RESUMO
Mucosa-associated lymphoid tissue (MALT) lymphomas usually involve extranodal sites, especially the stomach, lung and salivary glands. The Bcl10 gene was recently isolated from the breakpoint region of t(1;14) (p22;q32) in MALT lymphomas, and considered to be an apoptosis-associated gene, and involves a caspase recruitment domain (CARD)-containing protein that activates NF-kappaB. We investigated the role of Bcl10 in MALT lymphoma by analyzing its expression, rearrangement and somatic mutation, by immunostaining, reverse transcriptase-polymerase chain reaction (RT-PCR), Southern blot and PCR in 20 cases of MALT lymphoma. Expression of NF-kappaB was studied by immunostaining. Five cases of reactive lymphadenitis (RLA) were used as the control. Bcl10 rearrangement was detected in 8 of 20 (40%) MALT lymphomas, but in none of RLA. Significant Bcl10 mutation was detected only in 1 case (5%) with MALT, but not in RLA. RT-PCR showed higher density bands of Bcl10 in MALT lymphomas than in RLA. Immunostaining showed a weak Bcl10 expression in the germinal center and very weak expression in the marginal zone B-cells in RLA, which was limited to the cytoplasm. In contrast, Bcl10 was strongly expressed in MALT lymphomas, and was mainly detected in the cytoplasm, as well as in the nuclei. Bcl10 expression did not correlate with Bcl10 mutation and re-arrangements. NF-kappaB was expressed in nuclei of MALT lymphoma cells, but not in RLA. Bcl10 expression in MALT lymphoma correlated closely with NF-kappaB expression. Our results suggest that activation of Bcl10 and NF-kappaB may be important in MALT lymphomagenesis, and that nuclear localization of Bcl10 may be important in the progression of MALT.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal , Linfoma de Zona Marginal Tipo Células B/genética , Proteínas de Neoplasias/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Substituição de Aminoácidos , Proteína 10 de Linfoma CCL de Células B , Southern Blotting , Análise Mutacional de DNA , DNA de Neoplasias/química , DNA de Neoplasias/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Rearranjo Gênico , Humanos , Imuno-Histoquímica , Linfoma de Zona Marginal Tipo Células B/metabolismo , Linfoma de Zona Marginal Tipo Células B/patologia , Masculino , Pessoa de Meia-Idade , Mutação , NF-kappa B/análise , NF-kappa B/genética , Proteínas de Neoplasias/análise , Mutação Puntual , Polimorfismo Conformacional de Fita Simples , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
PURPOSE: To compare the effects of photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) on corneal sensation. SETTING: Ohshima Hospital of Ophthalmology, Fukuoka, Japan. METHODS: Corneal sensation was measured with a Cochet-Bonnet esthesiometer in 35 patients before and 3 days, 1 week, and 1 and 3 months after correction of myopia by PRK (22 patients) or LASIK (13 patients). RESULTS: After PRK, corneal sensitivity was decreased slightly at 3 days, began to recover at 1 week, and returned to preoperative values at 3 months; none of the changes was statistically significant (P >.05). After LASIK, corneal sensation was significantly decreased at 3 days, 1 week, and 1 month; it recovered slightly at 3 months, although it remained significantly less than preoperatively. CONCLUSIONS: Laser in situ keratomileusis was associated with a negative effect on corneal sensation, which was markedly greater than the effect with PRK and was evident for at least 3 months after surgery.
Assuntos
Doenças da Córnea/etiologia , Ceratomileuse Assistida por Excimer Laser In Situ/efeitos adversos , Miopia/cirurgia , Ceratectomia Fotorrefrativa/efeitos adversos , Transtornos de Sensação/etiologia , Adolescente , Adulto , Doenças da Córnea/diagnóstico , Doenças da Córnea/fisiopatologia , Técnicas de Diagnóstico Oftalmológico , Feminino , Humanos , Lasers de Excimer , Masculino , Pessoa de Meia-Idade , Transtornos de Sensação/diagnóstico , Transtornos de Sensação/fisiopatologia , Fatores de TempoRESUMO
Placental leucine aminopeptidase (P-LAP) which is identical with cystine aminopeptidase as oxytocinase was found to be a homologue of rat insulin-regulated membrane aminopeptidase (IRAP) by cDNA cloning. In this study, we confirmed 5'-end cDNA sequence of P-LAP and isolated genomic clones containing the upstream region of human P-LAP gene. The transcription initiation sites determined by primer extension located 478 and 480 bp upstream of the initiation methionine codon, 38 bp downstream of TATA box-like motif. The 5'-flanking region of human P-LAP gene contained DNA-binding motifs for several ubiquitous transcription factors such as SP1 and AP2. Chromosomal localization by fluorescence in situ hybridization showed that the gene was assigned to 5q14.2-q15 of the human chromosome. This study establishes the genetic basis for P-LAP gene research, thereby leading to better understanding of the molecular mechanism underlying the P-LAP gene.
Assuntos
Aminopeptidases/genética , Cistinil Aminopeptidase/genética , Mapeamento Físico do Cromossomo , Regiões Promotoras Genéticas/genética , Animais , Sequência de Bases , Cromossomos Humanos Par 5/genética , Clonagem Molecular , Códon de Iniciação/genética , Biblioteca Genômica , Humanos , Hibridização in Situ Fluorescente , Dados de Sequência Molecular , Placenta/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/genética , Ratos , Elementos de Resposta/genética , Alinhamento de Sequência , TATA Box/genéticaRESUMO
We analyzed the efficacy of all-trans retinoic acid (ATRA) as an early treatment for four acute promyelocytic leukemia (APL) patients in remission who were PML/RARalpha-positive by reverse transcription-polymerase chain reaction or fluorescence in situ hybridization. ATRA 45 mg/m2 was administered orally. All became negative for PML/RARalpha transcripts after 3 to 6 months of ATRA treatment. However, the PML/RARalpha transcripts subsequently reverted to positive in three cases. Although retreatment with ATRA failed to prevent hematological relapse in two patients, one case remains in hematological remission. No serious side effects were encountered during ATRA treatment. These findings suggest that early treatment of ATRA for PML/RARalpha-positive APL patients in remission may have a therapeutic benefit and prolong the duration of hematological remission without chemotherapy.
Assuntos
Antineoplásicos/uso terapêutico , Leucemia Promielocítica Aguda/tratamento farmacológico , Tretinoína/uso terapêutico , Humanos , Hibridização in Situ Fluorescente , Leucemia Promielocítica Aguda/metabolismo , Proteínas de Neoplasias/biossíntese , Proteínas de Fusão Oncogênica/biossíntese , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Although ultraviolet B (UVB) irradiation induces local immune or systemic immune suppression, depending on the dose, the immune suppression by ultraviolet A (UVA) has not been fully investigated. In this study, we investigated the effect of UVA on the immune response in vitro and in vivo. The effect of UVA on the antigen-presenting function of epidermal cells was measured in terms of antigen-specific T cell proliferation. A murine epidermal cell suspension was exposed to UVA in vitro, pulsed with trinitrobenzenesulfonic acid, and cultured with T cells prepared from syngeneic mice previously sensitized with trinitrochlorobenzene. UVA (5-20 J per cm2) suppressed the antigen-presenting function of epidermal cells in a dose-dependent manner, accompanied with suppression of the expression of costimulatory molecules on Langerhans cells. In order to investigate the effect of an antioxidant on the immune suppression, an epidermal cell suspension was irradiated with UVA in the presence or absence of glutathione. The suppressions of antigen-presenting function and ICAM-1 expression were significantly prevented by glutathione in a dose-dependent manner. Further, the effect of UVA on the immune response at the induction phase of contact hypersensitivity was evaluated in terms of lymph node cell proliferation ex vivo. UVA irradiation suppressed the endogenous proliferation of lymph node cells in trinitrochlorobenzene-painted mice, and this suppression was significantly reversed by the application of glutathione to the skin during irradiation. These results suggest that UVA-induced immune suppression may be mediated by reactive oxygen species, at least in part.
Assuntos
Tolerância Imunológica/efeitos da radiação , Espécies Reativas de Oxigênio , Raios Ultravioleta , Animais , Apresentação de Antígeno/efeitos da radiação , Relação Dose-Resposta à Radiação , Feminino , Glutationa/farmacologia , Molécula 1 de Adesão Intercelular/biossíntese , Queratinócitos/efeitos da radiação , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C3HRESUMO
The oxidative effects of cigarette smoke on the human skin were investigated. A remarkable increase in the conversion ratio of squalene (SQ) to squalene monohydroperoxide (SQHPO) due to exposure to cigarette smoke was observed using a CL-HPLC (high performance liquid chromatography with a chemiluminescence detector) system. The results showed that cigarette smoke caused lipid peroxidation. We also found that the addition of chain-breaking-type antioxidants, such as oolong tea extract, inhibited the peroxidation. When cultured human skin fibroblasts were exposed to cigarette smoke, this increased the intensity of ultraweak chemiluminescence (CL), leading us to assume that cigarette smoke caused oxidation in cultured human skin fibroblasts. When the cultured human skin fibroblasts were treated with antioxidants such as glutathione, thiotaurine, hypotaurine and ascorbic acid there was little increase in CL, meaning that oxidation had been prevented in the human skin fibroblasts. We also exposed the human forearm to cigarette smoke and obtained sebum using cotton immersed in acetone in order to measure hydroperoxide levels by means of a CL-HPLC system. The exposure of skin to the smoke caused a dose-dependent increase in hydroperoxides derived from cigarette smoke. Further exposure of the forearm to cigarette smoke increased the intensity of CL, but pretreating the skin with antioxidants such as glutathione, thiotaurine and hypotaurine inhibited this increase. From these results, we concluded that cigarette smoke had an oxidative effect on SQ, cultured human skin fibroblasts and the surface of the human skin. The application of antioxidants prevented the cigarette smoke-induced oxidation. We consider that these oxidative effects on the skin could be a cause of skin disorders and skin aging.
RESUMO
BACKGROUND: The focus of these studies was to determine whether the Y79 human retinoblastoma cell line could function as a good in vitro model system for studying human cytomegalovirus (HCMV) infection. METHODS: Y79 cells were exposed to an HCMV mutant carrying a LacZ gene, and the resulting beta-galactosidase expression in infected cells was assessed by flow cytometry. The extent to which the three classes of viral gene products immediate early, early, and late proteins - were expressed was analyzed by immunohistochemical staining and Western blotting. Infected Y79 cells were also co-cultivated on human foreskin fibroblast (SF cell) cultures to recover virus. RESULTS: Infection of Y79 cells with the virus resulted in beta-galactosidase expression as detected by flow-cytometric analysis. Immunohistochemical staining revealed that a portion of Y79 cells expressed antigens reactive to monoclonal antibodies against immediate early, early, and late HCMV proteins. The 43-kDa early gene product was also detected by Western blotting. Infected Y79 cells co-cultivated on SF cell cultures yielded infectious foci, which turned blue following X-gal staining, demonstrating productive HCMV infection in the Y79 cells. CONCLUSION: These results demonstrate that while HCMV can productively infect Y79 cultures, it does so in a highly inefficient manner, leading these authors to conclude that this cell line does not provide a particularly good model system to study HCMV infection.
Assuntos
Citomegalovirus/fisiologia , Neoplasias da Retina/virologia , Retinoblastoma/virologia , Western Blotting , Citometria de Fluxo , Seguimentos , Galactosídeos/biossíntese , Galactosídeos/genética , Genes Precoces/genética , Genes Virais/genética , Humanos , Técnicas Imunoenzimáticas , Óperon Lac/fisiologia , RNA Viral/análise , Neoplasias da Retina/metabolismo , Neoplasias da Retina/patologia , Retinoblastoma/metabolismo , Retinoblastoma/patologia , Células Tumorais Cultivadas/virologia , Proteínas Virais/análiseRESUMO
The biological activity of the novel vitamin C derivative, 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G), was evaluated in vitro and in vivo. The percutaneous absorption of AA-2G was determined in five Japanese males. The excretion of ascorbic acid (AA) in the subjects administered AA-2G was sustained for a longer period than in the subjects administered ascorbic acid 2-phosphate (AA-2P), which is a conventional vitamin C derivative. An analysis of the distribution of AA in the skin showed that small black specks assumed to be AA were observed in the epidermis even 3 d after applying AA-2G. The melanin synthesis in B16 melanoma cells was inhibited more by AA-2G than by AA-2P, and AA-2G also prevented more UV-induced damage of human skin keratinocytes and fibroblasts than AA-2P did. From these in vivo and in vitro results, it is supposed that the conversion of AA-2G to AA is sustained for a long time compared with that of AA-2P, and that AA-2G is an effective and available compound having vitamin C activity in human subjects.
Assuntos
Ácido Ascórbico/análogos & derivados , Cosméticos , Absorção , Ácido Ascórbico/análise , Ácido Ascórbico/metabolismo , Ácido Ascórbico/farmacocinética , Ácido Ascórbico/farmacologia , Ácido Ascórbico/uso terapêutico , Células Cultivadas , Epiderme/metabolismo , Humanos , Radical Hidroxila/metabolismo , Queratinócitos/efeitos dos fármacos , Queratinócitos/efeitos da radiação , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Melaninas/biossíntese , Melanoma/metabolismo , Pele/química , Pele/metabolismo , Queimadura Solar/prevenção & controle , Distribuição Tecidual , Células Tumorais Cultivadas , Raios UltravioletaRESUMO
The purpose of this study is to evaluate hemodynamic characteristics of various hepatic tumors using color Doppler echography administered galactose-based intravenous contrast medium "SH/TA 508 (Levovist)". Subject were 9 cases of hepatocellular carcinomas, 5 cases of metastatic liver tumors and a case of hemangioma. We evaluated the characteristics of blood flows inside various hepatic tumors, and also evaluated the first pass through the hepatic tumors during administration of Levovist. The strongly pulsatile branched blood flows inside tumor and the slow-increased and plateau patterns were observed in the all cases of hepatocellular carcinomas, the weakly pulsatile blood flows surrounding tumor and the rapid-increased and slow-decreased patterns were observed in the cases of metastatic liver tumors, and the continuous blood streams in the margin of tumor and the slow-increased and slow-decreased pattern were observed in the case of hemangioma. These findings were characteristic in various hepatic tumors, and color Doppler echography enhanced by Levovist was very useful to distinguish hepatic tumors.
Assuntos
Meios de Contraste/administração & dosagem , Neoplasias Hepáticas/irrigação sanguínea , Neoplasias Hepáticas/diagnóstico por imagem , Polissacarídeos/administração & dosagem , Ultrassonografia Doppler em Cores/métodos , Carcinoma Hepatocelular/irrigação sanguínea , Carcinoma Hepatocelular/diagnóstico por imagem , HumanosRESUMO
The Wnt genes compose a large gene family encoding a group of secreted signaling molecules that have been implicated in oncogenesis and a number of developmental processes. We have isolated a full-length human cDNA clone that we consider to be a novel member of the Wnt gene family. The gene (WNT7A) encodes a deduced 349-amino-acid peptide with 98% identity in amino acid sequence to murine Wnt7a. Expression of this gene is restricted to certain tissues: placenta, kidney, testis, uterus, fetal lung, and fetal and adult brain. Furthermore, we have isolated a genomic clone of WNT7A and mapped it to chromosome 3p25 by fluorescent in situ hybridization.
Assuntos
Mapeamento Cromossômico , Cromossomos Humanos Par 3 , Proteínas/genética , Proteínas Proto-Oncogênicas , Sequência de Aminoácidos , Sequência de Bases , Northern Blotting , Clonagem Molecular , DNA Complementar/genética , Feminino , Expressão Gênica/genética , Humanos , Hibridização in Situ Fluorescente , Dados de Sequência Molecular , Proteínas WntRESUMO
Recently three strains of herpes simplex virus type 1 (HSV-1), which did not react with Micro Trak Herpes (Syva Co.), were isolated by us from a patient with recurrent herpetic keratitis. In this study we characterized these strains of HSV-1 and found them to be HSV-1 gC- mutants which are very rare isolates from humans. The properties of the HSV-1 strains regarding plaque morphology on Vero cells and chick embryo fibroblasts and viral DNA analysis were the same as those of the usual HSV-1 strains. An immunofluorescence study using anti-gC-1 monoclonal antibody and SDS-PAGE analysis of radiolabeled viral glycoproteins showed that these strains are deficient in gC-1. They were virulent for mice and sensitive to acyclovir and bromovinyldeoxyuridine. Furthermore the infectivity of the strains was inactivated by complement though the phenomenon was not observed in the usual HSV-1 strains. This finding suggests that protection from damages by complement is an important function of gC. In keratitis the effects of complement are thought to be minimal because of the scanty blood supply and this may be the reason why these strains were isolated from the cornea.
Assuntos
Ceratite Dendrítica/microbiologia , Mutação , Simplexvirus/genética , Adulto , Animais , Proteínas do Sistema Complemento , Enzimas de Restrição do DNA , DNA Viral/química , Eletroforese em Gel de Poliacrilamida , Imunofluorescência , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Simplexvirus/isolamento & purificação , Simplexvirus/metabolismo , Simplexvirus/patogenicidade , Células Vero , Proteínas do Envelope Viral/biossíntese , Ensaio de Placa ViralRESUMO
The effect of monoclonal antibody (MCA) to glycoprotein gB of herpes simplex virus (HSV) was studied in athymic nude mice inoculated with HSV intracutaneously in the midflank. HS1, the MCA used in the study, had a high neutralizing titer (1:2048) and had antibody-dependent cell-mediated cytotoxicity. HS1 was injected intraperitoneally at various intervals after HSV infection. HS1 injected 3 h after infection inhibited the development of skin lesions and most mice survived. Administration of HS1 at the time the local skin erosions appeared at the inoculated site (4-7 days after infection) was also effective, and in four of eight mice skin lesions completely healed. Furthermore, in three of four mice that survived, latent infections in the ganglia were also prevented as evidenced by the failure to detect HSV by co-cultivation with Vero cells. Administration of HS1 after the development of zosteriform skin lesions (5-9 days after infection) reduced virus in the ganglia and prolonged the survival time, though the disease was not completely arrested and all the mice died eventually.