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Biomedical subjects

R Nishigaki

Publications and source records attributed to R Nishigaki.

At least 37 records · Page 2Linked to original sources
PubMed1998

Evasion of UVC-induced apoptosis by photorepair of cyclobutane pyrimidine dimers.

Cyclobutyl pyrimidine dimer (CPD) photolyase is known to reverse pyrimidine dimers specifically under illumination with visible light. OCP13, a Medaka cell line showing a high level expression of the gene for CPD photolyase, completely reversed pyrimidine dimers induced by 20 J/m2 UVC by 1 h of photorepair. When OCP13 cells were irradiated with 20 J/m2 UVC, morphological changes such as shrinkage of cells, distorted nuclear shape, and decrease in the number of nucleoli appeared 2 to 4 h after UVC irradiation. Thereafter, the irradiated cells began to detach from the substratum, and DNA ladders were observed in the DNA extracted from detached cells. Thus, these changes in cells after UVC exposure were used to characterize the progression of UV-induced apoptosis in OCP13 cells. Although formation of DNA ladders and cell detachment were blocked by cycloheximide treatment prior to UVC exposure, the morphological changes were not. With photorepair treatment, even after the morphological changes appeared cells were still able to restore their normal morphological features and remained attached. On the other hand, the cell-cycle progression in UVC-irradiated cells was arrested even after photorepair of pyrimidine dimers. Thus, photorepair can rescue cells from UV-induced apoptosis, although DNA damage other than that of pyrimidine dimers, as well as additional non-DNA damage, possibly remained, and DNA replication was left inhibited. Among the various kinds of damage induced by UVC irradiation, the presence of pyrimidine dimers is proposed to be the major trigger for UVC-induced apoptosis.

Animals↗
PubMed1998

Sonodynamically-induced cell damage with fluorinated anthracycline derivative, FAD104.

The ultrasonically-induced in vitro cell damaging effect of fluorine-containing anthracycline derivative (FAD104) was investigated. Sarcoma 180 cells suspended in air-saturated PBS were exposed to ultrasound for up to 60 s in the presence and absence of FAD104. The rate of inducing cell damage with ultrasound was doubled with 80 microM FAD104, while no cell damage was observed with FAD104 alone. This enhancement was significantly inhibited by histidine, which may suggest a sonochemical mechanism.

Animals↗
PubMed1998

Antitumor effect sonodynamically induced by focused ultrasound in combination with Ga-porphyrin complex.

The antitumor effect of focused ultrasound in combination with Ga-porphyrin complex, 7,12-bis(1-decyloxyethyl)-Ga(III)-3,8,13,17 tetramethylporphyrin-2,18-dipropionyl diaspartic acid (ATX-70), on the growth of experimental murine tumors was examined. Walker 256 tumors implanted in rat kidneys were exposed in a progressive wave field for 5 min to focused ultrasound at 500 kHz with the second harmonic (at 1 MHz) superimposed after administration of ATX-70 to the rats. A significant antitumor effect was obtained at a focal-spot average acoustic intensity of 12 W/cm2 or higher with an ATX-70 dose of not less than 2.5 mg/kg. When the acoustic intensity was 8 W/cm2 or lower, no significant effect was observed even at an ATX-70 dose of 2.5 mg/kg. Also when the ATX-70 dose was 1.0 mg/kg or lower, no significant effect was observed even at a focal-spot average acoustic intensity of 40 W/cm2.

Animals↗
PubMed1998

Nitric oxide production and energy state in the heart after endotoxin administration.

To evaluate nitric oxide (NO) production and the energy state of the heart after endotoxin administration, Wistar rats were injected i.p. with 10 mg/kg Escherichia coli lipopolysaccharide (endotoxin). Morphologic changes, plasma nitrite concentration, expression of inducible NO synthase (iNOS), and cardiac energy state, as reflected by several metabolites, were observed chronologically 0 (control), 4, 6 and 8 h after endotoxin administration. Electrocardiography (ECG) demonstrated arrhythmia after endotoxin administration. Biochemically, NO production increased in blood and iNOS increased in the heart. The amount of myocardial beta-ATP measured by 31P magnetic resonance spectroscopy (31P-MRS) increased transiently and then decreased. This transient increase might be a hyperdynamic response to endotoxin administration. At 4 and 6 h after endotoxin administration, pH measured by 31P-MRS was slightly decreased, but this decline was not statistically significant. On the other hand, the amount of lactate in heart samples increased in the 1H magnetic resonance spectra (1H-MRS). Ultrastructurally, in cardiovascular tissue, intracytoplasmic organelles were observed to be injured in blood vessels and cardiomyocytes associated with mast cell infiltration. These results suggest significant metabolic and morphologic abnormalities in the heart after endotoxin administration.

Animals↗
PubMed1998

Salicylic acid-induced lipid peroxidation in rat liver microsomes.

Rat liver microsomes (1 mg protein/ml) were incubated at 37 degrees C with non-steroidal anti-inflammatory drug, salicylic acid (5 mM) in the presence of 0.2 mM NADPH. The amounts of thiobarbituric acid reactive-substances (TBARS) were increased remarkably by the incubation for 30 min. TBARS formation was dependent on salicylic acid concentration. Salicylic acid-induced lipid peroxidation was not observed in the absence of NADPH. It was also inhibited by the presence of cytochrome P450 inhibitor, SKF-525A. The singlet oxygen's scavengers such as histidine suppressed salicylic acid-induced lipid peroxidation. These results suggested that salicylic acid-induced lipid peroxidation was coupled with the metabolism through cytochrome P450 in rat liver microsomes. The singlet oxygen may be involved in the reaction.

Animals↗
PubMed1997

Sonodynamically-induced in vitro cell damage enhanced by adriamycin.

Sonodynamically-induced cell damage and active oxygen generation enhanced by adriamycin (ADM) were compared in the same in vitro insonation set-up. Significant enhancement of the rates of both ultrasonically-induced cell damage and nitroxide generation was demonstrated with 40-160 microM ADM. Both rates correlated very well resulting in a correlation coefficient of more than 0.99. The enhancement of both rates was suppressed by 10 mM histidine. These results are consistent with the hypothesis that ultrasonically-generated active oxygen plays a major role in the sonodynamically-induced cell damage enhanced by ADM.

Animals↗
PubMed1997

Methotrexate is excreted into the bile by canalicular multispecific organic anion transporter in rats.

Methotrexate [(+) amethopterin, L-MTX] has two carboxyl groups in its structure and is eliminated mainly by excretion into urine and bile. To investigate the biliary excretion mechanism of L-MTX, we performed in vivo and in vitro studies using mutant rats, Eisai hyperbilirubinemic rats (EHBRs), whose canalicular multispecific organic anion transporter (cMOAT) is defective as a consequence of heredity. After i.v. administration of L-MTX to EHBRs, its plasma disappearance and biliary excretion was slower than in normal Sprague Dawley rat (SDR). ATP-dependence and overshoot phenomena were observed in the uptake of [3H]L-MTX by canalicular membrane vesicles (CMV) prepared from SDR, whereas no ATP-dependence was observed in CMV from EHBRs. The ATP-dependent uptake of L-MTX by SDR CMV exhibited saturable kinetics with a Km of 295 microM. L-MTX competitively inhibited the ATP-dependent uptake of [3H]2,4-dinitrophenyl-S-glutathione, a typical substrate for cMOAT, and the inhibition constant (Ki) of L-MTX was comparable with its own Km. These results suggest that L-MTX is excreted into bile by cMOAT. The inhibitory effects of L-MTX and its optical isomer, (-) amethopterin (D-MTX), on the uptake of [3H]L-MTX differed with Kis of 326 and 93 microM, respectively, indicating that the biologically inactive D form has a higher affinity for cMOAT than L-MTX.

Adenosine Triphosphate↗
PubMed1997

Sonodynamically induced antitumor effect of gallium-porphyrin complex by focused ultrasound on experimental kidney tumor.

The antitumor effect of a gallium-porphyrin complex, ATX-70, induced by focused ultrasound, on colon 26 carcinoma implanted in a mouse kidney was investigated. Colon 26 tumors were exposed to focused ultrasound at 500 kHz and 1 MHz in a progressive wave mode. Both frequency components were superimposed onto each other in the focal zone to efficiently produce cavitation in the tumor. ATX-70 was administered intravenously at the dose of 2.5 mg/kg, 24 h before the ultrasonic exposure. Antitumor effects were evaluated by histological observation 7 days after the exposure. The destruction of tumor tissue was observed with the ultrasonic treatment in combination with ATX-70, while neither the treatment with ATX-70 alone nor that with ultrasound alone caused any necrosis. These results first demonstrated that antitumor effects of a porphyrin compound can be induced by focused ultrasound in a progressive wave mode.

Animals↗
PubMed1997

Differential distribution of ecNOS and iNOS mRNA in rat heart after endotoxin administration.

To evaluate the relationship between cardiovascular injury and the pathological significance of endothelial constitutive nitric oxide synthase (ecNOS) and inducible nitric oxide synthase (iNOS) in endotoxic shock, Wistar rats were injected intraperitoneally with 10 mg/kg Escherichia coli endotoxin and the resulting cardiovascular changes observed using immunohistochemistry, immunoelectron microscopy, the reverse transcription polymerase chain reaction (RT-PCR) and in situ hybridization at 4, 6, 8 and 10 h after endotoxin administration. Immunohistochemical and electron microscopic observations showed that ecNOS was localized in the cytoplasmic vesicles and rough endoplasmic reticulum of the endothelium of coronary arteries and intermyocardial capillaries in both control and endotoxin-treated rats. iNOS was localized in the cytoplasmic vesicles and endoplasmic reticulum of vascular endothelial cells, vascular smooth muscle cells and cardiomyocytes after endotoxin administration. The RT-PCR study confirmed the expression of ecNOS and iNOS mRNA in the heart tissues of all animals including controls. In situ hybridization showed that ecNOS mRNA was expressed in the cytoplasm of vascular endothelial cells in control and endotoxin-treated rats. After endotoxin administration, iNOS mRNA was strongly expressed in vascular endothelial cells, vascular smooth muscle cells, cardiomyocytes and a small number of macrophages. Bacterial lipopoly-saccharide induces rapid release of nitric oxide in the microvasculature and cardiomyocytes resulting in the depression of cardiomyocyte contraction. These findings may describe the cardiac response after endotoxin treatment.

Animals↗
PubMed1997

Biliary excretion of pravastatin in rats: contribution of the excretion pathway mediated by canalicular multispecific organic anion transporter.

The biliary excretion of pravastatin in normal rats and Eisai hyperbiliruminemic rats (EHBRs) was examined in vivo and in vitro using bile canalicular membrane vesicles (CMVs). In vivo, the total body clearances at steady-state (CLtot) for both rat strains decreased as the infusion rate increased. At the lowest infusion rate, CLtot for normal rats was 1.6 times higher than that for EHBRs. Under this set of conditions, the biliary excretion clearance (CLbile), defined as the biliary excretion rate at steady-state divided by the concentration in the liver (Cliver), for normal rats was 3-fold higher than that for EHBRs. The CLbile fell markedly with increasing Cliver for normal rats and the Michaelis constant (KM) for Cliver was 180 microM; in contrast, the degree of saturation was slight if any in EHBRs. In vitro, the uptake of pravastatin by CMVs prepared from normal rats exhibited clear ATP-dependence, whereas only a minimal effect of ATP was observed on the uptake by CMVs from EHBRs. Transport kinetic studies were performed over a wide range of pravastatin concentration (0.2-10,000 microM) with a tracer tritium-labeled pravastatin. Saturation was observed both in the ATP-dependent (KM: 220 microM) and ATP-independent (KM: 480 microM) uptake by CMVs prepared from normal rats. ATP-dependent uptake of 2, 4-dinitrophenyl glutathione, a typical substrate for the canalicular multispecific organic anion transporter (cMOAT), was inhibited by pravastatin in a concentration-dependent manner and the resultant inhibitory constant of pravastatin (170 microM) was comparable with the KM value of ATP-dependent pravastatin uptake itself. In conclusion, biliary excretion of pravastatin is mediated mainly by cMOAT in normal rats. This can explain the decrease in the biliary excretion of pravastatin in EHBRs.

Animals↗
PubMed1996

Sonodynamically induced antitumor effect of pheophorbide a.

The sonodynamically induced antitumor effect of pheophorbide a (Ph-a) was investigated. Both in vitro and in vivo effects on sarcoma 180 were tested in combination with ultrasound at 2 MHz. The rate of ultrasonically induced cell damage in air-saturated suspension was enhanced by twice with 80 microM Ph-a. This enhancement was significantly inhibited by histidine, which may suggest it was mediated by ultrasonically induced oxidation. For mice, 5 mg/kg Ph-a was administered before the insonation, and ultrasound stopped the tumor growth at an intensity with which ultrasound alone showed only a slight antitumor effect.

Animals↗
PubMed1996

Uptake is the rate-limiting step in the overall hepatic elimination of pravastatin at steady-state in rats.

PURPOSE: Of the HMG-CoA reductase inhibitors, the hydrophilic pravastatin has been shown to exhibit relatively specific inhibition of cholesterol synthesis in the liver. As one of the reasons for this relatively specific pharmacological activity, we demonstrated that the tissue distribution of pravastatin is limited because of its high hydrophilicity, while hepatic uptake by active transport takes place at the liver surface via a multispecific anion transporter (M. Yamazaki et al., Am. J. Physiol., 264, G36-44, 1993). In this study, we examined the hepatic elimination of pravastatin at steady-state. METHODS: After i.v. infusion, the plasma concentrations of pravastatin in both arterial and hepatic venous blood were measured. RESULTS: The hepatic availability at steady-state exhibited a clear increase on increasing the infusion rate of pravastatin. The total hepatic elimination rate at steady-state exhibited Michaelis-Menten type saturation with the drug concentration in the capillary defined by typical mathematical models (i.e., well-stirred, parallel-tube and dispersion models), K(m) and Vmax values being comparable with those obtained from analysis of the initial uptake velocity using in vitro isolated hepatocytes. CONCLUSIONS: These results indicate that overall hepatic intrinsic clearance of pravastatin at steady-state is regulated by the uptake process, followed by rapid metabolism and/or biliary excretion with minimal efflux to the circulating blood.

Animals↗
PubMed1996

Membrane lipid peroxidation as a mechanism of sonodynamically induced erythrocyte lysis.

Sonodynamically induced lipid peroxidation with haematoporphyrin (Hp) was studied using rat erythrocytes. Both suspensions of erythrocyte ghosts and of intact erythrocytes were exposed to ultrasound in the same way in the presence and absence of Hp (80 microM). The lipid peroxidation in erythrocyte ghost membranes was estimated by measuring the amount of reactive substance produced from thiobarbituric acid added immediately after the exposure. Haematoporphyrin multiplied the ultrasonically induced lipid peroxidation by three to five times, while Hp alone showed no peroxidation. A 24-h interval between the exposure and the preparation for measurement did not increase the measured amount of peroxide. In the presence of Hp the estimated peroxidation rate and the rate of erythrocyte lysis correlated quite well for each acoustic condition and for each chemical condition such as the presence or absence of active oxygen scavengers in the suspension. The sonodynamically induced lipid peroxidation with Hp was doubled by deuterium oxide substitution for suspension medium and was significantly reduced by histidine, by sodium azide, and also by nitrogen substitution for saturation gas, whereas superoxide dismutase and mannitol showed no significant inhibitory effect. These results are consistent with a hypothesis that lipid peroxidation in membranes by singlet oxygen is the primary mechanism of sonodynamically induced erythrocyte lysis with Hp.

Animals↗
PubMed1996

Sonodynamically induced antitumor effect of a gallium-porphyrin complex, ATX-70.

The sonodynamically induced antitumor effect of a gallium-porphyrin complex, ATX-70, was evaluated in mice bearing colon 26. In order to find the optimum timing for the ultrasonic exposure after the administration of ATX-70, the ATX-70 concentrations in the plasma, skin, and tumor were measured and analyzed. Antitumor effect was estimated by measuring the tumor size. When used alone, ultrasound showed a slight antitumor effect, which became increasingly significant as the dose of ATX-70 was increased, while use of ATX-70 alone had no significant effect. At an ATX-70 dose of 2.5 mg/kg or higher, the average tumor size decreased to smaller than a half by three days after the ultrasonic exposure. This was smaller than a third of the size of the untreated tumors on the same day. From these results, it is concluded that ATX-70 significantly sensitizes tumors to ultrasound, demonstrating a synergistic antitumor effect.

Animals↗
PubMed1995

[Kinetic analysis of hepatobiliary transport of drugs: importance of carrier-mediated transport].

The liver is the major organ involved in the metabolism and elimination of xenobiotics. Evaluating accurately hepatic clearance is very important for predicting the pharmacological effect and/or side-effects of drugs, as well as changes in drug disposition during disease. Recently, for many endogenous and exogenous compounds (including drugs), it has been reported that carrier-mediated transport contributes to hepatic uptake and/or biliary excretion. In particular, primary active transport mechanisms have been shown to be responsible for the biliary excretion of anticancer drugs, endogenous bile acids and organic anions including glutathione and glucuronic acid conjugates. We have found that a rate-limiting step for several drugs in terms of the hepatic clearance was the membrane transport process. For these drugs, such a saturable transport process can be one of the major determinants which influence not only hepatic clearance itself but also disposition (in other words, plasma elimination) in the whole body. We reviewed the carrier-mediated transport mechanisms involved in the hepatic uptake and biliary excretion processes, the multiplicity of transport systems, and further, the physiological meaning of these complex transport systems in the body.

Animals↗
PubMed1994

Sonochemical activation of hematoporphyrin: an ESR study.

The production of 2,2,6,6-tetramethyl-4-piperidone-N-oxyl by reaction of 2,2,6,6-tetramethyl-4-piperidone (TMPone) with ultrasonically generated active species in oxygenated solutions of hematoporphyrin (Hp) was studied by electron spin resonance spectroscopy. The nitroxide production rate in air-saturated TMPone solutions in phosphate-buffered saline of pH 9.0 was significantly higher in the presence of Hp than in its absence. The enhancement of nitroxide production by Hp was significantly inhibited in the presence of sodium azide or histidine in the solution. The production rate with Hp was doubled by substitution of deuterium oxide, while the rate without Hp increased only modestly. These results suggest that a substantial amount of active oxygen can be generated by ultrasound in aqueous solutions of Hp. Since the production rate was not reduced by mannitol and no nitroxide was produced in nitrogen-saturated solutions, it appears that hydroxyl radicals do not account for a major portion of the active oxygen species which reacted with TMPone to yield a nitroxide.

Electron Spin Resonance Spectroscopy↗