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Biomedical subjects

M Yamaguchi

Publications and source records attributed to M Yamaguchi.

At least 739 records · Page 41Linked to original sources
PubMed1996

Purification, characterization and cDNA cloning of a novel anticoagulant of the intrinsic pathway, (prolixin-S) from salivary glands of the blood sucking bug, Rhodnius prolixus.

The salivary glands of the blood sucking insect, Rhodnius prolixus, have an anticoagulant, prolixin-S, which was reported as a specific inhibitor of intrinsic coagulant pathway. Prolixin-S was purified from the salivary glands extract of Rhodnius prolixus by gel filtration and anion exchange HPLC by assaying prolongation of activated partial thromboplastin time (APTT). The isolated protein specifically inhibited factor IXa-catalyzed activation of factor X in the presence of Ca2+ and phospholipids irrespective of the presence or absence of factor VIIIa. The anticoagulant factor had red color and a specific absorbance peak at 402 nm and thus it was identified as a heme protein. A Rhodnius prolixus salivary gland cDNA library was prepared, screened with an antibody against prolixin-S and its complete cDNA sequence was determined. cDNA and deduced amino acid sequences showed that prolixin-S is a novel anticoagulant of 19,922 Da, which has no sequence homology with any other anticoagulant reported so far.

Amino Acid Sequence↗
PubMed1996

Effect of corn peptide on alcohol metabolism and plasma free amino acid concentrations in healthy men.

OBJECTIVES: To determine the influence of corn peptide on the reduction of the increase in blood ethanol level in healthy adult men after intake of alcohol at a dose of a 0.5 g per kg of body weight. SUBJECTS: Twenty healthy male volunteers participated in the study. DESIGN: The study was performed as a cross-over design. The subjects ingested corn peptide, and 30 min later alcohol was given. The blood ethanol and plasma free amino acids were analyzed at fixed times during a 2 h observation period after the alcohol intake. RESULTS: In subjects not given corn peptide, the blood ethanol concentration reached the maximum level of 9.35 mM at 30 min after alcohol intake, then gradually decreased. In addition, the plasma free amino acid levels decreased after alcohol intake. When subjects were given 3 and 5 g corn peptide 30 min prior to the alcohol intake, a significant reduction of the increase in blood ethanol levels of 3.7 mM and 2.83 mM was observed. In subjects given 5 g corn peptide, the plasma alanine, leucine, proline and free amino acid levels significantly elevated and remained at a high level during a 2 h observation period after alcohol intake. CONCLUSION: These results indicate that the ingestion of corn peptide reduces the increase in blood ethanol level after alcohol intake and causes the elevation in plasma amino acids alanine, leucine, and proline.

Administration, Oral↗
PubMed1996

[The analysis of peripheral blood lymphocytes of patients with silicosis and effects of silica in vitro].

It is well known that patients with silicosis are frequently associated with hyperglobulinemia, RA or PSS. It is also suggested that silica can drive and activated the immune system. In this study, we intended to measure the percentage of CD4+CD45RA+ cells in the peripheral blood of patients with silicosis and to investigate whether silica can actually activate human lymphocytes in vitro or not. Peripheral blood from 45 patients with silicosis was stained with OKT4 and 2H4 monoclonal antibodies, and measured by FACS analysis at 1st day and also checked similarly after 6 days-incubation without patients' sera. The intracellular Ca++ level was also checked after incubation with the normal human lymphocytes with silica by FACS analysis, using Fluo3-AM. CD4+CD45RA+ T cells decreased significantly in number. One to 5 minutes after the incubation with silica, intracellular Ca++ level increased markedly, which means the activation of lymphocytes in vitro. These results revealed that silica can activate human lymphocytes in vitro, but the role in vivo remains to be clarified.

Aged↗
PubMed1996

[89Sr Therapy for pain relief in patients with bone metastases].

Radioactive strontium chloride (89Sr) was administered for pain relief in 6 patients with bone metastases (4 prostate cancer and 2 breast cancer patients). Out of 6 patients, 2 showed apparent relief of bone pain and improvement of QOL, and 3 showed slight relief of the pain with or without improvement of QOL; that is, 83% was effective. Side effects were seen in 2 patients; transient deterioration of bone pain in one patient and bone marrow suppression in the other patient. The patient who showed bone marrow suppression had rather more lesions of bone metastasis (diffuse metastasis) and least urinary excretion of the radioactivity. Urinary excretion for 2 days varied 5 to 40% of the administered dose and was less in the patients with more metastatic lesions.

Bone Marrow↗
PubMed1996

Comparison of heavy metal concentrations in human umbilical cord in 1980 and 1990.

The well-known Minamata disease was caused by mercury; the Itai-Itai disease by cadmium; and lead poisoning by gasoline additives. Following our previous investigation on heavy metal concentrations in the umbilical cords in 1980, total mercury, cadmium and lead concentrations in the umbilical cords (from 20 males and 20 females) have been measured in 1990 for comparison in the present study. The changes in metal concentrations in 1980 and 1990 were: from 0.007 +/- 0.005 microgram/g to 0.011 +/- 0.008 microgram/g for total mercury; from 0.019 +/- 0.016 microgram/g to 0.006 +/- 0.005 microgram/g, from 0.151 +/- 0.123 microgram/g to 0.046 +/- 0.038 microgram/g for lead. Total mercury accumulated in the umbilical cords increased to approximately 1.6-fold, while cadmium and lead decreased to approximately 1/3-fold during these 10 years.

Adult↗
PubMed1995

Chemistry and antimicrobial activity of caryoynencins analogs.

Caryoynencins (1) are antibiotics isolated from liquid cultures of a plant pathogen, Pseudomonas caryophylli, and are unstable C18 carboxylic acids with a conjugated dienetetrayne structure. Enyne analogs of caryoynencins were synthesized from monosilylated 1,3-butadiyne 2 (n = 2), 1,3,5-hexatriyne 2 (n = 3), and 1,3,5,7-octatetrayne 2 (n = 4) by alkynyl metal addition to 2,4-hexadienal (3) followed by allylic rearrangement and deprotection. Tetraynol 5 (n = 4) thus obtained was resolved by enzyme reactions. The conjugated dienetetrayne compounds are mixtures of 3E,5E- and 3E,5Z-isomers, which equilibrate by room light. 13C-NMR chemical shifts of polyynes obey simple rules, which can be used for signal assignments. Antimicrobial activities of conjugated enynes and related compounds were examined. The tetrayne analog 6 (n = 4) possesses potent antibacterial and antifungal activities, while triyne and diyne analogs 6 (n = 3 and 2) are less active. Chirality does not affect the activities. An isomeric enyne compound, 2,4-tetradecadiene-7,9,11,13-tetrayn-6-ol (8), showed potent activity against Tricophyton.

Alkynes↗
PubMed1995

The DRE sequence TATCGATA, a putative promoter-activating element for Drosophila melanogaster cell-proliferation-related genes.

We have confirmed that the DNA replication-related element (DRE) consisting of an 8-bp palindrome, TATCGATA, and not neighboring sequences, are responsible for activating promoters of the Drosophila melanogaster (Dm) PCNA (proliferating cell nuclear antigen)- and DNA polymerase alpha-encoding genes in both cultured cell and transgenic fly systems. We have so far found 153 copies of DRE in the Dm gene database. 73 of them are concentrated within the 600-bp upstream regions from the transcription start points of 61 genes. Interestingly, many of these genes are involved in either DNA replication, transcription, translation, signal transduction, cell cycle or other putative regulatory functions, and are possibly related to cell proliferation. It seems likely that DRE is an element common to the regulation of cell-proliferation-related genes, although their expression patterns may be different depending on which of regulatory elements other than the DRE are combined.

Animals↗
PubMed1995

Frequent expression of P-glycoprotein/MDR1 by nasal T-cell lymphoma cells.

BACKGROUND: Lethal midline granuloma is now considered to be a malignant lymphoma derived from peripheral T cells or from natural killer cells. The therapeutic outcome of nasal T-cell lymphoma (NL) treated by conventional chemotherapy for non-Hodgkin's lymphoma is poor, although some patients have a good response to radiotherapy. To clarify the mechanisms of drug resistance, the expression of P-glycoprotein (P-gp)/MDR1, which is the product of the multidrug resistance (MDR) 1 gene, and MDR3 mRNA in NL cells, were examined. METHODS: Ten Japanese patients with NL were studied. Nine of these patients were examined before therapy. P-glycoprotein expression and phenotypes of lymphoma cells were examined by immunohistochemical staining using UIC2 as an anti-P-gp monoclonal antibody. In one case, the Rhodamine-123 efflux test was performed. MDR1 and MDR3 mRNA were detected by reverse transcription polymerase chain reaction. RESULTS: Nine of the 10 patients were P-gp positive. In one of nine, functional P-gp expression was observed. MDR1 mRNA was detected in all seven examined patients with P-gp positive NLs, whereas MDR3 mRNA was negative. Retrospectively, patients who received chemotherapy alone had poorer outcome than those treated by combination chemotherapy after irradiation. CONCLUSION: The poor prognosis for patients with NL treated with chemotherapy may be explained by P-gp expression of the NL cells.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed1995

Proton-translocating nicotinamide nucleotide transhydrogenase. Reconstitution of the extramembranous nucleotide-binding domains.

The nicotinamide nucleotide transhydrogenase of bovine mitochondria is a homodimer of monomer M(r) = 109,065. The monomer is composed of three domains, an NH2-terminal 430-residue-long hydrophilic domain I that binds NAD(H), a central 400-residue-long hydrophobic domain II that is largely membrane intercalated and carries the enzyme's proton channel, and a COOH-terminal 200-residue-long hydrophilic domain III that binds NADP(H). Domains I and III protrude into the mitochondrial matrix, where they presumably come together to form the enzyme's catalytic site. The two-subunit transhydrogenase of Escherichia coli and the three-subunit transhydrogenase of Rhodospirillum rubrum have each the same overall tridomain hydropathy profile as the bovine enzyme. Domain I of the R. rubrum enzyme (the alpha 1 subunit) is water soluble and easily removed from the chromatophore membranes. We have isolated domain I of the bovine transhydrogenase after controlled trypsinolysis of the purified enzyme and have expressed in E. coli and purified therefrom domain III of this enzyme. This paper shows that an active bidomain transhydrogenase lacking domain II can be reconstituted by the combination of purified bovine domains I plus III or R. rubrum domain I plus bovine domain III.

Animals↗
PubMed1995

Respiratory burst and tyrosine phosphorylation by vanadate.

We studied involvement of tyrosine-phosphorylated proteins in activation of NADPH oxidase in guinea pig neutrophils. Pervanadate, which is the oxidized form of orthovanadate, induced O2- production and protein tyrosine phosphorylation in neutrophils. O2- production induced by pervanadate was more sensitive to the tyrosine kinase-specific inhibitor, ST-638, as compared with the production induced by PMA. On the other hand, staurosporine more selectively inhibited PMA-induced O2- production than pervanadate-induced production. These results indicate that tyrosine kinase, not protein kinase C, is involved in pervanadate-induced O2- production. The tyrosine-phosphorylated proteins were detected in both the cytosol and membrane fractions prepared from pervanadate-induced neutrophils. In order to examine if tyrosine residues of some components of NADPH oxidase were directly phosphorylated, tyrosine-phosphorylated proteins were removed from solubilized membranes prepared from the pervanadate-stimulated neutrophils by immunoprecipitation with an anti-phosphotyrosine anti-body. NADPH oxidase activity in the solubilized membranes was not decreased by the treatment. These findings suggest that the components of NADPH oxidase are not tyrosine-phosphorylated by pervanadate treatment, that tyrosine phosphorylation may be involved in the signal transduction pathway of NADPH oxidase activation by pervanadate, and that this pathway is independent of the activation by protein kinase C.

Alkaloids↗
PubMed1995

Hyperphosphorylated p47-phox lost the ability to activate NADPH oxidase in guinea pig neutrophils.

p47-phox is one of the cytosolic activation factors of NADPH oxidase in neutrophils and known to translocate to plasma membranes and function by protein kinase C-phosphorylation. In cytosol fraction, prepared from calyculin A-treated neutrophils, the activity of cytosolic factor to activate NADPH oxidase was more reduced than that from PMA-treated cells. But, p47-phox did not translocate to the membranes, even if p47-phox was hyperphosphorylated in the calyculin A-treated neutrophils. Such hyperphosphorylated p47-phox seemed to lose the activity to constitute NADPH oxidase complex.

Animals↗
PubMed1995

Essential role of E2F recognition sites in regulation of the proliferating cell nuclear antigen gene promoter during Drosophila development.

We have found sequences similar to the transcription factor E2F recognition site within the Drosophila proliferating cell nuclear antigen (PCNA) gene promoter. These sequences are located at positions -43 to -36 (site I). and -56 to -49 (site II) with respect to the cap site Glutathione S-transferase (GST)-E2F and GST-DP fusion proteins cooperate and bind to the potential E2F sites in the PCNA promoter in vitro. A binding factor(s) to these sequences that has similar binding specificity to that of E2F was detected in nuclear extracts of Drosophila Kc cells. Furthermore, transient expression of target site for the activation coincided with the E2F sites. These results indicate that the PCNA gene is a likely target gene of E2F. Examination of lacZ expression from PCNA-lacZ fusion genes carrying mutations in either or both of two E2F sites introduced into flies by germ line transformation revealed that site II plays a major role in the PCNA promoter activity during embryogenesis and larval development, although both sites are required for optimal promoter activity. However, for maternal expression in ovaries, either one of the two sites is essentially sufficient to direct optimal promoter activity. These results demonstrate, for the first time, an essential role for E2F sites in regulation of PCNA promoter activity during development of a multicellular organism.

Animals↗
PubMed1995

Genomic cloning and chromosomal assignment of rat regucalcin gene.

The gene for a Ca(2+)-binding protein regucalcin was cloned from a rat genomic library which was constructed in lambda FIX II by screening with radiolabeled probe (complementary DNA of rat liver regucalcin). Positive clone had 19.9 kb insert of size and contained four exons of the gene coding for a rat regucalcin. These exons included the partial coding sequence (61.2% of open reading frame) and the entire 3'-untranslated region of the gene. The nucleotide sequence of exons completely agreed with that of a rat regucalcin cDNA clone. The sequence analysis of the clone showed that the identifier sequence and two simple repeated sequences exist in the intron of the gene. Moreover, chromosomal location of the rat regucalcin gene was determined by direct R-banding fluorescence in situ hybridization (FISH) method with the 19.9 kb clone containing four exons. The regucalcin gene was localized on rat chromosome Xq11.1-12 proximal end.

Amino Acid Sequence↗
PubMed1995

Increase in cystine transport activity and glutathione level in mouse peritoneal macrophages exposed to oxidized low-density lipoprotein.

The transport of cystine has been investigated in mouse peritoneal macrophages incubated with oxidized low-density lipoprotein (oxi-LDL; low-density lipoprotein, LDL). The transport activity for cystine was potently induced by oxi-LDL but not by native or acetylated LDL. The response of the cells to oxi-LDL was dependent on the extent of oxidative modification of LDL. The transport activity for other amino acids was not induced by oxi-LDL. GSH content increased in macrophages incubated with oxi-LDL and this increase was accounted for by the induction of the cystine transport activity because the increase was completely blocked by glutamate or homocysteate which shared the transport system for cystine and thus inhibited the uptake of cystine competitively.

Animals↗
PubMed1995

Stimulatory effect of hormonal signaling factors on (Ca(2+)-Mg2+)-ATPase activity in rat liver plasma membranes: cross talk with regucalcin.

The effect of hormonal signaling factors on (Ca(2+)-Mg2+)-ATPase activity in rat liver plasma membranes was investigated. The presence of inositol-glycan (10(-7)-10(-5) M), dibutyryl cAMP (10(-4) and 10(-3) M) or inositol 1,4,5-trisphosphate (IP3; 10(-6) and 10(-5) M) in the enzyme reaction mixture produced a significant increase in (Ca(2+)-Mg2+)-ATPase activity. These effects were completely inhibited by the presence of vanadate (10(-4) M), an inhibitor of the enzyme phosphorylation, and N-ethylmaleimide (5 x 10(-3) M), a SH group modifying reagent. Meanwhile, regucalcin, a Ca(2+)-binding protein isolated from rat liver cytosol, increased the enzyme activity by binding to the SH groups of (Ca(2+)-Mg2+)-ATPase in liver plasma membranes. The presence of regucalcin (0.25 microM) with an effective concentration completely inhibited the effect of inositol-glycan (10(-5) M) to increase (Ca(2+)-Mg2+)-ATPase activity, while the effect of dibutyryl cAMP (10(-3) M) or IP3 (10(-5) M) was not altered. The inositol-glycan effect was not modulated by the presence of dibutyryl cAMP or IP3. Now, the preincubation of the plasma membranes with regucalcin did not modify the effect of inositol-glycan on the enzyme activity, suggesting that regucalcin competes with inositol-glycan for the binding to the plasma membranes. The present results suggest that there may be a cross talk with regucalcin and hormonal signaling factors in the regulation of (Ca(2+)-Mg2+)-ATPase activity in liver plasma membranes.

Animals↗