Isoform-specific effects on the interaction of apolipoprotein E with -amyloid: implication of the mechanism by which apolpipoprotein E contributes to development of Alzheimerz’s disease. 2001(Mar.) Author:山内, 一由
Automation and New Technology in the Clinical Chemistry 1990(Oct.) Author:Shozo Nomoto; Kazuyoshi Yamauchi; Moritoshi Sato; F. William; Sunderman, Jr
論文 Cut-off values of serum IgG4 among three reagents, including a novel IgG4 reagent: a multicenter study Scientific Reports,11(1) 2021(Dec.) Author:Yoko Usami; Mitsutoshi Sugano; Takeshi Uehara; Masayoshi Koinuma; Nau Ishimine; Kenji Kawasaki; Kazuyoshi Yamauchi; Hideaki Hamano; Takayuki Honda Abstract:Abstract
Elevated serum IgG4 is a useful marker of IgG4-related disease (IgG4-RD) activity. However, there is no uniformity in the cut-off values of IgG4 among the various reagents. The aim of this study was to compare the measured and cut-off values of IgG4 assessed using three different reagents. This study enrolled 466 IgG4-RD and non-IgG4-RD patients who required measurement of serum IgG4 levels to diagnose or treat IgG4-RD. Serum IgG4 was measured using three reagents: N-assay LA IgG4 Nittobo (Nittobo), BS-NIA IgG4 (TBS), and N Latex IgG4 (Siemens). The values obtained using the three reagents were compared, and cut-off values were calculated for each. Although there was good correlation among the results with the three reagents, the measured and cut-off values were all different. The Nittobo values were 1.4 times the TBS values and the TBS values were almost half those of the Siemens values. ROC curve analysis showed cut-off values for the Nittobo, TBS, and Siemens reagents of 1.42, 1.31, and 2.38 g/L, respectively. The measured and cut-off values of serum IgG4 vary depending on the reagents used for the assay, although there is good correlation among the values measured by the three reagents.
A novel variant fibrinogen, AαE11del, demonstrating the importance of AαE11 residue in thrombin binding International Journal of Hematology,114(5):591-598 2021(Nov.) Author:Takahiro Kaido; Masahiro Yoda; Tomu Kamijo; Shinpei Arai; Kazuyoshi Yamauchi; Nobuo Okumura
Redox index of Cys-thiol residues of serum apolipoprotein E and its diagnostic potential Bioscience Reports,41(8) 2021(Aug. 27) Author:Kazuyoshi Yamauchi; Chiaki Taira; Yasushi Kawakami Abstract:Abstract
Background: The redox modulation of Cys-thiol participates in various pathophysiological processes. We explored the proper index for estimating the redox status of Cys-thiol of serum apolipoprotein E (apoE), named “redox-IDX-apoE,” which is necessary to understand the redox biology of age-related diseases.
Methods: The fractions of the reduced form (red-), reversible oxidized form (roxi-), and irreversibly oxidized form (oxi-) apoE in serum, obtained from the patients with no apparent disease (controls, n=192) and with atherosclerosis and type 2 diabetes (patients, n=16), were measured by a band-shift assay using a maleimide compound. Redox-IDX-apoE candidates were determined by calculating the values of these fractions and the total apoE concentration.
Results: Cys number of apoE significantly increased for the ratio of roxi-apoE to total-apoE (roxi/total) (E2/E3>E3/E3>E3/E4) but decreased for the ratios of red-apoE to roxi-apoE (red/roxi) and [red-apoE + oxi-apoE] to roxi-apoE ([red + oxi]/roxi) (E2/E3<E3/E3<E3/E4). Considering the subjects with apoE3/E3, these ratios were independent of age and sex. Roxi/total showed negative correlations with serum triglyceride (TG) and HbA1c levels, while both red/roxi and [red + oxi]/roxi showed significant positive correlations with them. However, red/roxi and [red + oxi]/roxi in patients were significantly lower than those in controls, although serum TG and HbA1c levels in the patients were significantly higher than those in controls.
Conclusion: The redox status of serum apoE-Cys-thiol is closely involved in the metabolism of TG-rich lipoproteins and glucose. The appropriate use of redox-IDX-apoE could be helpful in the diagnosis and prognosis of age-related diseases and in understanding the underlying mechanisms.
Recombinant γY278H Fibrinogen Showed Normal Secretion from CHO Cells, but a Corresponding Heterozygous Patient Showed Hypofibrinogenemia International Journal of Molecular Sciences,22(10):5218-5218 2021(May 14) Author:Tomu Kamijo; Takahiro Kaido; Masahiro Yoda; Shinpei Arai; Kazuyoshi Yamauchi; Nobuo Okumura Abstract:We identified a novel heterozygous hypofibrinogenemia, γY278H (Hiroshima). To demonstrate the cause of reduced plasma fibrinogen levels (functional level: 1.12 g/L and antigenic level: 1.16 g/L), we established γY278H fibrinogen-producing Chinese hamster ovary (CHO) cells. An enzyme-linked immunosorbent assay demonstrated that synthesis of γY278H fibrinogen inside CHO cells and secretion into the culture media were not reduced. Then, we established an additional five variant fibrinogen-producing CHO cell lines (γL276P, γT277P, γT277R, γA279D, and γY280C) and conducted further investigations. We have already established 33 γ-module variant fibrinogen-producing CHO cell lines, including 6 cell lines in this study, but only the γY278H and γT277R cell lines showed disagreement, namely, recombinant fibrinogen production was not reduced but the patients’ plasma fibrinogen level was reduced. Finally, we performed fibrinogen degradation assays and demonstrated that the γY278H and γT277R fibrinogens were easily cleaved by plasmin whereas their polymerization in the presence of Ca2+ and “D:D” interaction was normal. In conclusion, our investigation suggested that patient γY278H showed hypofibrinogenemia because γY278H fibrinogen was secreted normally from the patient’s hepatocytes but then underwent accelerated degradation by plasmin in the circulation.
Balance capability and smell identification are associated with cognitive function in middle-aged persons with type 2 diabetes DIABETOLOGIA,63(SUPPL 1::1:SI):S455-S456 2020(Sep.) Author:Suzuki, H; Midorikawa, M; Suzuki, Y; Sato, H; Nemoto, K; Yamauchi, K; Sugano, Y; Osaki, Y; Iwasaki, H; Sekiya, M; Yahagi, N; Hada, Y; Arai, T; Shimano, H
The redox status of cysteine thiol residues of apolipoprotein E impacts on its lipid interactions Biological chemistry,401(5):617-627 2020(Feb.) Author:Yamauchi, Kazuyoshi; Kawakami, Yasushi Abstract:Redox-mediated modulation of cysteine (Cys) thiols has roles in various pathophysiological functions. We recently found that formation of disulfide-linked complexes of apolipoprotein (apo) E3 prevented apoE3 from irreversible oxidation. In this report, the influence of modification of Cys thiols in apoE2 and apoE3 on interactions with lipids was investigated. The apoE redox status was examined by a band-shift assay using a maleimide compound, and interactions with lipids were evaluated by a kinetic assay using dimyristoyl-snglycero-3-phosphocholine (DMPC) and non-denaturing polyacrylamide gel electrophoresis. A reduction in DMPC clearance activity of apoE2 and apoE3 but not apoE4 was observed. Although hydrogen peroxide-induced oxidation decreased the clearance activity of the isoforms, apoE2 showed the greatest residual activity. Both Cys thiol masking and dimerization decreased the activity of apoE2 and apoE3 but not apoE4. In contrast, apoAII preincubation markedly increased the activity (apoE2 > apoE3 > apoE4), in accordance with the formation of apoE-AII and apoAII-E2-AII complexes. ApoAII preincubation also reduced the particle size of apoEDMPC liposome complexes, especially
Evaluation of a novel serum IgG4 assay and determination of reference interval for the Japanese population Clinica chimica acta; international journal of clinical chemistry,501:136-141 2019(Nov.) Author:Usami, Yoko; Ichihara, Kiyoshi; Uehara, Takeshi; Sugano, Mitsutoshi; Ishimine, Nau; Kawasaki, Kenji; Yamauchi, Kazuyoshi; Hamano, Hideaki; Honda, Takayuki Abstract:[BACKGROUND] IgG4-related disease (IgG4-RD) is a new syndrome characterized by elevated serum IgG4 concentration and tissue infiltration of IgG4-positive plasma cells. Here, we evaluated the analytical performance of a new IgG4 assay reagent featuring a wide dynamic range, highly specific monoclonal antibody, and the reversed passive latex agglutination assay and determined the IgG4 reference interval (RI) for the Japanese population.
[METHODS] Performance evaluations were conducted on precision, linearity, sensitivity, interference, and method comparison with The Binding Site (TBS) and Siemens reagents. The RI was derived by the parametric method from 619 apparently healthy Japanese 18 to 65 years of age.
[RESULTS] Between-day precisions ranged from 1.99 to 5.52 CV%. Linearity was confirmed up to 5.0 g/l. The limit of quantitation was 0.085 g/l. Interfering substances did not significantly influence values. Method comparison among the 3 reagents yielded correlation coefficients between 0.973 and 0.988. Values for the new reagent matched those of TBS reagent except at a higher concentration range, where reactivity dissociated. The RI was 0.11-1.21 g/l without distinction by s
Redox equilibrium of serum apolipoprotein E3: A buffering effect of disulfide-linked complexes against oxidative stress on apolipoprotein E3-containing lipoproteins Bioscience reports,39(4) 2019(Apr.) Author:Yamauchi, Kazuyoshi; Iwasaki, Shio; Kawakami, Yasushi Abstract:Reversible redox modification of cysteine thiols is crucial for protecting proteins from irreversible detrimental change. However, the physiological significance of the redox modification of apolipoprotein (apo) E is unclear. Here, we hypothesized that the disulfide-linked complexes of apoE3 corresponding to the representative reversible-modified apoE3 play a protective role against oxidative stress. The effects of disulfide bond formation on oxidative stress on apoE3 were evaluated with a band-shift assay. Maleimide-labeled apoE3 and unlabeled apoE3 were defined as the reduced (r)-apoE3 and non-reduced (nr)-apoE3 forms, respectively. Hydrogen peroxide-induced oxidation decreased for r-apoE3 but increased for nr-apoE3. Induction of apoE3-AII complex formation with excess of apoAII markedly suppressed the oxidative stress-induced increase in nr-apoE3 ( <0.001) and enhanced homodimer formation. The apoE3-AII complex was more dominant in high-density lipoprotein (HDL) than in very low-density lipoprotein. Under oxidative stress, HDL showed a significant decrease, rather than an increase, in nr-apoE3 levels with a concomitant significant increase in apoE3-AII levels ( <0.005). This f
Redox status of serum apolipoprotein E and its impact on HDL cholesterol levels Clinical Biochemistry,50(13-14):777-783 2017(Sep. 01) Author:Kazuyoshi Yamauchi; Yuka Ebihara; Yasushi Kawakami Abstract:Background Apolipoprotein E (apoE) is closely involved in the pathogenesis of apoE-related diseases, such as Alzheimer's disease and cardiovascular disease. The redox modulation of cysteine-thiols in a protein is involved in various pathophysiological regulations
however, that of apoE has not been studied in detail. Herein, we devised an analytical method to determine the redox status of serum apoE and assessed its relation to serum cholesterol levels and apoE phenotype. Methods The present method was based on a band shift assay, using a photocleavable maleimide-conjugated polyethylene glycol. Results The basic characteristics of the present method were found to be satisfactory to determine the redox status of serum apoE quantitatively. Serum apoE was separated into its reduced-form (r-), non-reduced-form (nr-), apoE-AII complex, and homodimer using this method. R-apoE could be detected as a 40-kDa band, whereas nr-apoE remained as monomeric apoE. R-apoE displayed a preference for VLDL
however, the levels showed the correlation with HDL-cholesterol levels (p <
0.005). Redox status of serum apoE was significantly different among apoE phenotypes. The quantitative ratios of nr-apoE to total apoE in serum from subjects with apoE4/E3 were higher than in serum from subjects with apoE3/E3 (p <
0.0001) and apoE3/E2 (p <
0.001). Conclusion The redox status of serum apoE might be related to the synthesis of HDL. The information concerning the redox status of serum apoE provided by the present method may be a potent indicator to evaluate various apoE-related diseases.
Impact of TLR 2, TLR 4-activation on the Expression of ABCA1 and ABCG1 in Raw Cells ANNALS OF CLINICAL AND LABORATORY SCIENCE,47(4):436-446 2017(Jul.) Author:Kana Suzuki; Yasushi Kawakami; Kazuyoshi Yamauchi Abstract:Toll-like receptors (TLR) activation is thought to modulate the macrophage cholesterol efflux and contribute to the atherosclerosis progression; however, the precise pathophysiological mechanism remains unclear. We investigated the effects of TLR2-and TLR4-activation on the expression of the ATP-binding cassette (ABC) transporters ABCA1 and ABCG1 in a mouse macrophage cell line, Raw 264.7. Both TLR2-and TLR4-activation upregulated the expression of ABCA1 mRNA but downregulated that of ABCG1 mRNA. These alterations may be mainly regulated by the following 3 cascades: (1) the TLR/myeloid differentiation primary-response protein 88/Liver X receptor pathway, which upregulated the ABCA1 mRNA; (2) NF-kappa B pathway, which downregulated the ABCG1 mRNA, and (3) the p38 pathway, which upregulated and stabilized ABCA1 mRNA. These cascades are involved in a complex crosstalk and result in the upregulation of ABCA1 mRNA without a change in ABCA1 protein and the down-regulation of ABCG1 mRNA leading to the increase in ABCG1 protein. These alterations, especially the induction of ABCG1 protein, may be closely involved with the development of atherosclerosis.
The Asian project for collaborative derivation of reference intervals: (2) results of non-standardized analytes and transference of reference intervals to the participating laboratories on the basis of cross-comparison of test results CLINICAL CHEMISTRY AND LABORATORY MEDICINE,51(7):1443-1457 2013(Jul.) Author:Kiyoshi Ichihara; Ferruccio Ceriotti; Mori Kazuo; Yang-Yang Huang; Yoshihisa Shimizu; Haruki Suzuki; Masami Kitagawa; Kazuyoshi Yamauchi; Sadao Hayashi; Chia-Chun Tsou; Yoshikazu Yamamoto; Shigeo Ishida; Linda Leong; Michitaka Sano; Hwan Sub Lim; Akira Suwabe; Hee-Yeon Woo; Keiya Kojima; Yoshio Okubo Abstract:Background: The 2009 Asian multicenter study for derivation of reference intervals (RIs) featured: 1) centralized measurements to exclude reagent-dependent variations; 2) inclusion of non-standardized analytes (hormones, tumor makers, etc.) in the target; and 3) cross-check of test results between the central and local laboratories. Transferability of centrally derived RIs for non-standardized analytes based on the cross-check was examined.
Methods: Forty non-standardized analytes were centrally measured in sera from 3541 reference individuals recruited by 63 laboratories. Forty-four laboratories collaborated in the cross-check study by locally measuring aliquots of sera from 9 to 73 volunteers (average 22.2). Linear relationships were obtained by the reduced major-axis regression. Error in converting RIs using the regression line was expressed by the coefficient of variation of slope b[CV(b)]. CV(b) <10% was set as the cut-off value allowing the conversion. The significance of factors for partitioning RIs was determined similarly as in the first report.
Results: Significant sex-, age-, and region-related changes in test results were observed in 17, 15, and 11 of the 40 analytes, respectively. In the cross-comparison study, test results were not harmonized in the majority of immunologically measured analytes, but their average CV(b)s were <10% except for total protein, cystatin C, CA19-9, free thyroxine, and triiodothyronine. After conversion, 74% of centrally derived RIs were transferred to each local laboratory.
Conclusions: Our results point to the feasibility of: 1) harmonizing test results across different laboratories; and 2) sharing centrally derived RIs of non-standardized analytes by means of comparative measurement of a set of commutable specimens.
Waldenström macroglobulinemia transforming from t (11;18) (q21;q21) -negative gastric MALT lymphoma after systemic dissemination. Rinsho Byori 2012(Jun.) Author:Yamauchi,Kazuyoshi
Characterization of CIA-1, an Ambler Class A Extended-Spectrum beta-Lactamase from Chryseobacterium indologenes ANTIMICROBIAL AGENTS AND CHEMOTHERAPY,56(1):588-590 2012(Jan.) Author:Takehisa Matsumoto; Mika Nagata; Nau Ishimine; Kenji Kawasaki; Kazuyoshi Yamauchi; Eiko Hidaka; Eriko Kasuga; Kazuki Horiuchi; Kozue Oana; Yoshiyuki Kawakami; Takayuki Honda Abstract:An Ambler class A beta-lactamase gene, bla(CIA-1), was cloned from the reference strain Chryseobacterium indologenes ATCC 29897 and expressed in Escherichia coli BL21. The bla(CIA-1) gene encodes a novel extended-spectrum beta-lactamase (ESBL) that shared 68% and 60% identities with the CGA-1 and CME-1 beta-lactamases, respectively. bla(CIA-1)-like genes were detected from clinical isolates. In addition to the metallo-beta-lactamase IND of Ambler class B, C. indologenes has a class A ESBL gene, bla(CIA-1), located on the chromosome.
Pathophysiological Investigation of the Gastric Surface Mucous Gel Layer of Patients with Helicobacter pylori Infection by Using Immunoassays for Trefoil Factor Family 2 and Gastric Gland Mucous Cell-Type Mucin in Gastric Juice DIGESTIVE DISEASES AND SCIENCES,56(12):3498-3506 2011(Dec.) Author:Seiko Kubota; Kazuyoshi Yamauchi; Mitsutoshi Sugano; Kenji Kawasaki; Atsushi Sugiyama; Kenji Matsuzawa; Taiji Akamatsu; Yasukazu Ohmoto; Hiroyoshi Ota Abstract:Background The trefoil factor family (TFF) 2 protein is produced by gastric gland mucous cells (GMCs), and the secreted TFF2 shares a mucosal barrier function with GMC-type mucin. Recently, we presented an enzyme-linked immunosorbent assay (ELISA) method for measurement of GMC-type mucin in the gastric juice.
Aims We aimed to develop an ELISA for TFF2 and to assess pathophysiological changes in the gastric surface mucous gel layer (SMGL) of patients with Helicobacter pylori infection.
Methods The distribution of TFF2 and GMC-type mucin in the SMGL was immunohistochemically determined. The ELISA for TFF2 was based on a polyclonal goat antibody. Recombinant TFF2 was employed to prepare the calibrators. TFF2 and GMC-type mucin in the gastric juice in healthy individuals (n = 33) and patients with gastritis (n = 37), gastric ulcer (n = 16), and duodenal ulcer (n = 10) were assayed using ELISA.
Results TFF2 and GMC-type mucin were immunohistochemically co-localized in the gastric SMGL and GMCs. The TFF2 levels in the patients were significantly higher than those in the healthy individuals. Further, the TFF2 levels in the H. pylori-positive patients were significantly higher than those in the H. pylori-negative patients, and decreased after the eradication of the infection. GMC-type mucin levels showed a tendency similar to that of TFF2 levels.
Conclusions The upregulation of TFF2 and GMC-type mucin secretion may reflect the response of the gastric mucosa to H. pylori-induced injuries. TFF2 and GMC-type mucin secreted into the SMGL may protect the gastric mucosa against H. pylori.
Factor H gene variants in Japanese: Its relation to atypical hemolytic uremic syndrome MOLECULAR IMMUNOLOGY,49(1-2):48-55 2011(Oct.) Author:Saki Mukai; Yoshihiko Hidaka; Masako Hirota-Kawadobora; Kazuyuki Matsuda; Noriko Fujihara; Yuka Takezawa; Seiko Kubota; Kenichi Koike; Takayuki Honda; Kazuyoshi Yamauchi Abstract:Mutations and polymorphisms of factor H gene (FH1)are known to be closely involved in the development of atypical hemolytic uremic syndrome (aHUS). Several groups have identified disease risk mutations and polymorphisms of FH1 for the development of aHUS, and have investigated frequencies of aHUS in a number of ethnic groups. However, such studies on Japanese populations are limited. In the present study, we analyzed FH1 in Japanese aHUS patients and healthy volunteers, and examined whether those variants impacted on a tendency for the development of aHUS in Japanese populations. Similar to previous studies, we found that a high frequency of FH1 mutations, located in exon 23 of FH1, encodes short consensus repeat 20 in C-terminal end of factor H molecule in patients with aHUS (40%), but not in healthy volunteers. Interestingly, no significant differences in frequency of well-known disease risk polymorphisms for aHUS were observed between healthy volunteers and aHUS patients. Our results suggested that although FH1 mutations relates to the development of Japanese aHUS in accordance with other ethnic studies, other factor may be required for factor H polymorphism to be a risk factor of Japanese aHUS. (C) 2011 Elsevier Ltd. All rights reserved.
Detection of chymase-digested C-terminally truncated apolipoprotein A-I in normal human serum JOURNAL OF IMMUNOLOGICAL METHODS,369(1-2):51-58 2011(Jun.) Author:Yoko Usami; Kazuyuki Matsuda; Mitsutoshi Sugano; Nau Ishimine; Yuriko Kurihara; Tamaki Sumida; Kazuyoshi Yamauchi; Minoru Tozuka Abstract:In atherosclerotic artery walls, mast cells, an inflammatory cell, are activated and secrete some proteases including chymase. Chymase, a chymotrypsin-like protease, cleaves the C-terminus of apolipoprotein A-I (apoA-I) at Phe225. This cleavage reduces the ability of apoA-I to promote the efflux of cellular cholesterol. The aim of this study is to detect C-terminally truncated apoA-I in normal human serum. For this purpose, we generated a monoclonal antibody that specifically recognizes C-terminally truncated apoA-I by immunizing mice with a peptide that corresponds to human apoA-I amino acid residues 216-225. The monoclonal antibody, termed 16-4 mAb, selectively reacted with recombinant C-terminally truncated apoA-I, but not recombinant full-length apoA-I. A two-dimensional electrophoresis analysis also indicated that only two out of six spots that contained apoA-I fragments and had a molecular mass of 26 kDa after chymase digestion reacted with the 16-4 mAb. We detected an extremely small amount of C-terminally truncated apoA-I in normal human serum by concentrating the serum through affinity chromatography using a 16-4 mAb-conjugated resin, and then performing Western blot analysis. The 16-4 mAb could be useful to examine whether C-terminally truncated apoA-I is associated with the progression of atherosclerosis. (C) 2011 Elsevier B.V. All rights reserved.
Quantitative monitoring of single nucleotide mutations by allele-specific quantitative PCR can be used for the assessment of minimal residual disease in patients with hematological malignancies throughout their clinical course CLINICA CHIMICA ACTA,412(1-2):53-58 2011(Jan.) Author:Chiaki Taira; Kazuyuki Matsuda; Yuka Kamijyo; Kazuo Sakashita; Fumihiro Ishida; Toshiko Kumagai; Kazuyoshi Yamauchi; Nobuo Okumura; Takayuki Honda Abstract:Background: Monitoring of minimal residual disease (MRD) in patients with hematological malignancies is important for evaluating the patients' therapeutic response and risk of relapse. Single nucleotide mutations associated with leukemogenesis can be considered as applicable MRD markers.
Methods: We developed an allele-specific quantitative polymerase chain reaction (AS-qPCR) for FLT3 2503G>T, KIT2446G>T, and KIT2447A>T and compared the change in the expression levels of the FLT3 or KIT mutations assessed by AS-qPCR to those of the RUNX1-RUNX1T1 fusion gene and WT1 by conventional quantitative PCR.
Results: The AS-qPCR using primers including template-mismatched nucleotide or template-mismatched nucleotide plus locked nucleic acid substituted nucleotide provided higher selectivity for mutant nucleotides. The change in the expression levels of the FLT3 or KIT mutations at the time of relapse and just after hematopoietic stem cell transplantation correlated well with that of the RUNX1-RUNX1T1 fusion gene and WT1. Moreover, during complete remission, only AS-qPCR could detect low-level expression of residual mutations.
Conclusions: The AS-qPCR for analyzing single nucleotide mutations contributes to the monitoring of MRD in patients without recurrent fusion gene throughout the clinical course and thus broadens the spectrum of patients in whom MRD can be monitored. (C) 2010 Elsevier B.V. All rights reserved.
In vitro transcription of compound heterozygous hypofibrinogenemia Matsumoto IX; first identification of FGB IVS6 deletion of 4 nucleotides and FGG IVS3-2A > G causing abnormal RNA splicing CLINICA CHIMICA ACTA,411(17-18):1325-1329 2010(Sep.) Author:Fumiko Terasawa; Yuka Kamijyo; Noriko Fujihara; Kazuyoshi Yamauchi; Toshiko Kumagai; Takayuki Honda; Satoshi Shigematsu; Nobuo Okumura Abstract:Background: We reported a case of hypofibrinogenemia Matsumoto IX (M IX) caused by a novel compound heterozygous mutation involving an FGB IVS6 deletion of 4 nucleotides (Delta 4b) (three T, one G; between FGB IVS6-10 and -16) and FGG IVS3-2A/G, which are both identified for the first time. To examine the transcription of mRNA from the M IX gene, we cloned the wild-type and mutant genes into expression vectors.
Methods: The vectors were transfected into CHO cells and transiently produced wild-type, B beta-, or gamma-mRNA in the cells. The mRNAs amplified with RT-PCR were analyzed by agarose gel electrophoresis and nucleotide sequencing.
Results: The RT-PCR product from FGB IVS6 Delta 4b showed aberrant mRNA that included both introns 6 and 7, and that from FGG IVS3-2G showed two aberrant mRNAs, a major one including intron 3 and a minor in which intron 3 was spliced by a cryptic splice site in exon 4. We speculated that the aberrant mRNAs are degraded before translation into proteins, and/or translated variant chains are subjected to quality control and degraded in the cytoplasm.
Conclusion: The reduced plasma fibrinogen level of the M IX patient was caused by abnormal RNA splicing of one or both of the FGB and FGG genes. (C) Elsevier B.V. All rights reserved.
Identification of N-homocysteinylated apolipoprotein Al in normal human serum ANNALS OF CLINICAL BIOCHEMISTRY,47(Part 5):453-459 2010(Sep.) Author:N. Ishimine; Y. Usami; S. Nogi; T. Sumida; Y. Kurihara; K. Matsuda; K. Nakamura; K. Yamauchi; N. Okumura; M. Tozuka Abstract:Background: In human serum, a portion of homocysteine (Hcy) exists as an N-linked form to the epsilon-amino group of protein lysine residues. N-homocysteinylated proteins differ structurally and functionally from native proteins. The present study strives to develop detection and potential semi-quantification methods for N-homocysteinylated apolipoprotein Al (N-HcyapoAl) in human serum.
Methods: Serum treated with or without cysteamine was supplied to isoelectric focusing (IEF) followed by an immunoblot using an anti-apoAl antibody. Cysteamine treatment increased the isoelectric point for N-Hcy-apoAl, but not for unmodified apoAl, due to the presence of -SH group(s) derived from Hcy and the absence of a cysteine residue in the apoAl molecule. N-Hcy-apoAl was semi-quantified from the scanned immunoblot pattern via a computer.
Results: After cysteamine treatment, N-Hcy-apoAl in the serum was identified by IEF at the position with a higher pl value compared with intact apoAl. The reproducibility (between assays) of the semi-quantification method was 19.1 % CV (coefficient of variation) for an average ratio 5.9% of N-Hcy-apoAl to the whole apoAl in the serum. Approximately 1.0-7.4% of apoAl was N-homocysteinylated in the serum obtained from 27 healthy subjects. Neither the ratio of N-Hcy-apoAl nor its concentration, calculated by total apoAl concentration, indicated correlation with the so-called total (free and S-linked) Hcy concentration.
Conclusions: We directly found that a portion of apoAl in the serum undergoes homocysteinylation in an N-linkage manner, and used this to develop a potential semi-quantification method for N-Hcy-apoAl.
A C-terminal amino acid substitution in the gamma-chain caused by a novel heterozygous frameshift mutation (Fibrinogen Matsumoto VII) results in hypofibrinogenaemia THROMBOSIS AND HAEMOSTASIS,104(2):213-223 2010(Aug.) Author:Noriko Fujihara; Ayumi Haneishi; Kazuyoshi Yamauchi; Fumiko Terasawa; Toshiro Ito; Fumihiro Ishida; Nobuo Okumura Abstract:We found a novel hypofibrinogenemia designated as Matsumoto VII (M-VII), which is caused by a heterozygous nucleotide deletion at position g.7651 in FGG and a subsequent frameshift mutation in codon 387 of the gamma-chain. This frameshift results in 25 amino acid substitutions, late termination of translation with elongation by 15 amino acids, and the introduction of a canonical glycosylation site. Western blot analysis of the patient's plasma fibrinogen visualised with anti-gamma-chain antibody revealed the presence of two extra bands. To identify the extra bands and determine which of the above-mentioned alterations caused the assembly and/or secretion defects in the patient, 11 variant vectors that introduced mutations into the cDNA of the gamma-chain or gamma'-chain were transfected into Chinese hamster ovary cells. In vitro expression of transfectants containing gamma Delta 7651A and gamma Delta 7651A/399T (gamma Delta 7651A with an amino acid substitution of 399Asn by Thr and a variant lacking the canonical glycosylation site) demonstrated a reduction in secretion to approximately 20% of the level seen in the transfectants carrying the normal gamma-chain. Furthermore, results from other transfectants demonstrated that eight aberrant residues between 391 and 398 of the M-VII variant, rather than the 15 amino acid extension or the additional glycosylation, are responsible for the reduced levels of assembly and secretion of M-VII variant fibrinogen. Finally, the results of this study and our previous reports demonstrate that the fibrinogen gamma-chain C-terminal tail (388-411) is not necessary for protein assembly or secretion, but the aberrant amino acid sequence observed in the M-VII variant (especially 391-398) disturbs these functions.
Analysis of plasmin generation and clot lysis of plasma fibrinogen purified from a heterozygous dysfibrinogenemia, bGly15Cys (Hamamatsu II). Blood Coagul Fibrinolysis,20(8):726-732 2009(Dec.) Author:Kamijyo, Y; Hirota-Kawadobora, M; Yamauchi, K; Terasawa, F; Honda, T; Ikeya, M; Okumura, N
Crucial roles of Helicobacter pylori infection in the pathogenesis of gastric cancer and gastric mucosa-associated lymphoid tissue (MALT) lymphoma Rinsho Byori,57(9):861-869 2009(Sep.) Author:Ota, H; Asano, N; Yamauchi, K; Akamatsu, T
Functional analysis of heterozygous plasma dysfibrinogens derived from two families of gammaArg275Cys and three families of gammaArg275His, and haplotype analysis for these families Rinsho Byori,57(7):651-658 2009(Jul.) Author:Kamijyo, Y; Hirota-Kawadobora, M; Fujihara, N; Wakabayashi, S; Matsuda, K; Yamauchi, K; Terasawa, F; Okumura, N; Honda, T
Recombinant variant fibrinogens substituted at residues gnmma326Cys and ganmma339Cys demonstrated markedly impaired secretion of assembled fibrinogen. Thromb Res 2009(May) Author:Haneishi, A; Terasawa, F; Fujihara, N; Yamauchi, K; Okumura, N; Katsuyama, T
Characterization of sialo-apolipoprotein E in human cerebrospinal fluid Rinsho Byori,57(5):436-444 2009(May) Author:Yamauchi, K
Laboratory appraisal of optimal gaseous conditions for growth of zoonotic Helicobacter felis ATCC 49179. Microbiology and Immunology,53(5):251-258 2009(May) Author:Shiohara, M; Kawakubo, M; Matsumoto, T; Kumagai, T; Yamauchi, K; Oana, K; Ota, H; Kawakami, Y
Characterization of cysteine and homocysteine bound to human serum transthyretin. Clin Chim Acta,402(1-2):61-66 2009(Apr.) Author:Hanyu, N; Shimizu, T; Yamauchi, K; Okumura, N; Hidaka, H
Sialic Acid Moiety of Apolipoprotein E and Its Impact on the Formation of Lipoprotein Particles in Human Cerebrospinal Fluid. Clinica Chimica Acta,402(1-2):61-66 2009(Apr.) Author:Kawasaki, K; Ogiwara, N; Sugano, M; Okumura, N; Yamauchi, K
Cryptic insertion into 11q23 of MLLT10 not involved in t(1;15;11;10)(p36;q11;q23;q24) in infant acute biphenotypic leukemia. Cancer Genet Cytogenet,15(190(2)):113-120 2009(Apr.) Author:Matsuda, K; Tanaka, M; Araki, S; Yanagisawa, R; Yamauchi, K; Koike, K
Phylogeny of a novel "Helicobacter heilmannii" organism from a Japanese patient with chronic gastritis based on DNA sequence analysis of 16S rRNA and urease genes. J Microbiol,47(2):201-207 2009(Apr.) Author:Matsumoto, T; Kawakubo, M; Shiohara, M; Kumagai, T; HIdaka, E; Yamauchi, K; Oana, K; Matsuzawa, K; Ota, H; Kawakami, Y
Simple allele-specific oligonucleotide polymerase chain reaction for the detection of mutations and deletions in the epidermal growth factor receptor gene: Applications of this method for the diagnosis of non-small-cell lung cancer. Clinica Chimica Acta,401:68-72 2009(Mar.) Author:Uhara, M; Matsuda, K; Taira, C; Higuchi, Y; Okumura, N; Yamauchi, K
Clinical Assessment of Novel ChromID ESBL Agar Plates for Detection of ESBL Producers in the Family Enterobacteriaceae. Rinsho Biseibutshu Jinsoku Shindan Kenkyukai Shi,20(1):1-9 2009(Jan.) Author:Kasuga, E; Matsumoto, T; Hidaka, E; Oguchi, H; Kanai, S; Oana, K; Yamauchi, K; Honda, T; Kawakami, Y
First case of bacteremia due to chromosome-encoded CfxA3-beta-lactamase-producing Capnocytophaga sputigena in a pediatric patient with acute erythroblastic leukemia. Eur J Med Res,31(13(3)):133-135 2008(Mar.) Author:Matsumoto, T; Matsubara M; Oana K; Kasuga E; Suzuki T; Hidaka E; Shigemura T; Yamauchi K; Honda T; Ota H; Kawakami Y
Sialic acid moiety of apolipoprotein E3 at Thr194 affects its interaction with β-amyloid1–42 peptides. Clin Chim Acta,388(1-2):123-129 2008(Feb.) Author:Sugano, M; Yamauchi K; Kawasaki K; Tozuka M; Fujita K; Okumura N; Ota H Abstract:Background: The interaction between apolipoprotein (apo) E and beta-amyloid (A beta) is associated with the development of Alzheimer's disease (AD); however, the details remain unknown. ApoE in cerebrospinal fluid is extensively sialylated, and sialylation of certain proteins are known to modulate biological function. We investigated the effects of a sialic acid moiety of apoE on the apoE-A interaction.
Methods: We prepared normal apoE3 and its mutant (Thr(194) -> Ala) and analyzed their interactions with A beta(1-42) by using the surface plasmon resonance (SPR) assay. In addition, we per-formed the SPR assay by using apoE-containing lipoproteins treated with neuraminidase. We also assessed the effect of the mutation on the interaction of apoE3 with liposomes.
Results: The binding avidity of the mutant apoE3(#) was approximately 50% that of normal apoE3 (p<0.0001). The binding avidity of the apoE-containing lipoproteins for A beta(1-42) reduced after neuraminidase treatment.
Conclusions: We suggest that AD development is controlled not only by the apoE isoforms but also by the posttranslational modifications in apoE, such as those in the sialic acid moieties, which are abundant in apoE derived from the brain. (c) 2007 Elsevier B.V All rights reserved.
Quantitative analysis of the effect of Helicobacter pylori on the expressions of SOX2, CDX2, MUC2, MUC5AC, MUC6, TFF1, TFF2, and TFF3 mRNAs in human gastric carcinoma cells. Scand J Gastroenterol,43:25-33 2008(Jan.) Author:Matsuda, K; Yamauchi, K; Matsumoto, T; Sano, K; Yamaoka, Y; Ota, H
Regulation of Interleukin-18 in Helicobacter Infection. J Immunol,180:1211-1217 2008(Jan.) Author:Yamauchi, K; Coi, Il-Ju; Lu, H; Ogiwara, H; Graham, DY; Yamaoka, Y
In vitro expression of β-thalassaemia gene (IVS I-1 G-->C) reveals complete inactivation of the normal 5’ splice site and alternative aberrant RNA splicing. Ann Clin Biochem,44:573-578 2007(Nov.) Author:Fujihara, N; Yamauchi, K; Hirota-Kawadobora, M; Ishikawa, S; Tozuka, M; Ishii, E; Katsuyama, T; Okumra, N; Taniguchi, S
Acquisition of loss of the wild-type NRAS locus with aggressive disease progression in a patient with juvenile myelomonocytic leukemia and a heterozygous NRAS mutation. Haematologica,92:1576-1578 2007(Nov.) Author:Matsuda, K; Nakazawa, Y; Sakashita, K; Shiohara, M; Yamauchi, K; Koike, K
Helicobacter pylori environmental interactions: effect of acidic conditions on H-pylori-induced gastric mucosal interleukin-8 production CELLULAR MICROBIOLOGY,9(10):2457-2469 2007(Oct.) Author:Il Ju Choi; Saori Fujimoto; Kazuyoshi Yamauchi; David Y. Graham; Yoshio Yamaoka Abstract:To explore the interactions between the host, environment and bacterium responsible for the different manifestations of Helicobacter pylori infection, we examined the effect of acidic conditions on H. pylori-induced interleukin (IL)-8 expression. AGS gastric epithelial cells were exposed to acidic pH and infected with H. pylori [wild-type strain, its isogenic cag pathogenicity island (PAI) mutant or its oipA mutant]. Exposure of AGS cells to acidic pH alone did not enhance IL-8 production. However, following exposure to acidic conditions, H. pylori infection resulted in marked enhancement of IL-8 production which was independent of the presence of the cag PAI and OipA, indicating that H. pylori and acidic conditions act synergistically to induce gastric mucosal IL-8 production. In neutral pH environments H. pylori-induced IL-8 induction involved the NF-kappa B pathways, the extracellular signal-regulated kinase (ERK)-> c-Fos/c-Jun -> activating protein (AP-1) pathways, JNK -> c-Jun -> AP-1 pathways and the p38 pathways. At acidic pH H. pylori-induced augmentation of IL-8 production involved markedly upregulated the NF-kappa B pathways and the ERK -> c-Fos -> AP-1 pathways. In contrast, activation of the JNK -> c-Jun -> AP-1 pathways and p38 pathways were pH independent. These results might explain the clinical studies in which patients with duodenal ulcers had higher levels of IL-8 in the antral gastric mucosa than patients with simple H. pylori gastritis.
Analysis of human serum lipoprotein lipid composition using maldi-tof mass spectrometry. Ann Clin Lab Sci,37:213-221 2007(Aug.) Author:Hidaka H; Hanyu N; Sugano M; Kawasaki K; Yamauchi K; Katsuyama T
A complex karyotype, including a three-way translocation generating a NUP98-HOXD13 transcript, in an infant with acute myeloid leukemia. Cancer Genet Cytogenet,15(176):137-143 2007(Jul.) Author:Hidaka, E; Tanaka, M; Matsuda, K; Ishikawa-Matsumura, M; Yamauchi, K; Sano, K; Honda; T. Wakui K; Yanagisawa, R; Nakazawa, Y; Sakashita, K; Shiohara, M; Ishii, E; Koike, K
Spontaneous improvement of hematologic abnormalities in patients having juvenile myelomonocytic leukemia with specific RAS mutations BLOOD,109(12):5477-5480 2007(Jun.) Author:Kazuyuki Matsuda; Akira Shimada; Nao Yoshida; Atsushi Ogawa; Akihiro Watanabe; Shuhei Yajima; Susumu Iizuka; Kazutoshi Koike; Fumio Yanai; Keiichiro Kawasaki; Masakatsu Yanagimachi; Akira Kikuchi; Yoshitoshi Ohtsuka; Eiko Hidaka; Kazuyoshi Yamauchi; Miyuki Tanaka; Ryu Yanagisawa; Yozo Nakazawa; Masaaki Shiohara; Atsushi Manabe; Seiji Kojima; Kenichi Koike Abstract:Of 11 children with juvenile myelomonocytic leukemia (JMML) carrying RAS mutations (8 with NRAS mutations, 3 with KRAS2 mutations), 5 had a profound elevation in either or both the white blood cells and spleen size at diagnosis. Three patients had no or modest hepatosplenomegaly and mild leukocytosis at presentation but subsequently showed a marked increase in spleen size with or without hematologic exacerbation, for which nonintensive chemotherapy was initiated. The other three patients with NRAS or KRAS2 glycine to serine substitution received no chemotherapy, but hematologic improvement has been observed during a 2- to 4-year follow up. In the third group, all hematopoletic cell lineages analyzed had the RAS mutations at the time of hematologic improvement, whereas DNA obtained from the nails had the wild type. Additionally, numbers of circulating granulocyte-macrophage progenitors were significantly reduced during the clinical course. Thus, some patients with JMMIL with specific RAS mutations may have spontaneously improving disease.
Transmission via the face is one route of methicillin-resistant Staphylococcus aureus cross-infection within a hospital AMERICAN JOURNAL OF INFECTION CONTROL,35(2):126-130 2007(Mar.) Author:Akane Kuramoto-Chikamatsu; Takayuki Honda; Takehisa Matsumoto; Mayumi Shiohara; Yoshiyuki Kawakami; Kazuyoshi Yamauchi; Yumiko Kato Abstract:Background: It is generally accepted that hospital personnel must not touch their faces, and indeed must not elevate their hands above the shoulder in the ward, because many bacteria including Staphylococcus aureus colonize the face. However, possible methicillin-resistant Staphylococcus aureus (MRSA) cross-infection by way of faces has not yet been properly examined.
Methods: One hundred and seventy-eight isolates from 159 inpatients from February 1999 to January 2000 were subjected to chromosomal DNA analysis by pulsed-held gel electrophoresis (PFGE).
Results: The 178 MRSA isolates were classified into 43 PEGE types. Cross-infection was more frequent in the Department of Neurology [average number of patients per PFGE pattern (n/PFGE) = 4.0] than in the other six nonsurgical departments (n/PFGE = 1.0-1.6), and more frequent in four surgical departments (n/PFGE = 2.0-4.5) than in the other eight (n/PFGE = 1.0-1.7). In neurology, patients' faces were more often touched for examination of the cranial nerves than in the other departments. In the above four surgical departments, organs in the face were chiefly operated upon, and the patients' faces were also touched for care before and after the operation.
Conclusions: Our study reveals the possibility of MRSA being transmitted by way of patients' faces in a hospital.
Quantitative Determination of Gland Mucous Cells-type Mucin Using a Monoclonal Antibody, HIK1083: Its Pathophysiological Changes in Human Gastric Juice. Clin Chim Acta,377:261-267 2007(Feb.) Author:Kubota, S; Yamauchi, K; Kumagai, T; Sugano, M; Kawasaki, K; Tozuka, M; Akamatsu, T; Matsuzawa, K; Sugiyama, A; Kurihara, M; Katsuyama, T; Ota, H
Evaluation of BacT/Alert 3D SA bottles for accurate detection of Mycobacteremia with special reference to Mycobacterium abscessus Eur J Med Res,12:43-46 2007(Jan.) Author:Kasuga, E; Matsumoto, T; Oana, K; Shiohara, M; Okabe, T; Yamauchi, K; Honda, T; Ota, H; Kawakami, Y
B:b interactions are essential for polymerization of variant fibrinogens with impaired holes ”a”. J Thromb Haemost,:2352-2359 2007(Jan.) Author:Okumura, N; Terasawa, F; Haneishi, A; Fujiwara, N; Hirota-kawadobora, M; Yamauchi, K; Ota, H; Lord, ST
TFF2 in gland mucous cell mucin in the mucous gel covering normal or damaged gastric mucosa using the Mongolian gerbil model. Scand J Gastroenterol,41:1390-1397 2006(Dec.) Author:Suzuki, K; Hayama, M; Nakamura, M; Yamauchi, K; Maruta, F; Miyagawa, S; Ota, H
A case of acute myelogenous leukemia with MLL-AF10 fusion caused by insertion of 5'MLL into 10p12, with concurrent 3'MLL deletion. Cancer Genet Cytogenet,17:124-130 2006(Nov.) Author:Matsuda, K; Hidaka, E; Ishida, F; Yamauchi, K; Makishima, H; Ito, T; Suzuki, T; Imagawa, E; Sano, K; Katsuyama, T; Ota, H
First isolation of Dysgonomonas mossii from intestinal juice of a patient with pancreatic cancer ARCHIVES OF MEDICAL RESEARCH,37(7):914-916 2006(Oct.) Author:Takehisa Matsumoto; Yoshiyuki Kawakami; Kozue Oana; Takayuki Honda; Kazuyoshi Yamauchi; Yukie Okimura; Mayumi Shiohara; Eriko Kasuga Abstract:Background. Dysgonomonas species were first designated in 2000. However, clinical infections due to this microorganism have rarely been described. Our aim was to present the first isolation of Dysgonomonas mossii from intestinal juice of a patient with pancreatic cancer.
Methods. Predominantly appearing grayish-white colonies grown on chocolate and sheep blood agar plates were characterized morphologically by: Gram stain, biochemically by automated instrument using Vitek II ID-GNB card together with commercially available kit systems, ID-Test HN-20 and API rapid ID 32A32A, and genetically by sequencing the 16S rRNA gene of the organism using a Taq DyeDeoxy Terminator Cycle Sequencing and a model 3 100 DNA sequencer instrument. The isolate was further characterized by antimicrobial susceptibility using MicroFast 4J Panels and additional biochemical and physiological properties.
Results. The isolate was finally identified as D. mossii from the findings of the morphological, cultural, and biochemical properties together with the comparative sequence of the 16S rRNA genes. The isolate was highly susceptible to many antibiotics but resistant to penicillins and cephems.
Conclusions. As D. mossii was rarely encountered in the clinical microbiology laboratory, it may be misidentified as an X-factor-dependent Haemophilus species due to its negative result for the porphyrin test. Accumulation of the case reports with the isolation of this species is expected to elucidate the infections due to D. mossii. The presence of D. mossii caused no significant clinical infection despite repeated isolations, as the patient had no conspicuous abdominal complaints. However, our report is a noteworthy and useful piece of information. (C) 2006 IMSS. Published by Elsevier Inc.
Analysis of fibrinogen variants at {gamma}387Ile shows that side-chain of {gamma}387 and the tertiary structure of {gamma}C terminal tail are important for not only assembly and secretion of fibrinogen but also lateral aggregation of protofibrils and XIII Blood,108:1887-1894 2006(Sep.) Author:Kani, S; Terasawa, F; Yamauchi, K; Tozuka, M; Okumura, N
Mechanism of IgA-albumin complex formation that affects the fructosamine assay. J Electrophoresis,50(2):19-23 2006(Jun.) Author:Fujita K; Kameko F; Kato Y; Fukushima M; Terasawa F; Sugano M; Yamauchi K; Sato H; Kameko M; Sakurabayashi I Abstract:We recently demonstrated glycation of monoclonal IgA and the presence of IgA-albumin complexes, but the mechanism of IgA-albumin complex formation was not clear. We isolated the IgA-albumin complexes from 5 IgA type M-proteinemia patients' sera. To elucidate the mechanism of IgA-albumin complex formation, we performed the dissociation assay of IgA-albumin complexes, the identification of albumin binding sites of monoclonal IgA using immunoelectrophoresis, western blotting and chromatography technologies. In all patients with IgA type M-proteinemia, the IgA-albumin complexes were dissociated by treated with 2-mercaptoethanol (2-ME), but not by treated with a strong acid as acetic acid or NaCl of high concentrations. Moreover, when the purified monoclonal IgA containing IgA-albumin complexes was digested with the IgA protease from Neisseria gonorrhoeae, no macro-albumin was demonstrated. It seems probable that albumin is bound to the monoclonal IgA molecule by covalent disulfide bonds, and that the binding site of albumin is located in near the hinge region of IgA molecule and involve the free SH group thought to be present in the α-chain.
A novel variant fibrinogen, deletion of B beta 111Ser in coiled-coil region, affecting fibrin lateral aggregation CLINICA CHIMICA ACTA,365(1-2):160-167 2006(Mar.) Author:N Okumura; F Terasawa; M Hirota-Kawadobora; K Yamauchi; K Nakanishi; S Shiga; S Ichiyama; M Saito; M Kawai; T Nakahata Abstract:Background: Functional fibrinogen concentration of a male infant showed < 0.50 g/l and we speculated this patient as a dysfibrinogenemia or hypofibrinogenemia.
Methods: We analyzed propositus and his parent by DNA sequencing and by thrombin-catalyzed fibrin polymerization for purified plasma fibrinogen.
Results: Although functional fibrinogen determinations based on Clauss method showed the marked discrepancy of values among 3 sets of reagent and analyzer, we found a novel heterozygous variant fibrinogen, Kyoto IV, caused by 3-bp deletion in B beta-chain gene corresponding to the deletion of 111Ser located in coiled-coil region. We suggested that the discrepancy of fibrinogen values among 3 assays was caused by the difference in NaCl concentration in reagents for determination and analyzed the polymerization under the conditions of various NaCl concentrations. Although under normal physiological conditions Kyoto IV fibrinogen augmented the polymerization as compared with normal control, in 0.21 mol/l NaCl Kyoto IV fibrinogen showed abruptly impaired polymerization curve compared with normal control.
Conclusion: Variant fibrinogen, B beta Delta 111Ser, showed augmented lateral aggregation under normal physiological conditions and the residue located in coiled-coil region, B beta 111Ser, plays an important role in the lateral aggregation. (c) 2005 Elsevier B.V. All rights reserved.
α1,4-N-acetylglucosaminyltransferase mRNA for detection of pancreatic cancer. Cancer Sci,97:119-126 2006(Feb.) Author:Ishizone, S; Yamauchi, K; Kawa, S; Suzuki, T; Shimizu, F; Harada, O; Sugiyama, A; Miyagawa, S; Fukuda, M; Nakayama, J
New enzymatic assay using phospholipase D to measure total calcium in serum. Clin Chem 2005(Jun.) Author:Sugano M; Yamauchi K; Sugano K; Kawasaki K; Tozuka M; Katsuyama T; Soya H; Tanaka T; Imamura S; Nomoto S
Functional analysis of recombinant B beta 15C and B beta 15A fibrinogens demonstrates that B beta 15G residue plays important roles in FPB release and in lateral aggregation of protofibrils JOURNAL OF THROMBOSIS AND HAEMOSTASIS,3(5):983-990 2005(May) Author:M Hirota-Kawadobora; S Kani; F Terasawa; N Fujihara; K Yamauchi; M Tozuka; N Okumura Abstract:Background and objectives: Analysis of dysfibrinogens has improved our understanding of molecular defects and their effects on the function of intact fibrinogen. To eliminate the influence of plasma heterozygous molecules, we synthesized and analyzed recombinant-variant fibrinogens. Methods: We synthesized two recombinant-variant fibrinogens with a single amino acid substitution at the 15Gly residue in the Bβ-chain: namely, Bβ 15Cys and Bβ 15Ala. Results: Western blotting analysis of purified fibrinogen revealed the existence of a small amount of a dimeric form only for Bβ 15Cys fibrinogen. For Bβ 15Cys fibrinogen, functional analysis indicated (a) no thrombi n-catalyzed fibrinopeptide B (FPB) release and (b) markedly impaired lateral aggregation in thrombin- and reptilase-catalyzed fibrin polymerizations. For Bβ 15Ala fibrinogen, such analysis indicated slight impairments of both thrombin-catalyzed FPB release and lateral aggregation in thrombin-catalyzed fibrin polymerization, but nearly normal lateral aggregation in reptilase-catalyzed fibrin polymerization. These impaired lateral aggregations were accompanied by thinner fibrin fiber diameters (determined by scanning electron microscopy of the corresponding fibrin clots). Conclusion: We conclude that a region adjacent to Bβ 15Gly plays important roles in lateral aggregation not only in desA fibrin polymerization, but also in desAB fibrin polymerization, and we speculate that the marked functional differences between Bβ 15A and Bβ 15C fibrinogens in FPB release and fibrin polymerization might not only be due to the presence of a substituted cysteine residue in BP I 5C fibrinogen, but also to the existence of disulfide-bonded forms. Finally, our data indicate that the Bβ 15Gly residue plays important roles in FPB release and lateral aggregation of protofibrils.
Roles of virD4 and cagG genes in the cag pathogenicity island of Helicobacter pylori using a Mongolian gerbil model. Gut 2005(May) Author:Saito H; Yamaoka Y; Ishizone S; Maruta F; Sugiyama A; Graham DY; Yamauchi K; Ota H; Miyagawa S
Natural history of gastric mucosal cytokine expression in Helicobacter pylori gastritis in Mongolian gerbils. Infect Immun 2005(Apr.) Author:Yamaoka Y; Yamauchi K; Sugiyama A; Ishizone S; Grahama DY; Maruta F; Murakami M; Katsuyama T
Monitoring of hematopoietic chimerism by short tandem repeats, and the effect of CD selection on its sensitivity. Clin Chem 2004(Dec.) Author:Matsuda K; Yamauchi K; Tozuka M; Suzuki T; Sugano M; Hidaka E; Sano K; Katsuyama T
Degradation of pre-beta-high density lipoproteins and their binding activity to human blood monocytes. Ann Clin Lab Sci 2004(Jul.) Author:Nakabayashi T; Yamauchi K; Sugano M; Sano K; Tozuka M; Hidaka H
Characterization of factor XII Tenri, a rare CRM-negative factor XII deficiency. Ann Clin Lab Sci 2004(Mar.) Author:Fujihara N; Tozuka M; Yamauchi K; Ueno I; Urasawa N; Ishikawa S; Hirota-Kawadobora M; Okumura N; Hidaka H; Katsuyama T
Purification and measurement of HDL3-binding proteins in human peripheral blood mononuclear cells. Ann Clin Lab Sci 2003(Jul.) Author:Hidaka H; Tozuka M; Yamauchi K; Ohta H; Nakayama J; Katsuyama T
Characterization of triglyceride rich lipoproteins with very light density by ultracentrifugation and agarose gel electrophoresis using triglyceride- and cholesterol-staining. Ann Clin Lab Sci 2003(Mar.) Author:Hidaka H; Tozuka M; Meyer B; Yamauchi K; Sugano M; Nakabayashi T; Katsuyama T
‘Severe’ Prader-Willi syndrome with a large deletion of chromosome 15 due to an unbalanced t(15,22)(q14;q11.2) translocation. Clin Genet 2003(Jan.) Author:Matsumura M; Kubota T; Hidaka E; Wakui K; Kadowaki S; Yamauchi K; Herzing LB; Nurmi EL; Sutcliffe JS; Fukushima Y; Katsuyama T
Novel beta-thalassemia trait (IVS I-1 G-->C) in a Japanese family. Am J Hematol 2003(Jan.) Author:Fujihara N; Tozuka M; Ueno I; Yamauchi K; Nakagoshi R; Ishikawa S; Hirota M; Ishii E; Katsuyama T
Internalization of beta-amyloid causes downregulation of apolipoprotein E mRNA expression in neuroblastoma cells. Ann Clin Lab Sci 2003(Jan.) Author:Yamauchi K; Tozuka M; Hidaka E; Ueno I; Matsuda K; Katsuyama T
血清亜鉛値による亜鉛欠乏症システムの現況：その欠落部への提言 Biomedical Research on Trace Elements 2003 Author:野本昭三; 山内一由; 中林徹雄
Predominant apolipoprotein J exists as lipid-poor mixtures in cerebrospinal fluid. Ann Clin Lab Sci 2002(Sep.) Author:Suzuki T; Tozuka M; Yamauchi K; Sugano M; Nakabayashi T; Okumura N; Hidaka H; Katsuyama T; Higuchi K
DNA microarray analysis of T-cell type lymphoproliferative disease of granular lymphocytes. Br J Hematol 2002(Aug.) Author:Makishima H; Ishida F; Ito T; Kitano K; Ueno S; Ohmine K; Yamashita Y; Ota J; Ota M; Yamauchi K; Mano H
Quantitative RT-PCR analysis demonstrates that synthesis of the recombinant fibrinogen is dependent on the transcription and synthesis of gamma-chain. Clin Chim Acta 2002(May) Author:Hirota M; Tozuka M; Yamauchi K; Hidaka E; Ueno I; Sugano M; Terasawa F; Okumura N; Katsuyama T; Sigematsu H
t(8;21;14)(q22;q22;q24) is anovel variant of t(8;21) with chimeric transcripts of AML1- ETO in acute myelogenous leukemia. Cancer Genet Cytogenet 2002(Jan.) Author:Ishida F; Ueno M; Tanaka H; Makishima H; Suzawa K; Hosaka S; Hidaka E; Ishikawa M; Yamauchi K; Kitano K; Kiyosawa K
Isoform-specific effect of apolipoprotein E on endocytosis of beta-amyloid in cultures of neuroblastoma cells. Ann Clin Lab Sci. 2002(Jan.) Author:Yamauchi K; Tozuka M; Hidaka H; Nakabayashi T; Sugano M; Katsuyama T
Characterization of an apolipoprotein E3 variant (Arg 145-->His) associated with mild hypertriglyceridemia. Ann Clin Lab Sci 2001(Apr.) Author:Hidaka H; Tozuka M; Hidaka E; Yamauchi K; Ota H; Katsuyama T
Evaluation of the fluorescent image analyzer “Image Titer” for quantitative analysis of antinuclear antibodies. Am J Clin Pathol 2001(Mar.) Author:Nakabayashi T; Kumagai T; Yamauchi K; Kuramoto A; Fujita K; Hidaka H; Tozuka M
Effect of apolipoprotein AII on the interaction of apolipoprotein E with beta-amyloid: some apo(E-AII) complexes inhibit the internalization of beta-amyloid in cultures of neuroblastoma cells. J Neurosci Res. 2000(Nov.) Author:Yamauchi K; Tozuka M; Hidaka H; Nakabayashi T; Sugano M; Kondo Y; Nakagawara A; Katsuyama T
Novel BCR-ABL transcript containing an intronic sequence insert in a patient with Philadelphia-positive acute lymphoblastic leukaemia. Br J Haematol 2000(Sep.) Author:Hirota M; Hidaka E; Ueno I; Ishikawa M; Asano N; Yamauchi K; Ishida F; Tozuka M; Katsuyama T
Analysis of hemoglobin and globin chain variants by a commonly used capillary isoelectric focusing method. Electrophoresis 2000(Aug.) Author:Sugano M; Hidaka H; Yamauchi K; Nakabayashi T; Higuchi Y; Fujita K; Okumura N; Ushiyama Y; Tozuka M; Katsuyama T
Higher avidity binding of apolipoprotein (E-AII) complex than of apolipoprotein E monomer to beta-amyloid. J Neurosci Res 1999(Oct.) Author:Yamauchi K; Tozuka M; Nakabayashi T; Sugano M; Hidaka H; Kondo Y; Katsuyama T
Characterization of apolipoprotein E-containing lipoproteins in cerebrospinal fluid: effect of phenotype on the distribution of apolipoprotein E. Clin Chem 1999(Sep.) Author:Yamauchi K; Tozuka M; Hidaka H; Hidaka E; Kondo Y; Katsuyama T
Genetic analyses in homozygous and heterozygous variants of lactate dehydrogenase-B (H) subunit - LD-B Matsumoto I and II (LD-B W323R) CLINICA CHIMICA ACTA,287(1-2):163-171 1999(Sep.) Author:N Okumura; F Terasawa; Ueno, I; K Oki; K Yamauchi; H Hidaka; M Tozuka; M Okura; T Katsuyama
Apolipoprotein E in cerebrospinal fluid: relation to phenotype and plasma apolipoprotein E concentrations. Clin Chem 1999(Apr.) Author:Yamauchi K; Tozuka M; Nakabayashi T; Sugano M; Hidaka H; Kondo Y; Katsuyama T
Characterization of hypertriglyceridemia induced by L-asparaginase therapy for acute lymphoblastic leukemia and malignant lymphoma. Ann Clin Lab Sci 1997 Author:Tozuka M; Yamauchi K; Hidaka H; Nakabayashi T; Okumura N; Katsuyama T
講演・口頭発表等 Balance capability and smell identification are associated with cognitive function in middle-aged persons with type 2 diabetes 56th Annual Meeting of the European-Association-for-the-Study-of-Diabetes (EASD) 2020(Sep. 21) Presenter:Suzuki, H; Midorikawa, M; Suzuki, Y; Sato, H; Nemoto, K; Yamauchi, K; Sugano, Y; Osaki, Y; Iwasaki, H; Sekiya, M; Yahagi, N; Hada, Y; Arai, T; Shimano, H
研究者総覧
