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Abstract Background Caudal-type homeobox 2 (CDX2) and special AT-rich sequence-binding protein 2 (SATB2) are transcription factors playing important roles in intestinal homeostasis and participating in the regulation of intestinal inflammation. In colorectal cancer (CRC), reduced expression levels of CDX2 and SATB2 have been associated with poor differentiation and worse survival. However, their prognostic significance still needs further clarification, and the associations between CDX2 and SATB2 and immune cell infiltration into the CRC microenvironment are largely unknown. Methods We analyzed CDX2 and SATB2 expression in two large cohorts of stages I–IV CRC patients (N = 2302) and analyzed their associations with clinicopathologic parameters, the density of local immune cells (determined with three multiplex immunohistochemistry panels and conventional immunohistochemistry), and survival. Results In mismatch repair-proficient tumors, reduced CDX2 and SATB2 expression were associated with higher densities of immature monocytic cells, macrophages, and M2-like macrophages. Low expression of CDX2 was associated with shorter cancer-specific survival independent of conventional prognostic parameters in both cohorts. In the larger cohort, adjusted hazard ratio (HR) for negative (vs. high) CDX2 expression was 3.62 (95% CI 2.08–6.31, p trend

Abstract Background Headache conditions have a high prevalence worldwide. Office workers with high and demanding workload, but low physical activity levels are considered vulnerable for suffering from headache. This analysis examines whether exercise combined with health promotion at the workplace is effective for headache relief in office workers. Methods This study reports the results of secondary outcomes of a stepped wedge cluster randomized controlled trial. Office workers (n = 120) were randomly assigned to a twelve-week supervised intervention period, consisting of neck and shoulder girdle exercises with health promotion interventions performed at the workplace. Secondary outcomes were analysed and modelled for headache occurrence, frequency, and the Headache Impact Test-6 (HIT-6), accounting for possible effects for the intervention, the period it had been introduced, and interactional and nested effects. Results At baseline, 88 of the 120 participants reported ≥ one headache episode in the past four weeks, with a mean headache frequency of 3.58 days for that period. The mean HIT-6 score for the entire cohort amounted to 53.6 points. For headache occurrence and HIT-6, the simplest model with the intervention only, showed the best statistical fit with an odds ratio for headache occurrence of 0.46 (95% confidence interval: 0.25 to 0.84), and − 2.23 (95% confidence interval: -3.35 to -1.12) points on the HIT-6 questionnaire. For headache frequency, the model accounting for interaction effects (intervention x period) had the best statistical fit and showed an incidence rate ratio of 0.57 (95% confidence interval: 0.44 to 0.74) for the first period, but not for later ones. Conclusions Neck exercises and health promotion had a positive impact on headache occurrence, headache frequency and HIT-6, with the latter not reaching clinical importance. Although only statistically significant for headache frequency, larger effects were found during earlier periods or shorter interventional exposure for all outcomes, necessitating refresher sessions at later periods. Trial registration NCT04169646.

Abstract Tissue inhibitor of matrix metalloproteinase 1 (TIMP1) is a multifaceted, cytokine-like bioactive molecule whose levels are elevated in a wide range of inflammatory diseases and are associated with prognosis. Additionally, TIMP1 may play a role in driving systemic inflammation. TIMP1 immunohistochemistry and TIMP1 serum concentrations were analyzed in a cohort of 776 colorectal cancer patients. TIMP1 histoscore by cell type (tumor cell, other) was quantified using digital image analysis. Serum TIMP1 levels were evaluated for correlations with tumor TIMP1 expression, and their associations with tumor characteristics, inflammation, and prognosis were investigated. High serum TIMP1 concentrations associated with shorter overall survival (multivariable HR 1.85, 95% CI 1.30–2.65). Serum TIMP1 levels positively correlated with markers of systemic inflammation and tumor necrosis percentage but not with TIMP1 expression in tumor tissue. High TIMP1 intensity in tumor stroma associated with longer cancer-specific and overall survival in univariable analysis but not in multivariable models. T cell densities in tumor tissue positively correlated with tumor stromal TIMP1 expression and negatively with tumor epithelial TIMP1 expression. Serum TIMP1 levels show promise as a prognostic marker for colorectal cancer and correlate with systemic inflammatory markers, but do not correlate with TIMP1 expression in tumor tissue.

Abstract Background Fall prevention programmes are essential interventions in societies with aging populations. This study assessed the fall rate and other health outcomes, as well as the cost-effectiveness of a home-based fall prevention programme for community-dwelling older people. In a single home visit, trained physical or occupational therapists performed fall risk assessments, eliminated environmental risk factors, and provided tailored exercises. Methods A prospective, longitudinal observational study was performed with participants of a fall prevention programme who agreed to be followed-up over one year. Baseline data included self-reported falls one month and one year before the intervention. Participants were monitored through bi-monthly telephone calls, assessing their number of falls, fear of falling (using the Falls Efficacy Scale-International (FES-I), quality of life (using the EuroQuol-5 Dimensions-5 Levels, EQ-5D-5L), and physical activity (in minutes per week). Statistical analysis of the data used a Generalized Estimating Equations (GEE) Poisson-Modell for number of falls and a Linear Mixed Model (LMM) for fear of falling, quality of life and physical activity. In addition, health insurance claims data were used to compare the number of medically treated falls in the year before and after the intervention, as well as the related healthcare spending. Cost-effectiveness of the programme versus usual care was estimated as cost per prevented medically treated fall. Results Overall, 639 person-years of observation time were analyzed. Participants had a mean age of 81.8 years (+/- 5.2) and 59% were female. On average, the fall rate decreased from 1.35 to 1.02 per person-year, or -23.9% (95%CI from -35.92 to -9.67), fear of falling decreased by -1.27 points (95%CI from −1.50 to -1.05), quality of life improved by -0.88 points (95%CI from −1.09 to -0.68), and physical activity increased by 9.87 min per day (95%CI from 5.65 to 14.09). Analysis of claims data showed a 48.0% reduction (95%CI from 30.5% to 61.0%) in medically treated falls. The average cost per prevented medically treated fall was estimated at approximately 1,353 USD, with a 50% probability of the intervention being cost saving. Conclusions This fall prevention programme with a single home visit was found to be effective and cost effective. Health policies should establish such a model as a reimbursed standard care to assist in combatting the increasing burden of falls on individuals and societies.

(1) Background: Clinical aspects like sex, age, Karnofsky Performance Scale (KPS) and psychosocial distress can affect the health-related quality of life (HR-QoL) and treatment satisfaction of patients with malignant isocitrate dehydrogenase wild-type (IDHwt) gliomas and caregivers. (2) Methods: We prospectively investigated the HR-QoL and patient/caregiver treatment satisfaction in a cross-sectional study with univariable and multiple regression analyses. Questionnaires were applied to investigate the HR-QoL (EORTC QLQ-C30, QLQ-BN20) and treatment satisfaction (EORTC PATSAT-C33). (3) Results: A cohort of 61 patients was investigated. A higher KPS was significantly associated with a better HR-QoL regarding the functional scales of the EORTC QLQ-C30 (p < 0.004) and a lower symptom burden regarding the EORTC QLQ-BN20 (p < 0.001). The patient treatment satisfaction was significantly poorer in the patients older than 60 years in the domain of family involvement (p = 0.010). None of the investigated aspects showed a significant impact on the treatment satisfaction of caregivers. (4) Conclusions: We demonstrated that in patients with IDHwt gliomas, the KPS was the most important predictor for a better HR-QoL in functional domains. Data on the HR-QoL and treatment satisfaction in patients with IDHwt gliomas and their caregivers are rare; therefore, further efforts should be made to improve supportive care in this highly distressed cohort.

Summary: The study of somatic mutations in single cells provides insights into aging and carcinogenesis, which is complicated by the dependency on whole-genome amplification (WGA). Here, we describe a detailed workflow starting from single-cell isolation to WGA by primary template-directed amplification (PTA), sequencing, quality control, and downstream analyses. A machine learning approach, the PTA Analysis Toolkit (PTATO), is used to filter the hundreds to thousands of artificial variants induced by WGA from true mutations at high sensitivity and accuracy.For complete details on the use and execution of this protocol, please refer to Middelkamp et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

IntroductionPsychosocial distress and mental health problems are common in patients before and after solid organ transplantation and can negatively affect morbidity, mortality, and adherence. Even though regular screening is recommended to identify patients with high levels of distress, the implementation in routine care has been insufficient so far.MethodsTwo newly developed problem lists for patients before and after transplantations were pilot implemented for 8 weeks at the Medical Center Hamburg Eppendorf (UKE) to identify factors facilitating and impeding their implementation.ResultsHealth care professionals evaluated its appropriateness, feasibility, and the cooperation with the psychologists before (HCPs: n = 23) and after (HCPs: n = 19) the implementation. Four psychologists assessed the appropriateness and feasibility by answering to open-ended and close-ended questions. Additionally, patients before (n = 8) and after (n = 100) transplantation filled out the screening and rated its acceptance. Only the data of the patients after transplantation were analyzed due to the small sample size of patients before transplantation. HCPs and psychologists rated the screenings as very appropriate [HCPs: M = 3.84 (SD = 0.77) to M = 4.32 (SD = 0.58)]. It was also highly accepted among patients [M = 4.23 (SD = 0.85) to M = 4.68 (SD = 0.65)]. Contentment with the psychological support and understanding of the mental health problems among HCPs increased significantly from before to after the implementation (U = 107.50, p < 0.05, r = 0.33; U = 107.00, p < 0.05, r = 0.34). The feasibility of the problem list post-Tx in routine care, however, was seen as challenging [HCPs: M = 3.11 (SD = 1.05) to M = 3.47 (SD = 1.07)].DiscussionThe distress screening was accepted and improved the cooperation between different professions. Barriers to implementation can be lack of staff and resources. Future studies should assess the adoption and sustainability of the screening in routine care.

Abstract Background Microglial activation is one hallmark of Alzheimer disease (AD) neuropathology but the impact of the regional interplay of microglia cells in the brain is poorly understood. We hypothesized that microglial activation is regionally synchronized in the healthy brain but experiences regional desynchronization with ongoing neurodegenerative disease. We addressed the existence of a microglia connectome and investigated microglial desynchronization as an AD biomarker. Methods To validate the concept, we performed microglia depletion in mice to test whether interregional correlation coefficients (ICCs) of 18 kDa translocator protein (TSPO)-PET change when microglia are cleared. Next, we evaluated the influence of dysfunctional microglia and AD pathophysiology on TSPO-PET ICCs in the mouse brain, followed by translation to a human AD-continuum dataset. We correlated a personalized microglia desynchronization index with cognitive performance. Finally, we performed single-cell radiotracing (scRadiotracing) in mice to ensure the microglial source of the measured desynchronization. Results Microglia-depleted mice showed a strong ICC reduction in all brain compartments, indicating microglia-specific desynchronization. AD mouse models demonstrated significant reductions of microglial synchronicity, associated with increasing variability of cellular radiotracer uptake in pathologically altered brain regions. Humans within the AD-continuum indicated a stage-depended reduction of microglia synchronicity associated with cognitive decline. scRadiotracing in mice showed that the increased TSPO signal was attributed to microglia. Conclusion Using TSPO-PET imaging of mice with depleted microglia and scRadiotracing in an amyloid model, we provide first evidence that a microglia connectome can be assessed in the mouse brain. Microglia synchronicity is closely associated with cognitive decline in AD and could serve as an independent personalized biomarker for disease progression.

Abstract Background Osteoporotic fractures signal severely compromised bone strength and are associated with a greatly increased risk of refracture. Despite the availability of effective and safe medications that reduce fracture risk, 70–80% of patients are inadequately investigated or treated for osteoporosis following an initial fracture, constituting a significant ‘osteoporosis care gap’. Optimal methods of bridging this gap with primary care at the forefront of secondary fracture prevention remain undetermined. This protocol describes a cluster randomised controlled trial to evaluate the effectiveness of a novel integrated model of secondary fracture prevention and management in primary care. Methods The cluster randomised controlled trial involves multiple branches of a community-based radiology provider (CRP), a hospital-based secondary fracture prevention program (SFPP) and numerous primary care practices in metropolitan Sydney that refer to either the CRP or SFPP. Using natural language processing tools, patients diagnosed with a potential osteoporotic fracture will be identified by automatically screening radiology reports generated at the CRP or SFPP. The primary care practices that these patients attend will be randomised (1:1) to either the intervention or usual care. The intervention consists of (i) electronic and fax alerts informing the practice/primary care physician that their patient has been diagnosed with a potential osteoporotic fracture; (ii) provision of osteoporosis management guidelines and (iii) follow-up surveys at 4 weeks and 6 months. Practices in the usual care (control) group will receive no alerts and provide usual care. The primary outcome is the proportion of patients undergoing a bone density scan and/or filling a prescription for osteo-protective pharmacotherapy within 3 months of the initial diagnostic imaging report. Secondary outcomes are the proportion of patients: (i) undergoing an osteoporosis-related blood test within 3 months of the initial diagnostic imaging report; (ii) initiated on a chronic disease management plan within 3 months of the diagnostic report, and (iii) filling a second prescription for osteo-protective pharmacotherapy within 9 months post initial diagnostic imaging report. Outcomes will be obtained through de-identified linked data from Medical Benefits Schedule and Pharmaceutical Benefits Scheme held by the Australian Institute of Health and Welfare. Discussion This is the first randomised trial to integrate case-detection of potential osteoporotic fractures in a hospital and community setting with direct alerts to the patient’s primary care provider. This study will determine whether such an intervention is effective in improving investigation and/or treatment rates of osteoporosis in patients with a potential osteoporotic fracture. Trial registration This study is registered with the Australian New Zealand Clinical Trials Registry (ANZCTR): ACTRN12623000658617p.

Abstract Background Several recent global events may have impacted adolescent sleep and exacerbated pre-existing disparities by social positions (i.e., social roles, identity or sociodemographic factors, and/or group memberships that are associated with power and oppression due to the structures and processes in a given society at given time). Current understanding of sleep among adolescents is critical to inform interventions for a more equitable future, given the short and long-term consequences of inadequate sleep on health and well-being. This study aimed to provide contemporary evidence on sleep disparities by key social positions among adolescents in Canada. Methods Cross-sectional analyses were conducted using self-reported data collected during 2020–2021 (the first full school year after the COVID-19 pandemic onset) from 52,138 students (mean [SD] age = 14.9 [1.5]) attending 133 Canadian secondary schools. Multiple regression models were used to test whether sleep quality (how well students slept during past week), duration (weekday, weekend, weighted daily average), and guideline adherence (8–10 h/day) differed by sex and gender, race and ethnicity, and socioeconomic status (SES). Results Females reported a mean [95% CI] difference of -1.7 [-3.7, 0.4] min/day less sleep on weekdays than males, but 7.1 [4.5, 9.6] min/day more sleep on weekends, resulting in no difference in average daily sleep between males and females. Females were less likely to report good quality sleep compared to males (AOR = 0.57 [0.54, 0.60]). SES followed a generally monotonic trend where higher scores were associated with more sleep on weekdays (Δhighest: lowest = -28.6 [-39.5, -17.6]) and weekends (Δhighest: lowest = -17.5 [-3.8, -31.2]) and greater likelihood of higher sleep quality (AORhighest: lowest = 3.04 [2.35, 3.92]). Relative to White adolescents, weekday and average daily sleep duration were lower among all other racial identities; mean differences ranged from ∼ 5–15 min/day, with Black students reporting the least sleep. Conclusions Differences in sleep duration and quality were most profound among adolescents from the lowest and highest SES. Racial disparities were more evident on weekdays. Compensatory weekend sleep appears more pronounced in females than males. Addressing sleep inequities is critical, as a robust predictor of multiple health outcomes.

Abstract Therapy-related myeloid neoplasms (t-MN) arise as a complication of chemo- and/or radiotherapy. Although t-MN can occur both in adult and childhood cancer survivors, the mechanisms driving therapy-related leukemogenesis likely vary across different ages. Chemotherapy is thought to induce driver mutations in children, whereas in adults pre-existing mutant clones are selected by the exposure. However, selective pressures induced by chemotherapy early in life are less well studied. Here, we use single-cell whole genome sequencing and phylogenetic inference to show that the founding cell of t-MN in children starts expanding after cessation of platinum exposure. In patients with Li-Fraumeni syndrome, characterized by a germline TP53 mutation, we find that the t-MN already expands during treatment, suggesting that platinum-induced growth inhibition is TP53-dependent. Our results demonstrate that germline aberrations can interact with treatment exposures in inducing t-MN, which is important for the development of more targeted, patient-specific treatment regimens and follow-up.

We describe the R package BEKKs, which implements the estimation and diagnostic analysis of a prominent family of multivariate generalized autoregressive conditionally heteroskedastic (MGARCH) processes, the so-called BEKK models. Unlike existing software packages, we make use of analytical derivatives implemented in efficient C++ code for nonlinear log-likelihood optimization. This allows fast parameter estimation even in higher model dimensions N > 3. The baseline BEKK model is complemented with an asymmetric parameterization that allows for a flexible modeling of conditional (co)variances. Furthermore, we provide the user with the simplified scalar and diagonal BEKK models to deal with high dimensionality of heteroskedastic time series. The package is designed in an object-oriented way featuring a comprehensive toolbox of methods to investigate and interpret, for instance, volatility impulse response functions, risk estimation and forecasting (VaR) and a backtesting algorithm to compare the forecasting performance of alternative BEKK models. For illustrative purposes, we analyze a bivariate ETF return series (S&P, US treasury bonds) and a four-dimensional system comprising, in addition, a gold ETF and changes of a log oil price by means of the suggested package. We find that the BEKKs package is more than 100 times faster for time series systems of dimension N > 3 than other existing packages.

PurposeThe development of brain metastases (BM) can significantly increase the psycho-oncological burden in cancer patients, requiring timely intervention. In addition, this aspect may negatively affect the course of the disease and treatment outcome. However, screening for psycho-oncological burden is often overlooked in clinical routine. Therefore, we analyzed the extent of psycho-oncological distress in a patient population with BM receiving neurosurgical resection and identified clinical characteristics associated with a high need for psycho-oncological intervention.MethodsWe prospectively screened 353 patients (169 female, 184 male, mean age 61.9 years) scheduled for microsurgical resection of one or more BM. Psycho-oncological screening was performed on the day of admission using the Hornheider screening instrument (HSI) and the distress thermometer (DT). Screening results were correlated with demographic and clinical data.ResultsMost patients (73.1%) completed the screening questionnaire. Patients who failed to complete the questionnaire presented more frequently with metachronous BM (74.7% vs. 25.3%, p=0.009), were significantly older (p=0.0018), and had a significantly lower KPS score (p=0.0002). Based on the threshold values of the questionnaires, 59.3% of the patients showed a significant psycho-oncological burden requiring immediate intervention. Univariate analysis demonstrated that synchronous BM (p=0.034), tumors in eloquent areas (p=0.001), lower KPS (p=0.031), female gender (p=0.009), and presurgical aphasia (p=0.042) were significantly associated with high psycho-oncological burden. Multivariate analysis showed synchronous BM (p=0.045), female gender (p=0.005), and lower KPS (p=0.028) as independent factors associated with high psycho-oncological burden.ConclusionThe majority of patients with BM have a high psycho-oncological burden. Female gender, synchronous BM, and lower KPS are independently associated with a need for psycho-oncological intervention.

IntroductionThe oligosaccharide 2′-fucosyllactose (2′-FL) is a predominant component of human milk, serving as a prebiotic for gut microbiota and influencing immune development in infants. Bifidobacterium longum subspecies infantis (B. infantis) is a commensal bacterium found in breastfed infants. Both 2′-FL and a specific strain of B. infantis, Bi-26™, are commercially available. This study investigates the potential synbiotic relationship between 2′-FL and Bi-26™ on immune development.MethodsTwo-day-old piglets (n = 53) were randomized in a 2 × 2 design, receiving either a commercial milk replacer ad libitum without (CON) or with 1.0 g/L 2′-FL (FL). Piglets in each diet were further randomized to receive either glycerol stock alone or Bi-26™ (109 CFU) (BI and FLBI) orally once daily. On postnatal day (PND) 34/35, animals were euthanized, and blood was collected for serum cytokine analysis. Additionally, peripheral blood mononuclear cells (PBMCs) were isolated for ex vivo stimulation and flow cytometry analysis. Serum and ex vivo cytokines were analyzed using a multivariate model. All other outcomes were analyzed using a two-way ANOVA, considering prebiotic and probiotic fixed effects. The significance level was set at a p value

Abstract Cardiac magnetic resonance (CMR) imaging allows precise non-invasive quantification of cardiac function. It requires reliable image segmentation for myocardial tissue. Clinically used software usually offers automatic approaches for this step. These are, however, designed for segmentation of human images obtained at clinical field strengths. They reach their limits when applied to preclinical data and ultrahigh field strength (such as CMR of pigs at 7 T). In our study, eleven animals (seven with myocardial infarction) underwent four CMR scans each. Short-axis cine stacks were acquired and used for functional cardiac analysis. End-systolic and end-diastolic images were labelled manually by two observers and inter- and intra-observer variability were assessed. Aiming to make the functional analysis faster and more reproducible, an established deep learning (DL) model for myocardial segmentation in humans was re-trained using our preclinical 7 T data (n = 772 images and labels). We then tested the model on n = 288 images. Excellent agreement in parameters of cardiac function was found between manual and DL segmentation: For ejection fraction (EF) we achieved a Pearson’s r of 0.95, an Intraclass correlation coefficient (ICC) of 0.97, and a Coefficient of variability (CoV) of 6.6%. Dice scores were 0.88 for the left ventricle and 0.84 for the myocardium.

Abstract Robust chronologies and time equivalent tephra markers are essential to better understand spatial palaeoenvironmental response to past abrupt climatic changes. Identification of well-dated and widely dispersed volcanic ash by tephra and cryptotephra (microscopic volcanic ash) provides time synchronous tie-points and strongly reduces chronological uncertainties. Here, we present the major, minor and trace element analyses of cryptotephra shards in the Dead Sea Deep Drilling sedimentary record (DSDDP 5017-1A) matching the Campanian Ignimbrite (CI). This geochemical identification expands the known dispersal range of the CI to the southeastern Mediterranean, over 2300 km from the volcanic source. Due to the CI eruption occurring near-synchronous with North Atlantic ice surge of Heinrich Event 4 (HE4), this tephra provides insights into regional responses to large-scale climatic change in the Mediterranean. In the Dead Sea, the CI layer is associated with wetter climatic conditions. This contrasts with the contemporaneous occurrence of the CI deposition and dry conditions in the central and eastern Mediterranean suggesting a possible climate time-transgressive expansion of HE4. Our finding underscores the temporal and spatial complexity of regional climate responses and emphasises the importance of tephra as a time marker for studying large-scale climatic changes verses regional variations.

The occurrence frequency distributions of fluxes ( F ) and fluences or energies ( E ) observed in the majority (in 18 out of 23 cases) of astrophysical phenomena are found to be consistent with the predictions of the fractal-diffusive self-organized criticality (FD-SOC) model, which predicts power-law slopes with universal constants of α _F = (9/5) = 1.80 for the flux and α _E = (5/3) ≈ 1.67 for the fluence. The theoretical FD-SOC model is based on the fractal dimension, the flux-volume proportionality, and classical diffusion. The universal scaling laws predict the size distributions of numerous astrophysical phenomena, such as solar flares, stellar flares, coronal mass ejections, auroras, blazars, active galactic nuclei, black hole systems, galactic fast radio bursts, gamma-ray bursts, and soft gamma-ray repeaters. In contrast, we identify five outliers of astrophysical phenomena, i.e., coherent solar radio bursts, random solar radio bursts, solar energetic particles, cosmic rays, and pulsar glitches, which are not consistent with the standard FD-SOC model, and thus require different physical mechanisms.

Abstract Ultra-low-dose computed tomography (ULD-CT) may combine the high sensitivity of conventional computed tomography (CT) in detecting sarcoma pulmonary metastasis, with a radiation dose in the same magnitude as chest X-ray (CXR). Fifty patients with non-metastatic high-grade soft tissue sarcoma treated with curative intention were recruited. Their follow-up involved both CXR and ULD-CT to evaluate their different sensitivity. Suspected findings were confirmed by conventional CT if necessary. Patients with isolated pulmonary metastases were treated with surgery or stereotactic body radiation therapy (SBRT) with curative intent if possible. The median effective dose from a single ULD-CT study was 0.27 mSv (range 0.12 to 0.89 mSv). Nine patients were diagnosed with asymptomatic lung metastases during the follow-up. Only three of them were visible in CXR and all nine in ULD-CT. CXR had therefore only a 33% sensitivity compared to ULD-CT. Four patients were operated, and one had SBRT to all pulmonary lesions. Eight of them, however, died of the disease. Two patients developed symptomatic metastatic recurrence involving extrapulmonary sites+/−the lungs between two imaging rounds. ULD-CT has higher sensitivity for the detection of sarcoma pulmonary metastasis than CXR, with a radiation dose considerably lower than conventional CT. Clinical trial registration: NCT05813808. 04-14-2023.

Abstract Reactive Oxygen Species (ROS) derived from mitochondrial respiration are frequently cited as a major source of chromosomal DNA mutations that contribute to cancer development and aging. However, experimental evidence showing that ROS released by mitochondria can directly damage nuclear DNA is largely lacking. In this study, we investigated the effects of H2O2 released by mitochondria or produced at the nucleosomes using a titratable chemogenetic approach. This enabled us to precisely investigate to what extent DNA damage occurs downstream of near- and supraphysiological amounts of localized H2O2. Nuclear H2O2 gives rise to DNA damage and mutations and a subsequent p53 dependent cell cycle arrest. Mitochondrial H2O2 release shows none of these effects, even at levels that are orders of magnitude higher than what mitochondria normally produce. We conclude that H2O2 released from mitochondria is unlikely to directly damage nuclear genomic DNA, limiting its contribution to oncogenic transformation and aging.

IntroductionThe COVID-19 pandemic intensified the impact of risk factors for adolescent mental health, including financial worry. Social support has shown to protect from negative mental health during times of stress. We examined the effect of financial worry on changes in anxiety and depression symptoms among Canadian adolescents prior to and during the pandemic, and assessed whether social support from family and friends moderated any changes. MethodsWe analyzed 2-year linked data from the 2018/19 (pre-pandemic) and 2020/21 (during-pandemic) waves of the COMPASS study, with reports from 12 995 Canadian secondary school students. A series of multilevel linear regressions were conducted to examine the main hypotheses under study. ResultsStudents scored an average (SD) of 7.2 (5.8) on the anxiety (GAD-7) and 10.0 (6.5) on the depression (CESD-10) scales; 16.1% reported they experienced financial worry during the pandemic. Financial worry was a strong and significant predictor of increased anxiety scores (+1.7 score between those reporting “true/mostly true” versus “false/mostly false”) during the pandemic, but not for depression scores. Low family and friend support were associated with anxiety, and low family support was associated with depression. No significant interactions were detected between social support and financial worry. ConclusionPandemic-related financial worry was significantly associated with anxiety in our large sample of Canadian adolescents. Clinical and public health initiatives should be aware of adolescents’ financial worry and its associations with anxiety during times of crisis.

IntroductionLa pandémie de COVID-19 a intensifié les effets des facteurs de risque associés à la santé mentale des adolescents, en particulier les préoccupations financières. Par ailleurs, on sait que le soutien social offre une protection contre les problèmes de santé mentale pendant les périodes de stress. C’est dans ce cadre que nous avons exploré l’effet des préoccupations financières sur les variations des symptômes d’anxiété et de dépression chez les adolescents canadiens avant et pendant la pandémie, pour déterminer si le soutien social apporté par la famille et les amis modérait ces variations. MéthodologieNous avons analysé des données couplées sur deux ans provenant des vagues 2018-2019 (avant la pandémie) et 2020-2021 (pendant la pandémie) de l’étude COMPASS pour 12 995 élèves canadiens du secondaire. Nous avons réalisé une série de régressions linéaires multiniveaux pour explorer les principales hypothèses à l’étude. RésultatsLes élèves ont obtenu un score moyen de 7,2 (écart-type : 5,8) pour l’anxiété (échelle GAD-7) et de 10,0 (6,5) pour la dépression (échelle CESD-10), et 16,1 % des élèves ont déclaré avoir eu des préoccupations financières pendant la pandémie. Les préoccupations financières ont constitué un facteur de prédiction important et statistiquement significatif de scores d’anxiété plus élevés (+1,7 entre ceux qui ont répondu « vrai/le plus souvent vrai » et ceux qui ont répondu « faux/le plus souvent faux ») pendant la pandémie, mais non en ce qui concerne les scores de dépression. Il existe un lien entre un faible soutien de la part de la famille et des amis et l’anxiété ainsi qu’entre un faible soutien de la part de la famille et la dépression. Aucune interaction importante n’a été observée entre le soutien social et les préoccupations financières. ConclusionLes préoccupations financières liées à la pandémie ont été fortement associées à l’anxiété dans notre large échantillon d’adolescents canadiens. Les initiatives cliniques et sanitaires doivent tenir compte des préoccupations financières des adolescents et de leurs liens avec l’anxiété en période de crise.

Abstract Background To assess whether changes in breakfast and water consumption during the first full school year after the emergence of the COVID-19 pandemic varied based on sex/gender, race/ethnicity, and socioeconomic status among Canadian adolescents. Methods Prospective annual survey data collected pre- (October 2019-March 2020) and post-COVID-19 onset (November 2020-June 2021) the Cannabis, Obesity, Mental health, Physical activity, Alcohol, Smoking, and Sedentary behaviour (COMPASS) study. The sample consisted of 8,128 students; mean (SD) age = 14.2 (1.3) years from a convenience sample of 41 Canadian secondary schools. At both timepoints self-reported breakfast and water consumption were dichotomized as daily or not. Multivariable logistic generalized estimating equations with school clustering were used to estimate differences in maintenance/adoption of daily consumption post-COVID-19 based on demographic factors, while controlling for pre-COVID-19 behaviour. Results Adjusted odds ratios (AOR) with 95% confidence intervals are reported. Females (AOR = 0.71 [0.63, 0.79]) and lower socioeconomic status individuals (AORLowest:Highest=0.41 [0.16, 1.00]) were less likely to maintain/adopt daily breakfast consumption than male and higher socioeconomic status peers in the 2020–2021 school year. Black identifying individuals were less likely than all other racial/ethnic identities to maintain/adopt plain water consumption every day of the week (AOR = 0.33 [0.15, 0.75], p

Abstract One of the most critical steps for accurate taxonomic identification in DNA (meta)-barcoding is to have an accurate DNA reference sequence dataset for the marker of choice. Therefore, developing such a dataset has been a long-term ambition, especially in the Viridiplantae kingdom. Typically, reference datasets are constructed with sequences downloaded from general public databases, which can carry taxonomic and other relevant errors. Herein, we constructed a curated (i) global dataset, (ii) European crop dataset, and (iii) 27 datasets for the EU countries for the ITS2 barcoding marker of vascular plants. To that end, we first developed a pipeline script that entails (i) an automated curation stage comprising five filters, (ii) manual taxonomic correction for misclassified taxa, and (iii) manual addition of newly sequenced species. The pipeline allows easy updating of the curated datasets. With this approach, 13% of the sequences, corresponding to 7% of species originally imported from GenBank, were discarded. Further, 259 sequences were manually added to the curated global dataset, which now comprises 307,977 sequences of 111,382 plant species.

Previous research has examined postsecondary student mental health and transitions into university. However, research focused on the transition out of university is lacking. Challenges may be experienced differently among population subgroups. We examined the mental health and support access of university students approaching graduation and differences by various social positions. Survey data were collected from final-year undergraduate students that had registered to graduate at a Canadian university in 2021 and 2022. Chi-square and linear regression models analyzed relationships between sociodemographic characteristics and mental health outcomes. Open-ended questions assessed barriers to accessing support and desired supports. Sexual/gender diverse students reported greater depressive symptoms than cisgender heterosexual students. Students without stressful childhood or current financial situations had lower depression and anxiety scores than their peers that experienced stressful financial situations, respectively. Formal support was more commonly accessed off-campus than on-campus in the past year. Differences in past-year support access were found by gender/sexuality, financial stress, age and race/ethnicity. Availability/scheduling was the most reported barrier to accessing campus-based services. Financial concerns were a common challenge and area for desired support. Implementing developmentally specific mental health support catering to the demands of this life period is necessary.

Abstract CHESS 3 represents an improved human gene catalog based on nearly 10,000 RNA-seq experiments across 54 body sites. It significantly improves current genome annotation by integrating the latest reference data and algorithms, machine learning techniques for noise filtering, and new protein structure prediction methods. CHESS 3 contains 41,356 genes, including 19,839 protein-coding genes and 158,377 transcripts, with 14,863 protein-coding transcripts not in other catalogs. It includes all MANE transcripts and at least one transcript for most RefSeq and GENCODE genes. On the CHM13 human genome, the CHESS 3 catalog contains an additional 129 protein-coding genes. CHESS 3 is available at http://ccb.jhu.edu/chess .

Abstract Cardiovascular disease (CVD) is one of the leading causes of premature retirement. However, the relationship between CVD risk factors and workforce participation is not well known. We studied the relationship between midlife CVD risk, age at retirement, work-loss years, and survival in retirement. Middle-aged Finnish men (initial n = 3490, mean age = 47.8 years) were assessed for CVD risk factors and general health in the 1970s. They worked as business executives and provided information on their retirement status in the year 2000. Survival was followed up to the 9th decade of life with a follow-up of up to 44 years. Work-loss years were calculated as death or retirement occurring at age ≤ 65 years. Smoking, body mass index, and alcohol use were used as covariates, excluding models of CVD risk, which were adjusted for alcohol use only. Higher risk of 10-year fatal CVD was associated with 0.32 more years (relative risk

Traditionally, two fundamentally different theoretical approaches have been used in emotion research to model (human) emotions: discrete emotion theories and dimensional approaches. More recent neurophysiological models like the hierarchical emotion theory suggest that both should be integrated. The aim of this review is to provide neurocognitive evidence for this perspective with a particular focus on experimental studies manipulating anxiety and/or curiosity. We searched for evidence that the neuronal correlates of discrete and dimensional emotional systems are tightly connected. Our review suggests that the ACC (anterior cingulate cortex) responds to both, anxiety, and curiosity. While amygdala activation has been primarily observed for anxiety, at least the NAcc (nucleus accumbens) responds to both, anxiety and curiosity. When these two areas closely collaborate, as indicated by strong connectivity, this may indicate emotion regulation, particularly when the situation is not predictable.

We report a mixture of expert strategy to create fine-tuned large language models using a deep layer-wise token-level approach based on low-rank adaptation (LoRA). Starting with a set of pre-trained LoRA adapters, our gating strategy uses the hidden states to dynamically mix adapted layers, allowing the resulting X-LoRA model to draw upon different capabilities and create never-before-used deep layer-wise combinations to solve tasks. The design is inspired by the biological principles of universality and diversity, where neural network building blocks are reused in different hierarchical manifestations. Hence, the X-LoRA model can be easily implemented for any existing large language model without a need for modifications of the underlying structure. We develop a tailored X-LoRA model that offers scientific capabilities, including forward/inverse analysis tasks and enhanced reasoning capability, focused on biomaterial analysis, protein mechanics, and design. The impact of this work includes access to readily expandable and adaptable models with strong domain knowledge and the capability to integrate across areas of knowledge. Featuring experts in biology, mathematics, reasoning, bio-inspired materials, mechanics and materials, chemistry, protein biophysics, mechanics, and quantum-mechanics based molecular properties, we conduct a series of physics-focused case studies. We examine knowledge recall, protein mechanics forward/inverse tasks, protein design, adversarial agentic modeling including ontological knowledge graph construction, and molecular design. The model is capable not only of making quantitative predictions of nanomechanical properties of proteins or quantum mechanical molecular properties but also reasoning over the results and correctly predicting likely mechanisms that explain distinct molecular behaviors.

BackgroundNewer 3D culturing approaches are a promising way to better mimic the in vivo tumor microenvironment and to study the interactions between the heterogeneous cell populations of glioblastoma multiforme. Like many other tumors, glioblastoma uses extracellular vesicles as an intercellular communication system to prepare surrounding tissue for invasive tumor growth. However, little is known about the effects of 3D culture on extracellular vesicles. The aim of this study was to comprehensively characterize extracellular vesicles in 3D organoid models and compare them to conventional 2D cell culture systems.MethodsPrimary glioblastoma cells were cultured as 2D and 3D organoid models. Extracellular vesicles were obtained by precipitation and immunoaffinity, with the latter allowing targeted isolation of the CD9/CD63/CD81 vesicle subpopulation. Comprehensive vesicle characterization was performed and miRNA expression profiles were generated by smallRNA-sequencing. In silico analysis of differentially regulated miRNAs was performed to identify mRNA targets and corresponding signaling pathways. The tumor cell media and extracellular vesicle proteome were analyzed by high-resolution mass spectrometry.ResultsWe observed an increased concentration of extracellular vesicles in 3D organoid cultures. Differential gene expression analysis further revealed the regulation of twelve miRNAs in 3D tumor organoid cultures (with nine miRNAs down and three miRNAs upregulated). MiR-23a-3p, known to be involved in glioblastoma invasion, was significantly increased in 3D. MiR-7-5p, which counteracts glioblastoma malignancy, was significantly decreased. Moreover, we identified four miRNAs (miR-323a-3p, miR-382-5p, miR-370-3p, miR-134-5p) located within the DLK1-DIO3 domain, a cancer-associated genomic region, suggesting a possible importance of this region in glioblastoma progression. Overrepresentation analysis identified alterations of extracellular vesicle cargo in 3D organoids, including representation of several miRNA targets and proteins primarily implicated in the immune response.ConclusionOur results show that 3D glioblastoma organoid models secrete extracellular vesicles with an altered cargo compared to corresponding conventional 2D cultures. Extracellular vesicles from 3D cultures were found to contain signaling molecules associated with the immune regulatory signaling pathways and as such could potentially change the surrounding microenvironment towards tumor progression and immunosuppressive conditions. These findings suggest the use of 3D glioblastoma models for further clinical biomarker studies as well as investigation of new therapeutic options.

Consumption of certain probiotic strains may be beneficial for reducing the risk of acute upper respiratory tract infections (URTIs), however, underlying immunological mechanisms are elusive. Bifidobacterium lactis Bl-04™ has been reported in humans to significantly reduce the risk of URTIs, affect the innate immunity in the nasal mucosa, and reduce nasal lavage virus titer after a rhinovirus (RV) challenge. To study the immunological mechanisms, we investigated the effect of Bl-04 on cytokine production and transcriptomes of human monocyte-derived macrophages (Mfs) and dendritic cells (DCs), and further on RV replication and cytokine production in MRC-5 fibroblasts. The results showed that Bl-04 modulates antiviral immune responses and potentiates cytokine production during viral challenge mimic in immune cells. However, effect of Bl-04 on RV replication and cytokine production in fibroblasts was negligible. Overall, the findings suggest that Bl-04 mildly stimulates antiviral immunity in Mfs and DCs, and potentially influences viral replication in fibroblasts that however warrants further investigations.

Abstract TSPO is a promising novel tracer target for positron-emission tomography (PET) imaging of brain tumors. However, due to the heterogeneity of cell populations that contribute to the TSPO-PET signal, imaging interpretation may be challenging. We therefore evaluated TSPO enrichment/expression in connection with its underlying histopathological and molecular features in gliomas. We analyzed TSPO expression and its regulatory mechanisms in large in silico datasets and by performing direct bisulfite sequencing of the TSPO promotor. In glioblastoma tissue samples of our TSPO-PET imaging study cohort, we dissected the association of TSPO tracer enrichment and protein labeling with the expression of cell lineage markers by immunohistochemistry and fluorescence multiplex stains. Furthermore, we identified relevant TSPO-associated signaling pathways by RNA sequencing. We found that TSPO expression is associated with prognostically unfavorable glioma phenotypes and that TSPO promotor hypermethylation is linked to IDH mutation. Careful histological analysis revealed that TSPO immunohistochemistry correlates with the TSPO-PET signal and that TSPO is expressed by diverse cell populations. While tumor core areas are the major contributor to the overall TSPO signal, TSPO signals in the tumor rim are mainly driven by CD68-positive microglia/macrophages. Molecularly, high TSPO expression marks prognostically unfavorable glioblastoma cell subpopulations characterized by an enrichment of mesenchymal gene sets and higher amounts of tumor-associated macrophages. In conclusion, our study improves the understanding of TSPO as an imaging marker in gliomas by unveiling IDH-dependent differences in TSPO expression/regulation, regional heterogeneity of the TSPO PET signal and functional implications of TSPO in terms of tumor immune cell interactions.

Abstract Background Disruption of glucocorticoid (GC) signaling in osteoblasts results in a marked attenuation of acute antibody-induced arthritis. The role of endogenous GCs in chronic inflammatory arthritis is however not fully understood. Here, we investigated the impact of endogenous GC signaling in osteoblasts on inflammation and bone integrity under chronic inflammatory arthritis by inactivating osteoblastic GC signaling in a long-term K/BxN serum transfer-induced induced arthritis (STIA) model. Methods Intracellular GC signaling in osteoblasts was disrupted by transgenic (tg) overexpression of 11beta-hydroxysteroid dehydrogenase type 2 (11ß-HSD2). Inflammatory arthritis was induced in 5-week-old male tg mice and their wild type (WT) littermates by intraperitoneal (i.p.) injection of K/BxN serum while controls (CTRLs) received phosphate-buffered saline (PBS). In a first cohort, K/BxN STIA was allowed to abate until the endpoint of 42 days (STIA). To mimic rheumatic flares, a second cohort was additionally injected on days 14 and 28 with K/BxN serum (STIA boost). Arthritis severity was assessed daily by clinical scoring and ankle size measurements. Ankle joints were assessed histopathologically. Systemic effects of inflammation on long bone metabolism were analyzed in proximal tibiae by micro-computed tomography (μCT) and histomorphometry. Results Acute arthritis developed in both tg and WT mice (STIA and STIA boost) and peaked around day 8. While WT STIA and tg STIA mice showed a steady decline of inflammation until day 42, WT STIA boost and tg STIA boost mice exhibited an arthritic phenotype over a period of 42 days. Clinical arthritis severity did not differ significantly between WT and tg mice, neither in the STIA nor in the STIA boost cohorts. Correspondingly, histological indices of inflammation, cartilage damage, and bone erosion showed no significant difference between WT and tg mice on day 42. Histomorphometry revealed an increased bone turnover in tg CTRL and tg STIA boost compared to WT CTRL and WT STIA boost animals, respectively. Conclusions In contrast to the previously reported modulating effects of endogenous GC signaling in osteoblasts during acute K/BxN STIA, this effect seems to perish during the chronic inflammatory and resolution phase. These findings indicate that endogenous GC signaling in osteoblasts may mainly be relevant during acute and subacute inflammatory processes.

Abstract Background Sex-specific physical and mental functioning trajectory classification could offer a way of understanding the differences in healthcare use at older age. Methods Using latent growth mixture models, sex-specific physical and mental functioning trajectory classes were formed for 1991 participants (mean age 61.5 years) of the Helsinki Birth Cohort Study. Physical and mental functioning were evaluated with the SF-36 survey conducted in clinical examinations in 2001–2004, 2011–2013, and 2017–2018. First and follow-up outpatient visits, emergency visits, and hospital days were extracted from a national register between the first clinical examination and the year 2017. We used regression models to examine the associations between healthcare use and trajectory classes. Results Two physical and mental functioning trajectory classes, high and intermediate, were observed for both sexes. The intermediate physical functioning trajectory class was associated with higher utilization rates of all examined specialized healthcare services (fully-adjusted IRRs varying 1.36–1.58; 95% CI = 1.03–1.79, 95% CI = 1.21–2.05) compared to the high trajectory class. Relative to the high trajectory class, the intermediate mental trajectory class was associated with the use of first outpatient visits (fully-adjusted IRRs 1.17, 95% CI = 1.03–1.33 for men, and 1.16, 95% CI = 1.04–1.30 for women). The findings were similar among both sexes. Conclusions Compared to the high trajectory class, the intermediate physical functioning trajectory class was associated with greater specialized healthcare use and the intermediate mental trajectory class with first outpatient visits. Public health interventions should be considered to support functioning with aging.

Abstract The study of biological materials and bio‐inspired materials science is well established; however, surprisingly little knowledge is systematically translated to engineering solutions. To accelerate discovery and guide insights, an open‐source autoregressive transformer large language model (LLM), BioinspiredLLM, is reported. The model is finetuned with a corpus of over a thousand peer‐reviewed articles in the field of structural biological and bio‐inspired materials and can be prompted to recall information, assist with research tasks, and function as an engine for creativity. The model has proven that it is able to accurately recall information about biological materials and is further strengthened with enhanced reasoning ability, as well as with Retrieval‐Augmented Generation (RAG) to incorporate new data during generation that can also help to traceback sources, update the knowledge base, and connect knowledge domains. BioinspiredLLM also has shown to develop sound hypotheses regarding biological materials design and remarkably so for materials that have never been explicitly studied before. Lastly, the model shows impressive promise in collaborating with other generative artificial intelligence models in a workflow that can reshape the traditional materials design process. This collaborative generative artificial intelligence method can stimulate and enhance bio‐inspired materials design workflows. Biological materials are at a critical intersection of multiple scientific fields and models like BioinspiredLLM help to connect knowledge domains.

Background: Diet is associated with major adverse cardiovascular events (MACE). Objective: We evaluated the associations between empirically derived dietary patterns and MACE. Design: Prospective cohort study. Setting: The Concord Health and Ageing in Men Project, Sydney, Australia. Participants: 539 community-dwelling older Australian men aged 75 years and older. Methods: Men underwent dietary assessment using a validated dietitian-administered diet history questionnaire. Cox regression analyses were conducted between MACE and the three dietary patterns identified from factor analysis. Five-point MACE comprised of all-cause mortality, myocardial infarction (MI), congestive cardiac failure (CCF), coronary revascularisation, and/or ischaemic stroke. Four-point MACE included the four endpoints of MI, CCF, coronary revascularisation, and/or ischaemic stroke, and excluded all-cause mortality. Results: At a median of 5.3 (IQR 4.6–6.3) years of follow-up, the incidences were: five-point MACE 31.2% (n = 168); four-point MACE excluding all-cause mortality 17.8% (n = 96); all-cause mortality 20.1% (n = 111); CCF 11.3% (n = 61); MI 3.7% (n = 20); stroke 3.2% (n = 17); and coronary revascularisation 3.1% (n = 15). In fully adjusted analyses, compared to the bottom tertile, the middle tertile of ‘vegetables-legumes-seafood’ dietary pattern was associated with reduced five-point MACE (HR 0.67 [95% CI: 0.45, 0.99, P = .047]), and CCF (HR 0.31 [95% CI: 0.15, 0.65, P = .002]), whilst the middle tertile of ‘wholegrains-milk-other fruits’ dietary pattern was associated with increased five-point MACE (HR 1.78 [95% CI: 1.17, 2.70, P = .007]), four-point MACE (HR 1.92 [95% CI: 1.12, 3.30, P = .018]), and CCF (HR 2.33 [95% CI: 1.17, 4.65, P = .016]). For the ‘discretionary-starchy vegetables-processed meats’ dietary pattern, a higher score was associated with increased five-point MACE (HR 1.33 [95% CI: 1.09, 1.62, P = .004]), and all-cause mortality (HR 1.63 [95% CI: 1.26, 2.12, P < .001]), and compared to the bottom tertile, the top tertile was associated with increased all-cause mortality (HR 2.26 [95% CI: 1.27, 4.00, P = .005]). Conclusion: Older men may benefit from consuming a ‘vegetables-legumes-seafood’ dietary pattern rather than ‘discretionary-starchy vegetables-processed meats’ and ‘wholegrains-milk-other fruits’ dietary patterns for the prevention of MACE.

Objective: Enamel and dental biofilm might serve as alternative matrices for determination of illicit and medical drugs. Thus, this study aims at evaluating possible correlations between detected drug concentrations in the matrices and simulated drug use in situ. Design: Eleven subjects wore intraoral splints with embedded demineralized bovine enamel samples. Drug use was simulated by mouth rinsing with a 1.0 μg/ml drug solution three times daily for 1 min (study A) or by incubation of the splints in a 10 μg/ml drug solution once a day for 30 min (study B). Amphetamines, opiates, cocaine and benzoylecgonine were used as drugs. After 11 days, biofilm and enamel samples of the intraoral splints were analyzed by liquid chromatography mass spectrometry after drying and extraction via ultrasonication with acetonitrile (biofilm) or methanol (enamel). Results: In study A, median and mean drug concentration ± standard deviation were 1.3 pg/mg and 6.4 ± 11 pg/mg in biofilm and 0.2 pg/mg and 0.5 ± 0.9 pg/mg in enamel. In study B, median and mean drug concentration ± standard deviation were 350 pg/mg and 1100 ± 1600 pg/mg in biofilm and 5.8 pg/mg and 9.9 ± 10 pg/mg in enamel. Conclusions: Overall, there were considerable interindividual concentration differences. Correlations between concentrations in the two sample materials were shown. The results of this pilot study revealed a dependence of concentrations on intensity and duration of drug contact. Thus, important information on past drug use might be provided in forensic cases by analysis of dental biofilm and enamel.

Objectives: U.S. and World Health Organization Selected Practice Recommendations for Contraceptive Use state people may have an advanced supply of emergency contraception (EC) to minimize treatment delays. We sought to characterize the potential improvement in effectiveness of 1.5 mg levonorgestrel (LNG-EC) if it were taken up to a few hours before unprotected sex. Study design: We expanded on an existing mathematical model for the maximum attainable effectiveness of LNG-EC, assuming it exclusively works to disrupt ovulation, and compared results with point estimates from nine studies when it was taken up to 72 hours after sex. We then modelled how effectiveness might have improved if subjects had taken LNG-EC up to 3 hours before sex. Results: Taking LNG-EC immediately after sex could potentially reduce the risk of unintended pregnancy by 91%. However, population-average maximum attainable effectiveness levels ranged from just 49% to 67% when accounting for the distributions of postcoital treatment delays in the example studies. If half the subjects had taken it 3 hours before sex, then maximum effectiveness levels would have ranged from 70% to 81%. Conclusions: At the individual level, taking LNG-EC a few hours before sex is a logical extension of Selected Practice Recommendations regarding an advanced supply of EC and, based on our modeling, should be advocated for people who can reasonably anticipate an unprotected sex act. In the absence of more clinical data, however, people should not routinely rely on precoital use of LNG-EC to prevent pregnancy unless modern, effective contraceptives are inaccessible to them. Implications: Based on mathematical modeling, individuals who anticipate needing to take LNG-EC for an impending unprotected act of sex could further reduce their chance of an undesired pregnancy by taking it a few hours in advance.

Background: Diet may be associated with frailty. Objective: We aimed to evaluate the associations between empirically derived dietary patterns and frailty in older men. Design: Prospective cohort study. Setting: The Concord Health and Ageing in Men Project, Sydney, Australia. Participants: 785 community-dwelling older Australian men aged 75 years and older. Methods: Men underwent dietary assessment using a validated dietitian-administered diet history questionnaire. Factor analysis identified three dietary patterns. Multinomial logistic regression was conducted between frailty and dietary patterns for cross-sectional analyses and longitudinal analyses over a 3-year follow-up. Frailty was defined by the Fried frailty phenotype. Results: Of the 785 men, pre-frailty was prevalent in 47.1% (n = 370), and frailty in 8.3% (n = 65). In fully adjusted cross-sectional analyses, the top tertile and a higher ‘vegetables-legumes-seafood’ dietary pattern score were associated with reduced prevalence of frailty (OR 0.34 [95% CI: 0.12, 0.93, P = .036]) and OR 0.50 [95% CI: 0.30, 0.83, P = .007] respectively). The top tertile of the ‘discretionary-starchy vegetables-processed meats’ dietary pattern was also associated cross-sectionally with increased prevalence of pre-frailty (OR 1.75 [95% CI: 1.08, 2.83, P = .022]). Of the 296 robust men in fully adjusted longitudinal analyses, the incidence of pre-frailty was 52.4% (n = 155), and frailty was 5.4% (n = 16) over a 3-year follow-up. The middle tertile of the ‘vegetables-legumes-seafood’ dietary pattern had a non-significant trend towards reduced incident pre-frailty (OR 0.52 [95% CI: 0.27, 1.00, P = .050]). Conclusion: Consumption of a ‘vegetables-legumes-seafood’ dietary pattern appears to be less favoured by frail older men.

Abstract Glioblastoma (GB) IDH-wildtype is the most malignant primary brain tumor. It is particularly resistant to current immunotherapies. Translocator protein 18 kDa (TSPO) is upregulated in GB and correlates with malignancy and poor prognosis, but also with increased immune infiltration. Here, we studied the role of TSPO in the regulation of immune resistance of human GB cells. The role of TSPO in tumor immune resistance was experimentally determined in primary brain tumor initiating cells (BTICs) and cell lines through genetic manipulation of TSPO expression and subsequent cocultures with antigen specific cytotoxic T cells and autologous tumor-infiltrating T cells. Death inducing intrinsic and extrinsic apoptotic pathways affected by TSPO were investigated. TSPO-regulated genes mediating apoptosis resistance in BTICs were identified through gene expression analysis and subsequent functional analyses. TSPO transcription in primary GB cells correlated with CD8+ T cell infiltration, cytotoxic activity of T cell infiltrate, expression of TNFR and IFNGR and with the activity of their downstream signalling pathways, as well as with the expression of TRAIL receptors. Coculture of BTICs with tumor reactive cytotoxic T cells or with T cell-derived factors induced TSPO up-regulation through T cell derived TNFα and IFNγ. Silencing of TSPO sensitized BTICs against T cell-mediated cytotoxicity. TSPO selectively protected BTICs against TRAIL-induced apoptosis by regulating apoptosis pathways. TSPO also regulated the expression of multiple genes associated with resistance against apoptosis. We conclude that TSPO expression in GB is induced through T cell-derived cytokines TNFα and IFNγ and that TSPO expression protects GB cells against cytotoxic T cell attack through TRAIL. Our data thereby provide an indication that therapeutic targeting of TSPO may be a suitable approach to sensitize GB to immune cell-mediated cytotoxicity by circumventing tumor intrinsic TRAIL resistance.

We report the case of a 69-year-old female with systemic mastocytosis, diagnosed based on widespread pigmented papules and macules, elevated serum tryptase levels and confirmatory skin and bone marrow biopsy, on a background of osteoporosis. A CT demonstrated multiple sclerotic lesions within lumbar vertebral bodies, sacrum and ileum, with surrounding osteolysis but no obvious compression fractures. She was treated with the RANK-L inhibitor denosumab, resulting in significant bone mineral density gain over the following 5 years. However, her serum tryptase levels gradually increased during this period despite treatment with the multikinase inhibitor, midostaurin. It is thus conceivable that her rapid increase in bone mineral density may be partly contributed by a predominance of pro-osteoblastic mediators released by abnormal mast cells, suggestive of more advanced disease. This case highlights the complexities of systemic mastocytosis-related bone disease and the interplay of numerous mediators contributing to a phenotype of both increased bone resorption and formation.

Abstract Multi-omics datasets are becoming of key importance to drive discovery in fundamental research as much as generating knowledge for applied biotechnology. However, the construction of such large datasets is usually time-consuming and expensive. Automation might enable to overcome these issues by streamlining workflows from sample generation to data analysis. Here, we describe the construction of a complex workflow for the generation of high-throughput microbial multi-omics datasets. The workflow comprises a custom-built platform for automated cultivation and sampling of microbes, sample preparation protocols, analytical methods for sample analysis and automated scripts for raw data processing. We demonstrate possibilities and limitations of such workflow in generating data for three biotechnologically relevant model organisms, namely Escherichia coli, Saccharomyces cerevisiae, and Pseudomonas putida.

Abstract Architected materials can achieve enhanced properties compared to their plain counterparts. Specific architecting serves as a powerful design lever to achieve targeted behavior without changing the base material. Thus, the connection between architected structure and resultant properties remains an open field of great interest to many fields, from aerospace to civil to automotive applications. Here, we focus on properties related to mechanical compression, and design hierarchical honeycomb structures to meet specific values of stiffness and compressive stress. To do so, we employ a combination of techniques in a singular workflow, starting with molecular dynamics simulation of the forward design problem, augmenting with data-driven artificial intelligence models to address the inverse design problem, and verifying the behavior of de novo structures with experimentation of additively manufactured samples. We thereby demonstrate an approach for architected design that is generalizable to multiple material properties and agnostic to the identity of the base material.

The optimal body mass hypothesis posits that the body reserves of wintering birds are balanced between the risk of starvation and predation. In this study, we tested whether the body mass of wintering Great Tits (Parus major) was higher under conditions of less predictable food resources. We compared body mass, body mass index, the speed at take-off, and apparent survival of Great Tit adult males wintering in small urban areas either near feeders providing permanent access to food for months or near feeders providing irregular access to food. Body mass and body mass index were greater, while take-off speed and apparent survival were lower, in birds wintering near permanent feeders than birds wintering near irregular feeders. Thus, urban birds, with their predictable access to high energy food, did not follow the fattening strategy predicted by the optimal body mass hypothesis. This study shows that regular excess amounts of high-energy food may affect urban birds’ physiological and behavioral strategies in a non-adaptive way. We recommend irregular feeding of wintering birds and the placing of feeders in places that are safe against attacking predators.

Abstract As the resident parenchymal myeloid population in the central nervous system (CNS), microglia are strategically positioned to respond to neurotropic virus invasion and have been implicated in promoting both disease resolution and progression in the acute and post-infectious phase of virus encephalitis. In a mouse model of West Nile virus encephalitis (WNE), infection of the CNS results in recruitment of large numbers of peripheral immune cells into the brain, the majority being nitric oxide (NO)-producing Ly6Chi inflammatory monocyte-derived cells (MCs). In this model, these cells enhance immunopathology and mortality. However, the contribution of microglia to this response is currently undefined. Here we used a combination of experimental tools, including single-cell RNA sequencing (scRNA-seq), microglia and MC depletion reagents, high-dimensional spectral cytometry and computational algorithms to dissect the differential contribution of microglia and MCs to the anti-viral immune response in severe neuroinflammation seen in WNE. Intriguingly, analysis of scRNA-seq data revealed 6 unique microglia and 3 unique MC clusters that were predominantly timepoint-specific, demonstrating substantial transcriptional adaptation with disease progression over the course of WNE. While microglia and MC adopted unique gene expression profiles, gene ontology enrichment analysis, coupled with microglia and MC depletion studies, demonstrated a role for both of these cells in the trafficking of peripheral immune cells into the CNS, T cell responses and viral clearance. Over the course of infection, microglia transitioned from a homeostatic to an anti-viral and then into an immune cell-recruiting phenotype. Conversely, MC adopted antigen-presenting, immune cell-recruiting and NO-producing phenotypes, which all had anti-viral function. Overall, this study defines for the first time the single-cell transcriptomic responses of microglia and MCs over the course of WNE, demonstrating both protective and pathological roles of these cells that could potentially be targeted for differential therapeutic intervention to dampen immune-mediated pathology, while maintaining viral clearance functions.

Abstract Background The impact of the COVID-19 pandemic and consequent public health restrictions on the mental health of adolescents is of global concern. The purpose of this study was to examine how Canadian adolescents coped during the early pandemic and whether different coping methods were associated with changes in mental health from before the pandemic to the early lockdown response. Methods Using two-year linked survey data (2018–2020) from a prospective cohort of secondary school students (n = 3,577), linear regression models were used to examine whether changes in mental health (anxiety [Generalized Anxiety Disorder-7 scale], depression [Center for Epidemiologic Studies Depression 10-item scale Revised], emotion regulation [Difficulties in Emotion Regulation Scale], psychosocial well-being [Flourishing scale]) were related to each coping behaviour. Results The most common reported coping behaviours included staying connected with friends online (78.8%), playing video games, watching TV/movies, and/or surfing the internet/social media (76.2%), studying or working on schoolwork (71.0%), and getting exercise (65.2%). The use of positive coping mechanisms during the early pandemic period (e.g., keeping a regular schedule, time with family, time with friends online) was associated with less adverse mental health changes from before to during the early lockdown; whereas, negative coping mechanisms (e.g., spending time alone, eating junk food) were consistently associated with more adverse mental health changes. Conclusion This study demonstrates the importance of social support and connections with both friends and family, as well as keeping and maintaining a routine, over the pandemic. Interventions supporting positive relationships and engagement in these coping behaviours may be protective for adolescent mental health during disruptive events.