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H Kohno

Publications and source records attributed to H Kohno.

At least 415 records · Page 23Linked to original sources

[Magnetic resonance imaging of experimental cerebral ischemia: correlations between NMR parameters and water content].

Recent studies on proton NMR imaging revealed its remarkable sensitivity for detecting cerebral ischemia. Since proton NMR reflects the distribution and state of water in the brain, an NMR imager becomes a sensitive in vivo detector of brain edema developing soon after the energy state is compromized by ischemia. To further clarify the usefulness of NMR imaging to characterize the ischemia-induced changes, correlations between T1 and T2 relaxation times and water content of the normal and ischemic rat and gerbil brain were studied by means of both spectroscopic and in vivo imaging methods. In the spectroscopic experiment on excised rat brain (cortex, white matter, hippocampus and thalamus for normal and ischemia-laden brain), T1 and T2 relaxation times and water content were determined. The ischemic insult was induced for 60 min by the method of Pulsinelli followed by 60 min of reperfusion. All of the T1, T2 and water content significantly increased in the ischemic tissue. Gray-white difference was evident in T1 and T1 was linearly correlated with the water content of the tissue. T2 was by far prolonged in the ischemic tissue compared with the increase in the water content, showing greater sensitivity of T2 for detection of ischemia. In the imaging experiment, coronal NMR imaging at 0.5 tesla was performed employing proton density-weighted saturation recovery (TR = 1.6 s, TE = 14 ms), T1-weighted inversion recovery (TR = 1.6 s, TI = 300 ms, TE = 14 ms) and T2-weighted spin echo (TR = 1.6 s, TE = 106 ms) pulse sequences.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Myocardial protection in coronary occlusion by retrograde cardioplegic perfusion via the coronary sinus in dogs. Preservation of high-energy phosphates and regional function.

The lack of adequate myocardial preservation because of maldistribution of cardioplegic solution in coronary artery disease remains a perplexing problem. We compared two methods of cardioplegic delivery in dogs: antegrade aortic root perfusion (Group I) and retrograde coronary sinus perfusion (Group II). Metabolic changes and regional function in the coronary occlusion model, in which the left anterior descending artery was occluded at its prediagonal portion, were studied. In the distribution of the occluded coronary artery, adenosine triphosphate and total adenine nucleotides at the end of 120 minutes of ischemia were preserved better in Group II (16.80 and 22.94 mumol/gm dry weight) than in Group I (11.06 and 16.19 mumol/gm dry weight, p less than 0.05). Lactate accumulation tended to be higher in Group I than in Group II (114.0 and 87.2 mumol/gm dry weight, respectively; not significant). Percent recovery of segmental shortening was also better in Group II than in Group I (100% and 22.3% at the same left atrial pressure, 4 mm Hg; p less than 0.01). In the region supplied by the intact coronary artery, there were no significant differences between the two groups with regard to metabolic changes and regional function. These observations suggest that retrograde cardioplegic perfusion via the coronary sinus is preferable for surgically treatment of severe coronary artery disease.

Adenine Nucleotides↗

Inhibition by free radical scavengers and by cyclooxygenase inhibitors of the effect of acidosis on calcium transport by masseter muscle sarcoplasmic reticulum.

In vitro, arachidonic acid depressed calcium transport by sarcoplasmic reticulum (SR) in the homogenate of canine masseter muscle. This effect was inhibited by superoxide dismutase (SOD), a scavenger of the superoxide anion radial ( . O-2), at pH 7.0, and by SOD plus d-mannitol, a scavenger of hydroxyl free radical ( . OH), at pH 5.5. Indomethacin and 2-aminomethyl-4-tert-butyl-6-propionyl phenol (ONO-3144), a compound known to accelerate the conversion of prostaglandin G2 (PGG2) to PGH2 and scavenge free radicals, inhibited the effect of arachidonic acid at both pH 7.0 and pH 5.5. PGG2, but not PGH2, duplicated the effect of arachidonic acid. The effect of PGG2 on SR function was similar to that of exogenous free radicals generated from the xanthine-xanthine oxidase system. Incubation at pH 5.5, in the absence of an exogenous free-radical generating system, depressed SR calcium transport in the homogenate and in isolated SR. This effect in the homogenate was inhibited by indomethacin or by ONO-3144. At 10-min incubation at pH 5.5, SOD partially and temporarily reversed the depressant effect of acidosis. The addition of SOD plus d-mannitol completely reversed the system. d-Mannitol alone was ineffective. Arachidonic acid was able to mimic these effects of acidosis, except that arachidonic acid further depressed isolated SR calcium transport. These results demonstrate that acidosis can depress SR calcium transport in the homogenate of masseter muscle by an oxygen-free radical mechanism by the generation of . O-2 and . OH. Our results also demonstrate that significant oxygen radical generation can occur through the cyclooxygenase pathway of arachidonic acid metabolism at an acidotic pH in the cellular environment outside of the SR of the muscle cell, and seems to be responsible for the generation of the . OH derived from . O-2.

Acidosis↗

The effects of lead on differentiation of the Friend leukemia cells and rat bone marrow cells.

The effects of lead toxicity on differentiation of erythroid cells was examined using Friend leukemia cells induced with 2% dimethyl sulfoxide. By increasing the concentration of lead, these cells exhibited a lag before the onset of induction. When incorporation of [3H]-delta-aminolevulinic acid into heme was compared, the maximum incorporation into the cells without lead was on the third day after induction, while it was on the fifth day in the presence of lead (5 X 10(-4) M). However, these cells did reach similar differentiation stages by the seventh day. The amount of delta-aminolevulinic acid which was excreted into the medium increased in the presence of more than 10(-4) M lead. The amount of coproporphyrin III was slightly increased with 10(-6) M lead. Protoporphyrin IX content in the cells decreased slightly with increasing concentrations of lead. delta-Aminolevulinic acid dehydratase activity in cells which were cultured in the presence of lead was low, while synthesis of this enzyme increased in the presence of lead. The bone marrow cells from lead-poisoned rats exhibited a lag in onset of maturation, which was consistent with the effect of lead on Friend cells.

Aminolevulinic Acid↗

Properties and activities of aminopeptidases in normal and mitogen-stimulated human lymphocytes.

Human peripheral lymphocytes were found to contain at least two distinct aminopeptidases, designated cytosol aminopeptidase and microsomal aminopeptidase, which differed from one another with respect to intracellular localization, substrate specificity, metal-ion activation, Km value and electrophoretic mobility. No change in these aminopeptidase activities was observed in cultured lymphocytes in the absence of mitogen throughout the cultivation period. The addition of phytohaemagglutinin or concanavalin A to the culture medium caused, in dose-dependent manner, a significant increase in cytosol aminopeptidase activity in lymphocytes. On the other hand, no increase in microsomal aminopeptidase activity was observed under the same conditions. The biochemical properties of aminopeptidases in stimulated cultured lymphocytes were identical with those of the enzymes in peripheral lymphocytes and unstimulated cultured lymphocyte. The phytohaemagglutinin dose-response curves for lymphocyte activation as measured by the DNA synthesis rate and for cytosol aminopeptidase activity were observed to be similar. However, when DNA synthesis was temporarily blocked by hydroxyurea, the rate of increase of aminopeptidase activity was unaffected. Pokeweed mitogen only slightly increased the cytosol aminopeptidase activity in cultured lymphocytes, although the lymphocytes were highly activated.

Aminopeptidases↗

In vitro binding of 67Ga to isolated rat liver cells.

In order to understand the mechanism of the accumulation of gallium citrate (Ga-67) in normal liver, an in vitro investigation system was developed using isolated rat liver cells, and various basal factors relating to 67Ga binding to normal rat liver cells were studied. In this study, the values of 67Ga binding to the liver cells increased in parallel with the number of cells; however, binding was hardly affected by higher doses of 67Ga. The binding of 67Ga to the cells was inhibited by the presence of ethylenediaminetetraacetate (EDTA) and citrate. Phosphate and bicarbonate (10(-2) M) slightly inhibited 67Ga binding to the cells. The binding of 67Ga to the cells increased as the pH was decreased. These results suggest that 67Ga binding to normal rat liver cells may occur in 'free' gallium. Moreover, the utilization of isolated rat liver cells is useful for understanding the 67Ga accumulation mechanism in normal liver.

Animals↗

Comparison of renal responses to synthetic human PTH(1-34) administration in normal young and elderly male subjects.

A parathyroid hormone (PTH) loading test with synthetic human PTH(1-34) was performed in 7 young and 6 elderly normal males. The elderly subjects had significantly higher mean basal levels of serum PTH than the young subjects (0.262 +/- 0.035(SE) vs 0.097 +/- 0.012 ng Eq/ml, P less than 0.001). When human PTH(1-34) at a dose of 100 U was administered to these subjects, the mean increases in urinary excretions of adenosine cyclic 3',5'-monophosphate(cAMP) and inorganic phosphorus (Pi), expressed as increases in absolute amounts per unit time, were significantly lower in the elderly subjects. (3.65 +/- 1.02 vs 7.41 +/- 1.05 mumol/h, P less than 0.05 for cAMP and 14.7 +/- 6.3 vs 41.8 +/- 8.6 mg/2h, P less than 0.05 for Pi) and an inverse correlation was found between the serum PTH levels and the increases in urinary cAMP excretion (mumol/hr; r = -0.63, P less than 0.05). However, when corrected for the glomerular filtration rate (GFR) and the units of PTH administered per kg body weight, the increases were not significantly different in the two groups (elderly 52.1 +/- 7.5 vs young 60.0 +/- 19.2 nmol . kg/100 ml GFR . U . h for cAMP and 0.315 +/- 0.061 vs 0.186 +/- 0.044 mg . kg/100 ml GFR . U . 2 h respectively for Pi).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Effect of saikosaponin derivatives upon the immune response against T-dependent and T-independent antigens in mice.

Effect of water soluble extracts and purified derivatives from the root of the plant Bupletum falcatum L. upon the immune response of BALB/c mice was investigated using heterologous erythrocytes and bacterial lipopolysaccharide. This Chinese Medicine is famous for its reputed anti-inflammatory, anti-tumor and other biological activities, and has been widely used alone or in a complexed form since ancient times in Oriental Countries. The effect on immune responsiveness of this drug was judged by the measurement of antibody secreting cells causing localized hemolysis in agar gel. The purified derivatives, Saikosaponin (SS) a and d, suppressed anti-SRBC plaque-forming cells (PFC), but the other derivatives, b1 and b2 and c, did not. Moreover, derivatives a and d also enhanced anti-LPS PFC. From experiments establishing minimum effective doses for anti-SRBC PFC in mice, one mg/kg was optimum for BALB/c mice. The effect of the drug on antibody formation against heterologous erythrocytes was nonspecific because treated mice had greatly depressed PFC responses against both sheep and rabbit erythrocytes. The phenotypic effect of this drug as judged by PFC against T-dependent or T-independent antigen was not identified. However, it was suggested that the purified derivatives SSa and d stimulated both T- and B-cells. The possible mechanism of effect against immunocompotent cells is discussed.

Animals↗

Correlations between proton nuclear magnetic resonance imaging and retrospective histochemical images in experimental cerebral infarction.

Evaluation of ischemic brain injury in experimental cerebral infarction in gerbils and rats was performed by means of both proton nuclear magnetic resonance imaging ([1H]NMR-CT) and various histochemical analyses. In vivo nuclear magnetic resonance (NMR) imaging was carried out employing saturation recovery, inversion recovery, and spin echo pulse sequences. Spatial resolution of the images was excellent. The ischemic lesions were detected with a remarkable contrast in inversion recovery and spin echo images within a few hours after insult. Those changes in NMR images consistently corresponded with the various retrospective histochemical observations, especially with methods related to brain edema (K+ staining) rather than structural (enzymatic) studies. Calculated T1 and T2 relaxation times indicated the evolution of the edema state in the brain in situ. They correlated excellently with the retrospective water content measurement. As a result, detailed characterization of the edema state induced by cerebral ischemia was possible in vivo using [1H]NMR imaging.

Adenosine Triphosphatases↗

Transferrin and iron uptake by rat reticulocytes.

The uptake of transferrin labeled with 3H and 59Fe by rat reticulocytes was studied to clarify the characteristics of the uptake process and intracellular transport. Rat reticulocytes took up transferrin in a saturable, time- and temperature-dependent manner. Scatchard analysis of the binding parameters indicated that transferrin molecules were bound to cell-surface receptors with high affinity. Monodansyl- cadaverine, a potent inhibitor of transglutaminase, reduced the amount of internalized transferrin but has no effect on the total amount of cell-associated transferrin, suggesting that transferrin is taken up by rat reticulocytes via receptor-mediated endocytosis. About 50% of the internalized 3H label was released from the cells after reincubation for 1 h in fresh medium. In contrast, no release of 59Fe label was observed. By immunoprecipitation and subsequent SDS-PAGE the released 3H-labeled product was identified as apotransferrin. Lysosomotropic reagents and a proton ionophore reduced the uptake of 59Fe. These results indicated that iron was removed from transferrin at an intracellular site in an acidic environment. The released iron was found not to associate with any intermediate ligands before it was utilized for heme synthesis in mitochondria.

Acyltransferases↗

Expression and phosphorylation of transferrin receptors in mitogen-activated peripheral blood lymphocytes.

Expression of transferrin receptors of cultured human lymphocytes has been investigated by using monoclonal antibody (5E9) specific for human transferrin receptors. When isolated lymphocytes were cultured in a medium containing fetal calf serum, the biosynthesis of transferrin receptor was barely detectable. The addition of concanavalin A or human serum to the medium caused a slight stimulation of the biosynthesis. The addition of concanavalin A and human serum in combination caused the highest biosynthetic activity. Appearance of the receptor on the cell surface increased in parallel with the degree of the synthesis. Treatment of concanavalin A- and human serum-treated cells with 12-O-tetradecanoylphorbol-13-acetate (TPA) resulted in a marked stimulation of the phosphorylation of the receptor. Enhancement of phosphorylation occurred within 20 min after the addition of TPA. The density of the receptor on the cell surface slightly increased upon TPA treatment of cells, and the treatment was without effect on iron incorporation from transferrin into the cells. The density of newly synthesized receptor in TPA-treated cells was similar to that in non-treated cells. These results indicated that TPA treatment of mitogen-activated human lymphocytes stimulated the phosphorylation of transferrin receptors, but TPA had no effect on the expression of the receptors thereafter.

Blood Physiological Phenomena↗

Tumor-promoting, phorbol ester-induced phosphorylation of cell-surface transferrin receptors in human erythroleukemia cells.

When human erythroleukemia cells (K562) were exposed to phorbol-12-myristate 13-acetate (PMA), phosphorylation of transferrin receptors was enhanced 5-fold with 10(-7) M PMA and 7-fold with 10(-6) M PMA, but not with 4 alpha-phorbol (5 X 10(-7) M). Stimulation took place in serine residues in the cytoplasmic domain of the receptor. Although phosphorylation in the control cells took place in both cell-surface and intracellular receptors, phosphorylation in PMA-treated cells increased only in the cell-surface receptors, not in the intracellular receptors. The number of receptors on the cell surface increased slightly with the increase in phosphorylation at the cell surface, in the PMA-treated cells. No difference in transferrin binding was found for the control and PMA-treated cells. These results indicate that enhanced phosphorylation of the transferrin receptor takes place on the cell surface only and that it presumably is mediated by protein kinase C.

Animals↗

Characterization of the effect of pH on the excitation-contraction coupling system of canine masseter muscle.

The effect of pH on the excitation-contraction coupling system of canine masseter muscle was studied by evaluating the functional integrity of the sarcoplasmic reticulum (SR) and myofibrils. Increasing proton concentration (pH 7.0-5.8) significantly reduced oxalate supported SR calcium uptake velocity, while Ca2+-stimulated, Mg2+-dependent ATPase activity was unaffected by pH. The efficiency ratio of calcium transport, or the coupling ratio (mumoles Ca2+ transported/mumoles ATP hydrolyzed), decreased from 1.094 +/- 0.042 at pH 7.0 to 0.946 +/- 0.036 at pH 6.0 (P less than 0.05) and to 0.780 +/- 0.024 at pH 5.8 (P less than 0.01). Myofibrillar pCa (-log [free Ca2+] )-ATPase activity was unaffected between pH 7.0 and pH 6.5. At pH 6.0, increasing Ca2+ concentration inhibited myofibrillar ATPase activity, and this inhibitory phenomenon was accentuated at pH 5.8. Kinetic analysis of the myofibrillar pCa-ATPase data, utilizing double-reciprocal plots, demonstrated an increase in Km at low pH. It is concluded that acidosis significantly uncouples calcium transport from ATP hydrolysis in the SR of masseter muscle and significantly alters myofibrillar ATPase activity. It is hypothesized that these defects may explain an observed depression in skeletal muscle cell function during ischemia.

Adenosine Triphosphatases↗

Relation between 67Ga uptake and the stage of inflammation induced by turpentine oil in rats.

For the establishment of the experimental system to judge easily the effect of anti-inflammatory drug, 67Ga-citrate was used. The weight of granuloma tissues induced by inflammable agent, turpentine oil, gradually increased and reached a maximum at 6 days after the administration of turpentine oil. Gallium-67 accumulation in the inflammatory lesions showed also a maximum at 6 days after that. Both patterns were closely similar each other. These results showed that the processes and/or stages of inflammation could be indicated by the pattern of 67Ga uptake.

Animals↗