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Biomedical subjects

M Yokoi

Publications and source records attributed to M Yokoi.

At least 37 records · Page 2Linked to original sources

Favorable life-style modification and attenuation of cardiovascular risk factors.

In order to develop an effective counseling system for prevention of cardiovascular diseases, the association of a favorably changed life-style with improved risk factors was examined. Participants were 7,321 office workers aged 30-69 years from in and around Nagoya city. The age-adjusted odds ratio (OR) and its 95% confidence interval (CI) were calculated to assess the likelihood of risk factor improvement by favorable life-style modifications during a 3-year period. Those who began to eat breakfast and increased their vegetable intake normalized their previously abnormal diastolic blood pressure with more than twice the likelihood (adjusted OR [95% CI] 2.89 [1.29-6.46] and 2.60 [1.18-5.75], respectively). 'Began to eat breakfast' was also significantly associated with normalized total cholesterol (TC) (1.84, [1.05-3.21]). 'Stopped eating till full' significantly normalized the body mass index (2.03; [1.25-3.28]), uric acid (1.65; [1.07-2.52]) and TC (1.43; [1.04-1.97]). Those who started regular exercise significantly normalized their high-density lipoprotein-cholesterol (HDL-C) abnormality with 1.69-times the likelihood (1.69; [1.24-2.29]) and those who began to walk briskly also improved their TC abnormality (1.85; [1.19-2.89]). HDL-C was normalized with 2.55-times the likelihood in those who quit smoking (2.55; [1.68-3.86]). Because favorable life-style modifications can attenuate abnormal cardiovascular risk factors, then proper advice on specific risk factors should be routinely given at each health check-up in order to prevent the onset of cardiovascular diseases in subsequent years.

Adult↗

Preparation and application of anti-idiotypic antibody against anti-gibberellin A4 antibody.

A monoclonal anti-idiotypic antibody was raised against anti-gibberellin A4 (GA4) antibody, which recognizes biologically active gibberellins such as GA1 and GA4 specifically. Amino acid sequences of variable regions of both anti-GA4 and anti-idiotypic antibodies were analyzed. By using the property of the anti-idiotypic antibody to compete with GA1/4 in binding to the anti-GA4 antibody, we successfully applied the anti-idiotypic antibody to ELISA as a tracer for measuring GA1/4. The single-chain Fv (scFv) gene of the anti-idiotypic antibody was constructed, and scFv expressed in E. coli showed binding activity to anti-GA4 antibody. These results suggest the possible application of anti-idiotypic antibody as a handy and stable source of an enzymatic tracer for ELISA by production of fusion protein of the scFv and an appropriate enzyme.

Amino Acid Sequence↗

Ablation of cerebellar Golgi cells disrupts synaptic integration involving GABA inhibition and NMDA receptor activation in motor coordination.

The role of inhibitory Golgi cells in cerebellar function was investigated by selectively ablating Golgi cells expressing human interleukin-2 receptor alpha subunit in transgenic mice, using the immunotoxin-mediated cell targeting technique. Golgi cell disruption caused severe acute motor disorders. These mice showed gradual recovery but retained a continuing inability to perform compound movements. Optical and electrical recordings combined with immunocytological analysis indicated that elimination of Golgi cells not only reduces GABA-mediated inhibition but also attenuates functional NMDA receptors in granule cells. These results demonstrate that synaptic integration involving both GABA inhibition and NMDA receptor activation is essential for compound motor coordination. Furthermore, this integration can adapt after Golgi cell elimination so as not to evoke overexcitation by the reduction of NMDA receptors.

Animals↗

Observation of composition in surface monolayers by X-ray scattering spectra caused by crystal truncation and interferences.

X-ray crystal-truncation-rod (CTR) scattering measurements using synchrotron radiation and an imaging plate can reveal the composition of a surface monolayer. Even when the composition is changed in only one atomic layer on the top surface, the X-ray CTR spectrum can change due to the differences in composition. X-ray CTR spectra are greatly enhanced by X-ray interference when a sample is designed properly. In this paper, it is shown by theoretical calculations and experiments for AlAs/GaAs samples grown by MBE that a 1 ML (monolayer)-thick AlAs layer embedded under 10 ML below the surface can enhance the modulation of an X-ray CTR spectrum.

Journal Article↗

The second-largest subunit of the mouse DNA polymerase alpha-primase complex facilitates both production and nuclear translocation of the catalytic subunit of DNA polymerase alpha.

DNA polymerase alpha-primase is a replication enzyme necessary for DNA replication in all eukaryotes examined so far. Mouse DNA polymerase alpha is made up of four subunits, the largest of which is the catalytic subunit with a molecular mass of 180 kDa (p180). This subunit exists as a tight complex with the second-largest subunit (p68), whose physiological role has remained unclear up until now. We set out to characterize these subunits individually or in combination by using a cDNA expression system in cultured mammalian cells. Coexpression of p68 markedly increased the protein level of p180, with the result that ectopically generated DNA polymerase activity was dramatically increased. Immunofluorescence analysis showed that while either singly expressed p180 or p68 was localized in the cytoplasm, cotransfection of both subunits resulted in colocalization in the nucleus. We identified a putative nuclear localization signal for p180 (residues 1419 to 1437) and found that interaction with p68 is essential for p180 to translocate into the nucleus. These results indicate that association of p180 with p68 is important for both protein synthesis of p180 and translocation into the nucleus, implying that p68 plays a pivotal role in the newly synthesized DNA polymerase alpha complex.

3T3 Cells↗

Newly developed diabetic retinopathy and its preceding changes in biological markers.

To disclose the chronological changes prior to the manifestation of diabetic retinopathy (DR), we analyzed the time course of biological markers among apparently healthy diabetic subjects in a case-control study of 21,579 adults who had undergone comprehensive health examinations for > or = 10 years. We identified 54 cases who had newly developed DR, and selected 108 adults without fundus abnormalities, matching them for sex, age, and fasting plasma glucose (FPG) at the onset of the patient group's retinopathy as a referent group from the same population. In a multivariate analysis, a high average FPG (> 175 mg/dl) and a final-year FPG reduction (< -3%) were significantly associated with a 5.4 (95% CI, 1.8-15.7)- and 5.0 (95% CI, 1.0-24.7)-fold increased risk of DR, respectively. Thus, we surmised that sustained hyperglycemia and a subsequent drop in FPG might promote retinopathy in non-insulin dependent diabetes mellitus.

Biomarkers↗

Relation of electrocardiographic left ventricular hypertrophy with and without T-wave changes to systemic blood pressure, body mass, and serum lipids and blood glucose levels in Japanese men.

Left ventricular (LV) hypertrophy, especially combined with an abnormal ST-T, is considered 1 of many coronary risk factors. Seven hundred forty-nine Japanese men were selected according to their electrocardiographic findings, i.e., normal electrocardiogram, LV hypertrophy without an abnormal ST-T segment, LV hypertrophy with a flat T wave, and LV hypertrophy with a negative T wave. Coronary risk factors were compared among these 4 age-matched groups. Groups with LV hypertrophy with negative or flat T waves had larger body mass index (24.9 vs 22.9 kg/m2), higher mean systemic blood pressure (111 vs 95 mm Hg), larger LV mass (265 vs 157 g), higher blood glucose (110 vs 100 mg/dl), higher serum triglyceride (148 vs 122 mg/dl), higher total cholesterol (206 vs 198 mg/dl), and lower high-density lipoprotein cholesterol (47 vs 54 mg/dl) than the normal group or the group with LV hypertrophy without T-wave change. Among these risk factors, blood pressure and glucose remained higher even after the adjustment by body mass index or by body mass index and blood pressure. Electrocardiographic LV hypertrophy with a changed T wave signified higher risk of coronary artery disease in Japanese men.

Blood Glucose↗

Molecular cloning of the cDNA for the catalytic subunit of plant DNA polymerase alpha and its cell-cycle dependent expression.

BACKGROUND: DNA polymerase alpha has been studied in considerable detail in yeast and animals. Genetic and biochemical analyses reveal that this enzyme is composed of a heterotetramer and is necessary for replicon initiation and primer synthesis in lagging strand synthesis. In spite of the fact that modes of DNA replication in plants seem to be similar to those in other eukaryotes, very little is known about the biochemical components that participate in DNA replication of plants, including DNA polymerases. RESULTS: Using a 561-base pair DNA fragment, obtained by polymerase chain reaction amplification from a rice cDNA library as a probe, we isolated and sequenced a cDNA homologous to the cDNA for the catalytic subunit of rice DNA polymerase alpha. The encoded polypeptide has extensive homology with the catalytic subunit of DNA polymerase alpha from several species. Furthermore, when the cDNA was expressed in eukaryotic transcription/translation systems, the protein products showed DNA polymerase activity which was inhibited by a monoclonal antibody specific for DNA polymerase alpha. Using RNA gel blot analysis, we found that the levels of mRNA of the catalytic subunit of this enzyme is regulated during the cell-cycle in plant cells. CONCLUSION: This is the first report which describes the cDNA cloning of plant DNA polymerase. We conclude that the principal features of the DNA polymerase alpha catalytic subunit are conserved in plants.

Amino Acid Sequence↗

Impairment of hippocampal mossy fiber LTD in mice lacking mGluR2.

Subtype 2 of the metabotropic glutamate receptor (mGluR2) is expressed in the presynaptic elements of hippocampal mossy fiber-CA3 synapses. Knockout mice deficient in mGluR2 showed no histological changes and no alterations in basal synaptic transmission, paired-pulse facilitation, or tetanus-induced long-term potentiation (LTP) at the mossy fiber-CA3 synapses. Long-term depression (LTD) induced by low-frequency stimulation, however, was almost fully abolished. The mutant mice performed normally in water maze learning tasks. Thus, the presynaptic mGluR2 is essential for inducing LTD at the mossy fiber-CA3 synapses, but this hippocampal LTD does not seem to be required for spatial learning.

Animals↗

Acylated pelargonidin 3-sambubioside-5-glucosides in Matthiola incana.

Ten acylated pelargonidin 3-sambubioside-5-glucosides were isolated from the red-purple flowers of Matthiola incana, and also pelargonidin 3-glucoside was isolated from the brownish-red flowers of this plant. FAB mass measurements of 10 acylated anthocyanins gave their molecular ions [M]+ at 903-1195 m/z, which were based on acylated pelargonidin 3-sambubioside-5-glucosides with malonic acid, sinapic acid, p-coumaric acid, caffeic acid and/or ferulic acid. This was confirmed by the analysis of NMR spectra and the experiments of acid and alkaline hydrolysis. By spectral and chemical methods, seven of the 10 pigments were determined to be pelargonidin 3-O-[2-O-(2-O-(acyl-I)-beta-D-xylopyranosyl)- 6-O-(acyl-II)-beta-D-glucopyranoside]-5-O-[6-O-(malonyl)-beta-D- glucopyranoside], in which acyl moieties varied between sinapic, ferulic, caffeic and p-coumaric acids. The occurrence of these pigments was examined in 10 red-purple, 10 salmon-pink, three apricot and three copper colour cultivars of M. incana by HPLC. The acylated pelargonidin 3-sambubioside-5-glucosides were present as the dominant pigments in the red-purple, salmon-pink and apricot colour cultivars. On the other hand, pelargonidin 3-glucoside was present as a dominant anthocyanin in the copper colour cultivars and also pelargonidin 3-sambubioside-5-glucoside was confirmed by HPLC as a minor pigment in the copper colour flowers.

Acylation↗

Acylated pelargonidin glycosides in red-purple flowers of Ipomoea purpurea.

Four acylated pelargonidin glycosides were isolated from the red-purple flowers of Ipomoea purpurea. The acylated anthocyanins were all based on pelargonidin 3-sophoroside-5-glucoside, acylated with caffeic acid and/or glucosylcaffeic acids. Three novel anthocyanins were elucidated to be pelargonidin 3-O-[2-O-(6-O-(trans-3-O-(beta-D-glucopyranosyl)-caffeyl)- beta-D-glucopyranosyl)-6-O-(trans-4-O-(6-O-(trans-caffeyl)-beta-D- glucopyranosyl)-caffeyl)-beta-D-glucopyranoside]-5-O- [beta-D-glucopyranoside], pelargonidin 3-O-[2-O-(6-O-(trans-caffeyl)-beta-D-glucopyranosyl)-6-O- (trans-4-O-(6-O-(trans-caffeyl)-beta-D-glucospyranosyl)-ceffeyl )-beta-D- glucopyranoside]-5-O-[beta-D-glucopyranoside] and pelargonidin 3-O[2-O-(2-O-(6-O-(trans-caffeyl)-beta-D-glucopyranosyl)-6-O- (trans-caffeyl)-beta-D-glucopyranoside]-5-O-[beta-D-glucopyoside]. Another anthocyanin was pelargonidin 3-O-[2-O-(glucosylcaffeylglucosyl)-6-O-(caffeyl)-glucoside]-5-gluc oside, which was reported to be present in the red-purple flowers of Pharbitis nil. Apart from the second pigment cited, these are analogous pigments of the violet-blue Ipomoea anthocyanins which are composed of acylated cyanidin glycosides, instead of pelargonidin.

Acylation↗

Identification of the nuclear localization signal of mouse DNA primase: nuclear transport of p46 subunit is facilitated by interaction with p54 subunit.

DNA polymerase alpha-primase is a replication enzyme necessary for DNA replication in all eukaryotes. Mouse DNA primase is composed of two subunits: a 46 kDa protein (p46), which is the catalytic subunit capable of RNA primer synthesis, and a 54 kDa protein (p54), whose physiological role is not clear. To understand the structure-function relationship of DNA primase, we set out to characterize these two subunits individually or in combination using a cDNA expression system in mammalian cultured cells, and determined the subcellular distribution of ectopically expressed DNA primase. The p54 expressed in COS-1 cells after transfection was predominantly localized in the nucleus, whereas p46 was retained in the cytoplasm as shown by indirect immunofluorescence analysis. Using several mutant proteins with deletions or substitutions as well as chimeric constructs, we identified the nuclear localization signal of p54 as RIRKKLR, encoded near the amino terminus (residues 6-12). Furthermore, co-expression of both p46 and p54 subunits markedly altered the subcellular distribution of p46; co-expressed p46 was transported into the nucleus as efficiently as p54. These results demonstrate that p54 has a nuclear localization signal and is able to be translocated into the nucleus independently of DNA polymerase alpha subunits. In contrast, p46 lacks a nuclear localization signal, and its nuclear translocation is facilitated by interaction with p54. We present here first evidence for a novel role of p54 in the nuclear translocation process, and a piggy-back binding transport mechanism of mouse DNA primase.

3T3 Cells↗

Refinement of odor molecule tuning by dendrodendritic synaptic inhibition in the olfactory bulb.

Mitral/tufted cells (M/T cells) and granule cells form reciprocal dendrodendritic synapses in the main olfactory bulb; the granule cell is excited by glutamate from the M/T cell and in turn inhibits M/T cells by gamma-aminobutyrate. The trans-synaptically excited granule cell is thought to induce lateral inhibition in neighboring M/T cells and to refine olfactory information. It remains, however, elusive how significantly and specifically this synaptic regulation contributes to the discrimination of different olfactory stimuli. This investigation concerns the mechanism of olfactory discrimination by single unit recordings of responses to a series of normal aliphatic aldehydes from individual rabbit M/T cells. This analysis revealed that inhibitory responses are evoked in a M/T cell by a defined subset of odor molecules with structures closely related to the excitatory odor molecules. Furthermore, blockade of the reciprocal synaptic transmission by the glutamate receptor antagonist or the gamma-aminobutyrate receptor antagonist markedly suppressed the odor-evoked inhibition, indicating that the inhibitory responses are evoked by lateral inhibition via the reciprocal synaptic transmission. The synaptic regulation in the olfactory bulb thus greatly enhances the tuning specificity of odor responses and would contribute to discrimination of olfactory information.

Aldehydes↗

Acylated cyanidin 3-sambubioside-5-glucosides in Matthiola incana.

Four acylated cyanidin 3-sambubioside-5-glucosides were isolated from purple-violet flowers of Matthiola incana and their structures were determined by chemical and spectroscopic methods. Three acylated anthocyanins were cyanidin 3-O-(6-O-acyl-2-O-(2-O-sinapyl-beta-D-xylopyranosyl)-beta-D- glucopyranosides)-5-O-(6-O-malonyl-beta-D-glucopyranosides), in which the acyl group is p-coumaryl, caffeyl or ferulyl, respectively. The remaining pigment is free from malonic acid and was identified as cyanidin 3-O-(6-O-trans-ferulyl-2-O-(2- O-trans-sinapyl-beta-D-xylopyranosyl)-beta-D-glucopyranoside)-5-O- (beta-D-glucopyranoside). Analysis of the anthocyanin constituents in 16 purple-violet cultivars revealed that they contained the above triacylated anthocyanins in variable amounts as main pigments. An aromatic pair of pigments containing sinapic and ferulic acids are considered to produce an important intramolecular effect, making bluish colours in these flowers.

Acylation↗

Acylated cyanidin glycosides in the violet-blue flowers of Ipomoea purpurea.

Six acylated cyanidin glycosides were isolated from violet-blue flowers of Ipomoea purpurea. These anthocyanins were all based on cyanidin 3-sophoroside-5-glucoside, acylated with caffeic acid and/or p-coumaric acid. Three anthocyanin structures were elucidated to be cyanidin 3-O(-)[2-O-(6-O-(trans-3-O-(beta-D-glucopyranosyl)caffeyl)-beta -D-glucopyranosyl)-6-O-(trans-4-O-(6-O-(trans-caffeyl)-beta-D-glucopyran osyl) caffeyl)-beta-D-glucopyranoside]-5-O(-)[beta-D-glucopyranoside], cyanidin 3-O(-)[2-O-(trans-3-O-(beta-D-glucopyranosyl)caffeyl)-beta-D-glucopyrano syl)- 6-O-(trans-caffeyl)-beta-D-glucopyranoside]-5-O(-)[beta-D-glucopyranosid e d and cyanidin 3-O(-)[2-O-(6-O-(trans-caffeyl)-beta-D-glucopyranosyl)-6-O- (trans-caffeyl)-beta-D-glucopyranoside]-5-O(-)[beta-D-glucopyranoside]. These three anthocyanins were present in all 12 violet-blue flower strains as major pigments. The colours of these acylated antocyanins were stabler in neutral solution than their deacyl analogues.

Anthocyanins↗

[Intraoperative changes in blood lactate level and body temperature in younger and elderly adults].

Sequential changes in arterial blood lactate concentration and central-peripheral temperature gradient during anesthesia were measured in 10 younger patients (46-63 yr) and 10 elderly patients (66-92 yr) undergoing abdominal surgery. A central temperature probe was attached at the rectum and peripheral probe on the palm of the hand. Anesthesia was induced with thiopental and succinylcholine and maintained with isoflurane and nitrous oxide in oxygen. There was a marked increase in lactate values intraoperatively in the elderly as compared to the younger patients. Difference between central and peripheral temperature increased significantly in the elderly patients, while there was no change in the temperature gradient in the younger patients. Central-peripheral temperature gradient is an indicator of peripheral circulation, and the results imply the presence of anaerobic oxidation in elderly patients during operation. In elderly patients, the functional reserve of organ system and the tolerance to surgical stress are considered to be reduced. Intraoperative elevation of blood lactate levels might be caused either by impairment of peripheral circulation or by metabolic deficit due to age-related decline in hepatic function.

Abdomen↗