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Biomedical subjects

V DeLeo

Publications and source records attributed to V DeLeo.

7 recordsLinked to original sources

Long-wave ultraviolet light induces phospholipase activation in cultured human epidermal keratinocytes.

Long wave ultraviolet radiation (UVA) has been shown to play an important role in the overall response of skin to solar radiation, including sunburn, tanning, premature aging, and non-melanoma skin cancer. UVA induction of inflammation in human skin is thought to be mediated by membrane lipid derived products. In order to investigate the mechanism of this response we examined the effect of UVA on phospholipid metabolism of human epidermal keratinocytes in culture. Keratinocytes were grown in serum free low calcium medium. The cells were prelabeled with [3H] arachidonic acid or [3H] choline and irradiated with UVA (Honle 2002-Hg vapor lamp). Identification and quantitation of specific membrane phospholipid-derived components was achieved using high-performance liquid chromatography, paper chromatography, and radioimmunoassay. UVA resulted in a linear dose dependent release of [3H] arachidonic acid into medium between 1 and 20 joule/cm2. This response was inhibited in an oxygen-reduced environment. The radiolabel released was predominantly free arachidonate and cyclooxygenase metabolites. Cyclooxygenase metabolites prostaglandin E2 and prostacyclin derivative, 6-keto-prostaglandin F1a, were stimulated following UVA irradiation, but the lipoxygenase metabolite, leukotriene B was not detected. Maximal release was measured immediately after irradiation and changed little over 24 h post-irradiation. UVA stimulated an increase of [3H] choline metabolites glycerophosphorylcholine and phosphorylcholine in media extracts suggesting UVA activation of phospholipase C and phospholipase A2 or diacylglyceride lipase.

Arachidonic Acid

Ultraviolet-B (290-320 nm)-irradiation inhibits epidermal growth-factor binding to mammalian cells.

Mitogens, such as polypeptide growth factors and phorbol ester tumor promoters, act by binding to specific receptors and inducing a pleiotropic response in cultured mammalian cells, which results in the induction of cellular proliferation. An early effect of such agents is the inhibition of binding of epidermal growth factor (EGF) to its receptor. Ultraviolet radiation has also been shown to induce a proliferative response in vivo and in vitro and to act as a tumor promoter in animal skin. We, therefore, examined the effect of ultraviolet radiation (UVB - 290-320 nm) on EGF binding to cells in culture. We found that UVB (100-300 J/m2) induced a rapid, dose-dependent inhibition of EGF binding in a mouse fibroblast cell line, which resulted from a decrease in both number and affinity of binding sites. Phosphorylation of the EGF receptor by protein kinase C (PKC) is not likely to be the mechanism for inhibition, since UVB treatment did not result in PKC activation or modulation of phorbol diester binding.

Animals

Photoallergy to benzophenone.

Incorrect diagnosis of photoallergy to sunscreen products represents a unique clinical dilemma. Increasing sunscreen usage for suspected idiopathic photosensitivity or a change to a sunscreen containing the same photoallergen only worsens the problem. While photoallergy to p-aminobenzoic acid and its esters is well known by dermatologists and the lay public, benzophenone photoallergy is not well appreciated. We report herein the cases of four individuals with photoallergy to oxybenzone in sunscreens. It is likely that such reactions will become more commonplace since oxybenzone is by far the most frequently used agent in modern, high sun protection factor sunscreens (greater than 8 sun protection factor) being marketed today.

4-Aminobenzoic Acid

Ultraviolet radiation alters choline phospholipid metabolism in human keratinocytes.

Ultraviolet radiation B (UVB-290-320 nm) induces inflammation and hyperproliferation in human epidermis. This response is associated with the recovery from irradiated skin of inflammatory mediators derived from membrane phospholipids. We have previously reported that UVB stimulates the production of such mediators by human keratinocytes (HK) in culture. In these studies we examined the effect of UVB on the metabolism of choline containing phospholipids in HK prelabeled with [3H] choline. UVB (400-1600J/m2) stimulated a dose dependent release of [3H] choline from HK within minutes of irradiation. Examination of media extracts by paper chromatography revealed that the released [3H] choline was predominately in the form of glycerophosphorylcholine. Examination of label remaining in membranes of cells after irradiation by acid precipitation and HPLC revealed that the origin of the released [3H] choline was the membrane phosphatidylcholine/lysophosphatidylcholine. These data support a concept of UVB stimulation of both a phospholipase A (1 or 2) and a lysophospholipase. These UVB induced alterations of HK membrane phospholipid metabolism likely have profound effects on UVB-induced inflammation and control of cell growth in human skin.

Cells, Cultured

Immunological detection and visualization of 8-methoxypsoralen-DNA photoadducts.

Monoclonal antibodies specific for DNA damaged by 8-methoxypsoralen (8-MOP) plus ultraviolet A (UVA) light were used to study adduct formation in human keratinocytes and mouse and rat skin in vivo. This antibody does not cross-react with nonmodified DNA or free 8-MOP. Sensitive competitive enzyme-linked immunosorbent assays with color or fluorescence endpoints were used to quantitate adducts on DNA isolated from treated keratinocytes or skin samples. Localization of 8-MOP-DNA adducts was studied by indirect immunofluorescence with fluorescein-conjugated anti-mouse-IgG antibodies. When cultured keratinocytes were treated with 8-MOP and UVA, immunofluorescence was localized in the nucleus. There was no fluorescence in untreated control cells or treated cells incubated with nonspecific serum. Comparison of intensity of immunofluorescence staining with quantitation of adduct levels by enzyme-linked immunosorbent assay indicated that the limit of sensitivity of the immunofluorescence technique is 9.0 fmol adduct/micrograms DNA or 2.9 adducts/10(6) nucleotides.

Animals

Allergic contact dermatitis from bacitracin.

Bacitracin is a commonly used topical antibiotic. Although occasional reports of adverse reactions to bacitracin have appeared in the medical literature, it is considered to be a nonsensitizing agent by most dermatologists. A description of two patients who demonstrated allergic contact dermatitis from bacitracin is presented, with a review of the pertinent literature.

Adult

Hydroa vacciniforme.

Two patients with hydroa vacciniforme, a rare photodermatosis of unknown etiology, demonstrated distinctive scarring and vesiculobullous skin lesions on light-exposed body areas. Results of blood and urine porphyrin studies were normal, and no systemic abnormalities were noted. A small bullous lesion was produced in normal skin in case 1 with 15 times the minimal erythema dose of ultraviolet energy. The conditions of both patients improved while they were taking beta carotene orally.

Carotenoids