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Biomedical subjects

S Hirose

Publications and source records attributed to S Hirose.

At least 451 records · Page 25Linked to original sources
PubMed1991

Selective removal of anti-DNA and anticardiolipin antibodies by adsorbent plasmapheresis using dextran sulfate columns in patients with systemic lupus erythematosus.

The effect of adsorbent plasmapheresis using dextran sulfate columns on anti-DNA and/or anticardiolipin antibodies (aCL) in 6 patients with systemic lupus erythematosus (SLE) was studied by multicenter clinical trials. The titers of anti-DNA (RI assay), IgG anti-dsDNA (ELISA), IgG and/or IgM anti-ssDNA, (ELISA), and IgG aCL (ELISA) significantly decreased or normalized after 4 treatments of plasmapheresis during a 2 to 4 week period. A patient with SLE with recurrent abortion and aCL who was successfully treated by adsorbent plasmapheresis is reported. We expect that adsorbent plasmapheresis will be an influential treatment for patients with not only SLE but aCL syndrome.

Adsorption↗
PubMed1990

Human adrenal tumor cell line SW-13 contains a natriuretic peptide receptor system that responds preferentially to ANP among various natriuretic peptides.

A new type of ANP receptor system which clearly distinguishes natriuretic peptides A and B (ANP and BNP) has been identified in the human adrenal tumor cell line SW-13 and characterized. SW-13 cells responded to nanomolar concentrations of ANP with large increases in cGMP levels but in the case of BNP, much higher concentrations were required to produce the same extent of response. This property is unique since the 140-kDa ANP receptors so far characterized do not discriminate between ANP and BNP. For comparison, various natriuretic peptide receptors were also re-characterized using the recently identified CNP.

Adrenal Gland Neoplasms↗
PubMed1990

Identification and purification of a Bombyx mori homologue of FTZ-F1.

Extracts from embryos and from posterior and middle silk glands of the silkworm, Bombyx mori contain a sequence specific DNA binding factor termed BmFTZ-F1. The factor binds to the recognition site of FTZ-F1, a positive regulator of the fushi tarazu gene in Drosophila melanogaster. BmFTZ-F1 and FTZ-F1 share the same methylation interference patterns, the same chromatographic behaviors and similar protease digestion profiles. Anti-FTZ-F1 cross reacts with BmFTZ-F1. These results indicate that BmFTZ-F1 is a B. mori homologue of FTZ-F1. The mobility of the factor-DNA complex formed in the silk gland extract changes depending on the developmental stages. Purification of BmFTZF1 to an almost homogeneous state reveals that the factor is a 73 kd protein.

Animals↗
PubMed1990

Solubilization of endothelin receptors from bovine lung plasma membranes in a non-aggregated state and estimation of their minimal functional sizes.

Bovine lung endothelin receptors were solubilized in a non-aggregated state and characterized in terms of their minimal functional size and chemical nature of the ligand-binding site. A variety of detergents and their combinations were tested for their efficiency to solubilize endothelin receptors from bovine lung plasma membranes, and a combination of 0.4% digitonin and 0.25% CHAPS was found to be very effective in obtaining highly dispersed receptor solution. Gel filtration of the CHAPS/digitonin-solubilized receptors revealed the presence of two receptor species eluting at the positions corresponding to 34 and 52 kDa. These values were in good agreement with those estimated by affinity labeling and SDS-polyacrylamide gel electrophoresis, establishing that they represent minimal functional units. Chemical modification of ligand-occupied and free receptors with p-chloromercuriphenylsulfonic acid revealed that both 34 and 52 kDa receptors have SH group(s) essential for the receptor activity in their ligand-binding sites.

Affinity Labels↗
PubMed1990

A sandwich type enzyme-linked immunosorbent assay for proliferating cell nuclear antigen (PCNA)/cyclin using monoclonal antibodies.

Three hybridomas producing monoclonal antibodies to proliferating cell nuclear antigen (PCNA)/cyclin were newly derived. The specificity of established monoclonal antibodies to PCNA, TOB7, TO17 and TO30, was compared with that of the previously reported PCNA-specific monoclonal antibodies, 19A2 and 19F4. TOB7, TO17 and TO30 reacted with purified PCNA in an enzyme-linked immunosorbent assay (ELISA). A 34 kDa PCNA polypeptide was noted by immunoblotting, the same polypeptide as recognized by 19A2 and 19F4. Epitopes recognized by all those monoclonal antibodies to PCNA were analyzed by competitive inhibition tests using ELISA. The results of those experiments suggested that the epitope recognized by TO17 and TO30 was almost identical to that of 19A2 and closely related to that of 19F4, but different from that of TOB7. Based on those results, a sandwich type ELISA for detection of PCNA was developed using TO17 and TOB7. This system was applied to measure the concentration of PCNA in normal peripheral blood lymphocytes (PBL) before and after stimulation by phytohemagglutinin (PHA), and in tissue extracts from rabbit kidney and thymus (RKE and RTE, respectively). Before mitogenic stimulation, the PCNA concentration in PBL extracts was less than 6 ng/ml (cell concentration 2.5 x 10(7)/ml), but at 48 h after PHA stimulation, when more than 70% of cells were in S phase, its concentration became 1280 ng/ml. RTE which contained many proliferative lymphocytes had much higher concentration of PCNA than RKE. Those results suggested that sandwich type ELISA using TO17 and TOB7 was useful as the quantitative assay to detect blast-transformation and proliferation of cells in vivo.

Antibodies, Monoclonal↗
PubMed1990

Lymphokine-regulated differential expression of mRNA for p75kDa-IL-2R and p55kDa-IL-2R in a cloned B lymphoma line (BLC1-CL-3 cells).

IL-5 renders BCL1-CL-3 (CL-3) cells responsive to IL-2 by increasing the number of high affinity IL-2R, whereas IL-4 prohibits such action of IL-5 to prepare CL-3 cells responsive to IL-2. Here we have found that genes for p75kDa-IL-2R and p55kDa-IL-2R are differentially regulated by IL-4 and IL-5. Nonstimulated CL-3 cells constitutively express mRNA for p75kDa-IL-2R and p55kDa-IL-2R. IL-5 stimulation principally augments the expression of p75kDa-IL-2R mRNA (4- to 8-fold), although modestly increasing the expression of p55kDa-IL-2R mRNA. Kinetic studies have revealed a maximal increase in p75kDa-IL-2R mRNA expression at 12 h and a decline thereafter, substantiating our previous kinetic study of the expression of high affinity IL-2R after the IL-5 stimulation. By contrast, IL-4 stimulation modestly increases the expression of p75kDa-IL-2R mRNA, whereas markedly reducing the expression of p55kDa-IL-2R mRNA, irrespective of whether CL-3 cells were stimulated with IL-4 alone or together with IL-5 and IL-2. Moreover, addition of IL-4 into the culture containing IL-5 and IL-2 causes striking reduction in the level of J-chain mRNA, which otherwise is markedly induced by stimulation with IL-5 and IL-2. These results clearly illustrate the differential regulation of p75kDa- and p55kDa-IL-2R-gene expression by IL-5 and IL-4, and reinforce our notion that increased expression of high affinity IL-2R induced by IL-5 is responsible for the IL-2 competent state, and decreased expression of p55kDa-IL-2R by IL-4 is responsible for IL-2 unresponsive state.

Animals↗
PubMed1990

High-performance liquid chromatography with chemiluminescence detection of serum levels of pre-column derivatized fluoropyrimidine compounds.

7-(Diethylamino)-3-[4-[iodoacetyl)amino)phenyl]-4-methylcoumarin (DCIA) and 4-(bromomethyl)-7-methoxycoumarin have been evaluated as fluoropyrimidine-derivatizing agents to be detected using peroxyoxalate chemiluminescence with high-performance liquid chromatography. The derivatization procedure required only one step. No chemiluminescence was observed from the bromo derivatives, and the detection limits of fluoropyrimidine compounds derivatized with the iodo compound and detected with peroxyoxalate chemiluminescence were in the low femtomole range.

Chromatography, High Pressure Liquid↗
PubMed1990

Primary structure of human placental 5'-nucleotidase and identification of the glycolipid anchor in the mature form.

A cDNA was cloned coding for human placental 5'-nucleotidase. The 3547-bp cDNA contains an open reading frame that encodes a 574-residue polypeptide with calculated size of 63 375 Da. The NH2-terminal 26 residues comprise a signal peptide, which is followed by the NH2-terminal sequence of the purified protein. four potential N-linked glycosylation sites are found in the molecule, accounting for a larger mass of the mature form (71 kDa). The predicted structure contains a hydrophobic amino acid sequence at the COOH terminus, a possible signal for the post-translational modification by glycophospholipid. To confirm this possibility, we tried to isolate and characterize the membrane-anchoring domain of 5'-nucleotidase. BrCN-cleaved fragments of the protein were extracted with hexane and subjected to HPLC, resulting in purification of a single component of 2.3 kDa. Chemical analyses revealed that the purified fragment contains the tetradecapeptide Lys-Val-Ile-Tyr-Pro-Ala-Val-Glu-Gly-Arg-Ile-Lys-Phe-Ser, ethanolamine, glucosamine, mannose, inositol, palmitic acid, and stearic acid. The peptide sequence determined is identified at positions 510-523 in the primary structure deduced from the cDNA sequence, which predicts a further extension to position 548, containing the hydrophobic amino acid sequence. Thus, it is concluded that the mature 5'-nucleotidase lacks the predicted COOH-terminal peptide extension (524-548), which has been replaced by the glycophospholipid functioning as the membrane anchor of 5'-nucleotidase.

5'-Nucleotidase↗
PubMed1990

Determination of purine nucleosides and their bases by high-performance liquid chromatography using co-immobilized enzyme reactors.

A selective and sensitive assay of inosine, guanosine, hypoxanthine, guanine and xanthine by high-performance liquid chromatography with immobilized enzyme reactors was developed. The separation was achieved on a Capcell Pak C18 column (15 cm x 0.46 cm I.D.) with a mobile phase of 0.1 M phosphate buffer (pH 8.0) containing 7 mM sodium 1-hexanesulphonate and 0.1 mM p-hydroxyphenylacetic acid. The fluorimetric detection of hydrogen peroxide using immobilized peroxidase and p-hydroxyphenylacetic acid was applied to the assay of these compounds, which were oxidized to yield hydrogen peroxide in the presence of immobilized enzyme (purine nucleoside phosphorylase, guanase and xanthine oxidase). Enzyme reactions occurred sufficiently without post-column addition of reagents. Enzymes that catalysed the conversion of purine compounds were co-immobilized on aminopropyl controlled-pore glass packed in stainless-steel tubing. The detection limits were 30-200 pg per injection.

Chromatography, High Pressure Liquid↗
PubMed1990

Primary structure of rat liver 5'-nucleotidase deduced from the cDNA. Presence of the COOH-terminal hydrophobic domain for possible post-translational modification by glycophospholipid.

Rat liver 5'-nucleotidase was purified from a crude microsomal fraction, and its molecular mass was estimated to be 73 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified protein was subjected to cleavage with CNBr or lysyl endopeptidase, and the resulting 21 peptides as well as the NH2 terminus of the native protein were sequenced by Edman degradation. For further information on the molecular structure, we constructed a lambda gt11 liver cDNA library and isolated two cDNA clones for 5'-nucleotidase, lambda cNTP6 and lambda cNT34. The 3.2-kilobase cDNA insert of lambda cNTP6 contains an open reading frame that encodes a 576-residue polypeptide with a calculated size of 63,965 Da, which is in reasonable agreement with that of 5'-nucleotidase (62 kDa) immunoprecipitated from cell-free translation products. The NH2-terminal 28 residues comprise a signal peptide, which is followed by the NH2-terminal sequence of the purified protein. The predicted structure contains all the other peptide sequences determined by Edman degradation. Five potential N-linked glycosylation sites are found in the molecule, accounting for the difference in mass between the precursor and mature forms. Another characteristic feature is that the primary structure contains a highly hydrophobic amino acid sequence at the COOH terminus, a possible signal for the post-translational modification by glycophospholipid. In fact, labeling experiments of rat hepatocytes demonstrated that 3H-labeled compounds such as ethanolamine, myo-inositol, and palmitic acid, components of the glycolipid anchor, were incorporated into 5'-nucleotidase. Phosphatidylinositol-specific phospholipase C released 5'-nucleotidase from the cell surface, and the released protein no longer contained the radioactivity of [3H]palmitic acid incorporated.

5'-Nucleotidase↗
PubMed1990

IL-5 up-regulates but IL-4 down-regulates IL-2R expression on a cloned B lymphoma line.

Both IL-4 and IL-5 demonstrate B cell growth activity. IL-5 can render a cloned neoplastic B cell line, BCL1-CL-3 cells, responsive to IL-2, whereas IL-4 has no such activity. The response to IL-5 and IL-2 proceeds in two phases: the first phase which clearly depends upon IL-5 is the obvious increase in number of high affinity IL-2R (3.1-fold) with modest increase of low affinity IL-2R (1.2-fold) and gain of the ability of facilitated IL-2-binding and internalization of IL-2, and the second phase which is induced by IL-2 in the IL-5-stimulated CL-3 cells comprises the striking increase of low affinity IL-2R (8.5-fold). Kinetic study has revealed that high affinity IL-2R expressed on CL-3 cells begins to increase at 6 h and reaches to maximum at 12 h after stimulation with IL-5 or IL-5 plus IL-2, whereas low affinity IL-2R expression increases at 18 h and becomes maximal at 24 h after stimulation of CL-3 cells with IL-5 and IL-2. However, in the presence of IL-4, IL-5 cannot induce an increase in number of high affinity IL-2R on CL-3 cells. Thus, CL-3 cells stimulated with the mixture of IL-5 and IL-4 cannot respond to IL-2, and fail to show up-regulated expression of low affinity IL-2R. IL-4 also has a capacity to modestly interfere with the action of IL-2 to up-regulate low affinity IL-2R expression on IL-5-pretreated CL-3 cells. Thus, this monoclonal B cell system provides an excellent model system to define the roles of IL-5 and IL-4 involved in the B cell differentiation and to characterize the properties of B cells competent to IL-2 stimulation and the signal transduction mechanism which operates through IL-2/IL-2R system.

Animals↗
PubMed1990

Core promoter of the mouse myelin basic protein gene governs brain-specific transcription in vitro.

The core promoter of the mouse myelin basic protein (MBP) gene from -36 to +12 was preferentially transcribed in brain nuclear extracts. Both the TATA at -34 and downstream elements to +12 were required for efficient, accurate and brain-specific transcription. From brain and liver nuclear extracts, we have partially purified the general transcription factor TFIID. The partially purified fractions contained TATA element binding factors of the MBP promoter as well as adenovirus major late promoter (MLP). The tissue-derived TFIID was functionally exchangeable for the HeLa TFIID, and directed transcription from the MLP. Surprisingly, the brain TFIID activated transcription from the MBP core promoter while the liver TFIID did to a much lesser extent. Exchange of the TATA-containing short DNA stretch to the MBP core promoter for a corresponding region of the mouse albumin promoter or MLP abolished the brain specificity. We found that several tissue-specific promoters other than MBP, such as mouse neurofilament and human alpha-1-antitrypsin promoters were also transcribed much more efficiently by the brain and liver TFIID, respectively. We suggest that different tissues contain functionally non-equivalent TFIID or TFIID-like activities.

Animals↗
PubMed1990

Activated peripheral blood mononuclear cells detected by murine monoclonal antibodies to proliferating cell nuclear antigen in active lupus patients.

Hybridoma producing monoclonal antibody to proliferating cell nuclear antigen (PCNA/cyclin) (TOB7, IgG1 kappa) was newly established. Using TOB7, PCNA was detected in the peripheral blood mononuclear cells (PBMC) in patients with systemic lupus erythematosus (SLE). Forty-four of 58 patients with SLE had PBMC expressing PCNA. The percentage of PCNA-positive PBMC in patients with SLE was 0-20% (mean: 2.63%) which was significantly higher (P less than 0.01) compared with normal controls (mean: 0.18%), patients with rheumatoid arthritis (mean: 0.83%), and patients with mixed connective tissue disease (mean: 0.38%). Patients with high numbers of PCNA positive PBMC tended to complicate pulmonary disorders (P less than 0.005), especially pulmonary fibrosis (P less than 0.005). In addition, the percentage of PCNA-positive cells in SLE patients correlated with the disease activity (r = 0.45, P less than 0.01). The lymphocyte subsets of PCNA-positive PBMC were examined, and most of those cells belonged to CD4- or CD8-positive T-cell populations in three lupus patients. Our findings indicate that PCNA-positive activated PBMC are present in SLE patients and the percentage of PCNA-positive PBMC may be used as an indicator of disease activity.

Adolescent↗
PubMed1990

HLA-DP+ T cells and deficient interleukin-2 production in patients with systemic lupus erythematosus.

In patients with systemic lupus erythematosus (SLE), frequency of the T cells positive for HLA-DP, one of the major histocompatibility complex (MHC) class II molecules, was markedly increased in peripheral blood lymphocytes (PBL), in association with an increase in the amount of specific cytoplasmic transcript of the HLA-DP gene segment. Cell cycle analysis showed that HLA-DP is an early activation marker of T cells and that the high ratios of HLA-DP+ T cells from SLE patients are associated with high frequency of T cells at early activation phases, mainly of G1A. Initial high ratios of HLA-DP+ T cells decreased to a great extent during 4 days of in vitro culture, in the absence of mitogens. This event was associated with decreases in the amount of HLA-DP transcript and the disappearance of activated T cells. Studies on the interleukin 2 (IL-2) production of T cells from patients with SLE demonstrated that while the PBL rich in HLA-DP+ T cells show a markedly low production of IL-2, preculture of these PBL restores the ability to produce IL-2. Thus, it appears that the T cells in patients with SLE are essentially intact with regard to the capacity to produce IL-2 and that T cell activation events continuously occurring in SLE patients are related to a deficiency in IL-2 production. The possible underlying mechanisms are discussed.

Adolescent↗
PubMed1990

Comparative experimental study of antiepileptics--regional specificity of antiepileptic action of carbamazepine, phenobarbital and valproate sodium.

The antiepileptic action of carbamazepine (CBZ), phenobarbital (PB), and valproate sodium (VPA) was examined in 109 adult male rabbits during steady state serum levels produced on the basis of the pharmacokinetics of each drug. The duration and discharge pattern of electrically induced focal and secondarily generalized after-discharges (ADs) in the motor, visual cortices and hippocampus were compared before and after administration of doses of each drug. A CBZ level of 3-30 micrograms/ml shortened the duration of both neocortical and hippocampal ADs. CBZ at levels above 5 micrograms/ml suppressed the spread in the secondarily generalized ADs originating from the neocortical areas, but not that originating from the hippocampus. PB levels of 10-80 micrograms/ml shortened the duration of both neocortical ADs, especially motor ADs, whereas they had little or no effect on the hippocampal ADs. VPA showed inhibitory effects on both the neocortical and hippocampal AD durations only at high levels of 400-1,100 micrograms/ml. However, the inhibitory effects were more marked on the latter than the former, and further, the effects were enhanced as time elapsed. These results suggest that the action of each drug on focal seizures is region-specific.

Animals↗
PubMed1990

Purification of a DNA supercoiling factor from the posterior silk gland of Bombyx mori.

A protein factor with an estimated molecular mass of 50 kDa has been purified to homogeneity from the silk gland of Bombyx mori. In the presence of a molar excess of this factor and eukaryotic DNA topoisomerase II, relaxed circular DNA is converted to the negatively supercoiled form. Eukaryotic DNA topoisomerase I cannot substitute for eukaryotic DNA topoisomerase II in the supercoiling reaction. The reaction is dependent on ATP and is inhibited by VP-16, a specific inhibitor of eukaryotic DNA topoisomerase II. When DNA topoisomerase I is subsequently added to the supercoiling reaction mixture, the supercoiled DNA becomes relaxed. These results suggest that when both the 50-kDa protein and eukaryotic DNA topoisomerase II are present in excess, unconstrained negative supercoils are introduced into DNA.

Animals↗