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61.
62.
The effect of dimethylnitrosamine on the nucleosomal structure of mouse liver chromatin was studied. After a single oral dose of dimethylnitrosamine (2–75 mg/kg body weight 45 min before sacrifice) liver nuclei were isolated and incubated with micrococcus nuclease. Nucleosomes were separated on sucrose density gradients. There were no differences in nucleosomal sedimentation velocities between preparations from control and dimethylnitrosamine treated animals. The supernatant obtained after centrifugation of the lysed nuclei (2 min at 4,000 g
av) and nucleosomal peak fractions were used for isolation of DNA. DNA was heat denatured in 7 M urea or formamide. After electrophoresis on polyacrylamide gels areas under mononucleosomal DNA and smaller fragments were measured and compared with the total DNA area. The increase in DNA fragmentation was dimethylnitrosamine dose response dependent. When expressed as per cent of controls it amounted to 106% for 2 mg; 115% for 10 mg; 127% for 25 mg; 164% for 75 mg dimethylnitrosamine/kg body weight. A good correlation between mobility and log of chain length of
174 RF DNA-Hae III digest was obtained in nondenaturing 5% polyacrylamide gels and denaturing non-aqueous formamide polyacrylamide gels but not in 12% polyacrylamide gels containing 7 M urea. DNA of mononucleosomal peak fractions contained 200 and that of dinucleosomal peak fractions 400 nucleotides. Fragmentation of DNA was closely related to in vivo dimethylnitrosamine treatment but was not detected in measurements of protein-DNA complexes in the chromatin. It was disclosed on denaturation of DNA followed by polyacrylamide gel electrophoresis.Abbreviations DMN
dimethylnitrosamine
- SDS
sodium dodecyl sulfate
The work was supported by Grant Number 1 R0 1 CA26642-01, awarded to A.v.d.D. by the National Cancer Institute, DHEW 相似文献
63.
目的 研究预防非典型肺炎方剂对小鼠外周血液淋巴细胞 DNA的损伤作用。方法 应用单细胞凝胶电泳方法测定 6个预防非典型肺炎方剂体内给药对小鼠外周血液淋巴细胞 DNA损伤的作用。结果 ig给予预防非典型肺炎方剂 、 、 (1/ 3临床等效剂量、临床等效剂量、3倍临床等效剂量× 3 d)对小鼠外周血液淋巴细胞 DNA有明显的损伤作用 ,并呈剂量依赖关系。预防非典型肺炎方剂 、 、 (1/ 3临床等效剂量、临床等效剂量、3倍临床等效剂量× 3 d)对小鼠外周血液淋巴细胞 DNA没有明显的损伤作用。结论 预防非典型肺炎方剂 、 、 具有潜在的遗传毒性 相似文献
64.
65.
左氧氟沙星滴眼液及眼用凝胶的研究概述 总被引:1,自引:0,他引:1
目的 对近年来左氧氟沙星在眼用制剂方面的研究作一综述。方法 参考及整理有关文献。结果 总结并归纳了左氧氯沙星滴眼及眼用凝胶的研究进展。结论:左氧氟沙星缓释型眼用制剂越来越受到重视和发展,并具有广泛的临床应用前景。 相似文献
66.
双向电泳技术已广泛用于蛋白质组学研究,该技术除了在农业和基础医学领域中有着广泛的应用外,还在疫苗的研究、开发和生产中具有重要的应用前景.此文就双向电泳技术在疫苗抗原及疫苗候选菌株的筛选、基因工程疫苗的研究、抗原呈递及免疫应答机制的研究和疫苗生产过程中批间一致性分析等方面的应用进行了综述. 相似文献
67.
目的 为了研究菲啰啉对2种氧化剂和抗癌药多柔比星诱发细胞DNA损伤的影响, 并初步探讨其损伤机制。方法 用不同浓度菲啰啉预处理CHL细胞30 min, 再分别加入3种不同染毒受试物,共同培养一定时间(0.3 mmol·L-1重铬酸钾:105 min; 0.5 μmol·L-1多柔比星:5 min; 0.4 mmol·L-1过氧化氢(H2O2):25 min)后, 用碱性单细胞凝胶电泳方法(ASCGE)测定DNA链断裂情况, 并同时以菲啰啉与二甲亚砜(DMSO,0.33 mol·L-1)比较对H2O2致DNA损伤中·OH的产生和清除。结果 3种染毒受试物均可明显引起CHL细胞DNA链断裂;而当3 μmol·L-1菲啰啉预处理后, 可使重铬酸钾、H2O2所致DNA迁移长度和细胞拖尾率明显降低, 并超过DMSO降低H2O2的损伤作用, 当菲啰林浓度升至12 μmol·L-1时, 可完全消除这两种因素所致的DNA链断裂损伤;10 μmol·L-1菲啰啉可抑制多柔比星所致DNA损伤, 但浓度直至60 μmol·L-1仍不能完全消除多柔比星的损伤作用。结论 菲啰啉对2种氧化剂和多柔比星所致DNA损伤均有不同程度的防护作用,同时提示重铬酸钾和H2O2所致的DNA损伤主要与需过渡金属离子参与的·OH产生有关, 而多柔比星所致损伤仅部分与此有关。 相似文献
68.
The transdermal delivery of buspirone hydrochloride across hairless mouse skin and the combined effect of iontophoresis and terpene enhancers were evaluated in vitro using Franz diffusion cells. Iontophoretic delivery was optimized by evaluating the effect of drug concentration, current density, and pH of the vehicle solution. Increasing the current density from 0.05 to 0.1 mA/cm2 resulted in doubling of the iontophoretic flux of buspirone hydrochloride, while increasing drug concentration from 1% to 2% had no effect on flux. Using phosphate buffer to adjust the pH of the drug solution decreased the buspirone hydrochloride iontophoretic flux relative to water solutions. Incorporating buspirone hydrochloride into ethanol:water (50:50 vol/vol) based gel formulations using carboxymethylcellulose and hydroxypropylmethylcellulose had no effect on iontophoretic delivery. Incorporation of three terpene enhancers (menthol, cineole, and terpineol) into the gel and when combined with iontophoresis it was possible to deliver 10 mg/cm2/day of buspirone hydrochloride. 相似文献
69.
Abstract: We used a N‐biotinylated peptide analog of the C‐terminal domain of the tumor suppressor protein, p21cip1/waf1 to elucidate peptide/protein interacting partners. The C‐terminal domain of p21cip1/waf1 protein spanning 141–160 amino acid residues is known to bind PCNA and this interaction is important in many biological processes including cell‐cycle control. This C‐terminal 20‐mer efficiently extracts PCNA in the presence of a variety of N‐ or C‐terminally attached affinity tags. Using difference silver stained 2D gels combined with in‐gel tryptic digests, we identified the difference spots using MALDI‐TOF mass spectrometry‐based peptide mass fingerprinting followed by a database search using profound against NCBIs human nonredundant protein sequence data bank. Identified spots include the p48 subunit of chromatin assembly factor‐1, the heat shock 70 protein analog BiP, calmodulin, nucleolin and a spot similar in size to dimeric PCNA. In contrast, microcapillary ion‐trap LC‐MS/MS analysis of a tryptic digest of entire affinity extracts derived from both control and experimental runs followed by database searches using sequest confirmed the presence of most of the above proteins. This strategy also identified hnRNPA1, HPSP90α, HSP40 and T‐complex protein 1, a protein similar to prothymosin, and a possible allelic variant of the p21cip1/waf1 protein. The use of N‐biotinylated peptide derived from the C‐terminal domain of p21cip1/waf1 protein in proteomic analysis exemplified here suggests that peptides obtained from intracellular functional screens could also potentially serve as efficient baits to discover new drug targets. 相似文献
70.