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M Kitada

Publications and source records attributed to M Kitada.

At least 55 records · Page 3Linked to original sources
PubMed1999

Enantioselectivity of bunitrolol 4-hydroxylation is reversed by the change of an amino acid residue from valine to methionine at position 374 of cytochrome P450-2D6.

The enantioselectivity of 4-hydroxylation of bunitrolol (BTL), a beta-adrenoceptor blocking drug, was studied in microsomes from human liver, human hepatoma (Hep G2) cells expressing CYP2D6, and lymphoblastoid cells expressing CYP2D6. Kinetics in human liver microsomes showed that the Vmax value for (+)-BTL was 2.1-fold that of (-)-BTL, and that the Km value for (+)-BTL was lower than that for the (-)-antipode, resulting in the intrinsic clearance (Vmax/Km) of (+)-BTL being 2.1-fold over its (-)-antipode. CYP2D6 (CYP2D6-met) expressed in Hep G2 cells had a methionine residue at position 373 of the amino acid sequence and a rat-type N-terminal peptide (MELLNGTGLWSM) instead of the human-type (MGLEALVPLAVIV), and showed enantioselectivity of [(+)-BTL < (-)-BTL] for the rate of BTL 4-hydroxylation. In contrast, enantioselectivity [(+)-BTL > (-)-BTL] for Hep G2-CYP2D6 (CYP2D6-val) with a human-type N-terminal peptide that had a valine residue at 374, which corresponds to the methionine of the CYP2D6-met variant, was the same as that for human liver microsomes. We further confirmed that CYP2D6-met and CYP2D6-val expressed in human lymphoblastoid cells, both of which have methionine and valine, respectively, at position 374 and a human-type N-terminal peptide, exhibited the same enantioselectivities as those obtained from CYP2D6-met and CYP2D6-val expressed in the Hep G2 cell system. These results indicate that the amino acid at 374 of CYP2D6 is one of the key factors influencing the enantioselectivity of BTL 4-hydroxylation.

Adrenergic beta-Antagonists↗
PubMed1999

Distribution of synaptosomal-associated protein 25 in nerve growth cones and reduction of neurite outgrowth by botulinum neurotoxin A without altering growth cone morphology in dorsal root ganglion neurons and PC-12 cells.

Synaptosomal-associated protein 25 has been regarded as one of the target-associated soluble N-ethylmaleimide-sensitive fusion attachment protein receptors essential for exocytosis of vesicles in synapses. We have previously reported that cleavage of syntaxin, which is another target-associated soluble N-ethylmaleimide-sensitive fusion attachment protein receptor, with botulinum neurotoxin C1 resulted in inhibition of neurite extension and morphological changes including growth cone collapse and large vacuole formation. As an attempt to explore the mechanism of growth cone extension, we examined the ultrastructural localization of synaptosomal-associated protein 25 in growth cones with or without treatment of botulinum neurotoxin A, which cleaves synaptosomal-associated protein 25. In dorsal root ganglion neurons, light microscopy demonstrated synaptosomal-associated protein 25 immunoreactivity throughout the neurons, including the cell bodies, neurites and growth cones. Using electron microscopy, gold signals immunoreactive for synaptosomal-associated protein 25 were identified diffusely in the cytoplasm of the growth cones. In contrast, in PC-12 cells, a large number of gold signals were localized on the plasma membranes. High levels of signal were also found in the cytoplasm in the central region of the growth cones. We also confirmed that botulinum neurotoxin A treatment reduced neurite extension by about 50%. However, both in dorsal root ganglion neurons and in PC-12 cells we found no differences in the ultrastructure nor in the localization of synaptosomal-associated protein 25 between growth cones with and without toxin treatment. These results indicate that cleavage of synaptosomal-associated protein 25 inhibits growth cone extension in a manner different than that of syntaxin cleavage. The results of this study suggest the possibility that synaptosomal-associated protein 25 is involved in growth cone extension through a process independent of vesicle fusion.

Animals↗
PubMed1999

Comparison of urinary 6beta-hydroxycortisol/cortisol ratio between neonates and their mothers.

AIMS: To assess CYP3A enzyme activity in human neonates by measuring the urinary 6beta-hydroxycortisol/cortisol (6beta-OHF/C) ratio. METHODS: Fifty-six mature male neonates with normal delivery, seventeen of their mothers and twenty-four healthy non-pregnant young women participated in this study. Urinary 6beta-OHF/C ratio was determined on the day of birth in neonates and their mothers. In addition, changes in the ratio after birth were determined in neonates. RESULTS: On the day of birth, the urinary 6beta-OHF/C ratio of neonates was significantly higher than that of their mothers (20.5 vs 6.9). In contrast, no significant difference was observed in the mean ratio of urinary 6beta-OHF/C between women with and without pregnancy (6.9 vs 9.0). The urinary 6beta-OHF/C ratio after birth was decreased day by day in neonates. CONCLUSION: These results indicate that the high urinary 6beta-OHF/C ratio in mature neonates on the day of birth is independent of the activity of CYP3A enzyme in their mothers.

Aryl Hydrocarbon Hydroxylases↗
PubMed1999

Estimation of chemical structure of notopterol metabolites.

Notopterol, 5[(2E)-5-hydroxy-3,7-dimethyl-2,6-octadienyloxy]psoralen showed an inhibitory effect on aminopyrine N-demethylase activity in liver microsomes. In addition, notopterol has been found to be metabolized by cytochrome P450, and two kinds of metabolites were formed from notopterol. Furthermore, specific cytochrome P450 3A4 inhibitors which were isolated from grapefruit juice had the same furocoumarin structure as notopterol. Two metabolites of notopterol were separated by HPLC, and the chemical structures of the hydroxylated metabolites were estimated by 1H-NMR spectra and liquid chromatography-mass spectrometry.

Animals↗
PubMed1999

[A case of tracheal fistula after operation for esophageal cancer].

We have experienced a case of mediastinal abscess and tracheal fistula after operation for esophageal cancer and successfully closed by using intercostal muscle pedicle flap. A 61-year-old male underwent esophagectomy for advanced esophageal cancer. On the 12th postoperative day, mediastinal abscess caused by leakage was detected, and drainage of the mediastinal and thoracic cavity was performed. On the 29th postoperative day, tracheal fistula was detected, and operation was performed in order to close the fistula by using of intercostal muscle pedicle flap. His postoperative course was fair and general condition was improvement, esophageal reconstruction using of free jejunal graft was performed and oral ingestion was started.

Aged↗
PubMed1999

[Microwave coagulation therapy under laparotomic ischemia for multiple liver metastases of colorectal cancer].

Microwave coagulation therapy (MCT) under laparotomic ischemia induced by partial obstruction of the hepatic artery and portal vein was conducted on patients with multiple liver metastases of colorectal cancer. The patients were then compared with those who underwent non-ischemic MCT. Among the patients with liver metastasis of colorectal cancer we encountered between August 1990 and October 1998, 14 patients who developed multiple cancer (five or more) in the bilateral liver lobes were enrolled in the study. No clear differences were observed in the sex, age, frequency of simultaneousness, therapy other than MCT, number of foci, and number of MCT between the ischemic MCT and non-ischemic MCT group. Postoperative CT revealed residual foci in one of the seven patients in the ischemic MCT group. A comparison of the cumulative survival rate revealed that the ischemic MCT group had a higher one-year survival rate (50%) than the non-ischemic MCT group (14%). A comparison of patients with a residual lesion and those with no residual lesion showed that all six patients with a residual lesion died less than one year after surgery. Eight patients with no residual lesion had a significantly better prognosis (p < 0.05). It is important to eliminate any residual metastatic lesion during surgery in multiple liver metastases of colorectal cancer If the residual lesion is non-resectable, its elimination by ischemic MCT would contribute to the long-term survival of the patients.

Adult↗
PubMed1998

Differential catalytic properties in metabolism of endogenous and exogenous substrates among CYP3A enzymes expressed in COS-7 cells.

The catalytic properties of CYP3A7 in the metabolism of endogenous and exogenous substrates were compared with those of CYP3A4 and CYP3A5 using COS-7 expressing enzymes. The highest activities of dehydroepiandrosterone (DHEA) and dehydroepiandrosterone 3-sulfate (DHEA-S) 16alpha-hydroxylase were observed in COS-7 cells expressing CYP3A7. In contrast, the activity of testosterone 6beta-hydroxylase of CYP3A7 expressed in COS-7 cells was much less than that of CYP3A4 expressed in COS-7 cells. The rate of carbamazepine 10, 11-epoxidation was the greatest in COS-7 cells expressing CYP3A4, followed by CYP3A5 and CYP3A7. On the other hand, the formation of reductive metabolite of zonisamide was the highest in COS-7 cells expressing CYP3A4, followed by CYP3A7 and CYP3A5. Furthermore, the addition of triazolam resulted in a decrease in 6beta-hydroxylation catalyzed by CYP3A7, but not by CYP3A4, whereas the pretreatment of microsomes with triacetyloleandomycin (TAO) resulted in a decrease in the reaction catalyzed by CYP3A4, but not by CYP3A7. Together with these results, it was suggested that CYP3A7 exerts differential catalytic properties not only in metabolism of endogenous substrates but also in drug metabolism compared to CYP3A4 and CYP3A5.

Adult↗
PubMed1998

Long acellular nerve transplants for allogeneic grafting and the effects of basic fibroblast growth factor on the growth of regenerating axons in dogs: a preliminary report.

Sciatic nerves were excised from 3 beagle dogs about 5 h after their sacrifice, treated three times by freezing and thawing, and stored in physiological saline for 3 months at -20 degrees C until used. Nerve segments 5 cm in length prepared from these stored nerves were transplanted to the common peroneal nerve in the right hindlimb of beagle dogs. Sixteen beagle dogs in total were used, in four treatment groups of two pairs each studied at 1 and 3 months. Five-hundred microliters basic fibroblast growth factor (bFGF) of two different concentrations (10 micrograms/300 microliters and 100 micrograms/300 microliters) which were impregnated in 0.5 ml gelatin hydrogels was applied around the sutured allografts. Autografting was also done in 4 beagle dogs, with no bFGF application. One month after the grafting, no regenerating nerves extended beyond the middle of the transplant in any of the allografts, except in the autografts in which a number of regenerated (myelinated) axons were present. Three months after the grafting, an abundance of myelinated axons was found at the middle of the graft: the numbers of axons per 10(4) micron 2 were 22.6 in the autografts and 10.6, 10.4 and 19.2 in the allografts treated with no bFGF, low-dose bFGF, and high-dose bFGF, respectively. Regenerating axons extended into the host nerve: the numbers of myelinated axons at the level 1.5 cm distal to the distal suture were 35.7, 0.9, 3.8, and 12.1 per 10(4) micron 2 in the above respective order. Although it was inferior in quality to the autograft, peripheral nerve regeneration was extensive in the distal nerve using freeze-thawed and bFGF-treated allografts at 3 months. Electromyography showed that the peroneus longus muscle responded to the electrical stimuli given at the site proximal to the transplant in all four groups. These data indicate that a 5-cm acellular nerve segment containing Schwann cell basal laminae can be used successfully as an allograft without any immunosuppressants and that exogenously applied bFGF can improve nerve regeneration by enhancing the growth of regenerating axons.

Animals↗
PubMed1998

Changes in urinary 6beta-hydroxycortisol/cortisol ratio after birth in human neonates.

OBJECTIVE: Urinary 6beta-hydroxycortisol/cortisol (6beta-OHF/C) ratio was measured in human neonates to assess the CYP3A enzyme activity. METHODS: Urinary 6beta-OHF/C ratio was determined on the day of birth in 94 neonates including those born prematurely. In addition, changes in the ratios after birth were also determined in 81 neonates. RESULTS: On the day of birth, a significant positive correlation was found between urinary 6beta-OHF/C ratios and gestational age (r = 0.476) and birth weight (r = 0.283). There was no gender difference in the urinary 6beta-OHF/C ratios in human neonates. Furthermore, delivery modes such as cesarean section and vaginal delivery did not appear to affect the urinary 6beta-OHF/C ratio. The mean ratio of urinary 6beta-OHF/C observed in 39 mature neonates (more than 37 weeks of gestational age) was higher than that observed in adults (16.5 vs 9.9). Within 5 days after birth, the ratio rapidly decreased to less than that in adults. In contrast, the mean ratio of urinary 6beta-OHF/C observed in 42 premature neonates (under 37 weeks of gestational age) was significantly lower than that observed in mature neonates (5.3 vs 16.5) and was virtually unchanged during the 14-days after birth. Therefore, no significant difference was observed in the mean ratio of urinary 6beta-OHF/C between mature and premature neonates at 5 days after birth. CONCLUSION: From these results, it was concluded that on the day of birth, mature neonates might possess a higher activity of CYP3A enzyme compared with premature neonates, and that the CYP3A enzyme activity in mature neonates might be promptly changed at an early stage after birth.

Aryl Hydrocarbon Hydroxylases↗
PubMed1998

Prediction of drug-drug interactions of zonisamide metabolism in humans from in vitro data.

OBJECTIVE: The purposes of this study were to identify the P450 enzyme (CYP) responsible for zonisamide metabolism in humans by using expressed human CYPs and to predict drug interaction of zonisamide in vivo from in vitro data. METHODS: Ten expressed human CYPs and human liver microsomes were used in the experiments for the identification of enzymes responsible for zonisamide metabolism and for the prediction of drug-drug interactions of zonisamide metabolism in humans from in vitro data, respectively. Two-sulfamoylacetyl phenol, a reductive metabolite of zonisamide, was measured by the HPLC method. RESULTS: From the experiments using ten expressed human CYPs, CYP2C19, CYP3A4 and CYP3A5 were shown to be capable of catalyzing zonisamide reduction. However, an intrinsic clearance, Vmax/kM, of CYP3A4 was much higher than those of CYP2C19 and CYP3A5. From the point of view of enzyme amount in human liver CYPs isoform and their intrinsic clearance, it was suggested that CYP3A4 is mainly responsible for zonisamide metabolism in human CYPs. Zonisamide metabolism in human liver microsomes was markedly inhibited by cyclosporin A, dihydroergotamine, ketoconazole, itraconazole, miconazole and triazolam. We estimated the possibility and degree of change of zonisamide clearance in vivo in clinical dose range from in vitro inhibition constant of other drugs against zonisamide metabolism (Ki) and unbound inhibitor concentration in blood (Iu) in clinical usage. Clearance of zonisamide was maximally estimated to decrease by 31%, 23% and 17% of the clearance without inhibitors i.e. ketoconazole, cyclospolin A and miconazole, respectively. Fluconazole and carbamazepine are estimated to decrease by 5-6% of the clearance of zonisamide. On the other hand, there may be lack of interaction of zonisamide metabolism by dihydroergotamine, itraconazole and triazolam in clinical dose range. CONCLUSION: We demonstrated that: (1) zonisamide is metabolized by recombinant CYP3A4, CYP2C19 and CYP3A5, (2) the metabolism is inhibited to a variable extent by known CYP3A4/5 substrates and/or inhibitors in human liver microsomes, and (3) in vitro-in vivo predictive calculations suggest that several compounds demonstrating CYP3A4-affinity might cause in vivo drug-drug interactions with zonisamide.

Anticonvulsants↗
PubMed1998

Trimethadione metabolism and microsomal monooxygenases in untreated and phenobarbital-treated rhesus monkeys.

The contribution of induced cytochrome P450 (P450) isozymes (CMLa; CYP2B, CMLb; CYP2A and CMLc; CYP3A) and related enzymes to trimethadione (TMO) metabolism in phenobarbital-treated rhesus monkey were investigated. The animals received a single dose of TMO (4 mg kg-1) and plasma samples were withdrawn before this administration and again at 0.08, 0.25, 0.5, 1 and 2 h later. Phenobarbital-treatment (20 mg kg-1 day-1 for 3 days; i.p.) significantly increased the plasma dimethadione (DMO)/TMO ratios at 0.08, 0.5, 1 and 2 h one's appropriate controls. Phenobarbital treatment also increased the P450 content (1.7-fold) and activity of aniline p-hydroxylase (1.3-fold), p-nitroanisole O-demethylase (1.8-fold) and benzphetamine N-demethylase (2.3-fold). The content of CMLa, CMLb and CMLc were increased about 12.8, 2.3 and 2.7-fold by phenobarbital pretreatment, respectively. The activity of TMO N-demethylation was inhibited by anti-P450 CMLa and anti-P450 CMLb. However, the anti-P450 CMLc antibody had no effect on this activity in liver microsomes. The results of both in vivo and in vitro studies of the effects of phenobarbital treatment on TMO metabolism indicate that these effects may be attributed to the induction of CMLa. These findings suggest that plasma DMO/TMO ratio in a single blood sampling after TMO administration is very useful for determination the degree of hepatic induction in clinical study.

Aniline Hydroxylase↗
PubMed1998

Nucleotide and amino acid sequences of the monkey P450 2B gene subfamily.

The nucleotide sequence of a cDNA coding for monkey cytochrome P450 (P450) 2B has been determined. Using antibody against P450 CMLa which had been purified from hepatic microsomes of untreated cynomolgus monkeys, a cDNA clone with 2,275 bp insert (Mac2B) was isolated from a gamma gt11 cDNA library prepared from hepatic mRNA from an untreated rhesus monkey. The cloned insert was sequenced and found to contain an open reading frame coding for a polypeptide of 491 amino acids. The molecular weight calculated from the deduced amino acid sequence was 55,969. The N-terminal 34 amino acids encoded by Mac2B were identical to those determined by Edman degradation analysis of purified cynomolgus monkey P450 CMLa. The nucleotide and the deduced amino acid sequences of Mac2B which is now called CYP2B17 were the most similar to those of human P450 2B6 among the P450 2B subfamily and those sequences of Mac2B were 94% and 90% identical to those of human P450 2B6, respectively.

Amino Acid Sequence↗
PubMed1998

Metabolism of dacarbazine by rat liver microsomes contribution of CYP1A enzymes to dacarbazine N-demethylation.

The N-demethylation of dacarbazine in liver microsomes was significantly increased by treatment of rats with beta-naphthoflavone, dexamethasone, or phenobarbital. However, the extent of increase in the N-demethylation observed in beta-naphthoflavone-treated rats was much greater than that observed in dexamethasone-treated rats. A good correlation between N-demethylation of dacarbazine and O-deethylation of phenacetin was observed when a low concentration of phenacetin was used. Furthermore, the activity of dacarbazine N-demethylase in rat liver microsomes was highly correlated with the amounts of CYP protein immunochemically determined with anti-rat CYP1A2 antibodies. In addition, antibodies to rat CYP1A2, and furafylline and alpha-naphthoflavone, which are known inhibitors of CYP1A enzymes, exhibited inhibitory effects on dacarbazine N-demethylation. These results indicated that CYP1A enzymes may be responsible for N-demethylation of dacarbazine in rat liver microsomes.

Animals↗
PubMed1998

Steroid hydroxylation by human fetal CYP3A7 and human NADPH-cytochrome P450 reductase coexpressed in insect cells using baculovirus.

Human fetal CYP3A7 and human NADPH-cytochrome P450 reductase were coexpressed in insect cells, TN-5, infected with a recombinant baculovirus carrying both cDNAs. The expression of reductase in TN-5 cells was shown to be sufficient for the CYP3A7 dependent 16 alpha-hydroxylation of dehydroepiandrosterone. However, the extra addition of cytochrome b5 and phospholipid was necessary to obtain a maximal activity of CYP3A7 catalyzing the reaction. CYP3A7 expressed in TN-5 cells was capable of metabolizing testosterone, cortisol and dehydroepiandrosterone 3-sulfate as well as dehydroepiandrosterone. The apparent Vmax for 6 beta-hydroxylations of testosterone was similar to that obtained for 6 beta-hydroxylation of cortisol (2.9 versus 2.5 nmol/nmolP450/min). In contrast, the apparent Vmax for 16 alpha-hydroxylation of dehydroepiandrosterone and its 3-sulfate were 20 and 2 times greater than those observed for steroid 6 beta-hydroxylations, respectively (67.5 and 5.8 versus 2.5-2.9 nmol/nmol P450/min). On the other hand, the apparent K(m) for 6 beta-hydroxylations of testosterone and cortisol were greater than those for 16 alpha-hydroxylations (120 and 860 versus 46-58 microM). Thus, CYP3A7 was active for steroid 6 beta-hydroxylations and 16 alpha-hydroxylations, but there were greater differences in Vmax/K(m) ratios between these reactions.

Animals↗
PubMed1998

[Good response in case of hepatocellular carcinoma; a case report of interdisciplinary local therapy].

A 47-year-old male patient with chronic hepatitis had a high AFP level during the follow-up period. Abdominal CT revealed at S5, which led to a diagnosis of hepatocellular carcinoma. In August 1992, partial resection of S5 was performed, and ethanol was injected into the tumor at S2. In July 1993, recurrent tumors were observed at S5 and S3. PEIT was performed for each lesion. In December 1994, multiple recurrence was observed and 4 mg SMANCS was injected through the proper hepatic artery. In July 1995, another 4 mg of SMANCS was injected into the tumors. In June 1996, only a 20 mm lesion at S5 remained while the other lesions disappeared. Under general anesthesia, the patient underwent percutaneous microwave tumor coagulation. In December 1997, the AFP level was normal, and imaging revealed disappearance of the recurrent tumor. Selection of local therapy for hepatocellular carcinoma achieved long survival in our case considering the QOL and frequent therapy administration.

Antineoplastic Agents↗
PubMed1998

Stimulation of cholesterol release from rabbit foam cells by the action of a new inhibitor for acyl CoA:cholesterol acyltransferase (ACAT), HL-004.

A new inhibitor of acyl CoA:cholesterol acyltransferase (ACAT), HL-004 [N-(2, 6-diisopropylphenyl)tetradecylthioacetamide], suppressed the synthesis of cholesterol [14C]oleate at 10(-9) approximately 10(-7) M in a concentration-dependent manner in both THP-1 cell-derived macrophages and foam cells prepared from aortic intima of rabbits fed a high cholesterol diet. Incorporation of [3H]cholesterol oleate-beta very low density lipoproteins was not inhibited by HL-004 at 10(-9) approximately 10(-7) M. Release of radioactivity from the cells loaded with [3H]cholesterol oleate-beta very low density lipoproteins was increased by the inhibition of ACAT activity with HL-004. HL-004 did not affect on acid and neutral cholesterol esterases. As a result, cholesterol ester content in foam cells decreased. These data suggested that HL-004 decreases cholesterol ester in foam cells by increasing the release of cholesterol and therefore might suppress atherosclerotic lesions.

Acetanilides↗
PubMed1997

K+/H+ antiporter in alkaliphilic Bacillus sp. no. 66 (JCM 9763).

A K+/H+ antiport system was detected for the first time in right-side-out membrane vesicles prepared from alkaliphilic Bacillus sp. no. 66 (JCM 9763). An outwardly directed K+ gradient (intravesicular K+ concentration, K(in), 100 mM; extravesicular K+ concentration, K(out), 0.25 mM) stimulated uphill H+ influx into right-side-out vesicles and created the inside-acidic pH gradient (delta pH). This H+ influx was pH-dependent and increased as the pH increased from 6.8 to 8.4. Addition of 100 microM quinine inhibited the H+ influx by 75%. This exchange process was electroneutral, and the H+ influx was not stimulated by the imposition of the membrane potential (interior negative). Addition of K+ at the point of maximum delta pH caused a rapid K+-dependent H+ efflux consistent with the inward exchange of external K+ for internal H+ by a K+/H+ antiporter. Rb+ and Cs+ could replace K+ but Na+ and Li+ could not. The H+ efflux rate was a hyperbolic function of K+ and increased with increasing extravesicular pH (pH(out)) from 7.5 to 8.5. These findings were consistent with the presence of K+/H+ antiport activity in these membrane vesicles.

Antiporters↗