INDUCTION OF SKIN TUMORS IN HAIRLESS MICE BY A SINGLE EXPOSURE TO UV RADIATION*

Abstract— Skin tumors were induced in hairless mutant mice following a single exposure to ultraviolet radiation (UV). Tumors were first noted as early as 7 weeks following irradiation. The UV, emitted by FS20/40T12 fluorescent lamps, was principally in the 280–320 nm spectral region with a peak at 313 nm. Single (skin surface) doses of 3 times 10⁴ J/m² to 24 times 10⁴ J/m² were delivered in 3 h or less. The higher doses resulted in more severe acute damage as well as greater tumor yield. Most of the tumors were benign hyperplastic epithelial papillomas; 4 out of 96 tumors examined histologically proved to be squamous cell carcinomas. This appears to be the first report of experimental carcinogenesis due to a single UV exposure, not requiring exogenous chemical promotion.     

SUPPRESSION OF THE INDUCTION OF DELAYED-TYPE HYPERSENSITIVITY REACTIONS IN MICE BY A SINGLE EXPOSURE TO ULTRAVIOLET RADIATION

Abstract— After a single exposure of mice to UV radiation, their ability to generate a contact hypersensitivity (CHS) response to contact sensitizers applied epicutaneously to distant, unirradiated skin is severely impaired. It is not clear, however, if the classic delayed type hypersensitivity (DTH) reponse to exogenous antigens, injected into the subcutaneous (s.c.) space, can also be modulated by UV radiation. We report here that a single exposure of mice to UV radiation suppressed the induction of DTH to both erythrocyte and soluble protein antigens injected s.c., but did not suppress the elicitation of the response. The suppressive effect was abrogated by cyclophosphamide treatment. In addition, antigen-specific suppressor cells were found in the spleens of the mice with a decreased DTH response. Since the ability to mount a DTH response has been linked with the resistance to certain pathogenic microorganisms, we suggest that the suppression of DTH by UV radiation may have the potential to compromise host resistance to such infectious agents.     

INHIBITION OF THE IMMUNE RESPONSE TO ALLOANTIGEN IN THE RAT BY EXPOSURE TO ULTRAVIOLET RADIATION

Exposure of mice to ultraviolet radiation (UV) followed by alloantigen sensitization can suppress the immune response to that alloantigen. In order to assess the applicability of using UV-induced immunosuppression in organ transplantation, the effectiveness of UV in prolonging the survival of vascularized organ allografts must be determined. Because, for technical reasons, rats are better suited than mice for such experiments, we first wanted to determine whether UV suppresses the immune response of inbred rats to alloantigens. The data presented here demonstrate that exposure of rats to UV (115-129 kJ/m2) prior to alloantigenic sensitization decreases the mixed lymphocyte response to alloantigen. The depression of the proliferative response to alloantigen was selective in that spleen cells from the UV-treated rats could respond to mitogenic stimulation. In contrast to previous results with mice, suppressor cells could not be demonstrated in the spleens of the UV-treated rats. In addition, UV treatment after sensitization inhibited the response to alloantigen. These data suggest that treatment of the recipient with UV before or after alloantigenic sensitization may provide a novel method of inhibiting immune responses to allogeneic antigens.     

SKIN DAMAGE IN ADULT AMPHIBIANS AFTER CHRONIC EXPOSURE TO ULTRAVIOLET RADIATION

Abstract— The effects of repeated UV exposure on the skin of the European crested newt, Triturus cristatus carnifex , have been investigated. The animals were irradiated 3 times per week with a Westing-house FS40T12 fluorescent sun lamp (wavelength spectrum 275–350 nm). Two groups of animals received the same total fluence of 1.3 × 10⁵ J/m² in single fluences of either 1570 J/m² (group A) or 9430 J/m² (group C), and one group received a total fluence of 2.6 × 10⁵ J/m² in single fluences of 4710 J/m² (group B). All the animals were killed 7 months after the first UV exposure, but at different intervals after the last exposure. Striking epidermal hyperplasia was found in the newts irradiated at the lower fluence rate (group A). In the animals given the higher total fluence (group B), the most prominent skin changes were dermal fibrosis and irregular thinning and thickening of the epidermis. No significant skin changes were found in group C., in which if there had been UV lesions, they had been repaired during the 5 month interval between the last irradiation and the killing of the animals. No skin tumors developed in any experimental group.     

THE VASCULAR RESPONSE OF HUMAN SKIN TO ULTRAVIOLET RADIATION

The vascular response of human skin to 300 nm (UV-B) and 254 nm (UV-C) ultraviolet radiation was assessed using the reflectance measurement of erythema and the technique of laser Doppler velocimetry. For both wavelengths, the increase in measured Doppler blood flux varied with the increase in erythema in a quadratic manner predicted by a simple model based on the principles of fluid mechanics. This suggests that the mean red blood cell velocity increases significantly in areas of UV-B and UV-C erythema. No qualitative difference in response to these two wavelengths was demonstrated, suggesting that the same blood vessels are involved in the causation of both UV-B and UV-C erythema.     

ENHANCEMENT OF INTRINSIC ANTITUMOR ACTIVITY IN SPORE-ENDOTOXIN MIXTURES OF Bacillus thuringiensis BY EXPOSURE TO ULTRAVIOLET RADIATION

Abstract— Irradiation of spore-endotoxin mixtures from Bacillus thuringiensis cultures at 254 nm (60 μW cm^-2) enhances their intrinsic antitumor potency as well as that of either component. The extent of enhancement depends on the length of exposure (optimum: 35 min) and may thus be due to photochemical changes of the endotoxin protein or/and to photoproduction of additional compounds with antitumor activity. Antitumor effects, expressed as survival rates of C57BL/6 mice inoculated with Lewis' mouse lung carcinoma and subjected to treatments 24 h later, depended on the number of doses of preparations administered (mixture, separated components).     

FORMATION OF THYMINE DIMERS IN MAMMALIAN SKIN BY ULTRAVIOLET RADIATION IN VIVO

Abstract— Ultraviolet radiation of 220–300 nm is known to produce cyclobutyl pyrimidine dimers in extracellular DNA, in bacteria, and in mammalian cells in culture. The formation in vivo of such dimers in mammalian skin has remained inferential. We report that one of the important and recognizable biologic events that occurs in mammalian skin during irradiation is the formation of thymine dimers. [³H]-labelled thymidine was applied to the epilated skin of guinea pigs to label their DNA. Animals were irradiated individually, using wavelengths of either 254, 285–350, or 320–400 nm. Immediately after irradiation, epidermis was separated from the rest of the skin and homogenized; DNA and RNA were isolated. Irradiation with wavelengths of 285–350 nm, which included the sunburn-producing spectrum (i.e., 290–320 nm), produced thymine dimers (1·7–2·6 per cent of the total [³H]-thymine incorporated into DNA). Irradiation with 254nm also produced fewer dimers (0·46–1·2 percent); and 320–400 nm produced none. The dimer could be cleaved by 250 nm radiation to form thymine. The epidermal cell damage by ultraviolet radiation, particularly by the sunburn-producing spectrum (290–320 nm), may be related to the formation of such dimers.     

A DOSIMETRIC TECHNIQUE FOR THE MEASUREMENT OF ULTRAVIOLET RADIATION EXPOSURE TO PLANTS

Abstract Due to the effects of geometry, orientation, atmospheric conditions and optical properties of leaves and soils, the UV exposure to a plant may differ significantly from that of ambient radiation. This paper presents a method utilizing a passive measuring technique with polysulfone dosimeters that provides the ability of measuring the UV exposure at a number of sites simultaneously. The UV exposures are measured at selected sites over models of three canopy shapes with a computer program interpolating and summing these to provide the total UV exposure incident on the canopy.     

EVALUATION OF SKIN CANCER RISK RESULTING FROM LONG TERM OCCUPATIONAL EXPOSURE TO RADIATION FROM ULTRAVIOLET LASERS IN THE RANGE FROM 190 TO 400 nm

The relative risk of occupational exposure to radiation from UV lasers was estimated using a mathematical model based on both epidemiological data and animal experiments. Calculations were performed for the 193 nm ArF excimer laser cornea shaping, the 308 nm XeCl excimer laser for coronary angioplasty, and other UV lasers in a laboratory environment. The model included the effects of direct exposure and exposure to scattered radiation. The results show that for the two medical applications the increase in the relative risk is comparable to that of one additional day of sunbathing per year. For subjects exposed to UV lasers in a laboratory setting, the relative risk may increase to a value comparable to that of people with an outdoor profession.     

INDUCTION OF PHOTOPRODUCTS IN SYNTHETIC POLYNUCLEOTIDES BY FAR AND NEAR ULTRAVIOLET RADIATION

Single and double-stranded polynucleotides of thymine and cytosine have been used to analyse the photoproducts produced by irradiation with far or near UV light. Reversed-phase high performance liquid chromatography was used to detect and quantify cyclobutane dimers and Pyr(6–4)Pyo adducts produced by 254 nm. At 320 nm 10-times less Thy(5–6)Thy dimers and one half the number of Cyt(5–6)Cyt dimers were induced; no Pyr(6–4)Pyo adducts were evident. HPLC has recently been applied to the isolation and characterization of various nucleic acid substituents and their UV-photoproducts. The relative retention times of pyrimidines and their UV photoproducts on HPLC reflect differences in the hydrophobicity of the compounds being separated. Hence, the more hydrophobic (less polar) a compound is, the greater its capacity to bind to the column and the greater its retention time; for T o T, C<>T and C<>C dimers this difference may result from variations in the number of methyl moieties (Cadet et al., 1983). The retention time of the compound also depends on its stereochemistry. Separation of the four stereoisomers of T<>T by HPLC shows that compounds which are molecular equivalents can be more or less accessible to the non-polar stationary phase depending on their conformation (Cadet, 1980). The relatively long retention times of the bipyrimidine photoadducts suggest a structural configuration which allows greater access to the hydrophobic moieties of the molecule. It is intriguing to consider that this difference in conformation may also be reflected in DNA–protein interactions such as binding by UV-endonucleases or antibodies directed against UV photodamage. Proteins can be used as sensitive probes for photoproduct induction and repair but an accurate evaluation of their specificity is required. It was the intent of this paper not only to compare the induction parameters for various dimers in pyrimidine homo-polymers but to provide controlled substrates which can be used to define the relative binding efficiencies of repair enzymes and antibodies for different types of photoproducts.     

THE OCULAR DOSE OF ULTRAVIOLET RADIATION FROM SUNLIGHT EXPOSURE

Abstract— The ocular toxicity of ultraviolet radiation has been demonstrated in acute photokeratitis and is suspected of contributing to cataractogenesis and senile macular degeneration. While previous studies have emphasized photochemical and epidemiologic aspects of ocular UV-B irradiation, little is known about the extent of such exposure in human subjects. To determine levels of ocular UV-B exposure from sunlight, four mannikin headforms were fitted with UV-B sensitive film (polysulphone) and exposed on an unobstructed rooftop (Baltimore, Md.: latitude = 39.5 degrees) to four hours of sunlight (11 am-3 pm local time) over a three month period (June-August). Simultaneous measurements of ocular and ambient exposure revealed a ratio of 19.5 ± 2.9% that was independent of ambient level (P < 0.05). Measurements performed during earlier hours (8 am-11 am) revealed a similar ratio. Mannikin headforms fitted with brimmed baseball caps showed a22–95% reduction in ocular exposure, depending on the angle of the hat brim to the forehead. Three sets of spectacles substantially reduced ocular UV-B exposure,62–94% dependent on the absorption properties of the spectacle lenses. These anthropomorphic measurements indicate that a substantial percentage of ambient UV-B light is incident upon the cornea and that personal factors, such as wearing a hat or spectacles, can markedly affect UV-B exposure.     

EXPOSURE TO LONG WAVELENGTH ULTRAVIOLET RADIATION DECREASES PROCESSING OF LOW DENSITY LIPOPROTEIN BY CULTURED HUMAN FIBROBLASTS

Exposure of MRC5 human fibroblasts to UVA radiation (365 nm) resulted in a dose-dependent decrease in low density lipoprotein (LDL) uptake and degradation by cells. Following a 25 J/cm² irradiation dose, about 45% and 70% reduction in ¹²⁵I-LDL uptake and degradation were observed, respectively. Under the same conditions, the ¹⁴C-sucrose uptake was also decreased to about the same extent as LDL uptake. Cell pretreatment with the antioxidants vitamin E and vitamin C did not prevent the UVA-induced fall in LDL degradation. These results point to the possible effects of UVA radiation on receptor-mediated and nonspecific uptake of exogenous molecules. With special regard to the alterations in receptor-mediated processing of exogenous ligands, such a phenomenon could be of importance in UVA-induced skin degenerative processes.     

SKIN CANCER AND ULTRAVIOLET RADIATION

Abstract. A new model is presented for the calculation of the increased incidence of non-melanoma skin cancer in Caucasians resulting from ozone reduction. The model postulates that the probability of first incidence of such skin cancer is distributed log-normally as a function of total accumulated lifetime dose of harmful ultraviolet radiation. The effect on skin cancer incidence of an increase in harmful ultraviolet radiation due to ozone reduction can then be calculated directly from the extent to which each individual's lifetime accumulated dose is thereby increased. The result of such a perturbation, on average, would be to cause skin cancer to appear at a slightly earlier age. Since skin cancer is predominantly a disease of the elderly, this shift to younger ages has the effect, when integrated over the entire population, of increasing the overall total incidence of skin cancer.     

EFFECT OF IONIZING RADIATION UPON TOTAL DIFFUSE SPECTRAL REFLECTANCE OF ULTRAVIOLET LIGHT BY SKIN

Abstract— Irradiation of guinea-pig skin with X rays and beta particles resulted in decreased total diffuse reflectance (DSR) of 330–400 nm light. Qualitatively, this response resembled that seen after irradiation of the skin of normal guinea-pigs with ultraviolet (UV) radiation of wavelength shorter than 300 nm or that of photosensitized guinea pigs with UV wavelengths longer than 300 nm. We postulate that the transformations which depress the DSR result from energy-transfer processes, independent of the class of radiation. Moreover, they are intimately related to subsequent changes in vascular permeabilities (delayed erythema) which occur after the same radiation exposures that lower the 330–400 nm DSR of skin surfaces.     

ACTION SPECTRUM STUDIES FOR INDUCTION OF IMMUNOLOGIC UNRESPONSIVENESS TO DINITROFLUOROBENZENE FOLLOWING IN VIVO LOW DOSE ULTRAVIOLET RADIATION

Abstract— Topical application of the contact sensitizer dinitrofluorobenzene to the skin of C3H mice previously exposed in vivo to low doses of UVB radiation from FS20 sun lamps resulted in the acquisition of antigen-specific unresponsiveness to that hapten. When narrow band radiation at various wavelengths between 260 and 320 nm was employed, increasing doses of radiation were found to produce increasing amounts of immunosuppression. Construction of an action spectrum curve revealed that radiation between 260 and 300 nm was most efficient in producing unresponsiveness. Wavelengths above 300 nm were much less efficient than those below 300 nm. These results indicate that there are major differences in the effectiveness of various components of the short and middle wavelength UV spectrum in eliciting immunologic unresponsiveness to a topically administered hapten. Potential chromophores for this UVB-induced immunosuppressive effect include DNA and urocanic acid.     

PHOTOREACTIVATION OF ICR 2A FROG CELLS AFTER EXPOSURE TO MONOCHROMATIC ULTRAVIOLET RADIATION IN THE 252–313 nm RANGE

Abstract— Exposure of ICR 2A frog cells to photoreactivating light after treatment with monochromatic ultraviolet (UV) radiation in the 252–313 nm range resulted in an increase in survival with similar photoreactivable sectors for each of the wavelengths tested. As photoreactivating enzyme is specific for the repair of pyrimidine dimers in DNA, these findings support the hypothesis that these are critical lesions responsible for killing of cells exposed to UV radiation in this wavelength range. The action spectra for cell killing and production of UV-endonuclease sensitive sites were similar to the DNA absorption spectrum though not identical. Because the number of endonuclease sensitive sites is a reflection of the yield of pyrimidine dimers, these data also suggest that the induction of dimers in DNA by UV radiation in the 252–313 nm range is the principal event leading to cell death.     

SEQUENCE-SPECIFICITY OF THE ALKALI-SENSITIVE LESIONS INDUCED IN DNA BY HIGH-INTENSITY ULTRAVIOLET LASER RADIATION

The action of high-intensity (10(9)-10(12) W/m2) UV (266 nm) laser radiation pulses (duration ca 10 ns or ca 40 ps) on liquid aqueous solutions of DNA is known to cause not only single- but also two-quantum modification of nucleic bases. The action of hot piperidine on the laser-irradiated DNA results in non-random splitting of polynucleotide chain. Hence, at least some of the modified nucleoside residues are alkali-sensitive lesions (ASLs). The distribution of ASLs along the DNA chain shows that the position of these lesions corresponds with pyrimidines in the PyPy sequences (similar to those formed via single-quantum conversions) as well as with deoxyguanosine residues. The last ASLs result from two-quantum reactions and occur much more efficiently than the direct photo-induced cleavage of the internucleotide (phosphodiester) bond. It has been shown with fragments of plasmids pUC18, pUC19 and pBR322 (total length over 600 base pairs) that the relative efficiency of ASLs at deoxyguanosine sites depends on the primary structure context and can differ by an order of magnitude. The highest efficiency of modification is observed when a purine is 3' neighbour to the 2'-deoxyguanosine, i.e. at 5'-GPu-3' sites. However, considerable variations in the modification efficiency were also found in these sequences.     

THE DOSE-RESPONSE RELATIONSHIP OF TUMORIGENESIS BY ULTRAVIOLET RADIATION OF 254 nm

Abstract— Groups of albino hairless mice, Skh-hrl, were exposed daily to UVC radiation from low pressure Hg arcs (Philips TUV 40W). These lamps emit predominantly radiation of 254 nm. Three groups of animals were used in the experiments, each receiving a different daily dose.
The results were described with the Weibull probability function. As in earlier studies with UVB. the tumor induction time was proportional to a power of the daily dose. The exponent turned out to be as low as -0.2. This implies that the induction time varied only a little with the daily dose. The average number of tumors per animal was proportional to a power of time. A sample of 73 tumors of at least 4 mm in diameter were investigated histologically. The large majority were classified as squamous cell carcinomas.
A comparison was made with the results of an earlier reported experiment with Westinghouse FS40 sunlamps. Throughout the whole range of daily doses used in the present experiment, UVC was less carcinogenic than UVB. An intriguing difference between the two types of radiation was that the tumors induced by UVC appeared much more scattered over the irradiated parts of the animals than the UVB-induced tumors.     

DNA NICKING BY ULTRAVIOLET RADIATION IS ENHANCED IN THE PRESENCE OF IRON and OF OXYGEN

Double-stranded covalently closed circular supercoiled DNA (ccc DNA) from plasmid pUK 9 was irradiated in vitro at denned wavelengths in the UV region (290, 313 and 365 nm). The nicking was monitored by electrophoresis on agarose gels, ethidium staining and densitometric quantitation of supercoiled and relaxed moieties. At the explored wavelengths, the dose required for introducing one nick per million phosphodiester bonds diminishes with increased concentration of added ferric iron, whereas the effect of cupric iron is practically negligible. Adding metal chelators or bubbling argon prior to the irradiation results in a dramatic increase in the dose required for introducing one nick per million phosphodiester bonds. Taken together, these results seem to indicate that iron and oxygen play a role as cofactors in the UV-induced nicking of ccc DNA in vitro.