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Ligand field excitation of hexaamminerhodium(III), Rh(NH3)63+, in aqueous solution results in the photoaquation of one coordinated NH3. The measured quantum yields at 23° are 0.075 mol/einstein for 313-nm excitation of the 1A1g → 1T1g transition and 0.07 for 254-nm excitation of the 1A1g → 1T2g transition, suggesting efficient interconversion of the higher energy state to a common reactive excited state. Under similar conditions, the quantum yield for the perdeuterated complex Rh(ND3)63+ is about twice as large (0.14 in H2O, 0.15 in D2O, 313-nm excitation). However, the temperature dependence of the photoaquation quantum yields is measurably greater for the perprotio complex than for the perdeuterio complex. To explain these results, it is proposed that temperature-independent weak coupling and temperature-dependent strong coupling mechanisms are competitive in the nonradiative deactivation of the reactive excited state of Rh(NH3)63+.

Mitochondrial dysfunction causes devastating disorders, including mitochondrial myopathy, but how muscle senses and adapts to mitochondrial dysfunction is not well understood. Here, we used diverse mouse models of mitochondrial myopathy to show that the signal for mitochondrial dysfunction originates within mitochondria. The mitochondrial proteins OMA1 and DELE1 sensed disruption of the inner mitochondrial membrane and, in response, activated the mitochondrial integrated stress response (mt-ISR) to increase the building blocks for protein synthesis. In the absence of the mt-ISR, protein synthesis in muscle was dysregulated causing protein misfolding, and mice with early-onset mitochondrial myopathy failed to grow and survive. The mt-ISR was similar following disruptions in mtDNA maintenance (Tfam knockout) and mitochondrial protein misfolding (CHCHD10 G58R and S59L knockin) but heterogenous among mitochondria-rich tissues, with broad gene expression changes observed in heart and skeletal muscle and limited changes observed in liver and brown adipose tissue. Taken together, our findings identify that the DELE1 mt-ISR mediates a similar response to diverse forms of mitochondrial stress and is critical for maintaining growth and survival in early-onset mitochondrial myopathy., Competing Interests: Disclosure and competing interests statement The authors declare no competing interests., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Objective: Anorexia nervosa (AN) is associated with increased risk of mortality, but little is known about the risk of inpatient admissions and mortality outcomes in individuals with diagnoses of both AN and other psychiatric and somatic conditions. We aimed to investigate the inpatient admissions and mortality among people with AN and other diagnosed conditions using Danish national registers., Method: This retrospective cohort study included individuals diagnosed with AN in Denmark, born 1977-2010. We identified other mental and somatic conditions in this population. We used Cox proportional hazards regression to estimate the risk of inpatient admission and mortality, focusing on (i) the number of other diagnosed conditions, and (ii) specific combinations of conditions diagnosed prior to the AN diagnosis. Categories of inpatient admissions considered were due to: (i) AN, (ii) any psychiatric disorder, and (iii) any somatic disorder. Additionally, competing risks survival analysis was used to calculate the cumulative incidence of inpatient admission and all-cause mortality over the follow-up period., Results: The study population included 11,489 individuals. The most common conditions individuals had prior to their AN diagnosis were other eating disorders (34.5%) and anxiety disorders (32.7%). During the follow-up, 3184 (27.7%), 4604 (40.1%), and 6636 (57.8%) individuals were admitted for AN, any psychiatric disorder, and any somatic disorder, respectively; and in total 106 (0.9%) died. The risk of all outcomes was highest among those who had received a higher number of other diagnoses. For most combinations, the risks of admission and mortality were increased., Discussion: Our study presents the prevalence of other conditions in patients with AN in Denmark and elucidates their association with higher rates of inpatient admission and mortality. Our findings highlight the need for comprehensive, multidisciplinary care of patients with AN considering the spectrum of other diagnosed conditions to improve health outcomes., (© 2024 The Authors. Acta Psychiatrica Scandinavica published by John Wiley & Sons Ltd.)

Background: Ulcerative colitis is a chronic inflammatory disease of the colon with an unknown etiology. Alkaline sphingomyelinase (alk-SMase) is specifically expressed by intestinal epithelial cells, and has been reported to play an anti-inflammatory role. However, the underlying mechanism is still unclear., Aim: To explore the mechanism of alk-SMase anti-inflammatory effects on intestinal barrier function and oxidative stress in dextran sulfate sodium (DSS)-induced colitis., Methods: Mice were administered 3% DSS drinking water, and disease activity index was determined to evaluate the status of colitis. Intestinal permeability was evaluated by gavage administration of fluorescein isothiocyanate dextran, and bacterial translocation was evaluated by measuring serum lipopolysaccharide. Intestinal epithelial cell ultrastructure was observed by electron microscopy. Western blotting and quantitative real-time reverse transcription-polymerase chain reaction were used to detect the expression of intestinal barrier proteins and mRNA, respectively. Serum oxidant and antioxidant marker levels were analyzed using commercial kits to assess oxidative stress levels., Results: Compared to wild-type (WT) mice, inflammation and intestinal permeability in alk-SMase knockout (KO) mice were more severe beginning 4 d after DSS induction. The mRNA and protein levels of intestinal barrier proteins, including zonula occludens-1, occludin, claudin-3, claudin-5, claudin-8, mucin 2, and secretory immunoglobulin A, were significantly reduced on 4 d after DSS treatment. Ultrastructural observations revealed progressive damage to the tight junctions of intestinal epithelial cells. Furthermore, by day 4, mitochondria appeared swollen and degenerated. Additionally, compared to WT mice, serum malondialdehyde levels in KO mice were higher, and the antioxidant capacity was significantly lower. The expression of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) in the colonic mucosal tissue of KO mice was significantly decreased after DSS treatment. mRNA levels of Nrf2-regulated downstream antioxidant enzymes were also decreased. Finally, colitis in KO mice could be effectively relieved by the injection of tertiary butylhydroquinone, which is an Nrf2 activator., Conclusion: Alk-SMase regulates the stability of the intestinal mucosal barrier and enhances antioxidant activity through the Nrf2 signaling pathway., Competing Interests: Conflict-of-interest statement: The authors report no relevant conflicts of interest for this article., (©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.)

Background: HIV-1 CRF65_cpx strain carries drug-resistant mutations, which raises concerns about its potential for causing virologic failure. The CRF65_cpx ranks as the fourth most prevalent on Hainan Island, China. However, the origin and molecular epidemiology of CRF65_cpx strains in this area remain unclear. This study aims to estimate the spatial origins and dissemination patterns of HIV-1 CRF65_cpx in this specific region., Methods: Between 2018 and 2021, a total of 58 pol sequences of the CRF65_cpx were collected from HIV-positive patients on Hainan Island. The available CRF65_cpx pol sequences from public databases were compiled. The HIV-TRACE tool was used to construct transmission networks. The evolutionary history of the introduction and dissemination of HIV-1 CRF65_cpx on Hainan Island were analyzed using phylogenetic analysis and the Bayesian coalescent-based approach., Results: Among the 58 participants, 89.66% were men who have sex with men (MSM). The median age was 25 years, and 43.10% of the individuals had a college degree or above. The results indicated that 39 (67.24%) sequences were interconnected within a single transmission network. A consistent expansion was evident from 2019 to 2021, with an incremental annual addition of four sequences into the networks. Phylodynamic analyses showed that the CRF65_cpx on Hainan Island originated from Beijing (Bayes factor, BF = 17.4), with transmission among MSM on Hainan Island in 2013.2 (95%HPD: 2012.4, 2019.5), subsequently leading to an outbreak. Haikou was the local center of the CRF65_cpx epidemic. This strain propagated from Haikou to other locations, including Sanya (BF > 1000), Danzhou (BF = 299.3), Chengmai (BF = 27.0) and Tunchang (BF = 16.3). The analyses of the viral migration patterns between age subgroups and risk subgroups revealed that the viral migration directions were from "25-40 years old" to "17-24 years old" (BF = 14.6) and to "over 40 years old" (BF = 17.6), and from MSM to heterosexuals (BF > 1000) on Hainan Island., Conclusion: Our analyses elucidate the transmission dynamics of CRF65_cpx strain on Hainan Island. Haikou is identified as the potential hotspot for CRF65_cpx transmission, with middle-aged MSM identified as the key population. These findings suggest that targeted interventions in hotspots and key populations may be more effective in controlling the HIV epidemic., (© 2024. The Author(s).)

Mitochondrial dysfunction causes devastating disorders, including mitochondrial myopathy. Here, we identified that diverse mitochondrial myopathy models elicit a protective mitochondrial integrated stress response (mt-ISR), mediated by OMA1-DELE1 signaling. The response was similar following disruptions in mtDNA maintenance, from knockout of Tfam , and mitochondrial protein unfolding, from disease-causing mutations in CHCHD10 (G58R and S59L). The preponderance of the response was directed at upregulating pathways for aminoacyl-tRNA biosynthesis, the intermediates for protein synthesis, and was similar in heart and skeletal muscle but more limited in brown adipose challenged with cold stress. Strikingly, models with early DELE1 mt-ISR activation failed to grow and survive to adulthood in the absence of Dele1 , accounting for some but not all of OMA1's protection. Notably, the DELE1 mt-ISR did not slow net protein synthesis in stressed striated muscle, but instead prevented loss of translation-associated proteostasis in muscle fibers. Together our findings identify that the DELE1 mt-ISR mediates a stereotyped response to diverse forms of mitochondrial stress and is particularly critical for maintaining growth and survival in early-onset mitochondrial myopathy., Competing Interests: Conflict-of-interest statement The authors declare that they have no conflict of interest.

The receptor tyrosine kinase (RTK) KIT and its ligand stem cell factor (SCF) are essential for human mast cell (huMC) survival and proliferation. HuMCs expressing oncogenic KIT variants secrete large numbers of extracellular vesicles (EVs). The role KIT plays in regulating EV secretion has not been examined. Here, we investigated the effects of stimulation or inhibition of KIT activity on the secretion of small EVs (sEVs). In huMCs expressing constitutively active KIT, the quantity and quality of secreted sEVs positively correlated with the activity status of KIT. SCF-mediated stimulation of KIT in huMCs or murine MCs, or of transiently expressed KIT in HeLa cells, enhanced the release of sEVs expressing exosome markers. In contrast, ligand-mediated stimulation of the RTK EGFR in HeLa cells did not affect sEV secretion. The release of sEVs induced by either constitutively active or ligand-activated KIT was remarkably decreased when cells were treated with KIT inhibitors, concomitant with reduced exosome markers in sEVs. Similarly, inhibition of oncogenic KIT signalling kinases like PI3K, and MAPK significantly reduced the secretion of sEVs. Thus, activation of KIT and its early signalling cascades stimulate the secretion of exosome-like sEVs in a regulated fashion, which may have implications for KIT-driven functions., Competing Interests: The authors report no conflict of interest., (© 2024 The Authors. Journal of Extracellular Biology published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)

Objectives: Whether the routine delivery of diabetes-related knowledge can change patients' attitudes and hence influence their self-management activities remains unknown in primary healthcare settings in China. Thus, this study aims to explore the complex transformation process between knowledge, attitude and practice (KAP) among patients with diabetes in a city in China., Design: A cross-sectional study., Setting: Yuhuan City, Zhejiang Province, China., Participants: A total of 803 patients with diabetes were invited to attend a questionnaire survey and 782 patients with type 2 diabetes completed the survey. The average age of participants was 58.47 years old, 48.21% of whom only attended primary school or below., Primary and Secondary Outcome Measures: A questionnaire based on existing scales and expert consultation was applied to assess patients' socio-demographic information (SI), disease progression risk and diabetes-related KAP. A structural equation model was built to analyse the relationships between patients' characteristics and KAP., Results: No significant association was found between patients' knowledge and attitude (β=0.01, p=0.43). Better knowledge and attitude were both found to be associated with better diet and physical activities (β=0.58, p<0.001; β=0.46, p=0.01). However, patients with a more positive attitude toward diabetic care showed worse foot care practice (β=-0.13, p=0.02), while better knowledge was associated with better foot care practice (β=0.29, p<0.001). In addition, patients with higher SI (β=0.88, p<0.001) and/or disease progression risk (β=0.42, p<0.001) tended to present higher levels of disease knowledge., Conclusions: While successful KAP transformation has been achieved in practice for diet and physical activities, there is a need to improve foot care practice. Health education should also prioritise the prevention, detection and care of diabetic foot. Also, appropriate methods should be adopted to deliver health education to vulnerable patients, such as the elderly, those living in rural areas, those with minimal education, the unemployed and low-income patients., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Food sociality refers to the exploration of food production, exchange, distribution, and consumption and influences on cultural communication and social meaning. This study aimed to investigate food sociality in three provinces of South China to provide theoretical and practical evidence of food sociality in the region and to revise nutrition policies., Materials and Methods: We conducted a qualitative study comprising 25 experts in the fields of nutrition, sociology, food science, and agriculture from Hainan, Guangdong, and Guangxi Province by using a semi-structured in-depth interview, which included 28 pre-determined questions covering six topics. The interviews were conducted between November 2020 and March 2021. Verbatim transcripts were analyzed thematically using NVivo 11.0., Results: Of the 25 experts, the mean age was 50.6 (SD = 8.4) years, 15 (60%) were male, and 22 (88%) held a master's degree or above. The analysis showed that food sociality in three provinces of South China mainly comprises social functions of food and dietary behavior. Regarding social functions of food, the experts expressed that food represents local culture (72%, 18 experts), presents social status and economic power (40%, 10 experts), and is central to special occasions, traditional customs, and etiquette activities (60%, 15 experts). In terms of social functions of dietary behaviors, the majority of experts indicated that food is a social communication tool (72% experts), has geographical characteristics (80% experts), and, to some extent, is used as a proxy for reward or punishment. Furthermore, festivals are one of the core elements of food sociality in the region, although food safety is a major concern. Some dietary behaviors, such as overindulgence in afternoon tea and encouraging drinking, may increase the risk of chronic diseases., Conclusion: Food sociality in three provinces of South China is mainly related to the social function of food and dietary behavior. It is a combination of local culture, social status and economic power, traditional customs, rewards and punishments, geographical food preference, and social communication tools. The authors recommend increasing food safety at festivals and promoting healthy eating behaviors in order to improve the overall health of the population in this region., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Yan, Petersen, Lin, Chen, Li, Ji and Zhang.)

Introduction: In the US, opioids are commonly prescribed after arthroscopic knee surgery. We aimed to investigate opioid dispensing in relation to arthroscopic knee surgeries from 2006 to 2018 in Denmark., Methods: In Danish registries, we identified patients (≥ 15 years old) having arthroscopic knee surgery (anterior cruciate ligament (ACL) reconstruction; meniscal surgery; cartilage resection; synovectomy and diagnostic arthroscopy) between 1 January 2006 and 31 December 2018 and opioid dispensing (oral morphine equivalents (OMEQ)) within seven days after discharge from surgery., Results: Among 218,940 patients, 15,263 (7%) had an opioid dispensed within seven days after being discharged following surgery. The opioid dispensing incidence (per 1,000 persons/year) increased during the study period for all procedures. This trend was more pronounced for ACL reconstruction, which recorded an increase from 86 (95% confidence interval (CI): 75-99) in 2006 to 278 (95% CI: 255-301) in 2018, corresponding to 9% and 28% of ACL patients, respectively. In the same period, the average amount of opioids dispensed within the first seven days after discharge decreased (change: 70.0 OMEQ (95% CI: 12.4-127.5)). Tramadol and oxycodone were the most commonly dispensed opioids., Conclusion: About 7% of patients had an opioid dispensed within the first seven days after discharge following arthroscopic knee surgery. The incidence of opioids dispensed increased for all investigated procedures from 2006 to 2018. In the same period, the average amount of opioids dispensed within the first seven days after discharge decreased., Funding: None., Trial Registration: Not relevant., (Articles published in the DMJ are “open access”. This means that the articles are distributed under the terms of the Creative Commons Attribution Non-commercial License, which permits any non-commercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.)

Research studies and conservation actions aimed at improving conditions for bees require a basic understanding of which species are present in a given region. The US state of Minnesota occupies a unique geographic position at the confluence of eastern deciduous forests, northern boreal forests, and western tallgrass prairie, which has led to a diverse and unique bee fauna. In recent years there have been multiple ongoing bee-focused inventory and research projects in Minnesota. Combined with the historic specimens housed in the University of Minnesota Insect Collection and other regional collections, these furnished a wealth of specimens available to form the basis of a statewide checklist. Here, we present the first comprehensive checklist of Minnesota bee species, documenting a total of 508 species in 45 genera. County-level occurrence data is included for each species, and further information on distribution and rarity is included for species of regional or national interest. Some species have their taxonomy clarified, with Perdita citrinella Graenicher, 1910 syn. nov. recognized as a junior synonym of Perdita perpallida Cockerell, 1901, P. bequaerti syn. nov. recognized as a junior synonym of P. pallidipennis Graenicher, 1910 stat. nov., Anthidiellum boreale (Robertson, 1902) stat. nov. recognized as a full species, and Anthidiellium beijingense Portman & Ascher nom. nov. is proposed for A. boreale Wu to resolve the homonymy with A. boreale (Robertson). We further include a list of species that may occur in Minnesota and highlight 11 species occurring in the state that are considered non-native. Recent collecting efforts, as well as increased taxonomic attention paid to Minnesota bees, have resulted in 66 species that have only been documented in the last 10 years. As a first step in determining native bees of conservation concern, we document 38 species that have not been detected in the state during the last 50 years and discuss their conservation status, along with other species for which evidence of decline exists. The checklist of Minnesota bees will continue to grow and change with additional surveys and research studies. In particular, recent surveys have continued to detect new bee species, and many bee groups are in need of taxonomic revision, with the most recent revisions for many genera occurring decades ago. Overall, this checklist strengthens our understanding of the bees of Minnesota and the broader region, informs conservation assessments, and establishes a baseline for faunal change.

Environmental pollution seriously affects human health. The concentration of negative air ions (NAIs), which were discovered at the end of the nineteenth century, is one of the factors used to evaluate air quality. Additionally, NAIs have been widely considered markers by scholars due to their unique biological function. The aim of this study was to summarize existing research and propose future research on the generation and temporal and spatial dynamic patterns of NAIs concentrations as well as the relationship between NAIs and human health. We identified 187 studies (published January 2013-January 2023) that met our inclusion criteria. Fourteen English studies evaluated the effects of NAIs on depression, the cardiovascular system, the respiratory system, reproduction and development, cognition, and sports muscle injury. Only two studies reported the associations of NAIs exposure with metabolic omics. NAIs concentrations vary temporally with solar radiation, air temperature, and relative humidity, while the temporal dynamic patterns of NAIs are affected by season, time, meteorological factors, air quality index, geographical location, forest vegetation, and other factors. Researchers have shown that exposure to NAIs may benefit our health by changing amino acid metabolism, which mainly manifests as increased anti-inflammation and reduced inflammation and antioxidation. Furthermore, exposure to NAIs promotes energy production, affects the expression of c-fos, and regulates 5-HT levels. There has been considerable interest in the potential effects of NAIs on human health and well-being, but the conclusions have been inconsistent and the mechanisms remain unclear. The use of omics to elucidate the biological mechanism of NAIs is relatively new and has some advantages., (© 2023. The Author(s).)

Extracellular vesicles (EVs) released by endothelial cells support vascular homeostasis. To better understand endothelial cell EV biogenesis, we examined cultured human umbilical vein endothelial cells (HUVECs) prepared by rapid freezing, freeze-substitution, and serial thin section electron microscopy (EM). Thin sections of HUVECs revealed clusters of membrane protrusions on the otherwise smooth cell surface. The protrusions contained membrane-bound organelles, including multivesicular bodies (MVBs), and appeared to be on the verge of pinching off to form microvesicles. Beyond cell peripheries, membrane-bound vesicles with internal MVBs were observed, and serial sections confirmed that they were not connected to cells. These observations are consistent with the notion that these multi-compartmented microvesicles (MCMVs) pinch-off from protrusions. Remarkably, omega figures formed by fusion of vesicles with the MCMV limiting membrane were directly observed, apparently releasing exosomes from the MCMV. In summary, MCMVs are a novel form of EV that bud from membrane protrusions on the HUVEC surface, contain MVBs and release exosomes. These observations suggest that exosomes can be harboured within and released from transiting microvesicles after departure from the parent cell, constituting a new site of exosome biogenesis occurring from endothelial and potentially additional cell types., Competing Interests: The authors declare no conflict of interest., (© 2023 The Authors. Journal of Extracellular Biology published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)

Background: Though critical to primary care, continuity of care has rarely been examined in China. This study aims to assess the relationship between continuity of care and healthcare costs among patients with chronic diseases within primary care settings in China., Methods: In this cross-sectional study, we used a social health insurance claims dataset of 1406 patients with hypertension and/or diabetes in Yuhuan City, Zhejiang Province collected in 2017-2019. We measured continuity of care using the Bice-Boxerman Continuity of Care (COC) Index, Herfindahl Index (HI), Sequential Continuity of Care (SECON) Index, Usual Provider of Care (UPC), and a binary variable indicating whether a patient's UPC was a primary care provider. We examined the associations between continuity of care and healthcare costs in the same period and the subsequent year, using ordinary least squares regression for the outpatient costs and two-part regression for the inpatient costs. Based on the regression coefficients, we predicted costs saved if each continuity measure was set to 1 from the status quo., Results: When optimum continuity were to be achieved, 7.12-27.29% of total outpatient costs and 55.38-73.35% of total inpatient costs could be saved compared to the status quo during the two-year study period. If optimum continuity were to be achieved in the first year, 7.47%-21.78% of total outpatient costs and 8.84-40.22% of total inpatient costs could be saved in the second-year., Conclusions: Care continuity indicators were consistently associated with reduced outpatient costs and hospitalization risks. Future health reform in China should further enhance continuity of care in primary care., Competing Interests: The authors have no competing interests to declare., (Copyright: © 2022 The Author(s).)

Activating mutations in the receptor KIT promote the dysregulated proliferation of human mast cells (huMCs). The resulting neoplastic huMCs secrete extracellular vesicles (EVs) that can transfer oncogenic KIT among other cargo into recipient cells. Despite potential contributions to diseases, KIT-containing EVs have not been thoroughly investigated. Here, we isolated and characterized KIT-EV subpopulations released by neoplastic huMCs using an immunocapture approach that selectively isolates EVs containing KIT in its proper topology. Immunocapture of EVs on KIT antibody-coated electron microscopy (EM) affinity grids allowed to assess the morphology and size of KIT-EVs. Immunoblot analysis demonstrated KIT-EVs have a distinct protein profile from KIT-depleted EVs, contain exosome and microvesicle markers, and are separated into these subtypes by ultracentrifugation. Cell treatment with sphingomyelinase inhibitors shifted the protein content among KIT-EV subtypes, suggesting different biogenesis routes. Proteomic analysis revealed huMC KIT-EVs are enriched in proteins involved in signalling, immune responses, and cell migration, suggesting diverse biological functions, and indicated neoplastic huMCs disseminate KIT via shuttling in heterogeneous microvesicle- and exosome-like EVs. Further, selective KIT-immunocapture will enable the enrichment of specific huMC-derived EVs from complex human biosamples and facilitate an understanding of their in vivo functions and potential to serve as biomarkers of specific biological pathologies., (© 2022 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)

CD47 is a marker of self and a signaling receptor for thrombospondin-1 that is also a component of extracellular vesicles (EVs) released by various cell types. Previous studies identified CD47-dependent functional effects of T cell EVs on target cells, mediated by delivery of their RNA contents, and enrichment of specific subsets of coding and noncoding RNAs in CD47 + EVs. Mass spectrometry was employed here to identify potential mechanisms by which CD47 regulates the trafficking of specific RNAs to EVs. Specific interactions of CD47 and its cytoplasmic adapter ubiquilin-1 with components of the exportin-1/Ran nuclear export complex were identified and confirmed by coimmunoprecipitation. Exportin-1 is known to regulate nuclear to cytoplasmic trafficking of 5'-7-methylguanosine (m 7 G)-modified microRNAs and mRNAs that interact with its cargo protein EIF4E. Interaction with CD47 was inhibited following alkylation of exportin-1 at Cys 528 by its covalent inhibitor leptomycin B. Leptomycin B increased levels of m 7 G-modified RNAs, and their association with exportin-1 in EVs released from wild type but not CD47-deficient cells. In addition to perturbing nuclear to cytoplasmic transport, transcriptomic analyses of EVs released by wild type and CD47-deficient Jurkat T cells revealed a global CD47-dependent enrichment of m 7 G-modified microRNAs and mRNAs in EVs released by CD47-deficient cells. Correspondingly, decreasing CD47 expression in wild type cells or treatment with thrombospondin-1 enhanced levels of specific m 7 G-modified RNAs released in EVs, and re-expressing CD47 in CD47-deficient T cells decreased their levels. Therefore, CD47 signaling limits the trafficking of m 7 G-modified RNAs to EVs through physical interactions with the exportin-1/Ran transport complex., (© 2021. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Individuals infected with the SARS-CoV-2 Delta variant, lineage B.1.617.2, exhibit faster initial infection with a higher viral load than prior variants, and pseudotyped viral particles bearing the SARS-CoV-2 Delta variant spike protein induce a faster initial infection rate of target cells compared to those bearing other SARS-CoV-2 variant spikes. Here, we show that pseudotyped viral particles bearing the Delta variant spike form unique aggregates, as evidenced by negative stain and cryogenic electron microscopy (EM), flow cytometry, and nanoparticle tracking analysis. Viral particles pseudotyped with other SARS-CoV-2 spike variants do not show aggregation by any of these criteria. The contribution to infection kinetics of the Delta spike's unique property to aggregate is discussed with respect to recent evidence for collective infection by other viruses. Irrespective of this intriguing possibility, spike-dependent aggregation is a new functional parameter of spike-expressing viral particles to evaluate in future spike protein variants.

Individuals infected with the SARS-CoV-2 Delta variant, lineage B.1.617.2, exhibit faster initial infection with a higher viral load than prior variants, and pseudotyped particles bearing the SARS-CoV-2 Delta variant spike protein induce a faster initial infection rate of target cells compared to those bearing other SARS-CoV-2 variant spikes. Here, we show that pseudotyped particles bearing the Delta variant spike form unique aggregates, as evidenced by negative stain and cryogenic electron microscopy (EM), flow cytometry, and nanoparticle tracking analysis. Viral particles pseudotyped with other SARS-CoV-2 spike variants do not show aggregation by any of these criteria. The contribution to infection kinetics of the Delta spike’s unique property to aggregate is discussed with respect to recent evidence for collective infection by other viruses. Irrespective of this intriguing possibility, spike-dependent aggregation is a new functional parameter of spike-expressing viral particles to evaluate in future spike protein variants.

Neurosurgical clipping and endovascular coiling are both standard therapies to prevent rebleeding after aneurysmal subarachnoid hemorrhage (aSAH). However, controversy still exists about which is the optimal treatment. This meta-analysis aims to assess the effectiveness and safety of two treatments with high-quality evidence. Web of Science, Cochrane Library, EMBASE, Pubmed, Sinomed, China National Knowledge Infrastructure, and Wanfang Data databases were systematically searched on August 5, 2021. Randomized controlled trials (RCTs) and prospective cohort studies that evaluated the effectiveness and safety of clipping versus coiling in aSAH patients at discharge or within 1-year follow-up period were eligible. No restriction was set on the publication date. Meta-analyses were conducted to calculate the pooled estimates and 95% confidence intervals (CI) of relative risk (RR). Eight RCTs and 20 prospective cohort studies were identified. Compared to coiling, clipping was associated with a lower rebleeding rate at discharge (RR: 0.52, 95% CI: 0.29--0.94) and a higher aneurysmal occlusion rate (RR: 1.33, 95% CI: 1.19-1.48) at 1-year follow-up. In contrast, coiling reduced the vasospasm rate at discharge (RR: 1.45, 95% CI: 1.23-1.71) and 1-year poor outcome rate (RR: 1.27, 95% CI: 1.16-1.39). Subgroup analyses presented that among patients with a poor neurological condition at admission, no statistically significant outcome difference existed between the two treatments. The overall prognosis was better among patients who received coiling, but this advantage was not significant among patients with a poor neurological condition at admission. Therefore, the selection of treatment modality for aSAH patients should be considered comprehensively., (© 2021. The Author(s).)

Effective small molecule therapies to combat the SARS-CoV-2 infection are still lacking as the COVID-19 pandemic continues globally. High throughput screening assays are needed for lead discovery and optimization of small molecule SARS-CoV-2 inhibitors. In this work, we have applied viral pseudotyping to establish a cell-based SARS-CoV-2 entry assay. Here, the pseudotyped particles (PP) contain SARS-CoV-2 spike in a membrane enveloping both the murine leukemia virus (MLV) gag-pol polyprotein and luciferase reporter RNA. Upon addition of PP to HEK293-ACE2 cells, the SARS-CoV-2 spike protein binds to the ACE2 receptor on the cell surface, resulting in priming by host proteases to trigger endocytosis of these particles, and membrane fusion between the particle envelope and the cell membrane. The internalized luciferase reporter gene is then expressed in cells, resulting in a luminescent readout as a surrogate for spike-mediated entry into cells. This SARS-CoV-2 PP entry assay can be executed in a biosafety level 2 containment lab for high throughput screening. From a collection of 5,158 approved drugs and drug candidates, our screening efforts identified 7 active compounds that inhibited the SARS-CoV-2-S PP entry. Of these seven, six compounds were active against live replicating SARS-CoV-2 virus in a cytopathic effect assay. Our results demonstrated the utility of this assay in the discovery and development of SARS-CoV-2 entry inhibitors as well as the mechanistic study of anti-SARS-CoV-2 compounds. Additionally, particles pseudotyped with spike proteins from SARS-CoV-2 B.1.1.7 and B.1.351 variants were prepared and used to evaluate the therapeutic effects of viral entry inhibitors., Competing Interests: Declaration of Competing Interests The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2022. Published by Elsevier Inc.)

Seventy five percent of the world's food crops benefit from insect pollination. Hence, there has been increased interest in how global change drivers impact this critical ecosystem service. Because standardized data on crop pollination are rarely available, we are limited in our capacity to understand the variation in pollination benefits to crop yield, as well as to anticipate changes in this service, develop predictions, and inform management actions. Here, we present CropPol, a dynamic, open, and global database on crop pollination. It contains measurements recorded from 202 crop studies, covering 3,394 field observations, 2,552 yield measurements (i.e., berry mass, number of fruits, and fruit density [kg/ha], among others), and 47,752 insect records from 48 commercial crops distributed around the globe. CropPol comprises 32 of the 87 leading global crops and commodities that are pollinator dependent. Malus domestica is the most represented crop (32 studies), followed by Brassica napus (22 studies), Vaccinium corymbosum (13 studies), and Citrullus lanatus (12 studies). The most abundant pollinator guilds recorded are honey bees (34.22% counts), bumblebees (19.19%), flies other than Syrphidae and Bombyliidae (13.18%), other wild bees (13.13%), beetles (10.97%), Syrphidae (4.87%), and Bombyliidae (0.05%). Locations comprise 34 countries distributed among Europe (76 studies), North America (60), Latin America and the Caribbean (29), Asia (20), Oceania (10), and Africa (7). Sampling spans three decades and is concentrated on 2001-2005 (21 studies), 2006-2010 (40), 2011-2015 (88), and 2016-2020 (50). This is the most comprehensive open global data set on measurements of crop flower visitors, crop pollinators and pollination to date, and we encourage researchers to add more datasets to this database in the future. This data set is released for non-commercial use only. Credits should be given to this paper (i.e., proper citation), and the products generated with this database should be shared under the same license terms (CC BY-NC-SA)., (© 2021 The Ecological Society of America.)

Physical activity and obesity are known to be associated. We investigated whether a change in leisure time physical activities (LTPA) predicts a subsequent weight change, or vice versa.We used data from a longitudinal study among Danish adults surveyed in 1983-1984, 1987-1988, and 1993-1994. Between two sequential surveys, the change in LTPA was grouped as no change, became less or more active; the change in body weight was defined as no change, lost or gained of more than one body mass index (BMI) unit.Among 2386 adults, change in LTPA was not associated with subsequent weight change. However, a loss in body weight (BMI change < -1 unit) was associated with subsequent either becoming less [OR = 1.49, 95% CI (1.03-2.15)] or borderline more active [OR = 1.37, 95% CI (0.99-1.90)]. Subgroup analyses showed particularity among females that a loss in body weight was associated with subsequent becoming more active [OR = 1.83, 95% CI (1.15-2.89)].Our results suggest that change in LTPA is unrelated to subsequent weight change, but loss in body weight seems related to subsequent more active among female adults.

T cells and endothelial cells engage in bidirectional communication that regulates angiogenesis and T cell transmigration. Extracellular vesicles (EVs) mediate intercellular communication by the transfer of bioactive molecules including RNAs. EVs produced by a given cell type are heterogeneous in their RNA content, but it is unclear how specific EV surface markers relate to their functional effects on target cells. Our previous work established that Jurkat T cell EVs bearing CD63, MHC-I, or CD47 surface markers contain distinct noncoding RNA populations. The present study reveals that CD63 + and MHC-I + EVs from CD47-deficient Jurkat T cells are enriched in small non-coding RNAs relative to EVs from wild-type Jurkat T cells. CD47-deficient Jurkat T cells secrete more CD63 + and MHC-I + EVs, but MHC-I + EVs are selectively taken up more by human umbilical vein endothelial cells. Transcriptomics analysis of endothelial cells treated with CD63 + or MHC-I + EVs showed surface marker- and CD47-dependent changes in gene expression in the target cells. Gene set enrichment analysis identified CD47-dependent, and surface marker-dependent effects of T cell EVs on VEGF and inflammatory signaling, cell cycle, and lipid and cholesterol metabolism. Thus, subsets of T cell EVs differentially regulate endothelial cell metabolism and inflammatory and angiogenic responses.

Effective small molecule therapies to combat the SARS-CoV-2 infection are still lacking as the COVID-19 pandemic continues globally. High throughput screening assays are needed for lead discovery and optimization of small molecule SARS-CoV-2 inhibitors. In this work, we have applied viral pseudotyping to establish a cell-based SARS-CoV-2 entry assay. Here, the pseudotyped particles (PP) contain SARS-CoV-2 spike in a membrane enveloping both the murine leukemia virus (MLV) gag-pol polyprotein and luciferase reporter RNA. Upon addition of PP to HEK293-ACE2 cells, the SARS-CoV-2 spike protein binds to the ACE2 receptor on the cell surface, resulting in priming by host proteases to trigger endocytosis of these particles, and membrane fusion between the particle envelope and the cell membrane. The internalized luciferase reporter gene is then expressed in cells, resulting in a luminescent readout as a surrogate for spike-mediated entry into cells. This SARS-CoV-2 PP entry assay can be executed in a biosafety level 2 containment lab for high throughput screening. From a collection of 5,158 approved drugs and drug candidates, our screening efforts identified 7 active compounds that inhibited the SARS-CoV-2-S PP entry. Of these seven, six compounds were active against live replicating SARS-CoV-2 virus in a cytopathic effect assay. Our results demonstrated the utility of this assay in the discovery and development of SARS-CoV-2 entry inhibitors as well as the mechanistic study of anti-SARS-CoV-2 compounds. Additionally, particles pseudotyped with spike proteins from SARS-CoV-2 B.1.1.7 and B.1.351 variants were prepared and used to evaluate the therapeutic effects of viral entry inhibitors.

The endosomal recycling system dynamically tunes synaptic strength, which underlies synaptic plasticity. Exocytosis is involved in the expression of long-term potentiation (LTP), as postsynaptic cleavage of the SNARE (soluble NSF-attachment protein receptor) protein VAMP2 by tetanus toxin blocks LTP. Moreover, induction of LTP increases the exocytosis of transferrin receptors (TfRs) and markers of recycling endosomes (REs), as well as post-synaptic AMPA type receptors (AMPARs). However, the interplay between AMPAR and TfR exocytosis remains unclear. Here, we identify VAMP4 as the vesicular SNARE that mediates most dendritic RE exocytosis. In contrast, VAMP2 plays a minor role in RE exocytosis. LTP induction increases the exocytosis of both VAMP2- and VAMP4-labeled organelles. Knock down (KD) of VAMP4 decreases TfR recycling but increases AMPAR recycling. Moreover, VAMP4 KD increases AMPAR-mediated synaptic transmission, which consequently occludes LTP expression. The opposing changes in AMPAR and TfR recycling upon VAMP4 KD reveal their sorting into separate endosomal populations., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Viral fusion is a critical step in the entry pathway of enveloped viruses and remains a viable target for antiviral exploration. The current approaches for studying fusion mechanisms include ensemble fusion assays, high-resolution cryo-TEM, and single-molecule fluorescence-based methods. While these methods have provided invaluable insights into the dynamic events underlying fusion processes, they come with their own limitations. These often include extensive data and image analysis in addition to experimental time and technical requirements. This work proposes the use of the spin-spin T2 relaxation technique as a sensitive bioanalytical method for the rapid quantification of interactions between viral fusion proteins and lipids in real time. In this study, new liposome-coated iron oxide nanosensors (LIONs), which mimic as magnetic-labeled host membranes, are reported to detect minute interactions occurring between the membrane and influenza's fusion glycoprotein, hemagglutinin (HA). The influenza fusion protein's interaction with the LION membrane is detected by measuring changes in the sensitive spin-spin T2 magnetic relaxation time using a bench-top NMR instrument. More data is gleaned from including the fluorescent dye DiI into the LION membrane. In addition, the effects of environmental factors on protein-lipid interaction that affect fusion such as pH, time of incubation, trypsin, and cholesterol were also examined. Furthermore, the efficacy and sensitivity of the spin-spin T2 relaxation assay in quantifying similar protein/lipid interactions with more native configurations of HA were demonstrated using virus-like particles (VLPs). Shorter domains derived from HA were used to start a reductionist path to identify the parts of HA responsible for the NMR changes observed. Finally, the known fusion inhibitor Arbidol was employed in our spin-spin T2 relaxation-based fusion assay to demonstrate the application of LIONs in real-time monitoring of this aspect of fusion for evaluation of potential fusion inhibitors.

Importance: Low socioeconomic status (SES) has been identified as a risk factor for the development of dementia. However, few studies have focused on the association between SES and dementia diagnostic evaluation on a population level., Objective: To investigate whether household income (HHI) is associated with dementia diagnosis and cognitive severity at the time of diagnosis., Design, Setting, and Participants: This population- and register-based cross-sectional study analyzed health, social, and economic data obtained from various Danish national registers. The study population comprised individuals who received a first-time referral for a diagnostic evaluation for dementia to the secondary health care sector of Denmark between January 1, 2017, and December 17, 2018. Dementia-related health data were retrieved from the Danish Quality Database for Dementia. Data analysis was conducted from October 2019 to December 2020., Exposures: Annual HHI (used as a proxy for SES) for 2015 and 2016 was obtained from Statistics Denmark and categorized into upper, middle, and lower tertiles within 5-year interval age groups., Main Outcomes and Measures: Dementia diagnoses (Alzheimer disease, vascular dementia, mixed dementia, dementia with Lewy bodies, Parkinson disease dementia, or other) and cognitive stages at diagnosis (cognitively intact; mild cognitive impairment but not dementia; or mild, moderate, or severe dementia) were retrieved from the database. Univariable and multivariable logistic and linear regressions adjusted for age group, sex, region of residence, household type, period (2017 and 2018), medication type, and medical conditions were analyzed for a possible association between HHI and receipt of dementia diagnosis., Results: Among the 10 191 individuals (mean [SD] age, 75 [10] years; 5476 women [53.7%]) included in the study, 8844 (86.8%) were diagnosed with dementia. Individuals with HHI in the upper tertile compared with those with lower-tertile HHI were less likely to receive a dementia diagnosis after referral (odds ratio, 0.65; 95% CI, 0.55-0.78) and, if diagnosed with dementia, had less severe cognitive stage (β, -0.16; 95% CI, -0.21 to -0.10). Individuals with middle-tertile HHI did not significantly differ from those with lower-tertile HHI in terms of dementia diagnosis (odds ratio, 0.92; 95% CI, 0.77-1.09) and cognitive stage at diagnosis (β, 0.01; 95% CI, -0.04 to 0.06)., Conclusions and Relevance: The results of this study revealed a social inequality in dementia diagnostic evaluation: in Denmark, people with higher income seem to receive an earlier diagnosis. Public health strategies should target people with lower SES for earlier dementia detection and intervention.

Plasmodium falciparum VAR2CSA binds to chondroitin sulfate A (CSA) on the surface of the syncytiotrophoblast during placental malaria. This interaction facilitates placental sequestration of malaria parasites resulting in severe health outcomes for both the mother and her offspring. Furthermore, CSA is presented by diverse cancer cells and specific targeting of cells by VAR2CSA may become a viable approach for cancer treatment. In the present study, we determined the cryo-electron microscopy structures of the full-length ectodomain of VAR2CSA from P. falciparum strain NF54 in complex with CSA, and VAR2CSA from a second P. falciparum strain FCR3. The architecture of VAR2CSA is composed of a stable core flanked by a flexible arm. CSA traverses the core domain by binding within two channels and CSA binding does not induce major conformational changes in VAR2CSA. The CSA-binding elements are conserved across VAR2CSA variants and are flanked by polymorphic segments, suggesting immune selection outside the CSA-binding sites. This work provides paths for developing interventions against placental malaria and cancer.

Objectives: Patients with dementia have an increased 30-day mortality after hip fracture. We investigated clinical management including time to surgery, out-of-hours admission and surgery, surgery on weekends, surgery volume per ward, and anesthesia technique for this excess mortality risk., Method: This register- and population-based study comprised 12,309 older adults (age 70+) admitted to hospital for a first-time hip fracture in 2013-2014, of whom 11,318 underwent hip fracture surgery. Cox proportional hazards regression models were applied for the analysis., Results: The overall postoperative 30-day mortality was 11.4%. Patients with dementia had a 1.5 times increased mortality risk than those without (HR = 1.50 [95% CI 1.31-1.72]). We observed no time-to-surgery difference by patient dementia status; additionally, the excess mortality risk in patients with dementia could not be explained by the clinical management factors we examined., Conclusions: Increased mortality in patients with dementia could not be explained by the measured preoperative clinical management. Suboptimal handling of postoperative complication and rehabilitation are to be investigated for their role in the witnessed increased mortality for patients with dementia., (© 2020 The Authors. Brain and Behavior published by Wiley Periodicals LLC.)

While vaccine development will hopefully quell the global pandemic of COVID-19 caused by SARS-CoV-2, small molecule drugs that can effectively control SARS-CoV-2 infection are urgently needed. Here, inhibitors of spike (S) mediated cell entry were identified in a high throughput screen of an approved drugs library with SARS-S and MERS-S pseudotyped particle entry assays. We discovered six compounds (cepharanthine, abemaciclib, osimertinib, trimipramine, colforsin, and ingenol) to be broad spectrum inhibitors for spike-mediated entry. This work could contribute to the development of effective treatments against the initial stage of viral infection and provide mechanistic information that might aid the design of new drug combinations for clinical trials for COVID-19 patients., Competing Interests: The authors declare no competing financial interest., (© 2020 American Chemical Society.)

Night shift work suppresses excretion of melatonin, but little is known about the needed time for recovery. We aimed to compare levels of 6-sulfatoxy melatonin after three different night shift schedules, including recovery days. In a quasi-experimental, within-subject crossover study, 73 male police officers in Denmark collected morning urine after the last recovery day in three different work schedules with two, four, and seven consecutive night shifts followed by a corresponding number of days for recovery. We found no significant differences for 6-sulphatoxymelatonin concentrations in morning urine between the three different work schedules indicating similar recovery of melatonin suppression in the studied work schedules.

Background: Persons with dementia tend to be vulnerable to mobility challenges and hence face a greater risk of fall and subsequent fractures, morbidity, and mortality. Motion-based technologies (MBTs), also called sensor-based technologies or virtual reality, have the potential for assisting physical exercise and training as a part of a disease management and rehabilitation program, but little is known about its' use for people with dementia., Objective: The purpose of this pilot study was to investigate the feasibility and efficacy of MBT physical training at home for people with dementia., Methods: A 3-phase pilot study: (1) baseline start-up, (2) 15 weeks of group training at a local care center twice a week, and (3) 12 weeks of group training reduced to once a week, supplemented with individual MBT training twice a week at home. A total of 26 people with dementia from a municipality in Southern Denmark were eligible and agreed to participate in this study. Three withdrew from the study, leaving 23 participants for the final analysis. Feasibility was measured by the percentage of participants who trained with MBT at home, and their completion rate of total scheduled MBT sessions. Efficacy was evaluated by physical function, measured by Sit-to-Stand (STS), Timed-Up-and-Go (TUG), 6-minute Walk Test (6MW), and 10-meter Dual-task Walking Test (10MDW); cognitive function was measured by Mini-Mental State Examination (MMSE) and Neuropsychiatric Inventory-Questionnaire (NPI-Q); and European Quality of Life 5 dimensions questionnaire (EQOL5) was used for measuring quality of life. Descriptive statistics were applied accordingly. Wilcoxon signed-rank and rank-sum tests were applied to explore significant differences within and between the groups., Results: As much as 12 of 23 participants (52%) used the supplemental MBT training at home. Among them, 6 (50%) completed 75% or more scheduled sessions, 3 completed 25% or less, and 3 completed between 25% and 75% of scheduled sessions. For physical and cognitive function tests, supplementing with MBT training at home showed a tendency of overall stabilization of scores among the group of participants who actively trained with MBT; especially, the 10MDW test even showed a significant improvement from 9.2 to 7.1 seconds (P=.03). We found no positive effect on EQOL5 tests., Conclusions: More than half of the study population with dementia used MBT training at home, and among them, half had an overall high adherence to the home training activity. Physical function tended to remain stable or even improved among high-adherence MBT individuals. We conclude that MBT training at home may be feasible for some individuals with dementia. Further research is warranted., (©Jindong Ding Petersen, Eva Ladekjær Larsen, Karen la Cour, Cecilie von Bülow, Malene Skouboe, Jeanette Reffstrup Christensen, Frans Boch Waldorff. Originally published in JMIR Mental Health (http://mental.jmir.org), 26.08.2020.)

Background: Physiologic neonatal hyperbilirubinemia (jaundice) is common and affects most newborn infants. However, there is a risk for permanent neurological damage if the bilirubin levels rise above a certain threshold. The management of neonatal jaundice includes the assessment of bilirubin laboratory values, consideration of patient-specific risk factors, and plotting on a bilirubin nomogram reference to determine risk and guide therapy. When performed manually, the process can be time consuming and error-prone. Therefore, web-based calculators such as BiliTool have been developed to assist in risk assessment., Methods: To streamline the risk assessment calculation process further within our electronic health record (EHR), we created a "BiliReport" to display patient bilirubin-related data and automate transmission of deidentified patient data to the BiliTool website (https://bilitool.org). After implementation, we evaluated usage data, provider satisfaction, and accuracy of documentation., Results: We demonstrated high provider use of the BiliReport and satisfaction with the workflow. We found a significant improvement in the accuracy of bilirubin risk level documentation, with a reduction in erroneous risk stratification from 4% (15/232) to 0.4% (1/243), P < 0.001. We did not find significant a difference in erroneous documentation of the bilirubin lab value (P = 0.07)., Conclusions: Integrating the neonatal hyperbilirubinemia risk assessment process into the EHR may reduce errors and improve provider documentation and adherence to recommended guidelines., (Copyright © 2020 Academic Pediatric Association. Published by Elsevier Inc. All rights reserved.)

Plants may benefit from limiting the community of generalist floral visitors if the species that remain are more effective pollinators and less effective pollenivores. Plants can reduce access to pollen through altered floral cues or morphological structures, but can also reduce consumption through direct pollen defenses. We observed that Eucera (Peponapis) pruinosa, a specialist bee on Cucurbita plants, collected pure loads of pollen while generalist honey bees and bumble bees collected negligible amounts of cucurbit pollen, even though all groups of bees visited these flowers. Cucurbit flowers have no morphological adaptations to limit pollen collection by bees, thus we assessed their potential for physical, nutritional, and chemical pollen traits that might act as defenses to limit pollen loss to generalist pollinators. Bumble bee (Bombus impatiens) microcolonies experienced reduced pollen consumption, mortality, and reproduction as well as increased stress responses when exposed to nutritional and mechanical pollen defenses. These bees also experienced physiological effects of these defenses in the form of hindgut expansion and gut melanization. Chemical defenses alone increased the area of gut melanization in larger bees and induced possible compensatory feeding. Together, these results suggest that generalist bumble bees avoid collecting cucurbit pollen due to the physiological costs of physical and chemical pollen defenses.

Infected T cells and macrophages are the main producers of HIV-1 in infected individuals. Upon release from infected cells, HIV-1 incorporates various cellular membrane proteins, some of which are specific for these cells. However, the functions of cell-encoded proteins in virions remain largely unknown. We performed flow virometry to identify, in plasma of HIV-infected individuals, macrophage- and T-cell-derived HIV-1 virions, using cell-specific markers CD36 and CD27, respectively. Using four different methods, we demonstrated that CD36 on virions binds the immunosuppressive cytokine transforming growth factor beta (TGF-β) through a ligand, thrombospondin one (TSP-1). Flow virometry of individual virions showed that TGF-β was present on CD36+ virions (average, 28.2% ± 6.6% (n = 3)) but not on CD27+ virions (average, 1% ± 0.1% (n = 3)). TGF-β molecules present on captured CD36+ virions were biologically active, as evaluated with a reporter cell line. Delivery of TGF-β on HIV virions to HIV target cells may affect them, playing a significant role in viral pathogenesis.

Aim: We used register-based data to estimate the effect of all-type dementia on road traffic accidents (RTAs) risk, combined with comorbidities or sedative medicines, among non-institutionalized older people in Denmark., Methods: The source population was all residents in Denmark aged 65 years and older, alive as of January 1, 2008 ( n = 853,228). Cases were those who had any type of RTA in 2009-2014. Each case was matched for age, sex and geographic location to 4-6 controls. All-type dementia was ascertained using the International Classification of Diseases version 10 (ICD-10) diagnosis supplemented with prescribed medicine records. Eight chronic diseases were selected to assess comorbidities. Four types of medicines were categorized as sedative medicines for analysis. Conditional logistic regression with adjustment for education and marital status as well as either the number of comorbidities or sedative medications use was performed using STATA software., Results: Older people with dementia had lower RTAs risk compared to their controls (odds ratio = 0.43, 95% confidence interval (0.32-0.60), p < 0.001). Significant interaction was observed between dementia and the number of comorbidities for RTAs estimation., Conclusions: The significantly lower RTAs risk for older individuals with dementia observed in our study may be due to people with dementia living at home having a lower frequency of outdoor activities; that is, less exposure to traffic. However, this, together with the interaction between dementia and comorbidities as well as sedative medications, should be investigated further.

Regulation of AMPA receptor (AMPAR) trafficking is a key modulator of excitatory synaptic transmission; however, intracellular vesicular transport of newly synthesized AMPARs has been little studied due to technical limitations. By combining molecular tools with imaging strategies in cultured rat hippocampal neurons, we found that vesicles containing newly synthesized, GluA1-subunit-containing AMPARs are transported antero- and retrogradely at a mean speed of 1.5 μm.s -1 . Synaptic activity and variations in intracellular calcium levels bidirectionally modulate GluA1 transport. Chemical long-term potentiation (cLTP) initially induces a halt in GluA1 transport, followed by a sustained increase, while acute glutamate uncaging on synaptic spines arrests vesicular movements. GluA1 phosphomimetic mutants preferentially travel to the dendritic tip, probably to replenish extrasynaptic pools, distal to the soma. Our findings indicate that AMPAR intracellular transport is highly regulated during synaptic plasticity and likely controls AMPAR numbers at the plasma membrane., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)

The aim was to describe the organization of working hours in the Danish regions according to sex, age and calendar year. Based on the Danish Working Hour Database (DWHD), individuals were classified according to schedules: Permanent day (57.8%), evening (1.7%), or night (1.2%); day/evening (22.0%); day/night (6.6%); evening/night (0.6%); and day/evening/night (10.2%). More men (9.1%) than women (5.9%) worked day/night, whereas more women (10.9%) than men (7.4%) worked day/evening/night. More young than older employees worked day/evening/night, and fewer worked permanent day or night. From 2008 to 2015 we observed a trend towards more employees working permanent day and fewer employees working other schedules. Altogether DWHD provides a strong tool in research on working hours.

Introduction: Institutionalized people with dementia have an increased risk of fall accidents, but little is known about whether this increased risk holds for home dwellers., Methods: This register- and population-based study comprised 115,584 cases and 394,679 controls. Cases were individuals with any fall between 2009 and 2014, and matched with up to six controls on age, sex, and geographic location. Individuals were excluded if they (1) had any fall in 2008, or (2) lived in a nursing home on the date of the fall. Dementia, other chronic diseases, and sedative medicines were assessed from Danish national registers., Results: After adjusting for potential confounders, older people with dementia living at home had a 1.89-fold higher risk of fall (odds ratio = 1.89, 95% confidence interval [1.84-1.94], P  < .001)., Discussion: Dementia almost doubles the risk of fall for older Danish people living at home. This highlights the need for effective fall preventions that target people with dementia.

Long-term potentiation (LTP) of excitatory synaptic transmission has long been considered a cellular correlate for learning and memory. Early LTP (less than 1 h) had initially been explained either by presynaptic increases in glutamate release or by direct modification of postsynaptic AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptor function. Compelling models have more recently proposed that synaptic potentiation can occur by the recruitment of additional postsynaptic AMPA receptors (AMPARs), sourced either from an intracellular reserve pool by exocytosis or from nearby extra-synaptic receptors pre-existing on the neuronal surface. However, the exact mechanism through which synapses can rapidly recruit new AMPARs during early LTP remains unknown. In particular, direct evidence for a pivotal role of AMPAR surface diffusion as a trafficking mechanism in synaptic plasticity is still lacking. Here, using AMPAR immobilization approaches, we show that interfering with AMPAR surface diffusion markedly impairs synaptic potentiation of Schaffer collaterals and commissural inputs to the CA1 area of the mouse hippocampus in cultured slices, acute slices and in vivo. Our data also identify distinct contributions of various AMPAR trafficking routes to the temporal profile of synaptic potentiation. In addition, AMPAR immobilization in vivo in the dorsal hippocampus inhibited fear conditioning, indicating that AMPAR diffusion is important for the early phase of contextual learning. Therefore, our results provide a direct demonstration that the recruitment of new receptors to synapses by surface diffusion is a critical mechanism for the expression of LTP and hippocampal learning. Since AMPAR surface diffusion is dictated by weak Brownian forces that are readily perturbed by protein-protein interactions, we anticipate that this fundamental trafficking mechanism will be a key target for modulating synaptic potentiation and learning.

Zinc and copper ions can modulate the activity of glutamate receptors. However, labile zinc and copper ions likely represent only the tip of the iceberg and other neuronal functions are suspected for these metals in their bound state. We performed synchrotron X-ray fluorescence imaging with 30 nm resolution to image total biometals in dendrites and spines from hippocampal neurons. We found that zinc is distributed all along the dendrites while copper is mainly pinpointed within the spines. In spines, zinc content is higher within the spine head while copper is higher within the spine neck. Such specific distributions suggested metal interactions with cytoskeleton proteins. Zinc supplementation induced the increase of β-tubulin content in dendrites. Copper supplementation impaired the β-tubulin and F-actin networks. Copper chelation resulted in the decrease of F-actin content in dendrites, drastically reducing the number of F-actin protrusions. These results indicate that zinc is involved in microtubule stability whereas copper is essential for actin-dependent stability of dendritic spines, although copper excess can impair the dendritic cytoskeleton.

Depression and dementia are commonly concurrent and are both associated with increased mortality among older people. However, little is known about whether home-dwelling patients newly diagnosed with mild dementia coexisting with depressive symptoms have excess mortality. We conducted a post hoc analysis based on data from the Danish Alzheimer's Intervention Study of 330 individuals who were diagnosed with mild dementia within the past 12 months. Thirty-four patients were identified with major depressive symptoms (MD-S) at baseline. During the 3-year follow-up period, 56 patients died, and, among them, 12 were with MD-S at baseline. Multivariable analysis adjusting for the potential confounders (age, sex, smoking status, alcohol consumption, education, BMI, household status, MMSE, CCI, QoL-AD, NPIQ, ADSC-ADL, medication, and RCT allocation) showed that patients with MD-S had a 2.5-fold higher mortality as compared to the patients without or with only few depressive symptoms. Our result revealed that depression is possibly associated with increased mortality in patients with mild dementia. Given that depression is treatable, screening for depression and treatment of depression can be important already in the earliest stage of dementia to reduce mortality.

Objective: The purpose of this study was to examine if parental separation during childhood is associated with obesity in adulthood., Methods: A co-twin case-control study of 146 adult same-sexed twin pairs with discordant body mass index (BMI) (i.e. one of the twins should have a BMI of 20-25 kg/m 2 , and the co-twin's BMI ≥ 30 kg/m 2 ) was selected from Danish Twin Registry (DTR). In total of 236 eligible twin individuals participated in the study. Childhood parental separation (defined as separation from one of the biological parents, regardless of the reason for separation) for at least one year prior to age 17 was self-reported. The statistical analysis includes logistic and linear regression models using STATA 13.0., Results: There were no differences in the odds of developing obesity in adulthood between the twin who stayed with a father and the co-twin who was separated from him for at least 1 year prior to age 17 [OR = 1.22, 95%CI (0.46-3.34), p  = 0.65]. Separation from a mother also showed no differences in the odds for developing obesity [OR = 0.90, 95%CI (0.32-2.46), p  = 0.82]., Conclusions: Because of the limited number of discordant twin pairs for childhood parental separation, we cannot provide evidence to suggest that separation from parents in childhood was associated with developing obesity in adulthood. Further studies of pooling discordant twins from several countries should be considered.

Background: As a consequence of a rapid growth of an ageing population, more people with dementia are expected on the roads. Little is known about whether these people are at increased risk of road traffic-related accidents., Objective: Our study aims to investigate the risk of road traffic-related accidents for people aged 65 years or older with a diagnosis of dementia in Denmark., Methods: We will conduct a nationwide population-based cohort study consisting of Danish people aged 65 or older living in Denmark as of January 1, 2008. The cohort is followed for 7 years (2008-2014). Individual's personal data are available in Danish registers and can be linked using a unique personal identification number. A person is identified with dementia if the person meets at least one of the following criteria: (1) a diagnosis of the disease in the Danish National Patient Register or in the Danish Psychiatric Central Research Register, and/or (2) at least one dementia diagnosis-related drug prescription registration in the Danish National Prescription Registry. Police-, hospital-, and emergency room-reported road traffic-related accidents occurred within the study follow-up are defined as the study outcome. Cox proportional hazard regression models are used for the main analysis., Results: Our study protocol has 3 phases including data collection, data analysis, and reporting. The first phase of register-based data collection of 853,228 individual's personal information was completed in August, 2016. The next phase is data analysis, which is expected to be finished before December 2016, and thereafter writing publications based on the findings. The study started in January 2016 and will end in December 2018., Discussion: This study covers the entire elderly population of Denmark, and thereby will avoid selection bias due to nonparticipation and loss to follow-up. Furthermore, this ensures that the study results are reliable and generalizable. However, underreporting of traffic-related accidents may occur, which will limit estimation of absolute risks., Competing Interests: Conflicts of Interest: None declared.

Trafficking and biophysical properties of AMPA receptors (AMPARs) in the brain depend on interactions with associated proteins. We identify Shisa6, a single transmembrane protein, as a stable and directly interacting bona fide AMPAR auxiliary subunit. Shisa6 is enriched at hippocampal postsynaptic membranes and co-localizes with AMPARs. The Shisa6 C-terminus harbours a PDZ domain ligand that binds to PSD-95, constraining mobility of AMPARs in the plasma membrane and confining them to postsynaptic densities. Shisa6 expressed in HEK293 cells alters GluA1- and GluA2-mediated currents by prolonging decay times and decreasing the extent of AMPAR desensitization, while slowing the rate of recovery from desensitization. Using gene deletion, we show that Shisa6 increases rise and decay times of hippocampal CA1 miniature excitatory postsynaptic currents (mEPSCs). Shisa6-containing AMPARs show prominent sustained currents, indicating protection from full desensitization. Accordingly, Shisa6 prevents synaptically trapped AMPARs from depression at high-frequency synaptic transmission.

Landscape-scale intensification of individual crops and pesticide use that is associated with this intensification is an emerging, environmental problem that is expected to have unequal effects on pests with different lifecycles, host ranges, and dispersal abilities. We investigate if intensification of a single crop in an agroecosystem has a direct effect on insecticide resistance in a specialist insect herbivore. Using a major potato pest, Leptinotarsa decemlineata, we measured imidacloprid (neonicotinoid) resistance in populations across a spatiotemporal crop production gradient where potato production has increased in Michigan and Wisconsin, USA. We found that concurrent estimates of area and temporal frequency of potato production better described patterns of imidacloprid resistance among L. decemlineata populations than general measures of agricultural production (% cropland, landscape diversity). This study defines the effects individual crop rotation patterns can have on specialist herbivore insecticide resistance in an agroecosystem context, and how impacts of intensive production can be estimated with general estimates of insecticide use. Our results provide empirical evidence that variation in the intensity of neonicotinoid-treated potato in an agricultural landscape can have unequal impacts on L. decemlineata insecticide insensitivity, a process that can lead to resistance and locally intensive insecticide use. Our study provides a novel approach applicable in other agricultural systems to estimate impacts of crop rotation, increased pesticide dependence, insecticide resistance, and external costs of pest management practices on ecosystem health.

Excitatory glutamatergic synapses at dendritic spines exchange and modulate their receptor content via lateral membrane diffusion. Several studies have shown that the thin spine neck impedes the access of membrane and solute molecules to the spine head. However, it is unclear whether the spine neck geometry alone restricts access to dendritic spines or if a physical barrier to the diffusion of molecules exists. Here, we investigated whether a complex of septin cytoskeletal GTPases localized at the base of the spine neck regulates diffusion across the spine neck. We found that, during development, a marker of the septin complex, Septin7 (Sept7), becomes localized to the spine neck where it forms a stable structure underneath the plasma membrane. We show that diffusion of receptors and bulk membrane, but not cytoplasmic proteins, is slower in spines bearing Sept7 at their neck. Finally, when Sept7 expression was suppressed by RNA interference, membrane molecules explored larger membrane areas. Our findings indicate that Sept7 regulates membrane protein access to spines.

Potential decreases in crop yield from reductions in bee-mediated pollination services threaten food production demands of a growing population. Many fruit and vegetable growers supplement their fields with bee colonies during crop bloom. The extent to which crop production requires supplementary pollination services beyond those provided by wild bees is not well documented. Pumpkin, Cucurbita pepo L., requires bee-mediated pollination for fruit development. Previous research identified the common eastern bumble bee, Bombus impatiens (Cresson), as the most efficient pumpkin pollinator. Two concomitant studies were conducted to examine pollination deficits in New York pumpkin fields from 2011 to 2013. In the first study, fruit weight, seed set, and B. impatiens visits to pumpkin flowers were compared across fields supplemented with B. impatiens colonies at a recommended stocking density of five colonies per hectare, a high density of 15 colonies per hectare, or not supplemented with bees. In the second study, fruit weight and seed set of pumpkins that received supplemental pollen through hand-pollination were compared with those that were open-pollinated by wild bees. Results indicated that supplementing pumpkin fields with B. impatiens colonies, regardless of stocking density, did not increase fruit weight, seed set, or B. impatiens visits to pumpkin flowers. Fruit weight and seed set did not differ between hand- and open-pollinated treatments. In general, we conclude that pumpkin production in central New York is not limited by inadequate pollination services provided by wild bees and that on average, supplementation with B. impatiens colonies did not improve pumpkin yield.

The polarized distribution of membrane proteins to axonal or somatodendritic neuronal compartments is fundamental to nearly every aspect of neuronal function. The polarity of dendritic proteins depends on selective microtubule-based transport; the vesicles that carry these proteins are transported into dendrites but do not enter the axon. We used live-cell imaging of fluorescently tagged dendritic and axonal proteins combined with immunostaining for initial segment and cytoskeletal markers to evaluate different models of dendrite-selective transport in cultured rat hippocampal neurons. In mature neurons, dendritic vesicles that entered the base of the axon stopped at the proximal edge of the axon initial segment, defined by immunostaining for ankyrinG, rather than moving into the initial segment itself. In contrast, axonal vesicles passed through the initial segment without impediment. During development, dendrite-selective transport was detected shortly after axons formed, several days before initial segment assembly, before the appearance of a dense actin meshwork in the initial segment, and before dendrites acquire microtubules of mixed polarity orientation. Indeed, some elements of selective transport were detected even before axon specification. These findings are inconsistent with models for selective transport that depend on the presence of an F-actin-based cytoplasmic filter in the initial segment or that posit that transport into dendrites is mediated by dyneins translocating along minus-end out microtubules. Instead our results suggest that selective transport involves the coordinated regulation of the different motor proteins that mediate dendritic vesicle transport and that the selectivity of motor-microtubule interactions is one facet of this process.