On the use of ultraviolet light to study protein-DNA crosslinking.

Irradiation of Ehrlich ascites chromatin with ultraviolet light (u.v.) leads to protein-DNA crosslinking as determined by CsCl isopycnic ultra-centrifugation and SDS-polyacrylamide gel electrophoresis. At the most 4.5% of the chromatin proteins labelled with (14C)-lysine and (14C)-arginine can be crosslinked to DNA at u.v. doses between 3.6 X 10(4) to 10.8 X 10(4) ergs/mm2. We find however that the crosslinking reaction is hindered by protein breakage induced by u.v. light of wave length of less than 2950 A. Our results point out that caution must be used in the interpretation of studies on protein-nucleic acid interactions using u.v. light.     

Studies in experimental animal models of the effect of ultraviolet radiation (UVC) on blood and isolated cell populations

The cellular and molecular basis of the therapeutically used effect of reinjected ultraviolet ( UVC ) irradiated blood is unknown. First approaches to that problem were made in this study by aid of model experiments. Neither the spontaneous degranulation nor the antigen-induced histamine release from rat connective tissue mast cells (in vivo) was influenced by the injection (i.v.) of ultraviolet irradiated blood or blood lymphocytes. By comparison of the effect of ultraviolet light on blood lymphocytes (number of dead cells, strength of chemoluminescence) after irradiation of the isolated cells and the unfractionated blood respectively, we could show that the strong light absorption within the blood sample prevents damage or functional alterations of the blood lymphocytes. The compound 48/80-induced histamine release from rat peritoneal mast cells can be completely inhibited by ultraviolet irradiation (0.6 mJ/cm2) without increasing the spontaneous histamine release.     

The Effect of Hydrogen Peroxide and Ultraviolet Irradiation on Non-sporing Bacteria

A kill of 99.99% was obtained in cell suspensions of Escherichia coli and Streptococcus faecalis by incubation with hydrogen peroxide 1.0% (w/v) for 75 and 180 min respectively. The same kill was produced by 30 s irradiation with ultraviolet (u.v.) light in the presence of hydrogen peroxide 1.0% (w/v). This simultaneous treatment with u.v. and hydrogen peroxide produced a synergistic kill at least 30-fold greater than that produced by irradiation of cell suspensions of Esch. coli with or without subsequent incubation with hydrogen peroxide.     

Pyrimidine dimers are not the principal pre-mutagenic lesions induced in lambda phage DNA by ultraviolet light

Experiments were performed to examine the role of cyclobutyl pyrimidine dimers in the process of mutagenesis by ultraviolet (u.v.) light. Lambda phage DNA was irradiated with u.v. and then incubated with an Escherichia coli photoreactivating enzyme, which monomerizes cyclobutyl pyrimidine dimers upon exposure to visible light. The photoreactivated DNA was packaged into lambda phage particles, which were used to infect E. coli uvr- host cells that had been induced for SOS functions by ultraviolet irradiation. Photoreactivation removed most toxic lesions from irradiated phage, but did not change the frequency of induction of mutations to the clear-plaque phenotype. This implies that cyclobutyl pyrimidine dimers can be lethal, but usually do not serve as sites of mutations in the phage. The DNA sequences of mutants derived from photoreactivated DNA showed that almost two-thirds (16/28) were transitions, the same fraction found for u.v. mutagenesis without photoreactivation. These results show that in this system, the lesion inducing transitions (the major type of u.v.-induced mutation) is not the cyclobutyl pyrimidine dimer; a strong candidate for a mutagenic lesion is the Pyr(6-4)Pyo photoproduct. On the other hand, photoreactivation of SOS-induced host cells before infection with u.v.-irradiated phage reduced mutagenesis substantially. In this case, photoreversal of cyclobutyl dimers serves to reduce expression of the SOS functions that are required in the process of targeted u.v. mutagenesis.     

Ultraviolet reactivation of lambda phage: assay of infectivity of DNA molecules by spheroplast transfection

Prior irradiation of non-lysogenic bacteria by ultraviolet light leads to an increase in the viability of infecting irradiated λ phage (ultraviolet reactivation). Similarly, u.v. irradiation of wild type or uvrD bacteria lysogenic for λcIind^? increased the fraction of closed circular duplex phage DNA molecules formed after infection with u.v.-irradiated λ phage. The closed circular molecules isolated from the irradiated lysogens were shown to be free from u.v. damage by a spheroplast transfection assay. The increase of closed circular molecules is sufficient to explain the ultraviolet reactivation observed by the increase of viability of irradiated phage.In ultraviolet reactivation, damage must be erased on irradiated DNA molecules and the repair is independent of total replication of phage genomes, exchange of sister chromatids or recombination between phage genomes. Protein synthesis is necessary to increase the level of closed circular molecules of irradiated λ phage after irradiation of bacteria.     

Role of DNA repair genes and an R plasmid in conferring cryoresistance on Pseudomonas aeruginosa

The resistance of Pseudomonas aeruginosa wild-type, uvr, pol and rec strains to ultraviolet (u.v.) light, X-rays and freezing and thawing was determined. An R plasmid, pPL1, which increased resistance of the wild-type uvr, and pol but not rec strains to u.v. light, increased the resistance of only rec and pol mutants to X-rays and freezing and thawing. These findings reinforce the idea of DNA as a target in the organism for freeze-thaw stress and suggest that freeze-thaw-induced DNA damage might be similar to that produced by X-rays but different from that produced by u.v. light.     

The destruction of spores of Bacillus subtilis by the combined effects of hydrogen peroxide and ultraviolet light

Ultraviolet light irradiation of bacterial spores in the presence of hydrogen peroxide has been shown to produce synergistic kills when compared with ultraviolet light (u.v.) and hydrogen peroxide used sequentially. This use in combination has been patented for the commercial sterilization of packaging before filling with UHT-processed products. Previous results have shown that lamps producing u.v. light with a maximum output at about 254 nm were extremely effective. Results obtained using a Synchrotron radiation source to produce a narrow band of irradiation now shows that the greatest kill of spores of Bacillus subtilis in the presence of hydrogen peroxide is obtained with radiation at ˜270 nm. Such results suggest that the action of the u.v. light is not directly on the spore DNA but may be related to the production of free hydroxyl radicals from hydrogen peroxide.     

Inactivation of Escherichia coli by ultraviolet light and hydrogen peroxide in a thin film contactor

Apparatus for irradiating enclosed thin liquid films with ultraviolet (u.v.) light (Λ= 253.7 nm) in combination with hydrogen peroxide was used to inactivate Escherichia coli in water. Hydrogen peroxide concentrations of 2.5, 5.0 and 10.0 g/I were used and in each case synergistic inactivation was observed. At the highest concentration, a fractional survival of 1.3 times 10^-3 was obtained after 20 min; this was decreased to 3.1 times 10^-6 by simultaneous u.v. irradiation.     

The kinetics of Bacillus subtilis spore inactivation on filter paper by u.v. light and u.v. light in combination with hydrogen peroxide

Inactivation of spores of Bacillus subtilis (ATCC 6633) on two different grades of cellulose filter paper (Whatman Grades 2 and 6), by ultraviolet light (u.v.), at an intensity of approximately 4·5 Wm⁻² and at fluences of up to 2 × 10³ Jm⁻², and u.v. in the presence of hydrogen peroxide, is described in terms of multi-target and single hit–single target kinetic expressions. Wet spores were inactivated at rates ranging from 6·7 to 10·6 higher than that of dry spores on both grades of filter paper. In addition, spore inactivation was up to 5·6 times more rapid on Grade 2 filter paper. Synergistic inactivation was seen to occur when spores were irradiated in the presence of 1% (w/v) hydrogen peroxide with rates up to 5·3 times higher than with treatment solely by u.v. The results obtained are discussed in general terms with particular reference to surface characteristics which might provide shielding to micro-organisms from incident u.v. light.     

A cell kinetic model to explain the time of appearance of skin reaction after X-rays or ultraviolet light irradiation.

Skin reactions to various doses of X-rays (300 and 10 kV) and ultraviolet light (u.v.) have been compared using hairless mice. Two regions of epidermis with widely differing cell kinetics and gross structure have been compared. Little evidence could be found to support the idea that the early phases of the reaction are dependent on cell cycle time. The data can be explained by a model based on the assumption that epidermis contains only a small fraction of clonogenic (stem) cells and this fraction may vary in different epidermal regions. X-rays appear to exert their greatest destructive action on these clonogenic cells while u.v. is more indiscriminate in its action, killing both clonogenic and non-clonogenic cells.     

Sporulation of Botrytis cinerea as affected by photoselective polyethylene sheets and filters

Sporulation of Botrytis cinerea was studied in vitro under various filters and polyethylene co-polymer (PE) sheets. It was found that continuous darkness blocked sporulation completely. Sporulation was inhibited by reduced ultraviolet (u.v.) radiation. Almost total inhibition of sporulation was also found when cultures were grown under a blue filter. The reduction of spore formation was negatively related to the ratio of transmitted blue/u.v. light. The possibility of using this effect to control the incidence of Botrytis cinerea in greenhouses is discussed. A brief report has been published (Reuveni, Raviv, Allingham & Bar, 1988).     

The Influence of the Environment and Clothing on Human Exposure to Ultraviolet Light

Objection

The aim of this study is to determine the effect of clothing and the environment on human exposure to ultraviolet light.

Methods

The ultraviolet (ultraviolet A and ultraviolet B) light intensity was measured, and air quality parameters were recorded in 2014 in Beijing, China. Three types of clothing (white polyester cloth, pure cotton white T-shirt, and pure cotton black T-shirt) were individually placed on a mannequin. The ultraviolet (ultraviolet A and ultraviolet B) light intensities were measured above and beneath each article of clothing, and the percentage of ultraviolet light transmission through the clothing was calculated.

Results

(1) The ultraviolet light transmission was significantly higher through white cloth than through black cloth; the transmission was significantly higher through polyester cloth than through cotton. (2) The weather significantly influenced ultraviolet light transmission through white polyester cloth; transmission was highest on clear days and lowest on overcast days (ultraviolet A: P=0.000; ultraviolet B: P=0.008). (3) Air quality parameters (air quality index and particulate matter 2.5 and 10) were inversely related to the ultraviolet light intensity that reached the earth’s surface. Ultraviolet B transmission through white polyester cloth was greater under conditions of low air pollution compared with high air pollution.

Conclusion

Clothing color and material and different types of weather affected ultraviolet light transmission; for one particular cloth, the transmission decreased with increasing air pollution.     

A CELL KINETIC MODEL TO EXPLAIN THE TIME OF APPEARANCE OF SKIN REACTION AFTER X-RAYS OR ULTRAVIOLET LIGHT IRRADIATION

Skin reactions to various doses of X-rays (300 and 10 kV) and ultraviolet light (u.v.) have been compared using hairless mice. Two regions of epidermis with widely differing cell kinetics and gross structure have been compared. Little evidence could be found to support the idea that the early phases of the reaction are dependent on cell cycle time. The data can be explained by a model based on the assumption that epidermis contains only a small fraction of clonogenic (stem) cells and this fraction may vary in different epidermal regions. X-rays appear to exert their greatest destructive action on these clonogenic cells while u.v. is more indiscriminate in its action, killing both clonogenic and non-clonogenic cells.     

Low pressure ultraviolet studies for inactivation of Giardia muris cysts

AIMS: The research was initiated to confirm earlier ultraviolet (u.v.) light inactivation studies performed on Giardia cysts using excystation as the viability indicator. Following this, a comparison of in vitro excystation and animal infectivity was performed for assessing cyst viability after exposure to low-pressure u.v. irradiation. METHODS AND RESULTS: Cysts of Giardia muris were inactivated using a low-pressure u.v. light source. Giardia muris was employed as a surrogate for the human pathogen Giardia lamblia. Cyst viability was determined by both in vitro excystation and animal infectivity. Cyst doses were counted using a flow cytometer for the animal infectivity experiments. Using in vitro excystation as the viability indicator, fluences as high as approximately 200 mJ cm(-2) did not prevent some cysts from excysting, thus verifying earlier work. Using animal infectivity, u.v. fluences of 1.4, 1.9 and 2.3 mJ cm(-2) yielded log10 reductions ranging from 0.3 to >or= 4.4. CONCLUSIONS: Results indicate that in vitro excystation is not a reliable indicator of G. muris cyst viability after u.v. disinfection. Very low doses of u.v. light rendered G. muris cysts non-infective in the mouse model employed. SIGNIFICANCE AND IMPACT OF THE STUDY: Data presented represent the only complete u.v. inactivation curve for G. muris. This research provides evidence that u.v. can be an effective barrier against Giardia spp. in the treatment of drinking water supplies.     

Separation of adreno-ovarian steroids by thin-layer chromatography

The major adreno-ovarian steroid hormones (progesterone, estrone, 17α-estradiol, 17β-estradiol, estriol, corticosterone, cortisone, and cortisol) have been separated simultaneously on a single TLC plate without recourse to transfer chromatography. The plate was developed successively twice in benzene/ethanol (95:5, v/v) solvent system. It was then sprayed with rhodamine 6G and a line was drawn isolating the already separated least polar and medium-polar steroids (progesterone, estrone, 17α-estradiol, and 17β-estradiol) with the help of ultraviolet light. Then 5 ml methanol per 100 ml solvent in the tank was added and the plate again developed 2–3 times up to the line drawn, when polar steroids (corticosterone, cortisone, cortisol, and estriol) separated out.     

Resistance of Serratia marcescens to Hydrogen Peroxide

Irradiation with ultraviolet (u.v.) light (71 J/m²) reduced the viable count of suspenrsions of Serratia marcescens , grown in a glycerol-salts defined medium, to five in 10⁴ cells. Subsequent incubation of irradiated cells in hydrogen peroxide failed to decrease the survivors, but u.v. irradiation in the presence of hydrogen peroxide reduced the viable count to fewer than two in 10⁶ cells. Cells grown in defined medium with added iron had more measurable catalase activity and were more resistant to hydrogen peroxide alone and to simultaneous treatment with u.v. irradiation and hydrogen peroxide. Cells grown in a non-defined medium contained little iron and measurable catalase activity but were more resistant to hydrogen peroxide. Treatment with toluene, heat killing or sonication increased the catalase activity detected in all cell suspensions and showed that resistance to hydrogen peroxide and to u.v. irradiation in hydrogen peroxide was related to the total catalase activity within cells.     

Recovery from UV-induced potentially lethal damage in systemic lupus erythematosus skin fibroblasts

The repair of ultraviolet light-induced potentially lethal damage was investigated in density-inhibited skin fibroblast cell strains derived from patients with systemic lupus erythematosus. The effect of exposure to polychromatic ultraviolet light composed of environmentally relevant wavelengths or to the more commonly studied, short wavelength (254 nm) ultraviolet light was studied. Systemic lupus erythematosus cells, which are hypersensitive to ultraviolet light under growth promoting conditions, were able to repair potentially lethal damage as well as normal cells.     

The composition and structure of walls of dark fungus spores

Summary Dark pigmented fungi predominate in desert soils. The dark pigment of the spores is a melanin. These dark spores resist ultraviolet radiation of 2537 Å. The degree of opacity depends on the amount and location of the melanin.In the presence of hexachloracetone, about half of the spores produced in culture are light colored, and easily killed by u.v. light. Electronmicrographs are presented showing spore wall structure of several representative fungi at high resolution.     

Ultraviolet light activates blocking actions of dantrolene on intracellular Ca2+ release in bullfrog sympathetic neurones

Effects of dantrolene, a blocker of intracellular Ca2+ release, on the oscillation of the intracellular Ca2+ ([Ca2+]i) induced by caffeine were studied in bullfrog sympathetic ganglion cells, using a Fura-2 fluorescence technique. Dantrolene blocked the Ca2+ oscillation only in the cell illuminated by ultraviolet light (335-385 nm). Likewise, the blocking effects on rhythmic Ca(2+)-dependent hyperpolarizations, representing Ca2+ oscillations via activation of Ca(2+)-dependent K+ channel, occurred only under the illumination with ultraviolet light (335-385 nm), but not with visible light (404-417 nm). This wavelength dependence differs from the absorbance spectrum of dantrolene. On the other hand, dantrolene preirradiated with ultraviolet light under dark condition or ultraviolet light itself did not affect the [Ca2+]i oscillation. The blocking action was not prevented by the pretreatment of the cells with reducing agents. These results indicate that illumination of the Ca2+ release channel or dantrolene itself with ultraviolet light (possibly the former) is necessary for the drug to exert its blocking effect. Furthermore, dantrolene was found to decrease Fura-2 fluorescence and to increase cell autofluorescence, leading sometimes to a false decrease in the basal [Ca2+]i.     

Mechanisms of photochemotherapy-induced apoptotic cell death in lymphoid cells.

Previous studies we performed showed that 8-methoxypsoralen in combination with ultraviolet A light (photochemotherapy) caused DNA damage and that this caused nucleotide depletion in peripheral blood leukocytes, secondary to an active form of programmed cell death, poly(ADP-ribosyl)ation. Further studies revealed that 24 h after exposure to 10 J/cm2 ultraviolet A light and 8-methoxypsoralen (300 ng/mL), apoptotic cells increased from 3 (control) to 31% (p less than 0.001). Ultraviolet A light alone also significantly increased the number of apoptotic cells. These morphological changes were confirmed by parallel findings on DNA electrophoresis. Treatment with 2 to 5 J/cm2 of ultraviolet A light and 8-methoxypsoralen caused an approximately 30% increase in cytosolic free calcium levels in peripheral blood leukocytes 1 h after exposure. Associated with this was a 51% increase in 45Ca2+ uptake over the first 60 min. Similar findings in a different lymphoid cell (CCRF-CEM) confirmed the results obtained with peripheral blood leukocytes. The use of calcium-free medium prevented a rise in cytosolic free calcium and decreased the number of cells undergoing apoptotic cell death. Cycloheximide inhibited ultraviolet A light - 8-methoxypsoralen induced apoptosis in CCRF-CEM cells; it also decreased calcium levels in control CCRF-CEM cells. This study shows that ultraviolet A light - 8-methoxypsoralen caused apoptotic cell death in lymphoid cells; this appeared to be associated with calcium influx, presumably because of the requirement of endogenous endonucleases for calcium.