Evidence for the existence of a self-regulated enzymatic process within the human stratum corneum -an unexpected role for urocanic acid.

The existence of a flux of proton donors from skin (inner part of the forearm) to the electrode was observed in 12 male and female volunteers. This flux was used to collect and identify the ionic species responsible for skin acidity. It was then found that: (i) pK of these proton donors (pK = 6.13 +/- 0.07) was quasi-identical to that of trans-urocanic acid (6.10), and (ii) the amount of urocanic acid present in stratum corneum was sufficient in itself to explain the acidic level as measured with pH meter (R = 0.8484, n = 10, p = 0.00136). As a result, the contribution of other ionic species can be considered as negligible in normal human skin. The data recorded led us to identify three groups (Fast, Medium, and Slow) characterized by different skin surface pH values (low, medium, and close to neutral) and showing a pH gradient in the outer layers of the stratum corneum, or not. Data analysis suggests that these characteristics depend on urocanic acid production rate within the stratum corneum and that this production rate is self-regulated by its urocanic acid content.

Direct comparison of DNA damage, isomerization of urocanic acid and edema in the mouse produced by three commonly used artificial UV light sources.

Exposure to sunlight can result in a number of harmful effects, including sunburn, erythema, premature aging of the skin, immune suppression and skin cancer. Studies designed to understand the underlying mechanisms often depend upon the use of artificial sources of UV radiation. Unfortunately, conclusions from different laboratories using different lamps often conflict, and it is entirely possible that the different spectra of sunlights used in each may be a source of conflict. To minimize confounding variables, we employed two of the more commonly used UV light sources, fluorescent sunlamps, such as the FS-40 and Kodacel-filtered FS-40 sunlamps, and a xenon arc solar simulator and compared, in one series of standardized experiments, the effects of each light source on DNA damage, urocanic acid isomerization and edema formation. The dose-response curves, calculated by linear regression or curve fitting were compared. The data indicate that DNA damage and urocanic acid isomerization were more sensitive to shorter wavelengths of UV than longer wavelengths, and the biological endpoint of edema most closely correlated with the induction of DNA damage. The results emphasize the dominance of shorter wavelengths within the UV spectrum in damaging biological tissues, even when the solar simulator, which contains significant amounts of UVA, was used and demonstrate that each light source has a characteristic pattern of induction of biochemical and biological endpoints.

Personality correlates of +/- pindolol induced decreases in prolactin.

The present study was conducted to investigate if personality traits can be usefully related to serotonergic vs. dopaminergic action of the 5-HT1a-antagonist +/- pindolol. Forty healthy male volunteers (aged between 20 and 30 years) were randomly assigned to a placebo or a +/- pindolol (30 mg, oral dose) group in a double blind trial. Blood samples were drawn and analyzed for PRL concentrations. In addition, the subjects completed questionnaires on personality. The results indicated that +/- pindolol decreases PRL concentrations depending on personality. While subjects high on impulsivity and related traits (aggression and disinhibition) show lower PRL decreases, well-being and personality traits frequently related to dopaminergic activity were not correlated with changes in PRL. Since reduced (blunted) PRL-responses after 5-HT challenge tests have been reported for impulsives, the present results favor the involvement of primarily serotonergic and probably only secondarily dopaminergic control of +/- pindolol induced PRL decreases.

The use of diethylaminoethyl-cellulose-membrane filters in a bioassay to quantify melanin synthesis.

Melanins are a class of extremely insoluble granular biopolymers with a yet ill-defined chemical structure, properties which render the isolation and quantification of these molecules very difficult. Based on their strong anionic character, however, a biophysical property shared by all melanins, we have developed a method allowing total recovery of the pigment. Soluble melanin is bound and granular melanin retained by DEAE-cellulose membrane filters. This approach provides an excellent means to quantify melanin synthesis in normal human melanocytes alone and in coculture with normal human keratinocytes.

Sunscreens with broad-spectrum absorption decrease the trans to cis photoisomerization of urocanic acid in the human stratum corneum after multiple UV light exposures.

The trans to cis photoisomerization of urocanic acid (UCA) in skin is considered to play an important role in the mechanism of immunosuppression. We have investigated the effects of skin type and various sunscreens with low sun protection factor (SPF) on the UV-induced cis-UCA formation in human skin after exposure to artificial UV light. The rate of cis-UCA formation depends little on the skin type and is reduced by topical application of sunscreens. The rate of cis-UCA formation decreases with increasing SPF and only broad-spectrum, highly protective sunscreens offer protection against the UV-induced formation of cis-UCA, which accumulates in the stratum corneum after multiple UV exposures. A theoretical approach to estimate the distribution of cis-UCA after irradiation indicates that this compound may diffuse into the deeper layers of the epidermis with D approximately 10(-17) m2/s, and that its elimination from the stratum corneum is mainly due to desquamation.

Electrochemical determination of femtomole amounts of free reduced and oxidized glutathione. Application to human hair follicles.

A simple and sensitive high-performance liquid chromatographic technique has been developed for the quantification of free reduced and free oxidized glutathione in biological samples. After acidic extraction and isocratic separation of the compounds of interest on a reversed-phase column, both forms of glutathione are quantified with a coulometric detector working in the oxidative mode. The limit of detection is 125 fmol for reduced glutathione and 400 fmol for the oxidized form (signal-to-noise ratio of 3). This sensitivity allows the measurement of the small amount of glutathione present in a single hair follicle. The technique is well adapted to microsamples, i.e. for non-invasive sampling technique (hair, skin, tears, etc.) and can be adapted to various cells or tissues.

Direct quantitative digital autoradiography-thin-layer chromatography of 3 alpha,3 beta- and 5 alpha-reduced and 17 beta-dehydrogenated androgens derived from testosterone metabolism.

A digital autoradiographic-thin-layer chromatographic method involving simple steps is described for thorough separation of eight major androgens (testosterone, androstenedione, dihydrotestosterone, androsterone, epiandrosterone, androstanedione, 3 alpha-androstanediol and 3 beta-androstanediol) derived from testosterone metabolism. Their direct quantification is performed by radioscanning, which avoids tedious plate-scraping and liquid scintillation counting. Satisfactory accuracy is obtained by both external standardization of plates by calibrated amounts of radiolabelled standard steroids and internal standardization of bioassays by radiolabelled squalane. Coefficients of variation are below 8.5% in the range 100-500 dpm. Some analytical criteria related to chromatographic conditions and quantification parameters depending on position-sensitive proportional counter are discussed.

In vivo percutaneous absorption: a key role for stratum corneum/vehicle partitioning.

Percutaneous absorption of five compounds was studied in the hairless rat in vivo: benzoic acid, caffeine, hydrocortisone, inulin and thiourea. The results clearly demonstrate that, as with in vitro experiments, a steady-state flux can be achieved in vivo. This steady-state flux is strongly molecule dependent. Thus, the values for inulin and benzoic acid differ by a factor of about 40. In contrast, although the physicochemical properties of the studied compounds vary widely, their lag times were not significantly different. The mean lag time was 11 +/- 2 min. Different compounds could be considered to have approximately the same apparent diffusion coefficient with regard to their percutaneous absorption in vivo. Thus, for a given thickness of stratum corneum and a given anatomical site, the penetration flux value of a substance depends only on its stratum corneum/vehicle partition coefficient. Using a classical model, we have demonstrated that the amount of substance present in the stratum corneum (Qsc) at equilibrium (30 min) is related to this partition coefficient. There is also a linear relationship between steady-state flux and Qsc. In practice, the in vivo steady-state flux of penetration of a compound can be predicted from the simple measurement of the amount present in the stratum corneum after a contact time of 30 min.

Study of the re-oiling process of hair with the replica technique.

Synopsis In the case of individuals with oily hair, the sebum excreted to the scalp surface spreads over the hair during the days following hair washing. The migration of sebum from the roots to the ends of hairs creates a gradient which may be measured by making casts on appropriate materials. An optical reading device enables the assessment of the alterations of the material brilliancy as a function of the sebum presence. The following parameters may be assessed from the recording: length of the cast, surface, height and width of the peak. Thus, different types of oily states may be identified according to: - the number of days after the last shampoo; - the characteristics of sebum (e.g. quantity, viscosity); - the features of hair. The casts reflect the oily state of hair as accurately as if it were assessed with a sensory method. Thus, this technique enables the study and comparison of oily states. It can be used to determine the efficacy of products having an effect on the re-oiling process of hair.