Epithelial Structures Research Articles

Unveiling the Etiopathogenic Role of Epstein-Barr Virus in Periodontitis.

Periodontitis, a prevalent and costly oral disease, remains incompletely understood in its etiopathogenesis. The conventional model attributes it to pathogenic bacteria, but emerging evidence suggests dysbiosis involving bacteria, herpesviruses, and an exaggerated host immune response. Among herpesviruses, Epstein-Barr virus (EBV) closely links to severe periodontitis, yet the mechanisms underlying EBV-related pathogenesis remain elusive. This study examined the presence, methylation patterns, and infection states of EBV in gingival tissues from healthy patients and those with periodontitis. It also assessed gene expression differences associated with EBV through whole-genome transcriptomic profiling in healthy and periodontitis-affected tissues. EBV DNA was found at similar frequencies in healthy and periodontitis tissues, suggesting common EBV infection even before disease manifestation. In healthy tissues, mostly unmethylated EBV genomes indicated lytic infection in gums, consistent with the literature on lytic EBV spread in epithelia and continual significant virus release in the saliva of healthy carriers. Conversely, EBV DNA in periodontitis tissues showed both methylated and unmethylated patterns, suggesting a mix of latent and lytic genomes. This indicates the coexistence of latent EBV in B-cells and lytic EBV in plasma cells (PCs), linking EBV presence with both cell types in periodontitis. Whole-genome transcriptomic analysis revealed distinct expression profiles in EBV-positive periodontitis tissues, with upregulated genes associated with inflammatory/immune responses and B-cell and PC markers, while downregulated genes were related to epithelial structure and organization. The EBV-positive periodontitis signature differed distinctly from that of EBV-positive healthy gums, eliciting only a typical viral-induced immune response. These findings provide new insights into EBV physiopathology in the gum, notably assigning a direct etiopathogenetic contribution to EBV in periodontitis. The results suggest a model where EBV can commonly, and apparently asymptomatically, spread in healthy gingiva but may also aggravate inflammation in the context of gum dysbiosis, involving infiltration of B-cells and PCs and loss of epithelial integrity.

Therapeutic Potential of Cajanus cajan (L.) Millsp. Leaf Extract in Modulating Gut Microbiota and Immune Response for the Treatment of Inflammatory Bowel Disease.

Background/Objectives: Inflammatory bowel disease (IBD) is a persistent inflammatory condition affecting the gastrointestinal tract, distinguished by the impairment of the intestinal epithelial barrier, dysregulation of the gut microbiota, and abnormal immune responses. Cajanus cajan (L.) Millsp., traditionally used in Chinese herbal medicine for gastrointestinal issues such as bleeding and dysentery, has garnered attention for its potential therapeutic benefits. However, its effects on IBD remain largely unexplored. Methods: In this study, the major compounds from Cajanus cajan leaf extract (CCLE) were initially characterized by LCMS-IT-TOF. The IBD model was developed in C57BL/6 mice by administering continuous 4% (w/v) dextran sodium sulfate (DSS) aqueous solution over a period of seven days. The body weight, colon length, disease activity index (DAI), and histopathological examination using hematoxylin and eosin (H&E) staining were performed in the IBD model. The levels of the main inflammatory factors, specifically TNF-α, IL-1β, IL-6, and myeloperoxidase (MPO), were quantified by employing enzyme-linked immunosorbent assay (ELISA) kits. Additionally, the levels of tight junction proteins (ZO-1, Occludin) and oxidative stress enzymes (iNOS, SOD1, CAT) were investigated by qPCR. Subsequently, flow cytometry was employed to analyze the populations of various immune cells within the spleen, thereby assessing the impact of the CCLE on the systemic immune homeostasis of IBD mice. Finally, 16S rDNA sequencing was conducted to examine the composition and relative abundance of gut microbiota across different experimental groups. In addition, molecular docking analysis was performed to assess the interaction between the principal components of CCLE and the aryl hydrocarbon receptor (AHR). Results: We identified seven bioactive compounds in CCLE: catechin, cajachalcone, 2-hydroxy-4-methoxy-6-(2-phenylcinyl)-benzoic acid, longistylin A, longistylin C, pinostrobin, amorfrutin A, and cajaninstilbene acid. Our results demonstrated that oral administration of CCLE significantly alleviates gastrointestinal symptoms in DSS-induced IBD mice by modulating the balance of gut-derived pro- and anti-inflammatory cytokines. This modulation is associated with a functional correction in M1/M2 macrophage polarization and the Th17/Treg cell balance in splenic immune cells, as well as shifts in the populations of harmful bacteria (Erysipelatoclostridium and Staphylococcus) and beneficial bacteria (Odoribacter, unidentified Oscillospiraceae, Lachnoclostridium, and Oscillibacter) in the gut. Furthermore, cajaninstilbene acid, longistylin A, and longistylin C were identified as potential AhR agonists. Conclusions: The present results suggested that CCLE, comprising stilbenes like cajaninstilbene acid, longistylin A, and longistylin C, protects the epithelial barrier's structure and function against DSS-induced acute IBD by restoring gut microbiota balance and systemic immune response as AhR agonists. Overall, CCLE represents a promising natural product-based therapeutic strategy for treating IBD by restoring gut microbiota balance and modulating systemic immune responses.

A spatiotemporal and machine-learning platform facilitates the manufacturing of hPSC-derived esophageal mucosa.

Human pluripotent stem cell-derived tissue engineering offers great promise for designer cell-based personalized therapeutics, but harnessing such potential requires a deeper understanding of tissue-level interactions. We previously developed a cell replacement manufacturing method for ectoderm-derived skin epithelium. However, it remains challenging to manufacture the endoderm-derived esophageal epithelium despite possessing a similar stratified epithelial structure. Here, we employ single-cell and spatial technologies to generate a spatiotemporal multi-omics cell census for human esophageal development. We identify the cellular diversity, dynamics, and signal communications for the developing esophageal epithelium and stroma. Using Manatee, a machine-learning algorithm, we prioritize the combinations of candidate human developmental signals for invitro derivation of esophageal basal cells. Functional validation of Manatee predictions leads to a clinically compatible system for manufacturing human esophageal mucosa.

Low molecular weight galactomannan alleviates diarrhea induced by senna leaf in mice via intestinal barrier improvement and gut microbiota modulation.

Low molecular weight galactomannan (LMGM), a soluble dietary fibre derived from guar gum, is recognized for its prebiotic functions, including promoting the growth of beneficial intestinal bacteria and the production of short-chain fatty acids, but the mechanism of alleviating diarrhea is not fully understood. This study established an acute diarrhea mouse model using senna leaf decoction and evaluated the therapeutic effects of LMGM by monitoring diarrhea scores, loose stool prevalence, intestinal tissue pathology and gene expression, and gut microbiota composition and metabolisms. The results indicated that LMGM significantly reduced diarrhea scores and loose stool prevalence within two hours post-treatment. Hematoxylin and eosin staining and quantitative real-time polymerase chain reaction analysis revealed that LMGM improved intestinal epithelial structure and up-regulated the expression of zonula occludens 1, occludin, mucin 2, aquaporin 3, and aquaporin 4 in ileum, jejunum, and colon tissues. Moreover, LMGM increased the abundance of beneficial bacteria such as Lactobacillaceae and Lachnospiraceae, and decreased Prevotellaceae in the cecum. Furthermore, LMGM promoted short-chain fatty acid production and reduced ammonia nitrogen and skatole concentrations in the intestinal content. The study suggests that LMGM could serve as a functional prebiotic for diarrhea alleviation, potentially by enhancing the intestinal barrier, modulating water transportation, and regulating the microbiota composition.

ULTRASTRUCTURAL CHANGES IN THE KIDNEYS OF RATS DURING THE EARLY AND LATE STAGES OF ACUTE RESPIRATORY DISTRESS SYNDROME

Introduction. Acute respiratory distress syndrome (ARDS) is characterized by severe hypoxemia and activation of the inflammatory cascade, which leads to multi-organ failure. Kidneys are one of the organs commonly affected by ARDS. However, the extent of ultrastructural changes in the parenchyma and stroma of the kidneys at different stages of ARDS progression remains unclear. The aim of this study was to use transmission electron microscopy to study the main components of the kidneys in rats at the early and late stages of experimentally induced ARDS. Materials and methods. The study included 24 healthy, sexually mature male rats weighing 200–220 g, divided into three groups: intact animals, 24 hours post-ARDS induction, and 14 days after the start of the experiment. ARDS was modeled by intranasal administration of lipopolysaccharide (5 mg/kg body weight) under anesthesia with ketamine at a dose of 50 mg/kg. Terminal anesthesia was performed using sodium thiopental at a dose of 150 mg/kg. Results: Submicroscopic examination of the kidneys from intact rats revealed no species-specific structural abnormalities. In the early stage of ARDS (24 hours post-modeling), renal corpuscles showed signs of vascular endothelial damage, fragmentation of the glomerular basement membrane, and degeneration of organelles and nuclear components in tubular epithelial cells. By day 14 of ARDS progression, signs of damage to the filtration barrier persisted, resulting in podocyte swelling. However, partial restoration of the tubular epithelial cell structure was observed, indicating the onset of the resolution phase of ARDS, as evidenced by the gradual recovery of the kidney ultrastructure. Conclusions. During the early stage of modeled ARDS, kidney damage manifests through destructive changes in the glomerular apparatus and renal tubular epithelial cells. In the late stage, although glomerular damage remains, partial restoration of the renal tubular epithelial cell structure is still evident.

Macro- and microscopic anatomy of the digestive tract in the red-eared slider (Emydidae: Trachemys scripta elegans).

The red-eared sliders (Emydidae: Trachemys scripta) is characterised by a high adaptability to a variety of environment and threatens the habitat of Japanese native species. The ability to digest a variety of diets may attribute to the high adaptive capacity of this species to various environments, however, the digestive morphology remains scarcely described in red-eared sliders. In this study, we investigated the macro- and microscopic anatomy of the esophagus, stomach, small intestine, and large intestine in red-eared sliders. All segments of the digestive tract had longitudinal mucosal folds, the height and width of which varied in each segment of the digestive tract. The stomach had the highest and widest mucosal folds. The mucosal folds in the proximal-to-middle small intestine exhibited a zigzag shape, whereas those in the distal small intestine were linear. The wall of the digestive tract regularly consisted of mucosa, submucosa, tunica muscularis, and tunica adventitia or serosa. In each segment of the digestive tract, the epithelial structure was different. The esophagus and small intestine were lined by the pseudostratified columnar epithelium. In both segments, the basal part of the pseudostratified epithelium included proliferating cell nuclear antigen (PCNA)-positive proliferating cells. The stomach and large intestine were lined by the simple columnar epithelium. In the stomach and large intestine, PCNA-positive proliferating cells were present in the neck of the proper gastric gland and crypt-like structures, respectively. The proper gastric gland was composed of oxynticopeptic and mucous cells. This study revealed the detailed macro- and microscopic anatomy of the digestive tract in red-eared sliders. Overall, our findings may provide an anatomical basis for understanding the relationship between morphology and function in the digestive tract of turtles.

Convergent Evolution Has Led to the Loss of Claw Proteins in Snakes and Worm Lizards.

The evolution of cornified skin appendages, such as hair, feathers, and claws, is closely linked to the evolution of proteins that establish the unique mechanical stability of these epithelial structures. We hypothesized that the evolution of the limbless body anatomy of the Florida worm lizard (Rhineura floridana) and the concomitant loss of claws had led to the degeneration of genes with claw-associated functions. To test this hypothesis, we investigated the evolution of three gene families implicated in epithelial cell architecture, namely type I keratins, type II keratins, and genes of the epidermal differentiation complex in R. floridana in comparison with other squamates. We report that the orthologs of mammalian hair and nail keratins have undergone pseudogenization in R. floridana. Likewise, the epidermal differentiation complex genes tentatively named EDYM1 and EDCCs have been lost in R. floridana. The aforementioned genes are conserved in various lizards with claws, but not in snakes. Proteomic analysis of the cornified claws of the bearded dragon (Pogona vitticeps) confirmed that type I and type II hair keratin homologs, EDYM1 and EDCCs, are protein components of claws in squamates. We conclude that the convergent evolution of a limbless body was associated with the convergent loss of claw keratins and differentiation genes in squamates.

Establishment of a chronic biliary disease mouse model with cholecystoduodenal anastomosis for intestinal microbiome preservation.

Chronic biliary disease, including cholangitis and cholecystitis, is attributed to ascending infection by intestinal bacteria. Development of a mouse model for bile duct inflammation is imperative for the advancement of novel therapeutic approaches. Current models fail to replicate the harmful bacterial influx to the biliary tract observed in humans and spread of inflammation to the liver. Therefore, we aimed to establish an animal model of biliary disease that faithfully replicates the mechanisms of human diseases. To establish a cholecystoduodenal anastomosis model capable of mimicking the mechanisms of ascending infection and inflammation observed in human biliary diseases. We established a mouse biliary disease model by directly connecting the gallbladder and duodenum, enabling ascending infection into the biliary tract without traversing the sphincter of Oddi. In the cholecystoduodenal anastomosis mouse model, we observed impaired epithelial structure, wall thickening, and macrophage recruitment in the gallbladder. Despite the absence of postoperative antibiotics, we detected no changes in serum proinflammatory cytokine levels, indicating no systemic inflammation. Moreover, patency between the gallbladder and duodenum was confirmed via common bile duct ligation. Injection of patient-derived pathogenic bacteria into bile duct-ligated mice led to ascending infection, which significantly increased proinflammatory cytokine mRNA expression in the liver, duodenum, and ileum. These results indicate that our mouse model exhibited a direct connection between the gallbladder and duodenum, leading to ascending infection and closely mimicking the clinical features of biliary diseases observed in humans. The cholecystoduodenal anastomosis mouse model is an effective chronic biliary disease model with significant relevance in the development of microbiome-based therapies for the prevention and treatment of biliary disease.

The development of orofacial complex in bats: Implications for orofacial clefting.

Orofacial morphology in mammals plays a critical role in essential life functions such as feeding and communication, which are influenced by the shapes of these anatomical structures. Bats are known to exhibit highly diversified orofacial morphotypes within their clade, reflecting their varied diets and echolocation behaviors. The presence of bony discontinuities between the premaxilla and maxilla or among the premaxillae is a notable feature of bat orofacial morphology, observed in certain lineages. It is suggested that these unique orofacial morphotypes, not generally found in other mammals, have evolved in relation to dietary adaptations rather than merely for echolocation mode. Until now, the developmental background of the bony discontinuities in the bat orofacial complex has been insufficiently investigated. Here, we present a comparative study of the chondrocranium and epithelial organs in the orofacial complex of three bat species: Cynopterus sphinx, Rhinolophus malayanus, and Vespertilio sinensis. Our observations indicate that the preceding morphogenesis of orofacial cartilage and epithelial structures is remarkably different among these three species. In C. sphinx and V. sinensis, the region forming from the regression of the palatine process of the premaxilla was filled with orofacial cartilage and epithelial structures. We also found that the clefted morphology observed in R. malayanus and V. sinensis was formed via contrastingly divergent developmental processes. Midline clefts among Yangochiroptera have been previously categorized to represent a uniform morphotype, but our study highlights that attributing midline clefts into a singular category should be revisited, advocating for a nuanced categorization of cleft morphology based on their morphogenetic patterns. Further research on the bat orofacial complex may enhance our understanding of bat evolutionary diversification and offer insights into the developmental mechanisms of human cleft palate.

Spatial transcriptomics and in situ immune cell profiling of the host ectocervical landscape of HIV infected Kenyan sex working women.

Chronic immune activation is a hallmark of human immunodeficiency virus (HIV) infection that significantly impacts disease pathogenesis. However, in-depth studies characterizing the immunological landscape of the ectocervix during chronic HIV infection remain scarce despite the importance of this tissue site for HIV transmission. Ectocervical tissue samples were obtained from antiretroviral-naïve HIV-seropositive and -seronegative Kenyan female sex workers. These samples were assessed by spatial transcriptomics and Gene Set Enrichment Analysis. We further performed multi-epitope ligand cartography (MELC) using an in situ staining panel that included 17 markers of primarily T cell-mediated immune responses. Spatial transcriptomics revealed tissue-wide immune activation encompassing immune responses associated with chronic HIV infection. First, both the epithelial and submucosal compartments showed diverse but significant upregulation of humoral immune responses, as indicated by the expression of several antibody-related genes. Second, an antiviral state-associated cellular immunity was also observed in the HIV-seropositive group, characterized by upregulation of genes involved in interferon signaling across the mucosal tissue and a more spatially restricted mucosal expression of genes related to T cell activity and effector functions relative to the HIV-seronegative group. Additionally, HIV associated structural alterations were evident within both compartments. Downregulated genes across the epithelium were mainly linked to epithelial integrity, with the outer layer involved in terminal differentiation and the inner layer associated with epithelial structure. MELC analysis further revealed a significantly increased ectocervical leukocyte population in HIV-seropositive participants, primarily driven by an increase in CD8+ T cells while the CD4+ T cell population remained stable. Consistent with our spatial transcriptomics data, T cells from HIV-seropositive participants showed an increased effector phenotype, defined by elevated expression of various granzymes. By combining spatial transcriptomics and MELC, we identified significant HIV-associated cervical immune activity driven by induction of both T and B cell activity, together with a general antiviral state characterized by sustained interferon induction. These findings underscore that chronic HIV infection is associated with an altered ectocervical mucosal immune landscape years after primary infection. This sheds light on HIV pathogenesis at distant local sites and complements current knowledge on HIV-associated systemic immune activation.

Sustained immune activation and impaired epithelial barrier integrity in the ectocervix of women with chronic HIV infection.

Chronic systemic immune activation significantly influences human immunodeficiency virus (HIV) disease progression. Despite evidence of a pro-inflammatory environment in the genital tract of HIV-infected women, comprehensive investigations into cervical tissue from this region remain limited. Similarly, the consequences of chronic HIV infection on the integrity of the female genital epithelium are poorly understood, despite its importance in HIV transmission and replication. Ectocervical biopsies were obtained from HIV-seropositive (n = 14) and HIV-seronegative (n = 47) female Kenyan sex workers. RNA sequencing and bioimage analysis of epithelial junction proteins (E-cadherin, desmoglein-1, claudin-1, and zonula occludens-1) were conducted, along with CD4 staining. RNA sequencing revealed upregulation of immunoregulatory genes in HIV-seropositive women, primarily associated with heightened T cell activity and interferon signaling, which further correlated with plasma viral load. Transcription factor analysis confirmed the upregulation of pro-inflammatory transcription factors, such as RELA, NFKB1, and IKZF3, which facilitates HIV persistence in T cells. Conversely, genes and pathways associated with epithelial barrier function and structure were downregulated in the context of HIV. Digital bioimage analysis corroborated these findings, revealing significant disruption of various epithelial junction proteins in ectocervical tissues of the HIV-seropositive women. Thus, chronic HIV infection associated with ectocervical inflammation, characterized by induced T cell responses and interferon signaling, coupled with epithelial disruption. These alterations may influence HIV transmission and heighten susceptibility to other sexually transmitted infections. These findings prompt exploration of therapeutic interventions to address HIV-related complications and mitigate the risk of sexually transmitted infection transmission.

The efficacy and active compounds of Chaihuang Qingyi Huoxue granule to Ameliorate intestinal mucosal barrier injury in rats with severe acute pancreatitis by suppressing the HMGB1/TLR4/NF-κB signaling pathway

Intestinal mucosal barrier injury represents a critical complication of severe acute pancreatitis (SAP) without effective treatment. This study investigated the efficacy, underlying mechanism, and responsible active compounds of the traditional Chinese medicinal prescription Chaihuang Qingyi Huoxue granule (CHQY) in treating SAP-induced intestinal mucosal barrier injury. SAP was established in Sprague-Dawley rats via intra-pancreaticobiliary duct infusion of sodium taurocholate, followed by oral CHQY administration (3.15 g/kg every 6 h for 12 and 24 h). Blood and tissues were harvested to assess the severity of pancreatitis, intestinal mucosal barrier integrity, and extent of inflammatory injury. Intestine-absorbing compounds were identified using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS). Our results showed that CHQY treatment effectively mitigated SAP-induced intestinal mucosal injury, as evidenced by improved intestinal epithelial structure, decreased serum levels of intestinal injury markers (d-lactic acid, diamine oxidase, I-FABP, and Zonulin), restored expression of the tight junction protein ZO-1, and reduced serum endotoxin levels. Furthermore, CHQY administration suppressed the expression of proinflammatory mediator HMGB1, its receptor TLR4, and downstream NF-κB signaling in the intestine, leading to downregulated intestinal IL-1β expression and reduced circulating TNF-α and IL-6. UHPLC-HRMS analysis identified 15 intestine-absorbing compounds in CHQY, of which paeoniflorin sulfite and chrysin-7-O-glucuronide independently inhibited TNF-α-induced tight junction loss in IEC-6 cells and mitigated intestinal mucosal barrier injury in SAP rats through suppressing NF-κB signaling. In summary, CHQY ameliorates SAP-induced intestinal mucosal barrier injury by downregulating the proinflammatory HMGB1/TLR4/NF-κB signaling, with efficacy partially attributed to its active compounds paeoniflorin sulfite and chrysin-7-O-glucuronide.

Evaluation of mucosal barrier disruption due to Staphylococcus lugdunensis and Staphylococcus epidermidis exoproteins in patients with chronic rhinosinusitis.

Chronic rhinosinusitis (CRS) is a persistent inflammatory condition of the sinus mucosa. While Staphylococcus aureus has been shown to play a significant role in mucosal barrier disruption in CRS patients, coagulase-negative staphylococci (CoNS) such as Staphylococcus epidermidis and Staphylococcus lugdunensis are also implicated in CRS pathophysiology. This study investigates the effects of exoproteins secreted by planktonic and biofilm forms of clinical isolates of S. epidermidis and S. lugdunensis on the nasal epithelial barrier. Thirty-one clinical isolates of CoNS were grown in planktonic and biofilm forms, and their exoproteins were concentrated. The epithelial barrier structure was assessed by measuring transepithelial electrical resistance (TEER) and the permeability of fluorescein isothiocyanate-dextran. Toxicity and inflammatory response were also studied. Our findings demonstrate that exoproteins from all planktonic forms of S. lugdunensis disrupted the mucosal barrier, whereas only nine of 16 biofilm-derived exoproteins had similar effects. Conversely, 11 of 15 exoproteins from planktonic S. epidermidis significantly disrupted barrier integrity; however, biofilm exoproteins did not. The study also showed that some exoproteins from planktonic S. epidermidis significantly reduced cell viability, while exoproteins from planktonic and biofilm forms of S. lugdunensis and biofilm S. epidermidis did not induce any statistically significant change in cell viability. Notably, four of 16 biofilm exoproteins from S. lugdunensis induced higher interleukin-6 (IL-6) secretion, whereas none of the S. epidermidis isolates showed a significant increase in IL-6 secretion. Our results suggest that CoNS exoproteins may contribute to CRS etiopathogenesis.

4D pathology: translating dynamic epithelial tubulogenesis to prostate cancer pathology.

The Gleason score is the gold standard for grading of prostate cancer (PCa) and is assessed by assigning specific grades to different microscopical growth patterns. Aside from the Gleason grades, individual growth patterns such as cribriform architecture were recently shown to have independent prognostic value for disease outcome. PCa grading is performed on static tissue samples collected at one point in time, whereas in vivo epithelial tumour structures are dynamically invading, branching and expanding into the surrounding stroma. Due to the lack of models that are able to track human PCa microscopical developments over time, our understanding of underlying tissue dynamics is sparse. We postulate that human PCa expansion utilizes embryonic and developmental tubulogenetic pathways. The aim of this study is to provide a comprehensive overview of developmental pathways of normal epithelial tubule formation, elongation, and branching, and relate those to the static microscopical PCa growth patterns observed in daily clinical practise. This study could provide a rationale for the discerned pathological interobserver variability and the clinical outcome differences between PCa growth patterns.

Sorted stem/progenitor epithelial cells of pubertal bovine mammary gland present limited potential to reconstitute an organised mammary epithelium after transplantation.

The development and maintenance of mammary gland tissue depend on the proliferation and differentiation of mammary stem and progenitor cells. Here, we investigated populations of mammary epithelial cells that are potential candidates for bovine mammary gland development using xenotransplantation into mice cleared mammary fat pad. Transplanted mammary explants from 17-month-old Holstein heifers developed outgrowths exhibiting the archetypal morphology and molecular marker distributions of the bovine gland. Xenotransplantation of sorted mammary epithelial cells (CD49fpos) into bovinised fat pads using inactivated bovine fibroblasts resulted in outgrowth developments with 50% take rate, but these lacked the ductal or alveolar epithelial structures of the normal mammary gland. Similar results were obtained with xenografts of candidate bovine mammary epithelial stem cells (CD49fhighCD24pos) or epithelial cells of the basal lineage (CD49fhighCD24neg) which also developed as clumps of cells surrounded by stromal stretches within the mouse adipose tissue. In conclusion, sorted cells showed compromised regenerative potential for epithelial morphogenesis. Further work is therefore needed to identify mammary stem/progenitor cells with full regenerative capabilities for biogenesis of normal mammary gland structure, with milk-secreting function.

A New Diagnosis of Intestinal Metaplasia in Gastric Mucosa: Convolutional Neural Network Based on Visual Auxiliary Method

Abstract Introduction/Objective Intestinal metaplasia (IM) is a common lesion observed in chronic gastric disease and is thought to be associated with the development of gastric cancer. One of the characteristics of IM is the presence of goblet cells. However, differentiating these inconspicuous goblet cells from glandular cavities and gastric mucus cells using traditional algorithms is inefficient. This paper proposes an interpretable auxiliary diagnosis method for gastric mucosal intestinal metaplasia based on a convolutional neural network(CNN). Methods/Case Report The overall flow chart of the algorithm is shown in Figure 1A. The self-constructed gastric mucosa dataset is utilized in this paper for IM. To improve performance, some modifications are applied to the U-Net. The auxiliary diagnosis process incorporates R34W and R34U for segmenting the glands and IM regions. The R34W represents a dual-resolution input network structure (Figure 1B). For the initial stage of the auxiliary diagnosis, the convolutional neural network calculates patches with both large and small fields for gland region segmentation. Then, the IM glands segmentation network is used to classify regions with and without IM lesions. Results (if a Case Study enter NA) The result of the WSI level mask generated after post-processing is shown in Figure 2A. R34W Net offers significant advantages in differentiating open epithelial structures and proper glands. This network effectively differentiates between proper glands and epithelial regions while maintaining precise segmentation, leveraging high-resolution features in a small visual field. The quantitative indexes of gland segmentation networks are shown in Figure 2B. As part of auxiliary diagnosis, the classification of proper glands with intestinal metaplasia achieves the highest accuracy of 0.95 and AUC of 0.975 in the test datasets (Figure 2C). The area proportion of IM regions generated by CNN is shown in Figure 2D. Conclusion This paper provides a visual auxiliary method based on a CNN for the pathological diagnosis of intestinal metaplasia. The goal is to enhance the speed and accuracy of pathologists in performing this diagnosis. Additionally, we propose a CNN with a dual-resolution input structure, which is utilized for segmenting gastric proper glands.

Metformin modulates the TXNIP-NLRP3-GSDMD pathway to improve diabetic bladder dysfunction

To validate the therapeutic efficacy of metformin on diabetic bladder dysfunction (DBD) and further elucidate whether the TXNIP-NLRP3-GSDMD axis serves as a target for metformin in ameliorating DBD. C57BL/6J mice were induced with diet-induced obesity by being fed a high-fat diet (HFD) for 16 weeks. After establishing the model, the mice were treated with metformin for 4 weeks, and their glucose metabolism-related parameters were assessed. Urine spot assays and urodynamic measurements were conducted to reflect the bladder function and urinary behavior in mice, while histological examination was performed to observe morphological changes. Western blot analysis was employed to measure the expression levels of pyroptotic factors such as TXNIP, NLRP3, GSDMD, and tight junction proteins. Metformin treatment significantly improved glucose tolerance and insulin sensitivity in mice. Moreover, it showed promise in decreasing urinary spot occurrence, reducing urination frequency, alleviating non-voiding contractions, and stabilizing peak urinary pressure. Following metformin therapy, mice displayed restored epithelial fold structure, increased thickness of the muscular layer, substantial decrease in muscle fiber content, notably reduced levels of TXNIP and GSDMD proteins in the metformin-treated group compared to the DBD group, and restored expression of tight junction proteins Zo-1, Claudin-1, and Occludin. Metformin ameliorates urothelial cells damage in DBD mice by inhibiting TXNIP generation and reducing NLRP3 and GSDMD production.

Lactiplantibacillus argentoratensis AGMB00912 alleviates diarrhea and promotes the growth performance of piglets during the weaning transition

BackgroundPreventing post-weaning diarrhea (PWD) in weaned piglets is a crucial challenge in the swine production industry. The stress of weaning, dietary shifts from maternal milk to solid feed, and environmental changes lead to decreased microbial diversity, increased pathogen abundance, and compromised intestinal integrity. We have previously identified Lactiplantibacillus argentoratensis AGMB00912 (LA) in healthy porcine feces, which demonstrated antimicrobial activity against pathogens and enhanced short-chain fatty acid production. This research aimed to evaluate the efficacy of LA strain supplementation as a strategy to inhibit PWD and enhance overall growth performance in weaned piglets.ResultsLA supplementation in weaned piglets significantly increased body weight gain, average daily gain, and average daily feed intake. It also alleviated diarrhea symptoms (diarrhea score and incidence). Notably, LA was found to enrich beneficial microbial populations (Lactobacillus, Anaerobutyricum, Roseburia, Lachnospiraceae, and Blautia) while reducing the abundance of harmful bacteria (Helicobacter and Campylobacter). This not only reduces the direct impact of pathogens but also improves the overall gut microbiota structure, thus enhancing the resilience of weaned piglets. LA treatment also promotes the growth of the small intestinal epithelial structure, strengthens gut barrier integrity, and increases short-chain fatty acid levels in the gut.ConclusionsThe study findings demonstrate the promising potential of LA in preventing PWD. Supplementation with the LA strain offers a promising feed additive for improving intestinal health and growth in piglets during the weaning transition, with the potential to significantly reduce the incidence and severity of PWD.

Drosophila anterior midgut internalization via collective epithelial-mesenchymal transition at the embryo surface and enclosure by surrounding tissues

Internal organ development requires cell internalization, which can occur individually or collectively. The best characterized mode of collective internalization is epithelial invagination. Alternate modes involving collective mesenchymal behaviours at the embryo surface have been documented, but their prevalence is unclear. The Drosophila embryo has been a major model for the study of epithelial invaginations. However, internalization of the Drosophila anterior midgut primordium is incompletely understood. Here, we report that an epithelial-mesenchymal transition (EMT) occurs across the internalizing primordium when it is still at the embryo surface. At the earliest internalization stage, the primordium displays less junctional DE-cadherin than surrounding tissues but still exhibits coordinated epithelial structure as it invaginates with the ventral furrow. This initial invagination is transient, and its loss correlates with the activation of an associated mitotic domain. Activation of a subsequent mitotic domain across the broader primordium results in cell divisions with mixed orientations that deposit some cells within the embryo. However, cell division is non-essential for primordium internalization. Post-mitotically, the surface primordium displays hallmarks of EMT: loss of adherens junctions, loss of epithelial cell polarity, and gain of cell protrusions. Primordium cells extend over each other as they internalize asynchronously as individuals or small groups, and the primordium becomes enclosed by the reorganizations of surrounding epithelial tissues. We propose that collective EMT at the embryo surface promotes anterior midgut internalization through both inwardly-directed divisions and movements of its cells, and that the latter process is facilitated by surrounding tissue remodeling.

Estradiol-mediated enhancement of the human ectocervical epithelial barrier correlates with desmoglein-1 expression in the follicular menstrual phase.

The cervicovaginal epithelial barrier is crucial for defending the female reproductive tract against sexually transmitted infections. Hormones, specifically estradiol and progesterone, along with their respective receptor expressions, play an important role in modulating this barrier. However, the influence of estradiol and progesterone on gene and protein expression in the ectocervical mucosa of naturally cycling women is not well understood. Mucosal and blood samples were collected from Kenyan female sex workers at high risk of sexually transmitted infections. All samples were obtained at two time points, separated by two weeks, aiming for the follicular and luteal phases of the menstrual cycle. Ectocervical tissue biopsies were analyzed by RNA-sequencing and in situ immunofluorescence staining, cervicovaginal lavage samples (CVL) were evaluated using protein profiling, and plasma samples were analyzed for hormone levels. Unsupervised clustering of RNA-sequencing data was performed using Weighted gene co-expression network analysis (WGCNA). In the follicular phase, estradiol levels positively correlated with a gene module representing epithelial structure and function, and negatively correlated with a gene module representing cell cycle regulation. These correlations were confirmed using regression analysis including adjustment for bacterial vaginosis status. Using WGCNA, no gene module correlated with progesterone levels in the follicular phase. In the luteal phase, no gene module correlated with either estradiol or progesterone levels. Protein profiling on CVL revealed that higher levels of estradiol during the follicular phase correlated with increased expression of epithelial barrier integrity markers, including DSG1. This contrasted to the limited correlations of protein expression with estradiol levels in the luteal phase. In situ imaging analysis confirmed that higher estradiol levels during the follicular phase correlated with increased DSG1 expression. We demonstrate that estradiol levels positively correlate with specific markers of ectocervical epithelial structure and function, particularly DSG1, during the follicular phase of the menstrual cycle. Neither progesterone levels during the follicular phase nor estradiol and progesterone levels during the luteal phase correlated with any specific sets of gene markers. These findings align with the expression of estradiol and progesterone receptors in the ectocervical epithelium during these menstrual phases.