RESUMO
Allogeneic hematopoietic SCT (allo-SCT) is a promising therapy that may provide long-term durable remission for adult T-cell leukemia-lymphoma (ATL) patients; however, the incidence of relapse associated with ATL remains high. To determine the clinical features of these patients at relapse, we retrospectively analyzed tumor lesions in 30 or 49 patients who relapsed following allo-SCT or chemotherapy (CHT), respectively, at three institutions in Nagasaki prefecture between 1997 and 2011. A multivariate analysis revealed that the development of abnormal lymphocytes in the peripheral blood of patients at relapse was less frequent after allo-SCT than after CHT (P<0.001). Furthermore, relapse with a new lesion only in the absence of the primary lesion was more frequent in allo-SCT (P=0.014). Lesions were more frequently observed in the central nervous systems of patients who relapsed with new lesions only (P=0.005). Thus, the clinical manifestation of relapsed ATL was slightly complex, especially in post-transplant patients. Our results emphasized the need to develop adoptive modalities for early and accurate diagnoses of relapsed ATL.
Assuntos
Transplante de Células-Tronco Hematopoéticas , Leucemia-Linfoma de Células T do Adulto , Adulto , Idoso , Idoso de 80 Anos ou mais , Aloenxertos , Feminino , Humanos , Japão , Leucemia-Linfoma de Células T do Adulto/diagnóstico , Leucemia-Linfoma de Células T do Adulto/mortalidade , Leucemia-Linfoma de Células T do Adulto/patologia , Leucemia-Linfoma de Células T do Adulto/terapia , Linfócitos/patologia , Masculino , Pessoa de Meia-Idade , RecidivaRESUMO
Myeloperoxidase (MPO) has been associated with both a myeloid lineage commitment and favorable prognosis in patients with acute myeloid leukemia (AML). DNA methyltransferase inhibitors (decitabine and zeburaline) induced MPO gene promoter demethylation and MPO gene transcription in AML cells with low MPO activity. Therefore, MPO gene transcription was directly and indirectly regulated by DNA methylation. A DNA methylation microarray subsequently revealed a distinct methylation pattern in 33 genes, including DNA methyltransferase 3 beta (DNMT3B), in CD34-positive cells obtained from AML patients with a high percentage of MPO-positive blasts. Based on the inverse relationship between the methylation status of DNMT3B and MPO, we found an inverse relationship between DNMT3B and MPO transcription levels in CD34-positive AML cells (P=0.0283). In addition, a distinct methylation pattern was observed in five genes related to myeloid differentiation or therapeutic sensitivity in CD34-positive cells from AML patients with a high percentage of MPO-positive blasts. Taken together, the results of the present study indicate that MPO may serve as an informative marker for identifying a distinct and crucial DNA methylation profile in CD34-positive AML cells.
Assuntos
DNA (Citosina-5-)-Metiltransferases/genética , Metilação de DNA , Regulação Leucêmica da Expressão Gênica , Leucemia Mieloide Aguda/genética , Peroxidase/genética , Antígenos CD34/metabolismo , Medula Óssea/patologia , Células da Medula Óssea/metabolismo , Células da Medula Óssea/patologia , Proteínas Estimuladoras de Ligação a CCAAT/genética , Linhagem Celular Tumoral , Análise por Conglomerados , DNA (Citosina-5-)-Metiltransferases/metabolismo , Epigênese Genética , Perfilação da Expressão Gênica , Humanos , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patologia , Mutação , Proteínas Nucleares/genética , Nucleofosmina , Peroxidase/metabolismo , Tirosina Quinase 3 Semelhante a fms/genética , DNA Metiltransferase 3BRESUMO
Adult T-cell leukemia/lymphoma (ATLL) is a neoplasia characterized by the massive invasion of various organs by tumor cells. Previously, we found that expression of the gene for c-Met, a receptor tyrosine kinase for hepatocyte growth factor (HGF), was specific to the acute type among 41 patients with ATLL by microarray. First in the present study, we analyzed the survival of the patients in relation to expression of c-Met and HGF in ATLL cells. Expression of the former but not the latter was associated with poor prognosis. Then, we analyzed the growth of ATLL cells caused by HGF and c-Met. c-Met was expressed in 0/7 chronic ATLLs, 12/14 acute ATLLs, 1/1 IL-2-independent ATLL cell line and 1/7 IL-2-dependent ATLL cell lines as assessed by flow cytometry. HGF induced the proliferation of primary cells from most acute cases examined as well as the c-Met-positive KK1 cell line in contrast to c-Met-negative cells. HGF induced autophosphorylation of c-Met in c-Met-positive cells from an acute case and KK1 cells. The plasma level of HGF was elevated in acute as compared to chronic cases. The levels of HGF and/or IL-6 which induces the production of HGF by stromal cells, were elevated in the supernatant of short-term cultured cells from certain patients with acute or chronic disease. Finally, infiltrated ATLL cells and adjacent stromal cells in liver were shown to be positive for c-Met/HGF and HGF, respectively, in acute cases. Autocrine and/or paracrine growth caused by HGF and c-Met was suggested in aggressive ATLL cells secreting HGF and/or IL-6, respectively.
Assuntos
Regulação Leucêmica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Fator de Crescimento de Hepatócito/metabolismo , Leucemia-Linfoma de Células T do Adulto/imunologia , Proteínas Proto-Oncogênicas c-met/metabolismo , Apoptose , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Proliferação de Células , Citocinas/metabolismo , Humanos , Interleucina-6/metabolismo , Leucemia-Linfoma de Células T do Adulto/metabolismo , Modelos Biológicos , Fosforilação , Fatores de TempoRESUMO
Myeloperoxidase (MPO), a pivotal lineage marker for acute myeloid leukemia (AML), has been also shown to have a prognostic value: a high percentage of MPO-positive blasts correlates to favorable prognosis. To understand the relationship between the expression of MPO in leukemia cells and the response to chemotherapeutic agents, we established MPO-expressing K562 leukemia cell lines and then treated them with cytosine arabinocide (AraC). Cells expressing wild-type MPO, but not mutant MPO that could not mature, died earlier of apoptosis than control K562 cells. Reactive oxygen species (ROS) were generated more in leukemia cells expressing MPO, and the generation was abrogated by MPO inhibitors or antioxidants. Tyrosine nitration of cellular protein also increased more in MPO-expressing K562 cells than control cells after treatment with AraC. In clinical samples, CD34-positive AML cells from high-MPO cases showed a tendency to be sensitive to AraC in the colony-formation assay, and the generation of ROS and the nitration of protein were observed only when the percentage of MPO-expressing cells was high. These data suggest that MPO enhances the chemosensitivity of AML through the generation of ROS and the nitration of proteins.
Assuntos
Antineoplásicos/farmacologia , Leucemia/patologia , Peroxidase/fisiologia , Processamento de Proteína Pós-Traducional , Espécies Reativas de Oxigênio/metabolismo , Humanos , Células K562 , Leucemia/metabolismo , Nitrosação , Peroxidase/análise , Células Tumorais CultivadasRESUMO
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) can provide long-term remission for patients with adult T-cell leukemia/lymphoma (ATLL) caused by human retrovirus, human T-lymphocyte virus (HTLV-1). To understand how HTLV-1-positive cells including ATLL cells were suppressed by allo-HSCT, we examined HTLV-1 provirus load and residual ATLL cells in peripheral blood of transplant recipients using PCR-based tests. We found that the copy number of HTLV-1 genome, called provirus, became very small in number after allo-HSCT; however, in most cases, provirus did not disappear even among long-term survivors. Tumor-specific PCR tests demonstrated that most of HTLV-1-positive cells that remained long after transplantation were not primary ATLL cells but donor-derived HTLV-1-positive cells. We also found a case having very low amount of residual disease in peripheral blood even long after transplantation. There was only one recipient in whom we failed to show the presence of HTLV-1 genome and antibody against HTLV-1 even with an extensive search, which strongly suggested the elimination of HTLV-1 after allo-HSCT. These results demonstrated that after allo-HSCT the small amount of residual HTLV-1-positive cells were heterogeneous in origin and that long-term disease control for ATLL could be obtained without the complete elimination of HTLV-1.
Assuntos
Transplante de Células-Tronco Hematopoéticas , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Leucemia-Linfoma de Células T do Adulto/terapia , Adulto , Humanos , Leucemia-Linfoma de Células T do Adulto/patologia , Reação em Cadeia da Polimerase , Indução de Remissão , Doadores de Tecidos , Transplante Homólogo , Carga ViralRESUMO
Tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) induces apoptosis in many transformed cells, but not in normal cells, and hence TRAIL has recently emerged as a novel anti-cancer agent. Adult T-cell leukaemia lymphoma (ATLL) is a neoplasm of T-lymphocyte origin aetiologically associated with human T-lymphotropic virus type 1 (HTLV-I), and is resistant to standard anti-cancer therapy. We thus characterized the sensitivity of ATLL cells to TRAIL in this study. Although most primary ATLL cells and cell lines expressed TRAIL death receptors on their surface, they showed only restricted sensitivity to TRAIL. Among the 10 ATLL cell lines examined, one was sensitive, but two had insufficient death-receptor expression, two had an unknown resistant mechanism with abrogation of the death signal upstream of caspase-8, and the remaining five showed attenuation of the signal in both extrinsic and intrinsic pathways by X-linked inhibitor of apoptosis and Bcl-2/Bcl-xL respectively. Furthermore, the level of HTLV-I tax expression was significantly correlated to TRAIL resistance. Interestingly, ATLL cells themselves expressed TRAIL on the cell surface. Constitutive production of TRAIL may offer resistance, thus allowing the development of TRAIL-resistant ATLL cells. Consequently, the resistant mechanism in ATLL cells against TRAIL was assigned to multiple factors and was not explained by a definitive single agent.
Assuntos
Antineoplásicos/uso terapêutico , Leucemia-Linfoma de Células T do Adulto/tratamento farmacológico , Glicoproteínas de Membrana/uso terapêutico , Fator de Necrose Tumoral alfa/uso terapêutico , Apoptose , Proteínas Reguladoras de Apoptose , Resistencia a Medicamentos Antineoplásicos/genética , Expressão Gênica , Genes bcl-1 , Genes pX , Humanos , Leucemia-Linfoma de Células T do Adulto/genética , Leucemia-Linfoma de Células T do Adulto/patologia , Receptores do Ligante Indutor de Apoptose Relacionado a TNF , Receptores do Fator de Necrose Tumoral/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ligante Indutor de Apoptose Relacionado a TNF , Células Tumorais CultivadasRESUMO
Interferon regulatory factor (IRF) 4 is the lymphoid-specific transcription factor that is required for the proliferation of mitogen-activated T cells. IRF4 has been suggested to be involved in tumorigenesis because the overexpression of IRF4 caused the transformation of Rat-1 fibroblasts in vitro. Here, we show that IRF4 is constitutively expressed in adult T-cell leukemia (ATL)-derived cell lines, which were infected with human T-cell leukemia virus type-I, but hardly expressed the trans-activator protein, Tax. Similarly, constitutive expression of IRF4 was demonstrated in freshly isolated peripheral blood mononuclear cells (PBMC) from patients with either acute or chronic ATL. However, the high-level expression of IRF4 was specifically associated with acute ATL. With mitogen-activated PBMC from healthy donors, cell cycle analyses revealed that the induction of IRF4 occurred prior to cell cycle progression and the cells that had entered the cell cycle were predominantly IRF4-positive cells. In addition, ectopic expression of IRF4 in Rat-1 fibroblasts increased the S and G2 / M phase population significantly. Taken together, our results indicate that IRF4 is involved in the pathogenesis of ATL through its positive effect on the cell cycle, and that IRF4 can be used as a molecular marker of clinical subtype in ATL.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Leucemia-Linfoma de Células T do Adulto/metabolismo , Leucemia-Linfoma de Células T do Adulto/patologia , Fatores de Transcrição/metabolismo , Adulto , Animais , Western Blotting , Ciclo Celular/efeitos dos fármacos , Proteínas de Ligação a DNA/genética , Fibroblastos , Citometria de Fluxo , Regulação da Expressão Gênica/efeitos dos fármacos , Produtos do Gene tax/metabolismo , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Humanos , Fatores Reguladores de Interferon , Células Jurkat , Leucemia-Linfoma de Células T do Adulto/classificação , Leucemia-Linfoma de Células T do Adulto/virologia , Mitógenos/farmacologia , RNA Mensageiro/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição/genética , Células Tumorais CultivadasRESUMO
OBJECTIVE: The gene responsible for hereditary hemochromatosis close to the human leukocyte antigen A locus was previously identified and designated as HFE. This study was performed to evaluate the clinical significance of two mutations, C282Y and H63D of HFE, in Japanese patients with hepatic iron overload. PATIENTS AND METHODS: We examined C282Y and H63D in 11 patients with primary hemochromatosis, 94 patients with chronic hepatitis C, 54 patients with miscellaneous liver diseases, and 151 healthy volunteers. The HFE gene region of DNA samples extracted from peripheral leukocytes was amplified by polymerase chain reaction. Restriction enzyme analysis was performed using SnaBI for C282Y and BclI for H63D. Direct sequence analysis was then performed when products suggested the presence of a mutation. RESULTS: All the subjects studied were free from C282Y. None of the patients with hemochromatosis had H63D. One patient with chronic hepatitis C was homozygous, and 4 patients were heterozygous for H63D. Two patients with alcoholic liver disease were heterozygous for H63D. The prevalence of chromosomes with H63D was 6/188 (3.2%) in patients with chronic hepatitis C, 2/108 (1.9%) in patients with miscellaneous liver diseases, and 8/302 (2.6%) in healthy volunteers. These differences were not significant. CONCLUSION: Our results suggested that neither C282Y nor H63D in HFE affect Japanese patients with hemochromatosis or chronic hepatitis C.
Assuntos
Povo Asiático/genética , Ácido Aspártico/genética , Cisteína/genética , Antígenos HLA/genética , Hemocromatose/genética , Histidina/genética , Antígenos de Histocompatibilidade Classe I/genética , Hepatopatias/genética , Proteínas de Membrana , Mutação Puntual , Tirosina/genética , Adulto , Feminino , Hemocromatose/epidemiologia , Proteína da Hemocromatose , Hepatite C Crônica/genética , Humanos , Sobrecarga de Ferro/genética , Japão/epidemiologia , Hepatopatias/epidemiologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da PolimeraseRESUMO
In order to investigate the mechanism of anti-rheumatic action of mizoribine (MZR), antiproliferative effect of MZR on synovial fibroblasts obtained from rheumatoid arthritis (RA) patients were examined. To examine the effect of MZR on DNA synthesis, total radioactivity of 3H-thymidine (3H-TdR) incorporated into the synovial fibroblasts was measured. Also quantification of DNA fragmentation of synovial fibroblasts in the cultured supernatant and cell associated Bcl-2 protein, which is suspected of interfering with apoptosis, were performed with enzyme-linked immunosorbent assay. MZR suppressed 3H-TdR incorporation into synovial fibroblasts in a dose dependent fashion. Significant inhibition (P < 0.01) was attained at the concentration of more than 1 microgram/ml of MZR. However, induction of DNA fragmentation which is characteristic of apoptosis, were observed at only 10 micrograms/ml of MZR over 72 h-incubation significantly. In terms of the Bcl-2 expression of synovial fibroblasts, up to 10 micrograms/ml of MZR has no effect on the expression of this protooncogene bcl-2 expression. These results suggest that MZR might suppress the growth of rheumatoid pannus by inhibition of synovial fibroblast proliferation partially through the induction of apoptosis of synovial fibroblast without modulating Bcl-2 expression.
Assuntos
Apoptose/efeitos dos fármacos , Artrite Reumatoide/patologia , Divisão Celular/efeitos dos fármacos , Fibroblastos/patologia , Ribonucleosídeos/farmacologia , Membrana Sinovial/patologia , Células Cultivadas , DNA/biossíntese , Fragmentação do DNA , Relação Dose-Resposta a Droga , Humanos , Imunossupressores/administração & dosagem , Imunossupressores/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Ribonucleosídeos/administração & dosagemRESUMO
The effects of KRN2391, an ATP-sensitive K+ channel opener (KCO) which also acts as a nitrate, on ionic membrane currents in rabbit femoral arterial myocytes were examined. Under whole-cell clamp conditions where cells were superfused with physiological salts solution containing 5.9 mM K+, KRN2391 elicited an outward current at a holding potential of -30 mV. KRN2391-induced current had a reversal potential of -78 mV and was abolished by glibenclamide (glib). KRN2391 was approximately 25 times more potent than nicorandil to activate an ATP-sensitive K+ current (I:(KATP)). On the other hand, 10 microM KRN2391 did not affect either voltage-dependent Ca(2+) or delayed rectifier K+ channel currents. In the inside-out patch configuration, KRN2391 activated 47 pS K+ channels in the presence of nucleotide diphosphates (NDPs) under the symmetrical 140 mM K+ conditions. Glib and intracellular ATP reversibly inhibited the activity of the 47 pS K+ channels. The 47 pS K+ channels activated by KRN2391 are similar in their conductance and other properties to NDP-sensitive K+ channels (K(NDP) channels) described in other smooth muscles and the cloned channels. KRN2391 is a potent activator of the 47 pS K+ channels and the activation can contribute to the KRN2391-induced vasodilation in arterial muscles.
Assuntos
Trifosfato de Adenosina/fisiologia , Artéria Femoral/efeitos dos fármacos , Canais de Potássio/efeitos dos fármacos , Piridinas/farmacologia , Vasodilatadores/farmacologia , Difosfato de Adenosina/farmacologia , Animais , Artéria Femoral/fisiologia , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Nicorandil/farmacologia , Canais de Potássio/fisiologia , Coelhos , Difosfato de Uridina/farmacologiaRESUMO
Two young adults underwent resection of large hypervascular vestibular schwannomas (acoustic neuromas) via two-stage surgery. The first patient, a 27-year-old woman with hydrocephalus, had a large hypervascular vestibular tumor in the left cerebellopontine angle (CPA) supplied by the left anterior inferior cerebellar artery (AICA) and posterior inferior cerebellar artery (PICA). The second patient, a 34-year-old woman, had a large AICA-supplied hypervascular vestibular tumor in the left CPA that displaced the brain stem significantly. At the initial stage, only the lateral aspect of the tumor was debulked due to excessive bleeding from the tumor bed. Angiography 1 or 2 months after the initial operation showed that the tumor was hypovascular. At the second stage, the remnant medial aspect of the tumor was relatively avascular and nonadherent to the brain stem. Without blood transfusion during the second stage, the tumor was removed totally in the first patient and subtotally in the second patient. Pathological examination revealed that dilatated blood vessels were prominently increased at the first surgery; however, at the second surgery, the number of blood vessels had decreased, showing necrosis and degeneration. Although there are no absolute indications for the staged resection of vestibular schwannomas, this procedure may represent one of the safest options for these difficult lesions in young adults.
RESUMO
The purpose of this study was to determine whether prostate cancer mortality conforms to gompertzian analysis and, if so, what conclusions can be made regarding increasing prostate cancer mortality in Japan by the application of longitudinal gompertzian analysis. Data regarding 77,492 prostate cancer deaths reported during the period 1955 to 1996 were obtained from death certificate records in Japan. Age-adjusted prostate cancer mortality rates increased 6.4-fold during that period. The changing patterns in the mortality rate was explained by a constantly increasing number of elderly people. Age-specific mortality rate distributions between the ages of 50 and 84 years were highly gompertzian for each year during that period. The environmental factor for prostate cancer mortality increased 80% for that period. Gompertzian analysis suggests that rising mortality from prostate cancer may be related to rapidly changing lifestyles among Japanese. Intake of animal proteins was the most strongly correlated parameter considered with the mortality from prostate cancer.
Assuntos
Neoplasias da Próstata/mortalidade , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Consumo de Bebidas Alcoólicas , Dieta/estatística & dados numéricos , Humanos , Japão/epidemiologia , Estilo de Vida , Modelos Lineares , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Mortalidade/tendências , FumarRESUMO
Ruthenium Red (RuR) is widely used as an inhibitor of ryanodine receptor Ca(2+) release channels, but has additional effects such as the induction of Ca(2+) sensitization of contraction of permeabilized smooth muscles. To address the mechanism underlying this process, we examined the effects of RuR on contractility in permeabilized guinea-pig ileum and on the activity of myosin-light-chain phosphatase (MP). RuR increased the force at submaximal [Ca(2+)] (pCa 6.3) approx. 4-fold. This effect was not observed after thiophosphorylation of MP. RuR also seemed capable of preventing the thiophosphorylation of MP, suggesting a direct interaction of RuR with MP. Consistent with this possibility, smooth-muscle MP was inhibited by RuR in a concentration-dependent manner (IC(50) 23 microM). Exogenous calmodulin significantly increased RuR-induced contraction at pCa 6.3 but had little effect on contraction induced by microcystin at this [Ca(2+)]. Ca(2+)-independent contraction was induced by RuR (EC(50) 843 microM) and by microcystin (EC(50) 59 nM) but the maximal force induced by RuR was smaller than that induced by microcystin. The addition of 300 microM RuR enhanced the contraction induced by 30 nM microcystin but markedly decreased that induced by 1 microM microcystin. Such a dual action of RuR on microcystin-induced effects was not observed in experiments using purified MP. We conclude that the RuR-induced Ca(2+) sensitization of smooth-muscle contraction is due to the direct inhibition of MP by RuR.
Assuntos
Trifosfato de Adenosina/análogos & derivados , Inibidores Enzimáticos/farmacologia , Músculo Liso/enzimologia , Fosfoproteínas Fosfatases/antagonistas & inibidores , Rutênio Vermelho/farmacologia , Trifosfato de Adenosina/farmacologia , Animais , Cálcio/metabolismo , Calmodulina/metabolismo , Cobaias , Técnicas In Vitro , Masculino , Microcistinas , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Músculo Liso/fisiologia , Fosfatase de Miosina-de-Cadeia-Leve , Peptídeos Cíclicos/farmacologia , FosforilaçãoRESUMO
The characteristics of the cromakalim-induced membrane current were examined in single tracheal myocytes of the guinea-pig under voltage-clamp conditions. When K(+) concentrations in the pipette and bathing solutions were approximately 140 mM, cromakalim activated a membrane current (I(crom)) which was inward at -60 mV and reversed at -2 mV. I(crom) was blocked by 10 microM glibenclamide and potentiated when the ATP concentration in the pipette solution was decreased. The K(d) and Hill coefficient of glibenclamide for I(crom) block were 200 nM and 1.05, respectively. Application of the tyrosine kinase inhibitors, genistein and alpha-cyano-3-ethoxy-4-hydroxy-5-phenylthiomethylcinnamamid (ST638), reduced I(crom) in a concentration-dependent manner. Daidzein, which does not inhibit tyrosine kinase, was about 10 times less effective than genistein. Herbimycin A had no effect on I(crom). Internal application of these inhibitors from the pipette did not affect I(crom). In conclusion, cromakalim is a potent activator of the ATP-sensitive K(+) channel (K(ATP) channel) in guinea-pig tracheal myocytes. The inhibition of I(crom) by genistein and ST638 may be due to the direct block of the channel from outside.
Assuntos
Broncodilatadores/farmacologia , Cromakalim/farmacologia , Músculo Liso/efeitos dos fármacos , Músculo Liso/fisiologia , Traqueia/efeitos dos fármacos , Traqueia/fisiologia , Trifosfato de Adenosina/fisiologia , Animais , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Cobaias , Masculino , Potenciais da Membrana/efeitos dos fármacos , Músculo Liso/enzimologia , Potássio/farmacologia , Bloqueadores dos Canais de Potássio , Canais de Potássio/fisiologia , Proteínas Tirosina Quinases/antagonistas & inibidores , Traqueia/enzimologiaRESUMO
Expanded polytetrafluoroethylene (ePTFE) can be used as a dura substitute but is associated with leakage of cerebrospinal fluid (CSF) through the suture line. Fibrin glue alone may not prevent this problem. This new method for sealing the suture line in ePTFE membrane uses an absorbable polyglycoic acid mesh soaked with fibrinogen fluid placed on the suture line. Thrombin fluid is then slowly applied to the wet mesh, forming a large fibrin membrane reinforced by the mesh over the suture line. Only one of 33 patients in whom this technique was used had CSF leakage, whereas 12 of 59 patients in whom a dural defect was closed with ePTFE alone showed postoperative subcutaneous CSF collection (p < 0.05). Our clinical experiences clearly show the efficacy of the mesh-and-glue technique to prevent CSF leakage after artificial dural substitution. Mesh and glue can provide an adequate repair for small dural defect. The mesh-and-glue technique may also be used for arachnoid sealing in spinal surgery.
Assuntos
Líquido Cefalorraquidiano , Dura-Máter/cirurgia , Membranas Artificiais , Politetrafluoretileno , Telas Cirúrgicas , Adesivos Teciduais , Biodegradação Ambiental , Craniotomia/métodos , Feminino , Adesivo Tecidual de Fibrina , Fibrinogênio , Humanos , Japão , Masculino , Politetrafluoretileno/efeitos adversos , Técnicas de SuturaRESUMO
To investigate the diversity of morphology in adult T-cell leukaemia/lymphoma (ATL) and its possible association with the pathophysiology of ATL, we selected 36 acute cases and 14 chronic cases phenotypically confirmed to have >90% ATL cells in peripheral blood mononuclear cells. Prototype ATL cells were observed in all cases, although the percentage of all lymphoid cells varied considerably (48.9 +/- 23.8 in acute type, 29.6 +/- 18.9 in chronic type; P = 0.015). Chronic lymphocytic leukaemia (CLL)-like morphology with round nuclei was more frequent in chronic type than in acute type (52.0 +/- 24.9% v 16.6 +/- 13.1%; P < 0.0001). Unusual morphology (UM; lymphoblastic, vacuolated, granular pleomorphic or large cells) was more frequent in acute type than in chronic type (20.1 +/- 18.7% v 2.7 +/- 3.2%; P < 0.0001). Furthermore, there were significant negative and positive correlations of % CLL-like cells and % UM cells respectively, with serum LDH level, hypercalcaemia, performance status, and total number of involved lesions. Cases with aberrant immunophenotype (n = 6) or defective HTLV-1 integration (n = 22) showed lower % CLL-like cells and higher % UM cells than other cases, respectively. Cases with >50% CLL-like cells (n = 7; all chronic type) were younger (53.1 +/- 12.2 v 66.9 +/- 10.6 years; P = 0.038) and showed longer acute-crisis free survival (mean: 16.7 v 3. 0 years; P = 0.012) than chronic cases with <50% CLL-like cells. These results suggest that diversity in genotype, phenotype, morphology and behaviour of ATL are closely associated, and that CLL-like morphology is a good prognostic factor for chronic type.
Assuntos
Leucemia-Linfoma de Células T do Adulto/patologia , Adulto , Tamanho Celular , Genótipo , Vírus Linfotrópico T Tipo 1 Humano/genética , Humanos , Imunofenotipagem , Leucemia-Linfoma de Células T do Adulto/genética , Leucemia-Linfoma de Células T do Adulto/imunologia , PrognósticoRESUMO
We investigated the expression of Fas antigen (CD95) in the pure erythroid cell line AS-E2 in the presence and absence of interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). TNF-alpha induced apoptosis in AS-E2 cells, whereas IFN-gamma did not. In culture containing no IFN-gamma or TNF-alpha, AS-E2 cells expressed little Fas antigen. However, IFN-gamma and IFN-gamma and TNF-alpha both induced expression of Fas antigen and its mRNA within 24 hours after the stimulation. When anti-Fas monoclonal antibody (IgM) was added to AS-E2 cells after the induction of Fas expression, AS-E2 cells underwent apoptosis as shown by the induction of DNA fragmentation. This apoptotic change was inhibited by an inhibitor of caspase-3-like proteases (Ac-DEVD-CHO) and an inhibitor of CED-3/ICE family proteases (Z-Asp-CH2-DCB) but not by an inhibitor of caspase-1-like proteases (Ac-YVAD-CHO), suggesting a role for caspase-3-like proteases in Fas-receptor signaling. Although AS-E2 cells expressed Fas ligand mRNA, treatment with ZB4, an antibody that inhibits Fas-mediated cell death, failed to suppress IFN-gamma- or TNF-alpha-mediated cytotoxicity. These findings suggest that the late erythroid progenitor cells are negatively regulated by IFN-gamma and TNF-alpha, both of which are capable of inducing functional Fas expression.
Assuntos
Apoptose/efeitos dos fármacos , Células Precursoras Eritroides/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Interferon gama/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Receptor fas/biossíntese , Anticorpos Monoclonais/farmacologia , Apoptose/fisiologia , Ácido Aspártico/análogos & derivados , Ácido Aspártico/farmacologia , Caspase 3 , Inibidores de Caspase , Caspases/biossíntese , Caspases/fisiologia , Divisão Celular/efeitos dos fármacos , Fragmentação do DNA , Inibidores Enzimáticos/farmacologia , Células Precursoras Eritroides/citologia , Células Precursoras Eritroides/metabolismo , Proteína Ligante Fas , Humanos , Glicoproteínas de Membrana/biossíntese , Glicoproteínas de Membrana/genética , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética , Oligopeptídeos/farmacologia , RNA Mensageiro/biossíntese , RNA Neoplásico/biossíntese , Proteínas Recombinantes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Células Tumorais Cultivadas , Receptor fas/genética , Receptor fas/fisiologiaRESUMO
The electrophysiological effects of UTP on freshly isolated rat aortic myocytes were examined using the perforated patch clamp technique. Application of alpha,beta-methylene ATP (alphabeta-meATP) and UTP, putative P2X and P2Y2 or P2Y4 purinoceptor agonists, induced transient and oscillatory inward currents, respectively. Experiments with Cl- channel blockers and different external Cl- concentrations demonstrated that the oscillatory current elicited by UTP is attributable to activation of Cl- channels. The transient component elicited by (alphabeta-meATP appeared to be responsible for a non-selective cationic current. With internal application of low-molecular-weight heparin, a blocker of inositol 1,4,5-trisphosphate (InsP3), the oscillatory current elicited by UTP was abolished. The oscillatory current was activated in an all-or-none manner by UTP over the concentration range 0.1 and 1 microM and the frequency and amplitude were independent of the UTP concentration. Under current-clamp mode, UTP produced an oscillatory membrane potential. These results show that rat aortic myocytes have at least two types of P2 receptors. Activation of the P2Y receptor by UTP produces InsP3, which releases Ca2+ from the store site. The resulting increase in intracellular Ca2+ concentration causes the oscillatory Cl- current and the subsequent membrane potential changes.
Assuntos
Membrana Celular/fisiologia , Canais Iônicos/metabolismo , Miocárdio/metabolismo , Uridina Trifosfato/farmacologia , Trifosfato de Adenosina/análogos & derivados , Trifosfato de Adenosina/farmacologia , Animais , Aorta , Quelantes/farmacologia , Canais de Cloreto/antagonistas & inibidores , Canais de Cloreto/metabolismo , Cloretos/metabolismo , Ácido Egtázico/farmacologia , Heparina/farmacologia , Técnicas In Vitro , Canais Iônicos/antagonistas & inibidores , Masculino , Potenciais da Membrana/efeitos dos fármacos , Ratos , Ratos Wistar , Receptores Purinérgicos P2/metabolismo , Vasoconstrição/efeitos dos fármacosRESUMO
Effects of isoproterenol (Iso) and vasoactive intestinal polypeptide (VIP) on voltage-dependent Ca2+ channel (ICa) and Ca(2+)-activated K+ channel current (IK-Ca) in porcine tracheal smooth muscle cells were examined. When K+ currents were inhibited using a Cs-rich pipette solution, application of 0.1-1 microM Iso or 1-10 nM VIP increased ICa by 20-30%. On the other hand, IK-Ca elicited upon depolarization and spontaneous transient outward K+ currents (STOCs) recorded at a holding potential of -50 mV were enhanced by 80-100% in the presence of 0.1 microM Iso or 1 nM VIP.