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H Watari

Publications and source records attributed to H Watari.

At least 55 records · Page 3Linked to original sources
PubMed1988

An experimental approach to 31P spin-lattice relaxation time measurement in biological systems.

The accuracy of measurement of the spin-lattice relaxation time (T1) in biological systems using the variable nutation angle method is discussed using computer simulations. This method could reduce the time required for measurement to 10% of that using the inversion recovery method. In the perfused salivary gland of the rat (310 K), the T1 values of sugar phosphates, creatine phosphate, gamma-phosphate, and beta-phosphate of nucleotide triphosphates determined by this method were 5.2, 4.8, 0.9, and 0.8 s, respectively. During secretion of saliva, the T1 value of sugar phosphates decreased to 3.7 s, which could be due to an increased chemical exchange of phosphate in glycolysis.

Animals↗
PubMed1988

Internal motion of a tryptophan residue in Streptomyces subtilisin inhibitor: deuterium nuclear magnetic resonance in solution.

Deuterium NMR spectroscopy was used to study internal motions of a deuterium-labeled single tryptophan (Trp) residue (per subunit) of Streptomyces subtilisin inhibitor (SSI) in solution. The free inhibitor with the five ring protons of the Trp replaced with deuterons showed a narrow resonance component (56 Hz) of about one-quarter of the total intensity, in addition to the broad resonance component (about 600 Hz) at 25 degrees C, showing that it exits in an equilibrium mixture of two conformers, in one of which the tryptophan side chain is highly mobile. In analogy to the two structures of SSI found in the crystal, these two conformers were attributed to the one in which the contact between the alpha-lobe and the beta-lobe of the subunit is tight and the other in which the same contact is loose. When SSI forms a complex with subtilisin BPN', the broad component becomes invisibly broad, but the narrow component increases with even further narrowing, suggesting that the binding to the enzyme favors the "loose" conformer over the "tight" conformer.

Bacterial Proteins↗
PubMed1988

NMR-invisible ATP in rat heart and its change in ischemia.

The subcellular compartmentalization of adenosine 5'-triphosphate (ATP) in isolated perfused rat heart and its relation to energy depletion in ischemia were examined by 31P nuclear magnetic resonance (31P-NMR) spectroscopy and chemical analyses. The signal intensities of the beta-phosphate of ATP and creatine phosphate in the 31P-NMR were standardized by the intracellular volume ratio measured with 23Na-NMR to determine the actual content of each. During aerobic perfusion the ATP content determined by NMR (13.7 +/- 2.2 mumol/g dry weight) was significantly lower than that found by chemical analysis (22.4 +/- 0.7 mumol/g dry weight), while the creatine phosphate contents determined by the two methods were the same. During ischemia at 33 degrees C, the signal of the beta-phosphate of ATP in the 31P-NMR spectrum decreased progressively, disappearing completely after 16 min. But at this time 5.7 +/- 1.7 mumol/g dry weight of myocardial ATP was still detected by chemical analysis. These results indicated that there were two different compartments of intracellular ATP in the heart, only one of which is detectable by 31P-NMR spectroscopy, and that during ischemia the ATP that is detectable, which seems to be the free ATP in the cytosol, decreased more rapidly than the ATP in the other compartment.

Adenosine Triphosphate↗
PubMed1988

Phosphorous compounds studied by 31P nuclear magnetic resonance spectroscopy in the taenia of guinea-pig caecum.

1. In the isolated taenia (0.4-0.6 g) of guinea-pig caecum, the intracellular phosphorous compounds and pH were investigated using 31P nuclear magnetic resonance (NMR) under various metabolic conditions. 2. The ratios of the intracellular concentration of phosphocreatine ([ PCr]) and inorganic phosphate [( Pi]) to nucleotide triphosphate ([ NTP]) were 1.71 +/- 0.14 and 0.58 +/- 0.11 (n = 25), respectively, in normal solution (32 degrees C). The intracellular pH estimated from the chemical shift of Pi was 7.05 +/- 0.06 (n = 25), agreeing well with those previously obtained. 3. In the absence of glucose, the [PCr] and [NTP] were decreased to almost a half after 150 min exposure to 40 mM-K+ solution, while [Pi] was increased 3-fold. These changes were much faster than the rate of decline in tension. When glucose was readmitted, the contractile response to K+ fully recovered in 50 min. However, this was accompanied with only a partial recovery of [PCr] and [Pi], but no recovery of [NTP]. The intracellular pH was lowered by about 0.2 of a unit, suggesting an increase in glycolysis. 4. In Ca2+-free solution, respiratory inhibition with hypoxia or CN (1 mM) only decreased [PCr], leaving [NTP] nearly unchanged. On the other hand, respiratory inhibition in excess-K+ solution containing Ca2+ (2.4 mM) severely depleted PCr and decreased [NTP] to 40%. Increasing glucose to 50 mM did not prevent these changes, although it increased tension development. 5. The simultaneous decrease of [NTP] and [PCr] during K+ contracture suggests that the activity of creatine phosphokinase is low. The recovery from respiratory inhibition was much better for [PCr] than for [NTP]. Slow, but perfect, recovery of all NTP peaks was produced by adding 1 mM-adenosine to normal solution. 6. It was suggested that tension development is closely related to the turnover rate of ATP, and not to its concentration, and that deamination of adenosine is a limiting factor in the recovery of ATP after excessive consumption.

Adenosine↗
PubMed1988

Phosphorus nuclear magnetic resonance in isolated perfused rat pancreas.

Phosphorus nuclear magnetic resonance spectroscopy was applied to measure phosphorus energy metabolites in isolated perfused rat pancreas. The gland was perfused with a modified Krebs-Henseleit solution at room temperature (25 degrees C). 31P resonances of creatine phosphate (PCr), ATP, ADP, inorganic phosphate (Pi), and phosphomonoesters (PMEs) were observed in all the preparations of pancreas. In different individual preparations, the resonance of PCr varied, but those of ATP were almost the same. The initial levels of PCr and ATP in individual preparations, however, remained almost unchanged during perfusion with the standard solution for 2 h. When the perfusion was stopped, the levels of ATP and PCr decreased, while the levels of PME and Pi increased. At that time, the Pi resonance shifted to a higher magnetic field, indicating that the tissue pH decreased. On reperfusion, the tissue levels of phosphorus compounds and the tissue pH were restored to their initial resting levels. Continuous infusion of 0.1 microM acetylcholine caused marked and sustained increases in the flow of pancreatic juice and protein output. During the stimulation the tissue levels of phosphorus compounds remained unchanged, while the tissue pH was decreased slightly (0.05 pH unit).

Adenine Nucleotides↗
PubMed1988

Dissociation of fluid secretion and energy supply in rat mandibular gland by high dose of ACh.

The effects of a high dose of acetylcholine (ACh) on oxygen consumption (VO2) and changes in phosphorus energy metabolites during secretion were studied in isolated perfused mandibular gland of rats at 24 degrees C. Sugar phosphates (SP), Pi, phosphocreatine (PCr), and ATP were identified by phosphorus-31 nuclear magnetic resonance spectroscopy. One micromole ACh induced a tachyphylactic secretory response, a persistently elevated VO2, and decreased PCr and ATP; 1 mM ACh caused an initial burst of secretion that was followed by suppression of secretion and a rapid increase in the VO2 to the same level as that with 1 microM ACh. These findings indicate a dissociation between secretion and VO2. During stimulation with 1 mM ACh, the level of PCr first decreased and then partially recovered, but the level of ATP continued to decrease and the levels of Pi and SP increased markedly. These findings suggest compartmentalization of creatine phosphokinase (CPK) systems and the possibility that a high concentration of ACh interferes with the transport of PCr between one CPK system near adenosinetriphosphatase and another system near mitochondria in acinar cells.

Acetylcholine↗
PubMed1988

In vivo 31P MRS in new antineoplastic agents evaluation on experimental tumor models.

Two new antineoplastic agents, a nitrosourea and a DNA-bis-intercalator have been studied in vivo by 31P magnetic resonance spectroscopy on a rat glioma and Walker carcinoma. On rat glioma, spectra are modified when the tumor is treated by the nitrosourea, showing the depletion of high-energy phosphates. On Walker carcinoma both drugs delay the tumor evolution to necrosis, showing important levels of high-energy phosphates on NMR spectra. There appears to be a great dependence upon energy metabolism during chemotherapy, depending on the nature and physiology of the observed tumor.

Animals↗
PubMed1987

Direct measurement of Na influx by 23Na NMR during secretion with acetylcholine in perfused rat mandibular gland.

Intracellular Na content (Nain) in the perfused rat mandibular gland was measured by using a 23Na NMR spectroscopy at 24 degrees C. An aqueous chemical shift reagent, dysprosium triethylenetetramine-N,N,N',N",N"'N"'-hexaacetic acid [Dy(TTHA)] was used in order to discriminate between the intracellular and the extracellular Na signal. The mandibular gland of rat was perfused arterially with a modified Krebs solution containing 10 mM Dy(TTHA). At rest, Nain was not changed by blocking the Na+/K+ ATPase with ouabain (1 mM) and atropine (3 microM), implying that, in the absence of stimulation, the spontaneous Na influx across the plasma membrane must have been negligibly small. Following onset of stimulation with acetylcholine (1 microM), Nain increased by 9.1 +/- 1.5 mmol/l intracellular fluid (mean +/- SEM, n = 13), and remained at this level during stimulation. In the initial phase of secretion (0-5 min), about 50 mmol/min/l intracellular fluid of Na was secreted into the luminal space (estimated from the secretory rate by assuming an isotonic primary secretion) but, in spite of the higher secretion rate, Nain increased only at an initial rate of 4.1 mmol/min/l intracellular fluid. During the steady phase of secretion (15-30 min) evoked by acetylcholine (1 microM), ouabain (1 mM) caused an increment of Nain of 44 +/- 8 mmol/l intracellular fluid (mean +/- SEM, n = 4). From the rate of Nain increment, the Na influx rate at the steady phase was estimated as 4.5 mmol/min/l intracellular fluid. These results suggest that the influx of Na is caused by stimulation with acetylcholine.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1987

Multiple forms of immunoreactive renin in human adrenocortical tumour tissue from patients with primary aldosteronism.

There is increasing evidence which suggests that the adrenal gland contains the renin-angiotensin cycle. The localization of renin has been reported to be mainly in the zona glomerulosa rather than the fasciculata medullary portion. In the present study we have investigated extracts from aldosteronomas (n = 3), which are believed to derive from the zona glomerulosa cells. In addition, we have attempted to characterize the biochemical properties of the adrenal renin. Sizable quantities of renin-like activity (32.0 +/- 7.7 ng of angiotensin I generated h-1 mg-1 of protein, mean +/- SEM) were detected in the extracts. This renin-like activity was inhibited by anti-renin antibody raised against pure renin (mean, 95% of the total renin-like activity), indicating that it was not due to the non-specific action of proteases such as cathepsin D. The optimum pH of the tissue renin-like enzyme was 6.0 for rat plasma substrate. Differences were found, however, in the molecular mass (36,000, 37,000, 44,000 and 48,000), binding to concanavalin A and isoelectric points (4.40, 4.68 and 5.00). These results confirm the existence of specific renin in aldosteronoma. Renin microheterogeneity could be evidence for local production of the enzyme.

Adrenal Cortex Neoplasms↗
PubMed1987

High resolution 23Na-nuclear magnetic resonance study of stroke-prone spontaneously hypertensive rat erythrocytes.

The intracellular Na+ content of washed erythrocytes from stroke-prone spontaneously hypertensive rats (SHRSP) and Wistar-Kyoto normotensive rats (WKY) was measured by a high resolution 23Na-nuclear magnetic resonance (NMR) technique using a non-permeant aqueous shift reagent, dysprosium triethylenetetramine hexaacetic acid, Dy(TTHA)3-. The initial intracellular Na+ of freshly isolated and washed erythrocytes was very low (approximately 5 mmol/l) and increased progressively with prolonged incubation in isotonic salt solution at 37 degrees C. There was no significant difference in the erythrocyte Na+ concentration between SHRSP and WKY over the entire period of measurement, nor was any difference detected in their osmotic fragility or total cellular volume, although the osmotic fragility decreased with incubation time. The high energy phosphate metabolites were also studied in the same erythrocytes by 31P-NMR. The level of intracellular ATP decreased with incubation at 37 degrees C but showed no difference between the SHRSP and WKY samples. Inclusion of 1 mmol/l ouabain in the incubation medium substantially retarded the breakdown of intracellular ATP and resulted in a concomitant increase in intracellular Na+. However, neither the ouabain-sensitive nor the ouabain-insensitive component of Na+ influx altered in SHRSP erythrocytes compared with WKY erythrocytes in paired experiments. Our results do not support the hypothesis that altered Na+ transport, resulting in an increase in erythrocyte Na+ concentration, is associated with spontaneous hypertension.

Animals↗
PubMed1987

31P NMR studies on the isolated perfused mandibular gland of the rat.

Phosphorus nuclear magnetic resonance (31P NMR) was used to study energy metabolism in the rat mandibular gland. The gland was isolated, perfused arterially and set in the NMR tube. At rest, 7 resonance peaks were observed and 6 peaks identified from low field as: 1) sugar phosphates (SP) and nucleotide monophosphate (NMP), 2) inorganic phosphate (Pi), 3) creatine phosphate (PCr), 4) gamma-nucleotide triphosphate (NTP) and beta-nucleotide diphosphate (NDP), 5) alpha-NTP, alpha-NDP, NAD+, and NADH, 6) an unknown peak, and 7) beta-NTP. From the results of high performance liquid chromatography (HPLC), NTP consisted mainly of ATP and GTP, and UTP was not detected. The tissue contents of ATP and GTP in the perfused gland were determined by HPLC as 1.86 +/- 0.03 and 0.37 +/- 0.01 mmol/kg wet tissue (S.E., n = 5). From 31P NMR and HPLC data, the tissue levels of creatine phosphate, ADP, and sugar phosphates were estimated as 3.3, 0.4, and 4.2 mmol/kg wet tissue, respectively. The cessation of perfusion decreased the tissue levels of PCr and ATP and increased those of Pi and SP. On the other hand, administration of acetylcholine (1 microM), which is an optimal dose for secretion, decreased PCr and increased Pi but did not change SP. The ATP was unchanged initially and slowly decreased to the lower level during sustained secretion. These findings suggest that a sustained secretion requires more energy from ATP hydrolysis rather than initial secretion.

Acetylcholine↗
PubMed1987

Active and inactive renin-like enzymes in the brain of spontaneously hypertensive rat.

Renin-like enzyme(s) in the brain of spontaneously hypertensive rat (SHR) were activated unequivocally by trypsin. The highest concentration of the active renin-like enzyme was localized in the hypothalamus (1.03 +/- 0.25 ng angiotensin I/mg of protein per h, mean +/- S.D.), followed by the striatum (0.51 +/- 0.21), thalamus (0.40 +/- 0.08), midbrain (0.33 +/- 0.04), medulla oblongata (0.25 +/- 0.01), cerebral cortex (0.21 +/- 0.03), and cerebellum (0.14 +/- 0.03), while the highest concentration of the inactive renin-like enzyme was localized in the hypothalamus (0.86 +/- 0.17), followed by the striatum (0.47 +/- 0.15), thalamus (0.32 +/- 0.09), cerebellum (0.29 +/- 0.04), midbrain (0.26 +/- 0.02), cerebral cortex (0.24 +/- 0.04), and medulla oblongata (0.10 +/- 0.03). The active renin-like activity in the thalamus of SHR was significantly lower than that of age- and sex-matched normotensive Wistar-Kyoto (WKY) rats. Furthermore, the inactive renin-like activity in the striatum, thalamus, cerebellum, midbrain, and medulla oblongata of SHR was significantly lower than that in the corresponding areas of WKY rats. Although the precise mechanisms underlying the conversion of inactive to active renin-like enzyme in the brain remain to be resolved, these results may offer a new aspect for the role of the brain renin-angiotensin system in the initiation and/or development of hypertension of SHR.

Animals↗