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Biomedical subjects

M Fujioka

Publications and source records attributed to M Fujioka.

At least 181 records · Page 10Linked to original sources

[A case of pulmonary aspergillosis presenting pulmonary bulla and eosinophilic pneumonia].

A rare case of a 25-year-old man with pulmonary aspergillosis is reported. Pulmonary bulla and eosinophilic pneumonia in the right upper lobe were diagnosed by chest roentgenogram and transbronchial lung biopsy. Because the patient developed infective bullae during steroid therapy, we performed transcutaneous thoracic drainage and right upper lobectomy. The resected lung tissue contained numerous hyphae of Aspergillus. Around the hyphae of Aspergillus, granulomatous reaction and eosinophilic infiltration were observed. Antibodies against Aspergillus were detected in the serum of the patient. The number of peripheral blood eosinophils decreased after right upper lobectomy. These findings suggest that pulmonary bullae and eosinophilic pneumonia may be a rare manifestation of pulmonary aspergillosis.

Adult↗

Low-energy collision-induced dissociation of protonated polyamino alcohol derivatives of peptides and N-terminal blocked peptides.

This investigation reports the low-energy collision-induced dissociation of the protonated molecules of polyamino alcohols, formed by chemical reduction of synthetic peptides and N-terminal blocked peptides, in order to evaluate its potential for peptide sequence determination. The --CH2--NH-- cleavage with charge retention on the N-terminus was prominent, and the entire sequence of ions produced in this manner was observed. Some of the sequence ions arising from --CH2--NH-- cleavage accompanied by migration of two hydrogens, and --NH--C alpha H-- cleavage with charge retention on the C-terminus, also occurred prominently. In addition, a great number of internal fragment ions were produced in relatively high abundance; these provided supplementary sequence information and were, in some cases, critical in determining the sequence. The usefulness of this method was exemplified by its application to the sequence determination of bacterial lipopeptides.

Lipoproteins↗

Site-directed mutagenesis of rat liver S-adenosylhomocysteinase. Effect of conversion of aspartic acid 244 to glutamic acid on coenzyme binding.

Aspartic acid 244 that occurs at the putative NAD(+)-binding site of rat liver S-adenosylhomocysteinase was replaced by glutamic acid by oligonucleotide-directed mutagenesis. The mutant enzyme was purified to homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel permeation chromatography showed that the purified mutant enzyme was a tetramer as is the wild-type enzyme. In contrast to the wild-type enzyme, which possesses 1 mol of tightly bound NAD+ per mol of enzyme subunit, the mutant enzyme had only 0.05 mol of NAD+ but contained about 0.6 mol each of NADH and adenine per mol of subunit. The mutant enzyme, after removal of the bound compounds by acid-ammonium sulfate treatment, exhibited S-adenosylhomocysteinase activity when assayed in the presence of NAD+. From the appearance of activity as a function of NAD+ concentration, the enzyme was shown to bind NAD+ with a Kd of 23.0 microM at 25 degrees C, a value greater than 280-fold greater than that of the wild-type enzyme. In the presence of a saturating concentration of NAD+, the mutant enzyme showed apparent Km values for substrates similar to those of the wild-type enzyme. Moderate decreases of 8- and 15-fold were observed in Vmax values for the synthetic and hydrolytic directions, respectively. These results indicate the importance of Asp-244 in binding NAD+, and are consistent with the idea that the region of S-adenosylhomocysteinase from residues 213 to 244 is part of the NAD+ binding site. This region has structural features characteristic of the dinucleotide-binding domains of NAD(+)- and FAD-binding proteins (Ogawa, H., Gomi, T., Mueckler, M. M., Fujioka, M., Backlund, P.S., Jr., Aksamit, R.R., Unson, C.G., and Cantoni, G.L. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 719-723).

Adenosylhomocysteinase↗

Rat serine dehydratase gene codes for two species of mRNA of which only one is translated into serine dehydratase.

With the previously obtained rat liver serine dehydratase cDNA (SDH2; Ogawa, H., Miller, D.A., Dunn, T., Su, Y., Burcham, J. M., Peraino, C., Fujioka M., Babcock, K., and Pitot, H. C. (1988) Proc. Natl. Acad. Sci. U.S. A. 85, 5809-5813) as a probe, we isolated a different species of cDNA (SDH3) from the same cDNA library from which SDH2 was obtained. Nucleotide sequence analysis has indicated that SDH3 has an open reading frame which encodes 327 amino acid residues and which is identical to that of the cDNA obtained by Noda et al. (Noda, C., Ito, K., Nakamura, T., and Ichihara, A., (1988) FEBS Lett. 234, 331-335). Primer extension analysis and RNase protection mapping clarified that the SDH3 mRNA was the major mRNA for serine dehydratase in the liver, and its transcription begins with a T residue located 23 nucleotides down-stream of a TATA-like box. In vitro transcription/translation experiment demonstrated that SDH3 encoded a polypeptide of 35 kDa, a size in agreement with that of the subunit of the purified protein, whereas SDH2, despite having a size larger than SDH3, produced a peptide of much smaller size that reacted with anti-serine dehydratase IgG. SDH2 was found to have a stop codon early in the sequence and is predicted to encode a polypeptide of 8.9 kDa. Also, SDH2 has a 5'-noncoding sequence different from that of SDH3. These results indicate that alternative transcription initiation and different modes of splicing of the primary transcripts of rat serine dehydratase gene result in the formation of two species of mRNA, of which only one is translated into the mature serine dehydratase protein.

Amino Acid Sequence↗

Diagnostic values of gallbladder hyperperfusion and the rim sign in radionuclide angiography and hepatobiliary imaging.

The diagnostic values of hyperperfusion to the gallbladder fossa and the rim sign were retrospectively evaluated in 24 histopathologically proven cases of gallbladder disease. Although 12 of 13 patients (92%) with acute cholecystitis had nonvisualization of the gallbladder, the positive predictive value with this finding was 71%. However, when the finding was associated with hyperperfusion to the gallbladder fossa or the rim sign, the positive predictive value with both (90%) was markedly increased, compared to the prevalence ratio of 54%. Such a combination could be useful for preserving a high positive predictive value in the diagnosis of acute cholecystitis.

Acute Disease↗

[Dental findings in a case of Chondrodysplasia Punctata].

Chondrodysplasia Punctata is a disease belonging to the metaphysia syndrome. This disease is characterized by endochondral punctata calcification of epiphysis or apophysis, short legs and hands, arthrogryposis, saddle nose, ichthyosis, cataract and mental retardation. This report concerns a case of Chondrodysplasia Punctata (Rhizomeric form) in a 4 year 1 month old girl. Observations were made from the dental point of view, and the findings were as follows: 1) The mesio-distal and bucco-lingual lengths of all primary teeth were smaller than the standard size. 2) Measuring the dental cast, the width of the dental arch was found to be larger, but the length of the arch was found to be smaller than the standard size. 3) The value of the palatal height was slightly smaller, compared with that of the normal children. 4) According to X-ray cephalometric analysis, the growth of the mandible was insufficient. 5) An enlargement of the angles of the teeth axes of the upper and lower central incisors was found. 6) The growth of the basal arches of the maxilla and mandible were shown to be retarded in the anterior portion. 7) Measurement of the area of the tubella sella triangle, revealed the posterior parts to be large and almost standard in the mesofacial part, and small in the mandibulo-facial parts.

Child, Preschool↗

[Present aspect of infection and incidence of tuberculosis in youth registered in Aichi Prefecture].

In this study, 151 school children and students with tuberculosis registered in Aichi Prefecture during 1983 to 1987 were investigated. The results were as follows: 1) The extent of lesions were not so advanced, as shown by the fact that 27.2% were cavitary and 31.8% were bacteriologically confirmed. 2) They were detected mainly by symptomatic visit to physicians (39.1%), periodic mass radiography (31.8%) and contacts survey (20.6%). The proportion of the former symptomatic visit was comparatively lower than in other age groups. 3) Patient's delay in cases detected by symptomatic visit was not so evident, however, there is a certain grade of doctor's delay; for instance, 22.0% of them took 2 months and more to be diagnosed. 4) 77.5% of them were vaccinated with BCG once or more, in most cases more than 5 years before their detection. 5) Sources of infection were specified in 52.3% of the cases. Most of them were found in their household and were bacteriologically confirmed at the time of detection. In some students their classmates were suspected as their possible source of infections.

Adolescent↗

The neutrophil chemoattractant produced by the rat kidney epithelioid cell line NRK-52E is a protein related to the KC/gro protein.

A basic protein having chemotactic activity for neutrophils is secreted by the rat kidney epithelioid cell line NRK-52E in response to interleukin-1 beta (Watanabe, K., Kinoshita, S., and Nakagawa, H. (1989) Biochem. Biophys. Res. Commun. 161, 1093-1099). The protein, which is referred to as cytokine-induced neutrophil chemoattractant (CINC), has been shown to be a dimer of identical subunits; and the complete amino acid sequence of the subunit has been established. Sequence determination has been achieved by automated Edman degradation of reduced and carboxymethylated CINC and of peptides generated by cleavage with cyanogen bromide and lysyl endopeptidase. The CINC subunit consists of 72 amino acid residues. The amino acid sequence of CINC shows striking similarities to the sequences of the proteins encoded by the mouse platelet-derived growth factor-inducible KC gene and human and hamster gro genes, suggesting that CINC is the rat counterpart of the KC/gro protein.

Amino Acid Sequence↗

Human liver serine dehydratase. cDNA cloning and sequence homology with hydroxyamino acid dehydratases from other sources.

Rat liver serine dehydratase cDNA was used to screen a human liver cDNA library in lambda gt11. One positive clone occurred in every 5,000 clones. Fifteen positive clones were plaque purified. The largest cDNA obtained contained an open reading frame of 987 base pairs, and 5' and 3' noncoding regions of 89 and 317 base pairs, respectively. The deduced amino acid sequence, with a calculated Mr of 34,615, was similar to that of rat liver serine dehydratase except for the absence of a segment consisting of 36 amino acid residues. In vitro transcription/translation with the cDNA resulted in the formation of a polypeptide with an Mr of approximately 35,000, which cross-reacted with the anti-rat serine dehydratase antibody. These results suggest that the human serine dehydratase is structurally cognate with the rat enzyme. Moreover, portions of the sequence postulated to be essential for activity in microbial threonine dehydratases are found in the mammalian serine dehydratases, suggesting that hydroxyamino and dehydratases may have originated from a common ancestor.

Amino Acid Sequence↗

Expression of rat liver S-adenosylhomocysteinase cDNA in Escherichia coli and mutagenesis at the putative NAD binding site.

The cDNA for rat liver S-adenosylhomocysteinase has been cloned, and the nucleic acid sequence has been determined. By comparison of the deduced amino acid sequence for S-adenosylhomocysteinase with that of the dinucleotide binding region for other proteins, the sequence from amino acids 213 to 244 in rat liver S-adenosylhomocysteinase was proposed to be part of the NAD binding site (Ogawa, H., Gomi, T., Mueckler, M. M., Fujioka, M., Backlund, P. S., Jr., Aksamit, R. R., Unson, C. G., and Cantoni, G. L. (1987) Proc. Natl. Acad. Sci. U. S. A. 84, 719-723). A vector has been constructed that expresses S-adenosylhomocysteinase in Escherichia coli in the presence of isopropyl beta-D-thiogalactopyranoside by inserting the coding sequence of rat liver S-adenosylhomocysteinase cDNA downstream from the lac promoter of plasmid pUC118. The enzyme that is produced comprises as much as 10% of the soluble cellular proteins. The purified enzyme is a tetramer, contains 4 mol of tightly bound NAD, and has kinetic properties indistinguishable from those of the liver enzyme. Tryptic peptide mapping and NH2-terminal sequence analysis indicate that the recombinant enzyme is structurally identical to the liver enzyme except for the absence of the NH2-terminal blocking group. The rat liver enzyme has a blocked NH2-terminal alanine residue (Ogawa, H., Gomi, T., Mueckler, M. M., Fujioka, M., Backlund, P. S., Jr., Aksamit, R. R., Unson, C. G., and Cantoni, G. L. (1987) Proc. Natl. Acad. Sci. U. S. A. 84, 719-723). By oligonucleotide-directed mutagenesis mutant vectors have been generated that express proteins in which each of the glycines in the Gly-Xaa-Gly-Xaa-Xaa-Gly sequence of the putative NAD binding site (residues 219-224) was changed to valine. Immunoblot analysis of extracts of the cells transformed with these vectors reveals the presence of immunoreactive proteins with the subunit molecular weight of S-adenosylhomocysteinase. The mutant proteins have no catalytic activity, contain no bound NAD, and do not form the same quaternary structure as the recombinant S-adenosylhomocysteinase.

Adenosylhomocysteinase↗

The peptide sequences near the bound pyridoxal phosphate are conserved in serine dehydratase from rat liver, and threonine dehydratases from yeast and Escherichia coli.

The blocked amino-terminal residue of rat liver serine dehydratase was shown to be acetylalanine by analysis of an isolated amino-terminal peptide after digestion with acylamino acid-releasing enzyme. Digestion of the borohydride-reduced, carboxymethylated enzyme with lysyl endopeptidase yielded a single epsilon-N-pyridoxyllysine-containing peptide, whose sequence is Met-Asp-Ser-Ser-Gln-Pro-Ser-Gly-Ser-Phe-Lys(Pxy)-Ile-Arg-Gly- His-Leu-Cys(Cm)-Lys. This peptide comprises residues 30-49 of the cDNA-deduced amino acid sequence. The sequence of seven amino acids around the bound pyridoxal phosphate is highly conserved in serine dehydratase from rat liver, and threonine dehydratases from yeast and Escherichia coli.

Amino Acid Sequence↗

Bunazosin, an alpha 1-adrenoceptor blocker, differentially releases co-transmitters in dog mesenteric vessels.

The effects of bunazosin on the electrical and mechanical responses of smooth muscle cells elicited by exogenously applied noradrenaline (NA) and by perivascular nerve stimulation were studied in the isolated mesenteric artery and vein of the dog. NA (above 10(-7) M in the artery and above 3 X 10(-8) M in the vein) depolarized the membrane. Perivascular nerve stimulation evoked an excitatory junction potential (e.j.p.) and slow depolarization in both vessels. Bunazosin and prazosin inhibited the NA-induced depolarization and slow depolarization in the artery but not in the vein. The NA actions in the vein were inhibited by yohimbine. Bunazosin (above 10(-6) M) increased the amplitude of the e.j.p. but decreased the outflow of NA during nerve stimulation. The amplitude and conduction velocity of the compound action potential of perivascular nerves were inhibited by higher concentrations of bunazosin (above 10(-5) M). The results provide evidence that bunazosin has selective inhibitory actions at alpha 1-adrenoceptors. This drug exerted differential effects on the release of co-transmitters which generate the e.j.p. and the slow depolarization, as bunazosin increased the former and decreased the latter. This suggests that e.j.p. is generated by a substance other than NA.

Action Potentials↗

Rat liver S-adenosylhomocysteinase. Spectrophotometric study of coenzyme binding.

Rat liver S-adenosylhomocysteinase, a homotetramer, was resolved by treatment with acid ammonium sulfate into apoenzyme and NAD. The apoenzyme thus prepared retained a tetrameric structure but differed in the mobility on nondenaturing polyacrylamide gel electrophoresis. The inactive apoenzyme was reactivated upon incubation with NAD. The restoration of activity paralleled with the tight binding of NAD to apoenzyme, and full activity was obtained when 4 mol of NAD were bound per mol of apoenzyme. The kinetics of reconstitution were apparently biphasic and suggest the existence of two conformers in a slow equilibrium, one of which binds the coenzyme rapidly while the other does so very slowly, if at all. In addition to NAD, apoadenosylhomocysteinase tightly bound nicotinamide hypoxanthine dinucleotide, 3-acetylpyridine adenine dinucleotide and nicotinic acid-adenine dinucleotide. NADP was not bound. Catalytic activity was found only with the enzyme reconstituted with NAD or nicotinamide hypoxanthine dinucleotide. The spectral change observed on interaction of apoadenosylhomocysteinase with NAD was similar to those seen with adenine nucleotides, and was largely approximated by the addition of dioxane to aqueous solutions of adenine nucleotides. By comparison of the difference spectra, it is suggested that the adenine portion of the coenzyme is bound in the hydrophobic pocket of the protein, and that the binding is accompanied by perturbation of tryptophan residue of the protein.

Adenosylhomocysteinase↗

Rational use of diagnostic imaging in paediatrics. The report of a World Health Organization Study Group.

The World Health Organization (WHO) is the agency of the United Nations with primary responsibility for international health matters and public health. Through the exchange of the knowledge and experience of health professionals from 165 nations WHO promotes comprehensive health services worldwide. As a part of this effort WHO has in recent years been concerned with improving the availability and utilization of diagnostic imaging, especially as it relates to health services in developing countries. Having devised and specified suitable basic radiographic equipment (the WHO BRS) a WHO Scientific Group was convened in 1982 to study the utilization of radiographic procedures and to to recommend their more efficacious and rational use. Then in 1984, recognizing the implications of newer imaging technology on world health, a second WHO Scientific Group considered the design, impact and future use of ultrasonography and computed tomography in developing countries. The findings of these scientific groups were published as part of the WHO Technical Report Series which makes available the recommendations of such international groups of experts. Although some of the needs of children were considered in these previous Technical Reports, WHO felt that the size and socioeconomic importance of the paediatric population worldwide warranted a further study and report to address the particular technical and clinical problems of diagnostic imaging in the paediatric age group. Thus a third WHO Scientific Group met in Geneva from 18 to 24 November 1985 to review established practices and experience in paediatric imaging throughout the world; and to recommend rational guidelines for diagnostic imaging for children to age 14 years.(ABSTRACT TRUNCATED AT 250 WORDS)

Child↗

Purification of an autocrine growth factor in conditioned medium obtained from primary cultures of scleral fibroblasts of the chick embryo.

Scleral fibroblasts of the chick embryo were found to secrete autocrine growth factors. One of the factors was purified from conditioned medium collected from growing-phase cultures of these cells by DEAE-Sepharose column chromatography and following non-denaturing polyacrylamide gel electrophoresis. The specific activity was increased 1100-fold by this purification. The chromatographically purified growth factor was still active after incubation at 95 degrees C, at pH 10 or pH 3, or with glycosidase H, but inactive after incubation with dithiothreitol or trypsin. An active protein having a molecular weight of 32 kDa was found to be the major component of the final preparation.

Animals↗

Scleral fibroblasts of the chick embryo proliferate by an autocrine mechanism in protein-free primary cultures: differential secretion of growth factors depending on the growth state.

Scleral fibroblasts of the chick embryo in primary culture proliferated in a protein-free medium. Conditioned medium (CdM) from the culture contained plural growth-promoting factors, which were active to the same cell type. The activity of one of the growth-promoting factors (SAF-I) was heat-resistant and the rest (SAF-II) were heat-sensitive. SAF-I accumulated in the CdM only during the growing phase; on the other hand, SAF-II accumulated in the CdM during the stationary phase. SAF-I showed the same time course of DNA synthesis-promoting activity as human PDGF. However, the activity of the SAF-I was not neutralized by anti-human PDGF. On the other hand, a part of the SAF-II (SAF-II a) showed a strong affinity for heparin.

Animals↗

Endogenous and exogenous testosterone levels after administration of deuterium-labelled testosterone propionate in hypogonadotropic hypogonadism.

A gas chromatography-mass spectrometry-selected ion monitoring technique was employed to determine simultaneously the plasma concentrations of endogenous and exogenous testosterone in three patients with hypogonadotropic hypogonadism after a single i.m. dose of testosterone propionate-19,19,19-d3. The plasma levels of testosterone-19,19,19-d3 derived from testosterone propionate-19,19,19-d3 were maintained above the normal testosterone levels (greater than 4 ng/ml) for 48 h, while the plasma levels of endogenous testosterone changed little.

Adult↗