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Biomedical subjects

H Kosaka

Publications and source records attributed to H Kosaka.

At least 127 records · Page 7Linked to original sources
PubMed1994

Direct proof of nitric oxide formation from a nitrovasodilator metabolised by erythrocytes.

When 15N-isosorbide dinitrate was injected into rats, the venous blood showed a 2-line hyperfine structure in the electron spin resonance (ESR) spectra of NO hemoglobin (HbNO), proving generation of 15N-NO. When human blood was mixed with 15N-isosorbide dinitrate anaerobically, ESR spectra showed initially 14N-HbNO, which was then overwhelmed with the increased 15N-HbNO. With deoxygenation of the blood of another 4 male donors without addition of isosorbide dinitrate, two showed 14N-HbNO, but others did not, suggesting varied amount of NO-precursers, nitrosothiols, in human blood. Neither N-ethylmaleimide nor S-hexylglutathione, a glutathione-S-transferase inhibitor, inhibited the formation of UbNO in the hemolysate mixed anaerobically with isosorbide dinitrite.

Adult↗
PubMed1994

Nitrosyl hemoglobin production during reperfusion after focal cerebral ischemia in rats.

We first detected a definite nitrosyl hemoglobin (HbNO) signal in the jugular blood by electron spin resonance spectroscopy during early reperfusion after cerebral ischemia. A distinct three-line hyperfine structure, characteristic to HbNO, was demonstrated at 30 min of recirculation after 2 h of middle cerebral artery occlusion in rats. Only a weak HbNO signal was observed in animals with 2 h sustained ischemia or with sham operation. The present findings suggest that reperfusion after cerebral ischemia facilitates nitric oxide generation in the brain, which leads to the increased nitrosylation of erythrocyte hemoglobin in the cerebral circulating blood.

Animals↗
PubMed1994

Study on measurement of delta-aminolevulinic acid in plasma by high-performance liquid chromatography.

A method for the determination of delta-aminolevulinic acid in plasma of lead-exposed workers by high-performance liquid chromatography with fluorescence detection of a fluorescent delta-aminolevulinic acid derivative (2-methylidineamino-3,5-diacetyl-4,6-dimethylpropionic acid) was established. The detection limit of delta-aminolevulinic acid in plasma was 0.01 microgram/ml at a signal-to-noise ratio of 5:1. A linear correlation was obtained between the amounts of delta-aminolevulinic acid injected from 0.01 to 0.5 microgram/ml (r = 0.999). The recovery of 0.05 and 0.1 microgram/ml of delta-aminolevulinic acid added to plasma with various concentrations of delta-aminolevulinic acid in plasma ranged from 80.0 to 100.8%. This method, combined with the use of an automatic sampler, should facilitate the routine measurement of delta-aminolevulinic acid in plasma.

Aminolevulinic Acid↗
PubMed1994

Effect of interferon-gamma on nitric oxide hemoglobin production in endotoxin-treated rats and its synergism with interleukin 1 or tumor necrosis factor.

We studied the in vivo effect of interferon-gamma (IFN-gamma) on nitric oxide (NO) generation. ESR spectra of nitric oxide hemoglobin (HbNO) appeared after a lag time of 2h in the blood of rats treated with Escherichia coli lipopolysaccharide (LPS). IFN-gamma enhanced LPS-induced HbNO formation in rats without modifying the time lag, although IFN-gamma alone did not induce HbNO formation. The plasma nitrate concentration was approximately one order of magnitude higher than the HbNO concentration. On treatment with LPS alone, the amount of tumor necrosis factor (TNF) released decreased after 2 h. Simultaneous addition of IFN-gamma and LPS increased TNF release for at least 8 h. Interleukin 1 (IL-1) release was detected only at 2 h in both groups. We also investigated the in vivo interactions of these cytokines. TNF plus IL-1 induced the greatest HbNO generation, followed by TNF plus IFN-gamma, and then IL-1 plus IFN-gamma. These results suggest that increase of TNF release by IFN-gamma plays a key role in NO generation in LPS-treated rats.

Animals↗
PubMed1994

Intravital microreflectometry of individual pial vessels and capillary region of rat.

A microscopic reflectance spectrophotometer was constructed to obtain the spectra of single pial vessels and of a region containing only capillaries (capillary region). The difference in the oxygen saturation (SO2) of hemoglobin between the regional arteriole and venule [R(A - V)] and that between the regional arteriole or capillaries [R(A - C)] were calculated. The reduction of cytochrome aa3 was also estimated in the capillary region. This method was applied to the brain surface of spontaneously breathing rats subjected to hypoxic and anemic hypoxia. On decreasing the inhaled O2 from 100 to 15%, elevation of R(A - V) and R(A - C) with slight arteriolar dilatation (though statistically not significant) was observed. Below 10% O2 (especially at 4 and 3% O2), the R(A - V) and R(A - C) decreased in spite of significant arteriolar dilatation with progressive reduction of cytochrome aa3, indicating suppression of oxygen transport to mitochondria. In the case of hemodilution down to 37% hematocrit (Ht), elevation of R(A - V) and R(A - C) occurred with a slight tendency toward arteriolar dilatation. Below 32% Ht, the R(A - V) decreased but the R(A - C) remained steady, while reduction of cytochrome aa3 progressed. Altogether, the SO2 in the capillary region decreased and the reduction of cytochrome aa3 progressed with the decline of arteriolar O2 supply in both hypoxic and anemic hypoxia.

Anemia↗
PubMed1994

Elevation of plasma nitric oxide end products during focal cerebral ischemia and reperfusion in the rat.

We investigated the alterations in the stable end products of nitric oxide, i.e., nitrate and nitrite, in the plasma during and after rat focal cerebral ischemia by an automated procedure based on the Griess reaction. At 2 h of middle cerebral artery (MCA) occlusion, plasma nitrate/nitrite levels were significantly higher (53 +/- 8 microM, mean +/- SD, n = 5, p < 0.05) than in rats with sham operation (36 +/- 9 microM, n = 5), and were mildly elevated at 4 h of MCA occlusion (42 +/- 9 microM, n = 5, n.s.). At 30 min of reperfusion after 2 h of MCA occlusion, plasma nitrate/nitrite levels were more markedly elevated (72 +/- 7 microM, n = 5, p < 0.01 vs. sham operation), but were moderately elevated at 2 h of reperfusion after 2 h of MCA occlusion (61 +/- 10 microM, n = 5, p < 0.05). Plasma nitrite levels were not changed during these experimental periods. Administration of 20 mg/kg of NG-nitro-L-arginine methyl ester (L-NAME) significantly decreased plasma nitrate/nitrite as well as nitrite at 30 min of reperfusion after 2 h of MCA occlusion (n = 5), but 2 mg/kg of L-NAME did not (n = 3). The effect of 20 mg/kg of L-NAME on plasma nitric oxide end products was reversed by the simultaneous administration of 200 mg/kg of L-arginine (n = 3), but not D-arginine (n = 3). The present study suggests that the L-arginine-nitric oxide pathway is activated during acute cerebral ischemia and reperfusion.

Animals↗
PubMed1994

Microspectroscopic measurement of the optical properties of rat liver in the visible region.

Microspectroscopy is used to investigate optical properties of haemoglobin-free perfused rat liver. Visible spectra of 20 microns diameter spot size were measured in transmission and/or reflection modes as a function of the thickness (< 1200 microns) of the liver-edge. Optical density (OD) in transmission mode increased with the increasing liver thickness, whereas in reflection mode OD decreased but became almost constant above a certain thickness (c.600 microns) of the liver. The Kubelka-Munk (KM) two-flux model, with a minor modification, was applied successfully to the analysis of the changes in OD as a function of the thickness. This approach estimates the KM absorption coefficient (EKM), KM scattering coefficient (SKM) and effective penetration depth (delta eff) of the liver. The optical properties were similar to reported values, obtained with different methods.

Animals↗
PubMed1994

ESR spectral transition by arteriovenous cycle in nitric oxide hemoglobin of cytokine-treated rats.

Nitric oxide (NO) generation was induced in rats by Escherichia coli lipopolysaccharide (LPS) as detected by electron spin resonance (ESR) signals of NO hemoglobin (HbNO). However, there were inconsistencies in ESR spectral shape among them. We have therefore carried out a systematic study to clarify the in vivo spectral changes. First, the spectra of the alpha-NO heme species had the distinct three-line hyperfine structure in venous blood but not in arterial blood in all rats treated with tumor necrosis factor, interleukin-1, and/or LPS, and methemoglobin was not detected at the g = 6 (high-spin methemoglobin) region. Second, when the treated rats died, the three-line hyperfine structure was very distinct even in arterial blood. Third, even if HbNO was formed by injection of nitrite to rats, the three-line hyperfine structure of HbNO in venous blood was more marked than that in arterial blood, independent of the appearance of the methemoglobin signal. Fourth, an ex vivo study using whole blood demonstrated that the three-line hyperfine structure intensified lineally when O2 saturation of hemoglobin decreased but disappeared on reoxygenation of hemoglobin. These results directly demonstrate in vivo quaternary structural transition of the hemoglobin tetramer from the high-affinity state in the arterial cycle to the low-affinity state in the venous cycle. The transition makes the diverse ESR spectra of HbNO in vivo.

Animals↗
PubMed1994

Assessment of rate of O2 release from single hepatic sinusoids of rats.

Characteristics of O2 release from single hepatic sinusoids are described in relation to the sinusoidal O2 inflow for the first time. Microspectroscopic measurement of the rate of O2 release (RO2) from single sinusoids to surrounding hepatocytes was carried out at the edge of the liver of anesthetized rats. The oxyhemoglobin inflow ([HbO2] inflow) into sinusoids, which is a product of sinusoidal blood flow rate and inflowing oxyhemoglobin concentration ([HbO2]), was varied by exchange transfusion of blood with physiological saline. 1) RO2 from single hepatic sinusoids of control rats was fairly constant [0.66 +/- 0.14 (SD) nmol.cm-2.s-1, with 18 sinusoids of 9 rats] over a wide range of [HbO2] inflow (50-200 fmol/s). 2) In the cases of mild anemia, [HbO2] inflow scarcely decreased but RO2 increased, probably because of the increased metabolism of hepatocytes. 3) In the cases of severe anemia, [HbO2] inflow decreased below 50 fmol/s and RO2 decreased accordingly. 4) In all cases no difference was observed between the RO2 values obtained from periportal zone and those from pericentral zone. These results are discussed on the bases of the vascular structure and function of hepatic sinusoids and on the animal model of hemorrhagic shock.

Acute Disease↗
PubMed1994

Reactive oxygen species involved in phenazine-methosulfate-induced rat lens opacification. An experimental model of cataract.

The excised rat crystalline lens opacified when incubated aerobically with phenazine methosulfate, but no opacification was observed under anaerobic conditions. Morphological studies revealed development of opacification in the cortex. The opacification resembled that often seen in the early period of senile cataract as well as in naphthalene-induced and UV cataract. Both an increase in hydration and in electrolyte imbalance accompanied this opacification. Na,K-ATPase activity of the opacified lens was found to decrease. In order to investigate if activated oxygen is involved in these processes, we conducted an electron spin resonance study by means of a spin trapping technique. When the lens homogate was incubated with phenazine methosulfate, OH radicals were generated under aerobic but not under anaerobic conditions. Reduced pyridine nucleotides must be involved in the process, because the mixture of nicotinamide adenine dinucleotide phosphate [NAD(P)] and phenazine methosulfate did not generate OH radicals, but the mixture of NAD(P)H and phenazine methosulfate generates OH radicals, indicating that reduced phenazine methosulfate was involved in the OH radical generation. Probably, the generated OH radicals inactivated Na,K-ATPase residing in the epithelium of the lens, which eventually caused opacification of the lens. The present experiment system may be used for the elucidation of lens opacification (cataract) involved with reactive oxygen species.

Animals↗
PubMed1994

[The SOS-inhibition activity of human urine investigated by umu-test--diurnal and daily changes of SOS-inhibition activity of urine].

We found that human urine suppressed SOS-responses induced by furylfuramide (AF-2), as detected by the umu test using Salmonella typhimurium TA1535/pSK1002. In the present report, we studied the time stability of the SOS-inhibition activity in urine. The diurnal and daily changes of SOS-inhibition in the urine were also observed. Results obtained were as follows; 1) SOS-inhibition activity of the urine remained stable more than one month after the urine was frozen. 2) Individual variation was observed in the SOS-inhibition activity of the urine. 3) Total SOS-inhibition activity of per a day showed relatively small variation during experimental days. 4) The SOS-inhibition activity of urine was higher early in the morning than in the daytime. The activity fell gradually with time in the daytime and showed the lowest value in the evening. Then, it rose again at night. Therefore, it is necessary to collect urine at specific times to avoid the differences caused by diurnal changes in SOS-inhibition activity.

Adult↗
PubMed1993

Spin trapping of superoxide released by opsonized asbestos from human promyelocytic leukemia cell line, HL60.

By ESR using 5,5-dimethyl-1-pyrroline-1-oxide as a spin trap, superoxide (O2-) production was proved upon stimulation of dimethyl sulfoxide-differentiated HL60 by crocidolite opsonized with fresh or refrigerated serum, as well as by phorbol myristate acetate (PMA). Crocidolite, unopsonized or opsonized with frozen-thawed or heat-inactivated serum, did not induce O2- release. Addition of iron chelators or superoxide dismutase inhibited O2- release completely. Neither undifferentiated nor PMA-differentiated HL60 released O2- upon stimulation with opsonized crocidolite.

Asbestos↗
PubMed1993

Tolerance of CD8+ T cells developing in parent-->F1 chimeras prepared with supralethal irradiation: step-wise induction of tolerance in the intrathymic and extrathymic environments.

Tolerance of CD8+ cells was examined in parent-->F1 bone marrow chimeras (BMC) prepared with supralethal irradiation; host class I expression in the chimeras was limited to non-BM-derived cells. In terms of helper-independent proliferative responses in vitro and induction of graft-vs.-host disease on adoptive transfer, CD8+ cells from long-term chimeras showed profound tolerance to host antigens irrespective of whether the cells were prepared from the thymus or from spleen or lymph nodes. By limiting dilution analysis, cytotoxic T lymphocyte (CTL) precursors specific for host antigens were rare in the extrathymic lymphoid tissues. In the thymus, by contrast, host-specific CTL precursors were only slightly less frequent than in normal parental strain mice. These host-specific CD8+ cells survived when BMC thymocytes were transferred intravenously to a neutral environment, i.e., to donor strain mice. When transferred to further BMC hosts, however, most of the host-reactive cells disappeared. Collectively, the data suggest that tolerance of CD8+ cells in BMC hosts occurs in both the intrathymic and extrathymic environments. In the thymus, contact with host antigens on thymic epithelial cells deletes CD8+ cells controlling helper-independent proliferative responses and in vivo effector functions but spares typical helper-dependent CTL precursors. After export from the thymus, most of the CTL precursors are eliminated after contacting host antigens on stromal cells in the extrathymic environment.

Animals↗
PubMed1993

Effects of lead exposure on neurophysiological parameters.

To clarify the chronic effect of lead exposure on the central and peripheral nervous systems (CNS and PNS), we performed neurophysiological tests on 41 lead-exposed male workers. Unexposed workers (controls, N = 39) were examined for auditory brain stem response (ABR), and their ABR parameters were compared with those of 15 lead-exposed workers age-matched to the controls. Neurophysiological tests included those of motor and sensory nerve conduction velocity of the radial nerve (MCV, SCVwa and SCVfw), electroretinograms, pattern reversal visual evoked potential (VEP), ABR, and short-latency somatosensory evoked potentials (SLSEP). Neurophysiological parameters were analyzed by regression analysis [independent parameters: age, exposure duration, and current and time-weighted average lead concentration in whole blood (PbB and TWA-PbB)]. ABR parameters were also tested by Student's t test. Significant negative correlations were found between radial MCV and TWA-PbB and SCVwa and PbB, while significant positive correlations were found between the latency of component N145 of VEP and exposure duration and between the latency of component N20 of SLSEP and PbB. The mean of interpeak latency between component III and V of ABR of 15 lead-exposed workers was significantly prolonged compared with that of the control group. These results suggested that lead exposure has a greater effect on the conduction function in the PNS than in that of the CNS in somatosensory and auditory pathways, and inversely in visual pathway.

Adult↗
PubMed1993

Intrathymic and extrathymic tolerance in bone marrow chimeras.

Parent-->F1 bone marrow (BM) chimeras provide a useful model for studying self tolerance induction. When prepared with supralethal irradiation (1300 cGy) and conditioned with anti-T cell antibodies, parent-->F1 BM chimeras are devoid of host BM-derived cells; host H-2 expression is apparent in both the intrathymic and extrathymic environments but is limited to non BM-derived cells. When parent-->F1 chimeras are injected with T cells from normal parental strain mice, the expression of host H-2 antigens on nonprofessional APC might be expected to induce tolerance through induction of clonal anergy. In practice, this does not occur. Instead, a small proportion of the injected T cells is induced to proliferate and differentiate into effector cells. Tolerance is not seen. Similarly, tolerance is not apparent when thymectomized parent-->F1 chimeras are given parental strain thymus grafts. These findings suggest that the expression of host H-2 antigens in the post-thymic environment of chimeras is not intrinsically tolerogenic for mature T cells or recent thymic emigrants. Interestingly, post-thymic tolerance does occur when parental strain T cells differentiate in the endogenous thymus of chimeras. Thus, when mature CD8+ cells are prepared from thymus vs lymph nodes (LN) of parent-->F1 chimeras, tolerance to host class I antigens is more marked in LN than thymus; this applies to cytotoxic T lymphocyte (CTL) precursors, generated by limiting dilution analysis. It would appear therefore that many of the host-reactive CTL precursors generated in the thymus of chimeras undergo tolerance induction (deletion or irreversible inactivation) in the post-thymic environment. We suggest that such tolerance is a reflection of a covert form of tolerance induced in the thymus: intrathymic contact with host antigens on thymic epithelial cells (TEC) in chimeras does not delete typical CTL precursors, but these cells are rendered "semi-tolerant". When cultured in vitro in the presence of lymphokines, the cells are able to recover and differentiate into CTL. In vivo, however, the cells recognize antigen in the periphery in the relative absence of lymphokines and the cells die. Although host class I expression on TEC in chimeras deletes only a small proportion of CTL precursors, contact with TEC induces strong tolerance of CD8+ cells in terms of helper-independent proliferative responses in vitro and induction of lethal graft-versus-host disease in vivo. We postulate that these latter responses are controlled by high-affinity T cells, whereas typical CTL generated in LDA are predominantly low-affinity cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1993

Measurement of redox states of mitochondrial cytochrome aa3 in regions of liver lobule by reflectance microspectroscopy.

To evaluate the sublobular distribution of oxygen tension, the redox states of mitochondrial cytochrome aa3 were assessed in local regions of the liver lobule by reflectance microspectroscopy. The reflected light from two focused microspots (20 microns in diam) on the surface of the perfused liver, placed on the stage of microscope, was conducted to the spectrophotometer with two separate light guides, giving the reflectance spectra. The redox state of cytochrome aa3 was calculated from the difference in reflectance between 603 and 630 nm. The degree of cytochrome reduction increased with the stepwise decline of the influent or effluent oxygen concentrations. The local oxygen concentration in periportal region for the half-maximal reduction of cytochrome aa3 was higher than that in pericentral region. In addition, the degree of cytochrome aa3 reduction increased with the distance from the periportal region, reflecting a physiological gradient of oxygen tension along the sinusoidal flow, although with an intersinusoidal heterogeneity within liver lobule.

Animals↗
PubMed1993

Renal hypouricemia with both drug-insensitive secretion and defective reabsorption of urate: a novel type of renal hypouricemia.

Renal handling of urate in the hypouricemic patient with increase in both urate clearance (Cur) and Cur/creatinine clearance (Ccr) and normal urinary excretion of urate was studied according to the pharmacological evaluation. In the present case there was no response of urate excretion to either pyrazinamide or probenecid. Both furosemide and prednisolone could not alter Cur and Cur/Ccr. Administration of inosine could have increased Cur, which was greater than Ccr. These results suggest that the present case had the defect of both pre- and postsecretory reabsorption of urate, accompanied by the existence of drug-insensitive secretion of urate, which is different from hitherto known types of renal hypouricemia, i.e. a novel type of renal hypouricemia.

Adult↗