Assuntos
Transtornos de Deficit da Atenção e do Comportamento Disruptivo/sangue , Transtornos de Deficit da Atenção e do Comportamento Disruptivo/etiologia , Glicemia/fisiologia , Insulinoma/complicações , Eletroencefalografia , Feminino , Humanos , Estudos Longitudinais , Pessoa de Meia-Idade , PolissonografiaRESUMO
The telomere repeat lengths of BL cell lines were quantified by measuring terminal restriction fragment (TRF). Epstein-Barr virus (EBV)-positive Namalwa, Raji, and EB-3 cell lines have long telomeres, i.e. TRFs 10-19 kbp, whereas the Daudi cell line, producing a transformation-defective EBV mutant, has TRFs approximately 2.2 kbp. EBV-negative BJAB and DG75 cell lines have short TRFs 3.9-5.4 kbp, shorter than the approximately 12 kbp TRFs in PBLs. Telomerase activities of these BL cell lines are similar. TRFs of non-BL lymphoma cell lines are 2.3-5.5 kbp. Fluorescent in situ hybridization (FISH) studies of these cell lines showed remarkable heterogeneity of telomere size in chromosomes in the same BL cell. These results suggest that EBV-positive and EBV-negative BL cell lines have experienced various telomere dynamics.
Assuntos
Linfoma de Burkitt/genética , Linfoma de Burkitt/virologia , Herpesvirus Humano 4/isolamento & purificação , Telomerase/metabolismo , Telômero/genética , Sequência de Bases , Linfoma de Burkitt/enzimologia , Linhagem Celular Tumoral , DNA de Neoplasias/genética , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/patogenicidade , Humanos , Hibridização in Situ Fluorescente , Mutação , Telomerase/genética , Telômero/enzimologia , Telômero/ultraestruturaRESUMO
Associations of Epstein-Barr virus (EBV) and autoimmune diseases have been hypothesized. We have analysed IgG antibodies to EBV nuclear antigen (EBNA)-2 in sera from Japanese patients with autoimmune systemic connective tissue diseases (CTD), exemplified by systemic lupus erythematosus (SLE), primary Sjogren's syndrome (SS), rheumatoid arthritis (RA), systemic sclerosis (SSc) and secondary SS (classical CTDs complicated with SS). An enzyme-linked immunosorbent assay (ELISA) which uses glutathione-S-transferase polypeptides fused to EBV nuclear antigen (EBNA)-2 and EBNA-1 was developed. Ratios of IgG antibody reactivity to whole IgG concentrations of sera were calculated to normalize EBNA-2 and EBNA-1 antibody levels to the hypergammaglobulinaemia that occurs in CTD. The ELISA optical density OD(450) readings of IgG antibodies to both the amino-terminal aa 1-116 of EBNA-2 and carboxyl-terminal aa 451-641 of EBNA-1 were elevated significantly in patients with SLE, primary SS, RA, SSc and secondary SS when compared to EBNA-1. The OD readings were divided by serum IgG concentrations to normalize for the hypergammaglobulinaemia. The specific levels of IgG antibodies to the amino-terminal region of EBNA-2 were elevated in patients with SLE, primary SS or RA, as well as those with secondary SS complicated with SLE or RA. The EBNA-2 amino-terminal region contains a polyproline tract and a proline-rich sequence and has considerable amino acid sequence homology with many cellular proline-rich proteins. High ratios of EBNA-2 aa 1-116 to EBNA-1 aa 451-641 IgG antibody levels which probably suggest reactivation of EBV latent infection were associated significantly with pulmonary involvement in SS patients. These results are consistent with the hypothesis that the sequence similarity between the amino-terminal region of EBNA-2 and proline-rich cellular proteins is associated with pathogenesis in a subpopulation of CTD patients, possibly by the molecular mimicry-epitope shift mechanism.
Assuntos
Anticorpos Antivirais/sangue , Doenças do Tecido Conjuntivo/imunologia , Antígenos Nucleares do Vírus Epstein-Barr/imunologia , Imunoglobulina G/sangue , Pulmão/imunologia , Adulto , Artrite Reumatoide/imunologia , Artrite Reumatoide/virologia , Estudos de Casos e Controles , Doenças do Tecido Conjuntivo/virologia , Ensaio de Imunoadsorção Enzimática/métodos , Infecções por Vírus Epstein-Barr/imunologia , Humanos , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/virologia , Síndrome de Sjogren/imunologia , Síndrome de Sjogren/virologia , Estatísticas não Paramétricas , Proteínas Virais , Latência ViralRESUMO
We have generated a mouse IgG1 monoclonal antibody (mAb) that recognizes amino acids 1-58 of Epstein-Barr virus (EBV) nuclear antigen 2 (EBNA 2) of type 1 EBV strain B95-8. mAb Y101 also reacted with EBNA 2 of EBV type 2 strains MISP and Jijoye in immunoblots, whereas Jijoye EBNA 2 was not detected by the widely used mAb PE2. mAb Y101, in contrast to mAb PE2, reacted with faster migrated, hypophosphorylated proteins of type 1 EBNA 2 as intensely as slower migrated, hyperphosphorylated ones. mAb Y101 did not react in fixed-cell immunostaining or cell extract immunoprecipitation. The results implicate that the amino-terminal epitope is not exposed in a native form, consistent with the previously reported idea of self-association of EBNA 2 through the amino-terminus. mAb Y101 is the first mAb to the EBNA 2 amino-terminus and will be useful for further analyses of the structure and function of EBNA 2.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/imunologia , Especificidade de Anticorpos , Antígenos Nucleares do Vírus Epstein-Barr/química , Antígenos Nucleares do Vírus Epstein-Barr/imunologia , Sequência de Aminoácidos , Animais , Linhagem Celular , Mapeamento de Epitopos , Antígenos Nucleares do Vírus Epstein-Barr/administração & dosagem , Herpesvirus Humano 4/imunologia , Herpesvirus Humano 4/metabolismo , Imunização , Camundongos , Camundongos Endogâmicos BALB C , Proteínas ViraisRESUMO
We consider that off-pump coronary artery bypass grafting (CABG) [OPCAB], which results in local myocardial ischemia, is more effective for patients with acute myocardial infarction (AMI) than conventional CABG under cardiac arrest with global myocardial ischemia. Twenty-one patients (15 males, 6 females) received OPCAB for AMI, among whom surgery was performed following percutaneous coronary intervention (PCI) failure in 4 and PCI was performed prior to OPCAB in 2, while PCI was not performed in the remaining 15. Preoperatively, 16 patients had intraaortic balloon pumping (IABP), and 4 had IABP and percutaneous cardiopulmonary support (PCPS). The mean interval from onset to surgery was 11.7 (range 3 to 40) hours. In 20 cases, a complete revascularization was performed. The mean number of bypasses was 2.3 and OPCAB was carried out in 14 patients. In 2 cases, OPCAB was converted to on-pump beating CABG for complete revascularization. Fourteen patients (67%), each maintained with preoperative left ventricular ejection fraction (EF), were discharged with an elective bypass. Four patients died after on-pump beating CABG, in whom EF was lower than 10%. In addition, 3 died of low cardiac output syndrome (LOS) under PCPS and 1 of ventricular fibrillation. Based on our results, we considered that complete revascularization using OPCAB was effective for cases of AMI with PCI difficulty. However, in shock cases requiring PCPS, cardiac function was not improved even after revascularization. Therefore, it is necessary to study new procedures for shock cases during the period from onset to surgery.
Assuntos
Ponte de Artéria Coronária sem Circulação Extracorpórea , Infarto do Miocárdio/cirurgia , Idoso , Ponte Cardiopulmonar , Ponte de Artéria Coronária/mortalidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/mortalidade , Prognóstico , Taxa de SobrevidaRESUMO
Epstein-Barr virus nuclear antigen 1 (EBNA-1) is essential for the maintenance of latent EBV plasmids, and is also a transcriptional regulator. Nuclear lamins, components of the nuclear lamina, have also been found in the nucleoplasm. We report here that EBNA-1 coincided with lamin B1 in the nucleoplasm and around the nuclear rim during S-phase by confocal microscopy of cells transfected with EBNA-1 in the absence of EBV plasmids. Lamin B1, which is rarely detected in nuclear soluble fractions, was detected in chromatin and nuclear matrix fractions of the EBNA-1-expressing cells. These observations suggest that EBNA-1 colocalizes with lamin B1 in the subnuclear sites.
Assuntos
Antígenos Nucleares do Vírus Epstein-Barr/metabolismo , Lamina Tipo B/metabolismo , Matriz Nuclear/metabolismo , Animais , Linfoma de Burkitt , Células CHO , Linhagem Celular , Cromatina/metabolismo , Cricetinae , Antígenos Nucleares do Vírus Epstein-Barr/genética , Humanos , Fase S , Transfecção , Células Tumorais CultivadasRESUMO
Activation-induced cell death (AICD) is a well-known mechanism of peripheral T cell tolerance that depends upon an interaction between Fas and Fas ligand (FasL). In this study, we demonstrate that the administration of a soluble form of anti-FasL Ab, FLIM58, results in severe destructive autoimmune exocrinopathy in the murine model of human Sjögren's syndrome (SS), and we found that an organ-specific autoantigen may play an important role on down-modulation of AICD. A high titer of serum autoantibodies against 120-kDa alpha-fodrin autoantigen was detected in the FLIM58-treated mice, and splenic T cell culture supernatants contained high levels of IFN-gamma. In vitro T cell apoptosis assay indicated that FasL-mediated AICD is down-regulated by autoantigen stimulation in spleen cells from the murine SS model, but not from Fas-deficient MRL/lpr mice and FasL-deficient MRL/gld mice. FasL undergo metalloproteinase-mediated proteolytic processing in their extracellular domains, resulting in the release of soluble trimeric ligands (soluble FasL). We showed that the processing of soluble FasL occurs in autoantigen-specific CD4(+) T cells, and that a significant increase in expressions of metalloproteinase-9 mRNA was observed in spleen cells from SS model mice. These findings indicate that the increased generation of soluble FasL inhibits the normal AICD process, leading to the proliferation of effector CD4(+) T cells in the murine SS model.
Assuntos
Apoptose , Autoantígenos/fisiologia , Glicoproteínas de Membrana/fisiologia , Síndrome de Sjogren/imunologia , Animais , Anticorpos Monoclonais/farmacologia , Autoanticorpos/biossíntese , Autoantígenos/imunologia , Proteínas de Transporte/imunologia , Proteínas de Transporte/fisiologia , Células Cultivadas , Citocinas/biossíntese , Proteína Ligante Fas , Feminino , Imunofenotipagem , Ativação Linfocitária , Metaloproteinase 9 da Matriz/biossíntese , Glicoproteínas de Membrana/imunologia , Camundongos , Proteínas dos Microfilamentos/imunologia , Proteínas dos Microfilamentos/fisiologia , Modelos Biológicos , Síndrome de Sjogren/patologia , Baço/metabolismo , Subpopulações de Linfócitos T/classificação , Linfócitos T/imunologia , Receptor fas/metabolismoRESUMO
OBJECTIVES AND METHODS: Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA-1), which is essential for EBV latency, homodimerizes and binds to the EBV replication origin, oriP. We analyzed the dimerization/DNA-binding domain of EBNA-1 by random and site-directed amino acid substitution. RESULTS: Random point mutations that resulted in reduced DNA binding clustered in the DNA contact region (a.a. 461-473) and at or near the termini of alpha-helix II (514-527). Three substitutions of Gly in the DNA contact region each greatly reduced binding to a single binding site oligonucleotide. Substitutions at and near the termini of alpha-helix II diminished DNA binding. A helix-deforming substitution in alpha-helix I (477-489) blocked DNA binding. A helix-deforming substitution in alpha-helix III (568-582) abolished dimerization and DNA binding. Similarities in surface electrostatic properties and conserved amino acids were found between alpha-helix II and recognition helices of papillomavirus E2 proteins. CONCLUSIONS: The basic DNA contact region is crucial for the specific interaction of EBNA-1 with a single binding site. Alpha-helix I477 is indispensable for oriP binding, and alpha-helix III568 contributes to the homodimeric structure of EBNA-1. Alpha-helix II514 contributes to oriP binding, perhaps changing its alignment with DNA.
Assuntos
Substituição de Aminoácidos/genética , DNA Viral/metabolismo , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Antígenos Nucleares do Vírus Epstein-Barr/química , Antígenos Nucleares do Vírus Epstein-Barr/metabolismo , Herpesvirus Humano 4 , Sequência de Aminoácidos , Sítios de Ligação , Cromatografia Líquida de Alta Pressão , Sequência Conservada/genética , DNA Viral/genética , Proteínas de Ligação a DNA/genética , Dimerização , Antígenos Nucleares do Vírus Epstein-Barr/genética , Glicina/metabolismo , Herpesvirus Humano 4/química , Herpesvirus Humano 4/genética , Modelos Moleculares , Dados de Sequência Molecular , Mutação Puntual/genética , Ligação Proteica , Estrutura Secundária de Proteína , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismo , Origem de Replicação/genética , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Eletricidade Estática , Relação Estrutura-AtividadeRESUMO
OBJECTIVE: Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1) and the replication origin, oriP, are essential for the replication and maintenance of latent EBV DNA in cells, but no enzymatic activity has been associated with EBNA-1 protein alone. In this study, we have searched for host cellular proteins that interact with EBNA-1 protein in various B cell lines latently infected with EBV, including a recently EBV growth-transformed cell line. METHODS: By using gel shift analysis, we investigated the interactions of an oligonucleotide containing a single EBNA-1 recognition site, derived from the family of repeats (FR) element of oriP, with protein from cell extracts. RESULTS: The FR oligonucleotide bound a (72-kD) cellular protein in the absence of EBNA-1 and without induction of the previously reported 'anti-EBNA-1 proteins'. The FR oligonucleotide formed complexes with additional proteins from EBNA-1-synthesizing cell lines; these complexes were abolished or supershifted by anti-EBNA-1 monoclonal antibodies. SDS-PAGE analyses of 35S-Met-labeled proteins that bound to a biotin- conjugated FR oligonucleotide, fractionated by a glycerol gradient centrifugation and affinity-purified with streptavidin, showed three major bands, a 72-kD protein, the FR binding of which seemed to be independent of EBNA-1, a 64-kD protein in both EBNA-1-transfected and latently EBV-infected cell lines, and a 45-kD protein in EBV-infected cell lines, which was most prominent in a recently EBV growth-transformed cell line. CONCLUSIONS: The FR element forms complexes with cellular proteins in the absence and presence of EBNA-1. These 72-, 64- and 45-kD cellular proteins might be involved in the function of the oriP and EBNA-1 system.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Antígenos Nucleares do Vírus Epstein-Barr/fisiologia , Herpesvirus Humano 4/genética , Oligodesoxirribonucleotídeos/metabolismo , Origem de Replicação/genética , Sítios de Ligação , Linfoma de Burkitt , Extratos Celulares , Centrifugação com Gradiente de Concentração , Sequência Consenso/genética , Ensaio de Desvio de Mobilidade Eletroforética , Antígenos Nucleares do Vírus Epstein-Barr/genética , Humanos , Oligodesoxirribonucleotídeos/genética , Ligação Proteica , Transfecção , Células Tumorais CultivadasRESUMO
The ions generated by weak electric current may be used for removal of dental plaque. Also, it has been judged from changes in the viable bacterial cell count and the amount of adenosine triphosphate (ATP) in the saliva that the passage of such a current also has a bactericidal effect on the oral microflora. We confirmed in vitro that 0.5 and 1.0 mA currents that passed for 10 min through phosphate buffered saline containing salivary bacteria were effective in killing the bacteria.
Assuntos
Bactérias , Eletricidade , Saliva/microbiologia , Trifosfato de Adenosina/análise , Soluções Tampão , Contagem de Colônia Microbiana , Humanos , Cloreto de Sódio , Estatísticas não ParamétricasRESUMO
A cleavage product of alpha-fodrin may be an important organ-specific autoantigen in the pathogenesis of Sjogren's syndrome (SS), but the mechanisms of alpha-fodrin cleavage remain unclear. Since EBV has been implicated in the pathogenesis of SS, we determined whether EBV activation could induce the SS-specific 120-kDa autoantigen alpha-fodrin. ZEBRA mRNA expression, a marker for activation of the lytic cycle of EBV, was found in the salivary gland tissues from SS patients, but not in those from control individuals. ZEBRA-expressing lymphoid cells were also found in the SS glands in double-stained immunohistochemistry. Furthermore, a significant link between production of Abs against 120-kDa alpha-fodrin and reactivated EBV Ag was found in sera from patients with SS, but not in those from control individuals. EBV-activated lymphoid cells showed specific alpha-fodrin cleavage to the expected 120-kDa fragments in vitro. Pretreatment with caspase inhibitors inhibited cleavage of alpha-fodrin. Thus, an increase in apoptotic protease activities induced by EBV reactivation may be involved in the progression of alpha-fodrin proteolysis in the development of SS.
Assuntos
Autoantígenos/imunologia , Autoantígenos/metabolismo , Proteínas de Transporte/imunologia , Proteínas de Transporte/metabolismo , Herpesvirus Humano 4/imunologia , Leucina/análogos & derivados , Proteínas dos Microfilamentos/imunologia , Proteínas dos Microfilamentos/metabolismo , Síndrome de Sjogren/imunologia , Clorometilcetonas de Aminoácidos/farmacologia , Apoptose/imunologia , Aprotinina/farmacologia , Autoantígenos/biossíntese , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/biossíntese , Caspases/metabolismo , Inibidores de Cisteína Proteinase/farmacologia , Proteínas de Ligação a DNA/biossíntese , Humanos , Hidrólise , Leucina/farmacologia , Leupeptinas/farmacologia , Proteínas dos Microfilamentos/antagonistas & inibidores , Proteínas dos Microfilamentos/biossíntese , Peso Molecular , Especificidade de Órgãos/imunologia , Pepstatinas/farmacologia , Síndrome de Sjogren/enzimologia , Síndrome de Sjogren/virologia , Transativadores/biossíntese , Células Tumorais Cultivadas/citologia , Células Tumorais Cultivadas/enzimologia , Células Tumorais Cultivadas/metabolismo , Proteínas Virais/biossíntese , Ativação Viral/imunologiaRESUMO
In plastic and reconstructive surgery, postoperative drug therapy with prostaglandin E1 (PGE1) has been widely used to prevent flap necrosis and to extend flap surviving area. In the present study, we assessed the effects of intra-arterial administration of PGE1 on microcirculation. The left cremaster muscle of male Wistar rats were used to measure microcirculatory hemodynamic parameters. The values of internal vessel diameter and erythrocyte velocity were measured by using the confocal laser-scanning microscope (CLSM) system and fluorescent dyes. The blood flow rate was calculated from measured internal vessel diameter and erythrocyte velocity. Internal vessel diameter and flowing erythrocytes were satisfactorily visualized by using the CLSM system. The blood flow rate of arteriole, venule and capillary were dose dependently increased by the intra-arterial administration of PGE1 from the minimum dose of 0.01 ng/kg/min. It is suggested that the intra-arterial administration of PGE1 is a more effective way of postoperative drug therapy than intravenous injection for flap surgery.
Assuntos
Alprostadil/farmacologia , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/cirurgia , Vasodilatadores/farmacologia , Animais , Pressão Sanguínea , Injeções Intra-Arteriais , Masculino , Microcirculação/efeitos dos fármacos , Microscopia/métodos , Ratos , Ratos Wistar , Retalhos CirúrgicosRESUMO
A patient was 31-year-old man with the chief complaint of 38 degrees C fever. He was pointed out left renal tumor by abdominal ultrasonography and computerized tomography (CT). CT revealed left infraclavicular, mediastinal and retroperitoneal lymph nodes swelling and left renal tumor. Serum alpha-fetoprotein (AFP) and beta-human chorionic gonadotropin (HCG-beta) level were elevated. The diagnosis of extragonadal germ cell tumor and left renal cell carcinoma was confirmed pathologically by infraclavicular lymph node and renal biopsy. He was treated with 4 courses of BEP regimen and interferon-alpha, cimetidine therapy for 2 weeks preoperatively. After serum tumor markers were normal level, he underwent left radical nephrectomy and left infraclavicular, mediastinal and retroperitoneal lymph node dissection. The histology of all lymph nodes was necrotic tissue, but operation was incomplete. Therefore VIP therapy was performed postoperatively. This is the first case of extragonadal germ cell tumor coexisted with renal cell carcinoma in the world.
Assuntos
Carcinoma de Células Renais/patologia , Germinoma/patologia , Neoplasias Renais/patologia , Neoplasias do Mediastino/patologia , Neoplasias Primárias Múltiplas , Neoplasias Retroperitoneais/patologia , Adulto , Biomarcadores Tumorais/sangue , Humanos , Linfonodos/patologia , MasculinoRESUMO
Pancreatic cancer frequently causes extrahepatic cholestasis. To identify the direct effects of bile acids in jaundiced serum on pancreatic cancer, the proliferation of PANC-1 and MIA PaCa-2 cells as well as the ultrastructural alteration of PANC-1 cells cultured in crude bile modified media were studied. The growth of these cells in the RPMI-1640 media with or without 1%, 2%, and 4% of the refined crude bile was assessed after 48 and 96 h of incubation. The ultrastructure of PANC-1 cells was investigated by scanning and transmission electron microscopy after 24 and 48 h of incubation. The proliferation of both cell lines in the bile-treated media was greatly inhibited. The inhibitory rates of bile on PANC-1 ranged from 24.1% +/- 3.3% to 66.9% +/- 6.6% (P < 0.01) and those on MIA PaCa-2 ranged from 16.7% +/- 3.8% to 50.7% +/- 5.5%. (P < 0.01). When the bile-added media were replaced, the cells were able to restore their proliferating ability. The PANC-1 cells incubated in the bile-supplied media indicated that the mirovilli, mitochondria, and other organelles had thus been injured. These results suggest that bile acids appear to inhibit the proliferation of PANC-1 and MIA PaCa-2 cells, and the probable inhibitory mechanism is mainly considered to be due to the cytotoxicity of such bile acids.
Assuntos
Ácidos e Sais Biliares/metabolismo , Colestase Extra-Hepática/metabolismo , Citotoxinas/metabolismo , Neoplasias Pancreáticas/metabolismo , Divisão Celular , Colestase Extra-Hepática/etiologia , Colestase Extra-Hepática/patologia , Meios de Cultivo Condicionados , Inibidores do Crescimento/metabolismo , Humanos , Técnicas In Vitro , Neoplasias Pancreáticas/complicações , Neoplasias Pancreáticas/patologia , Células Tumorais Cultivadas/metabolismoRESUMO
Here, we report a case of a two-day-old neonate with total anomalous pulmonary venous connection to the innominate vein and a bronchogenic cyst arising from the trachea. Antenatal echocardiography had delineated both cardiac and extracardiac lesions, and a repeated examination on the day of birth disclosed progressive enlargement in the cyst in a manner so as to obstruct the innominate vein. On the second day of life, the patient underwent complete correction of the cardiac lesion and total excision of the cyst. The patient recovered uneventfully and was discharged on the thirteenth postoperative day.
Assuntos
Cisto Broncogênico/complicações , Veias Pulmonares/anormalidades , Veias Braquiocefálicas/anormalidades , Cisto Broncogênico/cirurgia , Procedimentos Cirúrgicos Cardíacos/métodos , Humanos , Recém-Nascido , Masculino , Veias Pulmonares/cirurgiaRESUMO
The liver is the most common site of metastasis in pancreatic cancer, and there are no promising strategies to treat it. Angiostatin, a kringle-containing fragment of plasminogen, is a potent inhibitor of angiogenesis. The effect of angiostatin on liver metastasis in pancreatic cancer was investigated by using our established hamster model of liver metastasis. Pancreatic cancer cells (PGHAM-1, 1 x 10(6)) derived from N-nitrosobis(2-oxopropyl)amine (BOP)-induced pancreatic tumor in Syrian golden hamsters were transplanted into the spleen of female hamsters, and the animals were subcutaneously injected with angiostatin and saline. Subsequently, the macroscopic appearance of liver surface metastases was evaluated. In addition, histological sections of the liver metastases were analyzed for neovascularization, proliferation, and apoptosis on the basis of von Willebrand factor, argyrophilic nucleolar organizer region (Ag-NOR), and TdT-mediated dUTP-biotin nick end labeling (TUNEL) staining, respectively. The results showed significant tumor growth retardation and inhibition of angiogenesis in metastatic liver tumors in response to treatment with angiostatin. Moreover, the metastases remained in a nearly dormant state due to a balance between apoptosis and proliferation of the tumor, with no detectable side effects. This is the first experimental trial of angiostatin on pancreatic cancer and liver metastasis. The results suggest that angiostatin therapy could be effective against liver metastases of pancreatic cancer.
Assuntos
Inibidores da Angiogênese/farmacologia , Antineoplásicos/farmacologia , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Neoplasias Hepáticas Experimentais/secundário , Neovascularização Patológica/tratamento farmacológico , Neoplasias Pancreáticas/patologia , Fragmentos de Peptídeos/farmacologia , Plasminogênio/farmacologia , Inibidores da Angiogênese/toxicidade , Angiostatinas , Animais , Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Cricetinae , Feminino , Imuno-Histoquímica , Neoplasias Hepáticas Experimentais/irrigação sanguínea , Mesocricetus , Transplante de Neoplasias , Neoplasias Pancreáticas/tratamento farmacológico , Fragmentos de Peptídeos/toxicidade , Plasminogênio/toxicidade , Células Tumorais CultivadasRESUMO
We created anexperimental model of pulmonary metastasis based on subcutaneously implanted Lewis lung cancer in mice and observed in vivo the microcirculation of spontaneously metastasized tumors in the lung. The mice lung was held by a small handmade suction ring to stop cardiac and respiratory movement. Using fluorescent microscopy, tumor microcirculation and normal lung microcirculation in the same lung lobe were compared by measuring microvessel diameter and blood flow velocity [red blood cell (rbc) velocity]. In normal microcirculation, the mean values of microvessel diameter and rbc velocity were 10.4 +/- 2.7 microm and 188 +/- 63 microm/s, respectively. In tumor microcirculation, the mean values of the same were 10.6 +/- 3.3 microm and 105 +/- 40 microm/s. The rbc velocity in normal microcirculation was significantly higher (P < 0.001) than that in tumor microcirculation. The calculated shear rates of normal microcirculation and tumor microcirculation were 73.4 +/- 23.4 (/s) and 41.2 +/- 16.1 (/s), respectively. The shear rate of the tumor microcirculation was significantly slower (P < 0.001) than that of the normal microcirculation. We demonstrated a feasibility of observation and measurement of tumor microcirculation in the lung and confirmed that the physiologic data were compatible to those in the brain or in the liver reported by others. This model might be useful for studying metastatic tumor pathophysiology in the lung microcirculation.
Assuntos
Carcinoma Pulmonar de Lewis/secundário , Neoplasias Pulmonares/secundário , Circulação Pulmonar , Animais , Carcinoma Pulmonar de Lewis/irrigação sanguínea , Dextranos/farmacocinética , Estudos de Viabilidade , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/farmacocinética , Corantes Fluorescentes/farmacocinética , Hemorreologia , Injeções Subcutâneas , Neoplasias Pulmonares/irrigação sanguínea , Camundongos , Camundongos Endogâmicos C57BL , Microcirculação , Transplante de NeoplasiasRESUMO
A 2-year old boy visited our clinic with a chief complaint of high fever. A past history of acute renal failure due to cystine stones and cystinuria was expressed. Abdominal rentogenograms and CT demonstrated a right ureteral stone and a left renal stone. Furthermore renogram evaluation indicated non-function of the right kidney and dysfunction in the left kidney. Since right ureteral stone moved into bladder seven days post-admission, right ureteroscopy, left PNL, and cystolithotripsy were performed. Considering that right ureteral stenosis was determined by ureteroscopy, balloon dilation against the stenotic ureteral wall was performed. Left PNL and cystolithotripsy were successfully performed. No intraoperative complications occurred and no symptoms of signs of recurrence of the underlying metabolic disease were evident four months postoperatively.
Assuntos
Cistina/análise , Cálculos Renais/terapia , Rim/fisiopatologia , Litotripsia/métodos , Pré-Escolar , Humanos , Cálculos Renais/química , Cálculos Renais/fisiopatologia , Masculino , Resultado do TratamentoRESUMO
When we evaluated the age-associated changes in autoimmune exocrinopathy in a NFS/sld murine model for primary Sjögren's syndrome (SS), severe destructive autoimmune lesions developed in the salivary and lacrimal glands in the aged mice, compared with those observed in the younger model. We detected a decreased secretion of saliva and tear flow in the aged group. A significant increase of TUNEL(+)-apoptotic epithelial duct cells in the salivary glands was detected in the aged SS animal model. A higher proportion of mouse salivary gland cells bearing Fas was found in the aged group, whereas no significant changes were seen on tissue-infiltrating CD4(+) T cells bearing FasL in the salivary glands from young and aged mice. We detected an increased cleavage product of organ-specific autoantigen, 120-kd alpha-fodrin, in the aged salivary gland tissues on immunoblotting, and an increase in serum autoantibody production against 120-kd alpha-fodrin by enzyme-linked immunosorbent assay. An increase in the proliferative response of splenic T cells against organ-specific autoantigen was observed, whereas nonspecific concanavalin A responsiveness was decreased in the aged mice. In addition, a decrease in Fas expression was found on splenic CD4(+) T cells in the aged mice, and anti-Fas mAb-stimulated apoptosis was down-regulated on CD4(+) T cells. These results indicate that age-associated dysregulation of CD4(+) T cells may play a crucial role on acceleration of organ-specific autoimmune lesions in a murine model for primary SS through Fas-mediated apoptosis.
Assuntos
Envelhecimento , Apoptose , Doenças Autoimunes/patologia , Síndrome de Sjogren/patologia , Receptor fas/fisiologia , Animais , Autoanticorpos/sangue , Autoantígenos/metabolismo , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Feminino , Marcação In Situ das Extremidades Cortadas , Masculino , Camundongos , Saliva/metabolismo , Lágrimas/metabolismo , Receptor fas/imunologiaRESUMO
OBJECTIVES: To determine the usefulness of the saccharin time (ST) test for evaluating the mucociliary function of the maxillary sinus after endoscopic sinus surgery (ESS) for chronic sinusitis. METHODS: This study was conducted on 88 maxillary sinuses of 74 patients after ESS. The maxillary sinus fontanel was broadly opened via the middle meatus using an endoscope, and a saccharin granule was adhered to the bottom of the maxillary sinus mucosa The time until the patient recognized the sweet taste was recorded. Before the ST test, the bilateral maxillary sinuses were classified into the following four groups on the basis of the post-ESS severity of mucosal edema and swelling as revealed by endoscopic observation: normal (45 sinuses), mild mucosal edema and swelling (24), moderate mucosal finding (14), and severe mucosal finding or filling of the sinus with a polyp(s) (5). RESULTS: The mean ST values in the normal group and the groups with mild, moderate, and severe mucosal edema and swelling were 35.7, 38.1, 63.6, and 88.0 minutes, respectively. Thus the ST increased with the post-ESS severity of the mucosal lesion. However, for the group with mild mucosal edema and swelling, scanning electron microscopic observation of three maxillary sinuses in which the ST exceeded 120 minutes and four sinuses in which the ST was 40 minutes revealed extensive cilia loss in the former sinuses, but not in the latter. A second post-ESS endoscopic observation was performed in 17 patients, revealing improvement in 11 sinuses, no change in 5 sinuses, and aggravation in 1 sinus (compared with the initial test). The ST test was also repeated, revealing that the ST became shorter in most of the endoscopically improved sinus group. However, a few sinuses showed a discrepancy between the change in the endoscopic findings and the ciliary function (ST). CONCLUSION: Measurement of the maxillary sinus ST is a simple, accurate, and useful technique for assessing the post-ESS mucociliary function in conjunction with endoscopy, and the information gained can help in deciding subsequent therapy.