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71.
Efficient induction of antitumor cytoxic T lymphocytes from a healthy donor using HLA-A2-restricted MAGE-3 peptide in vitro 总被引:1,自引:0,他引:1
F. Tanaka Tatsuo Fujie Hiroki Go Kinya Baba Masaki Mori Kazutoh Takesako Tsuyoshi Akiyoshi 《Cancer immunology, immunotherapy : CII》1997,44(1):21-26
The antigenic peptides encoded by tumor-rejection antigen genes, MAGE-1 and -3, have been identified, and various methods have been utilized for the in vitro induction of MAGE-specific, cytotoxic
T lymphocytes (CTL) from peripheral blood mononuclear cells (PBMC) using synthetic peptides. However, all of these methods
are technically demanding and thus have a relatively limited usefulness. We herein report a simple and efficient method for
the in vitro induction of specific CTL by using the HLA-A2-restricted MAGE-3 peptide from the PBMC of a healthy donor. CTL
responses could thus be efficiently induced from unseparated PBMC by stimulation with freshly isolated, peptide-pulsed PBMC
as antigen-presenting cells and by using interleukin-7 and keyhole limpet hemocyanin for the primary culture. The induced
CTL could thus recognize and lyse not only HLA-A2 target cells pulsed with the peptide but also HLA-A2 tumor cells expressing
MAGE-3, in an HLA-class-I-restricted manner. This simple method may, therefore, become a useful tool for investigating the
potential peptides for tumor antigens as well as for developing various immunotherapeutic approaches for human malignant tumors.
Received: 15 October 1996 / Accepted: 6 December 1996 相似文献
72.
The promoter of an anther tapetum-specific gene,Osg6B, was fused to a-glucuronidase (GUS) gene and introduced into rice byAgrobacterium-mediated gene transfer. Fluorometric and histochemical GUS assay showed that GUS was expressed exclusively within the tapetum of anthers from the uninucleate microspore stage (7 days before anthesis) to the tricellular pollen stage (3 days before anthesis). This is the first demonstration of an anther-specific promoter directing tapetum-specific expression in rice.Abbreviations
GUS
ßGlucuronidase 相似文献
73.
Changes in Levels of Purine and Pyrimidine Nucleotides During Acute Hypoxia and Recovery in Neonatal Rat Brain 总被引:6,自引:2,他引:4
Neonatal rat brains were examined for changes in levels of ATP, ADP, AMP, cyclic AMP, GTP, GDP, UTP, UDP, UMP, and CTP during exposure to 100% nitrogen for 20 min and subsequent recovery in air. During hypoxia, ATP, GTP, UTP, and CTP levels and the GTP/GDP ratio decreased to 38, 50, 26, 21, and 21%, respectively, of control levels. No significant change in cyclic AMP level was observed. The decrease in the total uridine nucleotide pool during hypoxia was markedly greater (to 53% of control levels) than that in the total adenine nucleotide pool (to 92% of control levels). During recovery, ATP and GTP levels were rapidly and almost completely restored. On the other hand, CTP levels returned slowly to control values after a 2-h recovery period. Restoration of the UTP level was slow and incomplete (87% of the control value even after a 3-h recovery period). The GTP/GDP ratio also did not return to normal. These data suggest that hypoxic insult to the neonate may have an effect on the synthesis of nucleotidyl sugars, phospholipids, and proteins in the brain, resulting in significant problems with developmental processes of the brain. The present study also showed that the delayed restorations of the UTP level and the GTP/GDP ratio were not seen in the brains of adult rats subjected to acute severe hypoxic insult. 相似文献
74.
Purification and Characterization of Benzonitrilases from Arthrobacter sp. Strain J-1 总被引:9,自引:5,他引:4 下载免费PDF全文
Amal Kumar Bandyopadhyay Toru Nagasawa Yasuhisa Asano Kinya Fujishiro Yoshiki Tani Hideaki Yamada 《Applied microbiology》1986,51(2):302-306
We found two kinds of benzonitrilases, designated benzonitrilases A and B, in a cell extract of Arthrobacter sp. strain J-1 grown on benzonitrile as a sole carbon and nitrogen source. Benzonitrilases A and B were purified approximately 409-fold and 38-fold, respectively. Purified benzonitrilase A appeared to be homogeneous according to the criteria of polyacrylamide gel electrophoresis. Both the enzymes hydrolyzed benzonitrile to benzoic acid and ammonia without forming benzamide as an intermediate. The molecular weights of benzonitrilases A and B were found to be 30,000 and 23,000, respectively. The subunit molecular weight of benzonitrilase A was the same as its molecular weight. The isoelectric points of benzonitrilases A and B were 4.95 and 4.80, respectively. The optimum temperature and pH, respectively, for benzonitrilase A were 40°C and 8.5, and those for benzonitrilase B were 30°C and 7.5. The Km values for benzonitrilases A and B were 6.7 mM and 4.5 mM, respectively. Both the enzymes degraded p-tolunitrile, 4-cyanopyridine, and p-chlorobenzonitrile, but they did not attack aliphatic nitriles or amides. Both the enzymes were inhibited by thiol reagents. 相似文献
75.
Nocardomycolic acids from Nocardia rubra were fully separated and characterized by a combination of argentation thin-layer chromatography and gas chromatography — mass spectrometry (GCMS). The occurrence of 20 or more different molecular species of mycolic acids was demonstrated. GCMS analysis of each subclass of mycolic acids after separation on AgNO3 thin-layer chromatography revealed that in general the major species consisted of the even-carbon mycolic acids ranging from C38 to C52. However, the most abundant species differed by the subclasses; C44 being in saturated, C46 in monoenoic and C46 in dienoic mycolic acids, respectively. All these acids were shown to possess C12 or C14 alkyl branch at 2 position, while double bonds were located in longer straight chain alkyl unit.By using this method, distinctive changes in mycolic acid composition by growth temperature were observed. The ratios of saturated, monoenoic to dienoic mycolic acids in a mixture of certain carbon numbered mycolic acids varied greatly, according to the shift of growth temperature. The mass fragmentographic analysis, monitoring M-15 ions derived from the loss of methyl group from the molecular ions showed the lower temperature (15°C) grown cells contained more unsaturated (especially dienoic) mycolic acids, while the higher temperature (40°C) grown cells contained more saturated mycolic acids in both extractable and cell-wall bound lipids. These changes in mycolic acid composition occurred shortly after shifting up the growth temperature from 20°C to 43°C at a logarithmic stage of the bacterial growth. 相似文献
76.
Kinya Kuriyama Etsuo Kurihara Yoshihisa Ito Yukio Yoneda 《Journal of neurochemistry》1980,35(2):343-348
The effect of intrastriatal microinjection of kainic acid (KA) on specific binding of [3H]muscimol to the particulate fractions obtained from corpus striatum (CS), globus pallidus (GP), substantia nigra (SN), and cerebral cortex (CC) was examined. Seven days after the unilateral intrastriatal microinjection of KA, the amount of specifically bound [3H]muscimol was significantly increased at the injected site, whereas no significant alteration of [3H]muscimol binding was found in GP, SN, or CC. Scatchard analysis of striatal binding revealed that microinjection of KA significantly increased the affinity (KD) of GABA receptors on the injected (lesioned) side of the CS without affecting the total number of binding sites (Bmax) therein. This significant increase in [3H]muscimol binding, however, was eliminated by pretreating particulate fractions from the CS with Triton X-100, a non-ionic detergent. No statistically significant difference in amounts of [3H]muscimol binding was detected when the preparations from the KA-treated and non-treated CS were preincubated with 0.05% Triton X-100, respectively. Scatchard analysis using CS preparations treated with 0.05% Triton X-100 revealed that the affinity of the GABA receptor was increased by treatment with Triton X-100, while the total number of binding sites (Bmax) was unchanged by this treatment. These results suggest that neuronal degeneration produced by KA in vivo and pretreatment of particulate preparations with Triton X-100 in vitro may increase the amount of specifically bound [3H]muscimol to CS preparations by a similar molecular mechanism. 相似文献
77.
Kyoji Nakajima Ikuo Tooyama Kinya Kuriyama Hiroshi Kimura 《Neurochemical research》1996,21(2):211-215
Immunohistochemical localization of GABAB-receptors was demonstrated in the rat gastrointestinal tract using a monoclonal antibody (GB-1) raised against the purified
GABAB-receptor. Immunoreactive staining for GABAB-receptors was found in some populations of endocrine, muscular and neuronal components in the stomach and gut wall. Positive
mucosal epithelial, probably endocrine, cells were distributed throughout the stomach and intestine. Double immunostaining
indicated that such positive cells for GABAB-receptors often co-possessed serotonin in the small intestine but not in the gastric body. In the muscular layer of the digestive
canal, positive staining was seen as dotty granules punctuated on the surface of muscle fibers. In the enteric nervous system,
positive neuronal somata were found in both submucosal and myenteric ganglia throught the entire canal extending from the
stomach to the rectum. This is the first report to visualize the cellular localization of GABAB-receptors in the gastrointestinal system of the rat, and should provide a fundamental basis for future studies on gastrointestinal
functions regulated by GABAB-receptors.
Special issue dedicated to Dr. Kinya Kuriyama. 相似文献
78.
Okamoto K Ocker M Neureiter D Dietze O Zopf S Hahn EG Herold C 《Journal of cellular and molecular medicine》2007,11(2):349-361
Gemcitabine has been shown to ameliorate disease related symptoms and to prolong overall survival in pancreatic cancer.Yet, resistance to Gemcitabine is commonly observed in this tumour entity and has been linked to increased expression of anti-apoptotic bcl-2. We therefore investigated if and to what extend silencing of bcl-2 by specific siRNAs (siBCL2) might enhance Gemcitabine effects in human pancreatic carcinoma cells. siBCL2 was transfected into the pancreatic cancer cell line YAP C alone and 72 hrs before co-incubation with different concentrations of Gemcitabine. Total protein and RNA were extracted for Western-blot analysis and quantitative polymerase chain reaction. Pancreatic cancer xenografts in male nude mice were treated intraperitoneally with siBCL2 alone, Gemcitabine and control siRNA or Gemcitabine and siBCL2 for 21 days. Combination of both methods lead to a synergistic induction of apoptosis at otherwise ineffective concentrations of Gemcitabine. Tumour growth suppression was also potentiated by the combined treatment with siBCL2 and Gemcitabine in vivo and lead to increased TUNEL positivity. In contrast, non-transformed human foreskin fibroblasts showed only minor responses to this treatment. Our results demonstrate that siRNA-mediated silencing of anti-apoptotic bcl-2 enhances chemotherapy sensitivity in human pancreatic cancer cells in vitro and might lead to improved therapy responses in advanced stages of this disease. 相似文献
79.
80.
T Kamiya A Takano Y Matsuzuka N Kusaba M Ikeguchi K Takagaki K Kondo 《Bioscience, biotechnology, and biochemistry》2012,76(8):1511-1517
In Japan, kudzu is a familiar plant, well-known as an ingredient in the Japanese-style confections kudzu-kiri and kudzu-mochi. In this study, we focused on the flower of kudzu (Pueraria thomsonii) and conducted a clinical trial to investigate the effects of Pueraria thomsonii flower extract (PFE) on obesity using obese Japanese males and females (BMI ≥ 25 kg/m(2)). Eighty-one obese subjects were randomly divided into three groups and consumed test food containing 300 mg of PFE, 200 mg of PFE, and a placebo over 12 weeks. The results indicate that PFE intake reduces BMI and decreases, the visceral fat area, but not the subcutaneous fat area. In addition, the decrease in visceral fat area showed no sexual dimorphism. Consequently, we propose that PFE intake expresses its BMI reduction effects via a decrease in visceral fat area. 相似文献