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Biomedical subjects

K Inui

Publications and source records attributed to K Inui.

At least 181 records · Page 10Linked to original sources
PubMed1997

Effects of beta mercaptoethanol on the proliferation and differentiation of human osteoprogenitor cells.

Antioxidants are known to influence metabolism and promote cell survival in a number of cell culture systems. However, their effects on the modulation of bone cell differentiation in vitro are not clearly defined. In the present studies we have investigated the effects of beta-mercaptoethanol (beta ME) and ascorbate alone and in combination on human osteoprogenitors derived from bone marrow fibroblasts. In primary marrow cultures, beta ME stimulated colony formation (2-fold), alkaline phosphatase activity (3.5-fold) and, increased DNA synthesis (8-fold) after 21 days. Cell proliferation was increased significantly by beta ME during the first 4 days of a 10-day culture period, indicating stimulation of marrow osteoprogenitor proliferation. Ascorbate did not significantly augment the effects of beta ME in primary cultures or long-term cultures of passaged bone marrow fibroblasts. These findings indicate a potential beneficial role for beta ME addition for the optimal maintenance of colony formation, cell proliferation and differentiation of marrow osteoprogenitor cells in primary human bone marrow fibroblast cultures.

Alkaline Phosphatase↗
PubMed1997

Regional tissue blood flow and pH in the brain during deep hypothermic retrograde brain perfusion.

Deep hypothermic retrograde brain perfusion is used to protect the brain during aortic arch operations. However, all experiments have failed to demonstrate retrograde blood flow in the brain tissue. We developed an experimental model of sagittal sinus and simultaneous superior vena cava perfusion. Brain tissue blood flow was mapped with colored microspheres during deep hypothermic retrograde brain perfusion in 9 dogs. Regional brain pH was mapped photometrically using neutral red as a pH-indicating dye after 90 min of retrograde brain perfusion in 28 dogs and after 60 min of circulatory arrest in 8 dogs. Cerebral surface blood flow was also measured during retrograde brain perfusion. They were analyzed as functions of driving pressure between sagittal sinus and aorta. Total brain blood flow (ml/min/100 g) was 1.4 +/- 1.3, 3.8 +/- 2.6, and 4.6 +/- 2.6 when the driving pressure was 15, 25, and 35 mmHg, respectively (P < 0.05, 15 mmHg vs 25 mmHg). Regional cerebral blood flow (ml/min/100 g) with a driving pressure of 25 mmHg was 12.1 +/- 9.4, 7.0 +/- 5.6, 4.4 +/- 2.8, and 2.2 +/- 1.4 in the frontal cortex, anterior, mid, and posterior cerebrum, respectively. Cerebral cortex pH was 6.86 +/- 0.23, 7.15 +/- 0.18, and 6.46 +/- 0.13 after 90 min of retrograde brain perfusion with driving pressure of less than 20 mmHg, after that of above 20 mmHg, and after 60 min of circulatory arrest, respectively. Brain tissue pH, blood flows measured with microspheres, and laser flowmetry were highest when driving pressure was between 25 and 35 mmHg. We conclude that retrograde brain perfusion may provide maximum brain protection with driving pressure of 25 to 35 mmHg.

Animals↗
PubMed1997

Establishment and characterization of spontaneous mesothelioma cell lines derived from F344 rats.

Three mesothelioma cell lines (MeET-4, MeET-5, and MeET-6) established from ascitic fluid of F344 rats with spontaneous abdominal mesothelioma have been maintained through at least 60 passages on the DMEM with 10% FBS. Two of original tumours consisted of epithelioid cells growing in a papillary pattern, while one (original tumour of MeET-5) had sarcomatous areas composed of spindle-shaped tumour cells. The cell line originating from MeET-5 showed a constantly beiphasic growth pattern during the repetitive subcloning, while the other two lines retained a monophasic growth pattern. Although the growth pattern was different, the tumour cells in all three lines were positive for vimentin and keratin and ultrastructurally showed an abundant distribution of glycogen granules in the cytoplasm and numerous long microvilli on all surface. The modal chromosome number of cell lines varied from 41 to 71, and abnormal chromosomes were frequently seen. All cell lines established formed colonies on semi-solid medium and could be successfully transplanted, growing tumour masses in syngeneic rats and thus indicating their malignant nature. Cell lines grew even on a medium with a low concentration of FBS. The evidence suggests that they may produce growth factors that enable them to survive unfavourable medium conditions.

Abdominal Neoplasms↗
PubMed1997

Genetic polymorphism of the CYP2C subfamily and its effect on the pharmacokinetics of phenytoin in Japanese patients with epilepsy.

OBJECTIVE: To examine the genetic polymorphism of CYP2C9 and CYP2C19 and its effect on the pharmacokinetics of phenytoin among 44 Japanese patients with epilepsy. METHODS: Polymerase chain reaction tests with leukocyte deoxyribonucleic acid were used to detect the mutations for the amino acid substitution (Arg144-->Cys and Ile359-->Leu) in CgammaP2C9 and for the defective allele (m1 and m2) in CgammaP2C19. The pharmacokinetic parameters of phenytoin in individual patients were estimated by means of empirical bayesian analysis, in which the prior information was the population parameters for Japanese patients with epilepsy. RESULTS: Of the 44 patients, none had the CgammaP2C9 mutation for the Cys144 allele, whereas six patients were heterozygous for the wild-type (wt) and Leu359 allele (wt/Leu359) in cgammaP2C9. The maximal elimination rate (Vmax) of phenytoin among patients with heterozygous wt/Leu359 in CgammaP2C9 was 33% lower than that among patients with normal CgammaP2C9. A total of 21 patients were heterozygous for the CgammaP2C19 mutation (wt/m1 or wt/m2), and five patients had the homozygous or heterozygous mutations in CgammaP2C19 (m1/m1 or m1/m2). The Vmax values of phenytoin were slightly decreased (up to 14%) among patients with CgammaP2C19 mutations compared with patients with normal CgammaP2C19. CONCLUSION: The findings indicated that the genetic polymorphisms of CYP2C isozymes play an important role in the pharmacokinetic variability of phenytoin and that the mutation in CYP2C9 proteins (Ile359-->Leu) is a determinant of impaired metabolism of the drug among Japanese persons.

Adolescent↗
PubMed1997

Molecular form and subcellular distribution of acid beta-galactosidase in fibroblasts from patients with GM1 gangliosidosis, Morquio B disease and galactosialidosis.

The molecular form and subcellular distribution of acid beta-galactosidase in cultured fibroblasts from patients with beta-galactosidase deficiency (GM1-gangliosidosis, Morquio B disease and galactosialidosis) were studied, using antibodies against three different forms of the human enzyme: a high-molecular-weight multienzymic complex, a recombinant 84-kDa precursor, and a 64-kDa tryptic product of the precursor. The mature enzyme from normal fibroblasts was immunoprecipitated by the anti-complex and anti-64-kDa protein antibodies, but not by the anti-84-kDa precursor one. immunofluorescence staining of normal fibroblasts revealed the granular (lysosomal) distribution with anti-64-kDa protein antibody and the perinuclear reticular distribution with anti-84-kDa precursor antibody, probably representing the Golgi apparatus. Both patterns were demonstrated in Morquio B disease, but the residual enzyme activity was exclusively due to the mature enzyme. In Type 1 galactosialidosis, most of the expressed enzyme was detected as the precursor form with a perinuclear reticular distribution. In type 2 galactosialidosis, more than half of the enzyme activity was due to the mature form with a lysosomal distribution. Fibroblasts from a patient with GM1 gangliosidosis, expressing no beta-galactosidase mRNA, did not react against either anti-64-kDa protein antibody or anti-84-kDa precursor antibody. The combined use of immunoprecipitation and immunostaining was useful for analysing the pathophysiology of the intracellular processing and transport of the mutant beta-galactosidase.

Antibody Specificity↗
PubMed1997

Early-onset facioscapulohumeral muscular dystrophy: two case reports.

This report concerns two patients with facioscapulohumeral muscular dystrophy (FSHD) whose facial weakness began in infancy. In both patients, biopsied muscle histology showed mild myogenic changes accompanied by some regenerating and some small angular fibers, while endomysial inflammatory cellular infiltration was observed in Patient 1. The finding that our very young patients had muscle histopathological findings compatible with classical FSHD supports the previously expressed view that muscle histopathology is not related to either age or duration of the disease. Although Patient 2 was a sporadic case, both patients had the abnormal EcoRI DNA fragment detected by Southern blot analysis with probes p13E-11 and pFR-1, a finding compatible with FSHD. This indicates that gene analysis of sporadic cases must be as significant as that of familial cases. This report on patients with very early-onset and with common muscle histopathological and molecular genetic findings should contribute to widening the clinical spectrum of FSHD.

Adult↗
PubMed1997

Treatment of mitochondrial encephalomyopathy with a combination of cytochrome C and vitamins B1 and B2.

The therapeutic efficacy of a regimen consisting of intravenous injection of Cardiocrome, containing cytochrome c, flavin mononucleotide and thiamine diphosphate for mitochondrial encephalomyopathy (MEM) was examined. This combined therapy was applied to nine patients with MEM, including four with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes. For the standard regimen, Cardiocrome was first injected daily, usually for 4 weeks, and later by means of intermittent injections for maintenance treatment. Clinical improvement was obtained in eight of the patients. Improvement was observed in the muscle symptoms of easy fatigability, motor disability and severity of stroke-like episodes, as well as in various other symptoms such as phosphate, tinnitus, headache, corneal edema, chilblains, thalamic pain, respiratory failure, and nystagmus. This clinical improvement was maintained for more than 1 year by additional intermittent injections. In conclusion, this therapy was fairly effective for the management of patients with MEM.

Adolescent↗
PubMed1997

Evaluation of renal tubular secretion and reabsorption of levofloxacin in rats.

PURPOSE: Levofloxacin, a quinolone antibacterial drug, is a zwitterion at physiological pH. We examined the effect of cationic and anionic drugs on renal excretion of levofloxacin by means of in vivo clearance to characterize the mechanisms of renal excretion of this drug. METHODS: In vivo clearance was studied in male Wistar albino rats. A bolus dose of 2.85 mg/kg of levofloxacin was administered, followed by a constant infusion of 7.08 micrograms/min. Cimetidine, tetraethylammonium, or p-aminohippurate was administered as a bolus and incorporated into the infusion solution. After reaching steady state, urine and blood concentrations were measured, and pharmacokinetic parameters were calculated. RESULTS: Renal clearance was 2.56 +/- 0.42 ml/min in control, which accounted for 34% of the total body clearance. Renal clearance was significantly decreased to 0.83 +/- 0.25 ml/min by cimetidine (p < .05), corresponding to 32% of the control value. The cationic drug, tetraethyl-ammonium also reduced the renal clearance of levofloxacin, but the effect of the anionic drug, p-aminohippurate, was slight. The clearance ratio of levofloxacin, which was calculated by renal clearance divided by the plasma unbound fraction and the glomerular filtration rate, was 1.60 +/- 0.38 in the control and it was decreased to 0.68 +/- 0.17 and 1.11 +/- 0.22 by cimetidine and tetraethylammonium, respectively. CONCLUSIONS: The present results suggest that the renal excretion of levofloxacin in rats involves tubular secretion and reabsorption, in addition to glomerular filtration, and that tubular secretion is inhibited by cimetidine.

Animals↗
PubMed1997

Kinetic analysis of tetraethylammonium transport in the kidney epithelial cell line, LLC-PK1.

PURPOSE: The aims of this study were to establish a kinetic means of analyzing the membrane transport of organic cations in renal epithelial cells, and to simultaneously evaluate drug interactions in apical and basolateral membranes. METHODS: Tetraethylammonium (TEA) transport was measured using LLC-PK1 cell monolayers grown on microporous membrane filters. After incubating the cells with unlabeled TEA or other drugs, apical or basolateral medium was changed to that containing labeled TEA, and transcellular transport and cellular accumulation were measured. Clearance from apical medium to cells (CL12), cells to apical medium (CL21), cells to basolateral medium (CL23) and basolateral medium to cells (CL32) were calculated based on a three compartment model. RESULTS: TEA was accumulated progressively in the monolayers from the basolateral side and was transported unidirectionally to the apical side. CL32 was greater than CL12 and CL23 was greater than CL21. Therefore, the rate limiting step of TEA transport from the basolateral to the apical medium was the cell-to-apical step. Co-incubation of TEA with procainamide decreased the transport parameters of TEA, CL12, CL21 and CL32, whereas that with levofloxacin decreased only CL12 and CL21, not affecting the parameters in basolateral membranes. CONCLUSIONS: Using a simple model, we analyzed the transport of organic cation in kidney epithelial cell line, LLC-PK1. This method can be useful for the analysis of cation transport and drug interactions in the apical and basolateral membranes of renal tubules.

Animals↗
PubMed1997

Three-dimensional intraductal ultrasonography: preliminary results of a new technique for the diagnosis of diseases of the pancreatobiliary system.

BACKGROUND AND STUDY AIMS: Intraductal ultrasonography (IDUS) is a useful method for the diagnosis of pancreatobiliary diseases, but the images obtained using the presently available ultrasonic probes are limited by their two-dimensional nature and are sometimes difficult to interpret. The possible application of three-dimensional intraductal ultrasonography (3 D-IDUS) to facilitate the diagnosis of pancreatobiliary disease was therefore studied using a newly developed system. PATIENTS AND METHODS: 3 D-IDUS of the biliary system was performed on 26 patients with benign (n = 100) or malignant pancreatobiliary diseases (n = 16), via percutaneous transhepatic or peroral approaches. Three-dimensional diagnoses were made using both radial and linear images generated on the monitor of this system along with a comparison with conventional IDUS (2 D-IDUS) findings. RESULTS: The courses of vessels surrounding the bile duct could be easily imaged in all cases, and accurate assessment of tumor extension and the relationship with surrounding organs could be achieved for all the malignancies. In four of six cases of bile duct cancers the whole outline of the tumor could be visualized so that the tumor volumes could be measured. CONCLUSIONS: These preliminary results of 3 D-IDUS in the pancreatobiliary system, the first to be reported in the English literature, indicate that this new system has clear advantages over the 2 D-IDUS approach for diagnostic purposes. Further technical improvements can be expected which will ensure a clinical role for 3 D-IDUS.

Adult↗
PubMed1997

Unsuccessful omentopexy in thoracic surgery.

Omentopexy has improved the treatment of chronic empyema and postpneumonectomy bronchopleural fistula, which otherwise are difficult to manage. However, omentopexy is not effective in some patients. Four of 17 patients who underwent omentopexy in our institution between January 1978 and March 1994 did not respond to the treatment and died. In two patients, a dead space remained after surgery and there was insufficient sterilization. In one patient, a dead space appeared after surgery and it was impossible to control infection. The fourth patient had dehiscence of the anastomosis triggered by postoperative acute gastritis. All four patients in whom omentopexy was not successful died. When omentopexy is used for empyema, the space should first be sterilized. If sterilization is insufficient, muscle filling and thoracoplasty must be performed and no dead space left.

Adult↗
PubMed1997

"Reversed" latissimus dorsi musculocutaneous flap for closure of large bronchopleural fistula.

The patient was a 54-year-old male with diabetes mellitus and liver abscess perforating into the right lung through the diaphragm. After right lower lobectomy of the lung, S3-segmentectomy of the liver, and debridement of the subdiaphragmatic abscess a bronchopleural fistula appeared. After open-drainage thoractomy, secondary operation for closure of a large bronchopleural fistula and obliteration of the empyema cavities was performed with a "reversed" latissimus dorsi musculocutaneous flap.

Bronchial Fistula↗
PubMed1997

Haplotype and mutation analysis in Japanese patients with Wilson disease.

Wilson disease (WD), an autosomal recessive disorder of copper transport, is characterized by impaired biliary excretion and by impaired incorporation of copper into ceruloplasmin. Toxic accumulation of copper causes tissue damage, primarily in the liver, brain, and kidneys. The gene for WD (ATP7B) has been cloned, and the protein product is predicted to be a copper-transporting P-type ATPase with high amino acid identity with that for Menkes disease, an X-linked disorder of copper transport. Mutation screening in WD patients has led to the identification of at least 40 mutations. In addition, haplotype analysis using three dinucleotide-repeat markers, D13S314, D13S301, and D13S316, has been a useful indicator of specific mutations. We have determined haplotypes for the patients and their parents and sibs, in 21 unrelated WD families from Japan. Twenty-eight different haplotypes were observed on 42 WD chromosomes. In all the patients, the ATP7B coding sequence, including the intron-exon boundaries, was screened for mutations, by SSCP, followed by direct-sequence analysis of the shifted fragments. We identified 13 mutations, of which 11 mutations are novel, including 7 mutations-1 insertion, 4 deletions, and 2 missense mutations-in the coding region. The mutations reported in previous studies are 2299insC and Arg778Leu. Two patients were shown to have the 2299insC mutation, which has occurred in many different haplotypes in several populations, indicating a mutation hot spot. Primer-extension analysis of ATP7B mRNA has revealed multiple transcription start sites. Four of the novel mutations (three 1-bp changes and one 5-bp deletion) occur in the 5' UTR and may result in altered expression of the WD gene.

Adenosine Triphosphatases↗
PubMed1997

Renal excretion of vancomycin in rats with acute renal failure.

We have investigated the renal excretion of vancomycin in rats with acute renal failure (ARF) induced by uranyl nitrate or cisplatin. The renal clearance of the antibiotic after uranyl nitrate or cisplatin injection was separately evaluated by calculating the glomerular filtration rate (GFR) and secretory clearance. The reduced renal clearance of vancomycin in these ARF rats was a result of a decrease in both GFR and secretory clearance. The extents of the decreases in GFR and in secretory clearance were not, however, proportional, the extent of the decrease in secretory clearance being more pronounced. These results suggest that the renal tubular secretion of vancomycin was reduced more predominantly than glomerular filtration in these ARF models.

Acute Kidney Injury↗
PubMed1997

Inhibition of PAH transport by parathyroid hormone in OK cells: involvement of protein kinase C pathway.

We have previously shown that the p-aminohippurate (PAH) transport system in OK kidney epithelial cell line is under the regulatory control of protein kinase C. Parathyroid hormone (PTH) could activate protein kinase C, as well as protein kinase A, in OK cells. In the present study, the effect of PTH on PAH transport was studied in OK cells. PTH inhibited the transcellular transport of PAH from the basal to the apical side, as well as the accumulation of PAH in OK cells. Basolateral PAH uptake was inhibited by PTH in a dose- and time-dependent manner. Protein kinase A activators did not affect the transcellular transport or the accumulation of PAH. The PTH-induced inhibition of the accumulation of PAH was blocked by a protein kinase C inhibitor staurosporine. These results suggest that PTH inhibits the PAH transport in OK cells and that the messenger system mediated by protein kinase C, not protein kinase A, plays an important role in the regulation of PAH transport by PTH.

1-Methyl-3-isobutylxanthine↗
PubMed1997

Recognition of beta-lactam antibiotics by rat peptide transporters, PEPT1 and PEPT2, in LLC-PK1 cells.

PEPT1 and PEPT2 are H(+)-coupled peptide transporters expressed preferentially in the intestine and kidney, respectively, which mediate uphill transport of oligopeptides and peptide-like drugs such as beta-lactam antibiotics. In the present study, we have compared the recognition of beta-lactam antibiotics by LLC-PK1 cells stably transfected with PEPT1 or PEPT2 cDNA. Cyclacillin (aminopenicillin) and ceftibuten (anionic cephalosporin without an alpha-amino group) showed potent inhibitory effects on the glycylsarcosine uptake in the PEPT1-expressing cells. Other beta-lactams, such as cephalexin, cefadroxil, and cephradine (aminocephalosporins), inhibited modestly the PEPT1-mediated glycylsarcosine uptake. Except for ceftibuten, these beta-lactams showed much more potent inhibitions on the glycylsarcosine uptake via PEPT2 than via PEPT1. Comparison of the inhibition constant (Ki) values between cefadroxil and cephalexin suggested that the hydroxyl group at the NH2-terminal phenyl ring increased affinity for both PEPT1 and PEPT2. It is concluded that PEPT2 has a much higher affinity for beta-lactam antibiotics having an alpha-amino group than PEPT1 and that substituents at the NH2-terminal side chain of these drugs are involved in the recognition by both peptide transporters.

Animals↗
PubMed1997

[The studies of extracorporeal shock wave lithotripsy (ESWL) for pancreatic ductal stones].

30 patients with main pancreatic duct stones were treated by ESWL. In 18 of 22 patients who had not previously undergone endoscopic pancreatic sphincterotomy (EPST) or endoscopic sphincterotomy (EST), the stone fragments disappeared after ESWL. The fragments were removed endoscopically in the remaining 4 cases. Complete clearance was achieved in 8 cases with endoscopically unextractable stones by ESWL. After the ESWL procedure, absolute relief from pain was reported by in 19 of 22 patients with abdominal complaints. Serum amylase levels decreased significantly, and dilatation of the main pancreatic duct (MPD) was reduced. In the medium-term follow-up period, pancreatic exocrine function and endocrine function had a possibility to be preserved. One case of pancreatic cancer and one case of an intraductal papillary tumor of the pancreas were found, indicating that careful observation is necessary even after complete removal of pancreatic stones. In cases of Santorini duct dominant, multiple stones, or stricture of the MPD, ESWL should be combined with EPST and endoscopic stenting for preventing recurrence of acute pancreatitis and pancreatic stones. In conclusion, ESWL is the first choice of treatment for pancreatolithiasis and useful procedure and the limited complications.

Adult↗