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1.
Int Endod J ; 52(5): 639-645, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30447154

RESUMO

AIM: To investigate the proliferation and mineralization of a human cementoblast cell line under alkaline conditions. METHODOLOGY: A human cementoblast cell line was cultured in alkaline media with several pHs (pH 7.6, 8.0 and 8.4) without CO2 . Cell numbers, phospho-p44/42 expression, alkaline phosphatase (ALP) activity and mineralization were evaluated. The significance of differences between groups was assessed using two-way analysis of variance 15 (ANOVA) followed by Bonferroni's multiple comparison test (α = 0.01). RESULTS: Cell numbers increased in a time-dependent manner in the high pH medium groups. Western blot analysis revealed the upregulated expression of phospho-p44/42 under alkaline conditions. ALP activity was also increased at pH 8.0 and 8.4. Alizarin red staining revealed increased mineralization in the high pH medium groups. The incorporation of the transient receptor potential ankyrin subfamily member 1 (TRPA1) antagonist HC030031 markedly negated the effect on proliferation and mineralization. CONCLUSIONS: Extracellular alkaline conditions promoted the proliferation and mineralization of human cementoblasts in vitro via TRPA1.


Assuntos
Fosfatase Alcalina , Cemento Dentário , Calcificação Fisiológica , Contagem de Células , Diferenciação Celular , Linhagem Celular , Proliferação de Células , Células Cultivadas , Humanos
2.
Br J Surg ; 103(7): 845-54, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26953091

RESUMO

BACKGROUND: Tumour-infiltrating lymphocytes (TILs) can be used to monitor the immune response, and are important in predicting treatment responses and outcomes for various types of cancer. Recently, specific TIL subsets have been reported to be clinically useful in predicting treatment responses. The CD8+/FOXP3+ TIL ratio (CFR) may be a more sensitive indicator for monitoring immune function. This study investigated the clinical significance and value of CFR as a biomarker to predict treatment responses to neoadjuvant chemotherapy for breast cancer. METHODS: Patients with resectable early-stage breast cancer treated with neoadjuvant chemotherapy at Osaka City University Hospital, Japan, between 2007 and 2013 were included. Oestrogen receptor, progesterone receptor, human epidermal growth factor receptor (HER) 2, Ki-67, CD8 and FOXP3 status were assessed by immunohistochemistry, and correlated with pathological complete response (pCR). RESULTS: A total of 177 patients were included, of whom 90 had a high CFR and 87 a low CFR. Triple-negative breast cancer (TNBC) was more common in the high-CFR group than in the low-CFR group (46 versus 23 per cent; P = 0·002), as was HER2-enriched breast cancer (HER2BC) (27 versus 14 per cent; P = 0·033). Among these patients, the pCR rate was significantly higher in the high-CFR group than in the low-CFR group (TNBC: P = 0·022; HER2BC: P < 0·001). In multivariable analysis high-CFR status was an independent predictor of a favourable prognosis: hazard ratio 0·24 (95 per cent c.i. 0·05 to 0·72; P = 0·015) for TNBC and 0·10 (0·10 to 0·90; P = 0·041) for HER2BC. CONCLUSION: The CFR may be a useful biomarker to predict treatment response to neoadjuvant therapy in aggressive breast cancer subtypes, such as TNBC and HER2BC.


Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/terapia , Linfócitos T CD8-Positivos/metabolismo , Fatores de Transcrição Forkhead/metabolismo , Terapia Neoadjuvante , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/metabolismo , Carcinoma Ductal de Mama/patologia , Carcinoma Ductal de Mama/terapia , Quimioterapia Adjuvante , Feminino , Humanos , Imuno-Histoquímica , Antígeno Ki-67/metabolismo , Mastectomia , Mastectomia Segmentar , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Receptor ErbB-2/metabolismo , Receptores de Estrogênio/metabolismo , Receptores de Progesterona/metabolismo , Neoplasias de Mama Triplo Negativas/metabolismo , Neoplasias de Mama Triplo Negativas/patologia , Neoplasias de Mama Triplo Negativas/terapia
3.
Am J Transplant ; 15(3): 618-27, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25693473

RESUMO

Islet transplantation represents a potentially curative approach for individuals with Type I Diabetes. The requirement for systemic immune suppression to control immune-mediated rejection of transplanted islets and the limited human islet supply represent significant roadblocks to progress for this approach. Islet microencapsulation in alginate offers limited protection in the absence of systemic immunosuppression, but does not support long-term islet survival. The chemokine, CXCL12, can repel effector T cells while recruiting immune-suppressive regulatory T cells (Tregs) to an anatomic site while providing a prosurvival signal for beta-cells. We proposed that coating or encapsulating donor islets with CXCL12 would induce local immune-isolation and protect and support the function of an allo- or xenograft without systemic immune suppression. This study investigated the effect of alginate microcapsules incorporating CXCL12 on islet function. Islet transplantation was performed in murine models of insulin-dependent diabetes. Coating of islets with CXCL12 or microencapsulation of islets with alginate incorporating the chemokine, resulted in long-term allo- and xenoislet survival and function, as well as a selective increase in intragraft Tregs. These data support the use of CXCL12 as a coating or a component of an alginate encapsulant to induce sustained local immune-isolation for allo- or xenoislet transplantation without systemic immunosuppression.


Assuntos
Alginatos/administração & dosagem , Quimiocina CXCL12/administração & dosagem , Transplante das Ilhotas Pancreáticas/imunologia , Animais , Feminino , Ácido Glucurônico/administração & dosagem , Xenoenxertos , Ácidos Hexurônicos/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Transplante Homólogo
4.
Br J Surg ; 100(4): 490-6, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23319435

RESUMO

BACKGROUND: As patients with basal-like breast cancer (BLBC) have a poor prognosis and there is no specifically tailored therapy, molecular biological characterization of BLBC is necessary. c-Kit is a transmembrane receptor tyrosine kinase known to play important roles in various solid cancers. This study classified BLBCs from patients with breast carcinoma, and addressed the significance of c-Kit expression in these tumours. METHODS: Primary breast tumours were stained with antibodies against oestrogen receptor, progesterone receptor, human epidermal growth factor receptor (HER) 2, epidermal growth factor receptor (EGFR), cytokeratin 5/6 and c-Kit. The association between c-Kit, BLBC and survival was analysed. RESULTS: A total of 667 patients with breast cancer were followed up for a median of 39 (range 6-72) months. Some 190 tumours (28·5 per cent) were classified as triple-negative for breast cancer (negative for oestrogen receptor, progesterone receptor and HER2) and 149 (78·4 per cent) had characteristics of BLBC (positive for cytokeratin 5/6 and/or EGFR). c-Kit expression was detected in 111 (16·6 per cent) of 667 tumours. c-Kit-positive tumours were more commonly found among patients with BLBC (42 of 149, 28·2 per cent; P < 0·001) and in patients with nodal metastasis (47 of 216, 21·8 per cent; P = 0·014) than in those without. In patients with BLBC, the prognosis was significantly worse in those with c-Kit expression (P < 0·001). Multivariable logistic regression analysis revealed c-Kit as an independent negative prognostic factor for cancer-specific survival in patients with BLBC (hazard ratio 2·29, 95 per cent confidence interval 1·11 to 4·72). CONCLUSION: c-Kit might be a prognostic marker and possible molecular target for therapy in patients with BLBC.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/mortalidade , Carcinoma Basocelular/mortalidade , Carcinoma Ductal de Mama/mortalidade , Proteínas Proto-Oncogênicas c-kit/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Carcinoma Basocelular/metabolismo , Carcinoma Basocelular/patologia , Carcinoma Ductal de Mama/metabolismo , Carcinoma Ductal de Mama/patologia , Feminino , Humanos , Imuno-Histoquímica , Metástase Linfática , Pessoa de Meia-Idade , Prognóstico , Receptor ErbB-2/metabolismo , Receptores de Progesterona/metabolismo , Análise de Sobrevida
5.
Br J Cancer ; 105(7): 996-1001, 2011 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-21863023

RESUMO

BACKGROUND: Myofibroblasts in the cancer microenvironment have recently been implicated in tumour growth and metastasis of gastric cancer. However, the mechanisms responsible for the regulation of myofibroblasts in cancer-associated fibroblasts (CAFs) remain unclear. This study was performed to clarify the mechanisms for regulation of myofibroblasts in gastric cancer microenvironment. METHODS: Two CAFs (CaF-29 and CaF-33) from the tumoural gastric wall and a normal fibroblast (NF-29) from the nontumoural gastric wall, 4 human gastric cancer cell lines from scirrhous gastric cancer (OCUM-2MD3 and OCUM-12), and non-scirrhous gastric cancer (MKN-45 and MKN-74) were used. Immunofluorescence microscopy by triple-immunofluorescence labelling (α-SMA, vimentin, and DAPI) was performed to determine the presence of α-SMA-positive myofibroblasts. Real-time RT-PCR was performed to examine α-SMA mRNA expression. RESULTS: Immunofluorescence microscopy showed that the frequency of myofibroblasts in CaF-29 was greater than that in NF-29. The number of myofibroblasts in gastric fibroblasts gradually decreased with serial passages. Transforming growth factor-ß (TGF-ß) significantly increased the α-SMA expression level of CAFs. Conditioned medium from OCUM-2MD3 or OCUM-12 cells upregulated the α-SMA expression level of CAFs, but that from MKN-45 or MKN-74 cells did not. The α-SMA upregulation effect of conditioned medium from OCUM-2MD3 or OCUM-12 cells was significantly decreased by an anti-TGF-ß antibody or Smad2 siRNA. CONCLUSION: Transforming growth factor-ß from scirrhous gastric carcinoma cells upregulates the number of myofibroblasts in CAFs.


Assuntos
Adenocarcinoma Esquirroso/patologia , Fibroblastos/patologia , Miofibroblastos/patologia , Neoplasias Gástricas/patologia , Fator de Crescimento Transformador beta/metabolismo , Adenocarcinoma Esquirroso/metabolismo , Idoso , Western Blotting , Meios de Cultivo Condicionados/farmacologia , Fibroblastos/metabolismo , Imunofluorescência , Mucosa Gástrica/metabolismo , Humanos , Técnicas Imunoenzimáticas , Masculino , Miofibroblastos/metabolismo , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína Smad2/antagonistas & inibidores , Proteína Smad2/genética , Proteína Smad2/metabolismo , Estômago/patologia , Neoplasias Gástricas/metabolismo , Fator de Crescimento Transformador beta/genética , Células Tumorais Cultivadas , Regulação para Cima
6.
Br J Cancer ; 103(2): 249-55, 2010 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-20551954

RESUMO

PURPOSE: Triple-negative breast cancer (TNBC), a subtype of breast cancer that is oestrogen receptor (ER) negative, progesterone receptor (PR) negative, and human epidermal growth factor receptor 2 (HER2) negative, has a poor prognosis. Although a correlation between E-cadherin expression level and outcome has been demonstrated among all types of breast cancer, little is known about the significance of E-cadherin expression levels in TNBC. METHODS: A total of 574 patients who had undergone a resection of a primary breast cancer except for invasive lobular carcinomas were enrolled in this study. Expressions of ER, PR, HER2, and E-cadherin were assessed by immunohistochemistry. We examined the association between TNBC and other clinicopathological variables and evaluated the significance of the E-cadherin expression. RESULTS: Among the 574 breast cancer cases, 123 (21.4%) revealed a triple-negative phenotype. Patients with TNBC experienced more frequent lymph node metastasis (P=0.024) and a poorer prognosis (P<0.001) in comparison with non-TNBC patients. Triple-negative breast cancer was an independent prognostic factor. Reduced levels of E-cadherin were observed in 238 (41.5%) of the 574 breast cancer cases. E-cadherin reduction was significantly frequent in cases of TNBC (P<0.001) and lymph node metastasis (P=0.032). Furthermore, in the 123 TNBC cases, the prognosis of patients with an E-cadherin-negative expression was significantly worse than that of E-cadherin-positive patients (P=0.0265), especially for those in clinical stage II (P=0.002). A multivariate logistic regression analysis showed a reduction of the E-cadherin expression to be an independent prognostic factor (P=0.046). CONCLUSION: E-cadherin expression may be a useful prognostic marker for classifying subgroups of TNBC.


Assuntos
Neoplasias da Mama/metabolismo , Caderinas/metabolismo , Receptores de Estrogênio/análise , Neoplasias da Mama/mortalidade , Neoplasias da Mama/patologia , Receptores ErbB/análise , Humanos , Pessoa de Meia-Idade , Prognóstico , Receptores de Progesterona/análise
7.
Br J Cancer ; 102(5): 898-907, 2010 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-20145613

RESUMO

BACKGROUND: Many kinds of solid tumour have heterogeneously a hypoxic environment. Tumour hypoxia reported to be associated with more aggressive tumour phenotypes such as high metastatic ability and resistance to various anti-cancer therapies which may lead to a poorer prognosis. However, the mechanisms by which hypoxia affects the aggressive phenotypes remain unclear. METHODS: We established a scirrhous gastric carcinoma cell line (OCUM-12) from ascites associated with scirrhous gastric carcinoma, and a hypoxia-resistant cancer cell line (OCUM-12/Hypo) was cloned from OCUM-12 cells by continuous exposure to 1% oxygen. RESULTS: Histologic findings from orthotopic tumours derived from parent OCUM-12 cells and daughter OCUM-12/Hypo cells revealed poorly differentiated adenocarcinoma with extensive fibrosis that resembled human scirrhous gastric cancer. Necrotic lesions were frequently detected in the OCUM-12 tumours but were rarely found in the OCUM-12/Hypo tumours, although both types had multiple hypoxic loci. Apoptosis rate of OCUM-12 cells was increased to 24.7% at 1% O(2), whereas that of OCUM-12/Hypo was 5.6%. The OCUM-12/Hypo orthotopic models developed multiple metastases to the peritoneum and lymph nodes, but the OCUM-12 models did not. OCUM-12/Hypo cells showed epithelial-to-mesenchymal transition and high migratory and invasive activities in comparison with OCUM-12 cells. The mRNA expression levels of both E-cadherin and zonula occludens ZO-1 and ZO-2 decreased in OCUM-12/Hypo cells, and that of vimentin, Snail-1, Slug/Snail-2, Twist, ZEB-1, ZEB-2, matrix metalloproteinase-1 (MMP-1), and MMP-2 were increased in OCUM-12/Hypo cells. CONCLUSION: OCUM-12 and OCUM-12/Hypo may be useful for the elucidation of disease progression associated with scirrhous gastric cancer in the setting of chronic hypoxia.


Assuntos
Adenocarcinoma Esquirroso/patologia , Hipóxia , Neoplasias Peritoneais/secundário , Neoplasias Gástricas/patologia , Adenocarcinoma Esquirroso/genética , Adenocarcinoma Esquirroso/metabolismo , Animais , Apoptose , Western Blotting , Adesão Celular , Linhagem Celular Tumoral , Movimento Celular , Mapeamento Cromossômico , Citometria de Fluxo , Humanos , Técnicas Imunoenzimáticas , Cariotipagem , Perda de Heterozigosidade , Masculino , Metaloproteinases da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Neoplasias Peritoneais/genética , Neoplasias Peritoneais/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo
8.
BJS Open ; 3(1): 38-47, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30734014

RESUMO

Background: Recently, evaluation of quality of life (QOL) has been recognized as a significant outcome measure in the treatment of several cancers. In this study, the Anti-Cancer Drugs-Breast (ACD-B) QOL score was used to assess disease-specific survival in women with breast cancer undergoing preoperative chemotherapy (POC). Methods: QOL-ACD-B scores were evaluated before and after POC. The cut-off value of QOL-ACD-B contributing to events such as relapse or death was calculated by receiver operating characteristic (ROC) curve analysis. Results: In 300 women with breast cancer treated with POC, QOL was significantly reduced (P < 0·001). A high QOL-ACD-B score before POC was an independent factor in the multivariable analysis of overall survival (hazard ratio 0·26, 95 per cent c.i. 0·04 to 0·96). Conclusion: Evaluation by QOL-ACD-B before POC may be useful to predict the prognosis of patients with breast cancer undergoing POC.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Qualidade de Vida , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/patologia , Neoplasias da Mama/reabilitação , Neoplasias da Mama/cirurgia , Quimioterapia Adjuvante , Feminino , Humanos , Japão , Estimativa de Kaplan-Meier , Metástase Linfática , Mastectomia , Pessoa de Meia-Idade , Terapia Neoadjuvante , Prognóstico , Psicometria , Curva ROC , Resultado do Tratamento
10.
J Clin Invest ; 96(5): 2431-7, 1995 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-7593631

RESUMO

Heme oxygenase is a heme-oxidizing enzyme which generates biliverdin and carbon monoxide (CO). The present study was designed to elucidate whether CO endogenously produced by this enzyme serves as an active vasorelaxant in the hepatic microcirculation. Microvasculature of the isolated perfused rat liver was visualized by dual-color digital microfluorography to alternately monitor sinusoidal lining and fat-storing Ito cells. In the control liver, the CO flux in the venous effluent ranged at 0.7 nmol/min per gram of liver. Administration of a heme oxygenase inhibitor zinc protoporphyrin IX (1 microM) eliminated the baseline CO generation, and the vascular resistance exhibited a 30% elevation concurrent with discrete patterns of constriction in sinusoids and reduction of the sinusoidal perfusion velocity. The major sites of the constriction corresponded to local sinusoidal segments colocalized with Ito cell which were identified by imaging their vitamin A autofluorescence. The increase in the vascular resistance and sinusoidal constriction were attenuated significantly by adding CO (1 microM) or a cGMP analogue 8-bromo-cGMP (1 microM) in the perfusate. From these findings, we propose that CO can function as an endogenous modulator of hepatic sinusoidal perfusion through a relaxing mechanism involving Ito cells.


Assuntos
Monóxido de Carbono/metabolismo , Fígado/irrigação sanguínea , Animais , Inibidores Enzimáticos/farmacologia , Heme Oxigenase (Desciclizante)/antagonistas & inibidores , Heme Oxigenase (Desciclizante)/fisiologia , Fígado/fisiologia , Masculino , Microcirculação/fisiologia , Perfusão , Protoporfirinas/farmacologia , Ratos , Ratos Wistar , Vasodilatação/efeitos dos fármacos
11.
J Clin Invest ; 96(3): 1202-7, 1995 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-7657792

RESUMO

Homozygous adhalin gene mutations were found in three patients from two consanguineous families with autosomal recessive childhood onset muscular dystrophy. Muscle biopsies from patients in each family showed complete absence of adhalin. Sequencing of adhalin cDNA prepared from skeletal muscle by reverse transcription PCR demonstrated a cytosine to thymidine substitution at nt 229 in the patient in family 1 and an adenine to guanine substitution at nt 410 and a 15-base insertion between nt 408 and 409 in the two patients in family 2. Sequencing of genomic DNA prepared from peripheral blood leukocytes by PCR confirmed these mutations. The parents in each family were found to be heterozygous for the respective mutations. These adhalin gene mutations are presumed to be responsible for the absence of adhalin in the skeletal muscle. Adhalin deficiency likely causes disruption of the muscle cell membrane, resulting in dystrophic changes in the skeletal muscle similar to dystrophin deficiency in Duchenne muscular dystrophy.


Assuntos
Proteínas do Citoesqueleto/deficiência , Proteínas do Citoesqueleto/genética , Genes Recessivos , Glicoproteínas de Membrana/deficiência , Glicoproteínas de Membrana/genética , Músculo Esquelético/metabolismo , Distrofias Musculares/genética , Mutação , Adulto , Idade de Início , Sequência de Aminoácidos , Sequência de Bases , Biópsia , Criança , Consanguinidade , Primers do DNA , Elementos de DNA Transponíveis , DNA Complementar , Feminino , Homozigoto , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Músculo Esquelético/patologia , Distrofias Musculares/patologia , Linhagem , Reação em Cadeia da Polimerase/métodos , Valores de Referência , Sarcoglicanas
12.
J Natl Cancer Inst ; 88(11): 742-6, 1996 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-8637028

RESUMO

BACKGROUND: Although hepatitis C virus (HCV) infection is recognized as an important risk factor for hepatocellular carcinoma HCC), the strength of this association has been inconsistent. In addition, the role of specific HCV genotypes in HCC progression has not yet been determined. PURPOSE: We conducted a case-control study to estimate the relative risk (RR) of HCC in relation to HCV infection among residents of the Fukuoka Prefecture, where HCC risk is among the highest in Japan, and to examine whether the risk differs according to HCV genotypes and/or HCV RNA titers. METHODS: Stored serum samples obtained from 91 patients with HCC and 410 healthy control subjects, who had been frequency matched to the patients with regard to sex and age, were tested for antibodies to HCV by use of second-generation immunoradiometric and immunoblot assays. The presence of serum HCV RNA and of specific HCV genotypes was determined by use of polymerase chain reaction-based assays, and HCV RNA titers were measured by use of a branched DNA assay. RESULTS: Antibodies to HCV were detected in 71 patients (78.0%) and in 30 control subjects (7.3%), of whom 57 patients and 25 control subjects had serum HCV RNA. One patient was positive for HCV RNA but not for antibodies to HCV. The sex- and age-adjusted RR of HCC among individuals positive for antibodies to HCV was estimated to be 53.7 (95% confidence interval [CI] = 27.1-106.2). Antibodies to HCV were much more prevalent among patients negative for serum, hepatitis B surface antigen (HBsAg) (69 of 72, 95.8 %) than among HBsAg-positive patients (two of 19, 10.5%); the RR increased to 339.6 (95% CI = 96.5-1195.8) in the separate analysis of HBsAg-negative subjects. The most frequent genotype among HCV RNA-positive subjects was type lb (also called type II) (found in 49 [86.0%] of 57 patients and in 15 [60.0%] of 25 control subjects); individuals with type 1b infection experienced a significantly elevated risk (RR = 3.8; 95% CI = 1.0-13.9) compared with the risk observed for individuals with type 2a (also called type III) infection. No statistically significant association between HCV RNA titers and HCC was evident. CONCLUSIONS: HCV infection, particularly type 1b infection, plays an important role in the development of HCC among the study population. We estimated that approximately 78% (95% CI = 69%-86%) of the HCCs that occur in this high-risk area are attributable to HCV infection, if we assume that the patients in this study were representative population samples. IMPLICATIONS: Further studies are needed to clarify potential risk factors, including specific HCV genotypes, for progression to HCC among HCV carriers.


Assuntos
Carcinoma Hepatocelular/etiologia , Hepatite C/complicações , Neoplasias Hepáticas/etiologia , Adulto , Idoso , Feminino , Hepacivirus/genética , Antígenos de Superfície da Hepatite B/análise , Anticorpos Anti-Hepatite C/sangue , Humanos , Masculino , Pessoa de Meia-Idade , RNA Viral/sangue , Fatores de Risco
13.
Biochim Biophys Acta ; 1237(2): 189-92, 1995 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-7632713

RESUMO

A cyanobacterium, Synechococcus species PCC7942, has a gene encoding a copper-transporting P-type ATPase, which is located in the thylakoid membrane. At the 5'-upstream of this ATPase gene, we identified another gene, which was supposed to be implicated in a copper-transport process. This novel gene was found to encode a putative pore-forming membrane protein that belongs to a growing family of homologous intrinsic membrane proteins (the MIP family of proteins), which include the major intrinsic protein (MIP) from animal lens fibre junction membranes, the tonoplast intrinsic protein (TIP) from vacuolar membranes of higher plants, and the Escherichia coli glycerol facilitator (GlpF) in the cytoplasmic membrane. The deduced product, named SmpX (Synechococcus membrane protein), is highly homologous throughout its entire sequence to these intrinsic membrane proteins which were postulated to be pore-forming proteins involved in a variety of transport processes. The primary amino acid sequence of SmpX shares all properties characteristic for members of the MIP family. SmpX is more similar to the eukaryotic members (e.g., nodulin-26 from soybean) than to the prokaryotic ones.


Assuntos
Cianobactérias/genética , Proteínas Musculares , Porinas/genética , Sequência de Aminoácidos , Sequência de Bases , DNA Bacteriano , Dados de Sequência Molecular , Homologia de Sequência de Aminoácidos
14.
Biochim Biophys Acta ; 1326(1): 1-6, 1997 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-9188794

RESUMO

Metal-transporting P-type ATPases were recently proposed to constitute a newly emerged sub-family of cation-transporting P-type ATPases, and are known to occur widely in prokaryotes and eukaryotes. However, no instance has been reported for higher plants. A cDNA clone encoding a metal-transporting P-type ATPase was thus searched for, if present, and was identified in Arabidopsis thaliana. The amino acid sequence, predicted from the determined nucleotide sequence for the cloned cDNA, shows all the critical features common to known metal-transporting P-type ATPases. This plant P-type ATPase has a typical metal-binding motif at its N-terminal portion. The newly isolated Arabidopsis gene, named PAA1, provides us with the first instance of putative metal-transporting P-type ATPases in higher plants. Some results of genomic analyses for this gene are also presented.


Assuntos
Adenosina Trifosfatases/genética , Arabidopsis/enzimologia , DNA Complementar/genética , Genes de Plantas , Sequência de Aminoácidos , Arabidopsis/genética , Sítios de Ligação , Clonagem Molecular , Dados de Sequência Molecular , Alinhamento de Sequência
15.
Diabetes ; 47(1): 104-12, 1998 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9421382

RESUMO

[Ca2+]i and whole-cell membrane current were measured in microvascular endothelial cells from bovine brain. The effects of histamine on [Ca2+]i were examined, and the acute effect of changing extracellular glucose concentration on Ca2+ homeostasis was investigated. Application of 10 micromol/l histamine evoked an initially transient and then sustained increase in [Ca2+]i in normal Krebs solution, but only the transient component in Ca2+-free solution, thereby indicating that histamine mobilizes Ca2+ both from intracellular store sites and extracellular space. The effects of histamine on [Ca2+]i were inhibited by the H2 antagonists, ranitidine and cimetidine, but not by the H1 antagonist, pyrilamine. Incubation of the cells for 2 h in solutions containing low (1.1 and 2.3 mmol/l) or high (23 mmol/l) concentrations of glucose did not influence the resting level of [Ca2+]i. Treatment with low concentrations of glucose did not impair histamine-induced Ca2+ mobilization. On the other hand, when histamine was applied to the cells pretreated with 23 mmol/l glucose, it failed to mobilize Ca2+ from both intracellular store sites and extracellular space. The effect of histamine was mimicked by dibutyryl cyclic AMP, but glucose overload failed to inhibit this, suggesting that glucose overload inhibits H2 receptor-mediated cyclic AMP production. Glucose overload-induced impairment of histamine action was reversed by pretreatment with staurosporine and calphostin C and mimicked by phorbol-12,13-dibutyrate, thereby suggesting the involvement of protein kinase C in the high glucose-induced inhibition of Ca2+ mobilization. Whole-cell membrane current measurement showed that there was no difference in the membrane currents between control and high glucose-treated cells. These results indicate that in bovine brain microvascular endothelial cells, histamine induces Ca2+ release from intracellular store sites and subsequent entry from the extracellular space through the activation of H2 receptors. Glucose overload acutely inhibits histamine-induced Ca2+ mobilization by the activation of protein kinase C.


Assuntos
Encéfalo/irrigação sanguínea , Cálcio/metabolismo , Endotélio Vascular/metabolismo , Glucose/farmacologia , Receptores Histamínicos H2/fisiologia , Animais , Bradicinina/farmacologia , Bucladesina/farmacologia , Bovinos , Células Cultivadas , Cimetidina/farmacologia , AMP Cíclico/farmacologia , Relação Dose-Resposta a Droga , Endotélio Vascular/química , Endotélio Vascular/citologia , Histamina/farmacologia , Antagonistas dos Receptores Histamínicos/farmacologia , Homeostase , Microcirculação , Neomicina/farmacologia , Neurotransmissores/farmacologia , Proteína Quinase C/fisiologia , Pirilamina/farmacologia , Ranitidina/farmacologia , Receptores Histamínicos H2/análise , Estaurosporina/farmacologia
16.
Rev Med Virol ; 8(2): 97-111, 1998 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10398498

RESUMO

Intradermal administration maybe useful in lowering the cost of vaccination against hepatitis B significantly, and may also be helpful for the rapid induction of antibodies, reversing non-responsiveness, improving postexposure prophylaxis and immunising immunocompromised people. In addition, delayed type hypersensitivity skin reaction to the vaccine could serve as a useful marker for the acquisition of T helper type 1 immunoreactivity in vivo. However, there are some disadvantages when using intradermal vaccinations, including the requirement for skilful administration, the absence of approval from licensing authorities, the development of local skin reactions and a lower antibody response when 1/10 of the standard vaccine dose is used. This requires that appropriate vaccination regimens, including the correct vaccine dosage, and vaccination schedule are followed. In the future, a similar vaccination strategy might also be applied for the prevention and control of other infectious diseases. Copyright 1998 John Wiley & Sons, Ltd.

17.
Arch Intern Med ; 158(2): 177-81, 1998 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-9448556

RESUMO

BACKGROUND: Interferon alfa is used widely for patients with chronic hepatitis C virus (HCV) infection. Little is known, however, of the relationship between patients' sex and the effectiveness of interferon alfa treatment in these patients. METHODS: We treated 311 patients (199 men and 112 women) with human lymphoblastoid interferon (6 million units subcutaneously every day for 2 weeks and 3 times a week for 22 weeks) and observed them for an additional 6 months. Serum HCV RNA levels and genotype were tested by polymerase chain reaction before treatment. A liver biopsy was also done. For the purposes of this study, a complete response was defined as the elimination of HCV RNA for at least 6 months after the termination of treatment. RESULTS: The rate of complete response was 27.1% for men and 24.1% for women. With multiple logistic regression analysis, the HCV RNA level (P < .001), genotype (P < .001), patients' sex (P < .05), and the interaction between sex and age were associated with a complete response to interferon alfa. The rate of complete response was 33.3% in men aged 39 years and younger, 25.0% in men aged 40 years and older. 75.0% in women aged 39 years and younger, and 15.6% in women aged 40 years and older. The odds ratio by group was 1.00, 0.72, 4.38, and 0.21, respectively. CONCLUSIONS: Our finding that women aged 39 years and younger are responsive to interferon alfa treatment suggests that hormonal activity, in particular the level of estrogen, may be associated with the sustained elimination of HCV.


Assuntos
Antivirais/uso terapêutico , Hepacivirus/efeitos dos fármacos , Hepatite C Crônica/tratamento farmacológico , Interferon-alfa/uso terapêutico , Adulto , Fatores Etários , Feminino , Hepacivirus/genética , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , RNA Viral/análise , Fatores Sexuais , Resultado do Tratamento
18.
Acta Neurol Scand Suppl ; 166: 51-3, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-8686442

RESUMO

The historical background and current status of the inhalation protocols for the xenon CT CBF method are reviewed. A wash-in method has been established and widely accepted as a standard protocol. A wash-in/washout method is a useful alternative for a practical CBF study. The effect of flow activation with xenon gas can be minimized by shortening the inhalation time. Xe CT CBF method remains to be an indispensable technology for the clinical decision-making in various brain disorders.


Assuntos
Encéfalo/irrigação sanguínea , Meios de Contraste , Tomografia Computadorizada por Raios X/instrumentação , Radioisótopos de Xenônio , Xenônio , Velocidade do Fluxo Sanguíneo/efeitos dos fármacos , Velocidade do Fluxo Sanguíneo/fisiologia , Humanos , Variações Dependentes do Observador , Valores de Referência , Fluxo Sanguíneo Regional/fisiologia
19.
Acta Neurol Scand Suppl ; 166: 139-43, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-8686431

RESUMO

Cerebral blood flow (CBF) is usually decreased in patients with hypertensive putaminal hemorrhage (HPH). However, there are few reports concerning cerebrovascular reserve capacity (CRC) in these cases. This study evaluated cerebral hemodynamics in patients with HPH by measuring CBF and CRC. CBF and CRC were measured by stable xenon enhanced computed tomography (Xe-CT) in 11 patients with HPH (HPH group) and 11 patients with essential hypertension without intracerebral hematoma (non-HPH group). CBFs of the hemisphere and thalamus in the HPH group were lower than those in the non-HPH group. And the CBF of the hemisphere was increased transiently after the surgical evacuation of the hematoma. Thereafter, it fell gradually. The CRCs were also lower in acute stage of the HPH group. The CRC recovered during the chronic stage. Hemodynamics in patients with HPH can be modulated by surgical removal of hematoma. However, some adjunct therapies are necessary to prevent delayed neuronal inactivity, Stable Xe-CT with acetazolamide test is useful to evaluate hemodynamics in the HPH patients.


Assuntos
Acetazolamida , Hemorragia Cerebral/fisiopatologia , Meios de Contraste , Hemodinâmica/fisiologia , Hipertensão/complicações , Putamen/irrigação sanguínea , Tomografia Computadorizada por Raios X , Xenônio , Adulto , Idoso , Velocidade do Fluxo Sanguíneo/fisiologia , Hemorragia Cerebral/cirurgia , Dominância Cerebral/fisiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/fisiopatologia , Fluxo Sanguíneo Regional/fisiologia , Tálamo/irrigação sanguínea
20.
Acta Neurol Scand Suppl ; 166: 6-9, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-8686445

RESUMO

We compared stable xenon enhanced X-ray computed tomography (Xe-CT) with Technetium-99m ethylsteinate dimer single-photon emission computed tomography (ECD-SPECT) in 12 patients. We evaluated the cerebral blood flow (CBF) values in the territory of the anterior, middle and posterior cerebral artery, and in the thalamus. The CBF values were higher in ECD-SPECT than in Xe-CT except for the values of the thalamus. The posterior cerebral artery territory showed a lower correlation and the thalamus had no correlation between two methods. We discussed causes of these differences.


Assuntos
Encéfalo/irrigação sanguínea , Hemorragia Cerebral/diagnóstico por imagem , Infarto Cerebral/diagnóstico por imagem , Tomografia Computadorizada de Emissão de Fóton Único/métodos , Tomografia Computadorizada por Raios X , Isótopos de Xenônio , Idoso , Idoso de 80 Anos ou mais , Velocidade do Fluxo Sanguíneo/fisiologia , Hemorragia Cerebral/fisiopatologia , Infarto Cerebral/fisiopatologia , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Pessoa de Meia-Idade , Fluxo Sanguíneo Regional/fisiologia , Tecnécio , Tálamo/irrigação sanguínea
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