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11.
G L Whitson W L Carrier A A Francis C C Shih S Georghiou J D Regan 《Cell and tissue kinetics》1986,19(1):39-47
A number of physical and chemical agents in the environment have been studied for their ability to induce or alter DNA repair mechanisms in human cells. We have investigated the effects of 60 Hz, 1000 V/cm electric fields on DNA repair in normal human fibroblasts in vitro. An examination was done on the ability of electric fields suspected to cause damage which could be repaired by thymine dimer excision and measurable by the bromodeoxyuridine photolysis assay. The thymine dimer assay with enzyme-sensitive site analysis was used to measure the cells' capacity for removing ultraviolet light (u.v.)-induced pyrimidine dimers; during exposure to electric field 24 hr before u.v. irradiation; 24 hr after u.v. irradiation; and up to 48 hr continuously after u.v. irradiation. Cell growth and cell survival following electric field exposure were also studied. Within the limits of these experiments, it was found that exposure to such electric fields did not alter cell growth or survival, and no DNA repair or alteration in DNA excision repair capacity was observed as compared with unexposed control cultures. 相似文献
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Reactivity of a sulfhydryl group of the ras oncogene product p21 modulated by GTP binding 总被引:4,自引:0,他引:4
S Hattori T Yamashita T D Copeland S Oroszlan T Y Shih 《The Journal of biological chemistry》1986,261(31):14582-14586
We have studied the sensitivity of sulfhydryl groups of a highly purified p21 protein of the v-rasH oncogene to a thiol-specific reagent, N-ethylmaleimide (NEM). Approximately 70% of GTP binding and autokinase activities of p21 were inactivated by NEM, and excessive amounts of GTP or GDP protected p21 activities. Thiol titration revealed the presence of one fast reactive cysteine residue, the susceptibility of which is modulated by GTP binding. A total of 4 and 6 residues, respectively, became titratable upon denaturation and reduction, suggesting the presence of a disulfide bond. This GTP-modulated sulfhydryl group was identified as Cys-80 in the following tryptic peptide sequence: NH2-Thr-Gly-Glu-Gly-Phe-Leu-Cys-Val-Phe-Ala-Ile-Asn-Asn-Thr-Lys-COOH. This is based on the comparative tryptic peptide mapping of [14C]NEM-modified p21 in the presence and absence of GTP. The GTP-modulated peptide co-chromatographed with a synthetic peptide of the predicted sequence. Amino acid analysis of the purified [14C]NEM-modified peptide from tryptic digests of p21 also confirmed its identity. This region of p21 shares an extensive sequence homology with various G-proteins and appears to be in the vicinity of the GTP-binding domain of these proteins. 相似文献
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A simple and low-cost poultry waste digester (PWD) was constructed to treat the waste from 4000 caged laying hens on University Research Unit No. 2 at North Carolina State University. The system was built basically of a plastic lining with insulation, a heating system, a hot-water tank, and other metering equipment. It was operated at 50 degrees C and pH 7.5-8.0. The initiation of methane production was achieved using the indigenous microflora in the poultry waste. At an optimal loading rate (7.5 kg volatile solids/m(3) day), the PWD produced biogas (55% methane) at a rate of 4.0 m(3)/m(3) day. The PWD was biologically stable and able to tolerate temporary overloads and shutdowns. A higher loading rate failed to maintain a high gas production rate and caused drops in methane content and pH value. Under optimal conditions, a positive energy balance was demonstrated with a net surplus of 50.6% of the gross energy. For methane production, the PWD system was proved to be technically feasible. The simple design and inexpensive materials used for this model could significantly reduce the cost of digestion compared to more conventional systems. More studies are needed to determine the durability, the required maintenance of the system, and the most economical method of biogas and solid residue utilization. 相似文献
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R L Truitt C Y Shih A V Lefever L D Tempelis M Andreani M M Bortin 《Journal of immunology (Baltimore, Md. : 1950)》1983,131(4):2050-2058
We have reported that immunization of H-2k mice with lymphoid cells from various allogeneic strains induced a population of cells that could eliminate first-passage spontaneous AKR leukemia from the spleens of immuno-suppressed AKR (H-2k) hosts. In the present study, we examined the nature of the cells responsible for this graft-vs-leukemia (GVL) reaction and compared them to cytolytic cells detected in vitro. Spleen cells from alloimmunized CBA/J (H-2k) mice were selectively depleted of various subpopulations by treatment with antibody and complement (C), then tested in vivo for GVL reactivity. Cell suspensions depleted of Thy-1.2+, Lyt-1+, or Lyt-2+ lymphocytes had no significant GVL reactivity, whereas suspensions depleted of NK-1.2+ cells retained GVL reactivity. The GVL-reactive cells persisted in H-2-compatible donor mice for up to 56 days. Lyt-1+2+ lymphocytes that were cytotoxic for cultured AKR leukemia cells in vitro could be detected in the spleens of alloimmunized H-2-compatible mice after expansion of the cells in T cell growth factor. Using quantitative limiting dilution cytotoxicity assays, we found that the frequency of leukemia-reactive cytotoxic lymphocytes (CL) in the spleen showed a direct correlation with the GVL efficacy of the cells in vivo. Alloimmunization was essential for induction of the GVL-reactive cell population. CL in alloimmunized mice consisted of heterogeneous cytotoxic specificities; i.e., some CL were leukemia-specific, others lysed only nonleukemic AKR target cells, and a third group mediated killing of both leukemic and nonleukemic target cells. The CL appeared to be H-2 restricted and specific for non-H-2 antigens shared by the AKR leukemia and the alloimmunizing cells. 相似文献
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Detachment and incubation of Elodea leaves promoted callose synthesis in all cells, especially in epidermal pits and in sieve tubes. Phloem was detected in the midrib by fluorescent staining of callose induced to form on sieve plates. In EM views of mature sieve elements nucleus and tonoplast were lacking, mictoplasm replaced cytoplasm, mitochondria were fewer in number, and large plastids contained crystalline inclusion bodies. Slime was present as compact aggregates and as individual fibrils in mictoplasm and sieve pores. Deposition of callose is considered in relation to the blockage concept of callose function. 相似文献
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