Temperature dependence of photochemical fluorescence fading in Skh-1 hairless mouse collagen.

Background: Type I mammalian collagens have several photolabile fluorescent moieties that absorb UV rays capable of reaching the dermis. We studied the temperature dependence of fluorescence fading as a marker of photochemical damage.

Methods: Collagen solutions were exposed to radiation from 0 to 240 min from either a UVG-11 hand lamp, total dose=1.

Fluorescence of putative chromophores in Skh-1 and citrate-soluble calf skin collagens.

Background/purpose: Fluorescence of Skh-1 hairless mouse and calf skin acid-soluble type I collagens are envelopes of several bands putatively due to tyrosine (excitation/emission peak at 275/300 nm), dihydroxyphenylalanine (dopa; 280/325 nm), tyrosine aggregate (285/360 nm), dityrosine 325/400 nm), and advanced glycation end (AGE) product (370/450 nm), respectively. As these fluorophores can be markers of pathological conditions, we wish to present further evidence for or against these assignments.

Methods: We analysed intact type I mouse collagens and AGE by conventional fluorescence spectroscopy, synchronous spectroscopy, high-performance liquid chromatography (HPLC), and borate quenching.

Temperature dependence of collagen fluorescence.

Dermal collagens have several fluorescent moieties in the UV and visible spectral regions that may serve as molecular probes of collagen. We studied the temperature-dependence of a commercial calf skin collagen and acid-extracted Skh-1 hairless mouse collagen at temperatures from 9 degrees C to 60 degrees C for excitation/emission wavelengths 270/305 nm (tyrosine), 270/360 nm (excimer-like aggregated species), 325/400 nm (dityrosine) and 370/450 nm (glycation adduct). L-tyrosine (1 x 10(-5) M in 0.

Modest in vivo exposure to solar wavelengths induces a visible absorbing chromophore in Skh-1 mouse epidermis.

Background/purpose: In the previous work, we correlated epidermal hyperplasia with increased epidermal absorption in the 250-400 nm region. During a recent review of that work, the apparent formation of a chromophore, with absorption slightly longer than 400 nm, in the epidermis of irradiated animals was noted. In this study, we have extended the transmission measurement to include the 250-800 nm region.

Clothing as solar radiation protection.

The sun is essential for life. Yet, sunlight can also be a source of such deleterious effects as sunburn, and suntanning, as well as premalignant and malignant lesions. These may all occur in individuals with normal responses to sunlight.

Effect of UV on the susceptibility of acid-soluble Skh-1 hairless mouse collagen to collagenase.

Background/purpose: Photoaging of the skin is a result of chronic exposure to environmental ultraviolet radiation (UV). The milieu provided by the extracellular matrix, which significantly influences the behaviour of resident fibroblasts, depends critically on the supermolecular collagen structure. We ask whether direct photochemical treatment of type I collagen with solar wavelengths capable of reaching the dermis can modify the substrate's susceptibility to collagenase in a model in vitro system.

High performance liquid chromatographic assay for the quantitation of total glutathione in plasma.

A simple and widely used homocysteine HPLC procedure was applied for the HPLC identification and quantitation of glutathione in plasma. The method, which utilizes SBDF as a derivatizing agent utilizes only 50 microl of sample volume. Linear quantitative response curve was generated for glutathione over a concentration range of 0.

Effect of UV irradiation on type I collagen fibril formation in neutral collagen solutions.

Background: Collagens have the well-known ability to spontaneously self-associate to form fibrils at physiological temperature and neutral pH in vitro and in vivo. Because solar UV may photochemically alter collagen, the kinetics of fibril formation may be modified. Thus, we have begun a systematic study of the effect of various UV wavebands on fibril formation.

Protection against photodynamic therapy (PDT)-induced photosensitivity by fabric materials.

"Special" highly protective fabrics are now available that offer broad-spectrum protection in preventing sunburn, and possibly other types of photodamage. It is important to know to what extent these fabrics are capable of protecting the wearer against skin cancer, photosensitivity disorders, and inadvertent phototoxic reactions from photodynamic therapy (PDT). We assess the ability of one such special (Solumbra) fabric and one "typical" summer fabric to provide protection against PDT phototoxicity produced in tape-stripped Sk-1 hairless mice by topical 5-aminolevulinic acid (ALA) and (primarily) visible light (360-800 nm).

Photoprotection of mammalian acid-soluble collagen by cuttlefish sepia melanin in vitro.

Several important clinical conditions can result in close association between the pigment melanin and dermal collagen. Because melanin and its precursors can be chemically reactive in ground and excited states, it is important to know whether the resulting melanin-collagen interaction results in photoprotection or photoaggression. Acidic and neutral air-saturated collagen suspensions (0.

Electron transfer and photoprotective properties of melanins in solution.

The polyquinoid nature of eumelanin(s) enables them to couple oxidation of electron donors with the reduction of electron acceptors. We have studied the ability of synthetic (Sigma) and "biological" (cuttlefish sepia) melanins to mediate electron transfer between hydroxybenzene donors (tyrosine, dopa, chemical depigmenters) and model acceptors (ferricyanide, tyrosinase). 1) Depending on the reductant, melanin either retards or accelerates ferricyanide reduction.

Chronic exposure of Sk-1 hairless mice to narrow-band ultraviolet A (320-355 nm).

Several recent investigations collectively suggest that the role of ultraviolet A (UVA) in chronic actinic skin damage may be greater than originally thought. In the present work, the output of a xenon-arc solar-simulator passed through a Bausch & Lomb monochromator in conjunction with a 2-mm Schott WG-320 filter produced narrow-band UVA centered at 338 nm, half-band width 24 nm, I0 = 3.4 +/- 0.

Photochemistry of type I acid-soluble calf skin collagen: dependence on excitation wavelength.

Although previous studies have demonstrated that the predominant photochemistry of type I collagen under 254 nm irradiation may be attributed either to direct absorption by tyrosine/phenylalanine or to peptide bonds, direct collagen photochemistry via solar UV wavelengths is much more likely to involve several age- and tissue-related photolabile collagen fluorophores that absorb in the latter region. In this study, we compare and contrast results obtained from irradiation of a commercial preparation of acid-soluble calf skin type I collagen in solution with UVC (primarily 254 nm), UVA (335-400 nm) and broad-band solar-simulating radiation (SSR; 290-400 nm). Excitation spectroscopy and analysis of photochemically induced disappearance of fluorescence (fluorescence fading) indicates that this preparation has at least four photolabile fluorescent chromophores.

Protection against UV photocarcinogenesis by fabric materials.

Background: Clothing fabrics have long been considered effective protection against short-term and long-term sun damage. Recently, special "highly UV-protective" fabrics have been developed specifically for photosensitive patients.

Objective: To determine if one such fabric will protect hairless mice against (pre)malignant lesions under conditions that will produce skin cancers through a typical summer fabric of moderate sun protection factor (SPF).

In vivo depigmentation by hydroxybenzene derivatives.

Certain mono- and dihydroxybenzene derivatives are selectively cytotoxic for melanocytes in vivo, and can cause depigmentation of skin and hair. We produced selective melanocytotoxicity/hair depigmentation in C57Bl mice by injection of 0.032-1.

Dose-response of chronic ultraviolet exposure on epidermal forward scattering-absorption in SK-1 hairless mouse skin.

This work provides a dose-response model of UV-induced epidermal-stratum corneum thickening induced by irradiation at wavelength lambda. This model assumes that photobiochemical reaction(s) can give rise to hyperplasia in a manner which is predictable from a simple photochemical kinetic scheme. In this work, we derive an equation which predicts an approximately linear relationship between the logarithm of the increase in optical skin thickening measured at 320 nm (delta OD320) and total cumulative dose (DT) seen by the target cells in or near the basal layer.

Melanin accelerates the tyrosinase-catalyzed oxygenation of p-hydroxyanisole (MMEH).

Although pigment melanin has long been though of as "inert," recent work has attested to its chemical reactivity. In this communication, we report that either commercial synthetic melanin prepared by persulfate oxidation of tyrosine ("Sigma melanin") or sepia melanin extracted from cuttlefish markedly accelerates the in vitro oxygenation of p-hydroxyanisole (MMEH), catalyzed by mushroom or B-16 melanoma tyrosinase. Kinetics of 4-methoxy-1,2-benzoquinone formation (lambda max = 413 nm) or of molecular O2 uptake were biphasic, with an initial slow rate ("lag time") followed by a fast linear increase.

Ultraviolet radiation-induced suppression of contact hypersensitivity in relation to padimate O and oxybenzone.

Contact hypersensitivity (CHS) in mice can be induced by cutaneous sensitization followed by elicitation via ear-painting with trinitrochlorobenzene (TNCB). This CHS reaction is systemic and can be suppressed by exposure of mice to suberythemogenic doses of 280-315 nm radiation. In this study, we investigated whether a commercially available water-resistant sunscreen, either SPF-6 or SPF-15, containing Padimate O (UVB absorber) and oxybenzone (UVA absorber), was effective in preventing systemic suppression of CHS induced by either FS36 sunlamp exposure or solar simulating radiation.

Interaction of several mono- and dihydroxybenzene derivatives of various depigmenting potencies with L-3,4-dihydroxyphenylalanine-melanin.

Certain mono- and dihydroxybenzene derivatives cause depigmentation of skin and hair, and appear to be selectively cytotoxic for melanized pigment cells. As direct physical and/or chemical interaction between depigmenter (DP) and pigment melanin may play a role in depigmentation, we have carried out preliminary studies in model systems where such interactions may easily be separated from effects due to tyrosinase, melanosomal proteins, and other components. We have used synthetic L-3,4-hydroxyphenylalanine (L-DOPA)-melanin as a protein-free model pigment and potassium ferricyanide as a model electron acceptor.

Psoralens: a search for more effective derivatives for photochemotherapeutic regimens.

The objective of our studies under the National Toxicology Program on psoralens was to evaluate a new furocoumarin derivative that would be highly efficacious and yet possess little or no systemic toxicity while also having a short effective biologic half-life. In addition, this work allowed for the development of a test system for compound evaluation of various psoralen derivatives. A guinea pig model was used first, followed by definitive studies in hairless mice for evaluation of the phototoxic potentialities of 32 furocoumarins and 4 benzofuran derivatives.

Effect of varying dose of UV radiation on mammalian skin: simulation of decreasing stratospheric ozone.

To better understand the dependence of the incidence of squamous cell carcinoma on changes in solar spectral distribution and dose regimen, we exposed SK-1 hairless mice to solar-simulating radiation (290-400 nm). Selective UV filtration was accomplished by passing this radiation through Schott WG-320 cutoff filters of 0, 0.5, 1.

The rapid induction of cancers in the hairless mouse utilizing the principle of photoaugmentation.

We report a method for rapidly inducing cancer in the hairless mouse utilizing regimen in which an exposure to highly erythemogenic, but otherwise clinically noninjurious, dose of broad spectrum (290-400 nm) ultraviolet light is increased by 20% every 6th day. Clinical and histological observations reveal the presence of squamous cell cancer after as little as 18 days of irradiation. The rate of cancer induction is enhanced by the 320-400 nm component and this enhancement is shown to be a photoaugmentative effect.