RESUMEN
A novel, practical protocol for the aerobic direct C-H acetoxylation of arenes, employing a recyclable heterogeneous rhodium catalyst, is reported herein. The trifluoroacetoxylation of 2-amido-substituted anthracenes proceeded at the 9-position with exclusive regioselectivity. The oxidation of variously substituted anthracenes and other polycyclic aromatics with molecular oxygen as a terminal oxidant proceeded under mild conditions, providing products in good to excellent yields.
RESUMEN
BACKGROUND: The activation of liver X receptor (LXR) α or LXRß negatively regulates the expression of pro-inflammatory genes in mammalian cells. We recently reported that 25-hydroxycholesterol, a representative LXR-activating oxysterol, suppresses IL-6 production in mouse mast cells (MCs) following its engagement of the high-affinity IgE receptor (FcεRI). This finding suggests that murine MCs express functional LXRs; however, the mechanisms underlying the LXR-dependent repression of the MC-mediated production of pro-inflammatory cytokines, including IL-6, are poorly understood. Therefore, we employed the synthetic LXR ligand GW3965 to examine the functions of LXRα and LXRß in the production of pro-inflammatory cytokines by murine bone marrow-derived MCs (BMMCs). METHODS: We prepared BMMCs from wild-type (WT), LXRα(-/-), and LXRα/ß(-/-) mice. Each group of BMMCs was pretreated with GW3965 and then stimulated with IgE+antigen (Ag) or lipopolysaccharide (LPS). Cytokine production was then analyzed using specific ELISA kits. RESULTS: The activation of LXRs by GW3965 significantly attenuated the production of IL-1α and IL-1ß, but not of IL-6, in the WT and LXRα(-/-) BMMCs stimulated with IgE+Ag. However, GW3965 treatment decreased the production of IL-1α, IL-1ß, and IL-6 in WT and LXRα(-/-) BMMCs upon stimulation with LPS, while the GW3965-mediated suppression of cytokine production was nearly absent from the LXRα/ß(-/-) BMMCs. CONCLUSIONS: These findings demonstrate, for the first time, that the activation of LXRs by GW3965 attenuates the antigen- or LPS-induced production of pro-inflammatory cytokines, such as IL-1α and IL-1ß, in murine MCs and that LXRß plays an important role in the LXR-mediated repression of cytokine production.
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Benzoatos/farmacología , Bencilaminas/farmacología , Citocinas/metabolismo , Mediadores de Inflamación/metabolismo , Mastocitos/efectos de los fármacos , Mastocitos/fisiología , Receptores Nucleares Huérfanos/agonistas , Animales , Degranulación de la Célula/genética , Degranulación de la Célula/inmunología , Citocinas/genética , Expresión Génica , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Lipopolisacáridos/inmunología , Receptores X del Hígado , Ratones , Ratones Noqueados , Receptores Nucleares Huérfanos/genética , Receptores Nucleares Huérfanos/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de IgE/agonistas , Receptores de IgE/metabolismoAsunto(s)
Anticuerpos Monoclonales Humanizados/efectos adversos , Antirreumáticos/administración & dosificación , Artritis Reumatoide/tratamiento farmacológico , Fascitis Necrotizante/inducido químicamente , Infecciones Estreptocócicas/inducido químicamente , Infección de Heridas/inducido químicamente , Antibacterianos/uso terapéutico , Artritis Reumatoide/diagnóstico , Artritis Reumatoide/inmunología , Biopsia , Terapia Combinada , Desbridamiento , Fascitis Necrotizante/diagnóstico , Fascitis Necrotizante/microbiología , Fascitis Necrotizante/terapia , Humanos , Masculino , Persona de Mediana Edad , Trasplante de Piel , Infecciones Estreptocócicas/diagnóstico , Infecciones Estreptocócicas/microbiología , Infecciones Estreptocócicas/terapia , Resultado del Tratamiento , Cicatrización de Heridas , Infección de Heridas/diagnóstico , Infección de Heridas/microbiología , Infección de Heridas/terapiaRESUMEN
OBJECTIVE: Substantial evidence suggests that human synovial mast cells (MCs) are involved in the pathogenesis of rheumatoid arthritis (RA). A plausible pathway for the activation of synovial MCs is through IgG receptors, given the prevalence of circulating IgG isotype autoantibodies and synovial immune complexes in patients with RA. However, IgG receptor expression on human synovial MCs remains uncharacterized. The aim of this study was to identify which IgG receptor(s) on synovial MCs are responsible for MC activation in immune complexes. METHODS: Synovial tissue specimens were obtained from patients with RA or patients with osteoarthritis (OA) who were undergoing joint replacement surgery, and synovial MCs were enzymatically dispersed. Cultured synovium-derived MCs were generated by culturing synovial cells with stem cell factor, and receptor expression was analyzed using fluorescence-activated cell sorting. Mediators released from MCs were measured using enzyme immunoassays or enzyme-linked immunosorbent assays. RESULTS: Primary synovial MCs and cultured synovium-derived MCs obtained from both patients with RA and patients with OA expressed Fcε receptor I (FcεRI), FcγRI, and FcγRII but not FcγRIII. Cultured synovium-derived MCs induced degranulation and the production of prostaglandin D2 and tumor necrosis factor α (TNFα) through FcγRI. The aggregation of FcγRII caused histamine release from cultured MCs but not from primary MCs. Histamine release induced by aggregated IgG was significantly inhibited by neutralizing anti-FcγRI monoclonal antibody and anti-FcγRII monoclonal antibody. CONCLUSION: With regard to the FcR expression profile, synovial MCs from patients with RA and patients with OA were similar. FcγRI was responsible for producing abundant TNFα from synovial MCs in response to aggregated IgG. Immune complexes may activate synovial MCs through FcγRI and FcγRII.
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Artritis Reumatoide/inmunología , Inmunoglobulina G/inmunología , Mastocitos/inmunología , Osteoartritis/inmunología , Receptores de IgG/inmunología , Membrana Sinovial/inmunología , Artritis Reumatoide/metabolismo , Artritis Reumatoide/patología , Células Cultivadas , Citometría de Flujo , Humanos , Inmunohistoquímica , Técnicas In Vitro , Mastocitos/metabolismo , Osteoartritis/metabolismo , Osteoartritis/patología , Receptores de IgG/metabolismo , Membrana Sinovial/metabolismoRESUMEN
BACKGROUND: A large body of evidence has demonstrated that treatment with omalizumab is clinically effective for the management of moderate to severe allergic asthma, emphasizing the importance of IgE in the pathogenesis of allergic asthma. We hypothesized that IgE accelerates FcεRI-mediated responsiveness of "immature" human mast cells (MCs) and that omalizumab downregulates the acceleration. OBJECTIVES: To examine when MC progenitors acquired the ability to degranulate following FcεRI aggregation, whether IgE accelerates the responsiveness of immature MCs following FcεRI aggregation, and whether omalizumab regulates such an acceleration. METHODS: Gene expression was examined using a microarray and quantitative reverse transcription polymerase chain reaction. Protein expression was investigated using FACS. Histamine release was examined using an EIA. RESULTS: The time-course analysis of the mRNA expression of MC-related genes, including FcεRI, in Kit(+) sorted cells during the differentiation and histamine experiments revealed that the expression level of FcεRI in 5 week (w)-cultured MCs was not sufficient to induce degranulation following FcεRI aggregation but that 5 w-cultured MCs were fully responsive to calcium ionophore. By addition of IgE in culture medium FcεRI expression level and FcεRI-mediated histamine release of 5 w-cultured MCs were significantly increased compared with those without addition of IgE, whereas the expression level of tryptase and number of MCs was not affected. Omalizumab significantly inhibited IgE-dependent enhancement of FcεRI expression level and FcεRI-mediated histamine release. CONCLUSIONS: High levels of IgE in the microenvironment in vivo may upregulate the responsiveness of immature MCs to allergens. Omalizumab may inhibit the IgE-mediated responsiveness of not only mature MCs, but also immature MCs.
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Anticuerpos Antiidiotipos/farmacología , Anticuerpos Monoclonales Humanizados/farmacología , Inmunoglobulina E/inmunología , Mastocitos/inmunología , Receptores de IgE/inmunología , Asma/patología , Células Cultivadas , Expresión Génica , Liberación de Histamina , Humanos , Inmunoglobulina E/metabolismo , Mastocitos/metabolismo , Omalizumab , ARN Mensajero/biosíntesis , Receptores de IgE/biosíntesisRESUMEN
BACKGROUND: CCL23 (MPIF1/CK-BETA-8) is a novel CC chemokine that plays important roles in the inhibition of myeloid progenitor cell development, the selective recruitment of resting T lymphocytes and monocytes, and the potentiation of VEGF-induced proliferation and migration of human endothelial cells. Since eosinophils participate in the pathogenesis of airway remodeling, we examined CCL23 production and release by human eosinophils in vitro. METHODS: Using Ficoll and antibody-coated immunomagnetic beads, eosinophils and other blood cells were purified from peripheral blood samples obtained from normal subjects and mildly allergic patients. Eosinophils were cultured in the presence of 10 ng/ml granulocyte-macrophage colony-stimulating factor (GM-CSF), 10 ng/ml IL-5, 100 ng/ml IFN-γ, 100 ng/ml IFN-α, or immobilized secretory IgA (sIgA). Total mRNA was extracted after 6 h of culture, and mRNA expression was measured using a microarray and RT-PCR. The CCL23 concentrations in the supernatants and cell lysates after 24 and 48 h of culture were measured by ELISA. RESULTS: CCL23 mRNAs (both CK-ß8-1 and CK-ß8) were constitutively expressed in fresh eosinophils, and their expression levels were higher than in other types of blood cells. CCL23 mRNAs were significantly increased by stimulation with GM-CSF and IL-5 and slightly by IFN-α and immobilized sIgA. Fresh eosinophils contained trace amounts of CCL23 protein. CCL23 was significantly released into the supernatant when the eosinophils were stimulated with GM-CSF or IL-5 but not with IFN-γ or immobilized sIgA. CONCLUSION: Our data suggest that eosinophils produce and release CCL23 and may be involved in some in vivo physiological and pathological conditions.
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Quimiocinas CC/metabolismo , Eosinófilos/metabolismo , Expresión Génica/inmunología , Células Sanguíneas/metabolismo , Quimiocinas CC/genética , Medios de Cultivo Condicionados/metabolismo , Eosinófilos/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Humanos , Inmunoglobulina A Secretora/farmacología , Interferón-alfa/farmacología , Interferón gamma/farmacología , Interleucina-5/farmacología , Análisis de Secuencia por Matrices de Oligonucleótidos , Isoformas de Proteínas/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Crecimiento Transformador beta/farmacologíaRESUMEN
BACKGROUND: In recent years, it has been reported that stem cells exist in the mesenchymal tissues of the bone marrow and adipose. These stem cells are thought to express specific cell surface markers such as CD44, CD54, CD105, CD90, and CD271 and have been confirmed to be pluripotent. Furthermore, although it has been reported that stem cells are also present in the dermis, their cell surface markers and characteristics are not fully understood. OBJECTIVE: To confirm the presence of stem cells in the dermis and their ability, employing the mesenchymal stem cell markers which have previously been reported as an indication. METHODS: We analyzed the percentages of CD44 (+), CD54 (+), CD90 (+), CD105 (+), and CD271 (+) cells in the dermis of neonatal mice (HR-1 mouse) by performing immunostaining and FACS. Secondly, we isolated each type of marker-positive and -negative cells from dermal tissues and evaluated their proliferation potential and their ability to differentiate into adipocytes, osteoblasts, and chondrocytes. RESULTS: According to the immunostaining and FACS results, we confirmed that stem cells that express CD44, CD54, CD90, CD105, and CD271 are present in the dermal tissues of neonatal mice. In addition, when we measured the proliferation and differentiation potentials of each type of marker-positive cells, it was revealed that cells expressing CD54 or CD271 have a high proliferation potential and are able to differentiate into adipocytes, osteoblasts, and chondrocytes. CONCLUSIONS: These results indicated that dermal tissues contain stem cells that express CD44, CD54, CD90, CD105, and CD271 which are stem cell markers. More precisely, it was suggested that both CD54 (+) and CD271 (+) stem cells have high proliferation and differentiation potentials.
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Membrana Celular/metabolismo , Dermis/metabolismo , Regulación de la Expresión Génica , Adapaleno , Adipocitos/citología , Animales , Proliferación Celular , Condrocitos/citología , Dermatología/métodos , Endoglina , Citometría de Flujo , Receptores de Hialuranos/biosíntesis , Molécula 1 de Adhesión Intercelular/biosíntesis , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Ratones , Naftalenos/metabolismo , Osteoblastos/citología , Células Madre/citología , Células Madre/metabolismo , Antígenos Thy-1/biosíntesisRESUMEN
BACKGROUND: Respiratory syncytial virus (RSV) infection in infants with Th2 predisposition is thought to increase the risk of allergic sensitization, recurrent wheezing, and bronchial asthma during childhood. We attempted to clarify the molecular mechanisms by which Th1/Th2 predisposition in the host alters RSV infection and facilitates airway inflammation. METHODS: A549 human airway epithelial cells were inoculated with live or UV-treated RSV after pretreatment with either a combination of tumor necrosis factor (TNF)-α and interferon-γ (Th1-primed) or a combination of TNF-α and interleukin-4 (Th2-primed) for 48 h. The gene and protein expression profiles of RSV-infected A549 cells were examined. RESULTS: GeneChip analysis indicated that, at 96 h after inoculation with RSV, the expression of 62 genes was specifically enhanced (more than 2-fold by normalized data) in Th2-primed cells compared to that in unprimed or Th1-primed cells. An increase in mRNA and protein levels of monocyte chemoattractant protein (MCP)-1/CCL2 among those 62 genes was confirmed by real-time PCR and cytometric bead assay, respectively. RSV replication was markedly diminished in Th1-primed airway epithelial cells but not in Th2-primed cells, which was presumably caused at least in part by the early induction of antiviral genes. CONCLUSIONS: These results suggest that Th1/Th2 predisposition in the host prior to RSV infection critically regulates inflammatory reactions in the airways through alteration of gene expression, and that MCP-1/CCL2 plays an important role in the pathogenesis of severe RSV infection and the subsequent development of asthma in Th2-predisposed hosts.
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Quimiocina CCL2/metabolismo , Células Epiteliales/metabolismo , Regulación de la Expresión Génica , Interferón gamma/farmacología , Pulmón/metabolismo , Virus Sincitial Respiratorio Humano/patogenicidad , Factor de Necrosis Tumoral alfa/farmacología , Línea Celular , Quimiocina CCL2/genética , Citocinas/genética , Citocinas/metabolismo , Células Epiteliales/efectos de los fármacos , Células Epiteliales/inmunología , Células Epiteliales/virología , Perfilación de la Expresión Génica , Humanos , Interferón gamma/metabolismo , Pulmón/citología , Pulmón/efectos de los fármacos , Pulmón/inmunología , Pulmón/virología , Análisis de Secuencia por Matrices de Oligonucleótidos , Virus Sincitial Respiratorio Humano/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células TH1/inmunología , Células TH1/metabolismo , Células Th2/inmunología , Células Th2/metabolismo , Factores de Tiempo , Factor de Necrosis Tumoral alfa/metabolismo , Replicación ViralRESUMEN
IL-33 is a member of the IL-1 family and mediates its biological effects via the ST2 receptor, which is selectively expressed on Th2 cells and mast cells. Although polymorphic variation in ST2 is strongly associated with asthma, it is currently unclear whether IL-33 acts directly on lung tissue cells at sites of airway remodeling. Therefore, we aimed to identify the IL-33-responsive cells among primary human lung tissue cells. ST2 mRNA was expressed in both endothelial and epithelial cells but not in fibroblasts or smooth muscle cells. Correspondingly, IL-33 promoted IL-8 production by both endothelial and epithelial cells but not by fibroblasts or smooth muscle cells. Transfection of ST2 small interference RNA into both endothelial and epithelial cells significantly reduced the IL-33-dependent upregulation of IL-8, suggesting that IL-33-mediated responses in these cells occur via the ST2 receptor. Importantly, Th2 cytokines, such as IL-4, further enhanced ST2 expression and function in both endothelial and epithelial cells. The IL-33-mediated production of IL-8 by epithelial cells was almost completely suppressed by corticosteroid treatment. In contrast, the effect of corticosteroid treatment on the IL-33-mediated responses of endothelial cells was only partial. IL-33 induced activation of both ERK and p38 MAPK in endothelial cells but only ERK in epithelial cells. p38 MAPK was required for the IL-33-mediated responses of endothelial cells, whereas ERK was required for IL-33-mediated IL-8 production by epithelial cells. Taken together, these findings suggest that IL-33-mediated inflammatory responses of lung tissue cells may be involved in the chronic allergic inflammation of the asthmatic airway.
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Inflamación/inmunología , Interleucinas/inmunología , Pulmón/inmunología , Transducción de Señal/inmunología , Asma/metabolismo , Western Blotting , Células Cultivadas , Células Endoteliales/inmunología , Células Endoteliales/metabolismo , Ensayo de Inmunoadsorción Enzimática , Células Epiteliales/inmunología , Células Epiteliales/metabolismo , Fibroblastos/inmunología , Fibroblastos/metabolismo , Humanos , Inflamación/metabolismo , Proteína 1 Similar al Receptor de Interleucina-1 , Interleucina-33 , Interleucina-8/inmunología , Interleucina-8/metabolismo , Interleucinas/metabolismo , Pulmón/citología , Pulmón/metabolismo , Miocitos del Músculo Liso/inmunología , Miocitos del Músculo Liso/metabolismo , ARN Interferente Pequeño , Receptores de Superficie Celular/inmunología , Receptores de Superficie Celular/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , TransfecciónRESUMEN
Lens epithelium-derived growth factor (LEDGF)/dense fine speckles 70 kDa protein (DFS70) is a transcription cofactor that enhances growth and is overexpressed in various cancers. In the epidermis, LEDGF/DFS70 localizes to the nucleus of keratinocytes (KCs) in the basal layers and to the cytoplasm of cells in the upper layers. However, the biological and pathological relevance of LEDGF/DFS70 in the epidermis is virtually unknown. Compared with normal epidermis, we detected strong nuclear staining of LEDGF/DFS70 in both the spinous and basal layers of the epidermis of psoriatic skin. To investigate the roles of LEDGF/DFS70 in the epidermis of psoriatic skin, we generated HaCaT cells that constitutively express enhanced green fluorescence protein (EGFP)-LEDGF (EGFP-LEDGF-HaCaT) or EGFP alone (EGFP-HaCaT) as a control. EGFP-LEDGF-HaCaT cells had increased expression of IL-6, which was attenuated by LEDGF-specific RNA interference and the p38-specific inhibitors SB-239063 and SB-203580. Furthermore, EGFP-LEDGF-HaCaT cells had increased expression of S100A7 and S100A9 and decreased expression of filaggrin. These findings are compatible with the expression pattern in psoriatic tissues. Taken together, these results strongly suggest that ectopic expression of LEDGF/DFS70 in KCs could be involved in the pathology of psoriasis vulgaris.
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Proteínas Adaptadoras Transductoras de Señales/genética , Interleucina-6/genética , Queratinocitos/fisiología , Psoriasis , Factores de Transcripción/genética , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Adolescente , Adulto , Anciano , Calgranulina B/genética , Carcinoma de Células Escamosas , Epidermis/metabolismo , Epidermis/patología , Epidermis/fisiopatología , Proteínas Filagrina , Proteínas Fluorescentes Verdes/genética , Células HeLa , Humanos , Interleucina-6/metabolismo , Proteínas de Filamentos Intermediarios/genética , Queratinocitos/citología , Persona de Mediana Edad , Fosforilación/fisiología , Psoriasis/metabolismo , Psoriasis/patología , Psoriasis/fisiopatología , ARN Interferente Pequeño , Proteína A7 de Unión a Calcio de la Familia S100 , Proteínas S100/genética , Factor de Transcripción STAT3/metabolismo , Neoplasias Cutáneas , Factores de Transcripción/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/genéticaRESUMEN
BACKGROUND: Whilebeta(2)-adrenoceptor agonists (beta(2)-agonists) are widely used as bronchodilators in the treatment of asthma, there has been increasing concern that regular use of beta(2)-agonists may adversely affect the control of asthma. However, the molecular mechanisms of such undesirable effects of beta(2)-agonists are not fully understood. In this study, we examined the effects of beta(2)-agonists on cytokine-induced production of thymic stromal lymphopoietin (TSLP), an indispensable cytokine in the development of allergic diseases, by lung tissue cells. METHODS: Normal human bronchial epithelial cells (NHBE), smooth muscle cells (BSMC) and fibroblasts (NHLF) were stimulated with the IL-4 and TNF-alpha cytokines, alone and in combination, and their production of TSLP was examined by ELISA. The effects of beta(2)-agonists (salmeterol, formoterol, salbutamol), intracellular cyclic adenosine monophosphate (cAMP)-elevating agents (8-bromo-cAMP, dibutyryl cAMP, forskolin) and a corticosteroid (fluticasone) on the cytokine-induced TSLP production were examined. RESULTS: The following results were observed in all three types of lung tissue cells tested (that is, NHBE, BSMC and NHLF). Costimulation with IL-4 and TNF-alpha significantly induced TSLP production, and beta(2)-agonists further enhanced it via upregulation of intracellular cAMP. However, addition of a corticosteroid to the cytokines and beta(2)-agonist resulted in a marked decrease in TSLP production. CONCLUSIONS: beta(2)-Agonists significantly enhanced the cytokine-induced TSLP production by primary human lung tissue cells. This may be partly responsible for the undesirable clinical effects of continuous beta(2)-agonist monotherapy, and combination therapy with a corticosteroid might effectively inhibit TSLP-mediated allergic inflammation.
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Agonistas Adrenérgicos beta/farmacología , Broncodilatadores/farmacología , Fibroblastos/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Mucosa Respiratoria/efectos de los fármacos , Androstadienos/farmacología , Asma/tratamiento farmacológico , Células Cultivadas , AMP Cíclico/metabolismo , Citocinas/biosíntesis , Citocinas/genética , Quimioterapia Combinada , Fibroblastos/metabolismo , Fibroblastos/patología , Fluticasona , Humanos , Interleucina-4/metabolismo , Pulmón/metabolismo , Pulmón/patología , Miocitos del Músculo Liso/metabolismo , Miocitos del Músculo Liso/patología , Mucosa Respiratoria/metabolismo , Mucosa Respiratoria/patología , Factor de Necrosis Tumoral alfa/metabolismo , Linfopoyetina del Estroma TímicoRESUMEN
The unfolded protein response (UPR), which is induced by stress to the endoplasmic reticulum (ER), is involved in the functional alteration of certain cells, such as the differentiation of B cells to plasma cells. The aim of this study is to determine whether the UPR is activated during epidermal keratinocyte (KC) differentiation. Here, we show that the expression of the UPR-induced proteins Bip/GRP78 and HRD1 was increased in cells in the supra-basal layers of normal human epidermis that contain KCs undergoing differentiation as well as in skin-equivalent cultured KCs. However, Bip/GRP78 and HRD1 were poorly expressed in proliferating KCs in squamous cell carcinoma and psoriasis vulgaris tissues. The epidermal growth factor receptor tyrosine kinase inhibitor, PD153035, which induces KC differentiation, upregulated UPR-induced marker mRNAs and proteins. Furthermore, microarray analyses and quantitative PCR revealed that ER stress-inducing reagents, tunicamycin (TU), thapsigargin, and brefeldin A, altered the expression of genes essential for human epidermal KC differentiation, including C/EBPbeta, KLF4, and ABCA12 in vitro. However, ABCA12 and KLF4 mRNA did not increase with TU treatment after siRNA-mediated knockdown of XBP-1. Taken together, our findings strongly suggest that the UPR is activated during normal epidermal KC differentiation and induces C/EBPbeta, KLF4, and ABCA12 mRNAs.
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Retículo Endoplásmico/metabolismo , Queratinocitos/metabolismo , Pliegue de Proteína , Transducción de Señal/fisiología , Transportadoras de Casetes de Unión a ATP/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Proteína beta Potenciadora de Unión a CCAAT/genética , Carcinoma de Células Escamosas/química , Diferenciación Celular , Proteínas de Unión al ADN/genética , Chaperón BiP del Retículo Endoplásmico , Proteínas de Choque Térmico/análisis , Proteínas de Choque Térmico/genética , Humanos , Inmunohistoquímica , Queratinocitos/citología , Factor 4 Similar a Kruppel , Factores de Transcripción de Tipo Kruppel/genética , Persona de Mediana Edad , Proteína Disulfuro Isomerasas/análisis , Proteína Disulfuro Isomerasas/genética , Transporte de Proteínas , Psoriasis/metabolismo , Quinazolinas/farmacología , ARN Mensajero/análisis , Factores de Transcripción del Factor Regulador X , Factores de Transcripción/genética , Ubiquitina-Proteína Ligasas/análisis , Proteína 1 de Unión a la X-BoxRESUMEN
The track records of the use of anti-methicillin-resistant Staphylococcus aureus agents (anti-MRSA agents) in a 5-year period (2001.4-2006.3) were collected, and cases in which anti-MRSA agents were used for >4 days were selected. In each case, the results of laboratory data and bacterial examination before and after administering the anti-MRSA agents were investigated retrospectively. In addition, it was also investigated in each case whether therapeutic drug monitoring (TDM) was carried out. It was observed that the number of patients treated with anti-MRSA agents and the total dose of anti-MRSA agents used tended to increase over time, except for arbekacin sulfate. It was, however, shown that treatment with anti-MRSA agents resulted in significant decreases in body temperature, C-reactive protein, and white blood cell counts. Bacterial examination was conducted in 75.6% of the patients treated with anti-MRSA agents, with MRSA being detected in 72.4% of the cases examined. On the other hand, TDM was also conducted in 60% of the cases, but this was at a lower percentage than that of the other examinations. Quantitative bacterial examination after treatment with anti-MRSA agents indicates that TDM can be considered important for the appropriate use of anti-MRSA agents.
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Antibacterianos/administración & dosificación , Dibekacina/análogos & derivados , Monitoreo de Drogas , Infecciones Estafilocócicas/tratamiento farmacológico , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/efectos de los fármacos , Teicoplanina/administración & dosificación , Vancomicina/administración & dosificación , Antibacterianos/farmacología , Dibekacina/administración & dosificación , Dibekacina/farmacología , Farmacorresistencia Bacteriana , Humanos , Resistencia a la Meticilina , Estudios Retrospectivos , Staphylococcus aureus/aislamiento & purificación , Teicoplanina/farmacología , Factores de Tiempo , Vancomicina/farmacologíaRESUMEN
Follicle-associated epithelium (FAE) covering Peyer's patches contains specialized epithelial M cells that take up ingested macromolecules and microorganisms from the lumen of the gut by transcytosis. Using high-density oligonucleotide microarrays, we analyzed the gene expression profiles of FAE and M cells in order to characterize their cellular phenotypes. The microarray data revealed that, among approximately 14,000 genes, 409 were expressed in FAE at twofold or higher levels compared to the intestinal epithelial cells (IECs) of the villi. These included genes involved in membrane traffic, host defense and transcriptional regulation, as well as uncharacterized genes. Real-time PCR and in situ hybridization analyses identified three molecules, ubiquitin D (Ub-D), tumor necrosis factor receptor superfamily 12a (TNFRsf12a), and transmembrane 4 superfamily 4 (Tm4sf4), which were predominantly distributed throughout FAE, but were expressed little, if at all, in IECs. By contrast, transcripts of secretory granule neuroendocrine protein 1 (Sgne-1) were scattered in FAE, and were co-localized with Ulex europaeus agglutinin-1 (UEA-1)-positive cells. This clearly suggests that expression of Sgne-1 in the gut is specific to M cells. Such a unique pattern of gene expression distinguishes FAE and M cells from IECs, and may reflect their cellular phenotype(s) associated with specific functional features.
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Células Epiteliales/metabolismo , Ganglios Linfáticos Agregados/metabolismo , Proteoma/metabolismo , Animales , Perfilación de la Expresión Génica , Mucosa Intestinal/metabolismo , Linfocitos/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Proteína 7B2 Secretora Neuroendocrina/biosíntesis , Análisis de Secuencia por Matrices de OligonucleótidosRESUMEN
Respiratory epithelial cells play important roles not only in host defense mechanisms, but also in inflammatory responses. Inhaled corticosteroids are widely used for the treatment of patients with inflammatory lung disorders, including asthma, chronic obstructive pulmonary disease, and sarcoidosis. Corticosteroids effectively reduce the production of inflammatory mediators, such as cytokines and chemokines. Although these molecules are also essential for host defense responses, there is no convincing evidence that inhaled corticosteroids increase susceptibility to lower respiratory tract infections. To test the involvement of Toll-like receptor (TLR) family molecules in this phenomenon, we examined the effects of various cytokines and corticosteroid on the expression of TLRs in human respiratory epithelial cells. Among the TLRs tested, TLR2 expression was significantly enhanced after stimulation with a combination of tumor necrosis factor-alpha and interferon-gamma. Dexamethasone synergistically enhanced TLR2 expression in combination with tumor necrosis factor-alpha and interferon-gamma in terms of both mRNA and protein levels. Furthermore, increased cell-surface TLR2 was functional, judging from the remarkable induction of interleukin-6, interleukin-8, and beta-defensin-2 after stimulation with peptidoglycan. These results provide evidence for a novel function of corticosteroids in airway inflammatory disorders, and indicate that the use of inhaled corticosteroids in such disorders may have a beneficial role in host defense mechanisms.
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Bronquios/metabolismo , Sinergismo Farmacológico , Células Epiteliales/metabolismo , Glicoproteínas de Membrana/metabolismo , Receptores de Superficie Celular/metabolismo , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Bronquios/citología , Bronquios/inmunología , Dexametasona/administración & dosificación , Células Epiteliales/citología , Células Epiteliales/inmunología , Regulación de la Expresión Génica , Humanos , Interferón gamma/administración & dosificación , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Peptidoglicano/farmacología , Receptor Toll-Like 2 , Receptores Toll-Like , Células Tumorales Cultivadas , Factor de Necrosis Tumoral alfa/administración & dosificación , beta-Defensinas/metabolismoRESUMEN
Esophageal squamous cell carcinoma has heterogeneous clinical outcomes that cannot be predicted well using any existing clinical or molecular prognostic factors. Gene expression profiling may enable more precise prediction of the clinical outcome of these patients. We developed a new approach using gene expression ratios of paired cancerous and normal tissue specimens from the same patient to reduce the effects of variation among individuals. Using oligonucleotide microarrays, we analyzed total RNA expression levels corresponding to 12,600 transcript sequences in 24 paired cancerous and normal tissue operative specimens from 12 patients with esophageal squamous cell carcinoma. Hierarchical clustering analysis using gene expression ratios (cancer:normal) divided the 12 patients into two groups; all 7 patients in the first cluster survived without relapse (median follow-up, 483 days), whereas all 5 patients in the second cluster relapsed (median relapse-free survival time, 280 days; log-rank test, P = 0.006). In contrast, clustering either with cancerous tissue alone or with normal tissue alone did not show significant differences in the outcomes. The expressions of a variety of genes related with cell cycle, gene-repair, apoptosis and chemoradiotherpay resistance were up-regulated in the poor prognostic cluster. These results suggest that ratios of paired gene expression profiles may more efficiently predict relapse-free survival of esophageal squamous cell carcinoma than existing prognostic factors or than gene expression profiling with cancerous tissue alone.
Asunto(s)
Carcinoma de Células Escamosas/genética , Neoplasias Esofágicas/genética , Esófago/metabolismo , Perfilación de la Expresión Génica , Recurrencia Local de Neoplasia/genética , Anciano , Anciano de 80 o más Años , Neoplasias Esofágicas/mortalidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , PronósticoRESUMEN
BACKGROUND: Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are known to exert a strong adjuvant effect on Th1 immune responses. Although several genes have been reported, no comprehensive study of the gene expression profiles in human cells after stimulation with CpG ODN has been reported. RESULTS: This study was designed to identify a CpG-inducible gene cluster that potentially predicts for the molecular mechanisms of clinical efficacy of CpG ODN, by determining mRNA expression in human PBMC after stimulation with CpG ODN. PBMCs were obtained from the peripheral blood of healthy volunteers and cultured in the presence or absence of CpG ODN 2006 for up to 24 hours. The mRNA expression profile was evaluated using a high-density oligonucleotide probe array, GeneChip. Using hierarchical clustering-analysis, out of a total of 10,000 genes we identified a cluster containing 77 genes as having been up-regulated by CpG ODN. This cluster was further divided into two sub-clusters by means of time-kinetics. (1) Inflammatory cytokines such as IL-6 and GM-CSF were up-regulated predominantly 3 to 6 hours after stimulation with CpG ODN, presumably through activation of a transcription factor, NF-kappaB. (2) Interferon (IFN)-inducible anti-viral proteins, including IFIT1, OAS1 and Mx1, and Th1 chemoattractant IP-10, were up-regulated predominantly 6 to 24 hours after stimulation. Blocking with mAb against IFN-alpha/beta receptor strongly inhibited the induction of these IFN-inducible genes by CpG ODN. CONCLUSION: This study provides new information regarding the possible immunomodulatory effects of CpG ODN in vivo via an IFN-alpha/beta receptor-mediated paracrine pathway.