THE ISOLATION AND PARTIAL CHARACTERIZATION OF UV SENSITIVE MUTANTS OF Synechococcus PCC7943 (Anacystis nidulans 602)

Abstract –Four mitomycin-C sensitive, UV sensitive mutants of Synechococcus PCC7943 were isolated. They were capable of host-cell reactivation and showed enhanced UV survival if held under non-growth conditions for a recovery period immediately after irradiation. One of the mutants was shown to be deficient in Weigle-reactivation.     

REPAIR OF NEAR (365 nm)- AND FAR (254 nm)-UV DAMAGE TO BACTERIOPHAGE OF ESCHERICHIA COLI

Abstract: Intact bacteriophage have been irradiated at 365 nm or at 254 nm and then analysed for DNA photoproducts or injected into their bacterial host to test susceptibility of the damage to both phage and host-cell mediated repair systems. Both thymine dimers and single-strand breaks are induced in the phage DNA by 365 nm radiation. The dimers appear to be the major lethal lesion (approximately 2 dimers per lethal event) in both repair deficient bacteriophage T4 and bacteriophage λ. after irradiation with either 254 nm or 365 nm radiation. Damage induced in T4 by either wavelength is equally susceptible to x -gene reactivation (repair sector approximately 0.5). v -gene reactivation acts on a larger fraction of the near-UV damage (repair sector of 0.82 at 365 nm as against 0.66 at 254 nm). The host-cell mediated photoreactivation system is only slightly less effective for near-UV damage but host-cell reactivation (as measured by comparing survival of phage λ. on a uvr⁺ and a uvr^- host) is effective against a far smaller sector of near-UV damage (0.35) than far-UV damage (0.85). Weigle-reactivation (far-UV induced) of near-UV damage to phage λ is not observed. The results suggest that unless the near-UV damaged phage DNA is repaired immediately after injection. the lesions rapidly lose their susceptibility to repair with a consequent loss of activity of the phage particles.     

UV-LYSOGENIC INDUCTION OF Λ PHAGE IN lexA1 MUTANTS OF Escherichia coli: KINETICS OF THE PROCESS

Abstract— Although the lex gene has been described until recently as being required for lysogenic induction, both our work and the work of others have reported Λ prophage induction in some lexA mutants. However, the characteristics of the process were not defined. We describe UV induction of prophage in a lexA1 mutant at a slightly lower level and requiring 2 times longer than the wild type. As demonstrated in some work, in cells treated with low levels of rifampicin (RIF) no new synthesis of RecA protein is needed for the prophage induction although the onset of lysis is delayed. We suggest that the lysogenic induction in lexA cells is due to the same mechanism that induces prophage in the wild type cells treated with RIF. That is, the induction is due to the cleavage of Λ represser by the basal RecA protease in the DNA-single-strand gap, since RecA protease and monomer represser both have high affinity for this type of DNA. So, LexA protein need not be cleaved for the prophage induction.
No Weigle-reactivation (WR) was detected in the lex mutant even after a long post-irradiation incubation, suggesting that unlike prophage induction, WR requires LexA protein cleavage.     

POLYCAPROLACTAM MODIFIED BY POLYPHENYLQUINOXALINE

Three polycaprolactam samples modified by 0.05—0.50% polyphenylquinoxaline(PPQ)by weight were prepared.Their structure and mechanical properties were characterized by FT-IR, SEM, density, tensile, impact, DTAand visco-elastic method, PPQ induced the formation of "crystal grains" distributed evenly over nylon spherulites in modified samples, which were observed for the first time, and strengthened modified samples. Modified nylons had higher crystallinities, higher T_g, more nearly perfect spherulites than MC nylon itself, and showed typical reinforcing effect on mechanical properties.     

ACTIVATION OF NF-KB IN HUMAN SKIN FIBROBLASTS BY THE OXIDATIVE STRESS GENERATED BY UVA RADIATION

We have examined the role of the nucleus and the membrane in the activation of nuclear factor (NF)-KB by oxidant stress generated via the UVA (320–380nm) component of solar radiation. Nuclear extracts from human skin fibroblasts that had been irradiated with UVA at doses that caused little DNA damage contained activated NF-KB that bound to its recognition sequence in DNA. The UVA radiation-dependent activation of NF-KB in enucleated cells confirmed that the nucleus was not involved. On the other hand, UVA radiation-dependent activation of NF-KB appeared to be correlated with membrane damage, and activation could be prevented by a-tocopherol and butylated hydroxytol-uene, agents that inhibited UVA radiation-dependent peroxidation of cell membrane lipids. The activation of NF-KB by the DNA damaging agents UVC (200–290nm) and UVB (290–320nm) radiation also only occurred at doses where significant membrane damage was induced, and, overall, activation was not correlated with the relative levels of DNA damage induced by UVC/UVB and UVA radiations. We conclude that the oxidative modification of membrane components may be an important factor to consider in the UV radiation-dependent activation of NF-KB over all wavelength ranges examined.     

单分子膜诱导下的矿物晶体生长

介绍了近几年单分子膜诱导下矿物晶体生长的最新进展。讨论了单分子膜诱导下的矿物晶体生长与溶溶中的异同,成膜材料,单分子膜性质和聚集态,矿物晶体的选择性及晶体的表征方法。     

INDUCTION OF phr GENE EXPRESSION BY PYRIMIDINE DIMERS IN Escherichia coli

The photoreactivating enzyme (PRE) is concerned with mainly two kinds of light wavelength. The PRE splits UVC (254 nm)-induced pyrimidine dimer by absorbing UVA (320–380 nm) or visible light in its chromophore. The present paper demonstrates that the phr gene expression was efficiently induced in an excision defective strain (uvrA∼) after irradiation by UVC and UVB (290-320 nm), but not by UVA and visible light. In addition, the induced activity was significantly depressed by irradiation with UVA and visible light. Therefore we conclude that the phr gene expression can be induced by pyrimidine dimers.     

A NOVEL REDUCTIVE CYCLIZATION OF ARYLMETHYLIDENEMALONONITRILE PROMOTED BY LOW-VALENT TITANIUM

ABSTRACT

The intermolecular and intramolecular reductive coupling reactions of arylmethylidenemalononitriles induced by low-valent titanium have been studied. A possible reaction mechanism is proposed.     

PHOTOSENSITIZATION BY METHOTREXATE PHOTOPRODUCTS

Abstract— Photoproducts induced upon excitation of methotrexate by UV light have been separated by ion exchange chromatography. They include 2,4-diamino-6-pteridinecarboxylic acid, 2,4-diamino-6-pteridine-carboxaldehyde and other unidentified pteridine derivatives. The same photoproducts can be also formed upon photodynamic reaction using hematoporphyrin as photosensitizer. In oxygen saturated aqueous solutions (pH∼7), methotrexate photoproducts sensitize the oxidation of histidine and tryptophan by UV light by a process involving singlet oxygen. In aqueous solutions containing albumin or in human serum, the same photoproducts are formed from free methotrexate but not from albumin-bound methotrexate. In the latter case the results may suggest that methotrexate covalently binds to albumin upon excitation with UV light either in absence or in presence of oxygen. These results could explain the photosensitization accompanying cancer chemotherapy with high dose methotrexate and also the synergistic effects of PUVA + low dose methotrexate in psoriasis therapy.     

INDUCED REPAIR IN ESCHERICHIA COLI: INDUCTION BY ULTRAVIOLET LIGHT AT-79°C

Two of the phenomena associated with induced (S.O.S.) repair, namely induced inhibition of post radiation DNA degradation and induced radioresistance have been shown to be elicited by 245 nm radiation applied to E. coli cells in the frozen state at -79°C. The effect of radiation in this condition is considerably less photoreactivable than similar effects produced by exposure in the wet state. Since protein-DNA crosslinks are believed to be formed under these conditions, such consequences of UV radiation appear to be a potent inducer of induced repair.     

POTENTIATION OF OXIDATIVE DAMAGE TO PROTEINS BY ULTRAVIOLET-A AND PROTECTION BY ANTIOXIDANTS

We have studied the damage of alcohol dehydrogenase (ADH) and glyceraldehyde 3-phosphate dehydrogenase (GAPD) induced by Fe++/EDTA + H2O2 in combination with UV-A (main output at 365 nm). Enzyme inactivation, formation of hydroxyl radicals (measured in the absence of enzymes), increase in protein carbonyls, oxidation of sulfhydryl (SH) groups, loss of native protein fluorescence, and enhanced protease degradation were used to determine protein damage. Hydroxyl radical production was greatly enhanced by the combination of UV-A with Fe++/EDTA + H2O2. The combined treatment increased protein carbonyls but decreased native protein fluorescence and SH groups. The combined treatment caused turbidity in GAPD but not in ADH, whereas trypsin susceptibility was increased more in ADH than in GAPD. These measurements of protein oxidation correlated well with enzyme activities. Glyceraldehyde 3-phosphate dehydrogenase and dithiothreitol were most protective against such damage, while hydroxyl radical and singlet oxygen scavengers were partially effective. Superoxide dismutase had no effect. Thus, UV-A potentiation of protein damage induced by FE++/EDTA + H2O2 appeared to involve hydroxyl radicals and perhaps singlet oxygen but not superoxide radicals. The damage to proteins induced by combination of UV-A with physiological oxidants, iron ions and H2O2 may be relevant to UV-A-induced skin and tissue damage.     

LASER PHOTOSENSITIZATION OF CELLS BY HYPERICIN

Abstract— Administering a light dose of 90 J/cm² at 599 nm during incubation with hypericin to a highly differentiated normal epithelial cell line(FRTL–5), derived from Fisher rat thyroid, and to a neoplastic cell line(MPTK–6), derived from the lung metastases of a thyroid carcinoma induced in Fisher rats, produces cell kill at drug doses 1000 times lower than those necessary to cause the same mortality in the dark. The photocytocidal activity of this polycyclic quinone drug on neoplastic cells is superior to that of antitumor anthraquinone drugs, such as daunomycin and mitoxanthrone, and to the photosensitized antiviral activity previously reported for hypericin.     

DAMAGE OF SILICONE RUBBER INDUCED BY PROTON IRRADIATION

In this paper, the damage to methyl silicone rubber induced by irradiation with protons of 150 keV energy wasstudied. The surface morphology, tensile strength, Shore hardness, cross-linking density and glass transition temperaturewere examined. Positron annihilation lifetime spectrum analysis (PALS) was perfomed to reveal the damage mechanisms ofthe rubber. The results showed that tensile strength and Shore hardness of the rubber increased first and then decreased withincreasing irradiation fluence. The PALS characteristics τ_3 and I_3, as well as the free volume V_f, decreased with increasingirradiation fluence up to 10~(15) cm~(-2), and then increased slowly. It indicates that proton irradiation causes a decrease of freevolume in the methyl silicone rubber when the fluence is less than 10~(15)cm~(-2), while the free volume increases when thefluence is greater than 10~(15)cm~(-2). The results on cross-linking density indicate that the cross-linking induced by protonirradiation is dominant at smaller proton fluences, increasing the tensile strength and Shore hardness of the rubber, while thedegradation of rubber dominates at greater fluence, leading to a decrease of tensile strength and Shore hardness.     

FLUORESCENCE QUENCHING OF INDOLE BY DIMETHYLFORMAMIDE

Abstract— The effect of dimethylformamide on the lifetime (T) and quantum yield ( F ) of indole has been determined. Negative Stern-Volmer deviations prevail for T and F , but F ⁰/ F T⁰/T at all DMF concentrations. The deviation between F ⁰/ F and T⁰/T is ascribed to changes in F ⁰ and T⁰ induced by the change in solvent composition at the higher quencher concentrations.     

SYNTHESIS OF 3H-PYRROLES PROMOTED BY SAMARIUM/CAT. IODINE SYSTEM

ABSTRACT

The intermolecular reductive coupling of 1,1-diaryl-2,2-dicyanoethylenes or 1,1-diaryl-2-cyano-2-ethoxycarbonylethylenes with aromatic nitriles induced by samarium and catalytic iodine was studied. 3H-Pyrroles were prepared in moderate to good yields under neutral and mild conditions.     

ENHANCEMENT OF ULTRAVIOLET OR N-METHYL-N'- NITRO-N-NITROSOGUANIDINE SENSITIVITY OF DICTYOSTELIUM DISCOIDEUM BY 3-AMINOBENZAMIDE

Abstract— The amoeboid cells of Dktyostelium discoideum NC–4 possess a 3-aminobenzamide(3-ABA)-sensitive repair mechanism for DNA damages induced by UV-irradiation or N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-treatment. We have studied the effect of 3-ABA on each step of excision repair in the UV- irradiated cells. Although the nicking of DNA-strand and the excision of pyrimidine dimcrs are insensitive to 3-ABA, the rejoining of DNA strand-breaks is sensitive. The frequency of mutation induced by UV-irradiation or MNNG-treatment is depressed by 3-ABA. The mechanisms of repair inhibition by 3-ABA are discussed.     

THE DESTRUCTION OF TRYPTOPHANYL RESIDUES IN TRYPSIN BY 280-nm RADIATION

Abstract— The destruction of tryptophanyl residues in trypsin by 280-nm radiation was studied in relation to enzyme inactivation. Quantum yields for destruction of this residue (determined using the pDAB reagent) and for the inactivation of trypsin were measured when the enzyme was exposed to different environmental perturbations. The conformational modifications of trypsin induced in 6 M guanidine-HCl did not alter the rates of tryptophan destruction and enzyme inactivation. However, an enhanced destruction of the tryptophanyl residues was observed when trypsin solutions were irradiated at 60°C in the presence of air. The increased rate of tryptophan destruction at this temperature was not accompanied by a corresponding increase in the inactivation quantum yield. It was concluded that any photochemically induced reactions of this chromophore that are sensitive to conformational modifications or that result in the destruction of the indole ring are not important in the inactivation mechanism of trypsin.     

DESTRUCTION OF PHOTOREACTIVATING ENZYME BY 365 nm RADIATION*

Abstract— Following the observation that in vivo photoreactivation of 365-nm-induced pyrimidine dimers could not be observed chemically, a study was made of the inactivation of photoreactivating enzyme activity by this near-ultraviolet wavelength. It was observed that: (1) Dimers induced in extracted bacterial DNA by 365 nm radiation are completely photoreactivable and are monomerized as an exponential function of the photoreactivation time. (2) Photoreactivability of 254-nm-induced damage in Escherichia coli B/r Hcr is progressively destroyed in vivo as a function of the dose of 365 nm radiation. (3) The ability of the yeast photoreactivating enzyme to monomerize dimers induced at 365 nm in bacterial DNA is destroyed in vitro as a function of the dose of 365 nm radiation, and at a rate comparable to killing of E. coli. These results are consistent with biological measurements which indicate that photoreactivability of ultraviolet (near and far) lethal damage is reduced by exposure of the bacteria to 365 nm radiation.     

PHOTOSENSITIZATION BY PORPHYRINS DELIVERED TO L CELL FIBROBLASTS BY HUMAN SERUM LOW DENSITY LIPOPROTEINS. A MICROSPECTROFLUOROMETRIC STUDY

Human serum LDL were used as vehicles to deliver protoporphyrin and hematoporphyrin dimer to L cell mouse fibroblasts. Topographic analysis by microspectrofluorometry on single living cells shows that after digestion of LDL, protoporphyrin is localized in cytoplasmic areas. Protoporphyrin and hematoporphyrin dimer are readily bleached by 420 ± 60 nm radiations at the high fluence rate used. Complex bleaching kinetics are observed. Spectral studies using the same technique demonstrate that an intense fluorescent emission (λ_max= 450 nm) is produced immediately after the onset of irradiation with 365 ± 2 nm or 420 ± 60 nm radiations using LDL loaded with protoporphyrin or Photofrin II. These fluorescent products have been previously identified as lipofuscin-like pigments formed by reaction of lipid photoperoxides with amino groups. The permeation of lysosomal membranes is also induced after delivery of the porphyrins by LDL. This permeation can be strongly inhibited not only by the lysosomal inhibitors chloroquine and monensin but also by a-tocopherol. On the other hand, neither α-tocopherol nor chloroquine or monensin inhibit the lipofuscin-like pigment formation.     

PHOTOINACTIVATION OF CHINESE HAMSTER CELLS BY HEMATOPORPHYRIN DERIVATIVE AND RED LIGHT

Abstract— The use of hematoporphyrin derivative (HpD) has previously been demonstrated to be beneficial in clinical cancer therapy. This paper describes cell culture studies used to examine HpD phototherapy in Chinese hamster ovary cells (line CHO). Survival curves have been obtained for both direct HpD toxicity and HpD induced photoinactivation. Examination of HpD induced photoinactivation as a function of stage in the cell growth cycle has also been performed, as has the quantitative measurement of HpD uptake in cells (using ³H-HpD) as a function of cellular incubation time, serum concentration in the incubation medium, and cell cycle position. In the absence of light, no toxicity was observed for HpD incubation levels of up to 400 μg/m/ when incubations times were 3 h or less. Exposure of cells to light alone (> 590 nm, 4.0 mW/cm²) for 9 min was also found to be completely nontoxic. Survival curves obtained for exponentially growing cells labeled with various concentrations of HpD and subsequently illuminated with red light exhibited a threshold or shoulder region at short exposure times followed by exponential killing at longer exposure times. The cell cycle response curves for HpD induced photoinactivation of synchronized CHO cells was nearly flat, indicating no variation in sensitivity for cells treated at time periods from 6 to 15 h after mitosis. Additon of serum to the incubation medium resulted in improved plating efficiency and reproducible survival curves but decreased cellular uptake of HpD.