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1.
Methods Mol Biol ; 2805: 113-124, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39008177

RESUMO

The extracellular matrix (ECM) provides dynamic structural and molecular signals that affect the form and function of developing tissues. In order to parse how the individual features of the ECM impact cell- and tissue-level behavior during development, engineered culture models should reproduce key structural and molecular features of native ECM. Here, we describe a protocol for bioprinting epithelial cell aggregates embedded within a collagen-Matrigel ink in order to study the dynamic interplay between epithelial tissues and aligned networks of type I collagen fibers. Collagen fiber alignment and geometry can be spatially controlled by modulating the printing speed, nozzle geometry, surface chemistry, and degree of molecular crowding in the printing ink. We provide detailed procedures for generating epithelial cell aggregates, microextrusion printing collagen-Matrigel bioinks, culturing the three-dimensional (3D)-printed tissues, and imaging 3D-printed collagen-Matrigel constructs.


Assuntos
Bioimpressão , Colágeno , Células Epiteliais , Matriz Extracelular , Hidrogéis , Impressão Tridimensional , Engenharia Tecidual , Bioimpressão/métodos , Hidrogéis/química , Colágeno/química , Colágeno/metabolismo , Engenharia Tecidual/métodos , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Animais , Morfogênese , Humanos , Proteoglicanas/química , Proteoglicanas/metabolismo , Alicerces Teciduais/química , Laminina/química , Combinação de Medicamentos , Cães , Epitélio/metabolismo , Epitélio/crescimento & desenvolvimento
2.
Cells ; 13(13)2024 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-38994985

RESUMO

The Notch communication pathway, discovered in Drosophila over 100 years ago, regulates a wide range of intra-lineage decisions in metazoans. The division of the Drosophila mechanosensory organ precursor is the archetype of asymmetric cell division in which differential Notch activation takes place at cytokinesis. Here, we review the molecular mechanisms by which epithelial cell polarity, cell cycle and intracellular trafficking participate in controlling the directionality, subcellular localization and temporality of mechanosensitive Notch receptor activation in cytokinesis.


Assuntos
Drosophila melanogaster , Receptores Notch , Animais , Drosophila melanogaster/metabolismo , Receptores Notch/metabolismo , Epitélio/metabolismo , Polaridade Celular , Proteínas de Drosophila/metabolismo , Órgãos dos Sentidos/metabolismo , Órgãos dos Sentidos/citologia , Transdução de Sinais , Células Epiteliais/metabolismo , Células Epiteliais/citologia
3.
Sci Data ; 11(1): 749, 2024 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-38987532

RESUMO

Although early solid diet supplementation is a common practice to improve the growth and development in goat kids, its biological mechanism how solid diet induces rumen microbiota and epithelial development is still unknow. In this study, rumen fermentation parameters, 16S rRNA sequencing for rumen content and epithelial microbiota, transcriptomics and proteomics of epithelium were determined to classify the effects of solid diet supplementation. Here, we classified the changes of goat phenotypes (i.e., growth performance, rumen fermentation and development) and linked them to the changes of rumen microbiota, transcriptome and expressed proteins. The mechanism of solid diet improving rumen development was elucidated preliminarily. Moreover, different roles between the rumen content and epithelial microbiota were identified. Thess datasets expands our understanding of the association between the early diet intervention and rumen development, providing the useful information how nutrient strategy affects rumen function and subsequently improves the host growth. The generated data provides insights in the importance of rumen niche microbiota and microbe-host interactions, which benefits future studies.


Assuntos
Dieta , Cabras , Rúmen , Transcriptoma , Animais , Rúmen/microbiologia , Rúmen/metabolismo , Dieta/veterinária , Ração Animal/análise , Proteômica , Microbioma Gastrointestinal , RNA Ribossômico 16S/genética , Epitélio/metabolismo , Fermentação
4.
Int J Mol Sci ; 25(13)2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-39000205

RESUMO

The ability to precisely treat human disease is facilitated by the sophisticated design of pharmacologic agents. Nanotechnology has emerged as a valuable approach to creating vehicles that can specifically target organ systems, effectively traverse epithelial barriers, and protect agents from premature degradation. In this review, we discuss the molecular basis for epithelial barrier function, focusing on tight junctions, and describe different pathways that drugs can use to cross barrier-forming tissue, including the paracellular route and transcytosis. Unique features of drug delivery applied to different organ systems are addressed: transdermal, ocular, pulmonary, and oral delivery. We also discuss how design elements of different nanoscale systems, such as composition and nanostructured architecture, can be used to specifically enhance transepithelial delivery. The ability to tailor nanoscale drug delivery vehicles to leverage epithelial barrier biology is an emerging theme in the pursuit of facilitating the efficacious delivery of pharmacologic agents.


Assuntos
Sistemas de Liberação de Medicamentos , Nanoestruturas , Humanos , Nanoestruturas/química , Animais , Sistemas de Liberação de Medicamentos/métodos , Junções Íntimas/metabolismo , Transporte Biológico , Epitélio/metabolismo , Células Epiteliais/metabolismo
5.
PLoS One ; 19(6): e0303191, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38924032

RESUMO

BACKGROUND: Gallbladder disease in people is frequently associated with disorders of lipid metabolism and metabolic syndrome. A recently emergent gallbladder disease of dogs, referred to as mucocele formation, is characterized by secretion of abnormal mucus by the gallbladder epithelium and is similarly associated with hyperlipidemia, endocrinopathy, and metabolic dysfunction. The cause of gallbladder mucocele formation in dogs is unknown. METHODS: A prospective case-controlled study was conducted to gain insight into disease pathogenesis by characterization of plasma lipid abnormalities in 18 dogs with gallbladder mucocele formation and 18 age and breed matched control dogs using direct infusion mass spectrometry for complex plasma lipid analysis. This analysis was complemented by histochemical and ultrastructural examination of gallbladder mucosa from dogs with gallbladder mucocele formation and control dogs for evidence of altered lipid homeostasis of the gallbladder epithelium. RESULTS: Gallbladder mucocele formation in dogs carried a unique lipidomic signature of increased lipogenesis impacting 50% of lipid classes, 36% of esterified fatty acid species, and 11% of complex lipid species. Broad enrichment of complex lipids with palmitoleic acid (16:1) and decreased abundance within complex lipids of presumptive omega-3 fatty acids eicosapentaenoic (20:5) and docosahexaenoic (22:6) was significant. Severe lipidosis of gallbladder epithelium pinpoints the gallbladder as involved causally or consequently in abnormal lipid metabolism. CONCLUSION: Our study supports a primary increase in lipogenesis in dogs with mucocele formation and abnormal gallbladder lipid metabolism in disease pathogenesis.


Assuntos
Doenças do Cão , Doenças da Vesícula Biliar , Vesícula Biliar , Lipogênese , Mucocele , Animais , Cães , Mucocele/metabolismo , Mucocele/patologia , Vesícula Biliar/metabolismo , Vesícula Biliar/patologia , Doenças do Cão/metabolismo , Doenças do Cão/patologia , Doenças da Vesícula Biliar/metabolismo , Doenças da Vesícula Biliar/patologia , Doenças da Vesícula Biliar/veterinária , Feminino , Estudos de Casos e Controles , Masculino , Lipidoses/metabolismo , Lipidoses/patologia , Estudos Prospectivos , Epitélio/metabolismo , Epitélio/patologia , Metabolismo dos Lipídeos
6.
Sci Rep ; 14(1): 12744, 2024 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-38830931

RESUMO

Transforming growth factor ß (TGF-ß) is implicated in both mesothelial-to-mesenchymal transition (MMT) and cellular senescence of human peritoneal mesothelial cells (HPMCs). We previously showed that senescent HPMCs could spontaneously acquire some phenotypic features of MMT, which in young HPMCs were induced by TGF-ß. Here, we used electron microscopy, as well as global gene and protein profiling to assess in detail how exposure to TGF-ß impacts on young and senescent HPMCs in vitro. We found that TGF-ß induced structural changes consistent with MMT in young, but not in senescent HPMCs. Of all genes and proteins identified reliably in HPMCs across all treatments and states, 4,656 targets represented overlapping genes and proteins. Following exposure to TGF-ß, 137 proteins and 46 transcripts were significantly changed in young cells, compared to 225 proteins and only 2 transcripts in senescent cells. Identified differences between young and senescent HPMCs were related predominantly to wound healing, integrin-mediated signalling, production of proteases and extracellular matrix components, and cytoskeleton structure. Thus, the response of senescent HPMCs to TGF-ß differs or is less pronounced compared to young cells. As a result, the character and magnitude of the postulated contribution of HPMCs to TGF-ß-induced peritoneal remodelling may change with cell senescence.


Assuntos
Senescência Celular , Células Epiteliais , Peritônio , Fator de Crescimento Transformador beta , Humanos , Senescência Celular/efeitos dos fármacos , Fator de Crescimento Transformador beta/metabolismo , Fator de Crescimento Transformador beta/farmacologia , Células Epiteliais/metabolismo , Células Epiteliais/efeitos dos fármacos , Peritônio/citologia , Peritônio/metabolismo , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Células Cultivadas , Epitélio/metabolismo , Epitélio/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Perfilação da Expressão Gênica
7.
Cell ; 187(12): 2898-2900, 2024 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-38848672

RESUMO

Epithelial folding is a fundamental biological process that requires epithelial interactions with the underlying mesenchyme. In this issue of Cell, Huycke et al. investigate intestinal villus formation. They discover that water-droplet-like behavior of mesenchymal cells drives their coalescence into uniformly patterned aggregates, which generate forces on the epithelium to initiate folding.


Assuntos
Epitélio , Mesoderma , Animais , Humanos , Células Epiteliais/metabolismo , Células Epiteliais/citologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/citologia , Mesoderma/metabolismo , Mesoderma/citologia , Epitélio/metabolismo
8.
Sci Robot ; 9(91): eadl2007, 2024 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-38924422

RESUMO

Cytokines have been identified as key contributors to the development of inflammatory bowel disease (IBD), yet conventional treatments often prove inadequate and carry substantial side effects. Here, we present an innovative biohybrid robotic system, termed "algae-MΦNP-robot," for addressing IBD by actively neutralizing colonic cytokine levels. Our approach combines moving green microalgae with macrophage membrane-coated nanoparticles (MΦNPs) to efficiently capture proinflammatory cytokines "on the fly." The dynamic algae-MΦNP-robots outperformed static counterparts by enhancing cytokine removal through continuous movement, better distribution, and extended retention in the colon. This system is encapsulated in an oral capsule, which shields it from gastric acidity and ensures functionality upon reaching the targeted disease site. The resulting algae-MΦNP-robot capsule effectively regulated cytokine levels, facilitating the healing of damaged epithelial barriers. It showed markedly improved prevention and treatment efficacy in a mouse model of IBD and demonstrated an excellent biosafety profile. Overall, our biohybrid algae-MΦNP-robot system offers a promising and efficient solution for IBD, addressing cytokine-related inflammation effectively.


Assuntos
Colo , Citocinas , Doenças Inflamatórias Intestinais , Nanopartículas , Robótica , Animais , Citocinas/metabolismo , Doenças Inflamatórias Intestinais/metabolismo , Robótica/instrumentação , Camundongos , Humanos , Macrófagos/metabolismo , Mucosa Intestinal/metabolismo , Modelos Animais de Doenças , Camundongos Endogâmicos C57BL , Masculino , Desenho de Equipamento , Epitélio
9.
J Nanobiotechnology ; 22(1): 331, 2024 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-38867284

RESUMO

BACKGROUND: In the context of increasing exposure to silica nanoparticles (SiNPs) and ensuing respiratory health risks, emerging evidence has suggested that SiNPs can cause a series of pathological lung injuries, including fibrotic lesions. However, the underlying mediators in the lung fibrogenesis caused by SiNPs have not yet been elucidated. RESULTS: The in vivo investigation verified that long-term inhalation exposure to SiNPs induced fibroblast activation and collagen deposition in the rat lungs. In vitro, the uptake of exosomes derived from SiNPs-stimulated lung epithelial cells (BEAS-2B) by fibroblasts (MRC-5) enhanced its proliferation, adhesion, and activation. In particular, the mechanistic investigation revealed SiNPs stimulated an increase of epithelium-secreted exosomal miR-494-3p and thereby disrupted the TGF-ß/BMPR2/Smad pathway in fibroblasts via targeting bone morphogenetic protein receptor 2 (BMPR2), ultimately resulting in fibroblast activation and collagen deposition. Conversely, the inhibitor of exosomes, GW4869, can abolish the induction of upregulated miR-494-3p and fibroblast activation in MRC-5 cells by the SiNPs-treated supernatants of BEAS-2B. Besides, inhibiting miR-494-3p or overexpression of BMPR2 could ameliorate fibroblast activation by interfering with the TGF-ß/BMPR2/Smad pathway. CONCLUSIONS: Our data suggested pulmonary epithelium-derived exosomes serve an essential role in fibroblast activation and collagen deposition in the lungs upon SiNPs stimuli, in particular, attributing to exosomal miR-494-3p targeting BMPR2 to modulate TGF-ß/BMPR2/Smad pathway. Hence, strategies targeting exosomes could be a new avenue in developing therapeutics against lung injury elicited by SiNPs.


Assuntos
Colágeno , Epigênese Genética , Exossomos , Fibroblastos , Pulmão , MicroRNAs , Nanopartículas , Transdução de Sinais , Dióxido de Silício , Fator de Crescimento Transformador beta , Exossomos/metabolismo , Animais , Fibroblastos/metabolismo , Fibroblastos/efeitos dos fármacos , Dióxido de Silício/química , Transdução de Sinais/efeitos dos fármacos , Ratos , Pulmão/metabolismo , Pulmão/patologia , Colágeno/metabolismo , Humanos , Nanopartículas/química , MicroRNAs/metabolismo , MicroRNAs/genética , Linhagem Celular , Fator de Crescimento Transformador beta/metabolismo , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/induzido quimicamente , Masculino , Células Epiteliais/metabolismo , Células Epiteliais/efeitos dos fármacos , Ratos Sprague-Dawley , Epitélio/metabolismo , Epitélio/efeitos dos fármacos
10.
J Hazard Mater ; 475: 134900, 2024 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-38878440

RESUMO

The expected increments in the production/use of bioplastics, as an alternative to petroleum-based plastics, require a deep understanding of their potential environmental and health hazards, mainly as nanoplastics (NPLs). Since one important exposure route to NPLs is through inhalation, this study aims to determine the fate and effects of true-to-life polylactic acid nanoplastics (PLA-NPLs), using the in vitro Calu-3 model of bronchial epithelium, under air-liquid interphase exposure conditions. To determine the harmful effects of PLA-NPLs in a more realistic scenario, both acute (24 h) and long-term (1 and 2 weeks) exposures were used. Flow cytometry results indicated that PLA-NPLs internalized easily in the barrier (∼10 % at 24 h and ∼40 % after 2 weeks), which affected the expression of tight-junctions formation (∼50 % less vs control) and the mucus secretion (∼50 % more vs control), both measured by immunostaining. Interestingly, significant genotoxic effects (DNA breaks) were detected by using the comet assay, with long-term effects being more marked than acute ones (7.01 vs 4.54 % of DNA damage). When an array of cellular proteins including cytokines, chemokines, and growth factors were used, a significant over-expression was mainly found in long-term exposures (∼20 proteins vs 5 proteins after acute exposure). Overall, these results described the potential hazards posed by PLA-NPLs, under relevant long-term exposure scenarios, highlighting the advantages of the model used to study bronchial epithelium tissue damage, and signaling endpoints related to inflammation.


Assuntos
Poliésteres , Poliésteres/toxicidade , Poliésteres/química , Humanos , Linhagem Celular , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Citocinas/metabolismo , Microplásticos/toxicidade , Dano ao DNA/efeitos dos fármacos , Nanopartículas/toxicidade , Nanopartículas/química , Epitélio/efeitos dos fármacos , Mucosa Respiratória/efeitos dos fármacos , Mucosa Respiratória/metabolismo , Células Epiteliais/efeitos dos fármacos , Junções Íntimas/efeitos dos fármacos
11.
Cell Rep ; 43(6): 114271, 2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38823013

RESUMO

The epithelial adaptations to mechanical stress are facilitated by molecular and tissue-scale changes that include the strengthening of junctions, cytoskeletal reorganization, and cell-proliferation-mediated changes in tissue rheology. However, the role of cell size in controlling these properties remains underexplored. Our experiments in the zebrafish embryonic epidermis, guided by theoretical estimations, reveal a link between epithelial mechanics and cell size, demonstrating that an increase in cell size compromises the tissue fracture strength and compliance. We show that an increase in E-cadherin levels in the proliferation-deficient epidermis restores epidermal compliance but not the fracture strength, which is largely regulated by Ezrin-an apical membrane-cytoskeleton crosslinker. We show that Ezrin fortifies the epithelium in a cell-size-dependent manner by countering non-muscle myosin-II-mediated contractility. This work uncovers the importance of cell size maintenance in regulating the mechanical properties of the epithelium and fostering protection against future mechanical stresses.


Assuntos
Tamanho Celular , Proteínas do Citoesqueleto , Miosina Tipo II , Peixe-Zebra , Animais , Peixe-Zebra/metabolismo , Proteínas do Citoesqueleto/metabolismo , Proteínas do Citoesqueleto/genética , Miosina Tipo II/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genética , Estresse Mecânico , Células Epiteliais/metabolismo , Caderinas/metabolismo , Epiderme/metabolismo , Epitélio/metabolismo , Proliferação de Células
12.
J Anim Sci ; 1022024 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-38902909

RESUMO

The aim of this study was to investigate the effects of ensiled agricultural byproducts from Qinghai-Tibet plateau on growth performance, rumen microbiota, ruminal epithelium morphology, and nutrient transport-related gene expression in Tibetan sheep. Fourteen male Tibetan sheep were randomly assigned to one of two diets: an untreated diet (without silage inoculum, CON, n = 7) or an ensiled diet (with silage inoculum, ESD, n = 7). The total experimental period lasted for 84 d, including early 14 d as adaption period and remaining 70 d for data collection. The ESD increased average daily gain (P = 0.046), dry matter intake (P < 0.001), ammonia nitrogen (P = 0.045), microbial crude protein (P = 0.034), and total volatile fatty acids concentration (P < 0.001), and decreased ruminal pH value (P = 0.014). The proportion of propionate (P = 0.006) and the copy numbers of bacteria (P = 0.01) and protozoa (P = 0.002) were higher, while the proportion of acetate (P = 0.028) was lower in the sheep fed ESD compared to CON. Pyrosequencing of the 16S ribosomal RNA gene revealed that ESD increased the relative abundance of Firmicutes, Ruminococcus, Lachnospiraceae_AC2044_group, Lachnospiraceae_XPB1014_group, and Christensenellaceae_R-7_group in the rumen (P < 0.05), while decreased the relative abundance of Bacteroidota, Prevotellaceae_UCG-003, and Veillonellaceae_UCG-001 (P < 0.05). Analyses with PICRUSt2 and STAMP indicated that the propionate metabolism pathway was enriched in the sheep fed ESD (P = 0.026). The ESD increased the rumen papillae height (P = 0.012), density (P = 0.036), and surface area (P = 0.001), and improved the thickness of the total epithelia (P = 0.018), stratum corneum (P = 0.040), stratum granulosum (P = 0.042), and stratum spinosum and basale (P = 0.004). The relative mRNA expression of cyclin-dependent Kinase 2, CyclinA2, CyclinD2, zonula occludens-1, Occludin, monocarboxylate transporter isoform 1 (MCT1), MCT4, sodium/potassium pump, and sodium/hydrogen antiporter 3 were higher in the rumen epithelial of sheep fed ESD than CON (P < 0.05). Conversely, the relative mRNA expressions of Caspase 3 and B-cell lymphoma-2 were lower in the sheep fed ESD than CON (P < 0.05). In conclusion, compared with an untreated diet, feeding an ensiled diet altered the rumen microbial community, enhanced nutrient transport through rumen epithelium, and improved the growth performance of Tibetan sheep.


Tibetan sheep on the Qinghai-Tibet Plateau experience significant nutrient stress while a substantial amount of agricultural byproducts in the region go discarded and wasted. In this study, agricultural byproducts were ensiled and fed to the Tibetan sheep to investigate their effects on growth performance, rumen microorganisms, and nutrient transport through rumen epithelial tissues. Fourteen male Tibetan sheep were randomly assigned to one of two diets: untreated diet (without silage inoculum, CON, n = 7) or ensiled diet (with silage inoculum, ESD, n = 7). After 70 d of feeding, the ESD-fed sheep had a higher body weight than CON. The ensiled diet changed the rumen microbial community and increased the relative abundance of cellulolytic bacteria in the rumen. In addition, the ensiled diet also promoted the development of rumen epithelia and improved the relative expression of gene related to nutrient transport. Overall, the ensiled diet optimized the use of agricultural byproducts and significantly contributed to the production of Tibetan sheep.


Assuntos
Ração Animal , Dieta , Rúmen , Silagem , Animais , Rúmen/microbiologia , Ovinos/fisiologia , Ovinos/crescimento & desenvolvimento , Masculino , Dieta/veterinária , Ração Animal/análise , Silagem/análise , Tibet , Microbioma Gastrointestinal/efeitos dos fármacos , Epitélio , Fenômenos Fisiológicos da Nutrição Animal , Distribuição Aleatória , Bactérias/classificação
13.
Int J Mol Sci ; 25(11)2024 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-38891804

RESUMO

The periodontium comprising periodontal ligament (PDL), gingiva, and epithelium play crucial roles in maintaining tooth integrity and function. Understanding tissue cellular composition and gene expression is crucial for illuminating periodontal pathophysiology. This study aimed to identify tissue-specific markers via scRNA-Seq. Primary human PDL, gingiva, and epithelium tissues (n = 7) were subjected to cell hashing and sorting. scRNA-Seq library preparation using 10× Genomics protocol and Illumina sequencing was conducted. The analysis was performed using Cellranger (v3.1.0), with downstream analysis via R packages Seurat (v5.0.1) and SCORPIUS (v1.0.9). Investigations identified eight distinct cellular clusters, revealing the ubiquitous presence of epithelial and gingival cells. PDL cells evolved in two clusters with numerical superiority. The other clusters showed varied predominance regarding gingival and epithelial cells or an equitable distribution of both. The cluster harboring most cells mainly consisted of PDL cells and was present in all donors. Some of the other clusters were also tissue-inherent, while the presence of others was environmentally influenced, revealing variability across donors. Two clusters exhibited genetic profiles associated with tissue development and cellular integrity, respectively, while all other clusters were distinguished by genes characteristic of immune responses. Developmental trajectory analysis uncovered that PDL cells may develop after epithelial and gingival cells, suggesting the inherent PDL cell-dominated cluster as a final developmental stage. This single-cell RNA sequencing study delineates the hierarchical organization of periodontal tissue development, identifies tissue-specific markers, and reveals the influence of environmental factors on cellular composition, advancing our understanding of periodontal biology and offering potential insights for therapeutic interventions.


Assuntos
Gengiva , Ligamento Periodontal , Análise de Célula Única , Transcriptoma , Humanos , Ligamento Periodontal/metabolismo , Ligamento Periodontal/citologia , Gengiva/metabolismo , Análise de Célula Única/métodos , Análise de Sequência de RNA/métodos , Perfilação da Expressão Gênica , Epitélio/metabolismo , Células Epiteliais/metabolismo , Feminino , Masculino
14.
PLoS Genet ; 20(6): e1011326, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38857279

RESUMO

The development of ectodermal organs begins with the formation of a stratified epithelial placode that progressively invaginates into the underlying mesenchyme as the organ takes its shape. Signaling by secreted molecules is critical for epithelial morphogenesis, but how that information leads to cell rearrangement and tissue shape changes remains an open question. Using the mouse dentition as a model, we first establish that non-muscle myosin II is essential for dental epithelial invagination and show that it functions by promoting cell-cell adhesion and persistent convergent cell movements in the suprabasal layer. Shh signaling controls these processes by inducing myosin II activation via AKT. Pharmacological induction of AKT and myosin II can also rescue defects caused by the inhibition of Shh. Together, our results support a model in which the Shh signal is transmitted through myosin II to power effective cellular rearrangement for proper dental epithelial invagination.


Assuntos
Adesão Celular , Movimento Celular , Proteínas Hedgehog , Miosina Tipo II , Transdução de Sinais , Animais , Camundongos , Proteínas Hedgehog/metabolismo , Proteínas Hedgehog/genética , Adesão Celular/genética , Miosina Tipo II/metabolismo , Miosina Tipo II/genética , Movimento Celular/genética , Epitélio/metabolismo , Morfogênese/genética , Dente/metabolismo , Dente/crescimento & desenvolvimento , Células Epiteliais/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Regulação da Expressão Gênica no Desenvolvimento
15.
Ultrasound Med Biol ; 50(8): 1247-1254, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38834492

RESUMO

OBJECTIVE: Needle biopsy is a common technique used to obtain cell and tissue samples for diagnostics. Currently, two biopsy methods are widely used: (i) fine-needle aspiration biopsy (FNAB) and (ii) core needle biopsy (CNB). However, these methods have limitations. Recently, we developed ultrasound-enhanced fine-needle aspiration biopsy (USeFNAB), which employs a needle that flexurally oscillates at an ultrasonic frequency of ∼32 kHz. The needle motion contributes to increased tissue collection while preserving cells and tissue constructs for pathological assessment. Previously, USeFNAB has been investigated only in ex vivo animal tissue. The present study was aimed at determining the feasibility of using USeFNAB in human epithelial and lymphoid tissue. METHODS: Needle biopsy samples were acquired using FNAB, CNB and USeFNAB on ex vivo human tonsils (N = 10). The tissue yield and quality were quantified by weight measurement and blinded pathologists' assessments. The biopsy methods were then compared. RESULTS: The results revealed sample mass increases of, on average, 2.3- and 5.4-fold with USeFNAB compared with the state-of-the-art FNAB and CNB, respectively. The quality of tissue fragments collected by USeFNAB was equivalent to that collected by the state-of-the-art methods in terms of morphology and immunohistochemical stainings made from cell blocks as judged by pathologists. CONCLUSION: Our study indicates that USeFNAB is a promising method that could improve tissue yield to ensure sufficient material for ancillary histochemical and molecular studies for diagnostic pathology, thereby potentially increasing diagnostic accuracy.


Assuntos
Tecido Linfoide , Tonsila Palatina , Humanos , Tonsila Palatina/patologia , Tonsila Palatina/diagnóstico por imagem , Tecido Linfoide/patologia , Tecido Linfoide/diagnóstico por imagem , Biópsia por Agulha Fina/métodos , Estudos de Viabilidade , Ultrassonografia de Intervenção/métodos , Biópsia Guiada por Imagem/métodos , Epitélio/patologia
16.
Pathol Res Pract ; 258: 155320, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38728794

RESUMO

The objective of this study to examine the effects of curcumin and gallic acid use against oxidative stress damage in the autologous intraperitoneal ovarian transplantation model created in rats on ovarian follicle reserve, ovarian surface epithelium, and oxidant-antioxidant systems. 42 adult female Sprague Dawley rats (n=7) were allocated into 6 groups. Group 1 served as the control. In Group 2, rats underwent ovarian transplantation (TR) to their peritoneal walls. Group 3 received corn oil (CO) (0.5 ml/day) one day before and 14 days after transplantation. Group 4 was administered curcumin (CUR) (100 mg/kg/day), Group 5 received gallic acid (GA) (20 mg/kg/day), and Group 6 was treated with a combination of curcumin and gallic acid via oral gavage after transplantation. Rats were sacrificed on the 14th postoperative day, and blood along with ovaries were collected for analysis. The removed ovaries were analyzed at light microscopic, fluorescence microscopic, and biochemical levels. In Group 2 and Group 3, while serum and tissue Total Oxidant Levels (TOS) and Oxidative Stress Index (OSI) increased, serum Total Antioxidant Levels (TAS) decreased statistically significantly (p˂0.05) compared to the other groups (Groups 1, 4, 5, and 6). The ovarian follicle reserve was preserved and the changes in the ovarian surface epithelium and histopathological findings were reduced in the antioxidant-treated groups (Groups 4, 5, and 6). In addition, immunofluorescence examination revealed that the expression of Cytochrome C and Caspase 3 was stronger and Ki-67 was weaker in Groups 2 and 3, in comparison to the groups that were given antioxidants. It can be said that curcumin and gallic acid have a histological and biochemical protective effect against ischemia-reperfusion injury due to ovarian transplantation, and this effect is stronger when these two antioxidants are applied together compared to individual use.


Assuntos
Antioxidantes , Curcumina , Ácido Gálico , Folículo Ovariano , Reserva Ovariana , Ovário , Estresse Oxidativo , Ratos Sprague-Dawley , Animais , Feminino , Ácido Gálico/farmacologia , Curcumina/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Ovário/efeitos dos fármacos , Ovário/patologia , Ovário/metabolismo , Ratos , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/metabolismo , Folículo Ovariano/patologia , Reserva Ovariana/efeitos dos fármacos , Antioxidantes/farmacologia , Epitélio/efeitos dos fármacos , Epitélio/patologia , Epitélio/metabolismo , Transplante Autólogo , Sinergismo Farmacológico
17.
J Med Invest ; 71(1.2): 29-39, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38735722

RESUMO

The establishment of an adaptive immune system is critical for protecting our bodies from neoplastic cancers and invading pathogens such as viruses and bacteria. As a primary lymphoid organ, the thymus generates lymphoid T cells that play a major role in the adaptive immune system. T cell generation in the thymus is controlled by interactions between thymocytes and other thymic cells, primarily thymic epithelial cells. Thus, the normal development and function of thymic epithelial cells are important for the generation of immunocompetent and self-tolerant T cells. On the other hand, the degeneration of the thymic epithelium due to thymic aging causes thymic involution, which is associated with the decline of adaptive immune function. Herein we summarize basic and current knowledge of the development and function of thymic epithelial cells and the mechanism of thymic involution. J. Med. Invest. 71 : 29-39, February, 2024.


Assuntos
Envelhecimento , Timo , Timo/imunologia , Timo/crescimento & desenvolvimento , Humanos , Envelhecimento/fisiologia , Envelhecimento/imunologia , Animais , Células Epiteliais/fisiologia , Epitélio/imunologia , Linfócitos T/imunologia
19.
Nat Commun ; 15(1): 3756, 2024 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-38704381

RESUMO

The human pathogen Neisseria gonorrhoeae ascends into the upper female reproductive tract to cause damaging inflammation within the Fallopian tubes and pelvic inflammatory disease (PID), increasing the risk of infertility and ectopic pregnancy. The loss of ciliated cells from the epithelium is thought to be both a consequence of inflammation and a cause of adverse sequelae. However, the links between infection, inflammation, and ciliated cell extrusion remain unresolved. With the use of ex vivo cultures of human Fallopian tube paired with RNA sequencing we defined the tissue response to gonococcal challenge, identifying cytokine, chemokine, cell adhesion, and apoptosis related transcripts not previously recognized as potentiators of gonococcal PID. Unexpectedly, IL-17C was one of the most highly induced genes. Yet, this cytokine has no previous association with gonococcal infection nor pelvic inflammatory disease and thus it was selected for further characterization. We show that human Fallopian tubes express the IL-17C receptor on the epithelial surface and that treatment with purified IL-17C induces pro-inflammatory cytokine secretion in addition to sloughing of the epithelium and generalized tissue damage. These results demonstrate a previously unrecognized but critical role of IL-17C in the damaging inflammation induced by gonococci in a human explant model of PID.


Assuntos
Tubas Uterinas , Gonorreia , Inflamação , Interleucina-17 , Neisseria gonorrhoeae , Adulto , Feminino , Humanos , Citocinas/metabolismo , Epitélio/patologia , Epitélio/microbiologia , Tubas Uterinas/microbiologia , Tubas Uterinas/patologia , Tubas Uterinas/imunologia , Gonorreia/imunologia , Gonorreia/microbiologia , Gonorreia/patologia , Inflamação/patologia , Inflamação/microbiologia , Interleucina-17/metabolismo , Neisseria gonorrhoeae/imunologia , Neisseria gonorrhoeae/patogenicidade , Doença Inflamatória Pélvica/microbiologia , Doença Inflamatória Pélvica/patologia , Doença Inflamatória Pélvica/imunologia , Receptores de Interleucina-17/metabolismo , Receptores de Interleucina-17/genética
20.
Life Sci ; 348: 122695, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38710285

RESUMO

AIMS: To evaluate the basal release of 6-nitrodopamine (6-ND) from human isolated seminal vesicles (HISV) and to characterize its action and origin. MAIN METHODS: Left HISV obtained from patients undergoing prostatectomy surgery was suspended in a 3-mL organ bath containing warmed (37 °C) and gassed (95%O2:5%CO2) Krebs-Henseleit's solution (KHS) with ascorbic acid. An aliquot of 2 mL of the supernatant was used to quantify catecholamines by LC-MS/MS. For functional studies, concentration-responses curves to catecholamines were obtained, and pEC50 and Emax values were calculated. Detection of tyrosine hydroxylase and S100 protein were also carried out by both immunohistochemistry and fluorescence in-situ hybridization assays (FISH). KEY FINDINGS: Basal release of 6-ND was higher than the other catecholamines (14.76 ± 14.54, 4.99 ± 6.92, 3.72 ± 4.35 and 5.13 ± 5.76 nM for 6-ND, noradrenaline, adrenaline, and dopamine, respectively). In contrast to the other catecholamines, the basal release of 6-ND was not affected by the sodium current (Nav) channel inhibitor tetrodotoxin (1 µM; 10.4 ± 8.9 and 10.4 ± 7.9 nM, before and after tetrodotoxin, respectively). All the catecholamines produced concentration-dependent HISV contractions (pEC50 4.1 ± 0.2, 4.9 ± 0.3, 5.0 ± 0.3, and 3.9 ± 0.8 for 6-ND, noradrenaline, adrenaline, and dopamine, respectively), but 6-ND was 10-times less potent than noradrenaline and adrenaline. However, preincubation with very low concentration of 6-ND (10-8 M, 30 min) produced significant leftward shifts of the concentration-response curves to noradrenaline. Immunohistochemical and FISH assays identified tyrosine hydroxylase in tissue epithelium of HISV strips. SIGNIFICANCE: Epithelium-derived 6-ND is the major catecholamine released from human isolated seminal vesicles and that modulates smooth muscle contractility by potentiating noradrenaline-induced contractions.


Assuntos
Dopamina , Norepinefrina , Glândulas Seminais , Humanos , Masculino , Norepinefrina/farmacologia , Norepinefrina/metabolismo , Glândulas Seminais/efeitos dos fármacos , Glândulas Seminais/metabolismo , Dopamina/metabolismo , Dopamina/farmacologia , Pessoa de Meia-Idade , Epitélio/metabolismo , Epitélio/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Idoso , Catecolaminas/metabolismo
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