Cloning and expression analysis of a new member of the cytochrome P450, CYP2A15 from the Chinese hamster, encoding testosterone 7alpha-hydroxylase.

We cloned a new cytochrome P450 cDNA encoding testosterone 7alpha-hydroxylase in the Chinese hamster, designated CYP2A15 which shares significant amino acid sequence homology with members of the CYP2A subfamily. The CYP2A15 cDNA was isolated by screening a liver cDNA library and the sequence contains an open reading frame of 1482 nucleotides encoding a polypeptide of 493 amino acids with a calculated molecular mass of 56,295 Da. This is flanked by a 5'-untranslated region of 2 bp and a 3' untranslated region of 191 bp including the poly(A) tail.

Molecular cloning, heterologous expression, and characterization of a novel member of CYP2A in the Syrian hamster.

The cDNA clone coding for a novel cytochrome P-450 2A subfamily member (CYP2A16) was isolated from a Syrian hamster liver cDNA library. The deduced amino acid sequence of CYP2A16 showed more than 90% identity with those of rat CYP2A3 and mouse CYP2A4/5. The catalytic activity of CYP2A16 was determined by transient expression of its cDNA in transfected COS7 cells and CYP2A16 was found to have the testosterone 2 beta-, 15 alpha-, and 15 beta-hydroxylases, coumarin 7-hydroxylase, and ethoxycoumarin O-deethylase activities.

Polyunsaturated fatty acid anilides as inhibitors of acyl-coA: cholesterol acyltransferase (ACAT).

A series of polyunsaturated fatty acid anilides were synthesized and evaluated as ACAT inhibitors. Compound 24 had potent inhibitory activity against microsomal ACAT derived from U937, HepG2 and Caco-2 cell lines. Therefore, it might be expected to act as an antiarteriosclerotic and hypocholesterolemic agent.

Effects of interferon on circadian changes in blood pressure and heart rate variability in patients with chronic hepatitis.

We have determined the effects of interferon therapy on circadian changes in blood pressure and heart rate variability in normotensive hospitalized patients with chronic active hepatitis. Body temperature and pulse rate increased for the initial few days of interferon therapy without significant change in casual or ambulatory blood pressure. Interferon therapy failed to elicit any significant changes in the power spectrum of R-R intervals.

Differential response of angiotensin peptides in the urine of hypertensive animals.

Urinary excretion rates of angiotensin I (Ang I), angiotensin II (Ang II), and angiotensin-(1-7) [Ang-(1-7)] were determined in normotensive Sprague Dawley (SD), spontaneously hypertensive (SHR), and mRen-2 transgenic hypertensive animals before and following blockade of Ang II synthesis or activity for two weeks. This study was performed to determine for the first time whether inhibition of Ang II alters the excretion of angiotensin peptides in the urine. Rats were given either tap water or water medicated with lisinopril, losartan or both agents in combination.

Estimation of absorption of trihalomethanes and carbon tetrachloride in low-level exposure by inhalation pharmacokinetic analysis in rats.

Inhalation pharmacokinetics of the environmental contaminants chloroform, bromodichloromethane, chlorodibromomethane, and carbon tetrachloride were evaluated, and their absorption amounts were estimated in rats to obtain fundamental data for risk assessment of chronic low-level exposures. Measured amounts of the substances were injected into a closed chamber system in which a rat had been placed, and the concentration changes in the chamber were examined. The pharmacokinetics of the substances were evaluated on the basis of the concentration-time courses using linear or nonlinear compartment models.

A novel positive regulatory element that enhances hamster CYP2A8 gene expression mediated by xenobiotic responsive element.

CYP2A8 is a major form of cytochrome P-450 inducible by 3-methylcholanthrene in Syrian hamster liver. To identify DNA elements necessary for the transcriptional activation of the CYP2A8 gene, we analyzed the regulatory region of the CYP2A8 gene and conducted transient transfection experiments of CYP2A8-luciferase fusion plasmids in primary cultures of hamster hepatocytes. We analyzed up to -5 kb of the 5'-flanking region and found the region sufficient for the 3-methylcholanthrene-inducible gene expression.

A major phenobarbital-inducible P450 isozyme, CYP2A14, in the Chinese hamster liver: purification, characterization, and cDNA cloning.

Phenobarbital, a potent inducer of CYP2B isozyme of cytochrome P450, induces mainly CYP2A but not CYP2B in the Chinese hamster liver. A major isozyme inducible by phenobarbital was purified by column chromatography from Chinese hamster livers. This isozyme, named P450CH2A-2 and designated CYP2A14, had in the reconstituted system high activities of 7-ethoxycoumarin O-deethylase (43 nmol/min/nmol P450) and aflatoxin B1 activation and a moderate activity of testosterone 15alpha-hydroxylase and coumarin 7-hydroxylase.

Regression of carotid wall thickening after corticosteroid therapy in Takayasu's arteritis evaluated by B-mode ultrasonography: report of 2 cases.

We describe 2 young Japanese female patients with active Takayasu's arteritis whose diagnosis was mainly based on the characteristic ultrasonographic finding, that is, a circumferential vascular wall thickening in common carotid arteries. Following corticosteroid therapy, serial ultrasonographic examination documented the improvement of their thickened carotid walls (in Case 1 from 1.4 to 0.

Evidence for involvement of cAMP-dependent pathway in the phenobarbital-induced expression of a novel hamster cytochrome P450, CYP3A31.

Recently, we isolated a novel Syrian hamster cDNA clone that encodes a protein which has been named CYP3A31. In primary hepatocyte cultures, CYP3A31 is dramatically induced by phenobarbital. To elucidate the mechanism of this induction, we first studied the effects of cAMP on phenobarbital-induced CYP3A31 expression using forskolin and N6,O2'-dibutyryl cAMP in hepatocyte cultures.

Modulation of snake hepatic cytochrome P450 by 3-methylcholanthrene and phenobarbital.

Induction mode of the hepatic drug-metabolizing enzymes was studied in Corn snake (Elaphe guttata emoryi). Treatment of snakes with 3-methylcholanthrene or phenobarbital produced no effects on liver weight and total content of cytochromes P450 and b5. Treatment with 3-methylcholanthrene significantly induced the activities of arylhydrocarbon hydroxylase, 7-ethoxyresorufin O-deethylase and 7-pentoxyresorufin O-dealkylase, whereas those of ethoxycoumarin O-deethylase, benzphetamine N-demethylase, erythromycin N-demethylase and testosterone hydroxylases were not affected.

Interaction of opioids and vasopressin in central action of angiotensin II in conscious rabbits.

It has been demonstrated that opioids modulate the renin-angiotensin and sympathetic nervous system. To clarify the interaction of central angiotensin II (Ang II) and endogenous opioids, we studied the effects of naloxone, an opioid antagonist, on cardiovascular and sympathetic responses to intracerebroventricular (i.c.

Estimation of absorption of environmental contaminants in low-level exposure by pharmacokinetic analysis.

Inhalation pharmacokinetics of the environmental contaminants 1,1,1-trichloroethane, tetrachloroethylene, trichloroethylene, benzene, and p-dichlorobenzene were determined in male Sprague-Dawley rats, and data generated were used to obtain fundamental data for risk assessment of chronic low-level exposures to these substances. Measured amounts of the substances were injected into a closed chamber system in which a rat had been placed, and the concentration changes in the chamber air were examined. The pharmacokinetics of the substances were evaluated using linear or nonlinear compartment models.

Positive and negative affect in the factor structure of the State-Trait Anxiety Inventory for Japanese workers.

The factor structure of Form Y of the State-Trait Anxiety Inventory (STAI-Y) was examined with 1,862 Japanese adult workers (1,509 men, 353 women). The initial principal component analysis extracted three factors based on the scree test. All 20 state (S-Anxiety) and 20 trait (T-Anxiety) items had dominant salient loadings on the first factor, which reflected "over-all anxiety.

cDNA cloning and expression of a novel CYP3A from the Syrian hamster, CYP3A31.

A clone, encoding a cytochrome P450 protein consisting of 501 amino acids, was isolated from a cDNA library constructed from mRNA of Syrian hamster liver. The deduced amino acid sequence of this clone showed a high homology (65 to 81%) with other mammalian CYP3As and hence, this novel isozyme was named CYP3A31. By Northern blotting, using an oligonucleotide specific to CYP3A31, the mRNA for this isozyme was shown to be expressed constitutively in liver and induced by treatment with phenobarbital but repressed by 3-methylcholanthrene or dexamethasone treatments.

Adenovirus vector-mediated gene transfer into rat retinal neurons and Müller cells in vitro and in vivo.

The ability and efficiency of human type V adenovirus-originated vector with CAG promoter to transfer foreign genes was examined in the rat retina. We introduced the adenovirus vector, AxCALacZ with the reporter gene LacZ (beta-galactosidase gene) into cultured retinal cells, and then injected the vector suspension into the vitreous cavity of the eye. Beta-galactosidase staining was observed in both glial and neuronal cells in vitro and in Müller cells near the injection site in vivo.

Cloning and characterization of Syrian hamster testosterone 7 alpha-hydroxylase, CYP2A9.

We report here the cloning of a cDNA encoding testosterone 7 alpha-hydroxylase in the Syrian hamster, designated CYP2A9. Overlapping clones encoding Syrian hamster CYP2A9 were isolated by screening a liver cDNA library and by performing rapid amplification of cDNA ends polymerase chain reaction on the cDNA library. The sequence of the CYP2A9 cDNA contains an open reading frame of 1482 nucleotides encoding a polypeptide of 493 amino acids with a calculated molecular mass of 56,295 Da.

The Japanese adaptation of the STAI Form Y in Japanese working adults--the presence or absence of anxiety.

Symptom endorsements and response patterns of 1,862 Japanese adult workers (1,509 males and 353 females) to the Japanese adaptation of the State-Trait Anxiety Inventory Form Y (STAI-JY) items, were examined in this study. The mean STAI-JY State and Trait anxiety scores of Japanese workers were substantially higher than those of American workers reported in the Manual, due primarily to the much higher scores of Japanese workers in responding to the anxiety-absent items. The correlations between the State and Trait anxiety-present scales and those of their anxiety-absent scales' counterparts were higher than those between the State anxiety-present and -absent scales and those of their Trait scales' counterparts.

Ultrasonographic assessment of regional differences in atherosclerotic lesions in patients with hypertension, diabetes mellitus, or both.

We evaluated risk factors involved in regional differences in atherosclerotic lesions in patients with hypertension, diabetes mellitus, or both. Using ultrasonography, we examined the brachial, common carotid, and common femoral arteries in 65 hospitalized Japanese patients (15 controls, 18 patients with hypertension, 16 with diabetes mellitus, and 16 with both hypertension and diabetes mellitus). They ranged in age from 39 to 81 yr, mean 60.

Effects of co-administration of butylated hydroxytoluene, butylated hydroxyanisole and flavonoids on the activation of mutagens and drug-metabolizing enzymes in mice.

Effects of co-administration of food additives and naturally occurring food components were studied on the activation of mutagens. Male mice (ddY) were given diets containing butylated hydroxytoluene (BHT) or butylated hydroxyanisole (BHA) and flavone or flavanone (2,3-dihydroflavone) for two weeks and the ability of hepatic microsomes to activate aflatoxin B1, benzo[a]pyrene and N-nitrosodimethylamine was determined by the mutagenicity test. Co-administration of an antioxidant (0.

Characterization of Orientia tsutsugamushi isolated in Taiwan by immunofluorescence and restriction fragment length polymorphism analyses.

A total of 10 strains of Orientia tsutsugamushi were isolated from field rodents and chiggers in Taiwan, and characterized by immunofluorescence analysis with monoclonal antibodies and by restriction fragment length polymorphism analysis of the 56-kilodalton (kDa) protein gene. The isolates were divided into two groups consisting of 1 and 9 strains which showed some relation to Gilliam and Karp type strains, respectively. However, all these isolates possessed characteristics distinct not only from those of known prototype strains including Gilliam and Karp but also from all isolates from Japan.

Sulfur-Bridged Cubane-Type Molybdenum-Gallium Clusters with Mo(3)GaS(4)(n)(+) (n = 5, 6) Cores. X-ray Structures of [Mo(3)GaS(4)(H(2)O)(12)](CH(3)C(6)H(4)SO(3))(5).14H(2)O and [Mo(3)GaS(4)(H(2)O)(12)](CH(3)C(6)H(4)SO(3))(6).17H(2)O.

Sulfur-bridged cubane-type molybdenum-gallium clusters [Mo(3)GaS(4)(H(2)O)(12)](5+) (2) and [Mo(3)GaS(4)(H(2)O)(12)](6+) (3) have been synthesized from [Mo(3)S(4)(H(2)O)(9)](4+) (1) and gallium metal and have been isolated as 2(pts)(5).14H(2)O (2.pts) and 3(pts)(6).

[Geriatrics in the new program for initial postgraduate medical training in Japan].

Until 1993, initial postgraduate medical training in Japan was conducted only at university hospitals or other teaching hospitals. In 1994, the number of types of training sites was expanded to include health centers, nursing homes, and extended care facilities, which gave trainees chances to understand primary medical care in various settings, providing in-home as well as other non-hospital medical services. Since the new program seemed to increase training opportunities in geriatric medicine, questionnaires regarding the new program were sent by mail to all teaching hospitals in Japan (261); 213 responded.

Organization and expression of the mouse gene for DNA repair methyltransferase.

06-Methylguanine-DNA methyltransferase (MGMT) is present in various organisms, from bacteria to human cells, and plays an important role in preventing mutations caused by alkylating substances. To understand better the regulatory mechanism involved in the expression of the gene and to construct a mouse model to investigate roles of the enzyme in carcinogenesis, the genomic sequence for mouse methyltransferase was isolated and characterized. The gene consists of 5 exons and spans over 180 kb, whereas mRNA for the enzyme was less than 1 kb.

Okadaic acid potentiates 3-methylcholanthrene-induced CYP2A8 gene expression in primary cultures of Syrian hamster hepatocytes: possible involvement of activator protein-1.

In Syrian hamster liver, treatment with 3-methylcholanthrene (3-MC) markedly induces an isozyme of cytochrome P450 (CYP), CYP2A8. To elucidate the mechanism of this induction, we studied the effect of okadaic acid (OA), an inhibitor of serine threonine protein phosphatases 1 and 2A, on 3-MC-induced CYP2A8 expression in primary cultures of Syrian hamster hepatocytes. The addition of OA to the cultured hepatocytes at a concentration of 1 nM potentiated 3-MC- (0.