Effects of Light Environment During Culture on UV-lnduced Cyclobutyl Pyrimidine Dimers and Their Photorepair in Rice (Oryza sativa L.)

Abstract— We examined the effects of a light environment during culture of rice plants ( Oryza sativa ) on the steady-state cyclobutyl pyrimidine dimer (CPD) level, CPD induction by challenge UVB exposure and the ability to photorepair CPD. The steady-state CPD level in plants grown under visible radiation with supplemental UVB radiation in a growth chamber was several times higher than in plants grown without supplemental UVB radiation, whereas in outdoor-grown plants, it was not enhanced by supplemental UVB radiation. The susceptibility to CPD induction by challenge UVB exposure was highest in dark-grown plants and decreased with increasing irradiance of visible radiation at low and high levels and outdoors. Chronic UVB radiation reduced the susceptibility to UV-induced CPD in plants grown both indoors and outdoors. There was a significant negative correlation between CPD levels induced by challenge UVB exposure and the content of UV-absorbing compounds. The UV-induced CPD could be reduced by subsequent blue radiation in all samples except in dark-grown seedlings. The higher the irradiance of visible radiation in the culture, the greater the ability to photorepair CPD. Chronic UVB radiation did not increase the ability to photorepair CPD.     

Oxidation of guanine in cellular DNA by solar UV radiation: biological role.

The formation of cyclobutane pyrimidine dimers (CPD) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) was investigated in Chinese hamster ovary cells upon exposure to either UVC, UVB, UVA or simulated sunlight (SSL). Two cell lines were used, namely AT3-2 and UVL9, the latter being deficient in nucleotide excision repair and consequently UV sensitive. For all types of radiation, including UVA, CPD were found to be the predominant lesions quantitatively. At the biologically relevant doses used, UVC, UVB and SSL irradiation yielded 8-oxodGuo at a rather low level, whereas UVA radiation produced relatively higher amounts. The formation of CPD was 10(2) and 10(5) more effective upon UVC than UVB and UVA exposure. These yields of formation followed DNA absorption, even in the UVA range. The calculated relative spectral effectiveness in the production of the two lesions showed that efficient induction of 8-oxodGuo upon UVA irradiation was shifted toward longer wavelengths, in comparison with those for CPD formation, in agreement with a photosensitization mechanism. In addition, after exposure to SSL, about 19% and 20% of 8-oxodGuo were produced between 290-320 nm and 320-340 nm, respectively, whereas CPD were essentially (90%) induced in the UVB region. However, the ratio of CPD to 8-oxodGuo greatly differed from one source of light to the other: it was over 100 for UVB but only a few units for UVA source. The extent of 8-oxodGuo and CPD was also compared to the lethality for the different types of radiation. The involvement of 8-oxodGuo in cell killing by solar UV radiation was clearly ruled out. In addition, our previously reported mutation spectra demonstrated that the contribution of 8-oxodGuo in the overall solar UV mutagenic process is very minor.     

Development and application of a novel immunoassay for measuring oxidative DNA damage in the environment

We developed a facile, cost-effective competitive binding assay for the analysis of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) in DNA, using a polyclonal rabbit antiserum raised against an 8-oxodGuo hapten coupled to bovine serum albumin and radiolabeled synthetic ligand containing multiple 8-oxodGuo residues. This radioimmunoassay (RIA) displays a high affinity for 8-oxodGuo in DNA, with a detection limit of approximately 1 adduct in 10(5) bases of DNA. 8-oxodGuo standards for RIA were quantified by high-performance liquid chromatography and electrochemical detection in DNA diluted in methylene blue and exposed to visible light. As an initial application we quantified 8-oxodGuo in dosimeters deployed at increasing depths in the Southern Ocean during the austral spring of the 1998 field season or at the surface at Palmer Station, Antarctica, throughout the 1999 field season. Cyclobutane pyrimidine dimers (CPD) were quantified using an established RIA. We found that the frequency of both photoproducts decreased with depth. However, CPD induction was attenuated at a faster rate than 8-oxodGuo, correlating with the differential attenuation of solar ultraviolet wavelengths in the water column. CPD induction was closely related with ultraviolet-B radiation (UVB) attenuation, whereas the lower attenuation of 8-oxodGuo suggests that oxidative damage is more closely related to ultraviolet-A radiation (UVA) irradiance. The ratio of 8-oxodGuo: CPD was also found to covary with changes in stratospheric ozone concentrations at Palmer Station. These data demonstrate the usefulness of these assays for environmental photobiology and the potential for their use in studying the relative impacts of UVB versus UVA, including ozone depletion events.     

TNO Institute of Environmental Sciences, Energy Research and Process Innovation, the Netherlands

Abstract— A radiative transfer model is used to study the effect of stratospheric ozone depletion and tropospheric pollution on UVB (280-315 nm) radiation in the troposphere at midiatitudes of the Northern Hemisphere. The difference in the vertical distribution of UVB radiation for a range of pollution conditions typical for remote clean and more polluted sites on cloudless summer days is shown. The changes in downward UVB irradiance, UVB actinic flux, DNA-weighted UV radiation and photodissociation frequency of ozone at the earth's surface are quantified in relation to stratospheric ozone depletion and typical ranges of tropospheric pollutants (ozone [O3], sulfur dioxide [SO₂] and aerosols) in nonurban polluted regions. The results show that near the earth's surface, in non-urban polluted regions, an increase of tropospheric pollution, which occurred over the last 50-100 years, has cancelled the effect of an increase of UVB radiation due to stratospheric ozone depletion that has occurred since the late 1970s. The results hold for summer conditions and the decrease of UVB radiation is most pronounced in the polluted boundary layer. At higher altitudes the effect of tropospheric pollution on UVB radiation is far less. Prior to 1970 it was mainly tropospheric pollution that altered UVB radiation at the surface of the earth. In addition, since the late 1970s enhanced UVB radiation reaching the top of the troposphere, due to stratospheric ozone depletion, also had an impact on UVB levels in the troposphere.     

Diel Cycles of DNA Damage and Repair in Eggs and Larvae of Northern Anchovy, Engraulis mordax, Exposed to Solar Ultraviolet Radiation

Abstract— Damage from UVB radiation (280–320 nm) in the form of cyclobutane pyrimidine dimers (CPD) in DNA and the capacity for their repair were measured in newly spawned eggs and yolk-sac larvae of northern anchovy, Engraulis mordax, exposed to natural diel cycles of sunlight. The CPD were measured by a newly developed chemiluminescent immunoblot assay capable of measuring CPD in samples as small as 50 ng DNA. Eggs and yolk-sac larvae exposed to full irradiance levels died. At lower dose rates, equivalent to deeper more natural locations in the water column, there was a diel cycle of dimer concentration that tracked solar intensity. This diel cycle was due to the interaction of damage and repair processes. Repair of CPD in anchovy eggs and larvae could be attributed to true photodependent repair that could be stopped by moving samples into the dark. The CPD present at sunset remained until the following morning. The diel cycles of damage and repair were maintained over at least 4 days without a long-term upward or downward trend in dimer concentration. This indicates that at the UVB doses used for these experiments, there was no long-term accumulation of CPD nor an induction of increased repair capacity. Unhatched embryos spawned in the dark also exhibited a strong photorepair response, suggesting that photolyase expression was innate and not dependent on previous light exposure. The diel cycle observed here indicates that, at least for northern anchovy, the CPD concentration at the time of sampling is a good indicator of dose rate but a poor indicator of cumulative dose (i.e. late afternoon samples have the highest cumulative dose but relatively low CPD concentrations). The CPD immunoassay described here has the required sensitivity for measuring DNA damage in wild populations of ichthyoplankton exposed to natural sunlight. These results will guide the collection and interpretation of field data on natural levels of CPD in wild larvae collected at different depths and times of the day.     

Effects of cytosine methylation on pyrimidine dimer formation in DNA

The relative induction of cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4)pyrimidone photoproducts ([6-4]PD) was quantified in the duplex homopolymers polydeoxyadenosine:polydeoxythymidine, polydeoxyguanosine:polydeoxycytidine and polydeoxyguanosine:polydeoxy-5-methylcytidine irradiated with UVC or UVB radiation. Cytosine methylation significantly increased the yield of cytosine (6-4)PD after irradiation with UVC light and of cytosine CPD and (6-4)PD after irradiation with UVB light. The data suggest that CPD and (6-4)PD are preferentially induced at 5-methylcytosine bases in DNA of cells exposed to sunlight and comprise a major component of the mutation spectrum leading to the initiation of sunlight-induced skin cancer.     

Quantification of Cyclobutane Pyrimidine Dimers Induced by UVB Radiation in Conidia of the Fungi Aspergillus fumigatus,Aspergillus nidulans,Metarhizium acridum and Metarhizium robertsii

Conidia are responsible for reproduction, dispersal, environmental persistence and host infection of many fungal species. One of the main environmental factors that can kill and/or damage conidia is solar UV radiation. Cyclobutane pyrimidine dimers (CPD) are the major DNA photoproducts induced by UVB. We examined the conidial germination kinetics and the occurrence of CPD in DNA of conidia exposed to different doses of UVB radiation. Conidia of Aspergillus fumigatus, Aspergillus nidulans and Metarhizium acridum were exposed to UVB doses of 0.9, 1.8, 3.6 and 5.4 kJ m⁻². CPD were quantified using T4 endonuclease V and alkaline agarose gel electrophoresis. Most of the doses were sublethal for all three species. Exposures to UVB delayed conidial germination and the delays were directly related both to UVB doses and CPD frequencies. The frequencies of dimers also were linear and directly proportional to the UVB doses, but the CPD yields differed among species. We also evaluated the impact of conidial pigmentation on germination and CPD induction on Metarhizium robertsii. The frequency of dimers in an albino mutant was approximately 10 times higher than of its green wild-type parent strain after exposure to a sublethal dose (1.8 kJ m⁻²) of UVB radiation.     

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.     

UVB-lnduced DNA Damage and Its Photorepair in Nuclei and Chloroplasts of Spinacia oleracea L.

Ultraviolet-B-induced lesions and their photorepair in nuclear and chloroplast DNA of spinach (Spinacia oleracea L.) leaves were examined with two photoproducts, cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) pyrimidinone photoproducts (6-4PP). These photoproducts were induced both in nuclear and chloroplast DNA by UVB irradiation and could be detected by enzyme-linked immunosorbent assay using their respective monoclonal antibodies. Formation of CPD was greater in nuclear DNA than in chloroplast DNA (about 10 to 7), whereas 6-4PP formation was comparable in both DNA. On subsequent exposure of leaves to blue/UVA after UVB irradiation, photorepair of CPD and 6-4PP occurred in nuclear DNA but not in chloroplast DNA. When isolated chloroplasts were irradiated with UVB, CPD was also induced in their DNA. But photorepair of CPD did not occur in them by subsequent exposure to blue/UVA, suggesting that no photorepair system operates in chloroplasts.     

STRATOSPHERIC OZONE DEPLETION BETWEEN 1979 and 1992: IMPLICATIONS FOR BIOLOGICALLY ACTIVE ULTRAVIOLET-B RADIATION and NON-MELANOMA SKIN CANCER INCIDENCE

Abstract— The depletion of stratospheric ozone (0₃) has predictable implications for increases in biologically damaging solar ultraviolet-B radiation (UVB,280–320 nm) reaching the earth's surface. A radiative transfer analysis of satellite-based O₃ measurements between January 1979 and December 1992 shows that surface UVB levels increased substantially at all latitudes except the tropics, if other factors such as cloud cover and local pollutant levels have remained constant over this period. Exposure to UVB radiation is known to induce basal cell and squamous cell skin cancers, and dose-response relationships derived from epidemiological data can be combined with the UVB enhancements to estimate the seasonal and latitudinal distribution of future expected increases in the incidence of these cancers.     

Photoacclimation to long-term ultraviolet radiation exposure of natural sub-Antarctic phytoplankton communities: Fixed surface incubations versus mixed mesocosms

Solar UVB radiation (280-320 nm) is known to have detrimental effects on marine phytoplankton. Associated with the seasonal ozone hole in Antarctica, stratospheric ozone depletion occasionally influences the sub-Antarctic (Beagle Channel, Argentina) region, enhancing levels of UVB. The primary objective of this work was to study the effects of several (i.e. 6-10) days of exposure to UVB on the taxonomic composition and photosynthetic inhibition of local phytoplankton communities. For different light treatments, fixed-depth incubations placed in an outdoors water tank were compared with incubations in 1900 L mesocosms, where vertical mixing was present. Phytoplankton growth was inhibited by UV radiation (UVR) in fixed-depth experiments but not in the mixed mesocosms. Under fixed and mixed conditions alike, photosynthesis was significantly inhibited by UVB at the beginning of the experiment but no longer after several days of exposure, suggesting that cells had acclimated to radiation conditions. There was a change in species composition in response to UVR exposure in both experiments, which likely explained acclimation. In the community exposed to fixed conditions this change was from a phytoflagellate-dominated assemblage to a community with high relative abundance of diatoms after 6 days of exposure. UVA was responsible for most of the observed growth inhibition; however, the reduction in photosynthesis was produced by UVB. The reasons behind this variability in responses to UVR are associated with species-specific sensitivity and acclimation, and the previous light history of cells. In the community exposed in mesocosms, an assemblage codominated by phytoflagellates and diatoms was observed at the beginning of the experiments. After 10 days of exposure, green algae (Eutreptiella sp.) had increased, and phytoflagellates were the dominant group. The synthesis of mycosporine-like amino acids (MAAs), antioxidant enzymes and photosynthetic antenna pigments, in relation to repair and protection processes, may explain the reduced inhibition of both growth and photosynthesis that was observed in the phytoplankton community after several days of exposure. For environments such as the Beagle Channel seasonally exposed to the ozone hole, the results obtained from the fixed-depth experiments show that species can cope with UVR by means of MAA synthesis, while mixing would primarily promote a change in species composition and defense strategies.     

BIOLOGICALLY EFFECTIVE DOSES OF SUNLIGHT FOR IMMUNE SUPPRESSION AT VARIOUS LATITUDES AND THEIR RELATIONSHIP TO CHANGES IN STRATOSPHERIC OZONE

Using information on solar irradiance at different latitudes derived from a radiative transfer model and a detailed in vivo action spectrum for immune suppression in a murine system, we report here calculations of the "biologically effective" irradiance of sunlight for immune suppression. From 40 degrees N to 40 degrees S in summer, under normal stratospheric ozone concentrations this value ranged from 0.27 W/m2 (40 degrees N or S) to a peak of 0.33 W/m2 (20 degrees N or S) predicting that 50% immune suppression in the Balb/c mouse would occur after 21-26 min of sunlight exposure within this latitude range. We also found that the most effective wavelengths for immune suppression shift from a peak of 270 nm in the laboratory to near 315 nm in sunlight. Furthermore, using ozone depletion scenarios of 5 to 20%, at latitudes 20 degrees S and 40 degrees N, a 0.6% increase in biologically effective irradiance levels of solar UVB for immune suppression was predicted for each 1% decrease of ozone. This value rose to a nearly 1% increase for each 1% decrease in ozone at 60 degrees N latitude in wintertime. These data indicate that activation of immune suppression, in a murine model, requires relatively low levels of sunlight and that these levels are easily obtainable over most of the populated regions of the world. Since a UVB-activated photoreceptor, urocanic acid, regulates immune suppression in mice and since this same compound exists on other mammalian skin, including human skin, suppression of the mammalian immune system is predicted to increase if substantial stratospheric ozone depletion takes place.     

Evaluation of DNA dosimetry to assess ozone-mediated variability of biologically harmful radiation in Antarctica

In this study we investigated the use of a DNA dosimeter to accurately measure changes in ultraviolet B radiation (UVBR; 280-315 nm) under Antarctic ozone hole conditions. Naked DNA solution in quartz tubes was exposed to ambient solar radiation at Rothera Research Station, Antarctica, between October and December 1998 for 3 h during UVBR peak hours (1200-1500 h). Trends in UVBR-mediated DNA damage (formation of cyclobutane pyrimidine dimers [CPD]) were related to cloud cover, ozone-column depth and spectroradiometric measurements of ambient radiation. Ozone-column depths ranged from 130 to 375 DU during the study period, resulting in highly variable UVBR doses, from 1.6 to 137 kJ m(-2) over the 3 h exposure, as measured by spectroradiometry. There was a strong positive correlation (86%) between dosimeter CPD concentrations and DNA-weighted UVBR doses. Ozone depth was a strong predictor of DNA damage (63%), and there was no significant relationship between CPD formation and cloud cover. Subtle changes in spectral characteristics caused by ozone depletion were detected by the biodosimeter; the highest CPD concentrations were observed in October when ozone-mediated shifts favored shorter wavelengths of UVBR. We conclude that the DNA biodosimeter is an accurate indicator of biologically effective UVBR, even under highly variable ozone conditions.     

The induction of immunity to a protein antigen using an adjuvant is significantly compromised by ultraviolet A radiation

Ultraviolet (UV) radiation from sunlight causes skin cancer and inhibits priming of the immune system during vaccination. However the dose related effects of the different components of sunlight (UVA and UVB) are complex and require further investigation. Using ovalbumin as a model protein vaccine with saponin as adjuvant we show that both UVA and UVB can suppress the DTH response to a poorly immunogenic protein. Increasing doses of UVB induced increased levels of immunosuppression and tolerance. UVA however, caused a bi-phasic dose response with intermediate but not low or high doses causing primary immunosuppression. No dose of UVA caused significant tolerance. Similar results were observed in both C57BL/6 and Balb/c mice. Our data confirms the complex immunomodulatory dose effects of UVA and UVB for a protein antigen, and shows that both UVB and UVA can suppress immunity induced by a protein with adjuvant. This highlights the importance of considering sun exposure patterns in the future success of both preventing skin cancer development and enhancing vaccination regimes.     

Exposure to increased ambient ultraviolet B radiation has negative effects on growth, condition and immune function of juvenile Atlantic salmon (Salmo salar)

Atlantic salmon (Salmo salar) parr were exposed in two outdoor experiments, ranging in duration from 52 to 137 days, to spectral treatments: (1) natural sunlight (=present ambient UVB level), (2) solar radiation supplemented with enhanced UVB radiation from lamps simulating 20% or 8% stratospheric ozone loss or (3) UVB-depleted sunlight achieved by screening with Mylar-D film. The growth, condition and immune function of the salmon were quantified after treatments. Exposure to enhanced UVB radiation retarded growth, and decreased hematocrit value and plasma protein concentration. Further, enhanced UVB radiation affected plasma immunoglobulin concentration. The results demonstrate that juvenile Atlantic salmon are not able to fully adapt to increased ambient UVB levels in long-term exposures, and the interference with immune system function suggests a negative effect of UVB on disease resistance in Atlantic salmon.     

DNA Damage Induced by Late Spring Sunlight in Antarctica

Sunlight ultraviolet (UV) radiation constitutes an important environmental genotoxic agent that organisms are exposed to, as it can damage DNA directly, generating pyrimidine dimers, and indirectly, generating oxidized bases and single-strand breaks (SSBs). These lesions can lead to mutations, triggering skin and eye disorders, including carcinogenesis and photoaging. Stratospheric ozone layer depletion, particularly in the Antarctic continent, predicts an uncertain scenario of UV incidence on the Earth in the next decades. This research evaluates the DNA damage caused by environmental exposure to late spring sunlight in the Antarctic Peninsula, where the ozone layer hole is more pronounced. These experiments were performed at the Brazilian Comandante Ferraz Antarctic Station, at King’s George Island, South Shetlands Islands. For comparison, tropical regions were also analyzed. Samples of plasmid DNA were exposed to sunlight. Cyclobutane pyrimidine dimers (CPDs), oxidized base damage and SSBs were detected using specific enzymes. In addition, an immunological approach was used to detect CPDs. The results reveal high levels of DNA damage induced by exposure under the Antarctic sunlight, inversely correlated with ozone layer thickness, confirming the high impact of ozone layer depletion on the DNA damaging action of sunlight in Antarctica.     

Evaluation of photolyase (Photosome) repair activity in human keratinocytes after a single dose of ultraviolet B irradiation using the comet assay

Photosome is constituted of photolyases included in liposomes. Photolyase is a bacterial enzyme that can repair ultraviolet B (UVB)-induced cyclobutane pyrimidine dimers (CPD) in eukaryotic cells. A modified version of the alkaline comet assay has been set up to evaluate the repair activity of this enzyme after a single dose of UVB (312 nm, 0.06 J/cm2) in human keratinocytes. The formation of single strand breaks (SSB) induced by the UVA photoactivation of the enzyme (1.2 J/cm2) was inhibited by the pretreatment of the cells with 4 mM L-ergothioneine (ERT) during 30 min at 37 degrees C. To increase the sensitivity of the comet assay, an additional lysis was used with a buffer containing sodium dodecyl sulfate (0.5%) and proteinase K (0.1 mg/ml) for 60 min at 37 degrees C. Unrepaired CPD by photolyase were revealed by a second enzymatic treatment with T4 endonuclease V, a CPD specific glycosylase. UVB irradiation increased the SSB level in keratinocytes and additional T4NV treatment enhanced this SSB level by 1.5-2.0-fold confirming that CPD were the major base modifications generated by UVB irradiation. UVA-photoactivated Photosome repaired CPD lesions and decreased the SSB levels by 2.6-3.3-fold. Photosome could be an additional component of sunscreens to reduce the development of skin cancer.     

The Influence of Photosynthetically Active Radiation on the Effects of Ultraviolet-B Radiation on Arabidopsis thaliana

Ultraviolet-B (UVB;280–320 nm) radiation is a small but biologically significant portion of the solar spectrum reaching the earth's surface. Research interests have been fostered because UVB has been increasing in recent years due to depletion of stratospheric ozone. Ultraviolet-B that penetrates into plant tissue may damage important cellular macromolecules. Although there has been considerable research on the effects of UVB on plants, the influence of the level of photosynthetically active radiation (PAR;400–700 nm) on effects of UVB requires further definition as a prelude to studies of UVB sensitivity and defense mechanisms. Arabidopsis thaliana wildtype ecotype Landsberg erecta (LER), which is relatively insensitive to UVB, and the relatively sensitive LER-based mutant transparent testa-5 (tt5), were grown under 100 or 250 μmol m^?2 s^?1 PAR and then exposed to O or 7 kJ m^?2 day ^?1 UVB_BE under these PAR levels. Plants exposed to UVB had reduced dry weight and leaf area and higher levels of UV-absorbing compounds in leaf tissue. The level of PAR did influence the effects of UVB, with the higher level of PAR prior to UVB exposure reducing sensitivity of LER to UVB. In contrast to other studies, higher PAR supplied simultaneously with UVB increased rather than decreased sensitivity of both genotypes to UVB. These results demonstrate the importance of controlling and comparing PAR levels when undertaking studies of UVB sensitivity, as effects of UVB on plants are influenced by the PAR levels plants are growing under prior to and during exposure to UVB.     

UVB-induced Cyclobutane Pyrimidine Dimer Frequency Correlates with Skin Cancer Mutational Hotspots in p53

Abstract— Ultraviolet light has been identified as the major carcinogen in skin cancer and the p53 tumor suppressor gene is a major target for UV-induced mutations. The mutations are probably caused by unrepaired UV-induced cyclobutane pyrimidine dimers (CPD) and possibly by the less frequent pyrimidine (6-4) pyrimidone photoproducts. While hot spots for p53 mutations in human nonmela-noma skin tumors correspond quite well to slow spots for CPD repair in cultured cells irradiated with the model mutagen 254 nm UVC (which is not present in terrestrial sunlight), they do not all coincide with sequences that are initially frequently damaged by 254 nm UVC. Using LMPCR (ligation-mediated polymerase chain reaction), we show that environmentally relevant UVB light induces CPD at CC and PyrmC positions much more frequently than does UVC light, and that all eight skin cancer hot spots in p53 are also hot spots for UVB-induced CPD. Our results show that methylation of dipyrimidine sites (PyrmCpG) is associated with an increase rate of CPD formation upon UVB irradiation. Consequently, DNA methylation may increase the mutagenic potential of UVB and explains that several p53 mutation hot spots are found at PyrmCpG. The distribution patterns of CPD formation and the photofootprint patterns found along exons 5 and 6 of p53 gene are suggestive of DNA folding into nucleosomes.     

SEASONAL PHOTOSENSITIVITY OF BLEPHARISMA: THE ROLE OF TEMPERATURE

Red blepharismas (B. japonicum) in Pyrex test tubes exposed near noon to clear summer daylight are killed. If similarly exposed at dawn, they turn blue as the red pigment blepharismin is oxidized to oxyblepharismin. and they resist further exposure to daylight. Red blepharismas similarly exposed to sunlight at dawn in winter are killed. The experiments reported seek the reasons for the seasonal difference in photosensitivity and the way in which blepherismas survive winter. Red blepharismas (B. japonicum) exposed to visible and near ultraviolet (UV) radiation from two 4W coolwhite fluorescent lamps (5 Wm^-2; 2151ux) resist an 8h exposure between 15° and 34°C. turning blue. Blue and colorless blepharismas also resist 8h exposures to the light at 15° and 34°C. At 8°C both red and blue populations are killed, death appearing to result from deranged water regulation; colorless ones are not affected. Red blepharismas are killed by fluorescent light at 8° C even when the near-UV radiation is largely excluded by a Corning glass filter; blue and colorless ones are not. The reds are also killed at all temperatures by sunlight filtered through Pyrex glass if exposed near noon on a sunny day. At temperatures above 8° C they are not killed by direct sunlight passed through glass if exposed at dawn, but they turn blue. Blepharismas in fluorescent light divide at the same rate as controls in the dark, but they get smaller. When exposed to sunlight at dawn, at temperatures above 8°C. blepharismas in Pyrex test tubes containing stones or fruiting spikes of grass, hid in the shade of these objects, apparently by random movements. If exposed under similar conditions near noontime, the majority failed to find cover and were killed. Blepharismas survive in summer by avoiding sunlight, in winter by encystment. Colorless protozoans Paraineciurn multimicronucleatum, Colpidium colpoda and Spirostomum teres resisted killing in summer sunlight and with fluorescent light at all temperatures tested.