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M Noshiro

Publications and source records attributed to M Noshiro.

At least 73 records · Page 4Linked to original sources
PubMed1994

Rat hepatic 3 alpha-hydroxysteroid dehydrogenase: expression of cDNA and physiological function in bile acid biosynthetic pathway.

3 alpha-Hydroxysteroid dehydrogenase (3 alpha-HSD) [EC 1.1.1.213]2 plays important multifunctional roles in metabolizing steroid hormones, polycyclic aromatic hydrocarbons, and prostaglandins and also in transforming the steroid nucleus for the biosynthesis of bile acids from cholesterol in liver. To gain insight into the details and physiological functions of 3 alpha-HSD in the bile acid biosynthetic pathway, cDNA clones of 3 alpha-HSD were isolated from rat liver lambda phage cDNA libraries by using specific antibodies to 3 alpha-HSD purified from rat liver. Transfection of the 3 alpha-HSD cDNA in Simian COS7 cells resulted in the expression of an immunoreactive protein to the antibodies against the purified enzyme, and the transfected cells exhibited activities for not only 7 alpha-hydroxy-5 beta-cholestan-3-one, the intermediate of bile acid biosynthesis, but also steroid hormones and 9,10-phenanthrenequinone. Northern blot analysis on poly(A)+ RNA by selective use of different cDNA fragments of the 5'-untranslated region, the coding region, and the 3'-untranslated region as probes revealed three hybridizable bands, 3.6, 2.7, and 2.5 kb, in liver and four bands, 3.6, 2.7, 2.5, and 1.8 kb, in ovary. Of these, the 2.7- and 1.8-kb bands were predominant in liver and ovary, respectively. Northern hybridization analysis also revealed that the coding region of the various sizes of mRNA seemed to be common. Southern blot analysis of genomic DNA by the selective use of the cDNA fragments as probes indicated that the various mRNA species were derived from a single gene, probably due to an alternative splicing mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Hydroxysteroid Dehydrogenases↗
PubMed1994

Transforming growth factor-beta-1 reduces alkaline phosphatase mRNA and activity and stimulates cell proliferation in cultures of human pulp cells.

Transforming growth factor-beta-1 (TGF-beta-1) is a potent modulator of proliferation and differentiation in various tissues, and may be involved in the control of dental development and repair. This study was carried out to investigate the effects of TGF-beta-1 on alkaline phosphatase (ALPase) activity and mRNA level, and on DNA content in cultures of human pulp cells. Four lines of pulp cells (P1-P4), isolated from the upper wisdom teeth of four patients, were maintained separately in monolayer cultures in the presence of 10% fetal bovine serum. TGF-beta-1, at 0.1 ng/mL, increased ALPase activity and DNA content in P1 cultures, but not in P2-P4 cultures. In all cultures, TGF-beta-1, at 5 ng/mL, decreased ALPase activity to a very low level, and increased DNA content. Northern analysis showed that human pulp cells synthesized a single species of 2.6-kb liver/bone/kidney-type ALPase, and that TGF-beta-1, at 5 ng/mL, decreased the level of the ALPase mRNA. These results suggest that TGF-beta-1 is a mitogen for human pulp cells, and that it regulates the activity of the universal-type ALPase at the pre-translational level.

Alkaline Phosphatase↗
PubMed1994

Regulation of messenger ribonucleic acid expression of 1 alpha,25-dihydroxyvitamin D3-24-hydroxylase in rat osteoblasts.

We have reported that PTH inhibits 25-hydroxyvitamin D3-24-hydroxylase messenger RNA (mRNA) expression induced by 1 alpha,25-dihydroxyvitamin D3 [1 alpha, 25-(OH)2D3] in rat kidney but not intestine. In the present study, we examined whether the suppression of 24-hydroxylase mRNA expression by PTH occurs commonly in tissues and cells which have PTH receptors. Administration of 1 alpha, 25-(OH)2D3 into rats fed a synthetic vitamin D-repleted diet containing adequate calcium greatly increased serum levels of calcium and 1 alpha, 25-(OH)2D3. Also, there was a 4-fold increase in bone 24-hydroxylase activity in response to 1 alpha, 25-(OH)2D3 administration. In rats fed a low calcium diet, renal 24-hydroxylase activity was suppressed probably due to secondary hyperparathyroidism. In contrast, the low calcium feeding did not suppress bone 24-hydroxylase activity. The expression of 24-hydroxylase mRNA in rat osteoblastic C-26 and C-11 cells was similar and attained maximal levels 24 h after cells were incubated with 10(-8) M 1 alpha, 25-(OH)2D3. Induction of 24-hydroxylase mRNA expression by 1 alpha, 25-(OH)2D3 was much greater and earlier in immature C-26 cells than mature C-11 cells. Simultaneous addition of PTH, prostaglandin E2, or cAMP together with 1 alpha, 25-(OH)2D3 did not down-regulate mRNA expression of 24-hydroxylase induced by the vitamin in both C-26 and C-11 cells. Of the three osteoblastic cells (C-26, C-20, and C-11) examined, C-26 cells showed the least mRNA expression of vitamin D receptors, in spite of the highest expression of 24-hydroxylase mRNA. These results suggest that unlike in the kidney, bone 24-hydroxylase mRNA expression is not down-regulated by PTH despite of the presence of PTH receptors. They also suggest that the degree of the induction of 24-hydroxylase mRNA by 1 alpha, 25-(OH)2D3 is not explained simply by the vitamin D receptors content.

Adenylyl Cyclases↗
PubMed1994

[Analysis of middle ear cavity gas composition by mass spectrometry].

Gas compositions in the middle ear cavities of patients with a perforated tympanic membrane, and in normal subjects, were analyzed by mass spectrometry. The results showed that the gas in normal subjects consisted of nitrogen (83.7%), oxygen (9.2%) and carbon dioxide (6.2%). (The volume concentration of the various gases was expressed as a percentage of 713 mm Hg.) Thus, the concentrations of the various gases in the middle ear cavity differed greatly from those in the atmosphere. The low concentration of oxygen is suitable for microaerophilic bacteria and acts as a physiological barrier against bacterial infection. It was also found that the gas in patients with exudative otitis media consisted of nitrogen (77.9%), oxygen (15.6%) and carbon dioxide (5.5%), and in patients with chronic perforated otitis media, of nitrogen (77.8%), oxygen (16.9%) and carbon dioxide (4.4%). The concentration of oxygen in the middle ear cavity with perforated tympanic membrane is higher than that in the normal state because of diffusion of atmospheric gas into the middle ear cavity. In other words, perforation of the tympanic membrane exposes the middle ear cavity to a nonphysiological gaseous condition. The diffusion of atmospheric gas was simulated using small cylinders. Simulated results revealed that the atmospheric gas diffused rapidly. The change in the gas concentration during diffusion was approximated by an exponential function and a constant, and reproduced by a mathematical model. The change in the gas concentration in the middle ear cavity was usually approximated by two exponential functions and a constant. The constant represents the estimated final value of the gas concentration from which the effect of the diffusion is excluded. The final values were estimated for normal subjects and patients with perforated tympanic membrane. The final values in the normal subjects were almost equal to the measured values. The final value of carbon dioxide in the patients was greater than the measured value and nearly equal to the final value in the normal subjects. The final oxygen value in the patients was smaller than the measured value, but still greater than the final value in the normal subjects. The high oxygen concentration in the patients suggests enhanced oxygen metabolism.

Adult↗
PubMed1993

Structure of the gene encoding human liver cholesterol 7 alpha-hydroxylase.

A human cholesterol 7 alpha-hydroxylase gene spanning 17 kb in length which includes the sequence 5' flanking region (6. kb) and that of the entire transcriptional region (10.5 kb) was obtained from two partially overlapping clones (i.e., HG 18 and HG 17) of a human genomic library. The exon-intron boundaries are completely identical with that of rat gene. Several transcriptional factor recognition sequences were observed in the 5' flanking region.

Amino Acid Sequence↗
PubMed1993

Structural characterization of the gene encoding rat 25-hydroxyvitamin D3 24-hydroxylase.

The structural gene encoding 25-hydroxyvitamin D3 24-hydroxylase (P-450cc24) was isolated from the rat genomic DNA. It spans approximately 15 kb, is composed of 12 exons, and was demonstrated by Southern blot analysis to be present as a single copy. One major T residue was identified at the cap site, a putative TATA (ATAAATA) box was located at position -30, and a putative CCAAT box was at -58. Four possible vitamin D responsive elements that may be involved in regulation of 24-hydroxylase expression were found in the 5'-flanking region. Alignment with mitochondrial P-450 proteins showed that 7 out of 11 intron insertion sites of P-450cc24 gene occupied positions identical with those in the CYP11 family (P-450scc, P-450(11 beta)). The structure of the gene is discussed in relation to present knowledge about the mechanism of regulation of the 25-hydroxyvitamin D3 24-hydroxylase and calcium homeostasis.

Amino Acid Sequence↗
PubMed1993

Electrical impedance in the lower limbs of patients with Duchenne muscular dystrophy: a preliminary study.

Electrical impedances were measured, at a range of frequencies from 2 to 200 kHz, in the lower limbs of nine patients with Duchenne muscular dystrophy and 12 normal subjects. The specific admittance was calculated from the impedance, based on the assumption that the shape of the part of the limb measured is a truncated cone. Curve-fitting to the admittance locus on the complex plane gave the intracellular and extracellular resistances and the membrane capacitance. The intracellular resistance was higher and the membrane capacitance was lower in the patients than in the normal subjects. The membrane capacitance in the normal subjects increased with age. In the patients, the membrane capacitance decreased, and the intracellular resistance increased with the stage of disability, which was assessed by a physician. These experimental results agree well with the theoretical explanation, which uses simplified spherical models to represent living tissue.

Adolescent↗
PubMed1993

Nonlinear identification of the PCO2 control system in man.

Two approaches to identification of the PCO2 system in man are described. The first uses a nonlinear 'black box' NARMAX identification package, while the second method uses a structured two-compartment Belville model. The data were obtained from volunteers breathing either room air or a controlled gas mixture, controlled via a pseudorandom M-sequence. Measurements were made of respiratory gas flow and PCO2 content of inspired and expired gases. The identification results indicate that a low-order dynamic model with nonlinear polynomial expansion gave the best fit to the data. In contrast, the Belville model gave best results with a two-compartment linear model, mainly because of difficulties in the optimisation routines when the Belville model was not linear. Thus, modern systemic methods of excitation and identification appear to be appropriate for modelling this respiratory subsystem of humans.

Adult↗
PubMed1993

Effects of interleukin-1 on syntheses of alkaline phosphatase, type X collagen, and 1,25-dihydroxyvitamin D3 receptor, and matrix calcification in rabbit chondrocyte cultures.

The effect of IL-1 on expression of the mineralization-related phenotype by chondrocytes was examined. In cultures of rabbit growth plate chondrocytes, IL-1 beta at 0.1 ng/ml caused 95% decreases in alkaline phosphatase activity, alkaline phosphatase mRNA levels, the incorporation of 45Ca into insoluble material, and the calcium content during the hypertrophic stage. These effects of IL-1 beta were dose-dependent and were observed in 24-48 h. Furthermore, IL-1 beta suppressed increase in cell size and the syntheses of 1,25-dihydroxyvitamin D3 receptor and type X collagen, other markers of hypertrophy, but had little effect on the synthesis of total protein including type II collagen. The inhibition of calcification was observed only when chondrocytes were exposed to IL-1 before the onset of calcification: IL-1 treatment from the mineralization stage had a marginal effect on 45Ca incorporation into insoluble material. These results suggest that IL-1 inhibits chondrocyte hypertrophy and the onset of calcification in ossifying cartilage.

Alkaline Phosphatase↗
PubMed1993

Partial cloning of cDNA for hamster liver cholesterol 7 alpha-hydroxylase.

A partial cDNA clone encoding cholesterol 7 alpha-hydroxylase (pHA 7 alpha-3) was isolated from a hamster liver cDNA library using rat P-450 ch7 cDNA (p 7 alpha-11) as a probe. In comparison with the complete base sequence of p 7 alpha-11, pHA 7 alpha-3 lacked about 500 base pairs at the 5' end. Circadian rhythms of mRNA for hamster liver 3-hydroxy-3-methylglutaryl coenzyme A reductase and cholesterol 7 alpha-hydroxylase were investigated with cDNA clones encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase (pRed-227) and pHA 7 alpha-3. Typical circadian rhythms of mRNA for both enzymes and a time lag between the two circadian rhythms were observed. The circadian rhythm of activity for each enzyme corresponded to that of each mRNA.

Animals↗
PubMed1992

Purification and characterization of 7 alpha-hydroxy-4-cholesten-3-one 12 alpha-hydroxylase.

The isoform of cytochrome P450 that catalyzes the 12 alpha-hydroxylation of 7 alpha-hydroxy-4-cholesten-3-one, an intermediate in the conversion of cholesterol to cholic acid, was purified to homogeneity from rabbit liver microsomes. The extent of purification in the various steps was judged by an assay involving high performance liquid chromatography. The purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis (M(r) = 50,000). The NH2-terminal amino acid sequence is as follows: Val-Leu-Trp-Gly-Leu-Leu-Gly-Ala-Leu-Leu-Met-Val-Met-Val-Gly-, which is different from that of any other P450s so far reported. The specific content of the enzyme was 13.3 nmol of cytochrome P450/mg of protein. Upon reconstitution with NADPH-cytochrome P450 reductase and cytochrome b5, the P450 enzyme showed a high activity of 12 alpha-hydroxylation with a turnover number of 36.6 min-1 at 37 degrees C. The omission of either cytochrome P450 or NADPH-cytochrome P450 reductase resulted in complete loss of activity, and the omission of cytochrome b5 resulted in 40% loss of activity. Antibodies prepared from mouse inhibited the 12 alpha-hydroxylase activity of rabbit liver microsomes about 90% and that of the rat liver microsomes 50%. The enzyme activity was not inhibited by other antibodies raised against other forms of P450 that catalyze different monooxygenation reactions toward xenobiotics or endogenous substrates. Anti-cytochrome b5 antibody inhibited the activity 40%, suggesting the functional role of this protein, and anti-reductase inhibited the activity almost completely. The microsomal enzyme activity was markedly elevated by starvation or streptozotocin administration to the animals. However, an immunoblotting experiment showed no correlation between the enzyme activity and the amount of protein, suggesting that post-translational modification may occur.

Amino Acid Sequence↗
PubMed1992

Parathyroid hormone inhibits 25-hydroxyvitamin D3-24-hydroxylase mRNA expression stimulated by 1 alpha,25-dihydroxyvitamin D3 in rat kidney but not in intestine.

Using a cDNA probe for rat renal 24-hydroxylase, expression of its mRNA was compared in the rat kidney and intestine. Vitamin D-deficient rats received a single injection of 1 alpha,25-dihydroxyvitamin D3. Expression of 24-hydroxylase mRNA was first detected in the kidney at 3-h post-injection and increased thereafter. Similarly, 24-hydroxylase mRNA was expressed in the intestine after 1 alpha,25-dihydroxyvitamin D3 injection. However, the dose level of 1 alpha,25-dihydroxyvitamin D3 required to induce the intestinal 24-hydroxylase mRNA expression was only 1/100 the amount required to induce renal 24-hydroxylase mRNA. Induction of intestinal 24-hydroxylase mRNA expression by 1 alpha,25-dihydroxyvitamin D3 was far more rapid than that of renal 24-hydroxylase mRNA. Thyroparathyroidectomy shortened the time required to induce expression of renal, but not intestinal, 24-hydroxylase mRNA. Administration of either parathyroid hormone or cAMP to vitamin D-deficient rats greatly reduced the expression of 24-hydroxylase mRNA in the kidney but not in the intestine. When rats were fed a vitamin D-repleted diet containing 0.7% (adequate) or 0.03% (low) calcium for 2 weeks, intestinal expression of 24-hydroxylase mRNA could be induced only in the low calcium group. In contrast, renal mRNA expression was preferentially stimulated in the adequate calcium group. These results clearly demonstrate that the expression of 24-hydroxylase mRNA is down-regulated by parathyroid hormone in the kidney but not in the intestine.

Animals↗
PubMed1992

[Molecular biology of cytochrome P450 in kidney].

Cytochrome P450-catalyzed monooxygenation system is widespread in nature and many isoforms have been found in almost all the tissues of vertebrates. In the kidney, several isoforms of cytochrome P450 families, CYP1, 2, 3, 4, 24, 27 have been characterized by biological, immunochemical and molecular biological techniques. These cytochrome P450 isoforms are localized in the endoplasmic reticulum or mitochondria of kidney cortex and functional in the monooxygenation of a number of substrates, such as steroids, fatty acids, xenobiotics, and vitamin D. Particularly, hydroxylation of vitamin D at positions of 1 and 24 is known to be one of the most important functions of kidney specific P450 isoforms.

Animals↗
PubMed1991

Molecular cloning and sequence analysis of cDNA encoding delta 4-3-ketosteroid 5 beta-reductase of rat liver.

A cDNA clone encoding delta 4-3-ketosteroid 5 beta-reductase was isolated from rat liver cDNA libraries using antibodies specific for the enzyme and oligonucleotides as probes. The cDNA contained 981-base pair open reading frame encoding 327 amino acid residues (Mr 37,376) and an unusually long 3'-untranslated region rich in AT sequence in the total length of 3189 base pairs. The predicted amino acid sequence contains the sequences similar to the putative NADPH- and steroid-binding regions.

Amino Acid Sequence↗
PubMed1991

Structural analysis of the gene encoding rat cholesterol alpha-hydroxylase, the key enzyme for bile acid biosynthesis.

The gene encoding cholesterol 7 alpha-hydroxylase (P450VIIA) was isolated from rat genomic DNA. The gene spanned about 11 kilobases and contained six exons. Blotting analysis of genomic DNA and complete matching of restriction maps of several isolated genomic clones indicated that there appeared to be only one gene in the rat genome. The putative transcription initiation site was present 61 base pairs upstream from the ATG codon. The typical TATA sequence and CCAAT promoter element were found at 24 and 47 base pairs upstream from the transcription initiation site, respectively. Alignment of several P450 proteins showed that the cholesterol 7 alpha-hydroxylase gene shared location of introns with none of the other P450 genes except for intron 5, which was in the same position as intron 10 of the gene encoding P450IVA1. The alignment also indicated that the distal helix of cholesterol 7 alpha-hydroxylase contained an asparagine in place of the well conserved threonine that is postulated to be involved in the O2 binding site. Unusual residues, Asn-126 and Thr-442, were also found at the sites where all other P450s have positively charged amino acids, which are considered to be involved in interaction with heme propionate. These replacements may be related to the unique function and unusual lability of the hydroxylase. Analysis of evolutionary distance between the cholesterol 7 alpha-hydroxylase gene and other known P450 genes indicated that yeast P450LIA is most closely related to P450VIIA. This finding suggests that the cholesterol 7 alpha-hydroxylase gene is an evolutionarily old P450 gene.

Amino Acid Sequence↗
PubMed1991

Expression of rat liver vitamin D3 25-hydroxylase cDNA in Saccharomyces cerevisiae.

The cDNA coding for the precursor protein of rat liver mitochondrial vitamin D3 25-hydroxylase, cytochrome P450LMT25, was expressed under the control of the yeast alcohol dehydrogenase I promoter and terminator in Saccharomyces cerevisiae AH22 cells. The transformed yeast cells produced a P450LMT25 protein with an almost similar apparent molecular weight as compared with that of the authentic mature enzyme. The expression level of the P450LMT25 hemoprotein was about 5 x 10(4) molecules per cell as determined by reduced CO-difference spectra. The mitochondrial fraction prepared from the transformed yeast cells exhibited both 25-hydroxylase activity toward 1 alpha-hydroxyvitamin D3 and 27-hydroxylase activity toward 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol in a reconstituted system containing bovine adrenodoxin and NADPH-adrenodoxin reductase.

Alcohol Dehydrogenase↗
PubMed1991

Structures and characterization of sex-specific mouse cytochrome P-450 genes as members within a large family. Duplication boundary and evolution.

We characterized two female-specific testosterone 16 alpha-hydroxylase mouse cytochrome P-450 genes, 16 alpha oh-a and 16 alpha oh-b. Gene 16 alpha oh-a, consisting of nine exons, is approximately 38 kbp in size. The exon sequence of this P-450 gene is identical to cDNA pf26 nucleotide sequence [Noshiro, M., Lakso, M., Kawajiri, K. & Negishi, M. (1988) Biochemistry 27, 6434-6443], which encodes female-specific testosterone 16 alpha-hydroxylase regulated by the murine Rip locus. Gene 16 alpha oh-b, containing nine exons with the same junctions as the 16 alpha oh-a, spans at least 20 kbp, and encodes a cytochrome P-450 whose deduced amino acid sequence is 90% similar to the hydroxylase. Nucleotide sequences revealed that duplication of the two genes occurred 4-22 million years ago, and that the 5' duplication boundary is located 1336 bp upstream from the putative transcription-start site. In the flanking regions of both genes, there is a long stretch (100 bp) of CA repeats in addition to other motifs, including TATA box, glucocorticoid-response-element-core and Simian-virus-40-enhancer sequences and IgG light-chain gene promoter. We isolated many genomic DNA clones which contain exon 1 sequences, and compared their restriction maps, cross-hybridization and nucleotide sequences. The results indicate that these genomic clones represent closely related genes in the 16 alpha oh family with a minimum of 16 members, which is further divided into classes a, b and c. 16 alpha oh-a and 16 alpha oh-b belong to the first and second classes, respectively. Moreover, extensive segmental gene conversion and nonreciprocal recombination were noted among the genes, particularly among those in class b. All genes in that class contain the long ATTT repeat sequences in intron 1, which may have triggered a rapid gene conversion and/or stabilize the duplicated genes.

Animals↗