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This study examined parental playfulness as a mediator of the association between mothers' emotional difficulties (anxiety, depression) and their children's psychosocial adjustment (externalizing, internalizing, and prosocial behaviours). We also examined the associations between parental playfulness and mothers' supportive / unsupportive reactions to their child's negative emotions. Participants were 197 mothers of children aged 3-8 years, who completed online questionnaires. Mothers' higher levels of emotional difficulties were associated with lower levels of parental playfulness, and these were associated with children's higher levels of externalizing and internalizing symptoms, and lower levels of prosocial behaviours. Parental playfulness mediated the relationship between mothers' emotional difficulties and children's externalizing symptoms. It was also positively related with mothers' supportive reactions to children's negative emotions and negatively related with unsupportive reactions. These findings underscore the importance of parental playfulness to children's emotional adjustment, particularly to externalizing symptoms, and may provide a basis for new parenting interventions.

The alluvial material in the two PITs of the Mayo Sina in the southern part of Chad, were subjected to a geochemical analysis in order to determine the source rock lithological composition, tectonic context, degree of weathering, type of climate at the period of weathering, and cycles of sedimentation and maturity. Plotting source lithology charts with values of La/Sc, Th/Sc, La/Co, and Cr/Th indicated that the sediments originated from a felsic source rock composition. The Enrichment of LREE/HREE and a negative chondrite Eu anomaly confirm the felsic source rock composition. A passive tectonic context for the source rock of the sediments is revealed by binary charts (SiO2 versus K2O/Na2O) in conjunction with innovative discrimination plots (DF 1 and DF 2 (Arc-Rift-Col)M1, DF (A-P)M, and DF (A-P)MT). The exceptional degree of plagioclase removal and weathering is demonstrated by the results of the Plagioclase Index of Alteration (PIX) and Chemical Index of Alteration (CIX). The proportion of SiO2/Al2O3 and the new index of compositional variation (ICVnew), which exclude iron and calcium oxides from the formula, indicate that the sediments are immature to mature. Schematic representations of K2O/Na2O ratio indicate that the sediments have undergone both primary to secondary phases of sedimentation.

The advent of sub-meter resolution topographic surveying has revolutionized active fault mapping. Light detection and ranging (lidar) collected using crewed airborne laser scanning (ALS) can provide ground coverage of entire fault systems but is expensive, while Structure-from-Motion (SfM) photogrammetry from uncrewed aerial vehicles (UAVs) is popular for mapping smaller sites but cannot image beneath vegetation. Here, we present a new UAV laser scanning (ULS) system which overcomes these limitations to survey fault-related topography cost-effectively, at desirable spatial resolutions, and even beneath dense vegetation. In describing our system, data acquisition and processing workflows, we provide a practical guide for other researchers interested in developing their own ULS capabilities. We showcase ULS data collected over faults from a variety of terrain and vegetation types across the Canadian Cordillera and compare them to conventional ALS and SfM data. Due to the lower, slower UAV flights, ULS offers improved ground return density (~260 points/m2 for the capture of a paleoseismic trenching site and ~10–72 points/m2 for larger, multi-kilometer fault surveys) over conventional ALS (~3–9 points/m2) as well as better vegetation penetration than both ALS and SfM. The resulting ~20–50 cm-resolution ULS terrain models reveal fine-scale tectonic landforms that would otherwise be challenging to image.

Commensal intestinal bacteria shape our microbiome and have decisive roles in preserving host metabolic and immune homeostasis. They conspicuously impact disease development and progression, including amyloid-beta (Aβ) and alpha (α)-synuclein pathology in neurodegenerative diseases, conveying the importance of the brain–gut–microbiome axis in such conditions. However, little is known about the longitudinal microbiome landscape and its potential clinical implications in other protein misfolding disorders, such as prion disease. We investigated the microbiome architecture throughout prion disease course in mice. Fecal specimens were assessed by 16S ribosomal RNA sequencing. We report a temporal microbiome signature in prion disease and uncovered alterations in Lachnospiraceae, Ruminococcaceae, Desulfovibrionaceae, and Muribaculaceae family members in this disease. Moreover, we determined the enrichment of Bilophila, a microorganism connected to cognitive impairment, long before the clinical manifestation of disease symptoms. Based on temporal microbial abundances, several associated metabolic pathways and resulting metabolites, including short-chain fatty acids, were linked to the disease. We propose that neuroinflammatory processes relate to perturbations of the intestinal microbiome and metabolic state by an interorgan brain–gut crosstalk. Furthermore, we describe biomarkers possibly suitable for early disease diagnostics and anti-prion therapy monitoring. While our study is confined to prion disease, our discoveries might be of equivalent relevance in other proteinopathies and central nervous system pathologies.

Abstract Everolimus is widely used in patients with advanced ER‐positive, HER2‐negative breast cancer. We looked at alterations in the PIK3CA/AKT/mTOR pathway in a multicenter cohort as potential biomarkers of efficacy. Patients with advanced ER‐positive, HER2‐negative breast cancer treated with everolimus and endocrine therapy between 2012 and 2014 in two cancer centers were included. Targeted sequencing examined mutations in PIK3CA, ESR1, and AKT1 genes. An immunochemical analysis was conducted to evaluate expression of PTEN, INPP4B, STK11, p4EBP1, and pS6. We analyzed 71 patients (44 primary tumors; 27 metastatic tissues). Median age was 63 years [58–69]. All patients had heavily pretreated advanced disease. A mutation in the PIK3CA pathway was observed in 32 samples (PIK3CA exons 10 and 21 and AKT1 exon 4 in 15.5%, 24.0%, and 5.6% of samples), and in ESR1 in 5 samples (7.0%), respectively. Most samples showed cytoplasmic expression of the PIK3CA pathway proteins. Progression‐free survival was longer in patients with a pS6 or p4EBP1 histoscore ≥ median value (6.6 versus 3.7 months, p = 0.037), and in patients with a PTEN histoscore ≤ median value (7.1 versus 5.3 months, p = 0.02). Overall survival was longer in patients with pS6 ≥ 3rd quartile (27.6 versus 19.3 months, p = 0.038) and in patients with any mutation in the PIK3CA/AKT/mTOR pathway (27.6 versus 19.3 months, p = 0.011). The prognosis of patients treated with everolimus for advanced ER‐positive, HER2‐negative breast cancer appears primarily driven by molecular features associated with the activation of the PIK3CA/AKT/mTOR pathway.

As clinical trials in oncology require substantial efforts, maximizing the insights gained from them by conducting subgroup analyses is often attempted. The goal of these analyses is to identify subgroups of patients who are likely to benefit, as well as the subgroups of patients who are unlikely to benefit from the studied intervention. International guidelines occasionally include or exclude novel medications and technologies for specific subpopulations based on such analyses of pivotal trials without requiring confirmatory trials. This Perspective discusses the importance of providing a complete dataset of clinical information when reporting subgroup analyses and explains why such transparency is key for better clinical interpretation of the results and the appropriate application to clinical care, by providing examples of transparent reporting of clinical studies and examples of incomplete reporting of clinical studies.

The functionality of photoreceptors, rods, and cones is highly dependent on their outer segments (POS), a cellular compartment containing highly organized membranous structures that generate biochemical signals from incident light. While POS formation and degeneration are qualitatively assessed on microscopy images, reliable methodology for quantitative analyses is still limited. Here, we developed methods to quantify POS (QuaPOS) maturation and quality on retinal sections using automated image analyses. POS formation was examined during the development and in adulthood of wild-type mice via light microscopy (LM) and transmission electron microscopy (TEM). To quantify the number, size, shape, and fluorescence intensity of POS, retinal cryosections were immunostained for the cone POS marker S-opsin. Fluorescence images were used to train the robust classifier QuaPOS-LM based on supervised machine learning for automated image segmentation. Characteristic features of segmentation results were extracted to quantify the maturation of cone POS. Subsequently, this quantification method was applied to characterize POS degeneration in “cone photoreceptor function loss 1” mice. TEM images were used to establish the ultrastructural quantification method QuaPOS-TEM for the alignment of POS membranes. Images were analyzed using a custom-written MATLAB code to extract the orientation of membranes from the image gradient and their alignment (coherency). This analysis was used to quantify the POS morphology of wild-type and two inherited retinal degeneration (“retinal degeneration 19” and “rhodopsin knock-out”) mouse lines. Both automated analysis technologies provided robust characterization and quantification of POS based on LM or TEM images. Automated image segmentation by the classifier QuaPOS-LM and analysis of the orientation of membrane stacks by QuaPOS-TEM using fluorescent or TEM images allowed quantitative evaluation of POS formation and quality. The assessments showed an increase in POS number, volume, and membrane coherency during wild-type postnatal development, while a decrease in all three observables was detected in different retinal degeneration mouse models. All the code used for the presented analysis is open source, including example datasets to reproduce the findings. Hence, the QuaPOS quantification methods are useful for in-depth characterization of POS on retinal sections in developmental studies, for disease modeling, or after therapeutic interventions affecting photoreceptors.

The development of new molecules for the treatment of calmodulin related cardiovascular or neurodegenerative diseases is an interesting goal. In this work, we introduce a novel strategy with four main steps: (1) chemical synthesis of target molecules, (2) Förster Resonance Energy Transfer (FRET) biosensor development and in vitro biological assay of new derivatives, (3) Cheminformatics models development and in vivo activity prediction, and (4) Docking studies. This strategy is illustrated with a case study. Firstly, a series of 4-substituted Riluzole derivatives 1–3 were synthetized through a strategy that involves the construction of the 4-bromoriluzole framework and its further functionalization via palladium catalysis or organolithium chemistry. Next, a FRET biosensor for monitoring Ca2+-dependent CaM-ligands interactions has been developed and used for the in vitro assay of Riluzole derivatives. In particular, the best inhibition (80%) was observed for 4-methoxyphenylriluzole 2b. Besides, we trained and validated a new Networks Invariant, Information Fusion, Perturbation Theory, and Machine Learning (NIFPTML) model for predicting probability profiles of in vivo biological activity parameters in different regions of the brain. Next, we used this model to predict the in vivo activity of the compounds experimentally studied in vitro. Last, docking study conducted on Riluzole and its derivatives has provided valuable insights into their binding conformations with the target protein, involving calmodulin and the SK4 channel. This new combined strategy may be useful to reduce assay costs (animals, materials, time, and human resources) in the drug discovery process of calmodulin inhibitors.

Introduction Research has shown that factors related with the migratory process (such as travelling alone, living away from family, and discrimination after arrival) considerably increase the risk of mental health problems in young migrants. Moreover, they are among the most vulnerable migration groups with a high risk of social exclusion. Objectives To identify coping strategies and behavioural changes used to deal with perceived discrimination and its impact on the emotional well-being and mental health of newly arrived young migrants in Spain. Methods A subsample of 15 audio-recorded in-depth qualitative interviews were analysed from the national action-research Migrasalud project (II IN 190517 EN 162 FA 01). The interviews were transcribed, translated from Arabic to Spanish, and analysed through content analysis. Results Most participants were males (93.3%; n=14), ranging from 18 to 20 years, and from Morocco (93.3%; n=14). All participants were from foster care placements in Barcelona and arrived to Spain as minors. Newly arrived young migrants reported that they perceived themselves as being healthy before the migratory process. Adverse experiences during the journey and discrimination after arrival impacted their well-being and mental health. Specifically, they reported perceived discrimination in their daily life due to culture, language, or origin. This negatively impacted their well-being and mental health, increasing their emotional distress response and ‘undervalued or inferior’ and ‘vulnerable’ feelings about themselves. Concerning coping with discrimination, they reported using internalised coping strategies such as ‘ignoring’ or ‘not responding’ for fear of having their legal documents revoked or not obtaining them. Their behavioural changes often occurred when they perceived unfair treatment or prejudice towards their migrant status or their socioeconomics, culture or religion. These changes were motivated by being more accepted by the local community by ‘westernisation or cultural assimilation’ and by ‘creating a good image’ of oneself and its culture. Conclusions Findings establish that the cumulative experience of post-migration stressors (such as discrimination) negatively impacts their mental health and well-being in the long term. This suggests the need for specific policies and services to address this population’s effects of post-migration risk factors. Further research is needed to explore the causes and effects of perceived discrimination on mental health more closely and to develop more targeted and effective interventions. Disclosure of Interest None Declared

Cytotoxic T lymphocyte (CTL) motility is an important feature of effective CTL responses and is impaired when CTLs become exhausted, e.g. during chronic retroviral infections. A prominent T cell exhaustion marker is programmed cell death protein 1 (PD-1) and antibodies against the interaction of PD-1 and PD-ligand 1 (PD-L1) are known to improve CTL functions. However, antibody blockade affects all PD-1/PD-L1-expressing cell types, thus, the observed effects cannot be attributed selectively to CTLs. To overcome this problem, we performed CRISPR/Cas9 based knockout of the PD-1 coding gene PDCD1 in naïve Friend Retrovirus (FV)-specific CTLs. We transferred 1,000 of these cells into mice where they proliferated upon FV-infection. Using intravital two-photon microscopy we visualized CTL motility in the bone marrow and evaluated cytotoxic molecule expression by flow cytometry. Knockout of PDCD1 improved the CTL motility at 14 days post infection and enhanced the expression of cytotoxicity markers. Our data show the potential of genetic tuning of naive antiviral CTLs and might be relevant for future designs of improved T cell-mediated therapies.

IntroductionIntraoperative radiation therapy (IORT) delivers a single accelerated radiation dose to the breast tumor bed during breast-conserving surgery (BCS). The synergistic biologic effects of simultaneous surgery and radiation remain unclear. This study explores the cellular and molecular changes induced by IORT in the tumor microenvironment and its impact on the immune response modulation.MethodsPatients with hormone receptor (HR)-positive/HER2-negative, ductal carcinoma in situ (DCIS), or early-stage invasive breast carcinoma undergoing BCS with margin re-excision were included. Histopathological evaluation and RNA-sequencing in the re-excision tissue were compared between patients with IORT (n=11) vs. non-IORT (n=11).ResultsSquamous metaplasia with atypia was exclusively identified in IORT specimens (63.6%, p=0.004), mimicking DCIS. We then identified 1,662 differentially expressed genes (875 upregulated and 787 downregulated) between IORT and non-IORT samples. Gene ontology analyses showed that IORT was associated with the enrichment of several immune response pathways, such as inflammatory response, granulocyte activation, and T-cell activation (p

OBJECTIVES Over the past 2 decades, simulation-based learning has become an essential part of medical training. Simulated clinics have proven to be effective for training medical students. Even so, this learning method presents organizational and financial challenges that limit its dissemination to all medical students, especially since the COVID-19 pandemic. Simulated teleconsultation retains the advantages of interactive simulated clinics while offering concrete solutions to the challenges faced. The project aims to explore students’ perspectives on simulated teleconsultation training compared to simulated clinics in person. METHODS Ten pre-clerkship students in the Faculty of Medicine at the University of Ottawa participated in interviews following in-person and teleconsultation simulated clinic sessions. The interview guide was developed based on previous work. The questions asked concerned experience with teleconsultation, interaction with the tutor and patient, practical or logistical obstacles, educational value and feasibility. The authors evaluated the results using a thematic analysis. RESULTS The interview analysis showed that the tutor feedback received during the simulated teleconsultation was comparable to that received after the in-person simulated clinic. Although most of the students enjoy teleconsultation, they raised the challenge of carrying out physical examinations and creating a personal connection with the tutor/patient. CONCLUSION Given the circumstances of the pandemic and students’ comfort with technology, the new generation of medical students seems prepared to embrace teleconsultation. The themes identified in the analysis will enable the necessary adjustments to be made in order to optimize their teleconsultation training, an inextricable step in promoting the active offer of healthcare services.

Summary: The type VI secretion system (T6SS), a widespread protein delivery apparatus, plays a role in bacterial competition by delivering toxic effectors into neighboring cells. Identifying new T6SS effectors and deciphering the mechanism that governs their secretion remain major challenges. Here, we report two orphan antibacterial T6SS effectors in the pathogen Pantoea agglomerans (Pa). These effectors share an N-terminal domain, Pantoea type six (PIX), that defines a widespread class of polymorphic T6SS effectors in Enterobacterales. We show that the PIX domain is necessary and sufficient for T6SS-mediated effector secretion and that PIX binds to a specialized Pa VgrG protein outside its C-terminal toxic domain. Our findings underline the importance of identifying and characterizing delivery domains in polymorphic toxin classes as a tool to reveal effectors and shed light on effector delivery mechanisms.

Summary: Multiple myeloma (MM) is an incurable hematological malignancy in which MYC alterations contribute to the malignant phenotype. Nevertheless, MYC lacks therapeutic druggability. Here, we leveraged large-scale loss-of-function screens and conducted a small molecule screen to identify genes and pathways with enhanced essentiality correlated with MYC expression. We reported a specific gene dependency in glutaminase (GLS1), essential for the viability and proliferation of MYC overexpressing cells. Conversely, the analysis of isogenic models, as well as cell lines dataset (CCLE) and patient datasets, revealed GLS1 as a non-oncogenic dependency in MYC-driven cells. We functionally delineated the differential modulation of glutamine to maintain mitochondrial function and cellular biosynthesis in MYC overexpressing cells. Furthermore, we observed that pharmaceutical inhibition of NAMPT selectively affects MYC upregulated cells. We demonstrate the effectiveness of combining GLS1 and NAMPT inhibitors, suggesting that targeting glutaminolysis and NAD synthesis may be a promising strategy to target MYC-driven MM.

Since the seminal work of Cundall and Strack (1979), the Discrete Element Method (DEM) has now become accepted as a key tool amongst researchers exploring the fundamental behavior of granular materials. Along with a sustained increase in the number of publications documenting use of DEM in research, intensive development of new open-source and commercial DEM codes has taken place in the last decades. The credibility of these software packages depends on their capacity to replicate physical observations and to reproduce theoretical expressions. Researchers often calibrate DEM codes against laboratory data to gain confidence about their predictions, however, theoretical verifications at the macro and particle levels are often omitted or not explicitly documented or acknowledged. The validation of DEM codes against theoretical expressions is fundamental to guarantee reproducibility and generality of the software, and to avoid bias in more complex simulations.In this article, a dataset providing numerical simulation data along with input files is presented. The dataset relates to a series of theoretical validation approaches, previously documented in the literature, that were here applied to verify the open-source DEM code LAMMPS. The ability of LAMMPS to capture the macroscopic behaviour of granular packages is evaluated by shearing a face-center-cubic (FCC) array of monosized spheres. The calculation of particle translational/rotational motions and forces/torques is checked by considering a clump rolling down an inclined plane. Additionally, the stress-strain behavior of Toyoura sand under “drained” and “undrained” shearing is characterized by a series of LAMMPS outputs. The dataset collected from these simulations can be employed by users to benchmark new or existing DEM codes. Both the LAMMPS input scripts and the simulation results for all the cases are available in a public repository.

BackgroundThe effect of COVID-19 in pregnancy on maternal outcomes and its association with preeclampsia and gestational diabetes mellitus have been reported; however, a detailed understanding of the effects of maternal positivity, delivery mode, and perinatal practices on fetal and neonatal outcomes is urgently needed.ObjectiveTo evaluate the impact of COVID-19 on fetal and neonatal outcomes and the role of mode of delivery, breastfeeding, and early neonatal care practices on the risk of mother-to-child transmission.Study designIn this cohort study that took place from March 2020 to March 2021, involving 43 institutions in 18 countries, 2 unmatched, consecutive, unexposed women were concomitantly enrolled immediately after each infected woman was identified, at any stage of pregnancy or delivery, and at the same level of care to minimize bias. Women and neonates were followed up until hospital discharge. COVID-19 in pregnancy was determined by laboratory confirmation and/or radiological pulmonary findings or ≥2 predefined COVID-19 symptoms. The outcome measures were indices of neonatal and perinatal morbidity and mortality, neonatal positivity and its correlation with mode of delivery, breastfeeding, and hospital neonatal care practices.ResultsA total of 586 neonates born to women with COVID-19 diagnosis and 1535 neonates born to women without COVID-19 diagnosis were enrolled. Women with COVID-19 diagnosis had a higher rate of cesarean delivery (52.8% vs 38.5% for those without COVID-19 diagnosis, P14 days). Among neonates born to mothers with COVID-19 diagnosis, birth via cesarean delivery was a risk factor for testing positive for COVID-19 (odds ratio, 2.4; 95% confidence interval, 1.2-4.7), even when severity of maternal conditions was considered and after multivariable logistic analysis. In the subgroup of neonates born to women with COVID-19 diagnosis, the outcomes worsened when the neonate also tested positive, with higher rates of neonatal intensive care unit admission, fever, gastrointestinal and respiratory symptoms, and death, even after adjusting for prematurity. Breastfeeding by mothers with COVID-19 diagnosis and hospital neonatal care practices, including immediate skin-to-skin contact and rooming-in, were not associated with an increased risk of newborn positivity.ConclusionIn this multinational cohort study, COVID-19 in pregnancy was associated with increased maternal and neonatal complications. Cesarean delivery was significantly associated with newborn COVID-19 diagnosis. Vaginal delivery should be considered the safest mode of delivery if obstetrical and health conditions allow it. Mother-to-child skin-to-skin contact, rooming-in, and direct breastfeeding were not risk factors for newborn COVID-19 diagnosis, thus well-established best practices can be continued among women with COVID-19 diagnosis.

Adequate pain management is one of the biggest challenges of the modern healthcare system. Physician perception of patient subjective pain, which is crucial to pain management, is susceptible to a host of potential biases. Here we explore the timing of physicians' work as a previously unrecognized source of systematic bias in pain management. We hypothesized that during night shifts, sleep deprivation, fatigue, and stress would reduce physicians' empathy for others' pain, leading to underprescription of analgesics for patient pain relief. In study 1, 67 resident physicians, either following a night shift or not, performed empathy for pain assessment tasks and simulated patient scenarios in laboratory conditions. As predicted, following a night shift, physicians showed reduced empathy for pain. In study 2, we explored this phenomenon in medical decisions in the field. We analyzed three emergency department datasets from Israel and the United States that included discharge notes of patients arriving with pain complaints during 2013 to 2020 (n = 13,482). Across all datasets, physicians were less likely to prescribe an analgesic during night shifts (compared to daytime shifts) and prescribed fewer analgesics than generally recommended by the World Health Organization. This effect remained significant after adjusting for patient, physician, type of complaint, and emergency department characteristics. Underprescription for pain during night shifts was particularly prominent for opioids. We conclude that night shift work is an important and previously unrecognized source of bias in pain management, likely stemming from impaired perception of pain. We consider the implications for hospitals and other organizations employing night shifts.

We report on the direct search for cosmic relic neutrinos using data acquired during the first two science campaigns of the KATRIN experiment in 2019. Beta-decay electrons from a high-purity molecular tritium gas source are analyzed by a high-resolution MAC-E filter around the end point at 18.57 keV. The analysis is sensitive to a local relic neutrino overdensity ratio of η