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Sunscreens are employed to mitigate the adverse effects of sunlight on skin but are primarily designed to prevent ultraviolet-B-associated burning and damage. The increasingly recognized role of ultraviolet A in aging, and possibly melanoma, highlights the need to include ultraviolet A screens; however, validation remains difficult. We have used a novel method to establish the efficacy of sunscreens, by measuring ultraviolet-A-induced free-radical production (thought to contribute towards ultraviolet-A-related aging and malignant change). Electron spin resonance spectroscopy was used to detect free radicals directly in human Caucasian skin during irradiation with levels of ultraviolet comparable to solar intensities. Using this system the protection afforded by three high factor sunscreens (sun protection factor 20+) that claim ultraviolet A protection was examined. Each sunscreen behaved similarly: at recommended application levels (≥2 mg per cm 2 ) the ultraviolet-induced free radicals were reduced by only about 55%, and by about 45% at 0.5–1.5 mg per cm 2 (0.5 mg per cm 2 reported for common usage). A "free-radical protection factor" calculated on the basis of these results was only 2 at the recommended application level, which contrasts strongly with the erythema-based sun protection factors (mainly indicative of ultraviolet B protection) quoted by the manufacturers (20+). The disparity between these protection factors suggests that prolonged sunbathing (encouraged by use of these creams) would disproportionately increase exposure to ultraviolet A and consequently the risk of ultraviolet-A-related skin damage.

Objectives: Both ultraviolet-A (UVA) and ultraviolet-B (UVB) radiation have been implicated in skin photoageing, a multifaceted phenomenon of clinical, cellular, histological, and immunological changes. Much of the in vivo evidence of the relationship between wavelength-dependent ultraviolet radiation exposure and photoageing has been based on animal models. This study considered the relative contributions of UVA and UVB radiation exposure to the hallmark feature of photoageing, degree of dermal elastosis, in an unselected human population. Methods: An age-strati®ed random sample of 338 adults aged over 17 years was chosen from the electoral roll of the community of Nambour, Australia. Subjects were recruited between May 1994 and May 1995, and asked to provide a 2-mm punch biopsy of the skin of the back of the left hand, and to complete a one-page sun exposure questionnaire from which several indices of ultraviolet radiation exposures were de®ned. Haematoxylin±and-eosin stained sections were assessed for degree of dermal elastosis on a scale from 0 (no evidence of elastotic ®bres) to 6 (severe elastosis). Multiple linear regression models considered the association between ultraviolet radiation exposure indices and dermal elastosis grade. Results: Lifetime sun exposure hours and actinic UVA exposure were both associated with degree of dermal elastosis in a log-linear relationship (p

Regarding the outdoor behavior of the Caucasian population, modern sunscreens should provide high and broad-spectrum ultraviolet protection in the ultraviolet B as well as in the ultraviolet A range and should be photochemically stable for ultraviolet doses, which can be expected in solar radiation. At present an assessment of the photostability of suncare products is not a general requirement before marketing. In order to evaluate the photostability of suncare products we conducted an in vitro test and measured the spectral absorbance of 16 sunscreens before, and after exposure to increasing biologically weighted standard erythema doses (5, 12.5, 25, 50) of solar-simulated radiation. Seven of 16 suncare products showed a significant dose- and wavelength-dependent decrease of the ultraviolet A protective capacity, whereas the ability to absorb ultraviolet B was not affected. In the ultraviolet A range, the decrease of absorbance (photoinactivation), respectively, the increase of transmission was 12-48% for an ultraviolet exposure of 25 standard erythema dose. Photoinactivation started in the wavelength range between 320 and 335 nm with a maximum above 350 nm. Furthermore, our analysis showed that the behavior of suncare products was not predictable from its individual ingredients. Neither complex combinations of organic filters nor addition of inorganic filters could absolutely prevent photoinactivation. The inclusion of a single photounstable filter did not mean photoinstability of the complete suncare product. Photoinactivation of sunscreens appears to be an underestimated hazard to the skin, first, by formation of free radicals, second, by increased ultraviolet A transmission.

Ultraviolet radiation-induced immunosuppression is thought to play a part in skin cancer. Several studies have indicated that sunscreens that are designed to protect against erythema failed to give comparable protection against ultraviolet radiation-induced immunosuppression. One possible reason for this discrepancy is inadequate ultraviolet A protection. This study evaluated the level of immunoprotection in mice afforded by two broad-spectrum sunscreens with the same sun protection factor, but with different ultraviolet A protection factors. Both sunscreens contained the same ultraviolet B and ultraviolet A filters, in the same vehicle, but at different concentrations. Solar simulated radiation dose–response curves for erythema, edema, and systemic suppression of contact hypersensitivity were generated and used to derive protection factors for each end-point. The results of three different techniques for determining immune protection factor were compared. A comparison of the two sunscreens showed that the protection factor for erythema in mice was similar to that determined in humans (sun protection factor) but the protection factor for edema in mice was lower. Both sunscreens protected against suppression of contact hypersensitivity but the product with the higher ultraviolet A-protection factor showed significantly greater protection. The three techniques for determining immunoprotection gave very similar results for a given sunscreen, but immune protection factor was always lower than sun protection factor. These data suggest that sun protection factor may not predict the ability of sunscreens to protect the immune system and that a measure of ultraviolet A protection may also be necessary.

8-Methoxypsoralen plus ultraviolet A light is suggested to shift T lymphocytes from Th2 to Th1 cells. To clarify this issue, we examined the effects of 8-methoxypsoralen/ultraviolet A on the expression/production of cytokines in peripheral blood mononuclear cells from normal subjects and a Sézary syndrome patient. 8-Methoxypsoralen/ultraviolet A augmented the expression of mRNAs for interferon-gamma and interleukin-2 and reduced those for interleukin-4 and interleukin-10. It seems that this enhancement of Th1 cytokines is caused by increment of cytokine production by Th1 cells but not by conversion of Th2 cells to produce Th1 cytokines. The number of interferon-gamma-secreting lymphocytes was markedly increased in 8-methoxypsoralen/ultraviolet A-treated peripheral blood mononuclear cells 20 h after treatment, whereas that of Th2 cytokine-producing cells was decreased. Accordingly, the amount of interferon-gamma was elevated in culture supernatants from 8-methoxypsoralen-phototreated peripheral blood mononuclear cells, whereas interleukin-4 was significantly reduced. This enhanced production of interferon-gamma, however, was found only until 3 d after 8-methoxypsoralen phototreatment and was declined by 5 d after treatment. Finally, 8-methoxypsoralen/ultraviolet A treatment of T cells regulated their ability to induce keratinocyte CD54 expression. Our results show that 8-methoxypsoralen/ultraviolet A has a transient but biologically active Th1-skewing action in human T cells, suggesting that 8-methoxypsoralen/ultraviolet A exerts a beneficial therapeutic effect on Th2-mediated or Th2-malignant diseases.

Ultraviolet light causes both acute and chronic changes in extracellular matrix. We sought to examine the effects of different ultraviolet wavelengths on expression of matrix-related genes in fibroblasts. We previously reported that tropoelastin gene expression in vivo decreases with acute ultraviolet B exposure, and interleukin-1 alpha-mediated upregulation of tropoelastin is blocked in vitro after ultraviolet B radiation. In this study, we found that only ultraviolet B, but not ultraviolet A or ultraviolet A1, blocked the ability of interleukin-1 alpha to stimulate tropoelastin expression in vitro. Ultraviolet B and interleukin-1 alpha synergistically increased tumor necrosis factor-alpha secretion by fibroblasts, a finding not seen with ultraviolet B alone nor with ultraviolet A or ultraviolet A1 combined with interleukin-1 alpha. Keratinocytes showed a similar ultraviolet B-specific induction of tumor necrosis factor-alpha production. Addition of tumor necrosis factor-alpha to cultured fibroblasts blocked interleukin-1 alpha-induced stimulation of tropoelastin message, and addition of anti-tumor necrosis factor-alpha antibodies restored the responsiveness of tropoelastin and collagen messages to exogenous interleukin-1 alpha after ultraviolet B exposure. We conclude that interleukin-1 alpha in combination specifically with ultraviolet B induces fibroblasts to secrete tumor necrosis factor-alpha, and that this ultraviolet B-specific induction of tumor necrosis factor-alpha secretion is responsible for effects of ultraviolet B on the expression of matrix-related genes in the skin.

PUVA (combination of psoralens and ultraviolet A radiation) is a potent inducer of melanogenesis in normal human skin. The molecular mechanisms underlying this effect are poorly characterized. This study was undertaken to investigate the action of PUVA on melanogenesis in S91 murine melanoma cells and in cultured normal human melanocytes. Tyrosinase and DOPAchrome tautomerase (DCT) activities as well as melanin neosynthesis were measured in PUVA-treated pigment cells. To determine whether a correlation exists between PUVA-induced melanogenesis and expression of melanogenic enzymes, we analyzed the levels of tyrosinase, DCT, and tyrosinase-related protein-1 (TRP-1 or gp75) by western blotting in PUVA-treated cells. We demonstrate that UVA upregulates tyrosinase activity and melanin content with 5-methoxypsoralen at 1 microM. This phenomenon depends on the energy delivered during phototreatment. In both human and mouse cells, stimulation of melanogenesis correlated with an increase of the amount of tyrosinase. In PUVA-treated S91 cells, tyrosinase mRNA was increased, but no stimulation of DCT activity occurred in these cells, in agreement with the unchanged amount of DCT protein in cell extracts. On the contrary, in melanocytes treated with PUVA, a decrease in DCT protein was observed. Finally, the amount of TRP-1 protein was not affected by PUVA in either S91 cells or melanocytes. These results show that melanogenesis induced by PUVA is related to an increase in expression of tyrosinase. In melanocytes, melanogenesis and DCT are negatively correlated, which suggests that PUVA favors the metabolic pathway of dark-eumelanins with high UV-protective properties. This study also suggests that PUVA regulates tyrosinase, DCT, and TRP-1 expression in a noncoordinate manner.

Short-term and long-term survival of cultured neonatal foreskin melanocytes from black and white individuals were assessed following a single exposure to simulated sunlight or ultraviolet A (UVA) radiation. Melanocytes from black individuals contained significantly more melanin than melanocytes from white individuals (p less than 0.05). Black and white melanocytes had similar survival profiles following simulated sunlight exposure, whereas black melanocytes were significantly more resistant to UVA cytotoxicity than melanocytes from white subjects (p less than 0.05) at UVA doses above 15 J/cm2. There was no difference in unscheduled DNA synthesis in the black or white melanocytes following simulated sunlight exposure and no unscheduled DNA synthesis was measurable following melanocyte exposure to UVA radiation. Low-dose UVA (1 or 5 J/cm2) was mitogenic to both black and white melanocytes. By analysis of co-variance, the melanin content of melanocytes of black and white subjects was significantly (p less than 0.05) associated with susceptibility to UVA killing; melanocytes with high melanin content had high resistance to UVA cytotoxicity and those with low melanin content had low resistance to UVA cytotoxicity. From these data we suggest that the higher melanin content of melanocytes of black subjects confers increased resistance to UVA damage. This is likely to be of importance in epidermal photodamage.

Calcitriol ointment has been approved for the treatment of psoriasis in many countries around the world. It may be prescribed in conjunction with phototherapy. Our purpose was to evaluate the effect of various therapeutic ultraviolet modalities on the stability of calcitriol and, conversely, to study the effects of calcitriol ointment on transmission of different forms of ultraviolet light. Calcitriol ointment 3 microg per g was irradiated with 10 J per cm2 ultraviolet A, 100 mJ per cm2 broadband ultraviolet B, and 3.0 J per cm2 narrowband ultraviolet B, and its stability was compared with samples exposed to fluorescent light and ambient sunlight. Ultraviolet A and ultraviolet B transmission were measured through thin and thick layers of calcitriol 3 microg per g ointment and vehicle. More than 90% of calcitriol ointment is degraded upon exposure to ultraviolet A, broadband ultraviolet B, and narrowband ultraviolet B. Transmission of ultraviolet A is reduced through calcitriol ointment and its vehicle by 17%-31% and 17%-41%, respectively. Transmission of ultraviolet B is reduced by 67%-87% through vehicle and 50%-83% through calcitriol ointment. When used in conjunction with phototherapy, calcitriol ointment should be applied after ultraviolet exposure, not before. Calcipotriene, the only vitamin D analog already approved for psoriasis in the USA, has been used successfully in combination with ultraviolet B and psoralen plus ultraviolet A.

Previous studies of exposure of normal skin to ultraviolet radiation have demonstrated a cumulative effect lasting greater than 24 h when repeated suberythemal exposures are given. However, the time course of recovery from a single suberythemal dose of ultraviolet A (UVA) or ultraviolet B (UVB) has not been determined. We show here for the first time that the period required for recovery of normal skin (as measured by delayed erythema) following a single suberythemal dose of UVA is between 30 and 48 h, and for UVB is between 24 and 30 h. Photoprotection was noted for both UVA and UVB from the fifth through the ninth day after the single suberythemal exposure, but was only statistically significant on the fourth day after UVB exposure. The curve depicting recovery from a single suberythemal dose of UVA or UVB from the first irradiation time through the fourth day after irradiation may be described as an exponential decay curve. Formulas are given for both UVA and UVB which describe the exponential nature of this curve. These formulas may be used to predict the exact difference in erythema threshold between preirradiated and normal skin. From the fourth day after exposure to the ninth day, the curve is nearly constant. The nature of the recovery curve in the first 4 days after exposure suggests that an exponential decay process occurs in UVA or UVB damage, consistent with unstable photoproduct decay, DNA repair, or constitutive enzymatic processes.

Sunscreens have been advocated as an important means of preventing skin cancer. Ultraviolet radiation induced immunosuppression is recognized as an important event in skin cancer development, yet the effectiveness of sunscreens in protecting the human immune system from ultraviolet radiation (i.e. ultraviolet radiation) is still unclear. The only currently accepted method of sunscreen rating is the sun protection factor system based on the prevention of erythema. We determined immune protection factors for six commercially available sunscreens using a nickel contact hypersensitivity model in humans. Both sun protection factor and immune protection factor testing was performed using the same solar simulated ultraviolet radiation source and dose–responses were used to determine endpoints both with and without sunscreens. We found that the immune protection factor did not correlate with the sun protection factor; however, immune protection factor was significantly correlated to the ultraviolet A protective capability of the sunscreens, indicating that sunscreen protection from ultraviolet A is important for the prevention of ultraviolet immunosuppression, when there is constant ultraviolet B protection. We recommend that sunscreens should be rated against their immune protective capability to provide a better indication of their ability to protect against skin cancer.

It is well established that ultraviolet radiation has immunomodulatory effects that may be involved in skin cancer. Recent studies have shown that ultraviolet A radiation (320-400 nm) as well as ultraviolet B (290-320 nm) is immunosuppressive. This means sunscreens that mainly absorb ultraviolet B (protection against erythema) may be less effective in preventing ultraviolet radiation-induced immunosuppression than broad-spectrum products. We have studied the effects of ultraviolet A exposure on the human delayed-type hypersensitivity response and compared the efficacy of sunscreens having different levels of ultraviolet A protection under both solar-simulated radiation and outdoor real-life solar exposure conditions. Delayed-type hypersensitivity was assessed using recall antigens. In a first study, two groups of volunteers were exposed to ultraviolet A (either full spectrum ultraviolet A or ultraviolet A1) without prior application of sunscreen and they were shown to exhibit significantly reduced delayed-type hypersensitivity responses. In order to compare the efficacy of sunscreens in preventing photoimmunosuppression, three groups of subjects received 10 cumulative exposures to solar-simulated radiation; one group was exposed unprotected and the other two were exposed after being applied either a ultraviolet B or a broad-spectrum sunscreen, each with the same sun protection factor 9, but with different ultraviolet A protection factors 9 and 2. Then, an outdoor study was conducted in which delayed-type hypersensitivity was assessed before and after six daily exposures. Two different groups of subjects were treated with one of two sunscreens having the same sun protection factor 25 but different ultraviolet A-protection factors. In unprotected volunteers, responses to delayed-type hypersensitivity tests were significantly reduced irrespective of ultraviolet exposure conditions (full spectrum ultraviolet A, ultraviolet A1, solar-simulated radiation). The ultraviolet B sunscreen failed to protect from solar- simulated radiation-induced immunosuppression. In contrast, the broad-spectrum sunscreen having the same sun protection factor but providing high protection in the ultraviolet A range significantly reduced local ultraviolet-induced immunosuppression and prevented the distal effects. In the outdoor study, as compared with delayed-type hypersensitivity responses obtained before sun exposure, no alteration of immune response was detected when the skin was protected by broad-spectrum sunscreen sun protection factor 25 and ultraviolet A-protection factor 14. Conversely, a broad-spectrum sunscreen sun protection factor 25 ultraviolet A-protection factor 6 failed to protect against the sun-impaired response. The above studies clearly demonstrate the role of ultraviolet A in the induction of photoimmunosuppression together with the need for sunscreen products providing efficient photoprotection throughout the entire ultraviolet spectrum.

Ultraviolet A radiation from sunlight is a major human health concern, as it is not absorbed by the ozone layer and can deeply penetrate into the skin causing skin damage. To study the molecular mechanism involved in the ultraviolet A effect, human HaCaT keratinocytes were exposed to ultraviolet A at doses of 10 J per cm2 and 30 J per cm2. Ultraviolet A irradiation caused dose- and time-dependent apoptotic cell death, as evidenced by DNA fragmentation, flow cytometry, and the activation of caspase-3. To study the genes altered by ultraviolet A at an apoptosis-inducing dose (30 J per cm2), cells were harvested immediately after ultraviolet A treatment (0 h), and 6 h and 24 h after ultraviolet A exposure. Total RNA was extracted for microarray and real-time RT-PCR analysis, and cellular proteins were extracted for western blot analysis. Of the selected critical genes/proteins, the induction of c-Jun, c-myc, and p33ING1, and the repression of epidermal growth factor receptor, inhibitor of apoptosis protein, and survivin pathways, could be involved in ultraviolet-A-induced apoptosis. On the other hand, the late induction of cyclin D1 and cyclin-dependent kinase 4 was indicative of possible cell cycle recovery in surviving cells. Real-time RT-PCR analysis confirmed these results and a majority of the protein levels paralleled their corresponding RNA levels. In addition, ultraviolet A treatment altered the expression of genes involved in signal transduction, RNA processing, structural proteins, and metabolism in a time-dependent manner. This initial microarray analysis could advance our understanding of cellular responses to ultraviolet A exposure, and provide a platform from which to further study ultraviolet-A-induced apoptosis and carcinogenesis.