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171.
We examined the effect of methionine deficiency on iodothyronine 5’-deiodinase activity in selenium-deficient rats or selenium-sufficient
rats fed sodium selenate or selenomethionine. Forty-two weanling male Wistar rats were divided into six groups and pair fed
the respective purifiedl-amino acid-based diets for 4 wk.l-methionine concentrations in the diet were 8.0 g/kg for sufficient rats, and 2.0 g/kg for deficient rats. Selenium concentrations
in the diet were 0.5 mg/kg (as sodium selenate or selenomethionine) for selenium-sufficient rats and less than 0.005 mg/kg
for selenium-deficient rats. Type I 5’-deiodinase activities were significantly lower in liver and higher in kidney of methionine-deficient
rats than in those of methionine-sufficient rats fed either the selenium-sufficient or the selenium-deficient diets. The type
I 5’-deiodinase activity in brain was significantly lower in the methionine-deficient rats than in the methionine-sufficient
rats fed the selenium-deficient diet. Type II 5’-deiodinase activity in brain was significantly higher in the methionine-deficient
rats than in the methionine-sufficient rats fed selenium-sufficient diet as sodium selenate. Both thyroxine and 3,3’,5-triiodothyronine
concentrations in plasma were significantly higher in the methionine-deficient rats than in the methionine-sufficient rats.
It is suggested that the methionine deficiency affects the 5’-deiodinase activity and thyroid hormones level in the rats. 相似文献
172.
The N-terminal region of hepatitis C virus nonstructural protein 3 (NS3) is essential for stable complex formation with NS4A. 总被引:11,自引:9,他引:2
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Hepatitis C virus proteins are produced by proteolytic processing of the viral precursor polyprotein that is encoded in the largest open reading frame of the viral genome. Processing of the nonstructural viral polyprotein requires the viral serine-type proteinase present in nonstructural protein 3 (NS3). The cleavage of the junction between NS4B and NS5A is mediated by NS3 only when NS4A is present. NS4A is thought to be a cofactor that enhances the cleavage efficiency of NS3 in hepatitis C virus protein-producing cells. Stable NS3-NS4A complex formation required the N-terminal 22 amino acid residues of NS3. This interaction contributed to stabilization of the NS3 product as well as increased the efficiency of cleavage at the NS4B/5A site. The N-terminal 22 amino acid residues fused to Escherichia coli dihydrofolate reductase also formed a stable complex with NS4A. NS3 derivatives which lacked the N-terminal 22 amino acid residues showed drastically reduced cleavage activity at the NS4B/5A site even in the presence of NS4A. These data suggested that the interaction with NS4A through the 22 amino acid residues of NS3 is primarily important for the NS4A-dependent processing of the NS4B/5A site by NS3. 相似文献
173.
Cloning and characterization of a gene involved in aerial mycelium formation in Streptomyces griseus.
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A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) is essentially required for aerial mycelium formation and streptomycin production in Streptomyces griseus. A DNA fragment which induced aerial mycelium formation and sporulation in an A-factor-deficient mutant strain, S. griseus HH1, was cloned from this strain on a high-copy-number plasmid. Subcloning and nucleotide sequencing revealed that one open reading frame with 218 amino acids, named AmfC, served as a multicopy suppressor of the aerial mycelium-defective phenotype of the A-factor-deficient strain. The amfC gene did not restore A-factor or streptomycin production, indicating that amfC is involved in aerial mycelium formation independently of secondary metabolic function. Disruption of the chromosomal amfC gene in the wild-type S. griseus strain caused a severe reduction in the abundance of spores but no effect on the shape or size of the spores. The infrequent sporulation of the amfC disruptant was reversed by introduction of amfC on a plasmid. The amfC-defective phenotype was also restored by the orf1590 gene but not by the amfR-amfA-amfB gene cluster. Nucleotide sequences homologous to the amfC gene were distributed in all of 12 Streptomyces species tested, including Streptomyces coelicolor A3(2). The amfC homolog of S. coelicolor A3(2) was cloned and its nucleotide sequence was determined. The AmfC products of S. griseus and S. coelicolor A3(2) showed a 60% identity in their amino acid sequences. Introduction of the amfC gene of S. coelicolor A3(2) into strain HH1 induced aerial mycelium formation and sporulation, which suggests that both play the same functional role in morphogenesis in the strains. 相似文献
174.
Putative morphogen, DIF, of Dictyostelium discoideum induces apoptosis in rat pancreatic AR42J cells
Yuzuru Kubohara Chiharu Kimura Kazuhiko Tatemoto 《Development, growth & differentiation》1995,37(6):711-716
We have recently shown that differentiation-inducing factor-1 (DIF-1) of Dictyostelium discoideum is capable of raising intracellular calcium concentration ([Ca2+ ]i ) and suppressing cell proliferation of rat pancreatic AR42J cells in a dose-dependent manner, and that DIF-1 at a concentration of 40 μmol/L is toxic to the cells. In this study, we have further characterized the cytotoxic effect of DIF-1 on AR42J cells and have analyzed the effect of DIF-1 on [Ca2+ ]i . In the presence of 40 μmol/L DIF-1, cells began to bleb after approximately 6 h, and most had died within 48 h. Biochemical analysis revealed that DNA fragmentation was accompanied by cell death. Monitoring the changes in [Ca2+ ]i induced by DIF-1, it was found that cells were able to adapt to stimulation with DIF-1 so that they did not respond to subsequent stimulation by DIF-1. These results indicate that DIF-1 induced apoptosis in AR42J cells probably via a cell signaling system. 相似文献
175.
The level of aggressiveness and the weight of preputial gland and testis in male mice (Mus musculus) were influenced by housing condition, especially by the presence of cohabitant males. In this study, the relation between
aggressiveness and the preputial gland and testis weight was studied for various housing conditions. The mouse individually
housed in a cage that was linked to another cage containing another male separated by wire net was more aggressive than isolated
or paired mice. The preputial gland weight also showed the same tendency, suggesting that the odor from other males promotes
pituitary-gonadal activity in males, and that long-term cohabitance inhibits it. 相似文献
176.
Y. Kobayashi Yufuko Takahashi Satoshi Chikayama Motomi Ikeda Nobuhiko Uoshima Shinya Kimura Koji Tanaka Katuya Wada Masaru Ozawa Tatuo Sugano Naoyuki Maruo Motoharu Kondo 《Histochemistry and cell biology》1997,108(2):115-120
We devised a new microfluorometric method for determining the ploidy of megakaryocytes identified immunologically in bone
marrow smears. The smears were immunostained by incubation with mouse monoclonal anti-glycoproteins (GP) IIb antibodies, followed
by fluorescein isothiocyanate-conjugated goat anti-mouse IgG antibodies. They were then stained with 4′,6-diamidino-2-phenylindole
(DAPI). Megakaryocytes were identified by their GPIIb immunofluorescence using a microfluorometer and, after the filters were
changed, their DNA content was assayed by measuring the intensity of DAPI fluorescence. This intensity was shown to be proportional
to the DNA content when the aperture of the objective lens was reduced. We compared these results with those obtained when
megakaryocytes were identified morphologically, using DAPI staining after Wright-Giemsa destaining. In all 12 normal controls,
the ploidy peaks were shown to be 16N by both methods, and the mean ploidy detected by the immunological method was only reduced
0.961 times relative to the estimate from the morphological method. In contrast, in eight myelodysplastic syndrome (MDS) patients,
the ploidy peaks were either 8N or 4N and the mean was reduced by 0.906 times (P=0.018). Thus we could immunologically identify small megakaryocytes which we could not identify morphologically. Therefore,
this method is useful for measuring megakaryocytic ploidy, especially in the pathological megakaryocytes of MDS patients.
Accepted: 29 April 1997 相似文献
177.
Elucidation of the mechanism enhancing the avidity of lectin with oligosaccharides on the solid phase surface 总被引:3,自引:2,他引:1
The mechanism underlying molecular recognition of lectins waselucidated by a novel solid phase binding assay system basedon surface plasmon resonance. When the apparent affinities ofinteractions between chitooligosaccharides and wheat germ agglutininwere compared between lectin-immobilized and oligosaccharide-immobilizedassay systems, the affinity constants (Ka) calculated for theformer system were in good agreement with the previously reportedvalues measured in solution. On the other hand, in the lattersystem, the calculated Ka could be more than 10,000 times higherthan the values in solution at lower lee tin concentrations.To elucidate the reason for this, we systematically investigatedthe effects of the oligosaccha-ride immobilized density andthe lectin valence on the apparent affinity in the oligosaccharide-immobilizedassay system. Both the apparent association (kass) and dissociationrate constants (kdiss) showed a tendency to decrease as theoligosaccharide density increased. This effect was most remarkablefor the interaction possessing an extremely fast intrinsic Kass.Oligomerization of lectin enhanced the avidity due to a significantreduction in kdiss. These phenomena could be explained by consideringthe nonhomogeneous conditions under which binding occurred.The reaction in a nonhomogeneous state is limited by the masstransport effect, and the effect of rebinding becomes so largethat it cannot be disregarded. These findings are the firstto demonstrate the importance of the mass transport effect inmodulating the affinity of lectin for oligosaccharides on asolid phase surface. avidity clustering effect lectin mass transport surface plasmon resonance 相似文献
178.
Souichi Satoh †Tatsuo Kimura †Masahiro Toda †Mutuko Maekawa †Satoshi Ono †Hirokazu Narita Hiroyuki Miyazaki Toshihiko Murayama Yasuyuki Nomura 《Journal of neurochemistry》1997,69(5):2197-2205
Abstract: Nitrogen oxides, such as nitric oxide, have been shown to regulate neuronal functions, including neurotransmitter release. We investigated the effect of S-nitroso-l -cysteine (SNC) on noradrenaline (NA) release in the rat hippocampus in vivo and in vitro. SNC stimulated [3H]NA release from prelabeled hippocampal slices in a dose-dependent manner. SNC stimulated endogenous NA release within 30 min to almost five times the basal level in vivo (microdialysis in freely moving rats). In a Na+-containing Tyrode's buffer, SNC-stimulated [3H]NA release was inhibited 30% by the coaddition of l -leucine. In the Na+-free, choline-containing buffer, SNC-stimulated [3H]NA release, which was similar to that in the Na+-containing buffer, was inhibited markedly by l -leucine, l -alanine, l -methionine, l -phenylalanine, and l -tyrosine. The effects of the other amino acids examined were smaller or very limited. The effect of l -leucine was stronger than that of d -leucine. A specific inhibitor of the L-type amino acid transporter, 2-aminobicyclo[2.2.1]-heptane-2-carboxylate (BCH), inhibited the effects of SNC on [3H]NA release in the Na+-free buffer. Uptake of l -[3H]leucine into the slices in the Na+-free buffer was inhibited by SNC, BCH, and l -phenylalanine, but not by l -lysine. The effect of SNC on cyclic GMP accumulation was not inhibited by l -leucine, although SNC stimulated cyclic GMP accumulation at concentrations up to 25 µM, much less than the concentration that stimulates NA release. These findings suggest that SNC is incorporated into rat hippocampus via the L-type-like amino acid transporter, at least in Na+-free conditions, and that SNC stimulates NA release in vivo and in vitro in a cyclic GMP-independent manner. 相似文献
179.
Mutational analysis of human immunodeficiency virus type 1 (HIV-1) accessory genes: requirement of a site in the nef gene for HIV-1 replication in activated CD4+ T cells in vitro and in vivo.
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Y Kawano Y Tanaka N Misawa R Tanaka J I Kira T Kimura M Fukushi K Sano T Goto M Nakai T Kobayashi N Yamamoto Y Koyanagi 《Journal of virology》1997,71(11):8456-8466
Human immunodeficiency virus type 1 (HIV-1) accessory genes including nef, vif, and vpr are important factors that determine the replication and pathogenesis of HIV-1. The state of activation is also important for the replication of HIV-1. We evaluated the properties of nef-, vif-, and vpr-minus macrophage-tropic HIV-1(JR) CSF in primary CD4+ Th1- or Th2-like cell cultures which had been activated through CD3 molecules in the presence of interleukin-2 (IL-2) and IL-12 (Th1-like culture) or IL-4 (Th2-like culture), respectively. In activated Th1- or Th2-like cultures, replication of nef-minus HIV-1(JR-CSF) was markedly lower than that of wild-type HIV-1. Subsequent analysis by site-directed mutagenesis showed that (i) the presence of an acidic amino acid-rich domain (amino acid residues 72 to 75) in the Nef protein was critical for the enhancement of viral DNA synthesis, resulting in increased virus growth rate, and (ii) prolines that form part of Src homology 3 binding domain were not essential for viral replication. We also confirmed the importance of sites by using an HIV-1-infected animal model, the hu-PBL-SCID mouse system, representing HIV-1 replication and pathogenesis in activated CD4+ T cells in vivo. These results indicate that Nef accelerates viral replication in activated CD4+ T cells. 相似文献
180.
Clonal expansion of superantigen-reactive T cells is resistant to FK506 in mice with AIDS.
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Subcutaneous injection of FK506 (10 mg/kg of body weight) completely blocked the clonal expansion of staphylococcal enterotoxin A (SEA)-reactive T cells in healthy (control) mice after SEA injection but did not disturb it in mice with murine AIDS (MAIDS) caused by infection with LP-BM5 murine leukemia virus. MAIDS mice are characterized by utilization of a FK506-insensitive pathway for clonal expansion of superantigen-reactive T cells. 相似文献