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Effects of paprika pigments on oxidation of linoleic acid stored in the dark or exposed to light.

We examined the antioxidant effects of paprika pigments on oxidation of linoleic acid and on decoloration of the sample when stored at 37 degrees C in the dark or exposed to fluorescent light for 8 h per day. (1)H nuclear magnetic resonance with dioxane as an external proton reference was used to estimate the oxidative deterioration of linoleic acid. Oxidation was estimated by observing the ratio of the divinylmethylene proton signal area in linoleic acid vs the proton signal area in dioxane. The addition of paprika pigments suppressed the oxidation of linoleic acid during storage in the dark, and the effect was markedly increased with increasing concentrations (0.02, 0.2, and 2%). When the linoleic acid with added paprika pigments was exposed to light, only a slight suppression of oxidation was observed, and the color of the sample disappeared more rapidly than that in the dark. At the time of decoloration of the sample with added pigments, considerable oxidation of linoleic acid occurred. As the color change is due to degradation of the pigment, an increase in oxidation at the time of discoloration is consistent with the pigments functioning as antioxidants. The addition of alpha-tocopherol to paprika pigments stabilized degradation of the pigments by light. Although the addition of alpha-tocopherol to linoleic acid with added paprika pigments prolonged the decoloration of the sample under light, the prevention of oxidation under the light condition was not as effective as for the samples stored in the dark.

Antioxidants↗

Involvement of photooxidation of melanogenic precursors in prolonged pigmentation induced by ultraviolet A.

Melanin is usually produced in melanocytes; however, our histologic research indicates that brownish black persistent pigmentation occurs at the basal layer and, partially, at the suprabasal layer of the epidermis, outside of melanocytes, in skin exposed to high doses of ultraviolet (UV) A radiation. This pigmentation remains for several weeks. We examined whether colorless melanogenic precursors participate in this UVA-induced persistent pigmentation. Among them, 5,6-dihydroxyindole-2-carboxylic acid (DHICA) and its O-methyl metabolite, 6-hydroxy-5-methoxyindole-2-carboxylic acid (6H5MICA), can change into dark-colored pigment upon exposure to UVA, but not UVB, radiation, producing irreversible brownish black pigmentation. These hydroxyindole derivatives readily changed into dark-colored pigment upon UVA irradiation in the presence of hydrogen peroxide in vitro. Histochemical and biochemical studies demonstrated that 6H5MICA accumulates in excised skin specimens and supernatant of cultured human melanocytes. These results suggest that dark-colored pigment is generated directly, outside of melanocytes, in response to UVA exposure, by photooxidation of O-methyl metabolites of DHICA, such as 6H5MICA, at least. We propose that the pigmentation derived from this compound at the basal and suprabasal layers of the epidermis is involved in the mechanism of skin persistent pigmentation without reddening, observed after exposure to sunlight.

Adult↗

Gallbladder motility and cholesterol crystallization in bile from patients with pigment and cholesterol gallstones.

BACKGROUND: Little is known about gallbladder motility in patients with black pigment stones when compared to cholesterol gallstone patients, or about their relationship to biliary composition, crystallization and stone characteristics. DESIGN: Fasting and postprandial gallbladder volumes were studied by ultrasonography in 49 gallstone patients with pigment (n = 14) or cholesterol (n = 35) stones and 30 healthy controls. After cholecystectomy stone composition, gallbladder wall inflammation, cholesterol saturation index and appearance of platelike cholesterol crystals in bile were evaluated in gallstone patients. RESULTS: Fasting gallbladder volume was significantly (P < 0.05) increased in cholesterol stone patients (31.7 +/- 1.9 mL) but not in pigment stone patients (21.9 +/- 3.1 mL), compared to controls (21.0 +/- 1.5 mL). Postprandial emptying was delayed in patients (half-emptying time: 31 +/- 2 min, 35 +/- 3 min, 24 +/- 2 min in cholesterol stone patients, pigment stone patients and controls, respectively, P < 0.05) and incomplete (residual volume: 43.2 +/- 2.7%, 40.0 +/- 4.3%, 15.8 +/- 1.6% min in cholesterol stone patients, pigment stone patients and controls, respectively, P < 0.05). The inflammation of the gallbladder wall was mild or absent in all cases. Biliary cholesterol saturation index was 152.3 +/- 8.5% and 92.9 +/- 4.8% in patients with cholesterol and pigment stones, respectively (P < 0.01). Whereas cholesterol crystals never appeared during 21 days in biles from patients with pigment stones, crystal observation time in patients with cholesterol gallstone was 5 days (median) and was significantly shorter in patients with multiple (4 days) than in patients with solitary (12 days) cholesterol stones (P = 0.0019). CONCLUSIONS: Patients with black pigment stones who do not have excess cholesterol and do not grow cholesterol crystals in bile have decreased gallbladder emptying, although to a lesser extent than patients with cholesterol stones. Thus, gallbladder stasis is likely to put a subset of subjects at risk for the formation of pigment gallstones, and pathogenic mechanisms need to be further investigated.

Adult↗

Further characterization of pigment-producing Malassezia strains.

Reference as well as field strains of Malassezia furfur (30), M. sympodialis (49), M. globosa (52), M. obtusa (one), M. restricta (one), M. slooffiae (seven), and M. pachydermatis (373) were investigated for their ability to produce pigment and fluorochromes when tryptophan (Trp) is offered as the main nitrogen source. Only the M. furfur strains produced pigment on a pigment-inducing medium (p-medium). Remarkably, the optical activity of Trp was not significant for pigment synthesis. Other nitrogen sources that are structurally similar to Trp (gramine, tryptamine, serotonin) did not induce pigment formation. All lipophilic non-furfur species failed to grow and to form pigment on this agar. However, growth of all lipid-dependent species was achieved on a modified Dixon agar in which peptone had been substituted by an equal amount of l-Trp. Here, too, the M. furfur colonies were characterized by rapidly developing dark brown halos. Furthermore, about 11% of the M. pachydermatis strains tested produced pigment formation on the p-medium, which was enhanced by addition of d-glucose. In contrast to M. furfur, pigment formation occurred after a markedly longer incubation time (4 weeks unlike 3-5 days) with a lower yield and limited color spectrum (thin layer chromatography, TLC). The UV filter pityriacitrine recently described for M. furfur was also demonstrated for M. pachydermatis by extraction, high-performance liquid chromatography (HPLC) analysis with co-elution and mass spectroscopy. The phenotypic feature of pigment formation in some strains of M. pachydermatis may confirm recent molecular-genetic findings suggesting a relationship between some strains of M. pachydermatis and M. furfur.

Alkaloids↗

Establishment of pigment cell lineage in embryos of the sea urchin, Hemicentrotus pulcherrimus.

In an attempt to estimate the number of pigment precursor cells in sea urchin embryos, DNA synthesis and cell divisions were blocked with aphidicolin from various stages of development. Interestingly, pigment cells differentiated on a normal time schedule, even if the embryos were treated from late cleavage stages on. In most of the embryos treated from 10 h on, 10-15 pigment cells differentiated. Thereafter, the number of pigment cells in the aphidicolin-treated embryos further increased, as the initiation of the treatment was delayed. On the other hand, total cell volumes in the pigment lineage, calculated from the averaged number and diameter of differentiated pigment cells, were almost the same irrespective of the time of the initiation of aphidicolin treatment. This indicated that the increase in the number was caused by divisions of the pre-existing cells in the pigment lineage. Thus, the founder cells that exclusively produce pigment cells could be identified. They are nine times-cleaved blastomeres and specified by 10 h post-fertilization. The obtained results also clarified the division schedule in the pigment lineage; the founder cells divide once (10th) until hatching, and divide once more (11th) by the end of gastrulation.

Animals↗

Production of red pigment by submerged culture of Paecilomyces sinclairii.

AIMS: From a survey of submerged culture of edible mushrooms, a high pigment-producing fungus Paecilomyces sinclairii was selected and its optimal culture conditions investigated. METHODS AND RESULTS: The optimal culture conditions for pigment production were as follows: inoculum age, 3 d; temperature, 25 degrees C; initial pH, 6.0; carbon source, 1.5% (w/v) soluble starch; nitrogen source, 1.5% (w/v) meat peptone. Although addition of 10 mmol l(-1) CaCl2 to the culture medium slightly increased pigment production, most of the bio-elements examined had no notable or detrimental effect on pigment production. CONCLUSIONS: Under the optimal conditions obtained in the flask culture tested, a ninefold increase in pigment production (4.4 g l(-1)) was achieved using a 5(-l) batch fermenter. Paecilomyces sinclairii secreted water-soluble red pigment into the culture medium. The pigment colour was strongly dependent on the pH of the solution: red at pH 3-4, violet at pH 5-9 and pink at pH 10-12. SIGNIFICANCE AND IMPACT OF THE STUDY: The high concentration of pigment (4.4 g l(-1)) produced by P. sinclairii demonstrates the possibility of commercial production of pigment by this strain, considering its relatively high production yield and light stability.

Bioreactors↗

Drosophila pigmentation evolution: divergent genotypes underlying convergent phenotypes.

Similar phenotypic changes have evolved independently in many animal taxa. It is unknown whether independent changes involve the same or different developmental and genetic mechanisms. Myriad pigment patterns in the genus Drosophila offer numerous opportunities to address this question. Previous studies identified regulatory and structural genes involved in the development and diversification of pigmentation in selected species. Here, we examine Drosophila americana and Drosophila novamexicana, interfertile species that have evolved dramatic pigmentation differences during the few million years since their divergence. Interspecific genetic analysis was used to investigate the contribution of five specific candidate genes and other genomic regions to phenotypic divergence by testing for associations between molecular markers and pigmentation. At least four distinct genomic regions contributed to pigmentation differences, one of which included the ebony gene. Ebony protein was expressed at higher levels in the more yellow D. novamexicana than the heavily melanized D. americana. Because Ebony promotes yellow pigment formation and suppresses melanization, the expression difference and genetic association suggest that evolution at the ebony locus contributed to pigmentation divergence between D. americana and D. novamexicana. Surprisingly, no genetic association with the yellow locus was detected in this study, and Yellow expression was identical in the two species. Evolution at the yellow locus underlies pigmentation divergence among other Drosophila species; thus, similar pigment patterns have evolved through regulatory changes in different genes in different lineages. These findings bear upon understanding classic models of melanism and mimicry.

Animals↗

Sequence divergence of the red and green visual pigments in great apes and humans.

We have determined the coding sequences of red and green visual pigment genes of the chimpanzee, gorilla, and orangutan. The deduced amino acid sequences of these pigments are highly homologous to the equivalent human pigments. None of the amino acid differences occurred at sites that were previously shown to influence pigment absorption characteristics. Therefore, we predict the spectra of red and green pigments of the apes to have wavelengths of maximum absorption that differ by < 2 nm from the equivalent human pigments and that color vision in these nonhuman primates will be very similar, if not identical, to that in humans. A total of 14 within-species polymorphisms (6 involving silent substitutions) were observed in the coding sequences of the red and green pigment genes of the great apes. Remarkably, the polymorphisms at 6 of these sites had been observed in human populations, suggesting that they predated the evolution of higher primates. Alleles at polymorphic sites were often shared between the red and green pigment genes. The average synonymous rate of divergence of red from green sequences was approximately 1/10th that estimated for other proteins of higher primates, indicating the involvement of gene conversion in generating these polymorphisms. The high degree of homology and juxtaposition of these two genes on the X chromosome has promoted unequal recombination and/or gene conversion that led to sequence homogenization. However, natural selection operated to maintain the degree of separation in peak absorbance between the red and green pigments that resulted in optimal chromatic discrimination. This represents a unique case of molecular coevolution between two homologous genes that functionally interact at the behavioral level.

Amino Acid Sequence↗

Mechanisms of spectral tuning in the mouse green cone pigment.

Diversification of cone pigment spectral sensitivities during evolution is a prerequisite for the development of color vision. Previous studies have identified two naturally occurring mechanisms that produce variation among vertebrate pigments by red-shifting visual pigment absorbance: addition of hydroxyl groups to the putative chromophore binding pocket and binding of chloride to a putative extracellular loop. In this paper we describe the use of two blue-shifting mechanisms during the evolution of rodent long-wave cone pigments. The mouse green pigment belongs to the long-wave subfamily of cone pigments, but its absorption maximum is 508 nm, similar to that of the rhodopsin subfamily of visual pigments, but blue-shifted 44 nm relative to the human red pigment, its closest homologue. We show that acquisition of a hydroxyl group near the retinylidene Schiff base and loss of the chloride binding site mentioned above fully account for the observed blue shift. These data indicate that the chloride binding site is not a universal attribute of long-wave cone pigments as generally supposed, and that, depending upon location, hydroxyl groups can alter the environment of the chromophore to produce either red or blue shifts.

Amino Acid Sequence↗

Identification of transplanted retinal pigment epithelium with a novel chromosomal marker.

PURPOSE: To demonstrate the ability of a novel chromosomal marker to identify retinal pigment epithelium (RPE) after xenotransplantation, and determine the short-term correlation between pigment and this nuclear marker. METHODS: Primary pigmented RPE harvested from third trimester fetal pigs were transplanted as microaggregates into the subretinal space of 3 albino rabbits. We then used an in situ probe for a repetitive segment of the porcine chromosome to identify the transplanted RPE. RESULTS: Pigmented cells were visible in the subretinal space 2 weeks after transplantation. Approximately 70% of pigment-containing cells were also labeled with the porcine chromosomal marker. Labeled cells were predominantly flatter in morphology and close to Bruch's membrane whereas unlabeled cells were rounder and further from Bruch's membrane. The outer nuclear layer thickness was normal above the pigmented monolayer but was decreased over areas containing multiple layers of pigmented cells. CONCLUSIONS: Fetal porcine RPE xenografts can be identified with a nuclear marker for a repetitive segment of the porcine chromosome. The presence of pigment within unlabelled cells suggests that pigment is not a robust marker for transplanted RPE.

Animals↗

The development of pigment granules in the eyes of wild type and mutant Drosophila melanogaster.

The eye pigment system in Drosophila melanogaster has been studied with the electron microscope. Details in the development of pigment granules in wild type flies and in three eye color mutants are described. Four different types of pigment granules have been found. Type I granules, which carry ommochrome pigment and occur in both primary and secondary pigment cells of ommatidia, are believed to develop as vesicular secretions by way of the Golgi apparatus. The formation of Type II granules, which are restricted to the secondary pigment cells and contain drosopterin pigments, involves accumulation of 60- to 80-A fibers producing an elliptical granule. Type III granules appear to be empty vesicles, except for small marginal areas of dense material; they are thought to be abnormal entities containing ommochrome pigment. Type IV granules are characteristic of colorless mutants regardless of genotype, and during the course of development they often contain glycogen, ribosomes, and show acid phosphatase activity; for these reasons and because of their bizarre and variable morphology, they are considered to be autophagic vacuoles. The 300-A particles commonly found in pigment cells are identified as glycogen on the basis of their morphology and their sensitivity to salivary digestion.

Acid Phosphatase↗

Multiple visual pigments in a photoreceptor of the salamander retina.

Although a given retina typically contains several visual pigments, each formed from a retinal chromophore bound to a specific opsin protein, single photoreceptor cells have been thought to express only one type of opsin. This design maximizes a cell's sensitivity to a particular wavelength band and facilitates wavelength discrimination in retinas that process color. We report electrophysiological evidence that the ultraviolet-sensitive cone of salamander violates this rule. This cell contains three different functional opsins. The three opsins could combine with the two different chromophores present in salamander retina to form six visual pigments. Whereas rods and other cones of salamander use both chromophores, they appear to express only one type of opsin per cell. In visual pigment absorption spectra, the bandwidth at half-maximal sensitivity increases as the pigment's wavelength maximum decreases. However, the bandwidth of the UV-absorbing pigment deviates from this trend; it is narrow like that of a red-absorbing pigment. In addition, the UV-absorbing pigment has a high apparent photosensitivity when compared with that of red- and blue-absorbing pigments and rhodopsin. These properties suggest that the mechanisms responsible for spectrally tuning visual pigments separate two absorption bands as the wavelength of maximal sensitivity shifts from UV to long wavelengths.

Animals↗

A study on the removal of the melanin pigmentation of dog gingiva by CO 2 laser irradiation.

Removal of melanin pigmentation of the gingiva with CO 2 laser irradiation was investigated. The melanin-pigmented gingiva of 5 dogs were classified according to the degree of gingival pigmentation (slight, moderate, severe) and its extent (local, extensive). CO 2 laser was irradiated on the surface of the pigmented gingiva under the following conditions; defocused beams of spot size 4 mm, irradiation time 0.2(S), output 6-8(W)). That is, the irradiation energy density was 9.6-12.8 J/cm 2. The irradiated gingiva was examined macroscopically and histopathologically. The histopathological changes were investigated by hematoxylin-eosin staining and Masson staining for the differential diagnosis of melanin pigmentation cells. These examinations were done one week and three weeks after the laser irradiation. Immediately after irradiation, the gingiva surface showed white color and bulla-like appearance. One week after irradiation, the pigmentation of the lased gingiva disappeared in all cases macroscopically. Histologically, the pigmented-laden cells could not be found and no inflammatory cell infiltration was observed. Three weeks after irradiation, no reappearance of pigmentation was observed in either macroscopical or microscopical investigations excluding severe/extensive cases. These findings suggested that it is possible to remove gingival melanin pigmentation by CO 2 laser irradiation.

Animals↗

Formation and role of malaria pigment.

Malaria pigment is most abundant and distinct in gametocytes. Trophozoites have variable amounts of pigment, depending on the species of Plasmodium and the stage of infection. In Plasmodium falciparum infection, blood smear preparations fall into two categories that are distinguishable at all levels of parasitemia; one type of preparation contains only pigment-deficient trophozoites, and the other type contains only pigment-rich trophozoites. Pigment accumulates in the residual body of the mature schizont and is lost upon rupture of the schizont. In contrast, pigment remains associated with the macrogametocyte and developing oocyst. Certain antimalarial drugs, such as chloroquine, have distinct effects on pigment clumping. These observations raise questions regarding the current idea of pigment as an inert excretory product of hemoglobin metabolism. It is suggested that pigment particles represent stacked utilizable intermediates of hemoglobin digestion that accumulate in the gametocyte to serve as a food reserve during the growth cycle in the mosquito.

Animals↗

Detection of a novel pigment network feature in reticulated black solar lentigo by high-resolution epiluminescence microscopy.

Epiluminescence light microscopy (ELM) of pigmented skin lesions has led to a catalog of pigment network (PN) features. The objective of this study was to determine whether high-resolution ELM detects additional pigment structures not seen with conventional ELM. Epiluminescence light microscopy was performed by placing the lens of a standard light microscope directly on the skin surface, with resulting enhanced optical resolution compared with ELM systems currently in general use. Eight reticulated black solar lentigines were studied. All lesions were viewed and analyzed using dermatoscopic criteria for the PN. In addition, they were all photographed, excised, and examined histologically. Two subtypes of black solar lentigines could be distinguished using generally accepted dermatoscopic criteria for the PN. Furthermore, a new pigment structure was detected, namely, pigment spots of equal color, shape, and size, which were regularly superimposed on and juxtaposed to the PN. Clinicopathologic correlation showed these spots to represent individual hyperpigmented corneocytes. Because such cells result from physiologic excessive pigment translocation via the epidermal melanocytic unit, we called them pigmented corneocytes. Pigmented corneocytes were seen in seven of eight black solar lentigines. The ELM technique presented here allows for more detailed analysis and classification of the PN components. Pigmented corneocytes are proposed as an additional dermatoscopic criterion.

Adult↗

Lipofuscin pigmentation (so-called "melanosis") of the prostate.

Although intraepithelial pigment in the prostate gland has been termed melanosis, the nature of the pigment is not entirely clear, and many pathologists are not aware of its existence. We examined 863 hematoxylin and eosin (H + E) stained slides from 150 surgical specimens of prostate (69 needle biopsies, 66 transurethral resections, 14 radical prostatectomies, and 1 suprapubic prostatectomy) from 149 patients (age range, 47 to 90 years; mean 70 years) in an effort to characterize this pigment. The 1-3 microns in diameter, predominantly subnuclear, yellow-brown to gray-brown granules with a dark blue rim (by H + E) stained positively with Fontana-Masson, periodic acid-Schiff with diastase, Congo red, luxol fast blue, and oil-red-O and exhibited yellow autofluorescence consistent with lipofuscin. H + E stained slides revealed pigment in the benign epithelium in 86 of 150 cases (57%), within stromal macrophages in eight cases, and in atypical epithelium in two cases of high-grade prostatic intraepithelial neoplasia. Ten cases of invasive adenocarcinoma without recognizable pigment in H + E stained sections were stained by the Fontana-Masson technique, and pigment was identified in malignant epithelium in three of these cases. Ultrastructural examination of intraepithelial pigment in KII-fixed tissue from three radical prostatectomy specimens demonstrated the typical appearance of lipofuscin. Although intraepithelial pigment in prostatic biopsy or resection specimens is usually considered characteristic of seminal vesicle epithelium, our study demonstrates that lipofuscin is commonly present in epithelial cells of benign prostatic hyperplasia and less frequently in those of prostatic intraepithelial neoplasia and adenocarcinoma. The recognition of this pigment is important in preventing diagnostic confusion with seminal vesicle epithelium and with melanocytic lesions.

Aged↗

Evolution of abdominal pigmentation differences across species in the Drosophila dunni subgroup.

The Drosophila dunni subgroup displays a nearly perfect latitudinal cline in abdominal pigmentation that likely resulted from selective forces acting in the habitat of each species during speciation. Here we characterize the nature of this clinal variation by developing a quantitative measure to assess variation in abdominal pigmentation within and between the D. dunni subgroup species. Using discriminant analysis, we confirm the existence of a cline and find that our quantitative measure of pigmentation distinguishes each of the species with singular efficacy. We then combine our quantitative phenotypic analysis of pigmentation with the phylogeny of the D. dunni subgroup species and map the species relationships into the three-dimensional morphological space defined by our pigmentation measures. In this manner, we can visualize how the species have traversed the morphological pigmentation space during the course of speciation. Our analysis reveals that natural selection has caused overall intensity of pigmentation among the northernmost species of the cline to converge. Along with this convergence in phenotype has been a relaxation in expression of sexual dimorphism in these species, indicating a possible shift in the relative intensity of natural and sexual selection. Our analysis indicates an accelerated rate of change in pigmentation for the darkest species in addition to this species evolving a novel abdominal pigmentation trait.

Abdomen↗

Genetics of a difference in pigmentation between Drosophila yakuba and Drosophila santomea.

Drosophila yakuba is a species widespread in Africa, whereas D. santomea, its newly discovered sister species, is endemic to the volcanic island of São Tomé in the Gulf of Guinea. Drosophila santomea probably formed after colonization of the island by its common ancestor with D. yakuba. The two species differ strikingly in pigmentation: D. santomea, unlike the other eight species in the D. melanogaster subgroup, almost completely lacks dark abdominal pigmentation. D. yakuba shows the sexually dimorphic pigmentation typical of the group: both sexes have melanic patterns on the abdomen, but males are much darker than females. A genetic analysis of this species difference using morphological markers shows that the X chromosome accounts for nearly 90% of the species difference in the area of abdomen that is pigmented and that at least three genes (one on each major chromosome) are involved in each sex. The order of chromosome effects on pigmentation area are the same in males and females, suggesting that loss of pigmentation in D. santomea may have involved the same genes in both sexes. Further genetic analysis of the interspecific difference between males in pigmentation area and intensity using molecular markers shows that at least five genes are responsible, with no single locus having an overwhelming effect on the trait. The species difference is thus oligogenic or polygenic. Different chromosomal regions from each of the two species influenced pigmentation in the same direction, suggesting that the species difference (at least in males) is due to natural or sexual selection and not genetic drift. Measurements of sexual isolation between the species in both light and dark conditions show no difference, suggesting that the pigmentation difference is not an important cue for interspecific mate discrimination. Using DNA sequence differences in nine noncoding regions, we estimate that D. santomea and D. yakuba diverged about 400,000 years ago, a time similar to the divergences between two other well-studied pair of species in the subgroup, both of which also involved island colonization.

Animals↗