Melanin density and melanin type predict melanocytic naevi in 19-20 year olds of northern European ancestry.

Recent advances in estimating the density of cutaneous melanin by spectrophotometry and the concentration of the two types of melanin (eumelanin and phaeomelanin) in hair offer the potential to define the risk of skin cancer in individuals more accurately. The presence of common melanocytic naevi on the arm is associated with an increased risk of melanoma, and in this study the associations of arm naevi with melanin density at the upper inner arm and with melanin type in hair samples were examined in a representative sample (n = 267) of 19-20 year olds of northern European ancestry. Particularly in men, the association with naevus count was stronger for cutaneous melanin density than for follicular melanin type. Adjusted for recreational sun exposure, the rank correlation coefficients were r = -0.25, 0.12 and 0.01 for men, and r = -0.17, -0.12 and 0.14 for women, for cutaneous melanin, hair eumelanin and hair phaeomelanin, respectively. The associations with less objective markers of phenotype (hair colour, eye colour, nurse-assessed skin colour, and self-reported skin reaction to unaccustomed sun) were weaker. These findings provide important new information that human susceptibility to mutations of melanocytes can be estimated by objective biological measures. The next step is to determine whether these measures also predict the risk of melanoma.

Latanoprost stimulates eumelanogenesis in iridial melanocytes of cynomolgus monkeys.

Latanoprost, the active principle of Xalatan eye drops, is a prostaglandin F2alpha analogue in widespread use for the treatment of glaucoma. During chronic treatment with the drug, an increased pigmentation of the iris was observed in both primates and man. To gain an insight into the nature of this effect, we analyzed the stroma of the irides of cynomolgus monkeys subjected to 25-38 weeks of treatment. A highly sensitive procedure, based on chemical degradation by alkaline hydrogen peroxide oxidation, or hydriodic acid hydrolysis, was developed, which allowed eumelanin and pheomelanin analysis of a single iris at a time. Untreated monkey irides were found to be essentially pheomelanic, providing further support to the recently reported occurrence of these pigments in human irides. In the Latanoprost-treated eyes, the amount of eumelanin increased from three to sevenfold, while the variation of pheomelanin did not exceed 25%. The increase in eumelanin/pheomelanin ratio in the treated eyes, as compared with the contralateral control eyes, varied from three to fivefold, and the change was statistically significant (P < 0.01; t-test). Based on the results of parallel studies, showing that Latanoprost does not induce proliferation of iridial melanocytes, and that the other pigmented layers of the iris which do not contain melanocytes are not affected by the drug, it can be concluded that the observed effect is a result of a direct interaction with the melanogenic mechanism. This probably involves activation of tyrosinase, as suggested, to account for the stimulation of melanin synthesis by related compounds, including natural prostaglandins.

Microanalysis of melanins in mammalian hair by alkaline hydrogen peroxide degradation: identification of a new structural marker of pheomelanins.

A highly sensitive, easy-to-perform method for melanin analysis in pigmented tissues based on alkaline hydrogen peroxide degradation has been developed and accomplishes simultaneous determination of eumelanins and pheomelanins. Pyrrole-2,3,5- tricarboxylic acid, the typical eumelanin marker, was obtained in higher yields than in previous procedures. A benzothiazole acid, 6-(2-amino-2-carboxyethyl)-2-carboxy-4-hydroxybenzothiazole, characterized in our previous studies as a specific marker of pheomelanins, and the newly identified 1,3-thiazole-2,4, 5-tricarboxylic acid were also used for pigment analysis. Optimal yields of the pigment markers were obtained at 24 h reaction time. Pyrrole-2,3,5-tricarboxylic acid, 6-(2-amino-2-carboxyethyl)-2-carboxy-4-hydroxybenzothiazole, and 1, 3-thiazole-2,4,5- tricarboxylic acid were quantified in a single chromatographic analysis without fractionation or work up of the degradation mixture. The linearity (linearity coefficient from 0.997 to 0.999) was excellent and the inter-assay (percentage coefficient of variation values in the range 0.2-2, n = 6) and intra-assay (percentage coefficient of variation values

Characterization of melanins in human irides and cultured uveal melanocytes from eyes of different colors.

The presence of eumelanin and pheomelanin in irides from eyes of various colors was determined and quantified by a highly specific microanalytical procedure based on chemical degradation. Significant differences in the type of melanin were observed in the stroma and iris pigment epithelium (IPE) fractions obtained by micro-dissection of the iris specimens. Melanin from the IPE is essentially eumelanin, while the pigment in IPE-scraped iris (consisting mainly of stroma plus anterior IPE) proved to be both eumelanic and pheomelanic. A pheomelanic-type pigmentation was associated with green irides, while green-blue mixed-color irides were mostly eumelanic; by contrast, green-brown mixed-color and brown irides could not be placed into either of the two categories and probably feature a mixed pigment content. Blue irides invariably exhibited very low pigment content. Analysis of cultured iridial melanocytes in the growing stage showed a significant shift to pheomelanic pigmentation when compared with those in IPE-scraped tissues, providing evidence that growth of iridial melanocyte induce a marked change of melanin metabolism. After senescence, cultured melanocytes exhibited a marked increase in pigment content, most of the variation was associated with the eumelanin content. These results represent the first direct evidence for the presence of eumelanin and pheomelanin in human irides, and suggest that differences in stromal pigmentation are due not only to the quantity, but also the nature of the melanin pigment.

Phaeomelanin versus eumelanin as a chemical indicator of ultraviolet sensitivity in fair-skinned subjects at high risk for melanoma: a pilot study.

It is now generally agreed that solar exposure is a major external factor in the causation of cutaneous melanoma in light skinned populations with red hair and a marked susceptibility to the acute effects of ultraviolet (UV) radiation. In the present study, we investigated the existence of a possible relationship between hair melanin composition and minimal erythema dose (MED), as an indicator of UV sensitivity, in a group of 15 healthy red-haired subjects aged 20-46 years. In spite of comparable skin and hair colour, marked variations were observed in the MED values as well as in the hair melanin composition. Phaeomelanin levels varied in the range 0.026-0.53% w/w and were generally comparable to or higher than eumelanin levels (0.042-0.17% w/w). No significant relationship was found between MED values and phaeomelanin, eumelanin or total melanin (eumelanin plus phaeomelanin) content. Notably, however, a gross positive correlation was found between the eumelanin/phaeomelanin ratio and the MED values. These results would suggest that a high UV sensitivity is associated with high phaeomelanin and low eumelanin levels, and point to the eumelanin/phaeomelanin ratio as a novel chemical parameter that could be used for predicting individuals at high risk for skin cancer and melanoma.

Comparative analysis of melanins and melanosomes produced by various coat color mutants.

Pigmentation in the skin, hair, and eyes of animals is influenced by a number of genes that modulate the activity of melanocytes, the intervention of enzymatic controls at different stages of the melanogenic process, and the physico-chemical properties of the final pigment. The results of combined phenotypic, ultrastructural, biochemical, and chemical analyses of hairs of a variety of defined genotypes on a common genetic background performed in this study are consistent with the view that pigmentation of dark to black hairs results from the incorporation of eumelanin pigments whereas that of yellow hairs results from the incorporation of eu- and pheomelanins. It is also clear that relatively minor differences in melanin content can have dramatic effects on visible hair color. A good correlation was found for expression of (and enzyme activities associated with) TRP1 and TRP2 with eumelanin synthesis and eumelanosome production.