Temperature-dependence of UV laser one-electron oxidative guanine modifications as a probe of local stacking fluctuations and conformational transitions

By monitoring R(pip)/R(Fpg), i.e. the relative sensitivity to hot piperidine and to formamidopyrimidine DNA glycosylase (Fpg protein) of the guanine lesions induced in DNA exposed to UV laser irradiation, we have previously observed that the formation of the two major types of one-electron oxidative guanine modifications, oxazolone and 7,8-dihydro-8-oxoguanine (8-oxodG), depends on DNA conformational features. While oxazolone is largely predominant at each site of single-stranded DNA (R(pip)>R(Fpg)), 8-oxodG is the major lesion at most of the sites of double-stranded DNA (R(pip)R(Fpg) at 20 degrees C and the ratio R(pip)/R(Fpg) does not vary significantly during the melting process. Interestingly, these guanine residues display a high sensitivity to dimethyl sulfoxide methylation while the opposite cytosine residues are unsensitive, suggesting that the prevalence of R(pip) over R(Fpg) is related not to base-pairing disruption but rather to the local helical alteration of the B-DNA stacking geometry. This leads us to propose that the slight variations in the ratios R(pip)/R(Fpg) observed, at individual sites, at temperatures below the helix-coil transition reflect local small-scale breathing motions, unstacking single dinucleotide steps prior to opening. Our results thus support the view that the temperature dependence of the ratio of R(pip)/R(Fpg) at sites of B-DNA provides a sensitive probe of the DNA internal local thermal stability and are discussed in relation with the mechanisms proposed for the intramolecular rearrangement of the guanyl radical.     

An analysis of the action of light on betalain synthesis in the seedling of Amaranthus caudatus,var. viridis

Accumulation of betalain (amaranthin) in the seedling of Amaranthus caudatus, var. viridis, is inducible by light. Since the apparent lag-phase of amaranthin accumulation after the onset of light is of the order of 3 h, light induction experiments could be performed up to 3 h after the onset of light without interference with actual synthesis. The intricate induction phenomena can be explained as follows: The inductive light operates through phytochrome and through a blue/UV photoreceptor (cryptochrome). A phytochrome-dependent High Irradiance Reaction is of minor importance. However, there is a strong, specific interaction between the light effects mediated through phytochrome and cryptochrome in the sense that the extent of the reversible response — (response obtained with a particular light treatment terminated with a saturating red light pulse) minus (response obtained with the same light treatment when terminated with a saturating 756 nm light pulse) —increases with increasing P_fr level and total fluence rate during the induction period. It is concluded that light induced amaranthin synthesis is, in fact, a convenient biochemical model system of photomorphogenesis in the case when phytochrome and cryptochrome operate simultaneously in mediating photomorphogenesis. Abbreviations: see Table 1 and 2 Materials and Medthods     

Lignin-carbohydrate complexes from Lolium perenne

Lignin-carbohydrate and lignin-hemicellulose complexes were extracted sequentially from four varieties of Lolium perenne which had previously been     

溶藻菌株NP23的紫外诱变选育

应用紫外诱变技术对溶藻菌株NP23进行紫外诱变处理。经过粗筛后,从8株诱变株中选出2株对绿藻中小球藻和蓝藻中惠氏微囊藻的去除效果明显优于原始菌株的突变株NP23-1和NP23-4,其溶藻率(叶绿素a的去除率)比原始菌株提高30%-35%。连续6代测试,2株诱变菌株NP23-1和NP23-4溶藻率都很稳定,表明所得突变株是比原始菌株更优秀的溶藻菌株。     

Characterization of human serum albumin's interactions with safranal and crocin using multi-spectroscopic and molecular docking techniques

Interaction mechanisms of human serum albumin (HSA) with safranal and crocin were studied using UV–Vis absorption, fluorescence quenching and circular dichroism (CD) spectroscopies as well as molecular docking techniques. Changes in absorbance and fluorescence of HSA upon interactions with both compounds were attributed to their binding to amino acid chromophores located in subdomains IIA and IIIA. Fluorescence secondary inner filter effect was excluded using 278 nm and 340 nm as the wavelengths of HSA's excitation and fluorescence while safranal and crocin absorbed at 320 nm and 445 nm, respectively. Stern-Volmer model revealed a static quenching mechanism involve the formation of non-fluorescent ground state complexes. Stern-Volmer, Hill, Benesi-Hilbrand and Scatchard models gave apparent binding constants ranged in 4.25 × 10³ - 2.15 × 10⁵ for safranal and 7.67 × 10³ - 4.23 × 10⁵ L mol^?1 for crocin. CD measurements indicated that 13 folds of safranal and crocin unfolded the α-helix structure of HSA by 7.47–21.20%. In-silico molecular docking revealed selective exothermic binding of safranal on eight binding sites with binding energies ranged in ?3.969 to ?6.6.913 kcal/mol. Crocin exothermally bound to a new large pocket located on subdomain IIA (sudlow 1) with binding energy of ?12.922 kcal/mol.These results confirmed the formation of HSA stable complexes with safranal and crocin and contributed to our understanding for their binding characteristics (affinities, sites, modes, forces … etc.) and structural changes upon interactions. They also proved that HSA can solubilize and transport both compounds in blood to target tissues. The results are of high importance in determining the pharmacological properties of the two phytochemical compounds and for their future developments as anticancer, antispasmodic, antidepressant or aphrodisiac therapeutic agents.     

Comparison of spontaneous and ultraviolet-induced mutagenesis on naked SV40 DNA and SV40 virus

Molecular aspects of mutagenesis in mammalian cells have been essentially analyzed using biological probes such as viruses and shuttle vector. Although the main data concerning the specificity of carcinogen-induced mutations are similar, the observed spontaneous mutation frequencies are significantly different when using one or the other model. This frequency is considerably higher with shuttle vectors than with viruses. We have performed an analysis of mutagenesis in order to determine if the obligatory transfection step associated with shuttle vector technology was responsible for the high mutation frequency found with these molecules. For this purpose simian virus 40 (SV40) genome used as virus or as naked DNA was introduced into permissive cells by viral infection or DNA transfection respectively. Our results show that transfection alone does not induce a higher mutation frequency on SV40 DNA the virus infection. Moreover, we have shown that the ultraviolet-light induced mutation spectrum was similar on the SV40 VP1 gene after viral infection or DNA transfection.     

激光预处理可保护蚕豆细胞免受UV—B辐射的损伤

当蚕豆的胚被 He- Ne激光 (632 .8nm,1 .63J· mm- 2 )照射 5min或被 CO2 激光 (1 0 60nm,2 .53J· mm- 2 )照射 1 min后 ,将其置入 Knop营养液中进行恒温培养。当蚕豆的上胚轴长到大约 3cm时 ,在光背景 (PAR)为 70μmol· m- 2 · s- 1条件下 ,分别用 1 .0 2、3.0 3、4.52 k J· m- 2 的 UV- B辐射蚕豆的上胚轴 7h。根据蚕豆丙二醛 (MDA)、抗坏血酸 (As A)和 UV- B吸收物的含量变化 ,来测试激光对 UV- B照射蚕豆的上胚轴的保护作用。结果显示 :激光预处理可保护蚕豆上胚轴对 UV- B辐射的作用。与对照组 (没有用 UV- B或激光照射 )、UV- B单独照射组比较 ,在激光预处理的条件下 ,MDA的含量明显减少 ,As A和 UV- B吸收化合物的含量增加。如先用激光处理 ,然后再用 UV- B辐射 ,UV- B吸收物的含量将比单独用激光和 UV-B处理获得更好的改善。从而认为 ,激光预处理能增强植物对 UV- B的抵抗力。     

Biochemical and spectroscopic analysis of UV effects in the marine flagellate Cryptomonas maculata

The effects of solar and artifical ultraviolet radiation on the marine cryptoflagellate, Cryptomonas maculata, were studied. Even after short exposure to UV the accessory photosynthetic pigment phycoerythrin is bleached; likewise the fluorescence undergoes significant changes both in amplitude and in the maximal peak wavelength. In parallel, the photosynthetic oxygen production decreases rapidly during exposure. Gel electrophoresis and FPLC of membrane proteins show a significant decrease in chromoproteins after 2 h UV, which is confirmed by fluorescence excitation and emission spectra of the FPLC fractions.Abbreviations APS ammonium persulfate - DCMU 3-(3,4dichlorophenyl)1,1-dimethylurea; Emulphogen, polyoxyethylene 10 tridecyl ether - FPLC fast protein liquid chromatography - PMSF phenylmethylsulfonyl fluoride - SDS sodium dodecylsulfate - SDS PAGE sodium dodecylsulfate polyacrylamide gel electrophoresis - TEMED NN NNtetramethylethylene diamine - UV-A wavelength range between 320 nm and 400 nm - UV-B wavelength range between 280 nm and 320 nm Dedicated to the 60th birthday of Professor Dr. W. Wehrmeyer     

Thiol-stimulated secretion of riboflavin and porphyrins by Escherichia coli

Abstract Streptococcus lactis strain IL594 contains 9 plasmids, designated pIL1 to pIL9. On the basis of protoplast-induced curing experiments we showed that derivatives containing pIL7 (31 kb) were resistant to UV-irradiation while derivatives lacking pIL7 were sensitive. The pIL7-determined UV-protection was confirmed by cotransfer of the plasmid and of the character into a plasmid-free derivative of S. lactis IL594. Moreover, prophage induction required higher UV-fluence in this derivative carrying pIL7 than in the plasmid-free strain. This is the first report of a plasmid-mediated UV-protection in group N streptococci.     

Protoplast induction in Chlorella pyrenoidosa

Chlorella pyrenoidosa (UTEX 1230) cells in late log phase of growth were induced to form viable protoplasts by enzymatic digestion only when incubated in 2-deoxy-d-glucose (2DG) for 24 h. The combination of hemicellulase (4% w/v), Cellulysin (4% w/v), and glucuronidase (5% v/v) with 0.8 M mannitol and 8 mM CaCl₂ in modified Bristol's solution, was most effective for obtaining viable protoplasts as determined by light and electron microscopy, and vital staining with primuline (0.01% w/v). Resistance of cell walls to extensive extraction (acetolysis), and infrared analysis indicated that sporopollenin is a component of the cell wall. Transmission electron miscroscopy of acetolysed cell walls also allowed visualization of the laminate nature of the wall. This is the first report of successful induction of protoplasts from algae which contain sporopollenin in their cell walls.