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Heterozygous variants in SLC6A1, encoding the GAT-1 GABA transporter, are associated with seizures, developmental delay, and autism. The majority of affected individuals carry missense variants, many of which are recurrent germline de novo mutations, raising the possibility of gain-of-function or dominant-negative effects. To understand the functional consequences, we performed an in vitro GABA uptake assay for 213 unique variants, including 24 control variants. De novo variants consistently resulted in a decrease in GABA uptake, in keeping with haploinsufficiency underlying all neurodevelopmental phenotypes. Where present, ClinVar pathogenicity reports correlated well with GABA uptake data; the functional data can inform future reports for the remaining 72% of unscored variants. Surface localization was assessed for 86 variants; two-thirds of loss-of-function missense variants prevented GAT-1 from being present on the membrane while GAT-1 was on the surface but with reduced activity for the remaining third. Surprisingly, recurrent de novo missense variants showed moderate loss-of-function effects that reduced GABA uptake with no evidence for dominant-negative or gain-of-function effects. Using linear regression across multiple missense severity scores to extrapolate the functional data to all potential SLC6A1 missense variants, we observe an abundance of GAT-1 residues that are sensitive to substitution. The extent of this missense vulnerability accounts for the clinically observed missense enrichment; overlap with hypermutable CpG sites accounts for the recurrent missense variants. Strategies to increase the expression of the wild-type SLC6A1 allele are likely to be beneficial across neurodevelopmental disorders, though the developmental stage and extent of required rescue remain unknown.

PURPOSE: Rexinoids, agonists of nuclear retinoid X receptor (RXR), have been used for the treatment of cancers and are well tolerated in both animals and humans. However, the usefulness of rexinoids in treatment of breast cancer remains unknown. This study examines the efficacy of IRX4204, a highly specific rexinoid, in breast cancer cell lines and preclinical models to identify a biomarker for response and potential mechanism of action. EXPERIMENTAL DESIGN: IRX4204 effects on breast cancer cell growth and viability were determined using cell lines, syngeneic mouse models, and primary patient-derived xenograft (PDX) tumors. In vitro assays of cell cycle, apoptosis, senescence, and lipid metabolism were used to uncover a potential mechanism of action. Standard anti-HER2 therapies were screened in combination with IRX4204 on a panel of breast cancer cell lines to determine drug synergy. RESULTS: IRX4204 significantly inhibits the growth of HER2-positive breast cancer cell lines, including trastuzumab and lapatinib-resistant JIMT-1 and HCC1954. Treatment with IRX4204 reduced tumor growth rate in the MMTV-ErbB2 mouse and HER2-positive PDX model by 49% and 44%, respectively. Mechanistic studies revealed IRX4204 modulates lipid metabolism and induces senescence of HER2-positive cells. In addition, IRX4204 demonstrates additivity and synergy with HER2-targeted mAbs, tyrosine kinase inhibitors, and antibody-drug conjugates. CONCLUSIONS: These findings identify HER2 as a biomarker for IRX4204 treatment response and demonstrate a novel use of RXR agonists to synergize with current anti-HER2 therapies. Furthermore, our results suggest that RXR agonists can be useful for the treatment of anti-HER2 resistant and metastatic HER2-positive breast cancer.

Many autism spectrum disorder (ASD)-associated genes act as transcriptional regulators (TRs). Chromatin immunoprecipitation sequencing (ChIP-seq) was used to identify the regulatory targets of ARID1B, BCL11A, FOXP1, TBR1, and TCF7L2, ASD-associated TRs in the developing human and mouse cortex. These TRs shared substantial overlap in the binding sites, especially within open chromatin. The overlap within a promoter region, 1-2,000 bp upstream of the transcription start site, was highly predictive of brain-expressed genes. This signature was observed in 96 out of 102 ASD-associated genes. In vitro CRISPRi against ARID1B and TBR1 delineated downstream convergent biology in mouse cortical cultures. After 8 days, NeuN+ and CALB+ cells were decreased, GFAP+ cells were increased, and transcriptomic signatures correlated with the postmortem brain samples from individuals with ASD. We suggest that functional convergence across five ASD-associated TRs leads to shared neurodevelopmental outcomes of haploinsufficient disruption.

Variants in cis-regulatory elements link the noncoding genome to human pathology; however, detailed analytic tools for understanding the association between cell-level brain pathology and noncoding variants are lacking. CWAS-Plus, adapted from a Python package for category-wide association testing (CWAS), enhances noncoding variant analysis by integrating both whole-genome sequencing (WGS) and user-provided functional data. With simplified parameter settings and an efficient multiple testing correction method, CWAS-Plus conducts the CWAS workflow 50 times faster than CWAS, making it more accessible and user-friendly for researchers. Here, we used a single-nuclei assay for transposase-accessible chromatin with sequencing to facilitate CWAS-guided noncoding variant analysis at cell-type-specific enhancers and promoters. Examining autism spectrum disorder WGS data (n = 7280), CWAS-Plus identified noncoding de novo variant associations in transcription factor binding sites within conserved loci. Independently, in Alzheimers disease WGS data (n = 1087), CWAS-Plus detected rare noncoding variant associations in microglia-specific regulatory elements. These findings highlight CWAS-Pluss utility in genomic disorders and scalability for processing large-scale WGS data and in multiple-testing corrections. CWAS-Plus and its user manual are available at https://github.com/joonan-lab/cwas/ and https://cwas-plus.readthedocs.io/en/latest/, respectively.

BACKGROUND: Artificial Intelligence(AI)-based solutions for Gleason grading hold promise for pathologists, while image quality inconsistency, continuous data integration needs, and limited generalizability hinder their adoption and scalability. METHODS: We present a comprehensive digital pathology workflow for AI-assisted Gleason grading. It incorporates A!MagQC (image quality control), A!HistoClouds (cloud-based annotation), Pathologist-AI Interaction (PAI) for continuous model improvement, Trained on Akoya-scanned images only, the model utilizes color augmentation and image appearance migration to address scanner variations. We evaluate it on Whole Slide Images (WSI) from another five scanners and conduct validations with pathologists to assess AI efficacy and PAI. RESULTS: Our model achieves an average F1 score of 0.80 on annotations and 0.71 Quadratic Weighted Kappa on WSIs for Akoya-scanned images. Applying our generalization solution increases the average F1 score for Gleason pattern detection from 0.73 to 0.88 on images from other scanners. The model accelerates Gleason scoring time by 43% while maintaining accuracy. Additionally, PAI improve annotation efficiency by 2.5 times and led to further improvements in model performance. CONCLUSIONS: This pipeline represents a notable advancement in AI-assisted Gleason grading for improved consistency, accuracy, and efficiency. Unlike previous methods limited by scanner specificity, our model achieves outstanding performance across diverse scanners. This improvement paves the way for its seamless integration into clinical workflows.

Apart from northern and central China, the fossil record of the latest Miocene and Early Pliocene of Asia is not well documented and the record of South China during this interval is especially poor. Shuitangba, a site in Yunnan Province, offers a rare window into the paleoenvironment of the latest Miocene in southwestern China. Over 2400 vertebrate and macrobotanical specimens have been recovered from the site. The faunal assemblage is dominated by aquatic avian taxa and many of the mammalian taxa are those that indicate densely vegetated, water-margin habitats. Pollen and carpological remains indicate a temperate to subtropical, broad-leaved/coniferous forest around standing water, with more open areas containing grasses and herbs. Analyses of clay minerals, chemical weathering, and enamel stable isotopes suggest that Shuitangba was warmer and more humid than today, possibly with more pronounced seasonality. Results of community structure analysis indicate that the Shuitangba mammalian community was different from those of other Late Miocene Chinese sites in the high proportions of aquatic-dependent mammalian taxa. While Shuitangba shared mammalian faunal elements with other Late Miocene sites in Yunnan, it was still faunally distinct. Further, Shuitangba was depauperate in its mammalian fauna, which may have been a result of the depositional setting rather than a true reflection of regional faunal diversity.

UNLABELLED: Combinations of immune checkpoint inhibitors (ICI, including anti-PD-1/PD-L1) and chemotherapy have been FDA approved for metastatic and early-stage triple-negative breast cancer (TNBC), but most patients do not benefit. B7-H4 is a B7 family ligand with proposed immunosuppressive functions being explored as a cancer immunotherapy target and may be associated with anti-PD-L1 resistance. However, little is known about its regulation and effect on immune cell function in breast cancers. We assessed murine and human breast cancer cells to identify regulation mechanisms of B7-H4 in vitro. We used an immunocompetent anti-PD-L1-sensitive orthotopic mammary cancer model and induced ectopic expression of B7-H4. We assessed therapy response and transcriptional changes at baseline and under treatment with anti-PD-L1. We observed B7-H4 was highly associated with epithelial cell status and transcription factors and found to be regulated by PI3K activity. EMT6 tumors with cell-surface B7-H4 expression were more resistant to immunotherapy. In addition, tumor-infiltrating immune cells had reduced immune activation signaling based on transcriptomic analysis. Paradoxically, in human breast cancer, B7-H4 expression was associated with survival benefit for patients with metastatic TNBC treated with carboplatin plus anti-PD-L1 and was associated with no change in response or survival for patients with early breast cancer receiving chemotherapy plus anti-PD-1. While B7-H4 induces tumor resistance to anti-PD-L1 in murine models, there are alternative mechanisms of signaling and function in human cancers. In addition, the strong correlation of B7-H4 to epithelial cell markers suggests a potential regulatory mechanism of B7-H4 independent of PD-L1. SIGNIFICANCE: This translational study confirms the association of B7-H4 expression with a cold immune microenvironment in breast cancer and offers preclinical studies demonstrating a potential role for B7-H4 in suppressing response to checkpoint therapy. However, analysis of two clinical trials with checkpoint inhibitors in the early and metastatic settings argue against B7-H4 as being a mechanism of clinical resistance to checkpoints, with clear implications for its candidacy as a therapeutic target.

We report the discovery of 15 exceptionally luminous 10 ≲ z ≲ 14 candidate galaxies discovered in the first 0.28 deg2 of JWST/NIRCam imaging from the COSMOS-Web survey. These sources span rest-frame UV magnitudes of −20.5 > M UV > −22, and thus constitute the most intrinsically luminous z ≳ 10 candidates identified by JWST to date. Selected via NIRCam imaging, deep ground-based observations corroborate their detection and help significantly constrain their photometric redshifts. We analyze their spectral energy distributions using multiple open-source codes and evaluate the probability of low-redshift solutions; we conclude that 12/15 (80%) are likely genuine z ≳ 10 sources and 3/15 (20%) likely low-redshift contaminants. Three of our z ∼ 12 candidates push the limits of early stellar mass assembly: they have estimated stellar masses ∼ 5 × 109 M ⊙, implying an effective stellar baryon fraction of ϵ ⋆ ∼ 0.2−0.5, where ϵ ⋆ ≡ M ⋆/(f b M halo). The assembly of such stellar reservoirs is made possible due to rapid, burst-driven star formation on timescales < 100 Myr where the star formation rate may far outpace the growth of the underlying dark matter halos. This is supported by the similar volume densities inferred for M ⋆ ∼ 1010 M ⊙ galaxies relative to M ⋆ ∼ 109 M ⊙—both about 10−6 Mpc−3—implying they live in halos of comparable mass. At such high redshifts, the duty cycle for starbursts would be of order unity, which could cause the observed change in the shape of the UV luminosity function from a double power law to a Schechter function at z ≈ 8. Spectroscopic redshift confirmation and ensuing constraints of their masses will be critical to understand how, and if, such early massive galaxies push the limits of galaxy formation in the Lambda cold dark matter paradigm.

INTRODUCTION: Treatments for atopic dermatitis (AD) often fail to achieve lasting disease control. In the CrisADe CONTROL phase III study (ClinicalTrials.gov: NCT04040192), participants aged ≥ 3 months with mild to moderate AD treated with once-daily (QD) crisaborole, following initial treatment success with crisaborole twice daily (BID), had longer periods of flare-free maintenance, a higher number of flare-free days, and a lower number of flares compared with those who received vehicle. The study was an exploratory analysis of data on the maintenance of response per Investigators Static Global Assessment (ISGA; ISGA score of 0 [clear] or 1 [almost clear]) during the CrisADe CONTROL study through week 52. METHODS: Exploratory endpoints were the time to ISGA response during the open-label run-in period, and the maintenance of ISGA response and the severity and duration of flares during the double-blind maintenance period. Outcomes were stratified by age (participants aged 3 months to

Multiomics has shown promise in noninvasive risk profiling and early detection of various common diseases. In the present study, in a prospective population-based cohort with ~18 years of e-health record follow-up, we investigated the incremental and combined value of genomic and gut metagenomic risk assessment compared with conventional risk factors for predicting incident coronary artery disease (CAD), type 2 diabetes (T2D), Alzheimer disease and prostate cancer. We found that polygenic risk scores (PRSs) improved prediction over conventional risk factors for all diseases. Gut microbiome scores improved predictive capacity over baseline age for CAD, T2D and prostate cancer. Integrated risk models of PRSs, gut microbiome scores and conventional risk factors achieved the highest predictive performance for all diseases studied compared with models based on conventional risk factors alone. The present study demonstrates that integrated PRSs and gut metagenomic risk models improve the predictive value over conventional risk factors for common chronic diseases.

Background: Telehealth utilization exploded during the COVID-19 pandemic, including within school-based health programs. School-based tele-behavioral health can help programs overcome barriers of access to care, but the current state and effectiveness of such programs are unknown. Methods: A scoping literature review was conducted. Studies were included if they described in-school behavioral health services delivered via telehealth for children ages 5 to 18. From the included studies, population, location, setting, intervention, telehealth modality, clinician type, and outcomes assessed were extracted. Findings: Eighteen studies met inclusion criteria. All described psychotherapy or medication management delivered by psychologists (n = 7) and/or psychiatrists (n = 11). Treatment included psychotherapy (N = 8), psychiatric consultation (N = 7), medication management (N = 4), crisis stabilization (N = 1), and caregiver education (N = 1). Eight studies provide qualitative or quantitative outcomes, with 4 examining clinical effectiveness. Conclusions: Despite limited findings in the literature, school-based tele-behavioral health is feasible, effective, and acceptable for delivery of behavioral health care to children and adolescents.

The purpose of this research is to analyse the support that lecturers need to be able to implement a blended learning approach successfully. Blended learning is now seen as an approach that can create engaging learning environments, to enhance students' self-directed learning and improve the whole learning experience. The study investigated the perceptions of thirteen lecturers, at a particular higher education institution, regarding the advantages and challenges of blended learning in South Africa. A qualitative research design was chosen for this study because it enabled the authors to explore the theme in detail. Semi-structured open-ended interviews were conducted to collect the necessary data from the lecturers, who were chosen through the use of homogenous purposeful sampling. The data collected were coded, using the induction method. This helped to reveal relevant codes, which were categorised. A literature review was conducted, in which recent research on this topic was analysed and used to correlate the findings of the field research of this study. This study reports on the actual views and experiences of the participants. The research findings and relevance for teaching in higher education institutions are discussed. The conclusion is that, for such an approach to be successful, lecturers would require support from management, more training, improved professional development, as well as reliable technology and internet connections. In addition, lecturers would require additional time to implement such an approach.

Background: We investigated the prevalence of swallowing difficulties and associated factors in people with intellectual disability. Methods: We included people aged 50+ receiving care for people with intellectual disabilities. The Dysphagia Disorder Survey (DDS) was used to assess swallowing difficulties. We determined the agreement between the DDS and swallowing difficulties in medical records. We used logistic regression analyses to explore associated factors. Results: One thousand and fifty people were included. The prevalence of swallowing difficulties was 43.8%. Swallowing difficulties were not reported in the medical records of 83.3% of these cases. Frailty (odds ratio (OR) = 4.22, 95% CI = 2.05-8.71), mobility impairment (OR = 2.50, 95% CI = 1.01-6.19), and mealtime dependency (OR = 3.05, 95% CI = 1.10-8.47) were independently associated with swallowing difficulties. Conclusion: Swallowing difficulties are prevalent in older people with intellectual disability but may be under-recognised. Frailty may be a good indicator for population-based screening for swallowing difficulties.

Decades of research suggests that supportive faculty communication improves student retention and progression; however, most communication interventions occurred at on-site universities and were implemented briefly in one or several courses. In this article, we describe the outcomes of a consistent, personalized student outreach program implemented in every course and for students' entire tenure at a fully online university. In 2019, Western Governors University launched a student support program called Learner-Centered Faculty (LCF)--a paradigm that focuses on just-in-time, personalized, student-centered faculty support at scale. We evaluated the impact of LCF on student retention and course completion rates and found that LCF-based intervention improved retention and course completion generally and was particularly beneficial for certain groups of students. In the future, we will examine the mechanisms underlying LCF's success.

The forced closure of South African schools left learners with many challenges, especially in language learning. This study aims to explore the perceptions of teachers teaching the same subject to the same grade during the COVID-19 pandemic. The purpose is to analyse learning losses and changes in the scholastic achievement of primary school learners during the COVID-19 pandemic. A qualitative research design was chosen for this study because of its exploratory nature. Semi-structured open-ended interviews rendered data on the teachers' perceptions of academic losses as a result of the "COVID-19" pandemic. Homogenous purposeful sampling and semi-structured open-ended interviews were used to gather data, which were coded using the induction method to reveal codes, categories and subcategories within participant voices. This study reports on the challenges faced by teachers and learners, which include a lack of parental support, inadequate technological knowledge, and neglect of schoolwork. The findings and their relevance to teaching post-COVID-19 to mitigate academic losses are discussed. The conclusion is that different forms of support are needed to mitigate the negative impact of the COVID-19 pandemic on learners' scholastic performance.