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1.
Base dynamics of nitroxide-labeled thymidine analogues incorporated into (dA-dT)n by DNA polymerase I from Escherichia coli 总被引:1,自引:0,他引:1
Nitroxide-labeled thymidine substrates (dL) for Escherichia coli DNA polymerase I (pol I) were used to synthesize spin-labeled alternating double-stranded copolymers with (dA-dT)n as a template. All dL substrates use an alkane or alkene tether substituted into the 5-position of the pyrimidine ring to link a five- or six-membered ring nitroxide to the pyrimidine base. The kinetics of dL incorporation show some tether dependence with respect to tether length and tether geometry. The electron spin resonance (ESR) spectra of (dA-dT,dL)n duplexes directly formed by polymerization with pol I are compared with the ESR spectra of (dA)n(dT,dL)n duplexes, which are obtained after annealing of nitroxide-labeled single strands with complementary unlabeled single strands. The ESR spectra indicate that nitroxide-labeled analogues with tethers short enough to let the nitroxide ring reside in the major groove are excellent reporter groups for monitoring hybridization. A small difference between the ESR line shapes of the alternating duplexes (dA-dT,dL)n and the homopolymer duplexes (dA)n(dT,dL)n containing the same dL is detectable, suggesting the presence of subtle differences in the base dynamics between both systems. Computer simulation of the ESR spectra of the (dA-dT,dL)n duplexes was successful with the same motional model reported earlier [Kao, S.-C., & Bobst, A.M. (1985) Biochemistry 24, 5465-5469]. The thymidine motion arising from tilting and torsion of base pairs and base twisting in (dA-dT)n is similar to that in (dA)n(dT)n and is of the order of 4 ns. 相似文献
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Evaluation of growth rate in adhering cell cultures using a simple colorimetric method 总被引:2,自引:0,他引:2
B Pelletier F Dhainaut A Pauly J P Zahnd 《Journal of biochemical and biophysical methods》1988,16(1):63-73
In this paper we present a simple colorimetric method for evaluating cell growth in adhering cell cultures. This technique is based on the staining of basophilic cellular compounds (mainly nucleic acids) with methylene blue. To check its reliability, we have compared the quantity of dye fixed by the cells with the number of cells released from the culture substratum by trypsinization, and with the total cellular protein content determined by the Lowry's method. In these experiments we have used human skin fibroblasts and human epithelial cells derived from the epithelium of the full-term umbilical cord. Like other alternative methods, the methylene blue colorimetric technique can be used in 96-well microplates and allows the rapid collection of large amounts of data. As an illustration, we report on the selection of optimal composition of culture medium for human skin fibroblasts. 相似文献
4.
Zusammenfassung Es wurde die Röntgenstrahleninaktivierung von Laktatdehydrogenase in Rattenleber und Invertase in Hefe untersucht. Die Inaktivierung war in den feuchten lebenden Zellen und in den getrockneten Zellen praktisch gleich, obwohl ein Beitrag der indirekten Strahlenwirkung in den feuchten Zellen zu erwarten war.Auszugsweise vorgetragen auf dem XI. Internationalen Kongreß für Radiologie, Rom, September 1965 相似文献
5.
Summary The natural radionuclide content of mineral phosphate fertilizers has been determined gammaspectrometrically. The investigations comprised ca. 70% of the mineral phosphate fertilizers authorized and used in the Federal Republic of Germany (FRG). At maximum, we found specific activities of 62 nCi Unat/kg, 23 nCi226Ra/kg, 1.6 nCi Thnat/kg and 262 nCi40K/kg. The mean values, weighted by the percentual agricultural consumption of the main phosphate fertilizer groups in 1973/74 and related to the phosphate content, amounted to 58, 40, 2, and 584 nCi/kg P2O5 for Unat,226Ra, Thnat, and40K respectively. This resulted in an annual distribution due to phosphate fertilizing of about 3.9 µCi Unat, 2.7 µCi226Ra, 0.1 µCi Thnat, and 39.9 µCi40K per ha of arable or pasture land in 1973/74 on the average. From these values the air dose rates over agricultural areas have been estimated under extreme conservative assumptions resulting in an additional external exposure of members of the population of 0.02 mrd/a on the average and 0.4 mrd/a in the region of highest phosphate fertilizing intensity. If it is assumed that radium contained in phosphate fertilizers were completely accumulated in the soils during the last 80 years, this value would be raised to 0.3 mrd/a on the average. The occupational external radiation exposure due to natural radionuclides contained in phosphate fertilizers was estimated to be 0.1 mrd/a on the average and 2.3 mrd/a at maximum for persons working in agriculture. These estimates show that natural radionuclides in phosphate fertilizers contribute but very little to the mean terrestrial radiation exposure of the population which is 50 to 55 mrd/a in Germany. Only for the small group of persons working in fertilizer production plants or storehouses a significant increase of the external radiation exposure has to be expected which could reach a doubling of the mean natural exposure value.Dedicated to Prof. Dr. H. Muth on the occasion on his 60th birthday 相似文献
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Albert M. Bobst Shih-Chung Kao Elisabeth V. Bobst Gary T. Pauly 《Journal of biomolecular structure & dynamics》2013,31(2):249-260
Abstract Stoichiometric amounts of poly-L-lysine were added to site-specifically spin labeled single stranded nucleic acids and the resulting complexes analyzed by electron spin resonance spectroscopy (ESR). The nucleic acids were spin labeled to different extents and with labels of varying tether length. The ESR data are used to determine nucleoside dynamics and some structural features in these complexes. It is concluded that two distinct base mobilities exist in the complexes; one set is characterized by a mean correlation time τR = 2 ns, and the other one by a τR ≤ 50 ns. A model is proposed which suggests that a poly-L-lys single stranded nucleic acid complex consists of low mobility segments flanked by more mobile bases. An interesting feature of the proposed model is its applicability to explain ESR data of single strand binding protein-spin labeled nucleic acid complexes, which can also be interpreted in terms of two distinct nucleoside mobility states. It is hypothesized that this phenomenon could be of biological significance for the release of protein ligands from a protein-nucleic acid complex. 相似文献
8.
M. Ardhaoui A. Falcimaigne J.M. Engasser P. Moussou G. Pauly 《Biocatalysis and Biotransformation》2013,31(4):253-259
Enzymatic acylation of rutin and esculin with aromatic, aliphatic and aryl-aliphatic acids using Candida antarctica lipase in tert amyl alcohol as solvent was investigated under low water content. Whatever the acyl donor used, the conversion yields and initial rates for esculin were higher than for rutin. For a given flavonoid, the performance of these reactions depended on the acyl donor structures. For aliphatic acids, conversion yields and initial rates of both flavonoids were respectively in the ranges of 68–90% and of 9.5×10?2–72×10?2 mmol l?1 h?1. For aromatic acids, the reaction occurred only with the aryl subgroup (cinnamic, hydrocinnamic, 3,4-dihydroxyhydrocinnamic and 4-hydroxyphenyl acetic acids) and was drastically influenced by the presence of side chain and substitution patterns of the aromatic ring. Except for hydrocinnamic acid (75%, 23.4×10?2 mmol l?1 h?1), with these acids the conversion yields and initial rates were lower and in the range of 10–45% and of 0.7×10?2 to 12.1×10?2 mmol l?1 h?1. Unsaturation of the side chain of the hydrocinnamic acid decreased the esculin conversion rate from 75 to 13% and initial rate from 23.4 to 1.76×10?2 mmol l?1 h?1. The presence of hydroxyl or nitro-groups on the aromatic ring of the aryl aliphatic acid also reduced conversion yields and initial rates. Even without a spacer, the non-phenolic ring acid (quinic acid) was reactive and lead to conversion yields of about 20 and 23% respectively for rutin and esculin. 相似文献
9.
Fisheries catches from Pacific Island coral reefs are rarely recorded in official statistics. Reconstruction of catch estimates with limited hard data requires interpolation and assumptions, justifiable only by the unsatisfactory alternative of continued substitution of zero catches, a common policy interpretation for ‘no data’. Uncertainties associated with reconstructions are high, requiring conservative estimation. American Samoan domestic fisheries consist of an artisanal, small-boat sector, whose commercial catches are reported, and a shore-based subsistence sector, with no regular reporting. Our catch reconstruction (with large pelagic species removed) suggested a 79% decrease in catches between 1950 (752 t) and 2002 (155 t). Accounting for rapid human population growth on the main island, the per capita catch rate may have declined from 36.3 kg·person−1 year−1 in 1950 to 1.3 kg·person−1 year−1 by 2002, while the catch rate for the inhabited outer islands has been independently reported as 58.6 kg·person−1 year−1. Catch per area of coral reef (to 50-m depth) may have declined from 5.5 to 0.7 t km−2 year−1 for the main island, and from 9.1 to 4.9 t km−2 year−1 for the outer islands, for 1950 and 2002, respectively. Summed for 1950–2002, our reconstruction suggested a 17-fold difference between reconstructed estimates and reported statistics. 相似文献
10.
Woo Hwi Yang Oliver Heine Sebastian Pauly Pilsang Kim Wilhelm Bloch Joachim Mester Marijke Grau 《PloS one》2015,10(4)