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Biomedical subjects

I Yano

Publications and source records attributed to I Yano.

At least 163 records · Page 9Linked to original sources
PubMed1986

Mycobacterium fortuitum subspecies acetamidolyticum, a new subspecies of Mycobacterium fortuitum.

Mycobacterium fortuitum subspecies acetamidolyticum is a new subspecies of M. fortuitum and has an intermediate growth rate. It is a nonphotochromogenic mycobacterium. It does not utilize glutamate but utilizes acetamide as a simultaneous nitrogen and carbon source. It is able to utilize acetate, malate, pyruvate, fumarate, glucose, fructose, and n-propanol as the sole sources of carbon in the presence of ammoniacal nitrogen, but does not utilize them in the presence of glutamate-nitrogen. It is easily differentiated from all rapidly growing mycobacteria by its inability to utilize glutamate as a simultaneous nitrogen and carbon source, and from all slowly growing mycobacteria by its capacity to utilize acetamide as a simultaneous nitrogen and carbon source. Its mycolic acid pattern is different from that of M. fortuitum. However, its deoxyribonucleic acid showed 94% relatedness with that of M. fortuitum. In view of the above findings, it has been designated as a new subspecies of M. fortuitum. The organism was isolated from sputum of a 56-year-old patient with lung disease and is considered to be a lung pathogen. The type strain is ATCC 35931 (NCH E11620).

Base Composition↗
PubMed1986

Granuloma formation and hemopoiesis induced by C36-48-mycolic acid-containing glycolipids from Nocardia rubra.

As previously reported (I. Yano, I. Tomiyasu, S. Kitabatake, and K. Kaneda, Acta Leprologica 2:341-349, 1984), Nocardia rubra, one of the nonpathogenic actinomycetes, possesses three classes of mycolic acid-containing glycolipid, i.e., glucose mycolate, trehalose dimycolate, and trehalose monomycolate. The carbon chain length of their mycolic acids is shorter (C36-48) than that in mycobacteria (longer than C70), and the glycolipid consists of only alpha-mycolic acid. One intravenous administration of 500 micrograms of each purified glycolipid to ICR mice in the form of water-in-oil-in-water emulsion without any protein antigens caused prominent granuloma formation in the lungs, spleen, and liver. The lung index in the treated mice was about 3.5 times larger than that in the control mice (given water-in-oil-in-water emulsion only) at 1 week after the injection and then rapidly declined, while spleen and liver indices peaked at 2 weeks after the injection and persisted longer. The granuloma consisted of macrophages, some of which phagocytized glycolipid micelles, lymphocytes, monocytes, and neutrophils. In addition, many small hemopoietic islands were observed in the liver sinusoids, where various immature blood cells were trapped by the prominent cytoplasmic projections of Kupffer cells. The granuloma formation and hemopoiesis observed here are considered to be the most characteristic morphological expression of macrophage activation in these organs. This is the first report to show that such histological changes can be induced by chemically defined and homogeneous mycolic acid-containing glycolipids other than those of mycobacteria.

Animals↗
PubMed1986

Determination of molecular species composition of C80 or longer-chain alpha-mycolic acids in Mycobacterium spp. by gas chromatography-mass spectrometry and mass chromatography.

The molecular species composition of alpha-mycolic acids ranging from C68 to C86 in 13 rapidly growing and 12 slowly growing mycobacterial species was determined by gas chromatography, gas chromatography-mass spectrometry, and mass chromatography. In gas chromatographic analysis, the molecular species of alpha-mycolic acids were well separated as trimethylsilyl ether derivatives of the methyl esters, according to their total carbon numbers. The total carbon and double-bond numbers of mycolic acids at each peak on gas chromatograms were determined from the [M]+, [M - 15]+, and [M - 90]+ ions on the mass spectrum, and straight and branched chain structures were identified by the mass fragment ions [A]+, due to C2--C3 cleavage [R-CH-O-Si(CH3)3]+, and [B]+, due to C3--C4 cleavage [(CH3)3-Si-O-CH-CH(R')-COOCH3]+. The concentration of odd- and even-carbon-numbered mycolic acids, which often overlap each other on gas chromatograms, and the composition of three homologous mycolic acids with different alpha units (C22:0, C24:0, and C26:0) were clearly determined by mass chromatography monitoring [M - 15]+ ions and [B - 29]+ ions, respectively. The molecular species composition of alpha-mycolic acids and their average carbon numbers (av. cn.) as a simple expression of the composition were calculated from the mass chromatograms. Each mycobacterial species examined was demonstrated to possess a characteristic profile of alpha-mycolic acid composition, and based on this the species were classified approximately into eight groups: C68 to C76 (av. cn. 72), dienoic, possessing a C20 alkyl branch at the 2 position (C22 alpha-unit) for Mycobacterium diernhoferi and Mycobacterium sp. strain 3707, a chromogenic rapid grower; C72 to C78 (av. cn. 75), dienoic with both C22 and C24 alpha units, containing a small or a large amount of odd-carbon-numbered molecules, for M. vaccae, M. rhodesiae, and M. phlei (chromogenic rapid growers); C72 to C80 (av. cn. 75 to 77), dienoic with C24 alpha-unit, containing a moderate or a large amount of odd-carbon-numbered molecules, for M. smegmatis, M. chitae, M. chelonae (M. chelonei), and M. fortuitum (nonchromogenic rapid growers); C78 to C82 (av. cn. 80), even-carbon-numbered dienoic with C24 alpha unit for M. agri and M. thermoresistible (rapid growers); C75 to C81 (av. cn. 77 to 79), odd-carbon-numbered dienoic with C24 alpha unit for M. nonchromogenicum complex (M. nonchromogenicum, M. terrae, and "M. novum") (slow growers); (vi) C76 to C84 (av. cn. 79 to 81), even-carbon-numbered dienoic with C24 alpha unit for MAIS complex including M. scrofulaceum, M. avium, and M. intracellulare (slow growers); (vii) C72 to C80 (av. cn. 77 to 79), even-carbon-numbered dienoic with C24 alpha unit for M. szulgai, M. gordonae, and M. kansasii (chromogenic slow growers); and (viii) C76 to C86 (av. cn. 79 to 81), even-carbon-numbered dienoic with C26 alpha unit M. bovis Ravenol and BCG and M. tuberculosis H37Rv. This study demonstrated that gas chromatography-mass spectrometric analysis of the molecular species composition of alpha-mycolic acid can give rapid, important, and very precise information for the identification of pathogenic and nonpathogenic mycobacterial species.

Chromatography, Gas↗
PubMed1984

Separation and analysis of novel polyunsaturated mycolic acids from a psychrophilic, acid-fast bacterium, Gordona aurantiaca.

More than 30 molecular species of highly unsaturated mycolic acids, ranging from C60 to C78 and possessing between two and seven double bonds, have been obtained from a new genus of acid-fast bacteria, Gordona aurantiaca. They were fully separated and identified as their trimethylsilyl ether derivatives by a combination of silica gel thin-layer chromatography (TLC), argentation thin-layer chromatography and gas chromatography/mass spectrometry (GC/MS). On silica gel thin-layer chromatography two adjacent spots, corresponding to mycolic acids possessing different structures of straight-chain and alpha-alkyl branch, were detected. The lower spot was separated by argentation TLC into four subclasses: monoenoic (including a small amount of saturated), dienoic, trienoic and tetraenoic mycolic acids ranging from C62 to C74 and possessing a C16:0, C18:0 or C20:0 alkyl branch at the C-2 position. The upper spot was separated by argentation TLC into five subclasses: dienoic (including a small amount of monoenoic), trienoic, tetraenoic, pentaenoic and hexaenoic (heptaenoic) acids ranging from C64 to C78 and possessing a C18:1 or C20:1 alkyl branch at the C-2 position. These types of mycolic acid structure differ from those reported previously in Mycobacteria and Nocardia, in the numbers of both carbon atoms and double-bonds and the intermediate length of the alpha-alkyl branch. The characteristic polyenoic structure of the straight-chain alkyl unit was also confirmed by GC/MS analysis of the meromycolaldehydes obtained after pyrolysis of the methyl mycolates. The major aldehydes obtained from the lower-spot mycolic acids were C44, C46, C48, C50 and C52, while those from the upper-spot mycolic acids were C48, C50, C52, C54 and C56, centering at C54. These aldehydes were also shown to possess between two and four double bonds in the lower-spot and between two and seven double bonds in the higher-spot mycolic acids, respectively. The physiological role of such highly polyunsaturated mycolic acids in psychrophilic acid-fast bacteria is discussed.

Chemical Phenomena↗
PubMed1984

Isonicotinic acid hydrazide induced changes and inhibition in mycolic acid synthesis in Nocardia and related taxa.

The mycolic acid compositions of Nocardia rubra and related bacteria grown in media containing different concentrations of antituberculous isonicotinic acid hydrazide (INH) were determined in detail by gas chromatography-mass spectrometry. On the basis of molecular species composition, average carbon numbers of mycolic acids were calculated. In Nocardia rubra, N. lutea and Rhodococcus rhodochrous IFO-13161, the ratio of mycolic to non-mycolic fatty acids and the average carbon numbers of mycolic acids were decreased at the INH concentrations of higher than 1 microgram/ml, paralleling with the significant inhibition of growth. In above three species the synthesis of longer chain mycolic acids (longer than C44 or C46 ) was inhibited more significantly than shorter homologues such as C38 or C40 . In contrast, neither growth inhibition nor change in corynomycolic acid composition was observed in Corynebacteria xerosis and Rhodococcus rhodochrous IFO-13165 at the concentration region of INH up to 100 micrograms/ml. The direct mass fragmentographic analysis of the trimethylsilylated (TMS) derivatives of mycolic acid methyl esters, monitoring [M-15] ions of individual molecular species, revealed that the chain shortening of total mycolic acid molecule by INH occurred more greatly in more highly unsaturated subclasses than in less unsaturated subclasses. Furthermore, mass fragmentographic analysis, monitoring fragment ions (A) and (B), due to straight chain and branched chain alkyl units, respectively, demonstrated the inhibition of mycolic acids was not attributed to the shortening of alpha-alkyl chain, but to the inhibition of chain elongation of C28 to C32 straight chain meromycolic acids. It was also indicated the amounts of trehalose mono- and di- mycolate (cord factor) decreased significantly with the addition of INH (1 to 20 micrograms/ml) in the above strains. From the results obtained above, INH appeared to inhibit the synthesis of mycolic acids longer than C44 or C46 specifically by inhibiting chain elongation or desaturation of precursor long chain fatty acids longer than C28 or C30.

Actinomycetales↗
PubMed1984

Hydroxylation of prostaglandin A1 by the microsomes of rabbit intestinal mucosa.

Microsomes from rabbit small intestine mucosa were found to catalyze the hydroxylation of PGA1 in the presence of NADPH. The major product was identified as 20-hydroxy PGA1 by using high performance liquid chromatography and gas chromatography-mass spectrometry, and the minor product was assumed to be 19-hydroxy PGA1. The ratio of the former product to the latter was about 24.1. The specific PGA1 omega-hydroxylase activity of small intestine microsomes was comparable to that of liver microsomes, and was significantly higher than those of microsomes from other tissues such as kidney cortex and lung. Microsomes from rabbit colon mucosa also catalyzed the hydroxylation of PGA1 in the presence of NADPH, with the ratio of omega- to (omega-1)-hydroxy PGA1 formed being 33.0. The PGA1 hydroxylase activities of the microsomes from both small intestine and colon were inhibited markedly by carbon monoxide, indicating the participation of cytochrome P-450. A cytochrome P-450 was solubilized from small intestine microsomes, and purified to a specific content of 10.5 nmol of cytochrome P-450/mg of protein. This cytochrome hydroxylated PGA1 at the omega-position with a turnover rate of 38.2 nmol/min/nmol of cytochrome P-450 in the reconstituted system containing cytochrome P-450, NADPH-cytochrome P-450 reductase, cytochrome b5 and phosphatidylcholine. It is suggested that this cytochrome P-450 is specialized for the omega-hydroxylation of PGA1 in small intestine microsomes.

Animals↗
PubMed1984

Purification and characterization of cytochrome P-450 specific for prostaglandin and fatty acid hydroxylase activities from the microsomes of rabbit small intestinal mucosa.

Earlier studies (Kusunose, E., Kaku, M., Ichihara, K., Yamamoto, S., Yano, I., & Kusunose, M. (1984) J. Biochem. 95, 1733-1739) showed that a form of cytochrome P-450 isolated from microsomes of rabbit small intestinal mucosa had the highest prostaglandin A1 (PGA1) hydroxylase activity so far reported among cytochrome P-450s. The present paper describes the procedure for the purification and further characterization of this cytochrome (designated as cytochrome P-450ia). Cytochrome P-450ia had a monomeric molecular weight of 53,000. The CO-difference spectra of its reduced form showed a maximal absorption at 451 nm, and the absolute spectra of its oxidized form indicated that cytochrome P-450ia was present largely in the low-spin state, and partially in the high-spin state. The cytochrome efficiently catalyzed the hydroxylation of fatty acids as well as prostaglandins in a reconstituted system containing cytochrome P-450, NADPH-cytochrome P-450 reductase, phospholipid, and cytochrome b5. PGA1 was the most efficient substrate, followed by myristate, laurate, palmitate, caprate, and PGE1 or PGE2. Among phospholipids, didecanoyl- and dilauroylphosphatidylcholines had the most stimulatory effect for both activities. 20-Hydroxy PGA1 was identified as the hydroxylation product of PGA1 by gas chromatography-mass spectrometry and mass fragmentography; the possibility of 19-hydroxy PGA1 being the product was excluded. In contrast, both omega- and (omega-1)-hydroxy fatty acids were identified as hydroxylation products of fatty acids. Cytochrome P-450ia had no detectable activity toward aminopyrine, benzphetamine, p-nitroanisole, 7-ethoxycoumarin, benzo(a)pyrene, or hexadecane.

Animals↗
PubMed1983

Ornithine-containing lipid of Bordetella pertussis, a new type of hemagglutinin.

The ornithine-containing lipids of six strains (phases I-IV) of Bordetella pertussis were prepared from the total extractable cellular lipids by thin-layer chromatography and treatment with phospholipase A. They were compared with those prepared from two strains each of Bordetella parapertussis and Bordetella bronchiseptica. The structures of the ornithine-containing lipid of B. pertussis and the other two species were resolved by acid and alkaline hydrolysis, gas-liquid chromatography, infrared absorption spectroscopy, amino acid analysis and combined gas-liquid chromatography/mass spectrometry. The main structure of the aminolipid of the three species of Bordetella was 3-hydroxyhexadecanoic acid, amide-linked to ornithine and esterified to the second hexadecanoic acid. The aminolipid of B. pertussis Sakurayashiki (phase III) exhibited high hemagglutinating activity for human and rabbit erythrocytes, having a minimum hemagglutinating concentration of 1 microgram/ml against 8-16 micrograms/ml for the other strains of Bordetella. All of these aminolipids showed some degree of microheterogeneity. Because the 3-hydroxyhexadecanoic acid content was especially high in strain Sakurayashiki, it was presumed that the intensity of hemagglutinating activity of the aminolipid was affected by the chain length of the central 3-hydroxy fatty acid, that is the aminolipid containing 3-hydroxyhexadecanoic acid had high hemagglutinating activity. The hemagglutination was inhibited by phosphatidylcholine at concentrations of more than 20 micrograms/ml. Other inhibitory substances were cysteine, sphingomyelin, acidic amino acids, histidine, unsaturated fatty acid and basic amino acids. Furthermore, the divalent cations Ca2+ and Mg2+ inhibited this hemagglutination at a concentration of 1 mM. The O-deacylated ornithine-containing lipid that had lost hexadecanoic acid did not have any hemagglutinating activity but did have hemolytic activity. Observation by electron microscopy indicated that erythrocytes were combined by the liposomes of the ornithine-containing lipids. On the basis of these results, the proposed mechanism of hemagglutination by the aminolipids is that the liposomes of the aminolipids combine erythrocytes by hydrophobic interaction between the fatty acid moieties of the aminolipid and the lipids of the surface of erythrocytes, and by ionic interaction between the ornithine of the aminolipid and the protein of the surface of the erythrocytes. In addition, the hemagglutinating activity of phosphatidylserine was found to be due to its similar structure to that of the ornithine-containing lipid and the mechanism was also presumed to be similar. The mechanism of hemagglutination by these aminolipids was distinct from that of lectins.

Bordetella↗
PubMed1980

ESR studies on the membrane properties of a moderately halophilic bacterium. I. Physical properties of lipid bilayers in whole cells.

The lipid membrane properties of a moderately halophilic gram-negative bacterium, Pseudomonas halosaccharolytica ATCC 29423, were studied by the use of stearate spin labels, 5NS, 12NS, and 16NS, changing the temperature of ESR measurement from 15 to 50 degrees C. The order parameter and the rotational correlation time of the spin labels incorporated into intact cell membranes of this bacterium grown at various temperatures in media containing different NaCl concentrations were calculated. The activation energy of rotational microviscosity was obtained from Andrade plots. At low growth temperature and low NaCl concentration in the medium, extractable lipids of this bacterium contained comparatively large amounts of unsaturated fatty acids, but as the growth temperature and NaCl concentration in the medium increased, the contents of saturated and cyclopropanoic fatty acids increased to more than half of the total fatty acids. 5NS gave the highest order parameters for the intact cells of this bacterium, while 12NS gave lower and 16NS gave the lowest results. The order parameters of 5NS, 12NS, and 16NS were completely separated, and all order parameters decreased gradually as the measuring temperature was increased. In contrast, the rotational correlation times of the intact cells with 12NS were as large as those with 5NS, while those with 16NS were distinctly smaller. Increasing NaCl concentrations in the growth medium caused an increase of the rotational correlation times, that is, stiffened the lipid bilayers. The Andrade plot for 16NS was approximately a straight line, whereas 5NS and 12NS gave two straight lines crossing at a temperature near the growth temperature, indicating phase transition from solid to liquid. The microviscosity activation energies were 5--10 kcal/mol in the liquid phase and 15--25 kcal/mol in the solid phase.

Cell Membrane↗