PubMed Health⌕ Search

Biomedical subjects

S Takaichi

Publications and source records attributed to S Takaichi.

At least 37 records · Page 2Linked to original sources
PubMed1997

New carotenoids from the thermophilic green sulfur bacterium Chlorobium tepidum: 1',2'-dihydro-gamma-carotene, 1',2'-dihydrochlorobactene, and OH-chlorobactene glucoside ester, and the carotenoid composition of different strains.

The complete carotenoid composition of the thermophilic green sulfur bacterium Chlorobium tepidum strain TNO was determined by spectroscopic methods. Major carotenoids were four kinds of carotenes: gamma-carotene, chlorobactene, and their 1',2'-dihydro derivatives (1',2'-dihydro-gamma-carotene and 1',2'-dihydrochlorobactene). In lesser amounts, hydroxyl gamma-carotene, hydroxyl chlorobactene, and their glucoside fatty acid esters were found. The only esterified fatty acid present was laurate, and OH-chlorobactene glucoside laurate is a novel carotenoid. In other strains of C. tepidum, the same carotenoids were found, but the composition varied from strain to strain. The overall pigment composition in cells of strain TNO was 4 mol carotenoids and 40 mol bacteriochlorophyll c per mol bacteriochlorophyll a. The effects of nicotine on carotenoid biosynthesis in C. tepidum differed from those in the thermophilic green nonsulfur bacterium Chloroflexus aurantiacus.

Carotenoids↗
PubMed1997

The major carotenoid in all known species of heliobacteria is the C30 carotenoid 4,4'-diaponeurosporene, not neurosporene.

The carotenoids of five species of heliobacteria (Heliobacillus mobilis, Heliophilum fasciatum, Heliobacterium chlorum, Heliobacterium modesticaldum, and Heliobacterium gestii) were examined by spectroscopic methods, and the C30 carotene 4,4'-diaponeurosporene was found to be the dominant pigment; heliobacteria were previously thought to contain the C40 carotenoid neurosporene. In addition, trace amounts of the C30 diapocarotenes diapolycopene, diapo-zeta-carotene, diapophytofluene, and diapophytoene were also found. Up to now, diapocarotenes have been found in only three species of chemoorganotrophic bacteria, but not in phototropic organisms. Furthermore, the esterifying alcohol of bacteriochlorophyll g from all known species of heliobacteria was determined to be farnesol (C15) instead of the usual phytol (C20). Heliobacteria may be unable to produce geranylgeranyol (C20).

Bacteria↗
PubMed1997

Globular and fibrous structure in barley chromosomes revealed by high-resolution scanning electron microscopy.

Barley chromosomes were prepared for high-resolution scanning electron microscopy using a combination of enzyme maceration, treatment in acetic acid and osmium impregnation using thiocarbohydrazide. Using this technique, the three-dimensional ultrastructure of interphase nuclei and mitotic chromosomes was examined. In Interphase, different levels of chromatin condensation were observed, consisting of fibrils 10 nm in diameter, 20- to 40-nm fibres and a higher order complex. In prophase, globular and strand-like structures composed of 20- to 40-nm fibres were dominant. As the cells progressed through the cell cycle and the chromatin condensed, globular and strand-like structures (chromomeres) were coiled and packed to form chromosomes. Chromomeres were observed as globular protuberances on the surface of metaphase chromosomes. These findings indicate that the chromomere is a fundamental substructure of the higher order architecture of the chromosome. In the centromeric region, there were no globular protuberances, but 20- to 40-nm fibres were folded compactly to form a higher level organization surrounding the chromosomal axia.

Chromatin↗
PubMed1997

Porphyrobacter tepidarius sp. nov., a moderately thermophilic aerobic photosynthetic bacterium isolated from a hot spring.

A new thermophilic bacterium, strain OT3T (T = type strain), was isolated from a brackish hot spring. Strain OT3T is an obligate aerobe that synthesizes bacteriochlorophyll a and has a photosynthetic apparatus. This isolate is a thermophilic bacterium with an optimal growth temperature of 40 to 48 degrees C. The cells are nonmotile, ovoid to short rods. An analysis of 16S rRNA sequences revealed that the new strain forms a coherent cluster with members of the alpha-4 group of the alpha subclass of the Proteobacteria, which contains the genera Erythrobacter, Erythromicrobium, and Porphyrobacter. The closest relative is Porphyrobacter neustonensis, with a 16S rRNA sequence similarity of 96.8%. The in vivo absorption spectrum has maxima at 460, 494, 596, 800, and 870 nm. The main carotenoids are OH-beta-carotene sulfate derivatives, nostoxanthin, and bacteriorubixanthinal. Growth occurs with glucose, acetate, glutamate, butyrate, Casamino Acids, and yeast extract as sole energy sources. The pigment composition and nutritional profile of the new isolate are similar to the pigment composition and nutritional profile of P. neustonensis. Although there are marked differences in cell morphology between the new isolate and the budding bacterium P. neustonensis, the results of phenotypic and genetic comparisons suggest that the new isolate is closely related to P. neustonensis. Consequently, we assign the new isolate to the genus Porphyrobacter and propose the name Porphyrobacter tepidarius sp. nov. for it; the type strain of P. tepidarius is strain OT3 (= DSM 10595).

Bacteria, Aerobic↗
PubMed1996

The carotenoid 7,8-dihydro-psi end group can be cyclized by the lycopene cyclases from the bacterium Erwinia uredovora and the higher plant Capsicum annuum.

The genes for geranylgeranyl diphosphate synthase (crtE) and phytoene synthase (crtB) from the epiphytic bacterium Erwinia uredovora and the phytoene desaturase gene from the photosynthetic bacterium Rhodobacter capsulatus (Rc-crtI) were introduced into Escherichia coli, which resulted in the accumulation of the acyclic carotenoid, neurosporene. Further introduction of the lycopene cyclase gene from E. uredovora (crtY) or the higher plant Capsicum annuum (Icy) resulted in the production of a bicyclic carotenoid, 7,8-dihydro-beta-carotene, via monocyclic beta-zeacarotene. zeta-Carotene was also found to be cyclized to bicyclic 7,8,7',8'-tetrahydro-beta-carotene by the Erwinia cyclase. These results indicate that both lycopene cyclases can cyclize a 7,8-dihydro-psi end group to a 7,8-dihydro-beta end group, in addition to the usual cyclization of the psi end group to the beta end group. Furthermore, beta-carotene hydroxylase from Erwinia (CrtZ) was able to add a hydroxyl group to the 7,8-dihydro-beta end group and the beta end group.

Bacterial Proteins↗
PubMed1996

Epithelial-mesenchymal transformation of a newly established cell line from ovarian adenosarcoma by transforming growth factor-beta1.

We report on 2 cell lines (designated OVAK-I and OVAK-II) established from an adenosarcoma, a rare variant of a malignant Mullerian mixed tumor, of the ovary. OVAK-I was not tumorigenic in nude mice and showed no responsiveness to transforming growth factor (TGF)-beta1, whereas OVAK-II was tumorigenic and had various biological properties. Light- and electron-microscopic studies showed that the morphology of the cells was converted from an epithelial to a mesenchymal form by TGF-betaI. Furthermore, the protein and mRNA expressions of the epithelial marker carcinoembryonic antigen and the mesenchymal marker vimentin were down- and up-regulated, respectively, by TGF-beta1. These findings suggested that TGF-beta1 plays a role in epithelial-mesenchymal transformation and support the single-stem-cell theory which explains the origin of malignant Mullerian mixed tumors: that the epithelial and mesenchymal components of these tumors are of common stem-cell origin.

Adenosarcoma↗
PubMed1996

Histological changes in brain tissue and vasculature after intracarotid infusion of organic solvents in rats.

Organic solvents, such as ethanol or dimethyl sulphoxide (DMSO), have been used in liquid embolic agents. To investigate the effects of these solvents on the cerebral blood vessels and cerebral tissue, we subjected Wistar rats weighing 250-300 g to internal carotid artery infusion of 0.2 ml diluted ethanol (10%, 40% or 70%) or anhydrous DMSO (100%). Some rats were sacrificed 5 min after the infusion and the remainder at 10 days. Rats injected with ethanol at high concentration or DMSO showed extensive exudation of Evans blue at the site of injection 5 min after infusion, together with full-thickness necrosis of the wall of vessels and swelling of brain cells. In contrast, rats injected with 10% or 40% ethanol solution showed necrosis of only the intimal layer and partial necrosis of the medial layer and no brain swelling was observed. These findings suggest that ethanol at low concentration can be used as a relatively safe solvent for liquid embolic substances.

Animals↗
PubMed1996

Influence of prolonged ventricular assistance on myocardial histopathology in intact heart.

BACKGROUND: The unloading effect of ventricular assistance on the injured myocardium may adversely affect the compensatory hypertrophy of the residual intact myocardium because myocardial protein synthesis is partly controlled by cardiac work. The influence of prolonged ventricular assistance on normal myocardium was evaluated from a pathologic point of view. METHODS: A ventricular assist device was chronically implanted in 5 goats using left atrium-aorta bypass. The pumping ratio was fixed at 70 beats/min. Left ventricular biopsy samples were taken before and 1 month after assistance. RESULTS: Although the volume densities of myocytes and interstitial tissue in the myocardium showed no significant changes after 1 month of support, the myocyte volume density to nuclear volume density ratio and the interstitial tissue volume density to nuclear volume density ratio decreased significantly (p < 0.01 and p < 0.05, respectively). A cross-sectional area of myocyte showed decreases of 20.9% to 49.5%, whereas the nuclear cross-sectional area showed no significant changes. In addition, myofibrillar volume density in the cytoplasm decreased from 54.9 +/- 2.3% to 49.1 +/- 4.4%. CONCLUSIONS: The results indicate that long-term ventricular assistance in the intact heart leads to myocardial atrophy. This suggests that in the damaged heart subjected to prolonged unloading by ventricular assistance, there is the possibility of limiting compensatory hypertrophic changes in the residual intact myocardium.

Animals↗
PubMed1995

Ultrastructural localization and translocation of nitric oxide synthase in the endothelium of the human cerebral artery.

An electron microscopic immunocytochemical study was undertaken to clarify ultrastructural localization and translocation of nitric oxide synthase (NOS) in endothelial cells (EC) of the human cerebral and superficial temporal arteries (STA) employing antibody against endothelial NOS (EC-NOS). NOS immunoreactivity was found in all EC examined, in association with the plasma membrane and cytoplasmic organelles such as endoplasmic reticulum, Weibel-Palade body and subplasmalemmal vesicles, and in the cytoplasm devoid of organelles and extracellular regions, irrespective of arteries. The immunoreactivity in subplasmalemmal vesicles was, however, demonstrated only in human cerebral arteries. In the human STA exposed to bradykinin which induces EC-NOS phosphorylation, the gold particles significantly increased in the cytosol and decreased in the areas associated with cytoplasmic organelles; however, the number of particles did not change significantly in the plasma membrane. The results implicate that NOS may be translocated from the area associated with cytoplasmic organelles to cytosol following EC exposure to bradykinin.

Adult↗
PubMed1995

Ischemic time-dependent microvascular changes and reperfusion injury in the rat small intestine.

The significance of the reperfusion period in the pathophysiology of complete occlusion of the intestinal circulation is controversial. Our study was designed to evaluate the exact magnitude of reperfusion-induced intestinal mucosal damage in a standardized rat model of complete segmental arterial ischemia as a function of the occlusion time. Intestinal ischemia was maintained for 15, 30, or 60 min, or ischemia was followed by a 30-min reperfusion period. Intraarterial India ink perfusion was applied to visualize the mucosal vascularity changes induced by ischemia or ischemia-reperfusion. The height of the distributing arterioles of the villi and the average mucosal thickness were recorded by an image analysis system, and the degree of mucosal damage was established on a semiquantitative 0 to 5 grade scale. Ischemia induced erythrocyte obstructions at the villus tip, a progressive decrease in carbon-filled arteriole height, and a concomitant 7, 23, or 35% reduction of the mucosal thickness. The percentage decrease in perfused arteriole height/percentage mucosal thickness reduction ratio was 2.2, 1.5, or 1 during the 15-, 30-, or 60-min ischemia, respectively. Extravasation of the carbon tracer was observed in the 60-min ischemia group. During reperfusion, the mucosal layer was reduced by 27, 38, or 57%, respectively, compared with the baseline values. The arteriole height reduction/mucosal thickness reduction ratio was 1:1 in all ischemic-reperfused groups. The degree of mucosal damage was significantly increased during reperfusion after the 15-min ischemia. Microvessel obstruction is initiated at the villus tip following the onset of arterial occlusion, with subsequent destruction of the surrounding tissues during reperfusion. The reperfusion component of the net mucosal damage may be very significant in early forms of complete occlusion of the mesenteric circulation.

Animals↗
PubMed1995

Complete DNA sequence, specific Tn5 insertion map, and gene assignment of the carotenoid biosynthesis pathway of Rhodobacter sphaeroides.

The carotenoid biosynthesis genes form a cluster within the genome of Rhodobacter sphaeroides, lying in the middle of a larger cluster and 45 kb in length, which contains genes for bacteriochlorophyll biosynthesis and for the reaction center and light-harvesting apoproteins. The positions and approximate limits of the carotenoid genes were determined previously by localized transposon Tn5 mutagenesis and by comparison with the closely related Rhodobacter capsulatus carotenoid gene cluster. In this report, analysis of the DNA and deduced amino acid sequences of the carotenoid genes in R. sphaeroides are presented. Twenty-five Tn5 insertion mutants were used to produce a base-specific Tn5 insertion map of this region, and carotenoid gene assignment was supported by spectroscopic, ultrastructural, and high-pressure liquid chromatography analyses of these mutants. A region in the 3' end of crtD which affects bacteriochlorophyll biosynthesis was discovered, and CrtA was found to possess a proline-rich C-terminal region containing a repeated (Ala-Pro)n motif. CrtF also showed a high degree of sequence conservation with eukaryotic O-methyltransferases. This study provides gene sequences and assignments based upon a comprehensive structural, spectroscopic, and biochemical analysis of a range of carotenoid biosynthetic mutants; in each mutation, the point of Tn5 insertion is determined accurate to 1 bp on the gene cluster.

Amino Acid Sequence↗
PubMed1995

Response of mucosal mast cells to intestinal ischemia-reperfusion injury in the rat.

The goals of this study were to investigate the in vivo effects of intestinal ischemia-reperfusion on mucosal mast cells, and to evaluate the morphological changes induced by standardized arterial occlusion in anesthetized rats. Complete segmental ileal ischemia was maintained for 15, 30, or 60 min, and was followed by a 30 min reperfusion period. Intestinal biopsies taken at the end of ischemia and in the 30th min of reperfusion were evaluated by image analysis, and the rate of release of type II rat mast cell protease, a marker of mast cell exocytosis, was determined from the venous effluent of the segment. Electron microscopy revealed cytoplasmic vacuolization of the mast cells of the villi after the 15 min ischemia. Ischemia induced a continuous diminution of the mucosal thickness and a significant fall in the number of mast cells in the villi; with immunoperoxidase staining with a monoclonal antibody that recognizes the AD1 mast cell surface antigen, the decrease was 57, 49, and 66% in the 15, 30, and 60 min ischemia groups, respectively. In these groups, the mucosal type II mast cell protease concentration increased to 2.4-, 2.5-, and 3.6-fold, respectively, and a significant increase in plasma protease levels was observed on reperfusion. These results lead us to conclude that mucosal mast cells are very sensitive to intestinal ischemia, with the majority of mast cells in the ileal villi already involved in the response to ischemia after a short period of arterial occlusion.

Animals↗
PubMed1994

Properties of synthesis of chlorophyll a from chlorophyll b in cucumber etioplasts.

Chlorophyll a accumulated when chlorophyllide b was incubated with isolated cucumber etioplasts in the dark. When [14C]chlorophyllide b was used as the substrate for chlorophyll synthesis, [14C]chlorophyll a was formed, showing that chlorophyll a was synthesized from the exogenously added chlorophyllide b, not by light-independent reduction of endogenous protochlorophyllide. The reaction studied showed an optimal pH of 7.5 and required both the soluble and membrane fractions of etioplasts together with ATP. Time course experiments showed that chlorophyll a began to accumulate later than chlorophyll b. Chlorophyllide b esterified with geranylgeraniol accumulated first, but there was little accumulation of chlorophyll a with an unhydrogenated prenyl side chain. No chlorophyllide a was detected during incubation. These observations indicate that esterified chlorophyll b was converted to chlorophyll a. This conversion would play an important role in the use of chlorophyll b for the synthesis of chlorophyll a in the reconstruction of photosystems.

Chlorophyll↗
PubMed1994

Reconstitution of import-competent outer membrane vesicles from mammalian mitochondria.

Protein insertion into mitochondrial outer membrane (OM) vesicles isolated from Neurospora crassa has recently been reported. The N. crassa OM vesicles retained the features of the intact mitochondria concerning the dependency of insertion on the receptor protein [A. Mayer et al. (1993) J. Cell Biol. 121, 1233-1243]. In this study, OM vesicles were purified from bovine adrenal cortex mitochondria, and unilamellar proteoliposomes were reconstituted from OM vesicles using heptyl beta-thioglucoside. Both OM vesicles and the reconstituted outer membrane vesicles (ROM) were able to import porin, but unable to import the precursor of adrenodoxin, which translocates across both the outer and inner membranes of intact mitochondria. Porin insertion into both OM vesicles and ROM was inhibited in the presence of purified recombinant adrenodoxin precursor and also by ATP depletion, and was dependent on the trypsin-sensitive membrane surface factor, suggesting that the purified OM vesicles as well as ROM retained the properties of the intact OM concerning porin insertion. The protein import machinery of OM seems to be functional for the outer membrane protein without the participation of the inner membrane. The successful reconstitution of the protein import activity from solubilized OM will pave the way for further biochemical characterization of the protein import machinery of OM.

Adenosine Triphosphate↗
PubMed1994

Introduction of new carotenoids into the bacterial photosynthetic apparatus by combining the carotenoid biosynthetic pathways of Erwinia herbicola and Rhodobacter sphaeroides.

Carotenoids have two major functions in bacterial photosynthesis, photoprotection and accessory light harvesting. The genes encoding many carotenoid biosynthetic pathways have now been mapped and cloned in several different species, and the availability of cloned genes which encode the biosynthesis of carotenoids not found in the photosynthetic genus Rhodobacter opens up the possibility of introducing a wider range of foreign carotenoids into the bacterial photosynthetic apparatus than would normally be available by producing mutants of the native biosynthetic pathway. For example, the crt genes from Erwinia herbicola, a gram-negative nonphotosynthetic bacterium which produces carotenoids in the sequence of phytoene, lycopene, beta-carotene, beta-cryptoxanthin, zeaxanthin, and zeaxanthin glucosides, are clustered within a 12.8-kb region and have been mapped and partially sequenced. In this paper, part of the E. herbicola crt cluster has been excised and expressed in various crt strains of Rhodobacter sphaeroides. This has produced light-harvesting complexes with a novel carotenoid composition, in which the foreign carotenoids such as beta-carotene function successfully in light harvesting. The outcome of the combination of the crt genes in R. sphaeroides with those from E. herbicola has, in some cases, resulted in an interesting rerouting of the expected biosynthetic sequence, which has also provided insights into how the various enzymes of the carotenoid biosynthetic pathway might interact. Clearly this approach has considerable potential for studies on the control and organization of carotenoid biosynthesis, as well as providing novel pigment-protein complexes for functional studies.

Carotenoids↗
PubMed1993

Human neonatal and adult vascular smooth muscle cells in culture.

Neonatal vascular smooth muscle cells (SMC) in culture have been demonstrated to be quite different from adult SMC and to be similar to intimal SMC in animal models. To characterize human neonatal vascular SMC in culture, cultures of arterial SMC were prepared by an explant method from the subclavian arteries of autopsied patients (10 adults and 6 neonates). The morphology and growth characteristics of these cells were compared. All cells were positively immunostained with HHF 35, a monoclonal antibody specific for muscle actin. Electron microscopically, both adult and neonatal SMC were of synthetic phenotype. SMC from neonates had a short population doubling time (PDT, 28.6 +/- 7.5 hr) and high saturation density (SD, 37.5 +/- 11.9 x 10(4) cells/cm2). They did not show hill and valley growth patterns. Among the SMC cultured from adult media, two subtypes were distinguished, based on their growth characteristics. Classical adult SMC (7 of 10 cases) grew in hill and valley patterns with long PDT and low SD values (61.7 +/- 28.8 hr, 6.5 +/- 1.9 x 10(4) cells/cm2, respectively). The second subtype (3 of 10 cases), neonatal-type adult SMC, had PDT and SD values (22.9 +/- 4.0 hr, 31.3 +/- 14.7 x 10(4) cells/cm2, respectively) similar to those of neonatal SMC. Intimal SMC became senescent in early phases of subculture. To test for the possible participation of autocrine growth factors in the heterogeneity of the growth patterns, Northern blot analysis was conducted for PDGF-A, TGF-beta, c-myc, and c-fos mRNA in three types of SMC. There was no significant difference in these mRNA levels between the SMC. We demonstrated that human neonatal vascular SMC in culture are quite different in their growth characteristics from classical adult SMC in culture and that neonatal-type SMC can be isolated from adult media.

Adult↗