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1.
Tomohiro Hamasaki Takahiro Matsumoto Naoya Sakamoto Akiko Shimahara Shiori Kato Ayumi Yoshitake Ayumi Utsunomiya Hisayoshi Yurimoto Esteban C. Gabazza Tadaaki Ohgi 《Nucleic acids research》2013,41(12):e126
Radioisotopes and fluorescent compounds are frequently used for RNA labeling but are unsuitable for clinical studies of RNA drugs because of the risk from radiation exposure or the nonequivalence arising from covalently attached fluorophores. Here, we report a practical phosphoramidite solid-phase synthesis of 18O-labeled RNA that avoids these disadvantages, and we demonstrate its application to quantification and imaging. The synthesis involves the introduction of a nonbridging 18O atom into the phosphate group during the oxidation step of the synthetic cycle by using 18O water as the oxygen donor. The 18O label in the RNA was stable at pH 3–8.5, while the physicochemical and biological properties of labeled and unlabeled short interfering RNA were indistinguishable by circular dichroism, melting temperature and RNA-interference activity. The 18O/16O ratio as measured by isotope ratio mass spectrometry increased linearly with the concentration of 18O-labeled RNA, and this technique was used to determine the blood concentration of 18O-labeled RNA after administration to mice. 18O-labeled RNA transfected into human A549 cells was visualized by isotope microscopy. The RNA was observed in foci in the cytoplasm around the nucleus, presumably corresponding to endosomes. These methodologies may be useful for kinetic and cellular-localization studies of RNA in basic and pharmaceutical studies. 相似文献
2.
N Imanishi T Nakayama M Asano K Yatsunami K Tomita A Ichikawa 《Biochimica et biophysica acta》1987,928(2):227-234
Dexamethasone at a concentration as low as 10 nM significantly increased both the histamine content and histidine decarboxylase activity of cultured mastocytoma P-815 cells. Both effects were clearly seen using several glucocorticoids, which were as effective as dexamethasone. In contrast to that of histamine, the serotonin level of mastocytoma P-815 cells was decreased by treatment with dexamethasone. The dexamethasone-induced increases in histamine content and histidine decarboxylase activity were completely suppressed by the addition of cycloheximide and actinomycin D. Mastocytoma P-815 cells were found to possess binding sites for [3H]dexamethasone in the cytosol (Kd = 15.7 nM) and the nuclei (Kd = 1.26 nM). These results show that glucocorticoids significantly stimulate de novo synthesis of histidine decarboxylase. 相似文献
3.
J Imanishi T Oku Y Cho S Inagawa A Tanaka W Kuwayama 《Comptes rendus des séances de la Société de biologie et de ses filiales》1985,179(3):414-419
In the present study we show that peritoneal macrophages obtained from the mice treated with the immunoactive peptides inhibit the multiplication of Herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), but not that of vesicular stomatitis virus (VSV), and that the intraperitoneal administration of the peptides suppresses the infection with HSV-1 in mice. 相似文献
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T Uchiyama T Tadakuma K Imanishi M Araake S Saito X J Yan H Fujikawa H Igarashi N Yamaura 《Journal of immunology (Baltimore, Md. : 1950)》1989,143(10):3175-3182
Toxic shock syndrome toxin-1 (TSST-1)-binding structures present on murine lymphoid tissues were investigated by using 125I-TSST-1. T-depleted C57BL/6 spleen cells incubated with TSST-1 for 3 h at 0 degree C were mitogenic to splenic T cells, indicating that the former cells bind and present TSST-1 to T cells. TSST-1-binding activity was observed in C57BL/6 splenic B cells and L cells transfected with I-Ab genes, but not in splenic T cells and control L cells. Scatchard plot analysis showed that these B cells and transfectants bound TSST-1 with similar binding affinity. SDS-PAGE analysis showed that lysates of C57BL/6 spleen cells and the I-Ab-positive transfectants contain a single band which bound TSST-1 and comigrated with I-Ab heterodimers. TSST-1-binding activity observed clearly in C57BL/6. BALB/c, and C3H/HeN spleen cells and L cells transfected with I-Ab or I-Ak genes was not reduced by paraformaldehyde fixation. Binding of 125I-TSST-1 to the three spleen cells was markedly reduced by anti-I-A antibodies, but not by anti-I-E antibodies. C57BL/6, C3H/HeN, and (C3H/HeN x C57BL/6) F1 T cells were activated by TSST-1 to proliferate and produce IL-2 in the presence of FT6.2 cells, LT1-30-3 cells and either of them, respectively, but not in the presence of control L cells. These results indicate that I-A molecules function as the structures via that accessory cells directly bind TSST-1 on the cell surface and present a triggering signal of TSST-1 to T cells. 相似文献
6.
M Kita Y Ohmoto Y Hirai N Yamaguchi J Imanishi 《Comptes rendus des séances de la Société de biologie et de ses filiales》1990,184(5-6):380-384
Mycoplasma shows a variety of effects on immune system, including the activation of macrophage, the increase in T cell cytotoxicity, and the enhancement of the proliferation and maturation of B cells, etc. As it is well known that many cytokines regulate the immune system, it is interesting to examine whether or not human peripheral blood mononuclear cells (PBMC) produce interleukin (IL) in response to mycoplasmas. In the present study, human PMBC were incubated with 7 species of mycoplasmas for 48 hours, and IL-1 beta, IL-2 and IL-6 activities in the supernatants were determined by ELISA. All the species of mycoplasmas were able to induce IL-1 beta and IL-6, although IL-2 was induced only by M. pneumoniae. These results suggest that the influence of mycoplasma infection on immune system may be partly due to the interleukins induced by mycoplasmas. 相似文献
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8.
Sakaba Tomoka Soejima Akiko Fujii Shinji Ikeda Hajime Iwasaki Takaya Saito Hiroaki Suyama Yoshihisa Matsuo Ayumi Kozhevnikov Andrey E. Kozhevnikova Zoya V. Wang Hongfeng Wang Siqi Pak Jae-Hong Fujii Noriyuki 《Journal of plant research》2023,136(4):437-452
Journal of Plant Research - A group of temperate grassland plant species termed the “Mansen elements” occurs in Japan and is widely distributed in the grasslands of continental East... 相似文献
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10.
Comparative studies on the inhibitory effects of interferon on various strains of herpes simplex viruses in vitro 总被引:2,自引:0,他引:2
The sensitivities to human leukocyte interferon of 10 strains of Herpes simplex virus (HSV) type 1 and 3 strains of type 2 were compared. All the strains were sensitive to interferon, although their sensitivities were less than that of vesicular stomatitis virus (VSV). There was no significant difference in the sensitivities to interferon of HSV type 1 and type-2 or among different strains of a given type of HSV. Nor was there any difference between the sensitivities of 5-iodo-2'-deoxyuridine (IDU)-sensitive and resistant strains isolated from the same patients. These results suggest that interferon should be useful in therapy of HSV infection. 相似文献