Biomedical subjects

I Yamaguchi

Publications and source records attributed to I Yamaguchi.

At least 19 recordsLinked to original sources

PubMed2007

Tissue factor expression as a possible determinant of thromboembolism in ovarian cancer.

Ovarian cancer, and clear cell carcinoma in particular, reportedly increases the risk of venous thromboembolism (VTE). However, the mechanisms remain unclear. Tissue factor (TF) supposedly represents a major factor in the procoagulant activities of cancer cells. The present study examined the involvement of TF expression in VTE for patients with ovarian cancer. Subjects comprised 32 consecutive patients (mean age 49.8 years) with histologically confirmed ovarian cancer. Presence of VTE was examined using a combination of clinical features, D-dimer levels and venous ultrasonography. Immunohistochemical analysis was used to evaluate TF expression into 4 degrees. Venous thromboembolism was identified in 10 of the 32 patients (31%), including five of the 11 patients with clear cell carcinoma. Tissue factor expression was detected in cancer tissues from 24 patients and displayed significant correlations with VTE development (P=0.0003), D-dimer concentration (P=0.003) and clear cell carcinoma (P<0.05). Multivariate analysis identified TF expression as an independent predictive factor of VTE development (P<0.05). Tissue factor (TF) expression is a possible determinant of VTE development in ovarian cancer. In particular, clear cell carcinoma may produce excessive levels of TF and is more likely to develop VTE.

Adolescent↗

PubMed2006

Cloning and characterization of six highly similar endo-1,3-beta-glucanase genes in hexaploid wheat.

Pathogenesis-related (PR) proteins are often used as a marker of plant defense reactions. Some endo-1,3-beta-glucanase (Gns) genes encode proteins that belong to the PR protein family 2 (PR-2). Although the number of homologous family member genes is significantly greater in hexaploid wheat (Triticum aestivum L.) compared to other model plants, earlier studies did not evaluate the possible contribution of their homologs to hybridization signals in Northern blot analysis. In this study, we have examined whether routine transcriptional analyses of a PR gene is of high reliability or not by isolating six highly similar Gns genes (TaGlb2a, TaGlb2b, TaGlb2c, TaGlb2d, TaGlb2e, and TaGlb2f) and characterizing their expression patterns in detail. While TaGlb2b was shown to be a PR-2 gene, transcription of TaGlb2c and TaGlb2d was not induced upon infection with either powdery mildew (Erysiphe graminis) or head blight (Fusarium graminearum) pathogens; their transcripts were most abundant in healthy spikes (lemmas and in particular paleae). Therefore, in some cases, the conventional analyses do not necessarily provide accurate information on expression pattern of a PR gene in hexaploid wheat. This is also the first report of wheat genes that are specifically expressed in lemma/palea tissues of flowering spikelets.

Base Sequence↗

PubMed2005

Secretion of bacterial xenobiotic-degrading enzymes from transgenic plants by an apoplastic expressional system: an applicability for phytoremediation.

In search of an effective method for phytoremediation of wastewater contaminated with organic compounds, we investigated the application of an apoplastic expressional system that secretes useful bacterial enzymes from transgenic plants into hydroponic media through the addition of a targeting signal. We constructed transgenic Arabidopsis expressing the aromatic-cleaving extradiol dioxygenase (DbfB), which degrades 2,3-dihydroxybiphenyl (2,3-DHB), and transgenic tobacco expressing haloalkane dehalogenase (DhaA), which catalyzes hydrolytic dechlorination of 1-chlorobutane (1-CB). Although crude leaf extracts of transgenic plants expressing cytoplasm-targeted degradative enzymes showed higher activity than did those from transgenic plants expressing apoplast-targeted enzymes, the hydroponic media of the latter showed 23.2 times (DbfB) and 76.4 times (DhaA) higher activity than plants containing the cytoplasm-targeted enzymes. Addition of crystalline 2,3-DHB to 100 mL of the hydroponic medium of transgenic or wild-type seedlings revealed that only medium from the transgenic Arabidopsis expressing apoplast-targeted DbfB showed rapid ring cleavage of 2,3-DHB. Transgenic tobacco expressing apoplast-targeted DhaA also resulted in the accumulation of the dehalogenation product 1-butanol in the hydroponic medium and showed a higher tolerance to 1-CB than wild-type or transgenic plants expressing cytoplasm-targeted DhaA. These results demonstrate the usefulness of the apoplastic expression of bacterial recombinant proteins in phytoremediation.

Actinomycetales↗

PubMed2004

Bayesian classification of myocardial excitation abnormality using magnetocardiogram maps for mass screening.

We propose a novel classification method based on the Bayes rule to utilize the magnetocardiogram (MCG) in noninvasive mass screening. The cardiac excitation is directly tracked by maps of the MCG field generated by myocardial excitation current through the excited wave front. To adopt the characteristics of the excited wave fronts as a parameter for the Bayes theorem, we developed a parameterization procedure that consists of a two-dimensional wavelet approximation and a cluster analysis of magnetic field maps. With the parameter determined by this procedure, the probability of a subject to belong to a disease group or to the normal group is estimated by the Bayes theorem. The subject is classified into the group of the highest probability. We applied the proposed method to ST-T period of MCG data of 6 old myocardial infarction (OMI) patients and 15 normal controls. The method showed sensitivity of 83%; specificity, 100%; positive predictive value, 100%; and negative predictive value, 94% in the classification of OMI patients and normal controls. The processing time is less than 5 seconds per one subject. It suggests a possible application of the proposed method in mass screening of abnormal MCG patterns.

Bayes Theorem↗

PubMed2004

Pattern of N gene-mediated systemic hypersensitive response and turnover of viral replicase protein in tobacco.

We examined the pattern of the N gene-mediated systemic hypersensitive response (HR), which was induced by tobacco mosaic virus upon temperature shift, and analyzed the distribution of the coat protein and the virus-encoded 126 kDa replicase protein (126 K protein) by immunoblot analysis. In the middle- and lower-positioned leaves, HR occurred in the advancing edge of the infected area, where we detected both the coat protein and the 126 K protein. In the areas between the main vein and the advancing edge of these leaves, we observed no HR and did not detect 126 K protein, though virus was present in these areas. In the upper-positioned mosaic leaves, patterns of the HR were different depending on the leaf age. In these mosaic leaves, the mechanism preventing the virus from invading dark green tissue seemed to be broken down in 8-14 days old leaves, and HR was observed only in the tissue just invaded by the virus, where we detected the 126 K protein. Those results suggested that the viral 126 K protein was present when the viral replication was taking place, and easily degraded when the amount of the virus was saturated in the cells.

Capsid Proteins↗

PubMed2003

Noninvasive, direct visualization of macro-reentrant circuits by using magnetocardiograms: initiation and persistence of atrial flutter.

AIMS: We analysed the cardiac magnetic fields on the body surface to visualize electrical currents noninvasively during reentrant arrhythmias. METHODS AND RESULTS: Seven patients with counterclockwise atrial flutter (AFL) were studied during 17 episodes of AFL using 64-channel magnetocardiograms (MCGs) and electrophysiological study. Eight of the episodes were paroxysmal AFL, in which MCGs were recorded from the time of spontaneous onset to the time of termination. We constructed iso-magnetic field maps of the tangential components and produced MCG animations. With respect to AFL initiation, an atrial premature complex induced AFL. Prior to the initiation of AFL, atrial fibrillation (AF) transiently occurred. The cardiac magnetic fields revealed a single peak during sinus rhythm or with premature complexes but a disorganized pattern during AF. When AF transformed to AFL, the magnetic fields changed from a disorganized pattern to a single peak at first and then evolved to a circular pattern. During persistent AFL, the magnetic source moved in a counterclockwise circuit. CONCLUSION: MCG animation can be used to visualize the sequence in which a premature complex transforms sinus rhythm to AFL via AF. Our findings indicate that MCGs can be used to identify noninvasively the mechanisms responsible for atrial tachyarrhythmias.

Atrial Fibrillation↗

PubMed2003

Microstructure analysis of calcium phosphate formed in tendon.

The surface of soft tendon tissue has been modified using calcium phosphate in order for the tendon to directly connect with hard bone and reconstruct an injured ligament. Calcium phosphate was coated onto the tendon in a soaking process using alternating a CaCl(2) (200 mM) and a Na(2)HPO(4) (120 mM) solution. According to SEM/EDX observations, calcium phosphate was formed, not only on the tendon surface, but also inside the tendon tissue. When the tendon was treated with seven soaking cycles, calcium phosphate was detected between 0-500 microm from the tendon surface. According to TEM observations, the crystal morphology of calcium phosphate depends on the distance from the surface. Hydroxyapatite crystals were observed near the surface, while octa-calcium phosphate crystals could be observed further from the surface, thus at initial soaking. The crystals were formed on collagen fibrils in spaces between the collagen fibrils with the c-axes of the crystals aligned parallel with the collagen fibrils. This finding suggests Ca(2+) ions to interact with the tendon surface, most probably with the carboxyl functional groups of collagen, and subsequently forming nucleation centers for the crystals.

Journal Article↗

PubMed2001

Apolipoprotein E inhibits the depolymerization of beta 2-microglobulin-related amyloid fibrils at a neutral pH.

beta 2-Microglobulin-related (A beta 2M) amyloidosis is a common and serious complication in patients on long-term hemodialysis, and beta 2-microglobulin (beta 2-m) is a major structural component of A beta 2M amyloid fibrils. Fluorescence spectroscopic analysis with thioflavin T and electron microscopic study revealed that A beta 2M amyloid fibrils readily depolymerize into monomeric beta 2-m at a neutral to basic pH. Circular dichroism analysis revealed that soon after the initiation of the depolymerization reaction at pH 7.5, the characteristic spectrum of beta 2-m in A beta 2M amyloid fibrils changes to resemble that of monomeric beta 2-m at pH 7.5. Apolipoprotein E (apoE), a representative amyloid-associated protein, formed a stable complex with A beta 2M amyloid fibrils and inhibited the depolymerization of A beta 2M amyloid fibrils dose-dependently in a range of 0--10 microM. These results showed that apoE could enhance the deposition of amyloid fibrils in vivo, possibly by binding directly to the surface of the fibrils and stabilizing the conformation of beta 2-m in the fibrils.

Amyloid beta-Peptides↗

PubMed2001

Novel molecular mechanism of increased myocardial endothelin-1 expression in the failing heart involving the transcriptional factor hypoxia-inducible factor-1alpha induced for impaired myocardial energy metabolism.

BACKGROUND: Hypoxia-inducible factor (HIF)-1alpha is an important transcriptional factor that activates the gene expression of glycolytic enzymes, which are activated as compensation for impaired beta-oxidation of fatty acid in the failing heart. We reported that cardiac endothelin (ET)-1 expression is markedly increased in heart failure. The mechanism, however, is unknown. Because we found an HIF-1alpha binding site in the 5'-promoter region of the ET-1 gene, we hypothesized that HIF-1alpha is involved in this mechanism. METHODS AND RESULTS: In rat cardiomyocytes, luciferase assay and electrophoretic mobility shift assay showed that HIF-1alpha transcriptionally activates ET-1 gene expression by direct interaction with the predicted DNA binding site in the 5'-promoter region. HIF-1alpha mRNA and ET-1 mRNA in the failing heart increased during the aggravation of heart failure in vivo in animal models, ie, rats with myocardial infarction and hamsters with cardiomyopathy. In cultured cardiomyocytes treated with a mitochondrial inhibitor, HIF-1alpha mRNA and ET-1 mRNA were markedly increased with activated glycolysis, and antisense oligonucleotide for HIF-1alpha largely inhibited the increased gene expression of ET-1. CONCLUSIONS: The present study revealed a novel molecular mechanism of upregulation of myocardial ET-1 in heart failure, indicating that induction of HIF-1alpha to stimulate glycolysis as an adaptation in heart failure against impaired energy metabolism alternatively causes an elevation of cardiac ET-1 gene expression as a maladaptation.

Adenosine Triphosphate↗

PubMed2001

Preparation and microstructure analysis of chitosan/hydroxyapatite nanocomposites.

Chitosan/hydroxyapatite (HAp) composites with a homogeneous nanostructure have been prepared by a co-precipitation method. According to TEM observations, HAp crystallites in the composites formed elliptic aggregations 230 nm in length and 50 nm in width. The typical length of the aggregations corresponded approximately to that of a chitosan molecule. The size of the constituent HAp crystallites was found to be predominantly 30 nm in length and 10 nm in width, and the c-axes were well aligned in parallel with the chitosan molecules in the respective aggregations. The growth of the HAp crystallites is considered to occur at nucleation sites, most probably forming the complexes with amino groups on chitosan with calcium ions. The compact composites obtained have been found to be mechanically flexible, and this flexibility has been improved further by heating at 120 degrees C for 20 min in an autoclave with saturated steam pressure.

Biocompatible Materials↗

PubMed2001

Characterization of mutations in the two-component histidine kinase gene that confer fludioxonil resistance and osmotic sensitivity in the os-1 mutants of Neurospora crassa.

Osmotic-sensitive (os-1) mutant alleles in Neurospora crassa exhibit resistance to dicarboximides, aromatic hydrocarbons and phenylpyrroles. We have previously reported that the os-1 mutants can be classified into two groups based on their resistance to fungicides and osmotic stress: type I, which are highly resistant to iprodione and fludioxonil but moderately sensitive to osmotic stress, and type II, which are highly sensitive to osmotic stress but moderately resistant to fungicides. To explain the mechanism of resistance to these fungicides, we cloned and sequenced the mutant os-1 genes that encode putative osmo-sensing histidine kinase. Within the os-1 gene product (Os1p), the type I strains, NM233t and Y256M209, carried a stop codon at amino acid position 308 and a frameshift at amino acid position 294, respectively. These mutation sites were located on the upstream of histidine kinase and the response regulator domains of Os1p, strongly suggesting that type I strains are null mutants. The null mutants, NM233t and Y256M209, were highly resistant to iprodione and fludioxonil; thus Os1p is essential for these fungicides to express their antifungal activity. The amino acid changes in Os1p, 625Pro from Leu, 578Val from Ala, and 580Arg from Gly were found in the type II strains, M16, M155-1 and P5990, respectively. Os1p is novel in having six tandem repeats of 90 amino acids in the N terminal. Each amino acid change of the type II strains was located on the fifth unit of six tandem repeats. Type II strains with single amino acid changes were more sensitive to osmotic stress than the null mutants (type I), indicating that the amino acid repeats of Os1p were responsible for an important function in osmo-regulation.

Amino Acid Sequence↗

PubMed2001

A novel transposon tagging element for obtaining gain-of-function mutants based on a self-stabilizing Ac derivative.

A novel tagging system AcREH, designed for obtaining gain-of-function mutations, was prepared on the basis of a self-stabilizing Ac transposon derivative. The transposable element, DsAT, was constructed in a way that it can activate transcription of neighboring genes by two 35S promoters and/or by four tandem repeats of the enhancer fragment of this promoter. DsAT revealed somatic excision in the first generation of the tobacco transformants. The element exhibited germinal excision to the next generation, as demonstrated by PCR and Southern hybridization analysis. In spite of the structure of the element, which may inhibit the expression of the transposase gene, the frequency of germinal excision was comparable to or higher than those so far reported, suggesting the applicability of the element for gene tagging.

Base Sequence↗

PubMed2001

Photocatalytic deposition of a gold nanoparticle onto the top of a SiN cantilever tip.

We propose a new technique for depositing a gold nanoparticle onto the tip of a dielectric support. We employed the photocatalytic effect of titanium dioxide for the deposition. When the titanium dioxide immersed in a solution including gold ions is subject to optical exposure, the excited electrons in the conduction band reduce gold ions into gold metal. Illumination by an evanescent wave generated with a total reflection configuration limits the deposition region to the very tip. In experiments we obtained 100-300 nm gold particles on SiN cantilever tips for atomic force microscopes. The contrast of evanescent interference fringes measured by a near-field scanning optical microscope with this gold nanoparticle probe has proved to be higher than that with a non-deposited SiN probe by a factor of 1.5.

Journal Article↗

PubMed2001

Probenazole induces systemic acquired resistance in Arabidopsis with a novel type of action.

Probenazole (PBZ; 3-allyloxy-1,2-benzisothiazole-1,1-dioxide), which is the active ingredient in Oryzemate, has been used widely in Asia to protect rice plants against the rice blast fungus Magnaporthe grisea. To study PBZ's mode of action, we analyzed its ability, as well as that of its active metabolite 1, 2-benzisothiazol-3 (2H)-one 1,1-dioxide (BIT) to induce defense gene expression and resistance in Arabidopsis mutants that are defective in various defense signaling pathways. Wild-type Arabidopsis treated with PBZ or BIT exhibited increased expression of several pathogenesis-related genes, increased levels of total salicylic acid (SA), and enhanced resistance to the bacterial pathogen Pseudomonas syringae pv. tomato DC 3000 and the oomycete pathogen Peronospora parasitica Emco5. The role of several defense signaling hormones, such as SA, ethylene and jasmonic acid (JA), in activating resistance following PBZ or BIT treatment was analyzed using NahG transgenic plants and etr1-1 and coi1-1 mutant plants, respectively. In addition, the involvement of NPR1, a key component in the SA signaling pathway leading to defense responses, was assessed. PBZ or BIT treatment did not induce disease resistance or PR-1 expression in NahG transgenic or npr1 mutant plants, but it did activate these phenomena in etr1-1 and coi 1-1 mutant plants. Thus SA and NPR1 appear to be required for PBZ- and BIT-mediated activation of defense responses, while ethylene and JA are not. Furthermore, our data suggest that PBZ and BIT comprise a novel class of defense activators that stimulate the SA/NPR1-mediated defense signaling pathway upstream of SA.

Arabidopsis↗

PubMed2001

Differential expression of adenylyl cyclases in the rat nephron.

BACKGROUND: Adenylyl cyclases (ACs) are a family of enzymes that catalyze the formation of the second-messenger cyclic adenosine 3',5'-monophosphate (cAMP). At least nine isoforms of AC have been cloned. These isoforms differ in their tissue distribution and basal activity. AC isoforms also differ in their capacity to be stimulated or inhibited by G protein alpha(i), alpha(s) and beta/gamma subunits, protein kinase C, and intracellular calcium. The distribution of ACs in the kidney is only partially known, although it is known that ACs play important roles in kidney signal transduction. Several receptors are known to couple to AC, but their linkage to individual AC isoforms in the kidney is not known. METHODS: This study investigated the tissue distribution of AC isoforms along the nephron of Wistar-Kyoto rats using reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and immunoblotting. RESULTS: While AC VI and IX mRNA were found in all nephron segments, there was no expression of AC VIII mRNA. ACs II through V and VII mRNA were variably found in specific nephron segments. mRNA for AC isoforms II, III, VI, VII, and IX were expressed in renal proximal tubules. All of the AC isoforms studied, except VIII, were found in glomeruli. Immunoblotting and immunohistochemistry confirmed the mRNA results. AC isoforms II, III, IV, and IX were expressed in luminal rather than in basolateral membranes. However, immunohistochemical studies were not feasible for the other isoforms that could be expressed in basolateral membranes. CONCLUSION: Knowledge of the distribution of ACs may help establish the linkage between receptors and specific AC isoforms and define their functions.

Adenylyl Cyclases↗

PubMed2001

Endothelin-1 production is enhanced by rotenone, a mitochondrial complex I inhibitor, in cultured rat cardiomyocytes.

In chronic heart failure and acute myocardial infarction, the tissue level of endothelin (ET)-1 in the heart, as well as its plasma level, has been reported to increase markedly. There is, however, little information about what in these pathologic conditions leads to increased production of ET-1, and which type of cell in the heart produces ET-1. We examined the mRNA and peptide expression of ET-1 using cultured rat neonatal cardiomyocytes, in which mitochondrial dysfunction was induced by rotenone, a mitochondrial respiratory chain complex I inhibitor, because one of the common features in failing or ischemic hearts is an alteration in energy metabolism due to mitochondrial dysfunction. Rotenone increased glucose use by the culture cells within 12 h of addition without affecting cell viability, and depressed the mitochondrial membrane potential after 72 h, indicating the induction of mitochondrial dysfunction in cardiomyocytes. Rotenone induced significant increase in the expression level of mRNA for ET-1 within 1 h of addition. In accordance with this finding, immunoreactive ET-1 in culture medium increased 3 times after 24 h of incubation, suggesting active secretion of ET-1 from cultured cells treated with rotenone. Immunocytochemical analysis verified significant increase of ET-1 peptide in cardiomyocytes, confirming the production of ET-1 by cardiomyocytes. These results suggest that derangement of mitochondrial function in cardiomyocytes itself could lead to the increased production of ET-1 in cardiomyocytes, and that this mechanism may contribute to the increased production of ET-1 in failing and ischemic hearts.

Animals↗

PubMed2001

[A clinical study on patients detected Pasteurella multocida from sputum].

We reported ten cases, (four female and six male), whose sputum cultures positive for Pasteurella multocida from 1990 to 2000. In the past eleven years increasing numbers of cases have appeared in our hospital. The majority of the cases with P. multocida possessed some underlying pulmonary diseases (seven cases, 70%), inactive lung tuberculosis or bronchiectasis. There were compromised hosts such as high ages person, steroids dependent person and diabetes mellitus patients. P. multocida was almost susceptible to antibioticus (penicillin and cephalosporins), although some erythromycin resistant strains were identified. The cats' oral cavities in our two cases were cultured and P. multocida were isolated. In our survey the prevalence of this organism is as high as 85% in cats. Our data suggests that patients who are in the high infection risk category are easily infected to P. multocida.

Adolescent↗

PubMed2001

Impaired renal D(1)-like and D(2)-like dopamine receptor interaction in the spontaneously hypertensive rat.

D(1)-like (D(1), D(5)) and D(2)-like (D(2), D(3), D(4)) dopamine receptors interact in the kidney to produce a natriuresis and a diuresis. Disruption of D(1) or D(3) receptors in mice results in hypertension that is caused, in part, by a decreased ability to excrete an acute saline load. We studied D(1)-like and D(2)-like receptor interaction in anesthetized spontaneously hypertensive rats (SHR) by the intrarenal infusion of Z-1046 (a novel dopamine receptor agonist with rank order potency of D(3)> or =D(4)>D(2)>D(5)>D(1)). Z-1046 increased glomerular filtration rate (GFR), urine flow, and sodium excretion in normotensive Wistar-Kyoto rats but not in SHRs. The lack of responsiveness to Z-1046 in SHRs was not an epiphenomenon, because intrarenal cholecystokinin infusion increased GFR, urine flow, and sodium excretion to a similar extent in the two rat strains. We conclude that renal D(1)-like and D(2)-like receptor interaction is impaired in SHRs. The impaired D(1)-like and D(2)-like receptor interaction in SHRs is not caused by alterations in the coding sequence of the D(3) receptor, the D(2)-like receptor expressed in rat renal tubules that has been shown to be involved in sodium transport. Because the diuretic and natriuretic effects of D(1)-like receptors are, in part, caused by an interaction with D(2)-like receptors, it is possible that the decreased Z-1046 action in SHRs is secondary to the renal D(1)-like receptor dysfunction in this rat strain.

Animals↗