The fluence rate determines the synergistic interaction of UV radiation and heat for mitotic recombination and cell inactivation in yeasts

The significance of the UV fluence rate for the synergistic interaction of UV light (254 nm) and heat was demonstrated for the frequency of mitotic recombination in a wild-type diploid yeast of Saccharomyces cerevisiae (strain T1) and for cell inactivation of two wild-type diploid yeast of S. cerevisiae (strains T1, XS800). It was shown for mitotic recombination that a decrease in the intensity of UV exposure results in the necessity of decreasing the temperature at which UV irradiation occurs to provide the same value of the synergistic enhancement ratio. For cell inactivation, there was a specific temperature maximizing the synergistic effect for any constant fluence rate and the temperature range, synergistically increasing the inactivation effect of UV radiation, should be shifted to lower temperatures with a decrease in the fluence rate. To interpret the results observed, a simple mathematical model of the synergistic interaction was applied. The model suggests that the synergistic interaction of UV light and hyperthermia is expected to result from some additional effective damages arising from the interaction of some sublesions induced by both agents. On the basis of data obtained, it was supposed that the synergistic interaction of these factors might take place at small intensities of UV light and temperatures existing in the biosphere. In other words, for a long duration of interaction, which is important for problems of UV light protection and health effects, one can expect a synergistic interaction of this factor with environmental heat or physiological temperatures and thereby an increase in their inactivating and genetic consequences.     

Mathematical description of synergistic interaction of UV light and hyperthermia for yeast cells

A new mathematical model for the synergistic interaction of lesions produced by ultraviolet (UV) light and high temperature has been proposed. The model suggests that synergism is expected from the additional lethal lesions arising from the interaction of sublesions induced by both agents. These sublesions are considered noneffective after each agent is taken alone. The model predicts the dependence of the synergistic interaction on the ratio of lethal lesions produced by each agent applied, the greatest value of the synergistic effect as well as the conditions under which it can be achieved, and the dependence of synergistic effect on UV light fluence rate. These predictions of the model have been tested for the simultaneous combined action of UV light (wavelength 254 nm) and heat (45-57.5 degrees C) on two strains of wild-type diploid yeast cells of Saccharomyces cerevisiae. The theory appears to be appropriate and the conclusions valid.     

PHOTOCHEMISTRY OF THE BISBENZIMIDAZOLE DYE 33258 HOECHST WITH BROMODEOXYURIDINE AND ITS BIOLOGICAL EFFECTS ON BrdUrd-SUBSTITUTED ESCHERICHIA COLI

Abstract— Exposure of BrdUrd-substituted E. coli cells to 360 nm light in the presence of the bisbenzimi-dazole dye 33258 Hoechst increases their sensitivity dramatically. Mutant cells deficient in excision repair of DNA damage ( uur B) are more sensitive than wild type cells, indicating that the cells are able to repair this type of damage. However, they perform only a limited amount of liquid holding recovery (LHR). Exposure of the dye with BrdUrd to near UV light in solution results in the appearance of two BrdUrd derived photoproducts. One appears to be deoxyuridine, and the other — an adduct of BrdUrd-dye. The adduct is acid labile and as a result only uracil is observed in acid-hydrolyzates of DNA after exposure of BrdUrd-substituted cells to 360 nm light in the presence of 33258 Hoechst. The production of uracil is linearly dependent on light exposure. Cells in which 85% of thymidine was replaced by BrdUrd are unable to remove more than 5–10% of uracil from their DNA during postirradiation incubation. However, when only 4% of thymidine is replaced, about 50% of the uracil is removed during 30min incubation after exposure. The results are consistent with our previous work, indicating that BrdUrd interferes with repair via excision-resynthesis. A working hypothesis is suggested to explain this interference.     

Photoreactivation in Paramecium tetraurelia under conditions of various degrees of ozone layer depletion

Photoreactivation (PR) is an efficient survival mechanism that helps protect cells against the harmful effects of solar-ultraviolet (UV) radiation. The PR mechanism involves photolyase, just one enzyme, and can repair DNA damage, such as cyclobutane-pyrimidine dimers (CPD) induced by near-UV/blue light, a component of sunlight. Although the balance of near-UV/blue light and far-UV light reaching the Earth's surface could be altered by the atmospheric ozone layer's depletion, experiments simulating this environmental change and its possible effects on life have not yet been performed. To quantify the strength of UVB in sunlight reaching the Earth's surface, we measured the number of CPD generated in plasmid DNA after UVB irradiation or exposure to sunlight. To simulate the increase of solar-UV radiation resulting from the ozone layer depletion, Paramecium tetraurelia was exposed to UVB and/or sunlight in clear summer weather. PR recovery after exposure to sunlight was complete at a low dose rate of 0.2 J/m2 x s, but was less efficient when the dose rate was increased by a factor of 2.5 to 0.5 J/m2 x s. It is suggested that solar-UV radiation would not influence the cell growth of P. tetraurelia for the reason of high PR activity even when the ozone concentration was decreased 30% from the present levels.     

THE EXPRESSION OF LIQUID HOLDING RECOVERY IN ULTRAVIOLET-IRRADIATED ESCHERICHIA COLI REQUIRES A DEFICIENCY IN GROWTH MEDIUM-DEPENDENT DNA REPAIR

Ultraviolet (UV) irradiated Escherichia coli K-12 recA cells (but not rec⁺ cells) show enhanced survival if they are held in buffer prior to plating for viability. To understand the role of the recA mutation in this liquid holding recovery (LHR) phenomenon, we have studied LHR in a temperature sensitive recA 200 mutant. The detection of LHR requires that the irradiated cells be recA when they are plated on growth medium, but the recA deficiency plays no role during liquid holding (LH). We conclude that it is the extreme sensitivity of recA cells in growth medium to unrepaired DNA daughter-strand gaps that magnifies the beneficial effects of the excision repair of DNA lesions during LH. Furthermore, we demonstrate a correlation between a strain's inability to perform growth medium dependent repair and its ability to express LHR. The relative amount of LHR was: recA > recF > lexA > recB > wild type (with the recB and wild-type strains showing negative LHR). Two strains did not show this correlation; the uvrD strain showed less LHR than expected from its UV radiation sensitivity, while the polA strain showed more. The molecular bases for these exceptions are explored.     

Differential Responses of Growth and Photochemical Performance of Marine Diatoms to Ocean Warming and High Light Irradiance

Diatoms have relatively high biomass in mid- to high-latitude oceans, which is also the most sensitive region to climate change. Photoautotrophs are thus predicted to become exposed to both higher temperatures and increased solar irradiance. In this study, we examined the consequences of such changes for the growth and photo-physiology of two diatoms by mimicking the scenarios that correspond to present day and that predicted for the end of this century. Elevated light induced higher rates of damage to photosystem II (PSII) that significantly reduced photochemical yields of both diatoms. Treatments including UV radiation induced ~ 50% inhibition of PSII under present PAR levels. Generally, warming alleviated UVR inhibition, resulting in higher photochemical yields, and faster recovery during dim light exposure. Therefore, concurrent increase of irradiance and temperature mitigated UV inhibition of PSII by 8–15%. The growth was stimulated by warming under PAR treatment, while less stimulation, or even decreased growth rates were found under the PAR + UVR treatment. Results suggest that ocean warming could fully offset the inhibition of high light on PSII. However, under the latter higher UVR stress scenario, the energetic expenditure required by the diatoms to repair damage could lead to their lower overall growth in future oceans.     

Synergistic Interaction of Photodynamic Treatment with the Sensitizer Aluminum Phthalocyanine and Hyperthermia on Loss of Clonogenicity of CHO Cells

Abstract— When CHO cells were exposed to hyperthermia and subsequently to photodynamic treatment, the combined effects were additive but in the reverse sequence the interaction was synergistic. The synergistic interaction comprised two quite different components: (1) photodynam-ically induced sensitization of cellular proteins and/or supramolecular structures for thermal inactivation and (2) a photodynamically induced inhibition of the cellular repair system for sublethal thermal damage. The first component of the synergistic interaction was reflected by a change of the Arrhenius parameters of thermal cell killing. A lowering of the activation energy of this process was responsible for the synergistic interactions, whereas a concomitant decrease of the frequency factor, opposing this effect, actually caused a much lower degree of synergism at higher temperatures. This component of the synergistic interaction did not respond to the insertion of an intermediate incubation period between the two treatments. The second component of the synergistic interaction, viz. the interference with the ability of cells to survive sublethal thermal damage, was reversible, as an intermediate incubation between photodynamic treatment and hyperthermia resulted in its repair. The photodynamically induced inhibition of the ability of cells to survive sublethal thermal damage was not related to ATP or glutathione depletion, inhibition of de novo protein synthesis or impairment of degradation of damaged protein molecules. Restoration of the repair system for sublethal damage depended on a metabolic process and required free intracellular Ca²⁺, suggesting that a cell signaling pathway may be involved. Thus, in a practical sense the magnitude of the synergistic interaction between photodynamic treatment and hyperthermia depends on the length of the interval between the two treatments and on the temperature and duration of the subsequent thermal treatment. This may have significant consequences for the development of clinical protocols.     

The Effects of UV Emission from Compact Fluorescent Light Exposure on Human Dermal Fibroblasts and Keratinocytes In Vitro

Compact fluorescent light (CFL) bulbs can provide the same amount of lumens as incandescent light bulbs, using one quarter of the energy. Recently, CFL exposure was found to exacerbate existing skin conditions; however, the effects of CFL exposure on healthy skin tissue have not been thoroughly investigated. In this study, we studied the effects of exposure to CFL illumination on healthy human skin tissue cells (fibroblasts and keratinocytes). Cells exposed to CFLs exhibited a decrease in the proliferation rate, a significant increase in the production of reactive oxygen species, and a decrease in their ability to contract collagen. Measurements of UV emissions from these bulbs found significant levels of UVC and UVA (mercury [Hg] emission lines), which appeared to originate from cracks in the phosphor coatings, present in all bulbs studied. The response of the cells to the CFLs was consistent with damage from UV radiation, which was further enhanced when low dosages of TiO₂ nanoparticles (NPs), normally used for UV absorption, were added prior to exposure. No effect on cells, with or without TiO₂ NPs, was observed when they were exposed to incandescent light of the same intensity.     

THE PHOTODYNAMIC EFFECTS ONV–79 CHINESE HAMSTER CELLS

Abstract— Holding of acriflavine sensitizedV–79 cells in growth medium before visible light exposure decreases inactivation by visible light. The decrease depended upon the period of holding, indicating that there was release of cellular dye during this period. Exposures to visible light were done in two conditions: (a) with no dye in the medium during visible light exposure (washed) and (b) with dye in the medium during exposure (unwashed). Caffeine was found to slightly increase the sensitivity of the cells to visible light in the washed condition, whereas, in the unwashed condition no such effect was observed. Interaction studies with far UV did not reveal any correlation between photodynamic damage and UV damage. Visible light exposure of acriflavine sensitized cells was found to be mutagenic, as studied from the induction of 8-azaguanine resistant mutants. Inhibition of singlet oxygen production by sodium azide suppressed the induction of mutants. All these, taken together, have been discussed with respect to the relative importance of DNA and non-DNA damage in the photodynamic action of acriflavine.     

Noninvasive Assessment of UV-induced Skin Damage: Comparison of Optical Measurements to Histology and MMP Expression

Acute exposure to UV radiation (UVR) causes visible skin damage such as erythema and results in local and systemic immunosuppression while chronic exposure can result in photocarcinogenesis. These deleterious effects can be quantified by histology and by bioassays of key biological markers, including matrix metalloproteinases (MMPs), or tryptophan moieties. We now report our results in quantifying UV skin damage with noninvasive optical methods based on reflectance and fluorescence spectroscopy and compare these noninvasive measurements to histopathology and MMP-13 expression. A solar simulator with spectral output nearly identical to that of solar radiation was developed and used in our experiments. SKH1 hairless mice were exposed to solar-simulated UVR at a total dose of 21 MED delivered over 10 weeks. Changes in oxygenated and deoxygenated hemoglobin were measured by diffuse reflectance spectroscopy, and tryptophan changes were monitored via a fluorescence monitor. Our results show that there is an increase in erythema, skin fluorescence, sunburn cells and MMP-13 after a series of suberythemal doses of UV irradiation on a hairless mouse animal model. Increased skin fluorescence is observed with increasing UV exposure. The levels of MMP-13 increase as the cumulative UV dose increases but their increase does not correspond to noninvasively measured changes.     

Kinetics of UV light-induced cyclobutane pyrimidine dimers in human skin in vivo: an immunohistochemical analysis of both epidermis and dermis

It is well known that UV exposure of human skin induces DNA damage, and the cumulative effect of such repeated damage is an important contributor to the development of skin cancer. Here, we demonstrate UV dose- and time-dependent induction of DNA damage in the form of cyclobutane pyrimidine dimers (CPD) in skin cells following a single exposure of human skin to UV radiation. CPD+ cells were identified by an immunohistochemical technique using monoclonal antibodies to thymine dimers. The percentage of CPD+ cells was UV dose-dependent, even a suberythemal (0.5 minimal erythemal dose [MED]) dose resulted in detectable level of cells that contained pyrimidine dimers. Forty-eight hours after irradiation the percent of total epidermal cells positive for CPD ranged from 19 +/- 8, 36 +/- 10, 57 +/- 12 and 80 +/- 10, and total percent dermal cells positive for CPD ranged from 1 +/- 1, 7 +/- 3, 16 +/- 3 and 20 +/- 5, respectively, following 0.5, 1.0, 2.0 and 4.0 MED. CPD were also observed in deeper reticular dermis, which suggest the penetrating ability of UV radiation into the skin. The change in CPD+ cells from 0.5 to 240 h post-UV exposure in both epidermal and dermal compartments of the skin was also quantitated. CPD+ cells were observed in skin biopsies at early time points after UV exposure which remained elevated for 48 h, then declined significantly by 3 days post-UV. A close examination of the skin at and after 3 days following UV exposure indicates the significant removal of DNA damaged cells from the epidermis. Ten days after UV exposure the levels of CPD+ cells in both epidermis and dermis were not significantly different from that in unirradiated skin.     

Accidental exposure to UV radiation produced by germicidal lamp: case report and risk assessment

Ultraviolet radiation is known to cause both benefits and harmful effects on humans. The adverse effects mainly involve two target organs, skin and eye, and can be further divided into short- and long-term effects. The present case report describes an accidental exposure of two health-care workers to ultraviolet radiation produced by a germicidal lamp in a hospital pharmacy. The germicidal lamp presented a spectrum with an intense UV-C component as well as a modest UV-B contribution. Overexposure to UV-C radiation was over 100 times as large as the ICNIRP exposure limits. A few hours after the exposure, the two subjects reported symptoms of acute UV injury and both of them continued having significant clinical signs for over 2 years. In this study, we describe acute and potentially irreversible effects caused by high UV exposure. In addition, we present the results of risk assessment by occupational exposure to germicidal lamps.     

Photopolymerization kinetics and phase behaviour of acrylate based polymer dispersed liquid crystals

Polymer dispersed liquid crystal (PDLC) materials were prepared by a polymerization induced phase separation (PIPS) process using UV radiation. The samples were obtained from the liquid crystalline component E7 and a blend of monofunctional (2-ethylhexyl acrylate) and difunctional (1,6-hexane diol diacrylate) monomers as precursors of the matrix. Polymerization energetics and thermodynamic properties of PDLC materials were studied by differential scanning calorimetry. Photopolymerization kinetics and phase behaviour of the PDLC are presented and discussed as a function of UV polymerization conditions (UV light intensity, UV time exposure and isothermal curing temperature).     

How does the skin sense sun light? An integrative view of light sensing molecules

The consensus on the effects of excessive sun exposure on human health has long emphasized the negative effects of solar UV radiation. Nevertheless, although UV radiation has been demonized, less is known about the consequences of sun exposure while using sunscreen, which can lead to high visible light exposure. UV and visible light play key roles in vitamin D synthesis, reduction of blood pressure, among other beneficial effects. In this review, we aim to provide a comprehensive view of the wide range of responses of the human skin to sunlight by revisiting data on the beneficial and harmful effects of UV and visible light. We start by exploring the interaction of photons in the skin at several levels including physical (depth of photon penetration), chemical (light absorption and subsequent photochemical events), and biological (how cells and tissues respond). Skin responses to sun exposure can only be comprehensively understood through a consideration of the light-absorbing molecules present in the skin, especially the light-sensing proteins called opsins. Indeed, many of the cellular responses to sun exposure are modulated by opsins, which act as the “eyes of the skin”.     

Rat lens glycolysis after in vivo exposure to narrow band UV or blue light radiation

UV radiation and short wavelength visible light are known to damage various tissues in the eye. This paper investigates the effect on rat lens glycolysis after in vivo exposure with 90 kJ m⁻² narrow band UV radiation (UVB, 300 nm) and 90 kJ m⁻² blue light (435 nm) radiation. After exposure, all lenses were incubated in Medium 199. Samples of culture medium were withdrawn after 2, 4, 6 h and 5, 10, 20 h in two UVB studies and after 5, 10 and 20 h in a blue light study. Lactate is the major end product of lens glycolysis. Lactate was determined with a modified enzymatic-photometric method. Intralenticular lactate was determined in one UVB experiment. In the UVB experiments we found a lower lactate production in the exposed lenses 2–6 h after exposure. There was an accumulation of lactate inside UVB-exposed lenses after 6 h incubation compared with their contralateral lenses. No significant effect on lactate production was observed in the blue light experiment. Conclusions. UVB induced a reversible inhibition of glycolysis. UVB also induced an accumulation of lactate inside the lens. Blue light tended to increase glycolysis.     

REPAIR OF UV DAMAGE IN ESCHERICHIA COLI UNDER NON-GROWTH CONDITIONS

Abstract. –A large difference in survival occurs between buffered suspensions of E. coli irradiated with UV radiation at a low fluence rate and those irradiated at a high fluence rate. For sufficiently large fluences, the extent of this fluence rate dependent recovery (FRR) is about two orders of magnitude greater than that which can be brought about by liquid holding recovery (LHR) following high fluence rate irradiation in most of the E. coli strains studied. LHR and FRR occur in excision resynthesis repair proficient (ERR⁺) but not ERR^- strains of E. coli , although its observation can be masked in strains with complete repair potential upon subsequent growth on nutrient plates. Accumulation of DNA strand interruptions and excision of cyclobutyl dipyrimidine occur during LHR and FRR but are more extensive for the latter. Our data suggest mat events beyond incision and excision occur during LHR and FRR, but differences in the extent of ERR during LHR and FRR cannot account for the difference in cell survival between these two phenomena.     

Development and characterization of a DNA solar dosimeter

In this paper, we report the development and characterization of a solar ultraviolet (UV) dosimetry system that can be used as a film badge for radiation monitoring. DNA molecules are coated on a thin nylon membrane as a UV dosimeter. The membrane is sealed in a polyethylene filter envelope with silica gel to keep the humidity low. After exposure to UV or solar light, induced DNA damage is measured by an immunochemical reaction. The intensity of color developed during the immunological reaction can be correlated linearly with the irradiated UV dose delivered by an Oriel solar simulator within a limited dose range. We observe no effects of temperature on the level of damage induction. The membrane is proficient for measuring DNA damage for more than 21 days when stored at either 37 or 4°C. The induced damage remains stable on the membrane for at least 22 days at both 37 and 4°C. In addition to these indoor experiments, we report measurements of solar UV dose in outdoor experiments.     

UV released factors induce antioxidant defense in A375 cells

UV light leads to release of different secretory factors from irradiated cells of which some of them have been characterized. We have reported earlier that cells exposed to the supernatant medium from irradiated cells were resistant to killing by some genotoxic agents. In this study, we present our finding that demonstrates DNA damage induced by UV or H(2)O(2) is lowered on prior exposure to the UV released factors (UVRF). Production of ROS in cells and lipid peroxidation was also lowered. It was found that treatment of unexposed cells with UVRF present in the supernatant medium altered the antioxidant defense activity in cells. Significant was the increase in catalase (CAT) and Cu-Zn superoxide dismutase (SOD) activity, whereas glutathione peroxidase (GPx) and reduced glutathione (GSH) levels remained unaffected. Cells exposed to UVRF prior to UV or H(2)O(2) treatment also experienced such upregulation; however, the remarkable increase in the GPx activity exhibited by these cells was not observed in cells exposed to H(2)O(2) or UV alone. It appears that exposure to UVRF tinkered with antioxidant defense in cells to facilitate its proliferation upon assault by an agent that can produce oxidative damage.     

REPAIR OF 313-NM INDUCED LESIONS AND PHOTOPROTECTION IN YEAST CANDIDA GUILLIERMONDII

Abstract. The present communication is concerned with the effects of near-UV radiation (300–380 nm) on yeast Candida guilliermondii. It was found that certain doses of 313 nm irradiation caused inactivation of the yeast which was exhibited in a way different from the lethal action of far-UV radiation. It was also found that the cells inactivated by 313 nm are capable of recovering vitality, if incubated for some time in a non-nutrient medium. The yeast inactivated by far-UV radiation also proved to be capable of recovering, though to a lesser degree. Both 334 nm radiation and non-lethal doses at 313 nm induced the photoprotective effect against far-UV damage. The effect was exhibited if there was a certain time interval (2–4 h) between the exposures to photoprotective light and subsequent far-UV radiation. Within this time interval the extent of photoprotection was dependent on temperature.     

PHOTOAGING EFFECTS ON SPECTRAL TRANSMITTANCE OF PLASTIC FILTERS

We examined the effects of ultraviolet (UV) radiation in combination with high levels of infrared (IR) radiation on the spectral transmittance of plastic filters. The biological action spectrum for damage to the human eye and skin changes dramatically in the 300-400 nm wavelength range. Cut-off filters used in this region to shape the spectrum of exposure sources are thus critical to the design of experiments which use broadband light sources. The changes in transmittance of three types of plastic filters were observed over an exposure period of 1000 h. One set of three filters was exposed mainly to UV radiation, while the other set was exposed to UV radiation plus IR radiation. Filters exposed to both UV and IR radiation showed spectral changes in their transmittance, while the filters exposed to UV only showed no measurable changes.