DNA repair and replication were examined in diploid human fibroblasts after treatment with (±)-r-7,t-8-dihydroxy-t-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE-I). Unscheduled DNA synthesis exhibited a linear response to BPDE-I concentrations up to 1.5 μM and a saturation plateau after higher concentrations. Maximal unscheduled DNA synthesis was observed in the first hour after treatment with synthesis diminishing progressively thereafter. Half-maximal unscheduled DNA synthesis was seen within 4–6 h after treatment with 0.7 μM BPDE-I. DNA replication was inhibited by BPDE-I in a dose- and time-dependent fashion. The mechanisms of this inhibition were characterized by velocity sedimentation of pulse-labeled nascent DNA in alkaline sucrose gradients. Very low concentrations of BPDE-I (0.03 and 0.07 μM) were found to inhibit replicon initiation by up to 50% within 30–60 min after treatment. Recovery of initiation following these low concentrations was evident within 3 h after treatment. Higher concentrations of carcinogen inhibited DNA synthesis in active replicons. This effect was manifested by a reduction in incorporation of precursor into replication intermediates of greater than 1·107 Da with the concurrent production of abnormally small nascent DNA. When viewed 45 min after treatment with 0.17 μM BPDE-I the combination of these two effects partially masked the inhibition of replicon initiation. However, even after treatment with 0.33 μM BPDE-I an effect on initiation was evident. These results reveal a pattern of response to BPDE-I that is quite similar to that produced by 254 nm radiation. 相似文献
400 MHz 1H NMR of ferric low-spin cytochrome P-450scc purified from bovine adrenal cortex was measured for the first time. As compared with 1H NMR spectra of low-spin P-450cam and metMb- mercaptan complexes, paramagnetic shifts of low-spin P-450scc complexes were more divergent, suggesting that there is a subtle difference in the heme environment between P-450scc and P-450cam [1]. The paramagnetic shifts of low-spin complexes of P-450scc caused by adding nitrogenous inhibitors, aminoglutethimide and metyrapone, were different from those caused by adding an intermediate, 20α-hydroxycholesterol, and a detergent, Tween 20 [2]. The paramagnetic shifts of the metMb-mercaptan complexes were convergent compared with those of ferric low-spin P-450scc and P-450cam, suggesting that the electronic character and/or the conformation of the internal thiolate ligand in P-450scc and P-450cam are different from those of the external thiolate ligand in metMb-thiolate complexes [3]. The paramagetic shifts of the metMb-mercaptan complexes were dependent on the electron donating factor of the alkyl group of the bound mercaptans [4].Magnetic CD(MCD) spectra of ferric low-spin P-450scc, rabbit liver P-450 complexes and metMb- mercaptan complexes were also observed at various temperatures. The temperature dependences of the Soret MCD bands for the low-spin P-450 and metMb- mercaptan complexes were decidedly less pronounced than those for the low-spin metMb-CN? or imidazole complexes, suggesting that thiolate ligands markedly influence the Soret MCD band of the ferric low-spin complexes [1]. The suggestion described in [2] implied by the 1H NMR study was reconfirmed from the temperature dependence study of the Soret MCD [2]. The temperature dependences of the Soret MCD bands for low-spin P-450 complexes having a non-nitrogenous ligand were more pronounced than for those having a nitrogenous ligand. 相似文献
Eleman-8β,12-olides containing a β-hydroxyl group at C-9 exist in equilibrium with 8β-hydroxyeleman-9,12-olides. Acetylation of 9β-hydroxyelemanolides gave a mixture of 8β-and 9β-acetyl derivatives in which the former predominates. The structures of zinaflorin II acetate, zinaflorin III and other 9β-hydroxyelemanolide acetates have been revised. The absolute stereochemistry of the δ-lactone juniperin has been established by means of an X-ray crystallographic analysis. 相似文献
The female‐produced sex pheromone of the Durra stem borer, Sesamia cretica (Lederer) (Lepidoptera: Noctuidae), had been previously characterized as a 75:25 blend of (Z)‐9‐tetradecenol (Z9‐14:OH) and (Z)‐9‐tetradecenyl acetate (Z9‐14:OAc) based on field trapping experiments. The low attraction of this blend in the field led us to further investigate the sex pheromone of this pest. Coupled gas chromatography with electroantennographic detection (GC‐EAD) analysis of female pheromone gland extracts consistently revealed three EAD‐active compounds. According to their GC retention times, mass spectra, and comparative EAG analyses with authentic standards, two of these compounds were found to be the previously reported components Z9‐14:OH and Z9‐14:OAc, whereas a third compound was identified as (Z)‐11‐hexadecenol (Z11‐16:OH). In wind tunnel experiments, the highest male responses were elicited by ratios of Z9‐14:OH, Z9‐14:OAc, and Z11‐16:OH, ranging from 90:1:9 to 90:5:5. In field tests, the 90:1:9 ratio of the blend loaded onto rubber septum dispensers was significantly more effective than single‐component, two‐component, and any other ratio of the three‐component blend. The greater effectiveness of this blend resulted in a more accurate detection of S. cretica flight activity compared with the previously reported two‐component blend. 相似文献
The fluorescent properties of 9-aminoacridine were studied in chloroplasts and phospholipid liposomes.
In energized chloroplasts it was found that the percentage of fluorescence quenching was dependent on both the 9-aminoacridine concentration and the chlorophyll concentration. On the other hand, it was independent of the osmolarity of the medium.
In phospholipid liposomes the dependence of the fluorescence quenching on the concentration of 9-aminoacridine was similar to that in chloroplasts. Moreover, the fluorescence quenching depended on the presence of charged compounds in the membrane being larger in negatively charged than in positively charged liposomes.
The fluorescence of both the monoamine 9-amino-6-chloro-2-methoxyacridine and the diamine atebrin is quenched more extensively than that of 9-aminoacridine. Although the percentage of fluorescence quenching of both atebrin and 9-aminoacridine is dependent on the outside pH, the relationship between the fluorescence quenching of the two probes under similar conditions is not pH-dependent.
It is concluded that calculation of ΔpH from the percentage of fluorescence quenching of fluorescent amines is not meaningful, that the osmotic volume of chloroplasts is not involved in the quenching process and, consequently, that the interaction between the acridines and energized membranes is more likely to occur at the level of the membrane proper. 相似文献
In this report we investigated whether the modulation of drug permeability in Chinese hamster ovary (CHO) cells was an energy-dependent process. We observed that (1) in the absence of glucose, metabolic inhibitors such as cyanide, azide, and dinitrophenol stimulated the uptake of [3H]colchicine and other drug; (2) cyanide-induced stimulation of drug uptake could be prevented by the presence of metabolizable sugars such as glucose and ribose; (3) cyanide-treated cells were fully viable; (4) on the addition of cyanide and glucose the kinetics of drug permeability changes were very rapid. These data are consistent with the hypothesis that an energy-dependent membrane barrier against the uptake of a variety of drugs was operative in CHO cells.The nature of this energy-dependent membrane barrier was examined in colchicine-resistant mutants (CHRC4 and CHRC5 cells) previously characterized as membrane mutants with greatly reduced drug permeability (Ling and Thompson, (1974) J. Cell Physiol. 83, 103–116). The mutants were more refractile to the cyanide-induced stimulation of drug permeability but more sensitive to the glucose prevention cyanide-induction. In the presence of cyadine, the uptake rate of [3H] colchicine by CHRC4 cells increased by about 100-fold and approached a rate similar to that of wild-type cells. These results suggest that the colchicine-resistant mutants may be altered in their energy-dependent modulation of drug permeability. 相似文献
Analogs of 4-androstene-3,17-dione and testosterone were tested as substrates for the aromatizing enzyme complex of human placenta. Compounds modified in rings B, C and D were found to be aromatized via a pathway similar to that postulated for 4-androstene-3,17-dione and testosterone, in which oxidation to the 19-hydroxy and 19-oxo (or corresponding gem-diol) intermediates occurs. No evidence of additional intermediates was obtained. 相似文献
Non-photochemical quenching (NPQ) of chlorophyll fluorescence is the process by which excess light energy is harmlessly dissipated within the photosynthetic membrane. The fastest component of NPQ, known as energy-dependent quenching (qE), occurs within minutes, but the site and mechanism of qE remain of great debate. Here, the chlorophyll fluorescence of Arabidopsis thaliana wild type (WT) plants was compared to mutants lacking all minor antenna complexes (NoM). Upon illumination, NoM exhibits altered chlorophyll fluorescence quenching induction (i.e. from the dark-adapted state) characterised by three different stages: (i) a fast quenching component, (ii) transient fluorescence recovery and (iii) a second quenching component. The initial fast quenching component originates in light harvesting complex II (LHCII) trimers and is dependent upon PsbS and the formation of a proton gradient across the thylakoid membrane (ΔpH). Transient fluorescence recovery is likely to occur in both WT and NoM plants, but it cannot be overcome in NoM due to impaired ΔpH formation and a reduced zeaxanthin synthesis rate. Moreover, an enhanced fluorescence emission peak at ~679?nm in NoM plants indicates detachment of LHCII trimers from the bulk antenna system, which could also contribute to the transient fluorescence recovery. Finally, the second quenching component is triggered by both ΔpH and PsbS and enhanced by zeaxanthin synthesis. This study indicates that minor antenna complexes are not essential for qE, but reveals their importance in electron stransport, ΔpH formation and zeaxanthin synthesis. 相似文献