Novel sterol 7 alpha-hydroxylase(s) active towards not cholesterol but side-chain oxygenated steroids in liver microsomes.
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Animals↗
Biomedical subjects
A Honda
Publications and source records attributed to A Honda.
[Polypeptide profiles of Chlamydia pneumoniae strains isolated from Japan and western blot analysis using patients' sera].
Polypeptide profiles were compared among Chlamydia pneumoniae strain of AC-43, which was isolated from a Japanese child, and other strains of C. pneumoniae. There was no distinctive differences among these strains. All strains showed marked band on 39.5 KDa area, which is equivalent with the major outer membrane protein (MOMP). Patients infected with C. pneumoniae were analyzed by Western blot using AC-43. Reaction for 98, 68, 60 KDa proteins were recognized among sera which showed positive results for anti-C. pneumoniae IgG antibody. There were two family members from whom C. pneumoniae was not isolated and anti-C. pneumoniae IgG antibody was negative. Sera obtained from them also showed weak reaction on 98 KDa protein area. Strong reaction on MOMP area was demonstrated in one patient from whom C. pneumoniae was isolated. Monoclonal antibody produced from AC-43 strain showed no reaction against homologous antigen. Reaction to 98 KDa protein was detected in all of the patients infected with C. pneumoniae. However, recognition to MOMP of C. pneumoniae seems to be different among patients. This result might suggest the presence of subtype among C. pneumoniae strains.
Blotting, Western↗
[A report of 4 cases from whom Chlamydia pneumoniae was isolated].
Chlamydia pneumoniae (C. pneumoniae) was isolated from respiratory tract of 4 girls. Case 1 is 5-year-old, diagnosed as pneumonia, had had a prolonged productive cough. She was treated with RKM with improvement of symptoms, however, C. pneumoniae was isolated repeatedly and was not deleted. Neither anti-mycoplasmal nor viral antibodies were positive and no significant pathogens were cultured from nasophrayngeal swabs. Case 2, the sister of case 1, is a 3-year-old girl with acute bronchitis treated with EM. C. pneumoniae was negative on the tenth day after treatment. Case 3, a 5-year-old girl, had a fever and was diagnosed as bronchopneumonia with a mild attack of bronchial asthma. She was initially treated with CFIX followed by therapy including EM. Her symptoms had disappeared after treatment and anti-mycoplasmal antibody was 1:320. Case 4 was an asymptomatic carrier of C. pneumoniae. Specimen was obtained at regular health examinations of junior high school. C. pneumoniae was isolated from a 14-year-old girl without respiratory symptoms. Clinical figures of C. pneumoniae infections varies from asymptomatic carrier to pneumonia. Pathogens other than C. pneumoniae could modify symptoms of infections. Precise examinations of these cases would establish a proper management of a C. pneumoniae infection.
Adolescent↗
[Cataracts in a patient treated with busulfan (Mablin powder) for eight years].
Busulfan (Mablin powder) has been used in the treatment of chronic myelogenous leukemia for a long time. Although cataract is known to be an ocular side effect of prolonged Busulfan therapy, it has rarely been reported, especially in Japan. This report describes mature cataracts in a 59-year-old woman with chronic myelogenous leukemia, who had received a total dose of 2,249.7 mg Busulfan over eight years. In our department, the average age at the time of senile cataract operation was 72 years in men and 73.4 years in women during the past five years, so this patient was younger than the average senile cataract patient. A review of case reports, including the present one, suggests that Busulfan cataract characteristically develops rapidly in both eyes of young patients treated with a total dose of over 2,000 mg of Busulfan for more than four years. Busulfan cataracts have rarely been reported probably because cataract is regarded as senile and patients treated with Busulfan usually die before cataract appears.
Busulfan↗
Manipulation of thromboxane synthesis by novel 26,27-dialkyl analogues of 1 alpha,25-dihydroxyvitamin D3 in human promyelocytic leukemia (HL-60) cells.
Using new steroidal side-chain-lengthened 26,27-dialkyl analogues of 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25-(OH)2D3], we manipulated the synthesis of thromboxane and thromboxane-producing enzymes, cyclo-oxygenase and thromboxane synthase, in human promyelocytic leukemia (HL-60) cells in serum-free culture. The order of potency of the analogues for stimulating thromboxane B2 synthetic activity from arachidonic acid (reflecting combined cyclo-oxygenase activity and thromboxane synthase activity) and from prostaglandin H2 (thromboxane synthase activity only) as well as for cyclo-oxygenase induction was 1 alpha,25-(OH)2D3 > or = 1 alpha,25-(OH)2-26,27-CH3)2D3 > 1 alpha,25-(OH)2-26,27-(C2H5)2D3 >> 1 alpha,25-(OH)2-26,27-(C3H7)2D3. These results suggest that there are functional and structural limits to the chain length of C-26 and C-27 dialkyl groups flanking the C-25-OH group in the 1 alpha,25-(OH)2D3 molecule for expressing thromboxane synthetic activity in HL-60 cells. Removal of the C-1 alpha-OH group from 1 alpha,25-(OH)2D3 led to markedly decreased thromboxane synthetic activity in HL-60 cells. These structure-activity relationships indicate that both the C-25-OH and C-1 alpha-OH groups in the 1 alpha,25-(OH)2D3 molecule are essential for expressing thromboxane synthesis in HL-60 cells. Also, the rank order for stimulating thromboxane synthesis correlated well with the binding affinity of these dialkyl analogues for the 1 alpha,25-(OH)2D3 receptor of HL-60 cells, suggesting a 1 alpha,25-(OH)2D3 receptor-mediated induction mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)
Calcitriol↗
The amino acid sequence of nucleoside diphosphate kinase I from spinach leaves, as deduced from the cDNA sequence.
The primary structure of nucleoside diphosphate (NDP) kinase from spinach leaves has been deduced from its cDNA sequence. A lambda gt 11 cDNA library derived from spinach leaves was screened using an antibody against NDP kinase I, which we previously purified to electrophoretic homogeneity (T. Nomura, T. Fukui, and A. Ichikawa, 1991, Biochim. Biophys. Acta 1077, 47-55). The cDNA sequences of positive clones contained the amino acid coding region (444 base pairs) for NDP kinase I as well as 5' and 3' noncoding regions of 33 and 361 base pairs, respectively. The cDNAs hybridized to a 1.1-kb mRNA. NDP kinase I contains 148 amino acid residues with a molecular mass of 16,305, which is in excellent agreement with that of the purified enzyme (16 kDa). Homology was found between the sequence of spinach NDP kinase I and those of the rat, Myxococcus xanthus, and Dictyostelium discoideum NDP kinases, as well as the human Nm23-gene product and the awd protein of Drosophila melanogaster.
Amino Acid Sequence↗
Mouse thromboxane A2 receptor: cDNA cloning, expression and northern blot analysis.
A cDNA clone for the mouse thromboxane A2 receptor was isolated from mouse lung cDNA library. The cDNA has a 1,023 base pair open reading frame which encodes a protein of 341 amino acid residues. STA2 and U-46619 induced inward current in Xenopus laevis oocytes injected with the transcript of the clone. Specific binding of [3H]S-145 was found in membranes of COS-1 cells transfected with the cDNA (Kd = 3.3 nM) and was displaced with unlabeled prostaglandins and thromboxane analogues in the order of S-145 greater than STA2 greater than U-46619 greater than PGD2 greater than PGF2 alpha = PGE2. Northern blot analysis demonstrated that thromboxane A2 receptor mRNA is expressed abundantly in thymus, spleen and lung.
Amino Acid Sequence↗
Cloning and expression of a cDNA for mouse prostaglandin E receptor EP3 subtype.
A functional cDNA clone for mouse EP3 subtype of prostaglandin (PG) E receptor was isolated from a mouse cDNA library using polymerase chain reaction based on the sequence of the human thromboxane A2 receptor and cross-hybridization screening. The mouse EP3 receptor consists of 365 amino acid residues with putative seven-transmembrane domains. The sequence revealed significant homology to the human thromboxane A2 receptor. Ligand binding studies using membranes of COS cells transfected with the cDNA revealed specific [3H]PGE2 binding. The binding was displaced with unlabeled PGs in the order of PGE2 = PGE1 greater than iloprost greater than PGF2 alpha greater than PGD2. The EP3-selective agonists, M&B 28,767 or GR 63799X, potently competed for the [3H]PGE2 binding, but no competition was found with EP1- or EP2-selective ligands. PGE2 and M&B 28,767 decreased forskolin-induced cAMP formation in a concentration-dependent manner in Chinese hamster ovary cells permanently expressing the cDNA. Northern blot analysis demonstrated that the EP3 mRNA is expressed abundantly in kidney, uterus, and mastocytoma P-815 cells and in a lesser amount in brain, thymus, lung, heart, stomach, and spleen.
Alprostadil↗
Effects of vitamin D-binding proteins on HL-60 cell differentiation induced by 26,26,26,27,27,27-hexafluoro-1 alpha,25-dihydroxyvitamin D.
Monocytic differentiation-inducing activity of 26,26,26,27,27,27-hexafluoro-1 alpha,25-dihydroxyvitamin D3 [26,27-F6-1 alpha,25-(OH)2D3] was re-evaluated in human promyelocytic leukemia (HL-60) cells in serum-supplemented or serum-free culture. The order of in vitro potency for reducing nitroblue tetrazolium (NBT) was 26,27-F6-1 alpha,25-(OH)2D3 greater than 1 alpha, 25-dihydroxyvitamin D3 [1 alpha,25-(OH)2D3] = 26,26,26,27,27,27-F6-1 alpha,23(S), 25-trihydroxyvitamin D3 [26,27-F6-1 alpha,23(S), 25-(OH)3D3] under serum-supplemented culture conditions, whereas the order was 1 alpha, 25-(OH)2D3 = 26,27-F6-1 alpha,25-(OH)2D3 greater than 26,27-F6-1 alpha,23(S), 25-(OH)3D3 under serum-free culture conditions. This rank order for differentiation-inducing activity under serum-free culture conditions correlated well with the binding affinity of these analogs for vitamin D3 receptor of HL-60 cells. The order of relative % binding affinity for the vitamin D-binding protein in fetal calf serum was 1 alpha,25-(OH)2D3 (100%) much greater than 26,27-F6-1 alpha,25-(OH)2D3 (5.1%) greater than 26,27-F6-1 alpha,23(S), 25-(OH)3D3 (less than 1%). These results suggest that serum vitamin D-binding proteins apparently modulate monocytic differentiation of HL-60 cells by 26,27-F6-1 alpha,25-(OH)2D3 under serum-supplemented culture conditions.
Binding, Competitive↗
Possible theophylline-minocycline interaction.
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Aged↗
Simultaneous assay of the activities of two key enzymes in cholesterol metabolism by gas chromatography-mass spectrometry.
A very sensitive and specific method for the simultaneous assay of the activities of two key regulatory enzymes in cholesterol metabolism, 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (EC 1.1.1.34), and cholesterol 7 alpha-hydroxylase (EC 1.14.13.7), is described. The assay is based on the measurement of [2H3]mevalonolactone and 7 alpha-hydroxycholesterol produced by the incubation of [2H3]HMG-CoA and endogenous cholesterol with hamster liver microsomes using isotope dilution mass spectrometry. The incubation mixture was purified by means of solid extraction cartridges, and the extract was treated with benzylamine followed by dimethylethylsilyl imidazole. The resulting ether derivatives of the mevalonylbenzylamide and 7 alpha-hydroxycholesterol were quantified by gas chromatography-mass spectrometry with selected-ion monitoring in a high resolution mode. The method made it possible to assay simultaneously the activities of HMG-CoA reductase and cholesterol 7 alpha-hydroxylase in hamster liver microsomes with high sensitivity and accuracy.
Animals↗
Microanalysis of bile acid composition in intrahepatic calculi and its etiological significance.
Brown pigment stones in the intrahepatic bile ducts were compared with those found in the extrahepatic bile ducts with special reference to the bile acids modified by bacterial intervention, that is, unconjugated, glucuronidated, secondary, and ketonic bile acid fractions. The former showed significantly lower amounts of total bile acids (P less than 0.01) and lower proportions of unconjugated bile acid fraction (P less than 0.01), secondary bile acid fraction (P less than 0.05), and ketonic bile acid fraction (P less than 0.05) to total bile acids than the latter. The discriminant analysis using these bile acid parameters led to complete separation between intrahepatic and extrahepatic stones in the case of brown pigment stones. In contrast, cholesterol stones in the intrahepatic bile ducts showed the bile acid composition close to those found in the extrahepatic ducts and gallbladder. The above data show that the bacterial infection plays a less important role in the formation and ensuing growth of most intrahepatic brown pigment stones than in extrahepatic stones, and that factors other than or in addition to bacterial infection are involved.
Bacterial Infections↗
Effects of novel 26,27-dialkyl analogs of 1 alpha,25-dihydroxyvitamin D3 on differentiation-inducing activity of human promyelocytic leukemia (HL-60) cells in serum-supplemented or serum-free culture.
Monocytic differentiation-inducing activity of steroidal side chain-lengthened 26,27-dialkyl analogs of 1 alpha,25-dihydroxyvitamin D3 was examined in human promyelocytic leukemia (HL-60) cells in serum-supplemented or serum-free culture. The order of in vitro potency for reducing nitroblue tetrazolium was 1 alpha,25-dihydroxy-26,27-dimethylvitamin D3 greater than or equal to 1 alpha,25-dihydroxy-26,27-diethylvitamin D3 much greater than 1 alpha,25-dihydroxy-26,27-dipropylvitamin D3 under serum-free culture conditions. Analysis by sucrose density-gradient centrifugation or polyethylene glycol precipitation technique showed that the potency order for differentiation-inducing activity correlated well with binding affinity of these analogs for vitamin D3 receptor of HL-60 cells. Under serum-supplemented culture conditions, the lack of correlation between biologic activity and analog-binding affinity for receptor was caused by differences in binding affinity of these analogs for serum vitamin D-binding proteins. These results suggest that serum vitamin D-binding proteins apparently modulate monocytic differentiation of HL-60 cells by these analogs under serum-supplemented culture conditions.
Calcitriol↗
[The treatment of pulmonary aspergilloma with itraconazole].
We evaluated the efficacy of Itraconazole (ITZ), a newly developed triazole antifungal agent, in 5 cases of pulmonary aspergilloma. A daily dose of 100-200 mg of ITZ was given orally for 5 to 20 months. In one patient, administration of ITZ was discontinued because of mild and transient hepatic injury. On computerized tomography and conventional roentgenogram of the chest, improvement was clearly observed in 2 of the remaining 4 cases. Symptomatic improvement, mainly disappearance of airway bleeding, was seen in 2 cases. Thus the overall improvement rate was 50% (two out of 4). No adverse response other than the hepatic injury mentioned above occurred. In one patient, who had two fungus balls previously, one of the fungus balls disappeared completely and the other decreased significantly in size during the course of treatment. In this case, the daily dose of ITZ was larger (200 mg once a day) and the plasma concentration of ITZ was extremely higher (2068 ng/ml) than in other cases (137-330 ng/ml). We conclude that ITZ is an efficacious drug in the treatment of pulmonary aspergilloma, and that this drug may be even more efficacious if we keep higher plasma concentration of ITZ.
Adult↗
Cooperative enhancement of hyaluronic acid synthesis by combined use of IGF-I and EGF, and inhibition by tyrosine kinase inhibitor genistein, in cultured mesothelial cells from rabbit pericardial cavity.
We examined the combined effects of IGF-I and EGF on hyaluronic acid synthesis using cultured pericardial mesothelial cells from rabbit pericardial cavity. Combined exposure of pericardial cells to IGF-I and EGF cooperatively increased hyaluronic acid synthesis and the level of hyaluronic acid synthase activity over that seen with or without IGF-I or EGF alone, but did not significantly stimulate the sulfated glycosaminoglycan synthesis. Increase in the level of hyaluronic acid synthesis and hyaluronic acid synthase activity induced by these growth factors was blocked after pretreatment with a tyrosine-specific protein kinase inhibitor, genistein. Genistein had no direct inhibitory effect on hyaluronic acid synthase activity. These results suggest that cooperative enhancement of hyaluronic acid synthesis induced by combined use of IGF-I and EGF could be mediated by a receptor tyrosine kinase-involved transmembrane signaling process that is responsive to IGF-I and EGF.
Animals↗
Effect of marine coral prostanoids, clavulones, on spontaneous beating rate of cultured myocardial cells from fetal mouse hearts.
We examined effects of newly discovered marine coral prostanoids, clavulones, isolated from the Japanese stolonifer Clavularia viridis, on the spontaneous beating rate of cultured myocardial cells from fetal mouse hearts. Clavulone caused positive chronotropic action at 2-5 min after addition of clavulone (0.45 microM) to the reaction media of cardiac cells. This effect induced by clavulone was clearly different from the positive inotropic effects of ouabain (10 microM) and Bay K 8644 (0.1 microM) as judged by photoelectric recordings of beating. These results suggest a new biological action of clavulone that has positive chronotropic action on the cultured mouse myocyte preparation.
3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
[The defensive role of human pulmonary alveolar macrophages and polymorphonuclear leukocytes against a strongly or weakly virulent strains of Cryptococcus neoformans].
The phagocytic and killing activities of human pulmonary alveolar macrophages (PAM) and polymorphonuclear leukocytes (PMN), and superoxide (O2-) production in phagocytizing PAM and PMN were studied by using a strongly and weakly virulent strains of Cryptococcus neoformans (C. neoformans) in mice. The strongly virulent strain was encapsulated with a thick capsule while the other had a thin one. The phagocytic and killing activities of PAM and PMN against the strongly virulent strain were lower than against the weakly virulent one. There was no significant difference between PAM and PMN in terms of phagocytic or killing activities and O2- production. It seems likely that O2- plays no defensive role in the intracellular killing of C. neoformans by PAM and PMN because no correlation was found between the killing activity and O2- production. These results indicate that human PAM and PMN play an effective role in host defense against the weakly virulent strain of C. neoformans encapsulated with a thin capsule but are not effective against the strongly virulent strain encapsulated with a thick capsule.
Adolescent↗
The biochemical and pharmacological manipulation of cellular cyclooxygenase (COX) activity.
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Animals↗