Generation of porcine endometrial organoids and their use as a model for enhancing embryonic attachment and elongation.

In brief: Porcine endometrial organoids (EOs) were isolated and characterized, revealing distinctive features such as unique extracellular matrix formation, fusion into uterine bud-like structures, and facilitation of embryo elongation. The yield of EOs was significantly enhanced by cryopreservation medium supplemented with the rock inhibitor (Y-27632), resulting in reduced expression of apoptotic mRNAs and microRNAs. Abstract: Endometrial organoids (EOs) are acceptable models for understanding maternal-embryonic cross talk. This study was conducted to generate EOs and optimize their cryopreservation and provide coculture modeling with embryos. The endometrial tissues were used for culturing the organoids inside domes of Matrigel®. To improve the long-term storage of EOs, 10 µM ROCK inhibitor (RI) was added to the cryopreservation medium. Day 7 parthenogenetically activated embryos were cocultured with EOs or EO outgrowths, and embryonic cell numbers and embryo attachment were monitored. Spherical EOs 100-300 µm in size can be retrieved on day 7 of culture, and larger EOs, approximately 1.5 mm in diameter, can be maintained in the Matrigel® dome for 21 days. The nuclear expression of Ki67 indicates that more than 80% of EOs nuclei were proliferative. EOs exhibit unique novel characters such as formation of extracellular matrix and ability for fusion. RI increased the yield and quality of organoids after freezing or thawing. The cell number of cocultured embryos increased five-fold, and the proportion of trophoblast outgrowths increased seven-fold compared with those of control embryos. The embryos cultured with EO-conditioned medium showed a better attachment rate than the other models, and - for the first time - we report embryonic elongation. Immunofluorescence staining of the attached embryos showed CDX2 in the periphery of EOs outgrowths. The 3D assembly and cryopreservation of EOs was optimized, and EO coculture supported embryo attachment, trophoblast outgrowth, and elongation, which would provide a valuable tool for studying the intricate processes involved in porcine embryo implantation.

Glutamine Triggers Acetylation-Dependent Degradation of Glutamine Synthetase via the Thalidomide Receptor Cereblon.

Cereblon (CRBN), a substrate receptor for the cullin-RING ubiquitin ligase 4 (CRL4) complex, is a direct protein target for thalidomide teratogenicity and antitumor activity of immunomodulatory drugs (IMiDs). Here we report that glutamine synthetase (GS) is an endogenous substrate of CRL4(CRBN). Upon exposing cells to high glutamine concentration, GS is acetylated at lysines 11 and 14, yielding a degron that is necessary and sufficient for binding and ubiquitylation by CRL4(CRBN) and degradation by the proteasome. Binding of acetylated degron peptides to CRBN depends on an intact thalidomide-binding pocket but is not competitive with IMiDs. These findings reveal a feedback loop involving CRL4(CRBN) that adjusts GS protein levels in response to glutamine and uncover a new function for lysine acetylation.

Description of Mycolicibacterium arenosum sp. nov. Isolated from Coastal Sand on the Yellow Sea Coast.

A Gram-staining-positive, aerobic, non-spore-forming bacterium was isolated from coastal sand samples from Incheon in the Republic of Korea and designated as strain CAU 1645T. The optimum conditions for growth were observed at 30 °C in growth media containing 1% (w/v) NaCl at pH 9.0. The predominant respiratory quinone was MK-9 and the major fatty acids were C16:0, C17:1 w7c, and summed feature 7. Similarly, the 16S rRNA gene sequence exhibited the highest similarity with Mycolicibacterium bacteremicum DSM 45578T and Mycolicibacterium neoaurum JCM 6365T, both of which exhibited similarity rates of 97.2%. The genomic DNA G+C content was 68.2%. The whole genome of strain CAU 1645T was obtained and annotated with annotation using RAST server. The pan-genome analysis was determined using Prokka, Roary, and Phandango. In the pan-genome analysis, the strain CAU 1645T shared 40 core genes with closely related Mycolicibacterium species, including the AcpM gene, the meromycolate extension acyl carrier protein involved in forming impermeable cell walls in mycobacteria. Therefore, our findings demonstrated that the isolate represents a novel species of the genus Mycolicibacterium, for which we propose the name Mycolicibacterium arenosum sp. nov. The type strain is CAU 1645T (= KCTC 49724T = MCCC 1K07087T).

Development of Certified Reference Material for Amino Acids in Dried Blood Spots and Accuracy Assessment of Disc Sampling.

To achieve the measurement reliability of amino acids used as diagnostic markers in clinical fields, establishing a reference measurement system is required, in which certified reference materials (CRMs) are an essential step in the hierarchy of measurement traceability. This study describes the development of dried blood spot (DBS) CRMs for amino acid analysis with complete measurement traceability to the International System of Units (SI). Six essential amino acids─proline, valine, isoleucine, leucine, phenylalanine, and tyrosine─were analyzed using isotope-dilution liquid chromatography-mass spectrometry (ID-MS). For minimizing measurement bias and uncertainty overestimation, whole spots with 50 µL of whole blood were adopted in the certification. The between-spot homogeneities by whole spot sampling were lower than 2.1%. The relative expanded uncertainties of the six amino acids in the developed DBS CRMs were lower than 5.7% at 95% confidence. The certified values are traceable to SI through both gravimetric preparation and the primary method in certification, ID-MS. Comparison among DBS testing laboratories revealed discrepancies between the whole spot and disc sampling methods. The actual sampling volume was accurately estimated by weighing, which revealed the possibility of underestimation in routine DBS testing. The candidate CRMs can support the standardization of DBS testing for amino acids through the qualification and validation of many kinds of measurement procedures to compensate the measurement bias caused by matrix-specific sampling error.

Muricauda lutisoli sp. nov., isolated from mudflat.

A Gram-negative, nonmotile, rod-shaped bacterium, designated strain CAU 1631T, was isolated from a mudflat sample in the Republic of Korea. Strain CAU 1631T grew optimally at 30 °C, pH 6.5, and 1% (w/v) NaCl solution. Phylogenetic analysis based on 16S rRNA gene sequencing and 92 core genes indicated that strain CAU 1631T is a member of the genus Muricauda and most closely related to Muricauda oceanensis 40DY170T and Muricauda lutimaris SMK-108T (98.1%, both). The draft genome was 3.4 Mb with 3064 protein-coding genes, and the DNA G + C content was 43.3 mol%. The major fatty acids were iso-C15:0, iso-C17:0 3-OH, and iso-C15:0 G, and the major polar lipid was phosphatidylethanolamine. The predominant respiratory quinone was MK-6. Based on the comprehensive taxonomic characterization, strain CAU 1631T is a novel species, for which the name Muricauda lutisoli sp. nov. has been proposed. The type strain is CAU 1631T (= KCTC 82456T = MCCC 1K06088T).

Halomarinibacterium sedimenti gen. nov., sp. nov., a carotenoid pigment-producing bacterium isolated from marine sediment.

A Gram-negative, strictly aerobic, non-motile, rod-shaped bacterial strain CAU 1614T was isolated from a marine sediment sample collected in the Republic of Korea. Optimal growth of strain CAU 1614T proceeded at 30 °C, pH 7.0, and 2% (w/v) NaCl. 16S rRNA gene similarity was lower than 94.5% with genera Aureisphaera, Marinirhabdus, Aureitalea, Gilvibacter, Ulvibacter, and Jejudonia. The highest similarity was with Aureisphaera galaxeae 04OKA003-7T (94.5%). The major cellular fatty acids were iso-C15:0, iso-C16:0, iso-C15:1 G, iso-C16:0 3-OH, and iso-C17:0 3-OH and the predominant menaquinone was MK-6. The polar lipids were phosphatidylethanolamine, phosphoglycolipid, an unidentified lipid, two unidentified aminolipids, and an unidentified glycolipid. The draft genome of strain CAU 1614T was 3.9 Mb and DNA G+C content was 36.0 mol%. On the basis of the phenotypic, chemotaxonomic, and genomic data, strain CAU 1614T presents a novel genus in the family Flavobacteriaceae, for which the name Halomarinibacterium sedimenti gen. nov., sp. nov. is proposed. The type strain is CAU 1614T (= KCTC 82457T = MCCC 1K06083T).

Arenibacterium arenosum sp. nov., isolated from sea sand.

A Gram-negative, non-motile, short rod-shaped aerobic bacterial strain CAU 1593T was isolated from a coastal sand sample collected in the Republic of Korea. Cells of strain CAU 1593T grew optimally at 30 °C and pH 7.5 in 4% (w/v) NaCl. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CAU 1593T had the highest similarity to Arenibacterium halophilum (97.5%). The whole genome of strain CAU 1593T was 3,979,826 bp with 26 contigs, and the DNA G + C content was 64.3 mol%. The major fatty acid of strain CAU 1593 T was summed feature 8 (C18:1 ω7c/C18:1 ω6c). Q-10 was the only respiratory quinone. The polar lipids were composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two phosphoglycolipids, an unidentified glycolipid, and an unidentified lipid. Based on the results of chemotaxonomic, phylogenetic, and phenotypic analyses, strain CAU 1593T represents a novel species in the genus Arenibacterium, which is named Arenibacterium arenosum sp. nov. The type strain is CAU 1593T (= KCTC 82402T = MCCC 1K05671T).

Measurement comparability of insulin assays using conventional immunoassay kits.

BACKGROUND: The standardization of measurement aims to achieve comparability of results regardless of the analytical methods and the laboratory where analyses are carried out. In this paper, a comparison of results from several immunoassay-based insulin analysis kits is described, and the steps necessary to improve comparability are discussed. METHODS: Four manual enzyme-linked immunosorbent assay (ELISA) kits produced by Mercodia, Alpco, Epitope Diagnostics, and Abcam, and three automated chemiluminescent (CLIA) insulin assay kits (Siemens Centaur XP, Unicel Dxl800, Cobas e801) were compared by analyzing human serum samples and certified reference materials for human insulin. RESULTS: The seven evaluated assay kits showed substantial discrepancies in the results, with relative standard deviation ranges between 1.7% and 23.2%. We find that the traceability chains and the unit conversion factors are not yet harmonized, and current reference materials for insulin are not applicable for immunoassay-based method validation due to the use of different matrices. CONCLUSIONS: The findings suggest the need to fine tune insulin analysis methods, measurement traceability, and any conversion factor used in post-analysis steps in accordance with the necessity for standardization.

Halarcobacter arenosus sp. nov., isolated from marine sediment.

A Gram-stain-negative, slightly curved, rod-shaped bacterial strain CAU 1517T was isolated from marine sediment in Busan, the Republic of Korea. The taxonomic position of strain CAU 1517T was investigated via a polyphasic approach comprising phenotypic, chemotaxonomic and phylogenetic properties. Strain CAU 1517T grew optimally at 30 °C, pH 7.5 and in the presence of 7% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequencing revealed that strain CAU 1517T belongs to the genus Halarcobacter and is most closely related to Halarcobacter bivalviorum LMG 26154T (96.5% similarity). The average nucleotide identity and digital DNA-DNA hybridization values between strain CAU 1517T and members of genus Halarcobacter ranged from were 76.7-78.0% and 19.5-21.2%, respectively. The strain contained menaquinone-6 (MK-6) as the only respiratory quinone, and C16:0, summed feature 3 (C16:1 ω7c/C16:1 ω6c), and summed feature 8 (C18:1ω7c/C18:1ω6c) as the major fatty acids. The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylethanolamine, and two unidentified aminophospholipids. The G+C content was 28.2 mol%. Therefore, it has been demonstrated that the isolate represents a novel species of the genus Halarcobacter, for which the name Halarcobacter arenosus sp. nov., is proposed. The type strain is CAU 1517T (=KCTC 72232T =NBRC 113955T).

Candidate reference measurement procedure based on HPAEC-PAD for the development of certified reference materials for monosaccharides in serum.

To achieve the measurement reliability of monosaccharides used as diagnostic markers in clinical fields, it is essential to establish certified reference materials (CRMs). The purpose of this study is to develop a serum CRM by adopting high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) as a new candidate reference measurement procedure for the measurement of glucose and galactose, common diagnostic markers of diabetes and galactosemia, respectively. Using various monosaccharides as internal standards, the accuracy of the HPAEC-PAD method was tested by measuring glucose CRM following treatment with three different deproteinization methods: ultrafiltration, protein precipitation by trichloroacetic acid (TCA), and protein precipitation by acetonitrile. Results showed that ultrafiltration and 5% TCA provided good accuracy with every tested monosaccharide as the internal standard. Accordingly, serum samples in this study were treated by ultrafiltration after adding 2-deoxy-D-glucose and arabinose, which were selected as internal standards for galactose and glucose, respectively. Both intra- and inter-day recovery tests showed good precision and accuracy within 2%. From the serum CRM batches prepared at two levels, 11 units were analyzed by exact-matched calibration methods, and the mass fractions of galactose and glucose were determined via HPAEC-PAD. The between-unit relative standard deviations were not more than 1.5%, showing homogeneity. The expanded uncertainties (%) of galactose and glucose for both levels were less than 3.6% and 2.3% at 95% confidence. The HPAEC-PAD method presented in this study can significantly improve the accuracy and precision of simultaneous monosaccharide analysis, allowing for the development of further serum CRMs for monosaccharides. Graphical abstract.

Stable Isotope Labeled DNA: A New Strategy for the Quantification of Total DNA Using Liquid Chromatography-Mass Spectrometry.

Conventional DNA quantification methods require a DNA purification step that limits their reliability in estimating the original DNA amount, especially in complex matrix. To overcome this limitation, we developed a method to calibrate the variable DNA extraction efficiencies during the purification process, allowing for the accurate quantification of DNA in complex matrix. This method is based on isotope dilution-liquid chromatography-mass spectrometry using stable isotope labeled DNA (SILD) as an internal standard. Steps include spiking prepared SILD into samples, purification, enzymatic hydrolysis, and detection of DNA monomers via mass spectrometry, where the spiked SILD is expected to behave the same as the target DNA throughout the entire procedure. We show that the mean recoveries of four different DNA purification kits were dramatically improved by using the SILD internal standard, both for Escherichia coli and human genomic DNA. As standards for calibration, deoxyribonucleoside monophosphates and purified genomic DNA were tested, with genomic DNA from corresponding species found to calibrate the variable extraction efficiencies more effectively. With this successful calibration, our newly developed procedure enables International System of Units-traceable quantification of total DNA in complex matrix.

Enhancing the accuracy of measurement of small molecule organic biomarkers.

Over two decades, the Organic Analysis Working Group (OAWG) of the Consultative Committee for Amount of Substance: Metrology in Chemistry and Biology (CCQM) has organized a number of comparisons for clinically relevant small molecule organic biomarkers. The aim of the OAWG community is to be part of the coordinated international movement towards accuracy and comparability of clinical measurements that will, in turn, minimize the wastage of repeat testing and unnecessary therapy to create a sustainable healthcare industry. International and regional directives/requirements on metrological traceability of calibrators and control materials are in place. Metrology institutes worldwide maintain infrastructure for the practical realization of metrological traceability and demonstrate the equivalence of their measurement capabilities through participation in key comparisons organized under the auspices of the CCQM. These institutes provide certified reference materials, as well as other dedicated value-assignment services benefiting the in-vitro diagnostic (IVD) industry, reference (calibration) laboratories and the clinical chemistry laboratories. The roles of these services in supporting national, regional, and international activities to ensure the metrological traceability of clinical chemistry measurements are described. Graphical abstract.

The Intake of Taurine and Major Food Source of Taurine in Elementary School Children in Korea.

Taurine is a ß-amino acid found most broadly distributed in human body, abundant in animal foods, and has an antioxidative function. Current nutritional intake and dietary habits of children in elementary schools show low level of the intake of vegetable foods and high level of the intake of processed foods and fast foods; this necessitates the emphasis of the intake of antioxidative nutrients for children. On account of the less consumption of vegetable foods as a main source of antioxidative nutrients for elementary school children, animal foods containing abundant amount of taurine can be preferably taken as an alternative foods therefor. Many previous studies have reported the protein intake of the children in elementary schools so far. However, the studies, reported the intake of taurine of elementary school children, are few. Thus, this study analyzed taurine and nutrients intake for children in Daegu, Korea. The average daily energy intake of the children was 153 ± 155 mg/day. The mean taurine intake values are followed; 27.6 ± 11.6 mg/day in the Q1 group, 61.2 ± 10.0 mg/day in the Q2 group, 137.7 ± 51.1 mg/day in the Q3 group, and 385.9 ± 123.6 mg/day in the Q4 group (p < .001). Q3 and Q4 groups showed significantly higher level of the intake of vitamin D, vitamin B12, Calcium, and folate than those of Q1 and Q2 groups. In the study, foods that affected the intake of taurine were as followed; fish and shellfish (79%), meat (14%), seaweed (5%), and other food products (2%).As a consequence, Taurine intake appears to be affected by seafood intake, and if seafood is consumed primarily, the amount of energy intake would be appropriate and will contribute to the increase of intakes of taurine, calcium and vitamin D.

ß-cell serotonin production is associated with female sex, old age, and diabetes-free condition.

Serotonin is known to be present in pancreatic ß-cells and to play several physiological roles, including insulin secretion, ß-cell proliferation, and paracrine inhibition of α-cells. However, the serotonin production of different cell lines and islets has not been compared based on age, sex, and diabetes related conditions. Here, we directly compared the serotonin concentrations in ßTC and MIN6 cell lines, as well as in islets from mice using ultra-performance liquid chromatography tandem mass spectrometry. The average serotonin concentration was 5-10 ng/mg protein in the islets of male and non-pregnant female mice. The serotonin level was higher in females than males at 8 weeks, although there was no difference at 1 year. Furthermore, we observed serotonin by immunofluorescence staining in the pancreatic tissues of mice and human. Serotonin was detected by immunofluorescence staining in a portion of ß-cells from islets of old female mice, but not of male or young female mice. A similar pattern was observed in human pancreas as well. In humans, serotonin production in ß-cells was associated with a diabetes-free condition. Thus, serotonin production in ß-cells was associated with old age, female sex, and diabetes-free condition.

Quantification of recombinant human erythropoietin by amino acid analysis using isotope dilution liquid chromatography-tandem mass spectrometry.

Herein, we describe an accurate method for protein quantification based on conventional acid hydrolysis and an isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry method. The analyte protein, recombinant human erythropoietin (rhEPO), was effectively hydrolyzed by incubation with 8 mol/L hydrochloric acid at 130 °C for 48 h, in which at least 1 µmol/kg of rhEPO was treated to avoid possible degradation of released amino acids during hydrolysis. Prior to hydrolysis, sample solution was subjected to ultrafiltration to eliminate potential interfering substances. In a reversed-phase column, the analytes (phenylalanine, proline, and valine) were separated within 3 min using gradient elution comprising 20 % (v/v) acetonitrile and 10 mmol/L ammonium acetate, both containing 0.3 % (v/v) trifluoroacetic acid. The optimized hydrolysis and analytical conditions in our study were strictly validated in terms of accuracy and precision, and were suitable for the accurate quantification of rhEPO. Certified rhEPO was analyzed using a conventional biochemical assay kit as an additional working calibrant for the quantification of EPO and improved the accuracy. The optimized protocol is suitable for the accurate quantification of rhEPO and satisfactorily serves as a reference analytical procedure for the certification of rhEPO and similar proteins.

Simultaneous quantification of 11 antiepileptic drugs using limited isotope-labeled internal standards in LC-MS/MS: An accuracy assessment.

This study aims to establish an LC-MS/MS method to simultaneously analyze 11 antiepileptic drugs with a particular focus on maintaining accuracy while reducing the number of isotope-labeled internal standards employed for cost-effectiveness. By applying a water/acetonitrile gradient elution containing 0.1 % formic acid and 2 mM ammonium formate as the mobile phase, optimal sensitivity for the target drugs could be obtained in positive ESI mode in LC-MS/MS. After optimizing various extraction techniques, extraction with 70 % acetonitrile was selected as it provided good recoveries (>93 %) for all targets without matrix effects. Accuracies within 3 % were achieved from the combination of six internal standards, while accuracies of 5 % and 10 % were obtained by reducing the number of internal standards to four and two, respectively, for more economical analysis. The accuracy of the established method was maintained in hyperglycemia, hyperlipidemia, and hyperalbuminemia sera, suggesting that it can be successfully applied to individual serum samples with various properties.

Dried blood spot sampling coupled with liquid chromatography-tandem mass for simultaneous quantitative analysis of multiple cardiovascular drugs.

Dried Blood Spots (DBS) revolutionize therapeutic drug monitoring using LC-MS for the precise quantification of cardiovascular drugs (CDs), enabling personalized treatment adapted to patient-specific pharmacokinetics with minimal invasiveness. This study aims to achieve simultaneous quantification of eight CDs in DBS, overcoming physicochemical challenges. A two-step protein precipitation method was used for simple and precise sample preparation. The drugs were analyzed using LC-MS/MS in ESI positive-ion mode, showing high sensitivity and linearity, with a correlation coefficient (r2) exceeding 0.999, after being separated on a reversed-phase chromatography by gradient elution of DW-acetonitrile containing 0.1 % formic acid + 2 mM ammonium formate. The validation results indicate good selectivity, with no observed matrix effect and carry-over. The intra- and inter-day accuracy and precision were within 6 % for most drugs, except for digoxin and deslanoside at low therapeutic levels where the variation was within 20 %. Stability tests confirmed suitable DBS handling and storage conditions, indicating drug stability for at least 30 days at room temperature. The analysis of whole spot has demonstrated remarkable precision and reliability in all target drugs. The analysis of 3 mm internal diameter discs, punched in and out of DBS, presumed to contain 3 µL of blood, showed acceptable accuracy for most drugs, with less polar drugs like digoxin and deslanoside showing lower accuracy, indicating a need for further correction due to non-uniform drug distribution. Consequently, the developed LC-MS/MS method enables the quantification of multiple CDs in a single DBS analysis, while suggesting the potential for accuracy-based analysis.

The reprotoxic adverse side effects of neurogenic and neuroprotective drugs: current use of human organoid modeling as a potential alternative to preclinical models.

The management of neurological disorders heavily relies on neurotherapeutic drugs, but notable concerns exist regarding their possible negative effects on reproductive health. Traditional preclinical models often fail to accurately predict reprotoxicity, highlighting the need for more physiologically relevant systems. Organoid models represent a promising approach for concurrently studying neurotoxicity and reprotoxicity, providing insights into the complex interplay between neurotherapeutic drugs and reproductive systems. Herein, we have examined the molecular mechanisms underlying neurotherapeutic drug-induced reprotoxicity and discussed experimental findings from case studies. Additionally, we explore the utility of organoid models in elucidating the reproductive complications of neurodrug exposure. Have discussed the principles of organoid models, highlighting their ability to recapitulate neurodevelopmental processes and simulate drug-induced toxicity in a controlled environment. Challenges and future perspectives in the field have been addressed with a focus on advancing organoid technologies to improve reprotoxicity assessment and enhance drug safety screening. This review underscores the importance of organoid models in unraveling the complex relationship between neurotherapeutic drugs and reproductive health.

Melatonin and resveratrol alleviate molecular and metabolic toxicity induced by Bisphenol A in endometrial organoids.

Bisphenol A (BPA), a widespread environmental contaminant, poses concerns due to its disruptive effects on physiological functions of the uterine endometrium. In contrast, melatonin (MT) and Resveratrol (RSV) are under scrutiny for their potential protective roles against BPA-induced damage. For the efficacy and ethical concerns in the animal test, endometrial organoids, three-dimensional models mimicking endometrium, serve as crucial tools for unraveling the impact of environmental factors on reproductive health. This study aimed to comprehensively characterize the morphological, molecular and metabolic responses of porcine endometrial organoids to BPA and assess the potential protective effects of MT and RSV. Porcine uteri were prepared, digested with collagenase, mixed with Matrigel, and incubated at 38°C with 5 % CO2. Passaging involved dissociation through trypsin-EDTA treatment and subculturing. The culture medium was refreshed every 2-3 days. To investigate the environmental impact on reproductive health, endometrial organoids were treated with BPA (0.5 µM), MT (with/without BPA at 0.1 µM), and/or RSV (10 µM). Various molecular screening using gene expression, western blotting, immunofluorescence staining, and metabolites profiling were assessed the effects of BPA, MT, and RSV in terms of cell viability, morphology, reproductivity, and metabolism alteration in the endometrial organoids. As expected, BPA induced structural and molecular disruptions in organoids, affecting cytoskeletal proteins, Wnt/ß-catenin signaling, and epithelial/mesenchymal markers. It triggered oxidative stress and apoptotic pathways, altered miRNA expression, and disrupted the endocannabinoid system. The level of glucose, galactose, and essential amino acids were increased or decreased by approximately 1.5-3 times in BPA-treated groups compared to the control groups (p-value < 0.05), indicating metabolic changes. Moreover, MT and RSV treated groups exhibited protective effects, mitigating BPA-induced disruptions across multiple pathways. For the first time, our study models endometrial organoids, advancing understanding of environmental impacts on reproductive health.

The secreted protein Amuc_1409 from Akkermansia muciniphila improves gut health through intestinal stem cell regulation.

Akkermansia muciniphila has received great attention because of its beneficial roles in gut health by regulating gut immunity, promoting intestinal epithelial development, and improving barrier integrity. However, A. muciniphila-derived functional molecules regulating gut health are not well understood. Microbiome-secreted proteins act as key arbitrators of host-microbiome crosstalk through interactions with host cells in the gut and are important for understanding host-microbiome relationships. Herein, we report the biological function of Amuc_1409, a previously uncharacterised A. muciniphila-secreted protein. Amuc_1409 increased intestinal stem cell (ISC) proliferation and regeneration in ex vivo intestinal organoids and in vivo models of radiation- or chemotherapeutic drug-induced intestinal injury and natural aging with male mice. Mechanistically, Amuc_1409 promoted E-cadherin/ß-catenin complex dissociation via interaction with E-cadherin, resulting in the activation of Wnt/ß-catenin signaling. Our results demonstrate that Amuc_1409 plays a crucial role in intestinal homeostasis by regulating ISC activity in an E-cadherin-dependent manner and is a promising biomolecule for improving and maintaining gut health.