RESUMO
BACKGROUND: Non-Helicobacter pylori helicobacters (NHPHs) besides H. pylori infect human stomachs and cause chronic gastritis and mucosa-associated lymphoid tissue lymphoma. Cholesteryl-α-glucosides have been identified as unique glycolipids present in H. pylori and some Helicobacter species. Cholesterol-α-glucosyltransferase (αCgT), a key enzyme for the biosynthesis of cholesteryl-α-glucosides, plays crucial roles in the pathogenicity of H. pylori. Therefore, it is important to examine αCgTs of NHPHs. MATERIALS AND METHODS: Six gastric NHPHs were isolated from Japanese patients and maintained in mouse stomachs. The αCgT genes were amplified by PCR and inverse PCR. We retrieved the αCgT genes of other Helicobacter species by BLAST searches in GenBank. RESULTS: αCgT genes were present in most Helicobacter species and in all Japanese isolates examined. However, we could find no candidate gene for αCgT in the whole genome of Helicobacter cinaedi and several enterohepatic species. Phylogenic analysis demonstrated that the αCgT genes of all Japanese isolates show high similarities to that of a zoonotic group of gastric NHPHs including Helicobacter suis, Helicobacter heilmannii, and Helicobacter ailurogastricus. Of 6 Japanese isolates, the αCgT genes of 4 isolates were identical to that of H. suis, and that of another 2 isolates were similar to that of H. heilmannii and H. ailurogastricus. CONCLUSIONS: All gastric NHPHs examined showed presence of αCgT genes, indicating that αCgT may be beneficial for these helicobacters to infect human and possibly animal stomachs. Our study indicated that NHPHs could be classified into 2 groups, NHPHs with αCgT genes and NHPHs without αCgT genes.
Assuntos
Glucosiltransferases/genética , Infecções por Helicobacter/microbiologia , Helicobacter/enzimologia , Helicobacter/genética , Linfoma de Zona Marginal Tipo Células B/microbiologia , Animais , Feminino , Gastrite/microbiologia , Gastrite/patologia , Genoma Bacteriano/genética , Helicobacter/classificação , Infecções por Helicobacter/patologia , Humanos , Japão , Linfoma de Zona Marginal Tipo Células B/patologia , Camundongos , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Virulência/genéticaRESUMO
Soybean cyst nematode (SCN) Heterodera glycines Ichinohe, a plant parasite, is one of the most serious pests of soybean. In this paper, we report that SCN is attracted to nitrate and its analogs. We performed attraction assays to screen for novel attractants for SCN and found that nitrates were attractants for SCN and SCN recognized nitrate gradients. However, attraction of SCN to nitrates was not observed on agar containing nitrate. To further elucidate the attraction mechanism in SCN, we performed attraction assays using nitrate analogs ([Formula: see text], [Formula: see text], [Formula: see text]). SCN was attracted to all nitrate analogs; however, attraction of SCN to nitrate analogs was not observed on agar containing nitrate. In contrast, SCN was attracted to azuki root, irrespective of presence or absence of nitrate in agar media. Our results suggest that the attraction mechanisms differ between plant-derived attractant and nitrate.
Assuntos
Fatores Quimiotáticos/farmacologia , Nitratos/farmacologia , Tylenchoidea/efeitos dos fármacos , Ágar/farmacologia , Animais , Fatores Quimiotáticos/química , Nitratos/química , Doenças das Plantas/parasitologia , Doenças das Plantas/prevenção & controle , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/parasitologia , Glycine max/efeitos dos fármacos , Glycine max/parasitologia , Relação Estrutura-Atividade , Tylenchoidea/fisiologiaRESUMO
Soybean cyst nematode (SCN), Heterodera glycines Ichinohe, is a plant-parasitic nematode and one of the most serious soybean pests. Herein, we present the heterocyclic compound 1,10-phenanthroline (Phen) and its derivatives as novel hatching stimulants for SCN. Phen treatment promoted hatching of second-stage juveniles of SCNs in a concentration-dependent manner. In addition, the hatching of SCNs following treatment with Phen occurred more rapidly than that following treatment with the known hatching stimulant, glycinoeclepin A (GEA). Furthermore, the co-application of Phen and GEA enhanced SCN hatching rate compared with that of Phen or GEA alone. A structure-activity relationship study for Phen derivatives suggested that 2,2'-bipyridine is the essential structure of the SCN-hatching stimulants. These results suggest that Phen and its derivatives activate different hatching pathways of SCNs from GEA.
Assuntos
Glycine max/parasitologia , Nematoides/crescimento & desenvolvimento , Fenantrolinas/farmacologia , Animais , Feminino , Relação Estrutura-AtividadeRESUMO
Cardiac amyloidosis is a manifestation of one of several systemic amyloidoses, and is characterized by increased left-ventricular (LV) wall thickness and normal or decreased LV cavity size. Congestive heart failure in cardiac amyloidosis is characterized by a predominant diastolic LV dysfunction, and systolic dysfunction occurs only in late-stage disease. Echocardiography is a noninvasive, reproducible method for assessing cardiac morphology and function in cardiac amyloidosis, and some echocardiographic indices are prognostic for amyloidoses. This review describes the advances in echocardiography and its role in the diagnosis and management of cardiac amyloidoses. Our review suggests that LV longitudinal function and the cyclic variation of myocardial integrated backscatter may be the best predictors of adverse outcomes. In the future, new echocardiographic techniques, such as fully automated echocardiogram interpretation, should provide further useful information for assessing cardiac function and prognosis in cardiac amyloidosis patients.
Assuntos
Amiloidose , Cardiomiopatias , Ecocardiografia , Ventrículos do Coração/diagnóstico por imagem , Amiloidose/diagnóstico , Amiloidose/fisiopatologia , Cardiomiopatias/diagnóstico , Cardiomiopatias/fisiopatologia , Ecocardiografia/métodos , Ecocardiografia/tendências , Ventrículos do Coração/patologia , Ventrículos do Coração/fisiopatologia , Humanos , PrognósticoRESUMO
Cutaneous Epstein-Barr virus (EBV)-associated B-cell lymphoma (EBVBL) in non-immunocompromised patients is very rare. Here, we report a case of cutaneous EBVBL in a 72-year-old Japanese woman without any signs of immunosuppression. She showed repeated high fever and skin eruptions on the face, limbs and palms. Histological diagnosis was diffuse large B-cell lymphoma. EBV infection was detected by in situ hybridization and Southern blotting. Immunostaining for viral proteins showed the patient to be positive for latent membrane protein 1 (LMP-1) and negative for Epstein-Barr virus nuclear antigen-1 (EBNA-2), indicating that a type II latency EBV infection pattern.
Assuntos
Infecções por Vírus Epstein-Barr/imunologia , Herpesvirus Humano 4/isolamento & purificação , Linfoma Difuso de Grandes Células B/virologia , Neoplasias Cutâneas/virologia , Idoso , Idoso de 80 Anos ou mais , Antígenos CD/análise , DNA Viral/análise , Infecções por Vírus Epstein-Barr/complicações , Evolução Fatal , Feminino , Herpesvirus Humano 4/genética , Humanos , Imunocompetência , Técnicas Imunológicas , Linfoma Difuso de Grandes Células B/imunologia , Linfoma Difuso de Grandes Células B/patologia , Masculino , Pessoa de Meia-Idade , RNA Viral/análise , Neoplasias Cutâneas/imunologia , Neoplasias Cutâneas/patologia , Proteínas da Matriz Viral/análise , Latência ViralRESUMO
We evaluated three test kits for fibrin degradation products (FDP) D-dimer. We found that six of 217 plasma sample values obtained by Nanopia test were markedly higher than the values obtained using the other two kits. The regression equation for 211 samples (excluding six) was y=0.64x+3.05 (y: Nanopia, x: LIAS AUTO) and the correlation coefficient was 0.915. Therefore, we classified these samples into three categories, namely correlated(y< 1.0x), incompatible (y= 1.0x-2.9x) and markedly incompatible (y> or =3.0x). Selected samples, eight correlated, four incompatible and four markedly incompatible, were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western blotting(WB). WB analysis using anti-fibrinogen antibody showed that both high molecular weight fragments of cross-linked fibrin (HMW-XDP) and DD/E fragments were present in the correlated samples, but there was less HMW-XDP than DD/E in the incompatible samples and mostly DD/E (HMW-XDP was significantly less than DD/E) in the markedly incompatible samples. These data suggest that plasma FDP samples that contain mostly DD/E and little HMW-XDP demonstrated markedly incompatible values using the three D-dimer test kits. These data was reflected by markedly elevated plasmin alpha2-plasmin inhibitor complex values in the incompatible and markedly incompatible samples. Unfortunately, we did not directly demonstrate these phenomena by WB analysis with two anti-D-dimer antibodies used Nanopia or LPIA reagent. In the near future, we expect that standardization of FDP D-dimer assay will be accomplished.
Assuntos
Produtos de Degradação da Fibrina e do Fibrinogênio/imunologia , Anticorpos Monoclonais/imunologia , Western Blotting , Produtos de Degradação da Fibrina e do Fibrinogênio/análise , Humanos , Kit de Reagentes para DiagnósticoRESUMO
Chimerism analysis by polymerase chain reaction amplification of short tandem repeats (PCR-STR) has become a routine diagnostic procedure for evaluating grafts and assessing the likeliness of original disease recurrence after allogeneic stem cell transplantation. Following a sex-mismatched hematopoietic stem cell transplantation (HSCT), we monitored the clinical course of a 61-year old male AML M6 patient with trisomy 8 using PCR-STR with a TH01 locus on 11p15 and fluorescence in situ hybridization (FISH) analysis specific for alpha satellite DNA on chromosome 8. Ten months after HSCT, FISH analysis showed 24.8% recipient cells, but PCR-STR demonstrated 100% donor type chimerism. Further XY FISH analysis of May-Grünwald-Giemsa-stained bone marrow samples clearly demonstrated relapse of the original disease and G-banding analysis of bone marrow samples at relapse showed that an additional chromosomal abnormality, del(11) (p10), had deleted the PCR-STR detection site in all recipient type cells. As such, clinicians should consider the possibility that unexpected karyotype changes may invalidate PCR-STR analysis findings, especially when conflicting results appear among chimerism analyses.
Assuntos
Quimerismo , Cromossomos Humanos Par 8/genética , Transplante de Células-Tronco Hematopoéticas , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/terapia , Humanos , Hibridização in Situ Fluorescente , Masculino , Repetições de Microssatélites/genética , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Recidiva , Transplante HomólogoRESUMO
Esophageal gland duct adenomas are extremely rare tumors. Here, we report the case of a 75-year-old Japanese man who had undergone total gastrectomy for advanced gastric cancer. Esophageal gland duct adenoma was incidentally found in the lower esophagus. It appeared to be detached from the site of gastric cancer and was well demarcated without a capsule. Histologic analysis revealed papillary and cystic structures mainly comprising eosinophilic cells with minimum nuclear atypia. Immunohistochemical analysis revealed that the tumor were diffusely positive for the S100 protein with preserved alpha-SMA-positive myoepithelial cell layers and a characteristic cytokeratin expression pattern similar to that in normal esophageal gland ducts (CK5/6+++, CK7+++, CK17+, CK18+, CK19+++, CK20-, HMWCK+++). In addition, differentiation into the terminal duct was confirmed by a combination of mucin staining and immunohistochemical and ultrastructural examinations. This is the first report that refers to the ultrastructural findings of an esophageal gland duct adenoma and describes terminal duct differentiation. We believe that the possibility of an esophageal gland duct adenoma should be considered when diagnosing a ductal or glandular lesion of the esophagus.
Assuntos
Adenoma/patologia , Neoplasias Esofágicas/patologia , Esôfago/patologia , Adenocarcinoma/patologia , Adenocarcinoma/cirurgia , Adenoma/química , Idoso , Biomarcadores Tumorais/análise , Cárdia/patologia , Cárdia/cirurgia , Núcleo Celular/ultraestrutura , Citoplasma/ultraestrutura , Neoplasias Esofágicas/química , Esôfago/química , Humanos , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Transmissão , Mucosa/química , Mucosa/patologia , Segunda Neoplasia Primária , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgiaRESUMO
BACKGROUND: Pathological alteration in gastric mucosa is caused by Helicobacter pylori infection and is detectable by histological analysis. In particular, the alteration of gland mucous cells (GMCs)-type mucin, which plays a protective role against H. pylori infection, is critical in the pathogenesis of H. pylori-related gastritis. We established an assay for GMCs-type mucin and quantitatively assessed the pathophysiological changes in its content in human gastric juice samples. METHODS: The assay method for GMCs-type mucin was based on ELISA using a monoclonal antibody (HIK1083), and was used it to measure GMCs-type mucin in gastric juice obtained from patients with or without H. pylori infection. RESULTS: All the basic characteristics of the current method were satisfactory to quantify the GMCs-type mucin content in gastric juice. The GMCs-type mucin content, but not total mucin content, was significantly higher in patients with H. pylori infection (n=17; 437+/-476 U, mean+/-SD) than in those without H. pylori infection (n=55; 168+/-322 U, p<0.05). CONCLUSIONS: The current method is suitable for the quantitative analysis of GMCs-type mucin in gastric juice. The change in GMCs-type mucin content in gastric juice may be possibly implicated in the pathophysiology of the gastric mucosa and in the patient's gastric mucosal lesions.
Assuntos
Anticorpos Monoclonais/imunologia , Suco Gástrico/metabolismo , Mucinas Gástricas/análise , Mucosa Gástrica/metabolismo , Mucosa Gástrica/fisiopatologia , Gastropatias/metabolismo , Gastropatias/fisiopatologia , Animais , Calibragem , Ensaio de Imunoadsorção Enzimática , Suco Gástrico/imunologia , Mucinas Gástricas/imunologia , Mucinas Gástricas/metabolismo , Mucosa Gástrica/imunologia , Humanos , Concentração de Íons de Hidrogênio , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Gastropatias/imunologia , Suínos , TemperaturaRESUMO
We previously reported a case of heterozygous beta-thalassaemia with IVS1-1G > C substitution in the beta-globin gene and a non-detectable level of mutant mRNA in the patient's reticulocytes. The purpose of this study was to determine whether the transcription and RNA splicing and processing of the mutant gene occurred. We analysed the expression of the mRNA encoded by the cloned mutant gene in COS-1 cells by reverse transcription-polymerase chain reaction followed by agarose gel electrophoresis and nucleotide sequencing. The G > C mutation completely inactivated the normal 5' splice site and resulted in the activation of two cryptic 5' splice sites, located 16 and 38 nt upstream of the normal site. The usage of these two cryptic sites accords with the findings of reports on IVS1-1G > A or IVS1-1G > C substitution of exon 1 of the beta-globin gene. Additional experiments that involved transfection of equal amounts of both normal and mutant vectors into COS-1 cells indicated the presence of mutant mRNAs. In conclusion, the beta-thalassaemia gene (IVS1-1G > C) was expressed in transfected cells, but showed aberrant RNA splicing. Further studies will be required to clarify the molecular mechanism that results in severe reduction in the mutant mRNA level in vivo.
Assuntos
Processamento Alternativo/genética , Globinas/genética , Polimorfismo de Nucleotídeo Único , Sítios de Splice de RNA/genética , Talassemia beta/genética , Animais , Sequência de Bases , Células COS , Chlorocebus aethiops , Clonagem Molecular , Expressão Gênica/fisiologia , Humanos , RNA Mensageiro/metabolismo , TransfecçãoRESUMO
This study used matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) to identify all lipid classes in human serum lipoproteins. After the major lipoproteins classes were isolated from serum by ultracentrifugation, the lipids were extracted and mixed with 2,5-dihydroxybenzoic acid (2,5-DHB) dissolved in Folch's solution (chloroform/methanol 2:1, v/v). MALDI-TOF MS analysis of the samples identified phospholipids (PLs), lysophospholipids (lysoPLs), sphingolipids (SLs), triglycerides (TGs), cholesteryl esters (CEs), and free cholesterol; it also showed the characteristics of individual fatty acid chains in serum lipids. MALDI-TOF MS allowed analysis of strongly hydrophobic and non-polar molecules such as CEs and TGs as well as hydrophilic molecules such as phospholipids. Direct analysis of fatty acids was not possible. The concentrations of lipids were not consistent with the ion peak intensities, since the extent of polarity affected the ionization characteristics of the molecules. However, lipid molecules with similar molecular structures but various fatty acid chains, such as phosphatidylcholine (PCs), were analyzed quantitatively by MALDI-TOF MS. Quantitative measurement of cholesterol was possible with the use of an internal standard. This study shows that MALDI-TOF MS can be used for direct investigation and quantitative analysis of the phospholipid composition of serum lipoproteins.
Assuntos
Lipoproteínas/análise , Lipoproteínas/sangue , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Adulto , Colesterol/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fosfolipídeos/análiseRESUMO
We analyzed the clinical factors resulting in hypofibrinogenemia, which is defined as less than 100mg/dl of plasma fibrinogen values determined by a procedure based on the Thrombin-time method. Within a 12-month period, we assayed 5,746 patients (19,309 plasmas) and found 113 patients (1.97%) with hypofibrinogenemia. We categorized these patients as having decreased synthesis of fibrinogen (less than 3.0g/dl of albumin, 140 IU/l of Cholinesterase, and/or 50% on Hepaplastin Test), increased consumption of fibrinogen (more than 10 microg/ml of FDP D-dimer), known side effect of L-asparaginase administration, or other causes. Details are follows: 1) decreased synthesis: 26 patients, suspected of decreased synthesis (albumin: 3.1-3.4 g/dl): 4 patients, 2) increased consumption: 15 patients, suspected of increased consumption (FDP D-dimer: 5.0-9.9 g/dl): 1 case, 3) decreased synthesis combined with increased consumption: 24 patients, suspected of decreased synthesis and/or suspected of increased consumption: 14 patients, 4) side-effect of L-asparaginase administration: 24 patients, 5) heterozygous dysfibrinogenemia: 1 patient, 6) heterozygous fibrinogen deficiency: 1 patient, suspected of heterozygous fibrinogen deficiency: 1 patient, 7) unidentified: 2 patients with West syndrome treated with a combination of ACTH and valproic acid. Three patients with dysfibrinogenemia or fibrinogen deficiency showed normal or slightly prolonged PT values and normal APTT values. These data and our previous reports suggest that heterozygous patients with dysfibrinogenemia or fibrinogen deficiency do not demonstrate markedly prolonged PT and APTT values, differing from patients with afibrinogenemia.
Assuntos
Afibrinogenemia/diagnóstico , Afibrinogenemia/etiologia , Tempo de Trombina , Hormônio Adrenocorticotrópico/efeitos adversos , Afibrinogenemia/genética , Asparaginase/efeitos adversos , Fibrinogênio/biossíntese , Fibrinogênio/metabolismo , Heterozigoto , Humanos , Lactente , Espasmos Infantis/complicações , Ácido Valproico/efeitos adversosRESUMO
Structural abnormalities involving the mixed-lineage leukemia (MLL) gene on 11q23 have been associated with hematological malignancies. The rearrangement of MLL occurs during translocations and insertions involving a variety of genes on the partner chromosome. We report a rare case of acute myelogenous leukemia (AML-M2) with 11q23 abnormalities. Fluorescence in situ hybridization (FISH) using a commercial dual-color MLL probe detected an atypical signal pattern: one fusion signal, two green signals smaller than those usually detected, and no orange signals. Spectral karyotyping (SKY) analysis indicated that one green signal was detected on the short arm of derivative chromosome 10, and the other green signal on the long arm of a derivative chromosome 11, on which no orange signal was detected. A long-distance inverse polymerase chain reaction (LDI-PCR) identified the fusion partner gene, in which intron 6 of MLL was fused with intron 8 of AF10 on 10p12 in the 5' to 3' direction. Our observations indicated that the MLL-AF10 fusion gene resulted from the insertion of part of the region that included the 5' MLL insertion into 10p12; this was concurrent with the deletion of 3' MLL.
Assuntos
Cromossomos Humanos Par 10/genética , Deleção de Genes , Leucemia Mieloide/patologia , Proteína de Leucina Linfoide-Mieloide/genética , Proteínas de Fusão Oncogênica/genética , Doença Aguda , Adulto , Sequência de Aminoácidos , Sequência de Bases , Aberrações Cromossômicas , Bandeamento Cromossômico , Quebra Cromossômica , Deleção Cromossômica , Cromossomos Humanos Par 11/genética , Histona-Lisina N-Metiltransferase , Humanos , Hibridização in Situ Fluorescente/métodos , Cariotipagem , Leucemia Mieloide/genética , Masculino , Mutagênese Insercional/genética , Proteína de Leucina Linfoide-Mieloide/química , Análise de Sequência de DNA , Cariotipagem Espectral/métodos , Fatores de Transcrição/genéticaRESUMO
Clinical laboratories need to know and manage the costs of laboratory tests, because they need financial data (1) to estimate costs per patient, (2) to request a budget to buy equipment, and (3) to improve their work; however, less than 40% laboratories practice cost management. In 2002, Shinshu University Hospital began to assess the costs of laboratory tests, but it was difficult to evaluate the quality of our cost management because there are few data and papers about the costs of laboratory tests in Japan. In this article, we practiced cost analysis using Shinshu University Hospital's data for 3 years (2002-2004), and studied the features of laboratory test costs and the problems of laboratory cost management. As a result, we listed 7 points to check cost management in clinical laboratories. This check list was established using only one data from our hospital. So, we suggest the benchmarking laboratory test costs between laboratories of the same type of hospitals or various laboratories.
Assuntos
Técnicas de Laboratório Clínico/economia , Custos e Análise de Custo , Laboratórios Hospitalares/economia , Benchmarking/economia , Hospitais Universitários/economia , Humanos , Japão , Serviços Terceirizados/economia , Sistema de Pagamento Prospectivo/economiaRESUMO
Polysomnography (PSG) is the gold standard for the diagnosis of sleep apnea syndrome (SAS). However, PSG is not suitable as a first-line examination for all people suspected as having SAS because PSG requires hospitalization. Therefore, it is hoped that a simple examination can be developed which is available for use in the home. The present study evaluated the usefulness of a new sheet-like apparatus (SD-101) which is equipped with 162 pressure sensors for SAS diagnosis. One hundred patients hospitalized for PSG were simultaneously examined using the SD-101, and 25 patients, who underwent both PSG and SAS screening with MORPHEUS R, were also studied. The SD-101 is inserted between a sheet and the bed, and detects pressure from many points on the patient's body as it presses against the bed. Continuous changes of these pressure points are converted to respiratory movement. A very close correlation was seen between the apnea hypopnea index of PSG and a respiratory disturbance index of SD-101 (r=0.90), although there was a significant but lower correlation between data obtained PSG and MORPHEUS R(r=0.84). The sensitivity and specificity of the examination using SD-101 were 98.2% and 55.8%, respectively. These findings suggested that a new apparatus, SD-101, may be useful for the screening of SAS.
Assuntos
Manometria/instrumentação , Síndromes da Apneia do Sono/diagnóstico , Roupas de Cama, Mesa e Banho , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , PolissonografiaRESUMO
The hypodysfibrinogenemia Otsu is caused by the two-residue deletion, gammaAsn319 and gammaAsp320. Analysis of plasma or purified fibrinogen from the heterozygous propositus revealed that the amount of variant gamma-chain was lower than that of normal gamma-chain. In order to examine the basis for this difference, we transfected Chinese hamster ovary cells and established stable cell lines that expressed both chains, gammaDelta/gammaN, only the normal chain, gammaN, and only the variant chain, gammaDelta. We measured fibrinogen concentration of confluent cultures by ELISA. We found the ratios of the concentrations in the media to the concentrations in the cell lysates of gammaDelta, gammaDelta/gammaN, and gammaN-cells were 0.42, 0.60, and 1.00, respectively. We measured the concentrations of the gammaDelta and gammaN chains by densitometric analysis of samples following separation by SDS-PAGE and found the fraction of gammaDelta-chains in cell lysates was always greater than the fraction in the respective culture media. We examined the kinetics of fibrinogen synthesis, assembly and secretion in pulse-chase experiments, and found that the gammaDelta-chain was assembled into intact fibrinogen at a rate similar to assembly of the gammaN-chain into normal fibrinogen, but was secreted into the medium at a slightly slower rate than normal fibrinogen. Considered together, these experiments indicate secretion of the variant fibrinogen was slightly impaired. These results suggest that the reduced level of gammaDelta319,320 fibrinogen in the plasma of the Otsu patient arises from modestly impaired secretion of this variant fibrinogen.
Assuntos
Fibrinogênio/genética , Fibrinogênios Anormais/metabolismo , Deleção de Genes , Animais , Western Blotting , Células CHO , Cricetinae , Densitometria , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Fibrinogênio/química , Fibrinogênios Anormais/genética , Vetores Genéticos , Heterozigoto , Humanos , Imunoensaio , Immunoblotting , Técnicas In Vitro , Cinética , Metionina/química , Proteínas Recombinantes/química , Fatores de Tempo , TransfecçãoRESUMO
Helicobacter pylori (Hp) infection is an important factor in human gastric disorders, including chronic active gastritis, peptic ulcers, intestinal metaplasia and cancer. Since epidemiologic studies overwhelmingly agree on a protective influence of fruits and vegetables in reducing the risk of gastric neoplasia and processed foods made from Prunus mume Sieb. et Zucc. (Japanese apricot or "Ume" in Japanese) are traditionally known for their miscellaneous medical effects, in the present study we investigated the efficacy of a fruit-juice concentrate of Japanese apricot (CJA) in the glandular stomach of Hp-infected Mongolian gerbils. Hp-inoculated gerbils were given CJA in their drinking water at concentrations of 1 and 3% for 10 weeks. The microscopic scores for gastritis and mucosal hyperplasia in the CJA groups were significantly lower than in the Hp-inoculated control group, with dose-dependence. Real-time PCR was performed to quantitate Hp by demonstrating urease A gene amount using gerbils glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene as an internal control. Average relative urease A gene dosage in the glandular stomach in the 1 and 3% CJA and Hp-inoculated control groups was 26.6 +/- 11.6% (average +/- SE), 30.3 +/- 10.5%, 100 +/- 40.9%, respectively, the fruit-juice concentrate causing significant lowering (P<0.01 and P<0.05, respectively, with 1 and 3%). These findings suggest that suppressive effects on gastric cancer development might also be expected as a result of decreased numbers of Hp and improvement of Hp-induced chronic active gastritis on administration of CJA.
Assuntos
Bebidas , Gastrite/microbiologia , Gastrite/prevenção & controle , Infecções por Helicobacter/complicações , Extratos Vegetais/farmacologia , Prunus/química , Neoplasias Gástricas/microbiologia , Neoplasias Gástricas/prevenção & controle , Administração Oral , Animais , Gerbillinae , Gliceraldeído 3-Fosfato Desidrogenase (NADP+)/genética , Gliceraldeído 3-Fosfato Desidrogenase (NADP+)/metabolismo , Helicobacter pylori/patogenicidade , Inflamação , Masculino , Reação em Cadeia da PolimeraseRESUMO
We compared the seroepidemiologic patterns of Helicobacter pylori and hepatitis A virus (HAV) infections among participants in 2 independent cross-sectional studies conducted in Japan in 1986 and 1994. Subgroups were monitored with successive blood sampling. H. pylori and HAV infection status was defined by results of enzyme-linked immunosorbent assay. In 1986, the prevalence of H. pylori infection and HAV infection, respectively, were 80% and 70% among adults and 31% and 5% among children. The prevalence of both infections increased with age. Concordant infections were found in 74.5% of adults (kappa=0.2) versus 2% of children (kappa=0.05). During the 9-year study period, the incidence of H. pylori infection was 1.1% among adults and 2% among children. The seroprevalence of HAV remained constant. The disparity between the increase in prevalence of H. pylori and HAV infection with age is likely associated with improvements in hygienic practices. The discordance between the presence of the infections among younger persons is evidence against a common source and/or vehicle for transmission.
Assuntos
Infecções por Helicobacter/epidemiologia , Helicobacter pylori , Vírus da Hepatite A Humana , Hepatite A/epidemiologia , Adolescente , Adulto , Fatores Etários , Criança , Estudos de Coortes , Estudos Transversais , Infecções por Helicobacter/complicações , Infecções por Helicobacter/imunologia , Hepatite A/complicações , Hepatite A/imunologia , Humanos , Japão/epidemiologia , Estudos Soroepidemiológicos , Testes Sorológicos , Fatores Sexuais , Fatores SocioeconômicosRESUMO
BACKGROUND: The purpose of this study was to examine the relationship between the production of secreted fibrinogen and the synthesis of gamma-chain mRNA. METHODS: We transfected a gamma-chain expression vector into Chinese hamster ovary cells already expressing both Aalpha- and Bbeta-chains of fibrinogen and measured fibrinogen output concentrations by ELISA. We quantified both gamma-chain and Bbeta-chain mRNA concentrations using the recently developed TaqMan fluorogenic detection system. RESULTS: The concentration of secreted fibrinogen into the media positively correlated with the amount of fibrinogen contained in the cell lysates. Additionally, quantitative mRNA assays revealed that the fibrinogen concentration in the cell lysates correlated well with the concentration of gamma-chain mRNA (r=0.7077, p<0.01) but not with the concentration of Bbeta-chain mRNA (r=0.0224, NS). CONCLUSIONS: These results demonstrate that the amount of recombinant fibrinogen produced in cells transfected with the gamma-chain vector, also expressing normal Aalpha- and Bbeta-chains, is dependent on the transcription of gamma-chain mRNA. Namely, in this recombinant expression system using a two-step transfection procedure, gamma-chain synthesis is the rate-limiting factor for fibrinogen production. This quantitative method to measure mRNA may prove very useful for further in vivo analysis of fibrinogen gene transcription.
Assuntos
Fibrinogênio/biossíntese , Fibrinogênio/genética , RNA Mensageiro/biossíntese , Transcrição Gênica , Sequência de Aminoácidos , Animais , Células CHO , Extratos Celulares/química , Cricetinae , Meios de Cultivo Condicionados/metabolismo , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Fibrinogênio/análise , Fibrinogênio/metabolismo , Humanos , Subunidades Proteicas , RNA Mensageiro/análise , RNA Mensageiro/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Padrões de Referência , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Investigation of the interactions of nerve cells with human apolipoprotein E (apoE), beta-amyloid (Abeta), and their complex, which are known to be included in senile plaques, is necessary to clarify the functional role of apoE in the pathogenesis of Alzheimer's disease. Using flow cytometric analysis, we investigated the isoform-specific effects of apoE on the endocytosis of Abeta in cultured neuroblastoma cells. The level of internalized Abeta within the cells was dependent on the culture time and the kind of apoE isoform present. Both apoE3 and apoE4 enhanced the internalization of Abeta; however, no difference was observed between their effects. The internalized Abeta was hardly catabolized at all in the presence of apoE4, while rapid clearance of Abeta was observed in the presence of apoE3. Unlike apoE3 and apoE4, apoE2 had no effect on Abeta clearance from the media. The isoform-specific effects of apoE on the endocytosis of Abeta may be implicated in the development of Alzheimer's disease, and if so, each isoform of apoE would induce a different incidence of that disease.