Biomedical subjects

R Hori

Publications and source records attributed to R Hori.

At least 73 records · Page 4Linked to original sources

PubMed1994

Human P-glycoprotein as a multi-drug transporter analyzed by using transepithelial transport system.

To analyze the mechanism of drug transport, mechanism of inhibitors, and physiological substrates of human P-glycoprotein, we established a transepithelial transport system by introducing MDR1 cDNA into LLC-PK1, a pig kidney epithelial cell line. P-glycoprotein functions as a steroid transporter as well as a drug transporter as physiological functions. P-glycoprotein also transports MDR modulators such as cyclosporin A, FK506, and calcium channel blockers.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1994

p-Aminohippurate transport in apical and basolateral membranes of the OK kidney epithelial cells.

p-Aminohippurate (PAH) transport in apical and basolateral membranes of OK cells was studied to characterize each membrane process and to determine which process is important for the vectorial transport of PAH across the OK cell monolayers. PAH uptake from the basal side into OK cells was much higher than from the apical side, and was inhibited almost completely by anion transport inhibitors (probenecid and furosemide) and low temperature. On the other hand, PAH efflux from OK cells to the apical side was higher than to the basal side. The efflux across the apical membrane also was inhibited by the transport inhibitors and low temperature. The uptake of PAH across the basolateral membrane was inhibited by dicarboxylates such as alpha-ketoglutarate, and kinetic analysis showed that the inhibition was a mixed-type. These findings suggest that PAH transport in apical and basolateral membranes of OK cells is a specifically mediated process, and both processes contribute to the vectorial transport of PAH across the cell monolayers. PAH/dicarboxylate exchange may be involved in PAH transport in the basolateral membrane of OK cells.

Animals↗

PubMed1994

Transport of gentamicin and fluid-phase endocytosis markers in the LLC-PK1 kidney epithelial cell line.

In order to characterize the transport of aminoglycoside in the cultured kidney epithelial cell line LLC-PK1, cellular uptake of gentamicin was studied in comparison with those of fluid-phase endocytosis markers, lucifer yellow and horseradish peroxidase. The uptake of gentamicin and fluid-phase markers were time-dependent and were reduced at low temperature. The rate of gentamicin uptake was, however, faster than those of lucifer yellow and horseradish peroxidase, and was saturable. The accumulation of gentamicin in the cells was increasing even after 6 days of incubation, whereas that of lucifer yellow reached a steady state by 1 day. Release of intracellular gentamicin from LLC-PK1 cells was much slower than that of lucifer yellow. Gentamicin uptake, but not lucifer yellow uptake, was increased by reducing the ambient ionic strength. Under the low ionic strength conditions, gentamicin uptake was inhibited by the presence of other aminoglycosides, lysozyme, polyamines such as spermine and inorganic cations such as calcium. These results indicate that gentamicin electrostatically binds to the cell apical membrane and is subsequently taken up by an adsorptive endocytosis in LLC-PK1 cells.

Animals↗

PubMed1993

Transport of organic cation in renal brush-border membrane from rats with renal ischemic injury.

Transport of tetraethylammonium, an organic cation has been studied using renal brush-border membrane vesicles isolated from rats with ischemic and ischemia-reperfusion injury. H+ gradient-dependent uptake of tetraethylammonium slightly, but significantly, decreased in brush-border membrane vesicles from ischemic kidneys. When the kidney was reperfused after ischemia, the extent of the decrease of tetraethylammonium uptake was much greater than that after ischemia alone. The Vmax value of tetraethylammonium uptake by brush-border membrane vesicles from reperfused kidneys was decreased compared with control, without any change in the Km value. The tetraethylammonium uptake by the vesicles from reperfused kidneys was decreases both in the presence and absence of the outward H+ gradient (driving force). Uptake of D-glucose in renal brush-border membrane vesicles was also decreased by ischemia and again, reperfusion caused a further decrease of the uptake. Reperfusion also induced marked changes in the enrichment and recovery of marker enzymes in the isolated brush-border membrane fraction compared with ischemia. These findings suggest that renal ischemic injury altered the transport properties of tetraethylammonium as well as D-glucose, and that reperfusion after ischemia induced further damages on these functions in the brush-border membrane.

Animals↗

PubMed1993

P-glycoprotein-mediated transcellular transport of MDR-reversing agents.

Understanding of the interactions between P-glycoprotein and multidrug resistance (MDR) reversing agents is important in designing more effective MDR modulators. We examined transcellular transport of several MDR modulators by using a drug-sensitive epithelial cell line, LLC-PK1, and its transformant cell line, LLC-GA5-COL300, which expresses human P-glycoprotein on the apical surface. Basal-to-apical transports of azidopine and diltiazem across the LLC-GA5-COL300 monolayer were increased and apical-to-basal transports were decreased compared to those across the LLC-PK1 monolayer, indicating that P-glycoprotein transports azidopine and diltiazem. Movements of nitrendipine and staurosporine across the epithelial monolayer were not affected by P-glycoprotein. These results suggests that some MDR modulators exert their inhibitory effect not only by blocking the initial binding of anticancer drugs but throughout the course of the transport process.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1993

Transport of bestatin in rat renal brush-border membrane vesicles.

Bestatin [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl-L-leucine] is a dipeptide, comprising L-leucine and an unusual beta-amino acid. We studied its transport mechanism in rat renal brush-border membrane vesicles. Uptake of cephradine, an aminocephalosporin, by isolated brush-border membrane vesicles was trans-stimulated and cis-inhibited by bestatin, indicating that these drugs are transported via the same transport system(s). The uptake of bestatin was trans-stimulated by preloading the vesicles with glycylsarcosine, and was cis-inhibited by substrates for the H+/dipeptide cotransport system. Bestatin inhibited tetraethylammonium (an organic cation) uptake, and bestatin uptake was cis-inhibited by substrates for the H+/organic cation antiport system. In addition, bestatin uptake was stimulated by an outward H+ gradient (the driving force for the H+/organic cation antiport system). These findings suggest that bestatin, in spite of being a dipeptide, is transported via not only the H+/dipeptide cotransport system but also the H+/organic cation antiport system in rat renal brush-border membrane.

Animals↗

PubMed1993

Human P-glycoprotein transports cyclosporin A and FK506.

Cyclosporin A, a cyclic undecapeptide, and FK506 are efficient immunosuppressive agents. They also attract attention as effective P-glycoprotein modulators that inhibit P-glycoprotein from binding to anticancer drugs and overcome multidrug resistance. Cyclosporin A itself interacts with a common binding site of P-glycoprotein to which Vinca alkaloids and verapamil bind. We were interested to determine whether cyclosporin A and FK506 are substrates for P-glycoprotein to transport, and we studied their transcellular transport. In LLC-PK1 cells, derived from porcine kidney proximal tubule and forming a highly polarized epithelium, cyclosporin A was transported in a saturable manner. LLC-GA5-COL300, a transformant cell line derived by transfecting LLC-PK1 with human MDR1 cDNA isolated from normal adrenal gland, expresses P-glycoprotein specifically on the apical surface and shows a typical multidrug-resistant phenotype. LLC-GA5-COL300 cells showed increased transport of cyclosporin A from the basal to the apical side. Kinetic analysis showed that this transport was a typical saturable transport with the calculated apparent Michaelis constant (Kappm) and the maximum flux (Vmax) as 8.4 microM and 2.4 nmol/mg protein/h, respectively. LLC-GA5-COL300 also showed increased transport of FK506 from the basal to the apical side. These results indicate that P-glycoprotein transports the immunosuppressive agents cyclosporin A and FK506.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1993

Characteristics of organic cation transporter in rat renal basolateral membrane.

Characteristics of organic cation transport system were studied in rat renal basolateral membrane and compared with those in brush-border membrane. We first examined the effect of various chemical modifiers on tetraethylammonium uptake by the membrane vesicles. Treatment with N,N'-dicyclohexylcarbodiimide and phenylglyoxal (carboxyl groups and arginine residues specific reagent, respectively) resulted in inhibition of tetraethylammonium transport in both basolateral and brush-border membranes. Tetraethylammonium uptake by brush-border, but not by basolateral, membrane vesicles was decreased by diethyl pyrocarbonate, histidine residues specific reagent, treatment. Treatment of sulfhydryl groups with HgCl2 decreased tetraethylammonium transport in both membranes. However, in contrast to brush-border membrane, unlabeled tetraethylammonium failed to protect against the inhibition of [14C]tetraethylammonium uptake by p-chloromercuribenzene sulfonate in basolateral membrane. We next examined the inhibitory effect of various organic cations on tetraethylammonium uptake. The order of inhibitory potency of organic cations was somewhat different between two membranes. These findings suggest that the characteristics of organic cation transport systems in basolateral and brush-border membranes were different in regard to essential amino acid residues and the affinity of substrates.

Amiloride↗

PubMed1993

Nonlinear mixed effect modeling of the pharmacodynamics of natriuretic peptides in rats.

Natriuretic peptides have not only natriuretic/diuretic but also hypotensive activities, and the decreased renal perfusion caused by the excessive hypotension is known to attenuate the diuretic actions. The present study was designed to examine the relationship between the dosing (intravenous constant infusion) rates and the diuretic actions of alpha-rat atrial natriuretic peptide (alpha-rANP) and rat brain natriuretic peptide (rBNP) in rats, and population (nonlinear mixed effect model) analysis was applied to these complicated diuretic actions. The intrinsic diuretic activities of alpha-rANP and rBNP could be analyzed, and the effects of blood pressure, heart rate, and also inhibition of degradation enzyme on the diuresis of natriuretic peptides were estimated simultaneously. The population analysis was useful for analyzing such pharmacodynamic data for which the individual analysis could not be applied easily.

Animals↗

PubMed1993

Patient-nurse collaboration: a comparison of patients' and nurses' attitudes in Finland, Japan, Norway, and the U.S.A.

While there has been an increasing emphasis on patients' participation in decisions concerning health care and nursing in the literature as an ideal, it is not clear to what extent patients and nurses assume the consumerist attitude regarding health-care decision making. With the view that attitudes people hold regarding their role in health care and nursing will primarily affect the way they behave in health-care situations, a multinational study was carried out to examine five sets of attitudes regarding consumerism held by patients in acute-care hospitals and nurses working in them. The findings from the surveys in Finland, Japan, Norway, and the U.S.A. indicate that while the patients and the nurses in these countries tend to lean toward the consumerist perspective in their attitudes, there were significant differences in the acculturation of these attitudes among the countries and between the patients and nurses. Two different models for the explanation of attitude regarding collaborative decision making in nursing practice emerged for the patients and the nurses as groups. For both groups, however, age and the more general consumerist attitudes have a bearing on their attitudes regarding collaboration in nursing.

Adult↗

PubMed1993

Decreased cellular toxicity of neomycin in a clonal cell line isolated from LLC-PK1.

We have previously shown in LLC-PK1 cells, that apical membrane enzyme activity was inhibited by aminoglycoside antibiotics (Am. J. Physiol. 254, C251-C257, 1988). In the present study, the relationship between the lethal cytotoxic effect of aminoglycoside and its effect on apical membrane enzyme was examined by establishing aminoglycoside resistant cells. A clonal cell line, LLC-PK1/NRa3, was isolated from parent LLC-PK1 cells in the presence of neomycin. Neomycin inhibited colony formation and increased the number of floating dead cells in parent LLC-PK1 cultures. In contrast, these cytotoxic effects of neomycin were negligible or less pronounced in NRa3 cells, indicating that NRa3 cells were more resistant to neomycin compared with the parent cells. The inhibitory effect of neomycin on apical enzyme activity was significantly weaker in NRa3 cells than in the parent cells. These results suggest that a common mechanism is involved in the aminoglycoside-induced reductions in the apical enzyme activity and in cell viability of LLC-PK1 cells.

Alkaline Phosphatase↗

PubMed1993

Continuous assessment of hemodynamic control by complex demodulation of cardiovascular variability.

Usefulness of complex demodulation (CDM) in assessing the frequency components of cardiovascular variability was assessed and, subsequently, this technique was utilized to determine the time-dependent responses of the low-frequency (LF) and high-frequency (HF) amplitudes of heart rate and blood pressure variabilities during postural tilt. CDM provides the time-dependent changes in amplitude of a particular frequency component on a continuous basis. Analysis of simulated data showed that CDM has sufficient frequency resolution to separately measure LF and HF amplitudes with a time resolution < 15 s and that CDM is robust to alterations in the frequency of the components. Analysis of actual data during postural tilt test in 23 young healthy subjects demonstrated that the HF amplitude of heart rate, an index of cardiac parasympathetic tone, rapidly decayed with head-up tilt (P < 0.01) and increased quickly showing an overshoot with tilt back to the supine position (P < 0.01). The LF amplitude of blood pressure, an index of vasomotor sympathetic activity, showed marked rhythmic fluctuation at an interval of 48-100 s during head-up tilt (P < 0.01), synchronizing with similar fluctuation in the LF amplitude of heart rate (P < 0.01). These results suggest that CDM can be used to provide a continuous assessment of cardiovascular variability components and that the dynamic responses of autonomic circulatory control to upright posture result in a phasic modulation of LF amplitude.

Adult↗

PubMed1993

Transport of organic anion in the OK kidney epithelial cell line.

Transport of an organic anion, p-aminohippurate (PAH), was investigated in an established cell line derived from the American opossum kidney (OK). Transepithelial transport and accumulation of PAH were measured in OK cells cultured on permeable supports. Specifically mediated transcellular transport of PAH across OK cell monolayers occurred only from basal to apical side, and the accumulation of PAH in the cells was much larger from basal side than from apical side. Both the transcellular transport of PAH from basal to apical side and the accumulation from basal side were temperature dependent, saturable, and inhibited by organic anions such as probenecid but not by organic cations. In contrast, such a specific transport of PAH was not observed in LLC-PK1, another epithelial cell line derived from pig kidney. These findings suggest that OK cells can transport PAH by the organic anion transport system unidirectionally from basal to apical side across the cell monolayers and that OK cells might be a useful model for the study of organic anion secretion in the renal proximal tubules.

Animals↗

PubMed1993

p-Aminohippurate transport in rat renal brush-border membranes: a potential-sensitive transport system and an anion exchanger.

Transport mechanisms of p-aminohippurate (PAH) were investigated in rat renal brush-border membrane vesicles. The uptake of PAH was stimulated by an inside-positive membrane potential created by K+ and valinomycin. This potential-stimulated uptake of PAH was inhibited by various anion transport inhibitors and was saturable. In addition, PAH uptake in the presence of valinomycin was linearly increased in proportion to log[K+]out/[K+]in. On one hand, PAH uptake was stimulated by [14C]PAH/PAH or [14C]PAH/Cl- exchange, and the [14C]PAH/PAH exchange was insensitive to the membrane potential. The uptake by the exchanger was also inhibited by anion transport inhibitors, but the potential-stimulated uptake of PAH was more sensitive to furosemide and 4,4'-diisothiocyano-2,2'-disulfonic stilbene. On the contrary, [14C]PAH/PAH exchange was more sensitive to urate than the potential-stimulated uptake of PAH. These findings indicate that PAH is transported by two distinct transport systems in rat renal brush-border membranes, a potential-sensitive transport system and an anion exchanger.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed1993

Population pharmacokinetics of theophylline. II: Intravenous infusion to patients with stable chronic airway obstruction.

The population pharmacokinetics of theophylline were studied in 55 patients with stable chronic airway obstruction. Two hundred and seventy six theophylline serum concentrations after intravenous short infusion were analyzed using a nonlinear mixed-effect model. The influence of hepatic dysfunction, smoking habit, age and the measurement of arterial blood gases (oxygen tension: PaO2, carbon dioxide tension: PaCO2, blood pH) and clinical laboratory tests (serum albumin concentration, haematocrit) on the pharmacokinetic parameters of theophylline was examined by the likelihood ratio test. Assessment of each factor was made by a forward selection method. In the final regression model, the total body clearance (CL, l/h/kg) was related to the value of PaCO2 as well as to the presence of hepatic dysfunction, and the volume of distribution (Vd, l/kg) was related with the PaCO2 value as expressed in the following equations: CL = exp(-3.78 - 0.525.HF + 0.0233.PaCO2) and Vd = exp(-1.12 + 0.00934.PaCO2), where HF is a categorical variable with a value of unity if a patient has hepatic dysfunction otherwise zero. The interactions among blood gas measurements were observed and the CL and Vd of theophylline would be inversely correlated with PaO2 or pH, if we selected PaO2 or blood pH to be a more important factor than PaCO2. The inter-individual variabilities in CL and Vd were 38.5% and 12.5%, respectively, and the residual variability in theophylline serum concentrations was 10.6% as a coefficient of variation. This final model and the population parameters of theophylline will be useful for individualization of a drug dosage regimen by means of the Bayesian method.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1993

Population pharmacokinetics of theophylline. I: Intravenous infusion to children in the acute episode of asthma.

The population pharmacokinetics of theophylline during constant-rate intravenous infusion has been studied in 66 children (8.7 +/- 4.7 year of age; 26.0 +/- 12.8 kg, mean +/- S.D.) with an episode of acute asthma. One hundred and twelve theophylline serum concentrations (13.9 +/- 4.8 micrograms/ml) collected retrospectively were analyzed using a nonlinear mixed-effect model. The influence of hepatic dysfunction, age, gender, days after admission, blood gas measurements (PaO2, PaCO2, blood pH) and clinical analyses data (total serum protein, albumin concentration, haemoglobin concentration, haematocrit) on theophylline clearance was examined by the likelihood ratio test. A final estimate of population mean clearance was 58.6 ml/kg/h, which was decreased by 36% in patients with hepatic dysfunction. Other factors tested displayed no statistically significant effect on theophylline clearance. The inter-individual variability in clearance was 26% while the intra-individual variability in theophylline concentrations was 2.6 micrograms/ml as a standard deviation, which was almost double that observed for stable patients. Taking into account that the therapeutic window of this drug is 10-20 micrograms/ml, this value indicates a relatively large intra-individual variability and suggests that frequent (daily) monitoring of serum concentrations is necessary for patients with an episode of acute asthma.

Acute Disease↗

PubMed1993

Transport mechanism of choline in rat renal brush-border membrane.

The transport mechanism of choline was examined using rat renal brush-border membrane vesicles in comparison with tetraethylammonium transport. The stimulatory effect of an outward H+ gradient on choline uptake was weak compared with that on tetraethylammonium uptake. [14C]Tetraethylammonium uptake was cis-inhibited and trans-stimulated by choline, but the effects were less pronounced than those produced by unlabeled tetraethylammonium. [3H]Choline uptake was trans-stimulated by unlabeled choline, but not by tetraethylammonium. An interior-negative membrane potential induced marked stimulation of choline uptake against its concentration gradient (overshoot phenomenon), and the uptake was saturable with an apparent Km of 0.77 mM. Various compounds such as hemicholinium-3 inhibited the choline uptake by renal brush-border membrane vesicles, but a sulfhydryl reagent did not. These findings suggest that choline can be actively transported by a carrier-mediated system driven by cell interior-negative membrane potential in renal brush-border membrane, and this system may play an important role in the tubular reabsorption of choline.

4-Chloromercuribenzenesulfonate↗

PubMed1993

Protective effect of N-acyl amino acids (NAAs) on cephaloridine (CER) nephrotoxicity in rabbits.

The protective effect of N-acyl amino acids (NAAs) against cephaloridine (CER)-induced nephrotoxicity was studied in rabbits. A large single intravenous dose of CER (more than 100 mg/kg) induced severe proximal tubular necrosis. Simultaneous treatment with several NAAs (at dosages of 100, 200 mg/kg, etc., i.v.), such as N-benzoyl-beta-alanine (NBBA), N-benzoyl-6-aminocaproic acid, and N alpha,epsilon-dibenzoyl-D,L-lysine, remarkably suppressed the histopathological damage in the kidney induced by CER. NAAs have generally low toxicity in laboratory animals (e.g., the LD50 of NBBA was more than 3,000 mg/kg, i.v. in rats), and NAAs were suggested to be good candidates for reducing the nephrotoxicity of CER and other beta-lactam antibiotics.

Alanine↗