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Biomedical subjects

H Hayashi

Publications and source records attributed to H Hayashi.

At least 793 records · Page 44Linked to original sources

Spatial ventricular gradient in patients with Wolff-Parkinson-White syndrome in comparison with normal subjects: vectorcardiographic evidence for significant repolarization changes due to preexcitation.

We investigated the use of the spatial ventricular gradient (VG) from vectorcardiogram (VCG) to determine whether significant repolarization differences were present in patients with WPW syndrome compared with normal subjects and also examined which VG parameter (i.e., elevation, azimuth, and magnitude) reflected the differences in repolarization properties during preexcitation. VG was calculated in 49 patients of Wolff-Parkinson-White (WPW) syndrome (group A: left-sided accessory pathway, n = 29; group B: right-sided, n = 20). Group N consisted of 607 normal subjects. In group A, the azimuth of VG was significantly (p < 0.01) greater than in groups B and N. In group B, the elevation of VG was significantly (p < 0.01) greater than in groups A and N. There were no significant differences in the magnitude of VG among groups. QRS duration was significantly (p < 0.01) related with the elevation of VG in group B. These findings suggested that VG is useful for spatial evaluation of repolarization abnormalities during preexcitation, which are related to the site of the accessory pathway.

Adolescent↗

Purification and characterization of [3H]mepyramine (histamine H1 antagonist)-binding protein from rat liver: a highly homologous protein with cytochrome P450 2D.

A protein having a high-affinity binding site for [3H]mepyramine (MBP) was purified to homogeneity from rat liver membranes. The purified MBP has a single type of binding site for [3H]mepyramine with Kd value of 18.5 nM, and its molecular weight was determined to be 56,000 by SDS polyacrylamide gel electrophoresis. Amino acid sequences of twelve tryptic peptides derived from MBP are highly homologous with those of rat debrisoquine 4-hydroxylase (cytochrome P450 2D1) and other rat P450 2D subfamily members. In immunoblotting analysis, an antibody against rat P450 2D1 stained a band corresponding to MBP with Mr of 56,000; its migration position was clearly different from that of rat P450 2D1. Substrates and inhibitors of debrisoquine 4-hydroxylase potently displace [3H]-mepyramine binding to MBP. Quinine and quinidine showed 400 and 80 times, respectively, higher affinity for MBP than for debrisoquine 4-hydroxylase. These results suggest that MBP is a novel P450 2D family member.

Amino Acid Sequence↗

[New sex pheromone isolated from the abdominal glands of male newt, Cynops pyrrhogaster].

Sodefrin, a novel decapeptide which attracts female newts, Cynops pyrrhogaster was isolated from the abdominal gland of the male of the same species. Synthetic sodefrin was tested for its activity with sexually active females and males and sexually inert females. It attracted only sexually active females. The effect of sodefrin was blocked by a bilateral nostril plugging with cotton balls soaked in melted vaseline. Immunoelectronmicroscopic study using antiserum against sodefrin revealed that sodefrin was located mainly in the secretory granules in the epithelial cells of the abdominal gland of the cloaca.

Abdomen↗

Different responses of arterioles and venules in rat gastric mucosal microcirculation to endothelin-1 and endothelin-3.

Responses of the microvessels in the basal part of the gastric mucosa of anesthetized rats to endothelin (ET)-1 and ET-3 were examined by intravital microscopy. The posterior wall of the stomach, which was incised along the greater curvature, was secured in an observation chamber and superfused with Tyrode's solution, and the microcirculation was observed through a window made by removing a limited area of smooth-muscle layers with the serosa and submucosal tissue. Topical application of the same ranges of ET-1 and ET-3 (0.1-10 microM, 20 microliters) to the window dose-dependently constricted collecting venules and venules downstream. The constriction was inhibited by BQ-123, an ETA receptor antagonist, but the equipotency of ET-1 and ET-3 did not support the presence of the ETA receptor (tentatively called ETR-m). Arterioles were constricted through ETR-m, particularly at higher doses (> 3 microM), but 1 microM ET-1 dilated the arterioles by generating prostaglandin through BQ-123-sensitive ET receptor (tentatively called ETR-e), as shown by the inhibition of dilatation by indomethacin. Furthermore, the dilatation component of the arteriolar smooth-muscle responses to ET-1 and ET-3 was elicited by BQ-123 and was inhibited by NG-monomethyl-L-arginine (L-NMMA), indicating the generation of nitric oxide. This dilating effect of the endothelins was mediated by the ETB receptor.

Animals↗

Epidermal growth factor triggers the translocation of insulin-responsive glucose transporter (GLUT4).

In a novel cell line we developed for direct, sensitive detection of insulin-responsive glucose transporter (GLUT4) on the cell surface, we considered that insulin-activated phosphatidylinositol 3-kinase (PI 3-kinase) may be involved in the signaling pathway of insulin-stimulated GLUT4 translocation. We report here evidence that epidermal growth factor (EGF), which stimulates PI 3-kinase activity, also triggers GLUT4 translocation in Chinese hamster ovary (CHO) cells stably overexpressing the EGF receptor. The EGF-dependent GLUT4 translocation is possibly mediated by two independent pathways: one by PI 3-kinase and the other by protein kinase C (PKC); the PI 3-kinase-mediated pathway predominates. Triggering of the GLUT4 translocation is not specific for insulin, rather it may be a common property of growth factors which activate PI 3-kinase.

Androstadienes↗

[Thrombosed type aortic dissection showing changes in the false lumen to the patent type].

Thirty-four patients with acute aortic dissection with thrombosed false lumen were reviewed. In 4 cases (12%), the false lumens changed to the patent type. Changes in the false lumens were revealed by contrast-enhanced CT performed from day 2 to day 19 (mean, day 12.5) after the onset. The entry site was located at the descending aorta in 3 cases, and was unknown in 1 case. In 2 cases, the entry site corresponded with ulcer-like projections that had been pointed out at the thrombosed false lumen. One patient presented with moderate back pain when the false lumen changed, while the others were symptom free. During the follow-up period, 2 patients died of rupture of the false lumen. One patient was treated with a surgical operation. The patent false lumen became re-thrombosed in 1 patient. Although the majority of cases of thrombosed aortic dissection are considered to have a good prognosis, some cases change to the patent type without clinical symptoms. Radiologists should be aware of the unstable nature of the thrombosed type aortic dissection, and follow-up examinations should be performed at least once a week for three weeks after the onset of dissection.

Acute Disease↗

[Three-dimensional imaging of portal and hepatic veins using helical CT with intravenous injection of contrast medium].

The authors developed a simple new method for obtaining three-dimensional helical CT images that simultaneously depict both the portal and hepatic veins. One hundred ml of contrast medium was injected intravenously, and helical CT was carried out. Three-dimensional venous images were successfully displayed with rotational movement on the CRT monitor. The new method provided high-quality images in which the anatomical relationship between the portal and hepatic venous systems could be clearly visualized. This information was useful in helping angiographers to successfully perform transjugular intrahepatic portosystemic shunt and will be of great value for the presurgical planning of hepatic resection.

Adult↗

The role of insulin in activation of two enhancers in the mouse GLUT1 gene.

We identified two enhancer elements of the mouse GLUT1 gene responsive to serum, growth factor, and oncogenes; the first enhancer element (enhancer-1) is located 2.7 kilobases upstream of the cap site of the gene, and the second one (enhancer-2) is located in the second intron of the gene (Murakami, T., Nishiyama, T., Shirotani, T., Shinohara, Y., Kan, M., Ishii, K., Kanai, F., Nakazuru, S., and Ebina, Y. (1992) J. Biol. Chem. 267, 9300-9306). In the present work, we describe the role of insulin in activation of these two enhancers. NIH/3T3 HIR3.5 cells, which express a large number of insulin receptors, were stably transformed by hybrid genes containing the enhancer(s) and promoter of GLUT1 gene and the coding region of chloramphenicol acetyltransferase (CAT) gene as a reporter gene. In stable transformants of the reporter gene without the enhancers, the CAT mRNA was not induced by insulin; however, in clones containing the reporter gene with enhancer-1, the CAT mRNA was induced by insulin at 30 min and reached a maximum at 1 h. In clones transfected by the reporter gene with enhancer-2, the CAT mRNA was induced at 1 h and reached a maximum at 3 h. To determine the early response element to insulin in enhancer-1, transformants of hybrid reporter genes containing truncated or mutated enhancer-1 were examined. The homologous sequence with the serum response element in enhancer-1 is essential for an early response to insulin.

3T3 Cells↗

NMR studies of 1H resonances in the 10-18-ppm range for aspartate aminotransferase from Escherichia coli.

We have recorded 500-MHz 1H NMR spectra in the 10-18-ppm range for aspartate aminotransferase from Escherichia coli and for three specific mutant forms. Histidine 143 has been replaced by either alanine or asparagine. In the third mutant, tryptophan 140 has been replaced by phenylalanine. The NMR spectrum of the native enzyme is very similar to that of porcine cytosolic aspartate aminotransferase in the most downfield region. However, the resonances of the proton on the ring nitrogen of the pyridoxal 5'-phosphate (peak A) and on the His-143 imidazole ring (peak B) of the E. coli enzyme are broader and more readily lost at low pH or higher temperatures than those of the porcine enzyme. The possible role of tautomerism in promoting such broadening is discussed. In the histidine mutant proteins, peak A of the pyridoxal 5'-phosphate form is too broad to see under most conditions but is clearly present in the pyridoxamine phosphate form. Peak B is missing in the 2 histidine mutants. Observation of nuclear Overhauser effects further confirms the identity of B as the resonance of HN epsilon 2 of His-143 and that of peak D at approximately 11.8 ppm as HN epsilon 2 of His-189. The mutant spectra also provide insight into electronic interactions between groups in and near the active site which confirm and supplement conclusions drawn from spectra of porcine cAspAT. While no clear loss of a peak was observed for the Trp-140 mutant in its free form, the spectrum of the succinate complex lacked a strong band at 11.26 ppm. This may represent the Trp-140 indole NH proton which has been shifted downfield by binding to a succinate carboxylate group. While our results confirm the basic similarity of cytosolic aspartate aminotransferase and E. coli aspartate aminotransferase 1H NMR spectra, they also point out differences that may be useful in identifying resonances. A large number of mutant proteins have been prepared for the E. coli enzyme. The present results provide essential information for future study of these mutants and for study of NMR spectra of isotopically labeled enzyme.

Alanine↗

Simultaneous measurement of nicotinic acid and its major metabolite, nicotinuric acid in urine using high-performance liquid chromatography: application of solid-liquid extraction.

The concentrations of nicotinic acid (NiAc) and nicotinuric acid (NiUAc), a major metabolite of NiAc, were simultaneously determined in urine using solid-phase extraction (cation-exchange extraction) and reversed-phase high-performance liquid chromatography with ultraviolet detection. The intra- and inter-day precision studies showed good reproducibilities: the coefficients of variations were less than 8.1% for NiAc and 8.8% for NiUAc. The calibration curves were linear (r2 > 0.9934) in the concentration range 10-1000 micrograms/ml. The removal of endogenous interferences in urine by solid-phase extraction presented here is superior to the pretreatment protocols reported previously by other workers. The method was used in a preliminary pharmacokinetic study in rats after intravenous administration of NiAc (5 and 15 mg/kg).

Animals↗

Auxin inducibility and developmental expression of axi 1: a gene directing auxin independent growth in tobacco protoplasts.

We describe the characterization of axi 1, a tobacco gene isolated by activation T-DNA tagging which apparently plays a role in auxin action. Upon deregulated expression, axi 1 confers on protoplasts the ability to grow in culture not only in the absence of auxin but also in high auxin concentrations where maximal frequencies of cell division are not observed in wild-type protoplasts. In wild-type plants axi 1 is transcribed principally in root tissue. In the tagged plant line, axi 159, axi 1 RNA can be detected in all tissues tested. Freshly isolated wild-type protoplasts require auxin for the accumulation of detectable levels of axi 1 transcript and this precedes maximal levels of cell division. In contrast, axi 1 RNA appears in protoplasts isolated from axi 159 plants in the absence of auxin. axi 1 was localized to 6.2 kb of plant genomic DNA flanking the right T-DNA border sequence. axi 1 is interrupted by nine introns and in tobacco it is a member of a small gene family. Database searching reveals no similarity within the coding region with other genes. Sequences within the fourth intron are similar to those located in the non-coding regions of other plant genes, some of which are known to be auxin inducible. A DNA fragment containing the conserved sequence acts as an auxin responsive element in transient expression assays in wild-type protoplasts and this response is higher in axi 159 protoplasts. This suggests that auxin induced axi 1 expression may be mediated by a region contained within an intron sequence and that the axi 1 product might play a role in this induction.

Base Sequence↗

Oxygen free radicals and calcium homeostasis in the heart.

Many experiments have been done to clarify the effects of oxygen free radicals on Ca2+ homeostasis in the hearts. A burst of oxygen free radicals occurs immediately after reperfusion, but we have to be reminded that the exact levels of oxygen free radicals in the hearts are yet unknown in both physiological and pathophysiological conditions. Therefore, we should give careful consideration to this point when we perform the experiments and analyze the results. It is, however, evident that Ca2+ overload occurs when the hearts are exposed to an excess amount of oxygen free radicals. Through ATP-independent Ca2+ binding is increased, Ca2+ influx through Ca2+ channel does not increase in the presence of oxygen free radicals. Another possible pathway through which Ca2+ can enter the myocytes is Na(+)-Ca2+ exchanger. Although, the activities of Na(+)-K+ ATPase and Na(+)-H(+) exchange are inhibited by oxygen free radicals, it is not known whether intracellular Na(+) level increases under oxidative stress or not. The question has to be solved for the understanding of the importance of Na(+)-Ca2+ exchange in Ca2+ influx process from extracellular space. Another question is 'which way does Na(+)-Ca2+ exchange work under oxidative stress? Net influx or efflux of Ca2+?' Membrane permeability for Ca2+ may be maintained in a relatively early phase of free radical injury. Since sarcolemmal Ca(2+)-pump ATPase activity is depressed by oxygen free radicals, Ca2+ extrusion from cytosol to extracellular space is considered to be reduced. It has also been shown that oxygen free radicals promote Ca2+ release from sarcoplasmic reticulum and inhibit Ca2+ sequestration to sarcoplasmic reticulum. Thus, these changes in Ca2+ handling systems could cause the Ca2+ overload due to oxygen free radicals.

Animals↗

Purification and characterization of a human erythrocyte-derived growth factor with a wide target cell spectrum.

A cell extract from human erythrocytes promoted the growth of a wide variety of cell types, namely human and mouse myeloid cells, human and mouse T cells, human B cells, human melanoma cells, mouse transformed fibroblast cells, mouse mastocytoma cells, human lung fibroblast cells, and mouse bone marrow fibroblast/stroma-like cells. The growth-promoting activity was acid- and heat-labile and sensitive to proteases, indicating the proteinaceous nature of the molecule. The activity was also lost upon exposure to 2-mercaptoethanol. The novel growth-promoting factor, termed basic growth factor because of its fundamental effect and a wide target cell spectrum, was purified by sequential anion-exchange, hydrophobic, gel filtration, hydroxylapatite, and reverse-phase high performance liquid chromatographies. The purified factor has an apparent molecular mass of 53 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing and nonreducing conditions. The factor migrated at 270 kDa on native gradient polyacrylamide gel electrophoresis. Therefore, the factor consists of a homopolymer of a single polypeptide chain. The purified factor promoted the growth of the same cell types as the cell extract, except for bone marrow cells.

Amino Acids↗

[Evaluation of bone marrow by opposed phase T1-weighted images and enhanced MR imaging].

We investigated bone marrow in a control group, cases of aplastic anemia and post-irradiation patients by examining T1-weighted (T1WI), short TI inversion recovery (STIR), opposed phase T1WI (op-T1WI) and Gd-DTPA enhanced op-T1WI images obtained by 0.5T MRI. Bone marrow was classified into four types based on MR findings. Normal marrow showed low intensity on op-T1WI and STIR images without enhancement (I). Fatty marrow, which showed high intensity on T1WI and op-T1WI images was observed in aplastic anemia and post-irradiation patients (II). Hematopoietic marrow (III) showed low intensity on op-T1WI and enhanced, while active hematopoietic marrow (IV) revealed high intensity on both STIR and op-T1WI images and was enhanced following Gd-DTPA infusion. Aplastic anemia of moderate grade included types II, III and IV. Enhanced MR was needed to differentiate between types I and III since both types showed low intensity on op-T1WI images. Furthermore, type IV was considered as hyperplastic compared with type III. Enhanced MR and op-T1WI images were useful in evaluating hematopoiesis of bone marrow.

Adolescent↗

Simultaneous measurement of [Na+]i and Ca2+ transients in an isolated myocyte: effects of strophanthidin.

We have developed a new method to measure [Na+]i and Ca2+ transients of a beating cardiac myocyte using a Na(+)-sensitive fluorescent probe, sodium-binding benzofuran isophthalate (SBFI) and a Ca(2+)-sensitive fluorescent probe, fluo-3. There was no interaction between two probes, and the artifact due to contraction could be eliminated in the measurement of [Na+]i. [Na+]i in guinea pig ventricular myocytes stimulated at 1 Hz was 8.0 +/- 0.7mM. Strophanthidin (10 microM), initially, increased the amplitude and the basal level of Ca2+ transients in association with an increase in [Na+]i. When arrhythmias were induced, the amplitude of Ca2+ transients decreased while [Na+]i and the basal level of Ca2 transients continued to increase. These results suggested that the diastolic [Ca2+]i was closely related to [Na+]i. However, the systolic [Ca2+]i, which could be influenced by other factors, was dissociated from [Na+]i in the condition of Ca2+ overload.

Aniline Compounds↗

Site-directed mutagenesis of the histamine H1 receptor: roles of aspartic acid107, asparagine198 and threonine194.

Based on structural comparison with other biogenic amine receptors and the histamine H2 receptor, it has been suggested that in the human histamine H1 receptor, Asp107, Thr194, and Asn198 are the residues involved in binding of histamine. We therefore used site-directed mutagenesis to investigate the roles of these three amino acid residues. Asp107 was essential for both agonist and antagonist binding. Asn198 was necessary for agonist but not for antagonist binding. Thr194 was not important for either type of binding. A good correlation was found between agonist binding and receptor activation for all the wild-type and mutant receptors. The results show that the histamine H1 receptor recognizes and is activated by histamine through the interactions of Asp107 and the amino group, and Asn198 and the imidazole ring.

Amino Acid Sequence↗

Ultrasonographic diagnosis of pyelonephritis in a cow.

A 2-year-old Holstein cow was examined because of a 2-month history of anorexia, decreased milk production, and a white vaginal discharge. Results of physical, laboratory, and ultrasound examinations were consistent with pyelonephritis. Through ultrasound examination, the right kidney and ureter were identified as structurally involved. Right nephrectomy was performed. Eight months after nephrectomy, the cow was clinically normal. In this cow, ultrasonography helped to determine disease severity, to direct the course of treatment, and to improve the accuracy of diagnosis and prognosis.

Animals↗