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11.
12.
Serum-free culture of rat keratinocytes 总被引:2,自引:0,他引:2
Hirosuke Oku Chikara Kumamoto Tomoyuki Miyagi Takanori Hiyane Junichi Nagata Isao Chinen 《In vitro cellular & developmental biology. Animal》1994,30(8):496-503
Summary Procedures for the serum-free culture of rat keratinocytes have been established. Basal cells prepared from epidermis of newborn
rat were stored in liquid nitrogen and used for primary culture. Among the available media, MCDB 153, developed originally
for human keratinocyte (HK) culture, was the best for the development of serum-free formulation. To grow rat keratinocytes,
bovine serum albumin was arbitrarily substituted for the macromolecule supplements needed for HK culture, i.e. fetal bovine
serum protein or bovine pituitary extract. Qualitative and quantitative adjustment of supplements was thereafter made to support
rapid cell growth. Satisfactory cell growth was achieved in the optimized medium of MCDB 153 supplemented with growth factors
and amino acids: insulin (10 μg/ml), hydrocortisone (0.1 μg/ml), epidermal growth factor (25 ng/ml), calcium chloride (0.2
mM), histidine (0.23 mM), isoleucine (0.05 mM), tryptophane (0.015 mM), threonine (1.25 mM), tyrosine (0.031 mM), alanine (4.08 mM), and albumin (2 mg/ml). This optimized culture system was superior to the original HK culture condition for rapid growth
of rat keratinocytes. Under our condition, cells grew as a monolayer, becoming confluent, but without stratification, and
were passaged 2 to 3 times without any changes in morphology. The serum-free formulation allows us to control more accurately
the concentrations of biomolecules in the medium including lipids and hormones, and therefore will be suitable for the study
focusing on lipid metabolism or hormonal regulation of rat keratinocytes. 相似文献
13.
14.
Minoru Niimoto Takao Hattori Ichiji Ito Ryuichiro Tamada Kiyoshi Inokuchi Kunzo Orita Hisashi Furue Nobuya Ogawa Tomohiro Toda Motonosuke Furusawa Shigemasa Koga Isamu Hashimoto Tatsuhei Kondo Shigeru Fujimoto Yuzuru Sugiyama Osahiko Abe Masaaki Oya 《Cancer immunology, immunotherapy : CII》1984,18(1):13-18
Summary The usefulness of LMS in postoperative immunochemotherapy of gastric cancer was investigated. In compliance with the protocol, MMC was given at a dose of 20 mg on the day of gastrectomy, and an additional 10 mg on the next day IV. The patients receiving 600 mg Tegafur daily were then divided into two groups according to whether LMS was also given or not. LMS was administered for 3 days before the operation in a daily dose of 150 mg and for 1 year or more after operation according to a schedule of 3 days' administration followed by an 11-day interval. The 2-year follow-up demonstrated that in stage III patients, the LMS (+) regimen was superior to the LMS (–) regimen, since the former prolonged the relapse-free interval significantly. The survival rate for stage III disease was also significantly higher in the LMS (+) than in the LMS (–) group. There was no significant difference in the incidence of subjective or objective side-effects between two groups. The incidence of agranulocytosis was comparable in the two groups.Gastrointestinal Cancer Research Group, Japan Levamisole Research AssociationChairmen of the Gastrointestinal Cancer Research Group, Japan LMS Research AssociationController of the Gastrointestinal Cancer Research Group, Japan LMS Research AssociationMembers of the Data Collection and Analysis SubcommitteeThis study was carried out by the Gastrointestinal Cancer Research Group, Japan LMS Research Association (directed by Prof. Kiyoshi Inokuchi, Dept. of Surgery, Kyushu University and Prof. Eiro Tsubura, Dept. of Internal Medicine, Tokushima University). The results were presented in part at the 19th General Meeting of the Japanese Society for Gastroenterological Surgery in February, 1982 相似文献
15.
A new synthesis of desmosterol was described using hyodeoxycholic acid (3,6-dihydroxy-5β-cholanic acid) as a starting material. Epidesmosterol (3-hydroxycholesta-5,24-diene) was also synthesized for the first time from the same starting material. 相似文献
16.
M Iwakura K Furusawa T Kokubu S Ohashi Y Tanaka Y Shimura K Tsuda 《Journal of biochemistry》1992,111(1):37-45
Dihydrofolate reductase (DHFR) has been demonstrated to be a versatile "affinity handle" for expression of recombinant proteins. The DHFR "handle" has advantages not only in terms of efficiency of expressing the fusion protein as a soluble form but also in stabilizing unstable polypeptides and facilitating purification of the expressed protein by means of methotrexate-bound affinity chromatography and by making use of the enzyme activity. Fifteen genes encoding different lengths of polypeptides of 5 to 44 amino acids were chemically synthesized and introduced into expression vectors, pTP70-1 or its derivatives. All the polypeptide genes were efficiently expressed in Escherichia coli cells as fusion proteins which show DHFR activity. The respective fusion proteins were highly purified from cell-free extracts by monitoring the DHFR activity at each purification step. The use of methotrexate-bound affinity chromatography was very effective. In order to cut out the polypeptides, the purified fusion proteins were treated with either BrCN or site-specific protease according to the spacer sequence. The objective polypeptide was purified by means of a reversed-phase high-pressure liquid chromatography (HPLC) system. Specific cleavage of the purified fusion protein actually yielded very few peptide fragments, so the assignment and isolation of the objective polypeptide were carried out without difficulty. 相似文献
17.
Masanao Watanabe Keiji Itoh Kuniya Abe Toshifumi Akizawa Kohji Ikenishi Mitsuru Furusawa 《Development, growth & differentiation》1992,34(2):223-231
In order to investigate whether a vasa -like protein is present in germ line cells of Xenopus , antibodies were produced which react specifically with synthetic oligopeptides of sequences from near the N- or C-termini or with one including the DEAD box of the Drosophila vasa protein.
Only the antibody against the oligopeptide including the DEAD box reacted strongly with germ plasm (GP) or with cytoplasm of germ line cells of Xenopus embryos by immunofluorescence microscopy. By immunoelectron microscopy, the antibody was demonstrated to react with the GP-specific structure, germinal granules, in cleaving embryos, and with their derivatives in the germ line cells of embryos at stages extending from gastrula to feeding tadpole. It also reacted with mitochondria not only in the GP and the germ line cells but also in somatic cells, and with myofibrils in muscle cells. By Western blotting, the antibody was shown to react with several bands of Mr 42–69 ± 103 in protein samples from Xenopus embryos. In samples from Drosophila ovaries, it reacted with a Mr 71 ± 103 band which was probably the vasa protein. This indicates the possibility that Xenopus embryos contain several DEAD family proteins. One of these is present on germinal granules, resembling the vasa protein on polar granules of Drosophila . 相似文献
Only the antibody against the oligopeptide including the DEAD box reacted strongly with germ plasm (GP) or with cytoplasm of germ line cells of Xenopus embryos by immunofluorescence microscopy. By immunoelectron microscopy, the antibody was demonstrated to react with the GP-specific structure, germinal granules, in cleaving embryos, and with their derivatives in the germ line cells of embryos at stages extending from gastrula to feeding tadpole. It also reacted with mitochondria not only in the GP and the germ line cells but also in somatic cells, and with myofibrils in muscle cells. By Western blotting, the antibody was shown to react with several bands of Mr 42–69 ± 10
18.
Adenine depurination and inactivation of plant ribosomes by an antiviral protein of Mirabilis jalapa (MAP) 总被引:4,自引:0,他引:4
Jiro Kataoka Noriyuki Habuka Masashi Miyano Chikara Masuta Akira Koiwai 《Plant molecular biology》1992,20(6):1111-1119
Mirabilis antiviral protein (MAP) is a single-chain ribosome-inactivating protein (RIP) isolated from Mirabilis jalapa L. It depurinates the 28S-like rRNAs of prokaryotes and eukaryotes. A specific modification in the 25S rRNA of M. jalapa was found to occur during isolation of ribosomes by polyacrylamide/agarose composite gel electrophoresis. Primer extension analysis revealed the modification site to be at the adenine residue corresponding to A4324 in rat 28S rRNA. The amount of endogenous MAP seemed to be sufficient to inactivate most of the homologous ribosomes. The adenine of wheat ribosomes was also found to be removed to some extent by an endogenous RIP (tritin). However, the amount of endogenous tritin seemed to be insufficient for quantitative depurination of the homologous ribosomes.Endogenous MAP could shut down the protein synthesis of its own cells when it spreads into the cytoplasm through breaks of the cells. Therefore, we speculate that MAP is a defensive agent to induce viral resistance through the suicide of its own cells. 相似文献
19.
20.
Role of salicylic acid glucosyltransferase in balancing growth and defence for optimum plant fitness