Role of Nesprin-2 and RanBP2 in BICD2-associated brain developmental disorders.

Bicaudal D2 (BICD2) is responsible for recruiting cytoplasmic dynein to diverse forms of subcellular cargo for their intracellular transport. Mutations in the human BICD2 gene have been found to cause an autosomal dominant form of spinal muscular atrophy (SMA-LED2), and brain developmental defects. Whether and how the latter mutations are related to roles we and others have identified for BICD2 in brain development remains little understood. BICD2 interacts with the nucleoporin RanBP2 to recruit dynein to the nuclear envelope (NE) of Radial Glial Progenitor cells (RGPs) to mediate their well-known but mysterious cell-cycle-regulated interkinetic nuclear migration (INM) behavior, and their subsequent differentiation to form cortical neurons. We more recently found that BICD2 also mediates NE dynein recruitment in migrating post-mitotic neurons, though via a different interactor, Nesprin-2. Here, we report that Nesprin-2 and RanBP2 compete for BICD2-binding in vitro. To test the physiological implications of this behavior, we examined the effects of known BICD2 mutations using in vitro biochemical and in vivo electroporation-mediated brain developmental assays. We find a clear relationship between the ability of BICD2 to bind RanBP2 vs. Nesprin-2 in controlling of nuclear migration and neuronal migration behavior. We propose that mutually exclusive RanBP2-BICD2 vs. Nesprin-2-BICD2 interactions at the NE play successive, critical roles in INM behavior in RGPs and in post-mitotic neuronal migration and errors in these processes contribute to specific human brain malformations.

Risk-based management of international sporting events during the COVID-19 pandemic.

Mass gatherings include a diverse range of events such as sporting competitions, religious ceremonies, entertainment activities, political rallies and cultural celebrations, which have important implications for population well-being. However, if not managed properly, these events can amplify health risks including those related to communicable diseases, and place undue strain on health systems in host countries and potentially in attendees' home countries, upon their return. The coronavirus disease 2019 (COVID-19) pandemic has provided a unique opportunity to evaluate the risk factors associated with mass gatherings and the effectiveness of applying mitigation measures during infectious disease emergencies. The pandemic has also allowed event organizers and health officials to identify best practices for mass gathering planning in host countries. To guide decisions about whether to hold, postpone, modify or cancel a mass gathering during the COVID-19 pandemic, the World Health Organization and its partners developed normative guidance and derivative tools promoting a risk-based approach to mass gathering planning. This approach involves three steps to guide decision-making around mass gatherings: risk evaluation, risk mitigation and risk communication. The approach was applied in the planning and execution of several mass gathering events, including the Tokyo 2020 and Beijing 2022 Olympic and Paralympic Games. Lessons identified from these large-scale international events offer insights into the planning and implementation of mass gathering events during a pandemic, and the broader impacts of such events on society. These lessons may also further inform and refine planning for future mass gatherings.

Les rassemblements de masse désignent un large éventail d'événements tels que des compétitions sportives, cérémonies religieuses, activités de divertissement, manifestations politiques et fêtes culturelles. Tous ont un impact considérable sur le bien-être de la population. Toutefois, s'ils ne sont pas gérés correctement, ils peuvent augmenter les risques sanitaires, notamment concernant les maladies transmissibles, et exercer une pression excessive sur les systèmes de santé des pays hôtes, voire sur ceux des pays d'origine des participants après leur retour. La pandémie de maladie à coronavirus 2019 (COVID-19) a offert une occasion unique d'évaluer les facteurs de risque associés aux rassemblements de masse, ainsi que l'efficacité des mesures visant à limiter la propagation dans des situations d'urgence liées à des maladies infectieuses. Cette pandémie a également permis aux organisateurs d'événements et responsables de santé d'identifier les bonnes pratiques à appliquer dans les pays hôtes pendant les rassemblements de masse. Afin de guider les décisions relatives au maintien, au report, à la modification ou à l'annulation d'un rassemblement de masse durant la pandémie de COVID-19, l'Organisation mondiale de la Santé et ses partenaires ont mis au point des orientations normatives et des outils dérivés favorisant une approche tenant compte des risques au moment de la planification. Cette approche comprend trois étapes contribuant à la prise de décision: l'évaluation, la réduction et la communication des risques. Elle a été déployée lors de la planification et de l'exécution de nombreux rassemblements de masse, comme les Jeux olympiques et paralympiques de Tokyo 2020 et Beijing 2022. Les leçons tirées de ces événements internationaux à grande échelle fournissent des informations sur leur organisation et leur mise en œuvre en cas de pandémie, ainsi que les impacts de tels événements sur la société. Elles sont en outre susceptibles de faciliter et d'améliorer la planification des futurs rassemblements de masse.

Las concentraciones masivas incluyen una gran variedad de eventos, como competiciones deportivas, ceremonias religiosas, actividades de entretenimiento, mítines políticos y celebraciones culturales, que tienen importantes implicaciones para el bienestar de la población. Sin embargo, si no se gestionan adecuadamente, estos eventos pueden amplificar los riesgos para la salud, incluidos los relacionados con las enfermedades transmisibles, y suponer una carga excesiva para los sistemas sanitarios de los países anfitriones y, potencialmente, de los países de origen de los participantes a su regreso. La pandemia de la enfermedad por coronavirus de 2019 (COVID-19) ha brindado una oportunidad única para evaluar los factores de riesgo asociados a las concentraciones masivas y la eficacia de aplicar medidas de mitigación durante las emergencias por enfermedades infecciosas. La pandemia también ha permitido a los organizadores de eventos y a las autoridades sanitarias identificar las mejores prácticas para la planificación de concentraciones masivas en los países anfitriones. Para orientar las decisiones sobre la celebración, el aplazamiento, la modificación o la cancelación de una concentración masiva durante la pandemia de la COVID-19, la Organización Mundial de la Salud y sus asociados elaboraron orientaciones normativas y herramientas derivadas que promueven un enfoque de la planificación de concentraciones masivas basado en los riesgos. Este enfoque consta de tres pasos para orientar la toma de decisiones en torno a las concentraciones masivas: la evaluación, la mitigación y la comunicación de riesgos. El enfoque se aplicó en la planificación y ejecución de varias concentraciones masivas, incluidos los Juegos Olímpicos y Paralímpicos de Tokio 2020 y Pekín 2022. Las conclusiones extraídas de estos eventos internacionales a gran escala permiten comprender mejor la planificación y ejecución de concentraciones masivas durante una pandemia, así como las repercusiones más generales de estos eventos en la sociedad. Estas lecciones también pueden informar y perfeccionar la planificación de futuras concentraciones masivas.

Accelerated atherosclerosis in beta-thalassemia.

Children with beta-thalassemia (BT) present with an increase in carotid intima-medial thickness, an early sign suggestive of premature atherosclerosis. However, it is unknown if there is a direct relationship between BT and atherosclerotic disease. To evaluate this, wild-type (WT, littermates) and BT (Hbbth3/+) mice, both male and female, were placed on a 3-mo high-fat diet with low-density lipoprotein receptor suppression via overexpression of proprotein convertase subtilisin/kexin type 9 (PCSK9) gain-of-function mutation (D377Y). Mechanistically, we hypothesize that heme-mediated oxidative stress creates a proatherogenic environment in BT because BT is a hemolytic anemia that has increased free heme and exhausted hemopexin, heme's endogenous scavenger, in the vasculature. We evaluated the effect of hemopexin (HPX) therapy, mediated via an adeno-associated virus, to the progression of atherosclerosis in BT and a phenylhydrazine-induced model of intravascular hemolysis. In addition, we evaluated the effect of deferiprone (DFP)-mediated iron chelation in the progression of atherosclerosis in BT mice. Aortic en face and aortic root lesion area analysis revealed elevated plaque accumulation in both male and female BT mice compared with WT mice. Hemopexin therapy was able to decrease plaque accumulation in both BT mice and mice on our phenylhydrazine (PHZ)-induced model of hemolysis. DFP decreased atherosclerosis in BT mice but did not provide an additive benefit to HPX therapy. Our data demonstrate for the first time that the underlying pathophysiology of BT leads to accelerated atherosclerosis and shows that heme contributes to atherosclerotic plaque development in BT.NEW & NOTEWORTHY This work definitively shows for the first time that beta-thalassemia leads to accelerated atherosclerosis. We demonstrated that intravascular hemolysis is a prominent feature in beta-thalassemia and the resulting increases in free heme are mechanistically relevant. Adeno-associated virus (AAV)-hemopexin therapy led to decreased free heme and atherosclerotic plaque area in both beta-thalassemia and phenylhydrazine-treated mice. Deferiprone-mediated iron chelation led to deceased plaque accumulation in beta-thalassemia mice but provided no additive benefit to hemopexin therapy.

Rapid implementation of an emergency on-site CKRT dialysate production system during the COVID-19 pandemic.

BACKGROUND: On December 29, 2021, during the delta wave of the Coronavirus Disease 2019 (COVID-19) pandemic, the stock of premanufactured solutions used for continuous kidney replacement therapy (CKRT) at the University of New Mexico Hospital (UNMH) was nearly exhausted with no resupply anticipated due to supply chain disruptions. Within hours, a backup plan, devised and tested 18 months prior, to locally produce CKRT dialysate was implemented. This report describes the emergency implementation and outcomes of this on-site CKRT dialysate production system. METHODS: This is a single-center retrospective case series and narrative report describing and reporting the outcomes of the implementation of an on-site CKRT dialysate production system. All adults treated with locally produced CKRT dialysate in December 2021 and January 2022 at UNMH were included. CKRT dialysate was produced locally using intermittent hemodialysis machines, hemodialysis concentrate, sterile parenteral nutrition bags, and connectors made of 3-D printed biocompatible rigid material. Outcomes analyzed included dialysate testing for composition and microbiologic contamination, CKRT prescription components, patient mortality, sequential organ failure assessment (SOFA) scores, and catheter-associated bloodstream infections (CLABSIs). RESULTS: Over 13 days, 22 patients were treated with 3,645 L of locally produced dialysate with a mean dose of 20.0 mL/kg/h. Fluid sample testing at 48 h revealed appropriate electrolyte composition and endotoxin levels and bacterial colony counts at or below the lower limit of detection. No CLABSIs occurred within 7 days of exposure to locally produced dialysate. In-hospital mortality was 81.8% and 28-day mortality was 68.2%, though illness severity was high, with a mean SOFA score of 14.5. CONCLUSIONS: Though producing CKRT fluid with IHD machines is not novel, this report represents the first description of the rapid and successful implementation of a backup plan for local CKRT dialysate production at a large academic medical center in the U.S. during the COVID-19 pandemic. Though conclusions are limited by the retrospective design and limited sample size of our analysis, our experience could serve as a guide for other centers navigating similar severe supply constraints in the future.

NOXA1-dependent NADPH oxidase 1 signaling mediates angiotensin II activation of the epithelial sodium channel.

The activity of the epithelial Na+ channel (ENaC) in principal cells of the distal nephron fine-tunes renal Na+ excretion. The renin-angiotensin-aldosterone system modulates ENaC activity to control blood pressure, in part, by influencing Na+ excretion. NADPH oxidase activator 1-dependent NADPH oxidase 1 (NOXA1/NOX1) signaling may play a key role in angiotensin II (ANG II)-dependent activation of ENaC. The present study aimed to explore the role of NOXA1/NOX1 signaling in ANG II-dependent activation of ENaC in renal principal cells. Patch-clamp electrophysiology and principal cell-specific Noxa1 knockout (PC-Noxa1 KO) mice were used to determine the role of NOXA1/NOX1 signaling in ANG II-dependent activation of ENaC. The activity of ENaC in the luminal plasma membrane of principal cells was quantified in freshly isolated split-opened tubules using voltage-clamp electrophysiology. ANG II significantly increased ENaC activity. This effect was robust and observed in response to both acute (40 min) and more chronic (48-72 h) ANG II treatment of isolated tubules and mice, respectively. Inhibition of ANG II type 1 receptors with losartan abolished ANG II-dependent stimulation of ENaC. Similarly, treatment with ML171, a specific inhibitor of NOX1, abolished stimulation of ENaC by ANG II. Treatment with ANG II failed to increase ENaC activity in principal cells in tubules isolated from the PC-Noxa1 KO mouse. Tubules from wild-type littermate controls, though, retained their ability to respond to ANG II with an increase in ENaC activity. These results indicate that NOXA1/NOX1 signaling mediates ANG II stimulation of ENaC in renal principal cells. As such, NOXA1/NOX1 signaling in the distal nephron plays a central role in Na+ homeostasis and control of blood pressure, particularly as it relates to regulation by the renin-ANG II axis.NEW & NOTEWORTHY Activity of the epithelial Na+ channel (ENaC) in the distal nephron fine-tunes renal Na+ excretion. Angiotensin II (ANG II) has been reported to enhance ENaC activity. Emerging evidence suggests that NADPH oxidase (NOX) signaling plays an important role in the stimulation of ENaC by ANG II in principal cells. The present findings indicate that NOX activator 1/NOX1 signaling mediates ANG II stimulation of ENaC in renal principal cells.

Evaluating educational interventions to increase breast density awareness among Latinas: A randomized trial in a Federally Qualified Health Center.

BACKGROUND: The objective of this randomized trial was to evaluate the short-term effect of bilingual written and interpersonal education regarding mammographic breast density (MBD). METHODS: Latinas aged 40 to 74 years who were presenting for screening mammography were recruited and randomized 1:1:1 to receive a letter with their mammogram and MBD results (usual care [UC]), a letter plus a brochure (enhanced care [ENH]), or a letter plus a brochure and telephonic promotora education (interpersonal care [INT]). Surveys were administered at enrollment (T0 ) and 2 weeks to 6 months after intervention delivery (T1 ). Differences were assessed with χ2 , Kruskal-Wallis, and McNemar tests and pairwise comparisons as appropriate. INT metrics and audio recordings were analyzed with descriptive statistics and qualitative content analysis. RESULTS: Between October 2016 and October 2019, 943 of 1108 Latina participants (85%) completed both surveys. At T1 , INT participants were more likely (P < .001) to report seeing their MBD results in the letter (70.2%) than UC (53.1%) or ENH participants (55.1%). The percentage of INT women who reported speaking with a provider about MBD (29.0%) was significantly greater (P < .001) than the percentage of UC (14.7%) or ENH participants (15.6%). All groups saw significant (P < .001) but nondifferential improvements in their knowledge of MBD as a masking and risk factor. In the INT group, the promotora delivered education to 77.1% of the 446 participants randomized to INT and answered questions at 28.3% of the encounters for an average of $4.70 per participant. CONCLUSIONS: Among Latinas in a low-resource setting, MBD knowledge may increase with written or interpersonal education, but with modest investment, interpersonal education may better improve MBD awareness and prompt patient-provider discussions.

Butyrate supplementation to pregnant mice elicits cytoprotection against colonic injury in the offspring.

BACKGROUND: Maternal diet during pregnancy can impact progeny health and disease by influencing the offspring's gut microbiome and immune development. Gut microbial metabolism generates butyrate, a short-chain fatty acid that benefits intestinal health. Here we assess the effects of antenatal butyrate on the offspring's gastrointestinal health. We hypothesized that antenatal butyrate supplementation will induce protection against colitis in the offspring. METHODS: C57BL/6 mice received butyrate during pregnancy and a series of experiments were performed on their offspring. RNA sequencing was performed on colonic tissue of 3-week-old offspring. Six-8-week-old offspring were subjected to dextran sulfate sodium-induced colitis. Fecal microbiome analysis was performed on the 6-8-week-old offspring. RESULTS: Antenatal butyrate supplementation dampened transcript enrichment of inflammation-associated colonic genes and prevented colonic injury in the offspring. Antenatal butyrate increased the offspring's stool microbiome diversity and expanded the prevalence of specific gut microbes. CONCLUSIONS: Antenatal butyrate supplementation resulted in downregulation of genes in the offspring's colon that function in inflammatory signaling. In addition, antenatal butyrate supplementation was associated with protection against colitis and an expanded fecal microbiome taxonomic diversity in the offspring. IMPACT: Dietary butyrate supplementation to pregnant mice led to downregulation of colonic genes involved in inflammatory signaling and cholesterol synthesis, changes in the fecal microbiome composition of the offspring, and protection against experimentally induced colitis in the offspring. These data support the mounting evidence that the maternal diet during pregnancy has enduring effects on the offspring's long-term health and disease risk. Although further investigations are needed to identify the mechanism of butyrate's effects on fetal gut development, the current study substantiates the approach of dietary intervention during pregnancy to optimize the long-term gastrointestinal health of the offspring.

Optogenetic Control of PIP2 Interactions Shaping ENaC Activity.

The activity of the epithelial Na+ Channel (ENaC) is strongly dependent on the membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2). PIP2 binds two distinct cationic clusters within the N termini of ß- and γ-ENaC subunits (ßN1 and γN2). The affinities of these sites were previously determined using short synthetic peptides, yet their role in sensitizing ENaC to changes in PIP2 levels in the cellular system is not well established. We addressed this question by comparing the effects of PIP2 depletion and recovery on ENaC channel activity and intracellular Na+ levels [Na+]i. We tested effects on ENaC activity with mutations to the PIP2 binding sites using the optogenetic system CIBN/CRY2-OCRL to selectively deplete PIP2. We monitored changes of [Na+]i by measuring the fluorescent Na+ indicator, CoroNa Green AM, and changes in channel activity by performing patch clamp electrophysiology. Whole cell patch clamp measurements showed a complete lack of response to PIP2 depletion and recovery in ENaC with mutations to ßN1 or γN2 or both sites, compared to wild type ENaC. Whereas mutant ßN1 also had no change in CoroNa Green fluorescence in response to PIP2 depletion, γN2 did have reduced [Na+]i, which was explained by having shorter CoroNa Green uptake and half-life. These results suggest that CoroNa Green measurements should be interpreted with caution. Importantly, the electrophysiology results show that the ßN1 and γN2 sites on ENaC are each necessary to permit maximal ENaC activity in the presence of PIP2.

Eyewitness confidence and mock juror decisions of guilt: A meta-analytic review.

OBJECTIVE: We investigated the impact of eyewitness confidence on the following dependent variables: (a) guilty or not-guilty verdict; (b) judgments of guilt as measured on a scale; and (c) mock jurors' perception of the accuracy of an eyewitness's identification. In addition, we examined two potential moderators of the effects of eyewitness confidence: (a) whether the eyewitness expressed confidence at trial versus during the initial lineup identification and (b) whether the eyewitness provided a numerical versus a verbal statement of confidence. HYPOTHESES: We expected all analyses to reveal that highly confident eyewitnesses are more persuasive to mock jurors than are eyewitnesses with lower confidence (Hypothesis 1). We expected eyewitness confidence at trial (relative to at identification) to be more persuasive to mock jurors (Hypothesis 2). We expected numerical expressions of confidence to be more persuasive to mock jurors than verbal confidence expressions (Hypothesis 3). METHOD: We conducted a meta-analysis of 35 studies from 20 published papers and seven theses or dissertations to quantify the effect of eyewitness confidence on juror judgments and investigated the influence of two primary moderator variables, time of confidence and format of confidence expression. RESULTS: All analyses revealed an effect of eyewitness confidence on mock juror decisions (gs = .21-.36). Our moderator analysis showed that the timing of the confidence statement (identification vs. trial) did not affect the influence of eyewitness confidence on mock jurors' judgments of guilt or accuracy. The influence of eyewitness confidence was not moderated by verbal versus numerical expressions of confidence. CONCLUSIONS: Although eyewitness confidence is persuasive to mock jurors, the size of this effect is modest. Moreover, verbal and numerical expressions of confidence have similar persuasive effects, and mock jurors do not appear to be sensitive to the likely difference in evidentiary strength of eyewitness confidence expressed at the initial identification versus at trial. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Phosphatidylinositol 4,5-bisphosphate directly interacts with the ß and γ subunits of the sodium channel ENaC.

The plasma membrane phospholipid phosphatidylinositol 4,5-bisphosphate (PIP2) regulates the activity of diverse ion channels to include the epithelial Na+ channel ENaC. Whether PIP2 regulation of ENaC is due to a direct phospholipid-protein interaction, remains obscure. To date, possible interaction of PIP2 with ENaC primarily has been tested indirectly through assays of channel function. A fragment-based biochemical analysis approach is used here to directly quantify possible PIP2-ENaC interactions. We find using the CIBN-CRY2 optogenetic dimerization system that the phosphoryl group positioned at carbon 5 of PIP2 is necessary for interaction with ENaC. Previous studies have implicated conserved basic residues in the cytosolic portions of ß- and γ-ENaC subunits as being important for PIP2-ENaC interactions. To test this, we used synthetic peptides of these regions of ß- and γ-ENaC. Steady-state intrinsic fluorescence spectroscopy demonstrated that phosphoinositides change the local conformation of the N terminus of ß-ENaC, and two sites of γ-ENaC adjacent to the plasma membrane, suggesting direct interactions of PIP2 with these three regions. Microscale thermophoresis elaborated PIP2 interactions with the N termini of ß- (Kd ∼5.2 µm) and γ-ENaC (Kd ∼13 µm). A weaker interaction site within the carboxyl terminus of γ-ENaC (Kd ∼800 µm) was also observed. These results support that PIP2 regulates ENaC activity by directly interacting with at least three distinct regions within the cytoplasmic domains of the channel that contain conserved basic residues. These interactions are probably electrostatic in nature, and are likely to bear a key structural role in support of channel activity.

Lactococcus lactis Subspecies cremoris Elicits Protection Against Metabolic Changes Induced by a Western-Style Diet.

BACKGROUND & AIMS: A Western-style diet, which is high in fat and sugar, can cause significant dyslipidemia and nonalcoholic fatty liver disease; the diet has an especially strong effect in women, regardless of total calorie intake. Dietary supplementation with beneficial microbes might reduce the detrimental effects of a Western-style diet. We assessed the effects of Lactococcus lactis subspecies (subsp) cremoris on weight gain, liver fat, serum cholesterol, and insulin resistance in female mice on a high-fat, high-carbohydrate diet. METHODS: Female C57BL/6 mice were fed either a high-fat, high-carbohydrate (Western-style) diet that contained 40% fat (mostly milk fat) and 43% carbohydrate (mostly sucrose) or a calorie-matched-per-gram control diet. The diets of mice were supplemented with 1 × 109 colony-forming units of L lactis subsp cremoris ATCC 19257 or Lactobacillus rhamnosus GG ATCC 53103 (control bacteria) 3 times per week for 16 weeks. Body weights were measured, and fecal, blood, and liver tissues were collected and analyzed. Livers were analyzed for fat accumulation and inflammation, and blood samples were analyzed for cholesterol and glucose levels. Mice were housed within Comprehensive Lab Animal Monitoring System cages, and respiratory exchange ratio and activity were measured. Hepatic lipid profiles of L lactis subsp cremoris-supplemented mice were characterized by lipidomics mass spectrometry analysis. RESULTS: Mice fed L lactis subsp cremoris while on the Western-style diet gained less weight, developed less hepatic steatosis and inflammation, and had a lower mean serum level of cholesterol and body mass index than mice fed the control bacteria. Mice fed the L lactis subsp cremoris had increased glucose tolerance while on the Western-style diet compared to mice fed control bacteria and had alterations in hepatic lipids, including oxylipins. CONCLUSIONS: Dietary supplementation with L lactis subsp cremoris in female mice on a high-fat, high-carbohydrate (Western-style) diet caused them to gain less weight, develop less liver fat and inflammation, reduce serum cholesterol levels, and increase glucose tolerance compared with mice on the same diet fed control bacteria. L lactis subsp cremoris is safe for oral ingestion and might be developed for persons with metabolic and liver disorders caused by a Western-style diet.

Automated Digital Notification of COVID-19 Diagnoses Through Text and Email Messaging - North Carolina, December 2020-January 2021.

During October 3, 2020-January 9, 2021, North Carolina experienced a 400% increase in daily reported COVID-19 cases (1). To handle the increased number of cases and rapidly notify persons receiving a positive SARS-CoV-2 test result (patients), North Carolina state and local health departments moved from telephone call notification only to telephone call plus automated text and email notification (digital notification) beginning on December 24, 2020. Overall, among 200,258 patients, 142,975 (71%) were notified by telephone call or digital notification within the actionable period (10 days from their diagnosis date)* during January 2021, including at least 112,543 (56%) notified within 24 hours of report to North Carolina state and local health departments, a significantly higher proportion than the 25,905 of 175,979 (15%) notified within 24 hours during the preceding month (p<0.001). Differences in text notification by age, race, and ethnicity were observed. Automated digital notification is a feasible, rapid and efficient method to support timely outreach to patients, provide guidance on how to isolate, access resources, inform close contacts, and increase the efficiency of case investigation staff members.

Promoting water consumption among children: a three-arm cluster randomised controlled trial testing a social network intervention.

OBJECTIVE: To test the effectiveness of a social network intervention (SNI) to improve children's healthy drinking behaviours. DESIGN: A three-arm cluster randomised control trial design was used. In the SNI, a subset of children were selected and trained as 'influence agents' to promote water consumption-as an alternative to sugar-sweetened beverages (SSB)-among their peers. In the active control condition, all children were simultaneously exposed to the benefits of water consumption. The control condition received no intervention. SETTING: Eleven schools in the Netherlands. PARTICIPANTS: Four hundred and fifty-one children (Mage = 10·74, SDage = 0·97; 50·8 % girls). RESULTS: Structural path models showed that children exposed to the SNI consumed 0·20 less SSB per day compared to those in the control condition (ß = 0·25, P = 0·035). There was a trend showing that children exposed to the SNI consumed 0·17 less SSB per day than those in the active control condition (ß = 0·20, P = 0·061). No differences were found between conditions for water consumption. However, the moderation effects of descriptive norms (ß = -0·12, P = 0·028) and injunctive norms (ß = 0·11-0·14, both P = 0·050) indicated that norms are more strongly linked to water consumption in the SNI condition compared to the active control and control conditions. CONCLUSIONS: These findings suggest that a SNI promoting healthy drinking behaviours may prevent children from consuming more SSB. Moreover, for water consumption, the prevailing social norms in the context play an important role in mitigating the effectiveness of the SNI.

Promoting water consumption among Dutch children: an evaluation of the social network intervention Share H2O.

BACKGROUND: There is a need to develop and improve interventions promoting healthy drinking behaviors among children. A promising method could be to stimulate peer influence within children's social networks. In the Share H2O social network intervention (SNI), peer influence was utilized by selecting a subset of influential children and training them as 'influence agents' to promote water consumption-as an alternative to SSBs. Previous research has mainly focused on the process of selecting influence agents. However, the process of motivating influence agents to promote the behavior has hardly received any research attention. Therefore, in the SNI Share H2O SNI, this motivation process was emphasized and grounded in the self-determination theory (SDT). This study evaluated the implementation of the Share H2O SNI, focusing on whether and how applying SDT-based techniques can motivate the influence agents and, indirectly, their peers. METHODS: This study included data collected in the Netherlands from both the influence agents (n = 37) and the peers (n = 112) in the classroom networks of the influence agents. Self-reported measurements assessed the influence agents' enjoyment of the training, duration and perceived autonomy support during the training, and changes in their intrinsic motivation and water consumption before and after the start of the intervention. Changes in the peers' intrinsic motivation, perceived social support, and social norms were measured before and after the start of the intervention. RESULTS: The influence agents enjoyed the training, the duration was adequate, and perceived it as autonomy supportive. There was an increase in the influence agents' intrinsic motivation to drink water and their actual water consumption. Providing personal meaningful rationales seemed to have motivated the influence agents. The intrinsic motivation and perceived descriptive norm of the peers remained stable. The peers reported an increase in their perceived social support and injunctive norm concerning water drinking after the intervention. Influence agents appeared to mainly use face-to-face strategies, such as modeling, talking to peers, and providing social support to promote the behavior. CONCLUSIONS: The current findings provided preliminary evidence of the promising effects of using SDT-based techniques in an SNI to motivate the influence agents and, indirectly, their peers. TRIAL REGISTRATION: NTR, NL6905, Registered 9 January 2018, https://www.trialregister.nl/trial/6905.

Evaluation of perfusion index as a noninvasive tool to determine epidural anesthesia effectiveness in dogs.

OBJECTIVE: To evaluate perfusion index (PI) as a noninvasive tool to determine effectiveness and onset of epidural anesthesia in dogs. STUDY DESIGN: Prospective clinical trial. ANIMALS: A total of 21 adult dogs, aged 6.5 ± 3 years and weighing 34.9 ± 6.4 kg, undergoing a tibial plateau leveling osteotomy. METHODS: Dogs were premedicated intramuscularly with acepromazine (0.03 mg kg-1) and hydromorphone (0.1 mg kg-1) and anesthetized with intravenous propofol (to effect) and isoflurane in oxygen. A surface transflectance probe was secured to the tail base to monitor PI and a dorsal pedal artery catheter was placed for invasive blood pressure monitoring. A lumbosacral epidural was performed with the dog in sternal recumbency. Dogs were randomly assigned for inclusion of epidural morphine (0.1 mg kg-1) or morphine (0.1 mg kg-1) and lidocaine (4 mg kg-1). PI was recorded following instrumentation of each dog just prior to the epidural (baseline), at 10 minute intervals for 30 minutes, before and after the surgical skin incision and before and after completion of the osteotomy. Physiological variables and end-tidal isoflurane were recorded at the same time points. RESULTS: There was no significant difference in PI between the groups at any time point. There was a significant change in end-tidal isoflurane before and after the skin incision in the epidural morphine and epidural morphine-lidocaine groups (p = 0.04, p = 0.05, respectively) and before and after the osteotomy in each group for heart rate (p = 0.001, p = 0.04), diastolic (p = 0.01, p = 0.01) and mean arterial blood pressure (p = 0.03, p = 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: PI did not provide an objective means for determining the onset or effectiveness of epidural anesthesia in anesthetized dogs and alternate methods of noninvasive assessment should be investigated.

Assessment of Microleakage under Stainless Steel Orthodontic Brackets Bonded with Various Adhesive Systems: An In Vitro Study.

AIM AND OBJECTIVE: The aim of this study was to evaluate the extent of microleakage beneath stainless steel orthodontic brackets bonded with different adhesive systems. MATERIALS AND METHODS: Freshly extracted 60 human premolar teeth from mandibular arch were included in this study. After sterilizing all teeth, they were stored in thymol solution of 1% for further preparation. Acrylic blocks were used to mount the teeth in a way their roots were totally implanted up to the cement enamel junction in acrylic with crown being visible. A 0.022 slot, stainless steel preadjusted edgewise premolar brackets were taken. Sixty premolars were categorized randomly into three groups (20 premolars in each group) as follows: group I: flowable composite, group II: Fuji Ortho LC, group III: Transbond XT. Later, all the samples were subjected to thermocycling and tested immediately and 24 hours after water storage. The samples were submerged for 24 hours in methylene blue solution (2%) at room temperature. A ×20 magnification stereomicroscope was used to examine all samples. RESULTS: The lowest microleakage (1.34 ± 0.20) was shown by Transbond XT restored teeth, followed by flowable composite group (1.79 ± 0.32) and Fuji Ortho LC group (2.98 ± 0.13). An analysis of variance showed statistically significant differences among various adhesive systems. A statistically significant difference (p <0.05) among groups I and II, and groups II and III adhesive materials was seen. CONCLUSION: This study demonstrated microleakage in all the examined adhesive groups but the lowest microleakage was found with Transbond XT group followed next by Filtek Z350 XT group and Fuji Ortho LC group. CLINICAL SIGNIFICANCE: Due to microleakage, the bacteria and fluids present intraorally penetrate through the gaps along the enamel-adhesive boundary. This penetration results in significant esthetic and clinical complications. Such problems related to microleakage can be addressed with the use of an appropriate adhesive agent.

A mutually induced conformational fit underlies Ca2+-directed interactions between calmodulin and the proximal C terminus of KCNQ4 K+ channels.

Calmodulin (CaM) conveys intracellular Ca2+ signals to KCNQ (Kv7, "M-type") K+ channels and many other ion channels. Whether this "calmodulation" involves a dramatic structural rearrangement or only slight perturbations of the CaM/KCNQ complex is as yet unclear. A consensus structural model of conformational shifts occurring between low nanomolar and physiologically high intracellular [Ca2+] is still under debate. Here, we used various techniques of biophysical chemical analyses to investigate the interactions between CaM and synthetic peptides corresponding to the A and B domains of the KCNQ4 subtype. We found that in the absence of CaM, the peptides are disordered, whereas Ca2+/CaM imposed helical structure on both KCNQ A and B domains. Isothermal titration calorimetry revealed that Ca2+/CaM has higher affinity for the B domain than for the A domain of KCNQ2-4 and much higher affinity for the B domain when prebound with the A domain. X-ray crystallography confirmed that these discrete peptides spontaneously form a complex with Ca2+/CaM, similar to previous reports of CaM binding KCNQ-AB domains that are linked together. Microscale thermophoresis and heteronuclear single-quantum coherence NMR spectroscopy indicated the C-lobe of Ca2+-free CaM to interact with the KCNQ4 B domain (Kd ∼10-20 µm), with increasing Ca2+ molar ratios shifting the CaM-B domain interactions via only the CaM C-lobe to also include the N-lobe. Our findings suggest that in response to increased Ca2+, CaM undergoes lobe switching that imposes a dramatic mutually induced conformational fit to both the proximal C terminus of KCNQ4 channels and CaM, likely underlying Ca2+-dependent regulation of KCNQ gating.

Neutrophil-Derived Reactive Oxygen Orchestrates Epithelial Cell Signaling Events during Intestinal Repair.

Recent evidence has demonstrated that reactive oxygen (eg, hydrogen peroxide) can activate host cell signaling pathways that function in repair. We show that mice deficient in their capacity to generate reactive oxygen by the NADPH oxidase 2 holoenzyme, an enzyme complex highly expressed in neutrophils and macrophages, have disrupted capacity to orchestrate signaling events that function in mucosal repair. Similar observations were made for mice after neutrophil depletion, pinpointing this cell type as the source of the reactive oxygen driving oxidation-reduction protein signaling in the epithelium. To simulate epithelial exposure to high levels of reactive oxygen produced by neutrophils and gain new insight into this oxidation-reduction signaling, epithelial cells were treated with hydrogen peroxide, biochemical experiments were conducted, and a proteome-wide screen was performed using isotope-coded affinity tags to detect proteins oxidized after exposure. This analysis implicated signaling pathways regulating focal adhesions, cell junctions, and maintenance of the cytoskeleton. These pathways are also known to act via coordinated phosphorylation events within proteins that constitute the focal adhesion complex, including focal adhesion kinase and Crk-associated substrate. We identified the Rho family small GTP-binding protein Ras-related C3 botulinum toxin substrate 1 and p21 activated kinases 2 as operational in these signaling and localization pathways. These data support the hypothesis that reactive oxygen species from neutrophils can orchestrate epithelial cell-signaling events functioning in intestinal repair.

Successful use of once-daily high-dose darunavir and dolutegravir in multidrug-resistant HIV.

WHAT IS KNOWN AND OBJECTIVE?: Antiretroviral (ARV) resistance may result during periods of consistently poor adherence. We report the successful use of a novel once-daily (QD) ARV regimen in a patient with multidrug-resistant (MDR) HIV. CASE SUMMARY: Once-daily darunavir 1200 mg/ritonavir 100 mg, dolutegravir and emtricitabine/tenofovir alafenamide was initiated with directly observed therapy. With the assistance of therapeutic drug monitoring, dolutegravir dosing was increased to 150 mg daily. The patient maintained virologic suppression for 18 months. WHAT IS NEW AND CONCLUSIONS?: In this case, QD darunavir/ritonavir achieved similar trough concentrations to twice daily dosing with dolutegravir dose titration necessitated and resulted in HIV virologic control.

Comparison of 3 Surveillance Methods to Detect Potential Controlled Substance Diversion in an Academic Medical Center.

Objectives: To compare 3 methods of detecting potential diversion of controlled substances (CS) by health care personnel from inpatient units in a large, academic medical center. Methods: Three different reports were retrospectively analyzed and evaluated to determine which employees are "high-risk" for diversion over a 30-day period using defined criteria. Reports were derived from automated dispensing machines (ADMs), purchased third-party software (TPS), and the electronic health record (EHR). The primary outcome was the percentage of employees in each report who were deemed to be high-risk for CS diversion (positive predictive value [PPV]). Secondary outcomes included the number of false positives and description of high-risk users on each report. Descriptive statistics were used to analyze differences between methods. Results: The PPV was highly variable between reports. The PPVs among the ADM, TPS, and EHR reports were 3.28%, 6.82%, and 23.88%, respectively. False positives were high among all reports (96.72%, 93.18%, and 76.12% for the ADM, TPS, and EHR reports, respectively). Conclusions: A report from the EHR has the highest PPV to detect high-risk employees who may be diverting CS. However, false positives were high for all reports, indicating that significant improvements are needed in the development of accurate and reliable software to detect potential and actual CS diversion.