FAR UV IRRADIATION OF DNA IN THE PRESENCE OF PROTEINS, AMINO ACIDS OR PEPTIDES

Abstract— The DNA of bacteriophage SP02c12 was subjected to 254 nm irradiation in solutions containing lysozyme or histone. In these solutions, the protein-DNA mass ratios and the ionic strengths of the solvents were varied to change the amount of protein associated with the DNA. Lysozyme-DNA binding constants were measured under the same conditions. The sensitivity of phage DNA to biological inactivation by UV increased as the amount of lysozyme bound per DNA strand increased. Although binding constants could not be measured for the DNA-histone interaction, this protein had a protective effect which was greater under conditions which cause enhanced binding. No crosslinking of either protein could be detected even at doses ten-fold greater than those giving a surviving fraction of 0.01.
Irradiation was also performed in the presence of various amino acids and short peptides. These were chosen to include amino acids which: (1) are positively charged, (2) absorb UV of this wavelength or (3) form UV-induced crosslinks to DNA. None of the amino acids tested affected sensitivity of the DNA to biological inactivation. Peptides containing a UV-absorbing amino acid and a positively charged amino acid enhanced sensitivity. For each of these peptides, a mixture of the constituent amino acids had the same effect as the peptide itself. Under the conditions used, no evidence for formation of DNA-amino acid crosslinks was found. The results indicate that proteins and peptides can sensitize DNA to UV inactivation by mechanisms other than covalent crosslink formation. Such mechanisms could include energy or electron transfer or alterations in the conformation of the DNA.     

ACTIVATION OF THE HUMAN IMMUNODEFICIENCY VIRUS PROMOTER BY UVA RADIATION IN COMBINATION WITH PSORALENS OR ANGELICINS

Abstract— The effects of mono- and bifunctional furocoumarins plus UVA radiation (PUVA and related treatments) on the human immunodeficiency virus-1 (HIV-1) promoter were studied using HeLa cells stably transfected with the chloramphenicol acetyl transferase gene under the control of the HIV-1 promoter. The experiments were performed with three psoralens (5-methoxypsoralen, 5-MOP; 8-methoxypsoralen, 8-MOP; and 4′-aminomethyl-4,8,5′-trimethyl-psoralen, AMT) and four angelicins (angelicin; 4,5′-diniethylangclicin, 4,5′-DMA; 6,4′-dimethylangelicin, 6,4′-DMA; and 4,6,4′-trimethylangelicin, TMA). The drugs alone and UVA radiation alone showed no erect on the HIV promoter. However, when the cells were incubated with the furocoumarins at 0.1–40 μg/mL and then irradiated. the HIV promoter was activated in distinct fluence ranges, i.e. (1) no promoter activity was discernible at low fluences (e.g. at 0.1 μg/mL of 8-MOP up to 100 kJ/m²), (2) as the fluence was increased, the promoter activity increased to reach a maximum (10–50-fold with respect to the unexposcd samples), and (3) as the fluence was further increased, the promoter activity decreased. Similar (although shifted on the fluence scale) pattcrns were observed with either > 340-nm UVA radiation or with UVA radiation contaminated with a small amount of UVB radiation (typical for PUVA lamps). The effective fluences were inversely related to the drug concentration. Experiments with 5-MOP and 8-MOP indicated reciprocity of the drug concentration and radiation hence. The HIV promoter response patterns were similar for monofunctional angelicins and bifunctional psoralens. This indicated that the furocoumarin-DNA crosslinks are not a prerequisite for the promoter activation and that the monoadducts suffice to elicit the HIV promoter response. The HIV promoter-activating effectiveness of diKcrent drugs correlated with their photosensitizing potential. Thus, among psoralens the effectiveness order was AMT >. 5-MOP >8-MOP, and among angelicins: TMA > 6,4′-DMA > 4,5′-DMA > angelicin. The ektiveness did not vary substantially for 5-MOP, 8-MOP, 4,5′-DMA, and 6,4′-DMA. The combined drug and UVA radiation doses were higher than those that elicit cellular responses or those that may be received by the human white blood cells during cxtracorporeal PUVA therapy (photopheresis).     

EFFECT OF TEMPERATURE AND pH ON THE QUENCHING OF TRIPLET LUMIFLAVIN IN THE PRESENCE AND ABSENCE OF FERRI- AND FERROCYANIDE

Abstract— –Flash photolysis at 450 nm over the temperature range 0.8–60°C was used to determine Arrhenius parameters for the first and second order disappearance of triplet lumiflavin (1.66 µ .M ) at a flash energy of 2 kj in deaerated phosphate buffer at varying pH:
³Lf → Lf₀
³Lf +³Lf → Lf₀+ Lf₀
Arrhenius parameters were also determined for the pseudo first-order quenching of triplet lumiflavin by 10 µ M ferri- and ferrocyanide ions,
³Lf + Fe³⁺→ Fe³⁺→ Lf₀+ Fe³⁺ (energy transfer)
³Lf + Fe²⁺→ Lf^-+ Fe³⁺ (electron transfer)
and for disappearance of the semireduced lumiflavin in the presence of ferrocyanide at pH 6.8, by the second-order reaction
Lf^-+Lf ^-→ Lf₀+ Lf⁼.     

THE MODIFICATION OF RIBONUCLEASE-A BY NEARULTRAVIOLET IRRADIATION IN THE PRESENCE OF PSORALEN

Abstract— Ribonuclease A is inactivated when irradiated under oxygen by UV-A light in the presence of psoralen. The rate of inactivation is greatly reduced by sodium azide. ascorbate or nitrogen, whereas the substrate gives only very limited protection. A ribonuclease sample modified to 40% remaining activity presented a significant modification of amino acid residues known to be sensitive to oxidation and 1.4 mol of bound psoralen per mol of protein. The secondary structure of the enzyme, as assessed by circular dicroism was not changed by irradiation; neither was aggregation of the enzyme to a higher mol wt evident. Studies on the tryptic peptides fractionated by high performance liquid chromatography showed that the photomodification occurs with very low selectivity. All the five peptides containing hystidine, tyrosine and methionine residues were greatly modified, although two, those containing histidine residues 12 and 119 in the sequence, amino acids known to be involved in the catalytic activity of ribonuclease. are modified to a greater extent. The protein bound psoralen. revealed by radioactivity in the HPLC eluate, was not found associated to only one or few peptide peaks but spread on a large zone of elution.     

EXTENSIVE CROSSLINKING BETWEEN SUBUNITS OF OLIGOMERIC PROTEINS INDUCED BY FUROCOUMARINS PLUS UV-A IRRADIATION

Abstract— Exposure of oligomeric proteins to UV-A irradiation in the presence of various furocoumarins results in extensive crosslinking of the enzyme subunits. The mechanism by which these covalent bridges are formed does not involve the furocoumarin moiety as the crosslinking agent, but depends upon oxidative processes, mainly involving singlet oxygen and radical mediated processes not involving oxygen. The formation of crosslinks is not per se responsible for the inactivation of enzymes since in glutamate dehydrogenase the two reactions proceed at the same rate, in catalase it is not accompanied by enzyme inactivation and in alcohol dehydrogenase the inactivation rate is higher than polymerization.     

PROTECTIVE EFFECT OF SELENIUM AND ZINC ON UV-A DAMAGE IN HUMAN SKIN FIBROBLASTS

Ultraviolet A radiation participates in cytotoxicity and carcinogenesis of the skin by a mechanism involving the generation of reactive oxygen species. Endogenous antiradical defense systems utilize metalloenzymes including Se-dependent glutathione peroxidase and Cu and Zn superoxide dismutase. The aim of the present work was to determine the protective effect of two trace elements, Se and Zn, on cultured human diploid fibroblasts exposed to UV-A radiation (broad-spectrum source with a maximum intensity at 375 nm). Selenium in the culture medium (0.1 mg/L) in the form of sodium selenite increased the synthesis and activity of glutathione peroxidase by 60.5% in the absence of exposure to UV-A radiation and by 35% after irradiation with 5 J/cm² ( P = 0.043). The presence of this element significantly increased the survival of UV-A-irradiated fibroblasts ( P < 0.0001). This confirms the essential role of Se in the detoxifying activity of the enzyme. In addition, thiobarbituric acid-reacting substances (TBAR), which are lipid peroxidation markers, decreased in the presence of exogenous Se:—19% and -22% without irradiation and after irradiation with 5 J/cm² ( P = 0.056). When Zn was added at the dose of 6.5 mg/L as ZnCl₂, fibroblasts subjected to oxidizing stress induced by UV-A were protected from cytotoxicity ( P <0.0001). The TBAR production decreased significantly: -33% without irradiation and -34% after irradiation with 5 J/cm² ( P = 0.008). Superoxide dismutase activity, however, decreased after supplementing with Zn: - 26% without irradiation and - 20% after UV-A irradiation ( P = 0.017). The antioxidant properties of Zn are thus apparently independent of superoxide dismutase activity.     

辐照亚硫酸纸浆结晶度的研究

<正> 纤维素分子中含有大量的羟基,易形成氢键,是结晶性很高的聚合物,无论在研究工作和工业生产中,这一性质都受到高度重视。纤维素在辐照场中的结晶行为,也是被研究的重点。从已发表的研究结果看,辐照纤维素的结晶行为有很大的差别,据有关文章报道,在γ射线和电子束射线的辐照场中,纤维素的结晶结构没有变化。例如,美国     

在DBU存在下酞菁锰配合物的合成及溶剂影响

在强有机碱１，８－二氮杂双环［５，４，０］十一－７烯（简称ＤＢＵ）存在下，ＭｎＣｌ２与邻二氰基苯在某些二元醇溶剂中可高效合成Ｍｎ（Ⅱ）Ｐｃ（Ｐｃ表示酞菁），同时研究了不同溶剂对合成产物的物种及产率的影响和不同氧化态锰的酞菁配合物在一些醇溶剂中的互变现象     

STUDIES ON LYSOZYME: EFFECT OF VISIBLE LIGHT ON THE CONFORMATION OF THE ENZYME IN THE PRESENCE OF METHYLENE BLUE

Abstract— Lysozyme undergoes conformational changes when exposed to visible light in the presence of methylene blue. Ultracentrifugation, gel filtration and end group analysis showed that no peptide bond was cleaved. About 30 per cent of the tryptophan residues were, however, modified. The enzymic activity decreased by 50 per cent. The sedimentation coefficient and helical content decreased; the extinction in the u.v. region between 185 to 210 nm increased. The photooxidized enzyme was more susceptible to trypsinolysis than the native enzyme.     

THE EFFECT OF IRRADIATION ON THE POLYETHERKETONE WITH CARDO GROUP

The effects of irradiation on the polyetherketone with cardo group (PEK-C) were studied. It was found that PEK-C can be crosslinked by irradiation under vacuum, while degradation reaction occurred in PEK-C at room temperature in the presence of air. Moreover. it was also found that Ts value of the crosslinked PEK-C at high temperature is higher than that at room temperature in the case of the same gel content, whose value is about 8℃higher than that of unirradiated PEK-C. The gelation dose of PEK-C at 300℃under vacuum is 1.5×10~4 Gy, which is about hundred times smaller than that at room temperature.     

PENETRATION OF SOLAR UV IRRADIATION IN COASTAL LAGOONS OF THE SOUTHERN BALTIC SEA AND ITS EFFECT ON PHYTOPLANKTON COMMUNITIES

Abstract Direct measurements of solar UV irradiance at the surface and in the water column were made in several coastal lagoons near the islands of Hiddensee and Rügen in spring and summer 1993 using an internally frosted quartz sphere at the end of a 10 m quartz fiber cable connected to a double monochromator spectroradiometer. The measurements showed a nearly uniform vertical distribution of the absorption coefficient in the whole water column due to effective mixing, shallow depths and little temperature and salinity density stratifications. There was an increase in the absorption coefficient at 310 nm from 5 m⁻¹ in the outer lagoons to 12 m⁻¹ in the open inner lagoons and up to 29 m⁻¹ in the inner lagoons without water exchange with the Baltic Sea as a result of different amounts of UV-absorbing substances such as yellow substance (Gelbstoff) and seston. Depending on the specific concentration of the absorbents, the solar UV-B irradiance decreases to 1% after penetration into the water column between 0.2 m (inner lagoons without water exchange), 0.4 m (inner lagoons with water exchange), and 1 m (outer lagoons), whereas UV-A penetrates on the order of several meters. As phytoplankton populates the top layers of the water column it may be affected by increased solar UV irradiation even in these turbid coastal waters.     

THE EFFECTS OF MAGNETIC FIELD ON GRAFT COPOLYMERIZATION OF METHYL METHACRYLATE ONTO POLYVINYL ALCOHOL IN THE PRESENCE OF BENZOPHENONE DURING UV IRRADIATION

The effects of magnetic field on the graft ratio and stereoregularity of grafts of PVA-g-MMA in the presence ofbenzophenone during UV irradiation are discussed. By means of IR, it was found that the graft ratio was increased with the increment of magnetic field strength. Furthermore, application of relative weak magnetic field of 0.4 Tesla had been shown to substantially enhance the stereo-regularity of graft copolymer. The maximum stereo-regularity appeared when the graft ratio approached to 85% with the magnetic field of 1.2 Tesla (T). The resistance to moisture and heat resistance of the grafted copolymer in the presence of magnetic field were also improved.     

γ-射线辐照对尼龙6纤维结构的影响

<正> 过去的大多数研究表明,高能射线辐照会引起聚乙烯及聚丙烯等稀烃类聚合物分子链交联及降解,而且会导致结晶度下降。张利华等人最近的研究表明,对于尼龙1010塑料,仅当辐照剂量高到一定程度后,辐照才会破坏结晶。此外,Stowe等人曾经研究过近紫外线对尼龙66纤维结构的影响,结果表明,在空气气氛中的近紫外辐照主要导致尼龙66纤维降解,并使纤维结晶度提高。有关高能射线辐照对高度取向的合成纤维结构及性能的影响的研究尚不多见。本工作将讨论γ-射线对于尼龙6纤维结构的影响。     

FACTORS INFLUENCING THE NEAR-UV EFFECT ON DNA IN THE PRESENCE OF 8-METHOXYPSORALEN: RENATURABILITY OF DNA.

Factors affecting the near-UV induced reaction of calf thymus DNA in the presence of 8-methoxypsoralen were examined. DNA became more renaturable with increasing concentration of 8-methoxypsoralen and fluence. Existence of paramagnetic ions, Mn²⁺, Co²⁺, or Ni²⁺, largely repressed the photoreaction. It was ascertained that the highest renaturability was attained by the illumination with light between 310 and 345 nm.     

THE PHOTODYNAMIC EFFECT OF ROSE BENGAL ON PROTEINS OF THE MITOCHONDRIAL INNER MEMBRANE

Photodynamic action promoted by Rose Bengal was evaluated in solutions of unsaturated fatty acids or histidine, and on beef heart submitochondrial particles. Rose Bengal-promoted photooxidation of histidine was mainly due to the opening up of the imidazole ring by singlet oxygen. Photosensitization of polyunsaturated fatty acids (PUFA) resulted in oxygen consumption and thiobarbituric acid-reactive substances (TBARS) formation, the extent of which was linearly related to the increasing degree of unsaturation. Photosensitization of submitochondrial particles caused oxygen consumption and TBARS production. These processes involved two different reaction components: during the first, most of the mitochondrial proteins were inactivated, the most sensitive being succinate dehydrogenase and cytochrome c. The values for the rate ratios of [TBARS] formation/[O2] consumption for the first and second phase were 0.36 and 1.32%, respectively, pointing to a larger contribution of lipid peroxidation during the second phase. The calculation of the rate constants for reaction of singlet oxygen with mitochondrial proteins suggests that singlet oxygen is more reactive towards proteins than to PUFA. The biological role of this selectivity is discussed in terms of the mitochondria as one of the first targets for photosensitized reactions.     

在混合表面活性剂存在下二溴邻硝基苯基荧光酮吸光光度法测定钒的研究

在pH6.4的柠檬酸-Na_2HPO_4缓冲介质中,V(V)与二溴邻硝基苯基荧光酮和CPB反应生成红色多元配合物,建立了吸光光度测定微量钒的方法.配合物最大吸收波长为595nm.表观摩尔吸光系数为6.5×10~4.钒量在0～20μg/25ml范围内符合比耳定律.经测定配合物组成比为V(V):DBo-NPF:CPB=1:4:4.采用MIBK溶剂萃取使钒与干扰元素分离后.方法用于合金钢中微量钒的测定,回收率为99.2%～100.1%,相对标准偏差1.2%～4.2%.     

ON THE KINETICS AND MECHANISM OF THE COPOLYMERIZATION OF ACRYLONITRILE WITH STYRENE IN THE PRESENCE OF COPPER CHLORIDE

The copolymerization of styrene (St) and acrylonitrile (AN) complexed with CuCl_2 monomer by a free radicalmechanism was performed using benzoyl peroxide as an initiator at 65℃ under N_2 atmosphere for 150 min. The rate ofpolymerization (R_p) was found to increase linearly with the concentration (in mol/L) of CuCl_2, AN and St through scalingrelations. The activation energy of the copolymerization process in the presence and absence of CuCl_2 was found to be46.5 kJ/mol and 102 kJ/mol, respectively. The viscosity average molecular weigh of the copolymer and the k_p~2/k_t ratio weredctermired to further assess the accelerating effect of CuCl_2 on the copolymerization process. The copolymerization processin the presence of CuCl_2 has a radical complex mechanism.     

EFFECTS OF HEMATOPORPHYRIN DERIVATIVE ON THE CELLULAR ENERGY METABOLISM IN THE ABSENCE AND PRESENCE OF LIGHT

Effects of Photofrin II on energy metabolism and metabolic viability were studied in a mammalian transformed cell line (BHK-21) in dark and after photo-irradiation with visible light. Cells were allowed to accumulate Photofrin by incubating for 4 h in buffer containing Photofrin (5-60 micrograms/ml). The results show that Photofrin significantly affects the cellular energy metabolism even in the absence of light; activity of cytochrome c oxidase is decreased and glucose utilization and lactate production (glycolysis) are increased. Irradiation with light resulted in a significant decrease in the activity of cytochrome c oxidase, glycolysis, ATP content, energy charge, ratios of adenine nucleotides like ATP/ADP, ATP/AMP and cell viability (dye exclusion test). Presence of inhibitors of energy metabolism, potassium cyanide (respiration) and 2-deoxyglucose (glycolysis), further enhanced the cytotoxic effects induced by hematoporphyrin derivative and light.     

以氯代钒酸酯-三异丁基铝催化体系合成丁二烯-丙烯交替共聚物的研究

本文比较了各种氯代钒酸酯-三异丁基铝体系催化丁二烯、丙烯交替共聚的活性,得出氯代钒酸二新戊酯-三异丁基铝体系催化活性最高。能制得高分子量的交替共聚物。研究了该催化体系的特点和聚合规律及提高共聚物[η]的途径。在-76--45℃温度范围内可以制得正[η]为1.7-2.6dl/g的丁丙交替共聚物,单体转化率在80％以上。通过分析鉴定,证明产物交替度在95％左右,反式1,4丁二烯组分含量在95％以上。     

EFFECT OF BASE SEQUENCE ON THE ULTRAVIOLET IRRADIATION PRODUCTS OF DOUBLE-STRANDED POLYNUCLEOTIDES CONTAINING BROMOURACIL AND ADENINE

Abstract —The effects of ultraviolet irradiation of double-stranded synthetic polynucleotides containing BrU and A have been investigated. Homopolymer pairs and alternating copolymers composed of either ribo- or deoxyribo-nucleotides were prepared and were irradiated with either 313 nm or ˜ 285 nm light. Strand separation and a modest amount of strand breakage followed irradiation of the homopolymer pairs. Changes in the ultraviolet absorption spectra of the polymers during irradiation reflected the sum of hyperchromic increases caused by progressive strand separation and loss of absorbance caused by photoproduct formation. Extensive debromination occurred. An RNase digest of irradiated poly(rA)–poly¹⁴C(rBrU), analysed by column chromatography, showed components similar to those found previously upon irradiation of single-stranded poly(rBrU). Little photoproduct was released by RNase digestion as mononucleotides. The major photoproduct was in the dinucleotide fraction, and may be 5,5'-diuracil. Base sequence had a profound effect on the sensitivity of the polynucleotides. Irradiation of alternating copolymers with doses of light comparable to those that produced major photochemical changes in the homopolymer pairs brought about little if any change in the copolymers of alternating base sequence.