Explore the vast range of titles available.

Embryonic stem cells can be incorporated into the developing embryo and its germ line, but, when cultured alone, their ability to generate embryonic structures is restricted. They can interact with trophoblast stem cells to generate structures that break symmetry and specify mesoderm, but their development is limited as the epithelial–mesenchymal transition of gastrulation cannot occur. Here, we describe a system that allows assembly of mouse embryonic, trophoblast and extra-embryonic endoderm stem cells into structures that acquire the embryo’s architecture with all distinct embryonic and extra-embryonic compartments. Strikingly, such embryo-like structures develop to undertake the epithelial–mesenchymal transition, leading to mesoderm and then definitive endoderm specification. Spatial transcriptomic analyses demonstrate that these morphological transformations are underpinned by gene expression patterns characteristic of gastrulating embryos. This demonstrates the remarkable ability of three stem cell types to self-assemble in vitro into gastrulating embryo-like structures undertaking spatio-temporal events of the gastrulating mammalian embryo. Sozen et al. devise an approach to combine embryonic stem cells, trophoblast stem cells and extra-embryonic endoderm stem cells into self-assembling embryo-like structures, which recapitulate key hallmarks of gastrulation in vitro.

In the version of this Technical Report originally published, the competing interests statement was missing. The authors declare no competing interests; this statement has now been added in all online versions of the Report.

Superovulation is often used to increase the number of oocytes that can be collected from donor females for in vitro fertilization. Donor age can affect the quantity and quality of oocytes produced during superovulation, and in some strains of mice juvenile females are optimal donors. The authors reviewed donor and oocyte records from a breeding program to evaluate how donor age affects the number and fertilization efficiency of oocytes collected from C57BL/6J mice. Generally fewer oocytes per donor were collected from females aged > 32 d than from females aged 21–32 d. Fertilization efficiency of oocytes generally declined with donor age when oocytes were fertilized with fresh or with stored sperm. These findings suggest that the use of younger C57BL/6J donors, instead of older donors, can reduce the number of donors needed for IVF procedures.

Heart failure remains a leading cause of morbidity and mortality worldwide. Suitable in vitro models to accurately replicate the pathological environment in heart failure with reduced and preserved ejection fraction (HFrEF/HFpEF) are limited, hampering mechanistic studies and drug screening. In particular, these models rarely incorporate immune cells, which play a critical role in heart failure. To address these limitations, we developed an isogenic 3D induced pluripotent stem cell (iPSC)-derived cardiac spheroid model incorporating monocytes. Cardiac spheroids were assembled from three healthy female iPSC lines: three-cell-type (3CT) spheroids consisting of iPSC-derived cardiomyocytes, cardiac fibroblasts, and endothelial cells, and four-cell-type (4CT) spheroids additionally containing monocytes. After six days of culture, established spheroids were treated for 24 h with different known heart failure-associated triggers (glucose & tumour necrosis factor alpha (TNFα) or ischaemia with/without reoxygenation). Differences between treated and control 3CT and 4CT spheroids were investigated at the cellular, molecular, and functional levels using confocal microscopy, RNA expression (qPCR and RNA sequencing), protein secretion using proximity extension assay technology (Olink), and functional analyses of beating rate, contraction, and relaxation. The results confirmed successful monocyte integration in 4CT spheroids, and only spheroids with monocytes (4CTs) exhibited changes in beating rate and relaxation duration upon stimulation, highlighting the necessity of incorporating immune cells to successfully mimic heart failure-associated functional changes. Along with a more pronounced global transcriptomic treatment response and inflammatory changes, additional transcriptomic alterations previously linked to heart failure in patients, as well as changes in metabolism, ion channels, and extracellular matrix pathways, were observed in 4CT compared with 3CT spheroids. We showed that immune cell incorporation enhances the functional and transcriptional responses of engineered cardiac tissue to relevant heart failure triggers in vitro and is essential for future studies to elucidate the cellular crosstalk and pathomechanisms. Heart failure continues to be a predominant cause of morbidity and mortality, necessitating the development of innovative therapeutic strategies, particularly in light of the rising prevalence of obesity and diabetes mellitus. We introduced an isogenic in vitro spheroid model comprising iPSC-derived cardiomyocytes, cardiac fibroblasts, endothelial cells, and monocytes to examine the effects of heart failure-associated triggers on cardiac tissue. Our findings indicate that spheroids incorporating monocytes exhibit a more pronounced response to heart failure-associated triggers and demonstrate greater differential transcriptional and functional responses than spheroids lacking immune cells. This model

The East Sea (also known as the Japan Sea) is connected to the Northwest Pacific via shallow straits and has independent deep water circulation, as a model miniature ocean. The radiocarbon age of dissolved organic carbon (DOC) in the East Sea ranged from 2,000 to 3,700 years, exceeding the water turnover time (∼100 years). The oldest DOC was found in the subsurface layer characterized by the Tsushima Warm Water. Comparison of the radiocarbon content and concentration of DOC in the East Sea to those in the ocean suggests that aged DOC was transported conservatively from the Northwest Pacific to the East Sea via the shallow Tsushima Warm Current. The fractions of DOC released by serial‐oxidation of the oldest DOC sample had identical radiocarbon ages, implying that refractory DOC was produced in situ and added to the DOC pool in the East Sea. Plain Language Summary The East Sea (also known as the Japan Sea) is connected to the Northwest Pacific exchanging surface water only through shallow passages. The radiocarbon age of DOC in the East Sea is old, ranging from 2,000 to 3,700 years. This cannot be explained by DOC production and aging in the East Sea. Instead, it appears that old DOC is transported into the East Sea from the Northwest Pacific. The oldest DOC was found in the subsurface layer instead of the deepest layer. We show that this oldest DOC is a mixture of the old DOC from the North Pacific and modern DOC produced in the East Sea however their characteristics in terms of resistance to oxidation were modified. These findings provide clues for understanding the global DOC cycling especially in the surface ocean. Key Points Radiocarbon ages of dissolved organic carbon (DOC) in the East Sea (Japan Sea) ranged from 2,000 to 3,700 years, exceeding the water turnover time The oldest DOC at subsurface depths in the East Sea was transported from the North Pacific via the Tsushima Warm Current The aged DOC from the North Pacific is mixed with younger DOC produced in situ and spreads to the deep waters in the East Sea

The Viviparidae, commonly known as River Snails, is a dominant group of freshwater snails with a nearly worldwide distribution that reaches its highest taxonomic and morphological diversity in Southeast Asia. The rich fossil record is indicative of a probable Middle Jurassic origin on the Laurasian supercontinent where the group started to diversify during the Cretaceous. However, it remains uncertain when and howthe biodiversity hotspot in Southeast Asiawas formed. Here,we used a comprehensive genetic data set containing both mitochondrial and nuclear markers and comprising species representing 24 out of 28 genera from throughout the range of the family. To reconstruct the spatiotemporal evolution of viviparids on a global scale, we reconstructed a fossil-calibrated phylogeny. We further assessed the roles of cladogenetic and anagenetic events in range evolution. Finally, we reconstructed the evolution of shell features by estimating ancestral character states to assess whether the appearance of sculptured shell morphologies was driven by major habitat shifts. The molecular phylogeny supports the monophyly of the three subfamilies, the Bellamyinae, Lioplacinae, and Viviparinae, but challenges the currently accepted genus-level classification in several cases. The almost global distribution of River Snails has been influenced both by comparatively ancient vicariance and more recent founder events. In Southeast Asia, Miocene dispersal was a main factor in shaping the modern species distributions. A recurrent theme across different viviparid taxa is that many species living in lentic waters exhibit sculptured shells, whereas only one strongly sculptured species is known from lotic environments. We show that such shell sculpture is habitat-dependent and indeed evolved several times independently in lentic River Snails. Considerably high transition rates between shell types in lentic habitats probably caused the co-occurrence of morphologically distinct shell types in several lakes. In contrast, directional evolution toward smooth shells in lotic habitats, as identified in the present analyses, explains why sculptured shells are rarely found in these habitats. However, the specific factors that promoted changes in shell morphology require further work.

Isolated rapid-eye-movement (REM) sleep behaviour disorder (IRBD) can be part of the prodromal stage of the α-synucleinopathies Parkinson's disease and dementia with Lewy bodies. Real-time quaking-induced conversion (RT-QuIC) analysis of CSF has high sensitivity and specificity for the detection of misfolded α-synuclein in patients with Parkinson's disease and dementia with Lewy bodies. We investigated whether RT-QuIC could detect α-synuclein in the CSF of patients with IRBD and be used as a biomarker of prodromal α-synucleinopathy. In this longitudinal observational study, CSF samples were obtained by lumbar puncture from patients with video polysomnography-confirmed IRBD recruited at a specialised sleep disorders centre in Barcelona, Spain, and from controls free of neurological disease. CSF samples were stored until analysed using RT-QuIC. After lumbar puncture, participants were assessed clinically for neurological status every 3–12 months. Rates of neurological disease-free survival were estimated using the Kaplan-Meier method. Disease-free survival rates were assessed from the date of lumbar puncture to the date of diagnosis of any neurodegenerative disease, or to the last follow-up visit for censored observations. 52 patients with IRBD and 40 healthy controls matched for age (p=0·20), sex (p=0·15), and duration of follow-up (p=0·27) underwent lumbar puncture between March 23, 2008, and July 16, 2017. The CSF α-synuclein RT-QuIC assay was positive in 47 (90%) patients with IRBD and in four (10%) controls, resulting in a sensitivity of 90·4% (95% CI 79·4–95·8) and a specificity of 90·0% (95% CI 76·9–96·0). Mean follow-up from lumbar puncture until the end of the study (July 31, 2020) was 7·1 years (SD 2·8) in patients with IRBD and 7·7 years (2·9) in controls. During follow-up, 32 (62%) patients were diagnosed with Parkinson's disease or dementia with Lewy bodies a mean 3·4 years (SD 2·6) after lumbar puncture, of whom 31 (97%) were α-synuclein positive at baseline. Kaplan-Meier analysis showed that patients with IRBD who were α-synuclein negative had lower risk for developing Parkinson's disease or dementia with Lewy bodies at 2, 4, 6, 8, and 10 years of follow-up than patients with IRBD who were α-synuclein positive (log-rank test p=0·028; hazard ratio 0·143, 95% CI 0·019–1·063). During follow-up, none of the controls developed an α-synucleinopathy. Kaplan-Meier analysis showed that participants who were α-synuclein negative (ie, five patients with IRBD plus 36 controls) had lower risk of developing Parkinson's disease or dementia with Lewy bodies at 2, 4, 6, 8 and 10 years after lumbar puncture than participants who were α-synuclein positive (ie, 47 patients with IRBD plus four controls; log-rank test p<0·0001; hazard ratio 0·024, 95% CI 0·003–0·177). In patients with IRBD, RT-QuIC detects misfolded α-synuclein in the CSF with both sensitivity and specificity of 90%, and α-synuclein positivity was associated with increased risk of subsequent diagnosis of Parkinson's disease or dementia with Lewy bodies. Detection of α-synuclein in the CSF represents a potential prodromal marker of Parkinson's disease and dementia with Lewy bodies. If these findings are replicated in additional cohorts, detection of CSF α-synuclein by RT-QuIC could be used to enrich IRBD cohorts in neuroprotective trials, particularly when assessing interventions that target α-synuclein. Department of Health and Social Care Policy Research Programme, the Scottish Government, and the Weston Brain Institute.

INTRODUCTION:Canada has a high burden of inflammatory bowel disease (IBD). Historical trends of IBD incidence and prevalence were analyzed to forecast the Canadian burden over the next decade.METHODS:Population-based surveillance cohorts in 8 provinces derived from health administrative data assessed the national incidence (2007-2014) and prevalence (2002-2014) of IBD. Autoregressive integrated moving average models were used to forecast incidence and prevalence, stratified by age, with 95% prediction intervals (PI), to 2035. The average annual percentage change (AAPC) with 95% confidence interval (CI) was calculated for the forecasted incidence and prevalence.RESULTS:The national incidence of IBD is estimated to be 29.9 per 100,000 (95% PI 28.3-31.5) in 2023. With a stable AAPC of 0.36% (95% CI −0.05 to 0.72), the incidence of IBD is forecasted to be 31.2 per 100,000 (95% PI 28.1-34.3) in 2035. The incidence in pediatric patients (younger than 18 years) is increasing (AAPC 1.27%; 95% CI 0.82-1.67), but it is stable in adults (AAPC 0.26%; 95% CI −0.42 to 0.82). The prevalence of IBD in Canada was 843 per 100,000 (95% PI 716-735) in 2023 and is expected to steadily climb (AAPC 2.43%; 95% CI 2.32-2.54) to 1,098 per 100,000 (95% PI 1,068-1,127) by 2035. The highest prevalence is in seniors with IBD (1,174 per 100,000 in 2023; AAPC 2.78%; 95% CI 2.75-2.81).DISCUSSION:Over the next decade, the Canadian health care systems will contend with the juxtaposition of rising incidence of pediatric IBD and a rising prevalence of overall IBD driven by the aging population.

Background: Drought is currently a global environmental problem, which inhibits plant growth and development and seriously restricts crop yields. Many plants exposed to drought stress can generate stress memory, which provides some advantages for resisting recurrent drought. DNA methylation is a mechanism involved in stress memory formation, and many plants can alter methylation levels to form stress memories; however, it remains unclear whether Medicago ruthenica exhibits drought stress memory, as the epigenetic molecular mechanisms underlying this process have not been described in this species. Methods: We conducted methylome and transcriptome sequencing to identify gene methylation and expression changes in plants with a history of two drought stress exposures. Results: Methylation analysis showed that drought stress resulted in an approximately 4.41% decrease in M. ruthenica genome methylation levels. The highest methylation levels were in CG dinucleotide contexts, followed by CHG contexts, with CHH contexts having the lowest levels. Analysis of associations between methylation and transcript levels showed that most DNA methylation was negatively correlated with gene expression except methylation within CHH motifs in gene promoter regions. Genes were divided into four categories according to the relationship between methylation and gene expression; the up-regulation of hypo-methylated gene expression accounted for the vast majority (692 genes) and included genes encoding factors key for abscisic acid (ABA) and proline synthesis. The hypo-methylation of the promoter and body regions of these two gene groups induced increased gene transcription levels. Conclusions: In conclusion, DNA methylation may contribute to drought stress memory formation and maintenance in M. ruthenica by increasing the transcription levels of genes key for ABA and proline biosynthesis.

Triple-negative breast cancer (TNBC) patients have a poor prognosis and few treatment options. Mouse models of TNBC are important for development of new therapies, however, few mouse models represent the complexity of TNBC. Here, we develop a female TNBC murine model by mimicking two common TNBC mutations with high co-occurrence: amplification of the oncogene MYC and deletion of the tumor suppressor PTEN . This Myc;Ptenfl model develops heterogeneous triple-negative mammary tumors that display histological and molecular features commonly found in human TNBC. Our research involves deep molecular and spatial analyses on Myc;Ptenfl tumors including bulk and single-cell RNA-sequencing, and multiplex tissue-imaging. Through comparison with human TNBC, we demonstrate that this genetic mouse model develops mammary tumors with differential survival and therapeutic responses that closely resemble the inter- and intra-tumoral and microenvironmental heterogeneity of human TNBC, providing a pre-clinical tool for assessing the spectrum of patient TNBC biology and drug response. Few mouse models recapitulate the complexity of triple negative breast cancer (TNBC). Here, the authors develop and characterise a TNBC mouse model harbouring two common TNBC mutations: amplification of the oncogene MYC and deletion of the tumour suppressor PTEN.