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Biomedical subjects

Hiroyasu Satoh

Publications and source records attributed to Hiroyasu Satoh.

At least 19 recordsLinked to original sources

Analysis of polyhydroxyalkanoate (PHA) synthase gene in activated sludge that produces PHA containing 3-hydroxy-2-methylvalerate.

The identification of phaC which encodes PHA synthase, that is involved in the formation of polyhydroxyalkanoate (PHA) containing 3-hydroxy-2-methylvalerate (3H2MV), was attempted. As of now, PHA containing 3H2MV has been reported to be produced only by mixed microbial cultures in activated sludge, but no pure bacterial cultures. A laboratory-scale activated sludge process was operated for 67 days. During the operation of the activated sludge process, its capacity to produce PHA containing 3H2MV, and the diversity of the partial phaC genes in the activated sludge microorganisms were monitored periodically. Analysis of the partial phaC genes was conducted by PCR followed by cloning and DNA sequencing, or by PCR followed by terminal-restriction fragment length polymorphism (T-RFLP). The cloning-sequencing of the 263 clones gave 11 distinct genetic groups (GGs). All of the genetic groups had similarities to known phaC higher than 48%, and one of them had similarity as high as 96% to that of Alcaligenes sp. The behavior of each of the genetic groups during the operation of the activated sludge process was monitored by the T-RFLP method. The restriction enzyme AccII, with the help of MboI, enabled the monitoring of each of the genetic groups. One of the genetic groups was found to have a strong correlation with the capability of the activated sludge to produce PHA containing 3H2MV, and its DNA sequence together with its amino acid sequence are reported.

Acyltransferases↗

Abundance, diversity, and dynamics of viruses on microorganisms in activated sludge processes.

We examined the abundance of viruses on microorganisms in activated sludge and the dynamics of their community structure. Direct counting with epifluorescence microscopy and pulsed-field gel electrophoresis (PFGE) were applied to 20 samples from 14 full-scale wastewater treatment plants (wwtps) treating municipal, industrial, or animal wastewater. Furthermore, to observe the dynamics of viral community structure over time, a laboratory-scale sequencing batch reactor was operated for 58 days. The concentrations of virus particles in the wwtps, as quantified by epifluorescence microscopy, ranged from 4.2 x 10(7) to 3.0 x 10(9) mL-1. PFGE, improved by the introduction of a higher concentration of Tris-EDTA buffer in the DNA extraction step, was successfully used to profile DNA viruses in the activated sludge. Most of the samples from different wwtps commonly had bands in the 40-70 kb range. In the monitoring of viral DNA size distribution in the laboratory-scale reactor, some bands were observed stably throughout the experimental period, some emerged during the operation, and others disappeared. Rapid emergence and disappearance of two intense bands within 6 days was observed. Our data suggest that viruses--especially those associated with microorganisms--are abundant and show dynamic behavior in activated sludge.

Animals↗

Local regulator adrenomedullin contributes to the circulatory disturbance in cirrhotic rats.

AIM: To investigate whether adrenomedullin, a potent vasodilator peptide, plays a role in the circulatory disturbance in cirrhosis. METHODS: Cirrhosis was induced in rats by weekly gavage of carbon tetrachloride. Hemodynamic studies were performed in vivo using radioactive microspheres and in vitro using isolated aortic rings. The adrenomedullin concentrations were measured by radioimmunoassay. RESULTS: Acute administration of adrenomedullin to the control rats reduced the systemic arterial pressure along with an increase of serum levels of the stable metabolite of nitric oxide (NOx), in a dose-dependent manner. Chronic infusion of adrenomedullin reduced the vascular resistance and increased the blood flow in the systemic and splanchnic circulation. Intravenous administration of anti-adrenomedullin antibody did not affect any hemodynamic parameters in the cirrhotic rats, whereas this antibody ameliorated the blunted contractile response to phenylephrine, alpha-adrenergic receptor agonist, in the aortic rings of the cirrhotic rats. The adrenomedullin concentrations in the aorta were higher in the cirrhotic rats than in the controls, and correlated with the mean arterial pressure in the cirrhotic rats. Moreover, adrenomedullin blunted the contractile response to phenylephrine in both of the control aorta and cirrhotic aorta, but not in the presence of NG-nitro-L-arginine methyl ester, an NO synthase inhibitor. CONCLUSION: Adrenomedullin overproduced in the vascular wall may contribute to the circulatory disturbance in cirrhosis as a local regulator of the vascular tonus rather than a circulating hormone.

Adrenomedullin↗

Effect of soluble microbial products on microbial metabolisms related to nutrient removal.

In this study, the effect of soluble microbial products (SMP) on the metabolisms related to phosphate or nitrogen removal of activated sludge was investigated. Two anaerobic-aerobic activated sludge processes were operated, one with a hydraulic retention time (HRT) of 48 h (RunL) and the other 6.4 h (RunS). The longer HRT of RunL was intended to promote the accumulation of SMPs in the supernatant. With the sludge from RunS and the supernatant from both of the runs, supernatant exchange batch experiments (SEBEs) were conducted, in which the acetate uptake rate and phosphate release rates under anaerobic conditions and the phosphate uptake rate under aerobic conditions were measured as these metabolisms are related to enhanced biological phosphorus removal. The nitrification rate was also measured. The statistical analyses of the results from the SEBEs showed that the supernatant from RunL had an inhibitory effect on the anaerobic acetate uptake and nitrification of the sludge from RunS. The cause of which was attributed to SMPs in the supernatant from RunL. As a result, the inhibitory effect of SMPs on nitrification and anaerobic acetate uptake was confirmed.

Sewage↗

In vitro pharmacological actions of sinomenine on the smooth muscle and the endothelial cell activity in rat aorta.

Vasodilating actions of sinomenine were examined using rat aorta ring strips. Sinomenine (0.1 to 100 microM) dilated norepinephrine (NE, 5 microM)-induced vasoconstriction in a concentration-dependent manner reaching 68.8+/-5.1% (n=6, P<0.01) at a concentration of 100 microM. Sinomenine (100 microM) also attenuated KCl (60 mM) and phorbol 12, 13-dibutyrate (PDB, a protein kinase C, PK-C, activator, 300 nM)-induced vasoconstriction by 86.9+/-8.5% (n=6, P<0.01) and 49.9+/-9.8% (n=6, P<0.01), respectively. Pretreatment with nicardipine (a Ca2+ channel antagonist), staurosporine (a PK-C inhibitor), NG-monomethyl-L-arginine acetate (L-NMMA, a nitric oxide, NO, synthesis inhibitor), and indomethacin (a cyclooxygenase inhibitor) were carried out. Nicardipine (0.1 microM) led to a significant decrease in the vasodilating potential of sinomenine (at 100 microM, 68.8+/-5.1% vs. 35.5+/-6.9%; n=5, P<0.001). Pretreatment with staurosporine (30 nM) reduced sinomenine-associated vasodilation from 68.8+/-5.1% to 49.5+/-7.7% (n=5, P<0.001), and L-NMMA (100 microM) and indomethacin (10 microM), to 25.3+/-2.3% (n=5, P<0.001) and to 37.1+/-9.3% (n=5, P<0.001), respectively. The responses were almost similar to the results without endothelium. Therefore, these results indicate that sinomenine causes the vasorelaxation by the mechanisms involved with the inhibitions of Ca2+ channel and PK-C activity, and also with the activations of NO and prostaglandin (PG) I2 syntheses in endothelium.

Animals↗

Decreases of calcium and phosphorus in monkey cardiac walls with development and aging.

To elucidate compositional changes of the cardiac walls with development and aging, the authors investigated changes of elements in the atrial and ventricular walls of monkeys. The left and right atrial walls, left and right ventricular walls, and interatrial and interventricular septa were resected from the subjects. The subjects consisted of 17 rhesus and 13 Japanese monkeys, ranging in age from 10 d to 33 yr. The element content of the cardiac walls was analyzed by inductively coupled plasma-atomic emission spectrometry. The Ca and P contents decreased in all of the left and right atrial and ventricular walls, interatrial septa, and interventricular septa with development, whereas the S and Mg contents decreased in the left and right ventricular walls with development. Regarding the relationships among elements, significant direct correlations were found among Ca, P, Mg, and Zn in all of the left and right atrial walls, left and right ventricular walls, and interatrial and interventricular septa, with some exceptions. As Ca decreased in the cardic walls, P, Mg, and Zn decreased simultaneously in the cardiac walls. The mass ratio of Ca/P decreased gradually with Ca decrease in both the atrial and ventricular walls, but it was not constant.

Aging↗

Subtypes of neuronal nicotinic acetylcholine receptors involved in nicotine-induced phosphorylation of extracellular signal-regulated protein kinase in PC12h cells.

Although many kinds of nicotinic acetylcholine receptor (nAChR) subtypes have been reported in the neuronal tissues, subtype differences in the nAChR-mediated intracellular signaling remains obscure. Using nAChR agonists and antagonists, the involvement of nAChRs in extracellular signal-regulated protein kinase (ERK) phosphorylation in PC12h cells was investigated. Cytisine and nicotine induced the phosphorylation of ERKs in a dose-dependent manner, whereas RJR-2403 had no effect. Cytisine, but not RJR-2403, also induced phosphorylation of CREB. Mecamylamine, dextromethorphan and 18-methoxycoronaridine inhibited nicotine-induced ERK phosphorylation with much higher affinity than dihydro-beta-erythroidine and alpha-conotoxin MII. These results suggest the involvement of alpha3beta4 nAChRs in ERK phosphorylation in PC12h cells.

Alkaloids↗

Suppression of pacemaker activity by Ginkgo biloba extract and its main constituent, bilobalide in rat sino-atrial nodal cells.

Effects of Ginkgo biloba extract (GBE) and bilobalide (a main constituent) on the pacemaker activity and the underlying ionic currents in rat sino-atrial (SA) nodal cells were investigated using patch-clamp techniques. Both GBE and bilobalide depressed the pacemaker activity in a concentration-dependent manner. At both 0.03 mg/ml GBE and 0.3 microM bilobalide, a negative chronotropic effect was produced. Dysrhythmias often occurred. The L-type Ca(2+) current (I(Ca)) and the hyperpolarization-activated inward current (I(f)) decreased by 69.7+/-3.2% (n=6, P<0.001) and by 12.6+/-2.1% (n=7, P<0.05) at 0.03 mg/ml GBE, and by 51.2+/-3.3% (n=6, P<0.01) and by 19.8+/-2.2 % (n=6, P<0.05) at 0.3 microM bilobalide, respectively. The delayed rectifier K(+) current (I(K)) also decreased. The inhibition was 12.3+/-2.0% (n=6, P<0.05) at 0.03 mg/ml GBE, and was 28.0+/-2.9% (n=6, P<0.05) at 0.3 microM bilobalide. These results indicate that cardiac ionic channels contributing to the pacemaking are highly sensitive to GBE and bilobalide, which can sufficiently modify the spontaneous activity in rat SA nodal cells.

Action Potentials↗

Age-related changes in the vasodilating actions of Ginkgo biloba extract and its main constituent, bilobalide, in rat aorta.

BACKGROUND: Age-related modulation in vasodilating actions induced by Ginkgo biloba extract (GBE) and bilobalide, a main constituent of GBE, were examined using rat aorta ring strips. METHODS: Wistar rats from 5 to 25 weeks old were used, and the isolated aorta ring strips were fixed in Krebs-Henseleit solution. RESULTS: GBE and bilobalide concentration-dependently dilated norepinephrine (NE)-induced vasoconstriction in all aged rats. The vasodilating actions generally decreased in accordance with aging. GBE at 1 mg/ml decreased from 28.4+/-3.8% (n=5) in 5-week-old rats to 23.7+/-7.1 (n=7) in 25-week-old rats, but not significantly. GBE (3 mg/ml)-induced vasodilation was maximum by 73.7+/-2.1% (n=4, P<0.001) in 10-week-old rats. GBE had the marked vasodilation at younger ages and further decreased it with developing ages. In the rats older than 20 weeks, however, GBE tended to rather increase the strength of vasodilating action. On the other hand, the vasorelaxation induced by 30 micromol/l bilobalide significantly decreased from 11.8+/-1.4% (n=4) in 5-week-old rats to 2.3+/-1.5% (n=5, P<0.01) in 25-week-old rats, and by 100 micromol/l from 20.2+/-3.4% (n=4) to 5.6+/-2.5% (n=5, P<0.01), respectively. Bilobalide had the similar age-related actions. The age-dependent attenuation was produced milder by bilobalide than by GBE. At lower concentrations, however, bilobalide caused the weak vasocontriction in 20- and 25-week-old rats. CONCLUSION: GBE and bilobalide possess a similar characteristic for age-related modification, clinically suggesting the more effective actions of GBE for elder persons.

Age Factors↗

Electropharmacological actions of the constituents of Sinomeni Caulis et Rhizome and Mokuboi-to in guinea pig heart.

Cardiac actions of the constituents of Sinomeni Caulis et Rhizome (SCR) and Mokuboi-to, a traditional herbal drug, were investigated. In voltage-clamp experiments, sinomenine (1 mM) and tetrandrine (100 microM) inhibited the ionic currents concentration-dependently. The constituents affected the action potential configurations. In multicellular preparations, SCR (1 mg/ml) decreased the maximum rate of depolarization. Tetrandrine (30 microM) and sinomenine (300 microM) also had similar effects, but magnoflorine (1 mM) had less or no effect. Dysrhythmias were abolished under Ca(2+) overload conditions by sinomenine. These results indicate that at even acute administrations, these drugs exert the active electropharmacological actions and cardioprotections.

Action Potentials↗

Age-dependent decreases of calcium, phosphorus, sulfur, and zinc in the cardiac valves of monkeys.

To elucidate compositional changes of the cardiac valves in monkey with aging, the authors investigated age-related changes of elements in the four cardiac valves of rhesus and Japanese monkeys and the relationships among elements by inductively coupled plasma-atomic emission spectrometry (ICP-AES). The four cardiac valves of the aortic, pulmonary, mitral, and tricuspid valves were resected from 19 rhesus and 11 Japanese monkeys, ranging in age from 10 d to 33 yr. The element contents were determined by ICP-AES. It was found that the Ca, P, S, and Zn contents were high in the four cardiac valves of the monkeys below 1 yr and thereafter decreased significantly with aging, except for Ca in the mitral valve, for which no significant correlation was found between age and Ca content. The Ca and P contents did not increase in the four cardiac valves at old age. This result revealed that calcification scarcely occurred in the four cardiac valves at old age. With regard to the relationships among elements, it was found that there were significant direct correlations among the Ca, P, S, and Zn contents in all of the four cardiac valves of the monkeys, with two exceptions between P and Zn contents in both the aortic and pulmonary valves. Therefore, as Ca decreased in the four cardiac valves, P, S, and Zn decreased simultaneously in the same cardiac valves.

Aging↗

Age-related attenuation in the elements in monkey sino-atrial node.

Changes in trace elements of the sino-atrial (SA) node with aging was investigated using 24 hearts of the Japanese and rhesus monkeys of ages ranging from 27 d to 30 yr. With aging, sympathetic activity decreases and SA nodal function deteriorates. The SA nodal tissue was removed from the anatomical position and was confirmed by means of histological observation. The elements, such as Ca, P, S, Mg, Na, Fe, and Zn, were analyzed using inductively coupled plasma-atomic emission spectrometry (ICP-AES). Advancing age never increased the contents of the trace elements, but decreased them. The correlation coefficients for the age-dependent attenuations were -0.561 (n = 24, p < 0.01) in Ca and -0.482 (n = 24, p < 0.05) in P. The correlations for the attenuations induced by other trace elements were not significant. Furthermore, close relationships of the elements between Ca and P, S, Zn, or Na contents, between P and Zn or Na contents, and between Zn and Na contents were observed. These results indicate that the elements in the monkey SA node are attenuated with an increase in age, presumably suggesting the age-related suppression of cardiac functions as a result of the histological alterations of the SA nodal cells.

Aging↗

Comparative Electropharmacological Actions of Some Constituents from Ginkgo biloba Extract in Guinea-pig Ventricular Cardiomyocytes.

Effects of the constituents from Ginkgo biloba extract (GBE) on the action potentials and the ionic currents in guinea pig ventricular cardiomyocytes were investigated using whole-cell and current-clamp techniques. The constituents, ginkgolides A, B, C and quercetin, had depressant effects at 0.1-3muM on the action potential configuration. Ginkgolide A (1-3 muM) prolonged the action potential (action potential duration: APD) at 75% and 90% repolarizations (APD(75) and APD(90)). However, ginkgolides B and C at low concentrations prolonged APD, but at higher concentrations (>1 muM) shortened APD. Quercetin at 3 muM prolonged the APD, but not at the lower concentrations. These constituents also inhibited the V(max). The resting potential was unaffected. In voltage-clamp experiments, ginkgolides A and B (0.1-3 muM) markedly and concentration-dependently increased the Ca(2+) current (I(Ca)) and the delayed rectifier K(+) current (I(K)), and decreased the inwardly rectifying K(+) current (I(K1)). On the other hand, ginkgolide C failed to affect the I(Ca) but increased the I(K) by 14.0 +/- 2.3% (n = 6, P < 0.05) at 1 muM. Quercetin inhibited I(Ca), and enhanced I(K) but decreased I(K1). These responses to the constituents were almost reversible (80-90% of control) after a 10- to 20-min washout. These results indicate that even at acute administrations, these constituents produce the effective actions on the APD and the underlying ionic currents in cardiomyocytes. Each constituent does not exhibit a uniform response, although GBE acts as a net.

Journal Article↗

Comparative vasodilating actions among terpenoids and flavonoids contained in Ginkgo biloba extract.

BACKGROUND: Comparative vasodilating actions of the constituents of Ginkgo biloba extract (GBE), terpenoids (bilobalide, ginkgolides A, B and C) and flavonoids (quercetin and rutin), were examined using rat aorta ring strips. METHODS: Cumulative administrations of GBE and its constituents were followed with the pretreatment of 5 micromol/l NE. RESULTS: GBE at 0.03 to 3 mg/ml had a potent concentration-dependent relaxation; by 70 +/- 4.5% (n = 6, P < 0.001) at 3 mg/ml. Terpenoids and flavonoids at 0.1 to 100 micromol/l had potent concentration-dependent relaxation. At 100 micromol/l, bilobalide dilated by 17.6 +/- 3.9% (n = 7, P < 0.05), and ginkgolides A, B and C also caused it to the almost same extent. Quercetin (100 micromol/l) caused a potent vasorelaxation by 49.9 +/- 4.8% (n = 10, P < 0.001). Rutin at 100 micromol/l had weaker vasorelaxation; by 13.7 +/- 3.2% (n = 6, P < 0.01). CONCLUSIONS: All constituents of GBE have the concentration-dependent vasorelaxtant effect. The potency of GBE's action was not made simply by addition of those of the constituents. Each constituent itself would contribute to the GBE-induced vasodilation, although the constituents have the complicated interactions with each other.

Animals↗

Electropharmacological actions of Ginkgo biloba extract on vascular smooth and heart muscles.

Ginkgo biloba extract (GBE) is composed mostly of two constituents: One is terpenoids (such as bilobalide, ginkgolides A, B and C), and the other is flavonoids (such as quercetin and rutin). After oral administration of GBE (160 mg) to healthy volunteers, the plasma concentrations of ginkgolides A and B and bilobalide are 41.8, 5.6 and 37.6 ng/ml, respectively. GBE and bilobalide cause a potent concentration-dependent relaxation. NG-Monomethyl-l-arginine acetate (l-NMMA), an NO synthesis inhibitor, reduces the vasodilation induced by GBE. Furthermore, the vasorelaxation of GBE is attenuated in Ca2+-free medium. Bilobalide possesses similar mechanisms. The other constituents also produce vasorelaxation. On the other hand, all the compounds markedly modify the action potential configuration in guinea pig ventricular cardiomyocytes. GBE prolongs the action potential duration (APD), whereas bilobalide shortens the APD. In patch-clamp experiments, GBE markedly inhibits the Ca2+ current (ICa), the delayed rectifier K+ current (IK) and the inwardly rectifying K+ current (IK1). On the contrary bilobalide enhances the ICa and IK currents concentration-dependently. The other constituents do not cause their actions in a uniform direction. In the rat sino-atrial (SA) node, GBE causes a negative chronotropic effect. These results indicate that GBE and the constituents produce effective electropharmacological actions in the cardiomyocytes and cause vasodilation, mainly due to the inhibitions of Ca2+ influx through the Ca2+ channel and the activation of NO release in the endothelium and aortic vascular muscles.

Action Potentials↗

Effects of substrate loading rate on biofilm structure.

The effects of substrate surface loading rate on biofilm growth and structure were investigated by chemical, biochemical and microscopic methods. Three tubular reactors were operated at equal C:N ratio of 0.1, with substrate loading rates of 1.2, 0.6 and 0.3g-C/m(2)/day. Substrate loading positively influenced the biofilm growth rate. Denser biofilms with lower porosities were formed at higher substrate loading. Slowly growing biofilms having porous structures were found to have higher specific activities. Nitrification was suppressed under the higher substrate loading conditions even at the equal C:N ratio of 0.1, thus proving that the spatial competition between nitrifiers and heterotrophs as one limiting criteria for stable nitrification. The spatial organization of the ammonia oxidizers was biofilm structure related. The strain variability of ammonia oxidizers was substrate loading dependent. These findings suggest that substrate loading is a key parameter in determining biofilm structure and function.

Ammonia↗

Mechanisms for the vasodilations induced by Ginkgo biloba extract and its main constituent, bilobalide, in rat aorta.

Vasodilating actions of Ginkgo biloba extract (GBE) and bilobalide, a main constituent, were examined using rat aorta ring strips. GBE at the concentration ranges from 0.03 to 3 mg/ml had a potent concentration-dependent relaxation, reaching 70 +/- 4.5% (n = 6, P < 0.001) at 3 mg/ml. Bilobalide at 0.1 to 100 microM also caused the relaxation in a concentration-dependent manner. At 100 microM, bilobalide caused dilation by 17.6 +/- 3.9% (n = 7, P < 0.05). NG-monomethyl-L-arginine acetate (L-NMMA)(100 microM), an NO synthesis inhibitor, reduced the vasodilation of GBE (3 mg/ml) to 57.6 +/- 2.5% (n = 6, P < 0.05), and was accompanied with a decrease in the rate of relaxation. Tetraethylammonium (TEA)(100 microM), a Ca(2+)-activated K(+) channel inhibitor, also decreased the GBE (3 mg/ml)-induced relaxation to 63.1 +/- 4.6% (n = 6), but not significantly. Indomethacin tended to reduce the GBE (3 mg/ml)-induced vasorelaxation to 67.3 +/- 4.1% (n = 6). In contrast, the vasorelaxation of GBE (3 mg/ml) was strongly attenuated to 53 +/- 6.1% (n = 7, P < 0.05) in Ca(2+)-free medium. Similarly, the vasorelaxation induced by bilobalide significantly decreased both by pretreatment with NO inhibitor (L-NMMA) and in Ca(2+)-free solution. These results indicate that the relaxation induced by GBE would be due to the inhibition of Ca(2+) influx through the Ca(2+) channel and the activation of NO release, and might be in part due to the inhibitions of Ca(2+)-activated K(+) current and PGI(2) release, in the endothelium and aortic vascular muscles. Bilobalide possesses the similar mechanisms for the vasodilation.

Animals↗