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Pigment cell refugia in homeotherms--the unique evolutionary position of the iris.

Homeotherms are generally considered to lack classical active dermal pigment cells (chromatophores) in their integument, attributable to the development of an outer covering coat of hair or feathers. However, bright colored dermal pigment cells, comparable to chromatophores of lower vertebrates, are found in the irides of many birds. We propose that, because of its exposed location, the iris is an area in which color from pigment cells has sustained a selective advantage and appears to have evolved independently of the general integument. In birds, the iris appears to have retained the potential for the complete expression of all dermal chromatophore types. Differences in cell morphology and the presence of unusual pigments in birds are suggested to be the result of evolutionary changes that followed the divergence of birds from reptiles. By comparison, mammals appear to have lost the potential for producing iridophores, xanthophores, or erythrophores comparable to those of lower vertebrates, even though some species possess brightly colored irides. It is proposed that at least one species of mammal (the domestic cat) has recruited a novel iridial reflecting pigment organelle originally developed in the choroidal tapetum lucidum. The potential presence of classical chromatophores in mammals remains open, as few species with bright irides have been examined.

Animals↗

Arbutin increases the pigmentation of cultured human melanocytes through mechanisms other than the induction of tyrosinase activity.

We assessed the effects of arbutin on the pigmentation of cultured normal human melanocytes. As indicated by a cell-blotting assay, arbutin at concentrations in the range of 0.5-8 mM increased the pigmentation of the cultured melanocytes, while kojic acid at concentrations in the range of 0.5-4 mM decreased the pigmentation. The pigmentation-augmenting effect of arbutin was further confirmed by the results of a cell-pelleting assay, the traditional method of assessment. Treatment of the cells with arbutin increased the melanin content of the cells and the protein content as well. On the other hand, the tyrosinase activity in the cells was reduced by arbutin treatment. The levels of transcription of tyrosinase and tyrosinase related protein-1 genes were not affected by arbutin treatment as indicated by a semi-quantitative reverse transcription-polymerase chain reaction assay. These results demonstrate that arbutin promotes an increase in pigmentation of cultured human melanocytes that is not mediated by augmented tyrosinase activity.

Arbutin↗

In vivo estimation of pigmentation in ultraviolet-exposed hairless mice.

A new in vivo method of visual scoring of pigmentation in hairless hr/hr mice with a C3H/Tif background is described. The mice were placed under a bank of 6 Philips TL08 fluorescent ultraviolet A (UVA) tubes in a dark room, and the pigmentation of the skin was compared with a Kodak Gray Scale with 20 different shades from white to black. The radiation from the tubes changed both the color of the back of the mouse and the gray scale into purple hues. The purple color of the back of each mouse could then be classified as one of 20 shades on the gray scale. An experiment was conducted exposing 3 groups of 20 mice to different doses of UV radiation from Philips TL01 tubes. One group of 20 mice was not irradiated and served as control. The pigmentation of each mouse was scored by one investigator every 2-3 weeks. After a few weeks of exposure a clear distinction between the groups was seen. To evaluate the inter- and intrapersonal variation of the method, 30 mice with various degrees of pigmentation were scored independently and blindly by two investigators. This was done twice during the study with a few days' interval. No interpersonal difference was found, but one investigator scored differently the first and second time by only 0.5 points. The described method provides a reproducible in vivo method, with very good discrimination, for estimation of pigmentation in hairless mice.

Animals↗

Photoprotection due to pigmentation and epidermal thickness after repeated exposure to ultraviolet light and psoralen plus ultraviolet A therapy.

Tanning and thickening of the epidermis are cardinal defensive responses of human skin to ultraviolet (UV) radiation that lead to increased photoprotection. Earlier studies have shown that skin pigmentation can be used to predict minimal erythema dose and minimal phototoxic dose. In this study it was calculated how much of the increase in photoprotection after 4 weeks of repeated exposure to suberythemogenic doses of either UVA or UVB radiation sources or psoralen plus UVA (PUVA) therapy that was attributable to melanogenesis. The backs of 12 volunteers were exposed to 6 different UVA and UVB radiation sources 9 times during 4 weeks. Skin pigmentation was assessed by skin reflectance measuring. Photoprotection was determined from the minimal erythema dose. Melanogenesis accounted for 63-95% of the increase in photoprotection after 4 weeks of exposure to UVA radiation. Exposure to two UVB sources induced a significant increase in photoprotection but not in pigmentation. Melanogenesis accounted only for 6-11% of the increase in photoprotection after 4 weeks of UVB exposure. The pigmentary and photoprotective responses to PUVA therapy were followed in 14 patients. After 2 weeks of exposure, the increase in photoprotection was significantly higher than predicted from the increase in skin pigmentation. After 4 weeks, melanogenesis accounted for only 36% of the increase in photoprotection. This study shows that melanogenesis accounts for the increased photoprotection after 2 weeks of exposure to UVA radiation, but after 4 weeks other protective mechanisms occur. During suberythemal UVB exposure and during PUVA therapy the importance of skin pigmentation in the overall photoprotection gradually decreases during a 4-week irradiation period.

Adult↗

Erythema and melanogenesis action spectra in heavily pigmented individuals as compared to fair-skinned Caucasians.

The protective role of epidermal melanin pigmentation against chronic exposure to ultraviolet radiation is widely accepted, although its photoprotective effect against acute exposure is less certain. In this study, the action spectra of erythema and melanogenesis in heavily pigmented individuals (skin type V) were determined at 295, 305, 315, and 365 nm, and compared with those of skin types I and II. When the erythema and melanogenesis action spectra for skin type V were normalized to 295 nm, they were identical to the corresponding action spectra for fair-skinned individuals, indicating that the photoprotection of epidermal melanin pigmentation is essentially independent of wavelength. The ratio of values for the minimum erythema dose (MED) between skin type V and skin types I and II was 2.29, which is close to the ratio of pigment in these skin types, as measured by diffuse reflectance spectroscopy in the visible range. The minimum immediate pigment darkening dose (IPD) and the minimum melanogenic dose (MMD) at 365 nm, and the MED and MMD at 315 nm were the same for all skin types, while the variation of MED for every skin type was maximum at 305 and 365 nm. The results provide circumstantial evidence that erythema and melanogenesis have the same mechanism at short-wavelength UVB (295 and 305 nm), and different mechanisms in UVA (365 nm). Furthermore, the 24 h MED at 305 nm appears to be a sensitive indicator of skin type.

Dose-Response Relationship, Radiation↗

Protection against minocycline pigment formation by ascorbic acid (vitamin C).

Minocycline, a member of the tetracycline family of antibiotics, is widely used in the treatment of acne. Its use has been associated with intrinsic staining of adult human teeth, bones, and soft tissues. It causes blackening of the thyroid glands in both animals and humans. It has been determined that the pigment is the product of an oxidation reaction. Laboratory studies have shown that the pigment formation can be induced by exposure to ultraviolet light in the presence of air, and that an antioxidant, such as ascorbic acid (vitamin C), can block its formation. The present study was undertaken to determine whether the antioxidant vitamin C could prevent the pigmentation of tissues of laboratory rats given minocycline. Based on other studies, one group of rats was given minocycline by stomach tube at a dose of 75 mg/kg/day, 5 days per week, for 6 weeks. A second experimental group was given the same regimen of minocycline plus a supplement of ascorbic acid at a level of 0.1% of the diet. A control group received no treatment at all; all rats were maintained on laboratory rats chow and water ad libitum. The rats were euthanized by carbon dioxide inhalation. Nasomaxillary bones, including central incisors, and thyroid glands were removed and fixed with 10% buffered formalin. Bones and teeth showed no gross signs of staining and, therefore, were not processed further. Thyroid glands were visibly darker in the minocycline group; specimens from all three groups were processed histologically. Microscopic examination revealed extensive deposits of black pigment throughout the follicles of the minocycline group, whereas the group receiving both minocycline and vitamin C showed no sign of pigmentation and were indistinguishable from controls. It is suggested that patients on long-term minocycline medication be monitored for thyroid function.

Animals↗

Rupture of retinal pigment epithelial detachment in senile macular disease.

Three eyes with senile detachment of the retinal pigment epithelium were complicated by tearing of the detached pigment epithelium with subsequent wrinkling or folding up of the pigment epithelium under the fovea. In all cases this only recently recognised complication of senile pigment epithelial detachment was documented before and after tearing by fluorescein angiography. In one eye pigment epithelial rupture was followed by rapid ingrowth of subretinal neovascularisation, and in one eye neovascularisation was suspected, although no definite new vessels could be observed.

Aged↗

Changes in the juxtapapillary retinal pigment epithelium following intravenous injection of sodium iodate. A light and electron microscopic study using horseradish peroxidase as a tracer.

The effect of an intravenous injection of sodium iodate on the retinal pigment epithelium immediately surrounding the optic nerve head has been investigated using horseradish peroxidase as a morphological tracer. A sodium iodate injection leads to a necrotic reaction of practically the complete retinal pigment epithelium. The juxtapapillary pigment epithelium, however, showed only attenuation and depigmentation without any necrotic reaction 4 and 12 days after the injection. Twenty-six days following the injection the epithelium had almost normalized. The result of this study thus demonstrates a noteworthy resistance of the juxtapapillary retinal pigment epithelium against the sodium iodate effect. At no stage was there a clear indication of a breakdown of the permeability barrier constituted by the juxtapapillary retinal pigment epithelium. Horseradish peroxidase moved into the peripapillary sensory retina by lateral diffusion from the surrounding retina and diffused into the optic nerve head proper, confirming the presence of a diffusional pathway through the Kuhnt intermediary tissue.

Animals↗

Absorption spectra of the hybrid pigments responsible for anomalous color vision.

Unequal homologous recombination events between green and red cone pigment genes produce the red-green or green-red hybrid pigment genes found in many individuals with variant color vision. Photobleaching difference absorption spectroscopy of hybrid pigments produced in cultured cells shows that the spectral sensitivity of each hybrid pigment is intermediate between the parental green and red pigment sensitivities. Amino acids encoded by exons 2, 3, 4, and 5 produce spectral shifts at the wavelength of maximal absorbance of 0 to 4, 0 to 4, 3 to 4, and 15 to 21 nanometers, respectively, the exact value depending on the identities of amino acids elsewhere in the hybrid.

Color Perception↗

Yolk pigments of the Mexican leaf frog.

Eggs of the Mexican leaf frog contain blue and yellow pigments identified as biliverdin and lutein, respectively. Both pigments are bound to proteins that occur in crystalline form in the yolk platelet. The major blue pigment is biliverdin IX alpha. The eggs vary in color from brilliant blue to pale yellow-green depending on the amount of each pigment. These pigments may provide protective coloration to the eggs.

Animals↗

New prodigiosin-like pigment from Alteromonas rubra.

The red prodigiosin-like pigment from Alteromonas rubra was shown to be a mixture of prodigiosin (pigment 1) and a new cyclic isomer (pigment 2). The new structure was elucidated by mass and nuclear magnetic resonance spectra. Careful examinations of the prodigiosins produced by Serratia marcescens, Vibrio psychoerythrus, and an unidentified red bacterium (LL-100-6) failed to disclose any of the new pigment, pigment 2.

Bacteria↗

Frequency of antibiotic and heavy metal resistance, pigmentation, and plasmids in bacteria of the marine air-water interface.

A field investigation of marine coastal waters revealed that the frequency of pigmented bacteria and the occurrence of bacterial antibiotic resistance were higher at the air-water interface than in the bulk water. The differences in the frequency of pigmented colonies at the surface and in the bulk-water samples could not be explained by the degree of cell surface hydrophobicity or by bacterial adhesion to air-water interfaces. Pigmented strains exhibited a higher degree of multiple drug resistance than did nonpigmented strains. However, the frequency of multiple drug resistance in nonpigmented strains was also substantial. An average of 91% of all strains were resistant to more than one antibiotic, and 21% of the bacteria isolated were resistant to five of the eight antibiotics tested. High numbers of plasmid-carrying strains were found among selected surface isolates, but the presence of detectable plasmids could not be correlated with either pigmentation or multiple drug resistance. Furthermore, selected surface isolates were significantly more resistant to mercury than were bulk-water bacteria. The higher frequency of pigmented, antibiotic-resistant, and mercury resistant strains at the air-water interface than in the bulk water are discussed in terms of various forms of selective pressure and genetic exchange at the surface.

Anti-Bacterial Agents↗

Extracellular polysaccharide from the black yeast NRRL Y-6272: improved methods for preparing a high-viscosity, pigment-free product.

When the extracellular polysaccharide from the black yeast NRRL Y-6272, composed of two parts N-acetyl-D-glucosamine and one part N-acetyl-D-glucosaminuronic acid, is isolated at maximum culture viscosity, adhering black pigment gives the polysaccharide preparations a gray-to-black appearance. Precipitation of the polysaccharide from cell-free culture supernatants with either ethanol of hexadecyltrimethylammonium bromide failed to remove the pigment. Various other methods were therefore tried for obtaining a high-viscosity polysaccharide product free of pigment. By systematically varying ingredients of defined and semidefined media, an improved medium was found that not only gave polysaccharide preparations of increased viscosity, but also increased yield. A key ingredient in this medium is L-asparagine. Also, adding autoclaved bovine serum albumin or egg albumin to this medium at the time of inoculation allowed a pigment-free polysaccharide to be isolated by standard procedures. None of several other proteins of synthetic polyamides tested were as effective as bovine serum albumin or egg albumin. In an alternate approach, pink mutants obtained by irradiation of the parent black strain with ultraviolet light, apparently produce the same extracellular polysaccharide free of any pigment but in lower yields or inferior in quality.

Acetylglucosamine↗

Influence of temperature of incubation and type of growth medium on pigmentation in Serratia marcescens.

Maximal amounts of prodigiosin were synthesized in either minimal or complete medium after incubation of cultures at 27 C for 7 days. Biosynthesis of prodigiosin began earlier and the range of temperature for formation was greater in complete medium. No prodigiosin was formed in either medium when cultures were incubated at 38 C; however, after a shift to 27 C, pigmentation ensued, provided the period of incubation at 38 C was not longer than 36 hr for minimal medium or 48 hr for complete medium. Washed, nonpigmented cells grown in either medium at 38 C for 72 hr could synthesize prodigiosin when suspended in saline at 27 C when casein hydrolysate was added. These suspensions produced less prodigiosin at a slower rate than did cultures growing in casein hydrolysate at 27 C without prior incubation at 38 C. Optimal concentration of casein hydrolysate for pigment formation by suspensions was 0.4%; optimal temperature was 27 C. Anaerobic incubation, shift back to 38 C, killing cells by heating, or chloramphenicol (25 mug/ml) inhibited pigmentation. Suspensions of washed cells forming pigment reached pH 8.0 to 8.3 rapidly and maintained this pH throughout incubation for 7 days. Measurements of viable count and of protein, plus other data, indicated that cellular multiplication did not occur in suspensions of washed cells during pigment formation. By this procedure utilizing a shift down in temperature, biosynthesis of prodigiosin by washed cells could be separated from multiplication of bacteria.

Anti-Bacterial Agents↗

Role of iron and sulfur in pigment and slime formation by Pseudomonas aeruginosa.

Media and an analytical scheme have been developed which allow both a qualitative and quantitative estimation of the formation of pyocyanine, related phenazines, pyorubrin, and a blue and a yellow-green fluorescent pigment by Pseudomonas aeruginosa. Use of the defined pyocyanine medium of Frank and DeMoss with sulfate or various organic sulfur sources allowed formation of pyocyanine, related phenazines, and pyorubrin. When sulfite was the sulfur source with or without iron, P. aeruginosa formed either a yellow-green or a blue fluorescent pigment. Formation of fluorescent pigments of P. aeruginosa is related to the ability of sulfite to act as a specific sulfur source. In an investigation of the role of both added iron and sulfur sources, complex patterns of pigment formation were observed. In addition to the fluorescent pigments, sulfite also supported the formation of slime by P. aeruginosa.

Culture Media↗

Biosynthesis of phenazine pigments in mutant and wild-type cultures of Pseudomonas aeruginosa.

Pigmentation mutants of Pseudomonas aeruginosa, selected by observed visual differences in coloration from the wild-type strain, were examined for altered patterns of phenazine synthesis. Three classes of mutants that were incapable of pyocyanine production were identified. Pigmentation patterns that were found to characterize the various mutant classes implicated precursor-product relationships, and a biochemical scheme covering the terminal reactions of pyocyanine biosynthesis is proposed. Among compounds tested as inhibitors of pigmentation, two effectively inhibited pyocyanine production production while allowing cell growth. p-Aminobenzoate inhibited total pigmentation; i.e., no other phenazine accumulated. m-Aminobenzoate inhibited a presumptive methylation step in pyocyanine biosynthesis, abolishing the formation of pyocyanine and aeruginosin pigments but increasing the yields of phenazine 1-carboxylic acid and oxychlororaphin. D-[2,3,4,5(n)-14C]shikimate was most efficiently incorporated into phenazines in the middle to late exponential phase of growth. Label was incorporated predominantly into pyocyanine in the absence of inhibitors and into phenazine 1-carboxylic acid when the organism was grown in the presence of m-aminobenzoate.

4-Aminobenzoic Acid↗

Production of actinorhodin-related "blue pigments" by Streptomyces coelicolor A3(2).

The genetically well-known strain Streptomyces coelicolor A3(2) produces the pH indicator (red/blue) antibiotic actinorhodin, but not all the "blue pigment" produced by this strain is actinorhodin. When the organism was subjected to various nutrient limitations (ammonium, nitrate, phosphate, or trace elements), and also during growth cessation caused by a relatively low medium pH, blue pigment production was initiated but the pigment and its location varied. At pH 4.5 to 5.5, significant formation of actinorhodin occurred and was located exclusively intracellularly. At pH 6.0 to 7.5 a different blue pigment was produced intracellularly as well as extracellularly. It was purified and identified as gamma-actinorhodin (the lactone form of actinorhodin). Analysis of act mutants of S. coelicolor A3(2) confirmed that both pigments are derived from the act biosynthetic pathway. Mutants with lesions in actII-ORF2, actII-ORF3, or actVA-ORF1, previously implicated or suggested to be involved in actinorhodin export, were impaired in production of gamma-actinorhodin, suggesting that synthesis of gamma-actinorhodin from actinorhodin is coupled to its export from the cell. However, effects on the level of actinorhodin production were also found in some mutants.

Anthraquinones↗

The 102-kilobase unstable region of Yersinia pestis comprises a high-pathogenicity island linked to a pigmentation segment which undergoes internal rearrangement.

Several pathogenicity islands have recently been identified in different bacterial species, including a high-pathogenicity island (HPI) in Yersinia enterocolitica 1B. In Y. pestis, a 102-kb chromosomal fragment (pgm locus) that carries genes involved in iron acquisition and colony pigmentation can be deleted en bloc. In this study, characterization and mapping of the 102-kb region of Y. pestis 6/69 were performed to determine if this unstable region is a pathogenicity island. We found that the 102-kb region of Y. pestis is composed of two clearly distinct regions: an approximately 35-kb iron acquisition segment, which is an HPI per se, linked to an approximately 68-kb pigmentation segment. This linkage was preserved in all of the Y. pestis strains studied. However, several nonpigmented Y. pestis strains harboring an irp2 gene have been previously identified, suggesting that the pigmentation segment is independently mobile. Comparison of the physical map of the 102-kb region of these strains with that of strain 6/69 and complementation experiments were carried out to determine the genetic basis of this phenomenon. We demonstrate that several different mechanisms involving mutations and various-size deletions are responsible for the nonpigmented phenotype in the nine strains studied. However, no deletion corresponded exactly to the pigmentation segment. The 102-kb region of Y. pestis is an evolutionarily stable linkage of an HPI with a pigmentation segment in a region of the chromosome prone to rearrangement in vitro.

Bacterial Outer Membrane Proteins↗