Biomedical subjects

A Tsuji

Publications and source records attributed to A Tsuji.

At least 217 records · Page 12Linked to original sources

PubMed1995

cDNA cloning and functional characterization of rat intestinal monocarboxylate transporter.

A cDNA clone which encodes a monocarboxylate transporter (ratMCT1) was isolated from a rat small intestinal cDNA library, which was screened by using full-length MCT1 cDNA of Chinese hamster ovary cells. The ratMCT1 cDNA was sequenced and predicted a protein of 494 amino acids with twelve potential transmembrane domains. The amino acid sequence showed 93.1% and 84.6% identity to the hamster and human monocarboxylate transporters, respectively. When expressed in Xenopus laevis oocytes, the ratMCT1 cRNA caused a significant increase in the uptake of radiolabeled lactic acid. Poly(A)+ RNA transcripts hybridizing to the ratMCT1 cDNA were detected in rat brain, heart, kidney, lung, muscle and brain capillaries. These results indicate that MCT1 contributes to pH-dependent and carrier-mediated transport of monocarboxylic acids in many tissues, not just in the small intestine.

Amino Acid Sequence↗

PubMed1995

Na(+)- and Cl(-)-dependent transport of taurine at the blood-brain barrier.

The characteristics of carrier-mediated transport of taurine at the blood-brain barrier (BBB) were studied by using primary cultured bovine brain capillary endothelial cells (BCECs), in situ brain perfusion and brain capillary depletion methods in rats. The uptake of [3H]taurine by cultured cells showed that the active transporter functions on both the luminal and antiluminal membranes of BCECs. The kinetic parameters for the saturable transport of taurine were estimated to be: for the luminal uptake, the Michaelis constant, Kt, was 12.1 +/- 0.5 microM, and the maximum uptake rate, Jmax, was 4.32 +/- 0.05 nmol/30 min/mg protein; for the antiluminal uptake, Kt was 13.6 +/- 2.4 microM and Jmax was 2.81 +/- 0.22 nmol/30 min/mg protein. The luminal and antiluminal uptakes of [3H]taurine were each dependent on both Na+ Cl-. Stoichiometric analyses suggest that two Na+ and one Cl- are associated with the luminal uptake of one taurine molecule. beta-Amino acids such as beta-alanine and hypotaurine strongly inhibited the uptake of [3H]taurine, whereas alpha- and gamma-amino acids had little or no effect. Furthermore, by in situ brain perfusion and in vivo brain capillary depletion methods, the carrier-mediated transport found by in vitro experiments was confirmed to function for the translocation of the taurine molecule from the vascular space into the brain. From these results, it was concluded that a Na+ and Cl- gradient-dependent transport (uptake) system for taurine exists in both the luminal and the antiluminal membranes of BCECs.

Amino Acids↗

PubMed1995

Participation of a proton-cotransporter, MCT1, in the intestinal transport of monocarboxylic acids.

A molecular mechanism for the intestinal monocarboxylic acid transport was characterized by using a proton/monocarboxylate transporter, MCT1, in Chinese hamster ovary (CHO) cells, first found by Garcia et al. (Cell, 76, 865-873, 1994). Northern blotting analysis showed that MCT1-isomers exist in the rat and rabbit intestinal enterocytes and Caco-2 cells. The expression of [14C]lactic acid uptake by Xenopus laevis oocytes injected with rabbit intestinal mRNA was reduced by hybridizing the mRNA with a MCT1 cDNA of CHO cells before microinjection used as the antisense DNA. [14C]Lactic acid uptake by CHO cells was pH dependent, saturable, stereospecific, and reduced in the presence of acetic acid, benzoic acid, S- and R-ibuprofen, S- and R-mandelic acid, nicotinic acid, pravastatin, propionic acid and valproic acid. In addition, several monocarboxylic acids were transported in pH-dependent and saturable manners. These results suggest that the intestinal MCT1-related protein contributes to a carrier-mediated absorption for organic weak acid compounds.

Animals↗

PubMed1995

Species differences in hydrolysis of isocarbacyclin methyl ester (TEI-9090) by blood esterases.

Species differences in the hydrolysis of isocarbacyclin methyl ester (TEI-9090) in whole blood and in its separated components were studied in rats, dogs and human. Esterase activity in rat whole blood was approximately 100 and 400 times higher than that in dog and human whole blood, respectively, and was attributed to high plasma activity. In contrast, TEI-9090 hydrolysis activities in dog and human blood were due to red blood cells (RBC), whose activity in humans was slightly suppressed by albumin. In dogs, activity in RBC membranes was 10 times greater than in the cytosol, while in human membrane and cytosol activity was virtually the same. The effects of the esterase inhibitor diisopropylfluorophosphate, bis-p-nitrophenylphosphate (BNPP), eserine, 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) and p-chloromercuribenzoate showed that the rat plasma and RBC cytosol esterases hydrolysing TEI-9090 were carboxylesterase (CarbE) and arylesterase (ArE), respectively. The esterases in dog plasma and RBC membrane were CarbE, and RBC cytosol esterase was ArE. In humans, the esterase activities in plasma, RBC membrane and cytosol were butyrylcholinesterase, CarbE and ArE, respectively.

Animals↗

PubMed1995

In vivo and in vitro evidence for ATP-dependency of P-glycoprotein-mediated efflux of doxorubicin at the blood-brain barrier.

We investigated the role of ATP in the active efflux of doxorubicin (DOX) mediated by P-glycoprotein (P-gp), the multidrug-resistance (MDR) gene product, at the blood-brain barrier. In transient brain ischemic rats prepared with 4-vessel occlusion of vertebral and common carotid arteries for 20 min, a procedure that depleted their brain ATP content to 3% that of normal rats, the estimated permeability coefficient of DOX was increased 17-fold (to 243 +/- 2.5 microL/min/g brain). When the ATP content recovered to a normal level by means of 30-min and 24-hr cerebral recirculation of blood, the permeability coefficient recovered to 14.0 +/- 5.0 and 18.4 +/- 2.3 microL/min/g brain (mean +/- SEM, N = 3-6), respectively, very close to the control permeability (14.3 +/- 1.5 microL/min/g brain). The uptake of DOX by primary cultured brain capillary endothelial cells expressing P-gp at the luminal membrane was increased significantly (up to 2-fold), which correlated well with the decrease of cellular ATP contents caused by treating the cells with metabolic inhibitors. Evidence for the ATP-dependent transport of DOX obtained from the present in vivo and in vitro studies strongly indicates that P-gp in the brain capillaries functions actively as an efflux pump in the physiological state, providing a major mechanism to restrict the transfer of DOX into the brain.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1995

Comparison of casual blood pressure and twenty-four-hour ambulatory blood pressure in high school students.

OBJECTIVE: To perform ambulatory blood pressure monitoring (ABPM) in high school students and to compare the 24-hour values with casual blood pressure (BP). METHODS: Ambulatory BP monitoring was carried out in 190 high school students, 15 or 16 years of age, with elevated casual BP during the first examination at school for hypertension; 66 had elevated casual BP at both the first and the second examinations (group H1), and 124 students were normotensive at the second examination (group H2). Sixty-two students who were consistently normotensive served as control subjects (group N). Average BPs for the 24-hour period, the awake period (10 AM to 10 PM), and the sleeping period (1 AM to 6 AM) were calculated. RESULTS: Average BP for each period was significantly higher in group H1 than in group N. Average BP in group H2 for these periods was intermediate between those of group H1 and of group N. The casual BP had a significant positive correlation with the average BP during the awake period (r = 0.55, p < 0.01 for systolic BP; r = 0.37, p < 0.01 for diastolic BP). In most subjects the casual BP readings were higher than BP during ABPM. CONCLUSIONS: Casual BP measurements correlated with 24-hour BP, but they tended to be higher than the measurements obtained during 24-hour ABPM.

Adolescent↗

PubMed1995

Hepatic uptake of octreotide, a long-acting somatostatin analogue, via a bile acid transport system.

The hepatic transport mechanism of octreotide (Sandostatin), a somatostatin analogue, was studied using freshly prepared rat hepatocytes. The initial uptake rate of octreotide represented exclusively a saturable transport process. The half-saturation constant, Kt, and the maximum uptake-rate, Jmax, for the uptake of octreotide were 91.1 +/- 28.4 microM and 104.6 +/- 19.7 pmol/mg protein/min, respectively. An energy requirement was demonstrated for [14C]octreotide uptake since metabolic inhibitors (DNP, rotenone, antimycin and NaCN) significantly reduced the initial uptake rate. [14C]octreotide uptake was also significantly inhibited by ouabain. [14C]octreotide uptake was reduced in the absence of Na+ in the uptake medium. [14C]octreotide uptake was significantly inhibited by bile acids, iodipamide, d-tubocurarine, whereas it was not inhibited by bilirubin, TEMA and insulin. Competitive inhibition of taurocholic acid was observed for octreotide uptake with the inhibition constant, Ki, of 82 +/- 17 microM. Moreover, a significant inhibitory effect of octreotide was observed for the Na+ dependent uptake of [14C]taurocholic acid. These results suggest that octreotide is transported into hepatocytes via a bile acid carrier-mediated system.

Amino Acid Sequence↗

PubMed1995

Proton-cotransport of pravastatin across intestinal brush-border membrane.

PURPOSE: The purpose of the present study is to clarify the intestinal brush-border transport mechanism of a weak organic acid, pravastatin, an HMG-CoA reductase inhibitor. METHODS: The transport of pravastatin was studied by using intestinal brush-border membrane vesicles prepared from rabbit jejunum, and uptake by the membrane vesicles was measured using rapid filtration technique. RESULTS: The initial uptake of [14C]pravastatin was markedly increased with decreases in extravesicular pH and showed a clear overshoot phenomenon in the presence of a proton gradient (pHin/out = 7.5/5.5). A protonophore, carbonylcyanide p-trifluoromethoxyphenylhydrazone, significantly reduced the uptake of [14C]pravastatin. In addition, an ionophore for sodium, potassium and proton, nigericin, stimulated the uptake of [14C]pravastatin in the presence of a potassium gradient ([K+]in/[K+]out = 0/145 mM). On the other hand, neither the imposition of an inwardly directed sodium gradient nor an outwardly directed bicarbonate gradient stimulated the uptake of [14C]pravastatin. In the presence of a proton gradient (pHin/out = 7.5/5.5), the initial uptake of pravastatin was saturable with the apparent Kt of 15.2 +/- 3.2 mM and Jmax of 10.6 +/- 1.21 nmol/mg protein/10 sec. The uptake of pravastatin was significantly inhibited by monocarboxylic acid compounds such as acetic acid and nicotinic acid in a competitive manner but not by di- or tricarboxylic acids, or acidic amino acid. CONCLUSION: It was concluded that a pH-dependent transport of pravastatin across the brush-border membrane occurs by a proton-gradient dependent carrier-mediated mechanism rather than by simple diffusion of its unionized form.

Animals↗

PubMed1995

Correlation of Tc-99m GSA hepatic studies with biopsies in patients with chronic active hepatitis.

To determine whether scintigraphic findings of Tc-99m DTPA-galactosyl-HSA (GSA) correspond to histopathologic findings, Tc-99m GSA hepatic scintigraphy and biopsy were compared in 65 patients with chronic active hepatitis. After injecting 185 MBq of Tc-99m GSA, anterior images were obtained at 5 minutes and 15 minutes. Scintigrams were classified into three grades according to the extent of visualization of the cardiac blood pool on 5 minute and 15 minute images. Biopsies were subjectively graded for findings of necrosis and fibrosis. Scintigraphic grades on 5 minute images were correlated with hepatic necrosis and fibrosis and those on 15-minute images with hepatic fibrosis. Scintigraphic abnormalities of Tc-99m GSA correlated well with histopathologic abnormalities, especially with hepatic fibrosis and necrosis in patients with chronic active hepatitis.

Adult↗

PubMed1995

Distribution of the Kexin family proteases in pancreatic islets: PACE4C is specifically expressed in B cells of pancreatic islets.

The distribution of Kexin family proteases in adult rat pancreatic islets was investigated by immunohistochemical means using a series of specific antibodies specific for PC1, PC2, PC6, Furin, PACE4A and a recently identified member of the Kexin family, PACE4C. PACE4C expression was limited to B cells of the pancreatic islets. PC2 was found in A and in some D cells more than in B cells and PC1 was evident only in B cells. Furin and PC6 were weakly and evenly expressed in the entire islet. PACE4A was hardly found in the islets. These findings indicated that individual Kexin family proteases are uniquely distributed in the islets and suggested that these proteases share roles in these cells as follows: PC2 is involved in the peptide hormone precursor processing in A cells and in D cells, and PACE4C, PC1 and PC2 (mainly PACE4C and PC1) are responsible for the processing event(s) specific to B cells.

Animals↗

PubMed1995

[Preparation of specific antiserum of secretin and its use in time-resolved fluoroimmunoassay].

We have prepared a specific anti-secretin antiserum and developed a sensitive time-relevant fluoroimmunoassay (TR-FIA) for secretin. An anti-secretin antiserum was produced in rabbit by immunization with synthetic secretin C-terminal 8 residue conjugate to chicken serum albumin as antigen. The TR-FIA was based on the competition between free secretin and biotinylated secretin as a tracer for rabbit anti-secretin antibodies on a goat anti-rabbit IgG antibody coated microtiter plate. After separation of bound and free fractions, a solution of europium (III) ion chelate labeled streptavidin (Eu-SA) was added to the biotinylated secretin bound on the microtiter plate. The microtiter plate was washed and then Eu-SA activity was measured by time-resolved fluorometry. In this TR-FIA, the measurable range of secretin was from 2.5 to 100 pg/assay and the specificity was less than 0.01% against other peptide hormones. For the measurement of secretin in the human plasma, a sample was required for the pretreatment in order to separate from interfering substances. The mean recovery of secretin using commercially reverse phase column was 80.2% (n = 11). The plasma secretin concentrations of normal subjects and those with various diseases could be measured by this proposed TR-FIA.

Animals↗

PubMed1995

Long-term follow-up of idiopathic ventricular arrhythmias in otherwise normal children.

Clinical characteristics and long-term prognosis of 163 children with ventricular arrhythmias without underlying heart diseases (78 with ventricular premature contractions (VPC group), 39 with ventricular couplets (CPLT group), and 46 with ventricular tachycardia (VT group)) were studied by Holter electrocardiographic monitoring (Holter ECG) and treadmill exercise testing. The age of the subjects at the initial examination was 8.9 +/- 3.4 years in the VPC group, 9.9 +/- 3.5 years in CPLT group, and 9.4 +/- 3.1 years in the VT group. The duration of the follow-up was 71.7 +/- 32.1 months in the VPC group, 65.9 +/- 32.8 months in the CPLT group, and 84.0 +/- 31.9 months in the VT group. VPC's disappeared during the follow-up period in 22 (28%) of the 78 children in the VPC group. CPLT's disappeared in 15 (38%) and VPC's disappeared in 9 (23%) of the 39 children in the CPLT group. In the 46 children in the VT group, VT disappeared in 30 (65%), and VPC's disappeared in 17 (37%). One child (2%) in the VT group died of heart failure due to drug-resistant sustained VT. The mean time until the disappearance of VPC's in the 163 patients was estimated to be 115.2 +/- 4.3 months. The mean time until the disappearance of VT in the 46 children in the VT group was estimated to be 89.0 +/- 4.9 months. Multivariate analysis of prognostic factors related to the disappearance of VPC's indicated that nighttime VPC's were significantly more likely to disappear (p = 0.018), and that symptomatic VT was significantly more likely to disappear than asymptomatic VT (p = 0.032), probably because more symptomatic cases received antiarrhythmic therapy. Ventricular arrhythmias in children without underlying diseases often disappeared, and the prognosis was generally favorable. However, appropriate treatment and follow-up were required in children with sustained VT, symptomatic VT, or VT with a high rate of VT.

Adolescent↗

PubMed1995

A new spontaneous allele at the pink-eyed dilution (p) locus discovered in Mus musculus castaneus.

Mutant mice characterized by a cream coat and pink eyes were spontaneously discovered among the descendants of Indonesian wild mice (Mus musculus castaneus). This mutant phenotype was controlled by a single autosomal recessive gene that was allelic to the pink-eyed dilution (p) gene. The mutant mouse phenotypically resembled the original p mouse which was the first mutant identified at this locus. Nevertheless, these two alleles differed in origin, a previous report suggesting that the original p allele was derived from Japanese wild mice (M. m. molossinus). Thus the symbol pcas (pink-eyed castaneus) was proposed for the present mutation allele.

Alleles↗

PubMed1995

Immunoblastic lymphadenopathy-like T-cell lymphoma displaying rearrangement of both IgH and TCR beta genes after 4-year follow-up of idiopathic eosinophilia.

Immunoblastic lymphadenopathy (IBL)-like T-cell lymphoma (IBL-T) occurred in a 60-year-old female after a 4-year follow-up of idiopathic eosinophilia and upper pharyngeal inflammatory tumor with infiltration of mature eosinophils. Gene analysis of tumor cells revealed rearrangement of both IgH and TCR beta genes. The patient died of lymphoma seven months after the onset of the illness, in spite of chemotherapy against lymphoma. The relationship between eosinophilia and the pathogenesis of IBL-T, as well as the significance of the rearrangement of both IgH and TCR beta genes are discussed.

Eosinophilia↗

PubMed1995

Ruptured aneurysm of the distal posterior inferior cerebellar artery in a neonate--case report.

A 34-day-old male presented with a rare neonatal ruptured aneurysm of the distal posterior inferior cerebellar artery manifesting as a 10-day history of enlargement of head circumference. Magnetic resonance imaging revealed hydrocephalus and a round infratentorial enhanced lesion which compressed the medulla. Left vertebral angiography demonstrated an aneurysm on the telovelotonsillar segment of the left posterior inferior cerebellar artery. Ventriculoperitoneal shunt emplacement and proximal artery clipping were performed. The cerebrospinal fluid was bloody, suggesting aneurysm rupture had caused hydrocephalus. His postoperative course was uneventful, and neurological and developmental findings were normal 7 months later. Present neuroimaging, surgical, and neuro-anesthesiology techniques allow successful surgical intervention in cases of neonatal ruptured aneurysm.

Aneurysm, Ruptured↗

PubMed1995

[The advantage of a thin flexible ureteroscope with a new real-time processing system].

To observe the upper urinary tract more clearly with a thin flexible endoscope, we developed a novel real-time processing system. This system enhances the parts of the dark cladding around the core by individual strong brightness of each image fiber. The upper urinary tract in eight patients was examined using a thin flexible endoscope with this system at retrograde pyelography or at urological operation. Six patients had hydronephrosis and 2 had filling defects in the renal pelvis or calyx. The endoscope was 0.75 mm in diameter and had 300 pixels of image fiber. The objective portion could be assessed successfully in seven of eight. Significantly finer and colorful image could be obtained with this system compared with the conventional method and the epithelium could be clearly observed. We conclude that this real-time processing system remarkably improves the quality of image obtained with the thin flexible ureteroscope and makes it a more promising modality in the diagnosis and treatment of the upper urinary tract.

Adult↗

PubMed1995

[Expression of GP160 mRNA in renal cell carcinoma: an attempt to predict the sensitivity of IFN-alpha therapy].

BACKGROUND: Alpha-interferon (IFN-alpha) has significant antitumor activity against advanced renal cell carcinoma (RC). Although numerous clinical trials reported major responses in approximately 15% of patients subjected to INF-alpha therapy, it has been impossible to predict which patients will respond to IFN-alpha. RC cell lines could be subdivided into those that are inhibited by IFN-alpha and those that are not. The expression of GP160 (Aminopeptidase a/kidney-associated differentiation antigens) correlated with resistance to IFN-alpha. In RC xenografts using a mouse model, though GP160(+) RC cell lines were resistant to IFN-alpha, GP160(-) RC cell lines were markedly sensitive to IFN-alpha. METHOD: In this study, we assayed 26 RC patients for the expression of GP160 in mRNA level using RT-PCR (reverse transcription polymerase chain reaction). RESULTS & CONCLUSION: In 23 (88.5%) patients GP160 mRNA expression are remarkable as in normal kidney tissue, but in 2 (7.7%) patients, GP160 mRNA expression was weak. And in 1 (3.8%) patient, GP160 mRNA expression was hardly detectable, and in this patient complete remission was achieved with IFN-alpha therapy for advanced lung and mediastinal metastatic RC.

Adult↗

PubMed1995

Functional expression of transporter for beta-lactam antibiotics and dipeptides in Xenopus laevis oocytes injected with messenger RNA from human, rat and rabbit small intestines.

A heterologous gene expression system, Xenopus laevis oocytes, was used to prove the intestinal absorption of various beta-lactam antibiotics mediated by an H(+)-dipeptide cotransport system in rat, rabbit and human small intestines. The microinjection of mRNA (messenger RNA) from rat intestine into Xenopus laevis oocytes led to significantly higher uptakes of p.o. active cephalosporins including zwitter-ionic derivatives (cephalexin, cephradine and cefadroxil) and dianionic derivatives (cefixime and ceftibuten) in comparison with oocytes injected with water, whereas the uptake of cefazolin, a parenterally administered derivative, was negligible in both mRNA- and water-injected oocytes. The uptake of cefadroxil was reduced significantly in the presence of dipeptide and various beta-lactam antibiotics, but not in the presence of an amino acid. After sucrose density gradient centrifugation of mRNA, the highest expression of transport activities of both cefadroxil and ceftibuten was observed in the same mRNA fraction with a size of 2.20 to 3.75 kilobases. mRNA-injected oocytes showed a marked pH-dependency in the uptakes of cefadroxil and ceftibuten, whereas water-injected oocytes exhibited only modes uptakes. The most stimulated uptakes of cefadroxil and ceftibuten were observed at an external pH of 5.5 and 5.0, respectively. Furthermore, injection of mRNA isolated from either rat rabbit or human small intestine into oocytes produced significantly higher uptake of cefadroxil and ceftibuten compared with those by oocytes injected with water. Thus, intestinal absorption of p.o. active beta-lactam antibiotics was confirmed to be mediated by an H+ gradient-dependent transport system across the brush-border membrane of rats, rabbits and humans. The carrier-protein for this process is likely a dipeptide transport system.

Animals↗