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41.
Noriyuki Suka Yoshinobu Shinohara Yasushi Saitoh Hisato Saitoh Kohei Ohtomo Masahiko Harata Edward Shpigelman Shigeki Mizuno 《Genetica》1993,88(2-3):93-105
About 65% of DNA in the chicken W chromosome has been shown to consist ofXhoI andEcoRI family repetitive sequences. These sequences showed remarkable delay in the electrophoretic mobility at low temperature
on a polyacrylamide gel. Three dimensional structures of the 0.7-kbXhoI and the 1.2-kbEcoRI family repeating units were estimated to be irregular solenoids using a computer program based on wedge angles of all the
16 dinucleotide steps. Fluorescencein situ hybridization demonstrated that these two family sequences were localized in a major heterochromatic body in an interphase
nucleus. Incorporation of bromodeoxyuridine into the W chromosome in the synchronous culture of MSB-1 cells occurred about
1 h later than the peak of S phase. The chromatin structure formed alongXhoI andEcoRI family sequences was suggested to be different from the total chromatin or chromatin containing the β-actin gene sequence
in that the linker DNA lengths of the former were significantly longer. Fractionation of theHaeIII-digested MSB-1 nuclei yielded a chromatin fraction in whichXhoI family sequences were partially enriched. Several DNA-binding proteins showing higher affinity for theXhoI family sequence were present in this fraction. 相似文献
42.
Yasushi Sato Munetaka Sugiyama Ryszard J. Górecki Hiroo Fukuda Atsushi Komamine 《Planta》1993,189(4):584-589
In a culture system in which single cells isolated from the mesophyll of Zinnia elegans L. differentiate to tracheary elements (TEs), two inhibitors of phenylalanine ammonia-lyase (EC 4.3.1.5), L-α-aminooxy-β-phenylpropionic acid (AOPP) at 10 μM inhibited lignification without reducing the number of TEs formed. These inhibitors caused intracellular changes in peroxidase (EC 1.11.1.7) activities. The inhibitors increased the activity of peroxidases bound to the cell walls and especially the activity of peroxidase bound ionically to the cell walls. In contrast, the activity of extracellular peroxidase decreased. There were five isoenzymes, P1-P5, in the ionically bound peroxidase of cultured Zinnia cells. Among the isoenzymes, P4 and P5 appeared to be specific for TE differentation. Treatment with AOPP and AIP resulted in increases in the activities of P2, P4 and P5 isoenzymes, with the most prominent increase in P5 activity. The addition of lignin precursors, including coniferyl alcohol, to the AOPP-treated cells restored lignification, and suppressed the alteration of peroxidase isoenzyme patterns caused by AOPP. The relationship between the wall-bound peroxidases and lignification during TE differentiation is discussed in the light of these results. 相似文献
43.
Kazuyoshi Masuda Shunji Nagata Koichiro Hirano Yasushi Takagishi Hidematsu Hirai 《Microbiology and immunology》1993,37(2):165-167
This paper describes an attempt to effectively induce antibody-dependent cell-mediated cytotoxicity (ADCC) in nude mice. A monoclonal antibody against α-fetoprotein, 80G, coadministered with spleen cells from other nude mice bearing HuH-7N (xenograft of human hepatoma cell line, HuH-7) significantly suppressed the growth of HuH-7N as compared to treatment with 80G alone. 80G with spleen cells from normal nude mice also had some suppressive effect. In contrast, no effect was observed with each spleen cells alone as well as 80G alone. These results suggest that further supply of effector cells could enhance ADCC activity in nude mice. 相似文献
44.
Hideaki Matsuoka Yasushi Kazuno Takuji Horie Tomoo Homma Yasuyuki Nemoto 《Cytotechnology》1993,11(1):59-65
Possible roles of coexisting cells in inducing neurite growth from a nerve cell were studied. Nerve growth factor (NGF)-inducing neurite growth from PC12h-R (a cell line derived from cultured nerve cells) was investigated at various cell densities. At the cell density 102104 cells/ml neurites appeared even without NGF. In contrast, no neurite appeared without NGF in single cell culture. The neurite growth observed in plural cell culture without NGF was only partially inhibited by antibody to NGF receptor (Ab-NGFR). However, the effect of the used medium alone was mostly inhibited by Ab-NGFR. These results suggest that the neurite inducing potency of coexisting cells is via different sites than the NGF receptor.Abbreviations Ab-IgG-FITC
anti-mouse-IgG labeled with fluorescein isothiocyanate
- Ab-NF
monoclonal antibody to neurofilament 160 kD
- Ab-NGFR
monoclonal antibody to NGF receptor
- BDNF
brain-derived neurotrophic factor
- D-medium
medium for differentiation culture
- DMEM
Dulbecco's modified Eagle's medium
- M-medium
medium for multiplication culture
- NGF
nerve growth factor
- NGFR
NGF receptor
- NT-3
neurotrophin-3
- PC12
pheochromocytoma cell line
- PC12h-R
subclone of PC12
- Sup-D
supernatant of D-medium 相似文献
45.
Kazuyoshi Masuda Shunji Nagata Koichiro Hirano Yasushi Takagishi Hidematsu Hirai 《生物化学与生物物理学报:疾病的分子基础》1993,1182(2):128-132
This study was carried out to clarify the reason for elevation of serum α-fetoprotein (AFP) level of nude mice bearing hepatoma cells after treatment with monoclonal antibodies (MoAbs) to AFP. MoAbs to AFP showed no effect on the cumulative amounts of AFP secreted from human hepatoma cell line, HuH-7, in vitro. However, the treatment of nude mice bearing HuH-7N cells (HuH-7 xenograft) with MoAbs to AFP led to elevation of the serum AFP level in spite of the fact that the growth curve of HuH-7N cells was similar to that for PBS treatment. This apparent elevation of the serum AFP level is thought to be due to the slow elimination of AFP-MoAb immune complexes with little lattice structure from circulation, but not the enhancement of AFP secretion of HuH-7N cells. Thus, when using a MoAb alone or MoAb-drug conjugate, the serum AFP level should only be cautiously used as a tumor marker for evaluating the targeting immunotherapy. 相似文献
46.
Koichi Matsuzaki Mikio Kan Wallace L. McKeehan 《In vitro cellular & developmental biology. Animal》1996,32(6):345-360
Summary Two transmembrane serine-threonine kinases (type I and II receptors), a membrane-anchored proteoglycan (type III), and a homodimeric
ligand participate in the transforming growth factor beta type on (TGFβ1) signal transduction complex. The expression of recombinant
receptors in insect cells co-infected with up to three recombinant baculoviruses was employed to study interactions among
the ectodomains of the three types of receptors and the TGFβ1 ligand in absence of uncontrollable extrinsic factors in mammalian
cells. Multi-subunit complexes were assembled in intact cells and purified on glutathione-conjugated beads for analysis by
tagging one of the subunits with glutathione S-transferase (GST). Intrinsic ligand-independent interactions were observed
among receptor subunits as follows: type III–III type I–I, type III-I, and type II-I. The homeotypic complex of type II–II
receptors and the heterotypic type III-II interaction was ligand dependent. The type I, but not the type III, subunit displaced
about 50% of the type II component in either ligand-dependent homomeric type II-type II complexes or heteromeric type III-type
II complexes to form type II-I or type III-II-I oligomers, respectively. The type II subunit displaced type I subunits in
oligomers of the type I subunit. Specificity of type I receptors may result from differential affinity for the type II receptor
rather than specificity for ligand. A monomeric subunit of the TGFβ1 ligand bound concurrently to type III and type II or
type III and type I receptors, but failed to concurrently bind to the type II and type I subunits. The binding of TGFβ1 to
the type I kinase subunit appears to require an intact disulfide-linked ligand dimer in the absence of a type III subunit.
The combined results suggest a pentameric TGFβ signal transduction complex in which one unit each of the type III, type II,
and type I components is assembled around the two subunits of the dimeric TGFβ1 ligand. An immobilized GST-tagged subunit
of the receptor complex was utilized to assemble multi-subunit complexesin vitro and to study the phosphorylation events among subunits in the absence of extrinsic cell-derived kinases. The results revealed
that (a) a low level of ligand-independent autophosphorylation occurs in the type I kinase; (b) a high level of autophosphorylation
occurs in the type II kinase; (c) both the type III and type I subunits aretrans-phosphorylated by the type II subunit; and (d) the presence of both type I and II kinases complexed with the type III subunit
and dimeric TGFβ1 ligand in a pentameric complex causes maximum phosphorylation of all three receptor subunits. 相似文献
47.
Shigeko Ishimatsu Toshihiro Kawamoto Koji Matsuno Yasushi Kodama 《Biological trace element research》1995,49(1):43-52
In this study, eight kinds of nickel (Ni) compounds were orally administered to Wistar male rats and the distribution of each
compound was investigated 24 h after the administration. The Ni compounds used in this experiment were nickel metal [Ni−M],
nickel oxide (green) [NiO(G)], nickel oxide (black) [NiO(B)], nickel subsulfide [Ni3S2], nickel sulfide [NiS], nickel sulfate [NiSO4], nickel chloride [NiCl2], and nickel nitrate [Ni(NO3)2]. The solubilities of the nickel compounds in saline solution were in the following order; [Ni(NO3)2>NiCl2>NiSO4]≫[NiS>Ni3S2]>[NiO(B)>Ni−M>NiO(G)]. The Ni level in the visceral organs was higher in the rats given soluble Ni compounds; Ni(NO3)2, NiCl2, NiSO4, than that in the rats receiving other compounds. In the rats to which soluble Ni compounds were administered, 80–90% of
the recovered Ni amounts in the examined organs was detected in the kidneys. On the other hand, the Ni concentration in organs
administered scarcely soluble Ni compounds; NiO(B), NiO(G), and Ni−M were very low. The estimated absorbed fraction of each
Ni compounds was increased with the increase of the solubility. These results suggest that the kinetic behavior of Ni compounds
administered orally is closely related with the solubility of Ni compounds, and that the solubility of Ni compounds is one
of the important factors for determining the health effect of Ni compounds. 相似文献
48.
Changes in chorion proteins induced by the exudate released from the egg cortex at the time of fertilization in the teleost, Oryzias latipes 总被引:2,自引:2,他引:0
Takashi Iwamatsu Yasushi Shibata Toshihiro Kanie 《Development, growth & differentiation》1995,37(6):747-759
The chorion of unfertilized medaka Oryzias latipes eggs consists of two major proteins (77–73 and 49 kDa) and a minor 150 kDa protein. Upon fertilization, these major chorion proteins are polymerized to insoluble high molecular weight proteins via the temporary formation of several new proteins (132, 114, 62 and 61 kDa). Increasing chorion toughness is closely related to the formation of high molecular weight proteins and the increasing insolubility of the chorion proteins. The changes in chorion proteins and hardening could be induced in vitro in isolated chorions by an egg exudate, which includes cortical alveolar contents. The effects of temperature and pH on the egg exudate-induced changes in chorion proteins were examined in the present study. The major proteins could be digested by proteolytic enzymes. The 49 kDa protein was PAS-positive. Analysis with polyclonal antibodies against the major proteins demonstrated that the temporarily formed 62 and 61 kDa proteins were derived from the 77–73 kDa protein and that higher molecular weight proteins, newly formed in the process of chorion hardening, contained the same epitopes as did the 77–73 and 49 kDa proteins. The results suggest that the changes in chorion proteins of the medaka egg at the time of fertilization can be induced by an enzyme(s) released from the egg cortex into the perivitelline space. 相似文献
49.
In vitro 5-day cultures of naive spleen cells with viable Listeria monocytogenes (VLM), but not heat-killed L. monocytogenes, induced CD4+ T cells that produced IFN-γ upon secondary antigen stimulation. The VLM-induced Listeria-specific T cells produced IFN-γ but lacked expression of IL-2 and IL-4. To study the role of IFN-γ in the induction of the IFN-γ-producing T cells, we added anti-IFN-γ mAb to the primary culture and analyzed IFN-γ production upon secondary antigen stimulation. Addition of anti-IFN-γ mAb to the culture suppressed generation of IFN-γ-producing CD4+ T cells, suggesting that IFN-γ is important in the induction of IFN-γ-producing CD4+ T cells. Furthermore, our results showed that depletion of NK cells from spleen cells by anti-asialo GM1 antibody plus complement before culture enhanced induction of IFN-γ-producing CD4+ T cells. Although NK cells are known to produce IFN-γ, the results indicate that NK cell-derived IFN-γ may not be important in induction of the Listeria-specific IFN-γ-producing CD4+ T cells in the culture system. In addition, we demonstrated that IFN-γ expression was high in CD4+ T cells from cultures of spleen cells with VLM at the primary culture level. These results suggest that IFN-γ derived from T cells may enhance production of IFN-γ by CD4+ T cells, while NK cells rather suppress the induction of IFN-γ-producing CD4+ T cells. 相似文献
50.