Low-dose UV-radiation sensitizes keratinocytes to TRAIL-induced apoptosis

The impact of low-dose ultraviolet light (UV-light) on apoptotic susceptibility of keratinocytes (KCs) induced by TRAIL is unclear. Skin expresses a functional form of TRAIL, and while sun exposure influences TRAIL death receptors, a role for decoy receptors has not been evaluated. Unraveling mechanisms involving apoptotic sensitivity of KCs is important because skin is the first target of UV-light, and a site for commonly occurring cancers. Since apoptosis is a homeostatic process eliminating UV-light induced DNA damaged cells, elucidating molecular events regulating apoptosis enhances understanding of cutaneous photocarcinogenesis. Here we demonstrate low-dose UV-light enhances susceptibility of KCs to TRAIL-induced apoptosis. Low-dose UV-light selectively reduces decoy receptors, without influencing death receptor levels. UV-induced enhanced apoptotic susceptibility was reduced by over-expression of decoy receptor TRAIL-R4, but not TRAIL-R3; or treatment with thiol compound pyrrolidine dithiocarbamate (PDTC), which also enhanced TRAIL-R4 levels. Besides influencing decoy receptors, low-dose UV-light plus TRAIL also synergistically promoted cytochrome c and Smac release from mitochondria. Inhibitors directed against caspases 2, 3, 8, and 9 reduced the synergistic apoptotic response following low-dose UV-light plus TRAIL exposure; as did forced over-expression of Bcl-x and dominant negative (DN) constructs of FADD and caspase 9. Thus, relative levels of decoy receptors significantly influence susceptibility of KCs to TRAIL-induced apoptosis with concomitant low-dose UV-light exposure; in addition to the apoptotic pathway mediated by mitochondrial permeabilization.     

Fragmentation of DNA lymphocytes the human in dynamics of development apoptosis, induced influence of UV-radiation and reactive oxygen species

It is established that UV-light (240-390 nm) in doses of 151, 1510 and 3020 J/m2 and reactive oxygen species and singlet oxygen induce DNA fragmentation lymphocytes cells of the human 20 h after influence. Using a method of DNA-comets it is revealed that DNA damages (single strand breaks) are found out right after UV-irradiations of lymphocytes in doses of 1510 and 3020 J/m2 and additions hydrogen peroxide in concentration of 10-6 mol/l (a comet of type C1) and reach a maximum through 6 h after influence on of cells UV-light and ROS (comets of types C2 and C3). Assumption about the leading part of a p53-dependent way in realization apoptosis human lymphocytes in the conditions of influence of UV-light and reactive oxygen species is put forward.     

Electrical breakdown of erythrocyte membranes during UV-radiation

The electric stability of erythrocyte membrane was shown to reduce under UV-radiation. The disturbing action of UV-radiation depends on the value of membrane potential difference.     

IL-4 suppresses UVB-induced apoptosis in skin

In this study, cutaneous role of IL-4 in UVB-induced apoptosis was investigated using transgenic mice with skin-specific expression of IL-4 (IL-4 Tg mice). The transgenic mice did not show any gross clinical abnormalities. However, epidermis was thickened and increased MHC class II positive cells were detected as well as enhanced expression of inflammatory cytokines such as IL-1 and TNF-alpha in skin. In addition, histological analysis revealed increased infiltration of lymphocytes, acanthosis, hyperkeratosis, and parakeratosis in skin of IL-4 Tg mice. The physiological effect of IL-4 overexpression in skin against environmental stimulus such as UVB was investigated by irradiating wild-type and IL-4 Tg mice with UVB followed by evaluation of apoptosis. The result demonstrated suppressed apoptosis in epidermis of IL-4 Tg mice compared with wild-type mice. To further assess anti-apoptotic function of IL-4 in keratinocytes, stable cell clones were made where IL-4 was constitutively overexpressed and examined for UVB-induced apoptosis. The results showed that apoptosis was remarkably decreased in IL-4 over-expressing cell clones compared with that in mock transfected cells. Collectively, data presented here shows that IL-4 has an inhibitory effect against UVB-induced apoptosis in keratinocytes, suggesting that IL-4 may be an important regulator in cutaneous immunity against UVB.     

umuC-dependent and umuC-independent gamma- and UV-radiation mutagenesis in Escherichia coli

The effects of the umuC36 and umuC122::Tn 5 mutations on gamma- and UV radiation mutagenesis (nonsense, missense, and frameshift mutation assays) in Escherichia coli K12 were studied. Although both mutations reduced radiation mutagenesis, the umuC36 mutation appeared to be leaky since considerably more UV radiation mutagenesis could be detected in the umuC36 strain than in the umuC122::Tn 5 strain. In general, the umuC strain showed much larger deficiencies in UV radiation mutagenesis than they did for gamma-radiation mutagenesis. The mutability of the umuC122:: Tn 5 strain varied depending on the radiation dose, and the mutation assay used. For gamma-radiation mutagenesis, the deficiency varied from no deficiency to a 50-fold deficiency; for UV radiation mutagenesis, the deficiency varied from 100-fold to at least 5000-fold. We concluded that both umuC-dependent and umuC-independent modes function for gamma-radiation mutagenesis, while UV radiation mutagenesis seems to depend almost exclusively on the umuC-dependent mode.     

Mad dogs,Englishmen and apoptosis: the role of cell death in UV-induced skin cancer

Apoptosis plays a critical role in the development and progression of ultraviolet-induced skin cancers. In particular, Fas and Fas ligand (FasL) interactions are known to control the development of sunburn cells or apoptotic keratinocytes in the UV-exposed epidermis. In the absence of functional Fas/FasL signaling, UV-induced apoptosis is diminished and mutations rapidly accumulate. UV-induced suppression of host immunity, a process regulating skin cancer outgrowth, is also controlled through Fas/FasL interactions. Other death receptors, such as the receptor for tumor necrosis factor, may also contribute to UV-induced carcinogenesis and progression. Understanding the involvement of cell death in cancers caused by exposure to sunlight may provide novel approaches for prevention and therapy of these ever-increasing malignancies.     

Seasonality of UV-radiation and vitamin D status at 69 degrees north.

The main purpose with this study was to assess the seasonal variation in measured UV-radiation and its impact on vitamin D status throughout one year in subjects living at high latitude. Blood samples drawn from 60 volunteers (44 women, 16 men) living at Andenes (69 degrees N), Norway, were collected throughout one year, at two-month intervals. The blood samples were analysed for 25-hydroxy vitamin D [25(OH)D]. Data on dietary intakes of vitamin D, time spent in daylight, use of sun beds and sun seeking holidays were collected by using questionnaires. The ambient vitamin D effective UV-radiation was measured at a site near by Andenes, and the number of hours spent outdoors with sufficient radiation for cutaneous vitamin D production (UV-hours) was estimated for each day. The mean 25(OH)D values were significantly higher at the end of the summer and in December, 2004 and varied from 42.0 nmol L(-1) in October, 2004 and April, 2005 to around 47 nmol L(-1) in December, 2004 and September, 2005. For the whole group, a positive relationship between UV-hours and 25(OH)D was found at UV-hours>or=3.5. However, for subjects with lower 25(OH)D levels i.e. at least one blood measurement with 25(OH)D<37.5 nmol L(-1), the positive relationship were found at around 1.5 UV-hours and more, whereas for the group of subjects that had all their vitamin D values above 37.5 nmol L(-1), positive relationship was found at UV-hours>or=4.0, when adjusting for vitamin D intake, sun bed use and sun seeking holidays. The generally high dietary intakes of vitamin D, especially in winter, mask largely the effect of seasonal variation in UV-exposure, causing an atypical seasonal variation in vitamin D status. The UV-hour variable significantly predicted 25(OH)D levels in blood when adjusted for intakes and artificial UV-radiation exposure and sun holidays abroad.     

Ha-Ras sensitizes transformed mouse skin cells to Anisomycin-induced apoptosis

Efforts have been made to develop a chemoprevention that selectively triggers apoptosis in malignant cancer cells. Here, we demonstrated that a mutated Ha-Ras activity is required in Anisomycin-induced apoptosis in transformed keratinocytes. Anisomycin stimulates JNK activity and apoptosis in oncogenic Ha-Ras positive cells, but not in normal keratinocytes. This effect was demonstrated in stably transfected cells with dominant negative Ha-Ras, that protected transformed cells, and oncogenic Ha-Ras that sensitized non-transformed cells to Anisomycin-induced apoptosis. Lastly, the treatment of cells with inhibitors of the JNK displayed resistance to Anisomycin induced apoptosis. These data suggests that the oncogenic Ha-Ras is important for Anisomycin-induced JNK activation and apoptosis in transformed keratinocytes.     

ESR spectra of paramagnetic centers of ferricytochrome c, induced by UV-radiation at 77 K

Paramagnetic properties of ferricytochrome c aqueous glyceric solutions exposed to UV-light of various wave lengths at 77K have been investigated. Absorption of longwave UV- and visible light by hemin (5-240 min) does not induce the formation of paramagnetic centres in hemoprotein. However absorption of UV-light by chromophorms of a protein component of ferricytochrome c induces the formation of free radicals with a quantum yield of 0.07. During long-term (40 to 200 min) UV-irradiation (260-380 nm) of protein solutions several types of paramagnetic centres including HCO radicals are formed.     

Potentiation of UVB-induced apoptosis by novel phytosphingosine derivative, tetraacetyl phytosphingosine in HaCaT cell and mouse skin

Inappropriate apoptosis results in the epidermal hyperplasia as in psoriasis and UVB irradiation has been successfully used to treat this kind of skin disorders. Previously, we reported that the novel phytosphingosine derivative, tetraacetyl phytosphingosine (TAPS) induced apoptosis in HaCaT cells. This study examined the effect of UVB irradiation and/or TAPS on the induction of apoptosis in HaCaT. 10 mJ/cm2 of UVB irradiation or 10 microM of TAPS alone exhibited weak cytotoxicity but co-treatment of UVB and TAPS synergistically enhanced the cytotoxicity and apoptosis in HaCaT. The cells treated with UVB and TAPS showed much higher levels of cleaved caspase-3, -8, -9 and Bax than with UVB or TAPS alone, whereas Bcl-2 level was decreased by co-administration of UVB and TAPS. In hairless mice, co-treatment of UVB and TAPS synergistically increased apoptosis, as shown in the HaCaT co-treated with UVB and TAPS. Furthermore, UVB irradiation caused an increase of apoptotic cells in the epidermis and the TAPS-treated mice showed an increase of apoptotic cells in the dermis as well as in the epidermis. These results suggest that the TAPS co-treatment synergistically increases the level of UVB-induced apoptosis via caspase activation by regulating the level of pro-apoptotic Bax and anti-apoptotic Bcl-2.     

Prognosis of yeast cells recovery after simultaneous exposure to UV-radiation and hyperthermia

The results of experimental investigations of survival of diploid yeast cells Saccharomyces cerevisiae (strain XS800) after simultaneous exposure to UV-radiation (254 nm) and hyperthermia (53-57 degrees C) have been described. It was shown that the portion of cells capable of recovery in innutrient medium after the action of these agents decreased with the increasing of temperature under which the irradiation was occurred. Mathematical model taking into account the synergistic interaction was suggested for quantitative prediction of irreversible component after combined actions of these agents. A good correspondence between experimental data and model predictions has been demonstrated. The importance of the results obtained for the interpretation of the synergistic interaction mechanisms are discussed.     

Wavelength,dose, skin type and skin model related radical formation in skin

The exposure to sun radiation is indispensable to our health; however, a long-term and high exposure could lead to cell damage, erythema, premature skin aging, and promotion of skin tumors. An underlying pathomechanism is the formation of free radicals which may induce oxidative stress at elevated concentrations. Different skin models, such as porcine-, murine-, human- ex vivo skin, reconstructed human skin (RHS) and human skin in vivo, were investigated during and after irradiation using X- and L-band EPR spectroscopy within different spectral regions (UVC to NIR). The amount of radical formation was quantified with the spin probe PCA and the radical types were measured ex vivo with the spin trap DMPO. The radiation dose influences the types of radicals formed in the skin. While reactive oxygen species (ROS) are always pronounced at low doses, there is an increase in lipid oxygen species (LOS) at high doses. Furthermore, the radical types arise independent from the irradiation wavelength, whereas the general amount of radical formation differs with the irradiation wavelength. Heat pre-stressed porcine skin already starts with higher LOS values. Thus, the radical type ratio might be an indicator of stress and the reversal of ROS/LOS constitutes the point where positive stress turns into negative stress.Compared to light skin types, darker types produce less radicals in the ultraviolet, similar amounts in the visible and higher ones in the infrared spectral region, rendering skin type-specific sun protection a necessity.     

Culture age impacts Plectosporium alismatis propagule yields and subsequent desiccation and UV-radiation tolerance

The effect of culture age on yields, desiccation tolerance and resistance to ultraviolet radiation of Plectosporium alismatis, a potential mycoherbistat of aquatic weeds in Australian rice fields, was studied. P. alismatis was grown in a liquid basal medium supplemented with malt extract and sodium nitrate and harvested after 7, 14 or 21 days incubation. Although chlamydospore yields harvested from 14-day-old liquid cultures were significantly higher (29.2×10⁵ chlamydospores mL^?1) than chlamydospore yields harvested from 7-day-old liquid cultures (1.07×10⁵ chlamydospores mL^?1) or from 21-day-old liquid cultures, the germination of freshly-harvested chlamydospores from 7-day-old cultures (72.7%) was significantly reduced when propagules were grown for 14 days (55.3%). When exposed to UV-radiation, conidia and chlamydospores harvested from 14-day-old cultures germinated at a lower rate (<20%) than conidia and chlamydospores harvested from 7-day-old cultures (>40%). When conidia and chlamydospores were dried and subsequently exposed to UV, less than 30% of propagules harvested from 7-day-old cultures germinated, whereas less than 10% of propagules harvested from 14-day-old cultures germinated. A three-way analysis of variance including culture age, UV exposure and type of propagules confirmed that the culture age had more impact on the germination of fresh or dry propagules (P=0.00001 and P=0.0004, respectively) than the type of propagules considered (P=0.5). These results demonstrate that the culture age impacts significantly propagule yields and germination of P. alismatis conidia and chlamydospores, particularly after stress caused by dehydration and/or exposure to UV-B radiation.     

TIG3 tumor suppressor-dependent organelle redistribution and apoptosis in skin cancer cells

TIG3 is a tumor suppressor protein that limits keratinocyte survival during normal differentiation. It is also important in cancer, as TIG3 level is reduced in tumors and in skin cancer cell lines, suggesting that loss of expression may be required for cancer cell survival. An important goal is identifying how TIG3 limits cell survival. In the present study we show that TIG3 expression in epidermal squamous cell carcinoma SCC-13 cells reduces cell proliferation and promotes morphological and biochemical apoptosis. To identify the mechanism that drives these changes, we demonstrate that TIG3 localizes near the centrosome and that pericentrosomal accumulation of TIG3 alters microtubule and microfilament organization and organelle distribution. Organelle accumulation at the centrosome is a hallmark of apoptosis and we demonstrate that TIG3 promotes pericentrosomal organelle accumulation. These changes are associated with reduced cyclin D1, cyclin E and cyclin A, and increased p21 level. In addition, Bax level is increased and Bcl-XL level is reduced, and cleavage of procaspase 3, procaspase 9 and PARP is enhanced. We propose that pericentrosomal localization of TIG3 is a key event that results in microtubule and microfilament redistribution and pericentrosomal organelle clustering and that leads to cancer cell apoptosis.     

Resveratrol induces apoptosis involving mitochondrial pathways in mouse skin tumorigenesis

Resveratrol, a plant constituent enriched in the skin of grapes, is one of the most promising agents for chemoprevention. In the present study, resveratrol-induced apoptosis in 7, 12-dimethylbenz[a]anthracene (DMBA)-initiated and 12-O-tetradecanoylphorbol-13-acetate (TPA) promoted, mouse skin tumors. The chemopreventive effects of resveratrol in terms of delayed onset of tumorigenesis, cumulative number of tumors and average number of tumors/mouse were recorded. Resveratrol treatment resulted in regression of tumors (28%) after withdrawal of the TPA treatment. Induction of apoptosis by resveratrol in DMBA-TPA induced skin tumors was recorded by the appearance of a sub-G1 fraction (30%) using flow cytometry and an increase in the number of apoptotic cells by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay. Western blot analysis combined with multivariable flow cytometry, showed that resveratrol application induces the expression of the p53 and pro-apoptotic Bax, with concomitant decrease in anti-apoptotic protein Bcl-2. Alteration in Bax/Bcl2 ratio by resveratrol treatment resulted in apoptosis, which was associated with the release of cytochrome c and induction of apoptotic protease-activating factor-1(APAF-1). Further, this effect was found to result in cleaved fragments of caspase-9,-3, and poly (ADP-ribose) polymerase (PARP). These findings demonstrate for the first time that resveratrol induces apoptosis through activation of p53 activity in mouse skin tumors, thereupon suggesting its chemopreventive activity, through the modulation of proteins involved in mitochondrial pathway of apoptosis.     

Characterization of primary culture of rainbow trout (Oncorhynchus mykiss) skin explants: Growth, cell composition, proliferation, and apoptosis

A trout (Oncorhynchus mykiss) epidermal skin primary explant system was evaluated over 8 d by light and electron microscopy. Three distinct regions of the explant outgrowth were identified on the basis of cell composition. The area immediately adjacent to the founder tissue contained mainly small migrating cells and mucous cells. Of the former. about 20% were mitotic and 6% apoptotic. The middle area was characterized by differentiated pavement cells and mucous cells, with fewer small migrating cells. Proliferation was approximately 30% and apoptosis 5%. Over time, total cell numbers halved as more pavement cells differentiated. The growing front contained many mucous and small migrating cells initially, with few pavement cells. About 50% of the cells were in the proliferative phase, and 5% were apoptotic. Later, there were fewer migrating and mucous cells, with a higher number of pavement cells. About 9% of the cells were apoptotic, and 70% of the cells were proliferating. As in vivo, pavement cells had apical microridges, although they were vacuolated and contained phagocytosed apoptotic bodies. The data and observations are based on the numbers of cell cultures prepared from separate trout giving the sample size n = 7. As this culture system is reproducible and closely approximates the epidermis of trout, it is a powerful tool to study the effects of pollutants, parasites, and endocrine factors on fish skin, eliminating whole-animal factors and reducing the number of experimental animals required.