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Biomedical subjects

E Furuya

Publications and source records attributed to E Furuya.

At least 37 records · Page 2Linked to original sources

A new method of quantitating serum and urinary levels of 1,5-anhydroglucitol in insulin-dependent diabetes mellitus.

A new method was developed for quantitating the serum and urinary levels of 1,5-anhydroglucitol (AG), a sensitive and informative marker of glycemic control. This method utilized a combination of ODS and pyranose oxidase-immobilized columns for HPLC, and monitored hydrogen peroxide production with an electrochemical detector. We applied this method to determine the serum and urinary AG levels in 15 patients with insulin-dependent diabetes mellitus (IDDM) as well as in control subjects. Baseline separation of AG from other sugars such as glucose and myoinositol was achieved. Quantitation of AG was achieved over the range from 0.2 ng to 0.3 micrograms based upon peak heights. The serum and urinary AG levels in the IDDM patients were 4.4 +/- 8.3 mg/l and 5.1 +/- 4.3 mg/day, respectively. We found that the urinary AG to serum AG ratio showed a linear correlation with the urinary glucose level in the IDDM patients (urinary glucose (y) vs. urinary AG to serum AG ratio (x): y = 9.071x-0.991; r = 0.968, P < 0.001). This method proved efficient and reliable for quantitating urinary AG. Since determination of both the AG and glucose levels in urine gives equivalent clinical information to the serum AG level, urinary monitoring could provide a valuable addition to the available methods for assessing the glycemic status of IDDM patients.

Adolescent↗

The effect of fructose on fructose 2,6-bisphosphate level and fructose 6-phosphate, 2-kinase activity in the perfused rat liver.

In order to investigate the effect of fructose on the metabolic rates of liver glycolysis, changes in fructose 2,6-bisphosphate content and fructose 6-phosphate, 2-kinase (F6P2kinase) activity were examined using perfused rat liver. When the starved liver was perfused with 0.5 mM fructose, the fructose 2,6-bisphosphate level was 6.66 nmol/g liver, or the maximum. However, with further increases in the fructose concentration, increments in the fructose 2,6-bisphosphate level gradually decreased. F6P2kinase was inactive in the starved rat liver. By perfusion with the medium containing fructose or even without fructose, however, the enzyme was activated. The suppression of the increase in the fructose 2,6-bisphosphate level by perfusion at higher fructose concentrations was at least partly due to the inhibition of F6P2kinase by F1P.

Adenosine Triphosphate↗

[Considerations in clinical testing--implications of urinary 17-ketosteroid sulfate and 17-hydroxycorticosteroid measurements for stress research].

With the recent increase in number of patients with stress, the search is continuing for health-indicators that can be used to determine stress reliably and quickly. These indicators rely on subjective evaluations, and the establishment of an objective method of determining stress is necessary. We have developed a method involving the determination of the 17-Ketosteroid sulfates (17-KS-S) which has proved valuable for stress research. By simultaneous measurement of both 17-KS-S and 17-hydroxycorticosteroids (17-OHCS), which have different biological significance, and which originate from the adrenal gland, an organic Yin-Yang evaluation of the adrenocortical function can be made. The two values (mainly the 17-KS-S values) are closely correlated to the biological stress state as evaluated from psychosocial factors. This method should prove to be valuable in the clinic as an objective method of determining stress.

17-Hydroxycorticosteroids↗

[Primary care considerations--17-ketosteroid sulfate-17-hydroxycorticosteroid method as a clinical stress indicator].

In primary care, the management of stress-related diseases occupies a wide spectrum of prevention and treatment, from holistic health care to final cure, centered on medical health and an organic integration of body, mind, and social parameters. Inquiries are being made at examinations and interviews to obtain information on the health condition of the patient, but, they are inadequate. There is a need for an objective indicator to help in the diagnosis. With the introduction of the Stress Barometer (measurements of 17-ketosteroid sulfates and 17-hydroxy-corticosteroids) it has become possible to obtain critical information on a patient in a very short time. This method allows early detection of stress, depression state and tiredness from work which is not presently possible from blood tests. It then becomes possible to call the patient's attention to these matters and to work at increasing awareness, energy levels, and responses to the conditions that are determined in this manner.

17-Hydroxycorticosteroids↗

Leakage of cytoplasmic enzymes from rat heart by the stress of cardiac beating after increase in cell membrane fragility by anoxia.

The effects of spontaneous beating after anoxia and the pumping stress induced by a left ventricular balloon on the leakage of myocardial enzymes from the isolated perfused rat heart were investigated. Beating of the heart was arrested by perfusion with high-K+ medium. When the beating was arrested during reoxygenation after anoxia, the leakage of lactate dehydrogenase (LDH) was significantly lower than during reoxygenation with spontaneous cardiac beating. After changing from K+ arrest to spontaneous beating by perfusion with low-K+ medium during reoxygenation, the leakage of LDH increased markedly. Imposition of left ventricular wall stress on the K(+)-arrested heart by repetitive passive distension during aerobic perfusion and after 20 min and 60 min of anoxia caused LDH leakages of 1.0, 4.6 and 21.0 units/g in 30 min, respectively. Under this mechanical stress, the release of LDH as a percentage of its total myocardial activity coincided well with that of cytoplasmic aspartate aminotransferase (AST), while the percentage release of mitochondrial AST was much less. These results appeared to indicate that the leakage of cytoplasmic enzymes during reoxygenation is accelerated by cardiac beating because of fragility of the cell membranes developing during the preceding anoxia.

Adenosine Triphosphate↗

Effect of fructose on glycogen synthesis in the perfused rat liver.

The effect of fructose on glycogen synthesis was examined in the perfused liver of starved rats. With increasing fructose concentration in the perfusate, glycogen synthesis and the % a form of glycogen synthase increased to a maximum at 2 mM and then decreased, progressively. The glucose 6-P level increased with the increase in fructose concentration. On the other hand, the ATP content was unchanged at a concentration of 2 mM or less and decreased at 3 mM or more. We also showed that the stimulation of glycogen synthesis by fructose at a concentration of 2 mM or less was due to activation of glycogen synthase by accumulated glucose 6-P and that ATP depletion at a concentration of 3 mM or more caused an increase in phosphorylase a and a decrease in glycogen synthase activity even in the presence of a high concentration of glucose 6-P.

Adenosine Triphosphate↗

Ca2+-induced accumulation of pyrophosphate in mitochondria during acetate metabolism.

The mechanism of pyrophosphate (PPi) accumulation in rat liver during acetate metabolism was investigated. Perfusion of the liver with acetate in the presence of noradrenaline and glucagon induced marked accumulation of PPi (2 mumol/g of liver, 200 times that of control). In contrast, perfusion with glutamine, which generates PPi only in the cytosol, caused little accumulation of PPi, even in the presence of the two hormones. The site of PPi accumulation was shown to be the mitochondria by the finding that isolated mitochondria from the liver perfused with acetate and the hormones contained 50 nmol of PPi/mg of protein. The addition of an uncoupler to mitochondria with accumulated PPi caused gradual decrease in their PPi content, with concomitant release of a stoichiometric amount of Ca2+. Similar accumulation of PPi was observed when isolated mitochondria were incubated with acetate and Ca2+. These results show that an increase in cytosolic Ca2+ caused by the co-administration of the two hormones induced uptake of the ion into mitochondria, and that PPi accumulated in mitochondria only when it was generated in the organelles with an elevated concentration of Ca2+. High mitochondrial concentrations of Ca2+ are considered to inhibit inorganic pyrophosphatase through the formation of a stable complex, CaPPi-. Mitochondria with accumulated PPi had normal respiratory activities, and their adenine nucleotide concentrations were increased 2-fold rather than being decreased, the increases also being considered to be caused by their high concentration of Ca2+.

Acetates↗

The effect of fructose on the cellular content of adenine nucleotides in the perfused rat liver.

In order to evaluate the validity of using fructose in total parenteral nutrition, the mechanism of the resultant decrease in hepatic ATP content on fructose loading was examined using perfused rat livers. The ATP content of the livers perfused with fructose at a concentration of 2 mM or less, was about 2.8 mumol/g liver. Perfusion with 3 mM and 5 mM fructose decreased the ATP level to 1.65 and 1.50 mumol/g liver, respectively. Transient suppression of respiration occurred after the liver was perfused with fructose at a concentration of 3 mM or more, but not after 2 mM or less. Perfusion of the liver with fructose caused an accumulation of fructose 1-phosphate and a decrease in Pi. At higher concentrations of fructose, the accumulation of fructose 1-phosphate and the depletion of Pi were greater. The Pi depletion, caused by the accumulation of fructose 1-phosphate, suppressed O2 consumption and subsequently decreased the hepatic ATP content. A stoichiometric relationship was found between the total loss of adenine nucleotides in the liver and the amount of purine catabolites in the perfusate. After the administration of fructose at a concentration of 2 mM or less, however, the amount of remaining intracellular Pi was sufficient to maintain respiration and no decrease in the hepatic ATP content was observed.

Adenine Nucleotides↗

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↗

Changes in aerobic and anaerobic ATP-synthesizing activities in hypoxic mouse brain.

The changes in cerebral metabolism in mice in severe hypoxia were investigated by analyses of changes in the levels of energy metabolites and near-infrared spectrophotometric assessment of the states of hemoglobin and cytochrome oxidase. Under 4.4% O2, the contribution of anaerobic ATP production was at most about 20% of the demand. However, the cerebral ATP level was kept at the control level until about 1 min before death. Pentobarbital anesthesia, which reduced the cerebral rate of metabolism, prolonged the survival time, although anaerobic ATP production still did not support ATP demand. Under these conditions, deoxygenation of hemoglobin and reduction of cytochrome oxidase proceeded rapidly within 1 min. Hemoglobin reached the maximum state of deoxygenation in the middle phase of hypoxia, with no further change until death. However, cytochrome oxidase was reduced slowly with one phase of partial reoxidation due to increase of cerebral blood volume, and reached the completely reduced state at death. From these results it was concluded that the aerobic ATP synthesis, which supplied more than 80% of the cerebral demand, decreased gradually because of limitation of oxygen supply, and that the failure of oxidative phosphorylation to meet demand triggered the decrease in the cellular ATP level that led to death.

Adenosine Diphosphate↗

Hepatic accumulation of pyrophosphate during acetate metabolism.

Accumulation of pyrophosphate induced by acetate administration was investigated in rat liver in situ and in perfused rat liver. Intraperitoneal injection of acetate into rats increased the pyrophosphate concentration in the liver to about 2 mumol/g liver, which was 200 times that in control liver. Perfusion of liver with acetate alone did not result in accumulation of pyrophosphate. However, the further addition of a Ca2+-mobilizing hormone, such as noradrenaline or angiotensin II, together with glucagon to the perfusion medium containing 1 mM acetate caused accumulation of pyrophosphate to a similar level to that observed in vivo. Acetate, glucagon and a Ca2+-mobilizing hormone were all required for accumulation of pyrophosphate in perfused liver. Omission of Ca2+ from the perfusion medium or addition of a Ca2+-antagonist reduced the accumulation significantly. The two kinds of hormones, glucagon and an alpha-agonist, either singly or in combination, did not affect the rate of acetate utilization. These results show that liver cells accumulate a large amount of pyrophosphate during acetate metabolism at high intracellular levels of Ca2+ that can be realized by the synergistic actions of the two kinds of hormones.

Acetates↗

Function of reticuloendothelial system on ethionine induced liver injury in mice.

Phagocytic activity as a function of the reticuloendothelial system (RES) has been studied in ethionine induced liver injury by using the carbon clearance test. Liver damage in male and female mice was induced by DL- and L-ethionine injections (1000 mg/kg/day, i.p.). In both female and male mice, a single dose or three injections of DL- or L-ethionine caused increases in liver/body weight ratio, A/G ratio, GOT and GPT levels, and BSP retention. There was the decrease of the total protein levels in the serum. The degree of liver injury was more severe after three injections of DL- and L-ethionine than after a single injection of them. After a single injection of ethionine, the L-isomer induced a slightly greater response than the racemic mixture, except for BSP retention. On the other hand, phagocytic activities by the carbon clearance test were increased after a single injection or three injections of DL- and L-ethionine. That is, the K value was increased in all ethionine treated mice except for females with three injections of DL-ethionine. The alpha value was increased after three injections in DL- and L-ethionine treated males and DL-ethionine treated females. In addition, the increase in carbon uptake by Kupffer cells can be seen by light microscopy after a single injection or three injections of DL- or L-ethionine. These findings indicate that ethionine injections induce the enhancement of RES phagocytosis, although the biochemical parameters indicating liver injury are changed severely. These results support the data indicating no correlation between the alteration of RES activity and the degree of liver injury.

Alanine Transaminase↗

Anomer specificity of glucose-6-phosphatase and glucokinase.

The anomeric form of glucose produced by glucose-6-phosphatase was studied using an apparatus that specifically measures beta-D-glucose. The time course of beta-D-glucose formation from glucose-6-P by glucose-6-phosphatase is essentially linear. In the presence of mutarotase, this rate is reduced to 70% of that obtained in the absence of mutarotase. When detergent treated microsomes were used, the rate of beta-D-glucose formation is unaffected by mutarotase. These results suggest that only beta-anomer of glucose is produced by microsomal glucose-6-phosphatase and this specificity is determined by translocase for glucose-6-P or glucose. It was also demonstrated that alpha-D-glucose is the substrate for glucokinase.

Animals↗

Studies on alterations in blood coagulative and fibrinolytic activities after single and multiple administrations of carbon tetrachloride in mice.

The time course of alterations in coagulative and fiblinolytic activities was studied in CCl4-induced liver disease in mice. Liver disease was induced by administration of 20% CCl4 in olive oil (p.o.). After single administration of CCl4, significant prolongation of r and k values and decrease in the ma value of thromboelastogram and apparent prolongation of PT and PTT were seen at 24 hr. Fibrinogen content decreased from 12 to 72 hr after single administration, while a mild but significant decrease in fibrinogen was observed after multiple administrations. The activity of factor XIII increased from 5 to 12 hr and then decreased from 24 to 168 hr after single administration. The activity of the hepaplastintest and Antithrombin III decreased apparently after single and multiple administrations. The plasminogen content and the activity of alpha 2-plasmin inhibitor decreased severely after single and multiple administrations. These results indicate that the coagulative and fibrinolytic activities were decreased to the most lowest value at 24 or 48 hr after single administration of CCl4, and the severe suppression of fibrinolysis and the mild decrease in coagulative activity were observed after multiple administrations of CCl4. The reason for the different effects between single and multiple administrations on coagulative and fibrinolytic systems was discussed.

Acute Disease↗

Management of inflammation: the parametrical system, its practical and methodological position.

Since individual cases of inflammation differ one from another, and it is necessary to distinguish between adequate and inadequate inflammatory responses, the proper Management of Inflammation demands the simultaneous assessment of the various component processes. The parameters chosen for such diagnosis should be of broad specificity and show unequivocal and consistent changes wherever the inflammatory response is inadequate. Our long-term studies of carrageenin-induced inflammation in variously conditioned rats have led to a singling out of a set of parameters which satisfy the above three conditions, some fully and some partly: lipid peroxide, proamidase and lysosomal enzymes in exudate, sialoglycoproteins in serum and alkaline phosphatase, elastase in granuloma pouch tissue. Use of this system affords a broad reliable observation of inflammatory processes which an interpretation of isolated components alone could not provide, and determines the position and significance of other components in the inflammatory system. Moreover, it permits the discovery of correlations which would hardly be realized if single mechanisms were considered, and helps to form new hypotheses and to outline fields for further study.

Alkaline Phosphatase↗

An assay method for cyclic AMP using high-performance liquid chromatography with pretreatment by alkaline phosphatase.

A convenient method for determination of cyclic AMP is described. This nucleotide was determined using high-performance liquid chromatography after the treatment of tissue extracts by alkaline phosphatase. Tissue extracts contain large amounts of materials which interfere with the measurement of cyclic AMP on the chromatogram. By the alkaline phosphatase treatment, these materials were completely converted to compounds which no longer interfered with the measurement. This method enables the detection of 2 pmol cyclic AMP, and is applicable to various tissue extracts which contain at least 0.1 nmol cyclic AMP/g wet wt.

3',5'-Cyclic-AMP Phosphodiesterases↗

Function of reticuloendothelial system on CCl4 induced liver injury in mice.

Phagocytic activity as a function of the reticuloendothelial system (RES) has been studied in CCl4-induced liver injury by using the carbon clearance test. Liver damage in mice was induced by administration of 20% CCl4 in olive oil (p.o.). After a single administration of CCl4, significant increases in liver/body weight ratio, serum GOT and GPT levels, alpha, beta and gamma-globulins and BSP retention, and decreases in serum albumin, an activity of the hepaplastintest and the correct phagocytic activity, alpha value, were found. After 15 administrations of CCl4 (3 times a week), mild increases in serum GPT level and BSP retention and decreases in the activity of the hepaplastintest and both phagocytic indices, K and alpha values, were observed. However, zymosan treatment 3 days before sacrifice induced an increase in K value depressed by multiple administrations of CCl4. The depression of carbon uptake by Kupffer cells can be seen by light microscopy after multiple administrations of CCl4 compared with that of saline and olive oil. These findings indicate that the RES phagocytosis is suppressed more strongly in chronic liver injury by 15 CCl4 administrations than in acute injury by a single one, although the biochemical parameters indicating liver injury are shown to have an opposite tendency. A clear correlation between the alteration of RES activity and the degree of liver injury was not noted.

Alanine Transaminase↗