RESUMO
Although both systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) may lead to joint deformity, SLE arthritis is typically non-erosive and often accompanied by Jaccoud's deformity. Therefore, we examined characteristics of joint and tendon lesions in patients with SLE and RA by ultrasonography. Fifteen treatment-naïve SLE patients and 40 treatment-naïve RA patients with joint symptoms were included in this study. The hand joints and related tendons were ultrasonographically examined using grey-scale (GS) and power Doppler (PD). Joint involvement was comparably observed in patients with SLE and RA (80% versus 95%, p = 0.119). However, tendon involvement was more frequent in SLE than in RA (93% versus 65%, p = 0.045), especially in the wrist joints (73% versus 40%, p = 0.037). When we investigated the intensity of US findings, the joint synovitis score (GS + PD) per affected joint was lower in SLE than RA (2.0 versus 2.6, p = 0.019), while tendon inflammation score was not significantly different (2.1 versus 2.2, p = 0.738). Finally, the examination of concordance between joint and tendon involvement in the same finger revealed that joint lesion appeared in only 49% of fingers having tendon involvement in the SLE group, which was significantly less than 74% in the RA group ( p = 0.010). Thus, as compared with RA, SLE arthropathy is characterized by the predominance of tenosynovitis/periextensor tendon inflammation, which is likely to develop independently from joint synovitis.
Assuntos
Artrite Reumatoide/diagnóstico por imagem , Mãos/diagnóstico por imagem , Lúpus Eritematoso Sistêmico/diagnóstico por imagem , Tendões/diagnóstico por imagem , Adulto , Idoso , Artrite Reumatoide/patologia , Feminino , Humanos , Inflamação/diagnóstico por imagem , Inflamação/etiologia , Lúpus Eritematoso Sistêmico/patologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Sinovite/diagnóstico por imagem , Sinovite/etiologia , Tenossinovite/diagnóstico por imagem , Tenossinovite/etiologia , Ultrassonografia Doppler/métodos , Articulação do Punho/diagnóstico por imagemRESUMO
BACKGROUND: Tumour necrosis factor (TNF) inhibitors enable tight control of disease activity in patients with rheumatoid arthritis (RA). Discontinuation of TNF inhibitors after acquisition of low disease activity (LDA) is important for safety and economic reasons. OBJECTIVE: To determine whether infliximab might be discontinued after achievement of LDA in patients with RA and to evaluate progression of articular destruction during the discontinuation. METHODS: 114 patients with RA who had received infliximab treatment, and whose Disease Activity Score, including a 28-joint count (DAS28) was <3.2 (LDA) for 24 weeks, were studied. RESULTS: The mean disease duration of the 114 patients was 5.9 years, mean DAS28 5.5 and mean modified total Sharp score (mTSS) 63.3. After maintaining LDA for >24 weeks by infliximab treatment, the drug was discontinued and DAS28 in 102 patients was evaluated at year 1. Fifty-six patients (55%) continued to have DAS28<3.2 and 43% reached DAS<2.6 at 1 year after discontinuing infliximab. For 46 patients remission induction by Remicade in RA (RRR) failed: disease in 29 patients flared within 1 year and DAS28 was >3.2 at year 1 in 17 patients. Yearly progression of mTSS (DeltaTSS) remained <0.5 in 67% and 44% of the RRR-achieved and RRR-failed groups, respectively. The estimated DeltamTSS was 0.3 and 1.6 and Health Assessment Questionnaire-Disability Index was 0.174 and 0.614 in the RRR-achieved and RRR-failed groups, respectively, 1 year after the discontinuation. CONCLUSION: After attaining LDA by infliximab, 56 (55%) of the 102 patients with RA were able to discontinue infliximab for >1 year without progression of radiological articular destruction.
Assuntos
Anticorpos Monoclonais/administração & dosagem , Antirreumáticos/administração & dosagem , Artrite Reumatoide/tratamento farmacológico , Adulto , Idoso , Artrite Reumatoide/diagnóstico por imagem , Artrite Reumatoide/fisiopatologia , Avaliação da Deficiência , Progressão da Doença , Esquema de Medicação , Feminino , Seguimentos , Humanos , Infliximab , Masculino , Pessoa de Meia-Idade , Radiografia , Indução de Remissão , Índice de Gravidade de Doença , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adulto JovemRESUMO
We investigated the efficacy and safety of cyclosporine A (CsA; targeted serum trough level: 80-150 ng/ml) in a daily clinical practice for treating patients with systemic lupus erythematosus (SLE), who had been, or were expected to be, refractory to glucocorticoids (GCs) and other immunosuppressants. Fifty-nine patients with SLE receiving CsA were observed for at least 6 months (21.5 months on average). A significant reduction of proteinuria was noted 2 weeks after initiation of treatment in patients with nephritis, resulting in a clinical response in five of eight patients in the GC dose-up group and 11 of 18 patients in the stable GC dose group, respectively. Notably, the mean score for disease activity on the SLE Disease Activity Index decreased significantly from 8.6 +/- 5.3 to 4.4 +/- 2.5 after CsA treatment in patients in the stable GC dose group (n = 40). Moreover, the mean flare rate decreased by approximately 60% with CsA. Side effects of CsA appeared in 32.2% of patients and all of them subsided through dose reduction or discontinuation (n=8) of CsA. Consequently, the cumulative 2-year survival rate of CsA was 75%. The results suggest that CsA should be considered for patients with SLE refractory to GCs.
Assuntos
Ciclosporina/uso terapêutico , Imunossupressores/uso terapêutico , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Nefrite Lúpica/tratamento farmacológico , Adolescente , Adulto , Idoso , Ciclosporina/administração & dosagem , Ciclosporina/efeitos adversos , Relação Dose-Resposta a Droga , Feminino , Seguimentos , Glucocorticoides/uso terapêutico , Humanos , Imunossupressores/administração & dosagem , Imunossupressores/efeitos adversos , Lúpus Eritematoso Sistêmico/mortalidade , Lúpus Eritematoso Sistêmico/fisiopatologia , Nefrite Lúpica/fisiopatologia , Masculino , Pessoa de Meia-Idade , Proteinúria/tratamento farmacológico , Proteinúria/etiologia , Índice de Gravidade de Doença , Taxa de Sobrevida , Resultado do Tratamento , Adulto JovemAssuntos
Anticorpos Monoclonais/efeitos adversos , Síndrome Antifosfolipídica/tratamento farmacológico , Fatores Imunológicos/efeitos adversos , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Trombose Venosa/induzido quimicamente , Doença Aguda , Adulto , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Murinos , Feminino , Humanos , Fatores Imunológicos/uso terapêutico , Rituximab , Adulto JovemRESUMO
OBJECTIVE: To survey and elucidate the clinical characteristics of CMV infection in rheumatic disease patients. METHODS: A detailed questionnaire survey on CMV infection was carried out against rheumatic disease patients hospitalized in member hospitals, and the obtained clinical and/or laboratory data were analysed. RESULTS: Out of 7377 patients, 151 were diagnosed as having CMV infection. The underlying diseases ranged broadly, but SLE, microscopic polyangiitis, and dermatomyositis were the most common. Four were diagnosed histopathologically, and the others via positive CMV antigenaemia. In addition to oral corticosteroid for all but one patient, 81 were treated with pulsed methylprednisolone (MPSL), 64 with cyclophosphamide (CYC) and 36 with other immunosuppressants. Forty-four had a fatal outcome, for which presence of clinical symptoms, other infectious complications, lymphopenia, an older age (>59.3 yrs) and the use of pulsed MPSL were significant risk factors (P < 0.05) by univariate analysis. Multivariate analysis retained the first three (P < 0.05). The CMV antigenaemia count was significantly higher for the symptomatic than asymptomatic [10.1 (0.0-2998.0) vs 4.0 (1.3-1144.4)/10(5) PMNs, respectively, P < 0.05; threshold count: 5.6/10(5) PMNs]. No treatment benefit by anti-viral agent was observed as for survival. CONCLUSION: CMV infection was mostly diagnosed by antigenaemia, and occurred among patients under strong immunosuppressive therapy using pulsed MPSL and/or immunosuppressants. Lymphopenia, presence of symptoms and other infections are significant risk factors for a poor outcome and pulsed MPSL and an older age may predict it. Patients were prone to be symptomatic with anti-genaemia count over 5.6/10(5) PMNs.
Assuntos
Infecções por Citomegalovirus/complicações , Infecções Oportunistas/complicações , Doenças Reumáticas/complicações , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos Virais/sangue , Criança , Ciclofosfamida/administração & dosagem , Citomegalovirus/imunologia , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/epidemiologia , Esquema de Medicação , Feminino , Glucocorticoides/administração & dosagem , Hospitalização , Humanos , Imunossupressores/administração & dosagem , Japão/epidemiologia , Masculino , Metilprednisolona/administração & dosagem , Pessoa de Meia-Idade , Infecções Oportunistas/diagnóstico , Infecções Oportunistas/epidemiologia , Prognóstico , Estudos Retrospectivos , Doenças Reumáticas/tratamento farmacológico , Doenças Reumáticas/epidemiologia , Fatores de RiscoRESUMO
OBJECTIVES: We monitored the mRNA expression profiles of peripheral blood cells during treatment with a TNF-alpha inhibitor, infliximab, in patients with RA. Using a DNA microarray analysis, we demonstrated a unique set of genes, with distinct baseline and post-treatment changes in expression between responders and non-responders to infliximab treatment. METHODS: Using a customized low-density cDNA microarray with 747 genes and a reliable data collection system, we monitored the mRNA expression profiles of whole blood cells from 18 RA patients before and after the infusion of infliximab for up to 22 weeks. The clinical response to treatment with infliximab was determined using the ACR response criteria, the disease activity score of 28 joints (DAS28), and individual clinical parameters. The patients were classified as responders or non-responders based on their ACR50% response at 22 weeks. RESULTS: Approximately 15% of the total genes were found to exhibit a >1.5-fold change, compared with their reference values, at one or more time points during the 22 weeks of infliximab therapy. The expression of inflammatory genes, such as IFN-related genes, was strongly correlated with the serum level of CRP and the DAS28. The increased expression of inflammatory genes in responders was normalized within 2 weeks and then remained at a normal level during the treatment period. In contrast, in the non-responders, the elevated expression at baseline, although it was significantly decreased at 2 weeks, returned to the baseline level after 14 weeks. In addition to inflammatory genes, we identified several groups of genes with distinct differences in expression between the responders and non-responders. CONCLUSIONS: Our results suggest that a customized low-density microarray is useful for monitoring mRNA expression profiles in peripheral blood cells, enabling us to identify a unique set of genes with differentially regulated expressions in responders and non-responders to a TNF inhibitor among patients with RA.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Antirreumáticos/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Regulação da Expressão Gênica/efeitos dos fármacos , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adulto , Idoso , Artrite Reumatoide/sangue , Artrite Reumatoide/genética , Biomarcadores/sangue , Proteína C-Reativa/metabolismo , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , Infliximab , Masculino , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos/métodos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Índice de Gravidade de Doença , Resultado do TratamentoRESUMO
In the field of basic and clinical neurosciences, it is important to develop a method for easy delivery and persistent expression of transgene in central neurons. We firstly generated lentiviral vectors with five kinds of neuron-specific promoters, such as synapsin I (SYN), calcium/calmodulin-dependent protein kinase II, tubulin alpha I, neuron-specific enolase and platelet-derived growth factor beta chain promoters and then novel hybrid promoters by fusing cytomegalovirus enhancer (E) to those neuron-specific promoters. Neuron-specific expression of green fluorescent protein (GFP) with those promoters was examined in vivo by injecting the lentiviral vectors into the rat neostriatum, thalamus and neocortex. Among all the promoters, SYN promoter displayed the highest specificity for neuronal expression in all the regions examined (more than 96%). Although GFP production by the hybrid promoters was about 2-4 times larger than the non-enhanced promoters, the neuronal specificity was significantly decreased in most cases. However, the neuronal specificity of E/SYN hybrid promoter exhibited the least decrease only in the thalamus. Furthermore, the transcriptional activity and neuronal specificity of E/SYN promoter were sustained for up to 8 weeks. Thus, lentivirus with E/SYN promoter is the best vector for strong persistent expression in neurons.
Assuntos
Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , HIV-1/genética , Neurônios/metabolismo , Regiões Promotoras Genéticas , Transdução Genética/métodos , Animais , Encéfalo/metabolismo , Citomegalovirus/genética , Elementos Facilitadores Genéticos , Expressão Gênica , Engenharia Genética , Vetores Genéticos/genética , Proteínas de Fluorescência Verde/genética , Vírus da Hepatite B da Marmota/genética , Injeções , Masculino , Microscopia Confocal , Modelos Animais , Neurônios/virologia , Doença de Parkinson/terapia , Ratos , Ratos Wistar , Sinapsinas/genética , Transfecção/métodosRESUMO
Receptors for platelet-derived growth factor (PDGF) are abundantly expressed on synovial fibroblast-like (SFL) cells from patients with rheumatoid arthritis (RA), and stimulation with PDGF enhances both the anchorage-dependent and -independent growth of RA-SFL cells. To elucidate the molecular mechanisms responsible for the excessive growth of RA-SFL cells and to seek a novel molecular-targeting therapy for RA, we examined the expression of adapter proteins and the effect of the specific inhibition of PDGF receptor activation by imatinib mesylate. Cultured SFL cells were used in the present study after 2-5 passages. The anchorage-dependent and -independent growth patterns of the SFL cells were evaluated using a tetrazolium-based assay and colony formation in 0.3% agar, respectively. Adapter proteins Gab1 and Gab2 were expressed in RA-SFL cells, and both proteins were rapidly (< 1 min) tyrosine-phosphorylated after the stimulation of RA-SFL cells with 10 ng/ml of PDGF and, to a lesser extent, after stimulation with 100 ng/ml of epidermal growth factor (EGF). The inhibition of PDGF receptor tyrosine kinase activation by 1 microM or less of imatinib mesylate specifically suppressed the PDGF-dependent, but not EGF-dependent, tyrosine phosphorylation of various proteins. Moreover, imatinib mesylate abolished both the anchorage-dependent and -independent proliferation of RA-SFL cells induced by PDGF stimulation. These results suggest that Gab adapter proteins are expressed and likely to be involved in the growth signalling of rheumatoid synovial cells and that imatinib mesylate, a key drug in the treatment of chronic myeloid leukaemia, may also be effective for the treatment of RA.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/imunologia , Artrite/imunologia , Fibroblastos/imunologia , Piperazinas/imunologia , Fator de Crescimento Derivado de Plaquetas/imunologia , Inibidores de Proteínas Quinases/imunologia , Pirimidinas/imunologia , Membrana Sinovial/imunologia , Artrite Reumatoide/imunologia , Benzamidas , Divisão Celular/imunologia , Células Cultivadas , Humanos , Mesilato de Imatinib , Osteoartrite/imunologia , Fosforilação , Receptores do Fator de Crescimento Derivado de Plaquetas/imunologia , Transdução de Sinais/imunologia , Membrana Sinovial/patologia , Tirosina/metabolismoRESUMO
An in vitro transformation system of carcinogen-treated Syrian hamster embryo (SHE) cell cultures represents multistep genetic and nongenetic changes that develop during the neoplastic progression of normal cells to tumor cells in vivo. During this neoplastic progression, SHE cells demonstrate an altered response to epidermal growth factor (EGF). In the present report, we examined the role of the adapter protein Gab1 (Grb2-associated binder-1) in the neoplastic progression of SHE cells. We used two asbestos-transformed SHE cell clones in different neoplastic stages: a 10W+8 clone, which is immortal and retains the ability to suppress the tumorigenicity of tumor cells in cell-cell hybrid experiments, and a 10W-1 clone, which has lost this tumor suppressor ability. 10W+8 cells expressed full-length 100-kDa Gab1 and associated 5.2-kb mRNA. Upon repeated cell passaging, 10W-1 cells showed increasing expression of a novel 87-kDa form of Gab1 as well as 4.6-kb mRNA with diminishing expression of the original 100-kDa Gab1. cDNA encoding the 87-kDa Gab1 predicts a form of Gab1 lacking the amino-terminal 103 amino acids (Gab1(Delta1-103)), which corresponds to loss of most of the pleckstrin homology (PH) domain. Gab1(Delta1-103) retains the ability to be phosphorylated in an EGF-dependent manner and to associate with the EGF receptor and SHP-2 upon EGF stimulation. The endogenous expression of Gab1(Delta1-103) in 10W-1 cells appeared closely related to EGF-dependent colony formation in soft agar. Moreover, transfection and expression of Gab1(Delta1-103), but not Gab1, in 10W+8 cells enhanced their EGF-dependent colony formation in soft agar. These results demonstrate that Gab1 is a target of carcinogen-induced transformation of SHE cells and that the expression of a Gab1 variant lacking most of the PH domain plays a specific role in the neoplastic progression of SHE cells.
Assuntos
Proteínas Sanguíneas/química , Neoplasias/metabolismo , Fosfoproteínas/biossíntese , Fosfoproteínas/química , Ágar/metabolismo , Sequência de Aminoácidos , Animais , Amianto , Sequência de Bases , Northern Blotting , Western Blotting , Carcinógenos , Divisão Celular , Linhagem Celular , Células Cultivadas , Clonagem Molecular , Cricetinae , DNA Complementar/metabolismo , Progressão da Doença , Fator de Crescimento Epidérmico/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Sistema de Sinalização das MAP Quinases , Mesocricetus , Dados de Sequência Molecular , Neoplasias/induzido quimicamente , Fenótipo , Fosforilação , Plasmídeos/metabolismo , Testes de Precipitina , Estrutura Terciária de Proteína , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Fatores de Tempo , Transfecção , Transformação GenéticaRESUMO
The adaptor protein Grb2-associated binder-1 (Gab1) is known to bind to the SHP-2 tyrosine phosphatase on epidermal growth factor (EGF) receptor stimulation. To clarify the roles of these two proteins in EGF receptor (EGFR) signaling and determine their possible alteration during neoplastic cell progression, we studied these proteins in a Syrian hamster embryo (SHE) cell line model of neoplastic progression. Specifically, we used asbestos-transformed SHE fibroblasts: the 10W+8 clone, which is immortal but nontumorigenic; and the 10W2T clone, which is tumorigenic. Gab1 was detected, and the EGF-dependent formation of the EGFR-Gab1-SHP-2 complex was observed in 10W+8 cells. After cloning hamster Gab1 cDNA, exogenous expression of Gab1 significantly enhanced EGF-dependent mitogenic activity in 10W+8 cells. On the other hand, Gab1 was not detected in 10W2T cells, and the EGF-dependent association of SHP-2 with EGFR was also absent. Exogenous Gab1 expression in transfected 10W2T cells restored the EGF-dependent association of SHP-2 with EGFR, although it only showed a marginal effect on EGF-dependent mitogenic activity. Thus, Gab1 plays a pivotal role in the EGFR signaling pathway via the formation of the EGFR-Gab1-SHP-2 complex, and alteration in the expression and function of Gab1 is implicated in the neoplastic progression of SHE cells.
Assuntos
Receptores ErbB/metabolismo , Fosfoproteínas/metabolismo , Proteínas Tirosina Fosfatases/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Sequência de Aminoácidos , Animais , Sequência de Bases , Divisão Celular , Clonagem Molecular , Cricetinae , DNA Complementar , Fator de Crescimento Epidérmico/metabolismo , Fator de Crescimento Epidérmico/farmacologia , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Mesocricetus , Camundongos , Mitógenos/metabolismo , Mitógenos/farmacologia , Dados de Sequência Molecular , Fosfoproteínas/genética , Fosfoproteínas/fisiologia , Fosforilação , Proteína Tirosina Fosfatase não Receptora Tipo 11 , Proteína Tirosina Fosfatase não Receptora Tipo 6 , Proteínas Tirosina Fosfatases/fisiologia , Homologia de Sequência de Aminoácidos , Células Tumorais Cultivadas , Tirosina/metabolismoRESUMO
Cyclooxygenase-2 (COX-2) expression is up-regulated in colorectal cancer tissue. Peroxisome proliferator-activated receptors (PPARs) are expressed in human colorectal tissue and activation of PPARs can alter COX-2 expression. In macrophages, activation of PPARs down-regulates COX-2 expression. We examined the effect of PPARalpha and PPARgamma ligands on untreated and TNF-alpha-induced COX-2 expression in the human colorectal epithelial cell line HT-29. The expression of PPARalpha and PPARgamma was confirmed in these cells. TNF-alpha, an inflammatory cytokine, increased COX-2 expression via the NFkappaB pathway. In the absence of TNF-alpha, WY14643 (PPARalpha activator) caused an increase, while BRL49653 (PPARgamma activator) did not alter COX-2 expression. When HT-29 cells were incubated with TNF-alpha and WY14643, a further increase in COX-2 expression was detected. Incubation with TNF-alpha and BRL49653 caused an additional twofold increase in COX-2 expression. Our results suggest that both PPARalpha signaling and TNF-alpha signaling increase COX-2 expression by independent pathways, while PPARgamma stimulates COX-2 expression by up-regulation of the TNF-alpha pathway.
Assuntos
Adenocarcinoma/enzimologia , Neoplasias Colorretais/enzimologia , Isoenzimas/biossíntese , Prostaglandina-Endoperóxido Sintases/biossíntese , Receptores Citoplasmáticos e Nucleares/agonistas , Tiazolidinedionas , Fatores de Transcrição/agonistas , Fator de Necrose Tumoral alfa/farmacologia , Ciclo-Oxigenase 2 , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Células HT29 , Humanos , Isoenzimas/genética , Ligantes , Proteínas de Membrana , Modelos Biológicos , Prostaglandina-Endoperóxido Sintases/genética , Pirimidinas/farmacologia , Receptor Cross-Talk , Rosiglitazona , Transdução de Sinais , Tiazóis/farmacologiaRESUMO
The data presented implicate a GATA binding site in the transcriptional regulation of 15-lipoxygenase-1 (15-LO-1) gene expression in human colorectal carcinoma Caco-2 cells. High expression of GATA-6 mRNA and protein was observed, while GATA-4 mRNA was expressed at a very low level in Caco-2 cells. The expression of GATA-6 was down-regulated, while 15-LO-1 expression was dramatically up-regulated after treatment with sodium butyrate (NaBT). A study using an electrophoretic mobility shift assay indicated that a GATA binding site of the 15-LO-1 promoter region binds to GATA proteins present in both undifferentiated and, to a lesser extent, NaBT-treated (differentiated) Caco-2 cells. Moreover, that DNA binding shift band was disrupted after the addition of GATA-6 antibody in a supershift assay in the absence of NaBT, suggesting that GATA-6 is bound to the GATA binding site of the 15-LO-1 promoter in undifferentiated cells. In contrast, the addition of GATA-6 antibody did not affect the DNA binding ability in NaBT-induced differentiated cells. On the other hand, mutation of the GATA site of the 15-LO-1 promoter decreased the transactivation of the 15-LO-1 promoter as measured by luciferase activity in both FBS and NaBT cultured cells, indicating an unknown GATA binding protein to up-regulate 15-LO-1 expression. These implicate the GATA site at -240 of the proximal region of the 15-LO-1 promoter in the basic transcription of 15-LO-1 gene expression in Caco-2 cells, with GATA-6 acting to repress 15-LO-1 expression.
Assuntos
Araquidonato 15-Lipoxigenase/metabolismo , Proteínas de Ligação a DNA/metabolismo , Fatores de Transcrição/metabolismo , Araquidonato 15-Lipoxigenase/genética , Sítios de Ligação , Western Blotting , Células CACO-2 , Carcinoma , Diferenciação Celular , Neoplasias Colorretais , Proteínas de Ligação a DNA/genética , Fatores de Ligação de DNA Eritroide Específicos , Fator de Transcrição GATA4 , Fator de Transcrição GATA6 , Regulação da Expressão Gênica , Humanos , Regiões Promotoras Genéticas , RNA Mensageiro/metabolismo , Fatores de Transcrição/genéticaRESUMO
In previous studies with Syrian hamster embryo fibroblasts, we found that a specific lipoxygenase metabolite of linoleic acid, 13(S)-hydroperoxyoctadecadienoic acid (HpODE), enhanced epidermal growth factor (EGF) signal transduction in a tumor suppressor gene plus phenotype (supB+); with a diminished response to 13(S)-HpODE in a tumor suppressor gene minus phenotype (supB-). This differential response was attributed to differences in the rate of EGF receptor (EGFR) dephosphorylation. To further define the molecular basis for these observations, in this report we examine the interaction of phosphorylated EGFR with the SH2 domain-containing protein tyrosine phosphatase, SHP-2, a positive modulator of EGF dependent cell growth. SHP-2 associated with phosphorylated EGFR to a greater extent in supB+ cells when compared to supB-. This differential association could not be accounted for by differences between suppressor gene phenotypes in SHP-2 protein level or mutations in the molecular sequence. The addition of 13(S)-HpODE stimulated a concentration-dependent increase in EGF-dependent phosphorylation of SHP-2 and its association with EGFR. A more dramatic response was observed in the supB+ cells. Differences in SHP-2 interaction with EGFR may account, in part, for phenotypic differences in the growth rates and responsiveness to EGF between the supB+ and supB- cells. EGFR-SHP-2 association appears to play an important role in the regulation of EGFR signal transduction.
Assuntos
Receptores ErbB/metabolismo , Ácidos Linoleicos/farmacologia , Peróxidos Lipídicos/farmacologia , Proteínas Tirosina Fosfatases/metabolismo , Animais , Anticorpos , Western Blotting , Linhagem Celular , Genes Supressores de Tumor/genética , Peptídeos e Proteínas de Sinalização Intracelular , Fenótipo , Fosforilação/efeitos dos fármacos , Fosfotirosina/imunologia , Ligação Proteica , Proteína Tirosina Fosfatase não Receptora Tipo 11 , Proteína Tirosina Fosfatase não Receptora Tipo 6 , Proteínas Tirosina Fosfatases Contendo o Domínio SH2 , Domínios de Homologia de srcRESUMO
A role for K+ and Ca2+ channel blockers in cardiac contractile dysfunction and myocardial ionic imbalance was examined in isolated rat hearts with 35-min ischemia and 60-min reperfusion. The K+ channel blockers glibenclamide (1-30 microM) and sematilide (1-30 microM), Ca2+ channel blockers diltiazem (0.1-3 microM) and nicardipine (0.03-1 microM) and fast Na+ channel blocker tetrodotoxin (0.01-0.3 microM) were delivered for the last 3-min pre-ischemia. Ischemia-induced increase in Na+ content was attenuated by diltiazem and tetrodotoxin at all concentrations employed and by nicardipine at 0.3 microM, whereas the ischemia-induced loss of K+ was suppressed partially by glibenclamide and sematilide and almost completely by the two drugs in combination. Left ventricular developed pressure of untreated hearts did not recover upon reperfusion, which was associated with increases in myocardial Na+ and Ca2+ contents and decreases in K+ and Mg2+ contents. Glibenclamide and sematilide neither enhanced the post-ischemic recovery of left ventricular developed pressure nor affected cation changes during reperfusion. Diltiazem enhanced the recovery of left ventricular developed pressure and attenuated imbalance of the myocardial Na+ during ischemia and of all myocardial cations examined during reperfusion. The effects of nicardipine on these parameters were small. Tetrodotoxin enhanced the recovery of left ventricular developed pressure and reversed the imbalance of all myocardial cations examined during reperfusion in a concentration-dependent manner. The results suggest that blockade of transmembrane flux of K+ during ischemia plays a minor role in the improvement of post-ischemic contractile recovery, rather blockade of transmembrane flux of Na+ attenuates the ischemia and reperfusion injury.
Assuntos
Bloqueadores dos Canais de Cálcio/farmacologia , Cátions/metabolismo , Traumatismo por Reperfusão Miocárdica/metabolismo , Miocárdio/metabolismo , Animais , Antiarrítmicos/farmacologia , Creatina Quinase/efeitos dos fármacos , Creatina Quinase/metabolismo , Diltiazem/farmacologia , Relação Dose-Resposta a Droga , Glibureto/farmacologia , Ventrículos do Coração/efeitos dos fármacos , Ventrículos do Coração/fisiopatologia , Hipoglicemiantes/farmacologia , Técnicas In Vitro , Masculino , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Nicardipino/farmacologia , Procainamida/análogos & derivados , Procainamida/farmacologia , Ratos , Ratos Wistar , Tetrodotoxina/farmacologiaRESUMO
OBJECT: To examine the role of human leukocyte antigen (HLA) class II genes in the development of systemic sclerosis (SSc) as well as in the clinical and serologic expression of SSc in patients. METHODS: HLA-DRB1, DRB3, DRB4, DQB1, and DPB1 alleles were determined by genotyping; and serum antinuclear antibodies were identified using indirect immunofluorescence, double immunodiffusion and immunoprecipitation. PATIENTS: One hundred and five Japanese patients with SSc and 104 race-matched healthy controls. RESULTS: Frequencies of DRB1 and DQB1 alleles were not different between SSc patients and healthy controls, while DPB1*0901 was marginally increased in SSc patients. In contrast, SSc-related autoantibodies were closely associated with the clinical features. HLA class II genes were detected as follows: anti-DNA topoisomerase I antibody with diffuse cutaneous involvement, pulmonary fibrosis, and DRB1*1502-DQB1*0601-DPB1*0901; anti-U1RNP antibody with overlapping features of lupus and/or myositis and DRB1*0401/*0802-DQB1*0302; and anticentromere antibody with limited cutaneous involvement and DRB1*0101-DQB1*0501-DPB1*0402. In the analysis of the association of HLA class II and the clinical features in SSc patients significant differences were obtained only for the increased frequencies of arthritis and rheumatoid factor in patients with DRB1*0405 compared to those without. CONCLUSION: HLA class II genes strongly influence the production of SSc-related autoantibodies rather than the development of SSc. In addition, SSc is a composite disease of distinctive subsets defined by serum autoantibodies, which have specific clinical and HLA class II associations.
Assuntos
Autoanticorpos/sangue , Genes MHC da Classe II/fisiologia , Predisposição Genética para Doença , Escleroderma Sistêmico/genética , Escleroderma Sistêmico/imunologia , Adolescente , Adulto , Idoso , Centrômero/imunologia , DNA Topoisomerases Tipo I/imunologia , Feminino , Seguimentos , Frequência do Gene , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-IdadeAssuntos
Antiarrítmicos/efeitos adversos , Síndrome Antifosfolipídica/induzido quimicamente , Lúpus Eritematoso Sistêmico/induzido quimicamente , Procainamida/efeitos adversos , Escleroderma Sistêmico/complicações , Síndrome Antifosfolipídica/patologia , Arritmias Cardíacas/tratamento farmacológico , Doenças do Pé/patologia , Gangrena/patologia , Humanos , Lúpus Eritematoso Sistêmico/patologia , Masculino , Pessoa de Meia-Idade , Pancitopenia/induzido quimicamente , Síndrome , Dedos do Pé/patologiaRESUMO
OBJECTIVE: To examine potential interactions between immunoglobulin (Ig)-allotype gene polymorphisms and susceptibility to systemic sclerosis (SSc) as well as serological expression in SSc patients. METHODS: IgG heavy chain allotypes G1M(f, z), G2M(n+, n-), G3M(b, g) and Ig light chain allotype KM(1, (1, 2), 3) were genotyped in 105 Japanese SSc patients and 47 race matched normal controls using polymerase chain reaction (PCR) based methods. Associations of each Ig allotype with SSc related antinuclear antibodies were examined in combination with or without MHC class II alleles. RESULTS: GM/KM genotypic and allelic frequencies were similar in SSc patients and in normal controls. Frequencies of G1M(f) and G2M(n+) were significantly decreased in anticentromere antibody (ACA) positive SSc patients compared with ACA negative SSc patients (p = 0.04 and 0.02, respectively). Conversely, the presence of DQB1*0501 and KM(1, 2) significantly increased the risk of ACA positivity. CONCLUSION: Ig allotype gene polymorphisms were not associated with susceptibility to SSc. Instead, the results suggested that MHC class II and KM genes are associated with autoimmune responses by interactively promoting the production of ACA.
Assuntos
Anticorpos Antinucleares/biossíntese , Centrômero/imunologia , Genes MHC da Classe II/imunologia , Alótipos de Imunoglobulina/genética , Escleroderma Sistêmico/genética , Humanos , Cadeias Pesadas de Imunoglobulinas/genética , Cadeias Leves de Imunoglobulina/genética , Reação em Cadeia da Polimerase , Polimorfismo Genético , Escleroderma Sistêmico/imunologiaRESUMO
A 47 year-old Japanese female who showed transverse myelopathy (TM) due to spinal epidural hematoma diagnosed by MRI in the course of systemic lupus erythematosus (SLE) was reported. She was admitted to Keio University Hospital due to paraplegia, anesthesia of lower extremity, urinary disturbance. Neurological examination revealed transverse disturbance of Th 10. Lumbar spinal cord MRI showed irregular mass that located at epidural region of 9th-11th thoracic vertebrae. When the laminectomy of 9th-11th thoracic vertebrae was performed, hematoma (4.5 cm x 1.5 cm in size) was confirmed and removed completely. Post operative condition was stable and symptoms had been improving gradually. It has been reported that TM associated with SLE was closely related to myelitis. In this case, epidural hematoma was a major cause of TM and MRI was very useful for her diagnosis and treatment. This is the rare case of SLE associated with spinal epidural hematoma and was thought as a important case to consider the cause of neurological complication of SLE.
Assuntos
Hematoma Epidural Craniano/complicações , Lúpus Eritematoso Sistêmico/complicações , Mielite Transversa/etiologia , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
A case-controlled study was performed, based on early active RA patients treated with MTX; Prednisolone (PSL) was also given in sixteen patients (PSL + MTX group) and each of them was matched for age and sex with a control who have never received PSL (MTX group). No significant differences in radiographic progression were found between the 2 groups. Analysis of radiographic parameters showed that CRP, erythrocyte sedimentation rate, and titers of serum rheumatoid factors after 6 months treatment and their integral amounts during cource were significantly high in the patients with marked radiographic progression. There was no relationship between radiographic progression and treatment with PSL. These results suggested that the indication of PSL therapy for RA is limited for patients with the poor decrease in the level of CRP, erythrocyte sedimentation rate, and rheumatoid factor by MTX treatment.
Assuntos
Anti-Inflamatórios/administração & dosagem , Antirreumáticos/administração & dosagem , Artrite Reumatoide/diagnóstico por imagem , Artrite Reumatoide/tratamento farmacológico , Metotrexato/administração & dosagem , Prednisolona/administração & dosagem , Estudos de Casos e Controles , Quimioterapia Combinada , Feminino , Mãos/diagnóstico por imagem , Humanos , Masculino , Pessoa de Meia-Idade , Radiografia , Estudos RetrospectivosRESUMO
A 68-year-old woman first noticed Raynaud's phenomenon at the age of 35, and she was diagnosed as having systemic sclerosis (SSc) because of cutaneous calcinosis, sclerodactyly, telangiectasia, sausage-like finger, digital pitting scar in 1971. In October, 1994, she was suffered from cold sensation in her right foot, and on November 2 she noticed the painful ulcers on the right toe, which progressed to be gangrenous. Every antiplatelet or thrombolytic agents including PGE1, argatroban, sarpogrelate hydrochloride and urokinase were unsuccessful. Angiograms revealed marked narrowing of bilateral anterior and posterior tibial and peroneal arteries. Prednisolone 30 mg/day was started, which resulted in successful response within 10 days. Large vessel involvement in SSc patients has been reported to be rare, and its treatment remains to be established. We believe our case help understanding the pathophysiology of such rare manifestation in SSc.