comparator: usual care (sites 1-4). Feasibility measures: proportions of caregivers screened, enrolled, followed up and who accessed financial counselling. Impact measures: finances (quantitative) and other (qualitative) to 6 months post-enrolment. RESULTS: 355/434 caregivers completed the screen (60%-100% across sites). In RCT sites (1-4), 79/365 (19%-41%) reported hardship but less than one-quarter enrolled. In site 5, n=66/69 (96%) caregivers reported hardship and 44/66 (67%) engaged with financial counselling; common issues were utility debts (73%), and obtaining entitlements (43%) or material aid/emergency relief (27%). Per family, financial counselling increased income from government entitlements by an average $A6504 annually plus $A784 from concessions, grants, brokerage and debt waivers. Caregivers described benefits (qualitative) including reduced stress, practical help, increased knowledge and empowerment. CONCLUSIONS: Financial hardship screening via CFH was acceptable to caregivers, direct referral was feasible, but individual randomisation was infeasible. Larger-scale implementation will require careful, staged adaptations where CFH populations and the intervention are well matched and low burden evaluation. TRIAL REGISTRATION NUMBER: ACTRN12620000154909.

Density and distribution of dendritiform cells in the peripheral cornea of healthy subjects using in vivo confocal microscopy.

PURPOSE: To establish in a large healthy cohort, dendritiform cell (DC) density and morphological parameters in the central and peripheral cornea using in vivo confocal microscopy (IVCM). METHODS: A prospective, cross-sectional, observational study was conducted in 85 healthy volunteers (n = 85 eyes). IVCM images of corneal center and four peripheral zones were analyzed for DC density and morphology to compare means and assess correlations (p < 0.05 being statistically significant). RESULTS: Central corneas had lower DC density (40.83 ± 5.14 cells/mm2; mean ± SEM) as compared to peripheral corneas (75.42 ± 2.67 cells/mm2, p < 0.0001). Inferior and superior zones demonstrated higher DC density (105.01 ± 7.12 and 90.62 ± 4.62 cells/mm2) compared to the nasal and temporal zones (59.93 ± 3.42 and 51.77 ± 2.98 cells/mm2, p < 0.0001). Similarly, lower DC size, field and number of dendrites were observed in the central as compared to the average peripheral cornea (p < 0.0001), with highest values in the inferior zone (p < 0.001 for all, except p < 0.05 for number of dendrites in superior zone). DC parameters did not correlate with age or gender. Inter-observer reliability was 0.987 for DC density and 0.771-0.922 for morphology. CONCLUSION: In healthy individuals, the peripheral cornea demonstrates higher DC density and larger morphology compared to the center, with highest values in the inferior zone. We provide the largest normative cohort for sub-stratified DC density and morphology, which can be used in future clinical trials to compare differential changes in diseased states. Furthermore, as DC parameters in the peripheral zones are dissimilar, random sampling of peripheral cornea may be inaccurate.

Isothiourea-Catalyzed Enantioselective Michael Addition of Malonates to α,ß-Unsaturated Aryl Esters.

An enantioselective Michael addition of malonates to α,ß-unsaturated para-nitrophenyl esters was achieved using the Lewis basic isothiourea HyperBTM, giving excellent levels of product enantioselectivity (up to >99:1 enantiomeric ratio) in good yields and with complete regioselectivity (>20:1 regioselectivity ratio) in the presence of alternative (phenyl ketone and ethyl ester) Michael acceptors. Density functional theory calculations indicate that N-acylation is rate-limiting. This constitutes a rare example of a highly enantioselective addition of simple, readily available malonates to α,ß-unsaturated esters.

Twitchy, the Drosophila orthologue of the ciliary gating protein FBF1/dyf-19, is required for coordinated locomotion and male fertility.

Primary cilia are compartmentalised from the rest of the cell by a ciliary gate comprising transition fibres and a transition zone. The ciliary gate allows the selective import and export of molecules such as transmembrane receptors and transport proteins. These are required for the assembly of the cilium, its function as a sensory and signalling centre and to maintain its distinctive composition. Certain motile cilia can also form within the cytosol as exemplified by human and Drosophila sperm. The role of transition fibre proteins has not been well described in the cytoplasmic cilia. Drosophila have both compartmentalised primary cilia, in sensory neurons, and sperm flagella that form within the cytosol. Here, we describe phenotypes for twitchy the Drosophila orthologue of a transition fibre protein, mammalian FBF1/C. elegans dyf-19. Loss-of-function mutants in twitchy are adult lethal and display a severely uncoordinated phenotype. Twitchy flies are too uncoordinated to mate but RNAi-mediated loss of twitchy specifically within the male germline results in coordinated but infertile adults. Examination of sperm from twitchy RNAi-knockdown flies shows that the flagellar axoneme forms, elongates and is post-translationally modified by polyglycylation but the production of motile sperm is impaired. These results indicate that twitchy is required for the function of both sensory cilia that are compartmentalised from the rest of the cell and sperm flagella that are formed within the cytosol of the cell. Twitchy is therefore likely to function as part of a molecular gate in sensory neurons but may have a distinct function in sperm cells.

Inpatient Virtual Vision Clinic Improves Access to Vision Rehabilitation Before and During the COVID-19 Pandemic.

OBJECTIVE: To describe and evaluate a secure video call system combined with a suite of iPad vision testing apps to improve access to vision rehabilitation assessment for inpatients. DESIGN: Retrospective. SETTING: Two acute care inpatient rehabilitation hospitals and 1 long-term acute care (LTAC) hospital. PARTICIPANTS: Records of inpatients seen by the vision service. INTERVENTIONS: Records from a 1-year telemedicine pilot performed at acute rehabilitation (AR) hospital 1 and then expanded to AR hospital 2 and LTAC hospital during coronavirus disease 2019 (COVID-19) were reviewed. In the virtual visits, an occupational therapist measured the patients' vision with the iPad applications and forwarded results to the off-site Doctor of Optometry (OD) for review prior to a video visit. The OD provided diagnosis and education, press-on prism application supervision, strategies and modifications, and follow-up recommendations. Providers completed the telehealth usability questionnaire (10-point scale). MAIN OUTCOME MEASURES: Vision examinations per month at AR hospital 1 before and with telemedicine. RESULTS: With telemedicine at AR hospital 1, mean visits per month significantly increased from 10.7±5 to 14.9±5 (P=.002). Prism was trialed in 40% of cases of which 83% were successful, similar to previously reported in-person success rates. COVID-19 caused only a marginal decrease in visits per month (P=.08) at AR1, whereas the site without an established program (AR hospital 2) had a 3-4 week gap in care while the program was initiated. Cases at the LTAC hospital tended to be more complex and difficult to manage virtually. The telehealth usability questionnaire median category scores were 7 for Ease of Use, 8 for Interface Quality, 6 for Reliability, and 9 for Satisfaction and Future Use. CONCLUSIONS: The virtual vision clinic process improved inpatient access to eye and visual neurorehabilitation assessment before and during the COVID-19 quarantine and was well accepted by providers and patients.

In vivo measurement of shear modulus of the human cornea using optical coherence elastography.

Corneal stiffness plays a critical role in shaping the cornea with respect to intraocular pressure and physical interventions. However, it remains difficult to measure the mechanical properties noninvasively. Here, we report the first measurement of shear modulus in human corneas in vivo using optical coherence elastography (OCE) based on surface elastic waves. In a pilot study of 12 healthy subjects aged between 25 and 67, the Rayleigh-wave speed was 7.86 ± 0.75 m/s, corresponding to a shear modulus of 72 ± 14 kPa. Our data reveal two unexpected trends: no correlation was found between the wave speed and IOP between 13-18 mmHg, and shear modulus decreases with age (- 0.32 ± 0.17 m/s per decade). We propose that shear stiffness is governed by the interfibrillar matrix, whereas tensile strength is dominated by collagen fibrils. Rayleigh-wave OCE may prove useful for clinical diagnosis, refractive surgeries, and treatment monitoring.

Lunate fractures and associated radiocarpal and midcarpal instabilities: a systematic review.

The aim of this study was to analyse lunate fractures and any associated osseo-ligamentous injuries. A systematic review identified 34 cases. We identified carpal instabilities at the radiocarpal and midcarpal joints in volar and dorsal directions. Radiocarpal instabilities (10/34) were usually dorsoradial (8/10), with a transverse lunate fracture, best seen on a coronal image. Midcarpal instabilities (24/34) were usually volar (14/18), with a volar lunate shear fracture, best seen on a sagittal image. Instabilities were sub-classified into non-displaced, subluxated and dislocated. Associated fractures of the scaphoid and the radial and ulnar styloid processes were common. Lunate fractures without subluxation or dislocation had good outcomes with cast immobilization or fixation of associated fractures. Lunate fracture-subluxations are unstable injuries that are best managed with fixation of the carpal fractures. Lunate fracture-dislocations are complex injuries, requiring stabilization of the lunate, associated fractures and ligament injuries; complications are common and acute or delayed salvage procedures may be required.

Classroom Promotion of Oral Language (CPOL): protocol for a cluster randomised controlled trial of a school-based intervention to improve children's literacy outcomes at grade 3, oral language and mental health.

INTRODUCTION: Oral language and literacy competence are major influences on children's developmental pathways and life success. Children who do not develop the necessary language and literacy skills in the early years of school then go on to face long-term difficulties. Improving teacher effectiveness may be a critical step in lifting oral language and literacy outcomes. The Classroom Promotion of Oral Language trial aims to determine whether a specifically designed teacher professional learning programme focusing on promoting oral language can lead to improved teacher knowledge and practice, and advance outcomes in oral language and literacy for early years school children, compared with usual practice. METHODS AND ANALYSIS: This is a two-arm cluster multisite randomised controlled trial conducted within Catholic and Government primary schools across Victoria, Australia. The intervention comprises 4 days of face-to-face professional learning for teachers and ongoing implementation support via a specific worker. The primary outcome is reading ability of the students at grade 3, and the secondary outcomes are teacher knowledge and practice, student mental health, reading comprehension and language ability at grade 1; and literacy, writing and numeracy at grade 3. Economic evaluation will compare the incremental costs of the intervention to the measured primary and secondary outcomes. ETHICS AND DISSEMINATION: This trial was approved by the Monash University Human Research Ethics Committee #CF13/2634-2013001403 and later transferred to the University of Melbourne #1545540. The investigators (including Government and Catholic partners) will communicate trial results to stakeholders, collaborators and participating schools and teachers via appropriate presentations and publications. TRIAL REGISTRATION NUMBER: ISRCTN77681972; Pre-results.

Shoulder impingement syndrome: a systematic review of clinical trial participant selection criteria.

BACKGROUND: Shoulder impingement syndrome (SIS) is a common diagnosis for patients with pain and dysfunction of the shoulder. Variations in the signs and symptoms might lead to uncertainty regarding the definition of SIS. The aim of this review is to explore the participant selection criteria used in the literature when investigating SIS and to assess differences in criteria among treating professions. METHODS: This is a PRISMA systematic review of publications from 2009 to 2014 from MEDLINE, PubMed, The Cochrane Library, Embase, Scopus and CINAHL. RESULTS: Ninety-seven articles met inclusion criteria for this review. Twenty-five different surgical and nonsurgical treatments were investigated. Impingement-specific index tests were used in all studies. Exclusion index tests were used in 62% of studies. Twenty index tests were identified. Radiological investigations were reported in 53% of all studies, of which a further 53% reported using two or more radiological investigations. CONCLUSIONS: This systematic review has illustrated that studies investigating SIS test for various signs and symptoms, which is in keeping with describing the condition as a 'syndrome'. However, there are inconsistencies in participant selection criteria between health disciplines, highlighting a need for harmonization of the selection criteria in the form of an international editorial consensus.

Language skills of children during the first 12 months after stuttering onset.

PURPOSE: To describe the language development in a sample of young children who stutter during the first 12 months after stuttering onset was reported. METHODS: Language production was analysed in a sample of 66 children who stuttered (aged 2-4 years). The sample were identified from a pre-existing prospective, community based longitudinal cohort. Data were collected at three time points within the first year after stuttering onset. Stuttering severity was measured, and global indicators of expressive language proficiency (length of utterances and grammatical complexity) were derived from the samples and summarised. Language production abilities of the children who stutter were contrasted with normative data. RESULTS: The majority of children's stuttering was rated as mild in severity, with more than 83% of participants demonstrating very mild or mild stuttering at each of the time points studied. The participants demonstrated developmentally appropriate spoken language skills comparable with available normative data. CONCLUSION: In the first year following the report of stuttering onset, the language skills of the children who were stuttering progressed in a manner that is consistent with developmental expectations.

Latanoprost-Eluting Contact Lenses in Glaucomatous Monkeys.

PURPOSE: To assess the ability of latanoprost-eluting contact lenses to lower the intraocular pressure (IOP) of glaucomatous eyes of cynomolgus monkeys. DESIGN: Preclinical efficacy study of 3 treatment arms in a crossover design. PARTICIPANTS: Female cynomolgus monkeys with glaucoma induced in 1 eye by repeated argon laser trabeculoplasty. METHODS: Latanoprost-eluting low-dose contact lenses (CLLO) and high-dose contact lenses (CLHI) were produced by encapsulating a thin latanoprost-polymer film within the periphery of a methafilcon hydrogel, which was lathed into a contact lens. We assessed the IOP-lowering effect of CLLO, CLHI, or daily latanoprost ophthalmic solution in the same monkeys. Each monkey consecutively received 1 week of continuous-wear CLLO, 3 weeks without treatment, 5 days of latanoprost drops, 3 weeks without treatment, and 1 week of continuous-wear CLHI. On 2 consecutive days before initiation of each study arm, the IOP was measured hourly over 7 consecutive hours to establish the baseline IOP. Two-tailed Student t tests and repeated-measures analysis of variance were used for statistical analysis. MAIN OUTCOME MEASURES: Intraocular pressure. RESULTS: Latanoprost ophthalmic solution resulted in IOP reduction of 5.4±1.0 mmHg on day 3 and peak IOP reduction of 6.6±1.3 mmHg on day 5. The CLLO reduced IOP by 6.3±1.0, 6.7±0.3, and 6.7±0.3 mmHg on days 3, 5, and 8, respectively. The CLHI lowered IOP by 10.5±1.4, 11.1±4.0, and 10.0±2.5 mmHg on days 3, 5, and 8, respectively. For the CLLO and CLHI, the IOP was statistically significantly reduced compared with the untreated baseline at most time points measured. The CLHI demonstrated greater IOP reduction than latanoprost ophthalmic solution on day 3 (P = 0.001) and day 5 (P = 0.015), and at several time points on day 8 (P < 0.05). CONCLUSIONS: Sustained delivery of latanoprost by contact lenses is at least as effective as delivery with daily latanoprost ophthalmic solution. More research is needed to determine the optimal continuous-release dose that would be well tolerated and maximally effective. Contact lens drug delivery may become an option for the treatment of glaucoma and a platform for ocular drug delivery.

Estimating the Prevalence of Sleep-Disordered Breathing Among Collegiate Football Players.

BACKGROUND: Obstructive sleep apnea is a clinical disorder characterized by loud snoring, apneic episodes, and chronic sleep disruption. Collegiate football players exhibit several risk factors for OSA, including large neck circumference and high body mass index, although the prevalence of OSA in this cohort is unknown. METHODS: The STOP-BANG questionnaire was administered at random to members of a collegiate football team and used to stratify the players into high and low risk for sleep-disordered breathing (SDB). Those who completed the questionnaire were then evaluated for SDB during preseason camp using a single-channel (finger pulse oximetry) photoplethysmography-based device. SDB was defined as an apnea-hypopnea index of ≥5. RESULTS: Of 56 players who underwent overnight photoplethysmography monitoring, valid results were available for 51. Forty-eight percent of the players were high-risk (neck size = 44.6 ± 2.2 cm, body mass index = 33.0 ± 5.4) versus low-risk (neck size = 41.4 ± 2.8 cm, body mass index = 27.6 ± 3.6) (both P values <.001). An apnea-hypopnea index of ≥5 was found in 2 (8.3%, 95% CI 1.0-20.0%) high-risk and 2 (7.7, 95% CI 1.0-18.4%) low-risk players. Two offensive linemen, a linebacker, and a tight end accounted for the positive cases. CONCLUSIONS: Based on our sample, we estimate the prevalence of SDB among collegiate football players to be 8%, regardless of risk stratification. Given the strong link between SDB and cardiovascular disease, these data underscore the importance of screening and subsequent treatment of SDB in this highly conditioned yet potentially vulnerable group of athletes.

Inter-observer and intra-observer reliability of mechanical axis alignment before and after total knee arthroplasty using long leg radiographs.

BACKGROUND: Long leg radiographs (LLRs) are commonly performed for assessment of mechanical alignment and operative planning in patients undergoing total knee arthroplasty (TKA). The aim of this study was to determine the inter- and intra-observer reliability of alignment measured by observers of different levels of experience. METHODS: Forty patients on the waiting list for a TKA had pre- and post-operative standardised LLRs. We analysed the measurements of mechanical axis alignment between an orthopaedic surgeon, a senior orthopaedic registrar, a junior orthopaedic registrar, and a medical student. Reviewers performed blinded measurements on the same computer screen. These measurements were repeated three months later to assess intra-observer reliability. Furthermore high-resolution screens were compared with standard hospital computer screens to investigate whether monitoring quality influenced the accuracy of measurements of alignment. RESULTS: Inter-observer reliability was high for pre-operative LLRs with an intra-class correlation (ICC) of >0.9 at all experience levels. Post-operative ICC was lowest between the surgeon and the medical student at 0.7. Intra-observer reliability was high at all experience levels. Larger deformities appeared to have exaggerated measurements for both pre- and post-operative images. There appeared to be no effect of the monitor size and quality on the accuracy of measurement. CONCLUSIONS: Long leg radiographs can be used to measure mechanical axis alignment with strong reliability across different levels of experience. This information is important for the evaluation of knee alignment measurements in current clinical practice, to assess severity of deformity and to accompany pre-operative planning and post-operative evaluation. LEVEL OF EVIDENCE: III.

Language ability of children with and without a history of stuttering: a longitudinal cohort study.

PURPOSE: This study aims to determine whether the communication and language skills of children who have a history of stuttering are different from children who do not have a history of stuttering at ages 2-5 years. METHOD: This study utilizes data from the Early Language in Victoria Study (ELVS), a longitudinal study with a community sample of 1910 children recruited in Melbourne, Australia, as well as a concurrent study examining the onset and progression of stuttering. Participants with a history of stuttering (n = 181) and a control group without a history of stuttering (n = 1438) were identified according to the established protocol of these two existing studies. RESULT: The stuttering group scored higher than the non-stuttering group on all of the communication and language outcomes measured. The group differences were statistically significant on four of the seven measures and these findings were maintained when potentially confounding factors were controlled for. CONCLUSION: Importantly, the children with a history of stuttering, as a group, and the control group without a history of stuttering demonstrated developmentally-appropriate early communication and language skills.

Local anesthesia affects physiologic parameters and reduces anesthesiologist intervention in children undergoing general anesthesia for dental rehabilitation.

PURPOSE: The purpose of this study was to evaluate the use of intraoperative local anesthetics in pediatric outpatient dental surgery. This study assessed physiologic stability, as defined by fluctuations in end-tidol carbon dioxide, heart rate, and respiratory rate, as well as subsequent anesthesiologist intervention. METHODS: Forty-eight children (mean age = 3.87 years [+/- l.06 SD]) were included in this randomized, parallel-design study. Intervention variables and the research design were validated in a pilot study. Following collection of baseline vital signs, patients were either given local anesthesia before comprehensive dental treatment or not. Vital sign change was recorded 30 seconds after each procedure. RESULTS: In the no local anesthetic group, 2 areas were found to be statistically significant: (1) postextraction end-tidal carbon dioxide; and (2) heart rate. There was a statistically significant relationship between local anesthetic use and anesthesiologist intervention when assessing the pooled data (P = .001). CONCLUSION: Patients who were not given intraoperative local anesthesia were more likely to experience vital sign fluctuation requiring anesthesiologist intervention.

A Preventive Intervention Program for Urban African American Youth Attending an Alternative Education Program: Background, Implementation, and Feasibility.

This paper presents background, implementation, and feasibility findings associated with planning and conducting an after-school intervention program in an alternative education setting designed to prevent the initiation and escalation of violence and substance abuse among urban African American youth at high risk for life-long problem behaviors. Evolving from earlier preventive interventions implemented in clinic and school settings, the program, entitled The Village Model of Care, consisted of structured group mentoring, parental support, and community outreach services administered to alternative education students and their primary caregiver(s) during the school year. Over a two-year intake period, 109 youth participated in the present process evaluation study. Findings from the study not only provided relevant demographic information on the characteristics of youth likely to be included in such programs but also indicated the importance of including the family in the rehabilitation effort and the need for school administrative system support for the underlying alternative education approach. The information presented in this report has a direct bearing on the planning of future prevention efforts conducted in similar settings that are aimed at reducing problem behaviors and promoting positive lifestyles among high-risk youth.