Search

The present study sought to determine whether the pigment produced by Proteus mirabilis from the L-forms of various aromatic amino acids under aerobic conditions is melanic in nature. It is a black-brown pigment which behaves like a melanin in many respects, namely solubility features, bleaching by oxidizing agents and positive response to the Fontana-Masson assay. In the present study, for the first time, it was shown by electron spin resonance analysis that a bacterial melanin is able to act as a free radical trap, as was previously demonstrated for other melanins. Scanning electron microscopy studies showed a specific organized structure of the pigment as rounded aggregates of spherical bodies. DNA hybridization data did not reveal, in the P. mirabilis genome, any nucleotide sequence related to Shewanella colwelliana mel A, one of the two melanogenesis systems already defined at the molecular level in bacteria. Results obtained from experiments on pigment production inhibition suggest a possible role of tyrosinase in P. mirabilis melanogenesis. In conclusion, from the bulk of our results, it appears that the pigment produced by P. mirabilis is melanic in nature.

The susceptibility to lipoperoxidation in liver of albino and pigmented Xenopus laevis Daudin, has been studied. Albino Xenopus liver was richer in polyunsaturated fatty acids (PUFAs) than the pigmented one; moreover, it was also richer in mitochondrial superoxide dismutase (MnSOD) and in reduced glutathione (GSH). The thiobarbituric acid-reactive substances (TBARS) values were more abundant in the albino tissue compared to the pigmented tissue both during spontaneous and Fe++ induced lipoperoxidation. Therefore, when isolated and purified melanin, in physiological quantities, was added to albino tissue, the TBARS values drastically decreased. Thus, melanin, in our experimental conditions, protects the albino tissue even more than SOD and GSH do. Melanin, in our opinion, acts as an antioxidant, because it is able to scavenge O2-.

The scavenger activity against O2-. of some flavonoids and melanins (synthetic melanins and melanins isolated from animal tissues, vegetable seeds, and mushroom spores) has been studied by ESR spectrometry. All these substances, except flavon and flavanone, diminish the signal of O2-. generated in vitro by a system containing H2O2 and acetone in an alkaline medium. It is shown that the presence of hydroxyl groups in the B ring of flavonoids is essential for their scavenger activity. Moreover, the presence of a hydroxyl at C-3 enhances the scavenger ability of flavonoids. Generally, aglycons are more active than their glycosides. It seems plausible that the antioxidant property of these substances comes from their scavenger activity against O2-(.). It is also pointed out that the scavenger activity shown by melanins, is strictly correlated with their nature of stable free radical.

The melanin content of Rana esculenta L. liver varies according to a circannual statistically significant rhythm, as shown by variance and single cosinor analysis. The maximum is found in autumn-winter, the minimum in spring-summer. The linear regression analysis shown a negative correlation between the amount of melanin and the environmental temperature.

The scavenger effect of melanin and of superoxide dismutase (SOD) activity on superoxide anion has been shown. In this work we show the relationship between melanin content and SOD activity in livers containing different quantities of melanin which were taken from various species of animals. The mitochondrial SOD activity disappears when the melanin content in the liver is very high; moreover it increases, in the liver of various species of animals examined, proportionally to the decrease of melanin content. No significant variation of the SOD activity localized in the soluble fraction has been detected when related to the melanin content. We think that in the pigmented liver the antioxidant activity of the melanin could mimic part of the function of SOD. The loss of Mn SOD activity could be mediated by a low intracellular level of superoxide anion due to the scavenger effect of melanin on superoxide anion; in fact, it is well known that the biosynthesis of Mn SOD is induced by intracellular levels of superoxide anion.

Statistical examination of the data from experiments conducted on groups of Amphibians induces us to retain as plausible the hypothesis which indicates in fasting one of the preminent factors that cause an increase in the melanic content of their liver. The temperature does not act directly, but indirectly, on the process of melanosynthesis, by inducing or otherwise the semilethargous state on which alimentation depends.

Frogs caught in winter and kept at summer temperatures, show an increase in the melanic content of the liver. Frogs caught in summer and kept at winter temperatures do not show variations in hepatic melanins with respect to the summer control specimes. This data does not correspond with the natural course of hepatic melanin fluctuations, therefore the causes of the variations are to be sought in factors other than the temperature, one of which could very probably be represented by alimentation.