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Abstract Functional analysis of non-coding variants associated with congenital disorders remains challenging due to the lack of efficient in vivo models. Here we introduce dual-enSERT, a robust Cas9-based two-color fluorescent reporter system which enables rapid, quantitative comparison of enhancer allele activities in live mice in less than two weeks. We use this technology to examine and measure the gain- and loss-of-function effects of enhancer variants previously linked to limb polydactyly, autism spectrum disorder, and craniofacial malformation. By combining dual-enSERT with single-cell transcriptomics, we characterise gene expression in cells where the enhancer is normally and ectopically active, revealing candidate pathways that may lead to enhancer misregulation. Finally, we demonstrate the widespread utility of dual-enSERT by testing the effects of fifteen previously uncharacterised rare and common non-coding variants linked to neurodevelopmental disorders. In doing so we identify variants that reproducibly alter the in vivo activity of OTX2 and MIR9-2 brain enhancers, implicating them in autism. Dual-enSERT thus allows researchers to go from identifying candidate enhancer variants to analysis of comparative enhancer activity in live embryos in under two weeks.

Abstract Objective Black American (BA) multiple sclerosis (MS) patients experience greater disability compared to White American (WA) patients. Here, we investigated the role of paramagnetic rim lesions (PRLs), a subset of chronic active lesions, on race‐related disability in MS. Methods We conducted a retrospective observational study comparing BA and WA MS patients. PRLs were identified through Quantitative Susceptibility Mapping (QSM) MRI. A causal mediation analysis explored the impact of PRLs on the relationship between race and disability, as measured by the Expanded Disability Status Scale (EDSS). Results The prevalence of PRLs in BA patients with MS was higher at 55% compared to WA patients at 39% (p = 0.022). A higher percentage of PRLs among all white matter lesions was observed with BA (8.01%) patients compared to WA (3.4%) patients (p = 0.003). In a regression analysis, controlling for significant patient‐level covariates and income‐level demographics, the percentage of PRLs was, on average, 4.61 points higher for BA patients than for WA patients (p = 0.003). In a separate regression analysis, accounting for covariates, BA patients exhibited significantly higher EDSS scores (p

Abstract Background Common mental health problems, such as stress, anxiety and depression, are highly prevalent among workers and often lead to long-term absenteeism and work disability. Effective elements found in previous researched interventions were to explicitly focus on return to work (RTW) and not solely on symptom reduction, to take into account the employees’ cognition towards RTW and to include the workplace environment. Based on these elements, a stepped-care approach was developed. The aim of this paper is to present the study design of a randomized controlled trial (RESTART), evaluating the effectiveness of the stepped-care approach on lasting RTW and the implementation process. Methods RESTART is a randomized controlled trial with a 2 × 2 factorial design and a follow-up of one year. Employees eligible for this study are those who reported sick within 2 to 8 weeks with psychological distress based on a distress screener. Participants will be randomized to a group receiving a tailored e-Health app or usual care, as well as randomized to a group receiving a Participatory Approach (PA; conversational method) in the workplace or usual care. The PA will however only be provided in case of persistent sickness absence at 8 weeks. Measurements take place at baseline, after the e-Health intervention period (3 months), and after the PA intervention period (6 months) and 12 months. Primary outcome is lasting RTW, defined as full RTW in previous or equal work for at least four consecutive weeks. Secondary outcomes are (the severity of) stress-related symptoms, total number of sickness absence days, self-efficacy for RTW and self-reported health. A process evaluation including a realist evaluation will also be conducted. Discussion Early intervention that focuses on RTW, the cognition towards RTW despite symptoms and involves the workplace environment, plays a crucial role in managing sickness absence among employees with psychological distress. If effective, the stepped-care approach is relevant for employees, employers and society as a whole. Trial Registration ISRCTN: 90663076. Registered on 5 October 2023.

White striping (WS) is an anomaly characterized by the appearance of white striations in the Pectoralis major muscles. This study aimed to evaluate the effect of animal fat replacement (chicken skin) with hydrolyzed collagen (HC, 0, 50 and 75%) on the characteristics of burgers with normal chicken breast fillets (FN) and WS. The samples were subjected to physical-chemical characterization, oxidative stability (TBARS), texture, and sensory profile. The utilization of WS meat increased the pH values, lipid content, and oxidation throughout storage, but it did not impair the sensory acceptance and sensory profile of the products. Adding HC reduced the pH in the formulations, changed the chemical composition (lower moisture and lipid content and higher protein and collagen content), and increased the texture parameters (hardness and chewiness). The formulations with 50% fat replacement showed similar sensory acceptance and purchase intention to the control products and were characterized by the typical texture and mild aroma and flavor. The results suggest that WS chicken meat and HC can be used to produce hamburgers with 50% replacement of animal fat without compromising the quality of the products.

The mechanisms of initiation of spreading depolarization (SD) are understudied due to a paucity of disease models with spontaneously occurring events. We here present a novel mouse model of familial hemiplegic migraine type 2 (FHM2), expressing the missense T345A-mutated α2 subunit of the Na+/K+ adenosine triphosphatase pump (Atp1a2T345A). Homozygous Atp1a2T345A mice showed regular spontaneous SDs that exhibit a diurnal rhythm and typically originate from the hippocampus. Heterozygous Atp1a2T345A mice rarely exhibited spontaneous SDs and, for electrically induced SDs, only showed an increased propagation speed, whereas homozygotes showed both increased propagation and decreased threshold. Remarkably, despite hippocampal hyperexcitability, spontaneous SDs in Atp1a2T345A mice were only rarely associated with epileptic behavior, and seizure expression during kindling was decreased. Spontaneous SDs could be prevented by modulation of persistent sodium currents. Hippocampal SDs occurred in the presence of an NMDA-receptor antagonist, but these events did not reach the cortex, suggesting that initiation and propagation of SD depend on different mechanisms in this model.

ObjectivesWhite collar workers spend an increasing amount of time in occupational sedentary behavior (OSB) and are thereby at risk for adverse health outcomes. Nevertheless, the association between OSB and the need for recovery (NFR), an important indicator of wellbeing, is unknown and therefore examined.MethodsBaseline data from a cluster randomized controlled trial was used. A subgroup of 89 white collar workers wore a triaxial accelerometer for 7 days. NFR was measured using the Questionnaire on the Experience and Evaluation of Work. Compositional data analysis was applied to determine the composition of different OSB bouts (short, medium and long) and occupational physical activity (OPA) (light, moderate and vigorous and standing). Linear regression analyses were performed to explore the associations between occupational compositions and NFR.ResultsRelatively more time spent in long OSB bouts was associated with a lower NFR (β: −11.30, 95% CI: −20.2 to −2.4). Short and medium OSB bouts and OPA were not associated with NFR.ConclusionAssociations between OSB bouts, OPA and NFR hinted at contrasting trends, suggesting the need to consider different bout lengths of OSB in future studies.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease, involving the selective degeneration of cortical upper synapses in the primary motor cortex (M1). Excitotoxicity in ALS occurs due to an imbalance between excitation and inhibition, closely linked to the loss/gain of astrocytic function. Using the ALS SOD1G93A mice, we investigated the astrocytic contribution for the electrophysiological alterations observed in the M1 of SOD1G93A mice, throughout disease progression. Results showed that astrocytes are involved in synaptic dysfunction observed in presymptomatic SOD1G93A mice, since astrocytic glutamate transport currents are diminished and pharmacological inhibition of astrocytes only impaired long-term potentiation and basal transmission in wild-type mice. Proteomic analysis revealed major differences in neuronal transmission, metabolism, and immune system in upper synapses, confirming early communication deficits between neurons and astroglia. These results provide valuable insights into the early impact of upper synapses in ALS and the lack of supportive functions of cortical astrocytes, highlighting the possibility of manipulating astrocytes to improve synaptic function.

Mechanisms underlying the migraine aura are incompletely understood, which to large extent is related to a lack of models in which cortical spreading depolarization (CSD), the correlate of the aura, occurs spontaneously. Here, we investigated electrophysiological and behavioural CSD features in freely behaving mice expressing mutant CaV2.1 Ca2+ channels, either with the milder R192Q or the severer S218L missense mutation in the α1 subunit, known to cause familial hemiplegic migraine type 1 (FHM1) in patients. Very rarely, spontaneous CSDs were observed in mutant but never in wildtype mice. In homozygous Cacna1aR192Q mice exclusively single-wave CSDs were observed whereas heterozygous Cacna1aS218L mice displayed multiple-wave events, seemingly in line with the more severe clinical phenotype associated with the S218L mutation. Spontaneous CSDs were associated with body stretching, one-directional slow head turning, and rotating movement of the body. Spontaneous CSD events were compared with those induced in a controlled manner using minimally invasive optogenetics. Also in the optogenetic experiments single-wave CSDs were observed in Cacna1aR192Q and Cacna1aS218L mice (whereas the latter also showed multiple-wave events) with movements similar to those observed with spontaneous events. Compared to wildtype mice, FHM1 mutant mice exhibited a reduced threshold and an increased propagation speed for optogenetically induced CSD with a more profound CSD-associated dysfunction, as indicated by a prolonged suppression of transcallosal evoked potentials and a reduction of unilateral forepaw grip performance. When induced during sleep, the optogenetic CSD threshold was particularly lowered, which may explain why spontaneous CSD events predominantly occurred during sleep. In conclusion, our data show that key neurophysiological and behavioural features of optogenetically induced CSDs mimic those of rare spontaneous events in FHM1 R192Q and S218L mutant mice with differences in severity in line with FHM1 clinical phenotypes seen with these mutations.

Abstract Background Cortical spreading depolarization (CSD), the neurophysiological correlate of the migraine aura, can activate trigeminal pain pathways, but the neurobiological mechanisms and behavioural consequences remain unclear. Here we investigated effects of optogenetically-induced CSDs on headache-related behaviour and neuroinflammatory responses in transgenic mice carrying a familial hemiplegic migraine type 1 (FHM1) mutation. Methods CSD events (3 in total) were evoked in a minimally invasive manner by optogenetic stimulation through the intact skull in freely behaving wildtype (WT) and FHM1 mutant mice. Related behaviours were analysed using mouse grimace scale (MGS) scoring, head grooming, and nest building behaviour. Neuroinflammatory changes were investigated by assessing HMGB1 release with immunohistochemistry and by pre-treating mice with a selective Pannexin-1 channel inhibitor. Results In both WT and FHM1 mutant mice, CSDs induced headache-related behaviour, as evidenced by increased MGS scores and the occurrence of oculotemporal strokes, at 30 min. Mice of both genotypes also showed decreased nest building behaviour after CSD. Whereas in WT mice MGS scores had normalized at 24 h after CSD, in FHM1 mutant mice scores were normalized only at 48 h. Of note, oculotemporal stroke behaviour already normalized 5 h after CSD, whereas nest building behaviour remained impaired at 72 h; no genotype differences were observed for either readout. Nuclear HMGB1 release in the cortex of FHM1 mutant mice, at 30 min after CSD, was increased bilaterally in both WT and FHM1 mutant mice, albeit that contralateral release was more pronounced in the mutant mice. Only in FHM1 mutant mice, contralateral release remained higher at 24 h after CSD, but at 48 h had returned to abnormal, elevated, baseline values, when compared to WT mice. Blocking Panx1 channels by TAT-Panx308 inhibited CSD-induced headache related behaviour and HMGB1 release. Conclusions CSDs, induced in a minimally invasive manner by optogenetics, investigated in freely behaving mice, cause various migraine relevant behavioural and neuroinflammatory phenotypes that are more pronounced and longer-lasting in FHM1 mutant compared to WT mice. Prevention of CSD-related neuroinflammatory changes may have therapeutic potential in the treatment of migraine.

Astrocytes, the most abundant glial cells in the central nervous system (CNS), sense synaptic activity and respond through the release of gliotransmitters, a process mediated by intracellular Ca2+ level changes and SNARE-dependent mechanisms. Ionotropic N-methyl-D-aspartate (NMDA) receptors, which are activated by glutamate along with D-serine or glycine, play a crucial role in learning, memory, and synaptic plasticity. However, the precise impact of astrocyte-released D-serine on neuronal modulation remains insufficiently characterized. To address this, we have used the dominant negative SNARE (dnSNARE) mouse model, which selectively inhibits SNARE-dependent exocytosis from astrocytes. We recorded field excitatory postsynaptic potentials (fEPSPs) in CA3-CA1 synapses within hippocampal slices obtained from dnSNARE mice and wild-type (Wt) littermates. Our results demonstrate that hippocampal θ-burst long-term potentiation (LTP), a critical form of synaptic plasticity, is impaired in hippocampal slices from dnSNARE mice. Notably, this LTP impairment was rescued upon incubation with D-serine. To further investigate the involvement of astrocytes in D-serine-mediated mechanisms of LTP maintenance, we perfused hippocampal slices with L-serine – a substrate used by both neurons and astrocytes for D-serine production. The enhancement in LTP observed in dnSNARE mice was exclusively associated with D-serine presence, with no effects evident in the presence of L-serine. Additionally, both D- and L-serine reduced basal synaptic strength in the hippocampal slices of both Wt and dnSNARE mice. These results provide compelling evidence that distinct processes underlie the modulation of basal synaptic transmission and LTP through D-serine. Our findings underscore the pivotal contribution of astrocytes in D-serine-mediated processes that govern LTP establishment and basal transmission. This study not only provides essential insights into the intricate interplay between neurons and astrocytes but also emphasizes their collective role in shaping hippocampal synaptic function.

IntroductionBurn-out leads to reduced worker well-being, long-term absenteeism, and high costs for employers and society. Determinants at different levels may affect burn-out in an interrelated and dynamic manner. The aim of the present study was to apply a broader systems perspective by exploring and visualizing the complex system of determinants at different levels (living conditions, working conditions, and societal developments) underlying the prevalence of burn-out in the Netherlands.MethodsDuring three group model building (GMB) sessions with in total eight experts on workers’ mental health, a causal loop diagram (CLD) was developed and relevant feedback loops were identified. For the selection of determinants to be included in the CLD a recently published overview of determinants on burn-out at different levels was used. Experts could also add factors that were not listed in the overview.ResultsThe final CLD consists of 20 factors and depicts a central position of working conditions. Societal developments (e.g., access to mental health care, size of the working population, rougher social climate, etc.) were mostly located at the outside of the CLD and barely integrated in feedback loops. Several reinforcing feedback loops resulting in an increase of the prevalence of burn-out were identified in which the factors (very) high workload, imbalance between work and private life, and insufficient recovery time play an important role. Also, several balancing loops were found that visualize the crucial role of functional support from supervisors to prevent burn-out among workers.DiscussionApplying a broader systems perspective, including determinants at different levels, offers new insights into dynamic feedback loops that contribute to the prevalence of burn-out. Supervisors, amongst others, have a considerable impact on the system underlying the high prevalence of burn-out and may therefore contribute to its prevention. Even though societal developments were less integrated in feedback loops, they might be considered drivers of existing feedback loops. The results from this study confirm that determinants at various levels underly the prevalence of burn-out. To be able to address the diversity of determinants underlying a high prevalence of burn-out, a complex system approach can be helpful.

PurposeIn March 2020, the WHO declared COVID-19 a pandemic. Previous virus outbreaks, such as the SARS outbreak in 2003, appeared to have a great impact on the mental health of healthcare workers. The aim of this study is to examine to what extent mental health of healthcare workers differed from non-healthcare workers during the first year of the COVID-19 pandemic.MethodsWe used data from a large-scale longitudinal online survey conducted by the Corona Behavioral Unit in the Netherlands. Eleven measurement rounds were analyzed, from April 2020 to March 2021 (N = 16,615; number of observations = 64,206). Mental health, as measured by the 5-item Mental Health Inventory, was compared between healthcare workers and non-healthcare workers over time, by performing linear GEE-analyses.ResultsMental health scores were higher among healthcare workers compared to non-healthcare workers during the first year of the pandemic (1.29 on a 0–100 scale, 95%-CI = 0.75–1.84). During peak periods of the pandemic, with over 100 hospital admissions or over 25 ICU admissions per day and subsequently more restrictive measures, mental health scores were observed to be lower in both healthcare workers and non-healthcare workers.ConclusionDuring the first year of the COVID-19 pandemic, we observed no relevant difference in mental health between healthcare workers and non-healthcare workers in the Netherlands. To be better prepared for another pandemic, future research should investigate which factors hinder and which factors support healthcare workers to maintain a good mental health.

BackgroundDuring the COVID-19 pandemic, many healthcare workers faced extreme working conditions and were at higher risk of infection with the coronavirus. These circumstances may have led to mental health problems, such as anxiety, among healthcare workers. Most studies that examined anxiety among healthcare workers during the COVID-19 pandemic were cross-sectional and focused on the first months of the pandemic only. Therefore, this study aimed to investigate the longitudinal association between working in healthcare and anxiety during a long-term period (i.e., 18 months) of the COVID-19 pandemic.MethodsData were used from online questionnaires of the Lifelines COVID-19 prospective cohort with 22 included time-points (March 2020–November 2021). In total, 2,750 healthcare workers and 9,335 non-healthcare workers were included. Anxiety was assessed with questions from the Mini-International Neuropsychiatric Interview, and an anxiety sum score (0–7) was calculated. Negative binomial generalized estimating equations (GEE), adjusted for demographic, work and health covariates, were used to examine the association between working in healthcare and anxiety.ResultsAnxiety sum scores over time during the COVID-19 pandemic were similar for healthcare workers and non-healthcare workers. No differences between the anxiety sum scores of healthcare workers and non-healthcare workers were found [incidence rate ratio (IRR) = 0.97, 95% CI = 0.91–1.04].ConclusionThis study did not find differences between healthcare workers and non-healthcare in perceived anxiety during the COVID-19 pandemic.

Introducción: La disciplina Ergonomía/Factores Humanos es demandada por los empresarios que se percatan de su utilidad, no solo para mejorar la salud de los empleados; sino porque constituye una herramienta útil para alcanzar mayores niveles productivos, de calidad y competitividad muy necesarios en la actualidad. Objetivo: Evaluar desde el punto de vista ergonómico uno de los puestos donde se desarrolla la producción de mangueras hidráulicas y de freno hidráulico solicitadas por las empresas azucareras, estableciendo los aspectos que influyen directamente en los resultados productivos. Materiales y Métodos: Para diagnosticar la situación se hizo uso de un procedimiento validado en otras situaciones para el análisis ergonómico que reúne un conjunto de fases y etapas asociadas a técnicas que posibilitan reportar los inadecuados niveles de diseño en el puesto de trabajo. Resultados y Discusión: Los principales resultados mostraron que factores relacionados con variables ergonómicas inciden en el diseño y organización del puesto analizado y determinan en los niveles productivos obtenidos. Además, fue posible identificar las afectaciones que produce el puesto sobre la operaria. Conclusiones: La implementación del programa ergonómico diseñado personalizado a las necesidades existentes puede contribuir a mejorar los niveles productivos, la calidad de vida laboral y salud de la trabajadora.

Typical absence seizures (ASs) are brief periods of lack of consciousness, associated with 2.5–4 Hz spike-wave discharges (SWDs) in the EEG, which are highly prevalent in children and teenagers. The majority of probands in these young epileptic cohorts show neuropsychological comorbidities, including cognitive, memory and mood impairments, even after the seizures are pharmacologically controlled. Similar cognition and memory deficits have been reported in different, but not all, genetic animal models of ASs. However, since these impairments are subtle and highly task-specific their presence may be confounded by an anxiety-like phenotype and no study has tested anxiety and memory in the same animals. Moreover, the majority of studies used non-epileptic inbred animals as the only control strain and this may have contributed to a misinterpretation of these behavioural results. To overcome these issues, here we used a battery of behavioural tests to compare anxiety and memory in the same animals from the well-established inbred model of Genetic Absence Epilepsy Rats from Strasbourg (GAERS), their inbred strain of Non-Epileptic Control (NEC) strain (that lack ASs) and normal outbred Wistar rats. We found that GAERS do not exhibit increased anxiety-like behavior and neophobia compared to both NEC and Wistar rats. In contrast, GAERS show decreased spontaneous alternation, spatial working memory and cross-modal object recognition compared to both NEC and Wistar rats. Furthermore, GAERS preferentially used egocentric strategies to perform spatial memory tasks. In summary, these results provide solid evidence of memory deficits in GAERS rats that do not depend on an anxiety or neophobic phenotype. Moreover, the presence of differences between NEC and Wistar rats stresses the need of using both outbred and inbred control rats in behavioural studies involving genetic models of ASs.

Non-epileptic seizures are identified as a common epileptogenic trigger. Early metaplasticity following seizures may contribute to epileptogenesis by abnormally altering synaptic strength and homeostatic plasticity. We now studied how in vitro epileptiform activity (EA) triggers early changes in CA1 long-term potentiation (LTP) induced by theta-burst stimulation (TBS) in rat hippocampal slices and the involvement of lipid rafts in these early metaplasticity events. Two forms of EA were induced: (1) interictal-like EA evoked by Mg2+ withdrawal and K+ elevation to 6 mM in the superfusion medium or (2) ictal-like EA induced by bicuculline (10 μM). Both EA patterns induced and LTP-like effect on CA1 synaptic transmission prior to LTP induction. LTP induced 30 min post EA was impaired, an effect more pronounced after ictal-like EA. LTP recovered to control levels 60 min post interictal-like EA but was still impaired 60 min after ictal-like EA. The synaptic molecular events underlying this altered LTP were investigated 30 min post EA in synaptosomes isolated from these slices. EA enhanced AMPA GluA1 Ser831 phosphorylation but decreased Ser845 phosphorylation and the GluA1/GluA2 ratio. Flotillin-1 and caveolin-1 were markedly decreased concomitantly with a marked increase in gephyrin levels and a less prominent increase in PSD-95. Altogether, EA differentially influences hippocampal CA1 LTP thorough regulation of GluA1/GluA2 levels and AMPA GluA1 phosphorylation suggesting that altered LTP post-seizures is a relevant target for antiepileptogenic therapies. In addition, this metaplasticity is also associated with marked alterations in classic and synaptic lipid raft markers, suggesting these may also constitute promising targets in epileptogenesis prevention.

OBJECTIVE: Working from home during the COVID-19 pandemic has affected many workers’ daily life and possibly their physical activity behavior. We studied the longitudinal association of working from home during the pandemic with physical activity and sedentary behavior. METHODS: Longitudinal data from 17 questionnaire rounds of the Lifelines COVID-19 cohort (March 2020–February 2021) were used. In total, 33 325 workers were included. In every round, participants reported their current work situation: location, home, or hybrid (working on location and from home). Physical activity levels and sedentary behavior before and during the pandemic were asked. Logistic generalized estimating equations adjusted for demographic/work/health covariates were used to study the association of work situation with physical activity and sedentary behavior. RESULTS: Home workers were less likely to meet the recommended ≥150 minutes/week of moderate-to-vigorous-intensity activity during the pandemic than location workers [odds ratio (OR) 0.93, 95% confidence interval (CI) 0.90–0.96] and more likely to be less physically active than before the pandemic (OR 1.09, 95% CI 1.04–1.14). Furthermore, compared to location workers, home and hybrid workers were more likely to be more sedentary (sitting ≥8 hours/day) on workdays during than before the pandemic (OR 1.51, 95% CI 1.39–1.64/1.36–1.68, respectively). CONCLUSIONS: Compared to location workers, home workers (and to a lesser extent hybrid workers) were more often physically inactive and sedentary during than before the COVID-19 pandemic. As a substantial part of the working population may continue to work (partly) from home after the pandemic, workers should be supported to increase activity and reduce sitting while working from home.

Abstract Background To explore whether differences between men and women in the sensitivity to (strength of the association) and/or in the exposure to determinants (prevalence) contribute to the difference in physical functioning, with women reporting more limitations. Methods Data of the Doetinchem Cohort Study was used (n = 5856, initial ages 26–70 years), with follow-up measurements every 5 years (up to 20). Physical functioning (subscale SF-36, range:0–100), sex (men or women) and a number of socio-demographic, lifestyle- and health-related determinants were assessed. Mixed-model multivariable analysis was used to investigate differences between men and women in sensitivity (interaction term with sex) and in exposure (change of the sex difference when adjusting) to determinants of physical functioning. Results The physical functioning score among women was 6.55 (95%CI:5.48,7.61) points lower than among men. In general, men and women had similar determinants, but pain was more strongly associated with physical functioning (higher sensitivity), and also more prevalent among women (higher exposure). The higher exposure to low educational level and not having a paid job also contributed to the lower physical functioning score among women. In contrast, current smoking, mental health problems and a low educational level were more strongly associated with a lower physical functioning score among men and lower physical activity and higher BMI were more prevalent among men. Conclusions Although important for physical functioning among both men and women, our findings provide no indications for reducing the difference in physical functioning by promoting a healthy lifestyle but stress the importance of differences in pain, work and education.

Abstract Background An integrated workplace health promotion program (WHPP) which targets multiple lifestyle factors at different levels (individual and organizational) is potentially more effective than a single component WHPP. The aim of this study is to describe the protocol of a study to tailor a European good practice of such an integral approach to the Dutch context and to evaluate its effectiveness and implementation. Methods This study consists of two components. First, the five steps of the Map of Adaptation Process (MAP) will be followed to tailor the Lombardy WHP to the Dutch context. Both the employers and employees will be actively involved in this process. Second, the effectiveness of the integrated Dutch WHPP will be evaluated in a clustered randomized controlled trial (C-RCT) with measurements at baseline, 6 months and 12 months. Clusters will be composed based on working locations or units - dependent on the organization’s structure and randomization within each organization takes place after baseline measurements. Primary outcome will be a combined lifestyle score. Secondary outcomes will be the separate lifestyle behaviors targeted, stress, work-life balance, need for recovery, general health, and well-being. Simultaneously, a process evaluation will be conducted. The study population will consist of employees from multiple organizations in different industry sectors. Organizations in the intervention condition will receive the integrated Dutch WHPP during 12 months, consisting of an implementation plan and a catalogue with activities for multiple lifestyle themes on various domains: 1) screening and support; 2) information and education; 3) adjustments in the social, digital or physical environment; and 4) policy. Discussion The MAP approach provides an appropriate framework to systematically adapt an existing WHPP to the Dutch context, involving both employers and employees and retaining the core elements, i.e. the catalogue with evidence-based activities on multiple lifestyle themes and domains enabling an integrated approach. The following process and effect evaluation will contribute to further insight in the actual implementation and effectiveness of the integrated WHP approach. Trial registration NTR (trialregister.nl ), NL9526. Registered on 3 June 2021.

BackgroundWorkplace health promotion programs (WHPPs) can benefit the lifestyle and health of employees. However, not all WHPPs have been successful in their implementation, and thus their effectiveness. This study aimed to identify the barriers and facilitators to implementing an integrated WHPP, which targets multiple lifestyle factors at different levels (individual and organizational), from an employer's perspective.MethodsData were collected by two online focus groups among 18 representatives of eight different organizations. Data from the focus group discussions were transcribed verbatim and analyzed using thematic analysis. Data were coded both inductively and deductively, using the Consolidated Framework for Implementation Research (CFIR) consisting of the following five domains: (1) intervention characteristics, (2) outer setting, (3) inner setting, (4) characteristics of individuals, and (5) process. Ratings were performed to indicate the positive or negative influence and strength of a construct regarding the implementation of WHPPs.ResultsBarriers and facilitators in all domains of the CFIR were found. Regarding characteristics of the WHPP, complexity and costs hindered implementation, while high adaptability facilitated it. An organization that met the needs of employees (the outer setting) facilitated implementation. Available resources, access to knowledge, leadership involvement, and continuity of communication were facilitators within the inner setting. Barriers were different approaches to implementation within one organization and the perceived interference with employees' lives. For the implementation process, the involvement of key stakeholders, including employees, was identified as an important facilitator.ConclusionVarious barriers and facilitators in different domains play a role in the implementation of integrated WHPPs, according to employers. Strategies that tackle the identified barriers and incorporate the facilitators will likely contribute to the successful implementation of integrated WHPPs.

Despite a substantial evidence base for effectiveness, medications for opioid use disorder (MOUD) are often stigmatized. Even healthcare professionals working in substance use treatment may describe these medications as undesirable forms of medical intervention. Many argue they prefer an “abstinence-based” approach to treatment, free of pharmacotherapy. Nonetheless, between the three evidence-based U.S. treatment medications, which include methadone, buprenorphine, and naltrexone, there are signs of variation in how stigma operates toward each intervention. Using in-depth semi-structured interviews with 59 U.S. treatment professionals based in New Mexico and Texas, this study shows the drivers of “intervention stigma,” or stigma toward the use or provision of a medical treatment or procedure, toward each type of MOUD. Participants describe a stigma hierarchy framing naltrexone as most acceptable, followed by buprenorphine, and finally methadone, which inverts the relative clinical efficacy of each medication. Pharmacological and regulatory differences, prioritization of psychosocial interventions, suspicion regarding profit motives, and the value placed on their personal experiences as former clients in treatment in tandem with professional anecdote over scientific evidence, contribute to different levels of stigma towards each of the three medications. Acknowledgement of the deleterious effects of stigma toward people who use drugs is growing, including attention to how MOUD is also subject to stigma. This study provides detailed descriptions of the multiple drivers of stigma toward different treatment medications that may serve as key leverage points for future destigmatization efforts.

IntroductionWorking from home during the COVID-19 pandemic has been associated both with physical inactivity and musculoskeletal pain. However, it has not been examined whether physical activity and sedentary behavior are underlying mechanisms in the association between working from home and musculoskeletal pain. Therefore, we examined their mediating role in this association.MethodsData were used from 24 questionnaire rounds of the Lifelines COVID-19 cohort (March 2020–January 2022). Longitudinal information on work situation (location, home, hybrid), physical activity, sedentary behavior, and musculoskeletal pain was collected among 28,586 workers. Analysis of physical activity/sedentary behavior as mediators of the association between working from home and musculoskeletal pain was performed using multilevel structural equation modeling.ResultsHome workers more often had pain in the upper back [odds ratio (OR) = 1.17, 95%-confidence interval (CI) = 1.02–1.34] and arm, neck, and/or shoulder (ANS) (OR = 1.32, 95%-CI = 1.19–1.47) than location workers. Furthermore, home workers were more often sedentary for >9 h per work day than location workers (OR = 2.82, 95%-CI = 2.56–3.09), and being more sedentary was associated with musculoskeletal pain (upper back: OR = 1.17, 95%-CI = 1.06–1.30; ANS: OR = 1.25, 95%-CI = 1.16–1.34). Corresponding indirect effects were OR = 1.18 (95%-CI = 1.04–1.33) and OR = 1.26 (95%-CI = 1.12–1.35). No indirect effect was found for physical activity. Similar indirect effects were observed for hybrid workers.ConclusionHome and hybrid workers were more likely to have pain in the upper musculoskeletal system during the COVID-19 pandemic than location workers, which was partly mediated by increased sedentary behavior, but not by reduced physical activity. Measures to reduce sedentary time in home workers may contribute to preventing musculoskeletal pain.

Neuronal regeneration in the central nervous system (CNS) is an important field of research with relevance to all types of neuronal injuries, including neurodegenerative diseases. The glial scar is a result of the astrocyte response to CNS injury. It is made up of many components creating a complex environment in which astrocytes play various key roles. The glial scar is heterogeneous, diverse and its composition depends upon the injury type and location. The heterogeneity of the glial scar observed in different situations of CNS damage and the consequent implications for axon regeneration have not been reviewed in depth. The gap in this knowledge will be addressed in this review which will also focus on our current understanding of central axonal regeneration and the molecular mechanisms involved. The multifactorial context of CNS regeneration is discussed, and we review newly identified roles for components previously thought to solely play an inhibitory role in central regeneration: astrocytes and p75NTR and discuss their potential and relevance for deciding therapeutic interventions. The article ends with a comprehensive review of promising new therapeutic targets identified for axonal regeneration in CNS and a discussion of novel ways of looking at therapeutic interventions for several brain diseases and injuries.

Abstract Background Little is known about the relationship between shift work and perceived health, including potential underlying mechanisms such as unhealthy behaviors. The aim of this study was to investigate whether unhealthy behaviors mediate the relationship between shift work and perceived mental and physical health, taking into account potential differences by level of education. Methods Data from 1633 workers participating in the Doetinchem Cohort Study during 1995–2016 were used. Being engaged in shift work was determined at 1 year preceding the assessment of health behaviors. Mental and physical health were assessed after 5 years of follow-up by the 5-item Mental Health Inventory and the physical functioning scale of the 36-item Short Form Health Survey. Smoking, physical inactivity, alcohol consumption, and overweight were considered as potential mediators and education was treated as moderator. Moderated mediation analyses using generalized estimated equations were performed. Results Shift work was not statistically significantly related to either mental or physical health. Despite this, statistically significant mediation effects of smoking (Beta − 0.09; 95% Confidence Interval − 0.20 - -0.01, respectively B -0.09; 95%CI -0.21 - -0.01) and physical inactivity (B 0.11; 95%CI 0.03–0.23, respectively B 0.08; 95%CI 0.01–0.18) were found in the relationship between shift work and mental or physical health. Direct and indirect effects outweighed each other in the relationship between shift work and mental health, since the direction of these effects was opposite. The relationship between shift work, unhealthy behavior, and health was not different by educational level. Conclusion Shift workers did not report lower mental or physical health than non-shift workers. Though mediation effects of unhealthy behavior were observed in the relationship between shift work and perceived health, these small effects had minor public health relevance.

Abstract Background This study explores whether a sex difference in sensitivity to (strength of the association) and/or in exposure to (prevalence) determinants of gait speed contributes to the observed lower gait speed among older women compared to men. Methods Data from the Longitudinal Aging Study Amsterdam (LASA) were used. In total 2407 men and women aged 55–81 years were included, with baseline measurements in 1992/2002 and follow-up measurements every 3–4 years for 15/25 years. Multivariable mixed model analysis was used to investigate sex differences in sensitivity (interaction term with sex) and in exposure to (change of the sex difference when adjusted) socio-demographic, lifestyle, social and health determinants of gait speed. Results Women had a 0.054 m/s (95 % CI: 0.076 − 0.033, adjusted for height and age) lower mean gait speed compared to men. In general, men and women had similar determinants of gait speed. However, higher BMI and lower physical activity were more strongly associated with lower gait speed in women compared to men (i.e. higher sensitivity). More often having a lower educational level, living alone and having more chronic diseases, pain and depressive symptoms among women compared to men also contributed to observed lower gait speed in women (i.e. higher exposure). In contrast, men more often being a smoker, having a lower physical activity and a smaller personal network size compared to women contributed to a lower gait speed among men (i.e. higher exposure). Conclusions Both a higher sensitivity and higher exposure to determinants of gait speed among women compared to men contributes to the observed lower gait speed among older women. The identified (modifiable) contributing factors should be taken into account when developing prevention and/or treatment strategies aimed to enhance healthy physical aging. This might require a sex-specific approach in both research and clinical practice, which is currently often lacking.

Abstract New glass compositions containing high concentrations of Tb3+ ions were developed aiming at the production of magneto-optical (MO) fibers. This work reports on the structural and MO properties of a new glass composition based on (100 − x)(41GeO2–25B2O3–4Al2O3–10Na2O–20BaO) − xTb4O7. Morphological analysis (HR-TEM) of the sample with the highest concentration of Tb3+ ions confirmed the homogeneous distribution of Tb3+ ions and the absence of nanoclusters. All the samples presented excellent thermal stability against crystallization (ΔT > 100 °C). An optical fiber was manufactured by a fiber drawing process. The UV–Vis spectra of the glasses showed Tb3+ electronic transitions and optical windows varying from 0.4 to 1.6 μm. The magneto-optical properties and the paramagnetic behaviors of the glasses were investigated using Faraday rotation experiments. The Verdet constant (VB) values were calculated at 500, 650, 880, 1050, 1330, and 1550 nm. The maximum VB values obtained at 650 and 1550 nm for the glass with x = 18 mol% were -128 and − 17.6 rad T−1 m−1, respectively. The VB values at 500 and 1550 nm for the optical fiber containing 8 mol% of Tb4O7 were − 110.2 and − 9.5 rad T−1 m−1, respectively, while the optical loss at around 880 nm was 6.4 dB m−1.

OBJECTIVE: Using a novel mediation method that presents unbiased results even in the presence of exposure–mediator interactions, this study estimated the extent to which working conditions and health behaviors contribute to educational inequalities in self-rated health in the workforce. METHODS: Respondents of the longitudinal Survey of Health, Ageing, and Retirement in Europe (SHARE) in 16 countries were selected, aged 50–64 years, in paid employment at baseline and with information on education and self-rated health (N=15 028). Education, health behaviors [including body mass index (BMI)] and working conditions were measured at baseline and self-rated health at baseline and two-year follow-up. Causal mediation analysis with inverse odds weighting was used to estimate the total effect of education on self-rated health, decomposed into a natural direct effect (NDE) and natural indirect effect (NIE). RESULTS: Lower educated workers were more likely to perceive their health as poor than higher educated workers [relative risk (RR) 1.48, 95% confidence interval (CI) 1.37–1.60]. They were also more likely to have unfavorable working conditions and unhealthy behaviors, except for alcohol consumption. When all working conditions were included, the remaining NDE was RR 1.30 (95% CI 1.15–1.44). When BMI and health behaviors were included, the remaining NDE was RR 1.40 (95% CI 1.27–1.54). Working conditions explained 38% and health behaviors and BMI explained 16% of educational inequalities in health. Including all mediators explained 64% of educational inequalities in self-rated health. CONCLUSIONS: Working conditions and health behaviors explain over half of the educational inequalities in self-rated health. To reduce health inequalities, improving working conditions seems to be more important than introducing health promotion programs in the workforce.

New magnetic glass compositions, based on system (100-x) (60GeO2 -25B2O3 –4Al2O3 -10Na2O–1PbO) – xTb4O7 were prepared by the melt-quenching technique. The materials were systematically characterized with respect to their structural (XRD, EXAFS, XANES and Raman), thermal (DSC), morphological, optical, magnetic and magneto-optical (MO) properties. Morphological and magnetic measurements show the homogeneous distribution of the Tb3+ ions as the magnetic susceptibility increases proportionally to the rare earth content. The optical window covers the range from 0.5 up to 1.6 μm, limited by absorption due to 4f8 -> 4f7 5 d 1and 4f8 -> 4f8 transitions of Tb3+. XANES and EXAFS measurements at the Ge–K edge show that the germanium atoms are always tetrahedrally coordinated, while results at the Tb-LIII edge shows terbium atoms to be eight-coordinated and exclusively present in the oxidation state 3+ independently of the glass composition. MO measurements show that the Verdet constant (VB) increases linearly with the Tb3+ concentration reaching the value of −83.9 rad T−1m−1 at 650 nm for glass containing 8 mol% of Tb4O7. Good thermal stability (ΔT = 200 °C) allowed obtaining an optical fiber of a glass containing 4 mol% of Tb4O7 featuring VB values of −37.9, −23.6 and −5.89 rad T−1 m−1, at 500, 650 and 1550 nm, respectively.

Alzheimer's disease (AD) is the most common form of dementia, which is neuropathologically characterized by extracellular senile plaques containing amyloid-β and intracellular neurofibrillary tangles composed of hyperphosphorylated tau protein. Previous studies have suggested a role for septin (SEPTIN) protein family members in AD-associated cellular processes. Here, we elucidated the potential role of presynaptic SEPTIN5 protein and its post-translational modifications in the molecular pathogenesis of AD. RNA and protein levels of SEPTIN5 showed a significant decrease in human temporal cortex in relation to the increasing degree of AD-related neurofibrillary pathology. Conversely, an increase in the phosphorylation of the functionally relevant SEPTIN5 phosphorylation site S327 was observed already in the early phases of AD-related neurofibrillary pathology, but not in the cerebrospinal fluid of individuals fulfilling the criteria for mild cognitive impairment due to AD. According to the mechanistic assessments, a link between SEPTIN5 S327 phosphorylation status and the effects of SEPTIN5 on amyloid precursor protein processing and markers of autophagy was discovered in mouse primary cortical neurons transduced with lentiviral constructs encoding wild type SEPTIN5 or SEPTIN5 phosphomutants (S327A and S327D). C57BL/6 J mice intrahippocampally injected with lentiviral wild type SEPTIN5 or phosphomutant constructs did not show changes in cognitive performance after five to six weeks from the start of injections. However, SEPTIN5 S327 phosphorylation status was linked to changes in short-term synaptic plasticity ex vivo at the CA3-CA1 synapse. Collectively, these data suggest that SEPTIN5 and its S327 phosphorylation status play a pivotal role in several cellular processes relevant for AD.

This study aims to identify the temporal variation and the spatial dependence structure of the hospitalization rate for falls in the elderly residing in Brazil in the period between 2010 and 2021. This ecological study employs secondary data from the Brazilian Ministry of Health about the fall-related hospitalization of people aged 60 years old and over. A time-series analysis was carried out, employing the joinpoint model. For the spatial analysis, the Moran autocorrelation technique was employed. In Brazil, between 2010 and 2021, there were 1,270,341 hospitalizations for falls recorded among the elderly in the Brazilian Hospitalization System. There was a continuous upward trend between 2010 and 2019 for all age groups, female and male, and all Brazilian regions. The trend stabilized between 2019 to 2021. The North and Northeast regions had faster upward trends among all Brazilian regions, and there was also a faster upward trend among women compared to men. A high-high pattern in hospitalization incidence was noticed from 2011 to 2019 in the states of São Paulo, Minhas Gerais, Paraná, and Mato Grosso do Sul. The results of this study provide subsidies for Brazilian health authorities to implement more efficient public policies to improve the quality of life of elderly people.

Undergraduate teacher candidates participated in a faculty-led, short-term study abroad to Reggio Emilia in Italy to further their knowledge about the educational philosophy used within the school setting and for the purpose of examining how philosophy might be applied in school settings in the United States. The faculty-led, short-term study abroad included two days of travel, two days of sightseeing, and four days of lecture, atelier experiences, visits to infant-toddler centers and preschools, and encounters with atelieristas, pedagogistas, and Reggio educators. Undergraduate teacher candidates (hereafter referred to as teacher candidates) met three times prior to travel to discuss philosophy and travel arrangements. Each teacher candidate was provided a journal to be used for note-taking and reflections while in Reggio Emilia. Teacher candidates used a thinking routine, Wish-Wonder-Wow, to provide structure for notes and reflections. While in Reggio Emilia, teacher candidates shared reflections nightly in a group format. This study examines 12 teacher candidates' perspectives from these study abroad experiences that occurred over a four-year period from 2016-2019. The researchers include a faculty member leading the study abroad for two of the years as well as another faculty member who has experienced the Reggio Emilia setting in Italy. At the time of the study, the teacher candidates who participated in the study abroad were student teaching or had graduated and were teachers with one to three of years of experience. Their responses to a survey (n=12) and focus group interviews (n=7) are viewed through the lenses of the four factors in Lundy's (2007) model of including children in decision-making and focused on space, voice, audience, and influence in reference to Article 12 of the United Nations Convention on the Rights of the Child (United Nations General Assembly, 2002). Through this study, the researchers learned teacher candidates' thinking and philosophical struggles with classroom implementation. Survey and focus group data indicated that short-term study abroad enhanced teacher candidates' understanding and appreciation of the Reggio Emilia Approach to Education. However, teacher candidates felt a lack of support and guidance as a new teacher in being able to incorporate the philosophical beliefs into their classrooms. This proved challenging to them, and the participants lacked confidence in implementing change within a school setting.

Amyotrophic lateral sclerosis (ALS) is characterized by the progressive degeneration of corticospinal tract motor neurons. Previous studies showed that adenosine-mediated neuromodulation is disturbed in ALS and that vascular endothelial growth factor (VEGF) has a neuroprotective function in ALS mouse models. We evaluated how adenosine (A1R and A2AR) and VEGF (VEGFA, VEGFB, VEGFR-1 and VEGFR-2) system markers are altered in the cortex and spinal cord of pre-symptomatic and symptomatic SOD1G93A mice. We then assessed if/how chronic treatment of SOD1G93A mice with a widely consumed adenosine receptor antagonist, caffeine, modulates VEGF system and/or the levels of Brain-derived Neurotrophic Factor (BDNF), known to be under control of A2AR. We found out decreases in A1R and increases in A2AR levels even before disease onset. Concerning the VEGF system, we detected increases of VEGFB and VEGFR-2 levels in the spinal cord at pre-symptomatic stage, which reverses at the symptomatic stage, and decreases of VEGFA levels in the cortex, in very late disease states. Chronic treatment with caffeine rescued cortical A1R levels in SOD1G93A mice, bringing them to control levels, while rendering VEGF signaling nearly unaffected. In contrast, BDNF levels were significantly affected in SOD1G93A mice treated with caffeine, being decreased in the cortex and increased in spinal the cord. Altogether, these findings suggest an early dysfunction of the adenosinergic system in ALS and highlights the possibility that the negative influence of caffeine previously reported in ALS animal models results from interference with BDNF rather than with the VEGF signaling molecules.

This paper presents the influence of the presence of a modified organoclay, Cloisite® 20A (MMTA) on the structural and drug release properties of ureasil organic–inorganic hybrid. Sol–gel process was used to prepare the hybrid nanocomposites containing sodium diclofenac (DCF) at 5% wt. The effect of the amount of MMTA incorporated into the ureasil hybrid matrix was evaluated and characterized in depth by different techniques such as X-ray diffraction (XRD), small angle X-ray scattering (SAXS), differential scanning calorimetry (DSC), and swelling properties. The influence of MMTA on ureasil nanocomposites release profile was evaluated by in situ UV–vis. The diffraction patterns of the UPEO–MMTA nanocomposites showed a synergistic contribution effect that led to an intensity increase and narrowed the diffraction peaks, evidencing a crystallite PEO growth as a function of the modified nanoclay content. The interactions between polyether chains and the hydrogenated tallow of MMTA led to an easy intercalation process, as observed in UPEO–MMTA nanocomposites containing low (1% wt) or high (20% wt) nanoclay content. The waterway (channels) created in UPEO–MMTA nanocomposites contributed to a free volume increase in the swollen network compared to UPEO without MMTA. The hypothesis of the channels created after intercalation of the PEO phase in the interlayer of MMTA containing organoammonium ions corroborates with the XRD results, swelling studies by SAXS, and release assays. Furthermore, when these clay particles were dispersed in the polymeric matrix by an intercalation process, water uptake improvement was observed, with an increased amount of DCF release. The design of ureasil-MMTA nanocomposites containing modified nanoclay endows them with tunable properties; for example, swelling degree followed by amount of controlled drug release, opening the way for more versatile biomedical applications.

OBJECTIVE: This systematic review aimed to provide an overview of the effectiveness of health promotion interventions at the workplace on physical and mental health outcomes related to chronic diseases. METHODS: A search for reviews published between 2009 and 2018 was performed in electronic databases. References of the included reviews were checked for additional reviews. Workplace health promotion interventions were included if they studied metabolic risk factors as important predictors of type 2 diabetes mellitus (T2DM) and cardiovascular diseases (CVD) or if they studied mental or musculoskeletal health outcomes. Review quality was assessed using the AMSTAR checklist. RESULTS: Of the 23 reviews included, 9 were of high quality. For weight-related outcomes, there was strong evidence for favorable effects of workplace interventions, especially for interventions targeting physical activity and/or diet. For the remaining metabolic risk factors, there was no evidence for a positive effect of workplace health promotion interventions due to the absence of high quality reviews and mixed conclusions between the reviews. There was also strong evidence for a positive, small effect on the prevention of mental health disorders of workplace psychological interventions, especially those that use e- health and cognitive behavioral therapy techniques. Furthermore, strong evidence was found for the prevention of musculoskeletal disorders through workplace interventions, especially resistance exercise training. CONCLUSIONS: This review found evidence for the effectiveness of workplace interventions on the prevention of weight-related outcomes as well as mental health and musculoskeletal disorders. Future research is however needed on the factors that contribute to the successful implementation of an intervention.

Reactive astrocytes in Amyotrophic Lateral Sclerosis (ALS) change their molecular expression pattern and release toxic factors that contribute to neurodegeneration and microglial activation. We and others identified a dysregulated inflammatory miRNA profile in ALS patients and in mice models suggesting that they represent potential targets for therapeutic intervention. Such cellular miRNAs are known to be released into the secretome and to be carried by small extracellular vesicles (sEVs), which may be harmful to recipient cells. Thus, ALS astrocyte secretome may disrupt cell homeostasis and impact on ALS pathogenesis. Previously, we identified a specific aberrant signature in the cortical brain of symptomatic SOD1-G93A (mSOD1) mice, as well as in astrocytes isolated from the same region of 7-day-old mSOD1 mice, with upregulated S100B/HMGB1/Cx43/vimentin and downregulated GFAP. The presence of downregulated miR-146a on both cases suggests that it can be a promising target for modulation in ALS. Here, we upregulated miR-146a with pre-miR-146a, and tested glycoursodeoxycholic acid (GUDCA) and dipeptidyl vinyl sulfone (VS) for their immunoregulatory properties. VS was more effective in restoring astrocytic miR-146a, GFAP, S100B, HMGB1, Cx43, and vimentin levels than GUDCA, which only recovered Cx43 and vimentin mRNA. The miR-146a inhibitor generated typical ALS aberrancies in wild type astrocytes that were abolished by VS. Similarly, pre-miR-146a transfection into the mSOD1 astrocytes abrogated aberrant markers and intracellular Ca2+ overload. Such treatment counteracted miR-146a depletion in sEVs and led to secretome-mediated miR-146a enhancement in NSC-34-motor neurons (MNs) and N9-microglia. Secretome from mSOD1 astrocytes increased early/late apoptosis and FGFR3 mRNA in MNs and microglia, but not when derived from pre-miR-146a or VS-treated cells. These last strategies prevented the impairment of axonal transport and synaptic dynamics by the pathological secretome, while also averted microglia activation through either secretome, or their isolated sEVs. Proteomic analysis of the target cells indicated that pre-miR-146a regulates mitochondria and inflammation via paracrine signaling. We demonstrate that replenishment of miR-146a in mSOD1 cortical astrocytes with pre-miR-146a or by VS abrogates their phenotypic aberrancies and paracrine deleterious consequences to MNs and microglia. These results propose miR-146a as a new causal and emerging therapeutic target for astrocyte pathogenic processes in ALS.

Engineering brain organoids from human induced pluripotent stem cells (hiPSCs) is a powerful tool for modeling brain development and neurological disorders. Rett syndrome (RTT), a rare neurodevelopmental disorder, can greatly benefit from this technology, since it affects multiple neuronal subtypes in forebrain sub-regions. We have established dorsal and ventral forebrain organoids from control and RTT patient-specific hiPSCs recapitulating 3D organization and functional network complexity. Our data revealed a premature development of the deep-cortical layer, associated to the formation of TBR1 and CTIP2 neurons, and a lower expression of neural progenitor/proliferative cells in female RTT dorsal organoids. Moreover, calcium imaging and electrophysiology analysis demonstrated functional defects of RTT neurons. Additionally, assembly of RTT dorsal and ventral organoids revealed impairments of interneuron’s migration. Overall, our models provide a better understanding of RTT during early stages of neural development, demonstrating a great potential for personalized diagnosis and drug screening.

Phytocannabinoids are psychotropic substances ofcannabis with the ability to bind endocannabinoid (eCB) receptors that regulate synaptic activity in the central nervous system (CNS). Synthetic cannabinoids (SCs) are synthetic analogs of Δ9-tetrahydrocannabinol (Δ9-THC), the psychotropic compound of cannabis, acting as agonists of eCB receptor CB1. SC is an easily available and popular alternative to cannabis, and their molecular structure is always changing, increasing the hazard for the general population. The popularity of cannabis and its derivatives may lead, and often does, to a child’s exposure to cannabis both in utero and through breastfeeding by a drug-consuming mother. Prenatal exposure to cannabis has been associated with an altered rate of mental development and significant changes in nervous system functioning. However, the understanding of mechanisms of its action on developing the human CNS is still lacking. We investigated the effect of continuous exposure to cannabinoids on developing human neurons, mimicking the prenatal exposure by drug-consuming mother. Two human induced pluripotent stem cells (hiPSC) lines were induced to differentiate into neuronal cells and exposed for 37 days to cannabidiol (CBD), Δ9-THC, and two SCs, THJ-018 and EG-018. Both Δ9-THC and SC, at 10 μM, promote precocious neuronal and glial differentiation, while CBD at the same concentration is neurotoxic. Neurons exposed to Δ9-THC and SC show abnormal functioning of voltage-gated calcium channels when stimulated by extracellular potassium. In sum, all studied substances have a profound impact on the developing neurons, highlighting the importance of thorough research on the impact of prenatal exposure to natural and SC.

Excitotoxicity is a phenomenon that describes the toxic actions of excitatory neurotransmitters, primarily glutamate, where the exacerbated or prolonged activation of glutamate receptors starts a cascade of neurotoxicity that ultimately leads to the loss of neuronal function and cell death. In this process, the shift between normal physiological function and excitotoxicity is largely controlled by astrocytes since they can control the levels of glutamate on the synaptic cleft. This control is achieved through glutamate clearance from the synaptic cleft and its underlying recycling through the glutamate-glutamine cycle. The molecular mechanism that triggers excitotoxicity involves alterations in glutamate and calcium metabolism, dysfunction of glutamate transporters, and malfunction of glutamate receptors, particularly N-methyl-D-aspartic acid receptors (NMDAR). On the other hand, excitotoxicity can be regarded as a consequence of other cellular phenomena, such as mitochondrial dysfunction, physical neuronal damage, and oxidative stress. Regardless, it is known that the excessive activation of NMDAR results in the sustained influx of calcium into neurons and leads to several deleterious consequences, including mitochondrial dysfunction, reactive oxygen species (ROS) overproduction, impairment of calcium buffering, the release of pro-apoptotic factors, among others, that inevitably contribute to neuronal loss. A large body of evidence implicates NMDAR-mediated excitotoxicity as a central mechanism in the pathogenesis of many neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), Alzheimer’s disease (AD), and epilepsy. In this review article, we explore different causes and consequences of excitotoxicity, discuss the involvement of NMDAR-mediated excitotoxicity and its downstream effects on several neurodegenerative disorders, and identify possible strategies to study new aspects of these diseases that may lead to the discovery of new therapeutic approaches. With the understanding that excitotoxicity is a common denominator in neurodegenerative diseases and other disorders, a new perspective on therapy can be considered, where the targets are not specific symptoms, but the underlying cellular phenomena of the disease.

The cerebellum plays a critical role in all vertebrates, and many neurological disorders are associated with cerebellum dysfunction. A major limitation in cerebellar research has been the lack of adequate disease models. As an alternative to animal models, cerebellar neurons differentiated from pluripotent stem cells have been used. However, previous studies only produced limited amounts of Purkinje cells. Moreover, in vitro generation of Purkinje cells required co-culture systems, which may introduce unknown components to the system. Here we describe a novel differentiation strategy that uses defined medium to generate Purkinje cells, granule cells, interneurons, and deep cerebellar nuclei projection neurons, that self-formed and differentiated into electrically active cells. Using a defined basal medium optimized for neuronal cell culture, we successfully promoted the differentiation of cerebellar precursors without the need for co-culturing. We anticipate that our findings may help developing better models for the study of cerebellar dysfunctions, while providing an advance toward the development of autologous replacement strategies for treating cerebellar degenerative diseases.

Abstract Background Applying very high gravity (VHG) fermentation conditions to the sugarcane juice (SCJ) bioethanol industry would improve its environmental and economic sustainability without the need for major infrastructure changes or investments. It could enable a decrease in the consumption of biological and natural resources (cane/land, water and energy) while maintaining acceptable production parameters. The present study attempts to demonstrate and characterise an effective industrially relevant SCJ-VHG fermentation process. Results An industry-like SCJ-VHG bioethanol production process with 30 and 35 °Bx broth was employed to investigate the effects of both the yeast strain used and nitrogen source supplementation on process yield, process productivity, biomass viability, glycerol concentration and retention-associated gene expression. Process performance was shown to be variably affected by the different process conditions investigated. Highest process efficiency, with a 17% (w/v) ethanol yield and only 0.2% (w/v) sugar remaining unfermented, was observed with the Saccharomyces cerevisiae industrial strain CAT-1 in 30 °Bx broth with urea supplementation. In addition, efficient retention of glycerol by the yeast strain was identified as a requisite for better fermentation and was consistent with a higher expression of glycerol permease STL1 and channel FPS1. Urea was shown to promote the deregulation of STL1 expression, overcoming glucose repression. The consistency between Fps1-mediated ethanol secretion and ethanol in the extracellular media reinforces previous suggestions that ethanol might exit the cell through the Fps1 channel. Conclusions This work brings solid evidence in favour of the utilisation of VHG conditions in SCJ fermentations, bringing it a step closer to industrial application. SCJ concentrated up to 30 °Bx maintains industrially relevant ethanol production yield and productivity, provided the broth is supplemented with a suitable nitrogen source and an appropriate industrial bioethanol-producing yeast strain is used. In addition, the work contributes to a better understanding of the VHG-SCJ process and the variable effects of process parameters on process efficiency and yeast strain response.

Metal bioavailability is frequently limited by sequestering agents which makes them inaccessible to cells. Here the authors show that cells can increase Mg2+ uptake via fluid phase endocytosis and accumulate this metal in their vacuole loaded with polyphosphate, and later can be exported to the cytosol.

Abstract Background Accumulation of problems in physical, psychological, cognitive, or social functioning is characteristic for frail individuals. Using a four-domain approach of frailty, this study explored how sociodemographic and lifestyle factors, life events and health are associated with frailty. Methods The study sample included 4019 men and women (aged 40–81 years) examined during the fifth round (2008–2012) of the Doetinchem Cohort Study. Four domains of frailty were considered: physical (≥4 of 8 criteria: unintentional weight loss, exhaustion, strength, perceived health, walking, balance, hearing and vision impairments), psychological (2 criteria: depressive symptoms, mental health), cognitive (

The use of immersive simulation as a pedagogical tool has great potential for making a significant impact on student learning in higher education. In this study, the effect of immersive simulation was evaluated for a cohort of undergraduate special education majors. The investigation aimed to determine whether facilitating an immersive co-planning simulation would have an impact on targeted collaboration skills and whether vicarious observational learning would occur for students who observed the simulation. Pre-service teachers in special education were evaluated by their peers on their ability to demonstrate knowledge of (1) co-teaching and co-planning, (2) professional communication, and (3) supports for students with disabilities. The results indicate that they did a better job of facilitating a co-planning session after having first practiced doing so via immersive simulation during a previous class session. It was also discovered that vicarious observational learning during immersive simulation positively affected performance.