PubMed Health⌕ Search

Biomedical subjects

K Iba

Publications and source records attributed to K Iba.

At least 55 records · Page 3Linked to original sources

Glucocorticoids induce mineralization coupled with bone protein expression without influence on growth of a human osteoblastic cell line.

Mineralization was induced by glucocorticoid treatment in a human osteoblastic cell line derived from normal bone in vitro, designated SV-HFO, immortalized with simian virus 40 (CHIBA, H. et al. (1993). Jpn. J. Cancer Res., 84: 290-297). Mineralization was revealed by electron microscopy, von Kossa staining and electron spectroscopic analysis, which indicated that the Ca/P ratio was approximately 1.70, corresponding to the value of hydroxyapatite. The effect was dose- (10(-8)-10(-6) M) and time-dependent (days 7-28), was greatest at day 28, and was preceded by expression of alkaline phosphatase (ALP) and osteopontin (OPN). The ALP activity induced was highest at day 7, whereas OPN reached its highest level at day 28. When the induction of ALP activity was inhibited by 10(-4) M levamisole, mineralization of SV-HFO cells by glucocorticoid treatment was markedly reduced, suggesting that elevated ALP activity in the early phase is important in the mineralization of human osteoblastic cells. Glucocorticoid treatment did not alter cell proliferation. These results indicated that glucocorticoids play crucial roles in the formation of mineralized matrix in human osteoblasts by inducing differentiation of SV-HFO cells without modulating their proliferative activity.

Alkaline Phosphatase↗

Cloning of a cDNA encoding tobacco omega-3 fatty acid desaturase.

A cDNA was isolated from a tobacco (Nicotiana tabacum cv. SR1) leaf cDNA library using, as a hybridization probe, a cDNA fragment from the gene (fad7) encoding Arabidopsis thaliana chloroplast omega-3 fatty acid (FA) desaturase. The deduced 379-amino-acid (aa) sequence has 67-71% identity to those deduced from the previously described genes encoding the omega-3 FA desaturases of A. thaliana. The absence of an N-terminal extension transit peptide in the deduced aa sequence of the cDNA clone and the accumulation pattern of the mRNA corresponding to this cDNA in leaf and root tissues indicate that the isolated cDNA encodes a tobacco microsomal omega-3 FA desaturase.

Amino Acid Sequence↗

Quantitative determination of F6-1,25(OH)2 vitamin D3 in human serum by gas chromatography/mass spectrometry with high-resolution selected ion monitoring.

A highly sensitive and specific gas chromatographic/mass spectrometric method was developed for the determination of very low levels of F6-1,25(OH)2 vitamin D3, a new fluoro analogue of vitamin D3, to facilitate investigation of its pharmacokinetics in man. Deuterium-labelled F6-1,25(OH)2 vitamin D3 was employed as an internal standard for quantification. The method involves extraction from serum samples, separation by a cartridge column system followed by straight-phase high-performance liquid chromatography, O-trimethylsilylation, and analysis by gas chromatography/mass spectrometry using high-resolution selected ion monitoring. With a quantification limit of 2 pg ml-1 and a requirement for 2 ml of serum, the assay method has sufficient sensitivity, selectivity and precision for the analysis of very low concentrations of the drug.

Calcitriol↗

Cloning of a temperature-regulated gene encoding a chloroplast omega-3 desaturase from Arabidopsis thaliana.

Previous genetic evidence suggested that the fad8 and fad7 genes of Arabidopsis thaliana encode chloroplast membrane-associated omega-3 desaturases. A putative fad8 cDNA was isolated by heterologous hybridization using a gene encoding an endoplasmic reticulum-localized omega-3 desaturase (fad3) as a probe. The cDNA encodes a protein of 435 amino acid residues with a molecular mass of 50,134 D. Constitutive expression of the cDNA in transgenic plants of a fad7 mutant resulted in genetic complementation of the mutation, indicating that the fad7 and fad8 gene products are functionally equivalent. Expression of the fad8 cDNA in transgenic plants often resulted in the co-suppression of both the endogenous fad7 and fad8 genes in spite of the fact that these two genes share only about 75% nucleotide identity. In contrast to all other known plant desaturases, including fad7, the steady-state level of fad8 mRNA is strongly increased in plants grown at low temperature. This suggests that the role of fad8 is to provide increased omega-3 desaturase activity in plants that are exposed to low growth temperature. The fad8-1 mutation created a premature stop codon 149 amino acids from the amino-terminal end of the fad8 open reading frame, suggesting that this mutation results in a complete loss of fad8 activity.

Amino Acid Sequence↗

Hormonal regulation of connexin 43 expression and gap junctional communication in human osteoblastic cells.

We have recently shown that connexin 43 (Cx43), a major gap junction protein in osteoblasts, is expressed with an increase in cell density (CHIBA, H. et al. (1993). Cell Struc. Funct., 18: 419-426). In the present study, we examined what kinds of hormones and cytokines regulate the gap junction protein in osteoblastic cells, using a human osteoblastic cell line (SV-HFO) after reaching a confluent density to avoid influence of cell proliferation. Either retinoic acid (RA) or transforming growth factor-beta 1 (TGF-beta 1) induced the Cx43 expression of SV-HFO cells, as revealed by Northern blot analysis and immunocytochemistry. These modulators also increased gap junctional intercellular communication, in terms of the extent of dye transfer. On the other hand, 1 alpha, 25-dihydroxyvitamin D3 did not influence the Cx43 expression and gap junctional intercellular communication of the cells. These results suggest that RA and TGF-beta might maintain bone tissue as an organized tissue in vivo by increasing intercellular communication of osteoblastic cells.

Calcitriol↗

A gene encoding a chloroplast omega-3 fatty acid desaturase complements alterations in fatty acid desaturation and chloroplast copy number of the fad7 mutant of Arabidopsis thaliana.

Mutations at the fad7 locus of Arabidopsis thaliana (previously called fadD) cause decreased desaturation of dienoic fatty acids in chloroplast lipids in plants grown at elevated temperatures. This suggested that the fad7 locus encodes a chloroplast omega-3 desaturase that catalyzes the desaturation of lipid-linked 18:2 and 16:2 fatty acids. In order to clone the fad7 gene, it was first genetically mapped relative to the flanking restriction fragment length polymorphism markers 4547 and 2488A on chromosome 3, and yeast artificial chromosomes covering the locus were identified. A putative desaturase cDNA clone that was isolated by low stringency heterologous probing with a cDNA for an endoplasmic reticulum-localized omega-3 desaturase (fad3) hybridized to the yeast artificial chromosomes and could not be resolved from the locus by restriction fragment length polymorphism mapping. Expression of the cDNA in transgenic fad7 mutant plants resulted in restoration of wild type fatty acid composition and suppression of a previously observed effect of the fad7 mutation on chloroplast number, indicating genetic complementation. The structural gene contained seven introns within a coding sequence of 1338 base pairs, which encodes a 446-amino acid polypeptide of 51,172 daltons. The amino-terminal region of the fad7 gene product contained a consensus chloroplast transit peptide. Except for the amino-terminal domain, the deduced amino acid sequence of the fad7 gene product had high homology to the fad3 gene product, indicating that fad7 encodes an omega-3 desaturase and that the two genes arose from a common ancestral gene. There was no apparent effect of growth temperature on the steady-state levels of fad7 mRNA in wild type plants.

Amino Acid Sequence↗

Metabolism of a tetrahydroaminoacridine derivative (SM-10888) in rat: structural analysis of an N-glucuronide of SM-10888 and an O-glucuronide of hydroxylated SM-10888 by FAB-MS/MS.

1. The metabolism of 9-amino-8-fluoro-1,2,3,4-tetrahydro-2,4-methanoacridine citrate (SM-10888), a cholinesterase inhibitor was studied in rat. 2. The phase I metabolite (designated M3) was isolated from urine and identified as 1-hydroxylated SM-10888 by 1H-n.m.r. and EI-MS. 3. Two glucuronides (designated SMG and M3G) were isolated from bile and urine and their structures examined by FAB-MS/MS and beta-glucuronidase hydrolysis. 4. FAB-mass spectra of SMG and M3G showed molecular ions ([M+H]+) at m/z 405 and 421, respectively. In their daughter spectra, fragment ions of aglycones (SM-10888 and M3), generated by the loss of glucuronic acid (176 amu) were observed. The daughter spectra of these aglycones were essentially similar to those of the corresponding synthetic standards. 5. SMG was hydrolysed non-enzymically at pH 5 as is often the case with N-glucuronides of arylamines. M3G could be hydrolysed by beta-glucuronidase but proved stable at pH 5. 6. From these results, SMG and M3G were concluded to be the N-glucuronide of SM-10888 and the O-glucuronide of M3, respectively.

Aminacrine↗

Identification and map position of YAC clones comprising one-third of the Arabidopsis genome.

YAC clones corresponding to 125 Arabidopsis thaliana RFLP markers have been identified. At least one YAC clone has been isolated for each of the RFLP markers tested. Based on CHEF gel analysis of 196 clones, the mean insert size of the available Arabidopsis YAC libraries is approximately 160 kb. The YACs of known genetic map location encompass about 30% of the Arabidopsis genome. The results presented here represent a first step towards assembly of an overlapping YAC library of the A. thaliana genome.

Arabidopsis↗

[A case of idiopathic aseptic osteonecrosis of the patellae: repeat study of Tc-99m HMDP bone scintigraphy in a reparative phase].

A 37-year-old woman entered our hospital because of bilateral knee pain. Tc-99m HMDP bone scintigram demonstrated increased activity in the bilateral patellae and cold lesion in the center and left lateral segments of the left patellae, so-called doughnut pattern. Trephine biopsy was performed to prove bilateral idiopathic asepct osteonecrosis of the patellae. The knee pain subsided and a bone scan 5 months later demonstrated increased activity in the bilateral patellae more widely than initial scan. We report the usefulness of Tc-99m HMDP bone scintigraphy to observe a reparative phase of idiopathic asepct osteonecrosis of the patellae.

Adult↗

[Reverse occlusion diagnosed following angle SN to FH correction].

A 12-year-old boy was first seen with the chief complaint of reverse occlusion. His records allowed a class III malocclusion, crown fracture of 1 angle and -1.1 degrees angle SN to FH. Therefore, we corrected angle SN to FH from -1.1 degrees to 7 degrees and treated him with intermaxillary anchorage appliance and by the edgewise method.

Child↗

[Case report of maxillary protrusion].

The patient, a 10-years and 7-month-old boy, demonstrated maxillary protrusion (a pseudo class I malocclusion) complicated by horizontal open bite, space defect of 5 5, and represented that point A was +3.0 mm, pogonion was -8.0 mm to McNamara line respectively. His 4 first premolars were extracted and he was treated with an edgewise appliance together with a high pull head gear, palatal bar, class III elastics, class II elastics and up-down elastics for 3 years. A functional occlusion and good profile was obtained with good growth, chin control and tooth movement.

Child↗

Roles of bacteriochlorophyll and carotenoid synthesis in formation of intracytoplasmic membrane systems and pigment-protein complexes in an aerobic photosynthetic bacterium, Erythrobacter sp. strain OCh114.

Synthesis of bacteriochlorophyll and carotenoids was inhibited in an aerobic photosynthetic bacterium, Erythrobacter sp. strain OCh114, by alpha, alpha'-dipyridyl and diphenylamine. Formation of two pigment-protein complexes, reaction center-B870 (RC-B870) and B806, and development of the intracytoplasmic membranes of the cells were studied by spectral analysis and electron microscopy. Inhibition of bacteriochlorophyll synthesis by alpha, alpha'-dipyridyl, which was accompanied by a decrease in carotenoid synthesis, suppressed formation of intracytoplasmic membranes in the cells. Growth under illumination had a similar effect on formation of pigments and membranes. On the other hand, inhibition of carotenoid synthesis by diphenylamine did not suppress either development of the membrane system or bacteriochlorophyll synthesis. Formation of RC-B870 and B806 complexes, however, was differentially affected by blockage of carotenoid synthesis. In the presence of diphenylamine, the B806 complex was formed in a much smaller amount than the RC-B870 complex. These results suggest that, in Erythrobacter sp. strain OCh114, bacteriochlorophyll plays an essential role in intracytoplasmic membrane development, and carotenoids are important for assembly of pigment-protein complexes.

Aerobiosis↗

Structural gene of cytochrome b-562 from the cytochrome b-c1 complex of Rhodobacter sphaeroides.

The structural gene coding for cytochrome b-562 isolated from the cytochrome b-c1 complex of Rhodobacter (Rhodopseudomonas) sphaeroides has been cloned. Its nucleotide sequence has been determined and the amino acid sequence was deduced therefrom. It consists of 157 amino acids (Mr 17,237) and contains four hydrophobic segments. The first 30 residues in the predicted amino acid sequence are the same as those determined for the NH2-terminal portion of purified cytochrome b-562. The amino acid composition is in accord with that determined for the pure protein. From the hydropathy profile and molar ratio of protoheme to cytochrome b-562, it is suggested that the structural and functional unit of the cytochrome is a two-heme cross-linked homodimer.

Amino Acid Sequence↗

A new membrane-bound b-type cytochrome, cytochrome b-558, from photosynthetically grown Rhodopseudomonas sphaeroides.

A new membrane-bound b-type cytochrome, cytochrome b-558, was removed from chromatophore membranes of photosynthetically grown Rhodopseudomonas sphaeroides strain R-26 by deoxycholate-cholate extraction. The cytochrome was purified by ammonium sulfate fractionation and ion-exchange chromatography. Cytochrome b-558 had absorption maxima at 280 and 405 nm in the oxidized form, and at 558, 528, and 420 nm in the reduced form. It had a midpoint potential of--130 mV at pH 7.0. The minimal molecular weight of this protein was 42,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it contained one mole heme per mole of protein. The isoelectric point was 8.5. The electrophoretic pattern of heme-carrying proteins and the redox potentiometry showed that cytochrome b-558 was present in membranes from wild type, strain R-26, and strain GA grown photosynthetically, but not from any strain grown aerobically.

Amino Acids↗

Characterization by EPR spectroscopy of cytochrome b-562 isolated from the cytochrome b-c1 complex of Rhodopseudomonas sphaeroides R-26.

The EPR spectra of cytochrome b-562 isolated from the cytochrome b-c1 complex of Rhodopseudomonas sphaeroides were measured at liquid helium temperature. The purified cytochrome b-562 gives a high spin signal at g = 6.0. Anaerobic titration of this signal confirmed the presence of two redox components with Em = 40 and -110 mV at pH 7.5. These values are consistent with the published ones, Em = 55 and -100 mV at pH 7.0, that were optically estimated for the same type of preparation (Iba et al. (1985) FEBS Lett. 183, 151-154). The power saturation behavior of the g = 6.0 signal at different redox potentials indicated a direct spin-spin interaction between these two redox centers.

Cytochrome b Group↗