A hidden mitogenic activity of Vitis (grape) extract for human lymphocytes in the presence of monocytes or serum.
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Blood↗
Biomedical subjects
H Matsui
Publications and source records attributed to H Matsui.
Substrate specificity and subsite affinities of rabbit liver acid alpha-glucosidase.
The substrate specificity of rabbit liver acid alpha-glucosidase was investigated. The enzyme showed a wide specificity for various substrates, and hydrolyzed alpha-glucans such as glycogen and soluble starch. The k0 values (s-1) for maltose, kojibiose, nigerose, isomaltose, phenyl alpha-glucoside, panose, phenyl alpha-maltoside, soluble starch, beta-limit dextrin, amylopectin, shellfish glycogen, and rabbit liver glycogen were estimated to be 94.8, 18.8, 143, 3.6, 11.8, 27.8, 115, 99.2, 155, 83.5, 126, and 108, and the Km values (concentration of non-reducing terminal) for these substrates were 2.1, 1.8, 7.5, 36, 5.4, 1.9, 1.2, 0.90, 9.1, 1.0, 16, and 13 mM, respectively. Isomaltose and phenyl alpha-glucoside were unfavorable as substrates. The acid alpha-glucosidase is characterized by a relatively high activity toward glycogen. The k0 values (s-1) for maltotriose, -tetraose, -pentaose, -hexaose, -heptaose, and -octaose, and maltodextrin (n = 17) were 140, 140, 131, 132, 134, 132, and 74.3, and the Km values, 2.1, 1.8, 1.9, 3.4, 5.0, 4.9, 4.9 and 2.6 mM, respectively. Based on the rate parameters for the series of maltooligosaccharides, the subsite affinities (Ais) in the active site were evaluated as 0.54 (A1), 5.34 (A2), and 0.34 (A3) kcal/mol for subsites 1, 2, and 3, respectively. These three subsites were considered to be predominantly responsible for the binding of substrates to the active site.
Animals↗
Ceramic vertebral body replacement for metastatic spine tumors.
A modular ceramic spacer system was developed and used for vertebral body replacement in 21 patients with metastatic spine tumor in cervical (n = 10 cases), thoracic (n = 9), and lumbar spines (n = 2). All cases of cervical and lumbar spinal tumors underwent simultaneous use of a ceramic spacer and plate-screw fixation, and eight patients received additional posterior interventions in the thoracic spine. Postoperative radiation therapy was performed in all cases except two, which involved prostatic carcinoma. Complete or moderate pain relief was obtained in 19 patients, and 17 patients were able to walk after surgery. The average bed rest period was 11 days. Radiologically, a clear zone at the spacer-vertebral body interfaces and migration of the spacer was observed in four cases and one case, respectively. However, satisfactory pain relief and stabilization of the spine was obtained in an average follow-up time of 22.3 months. In all four autopsy specimens studied, the ceramic spacer showed firm connection with the vertebral body. This modular ceramic spacer system facilitates a relatively easy procedure, maintenance of spinal stability, early ambulation, and no interference with local radiation therapy.
Adult↗
Molecular cloning and characterization of a murine AIDS virus-related endogenous transcript expressed in C57BL/6 mice.
The murine AIDS (MAIDS) virus has a unique sequence in the gag p12 region, which could be responsible for MAIDS development. RNA preparations from the spleens of normal uninfected C57BL/6 mice contain a transcript hybridizing with this sequence. Levels of the transcript in the kidney of C57BL/6 mice were higher than in the spleen, liver or thymus. Although BALB/c, NFS, DBA/2 and SL murine strains also contained genomic sequences hybridizing with the MAIDS virus-specific probe, no transcript hybridizing with the probe was detected in these strains of mice. The cDNAs carrying the transcript expressed in C57BL/6 mice were molecularly cloned. The complete nucleotide sequence of the clone indicates that the transcript is one of the endogenous murine leukaemia virus-related sequences containing large deletions from the R and U5 regions of the 5' long terminal repeat (LTR) to gag p15, from the C-terminal region of pol p40 (integrase) to the N-terminal region of env p15E, and many short deletions in the 3' LTR U3 region. The nucleotide sequence in the gag p12 region of the transcript was closely similar to that of the MAIDS virus, but the amino acid sequence was less similar because of frameshifting, even when translated. As the MAIDS virus was isolated from C57BL/6 mice with radiation-induced leukaemia, this transcript may be the progenitor of the MAIDS virus. To determine whether the gag p12 region of the transcript contains a functional sequence, a recombinant virus was generated by replacing the gag p12 region of a replication-competent BM5eco virus with that of the endogenous transcript. The recombinant virus was replication-competent, and the p12 region of the transcript retained the functional sequence present in the BM5eco virus.
Amino Acid Sequence↗
Virulence region of plasmid pNL2001 of Salmonella enteritidis.
The virulence region of the Salmonella enteritidis 55 kb plasmid pNL2001 was identified by Tn1-insertion mutagenesis, DNA hybridization studies, and Western blot analysis of proteins encoded in the virulence region of the plasmid. DNA hybridization studies showed that the pNL2001 plasmid contained a 6.4 kb SalI-EcoRI fragment homologous to the 6.4 kb SalI-EcoRI Salmonella plasmid virulence (spv) region of the S. choleraesuis 50 kb plasmid (pKDSC50). One of the 247 Tn1-insertion mutants of S. enteritidis, designated strain TA19, showed a reduced mouse lethality, and the Tn1-insertion of strain TA19 was located within this homologous 6.4 kb region, suggesting that the 6.4 kb SalI-EcoRI fragment of pNL2001 contained the spv region. Two contiguous SalI-EcoRI fragment were cloned into the expression vectors. By the 6.4 kb SalI-EcoRI fragment were cloned into the expression vectors. By Western blot analysis using four Spv peptide antisera, each specific for individual proteins encoded in the spvR, spvA, spvB and spvC genes of pKDSC50, four proteins encoded in the 6.4 kb SalI-EcoRI fragment of pNL2001 were identified. Protein SpvR with an apparent molecular mass of 32 kDa was produced from the 2.3 kb SalI-EcoRI fragment, and protein SpvA, SpvB and SpvC with apparent molecular masses of 32, 70 and 29 kDa, respectively, were produced from the 4.1 kb EcoRI-EcoRI fragment. From the 4.1 kb EcoRI::Tn1 fragment of the TA19 plasmid, proteins SpvA and SpvB were expressed, but not SpvC. It was therefore suggested that the spvC gene may contribute to the expression of virulence of S. enteritidis. Furthermore, the nucleotide sequence of the 6.4 kb SalI-EcoRI fragment encoding these four proteins was determined. Four open reading frames which encoded the four proteins with deduced molecular masses of 33,906, 28,200, 65,349 and 27,646 Da were detected. Deduced amino acid sequences of each protein showed a high degree of identity to corresponding sequences in the virulence region of S. choleraesuis, S. dublin and S. typhimurium virulence plasmids. Therefore, we confirmed that the virulence plasmids of Salmonellae including S. enteritidis share the highly conserved region responsible for virulence.
Amino Acid Sequence↗
Effects of epidermal growth factor on invasiveness through the extracellular matrix in high- and low-metastatic clones of RCT sarcoma in vitro.
We investigated the invasiveness of tumor cells through the extracellular matrix and the influence of epidermal growth factor (EGF) on tumor cell invasion using in vitro systems in high-[RCT(+)] and low-metastatic [RCT(-)] clones established from poorly differentiated murine RCT sarcoma in C3H/He mice. In the invasion assay using a filter coated with reconstituted basement membrane (Matrigel) in a Boyden chamber, RCT(+) cells were more invasive than RCT(-) cells. The attachment of RCT(+) cells to extracellular matrix components and the degradation of type IV collagen by the cells were significantly greater than with RCT(-) cells. However, there was no significant difference in the migration of cells to the extracellular matrix components between cultured RCT(+) and RCT(-) cells. These findings suggested that the different invasiveness of these clone cells was associated with the difference in the ability of attachment to and degradation of the matrix. The level of laminin receptor expression in RCT(+) cells was about four-fold that in RCT(-) cells and laminin stimulated the type IV collagenolytic activity of RCT(+) cells, suggesting that RCT(+) cell attachment to laminin via laminin receptor on the cell surface induced the production of type IV collagenase by the tumor cells. EGF did not affect the invasiveness of RCT(-) cells. In RCT(+) cells, EGF stimulated the invasiveness through Matrigel, the attachment to extracellular matrix components and the degradation of type IV collagen through high-affinity EGF receptors (EGFR), with Kd of pM order, while the migration to the matrix was not influenced by EGF. These findings suggest that the stimulatory effect of EGF on invasion is related to the acceleration of cell adhesion, and the degradative cascade of the extracellular matrix and high-affinity EGFRs play an important role in the effect of EGF on in vitro invasiveness in this tumor.
Animals↗
Serum levels of laminin, type IV collagen and type III procollagen peptide as markers for detection of metastasis.
We investigated the clinical usefulness of serum laminin, type IV collagen and type III procollagen peptide (PIIIP) as markers for detection of metastasis in patients with primary or metastatic bone and soft part tumors. The subjects consisted of 28 patients with metastatic bone tumors, 18 with primary bone tumors (benign; 10, malignant; 8), 22 with primary soft part tumors (benign; 12, malignant; 10), 18 with cancer without metastasis (as controls to metastatic bone tumor) and 60 healthy controls. Elevated levels of serum laminin, type IV collagen and PIIIP were not associated with any specific histological subtype, tumor size or location, and were clearly related to evidence of metastasis. Mean serum concentrations of laminin, type IV collagen and PIIIP were significantly higher in patients with metastasis than in patients without metastasis. Positive correlations were observed among serum laminin, type IV collagen and PIIIP levels in tumor patients. The sensitivity values for laminin, type IV collagen and PIIIP in detecting metastasis were 83.7%, 83.3% and 80.5%, respectively, with specificity of 90.0%, 86.1% and 86.1%. When two of the three markers were evaluated in identical blood samples, combined sensitivity and specificity values exhibited further increases as compared to the sensitivity and specificity of each marker. The use of all three markers led to the best combined sensitivity and specificity. These findings suggest that the combination of these markers would be a valuable screening test in predicting metastasis.
Analysis of Variance↗
Regulation of spvR gene expression of Salmonella virulence plasmid pKDSC50 in Salmonella choleraesuis serovar Choleraesuis.
The expression regulation of spvR, a regulatory gene on the virulence plasmid (pKDSC50) of Salmonella choleraesuis serovar Choleraesuis, was investigated by spvR-lacZ translational fusion. The spvR gene was found to be positively regulated by its own product, the SpvR protein, and this unusual positive autoregulation was repressed by the products of spvA and spvB, virulence-associated genes present downstream from the spvR gene. Amino acid sequence analysis revealed that the amino-terminal region of SpvB had homology with the CatM repressor protein of Acinetobacter calcoaceticus, which belongs to the MetR/LysR protein family. On the other hand, the sigma factor RpoS was required for expression of the spvR gene in the stationary phase of bacterial growth. The SpvR protein was also necessary for self-activation, suggesting that an RNA polymerase holoenzyme containing RpoS requires SpvR protein in order to recognize the spvR promoter.
Base Sequence↗
Molecular cloning and expression of an isomalto-dextranase gene from Arthrobacter globiformis T6.
The gene encoding an extracellular isomalto-dextranase, designated imd, was isolated from the chromosomal DNA of Arthrobacter globiformis T6 and cloned and expressed in Escherichia coli. A single open reading frame consisting of 1,926 base pairs that encoded a polypeptide composed of a signal peptide of 39 amino acids and a mature protein of 602 amino acids (M(r), 65,900) was found. The primary structure had no significant homology with the structures of any other reported carbohydrases, including two other dextranases. Transformed E. coli cells carrying the 2.3-kb fragment overproduced isomalto-dextranase into the periplasmic space under control of the promoter of the imd gene itself.
Amino Acid Sequence↗
Adenovirus pulmonary infections identified by PCR and in situ hybridisation in bone marrow transplant recipients.
AIMS: To investigate adenovirus pulmonary infections in bone marrow transplant (BMT) recipients. METHODS: Formalin fixed, paraffin wax embedded lung tissue was examined from 13 necropsy cases after BMT using PCR and in situ hybridisation to detect adenovirus DNA. The E1A region of the adenoviral genome was targeted for PCR. In situ hybridisation was performed only in the PCR positive cases. RESULTS: Of the 13 lung specimens analysed, nine cases were negative for adenoviral nucleic acid. Four (30%) PCR and two (15%) in situ hybridisation positive cases were found. In some of the patients there were clinical and pathological indications that some diseases might be associated with adenovirus infection--haemorrhagic cystitis (three cases); necrotising pneumonia (one case). In necrotising pneumonia in which no pathogenic agents had been shown by conventional histological study, the in situ hybridisation technique showed positive staining for adenovirus. In a patient who died of renal failure caused by adenovirus nephritis, both PCR and in situ hybridisation were positive in the lung as well as in the kidney, although no histological change was found. Two PCR positive cases lacked positive sites for adenovirus by in situ hybridisation. CONCLUSIONS: The combination of PCR and in situ hybridisation could be useful for diagnosing adenovirus infection of the lung in BMT recipients. These results provide a basis for exploring further the clinical use of PCR and in situ hybridisation to diagnose adenovirus infection.
Adenovirus Infections, Human↗
Sarcoplasmic reticulum gene expression in cardiac hypertrophy and heart failure.
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Animals↗
Changes in annexin I and II levels during the postnatal development of rat pancreatic islets.
The expression patterns and the dynamic changes in content of both annexin I and annexin II in the rat pancreatic islets during postnatal development were investigated by both western blot analysis and immunohistochemistry. Immunohistochemical methods clearly demonstrated the presence of annexins I and II exclusively in pancreatic islets, while exocrine tissues were not stained by anti-annexin antibodies. Pancreatic islets were diffusely stained with no specific differences in distribution between different cell types. The expression of annexin I in pancreatic islets gradually increased with postnatal development. A developmental study of annexins I and II by western blot analysis essentially supported the results obtained by immunohistochemistry. In addition, the increasing expression of two protein tyrosine kinases, epidermal growth factor-receptor/kinase and pp60src, which phosphorylate annexin I and annexin II, respectively, and of protein kinase C, which phosphorylates both proteins, was also shown during postnatal development in rat pancreatic islets. Thus, a relationship between the expression of annexins I and II and the maturation of islet cell function is suggested.
Actins↗
Chemical synthesis of the gene for microbial transglutaminase from Streptoverticillium and its expression in Escherichia coli.
The gene coding for microbial transglutaminase (TGase) from Streptoverticillium, which consists of 331 amino acids, was chemically synthesized. The codons have been substituted for those mainly favored in yeast. Our strategy involved the construction of the TGase gene in five sections (54 oligomers) that contained unique restriction enzyme sites at both ends, which could readily be ligated to form the full-length product. The chemically synthesized gene was inserted downstream from the ompA signal peptide of the E. coli expression vector, pIN-III-ompA, which carries lpp and lac promotors. The resultant plasmid directed the expression of TGase, with the activity being secreted mainly into the periplasmic space of E. coli. The induced gene product was identical with native TGase in size and in immunological properties, though the enzyme activity was low.
Amino Acid Sequence↗
Formaldehyde-fixed cells infected with pseudorabies virus conferred resistance against Aujeszky's disease.
Cells infected with Pseudorabies virus (PrV) and fixed with formaldehyde were evaluated as a vaccine against Aujeszky's disease. Mice and pigs inoculated with fixed cells showed no clinical signs. In a challenge test, all fixed cells conferred protection on mice and pigs. Although pigs showed slight symptoms, they were restored to normal health within a few days. These results demonstrate that PrV-infected cells fixed with formaldehyde function as a vaccine against Aujeszky's disease and serve as a novel type of vaccine which is easy to make, and is both safe and effective.
Animals↗
Production and characterization of an anti-chicken transferrin.
Several lines of recent evidence have suggested that transferrin plays a significant role in tissue interaction or morphogenesis at early stages of embryo development. In the present study, an anti-chicken transferrin antibody was produced and its basic characteristics were clarified as a basis for use in further studies. An antiserum termed Toraji 3 was raised against chicken transferrin and purified into IgG and ligand-affinity-purified fractions. These three preparations of the antibody gave an intense immunohistochemical signal in visceral yolk sac and developing liver, both of which are known to be major producers of transferrin in early development. In immunoblot analysis, these three preparations detected 70-kDa transferrin, whereas the ligand-affinity-purified preparation showed higher specificity. It was also demonstrated by enzyme-linked immunosorbent assay and immunoblotting that Toraji 3 antibody bound preferentially to chicken transferrin and showed a negligible binding to human transferrin.
Animals↗
[Diffuse aspiration bronchiolitis (DAB) produced in animals by repeated HCl microaspiration].
We recently reported that diffuse aspiration bronchiolitis (DAB) was detected in 1% of autopsied lungs of aged cases of pneumonia. We hypothesized whether repeated HCl micro-aspiration (RHMA) is involved in DAB and established an animal model by administering HCl intratracheally to rats every two days for 2 weeks. Saline was given to control animals in the same fashion. Then, we performed bronchio-alveolar lavage (BAL) or excised lungs for histologic examination. There was no difference in BAL cell counts, TNF alpha-production, elastase-like activity or albumin levels between the HCl and control groups. Histologically, DAB-like findings were observed in the HCl-treated animals. These data suggest that RHMA might be involved in DAB, while neither TNF alpha-production nor elastase-like activity may play a significant role in inducing DAB.
Animals↗
Involvement of annexin I and annexin II in hepatocyte proliferation: can annexins I and II be markers for proliferative hepatocytes?
Annexin is the name of a new family of Ca(2+)-dependent membrane-binding proteins. Eleven types of its related proteins have been reported to date. Among those, annexin I and annexin II have been reported to possess many biological functions in vitro. Its actual role in vivo, however, is yet unknown. The involvement of annexin I and annexin II in the proliferation processes of hepatocytes was examined in the following aspects: (a) hepatocyte proliferation after carbon tetrachloride-induced liver damage, (b) hepatocyte regeneration after partial hepatectomy and (c) postnatal development of hepatocytes. These results showed collectively that annexin I and annexin II were increased in proliferative (or regenerative) hepatocytes, suggesting that both proteins play a certain role in the proliferation event. Furthermore, annexin I- and annexin II-positive hepatocytes always show a wider distribution than that of proliferating cell nuclear antigen or cytokeratin 7-positive hepatocytes, indicating that annexin I and annexin II may be useful markers for detecting not only actively proliferating hepatocytes but also hepatocytes in preproliferative and postproliferative stages.
Animals↗
Modulation of adrenergic receptors during regression of cardiac hypertrophy.
OBJECTIVE: To determine whether alpha 1- or beta-adrenergic receptors are altered during regression of cardiac hypertrophy produced by antihypertensive agents. DESIGN AND METHODS: Cardiac hypertrophy was induced in rats by aortic banding. After 6 weeks banding the rats were treated with an angiotensin converting enzyme (ACE) inhibitor (enalapril), an alpha 1-adrenergic antagonist (bunazosin) or a beta-adrenergic antagonist (propranolol) for 6 weeks to induce regression. The numbers of alpha 1- and beta-adrenergic receptors, haemodynamics, tissue noradrenaline content and tissue ACE activity were measured. RESULTS: Regression of cardiac hypertrophy occurred after treatment of aortic banded rats with a high dose of enalapril, bunazosin or propranolol, and was accompanied by a reduction in systolic blood pressure. The number of alpha 1- or beta-adrenergic receptors was unchanged by propranolol treatment, but the number of alpha 1-adrenergic receptors was increased in the hearts of rats treated with bunazosin. A low dose of enalapril (3 mg/kg body weight) caused regression of hypertrophy without a concomitant reduction in blood pressure, and decreased the number of alpha 1-adrenergic receptors. The dissociation constants for alpha 1- and beta-adrenergic receptors were not different among the experimental groups, and the positive derivatives of left ventricular pressure was unaltered in rats treated with a low dose of enalapril but was reduced by the other drugs. CONCLUSION: Of the three drugs tested, only the low dose of enalapril affected adrenergic receptors during regression of cardiac hypertrophy, causing a decrease in alpha 1-adrenergic receptor number without a reduction in blood pressure. This effect may be explained by non-haemodynamic actions of the ACE inhibitor enalapril, probably by modulation of peripheral sympathetic activity.
Angiotensin-Converting Enzyme Inhibitors↗