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Imtiaj Hasan Shigeki Sugawara Yuki Fujii Yasuhiro Koide Daiki Terada Naoya Iimura Toshiyuki Fujiwara Keisuke G. Takahashi Nobuhiko Kojima Sultana Rajia Sarkar M. A. Kawsar Robert A. Kanaly Hideho Uchiyama Masahiro Hosono Yukiko Ogawa Hideaki Fujita Jiharu Hamako Taei Matsui Yasuhiro Ozeki 《Marine drugs》2015,13(12):7377-7389
MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface. 相似文献
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Hamako Sasamoto Yohichi Wakita Shinso Yokota Nobuo Yoshizawa 《Journal of Forest Research》2000,5(4):265-270
An improved method for selection of large electro-fused protoplasts ofPopulus alba by a micromanipulator was developed. The conditions for electric cell fusion treatment were optimized. For the best result,
protoplasts with a cell density of 5 × 105/mL were treated with an alternate current (1 MHz, 200 V/cm) and pulsed with a direct current (2 kV/cm) for 100μs in 2.5 mM CaCl2 and 0.55 M mannitol. The electo-fused protoplasts were cultured in NH4NO3-free Murashige and Skoog’s medium containing 0.6 M of mannitol, 0.09 M sucrose, 1μM of 2,4-dichlorophenoxyacetic acid and 0.1μM of benzyladenine, the same medium used for protoplast culture, but at a very low cell density of 5–10 × 102/mL in a well of a 96-well culture plate. When cell aggregates derived from individual fused protoplasts were transferred
to fresh medium with 0, 0.3 or 0.6 M mannitol, large colonies developed. In the shoot differentiation medium, the reaction
of the calluses derived from large fused protoplasts towards the growth regulators differed from the non-fused ones. In medium
containing 1μM each of naphthalene acetic acid andN-(2-chloro-4-pyridyl)-N′-phenylurea, growth of callus from electro-fused ones was not reduced by much compared to the control, but shoot differentiation
was inhibited. Gibberellic acid (0.1–10μM) was beneficial to shoot regeneration; however, irregularly shaped leaves appeared at high gibberellic acid concentrations.
Shoots regenerated were rooted in Murashige and Skoog’s medium containing 4μM of indole-3-butyric acid. Some plantlets obtained had a varied morphology. Based on the characteristics of growth, some
cells derived from electro-fused protoplasts appear to be physiologically different from the non-fused one. 相似文献
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Shinjiro Ogita Hisako Ishikawa Takafumi Kubo Hamako Sasamoto 《Journal of Wood Science》1999,45(2):87-91
Embryogenic cells (ECs) of sugi (Cryptomeria japonica D. Don) were induced from immature and mature zygotic embryos cultured on different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D)-containing modified Campbell and Durzan medium. The rate of induction of ECs varied depending on the stage of embryos collected. The highest percentage of induction (35%) was obtained with immature zygotic embryos collected July 18 and July 30, 1997, when 1 M of 2,4-D was added to the induction medium. The ECs easily proliferated when subcultured in a medium of the same composition as the induction medium within 3 weeks. Morphological characteristics of nonembryogenic cells and embryogenic cells of different developmental stages were studied under an inverted fluorescence microscope.Part of this work was presented at the 109th annual Japanese Forestry Society meeting at Tochigi, April 1998 相似文献
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Two embryogenic cell lines characterized by different morphology and color, white and red, were obtained from an immature
zygotic embryo of Japanese larch (Larix leptolepis Gord.). Mature somatic embryos with cotyledons and regenerated plants were obtained from both cell lines. However optimal
concentration of abscisic acid (ABA) for maturation varied depending on morphology of ECs. From the white ECs which intermingled
with somatic embryos of very early stage, mature somatic embryos were induced on maturation medium containing 50 μM of ABA.
On the other hand, mature somatic embryos with cotyledons were observed from the red ECs which consisted of somatic embryos
of more developed stage on hormone-free medium or 0.1 μM ABA containing-medium. 相似文献
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