ON THE MECHANISM OF RADIOSENSITIZATION BY 5-BROMOURACIL. THE OCCURRENCE OF DNA STRAND BREAKS IN U.V.-IRRADIATED PHAGE T4 AS INFLUENCED BY CYSTEAMINE

Abstract— U.V.-irradiation of phage T 4Bo^r results in a decrease in sedimentation rate of BU-DNA which is attributed to single- and double-strand breaks. No breaks could be observed in unsubstituted DNA. Cysteamine present during u.v.-irradiation is able to prevent double-strand breaks but does not influence the production of single-strand breaks measured by alkaline sucrose gradient centrifugation. The biological importance and nature of DNA strand breaks due to BU-incorporation as well as the action of the protective agent on these breaks and on the biological activity are discussed.     

COMPARISON OF PHAGE T4 denV ENDONUCLEASE V AND M. Luteus UV-DNA ENDONUCLEASE BY SEROLOGY AND DNA HYBRIDIZATION

Abstract Antibodies were raised in rabbits against purified endonuclease V, the product of the bacteriophage T4 denV gene, which incises DNA at the site of UV-induced pyrimidine dimers. These antibodies cross-reacted with the purified UV-DNA endonuclease from M. luteus in both enzyme-linked immunosorbent assay and Western assays. However, no sequence similarity was detected between the denV gene and M. luteus DNA by hybridization. The two endonucleaes are remarkably similar in enzymatic activity, and their antigenic similarities have been preserved despite differences in their DNA sequences.     

A KINETIC ISOTOPE EFFECT IN THE PHOTOREACTIVATION OF U.V.-INACTIVATED TOBACCO MOSAIC VIRUS RNA*

Abstract— Inactivation of tobacco mosaic virus RNA (TMV-RNA) by u.v. radiation is slower in D₂O than in H₂O, and TMV-RNA which has been inactivated in D₂O is photoreactivated faster (on Pinto bean) than TMV-RNA which has been inactivated in H₂O. The maximum amount of photoreactivation is unaffected by the solvent, H₂O or D₂O, present during irradiation. These deuterium isotope effects for inactivation and photoreactivation suggest that pyrimidine hydrates are photoreactivable lesions on Pinto bean.     

INHIBITION OF UV AND PSORALEN-PLUS-LIGHT MUTAGENESIS IN PHAGE T4 BY GENE 43 ANTIMUTATOR POLYMERASE ALLELES

Abstract— Mutagenesis by UV light or psoralen-plus-light was measured by increases in the reversion of nonsense mutants of phage T4. In the presence of either of two gene 43 antimutator polymerase alleles, ts CB120 or ts CB87, UV-induced reversion was inhibited. Likewise psoralen-plus-light mutagenesis was inhibited when the ts CB120 allele was present. These results imply that the gene 43 DNA polymerase has a role in the formation of mutations from the DNA lesions induced by UV and psoralen-plus-light.     

DIFFUSE SPECTRAL REFLECTANCE MODIFICATION OF SKIN BY U.V. IRRADIATION

Abstract— Irradiation of guinea pig skin in vivo with light shorter than 300 nm produced a significant decrease in 330–400 nm reflectance. A similar decrease in reflectance was seen by irradiation of excised guinea pig and human skin. Chemical or physical vasodilation had no effect on the 240–400 nm reflectance. Irradiation of guinea pig skin in vivo with light longer than 300 nm produced no change in 330–400 nm reflectance unless photosensitized with 8-MOP. U.V. irradiation of skin (in the presence of suitable endogenous or exogenous photo-sensitizers) may result in the formation of new products that effectively absorb 330–400 nm photons.     

BASE-SPECIFIC DAMAGE INDUCED BY 4-THIOURIDINE PHOTOSENSITIZATION WITH 334-nm RADIATION IN M13 PHAGE DNA

Abstract— Site-specific DNA damage caused by 334-nm radiation in the presence of the rare Escherichia coli base 4-thiouridine was investigated in vitro by detecting the sites of the termination of DNA synthesis with irradiated M13 phage DNA used as a template. Single-strand breakage was also examined. The results indicate that 334-nm radiation at very low fluences in the presence of 4-thiouridine induces termination of strand synthesis at thymine base sites and at the base immediately prior to thymine. Termination at these sites was diminished by treatment with hot piperidine. Strand cleavage by piperidine treatments was observed preferentially at the guanine site, but only after irradiation at much larger fluences. It is hypothesized that at low fluences 4-thiouridine forms photoadducts with thymine that block DNA synthesis, while at high fluences the guanine site is damaged via oxygen species.     

SYNERGISTIC LETHALITY OF PHAGE T7 BY NEAR-UV RADIATION AND HYDROGEN PEROXIDE: AN ACTION SPECTRUM

Abstract— Ozonation of valerophenone oxime o-methyl ether (4) produced a stereoisomeric mixture of crystalline dimeric valerophenone peroxides, 5a and 5b . N-n-butyl-N-methoxybenzamide (6) and N-methoxy-N-phenylvaleramide (7) along with valerophenone (1). Thermolysis of the higher melting peroxide 5a at 170–180°C, where a chemiluminescence was visible from added perylene. gave 1 and butyl benzoate (8) in addition to small amounts of the Norrish Type 11 products of 1. i.e. acetophenone (2) and cis- and trans- 2-methyl-1-phenylcyclobutanols. Biacetyl-sensitized photolysis of 5a in benzene yielded 2 in much higher yield in addition to 1, 8, and biphenyl. These results suggest that the triplet excited state of 1 is formed by the decomposition of 1 in low yield in thermolysis and in much higher yield in sensitized photolysis. although some of the Type II products may not arise from the triplet valerophenone.     

KINETICS OF ENZYMATIC PHOTOREACTIVATION STUDIED WITH PHAGE T1 AND ESCHERZCHZA COLI Bs-1*

Abstract— The kinetics of enzymatic photoreactivation (PR) of u.v.-induced killing was compared among E. coli B_s-1, phage T1 in B_s-1 and phage T1 in irradiated B_s-1. The PR action spectrum showed no substantial difference between PR of B_s-1 and PR of T1 in B_s-1. The PR D₃₇ (i.e. the PR dose required to reactivate all but 37 per cent of the reactivable lethal lesions) was found to decrease linearly with decreasing U.V. dose whether U.V. was given to produce pyrimidine dimers in B_s-1 DNA, which then compete with irradiated T1 DNA for PR enzyme, or to B_s-1 or T1 DNA to produce dimers serving as substrate for the PR enzyme. A generalized Michaelis-Menten formula was used to analyze the data and the following conclusions were drawn. (1) The number of PR enzyme molecules per cell available for PR of T1 DNA inside the B_s-1 host is only a quarter of the number available for PR of the B_s-1 host itself. (2) The Michaelis constant K_m for reaction of host-DNA-damage and PR-enzyme becomes larger when the host damage acts as competitive inhibitor to PR of T1 DNA than when it is the substrate for PR enzyme. (3) PR enzyme retains almost all its initial catalytic efficiency even after about two-hundred rounds of catalytic functioning. Conclusions (1) and (2) suggest that PR enzyme is concentrated within the nuclear area surrounding the host DNA.     

INACTIVATION OF PHAGE BY NEAR-ULTRAVIOLET RADIATION AND HYDROGEN PEROXIDE

A series of phage with different genomes (both single-stranded and double-stranded RNA and DNA) was inactivated with hydrogen peroxide (H₂O₂) in various combinations with far-ultraviolet (FUV) and near-ultraviolet (NUV) radiations. In every case but one (a lipid-coated phage), a sublethal H₂O₂ concentration greatly enhanced killing by NUV but not FUV. Moreover, this NUV/H₂O₂ synergism was oxygen independent and there was little if any host cell reactivation upon NUV plus H₂O₂ inactivation. These results suggest that these phage are inactivated by a common mechanism irrespective of nucleic acid composition, but that some phage genomes may be more vulnerable to NUV/H₂O₂ inactivation than others.     

ACTION SPECTRUM FOR INACTIVATION OF THE INFECTIVITY OF POTATO VIRUS X BY U.V. RADIATION

Abstract— Quantum yields for inactivation of infectivity of potato virus X by monochromatic ultraviolet radiation of wavelengths ranging from 230 to 290 nm, were measured with reference to energy absorbed by (a) the whole virus and (b) the virus RNA. The yields depended on the wavelength, but those with reference to energy absorbed by the RNA varied much less (with extreme values of 10^-3 and 1.9 ± 10^-3 than those with reference to whole virus. Consequently the action spectrum for inactivation of a dilute solution of the virus resembled the shape of the absorption spectrum of the RNA, but not closely enough to allow coincidence by adjusting the scales. The amount of photoreactivation increased as the wavelength increased and also as the year progressed from May to July; the extreme values of the photoreactivable sector were 0.43 and 0.86.     

PHOTOREACTIVATION ACTION SPECTRUM OF TOBACCO MOSAIC VIRUS-RIBONUCLEIC ACID INACTIVATED BY U.V. RADIATION*

Abstract— Initially photoreactivation of irradiated (2537 Å) nucleic acid on pinto bean increases linearly with time of illumination with white light of 250 ft-c. Maximum amounts of photo-reactivation depend on the quality of light used. The action spectrum shows a peak in the ‘black light’ region, where the greater amount of photoreactivation is found, and a shoulder in the blue light region. Maximum repair is obtained with ‘black light.’ Photoreactivation does not occur at wavelengths above 550 nm. Photoprotection by illumination of leaves prior to inoculation by irradiated RNA was not found. The action spectrum for photoreactivation does not resemble the action spectrum for photosynthesis.     

SUPPRESSION OF CONTACT HYPERSENSITIVITY BY UV RADIATION AND ITS RELATIONSHIP TO UV-INDUCED SUPPRESSION OF TUMOR IMMUNITY

Abstract— In this study, we examine some of the photobiologic and immunologic characteristics of the suppression of contact hypersensitivity (CHS) by UV radiation. BALB/c mice were irradiated on the shaved dorsal skin with FS40 sunlamps and sensitized 5 days later by applying a contact sensitizer lo the shaved abdomen. The suppression of CHS resulting from exposure to a given total dose of UV radiation was unaffected by changes in dose fractionation over a 5-day period and by changes in dose-rate over a 10-fold range. Elimination of wavelengths below 315 nm with a mylar filter abrogated the suppressive effect of the sunlamps, even when the same total energy was administered. Irradiation of unshaved mice required 14 times more energy to produce 50% suppression than was required for shaved mice, suggesting that the exposed skin is the primary target of this effect. Contact sensitization of UV-irradiated, but not unirradiated, mice induced the appearance of antigen-specific suppressor T lymphocytes in their spleen. The photobiologic and immunologic similarities between the suppression of CHS by UV radiation and the UV-mediated suppression of tumor rejection that we described previously suggest that these two immunosuppressive effects of UV exposure share certain steps in their pathways.     

聚(苯乙烯-甲基丙烯酸甲酯-砜)的合成及其紫外光降解

本文研究了苯乙烯、甲基丙烯酸甲酯、二氧化硫三元共聚合反应。其产品的特性用元素分析,红外和紫外光谱,NMR,DSC,TG等进行了详细地表征。该产品只有一个玻璃化温度132.5℃,故认为它是一个共聚物——聚(苯乙烯-甲基丙烯酸甲酯-砜),而不是一个均聚物的混合物。将聚(苯乙烯-砜)、聚(苯乙烯-甲基丙烯酸甲酯-砜)、聚甲基丙烯酸甲酯分别加热时,它们分别在252℃、272℃、282℃开始分解。当这些产品的溶液用三支30瓦的低压汞灯辐照时,聚(苯乙烯-砜)溶液的粘度下降速度比聚(苯乙烯-甲基丙烯酸甲酯-砜)快,聚甲基丙烯酸甲酯最慢。     

Liquid-holding effects in U.V.-irradiated phage T3

Abstract— Holding complexes of u.v.-irradiated (254 nm) T3 phage in E. coli B/r cells for several hours at 37°C in buffer, or broth with chloramphenicol, affects the phage survival in at least two different ways: (1) by enhancing excision repair, resulting under certain conditions in liquid-holding recovery (LHR), and (2) by destroying the phage (holding inactivation). LHR is most apparent in buffer containing 20 μg ml^-1 chloramphenicol (CAP). It is expressed by as much as a 10–fold increase in the fraction of complexes that display host-cell reactivation (resulting from excision repair), but the percentage of u.v. lesions repaired within repair-proficient complexes is slightly decreased. LHR is not observed if T3 infects the repair-deficient strain B_s-1. Holding inactivation is readily observed with unirradiated phage complexes in broth containing CAP. The response of irradiated-phage complexes to liquid-holding conditions is more complex: holding inactivation is less effective for irradiated than for unirradiated phage DNA (i.e. the irradiated DNA is to some extent ‘protected’), and processes leading to LHR are superimposed. Thus under certain holding conditions one observes the paradoxical phenomenon that the viable titer of irradiated phage is several times higher than that of unirradiated phage. The nature of holding inactivation is not known, nor is the mechanism by which irradiated DNA is partially protected against it. Holding inactivation does not require protein synthesis; it is rather enhanced at high CAP concentration and seems to be favored by otherwise active cell metabolism. At high CAP concentrations (200–400 μg ml^-1, as compared to 20 μg ml^-1) irradiated-phage complexes show neither LHR nor protection against holding inactivation. Likewise they fail to undergo some step by which the phage DNA becomes insensitive to repair inhibition by caffeine.     

METABOLISM OF NUCLEIC ACIDS IN U.V. IRRADIATED L-STRAIN CELLS TREATED WITH NATIVE DEOXYRIBONUCLEIC ACID

Abstract— In our earlier papers we described the recovery of U.V. irradiated L-strain cells by means of nucleic acids. In an attempt to get better insight in the mechanism(s) leading to this recovery we studied the effect of native DNA on the metabolism of nucleic acids of recipient cells. In these experiments we have found that the label isologous DNA-C¹⁴ is incorporated in DNA of normal and irradiated recipient cells. The rate of DNA-C¹⁴ label incorporation in the DNA of irradiated recipient cells is slower than in the unirradiated controls. The presence of thymidine in the incubation media reduces the incorporation rate of DNA-C¹⁴. When the irradiated cells have been growing in a medium containing isologous DNA in presence of thymidine or heterologous DNA the recovery effect was not observed.     

THE U.V. PHOTOCHEMISTRY OF CYTIDYLIC ACID

Abstract The ultraviolet (u.v.) irradiation of the 3' isomer of cytidylic acid (Cp) produces the hydrate (Cp*) with water added across the 5–6 double bond. The yield of this photo-product has been measured by, (a) separating the photoproduct by electrophoresis and (b) by observing the loss in absorbance. When corrections are made for reversal of the hydrate during the experiment, both methods gave the same result. Cross sections and quantum yields for the production of the hydrate were measured over the wavelength range 220 to 290 nm and over a pH range from 1 to 10. The quantum yield is markedly dependent on pH being higher by a factor of 6 to 10 for the neutral form. We have also demonstrated the existence of a very short lived photoproduct (half life 8–9 min) in both Cp3' and Cp5': The nature of this short lived product is not known.