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Biomedical subjects

K Fukui

Publications and source records attributed to K Fukui.

At least 343 records · Page 19Linked to original sources

Calcitonin gene-related peptide(CGRP) stimulates the release of atrial natriuretic peptide(ANP) from isolated rat atria.

The effect of calcitonin gene-related peptide(CGRP) on the release of atrial natriuretic peptide(ANP) was studied in spontaneously beating, isolated rat atria. CGRP stimulated the ANP release in a dose-dependent manner. When the atria were incubated with a combination of phentolamine, propranolol, and atropine, these antagonists blocked neither the rise in ANP release nor the positive chronotropic and inotropic effects of CGRP. Therefore, we conclude that CGRP stimulates ANP release as well as cardiac contractility independently of adrenergic and cholinergic receptors.

Animals↗

Molecular cloning and sequence analysis of cDNA encoding human kidney D-amino acid oxidase.

cDNA clones encoding D-amino acid oxidase were isolated from a human kidney cDNA library by hybridization with cDNA for the pig enzyme. The cDNA insert of 2.0 kilobase pairs long provided coding information for a protein consisting of 347 amino acids. The molecular mass of the enzyme was calculated to be 39,410 Da. The amino acid sequence similarity between the pig and human enzymes is 84.4%, and among the active site residues proposed from chemical modification studies, methionine-110 of the pig enzyme was replaced by threonine. Northern blot analysis confirmed the expression of an mRNA of 2.0 kilobases encoding the D-amino acid oxidase in human kidney.

Amino Acid Sequence↗

In vivo and in vitro expression of porcine D-amino acid oxidase: in vitro system for the synthesis of a functional enzyme.

In vivo expression of D-amino acid oxidase (EC 1.4.3.3, DAO), one of the principal and characteristic enzymes of the peroxisomes of porcine kidney, was examined by use of cloned complementary DNA [Fukui, K., Watanabe, F., Shibata, T., & Miyake, Y. (1987) Biochemistry 26, 3612-3618]. RNA blot hybridization analysis revealed that DAO is expressed abundantly in kidney and liver, is expressed significantly in brain, but is not expressed in lung of pig. Three mRNA species were expressed in kidney and liver, but only one was detected in brain. These results show the presence of tissue-specific regulation of DAO gene expression. In vitro expression of a functional enzyme was achieved through the construction of a recombinant plasmid containing an SP6 promoter and a restriction enzyme fragment of cDNA to generate a DAO-specific RNA transcript. The in vitro translation product of the capped RNA transcript showed significant catalytic activity, which was inhibited strongly by benzoate, a potent inhibitor of DAO. The kinetic properties of the in vitro synthesized enzyme were comparable to those of the purified enzyme from porcine kidney. It is now possible to synthesize a functional D-amino acid oxidase in vitro and to investigate its structure-function relationships.

Animals↗

Secreted proteins of human monocytes. Analysis by two-dimensional gel electrophoresis and effect of lipopolysaccharide.

A monocyte-rich preparation from the adherent cell fraction of human peripheral blood leukocytes was incubated for 1-8 h with [35S]methionine or [3H]leucine in the presence and absence of bacterial lipopolysaccharide (LPS). The macromolecules released into the supernatant were analysed by two-dimensional gel electrophoresis and radioautography. A complex labelling pattern involving at least 20 easily demonstrable and apparently distinct products with a broad range of molecular masses and isoelectric points was observed. LPS or LPS plus actinomycin in combination markedly stimulated the labelling and release of at least twelve different macromolecules ranging in apparent Mr from 12,000 to 46,000. Studies with monocytes that had been additionally purified by centrifugal elutriation and with the monocyte-like human cell line U-937 indicated that monocytes rather than contaminating cells were the source of these products. The majority of the secreted products were unique and did not cross-react with antibodies to interleukin 1 or tumour necrosis factor. The high resolving capacity of two-dimensional gel electrophoresis may be useful to define further the diverse biological activities and potential monokines released from monocytes at various stages of their differentiation and activation.

Blood Proteins↗

Positive imaging of venous thrombi and thromboemboli with Ga-67 DFO-DAS-fibrinogen.

A newly developed thrombus imaging agent, 67Ga-DFO-DAS-fibrinogen (67Ga-fibrinogen), was used for 22 studies in 20 cases of suspected deep venous thrombosis. Increased accumulation of 67Ga-fibrinogen in venous thrombi was depicted at 48 h after injection in 10 of the 15 cases (10 of 17 studies) who showed abnormal findings in radionuclide venography. A hot spot in the lung emboli was visualized in two cases. Seven of the eight cases having anticoagulant therapy showed increased 67Ga-fibrinogen uptake, while follow-up 67Ga-fibrinogen scintigraphy after the administration of heparin and urokinase did not reveal an abnormal hot spot in one case. 67Ga-fibrinogen can be made available simply by adding 67Ga solution to a vial containing fibrinogen-DAS-DFO conjugate. In conclusion, 67Ga-fibrinogen is considered to be a promising agent for detecting active venous thrombi and to assess the effect of anticoagulant therapy.

Adult↗

Endothelium-dependent contraction induced by nicotine in isolated canine basilar artery--possible involvement of a thromboxane A2 (TXA2) like substance.

The present experiments were undertaken to determine whether the response to nicotine in the isolated canine cerebral artery is endothelium-dependent. Changes in the tension of arterial strips were recorded isometrically. Removal of the endothelium was carried out by gentle rubbing, and confirmed by scanning electron microscopy. Rubbing procedure did not affect the contractile response of the strips to serotonin. Treatment of unrubbed strips with nicotine (10(-4)M) caused a transient contraction. This response was abolished by removal of endothelium and attenuated by hexamethonium (5 x 10(-6)M) and atropine (10(-6)M). The nicotine-induced contraction was attenuated also by aspirin (5 x 10(-5)M), a cyclooxygenase inhibitor, OKY-046 (5 x 10(-5)M), a thromboxane A2 (TXA2) synthetase inhibitor and ONO-3708 (5 x 10(-9)M), a TXA2 antagonist. These results indicate that the nicotine-induced contraction in canine cerebral artery is endothelium-dependent, and suggest that the endothelium-derived contracting factor (EDCF) in the nicotine-induced response is a TXA2-like substance.

Animals↗

Determination of cytopathogenic effects of Trichomonas vaginalis on cultured cells by assaying 51Cr release.

By use of an assay for 51Cr release, it was attempted to determine the cytopathogenic effects of Trichomonas vaginalis on mammalian cultured cells. 51Cr was released from Na2(51)CrO4-labeled HeLa, L-929, FL, Vero and LLC-MK 2 cells. HeLa cells were most sensitive to the damage, followed by L-929, FL, Vero and LLC-MK 2 cells. The effect depended upon the number of parasite and the incubation period. The maximum 51Cr release from the labeled cells was observed at 37 degrees C and pH 6.0. These results attest that this assay is reliable for determining the cytopathogenic effects of T. vaginalis on the target cells.

Animals↗

Effect of changes in sodium balance on renin, angiotensinogen and atrial natriuretic factor messenger RNA levels in rats.

The expression of messenger (m)RNA for renin, angiotensinogen and atrial natriuretic factor (ANF) was investigated in rats on different sodium intakes. Messenger RNA was measured by a radiodensitometric hybridization assay. In the high-sodium state, renal renin mRNA decreased, but it increased in the low-sodium state. A further increase in renin mRNA was seen in the low-sodium state after captopril administration. Angiotensinogen mRNA levels in the liver, kidney and brain were altered by varying sodium intake. In the high-sodium state angiotensinogen mRNA decreased, but in the low-sodium state it increased. After treatment with captopril, angiotensinogen mRNA levels decreased in the liver and kidney. Angiotensinogen mRNA showed tissue specificity for expression, especially in the brain. Atrial ANF mRNA levels changed slightly with different levels of sodium intake.

Angiotensinogen↗

Effects of calcium and ouabain on the release of atrial natriuretic factor.

In order to examine the role of calcium in the secretory process of atrial natriuretic factor (ANF), we studied the effects of hypercalcaemia and ouabain on the plasma concentration of immunoreactive ANF, and the effect of calcium on immunoreactive ANF release from isolated rat atria. Anaesthetized dogs were treated with CaCl2 infusion, ouabain or phenylephrine injection. With CaCl2 infusion, serum calcium and plasma immunoreactive ANF respectively increased to three and four times their basal levels. Ouabain increased plasma immunoreactive ANF to two and a half times the initial level. Neither CaCl2 nor ouabain produced any effect on right atrial pressure and heart rate, but they both significantly increased arterial pressure. Phenylephrine caused a greater increase in arterial pressure than both CaCl2 and ouabain. However, there was no significant increase in plasma immunoreactive ANF. Moreover, calcium stimulated the release of immunoreactive ANF from isolated rat atria. These results suggest that the calcium may play a key role in the secretory process of ANF.

Animals↗

Kinetic study of a change in intracellular ATP level associated with aerobic catabolism of ethanol by Streptococcus mutans.

Streptococcus mutans, a group of lactic acid bacteria and a normal inhabitant of the human oral cavity, generates ATP by substrate-level phosphorylation coupled to oxidation of ethanol (an end product of fermentation of sugars) into acetate in the presence of oxygen (K. Fukui, K. Kato, Kodama, H. Ohta, T. Shima moto, and T. Shimono, Proc. Jpn. Acad. 64B:13-16, 1988). Kinetic measurements were made of the cellular responses of S. mutans FA-1 to ethanol in comparison with those to glucose. In contrast to oxygen-independent acid production from glucose, oxygen was absolutely required for acid production from ethanol. Ethanol elicited a marked increase in the intracellular ATP concentration (ATPi) from a starved level to a steady level which was held constant as long as oxygen was present in the medium. Once oxygen was exhausted, ATPi returned to the starved level without delay. On the contrary, ATPi changes induced by glucose, which were independent of oxygen, followed a rather complicated time course before a steady level was established. Both the steady ATPi and the rate of accompanying oxygen consumption were functions of the ethanol concentration. These two parameters were linearly correlated, indicating that the unimolecular ATP turnover rate, which is independent of the rate of ATP generation in the steady state, can be calculated for cells energized by ethanol. The estimated turnover rate was 1.5 s-1 at 37 degrees C, which is comparable to that for other bacteria energized by glucose under nongrowing conditions.

Adenosine Triphosphate↗

Sodium balance effects on renin, angiotensinogen, and atrial natriuretic polypeptide mRNA levels.

The expressions of the renin, angiotensinogen, and atrial natriuretic polypeptide (ANP) genes were studied in rats with varying sodium intakes. mRNAs for angiotensinogen and renin were measured by a sensitive radiodensitometric hybridization assay. ANP mRNA was determined by a dot-blot hybridization assay. In the high-sodium state, plasma renin concentration (PRC), renal renin concentration (RRC), and renal renin mRNA decreased by 88, 90, and 75%, respectively. In the low-sodium state, PRC, RRC, and renin mRNA increased 17-fold, 2.5-fold, and 4.5-fold, respectively. With captopril treatment during sodium depletion, PRC and renin mRNA increased 144-fold and 17.1-fold, respectively, and RRC increased 4.2-fold. The angiotensinogen mRNA level in the liver and kidney decreased in the high-sodium and increased in the low-sodium state to similar extents. Captopril treatment decreased angiotensinogen mRNA in both organs by about half. Plasma ANP and ANP mRNA decreased in the low- compared with the high-sodium state, whereas atrial ANP content was unaltered in both low- and high-sodium states. These results demonstrate that sodium intake affects the expression of the renin and angiotensinogen genes and slightly alters the expression of ANP gene.

Aldosterone↗

Effects of hypercalcemia and ouabain on plasma atrial natriuretic polypeptide in anesthetized dogs.

For the purpose of examining the role of calcium ion in the secretory process of atrial natriuretic polypeptide (ANP), we studied the effects of hypercalcemia and ouabain on plasma concentration of immunoreactive ANP (irANP). Pentobarbital sodium-anesthetized dogs were treated with calcium chloride infusion (0.136 mmol.kg-1.min-1, 10 min) or ouabain injection (30 micrograms/kg), and plasma irANP concentration, right atrial pressure, mean arterial pressure, heart rate, and serum calcium concentration were measured. To evaluate the effect of rising arterial pressure, plasma irANP concentration was also measured in dogs treated with phenylephrine hydrochloride (10 micrograms/kg). With calcium chloride infusion, serum calcium concentration and plasma irANP concentration, respectively, increased to about three times and about four times their basal levels. This increase of plasma irANP concentration was not attenuated by pretreatment with adrenoceptor blockers. Ouabain increased plasma irANP concentration to approximately 2.5 times the initial level. Neither calcium chloride nor ouabain produced any effect on right atrial pressure and heart rate, but both significantly increased mean atrial pressure. Phenylephrine caused a greater increase in mean arterial pressure than both calcium chloride and ouabain. However, there was no significant increase in plasma irANP concentration. These results suggest that the calcium ion may play a key role in the secretory process of ANP.

Anesthesia, General↗

Hormonal and renal effects of atrial natriuretic peptide in patients with secondary hypertension.

To investigate the involvement of atrial natriuretic peptide (ANP) in secondary hypertension, we examined hormonal and renal responses to ANP infusion (0.025 microgram/kg/min) in 27 patients with renal parenchymal hypertension, 10 with primary aldosteronism, 8 with renovascular hypertension, and 15 normotensive subjects. The preinfusion plasma concentration of ANP was significantly higher in patients with renal parenchymal hypertension (120 pg/ml, p less than 0.01) and in patients with primary aldosteronism (98 pg/ml, p less than 0.05) than in the normotensive subjects (40 pg/ml), but it was not greater than in the patients with renovascular hypertension (73 pg/ml, NS). In the patients with renal parenchymal hypertension, plasma ANP correlated negatively with creatinine clearance (r = -0.76, p less than 0.001). Mean blood pressure (-5%, p less than 0.01) and plasma aldosterone (-40%, p less than 0.001) decreased to a similar degree in the four groups during ANP infusion. However, an increase in urinary sodium excretion caused by ANP was higher in the hypertensive than in the normotensive patients (+250% vs. +70%, p less than 0.01) and correlated positively with mean blood pressure during ANP infusion (r = 0.47, p less than 0.001). The removal of adenomas in the patients with primary aldosteronism significantly lowered both plasma levels of ANP and cyclic guanosine 2',3'-monophosphate and reduced an increase in sodium excretion during ANP infusion, whereas the responses of blood pressure and plasma aldosterone to ANP infusion were not altered by the operation. Thus, these results suggest that elevated ANP secretion and increased natriuretic responses to ANP may modify the blood pressure and body fluid volume status in some types of secondary hypertension.

Adrenal Gland Neoplasms↗

Effects of an antihypertensive vasodilator, pinacidil, on regional blood flow in conscious spontaneously hypertensive rats.

Using a radioactive microsphere method, the effects of hydralazine and a new antihypertensive vasodilator, pinacidil, on systemic and regional hemodynamics were examined and compared in conscious Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). A single intravenous administration of pinacidil decreased mean blood pressure in a dose dependent fashion (0.03-0.3 mg/kg), and the peak effect was observed within a few minutes after administration. The onset of the hypotensive action of hydralazine was relatively slow compared to that of pinacidil, and its peak effect appeared 5-10 min after administration. There was no difference between WKY and SHR in the time course of blood pressure change, but the pressure reduction in WKY was less than that in SHR. We observed regional hemodynamic differences between the normotensive rats and the hypertensive rats. The flow rates in the kidney, spleen, gastrointestinal tract and skin of SHR were significantly lower than those of WKY. The intravenous infusion of pinacidil at a rate of 0.06 mg/kg/min increased cardiac output and heart rate, and selectively increased flow rates in organs with low flow rates compared to normotensive controls except for the spleen. Thus, pinacidil reduced total peripheral resistance of SHR to levels of WKY and normalized hemodynamic abnormalities in SHR. The regional hemodynamic action of pinacidil in SHR was not qualitatively different from that of hydralazine except in a few organs.

Animals↗

Renal effects of leukotrienes C4 and D4 in anesthetized dogs.

We investigated the effects of leukotrienes (LTs) C4 and D4 on renal function and hemodynamics of dogs. LTC4 (0.5 microgram/min, infused into the renal artery), but not LTD4 (1 microgram/min), caused an increase in renal blood flow, urine flow, urinary excretion of sodium and potassium, and rate of free water reabsorption, with a concomitant rise in systemic blood pressure. Intrarenal infusion of LTC4 under conditions of constant renal perfusion pressure induced no significant effect on renal blood flow or urine formation. Infusions or LTC4 and LTD4 into the renal artery induced no change in the release of prostaglandins, PGE2, PGF2 alpha, 6-keto-PGF1 alpha or thromboxane B2 into renal venous blood. From these findings, we concluded that the renal effects of LTC4 were secondary responses to a rise in systemic blood pressure.

Anesthesia↗

Optimal protein intake estimated by the resistance to streptococcal infection and the nutritional indices in mice.

Effects of dietary protein levels on the resistance against a bacterial infection and on the nutritional status were studied in mice to obtain basic data for the estimation of an optimal protein intake. Female DDY strain weanling mice were fed 5, 7, 10, 20, 30, or 40% casein diet. At 2 or 4 weeks on the diets, the mice were injected intraperitoneally with 5 x 10(3) or 5 x 10(4) group B streptococci/g body weight and their survival rates were observed for the following 10 days. Nutritional indices and cell numbers of thymus and spleen were also measured. The survival rate was higher in the order of 7, 10, 20, 30, 5, 40% casein diet group. Significant differences were observed between the 7% group and the 30, 5, or 40% group, and between the 10% group and the 5 or 40% group. The nutritional indices and cell numbers of the thymus and spleen were similar among the 20, 30, and 40% casein diet groups and decreased in the order of 10, 7, 5% casein diet groups. From the results, protein levels were categorized into 4 groups: severe protein deficiency with low resistance (5% casein diet), moderate protein deficiency with high resistance (7 and 10% casein diets), normal protein intake with normal resistance (20% casein diet), and high protein intake with low resistance (30 and 40% casein diets). This grouping suggests that when the relation between immunocompetence and nutritional status is considered, both high and low protein intakes are undesirable and the optimal level of dietary protein will be limited to a narrow range.

Animals↗

Effects of a synthetic atrial natriuretic polypeptide on intrarenal hemodynamics in dogs.

The effects of a synthetic human atrial natriuretic polypeptide (alpha-hANP) on the intrarenal distribution of blood flow were examined in anesthetized dogs. Intrarenal infusion of alpha-hANP at a rate of 0.05 microgram/kg per min resulted in a significant increase in renal blood flow, urine flow and urinary excretion of sodium with no change in renal perfusion pressure. Measurement of the intrarenal blood flow by the microsphere method indicated a greater increase in flow rate in the juxtamedullary than in the superficial area. A significant correlation was observed during alpha-hANP infusion between changes in both urine flow and sodium excretion and inner cortical blood flow; changes in inner cortical blood flow may reflect changes in medullary blood flow. However, a smaller dose of alpha-hANP (0.01 microgram/kg per min) increased urine flow and electrolyte excretion but had no effect on the distribution of renal blood flow. Thus, neither the increased inner cortical blood flow nor the redistribution of blood flow is the sole cause of the natriuresis during infusion of alpha-hANP. Changes in intrarenal hemodynamics could contribute to the natriuresis induced by alpha-hANP, via washout of medullary solutes.

Animals↗