Search PubMed⌕ Search

Biomedical subjects

H Hayashi

Publications and source records attributed to H Hayashi.

At least 577 records · Page 32Linked to original sources

Recording of electroencephalograms and electrocardiograms during daytime sleep in trained canines: preparation of the sleeping dogs.

Although respiration in trained canines is well investigated, the process of preparing dogs has not been described in any great detail. Moreover, their daytime patterns of sleep and wakefulness during 1 or 2 h of electroencephalogram (EEG) and electrocardiogram (ECG) recordings are not clear. Therefore, we describe the process of selecting and training dogs, in which we recorded EEG and ECG in the laboratory. First, 14 of 1242 dogs dealt with over a 1 year period were chosen. They were trained for 2 h to lie quietly and to sleep in the laboratory; this training procedure was repeated 152 times. Three dogs were then selected and a permanent tracheostomy was performed in one. Finally, EEG and ECG were recorded with the bipolar fine needle electrodes; respiration was recorded simultaneously through a tube inserted to a tracheostomy in one dog. Wakefulness, slow wave sleep (SWS) and rapid eye movement (REM) sleep (REMS) were identified according to the EEG pattern and on the basis of the behavioral criteria. Recordings were performed 12 or 13 times in each dog. Complete sleep cycles, including wakefulness, SWS and REMS in this sequence, were observed 3.9-4.1 times. The mean duration of SWS was 2.2-4.4 min and that of REMS was 3.5-4.6 min. The REMS latency was 33.9-41.8 min. Fluctuation of heart rate with respiration, termed respiratory sinus arrhythmia, was noted in the ECG. Heart beat increased with inspiration and decreased with expiration. The present study demonstrates how to select and train sleeping dogs and shows their undisturbed daytime sleep and wakefulness patterns.

Animals↗

Adsorption of hepatitis C virus particles onto the dialyzer membrane.

It was recently found that the blood level of hepatitis C virus (HCV) RNA is significantly reduced after each dialysis procedure in patients on chronic hemodialysis. This study was designed to elucidate the mechanism for this phenomenon. In two patients with high serum levels of HCV RNA, the filtrate through the dialyzer (TF-alpha, Teijin Co., Tokyo, Japan) was analyzed for viral RNA using the polymerase chain reaction. At the end of dialysis, the filter was washed with saline, and during the saline washing, aliquots were taken for quantification of RNA by the branched DNA method. The HCV core antigen was quantified as a measure of viral particles, and hemoglobin was also measured for correcting for blood contamination. After the clearance of the blood, the filter was washed with guanidinium isothiocyanate, and the recovery of RNA was measured. The filtrate did not contain detectable RNA. The saline washing of the filter after dialysis contained a significant amount of RNA. Washing with guanidinium isothiocyanate of the thoroughly saline washed filter also recovered a significant amount of RNA. During saline washing, the recovery of RNA in the washing was much delayed behind that of hemoglobin, suggesting the adsorption of the former onto the filter membrane. There was a discordant recovery of RNA and HCV core antigen in the washing, the recovery of the former being delayed behind that of the latter. These results indicate that viral particles are adsorbed onto the inner surface of the filter membrane during dialysis. Some of these adsorbed viral particles are perhaps destroyed by hydraulic pressure applied to blood for dialysis.

Adsorption↗

Effects of inhaled nitric oxide on postoperative pulmonary circulation in patients with congenital heart disease.

We studied 22 patients with residual pulmonary hypertension or symptoms of postoperative pulmonary hypertensive crisis. They received low-dose inhalation (10 ppm) of nitric oxide (NO), a selective pulmonary vasodilator, after total correction for congenital heart anomalies. Fifteen minutes of NO inhalation improved the pulmonary circulation and lessened the imbalance in the ventilation-perfusion ratio in both groups. Thus, NO inhalation is effective in the treatment of pulmonary hypertension and in the prevention of pulmonary hypertensive crises after total correction for congenital heart anomalies. All patients continued to receive NO therapeutically. The duration of such therapeutic NO inhalation was well correlated with postoperative Qp/Qs (p = 0.014) and Rp/Rs (p = 0.029).

Administration, Inhalation↗

Long-term sedation with isoflurane in postoperative intensive care in cardiac surgery.

After cardiac surgery, patients often require prolonged mechanical ventilation. We studied the effectiveness and potential toxicity of isoflurane sedation in 40 patients undergoing mechanical ventilation after cardiovascular surgery. All patients who received isoflurane (0.5-1.0 minimum alveolar concentration [MAC] were well sedated by it without significant adverse effects, such as renal, hepatic, or cardiovascular dysfunction. The highest serum inorganic fluoride concentration recorded was 45 mumol/L after 98 MAC h. Patients on isoflurane recovered more rapidly and were weaned from mechanical ventilation sooner than those sedated with intravenous drugs including fentanyl/midazolam. Patients who received intravenous sedatives, but not those on isoflurane, often showed tachyphylaxis in the early stages, and some exhibited an abstinence syndrome involving nonpurposeful movements. Patients sedated with isoflurane did not show these two side effects. In conclusion, isoflurane can provide effective long-term sedation for patients after cardiovascular surgery without significant adverse effects.

Acute Kidney Injury↗

Helium/oxygen breathing improves hypoxemia after cardiac surgery.

The effects of helium/oxygen (He/O2) on oxygenation (respiratory index [PaO2/FiO2] and intrapulmonary shunt [Qs/Qt]) and on lung parameters (dynamic compliance [Cdyn] and peak inspiratory pressure [PIP]) were studied in 12 patients. After cardiac surgery, they had impairment of oxygenation without physiological findings and with normal chest radiographs despite having a positive end expiratory pressure of up to 10 cm H2O. After 90 min of inhalation of He/O2, oxygenation had improved significantly; that is, PaO2/FidO2 increased significantly (from 113 to 174 mm Hg; mean values are given; p < 0.01), and there was a significant fall in Qs/Qt (from 29 to 19%; p < 0.001) together with an increase in Cdyn (from 60 ml/cm H2O to 64 ml/cm H2O; p < 0.05). These results suggest that He/O2 may have improved oxygenation by recruiting previously obstructed small airways and alveoli.

Administration, Inhalation↗

Successful treatment of mediastinitis after cardiovascular surgery using electrolyzed strong acid aqueous solution.

Dilute povidone-iodine solution has been widely used as an irrigant for the treatment of mediastinitis. However, its use is not without adverse effects and often causes poor growth of granulation tissues. To avoid the problems seen with the use of povidone-iodine solution, we applied electrolyzed strong acid aqueous solution (ESAAS) to mediastinal irrigation in 4 patients (2 infants and 2 adults) who developed mediastinitis after cardiovascular surgery. According to the "open" method, the mediastinal wound was left open and irrigated with ESAAS 1 to 3 times a day until the infection was eradicated. Satisfactory growth of granulation tissues was observed in all patients treated with no evidence of adverse effects attributable to ESAAS. Delayed primary sternum closure was performed for 2 patients, and musculocutaneous transposition of rectus abdominis for 1. Our experience suggests that irrigation with ESAAS is a safe and effective method of therapy for mediastinitis.

Acids↗

Role of sympathovagal balance in the initiation of idiopathic ventricular tachycardia originating from right ventricular outflow tract.

VT originating from the right ventricular outflow tract (RVOT) is prone to occur when sympathetic nervous activity is increased. beta-Blockade is, therefore, effective in suppressing this VT. The purpose of this study was to determine the role of sympathovagal balance assessed by heart rate variability (HRV) in the spontaneous initiation of repetitive premature ventricular contractions (PVCs) and VT (five or more consecutive PVCs) arising from RVOT in seven patients without structural heart diseases. Frequency-domain measures of HRV were determined by analyzing 24-hour Holter electrocardiographic recording with the maximum entropy method over a 1,280-second period immediately before the onset of 35 single PVCs, 26 episodes of 2-4 consecutive PVCs, and 21 episodes of VT. High frequency component (HF: 0.15-0.40 Hz) was used as an index of parasympathetic activity, and the ratio of low frequency component (LF: 0.04-0.15 Hz) to HF (LF/HF ratio), as an index of sympathovagal balance. NN50(%), a time-domain variable of parasympathetic activity, was also determined. Mean RR interval and any measures of HRV did not change significantly before single PVCs. Mean RR interval shortened and HF decreased prior to repetitive PVCs and VT. The LF/HF ratio, however, increased only before the onset of VT. NN50(%) tended to decrease before repetitive PVCs and decreased significantly before VT. With propranolol (30-60 mg/day), frequency of repetitive PVCs was suppressed from 2,048 +/- 1,201 to 746 +/- 658/day and VT was totally abolished, but frequency of single PVCs did not change significantly. In conclusion, sympathetic predominance plays an important role in the initiation of repetitive PVCs and VT originating from RVOT in patients without structural heart diseases.

Adolescent↗

Removal of human immunodeficiency virus type 1 (HIV-1) protease inhibitors from preparations of immature HIV-1 virions does not result in an increase in infectivity or the appearance of mature morphology.

The processing of gag and gag-pol polyproteins by human immunodeficiency virus type 1 (HIV-1) protease is a crucial step in the formation of infectious HIV-1 virions. In this study, we examine whether particles produced in the presence of inhibitors of HIV-1 protease can subsequently undergo gag polyprotein cleavage with restoration of infectivity following removal of the inhibitors. Viral particles produced during 7 days of culture in the presence of the protease inhibitors KNI-272 (10 microM) and saquinavir (5 microM) contained predominantly p55gag polyprotein but little or no p24gag cleavage product. Following resuspension of the particles in medium free of the inhibitor, some gag polyprotein processing was detected in particles produced from the KNI-272-treated cells, but not from the saquinavir-treated cells within the first 3 h. However, the majority of the protein remained as p55gag throughout a 48-h experimental period. The infectivity (50% tissue culture infective dose per milliliter) of the viral particles from KNI-272-treated cells was 10(6)-fold lower than that of control particles and did not significantly increase over the 48 h after the inhibitor was removed, despite the apparent return of protease function in a subset of these virions. This failure to restore infectivity was due neither to a reduction in the number of particles produced by protease inhibitor-treated cells nor to a failure of HIV RNA to be packaged in the virions. These particles also failed to express the mature phenotype by electron microscopy. Thus, while some processing of the gag polyprotein can occur in isolated HIV virions, this does not appear to be sufficient to restore infectivity in the majority of particles. This finding suggests that there may be constraints on postbudding polyprotein processing in the production of viable particles. These results should have positive implications regarding the use of protease inhibitors as anti-HIV drugs.

Blotting, Western↗

Cloning of genes coding for L-sorbose and L-sorbosone dehydrogenases from Gluconobacter oxydans and microbial production of 2-keto-L-gulonate, a precursor of L-ascorbic acid, in a recombinant G. oxydans strain.

We have purified L-sorbose dehydrogenase (SDH) and L-sorbosone dehydrogenase (SNDH) from Gluconobacter oxydans T-100 that showed an ability to convert D-sorbitol to 2-keto-L-gulonate (2-KLGA). A genomic library of Gluconobacter oxydans T-100 was screened with a probe, a 180-bp PCR product which was obtained from degenerate oligodeoxyribonucleotides based on the elucidated sequence of the purified SDH (used as primers) and the genomic DNA of G. oxydans T-100 (used as a template). From sequencing of the DNA from a clone positive to the probe, the SNDH and the SDH were estimated to be coded in sequential open reading frames with 1,497 and 1,599 nucleotides, respectively, which was confirmed by expression of the DNA in Escherichia coli that showed both enzymatic activities. The DNA was introduced to a shuttle vector which was prepared from a plasmid of G. oxydans T-100 and pHSG298 to obtain an expression vector designated pSDH155. The production of 2-KLGA by pSDH155 in G. oxydans G624, an L-sorbose-accumulating strain, was improved to 230% compared to that of G. oxydans T-100. Chemical mutation of the host strain to suppress the L-idonate pathway and replacement of the original promoter with that of E. coli tufB resulted in improving the production of 2-KLGA. Consequently, high-level production from D-sorbitol to 2-KLGA (130 mg/ml) was achieved by simple fermentation of the recombinant Gluconobacter.

Acetobacteraceae↗

Exchange of glutamate and gamma-aminobutyrate in a Lactobacillus strain.

Lactobacillus sp. strain E1 catalyzed the decarboxylation of glutamate (Glu), resulting in a nearly stoichiometric release of the products gamma-aminobutyrate (GABA) and CO2. This decarboxylation was associated with the net synthesis of ATP. ATP synthesis was inhibited almost completely by nigericin and about 70% by N,N'-dicyclohexylcarbodiimide (DCCD), without inhibition of the decarboxylation. These findings are consistent with the possibility that a proton motive force arises from the cytoplasmic proton consumption that accompanies glutamate decarboxylation and the electrogenic Glu/GABA antiporter and the possibility that this proton motive force is coupled with ATP synthesis by DCCD-sensitive ATPase.

Adenosine Triphosphate↗

Sequence of xynC and properties of XynC, a major component of the Clostridium thermocellum cellulosome.

The nucleotide sequence of the Clostridium thermocellum F1 xynC gene, which encodes the xylanase XynC, consists of 1,857 bp and encodes a protein of 619 amino acids with a molecular weight of 69,517. XynC contains a typical N-terminal signal peptide of 32 amino acid residues, followed by a 165-amino-acid sequence which is homologous to the thermostabilizing domain. Downstream of this domain was a family 10 catalytic domain of glycosyl hydrolase. The C terminus separated from the catalytic domain by a short linker sequence contains a dockerin domain responsible for cellulosome assembly. The N-terminal amino acid sequence of XynC-II, the enzyme purified from a recombinant Escherichia coli strain, was in agreement with that deduced from the nucleotide sequence although XynC-II suffered from proteolytic truncation by a host protease(s) at the C-terminal region. Immunological and N-terminal amino acid sequence analyses disclosed that the full-length XynC is one of the major components of the C. thermocellum cellulosome. XynC-II was highly active toward xylan and slightly active toward p-nitrophenyl-beta-D-xylopyranoside, p-nitrophenyl-beta-D-cellobioside, p-nitrophenyl-beta-D-glucopyranoside, and carboxymethyl cellulose. The Km and Vmax values for xylan were 3.9 mg/ml and 611 micromol/min/mg of protein, respectively. This enzyme was optimally active at 80 degrees C and was stable up to 70 degrees C at neutral pHs and over the pH range of 4 to 11 at 25 degrees C.

Amino Acid Sequence↗

In situ localization of basic fibroblast growth factor protein and mRNA in the retina.

Basic fibroblast growth factor (FGF) protein has been recognized as a potent factor for angiogenesis and as a mitogen. The sites of basic FGF in the mammalian retina have varied from report to report. On the other hand, only the inner segments of the photoreceptor cells have been reported to synthesized basic FGF as revealed by the presence of mRNA for basic FGF by in situ hybridization. To define the sites of basic FGF and its mRNA in the human retina, we immunohistochemically localized basic FGF protein and mRNA for basic FGF by in situ hybridization in normal human retinas. Basic FGF protein was found in the ganglion cell layer, the inner and outer nuclear layers, and the basement membranes of Müller cells (the inner limiting membrane), blood vessels, and the retinal pigment epithelium (Bruch's membrane). The mRNA for basic FGF was found in the cells of the ganglion cell layer, the inner nuclear layer, and the outer nuclear layer, and the inner segments of the photoreceptor cells. These findings suggest that basic FGF in the human retina functions in both an autocrine as well as a paracrine fashion.

Fibroblast Growth Factor 2↗

Nocturnal blood pressure fall on ambulatory monitoring in a large international database. The "Ad Hoc' Working Group.

A wide range of definitions is used to distinguish subjects in whom blood pressure (BP) falls at night (dippers) from their counterparts (nondippers). In an attempt to standardize the definition of nondipping, we determined the nocturnal BP fall and night-day BP ratio by 24-hour ambulatory monitoring in 4765 normotensive and 2555 hypertensive subjects from 10 to 99 years old. In all subjects combined, the systolic/diastolic nocturnal fall and corresponding ratio averaged (+/- SD) -16.7 +/- 11.0/ -13.6 +/- 8.1 mm Hg and 87.2 +/- 8.0%/83.1 +/- 9.6%, respectively. In normotensive subjects, the 95th percentiles were -0.3/-1.1 mm Hg for the nocturnal fall and 99.7%/98.3% for the night-day ratio. Both the fall and ratio showed a curvilinear correlation with age. The smallest fall and largest ratio were observed in older (> or = 70 years) subjects. A higher BP on conventional sphygmomanometry was associated with a larger systolic (partial r = .11) and diastolic (r = .12) nocturnal BP fall. The diastolic (r = .08) but not the systolic night-day ratio increased with higher conventional BP. The nocturnal BP fall was larger and the corresponding night-day ratio smaller in oscillometric (n = 5884) than in auscultatory (n = 1436) recordings, in males (n = 3730) than in females (n = 3590), and in Europe (n = 4556) than in the other continents (n = 2764). The distributions of the nocturnal BP fall and night-day ratio showed considerable overlap among normotensive and hypertensive subjects, but the overlap tended to be larger for the ratio than for the fall. Of all subjects, 3.2% had systolic and diastolic ratios of 100% or more. With adjustments applied for confounders, the probability of being a nondipper increased 2.8 times (95% confidence interval, 2.0-4.0) from 30 to 60 years and 5.7 times (4.4-7.4) from 60 to 80 years. The odds ratios were 1.0 (0.8-1.4) for males versus females. 1.6 (1.2-2.1) for subjects with definite hypertension versus normotensive subjects, 2.4 (1.2-4.7) for Asians (n = 2213, 96% Japanese) versus inhabitants of the other continents, and 2.4 (1.5-3.8) for subjects examined with auscultatory versus oscillometric devices. In conclusion, the mathematical definition of nondipping, ie, having a night-day ratio of 100% or more for systolic and diastolic BPs, closely approximated the 95th percentiles of the night-day ratio in normotensive subjects. The ratio depends less on BP level than the nocturnal BP fall and is therefore to be preferred in the definition of dipping status. Notwithstanding the present findings, the reproducibility of nondipping and its prognostic significance need further clarification.

Adolescent↗

Production and characterization of specific anti-peptide antiserum against free alpha-subunit of rat pituitary glycoprotein hormones.

To obtain an antibody specific for the alpha-subunit of rat pituitary glycoprotein hormones, we synthesized a peptide corresponding to the sequence 37-53 (ST-7: Phe-Ser-Arg-Ala-Tyr-Pro-Thr-Pro-Ala-Arg-Ser-Lys-Lys-Thr-Met-Leu-Val) of the rat alpha-subunit. The polyclonal antiserum against this peptide was generated in rabbits. This region is hydrophilic and highly conserved among several mammalian species. Noncompetitive binding tests showed that the ST-7 antiserum had specific affinity for the rat free alpha-subunit, but not for rat intact LH, FSH, and TSH. The ST-7 antiserum immunostained two types of cells in the rat anterior pituitary, i.e., gonadotrophs and thyrotrophs. This was also the case in mouse, cattle, sheep, and pig, which have an identical sequence of ST-7 in their alpha-subunit. The pituitary cells of horse (Arg substituted for Lys as residue 48 of the rat alpha-subunit), human, and eel (Leu for Ala at residue 45), chicken (Met for Ala at residue 45), and bullfrog (Tyr for Phe at residue 37 and Met for Ala at residue 45) were not stained with the ST-7 antiserum. This study indicated that the ST-7 antiserum is sequence-specific for the alpha-subunit and is therefore useful for immunohistochemical studies on the secretory pathway of the free alpha-subunit.

Amino Acid Sequence↗

DNA single-strand breaks in L-132 cells resulting from inhibition of repair polymerization shortly after exposure to dimethylarsinic acid.

DNA single-strand breaks due to the inhibition of repair polymerization in cultured human pulmonary epithelial (L-132) cells after exposure to dimethylarsinic acid (DMAA), a main metabolite of inorganic arsenics in mammals, were examined. The strand breaks were detected by an alkaline elution method with the use of inhibitors of DNA polymerase, aphidicolin (aph) and 2',3'-dideoxythymidine (ddT); the former inhibits DNA polymerases alpha, delta and epsilon, and the latter inhibits DNA polymerase beta. Generally, DNA polymerases delta and epsilon are thought to be associated with necleotide excision (long patch) repair and polymerase beta with base excision (short patch) repair. After exposure of the L-132 cells to 10 mM DMAA, the breaks occurred in a time-dependent manner during incubation for 1-6 h under the inhibition of aph-sensitive polymerases with 50 micrograms/ml aph plus 10 mM hydroxyurea (HU) for the last 1 h of the DMAA exposure. Also, when DNA polymerase beta was inhibited with 10 mM ddT plus 1 microM methotrexate (MTX), the exposure of L-132 cells to 10 mM DMAA for 6 h significantly induced DNA single-strand breaks. An experiment of the co-treatment with both aph and ddT suggested that in the DNA repair process, aph-sensitive polymerases, probably polymerases delta and/or epsilon, and polymerase beta, functioned independently on different lesions induced after exposure to DMAA.

Animals↗

Effect of gemfibrozil on centrifugal behavior of rat peroxisomes and activities of peroxisomal enzymes involved in lipid metabolism.

The effect of gemfibrozil, and analogue of clofibric acid, on the centrifugal behavior of peroxisomes and activity of peroxisomal enzymes involved in lipid metabolism was studied. Rats were fed chow containing 0.2% gemfibrozil for 2 weeks. Nycodenz gradient centrifugation of the light mitochondrial fraction revealed that peroxisomes of gemfibrozil-treated rats were concentrated in fractions of higher density compared with control rats. The activity of fatty acyl-CoA oxidase, crotonase, beta-hydroxybutyryl-CoA dehydrogenase, and thiolase (individual enzymes of the peroxisomal fatty acid beta-oxidation system) were enhanced 9.6, 2.3, 3.4 and 9.1 times respectively compared with controls, by treatment. The hydroxymethylglutaryl-CoA (HMG-CoA) reductase (rate-limiting enzyme of cholesterol synthesis) activity of peroxisomes and microsomes was greatly increased by in vivo treatment with gemfibrozil, but was decreased by addition of the agent to the assay mixture of the enzyme. Gemfibrozil directly inhibited the reductase activity and did so at a lower concentration than clofibric acid. Peroxisomal reductase was more resistant to damage by the agent than the microsomal enzyme. The HMG-CoA reductase activity of peroxisomes and microsomes of hyperlipidemic rats was also increased by in vivo treatment with gemfibrozil, whereas the serum cholesterol level was hardly changed. These results indicate that the effect of gemfibrozil differs from that of clofibric acid, the main difference being the effect on HMG-CoA reductase. Gemfibrozil increased reductase activity in vivo, unlike clofibric acid, but inhibited the enzyme in vitro to a greater extent than clofibric acid.

Animals↗

Effects of nebracetam on synaptosomal monoamine uptake of striatal and hippocampal regions in rats.

Effects of nebracetam, a novel nootorpic agent, on the synaptosomal uptake of neurotransmitter monoamines of the brain regions were examined. Striatal and hippocampal synaptosomes were isolated by the Percoll gradient method, and the striatal dopamine uptake and hippocampal serotonin uptake were measured in the presence of different concentrations (1 to 1000 microM) of nebracetam in vitro. A significant reduction in dopamine uptake in the striatum and serotonin uptake in the hippocampus was seen at concentrations of 100 microM or above. In in vivo microdialysis study, there were no appreciable changes in the extracellular concentrations of striatal dopamine and hippocampal serotonin when this agent at a dose of 30 mg/kg, which was effective in improving ischemic brain energy metabolism, was applied i.p. to the rat. The ineffectiveness of nebracetam in the in vivo microdialysis may be due to low levels of the concentration of nebracetam when the agent was administered i.p. at a dose of 30 mg/kg, since the brain blood concentration of this agent is pharmacokinetically estimated to be no more than 15 microM when this dose of nebracetam is employed. Thus, it is unlikely that this agent at a pharmacologically effective dose alters dopamine or serotonin uptake in the brain nerve terminal under normal conditions.

Animals↗