RESUMO
Inflammation is involved in the induction of chronic inflammatory and neuropathic pain. Moreover, the ketogenic diet, a high-fat, low-carbohydrate, and adequate protein diet, has an anti-inflammatory effect. Thus, we hypothesized that a ketogenic diet has a therapeutic effect on both types of chronic pain. In the present study, we investigated the effect of a ketogenic diet on mechanical allodynia, a chronic pain symptom, in formalin-induced chronic inflammatory pain and nerve injury-induced neuropathic pain models using adult male mice. Formalin injection into the hind paw induced mechanical allodynia in both the injected and intact hind paws, and the ketogenic diet alleviated mechanical allodynia in both hind paws. In addition, the ketogenic diet prevented formalin-induced edema. Furthermore, the diet alleviated mechanical allodynia induced by peripheral nerve injury. Thus, these findings indicate that a ketogenic diet has a therapeutic effect on chronic pain induced by inflammation and nerve injury.
Assuntos
Dor Crônica , Dieta Cetogênica , Neuralgia , Camundongos , Masculino , Animais , Hiperalgesia/tratamento farmacológico , Inflamação/metabolismo , Formaldeído/efeitos adversos , Modelos Animais de DoençasRESUMO
DNA methylation of cytosine bases is a major epigenetic modification that regulates gene expression and vertebrate development. The ten-eleven translocation (TET) enzymes oxidize 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), and active DNA demethylation influences gene expression specific to each developmental stage, although recent reports have shown that TET also has a non-catalytic function. In fetal mice, the epithelium in the submandibular gland (SMG) buds as a derivative of the oral cavity at embryonic day 11 (E11) and, by E15, it begins to differentiate into the salivary epithelium, which expresses water-channel aquaporin 5 (AQP5). The functional differentiation of the SMG epithelium can be regulated epigenetically, but how TET enzymes contribute is largely unknown. Here, we used several techniques, including hydroxymethylated DNA immunoprecipitation qPCR and histological analysis, to examine the changes in 5hmC levels and AQP5 and TET expression during SMG development. We found that 5hmC levels and AQP5 expression increased in the E15 SMG epithelium, while TET2 expression in the terminal buds decreased at E15. In agreement with the in vivo observations, Tet2 inhibition ex vivo led to the upregulation of AQP5 expression in terminal buds of the SMG epithelium. These results suggest that the downregulation of TET2 expression at E15 is a critical epigenetic event that establishes the epithelial fate for functional SMGs during development.
Assuntos
Dioxigenases , 5-Metilcitosina/metabolismo , Animais , Metilação de DNA , Proteínas de Ligação a DNA/genética , Dioxigenases/genética , Dioxigenases/metabolismo , Regulação para Baixo/genética , Camundongos , Organogênese/genética , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismoRESUMO
We analyzed the clinical features of 157 patients admitted to 11 institutions in the Kagawa Prefecture for volvulus of the sigmoid colon. The following were the background information of the patients:median age, 79.0 years;male-to-female ratio, 102:55;median body mass index, 20.0kg/m2;and the proportion of patients with performance status ≥3, 43.9%. Abdominal bloating and pain were the chief complaints. During hospitalization, endoscopy and endoscopic detorsion were performed 157 and 100 times, respectively. An accidental complication was observed in 3 cases, all of which were intestinal perforations. Surgery, which was indicated for ischemia, was performed in 62 of the 157 cases. Endoscopy is useful in the diagnosis of ischemia, which can be treated following an early diagnosis. Of the 157 patients, 19 died, whereas the rest were discharged. The risk factors for death were age ≥80 years and creatinine kinase level ≥200IU/L.
Assuntos
Volvo Intestinal , Doenças do Colo Sigmoide , Idoso , Idoso de 80 Anos ou mais , Colo Sigmoide , Endoscopia , Feminino , Hospitais , Humanos , Volvo Intestinal/diagnóstico por imagem , Volvo Intestinal/epidemiologia , Volvo Intestinal/cirurgia , Masculino , Estudos Retrospectivos , Doenças do Colo Sigmoide/epidemiologia , Doenças do Colo Sigmoide/cirurgiaRESUMO
Efficient, earth-abundant, and acid-stable catalysts for the oxygen evolution reaction (OER) are missing pieces for the production of hydrogen via water electrolysis. Here, we report how the limitations on the stability of 3d-metal materials can be overcome by the spectroscopic identification of stable potential windows in which the OER can be catalyzed efficiently while simultaneously suppressing deactivation pathways. We demonstrate the benefits of this approach using gamma manganese oxide (γ-MnO2 ), which shows no signs of deactivation even after 8000â h of electrolysis at a pH of 2. This stability is vastly superior to existing acid-stable 3d-metal OER catalysts, but cannot be realized if there is a deviation as small as 50-mV from the stable potential window. A stable voltage efficiency of over 70 % in a polymer-electrolyte membrane (PEM) electrolyzer further verifies the availability of this approach and showcases how materials previously perceived to be unstable may have potential application for water electrolysis in an acidic environment.
RESUMO
The hedgehog family includes Sonic hedgehog (Shh), Desert hedgehog, and Indian hedgehog, which are well known as a morphogens that play many important roles during development of numerous organs such as the tongue, pancreas, kidney, cartilage, teeth and salivary glands (SMG). In Shh null mice, abnormal development of the salivary gland is seen after embryonic day 14 (E14). Shh also induced lobule formation and lumen formation in acini-like structures in cultured E14 SMG. In this study, we investigated the relationship between Shh and epidermal growth factor (EGF)/ErbB signaling in developing fetal mouse SMG. Administration of Shh to cultured E13 SMG stimulated branching morphogenesis (BrM) and induced synthesis of mRNAs for EGF ligands and receptors of the ErbB family. Shh also stimulated activation of ErbB signaling system such as ERK1/2. AG1478, a specific inhibitor of ErbB receptors, completely suppressed BrM and activation of EGF/ErbB/ERK1/2 cascade in E13 SMGs cultured with Shh. The expressions of mRNA for Egf in mesenchyme and mRNA for Erbb1, Erbb2 and Erbb3 in epithelium of E13 SMG were specifically induced by administration of Shh. These results show that Shh stimulates BrM of fetal mouse SMG, at least in part, through activation of the EGF/ErbB/ERK1/2 signaling system.
Assuntos
Fator de Crescimento Epidérmico/genética , Receptores ErbB/genética , Proteínas Hedgehog/farmacologia , Receptores de Superfície Celular/genética , Glândula Submandibular/metabolismo , Animais , Western Blotting , Fator de Crescimento Epidérmico/metabolismo , Receptores ErbB/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Proteínas Hedgehog/genética , Camundongos Endogâmicos ICR , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Morfogênese/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Receptores Patched , Receptor Patched-1 , Fosforilação/efeitos dos fármacos , Gravidez , Receptor ErbB-2/genética , Receptor ErbB-2/metabolismo , Receptor ErbB-3/genética , Receptor ErbB-3/metabolismo , Receptores de Superfície Celular/metabolismo , Proteínas Recombinantes/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Glândula Submandibular/embriologiaRESUMO
The development of active water oxidation catalysts is critical to achieve high efficiency in overall water splitting. Recently, sub-10 nm-sized monodispersed partially oxidized manganese oxide nanoparticles were shown to exhibit not only superior catalytic performance for oxygen evolution, but also unique electrokinetics, as compared to their bulk counterparts. In the present work, the water-oxidizing mechanism of partially oxidized MnO nanoparticles was investigated using integrated in situ spectroscopic and electrokinetic analyses. We successfully demonstrated that, in contrast to previously reported manganese (Mn)-based catalysts, Mn(III) species are stably generated on the surface of MnO nanoparticles via a proton-coupled electron transfer pathway. Furthermore, we confirmed as to MnO nanoparticles that the one-electron oxidation step from Mn(II) to Mn(III) is no longer the rate-determining step for water oxidation and that Mn(IV)âO species are generated as reaction intermediates during catalysis.
RESUMO
By rationally designing superhydrophobic electrodes with different underwater wetting states, it is revealed that only the underwater Wenzel-Cassie coexistent state shows the clearly enhanced ability in catalyzing the oxygen reduction reaction, a typical underwater gas-consuming reaction at electrode. It is proposed that the maximizing and stabilizing the liquid/gas/solid triphase interface, endowed by the underwater Wenzel-Cassie coexistent state, plays a rather crucial role.
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Epithelial-mesenchymal interactions are required to coordinate cell proliferation, patterning, and functional differentiation of multiple cell types in a developing organ. This exquisite coordination is dependent on various secreted molecules that provide developmental signals to mediate these tissue interactions. Recently, it was reported that mature mesenchymal-derived microRNAs (miRNAs) in the fetal mouse salivary gland are loaded into exosomes, and transported to the epithelium where they influence progenitor cell proliferation. The exosomal miRNAs regulated epithelial expression of genes involved in DNA methylation in progenitor cells to influence morphogenesis. Thus, exosomal miRNAs are mobile genetic signals that cross tissue boundaries within an organ. These findings raise many questions about how miRNA signals are initiated to coordinate organogenesis and whether they are master regulators of epithelial-mesenchymal interactions. The development of therapeutic applications using exosomal miRNAs for the regeneration of damaged adult organs is a promising area of research.
Assuntos
Exossomos/metabolismo , MicroRNAs/genética , Organogênese , Transdução de Sinais , Animais , Transporte Biológico , Células Epiteliais/metabolismo , Epitélio/embriologia , Epitélio/metabolismo , Vesículas Extracelulares/genética , Vesículas Extracelulares/metabolismo , Humanos , Mesoderma/citologia , Mesoderma/embriologia , Mesoderma/metabolismo , MicroRNAs/uso terapêutico , Técnicas de Diagnóstico Molecular , Glândulas Salivares/embriologia , Glândulas Salivares/metabolismoRESUMO
The regulation of epithelial proliferation during organ morphogenesis is crucial for normal development, as dysregulation is associated with tumor formation. Non-coding microRNAs (miRNAs), such as miR-200c, are post-transcriptional regulators of genes involved in cancer. However, the role of miR-200c during normal development is unknown. We screened miRNAs expressed in the mouse developing submandibular gland (SMG) and found that miR-200c accumulates in the epithelial end buds. Using both loss- and gain-of-function, we demonstrated that miR-200c reduces epithelial proliferation during SMG morphogenesis. To identify the mechanism, we predicted miR-200c target genes and confirmed their expression during SMG development. We discovered that miR-200c targets the very low density lipoprotein receptor (Vldlr) and its ligand reelin, which unexpectedly regulate FGFR-dependent epithelial proliferation. Thus, we demonstrate that miR-200c influences FGFR-mediated epithelial proliferation during branching morphogenesis via a Vldlr-dependent mechanism. miR-200c and Vldlr may be novel targets for controlling epithelial morphogenesis during glandular repair or regeneration.
Assuntos
Células Epiteliais/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , MicroRNAs/metabolismo , Morfogênese/fisiologia , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Receptores de LDL/metabolismo , Glândula Submandibular/embriologia , Análise de Variância , Animais , Western Blotting , Proliferação de Células , Biologia Computacional , Imunofluorescência , Hibridização In Situ , Camundongos , Reação em Cadeia da Polimerase em Tempo Real , Proteína Reelina , TransfecçãoRESUMO
BACKGROUND/AIM: Oral epithelial cells serve as the primary defense against microbial exposure in the oral cavity, including the fungus Candida albicans. Dectin-1 is crucial for recognition of ß-glucan in fungi. However, expression and function of Dectin-1 in oral epithelial cells remain unclear. MATERIALS AND METHODS: We assessed Dectin-1 expression in Ca9-22 (gingiva), HSC-2 (mouth), HSC-3 (tongue), and HSC-4 (tongue) human oral epithelial cells using flow cytometry and real-time polymerase chain reaction. Cell treated with ß-glucan-rich zymosan were evaluated using real-time polymerase chain reaction. Phosphorylation of spleen-associated tyrosine kinase (SYK) was analyzed by western blotting. RESULTS: Dectin-1 was expressed in all four cell types, with high expression in Ca9-22 and HSC-2. In Ca9-22 cells, exposure to ß-glucan-rich zymosan did not alter the mRNA expression of chemokines nor of interleukin (IL)6, IL8, IL1ß, IL17A, and IL17F. Zymosan induced the expression of antimicrobial peptides ß-defensin-1 and LL-37, but not S100 calcium-binding protein A8 (S100A8) and S100A9. Furthermore, the expression of cylindromatosis (CYLD), a negative regulator of nuclear factor kappa B (NF-κB) signaling, was induced. In HSC-2 cells, zymosan induced the expression of IL17A. The expression of tumor necrosis factor alpha-induced protein 3 (TNFAIP3), a negative regulator of NF-κB signaling, was also induced. Expression of other cytokines and antimicrobial peptides remained unchanged. Zymosan induced phosphorylation of SYK in Ca9-22 cells, as well as NF-κB. CONCLUSION: Oral epithelial cells express Dectin-1 and recognize ß-glucan, which activates SYK and induces the expression of antimicrobial peptides and negative regulators of NF-κB, potentially maintaining oral homeostasis.
Assuntos
Células Epiteliais , Lectinas Tipo C , NF-kappa B , Transdução de Sinais , Quinase Syk , Humanos , Lectinas Tipo C/metabolismo , Lectinas Tipo C/genética , NF-kappa B/metabolismo , Quinase Syk/metabolismo , Quinase Syk/genética , Células Epiteliais/metabolismo , Células Epiteliais/efeitos dos fármacos , Linhagem Celular , Zimosan/farmacologia , Citocinas/metabolismo , Citocinas/genética , Fosforilação , Mucosa Bucal/metabolismo , Mucosa Bucal/imunologia , Proteínas Citotóxicas Formadoras de Poros/metabolismo , Proteínas Citotóxicas Formadoras de Poros/genética , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/metabolismoRESUMO
Strain THI201, a member of the alphaproteobacteria, is a novel thiocyanate (SCN(-))-degrading bacterium isolated from lake water enriched with potassium thiocyanate (KSCN). This bacterium carries the enzyme thiocyanate hydrolase (SCNase) that hydrolyses thiocyanate to carbonyl sulfide and ammonia. Characterization of both native and recombinant SCNase revealed properties different from known SCNases regarding subunit structure and thermostability: SCNase of strain THI201 was composed of a single protein and thermostable. We cloned and sequenced the corresponding gene and determined a protein of 457 amino acids of molecular mass 50â267 Da. Presence of a twin-arginine (Tat) signal sequence of 32 amino acids was found upstream of SCNase. The deduced amino acid sequence of SCNase showed 83% identity to that of a putative uncharacterized protein of Thiobacillus denitrificans ATCC 25259, but no significant identity to those of three subunits of SCNase from Thiobacillus thioparus strain THI115. The specific activities of native and recombinant enzyme were 0.32 and 4-15 µmol min(-1) (mg protein)(-1), respectively. The maximum activity of SCNase was found in the temperature range 30-70 °C. The thiocyanate-hydrolysing activity in both enzymes was decreased by freeze-thawing, although 25-100% of the activity of recombinant protein could be retrieved by treating the enzyme at 60 °C for 15 min. Furthermore, both native and recombinant enzymes retained the activity after pre-treatment of the protein solution at temperatures up to 70 °C.
Assuntos
Alphaproteobacteria/enzimologia , Alphaproteobacteria/genética , Hidrolases/genética , Hidrolases/metabolismo , Tiocianatos/metabolismo , Alphaproteobacteria/classificação , Alphaproteobacteria/isolamento & purificação , Sequência de Aminoácidos , Amônia/metabolismo , Sequência de Bases , Biotransformação , Clonagem Molecular , DNA Bacteriano/química , DNA Bacteriano/genética , Estabilidade Enzimática , Expressão Gênica , Hidrolases/química , Lagos/microbiologia , Dados de Sequência Molecular , Peso Molecular , Sinais Direcionadores de Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Óxidos de Enxofre/metabolismo , Temperatura , Thiobacillus/enzimologia , Thiobacillus/genéticaRESUMO
PURPOSE: In patients undergoing primary percutaneous coronary intervention (PCI) for the treatment of ST-segment elevation myocardial infarction (STEMI), coronary microvascular dysfunction is associated with poor prognosis. Coronary microvascular resistance is predominantly regulated by ATP-sensitive potassium (KATP) channels. The aim of this study was to clarify whether nicorandil, a hybrid KATP channel opener and nitric oxide donor, may be a good candidate for improving microvascular dysfunction even when administered after primary PCI. METHODS: We compared the beneficial effects of nicorandil and nitroglycerin on microvascular function in 60 consecutive patients with STEMI. After primary PCI, all patients received single intracoronary administrations of nitroglycerin (250 µg) and nicorandil (2 mg) in a randomized order; 30 received nicorandil first, while the other 30 received nitroglycerin first. Microvascular dysfunction was evaluated with the index of microcirculatory resistance (IMR), defined as the distal coronary pressure multiplied by the hyperemic mean transit time. RESULTS: As a first administration, nicorandil decreased IMR significantly more than did nitroglycerin (median [interquartile ranges]: 10.8[5.2-20.7] U vs. 2.1[1.0-6.0] U, p=0.0002).As a second administration, nicorandil further decreased IMR, while nitroglycerin did not (median [interquartile ranges]: 6.0[1.3-12.7] U vs. -1.4[-2.6 to 1.3] U, p<0.0001). The IMR after the second administration was significantly associated with myocardial blush grade, angiographic TIMI frame count after the procedure, and peak creatine kinase level. CONCLUSION: Intracoronary nicorandil reduced microvascular dysfunction after primary PCI more effectively than did nitroglycerin in patients with STEMI, probably via its KATP channel-opening effect.
Assuntos
Infarto do Miocárdio/terapia , Nicorandil/administração & dosagem , Doadores de Óxido Nítrico/administração & dosagem , Nitroglicerina/administração & dosagem , Intervenção Coronária Percutânea , Idoso , Estudos Cross-Over , Vias de Administração de Medicamentos , Feminino , Humanos , Masculino , Microvasos/efeitos dos fármacos , Microvasos/fisiopatologia , Pessoa de Meia-Idade , Infarto do Miocárdio/fisiopatologia , Resistência Vascular/efeitos dos fármacosRESUMO
Branching morphogenesis in murine submandibular glands (SMG) is regulated by growth factors, extracellular matrix (ECM) and many biological processes through interactions between the epithelium and the mesenchyme. MicroRNAs (miRNAs) are a set of small, non-protein-coding RNAs that regulate gene expression at the post-transcriptional level. We hypothesized that branching morphogenesis is partly regulated by miRNAs. Forty-four miRNAs and novel miRNA candidates were detected in SMG at embryonic day 13 by a cloning method combined with Argonaute-2 immunoprecipitation. MicroRNA21 (miR-21) expression in the mesenchyme was up-regulated and accelerated by epidermal growth factor, which is known to enhance branching morphogenesis in vitro. Down-regulation of miR-21 in the mesenchyme by locked nucleic acids was associated with a decrease in the number of epithelial buds. Relative quantification of candidates for target genes of miR-21 indicated that two messenger RNAs (for Reck and Pdcd4) were down-regulated in the mesenchyme, where miR-21 expression levels were up-regulated. These results suggest that branching morphogenesis is regulated by miR-21 through gene expression related to ECM degradation in the mesenchyme.
Assuntos
MicroRNAs/genética , MicroRNAs/metabolismo , Glândula Submandibular/embriologia , Glândula Submandibular/metabolismo , Animais , Sequência de Bases , Primers do DNA/genética , Regulação para Baixo , Fator de Crescimento Epidérmico/farmacologia , Matriz Extracelular/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Técnicas In Vitro , Mesoderma/embriologia , Mesoderma/metabolismo , Camundongos , Camundongos Endogâmicos ICR , MicroRNAs/química , Morfogênese , Conformação de Ácido Nucleico , Oligonucleotídeos/genética , Homologia de Sequência do Ácido Nucleico , Glândula Submandibular/efeitos dos fármacos , Transfecção , Regulação para CimaRESUMO
Growth factors and their receptors regulate development of many organs through activation of multiple intracellular signaling cascades including a mitogen-activated protein kinase (MAPK). Extracellular regulated kinases (ERK)1/2, classic MAPK family members, are expressed in fetal mouse submandibular glands (SMG), and stimulate branching morphogenesis. ERK5, also called big mitogen-activated protein kinase 1, was recently found as a new member of MAPK super family, and its biological roles are still largely unknown. In this study, we investigated the expression and function of ERK5 in developing fetal mouse SMGs. Western blotting analysis showed that the expression pattern of ERK5 was different from the pattern of ERK1/2 in developing fetal SMGs. Both ERK1/2 and ERK5 were phosphorylated after exposure to ligands of the ErbB family of receptor tyrosine kinases (RTKs). Phosphorylation of ERK1/2 was strongly induced by epidermal growth factor (EGF) in SMG rudiments at embryonic day 14 (E14), E16 and E18. However, ERK5 phosphorylation induced by EGF was clearly observed at E14 and E16, but not at E18. Branching morphogenesis of cultured E13 SMG rudiments was strongly suppressed by administration of U0126, an inhibitor for ERK1/2 activation, whereas the phosphorylation of ERK5 was not inhibited by U0126. BIX02188, a specific inhibitor for ERK5 activation, also inhibited branching morphogenesis in cultured SMG rudiments. These results show that EGF-responsive ERK5 is expressed in developing fetal mouse SMG, and suggest that both ERK1/2 and ERK5 signaling cascades might play an important role in the regulation of branching morphogenesis.
Assuntos
Fator de Crescimento Epidérmico/metabolismo , Feto/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Proteína Quinase 7 Ativada por Mitógeno/metabolismo , Morfogênese/fisiologia , Transdução de Sinais/fisiologia , Glândula Submandibular/embriologia , Fatores Etários , Compostos de Anilina/farmacologia , Animais , Western Blotting , Butadienos/farmacologia , Primers do DNA/genética , Feto/embriologia , Regulação da Expressão Gênica no Desenvolvimento/genética , Indóis/farmacologia , Camundongos , Proteína Quinase 7 Ativada por Mitógeno/antagonistas & inibidores , Morfogênese/efeitos dos fármacos , Nitrilas/farmacologia , Fosforilação , Receptores Proteína Tirosina Quinases/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Glândula Submandibular/metabolismoRESUMO
ATP and hydrolysis products of ATP like adenosine regulate the chemotaxis of neutrophils by activating purinoceptors and adenosine receptors. The present study was designed to examine exogenous ATP, activation of purinoceptors, and activation of A(3) adenosine receptor as key steps in the signal cascades that control cell orientation and migration of rat neutrophils. One or more of those steps might be potential therapeutic targets for treatment of inflammatory diseases. The chemotaxis of rat neutrophils was stimulated with N-formyl-methionyl-leucyl-phenylalanine (fMLP) and measured with an EZ-TAXIScan apparatus. The effects of apyrase, exogenous ATP, suramin (P2X and P2Y blocker), PPADS (a P2X blocker), TNP-ATP (P2X(1) and P2X(3) antagonist), and Reactive Blue 2 (a P2Y blocker) on the chemotactic response were also investigated. Rat neutrophil chemotaxis was significantly suppressed by apyrase. fMLP induced rat neutrophil chemotaxis was potentiated by ATP, blocked by suramin, not affected by PPADS or TNP-ATP, and significantly inhibited by RB-2. Western blotting showed that A(3), P2Y(2), and P2Y(11) were expressed in rat neutrophils. The chemotactic response of rat neutrophils to fMLP stimulation is potentiated by ATP via P2Y(11) purinoceptors but not P2X purinoceptors or A(3) adenosine receptor, and that the response plays a critical role in host defense and pathogenicity.
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Trifosfato de Adenosina/farmacologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Neutrófilos/imunologia , Receptores Purinérgicos P2Y12/fisiologia , Trifosfato de Adenosina/fisiologia , Animais , Células Cultivadas , Quimiotaxia de Leucócito/imunologia , Masculino , N-Formilmetionina Leucil-Fenilalanina/imunologia , Ratos , Ratos Wistar , Receptor A3 de Adenosina/fisiologiaRESUMO
Introduction. A diverse microbiota including fungi exists in the subgingival sites of patients with chronic periodontitis. The cell wall of Candida albicans, the most abundant fungal species, contains ß-glucan. Dectin-1 binds ß-glucan and participates in fungal recognition.Gap statement. Human periodontal ligament fibroblasts (PDLFs) are present in the periodontal ligament and synthesize immunomodulatory cytokines that influence the local response to infections. However, the expression and role of Dectin-1 in PDLFs have not been explored.Aim. This study aimed to determine if PDLFs express Dectin-1 and induce innate immune responses through Dectin-1 and the signalling molecule Syk.Methodology. The expression of Dectin-1 in PDLFs was determined by flow cytometry, western blotting and confocal microscopy. Real-time PCR and Western blotting were used to determine the immune response of PDLFs stimulated with ß-glucan-rich zymosan and C. albicans.Results. Dectin-1 was constitutively expressed in PDLFs. Zymosan induced the expression of cytokines, including IL6, IL1B and IL17A, and the chemokine IL8. Zymosan also induced the expression of the antimicrobial peptide ß-defensin-1 (DEFB1). Further, the phosphorylation of Syk and NF-κB occurred upon Dectin-1 activation. Notably, heat-killed C. albicans induced the expression of IL6, IL17A, IL8 and DEFB1, and this activation was suppressed by the Syk inhibitor, R406.Conclusion. These findings indicate that the Dectin-1/Syk pathway induces an innate immune response of PDLFs, which may facilitate the control of oral infections such as candidiasis and periodontitis.
Assuntos
Fibroblastos , Ligamento Periodontal , Quinase Syk , beta-Defensinas , Humanos , beta-Defensinas/imunologia , Candida albicans/metabolismo , Citocinas , Fibroblastos/imunologia , Imunidade Inata , Interleucina-6 , Interleucina-8 , Ligamento Periodontal/citologia , Ligamento Periodontal/imunologia , Quinase Syk/imunologia , ZimosanRESUMO
BACKGROUND: XE991 (10,10-bis(4-pyridinylmethyl)-9(10H)-anthracenone) is reportedly a potent and selective Kv7 (KCNQ) channel inhibitor. This study aimed to evaluate how XE991 affects nicotinic responses in intracardiac ganglion neurons. METHODS: We studied how the KCNQ channel inhibitor XE991 could affect nicotinic responses in acutely isolated rat intracardiac ganglion neurons using a perforated patch-clamp recording configuration and Ca2+ imaging. RESULTS: XE991 reversibly and concentration-dependently inhibited the nicotine (10 µM)-induced current with an IC50 of 14.4 µM. The EC50 values for nicotine-induced currents in the absence and presence of 10 µM XE991 were 8.7 and 12.0 µM, respectively. Because XE991 suppressed the maximum response of the nicotine concentration-response curve, the inhibitory effect of this drug appears to be noncompetitive. In addition, linopirdine reduced the amplitude of 10 µM nicotine-induced currents with an IC50 value of 16.9 µM. The inorganic KCNQ channel inhibitor Ba2+ affected neither the nicotine-induced current nor the inhibitory effect of XE991 on the nicotinic response. The KCNQ activator flupirtine at a concentration of 10 µM slightly but markedly inhibited the nicotine-induced current. Finally, XE991 inhibited the nicotine-induced elevation of intracellular calcium concentration and the nicotine-induced firing of action potentials. CONCLUSION: We propose that XE991 inhibits nicotinic acetylcholine receptors in intracardiac ganglion neurons, which in turn attenuate nicotine-induced neuronal excitation.
Assuntos
Receptores Nicotínicos , Animais , Neurônios , Nicotina/farmacologia , RatosRESUMO
BACKGROUND: The objective of this study was to investigate whether a distal protection (DP) device can preserve the index of microcirculatory resistance (IMR) after primary percutaneous coronary intervention (PCI) in patients with anterior ST-segment elevation myocardial infarction (STEMI). METHODS AND RESULTS: The study group of 36 consecutive patients with anterior STEMI were randomized into 2 groups of primary PCI with or without DP: stenting without DP (non-DP group, n = 17) and with DP (DP group, n = 19). The DP in all cases was Filtrap (Nipro, Japan). Following final coronary angiography after successful PCI, IMR was measured using PressureWire™ Certus (St Jude Medical, USA) at maximal hyperemia. The averaged IMR of the 36 patients with STEMI after primary PCI was 31.6U. The IMR in the DP group was significantly lower than that in the non-DP group (26.6 ± 25.8U vs. 37.2 ± 23.2U, P = 0.03242). CONCLUSIONS: DP as an adjunctive therapy of PCI for acute anterior STEMI may have beneficial effects on myocardial microcirculation because of preservation of IMR.
Assuntos
Angioplastia Coronária com Balão/instrumentação , Infarto Miocárdico de Parede Anterior/terapia , Circulação Coronária , Dispositivos de Proteção Embólica , Microcirculação , Stents , Resistência Vascular , Angioplastia Coronária com Balão/efeitos adversos , Angioplastia Coronária com Balão/mortalidade , Infarto Miocárdico de Parede Anterior/diagnóstico por imagem , Infarto Miocárdico de Parede Anterior/mortalidade , Infarto Miocárdico de Parede Anterior/fisiopatologia , Angiografia Coronária , Humanos , Japão , Estudos Prospectivos , Fatores de Tempo , Resultado do TratamentoRESUMO
[This corrects the article DOI: 10.1016/j.jdsr.2017.06.002.].
RESUMO
BACKGROUND: Although microvascular dysfunction following percutaneous coronary intervention (PCI) can be evaluated with the index of microcirculatory resistance (IMR), no method of treatment has been established. We hypothesized that intracoronary administration of nicorandil can improve IMR after successful primary PCI in patients with ST-segment elevation myocardial infarction (STEMI). METHODS AND RESULTS: In 40 patients with first STEMI after successful primary PCI, IMR was measured using PressureWire(TM) Certus (St. Jude Medical, MN, USA). In 20 of the patients (Group N), IMR was measured at baseline and after intracoronary nicorandil (2 mg/10 ml). In the other 20 patients (Control), IMR was measured at baseline, after intracoronary saline (10 ml) and after intracoronary nicorandil (2 mg/10 ml). In Group N, IMR significantly decreased after intracoronary nicorandil (median IMR, 27.7-18.7 U, P<0.0001). In the Control group, IMR did not change after saline administration (median IMR, 24.3-23.8 U, P=0.8193), but was significantly decreased after intracoronary nicorandil (median IMR, 23.8-14.9 U, P<0.0001). Next, all 40 patients were divided into subgroups by tertile of baseline IMR. In those with intermediate to high IMR (baseline IMR > or =21), intracoronary nicorandil significantly decreased IMR, although it did not change IMR in those with low IMR (baseline IMR <21). CONCLUSIONS: High IMR levels in patients with STEMI after successful primary PCI can be improved by intracoronary administration of nicorandil.