RESUMO
BACKGROUND: The osteochondrogenic switch of vascular smooth muscle cells (VSMCs) is a pivotal cellular process in atherosclerotic calcification. However, the exact molecular mechanism of the osteochondrogenic transition of VSMCs remains to be elucidated. Here, we explore the regulatory role of TXNIP (thioredoxin-interacting protein) in the phenotypical transitioning of VSMCs toward osteochondrogenic cells responsible for atherosclerotic calcification. METHODS: The atherosclerotic phenotypes of Txnip-/- mice were analyzed in combination with single-cell RNA-sequencing. The atherosclerotic phenotypes of Tagln-Cre; Txnipflox/flox mice (smooth muscle cell-specific Txnip ablation model), and the mice transplanted with the bone marrow of Txnip-/- mice were analyzed. Public single-cell RNA-sequencing dataset (GSE159677) was reanalyzed to define the gene expression of TXNIP in human calcified atherosclerotic plaques. The effect of TXNIP suppression on the osteochondrogenic phenotypic changes in primary aortic VSMCs was analyzed. RESULTS: Atherosclerotic lesions of Txnip-/- mice presented significantly increased calcification and deposition of collagen content. Subsequent single-cell RNA-sequencing analysis identified the modulated VSMC and osteochondrogenic clusters, which were VSMC-derived populations. The osteochondrogenic cluster was markedly expanded in Txnip-/- mice. The pathway analysis of the VSMC-derived cells revealed enrichment of bone- and cartilage-formation-related pathways and bone morphogenetic protein signaling in Txnip-/- mice. Reanalyzing public single-cell RNA-sequencing dataset revealed that TXNIP was downregulated in the modulated VSMC and osteochondrogenic clusters of human calcified atherosclerotic lesions. Tagln-Cre; Txnipflox/flox mice recapitulated the calcification and collagen-rich atherosclerotic phenotypes of Txnip-/- mice, whereas the hematopoietic deficiency of TXNIP did not affect the lesion phenotype. Suppression of TXNIP in cultured VSMCs accelerates osteodifferentiation and upregulates bone morphogenetic protein signaling. Treatment with the bone morphogenetic protein signaling inhibitor K02288 abrogated the effect of TXNIP suppression on osteodifferentiation. CONCLUSIONS: Our results suggest that TXNIP is a novel regulator of atherosclerotic calcification by suppressing bone morphogenetic protein signaling to inhibit the transition of VSMCs toward an osteochondrogenic phenotype.
Assuntos
Aterosclerose , Calcinose , Placa Aterosclerótica , Calcificação Vascular , Camundongos , Humanos , Animais , Músculo Liso Vascular/metabolismo , Células Cultivadas , Aterosclerose/metabolismo , Placa Aterosclerótica/patologia , Calcinose/metabolismo , Proteínas Morfogenéticas Ósseas/metabolismo , Miócitos de Músculo Liso/metabolismo , RNA/metabolismo , Calcificação Vascular/genética , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Tiorredoxinas/metabolismoRESUMO
Gastric cancer (GC) is histologically classified into intestinal-type gastric cancer (IGC) and diffuse-type gastric cancer (DGC), and the latter is poorly differentiated and highly metastatic. In this study, using quantitative real-time polymerase chain reaction, we described a complete protocol for in vivo CRISPR-Cas9-based knockout screening of essential genes for DGC metastasis. We functionally screened 30 candidate genes using our mouse DGC models lacking Smad4, p53, and E-cadherin. Pooled knockout mouse DGC cells were transplanted into a spleen of syngeneic immunocompetent mice to study clonal advantages in context of a complex process of liver metastasis. Tmsb4x (thymosin beta-4 X-linked), Hmox1, Ifitm3, Ldhb, and Itgb7 were identified as strong candidate genes that promote metastasis. In particular, Tmsb4x enhanced DGC metastasis and stomach organoid-generated tumor growth in in vivo transplantation models. Tmsb4x promoted tumor clonogenicity and anoikis resistance. In situ hybridization analysis showed that Tmsb4x is highly expressed in E-cadherin-negative mouse DGC models compared with mouse IGC and intestinal cancer models. E-cadherin deficiency also increased Tmsb4x expression in stomach organoids via Wnt signaling activation. Collectively, these results demonstrate that Tmsb4x promotes DGC metastasis. In addition, this experimental system will aid in the identification of novel target genes responsible for DGC metastasis.
Assuntos
Biomarcadores Tumorais , Sistemas CRISPR-Cas , Técnicas de Inativação de Genes , Reação em Cadeia da Polimerase em Tempo Real , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologia , Timosina/genética , Animais , Modelos Animais de Doenças , Expressão Gênica , Humanos , Camundongos , Metástase Neoplásica , Transdução de SinaisRESUMO
Canine mammary gland tumors (CMTs) are the most common tumor type in female dogs. This study evaluated the expression pattern and role of thyroglobulin (Tg) in CMT and in human breast cancer (HBC). CMT samples were subjected to fine-needle aspiration, primary cell culture, and histopathology. The expression level of Tg was higher in benign CMT than in malignant CMT (mCMT) primary cells, particularly in the epithelial lineage. Moreover, treatment with Tg enhanced the sensitivity of doxorubicin in mCMT epithelial cells and mitigated proinflammatory response by increasing nuclear factor erythroid 2-related factor 2 (Nrf2). The proximal region of the Tg promoter was hypermethylated in mCMT epithelial cells, silencing Tg expression with concurrent downregulation of Nrf2-mediated antioxidant signaling. An identical pattern of Tg expression was observed in cytological and tissue samples. Tissue microarray analysis showed that Tg was highly expressed in normal and benign tissues when compared with their malignant counterparts, which was diminished along with higher histological grades. The survival rate was significantly higher in HBC patients with high Tg expression than those with low Tg expression. This study also showed that the progression of HBC is accompanied by the reduction of Tg expression along with augmentation of proinflammatory signaling. Our data suggested that Tg could be a negative indicator of malignancy in canine and human breast neoplasia.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/patologia , Doenças do Cão/patologia , Neoplasias Mamárias Animais/patologia , Tireoglobulina/metabolismo , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/mortalidade , Linhagem Celular Tumoral , Doenças do Cão/metabolismo , Cães , Doxorrubicina/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Mamárias Animais/tratamento farmacológico , Neoplasias Mamárias Animais/metabolismo , Metilação , Fator 2 Relacionado a NF-E2/metabolismo , Regiões Promotoras Genéticas , Taxa de Sobrevida , Tireoglobulina/genética , Tireoglobulina/farmacologiaRESUMO
BACKGROUND: Osteopontin (OPN) has been reported to play an important role in intestinal mucosal protection. Although OPN may have positive effects on tight junctions, the exact relationship between OPN and tight junctions has yet to be elucidated. AIMS: To investigate the role of OPN on tight junctions. METHODS: We evaluated clinical signs and histopathology of acute colitis induced by dextran sodium sulfate (DSS) in OPN knockout and wild-type (WT) mice in vivo. Expression levels of occludin and zonula occludens-1 were examined using immunofluorescence. For in vitro analysis, an siRNA-mediated OPN-suppressed Caco-2 monolayer was used. Expression levels and patterns of occludin were analyzed by immunofluorescence, and transepithelial electrical resistance (TER) was measured to evaluate barrier function. Triton X-100 fractionation was used to analyze phosphorylated occludin associated with tight junctional localization. RESULTS: OPN deficiency resulted in an elevated disease activity index, shortened colon length, and aggravated histological signs in mice with DSS-induced acute colitis compared to WT mice. OPN deficiency decreased occludin expression in the colonic mucosa. In Caco-2 monolayers, OPN suppression reduced junctional occludin and redistributed it into the intracellular compartment with decreased TER. Furthermore, western blot for occludin from Triton X-100 insoluble fraction revealed that OPN suppression reduced the phosphorylated form of occludin, which is actually distributed in the tight junction. CONCLUSIONS: Our study showed that OPN is essential for maintaining the tight junction complex by allowing occludin to localize at tight junctions. This could constitute additional evidence that OPN plays a crucial role in intestinal mucosal protection.
Assuntos
Colite/genética , Mucosa Intestinal/patologia , Ocludina/metabolismo , Osteopontina/genética , Junções Íntimas/metabolismo , Animais , Células CACO-2 , Colite/induzido quimicamente , Colite/patologia , Sulfato de Dextrana/toxicidade , Impedância Elétrica , Imunofluorescência , Humanos , Camundongos , Camundongos Knockout , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
Senescence has been demonstrated to either inhibit or promote tumorigenesis. Resolving this paradox requires spatial mapping and functional characterization of senescent cells in the native tumor niche. Here, we identified senescent p16 Ink4a + cancer-associated fibroblasts with a secretory phenotype that promotes fatty acid uptake and utilization by aggressive lung adenocarcinoma driven by Kras and p53 mutations. Furthermore, rewiring of lung cancer metabolism by p16 Ink4a + cancer-associated fibroblasts also altered tumor cell identity to a highly plastic/dedifferentiated state associated with progression in murine and human LUAD. Our ex vivo senolytic screening platform identified XL888, a HSP90 inhibitor, that cleared p16 Ink4a + cancer-associated fibroblasts in vivo. XL888 administration after establishment of advanced lung adenocarcinoma significantly reduced tumor burden concurrent with the loss of plastic tumor cells. Our study identified a druggable component of the tumor stroma that fulfills the metabolic requirement of tumor cells to acquire a more aggressive phenotype.
RESUMO
The accumulation of calcium in atherosclerotic plaques is a prominent feature of advanced atherosclerosis, and it has a strong positive correlation with the total burden of atherosclerosis. Atherosclerotic calcification usually appears first at the necrotic core, indicating that cell death and inflammatory processes are involved in calcification. During atherosclerotic inflammation, various cell types, such as vascular smooth muscle cells, nascent resident pericytes, circulating stem cells, or adventitial cells, have been assumed to differentiate into osteoblastic cells, which lead to vascular calcification. Among these cell types, vascular smooth muscle cells are considered a major contributor to osteochondrogenic cells in the atherosclerotic milieu. In this review, we summarize the molecular mechanisms underlying the osteochondrogenic switch of vascular smooth muscle cells in atherosclerotic plaques.
RESUMO
Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of Johne's disease, a chronic emaciating disease of ruminants that causes enormous economic losses to the bovine industry, globally. However, there are still remaining clues to be solved in the pathogenesis and diagnosis of the disease. Therefore, an in vivo murine experimental model was tried to understand responses in early stage of MAP infection by oral and intraperitoneal (IP) routes. In the MAP infection size, and weight of spleen and liver were increased in the IP group compared with oral groups. Severe histopathological changes were also observed in the spleen and liver of IP infected mice at 12 weeks post-infection (PI). Acid-fast bacterial burden in the organs was closely related to histopathological lesions. In the cytokine production from splenocytes of MAP-infected mice, higher amounts of in TNF-α, IL-10, and IFN-γ were produced at early stage of IP-infected mice while IL-17 production was different at time and infected groups. This phenomenon may indicate the immune shift from Th1 to Th17 through the time course of MAP infection. Systemic and local responses in the MAP-infection were analyzed by using transcriptomic analysis in the spleens and mesenteric lymph nodes (MLN). Based on the analysis of biological processes at 6 weeks PI in spleen and MLN in each infection group, canonical pathways were analyzed with ingenuity pathway analysis in the immune responses and metabolism especially lipid metabolism. Infected host cells with MAP increased in the production of proinflammatory cytokines and reduced the availability of glucose at early stage of infection (p < 0.05). Also, host cells secreted cholesterol through cholesterol efflux to disturb energy source of MAP. These results reveal immunopathological and metabolic responses in the early stage of MAP infection through the development of a murine model.
Assuntos
Doenças dos Bovinos , Mycobacterium avium subsp. paratuberculosis , Paratuberculose , Animais , Bovinos , Camundongos , Modelos Animais de Doenças , Paratuberculose/microbiologia , Citocinas , Colesterol , Doenças dos Bovinos/microbiologiaRESUMO
ST2 functions as a receptor for the cytokine IL-33. It has been implicated in carcinogenesis. In this study, we sought to mechanistically determine how ST2 and IL-33 function to support cancer stem cell (CSC) activity and drive gastric cancer (GC) pathogenesis. ST2+ subpopulation spontaneously arose during gastric tumorigenesis. A thorough evaluation of ST2 and IL-33 expression in gastric tumors revealed that they show an overlapping expression pattern, notably in poor differentiated GC and metastasis foci. Moreover, their expression levels are clinically correlated to cancer progression. Using a genetic model of CSC-driven gastric carcinogenesis, ST2+ subpopulation displays increased tumorigenicity, chemoresistance and metastatic potentials through increased survival fitness endowed by an elevated MAPK-regulated Bcl-xL. The IL-33/ST2 axis enhances the self-renewal and survival of GC stem cells and organoids. Importantly, we observed a synergistic cooperation between IL-33/ST2 and the canonical Wnt pathway in transactivating Wnt-dependent transcription and supporting CSC activity, a partnership that was abrogated by inhibiting Bcl-xL. Concordant with this, ST2+ subpopulation was targeted by MEK1/2 and Bcl-xL-specific inhibitors. These findings establish ST2 as a functional CSC marker that fortifies the Wnt signal while availing a novel therapeutic strategy to suppress GC progression by targeting the IL-33/ST2/Bcl-xL signaling axis.
Assuntos
Neoplasias Gástricas , Via de Sinalização Wnt , Humanos , Neoplasias Gástricas/patologia , Interleucina-33/genética , Interleucina-33/metabolismo , Proteína 1 Semelhante a Receptor de Interleucina-1/genética , Proteína 1 Semelhante a Receptor de Interleucina-1/metabolismo , Carcinogênese/genética , Células-Tronco Neoplásicas/patologia , Linhagem Celular TumoralRESUMO
BACKGROUND: A 10-year-old castrated male Maltese dog was presented with chronic swelling that had been present for at least 5 years in the medial canthus of the right eye (OD). OBJECTIVES: To describe the treatment outcome of dacryops with dacryolithiasis. METHODS: Bilateral patency of the nasolacrimal system was confirmed by flushing of both upper and lower puncta. Ocular ultrasonography revealed a well-defined, oval-shaped, heterogeneous mass with several hyperechoic foci. Dacryocystorhinography revealed no connection between the mass and lacrimal canaliculus. Gentle blunt dissection of the fibrous connective tissue around the cystic mass was performed. The mass was removed, which intraluminally contained multiple calculi. RESULTS: Histopathologically, the cystic structure was lined by simple cuboidal epithelium and surrounded by smooth muscle actin positive myoepithelial cells consistent with dacryops derived from the lacrimal glandular ductal system. In addition, several spherical basophilic minerals were observed in the lumen, which were identified as dacryoliths. CONCLUSION: Surgical removal of this dacryops with dacryolithiasis was curative without recurrence after four months.
Assuntos
Cistos , Doenças do Cão , Doenças do Aparelho Lacrimal , Aparelho Lacrimal , Actinas , Animais , Cistos/cirurgia , Cistos/veterinária , Doenças do Cão/diagnóstico por imagem , Doenças do Cão/cirurgia , Cães , Doenças do Aparelho Lacrimal/cirurgia , Doenças do Aparelho Lacrimal/veterinária , Masculino , MineraisRESUMO
Valvular inflammation triggered by hyperlipidemia has been considered as an important initial process of aortic valve disease; however, cellular and molecular evidence remains unclear. Here, we assess the relationship between plasma lipids and valvular inflammation, and identify association of low-density lipoprotein with increased valvular lipid and macrophage accumulation. Single-cell RNA sequencing analysis reveals the cellular heterogeneity of leukocytes, valvular interstitial cells, and valvular endothelial cells, and their phenotypic changes during hyperlipidemia leading to recruitment of monocyte-derived MHC-IIhi macrophages. Interestingly, we find activated PPARγ pathway in Cd36+ valvular endothelial cells increased in hyperlipidemic mice, and the conservation of PPARγ activation in non-calcified human aortic valves. While the PPARγ inhibition promotes inflammation, PPARγ activation using pioglitazone reduces valvular inflammation in hyperlipidemic mice. These results show that low-density lipoprotein is the main lipoprotein accumulated in the aortic valve during hyperlipidemia, leading to early-stage aortic valve disease, and PPARγ activation protects the aortic valve against inflammation.
Assuntos
Estenose da Valva Aórtica , Calcinose , Hiperlipidemias , Animais , Valva Aórtica/metabolismo , Calcinose/genética , Células Cultivadas , Células Endoteliais/metabolismo , Humanos , Hiperlipidemias/genética , Hiperlipidemias/metabolismo , Imunomodulação , Inflamação/genética , Inflamação/metabolismo , Lipoproteínas LDL/metabolismo , Camundongos , PPAR gama/genética , PPAR gama/metabolismo , Pioglitazona/farmacologia , TranscriptomaRESUMO
Hydrocephalus is one of the most common central nervous system malformations in domestic dogs, yet they are poorly documented and studied in wild carnivoran mammals. A pup of raccoon dog (Nyctereutes procyonoides) was rescued and brought to Wildlife Center. The pup showed generalized ataxia, a domed skull, and an open bregmatic fontanelle. Ultrasound and MRI showed severe enlargement of the lateral ventricle with the loss of septum pellucidum resulting in a single large ventricle and cervical syringohydromyelia. Although treatment was attempted, the animal was euthanized due to poor prognosis. At necropsy, macroscopic findings were identical to the diagnostic imaging, where marked enlargement of the calvarium, and attenuated gyri and sulci were observed. Finally, hydrocephalus was confirmed. Here, we describe a case of hydrocephalus in a raccoon dog (Nyctereutes procyonoides).
Assuntos
Hidrocefalia , Cães Guaxinins , Animais , Animais Selvagens , Hidrocefalia/diagnóstico por imagem , Hidrocefalia/veterináriaRESUMO
BACKGROUND & AIMS: Helicobacter pylori has been reported to modulate local immune responses to colonize persistently in gastric mucosa. Although the induced expression of programmed cell death ligand 1 (PD-L1) has been suggested as an immune modulatory mechanism for persistent infection of H pylori, the main immune cells expressing PD-L1 and their functions in Helicobacter-induced gastritis still remain to be elucidated. METHODS: The blockades of PD-L1 with antibody or PD-L1-deficient bone marrow transplantation were performed in Helicobacter-infected mice. The main immune cells expressing PD-L1 in Helicobacter-infected stomach were determined by flow cytometry and immunofluorescence staining. Helicobacter felis or H pylori-infected dendritic cell (DC)-deficient mouse models including Flt3-/-, Zbtb46-diphtheria toxin receptor, and BDCA2-diphtheria toxin receptor mice were analyzed for pathologic changes and colonization levels. Finally, the location of PD-L1-expressing DCs and the correlation with H pylori infection were analyzed in human gastric tissues using multiplexed immunohistochemistry. RESULTS: Genetic or antibody-mediated blockade of PD-L1 aggravated Helicobacter-induced gastritis with mucosal metaplasia. Gastric classical DCs expressed considerably higher levels of PD-L1 than other immune cells and co-localized with T cells in gastritis lesions from Helicobacter-infected mice and human beings. H felis- or H pylori-infected Flt3-/- or classical DC-depleted mice showed aggravated gastritis with severe T-cell and neutrophil accumulation with low bacterial loads compared with that in control mice. Finally, PD-L1-expressing DCs were co-localized with T cells and showed a positive correlation with H pylori infection in human subjects. CONCLUSIONS: The PD-1/PD-L1 pathway may be responsible for the immune modulatory function of gastric DCs that protects the gastric mucosa from Helicobacter-induced inflammation, but allows persistent Helicobacter colonization.
Assuntos
Antígeno B7-H1/metabolismo , Células Dendríticas/metabolismo , Gastrite/metabolismo , Gastrite/microbiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/fisiologia , Animais , Anticorpos/farmacologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Transplante de Medula Óssea , Antígenos CD11/metabolismo , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/imunologia , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/patologia , Gastrite/patologia , Infecções por Helicobacter/patologia , Helicobacter pylori/efeitos dos fármacos , Inflamação/patologia , Masculino , Metaplasia , Camundongos Endogâmicos C57BL , Linfócitos T/imunologia , Tirosina Quinase 3 Semelhante a fms/deficiência , Tirosina Quinase 3 Semelhante a fms/metabolismoRESUMO
Adipose-derived mesenchymal stromal cells (Ad-MSCs) have excellent potential for skin wound repair. Moreover, platelet-derived growth factor (PDGF) has strong wound healing properties. The purpose of the present study was to compare the healing effects of PDGF-overexpressing canine allogeneic Ad-MSCs (PDGF-MSCs) and their cell sheets (PDGF-CSs) as compared to unexpressed Ad-MSCs (U-MSCs) and their cell sheets (UCSs) in a cutaneous wound healing model induced upon dogs. In in vitro study, the expression of immunomodulatory and growth factors was assessed by qRT-PCR. In in vivo study, cells and sheets were transplanted into a square-shaped full-thickness (1.5×1.5 cm) skin defect model created in 12 dogs. After 5 and 10 days, wounds were harvested and evaluated macroscopically and histopathologically. The qRT-PCR results showed that the PDGF-B gene was significantly upregulated (p<0.05) in PDGF-CS and PDGF-MSCs groups. Upon gross analysis of the wound, all stromal cells and their sheet groups showed accelerated (p<0.05) cutaneous wound healing compared to the negative control groups. As compared to U-MSCs and UCSs, the PDGF-MSCs showed significant epithelization on days 5 and 10 of healing, whereas PDGF-CSs showed improved epithelization only on day 10. In the granulation tissue analysis, PDGF-CSs and UCSs promoted more formation (p<0.05) of upper granulation tissue, collagen, and activated fibroblasts than PDGF-MSCs, and U-MSCs. Especially, the PDGF-CSs presented the highest formation and maturation of granulation tissue among all groups. All considered, PDGF overexpressed stromal cells or cells sheets can improve cutaneous wound healing in a canine model.
Assuntos
Transplante de Células-Tronco Mesenquimais/métodos , Proteínas Proto-Oncogênicas c-sis/biossíntese , Pele , Engenharia Tecidual/métodos , Cicatrização , Animais , Modelos Animais de Doenças , Cães , Lentivirus , Masculino , Células-Tronco Mesenquimais/metabolismo , Pele/lesões , TransfecçãoRESUMO
Here, we report in vivo developmental competent pig embryonic stem cells (ESCs) originating from in vitro-fertilized and parthenogenetic embryos cultured in chemically defined culture media. The pig ESC lines expressed pluripotency markers and were capable of forming teratomas following subcutaneous injection into nude mice. These cell lines would be a useful resource for comparative developmental biology and agricultural biotechnology.
Assuntos
Células-Tronco Embrionárias , Partenogênese , Animais , Diferenciação Celular , Linhagem Celular , Camundongos , Camundongos Nus , SuínosRESUMO
A 3.9-year-old female African pygmy hedgehog (Atelerix albiventris) had a firm, tan-colored mass with an uneven surface arising from the mandibular salivary gland. A histopathologic examination revealed that the mass was composed of neoplastic proliferation of epithelial and spindle cells. The neoplastic spindle cells showed positive for vimentin, smooth muscle actin, calponin and cytokeratin 14 and, negative for cytokeratin 19, suggesting that spindle cells were derived from myoepithelial cells. Based on the histological findings and immunohistochemistry results, the mass was diagnosed as pleomorphic adenoma. Pleomorphic adenoma is the most common benign tumor found in human salivary glands, but it is rare in animals. To the best of our knowledge, this is the first report of pleomorphic adenoma in hedgehogs.
Assuntos
Adenoma Pleomorfo/veterinária , Ouriços , Neoplasias das Glândulas Salivares/veterinária , Adenoma Pleomorfo/patologia , Animais , Feminino , Neoplasias das Glândulas Salivares/patologia , Glândulas Salivares/patologiaRESUMO
Pig embryonic stem cells (pESCs) have been considered an important candidate for preclinical research on human therapies. However, the lack of understanding of pig pluripotent networks has hampered establishment of authentic pESCs. Here, we report that FGF2, ACTVIN, and WNT signaling are essential to sustain pig pluripotency in vitro. Newly derived pESCs were stably maintained over an extended period, and capable of forming teratomas that contained three germ layers. Transcriptome analysis showed that pESCs were developmentally similar to late epiblasts of preimplantation embryos and in terms of biological functions resembled human rather than mouse pluripotent stem cells. However, the pESCs had distinct features such as coexpression of SSEA1 and SSEA4, two active X chromosomes, and a unique transcriptional pattern. Our findings will facilitate both the development of large animal models for human stem cell therapy and the generation of pluripotent stem cells from other domestic animals for agricultural use.
Assuntos
Técnicas de Cultura de Células , Autorrenovação Celular , Meios de Cultura , Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/metabolismo , Animais , Biomarcadores , Blastocisto/citologia , Blastocisto/metabolismo , Diferenciação Celular , Linhagem Celular , Meios de Cultura/química , Citocinas/metabolismo , Citocinas/farmacologia , Feminino , Perfilação da Expressão Gênica , Humanos , Imuno-Histoquímica , Camundongos , SuínosRESUMO
Inflammatory bowel disease (IBD) often accompanies vitamin D deficiency, and vitamin D supplementation ameliorates IBD symptoms in animal models and humans. Because altered vitamin D metabolism has been reported in obesity, we hypothesized that the effects of vitamin D on the development of IBD would be different between obese and control mice. Five-week-old male C57BL/6N mice were divided into 4 groups and fed a diet differing in fat content (10% or 45%, normal diet [ND] or high-fat diet [HFD]) and vitamin D content (1000 or 10â¯000â¯IU/kg of diet, vDC or vDS) for 14â¯weeks. At week 13, colitis was induced by administration of 2% dextran sodium sulfate for 7â¯days. Histology score tended to be lower in the HFD-vDS group than HFD-vDC group, but there was no effect of vitamin D on the ND group. Colonic Cldn1 and Cyp27b1 mRNA levels were higher in the HFD-vDS than HFD-vDC group, but these effects of vitamin D were not observed in the ND group. The serum 25-hydroxy vitamin D levels were negatively correlated with the histology score in the HFD group but not in the ND group. Overall, these results suggest that vitamin D supplementation partially prevents the histological damage of the colon in obese mice but not in control mice. This effect might be mediated by increased colonic Cyp27b1 levels, leading to upregulation of local 1,25-dihydroxy vitamin D production.
Assuntos
Colite/complicações , Colite/tratamento farmacológico , Sulfato de Dextrana/administração & dosagem , Suplementos Nutricionais , Obesidade/complicações , Vitamina D/farmacologia , Animais , Colo/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Vitamina D/administração & dosagem , Vitaminas/administração & dosagem , Vitaminas/farmacologiaRESUMO
A 13-year-old female scimitar-horned oryx (Oryx dammah) died after progressive anorexia, weight loss, and depression. The necropsy showed that the retroperitoneum was compressed by a large white-to-tan uterine mass and on several sections of the mass, the uterine wall was markedly thickened because of ill-defined transmural tumor tissue. Metastatic nodules were detected in the omentum, mesentery, diaphragm, and lung. The genital tract and pulmonary and abdominal nodules exhibited highly pleomorphic sarcoma. The primary and metastatic neoplastic cells showed positive results for vimentin, desmin, and sarcomeric actin, and negative results for smooth muscle actin. Uterine metastatic rhabdomyosarcoma was diagnosed on the basis of the gross, histopathology and immunohistochemistry results.
Assuntos
Rabdomiossarcoma/veterinária , Ruminantes , Neoplasias Uterinas/veterinária , Animais , Feminino , Rabdomiossarcoma/diagnóstico , Rabdomiossarcoma/secundário , Neoplasias Uterinas/diagnósticoRESUMO
Recently, many studies have reported that sensorineural hearing impairment related to neurological disorders may be caused by diabetes mellitus. However, to date, only a small number of studies have investigated the treatment of sensorineural hearing impairment. In the present study, the effects of chlorogenic acid on diabetic auditory pathway impairment were evaluated by neuro-electrical physiological measurements and morphological investigations. We have shown that CA efficiently prevents the progression of auditory pathway dysfunction caused by DM using auditory brainstem responses and auditory middle latency responses in mice. Additionally, using transient-evoked otoacoustic emissions measurement and scanning electron microscope observation of hair cells in DM mice, we found that CA may aid in the recovery from outer hair cell and otic hair cell damage. In conclusion, CA has beneficial effects for the management of diabetic sensorineural auditory dysfunction.
Assuntos
Ácido Clorogênico/farmacologia , Complicações do Diabetes/tratamento farmacológico , Perda Auditiva Neurossensorial/tratamento farmacológico , Hipoglicemiantes/farmacologia , Larva/efeitos dos fármacos , Aloxano , Animais , Apoptose , Vias Auditivas/efeitos dos fármacos , Vias Auditivas/fisiopatologia , Ácido Clorogênico/uso terapêutico , Nervo Coclear/efeitos dos fármacos , Nervo Coclear/fisiopatologia , Complicações do Diabetes/patologia , Complicações do Diabetes/fisiopatologia , Potenciais Evocados Auditivos do Tronco Encefálico/efeitos dos fármacos , Células Ciliadas Auditivas Externas/efeitos dos fármacos , Células Ciliadas Auditivas Externas/patologia , Perda Auditiva Neurossensorial/patologia , Perda Auditiva Neurossensorial/fisiopatologia , Hipoglicemiantes/uso terapêutico , Larva/citologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Peixe-ZebraRESUMO
A six-year-old castrated male Maltese dog presented to a private animal clinic with a mass on the dog's lower lip without any other clinical signs. The mass (3 × 2 × 2 cm) was whitish and grossly well circumscribed, and a histopathological examination revealed that the mass was composed of normal cartilage tissue surrounded by fibrous connective tissues. Based on the gross findings, histopathology and anatomical location of the mass, the first diagnosis of a cartilaginous choristoma in a dog was made.