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Biomedical subjects

Y Miyamoto

Publications and source records attributed to Y Miyamoto.

At least 19 recordsLinked to original sources

Determination of urinary beta-phenylethylamine as its N-benzenesulphonamide derivative by gas chromatography with flame photometric detection.

A selective and sensitive method for the determination of urinary beta-phenylethylamine (PEA) by gas chromatography (GC) has been developed. After extraction of the urine sample with n-pentane, PEA was converted into its N-benzenesulphonamide derivative and then determined by GC with flame photometric detection using a DB-1301 capillary column. By using this method, nanogram amounts of PEA in urine could be accurately and precisely determined without any influence from coexisting substances. Analytical results for the determination of PEA in urine samples from normal subjects are presented.

Chromatography, Gas

The gamma-aminobutyric acid transporter and its interaction with taurine in the apical membrane of the bovine retinal pigment epithelium.

The characteristics of gamma-aminobutyric acid (GABA) uptake were investigated in apical membrane vesicles prepared from the bovine retinal pigment epithelium. An inwardly directed NaCl gradient stimulated GABA uptake markedly, and the time course of uptake exhibited an overshoot phenomenon indicating the presence of an active transport mechanism for GABA in these membranes. Other monovalent cations were not capable of substituting for Na+. In addition to this obligatory requirement for Na+, the GABA uptake also exhibited a Cl(-)-dependence, evident from the observations that the uptake was negligible in the presence of NaF or sodium gluconate in place of NaCl. NO3- and SCN- could substitute for Cl- to some extent. The uptake process was electrogenic, with a Na+/Cl-/GABA stoichiometry of 2:1:1 or 3:1:1. Substrate-specificity studies showed that the beta-amino acids such as taurine, hypotaurine and beta-alanine interacted with the GABA uptake process. Uptake of GABA could be completely inhibited by an excess of taurine and, similarly, uptake of taurine could be completely inhibited by an excess of GABA, suggesting that common transport processes operate in the uptake of these two compounds. However, a number of compounds which are specific inhibitors of GABA uptake inhibited taurine uptake only to a maximum of 50%. Kinetic analysis of GABA uptake in the concentration range 0.1-10 microM revealed that the uptake occurred via a single system and that taurine was a competitive inhibitor of this system. The Michaelis-Menten constant (Kt) for GABA was 0.94 microM and the apparent inhibition constant (Ki) for taurine was 230 microM. On the contrary, even though the kinetic analysis of taurine uptake in the concentration range 25-150 microM revealed participation of a single system in the uptake process, the inhibition of taurine uptake by GABA was not competitive. The presence of GABA decreased the maximal velocity of the taurine uptake process and also decreased the Kt for taurine. Based on these data, it is proposed that: (i) there are two distinct transport systems, namely the GABA transporter and the taurine transporter, in these membranes which accept both GABA and taurine as substrates, (ii) the affinities of these systems for taurine are very similar and cannot be kinetically distinguished under the experimental conditions employed, and (iii) the difference between the affinities of these system for GABA is much greater than for taurine.

Amino Acids

Structure of the human pituitary adenylate cyclase activating polypeptide (PACAP) gene.

The human gene encoding pituitary adenylate cyclase activating polypeptide (PACAP) was isolated and its nucleotide sequence was determined. By comparison with a human PACAP cDNA, the exon/intron organization of PACAP gene was determined. The last exon encoded the longer form of PACAP, PACAP38 and 3'-untranslated sequences, suggesting that the shorter form of PACAP, PACAP27 is not generated by alternative splicing mechanisms. The 5'-flanking region of the PACAP gene contains several sequence motifs homologous to CRE, TRE, and GHF-1. On the basis of DNA isolated from mouse A9 microcell hybrid clone containing a single human chromosome, the PACAP gene was assigned to human chromosome 18. Furthermore, we determined the locus of the gene to be 18p11 by the chromosomal in situ hybridization technique.

Amino Acid Sequence

Antitumor effect of RBS (rice bran saccharide) on ENNG-induced carcinogenesis.

We examined whether orally administered RBS (rice bran saccharide), prepared from rice bran by hot water extraction, increases immunocompetence, inhibits gastrointestinal carcinogenesis with N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) or shows an antitumor effect. After the administration of RBS, phytohemagglutinin (PHA)- and pokeweed mitogen (PWM)-stimulated blastogenesis of lymphocytes derived from the mesenteric lymph nodes and peripheral blood was enhanced, and the helper/suppressor T-cell ratio was elevated, and migration activity of peritoneal macrophages was also increased in rats treated continuously with ENNG. ENNG-induced gastrointestinal carcinomas were observed in 43% of those administered RBS (ENNG-RBS) as compared with 88% in the control (ENNG) and 94% in the prednisolone (PRD) group (ENNG-PRD). The 12-month survival rate of rats bearing gastrointestinal cancer was 58% in the ENNG-RBS group as compared with 25% in the ENNG group and 15% in the ENNG-PRD group. RBS prevented the reduction in immunocompetence in the course of carcinogenesis, suppressed carcinogenesis, and prolonged the survival of rats with gastrointestinal cancer. Antitumor activities of RBS are thought to be a kind of host mediated action. The growth inhibition ratio of transplantable ENNG-induced cancer in Wistar rats was 42.1% in the RBS and 51.8% in the 5-FU group. Since little is known about the potent antitumor activity of alpha-glucan, it would be interesting to consider the relationship between the structure and the biological activities of polysaccharides.

Adjuvants, Immunologic

Cardiac function and myocardial performance of 24-hour-preserved asphyxiated canine hearts.

A method of 24-hour storage of asphyxiated canine hearts for orthotopic cardiac transplantation was studied to expand the geographical size of the donor pool. Left ventricular function of asphyxiated hearts preserved for 24 hours (group 1, n = 8) was compared with that of hearts donated on-site (group 2, n = 5). Group 1 donors were pretreated with verapamil hydrochloride, propranolol hydrochloride, and prostacyclin. The donor hearts were perfused with warm blood cardioplegia in situ after 10 minutes of asphyxiation and then perfused with cold crystalloid cardioplegia for 2 hours. The hearts were excised and stored in ice-cold University of Wisconsin solution for 22 hours. At orthotopic transplantation, coronary perfusion with warm blood cardioplegia was performed before the graft aorta was unclamped. Conventional cardiac variables (eg, cardiac output and maximum rate of rise of left ventricular pressure), myocardial performance, and diastolic compliance of grafted hearts were assessed 1 hour after weaning from bypass. All recipients in both groups were easily weaned from cardiopulmonary bypass without inotropic agents, and there were no significant differences in cardiac variables between the two groups. These results strongly suggest that cadaver hearts can be preserved for 24 hours with satisfactory cardiac function.

Animals

Transaortic patch angioplasty for left coronary ostial stenosis in a patient with Takayasu's aortitis.

A 35-year-old woman who had left coronary ostial stenosis and aortic valve regurgitation due to Takayasu's aortitis underwent transaortic patch enlargement of the stenosed left coronary ostium in combination with aortic valve replacement. This technique may be suitable and recommendable as an alternative to aortocoronary bypass grafting or endarterectomy for coronary ostial stenosis in Takayasu's aortitis.

Adult

Bactericidal activity of alkyl peroxyl radicals generated by heme-iron-catalyzed decomposition of organic peroxides.

To clarify the nature of cytocidal molecular species among the radicals generated in the iron-catalyzed reactions of peroxides (ROOH), we examined the cytocidal effects of these radicals against gram-positive and gram-negative bacteria in the presence or absence of various radical scavengers. Three organic peroxides, t-butyl hydroperoxide (t-BuOOH), methyl ethyl ketone peroxide (MEKOOH), and cumene hydroperoxide, were used. Each radical generated from these peroxides was identified and quantitated by electron spin resonance (ESR) spin trapping with 5,5-dimethyl-1-pyrroline-N-oxide (DMPO). The major cytotoxic radical species generated in the mixtures of various peroxides and heme iron, especially methemoglobin, metmyoglobin, or hemin, was the alkyl peroxyl radical (ROO.). Strong bactericidal action against gram-positive bacteria was observed in the peroxide-heme iron system, especially in the case of t-BuOOH and MEKOOH. Killing curves for gram-positive bacteria showed an initial lag period, which may indicate the multihit/multitarget kinetics of cell killing. When the diethylenetriamine pentaacetic acid (DTPA)-Fe2+ complex was used as a catalyst for decomposition of various peroxides, alkyl, alkoxyl, and alkyl peroxyl radicals were identified by spin-trapping analysis. However, study of the time course of alkyl peroxyl radical production in the DTPA-Fe2+ complex system revealed that radical species generated in this system were very short lived: a maximal level was achieved within 1 min and then declined sharply, and no bactericidal activity was observed after 10 min. In contrast, the alkyl peroxyl radical level generated by the organic peroxide-heme iron system remained high for 30 min or longer. The generation of alkyl peroxyl radicals quantified by ESR correlated quite well with the bactericidal effect of the system of peroxide plus iron. In addition, bactericidal activity was completely inhibited by the addition of the spin trap DMPO, as well as of other various radical scavengers (alpha-tocopherol and L-ascorbic acid), into the peroxide-heme iron system, but this effect was not observed with superoxide dismutase, beta-carotene, dimethyl sulfoxide, diphenylamine, or butylated hydroxyltoluene. In view of these results, it is assumed that alkyl peroxyl radicals are the potent molecular species that are cytotoxic against bacteria, whereas alkoxyl radicals (RO.) generated in this system do not affect bacterial viability.

Benzene Derivatives

Experimental evidence of selective axonal regeneration in allogenic and isogenic Y-chambers.

Studies on axonal regeneration in Y-chambers over the past decade have consistently provided evidence of preferential growth toward a distal nerve piece. However, these findings are in contrast to the observations by Weiss and Taylor (1944. J. Exp. Zool. 95: 233-257), indicating that in fresh aortic Y-chambers regenerating axons had no tendency to grow into a channel containing a distal nerve piece as compared to an open channel. The discrepancy between Weiss and Taylor's findings and those of later authors remains unexplained. In the present study we repeated the investigations by Weiss and Taylor, using both isogenic and allogenic aortic Y-chambers. In control groups the aorta graft was frozen and treated chemically to kill all cells and to deactivate possible growth factors of protein nature. In these groups preferential growth toward the distal nerve was pronounced although such specificity in growth was evident also in the nontreated types of aortic chambers. The findings do not support the results presented by Weiss and Taylor.

Animals

Bypass surgery for aortitis syndrome: aortocarotid bypass with saphenous vein graft.

In seven patients with Takayasu arteritis, symptomatic cerebral ischemia was successfully treated by extracranial bypass surgery. We performed 14 surgical procedures, which included aortocarotid bypass, axilloaxillary bypass, carotid-vertebral bypass, subclavian-carotid bypass, axillocarotid bypass, carotid-carotid bypass, carotid-subclavian bypass, carotid-axillary bypass, and thyreocervical-carotid anastomosis. Among those the aortocarotid bypass was the most common procedure performed--six occasions in four cases. The others were less common procedures, each performed only one time. The saphenous vein was used for the graft in 12 of the procedures. An artificial graft made of Gore-Tex (6-mm inner diameter) was utilized in two cases. Patency of the grafts was adequate and no mortality or serious postoperative complication was encountered. Asymptomatic infarction developed in one patient. We analyze these procedures in detail and stress the usefulness of aortocarotid bypass with a saphenous vein graft.

Adult

Relationships among the symptoms of the elderly as revealed by multivariate analysis: a preliminary study.

One hundred and sixty-eight elderly subjects in a geriatric hospital and the adjoining nursing home were examined in a study of relationships among the items of Hasegawa's dementia scale. A three-dimensional model using multivariate analysis demonstrated the relationships among items. The results led to the identification of six clinical groups determined by the presence of stroke and the score on the dementia scale. Nonstroke subjects of middle intellectual level had disturbances of memory and orientation. This group fell into two categories during the course of a five-year follow-up study: severe dementia and intellectual deterioration. Retrospective multivariate analysis results suggested that features characteristic of the severe intellectual deterioration group were earlier disturbances of orientation and distant memory. This statistical method uses only objective clinical data in order to avoid subjective and, often, biased judgments.

Aged

Constitutive expression of the taurine transporter in a human colon carcinoma cell line.

The human colon carcinoma cell line HT-29, when grown to confluence, was found to take up taurine and accumulate it against a concentration gradient from a NaCl-containing uptake medium. Replacement of NaCl with choline chloride almost totally abolished the uptake. Taurine uptake was dependent not only on Na+ but also on Cl-, because other anions failed to support the uptake in the presence of Na+. The uptake process was specific for beta-amino acids such as taurine, hypotaurine, and beta-alanine. Apparently, a single transport system with a Michaelis-Menten constant of 10.6 +/- 0.3 microM was responsible for the uptake. Stoichiometric analyses revealed that the Na+:taurine coupling ratio was 2:1, whereas the Cl-:taurine coupling ratio was 1:1. Culture of the cells in the presence of taurine caused downregulation of the uptake system. These cells were also capable of accumulating beta-alanine against a concentration gradient in the presence of NaCl. Beta-Alanine uptake occurred via a single transport system with an apparent Michaelis-Menten constant of 36 +/- 2 microM. Taurine and beta-alanine exhibited mutual interaction during uptake. Kinetic experiments strongly suggested that a common transporter was responsible for the uptake of these two beta-amino acids. It is concluded that the HT-29 cells constitutively express the taurine transporter and that this cell line may be a suitable model for investigations of intestinal taurine transporter.

Alanine

Dynamics of ventilation, circulation, and gas exchange to incremental and decremental ramp exercise.

Transient responses of minute volume (VE), O2 uptake (VO2), CO2 output (VCO2), heart rate (HR), and cardiac output (Q) to a step change and ramp changes with slopes ranging from 33.3 to 14.3 W/min were studied in five healthy human subjects over the load range from 25 to 125 W. The ramp responses were fitted to a first-order model with a pure time delay (td) and a time constant (TC), while most of the step responses fitted better to a second-order model, consisting of a fast and a slow component. No significant asymmetry was observed between the on- and off-responses to step forcing. The mean response time (MRT = td+TC) of the incremental ramp response was prolonged, whereas the MRT of the decremental ramp response was shortened or unchanged, with decreasing ramp slope. The asymmetry was commonly observed in respiratory and gas exchange variables and, to a lesser extent, also in circulatory variables. Neural and humoral factors that might be responsible for this phenomenon are discussed.

Adult

Primary adult T-cell lymphoma of the breast.

A case of primary adult T-cell lymphoma (ATL) of the breast is described. A 69-year-old woman presented with a painless, rapidly growing lump in her left breast. Staging procedures demonstrated no sign of generalized disease. Following a Patey's mastectomy, 10 courses of adjuvant chemotherapy (CHOP) were successfully administered. The light microscopic, immunohistochemical and molecular genetic analyses of the surgical specimen revealed a primary ATL. Seropositive mothers who breast-feed their children may facilitate the accumulation of T cells carrying HTLV-I in their breasts and thereby increase their risks of developing breast ATL.

Aged

Dynorphin-(1-13): antinociceptive action and its effects on morphine analgesia and acute tolerance.

Antinociceptive actions and effects of intracerebroventricular (i.c.v.) dynorphin-(1-13) (DYN) on morphine (MOR) analgesia and acute tolerance were studied in male Sprague-Dawley rats. Antinociceptive effect against hind paw pressure was produced by 30 micrograms of DYN, but not by 0.5-10 micrograms. Acetic acid writhing was inhibited dose-dependently by DYN at the doses of 2-30 micrograms, and the order of potency of the anti-writhing effect was beta-endorphin > MOR > DYN >> Met-enkephalin. The anti-writhing effect of DYN that was partially antagonized by naloxone at 10 mg/kg, s.c. in MOR tolerant rats was the same as that in MOR naive rats. The anti-writhing effect of i.c.v.-MOR was increased synergistically by DYN. Continuous s.c. (6 mg/kg/hr) and i.c.v. (7.5 micrograms/rat/hr) infusion of MOR produced antinociception against hind paw pressure, which reached maximum (MAX) and attenuated thereafter during MOR infusion for 6 hr. The attenuation of antinociception was also produced during MOR infusion combined with multiple i.c.v.-injection of DYN. The MAX and area under the antinociceptive curve during MOR infusion was not affected by multiple injection of DYN, i.e., no effect of i.c.v.-DYN on the development of acute MOR tolerance induced by s.c.- and i.c.v.-infusion was observed. In conclusion, the anti-writhing effect of i.c.v.-DYN might not be mediated via mu-receptors, although DYN increased the anti-writhing effect of i.c.v.-MOR synergistically and the development of acute tolerance to MOR (i.c.v., s.c.) was not affected by i.c.v.-DYN.

Analgesics

Time-related morphological changes of porcine ovarian granulosa cells incubated with urea-EDTA solution.

Morphological profiles of porcine granulosa cells incubated with 10 mM Tris-HCl containing 1 M urea and 5 mM EDTA (urea-EDTA solution) were investigated. The percentages of granulosa cells incorporating the dye, trypan blue on incubation with urea-EDTA solution did not change during the initial 30 min. Thereafter, granulosa cells gradually took up the dye throughout the incubation. The amount of protein released from granulosa cells increased dramatically during initial 15 min of incubation, but decreased during the following 15-30 min of incubation. Thereafter, the amount of proteins released from granulosa cells increased gradually again. The releasing profile of 51Cr-bounded substances in granulosa cells increased markedly during the initial 15 min of incubation, and decreased during the next 15 min of incubation. Subsequently the amount of 51Cr released was enhanced. The min of incubation. Subsequently the amount of 51Cr released was enhanced. The plasma membranes of granulosa cells remained intact at 30 min of incubation, although chromatin clusters of granulosa cells disappeared. Thereafter, a number of cells showed signs of degeneration, including broken plasma membrane and cytolysis. The present study revealed that urea-EDTA solution is useful in extracting materials from porcine granulosa cells. The majority of the materials extracted from granulosa cells during the initial 30 min of incubation with urea-EDTA solution is considered to be from the cell surfaces and/or intercellular matrix.

Animals

Behavior of cardiac output during progressive exercise tests: a preliminary report.

The behavior of cardiac output (Q) during progressive incremental exercise tests was studied in young healthy men. Q approached a plateau and leveled off at almost the same work rate at which oxygen uptake (VO2) attained its maxima, while heart rate (HR) still continued to rise. This suggests that the limiting factor for maximal aerobic capacity in healthy subjects is Q. The rate of increase in Q and HR accelerated from a work rate which is close to the ventilatory anaerobic threshold (AT). The prime cause of the progressive augmentation in cardiac activity is probably an accelerated release of plasma catecholamine and/or potassium. There is a possibility that these substances might also affect the AT.

Adolescent