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Negative cognitions play a key role in the development and maintenance of depression. To reduce depressive symptoms, most interventions either encourage adolescents to change negative cognitions, theorizing that the presence of negative cognitions underlies depression, or to acknowledge negative cognitions, theorizing that one's reaction to negative cognitions underlies depression. We compared these two therapeutic strategies in a multilevel meta-analysis of the effects of changing versus acknowledging cognitions on adolescent depression. We searched three databases in June 2022 and identified 104 randomized controlled trials (335 effect sizes). The sample comprised 27,978 adolescents (sample mean age 14-18 years) with all levels of depressive symptoms (M[subscript age] = 15.6 years; 63% female; 65% ethnic majority). The overall effect of interventions on depression was small (d = 0.21, p < 0.001). We found no evidence that either strategy was superior to the other. Strategies to acknowledge (d = 0.23, p = 0.016) or change cognitions (d = 0.20, p < 0.001) both reduced adolescent depression. Our findings suggest, though based on self-reported outcomes, that both strategies are effective in reducing adolescent depression, which allows for flexibility for clinicians and patients. The next step to further understand these strategies is to scrutinize the relative effects of single versus combined approaches to change and acknowledge negative cognitions.

To writing anything on a keyboard at all requires us to know first what to type, then to activate motor programmes for finger movements, and execute these. An interruption in the information flow at any of these stages leads to disfluencies. To capture this combination of fluent typing and typing hesitations, researchers calculate different measures from keystroke-latency data--such as mean inter-keystroke interval and pause frequencies. There are two fundamental problems with this: first, summary statistics ignore important information in the data and frequently result in biased estimates; second, pauses and pause-related measures are defined using threshold values which are, in principle, arbitrary. We implemented a series of Bayesian models that aimed to address both issues while providing reliable estimates for individual typing speed and statistically inferred process disfluencies. We tested these models on a random sample of 250 copy-task recordings. Our results illustrate that we can model copy typing as a mixture process of fluent and disfluent key transitions. We conclude that mixture models (1) map onto the information cascade that generate keystrokes, and (2) provide a principled approach to detect disfluencies in keyboard typing.

Abstract Background The access to and uptake of evidence-based behavioral parent training for children with behavioral difficulties (i.e., oppositional, defiant, aggressive, hyperactive, impulsive, and inattentive behavior) are currently limited because of a scarcity of certified therapists and long waiting lists. These problems are in part due to the long and sometimes perceived as rigid nature of most evidence-based programs and result in few families starting behavioral parent training and high dropout rates. Brief and individually tailored parenting interventions may reduce these problems and make behavioral parent training more accessible. This protocol paper describes a two-arm, multi-center, randomized controlled trial on the short- and longer-term effectiveness and cost-effectiveness of a brief, individually tailored behavioral parent training program for children with behavioral difficulties. Methods Parents of children aged 2–12 years referred to a child mental healthcare center are randomized to (i) three sessions of behavioral parent training with optional booster sessions or (ii) care as usual. To evaluate effectiveness, our primary outcome is the mean severity of five daily ratings by parents of four selected behavioral difficulties. Secondary outcomes include measures of parent and child behavior, well-being, and parent–child interaction. We explore whether child and parent characteristics moderate intervention effects. To evaluate cost-effectiveness, the use and costs of mental healthcare and utilities are measured. Finally, parents’ and therapists’ satisfaction with the brief program are explored. Measurements take place at baseline (T0), one week after the brief parent training, or eight weeks after baseline (in case of care as usual) (T1), and six months (T2) and twelve months (T3) after T1. Discussion The results of this trial could have meaningful societal implications for children with behavioral difficulties and their parents. If we find the brief behavioral parent training to be more (cost-)effective than care as usual, it could be used in clinical practice to make parent training more accessible. Trial registration The trial is prospectively registered at ClinicalTrials.gov (NCT05591820) on October 24th, 2022 and updated throughout the trial.

Abstract Osteoarthritis (OA) is a multifactorial degenerative joint disease of which the underlying mechanisms are yet to be fully understood. At the molecular level, multiple factors including altered signaling pathways, epigenetics, metabolic imbalance, extracellular matrix degradation, production of matrix metalloproteinases, and inflammatory cytokines, are known to play a detrimental role in OA. However, these factors do not initiate OA, but are mediators or consequences of the disease, while many other factors causing the etiology of OA are still unknown. Here, it is revealed that microenvironmental osmolarity can induce and reverse osteoarthritis‐related behavior of chondrocytes via altered intracellular molecular crowding, which represents a previously unknown mechanism underlying OA pathophysiology. Decreased intracellular crowding is associated with increased sensitivity to proinflammatory triggers and decreased responsiveness to anabolic stimuli. OA‐induced lowered intracellular molecular crowding could be renormalized via exposure to higher extracellular osmolarity such as those found in healthy joints, which reverse OA chondrocyte's sensitivity to catabolic stimuli as well as its glycolytic metabolism.

Abstract Organoids are engineered 3D miniature tissues that are defined by their organ-like structures, which drive a fundamental understanding of human development. However, current organoid generation methods are associated with low production throughputs and poor control over size and function including due to organoid merging, which limits their clinical and industrial translation. Here, we present a microfluidic platform for the mass production of lumenogenic embryoid bodies and functional cardiospheres. Specifically, we apply triple-jet in-air microfluidics for the ultra-high-throughput generation of hollow, thin-shelled, hydrogel microcapsules that can act as spheroid-forming bioreactors in a cytocompatible, oil-free, surfactant-free, and size-controlled manner. Uniquely, we show that microcapsules generated by in-air microfluidics provide a lumenogenic microenvironment with near 100% efficient cavitation of spheroids. We demonstrate that upon chemical stimulation, human pluripotent stem cell-derived spheroids undergo cardiomyogenic differentiation, effectively resulting in the mass production of homogeneous and functional cardiospheres that are responsive to external electrical stimulation. These findings drive clinical and industrial adaption of stem cell technology in tissue engineering and drug testing.

Highly pathogenic avian influenza viruses (HPAIVs) cause severe hemorrhagic disease in terrestrial poultry and are a threat to the poultry industry, wild life, and human health. HPAIVs arise from low pathogenic avian influenza viruses (LPAIVs), which circulate in wild aquatic birds. HPAIV emergence is thought to occur in poultry and not wild aquatic birds, but the reason for this species-restriction is not known. We hypothesized that, due to species-specific tropism and replication, intrahost HPAIV selection is favored in poultry and disfavored in wild aquatic birds. We tested this hypothesis by co-inoculating chickens, representative of poultry, and ducks, representative of wild aquatic birds, with a mixture of H7N7 HPAIV and LPAIV, mimicking HPAIV emergence in an experimental setting. Virus selection was monitored in swabs and tissues by RT-qPCR and immunostaining of differential N-terminal epitope tags that were added to the hemagglutinin protein. HPAIV was selected in four of six co-inoculated chickens, whereas LPAIV remained the major population in co-inoculated ducks on the long-term, despite detection of infectious HPAIV in tissues at early time points. Collectively, our data support the hypothesis that HPAIVs are more likely to be selected at the intrahost level in poultry than in wild aquatic birds and point towards species-specific differences in HPAIV and LPAIV tropism and replication levels as possible explanations.

Protein-protein interaction (PPI) networks are fundamental for cellular processes. Small-molecule PPI enhancers have been shown to be powerful tools to fundamentally study PPIs and as starting points for potential new therapeutics. Yet, systematic approaches for their discovery are not widely available, and the design prerequisites of "molecular glues" are poorly understood. Covalent fragment-based screening can identify chemical starting points for these enhancers at specific sites in PPI interfaces. We recently reported a mass spectrometry-based disulfide-trapping (tethering) approach for a cysteine residue in the hub protein 14-3-3, an important regulator of phosphorylated client proteins. Here, we invert the strategy and report the development of a functional read-out for systematic identification of PPI enhancers based on fluorescence anisotropy (FA-tethering) with the reactive handle now on a client-derived peptide. Using the DNA-binding domain of the nuclear receptor Estrogen Related Receptor gamma (ERRγ), we target a native cysteine positioned at the 14-3-3 PPI interface and identify several fragments that form a disulfide bond to ERRγ and stabilize the complex up to 5-fold. Crystallography indicates that fragments bind in a pocket comprised of 14-3-3 and the ERRγ phosphopeptide. FA-tethering presents a streamlined methodology to discover molecular glues for protein complexes.

We previously demonstrated that diet supplementation with seaweed Sargassum fusiforme (S. fusiforme) prevented AD-related pathology in a mouse model of Alzheimer’s Disease (AD). Here, we tested a lipid extract of seaweed Himanthalia elongata (H. elongata) and a supercritical fluid (SCF) extract of S. fusiforme that is free of excess inorganic arsenic. Diet supplementation with H. elongata extract prevented cognitive deterioration in APPswePS1ΔE9 mice. Similar trends were observed for the S. fusiforme SCF extract. The cerebral amyloid-β plaque load remained unaffected. However, IHC analysis revealed that both extracts lowered glial markers in the brains of APPswePS1ΔE9 mice. While cerebellar cholesterol concentrations remained unaffected, both extracts increased desmosterol, an endogenous LXR agonist with anti-inflammatory properties. Both extracts increased cholesterol efflux, and particularly, H. elongata extract decreased the production of pro-inflammatory cytokines in LPS-stimulated THP-1-derived macrophages. Additionally, our findings suggest a reduction of AD-associated phosphorylated tau and promotion of early oligodendrocyte differentiation by H. elongata. RNA sequencing on the hippocampus of one-week-treated APPswePS1ΔE9 mice revealed effects of H. elongata on, amongst others, acetylcholine and synaptogenesis signaling pathways. In conclusion, extracts of H. elongata and S. fusiforme show potential to reduce AD-related pathology in APPswePS1ΔE9 mice. Increasing desmosterol concentrations may contribute to these effects by dampening neuroinflammation.

Osteoarthritis (OA) is a degenerative disease of the joints for which no curative treatment exists. Intra-articular injection of stem cells is explored as a regenerative approach, but rapid clearance of cells from the injection site limits the therapeutic outcome. Microencapsulation of mesenchymal stem cells (MSCs) can extend the retention time of MSCs, but the outcomes of the few studies currently performed are conflicting. We hypothesize that the composition of the micromaterial's shell plays a deciding factor in the treatment outcome of intra-articular MSC injection. To this end, we microencapsulate MSCs using droplet microfluidic generators in flow-focus mode using various polymers and polymer concentrations. We demonstrate that polymer composition and concentration potently alter the metabolic activity as well as the secretome of MSCs. Moreover, while microencapsulation consistently prolongs the retention time of MSC injected in rat joints, distinct biodistribution within the joint is demonstrated for the various microgel formulations. Furthermore, intra-articular injections of pristine and microencapsulated MSC in OA rat joints show a strong material-dependent effect on the reduction of cartilage degradation and matrix loss. Collectively, this study highlights that micromaterial composition and concentration are key deciding factors for the therapeutic outcome of intra-articular injections of microencapsulated stem cells to treat degenerative joint diseases.

Abstract Background Behavioural parent training is an evidence‐based intervention for children with attention‐deficit/hyperactivity disorder (ADHD), but little is known about the extent to which initial benefits are maintained. Aims This meta‐analytic review investigated longer‐term (i.e., more than 2 months post‐intervention) child and parental outcomes of behavioural parent training for children with ADHD. Materials & Methods We searched for randomized controlled trials and examined ADHD symptoms, behavioural problems, positive parenting, negative parenting, parenting sense of competence, parent‐child relationship quality, and parental mental health as outcomes. We included 27 studies (31 interventions; 217 effect sizes), used multilevel random‐effects meta‐analyses for between‐ and within‐group comparisons (pre‐intervention to follow‐up and post‐intervention to follow‐up), and explored twelve predictors of change. Results Between pre‐intervention and follow‐up (M = 5.3 months), we found significant small‐to‐moderate between‐group effects of the intervention on ADHD symptoms, behavioural problems, positive parenting, parenting sense of competence and parent‐child relationship quality. Within‐group findings show sustained improvements in the intervention conditions for all outcome domains. There were few significant changes from post‐intervention to follow‐up. Additionally, the large majority of the individual effect sizes indicated sustained outcomes from post‐intervention to follow‐up. There were seven significant predictors of change in child outcomes, including stronger reductions in ADHD symptoms of girls and behaviour problems of younger children. In contrast with some meta‐analyses on short‐term effects, we found no differences between masked and unmasked outcomes on ADHD symptoms at follow‐up. Discussion & Conclusion We conclude that behavioural parent training has longer‐term benefits for children's ADHD symptoms and behavioural problems, and for positive parenting behaviours, parenting sense of competence and quality of the parent‐child relationship.

Living microtissues are used in a multitude of applications as they more closely resemble native tissue physiology, as compared to 2D cultures. Microtissues are typically composed of a combination of cells and materials in varying combinations, which are dictated by the applications’ design requirements. Their applications range wide, from fundamental biological research such as differentiation studies to industrial applications such as cruelty-free meat production. However, their translation to industrial and clinical settings has been hindered due to the lack of scalability of microtissue production techniques. Continuous microfluidic processes provide an opportunity to overcome this limitation as they offer higher throughput production rates as compared to traditional batch techniques, while maintaining reproducible control over microtissue composition and size. In this review, we provide a comprehensive overview of the current approaches to engineer microtissues with a focus on the advantages of, and need for, the use of continuous processes to produce microtissues in large quantities. Finally, an outlook is provided that outlines the required developments to enable large-scale microtissue fabrication using continuous processes.

Abstract In psoriatic arthritis (PsA), predisposing class I HLA alleles, the presence of synovial clonally proliferated CD8 + T cells and autoantibodies all point towards the loss of immune tolerance. However, the key mechanisms that lead to immune dysregulation are not fully understood. In other types of inflammatory arthritis, T regulatory cell (Treg) dysfunction and plasticity at sites of inflammation were suggested to negatively affect peripheral tolerance. We here addressed if Treg variances associate with psoriatic disease. We collected clinical data, sera and peripheral blood mononuclear cells from 13 healthy controls, 21 psoriasis and 21 PsA patients. In addition, we obtained synovial fluid mononuclear cells from 6 PsA patients. We studied characteristics of CD4 + CD25 + CD127loFoxp3 + Tregs by flow cytometry and used ELISA to quantify antibodies against ADAMTSL5, a recently discovered autoantigen in psoriatic disease. In comparison with their circulating counterparts, Tregs from inflamed joints express increased levels of ICOS, CTLA-4 and TIGIT. Furthermore, synovial fluid-derived Tregs have a distinct phenotype, characterized by IL-17A production and upregulation of CD161 and RORγt. We identified a subset of Tregs with intermediate Foxp3 expression as the major cytokine producer. Furthermore, ICOS + Tregs associate with PsA disease activity as measured by PASDAS. Lastly, we observed that presence of the Foxp3int Tregs associates with an increased abundance of anti-ADAMTSL5 autoantibodies. Tregs derived from the inflammatory environment of inflamed PsA joints exhibit a distinct phenotype, which associates with loss of peripheral immune tolerance in psoriatic disease.

Abstract Objective To assess the performance of a multimodal seizure detection device, first tested in adults (sensitivity 86%, PPV 49%), in a pediatric cohort living at home or residential care. Methods In this multicenter, prospective, video‐controlled cohort‐study, nocturnal seizures were detected by heartrate and movement changes in children with epilepsy and intellectual disability. Participants with a history of >1 monthly major motor seizure wore Nightwatch bracelet at night for 3 months. Major seizures were defined as tonic–clonic, generalized tonic >30 s, hyperkinetic, or clusters (>30 min) of short myoclonic or tonic seizures. The video of all events (alarms and nurse diaries) and about 10% of whole nights were reviewed to classify major seizures, and minor or no seizures. Results Twenty‐three participants with focal or generalized epilepsy and nightly motor seizures were evaluated during 1511 nights, with 1710 major seizures. First 1014 nights, 4189 alarms occurred with average of 1.44/h, showing average sensitivity of 79.9% (median 75.4%) with mean PPV of 26.7% (median 11.1%) and false alarm rate of 0.2/hour. Over 90% of false alarms in children was due to heart rate (HR) part of the detection algorithm. To improve this rate, an adaptation was made such that the alarm was only triggered when the wearer was in horizontal position. For the remaining 497 nights, this was tested prospectively, 384 major seizures occurred. This resulted in mean PPV of 55.5% (median 58.1%) and a false alarm rate 0.08/h while maintaining a comparable mean sensitivity of 79.4% (median 93.2%). Significance Seizure detection devices that are used in bed which depend on heartrate and movement show similar sensitivity in children and adults. However, children do show general higher false alarm rate, mostly triggered while awake. By correcting for body position, the false alarms can be limited to a level that comes close to that in adults.

Abstract Interictal 18F‐Fluorodeoxyglucose positron emission tomography (FDG‐PET) is used in the workup for epilepsy surgery when MRI and EEG video monitoring are not conclusive. Timing of FDG‐PET is crucial to avoid the metabolically dynamic (post)ictal state that complicates interpretation, but the exact time window is unclear. We performed a systematic review to provide an evidence‐based recommendation for the minimal time interval between last seizure and FDG‐PET acquisition. We searched PubMed and Embase for articles on the effect of time since last seizure on FDG‐PET outcome. Quality assessment was conducted with the Critical Appraisal Skills Programme Cohort Study Checklist. We identified five studies. Three studies were classified as of low to moderate quality, mainly due to undocumented data or insufficient statistical measurements. Two high‐quality studies included only adults with Temporal Lobe Epilepsy (TLE). The metabolic interictal phase is 24 or 48 hours after the last seizure, depending on seizure type. The recommendation is based on the best available evidence from two small study populations for TLE. If clinically possible, interictal FDG‐PET in adults should be performed at least 24 hours after focal aware seizures and 48 hours after focal impaired awareness and focal to bilateral tonic–clonic seizures.

Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is associated with various neurological complications. Although the mechanism is not fully understood, several studies have shown that neuroinflammation occurs in the acute and post-acute phase. As these studies have predominantly been performed with isolates from 2020, it is unknown if there are differences among SARS-CoV-2 variants in their ability to cause neuroinflammation. Here, we compared the neuroinvasiveness, neurotropism and neurovirulence of the SARS-CoV-2 ancestral strain D614G, the Delta (B.1.617.2) and Omicron BA.1 (B.1.1.529) variants using in vitro and in vivo models. The Omicron BA.1 variant showed reduced neurotropism and neurovirulence compared to Delta and D614G in human induced pluripotent stem cell (hiPSC)-derived cortical neurons co-cultured with astrocytes. Similar differences were obtained in Syrian hamsters inoculated with D614G, Delta and the Omicron BA.1 variant 5 days post infection. Replication in the olfactory mucosa was observed in all hamsters, but most prominently in D614G inoculated hamsters. Furthermore, neuroinvasion into the CNS via the olfactory nerve was observed in D614G, but not Delta or Omicron BA.1 inoculated hamsters. Furthermore, neuroinvasion was associated with neuroinflammation in the olfactory bulb of hamsters inoculated with D614G. Altogether, our findings suggest differences in the neuroinvasive, neurotropic and neurovirulent potential between SARS-CoV-2 variants using in vitro hiPSC-derived neural cultures and in vivo in hamsters during the acute phase of the infection.

Summary: In the last three decades, global production of oil palm has boomed, which has partly come at the expense of tropical rainforests. Recognizing this, many companies operating in the palm oil industry have committed to eliminate deforestation from their operations, often referred to as zero-deforestation commitments (ZDCs). Here, we estimate that if ZDCs are fully adopted and enforced across all sectors and geographies, the global extent of oil palm plantations may be 11 M ha or 40% smaller in 2030 than in a business-as-usual (BAU) scenario that assumes no compliance with ZDCs. As a result of such land-sparing effects, we estimate that 96 M ha of forests are saved from conversion, of which, 17% would otherwise have been converted (directly or indirectly) due to expanding oil palm plantations. Overall, these figures suggest that ZDCs have the potential to deliver major environmental benefits if they are fully adopted and enforced.

Summary: Background: Violence against children affects over one billion children globally. International organisations promote parenting interventions as a main strategy to reduce violence against children. Parenting interventions have therefore been implemented rapidly across the globe. Yet, evidence for their longer-term effects remains unclear. We integrated global evidence to estimate effects over time of parenting interventions to reduce physical and emotional violence against children. Methods: In this systematic review and meta-analysis, we searched 26 databases and trial registries (14 non-English: Spanish, Chinese, Farsi, Russian, Thai) and conducted an extensive grey literature search up to August 01, 2022. We included randomised controlled trials (RCTs) of parenting interventions based on social learning theory for parents of children aged 2–10 years, without time or context restrictions. We critically appraised studies using Cochrane's Risk of Bias Tool. Data were synthesised using robust variance estimation meta-analyses. This study is registered with PROSPERO, CRD42019141844. Findings: We screened 44,411 records and included 346 RCTs. Sixty RCTs reported outcomes on physical or emotional violence. Trials were distributed across 22 countries (22% LMICs). Risk of bias was high for various domains. Outcome data ranged from 0 weeks to 2 years after the intervention, and was largely based on parent self-report. Parenting interventions reduced physical and emotional violent parenting behaviours immediately after the intervention (n = 42, k = 59; d = −0.46; 95% CI: −0.59, −0.33), at 1–6 months follow-up (n = 18, k = 31; d = −0.24; 95% CI: −0.37, −0.11) and at 7–24 months follow-up (n = 12, k = 19; d = −0.18; 95% CI: −0.34, −0.02), but effects were smaller over time. Interpretation: Our findings suggest that parenting interventions can reduce physical and emotional violence against children. Effects are maintained up to 24 months follow-up, but with diminished effect sizes. With global policy interest and imminent importance, research beyond 2 years and how effects can be better sustained over time is urgently needed. Funding: Student scholarship from the Economic Social Research Council, Clarendon, and Wolfson Isaiah Berlin Fund.

Closure of ocular wounds after an accident or surgery is typically performed by suturing, which is associated with numerous potential complications, including suture breakage, inflammation, secondary neovascularization, erosion to the surface and secondary infection, and astigmatism; for example, more than half of post-corneal transplant infections are due to suture related complications. Tissue adhesives provide promising substitutes for sutures in ophthalmic surgery. Ocular adhesives are not only intended to address the shortcomings of sutures, but also designed to be easy to use, and can potentially minimize post-operative complications. Herein, recent progress in the design, synthesis, and application of ocular adhesives, along with their advantages, limitations, and potential are discussed. This review covers two main classes of ocular adhesives: (1) synthetic adhesives based on cyanoacrylates, polyethylene glycol (PEG), and other synthetic polymers, and (2) adhesives based on naturally derived polymers, such as proteins and polysaccharides. In addition, different technologies to cover and protect ocular wounds such as contact bandage lenses, contact lenses coupled with novel technologies, and decellularized corneas are discussed. Continued advances in this area can help improve both patient satisfaction and clinical outcomes.

Objectives Research suggests that military personnel frequently delay disclosing mental health issues and illness (MHI), including substance use disorder, to supervisors. This delay causes missed opportunities for support and workplace accommodations which may help to avoid adverse occupational outcomes. The current study aims to examine disclosure-related beliefs, attitudes and needs, to create a better understanding of personnel’s disclosure decision making.Design A cross-sectional questionnaire study among military personnel with and without MHI. Beliefs, attitudes and needs regarding the (non-)disclosure decision to a supervisor were examined, including factors associated with (non-)disclosure intentions and decisions. Descriptive and regression (logistic and ordinal) analyses were performed.Setting The study took place within the Dutch military.Participants Military personnel with MHI (n=324) and without MHI (n=554) were participated in this study.Outcome measure (Non-)disclosure intentions and decisions.Results Common beliefs and attitudes pro non-disclosure were the preference to solve one’s own problems (68.3%), the preference for privacy (58.9%) and a variety of stigma-related concerns. Common beliefs and attitudes pro disclosure were that personnel wanted to be their true authentic selves (93.3%) and the desire to act responsibly towards work colleagues (84.5%). The most reported need for future disclosure (96.8%) was having a supervisor who shows an understanding for MHI. The following factors were associated both with non-disclosure intentions and decisions: higher preference for privacy (OR (95% CI))=(1.99 (1.50 to 2.65)intention, 2.05 (1.12 to 3.76)decision) and self-management (OR (95% CI))=(1.64 (1.20 to 2.23)intention, 1.79 (1.00 to 3.20)decision), higher stigma-related concerns (OR (95% CI))=(1.76 (1.12 to 2.77)intention, 2.21 (1.02 to 4.79)decision) and lower quality of supervisor–employee relationship (OR (95% CI))=(0.25 (0.15 to 0.42)intention, 0.47 (0.25 to 0.87)decision).Conclusion To facilitate (early-)disclosure to a supervisor, creating opportunities for workplace support, interventions should focus on decreasing stigma and discrimination and align with personnels’ preference for self-management. Furthermore, training is needed for supervisors on how to recognise, and effectively communicate with, personnel with MHI. Focus should also be on improving supervisor–employee relationships.

Abstract Engineered living microtissues such as cellular spheroids and organoids have enormous potential for the study and regeneration of tissues and organs. Microtissues are typically engineered via self‐assembly of adherent cells into cellular spheroids, which are characterized by little to no cell–material interactions. Consequently, 3D microtissue models currently lack structural biomechanical and biochemical control over their internal microenvironment resulting in suboptimal functional performance such as limited stem cell differentiation potential. Here, this work report on stimuli‐responsive cell‐adhesive micromaterials (SCMs) that can self‐assemble with cells into 3D living composite microtissues through integrin binding, even under serum‐free conditions. It is demonstrated that SCMs homogeneously distribute within engineered microtissues and act as biomechanically and biochemically tunable designer materials that can alter the composite tissue microenvironment on demand. Specifically, cell behavior is controlled based on the size, stiffness, number ratio, and biofunctionalization of SCMs in a temporal manner via orthogonal secondary crosslinking strategies. Photo‐based mechanical tuning of SCMs reveals early onset stiffness‐controlled lineage commitment of differentiating stem cell spheroids. In contrast to conventional encapsulation of stem cell spheroids within bulk hydrogel, incorporating cell‐sized SCMs within stem cell spheroids uniquely provides biomechanical cues throughout the composite microtissues’ volume, which is demonstrated to be essential for osteogenic differentiation.

Given the dynamic nature of engineered vascular networks within biofabricated tissue analogues, it is instrumental to have control over the constantly evolving biochemical cues within synthetic matrices throughout tissue remodeling. Incorporation of pro-angiogenic vascular endothelial growth factor (VEGF165) specific aptamers into cell-instructive polymer networks is shown to be pivotal for spatiotemporally controlling the local bioactivity of VEGF that selectively elicit specific cell responses. To harness this effect and quantitatively unravel its spatial resolution, herein, bicomponent micropatterns consisting of VEGF165 specific aptamer-functionalized gelatin methacryloyl (GelMA) (aptamer regions) overlaid with pristine GelMA regions using visible-light photoinitiators (Ru/SPS) were fabricated via two-step photopatterning approach. For the 3D co-culture study, human umbilical vein-derived endothelial cells and mesenchymal stromal cells were used as model cell types. Bicomponent micropatterns with spatially defined spacings (300/500/800 ​μm) displayed high aptamer retention, aptamer-fluorescent complementary sequence (CSF) molecular recognition and VEGF sequestration localized within patterned aptamer regions. Stiffness gradient at the interface of aptamer and GelMA regions was observed with high modulus in aptamer region followed by low stiffness GelMA regions. Leveraging aptamer-tethered VEGF's dynamic affinity interactions with CS that upon hybridization facilitates triggered VEGF release, co-culture studies revealed unique characteristics of aptamer-tethered VEGF to form spatially defined luminal vascular networks covered with filopodia-like structures in vitro (spatial control) and highlights their ability to control network properties including orientation over time using CS as an external trigger (temporal control). Moreover, the comparison of single and double exposed regions within micropatterns revealed differences in cell behavior among both regions. Specifically, the localized aptamer-tethered VEGF within single exposed aptamer regions exhibited higher cellular alignment within the micropatterns till d5 of culture. Taken together, this study highlights the potential of photopatterned aptamer-tethered VEGF to spatiotemporally regulate vascular morphogenesis as a tool for controlling vascular remodeling in situ.

Central nervous system (CNS) disease is the most common extra-respiratory tract complication of influenza A virus infections in humans. Remarkably, zoonotic highly pathogenic avian influenza (HPAI) H5N1 virus infections are more often associated with CNS disease than infections with seasonal influenza viruses. Evolution of avian influenza viruses has been extensively studied in the context of respiratory infections, but evolutionary processes in CNS infections remain poorly understood. We have previously observed that the ability of HPAI A/Indonesia/5/2005 (H5N1) virus to replicate in and spread throughout the CNS varies widely between individual ferrets. Based on these observations, we sought to understand the impact of entrance into and replication within the CNS on the evolutionary dynamics of virus populations. First, we identified and characterized three substitutions-PB1 E177G and A652T and NP I119M - detected in the CNS of a ferret infected with influenza A/Indonesia/5/2005 (H5N1) virus that developed a severe meningo-encephalitis. We found that some of these substitutions, individually or collectively, resulted in increased polymerase activity in vitro. Nevertheless, in vivo, the virus bearing the CNS-associated mutations retained its capacity to infect the CNS but showed reduced dispersion to other anatomical sites. Analyses of viral diversity in the nasal turbinate and olfactory bulb revealed the lack of a genetic bottleneck acting on virus populations accessing the CNS via this route. Furthermore, virus populations bearing the CNS-associated mutations showed signs of positive selection in the brainstem. These features of dispersion to the CNS are consistent with the action of selective processes, underlining the potential for H5N1 viruses to adapt to the CNS.

Abstract Embedded 3D bioprinting has great value for the freeform fabrication of living matter. However, embedded 3D bioprinting is currently limited to highly viscous liquid baths or liquid‐like solid baths. In contrast, prior to crosslinking, most hydrogels are formulated as low‐viscosity solutions and are therefore not directly compatible with bioprinting due to low shape fidelity and poor print stability. The authors here present a method to enable low‐viscosity ink 3D (LoV3D) bioprinting, based on aqueous two‐phase stabilization of the ink‐bath interface. LoV3D allows for the printing of living constructs at high extrusion speeds (up to 1.8 m s−1) with high viability due to its exceedingly low‐viscosity. Moreover, LoV3D liquid/liquid interfaces offer unique advantages for fusing printed structures, creating intricate vasculature, and modifying surfaces at higher efficiencies than traditional systems. Furthermore, the low interfacial tension of LoV3D bioprinting offers unprecedented nozzle‐independent control over filament diameter via large‐dimension strand‐thinning, which allows for the printing of an exceptionally wide range of diameters down to the width of a single cell. Overall, LoV3D bioprinting is a unique all‐aqueous approach with broad material compatibility without the need for rheological ink adaption, which opens new avenues of application in cell patterning, drug screening, engineered meat, and organ fabrication.

The increased demand for personalized wearable and implantable medical devices has created the need for the generation of electronics that interface with living systems. Current bioelectronics has not fully resolved mismatches between biological systems and engineered circuits, resulting in tissue injury and pain. Thus, there is an unmet need to develop materials for the fabrication of wearable electronics that are biocompatible at the tissue interface. Here, we developed a tailorable gelatin-based bio-ink functionalized with a choline bio-ionic liquid (BIL) for in situ 3D bioprinting of bioelectronics at the tissue interface. The resultant photocrosslinked polymer is programmable, transparent, ion conductive, and flexible. BILs are stably conjugated with a gelatin methacryloyl (GelMA) hydrogel using photocrosslinking to make BioGel, which routes ionic current with high resolution and enables localized electrical stimulation delivery. Controllable crosslinking, achieved by varying reactants composition, allows the BioGel bio-ink platform for easy and rapid in-situ 3D bioprinting of complex designs directly on skin tissue. Bio-ionic modified polymers thus represent a versatile and wide-applicable bio-ink solution for personalized bioelectronics fabrication that minimizes tissue damage.

Objective: We tested if baseline disruptive child behavior problem severity predicts parental attendance at sessions of a parenting group program. Method: We used a database of randomized trials of the Incredible Years parenting program in Europe and restricted the sample to participants randomized to the intervention arm. Using baseline Eyberg Child Behavior Inventory scores, we distinguished between trial-level problem severity and child-level problem severity, compared linear and quadratic functional forms at both levels, and considered cross-level interactions, all in a multilevel Poisson regression framework. Results: Drawing on 918 participants in 12 trials, we found that within trials, parents of children with the least and most severe problems attended fewer sessions. Between trials, each additional 10-point increase in the Eyberg Child Behavior Inventory trial mean predicted an 11% increase in attendance. Models including child sex, age, or family low-income did not change coefficients or their interpretation. Conclusions: Our findings suggest that although generally attendance is higher in parents of children with more challenging behavior, it seems difficult for group programs to keep families with the least or most severe problems engaged. Our findings call for the need to better understand the conditions under which lower attendance translates into equivalent or lesser program benefits. What is the public health significance of this article?: This analysis suggests that in the Incredible Years parenting program, families with the least and most severe child behavior problems attend the fewest sessions. A greater focus on keeping these families' engagement may result in larger program benefits. [ABSTRACT FROM AUTHOR]