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Biomedical subjects

M Yamaguchi

Publications and source records attributed to M Yamaguchi.

At least 505 records · Page 28Linked to original sources

High-Temperature In Situ Straining Experiments in the High-Voltage Electron Microscope.

: Design rules are described here for high-temperature straining stages for transmission electron microscopy. Temperatures above 1000 degreesC can be attained by electron bombardment of the specimen grips. Thermal equilibrium can be reached in a short time by carrying off the heat by water cooling. Some applications of this stage are described. Ferroelastic deformation was observed at 1150 degreesC in t' and partially stabilized zirconia, which changes the microstructure for successive dislocation plasticity. In the oxide-dispersion-strengthened alloy INCOLOY MA 956, dislocations are impeded by oxide particles and move smoothly between the particles. At high temperatures, both the resting and traveling times control the average dislocation velocity. In MoSi2 single crystals of a soft orientation, dislocations with 1/2<111> Burgers vectors are created in localized sources and move on {110} planes in a viscous manner. The dislocations in Al-Pd-Mn single quasicrystals are oriented in preferred crystallographic directions and move in a viscous way as well. On the basis of in situ observations, conclusions are drawn for interpreting macroscopic deformation behavior at high temperatures.

Journal Article↗

Modulating factors of calcium-free contraction at low [MgATP]: a physiological study on the steady states of skinned fibres of frog skeletal muscle.

Factors that modulate Ca(2+)-free contraction at low [MgATP] were examined by analysing steady tension development in skinned fibres of frog skeletal muscle. The commonly accepted bell-shaped relationship between steady tension and log (1/[MgATP]) was found to be highly susceptible to subtle experimental conditions at the higher [MgATP] side (right limb). The limb shifted to the right with increased fibre thickness, interrupted stirring of the bathing solution, increased temperature and fibre extension, although the effects of temperature and extension were marked only in thick fibres (cross-sectional area > 6000 microns 2). The shift of the right limb was reproduced by an addition of ADP to the bathing solution. These results, together with the extreme steepness of the right limb in thick fibres, suggest that a diffusion-dependent self-regenerative activation occurs in thick fibres in which ADP accumulation and ATP depletion positively feed back through further activation of the myofibrillar ATPase. Numerical simulation supported the hypothesis of the self-regenerative activation under poor diffusion conditions, and suggested that a small rise in temperature and fibre extension can trigger the self-regenerative process at the right limb. Consequently, ADP, temperature and fibre extension are deduced to be the primary potentiators of the activation at low [MgATP]. The high efficiency of ADP in shifting the limb suggests that the activating efficiency of the MgADP-bound actomyosin complex is higher than the nucleotide-free actomyosin complex.

Actomyosin↗

Expression of calcium-binding protein regucalcin mRNA in fetal rat liver is stimulated by calcium administration.

The expression of hepatic calcium-binding protein regucalcin mRNA in fetal rats was investigated. The alteration in regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.9 kb with complete open reading frame). Hepatic regucalcin mRNA levels were progressively increased with fetal development; the mRNA was clearly expressed at 15 and 21 days of pregnancy but only slightly at the 8 days. Meanwhile, beta-actin mRNA levels in the fetal liver were remarkable at 8 and 15 days of pregnancy. The fetal liver regucalcin mRNA levels at 15 days of pregnancy were significantly decreased by overnight-fasting of maternal rats. The oral administration of calcium chloride (50 mg Ca/100 g body weight) to maternal rats at 15 days of pregnancy caused a remarkable elevation (about 2 fold) of regucalcin mRNA levels in the fetal liver; this increase was seen 60 and 180 min after the calcium administration. After birth, regucalcin mRNA was increasingly expressed in the livers of newborn and weanling rats, while hepatic beta-actin mRNA expression was not appreciably altered with increasing ages. These findings demonstrate that the expression of hepatic regucalcin mRNA is increased with fetal development, and that the gene expression may be stimulated by the ingestion of dietary calcium.

Animals↗

Alteration in calcium content and Ca2+-ATPase activity in the liver nuclei of rats orally administered carbon tetrachloride.

The alteration in calcium transport in the liver nuclei of rats orally administered carbon tetrachloride (CCl4) was investigated. Rats received a single oral administration of CCl4 (5, 10, and 25%, 1.0 ml/100 g body weight), and 5, 24 and 48 h later the animals were sacrificed. The administration of CCl4 (25%) caused a remarkable elevation of calcium content in the liver tissues and the nuclei of rats. Liver nuclear Ca2+-ATPase activity was markedly decreased by CCl4 (25%) administration. The presence of dibutyryl cyclic AMP(10(-4) and 10(-3) M) or inositol 1,4,5-trisphosphate (10(-6) and 10(-5) M) in the enzyme reaction mixture caused a significant decrease in Ca2+-ATPase activity in the liver nuclei obtained from normal rat, while the enzyme activity was significantly increased by calmodulin (1.0 and 2.0 microg/ml). These signaling factor's effects were completely impaired in the liver nuclei obtained from CCl4 (25%)-administered rats. DNA fragmentation in the liver nuclei obtained from CCl4-administered rats was significantly decreased by the presence of EGTA (2 mM) in the reaction mixture, suggesting that the endogenous calcium activates nuclear DNA fragmentation. The present study demonstrates that calcium transport system in the liver nuclei is impaired by liver injury with CCl4 administration in rats.

Administration, Oral↗

Anabolic effect of genistein and genistin on bone metabolism in the femoral-metaphyseal tissues of elderly rats: the genistein effect is enhanced by zinc.

The effect of genistein and genistin on bone components in the femoral-metaphyseal tissues obtained from elderly female rats was investigated in vitro. The metaphyseal tissues were cultured for 24 h in a medium containing either vehicle, genistein (10(-8)-10(-5) M) or genistin (10(-7)-10(-5) M). The presence of genistein or genistin caused a significant increase in alkaline phosphatase activity, deoxyribonucleic acid (DNA) and calcium contents in the metaphyseal tissues. The effect of genistein was greater than that of genistin. The bone components increased by genistein (10(-5) M) or genistin (10(-5) M) were completely blocked by the presence of cycloheximide (10(-6) M). The presence of zinc sulfate (10(-5) M) caused a significant increase in the genistein (10(-5) M)-elevated alkaline phosphatase activity, DNA and calcium contents. The enhancement with zinc was not seen by genistin (10(-5) M). The stimulatory effect of zinc on the genistein-induced increase in bone components of the metaphyseal tissues was completely blocked by the presence of cycloheximide (10(-6) M). The present results suggest that genistein and genistin have an anabolic effect on bone metabolism in the femoral-metaphyseal tissues of elderly rats, and that the genistein effect is enhanced by zinc, an essential trace element.

Aging↗

Tissue-specific binding of nuclear factors to the 5'-flanking region of the rat gene for calcium-binding protein regucalcin.

The existence of nuclear factors which bind to the 5'-flanking region of calcium-binding protein regucalcin gene in rats was investigated. We previously reported that rat regucalcin mRNA is expressed in a highly tissue-specific manner; the mRNA was mainly present in the liver but only slightly in the kidney. When the nuclear proteins extracted from the liver and kidney of rats were used in the gel mobility shift assays, a protein-DNA complex was uniquely formed with the DNA fragment containing the upstream region from the first exon of rat regucalcin gene. On the other hand, this complex was not found by using the nuclear extracts from rat brain, spleen, and heart. The nuclear proteins of these extracts, however, could specifically bind to the DNA fragment containing the first exon region of rat regucalcin gene, although Northern blot analysis did not show detectable amount of regucalcin mRNA levels in rat brain, spleen, and heart. The present study demonstrates that the existence of nuclear protein components which bind to the regucalcin gene. These identified components may be involved in the tissue-specific regulation of regucalcin gene expression.

Animals↗

Streptozotocin-induced diabetes increases (Ca2+-Mg2+)-ATPase activity in hepatic plasma membranes of rats: involvement of protein kinase C.

The alteration in calcium transport in the liver of rats with streptozocin(STZ)-diabetic state was investigated. STZ (6 mg/100 g body weight) was subcutaneously administered in rats, and 1 or 2 weeks later they were sacrificed by bleeding. STZ administration caused a remarkable elevation of serum glucose concentration. Liver calcium content was significantly increased by STZ administration. Hepatic plasma membrane (Ca2+-Mg2+)-ATPase activity was markedly elevated by STZ administration. This increase was completely abolished by the presence of staurosporine (10(-7)-10(-5) M), an inhibitor of protein kinase C, in the enzyme reaction mixture, suggesting an involvement of protein kinase C signalling. Moreover, the STZ-induced increase in liver plasma membrane (Ca2+-Mg2+)-ATPase activity was significantly raised by the presence of okadaic acid (10(-5) and 10(-4) M). Meanwhile, the STZ-increased (Ca2+-Mg2+)-ATPase activity was not appreciably altered by the presence of anti-regucalcin IgG in the reaction mixture, indicating that the activatory protein regucalcin does not participate in the elevation of the enzyme activity. The present study demonstrates that STZ-induced diabetes causes the increase in hepatic plasma membrane (Ca2+-Mg2+)-ATPase activity of rats.

Animals↗

Stimulatory effect of calcium administration on regucalcin mRNA expression is attenuated in the kidney cortex of rats ingested with saline.

The expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats ingested with saline was investigated. The alteration in regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb of open reading frame). Rats were freely given saline as drinking water for 7 days. Regucalcin mRNA levels in the kidney cortex were suppressed by saline ingestion. When calcium chloride (10 mg Ca/100 g body weight) was intraperitoneally administered to rats ingested with saline for 7 days, the effect of calcium administration to increase regucalcin mRNA levels was weakened by saline ingestion. Such effect was also seen by the administration of 2.5 and 5 mg Ca/100 g. Regucalcin mRNA levels in the kidney cortex of spontaneous hypertensive rats (SHR) were not appreciably increased by the administration of calcium (10 mg/100 g). Meanwhile, calcium content in the kidney cortex was significantly elevated by the administration of calcium (10 mg/100 g) to normal rats. This increase was weakened in saline-ingested rats. Moreover, Ca2+/calmodulin-dependent protein kinase activity in the cytosol of kidney cortex was significantly decreased by saline ingestion. These results suggest the possibility that saline ingestion-induced suppression of regucalcin mRNA expression in the kidney cortex is partly involved in the attenuation of Ca2+ signalling.

Animals↗

Descriptive epidemiology of body mass index in Japanese adults in a representative sample from the National Nutrition Survey 1990-1994.

OBJECTIVE: To determine the prevalence of obesity and reference percentile curves of body mass index (BMI; kg/m2) in Japanese adults. DESIGN: Cross-sectional nationwide surveys (The National Nutrition Survey, Japan) carried out in 1990-1994. A sample of 23,556 males and 28,751 females aged 15-84 y was used for this analysis. RESULTS: The prevalence of grades 1 (BMI: 25.0-29.9), 2 (30.0-39.9) and 3 (> or = 40.0) overweight, was estimated to be 24.3%, 1.84% and 0.019% in males, respectively, and 20.2%, 2.87% and 0.032% in females, respectively, subjects aged 35-64 y after standardization to the age-distribution of the standard world population (WHO, 1993). The age-standardized prevalence of obesity (for BMI > or = 26.4, a criterion of the Japan Society for the Study of Obesity; and > or = 30.0 was 13.6% and 1.86% in males, respectively, and 13.5% and 2.90% in females, respectively, subjects aged 35-64 y. CONCLUSIONS: The standardized prevalence of obesity (BMI > or = 30.0) in Japanese adults was quite low compared with the data in western populations.

Adolescent↗

Biphasic expression of CD4 in acute myelocytic leukemia (AML) cells: AML of monocyte origin and hematopoietic precursor cell origin.

In 227 of 495 (45.9%) Japanese adult patients with acute myelocytic leukemia (AML), leukemic cells expressed CD4. Incidence of CD4 expression in each FAB subtype was as follows: M1 37.4%, M2 33.7%, M3 35.4%, M4 65.0%, and M5 78.3%. The typical expression pattern of myelomonocytic differentiation antigens and cytokine receptors in CD4+ AML was CD34lowCD33high CD11bhighGM-CSFRhigh. AML cases with 11q23 abnormalities and with inv(16) were frequently CD4-positive. These data collectively indicate that CD4 expression in AML cells is associated with monocytic characteristics. However, CD4+CD34high AML cases appear to have unique immature characteristics including low expression of myelomonocytic differentiation antigens (ie CD33 and CD11b), and accumulation of chromosome abnormalities (ie t(8;21) in CD4lowCD34high AML and chromosome 7 abnormalities in CD4highCD34high AML). We speculate that these leukemia subsets originate from CD4+ hematopoietic precursor cells, therefore then should be considered separately from most of the CD4+ AML as represented by CD34lowCD33high CD11bhighGM-CSFRhigh. Overall survival of patients with CD4+ AML in our series was worse than that of those with CD4 AML (P = 0.0202).

Adult↗

Culture system for extensive production of CD19+IgM+ cells by human cord blood CD34+ progenitors.

We established a co-culture system with a monolayer of the murine bone marrow (BM) stroma cell line, MS-5, in which human cord blood CD34+ cells differentiated to CD19+ cells. The addition of stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF) highly enhanced the production of CD19+ cells. The expansion of the cell numbers was over 10(3)-fold. Furthermore, a significant proportion (<45%) of the cells expressed surface IgM (sIgM) after 5 weeks of co-culture. CD34+CD19- cells also showed a similar development of CD19+ cells and CD19+sigM+ cells. Filter separation of MS-5 cells and CD34+ cells did not inhibit the growth of CD19+ cells. However, when further purified CD34+CD19-CD13- CD33- cells were cultured in the presence of MS-5 cells with or without a separation filter, CD19+ cells did not appear in the non-contact setting. This result suggested that the highly purified CD34+CD19-CD13-CD33- progenitors require the cell-cell contact for the development of CD19+ cells, whereas other CD34+ fractions contain progenitors that do not require the contact. This co-culture system should be useful for the study of early human B-lymphopoiesis.

Animals↗

Simultaneous determination of urinary catecholamines and 5-hydroxyindoleamines by high-performance liquid chromatography with fluorescence detection.

A simple, selective and sensitive high-performance liquid chromatographic method with fluorescence detection is described for the simultaneous determination of catecholamines and 5-hydroxyindoleamines. The method is based on the derivatization of the amines with benzylamine in the presence of potassium hexacyanoferrate(III) under mild conditions. The resulting fluorescent derivatives are separated on a reversed-phase column, Cosmosil 5C18, using a mixture of acetonitrile and 10 mmol l-1 acetate buffer (pH 6.0) (35 + 65 v/v) as mobile phase, and are detected spectrofluorimetrically at 480 nm with excitation at 345 nm. The detection limits (signal-to-noise ratio = 3) for the amines are 2.8-236.0 fmol for an injection volume of 50 microliters. The method was successfully applied to the determination of 5-hydroxyindol-3-ylacetic acid, serotonin, norepinephrine and epinephrine in human urine. The method is simple and does not require clean-up of urine samples.

Benzylamines↗

Population structure of nuclear and mitochondrial DNA variation among humpback whales in the North Pacific.

The population structure of variation in a nuclear actin intron and the control region of mitochondrial DNA is described for humpback whales from eight regions in the North Pacific Ocean: central California, Baja Peninsula, nearshore Mexico (Bahia Banderas), offshore Mexico (Socorro Island), southeastern Alaska, central Alaska (Prince Williams Sound), Hawaii and Japan (Ogasawara Islands). Primary mtDNA haplotypes and intron alleles were identified using selected restriction fragment length polymorphisms of target sequences amplified by the polymerase chain reaction (PCR-RFLP). There was little evidence of heterogeneity in the frequencies of mtDNA haplotypes or actin intron alleles due to the year or sex composition of the sample. However, frequencies of four mtDNA haplotypes showed marked regional differences in their distributions (phi ST = 0.277; P < 0.001; n = 205 individuals) while the two alleles showed significant, but less marked, regional differences (phi ST = 0.033; P < 0.013; n = 400 chromosomes). An hierarchical analysis of variance in frequencies of haplotypes and alleles supported the grouping of six regions into a central and eastern stock with further partitioning of variance among regions within stocks for haplotypes but not for alleles. Based on available genetic and demographic evidence, the southeastern Alaska and central California feeding grounds were selected for additional analyses of nuclear differentiation using allelic variation at four microsatellite loci. All four loci showed significant differences in allele frequencies (overall FST = 0.043; P < 0.001; average n = 139 chromosomes per locus), indicating at least partial reproductive isolation between the two regions as well as the segregation of mtDNA lineages. Although the two feeding grounds were not panmictic for nuclear or mitochondrial loci, estimates of long-term migration rates suggested that male-mediated gene flow was several-fold greater than female gene flow. These results include and extend the range and sample size of previously published work, providing additional evidence for the significance of genetic management units within oceanic populations of humpback whales.

Alleles↗

cDNA cloning, gene expression and subcellular localization of anthocyanin 5-aromatic acyltransferase from Gentiana triflora.

Acylation of anthocyanins with hydroxycinnamic acid derivatives is one of the most important and less under-stood modification reactions during anthocyanin biosynthesis. Anthocyanin aromatic acyltransferase catalyses the transfer of hydroxycinnamic acid moieties from their CoA esters to the glycosyl groups of anthocyanins. A full-length cDNA encoding the anthocyanin 5-aromatic acyltransferase (5AT) (EC 2.3.1.153) that acylates the glucose bound at the 5-position of anthocyanidin 3,5-diglucoside was isolated from petals of Gentiana triflora on the basis of the amino acid sequence of the purified enzyme. The isolated full-length cDNA had an open reading frame of 469 amino acids and the calculated molecular weight was 52,736. The deduced amino acid sequence contains consensus motifs that are conserved among the putative acyl CoA-mediated acyltransferases, and this indicates that 5AT is a member of a proposed superfamily of multi-functional acyltransferases (St-Pierre et al. (1998) Plant J. 14, 703-713). The cDNA was expressed in Escherichia coli and yeast, and confirmed to encode 5AT. The enzymatic characteristics of the recombinant 5AT were consistent with those of the native gentian 5AT. Immunoblot analysis using specific antibodies to 5AT showed that the 5AT protein is present in petals, but not in sepals, stems or leaves of G. triflora. RNA blot analysis showed that the 5AT gene is expressed only in petals and that its expression is temporally regulated during flower development coordinately with other anthocyanin biosynthetic genes. Immunohistochemical analysis demonstrated that the 5AT protein is specifically expressed in the outer epidermal cells of gentian petals and that it is localized mainly in the cytosol.

Acyltransferases↗

A rice homolog of Cdk7/MO15 phosphorylates both cyclin-dependent protein kinases and the carboxy-terminal domain of RNA polymerase II.

The activation of cyclin-dependent protein kinases (CDKs) requires phosphorylation of a threonine residue within the T-loop by a CDK-activating kinase (CAK). The R2 protein of rice is very similar to CAKs of animals and fission yeast at the amino acid level but phosphorylation by R2 has not yet been demonstrated. When R2 was overexpressed in a CAK-deficient mutant of budding yeast, it suppressed the temperature sensitivity of the mutation. Immunoprecipitates of rice proteins with the anti-R2 antibody phosphorylated human CDK2, one of the rice CDKs (Cdc2Os1), and the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II of Arabidopsis. Mutational analysis indicated that R2 phosphorylated the threonine residue within the T-loop of CDK2 and Cdc2Os1. R2 was found mainly in two protein complexes which had molecular masses of 190 kDa and 70 kDa, respectively, whilst the CDK- and CTD-kinase activities associated with R2 were identified in a complex of 105 kDa. These results indicate that R2 is closely related to CAKs of animals and fission yeast in terms of its phosphorylation activity and, moreover, that this CAK of rice is distinct from a CAK of the dicotyledonous plant Arabidopsis.

Animals↗

Efficacy of preconditioning with N-acetylcysteine against reperfusion injury after prolonged cold ischaemia in rats liver in which glutathione had been reduced by buthionine sulphoximine.

OBJECTIVE: To investigate the ability of N-acetylcysteine (NAC) to prevent cold ischaemic-reperfusion injury and improve hepatic integrity in a glutathione-depleted condition. DESIGN: Open laboratory study. SETTING: University hospitals, Japan and France. MATERIALS: 40 male Wistar rats. INTERVENTIONS: To produce a glutathione-depleted liver, buthionine sulphoximine (BSO) was injected intraperitoneally 2 hours before either NAC or 5% dextrose was infused 15 minutes before the liver was harvested. We used an isolated perfused rat liver model that had undergone prolonged hypothermic ischaemia, cold-storage for 48 hours and reperfusion for 120 minutes. MAIN OUTCOME MEASURES: Concentrations of hepatic enzymes released into samples of perfusate, concentration of adenosine triphosphate in liver tissue, concentrations of reduced and oxidized glutathione in perfusate, and bile production. RESULTS: The concentrations of the hepatocellular enzymes and oxidised glutathione in the perfusate samples were significantly reduced in the NAC group compared with the 5% dextrose group. Bile production improved significantly in the NAC group compared with the 5% dextrose group. The concentration of reduced glutathione in liver tissue was not increased by NAC. CONCLUSION: In a glutathione-depleted liver NAC prevented hepatic injury and improved liver integrity after a cold ischaemic-reperfusion injury, by acting not as a substrate for glutathione synthesis but as a direct free radical scavenger.

Acetylcysteine↗

Pharmacokinetics of soybean isoflavones in plasma, urine and feces of men after ingestion of 60 g baked soybean powder (kinako).

To take advantage of the various pharmacologic activities of soy bean isoflavones, more detailed studies of the absorption and excretion rates of these compounds in humans and subsequent evaluation of their bioavailabilities are required. We conducted a pharmacokinetic study of soybean isoflavones in seven healthy male volunteers. After ingestion of 60 g of kinako (baked soybean powder, containing 103 micromol daidzein and 112 micromol genistein), changes of the isoflavone and metabolite concentrations in plasma, urine and feces were measured by gas chromatography-mass spectrometry. The plasma concentration of genistein increased after 2 h and reached its highest value of 2.44 +/- 0.65 micromol/L 6 h later. The plasma concentration of daidzein peaked at 1.56 +/- 0.34 micromol/L at the same time, but it was always lower than that of genistein. Peak plasma concentration of O-desmethylangolensin (O-DMA) and equol appeared after the daidzein peak in four and two subjects, respectively. In contrast with plasma, daidzein was the main component in urine. Urinary daidzein excretion started to increase shortly after the rise in its plasma concentration and reached 2.4 micromol/h 8 h after ingestion of kinako. Genistein excretion in urine paralleled that of daidzein, but the value at 6 h was about half (1.1 micromol/h). The majority of ingested isoflavones after ingestion of kinako were recovered on d 2 or 3 in the feces. Total recovery of daidzein, O-DMA and equol from urine and feces was 54.7%, calculated from daidzein intake; 20.1% of administered genistein was recovered as genistein. The half-lives of plasma genistein and daidzein were 8.36 and 5.79 h, respectively. The individual plasma and urinary concentrations of equol and O-DMA were quite variable; subjects were classified as high and low metabolizers. The high plasma concentration of isoflavones for at least several hours after a single ingestion of soy protein suggests that these compounds may interact with macromolecules and have biological effects.

Absorption↗