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Characterization of the yellow-pigment genes of Erwinia herbicola.

A 6.7 kb DNA fragment containing the pigment genes of Erwinia herbicola Eho13 has been cloned into Escherichia coli. These genes were chromosomally encoded in E. herbicola. The entire DNA fragment could be divided into at least three regions. Deletions in Region I resulted in a non-pigmented phenotype, a deletion in Region II resulted in a pink/yellow phenotype, deletions in Region III resulted in either a pink or a non-pigmented phenotype. Tn1000 insertions in the same regions, however, gave different phenotypes. Insertions in Region II produced a pink phenotype. Insertions in Region III resulted in either a light-yellow or a non-pigmented phenotype. Minicell studies showed that the 6.7 kb DNA fragment encoded at least five proteins (50 kDa, 42 kDa, 36 kDa, 35 kDa and 34 kDa). A 2.7 kb HindIII deletion in Region I caused the disappearance of these proteins, suggesting that this 2.7 kb fragment may play a regulatory role in pigment synthesis. Our results also showed that a 4.1 kb EcoRV fragment consisted of Region I and a part of Region II complemented a pink/yellow clone of Eho10 (pHL545), suggesting that the pigments of Eho13 and Eho10 were probably similar or identical.

Bacterial Proteins↗

Differences in FM100-Hue test performance related to iris colour may be due to pupil size as well as presumed amounts of macular pigmentation.

BACKGROUND: It has been reported that the differences in macular pigment between Asian and Caucasian eyes might give rise to different performance on the Farnsworth-Munsell 100 Hue test. However, the study did not appear to account adequately for the observation that differences were apparent only in older age groups. It was also acknowledged that other factors, like pupil size and crystalline lens colouration, could influence the result. METHOD: We investigated the performance of the Farnsworth-Munsell 100 Hue test, under standard conditions, by three groups with presumed different macular pigmentation and narrow age range. We also measured pupil size. RESULTS: The group with blue irides (presumed low macular pigmentation) performed significantly better on the colour vision task than the Asian group and the brown-eyed subjects as a whole (presumed higher macular pigmentation). In addition, the difference in pupil size between the blue-eyed group and brown-eyed groups bordered on significance. The difference in pupil size and consequential reduction in retinal illuminance account for a significant proportion of the difference in colour vision performance. In addition, a difference in both pupil size and Farnsworth-Munsell 100 Hue test performance was also observed between males and females that reinforced the importance of pupil size, and not just macular pigment, as a factor in colour vision performance difference between Asian and non-Asian eyes. CONCLUSION: It is concluded that pupil size, not just macular pigment, may have a significant influence on colour vision performance in a young, healthy population.

Adult↗

Incidence of corneal pigmented arc and factors associated with its appearance in orthokeratology.

PURPOSE: To determine the incidence of the corneal pigmented arc in orthokeratology (ortho-k) lens wearers over 12 months of lens wear and the factors associated with its appearance. METHOD: Thirty-five ortho-k subjects were recruited; refractive and corneal changes after lens wear (single-lens protocol) were monitored over 12 months. The incidence of the pigmented arc after 3, 6 and 12 months of lens wear was determined. RESULTS: The incidence of corneal pigmented arc was 17% (27%), 49% (49%) and 90% (93%) after 3, 6 and 12 months lens wear respectively in the left and right eyes. For subjects with arcs observed in the left eye within the first 6 months of lens wear, the mean +/- S.D. period of lens wear before initial detection of the arc was 14 +/- 7.4 weeks, and no correlation was found between this factor and the baseline spherical and cylindrical refractive errors (i.e. refractive sphere and cylinder, respectively), spherical equivalent refractive error (SERE), the target myopia reduction, the amount of refractive sphere (or SERE) reduction and changes in central and peripheral corneal curvatures after 6 months of lens wear. Baseline refractive sphere, baseline SERE, target, amount of myopia reduction, and change in central corneal curvature were significantly larger (p < 0.05) in those subjects with pigmented arcs after about 6 months of lens wear. The intensity of the observed pigmented arcs after about 6 months of lens wear was significantly related to the time when it was first observed (p = 0.003). Significant correlation was also found between the intensity of the arcs and the following parameters: baseline refractive sphere and SERE, target, change in central corneal curvature, and amount of myopia reduction (p < 0.006). After about 12 months of lens wear, the intensity of observed arcs was significantly related to the baseline refractive sphere, SERE and the target (p < 0.006). CONCLUSION: The incidence of ortho-k-associated pigmented arc increases from 17% after 3 months of lens wear to over 90% after 12 months of lens wear. The intensity of the arc is related to the time when the arc first appeared. Both the incidence and the intensity of the arc are related to the period of lens wear, baseline refractive sphere, SERE and the target.

Adolescent↗

Aggregation of pigment granules in single cultured Xenopus laevis melanophores by melatonin analogues.

1. Isolated melanophores were differentiated from aggregates of neural crest obtained from neurula stage Xenopus laevis embryos after 2 days in culture. 2. Condensation of pigment granules in these cells by melatonin (5-methoxy N-acetyltryptamine, aMT) and various novel analogues was monitored with an image analysis system to quantitate the area occupied by pigment in individual cells. 3. Melanophores exposed to vehicle (a maximum of 0.1% MeOH) showed little (less than 5%) change in pigment area. aMT produced a dramatic condensation of pigment granules (EC50 = the concentration producing a half maximal condensation, 9 pM). The response was rapid, reached a maximum (approximately 80% decrease in pigmented area) by 10 min, and was reversible after removal of aMT from the culture medium. 4. Aggregation to aMT was blocked by treating melanophores with pertussis toxin (1 microgram ml-1, 7 h) indicating a role for a guanosine 5' triphosphate (GTP)-binding protein in transducing the aMT receptor signal. 5. Structure-activity studies indicated that analogues of aMT lacking a side-chain N-acyl substituent (5-methoxytryptamine, MT) or a group at the 5-position of the indole ring (N-acetyltryptamine, aT) were unable to induce pigment aggregation (EC50 greater than 10 microM). 6. Lengthening the side-chain N-acyl group (N-propionyl, N-butanoyl) was tolerated to some degree but eventually (N-valeroyl and larger) activity diminished. Of the 5-position analogues tested 5-methoxy (aMT) was by far the most potent. 7. Halogen substitution in the 6-position of the indole ring led to some loss of activity as did a 6-OH substitution. The 6-OCH3 compound was inactive.8. These studies demonstrate the utility of this model in investigations of structure-activity relationships at the aMT receptor and suggest that it may be a valuable system for determining the transduction mechanisms coupled to the aMT receptor.

Animals↗

Use of the Q-switched 755-nm alexandrite laser to treat recalcitrant pigment after depigmentation therapy for vitiligo.

BACKGROUND: In patients with extensive vitiligo, depigmentation therapy is often preferred over attempts to restore skin color. Bleaching creams sometimes fail to permanently depigment the skin, leaving disfiguring pigmented patches. This recalcitrant pigmentation may be treated with Q-switched laser technology. OBJECTIVE: The objective was use the Q-switched alexandrite laser to treat recalcitrant pigmentation after unsuccessful attempts at total-body depigmentation for vitiligo. METHODS: We have attempted to destroy the remaining melanin pigmentation in a 68-year-old woman with vitiligo by using selective photothermolysis from a Q-switched alexandrite laser. RESULTS: Excellent results were achieved after 10 treatment sessions, with no recurrence of pigment after 1 year of follow-up. CONCLUSION: The Q-switched alexandrite laser was used successfully and safely in a vitiligo patient with recalcitrant pigmentation after unsuccessful depigmentation therapy and may prove useful in other such cases.

Aged↗

Effect of skin pigmentation on pulse oximetry accuracy in the emergency department.

OBJECTIVE: To determine whether pulse oximeter (PO) accuracy and signal quality are affected by level of skin pigmentation. METHODS: Observational study in a community hospital ED. Consecutive adult patients undergoing arterial blood gas determination were enrolled into the study. Skin pigmentation was determined by comparison with standardized color swatches under controlled lighting; assigned values were used to stratify patients into 3 groups (light, intermediate, and dark) using predetermined criteria. Simultaneous with arterial blood sampling, staff recorded PO reading of O2 saturation using the Nellcor D-25 oximeter. PO values were compared with criterion standard values measured using a 4-wavelength spectrophotometer or co-oximeter. PO signal quality also was recorded. Bias (the mean difference between PO and co-oximeter-measured values of hemoglobin saturation) and precision (the standard deviation of the bias) were calculated. Groups were compared using one-way ANOVA, Bartlett's test for variances, and chi2 test. RESULTS: O2 saturation data were obtained for 284 patients. Bias values did not differ between the 3 skin pigment groups (p = 0.79). Precision was of borderline significance (p = 0.05), but there was no dose-response relation between skin pigmentation and precision. Study personnel reported suboptimal PO function most often among patients in the dark group (p = 0.003), but this finding was of no clinical significance. PO signal failure was rare (<1% of all patients). CONCLUSIONS: Although several prior studies suggest the contrary, this study found that skin pigmentation does not affect the bias or precision of pulse oximetry. Furthermore, skin pigmentation has no clinically significant effect on PO signal quality.

Adult↗

Anaerobic breakdown of uroporphyrins I and III to bile pigments by extracts of Clostridium tetanomorphum.

Two blue bile pigments were formed under anaerobic conditions from the tetrapyrrole precursor delta-aminolevulinate by cells and cell extracts of Clostridium tetanomorphum. These compounds were also formed by cell extracts from the octacarboxylic tetrapyrrole, uroporphyrin III. Bactobilin, the first bacterial bile pigment to be discovered, is related to uroporphyrin I. The present results hence increase the number of bile pigments related to bactobilin. Bactobilin and its isomers differ markedly from the eukaryotic bile pigments which are all related to the dicarboxylic compound, protoporphyrin IX. The enzyme participating in the formation of the bacterial bile pigments was obligatorily anaerobic, in decided contrast to the only other known bile pigment-forming enzyme, the eukaryotic oxygen-requiring heme oxygenase.

Aminolevulinic Acid↗

Occupational pigmented contact dermatitis from Naphthol AS.

Hyper-pigmentation as a manifestation of contact sensitivity to optical brighteners has previously been reported. In 1973 we observed an epidemic of pigmented contact dermatitis occurring in a textile mill in connection with the manufacture of drill fabrics impregnated with a recently introduced azo dye coupling component Naphthol AS. Of the 53 workers included in the study, 12 presented a spotted pattern of hyperpigmentation mainly affecting the exposed areas, but without pruritus, 18 were pigmented to a lesser degree, and in 21 it was not possible to discern any well-defined discoloration; some of the patients in the last group, however, complained of slight pruritus. The last two patients who were fair-skinned showed a reddish-brown pigmentation and a well pronounced pruritus. Hyper-pigmentation was most pronounced in individuals with dark complexions, whereas fair-skinned patient also showed "classical" eczematous symptoms including pruritus. A field study demonstrated direct exposure to the chemical. All 53 patients were patch-tested with Naphthol AS 5% in water, and 24 were found to be positive. Histological features were melanocyte proliferation, incontinence of pigment, and various degrees of damage to the basal layer.

Anilides↗

An ex vivo study of congenital pigmented nevi in epidermal reconstructs.

In order to study morphologic and functional characteristics of pigment cells in congenital pigmented nevi, autologous or heterologous reconstructs have been made using normal keratinocytes and nevus cells from the dermal-epidermal junction or from the dermis. All these cells, keratinocytes and nevus cells, were used as cell suspensions immediately after dissociation from the tissues or after subsequent brief cultivation in a serum-free medium. Reconstructed epidermis were cultured for 15 days at the air-liquid interface with or without ultraviolet (UV) B exposure. The reconstructs were examined macroscopically (formation of hyperpigmented macules), histologically (pigment cell nesting) and ultrastructurally (pigment structure and transfer). Typical nesting of nevus cells was observed in the dermal-epidermal junction or in the superficial dermis associated with macroscopically detectable small pigmented macules. UVB exposure induced an upward migration of nevus cells in the suprabasal layers of the epidermis. This tissue model can be considered as an excellent system for the ex vivo reproduction of pigmented nevi and as an assay of the sensitivity of nevus cells towards UVB irradiation.

Cells, Cultured↗

Molecular mechanisms of pigment transport in melanophores.

We present an overview of the research on intracellular transport in pigment cells, with emphasis on the most recent discoveries. Pigment cells of lower vertebrates have been traditionally used as a model for studies of intracellular transport mechanisms, because these cells transport pigment organelles to the center or to the periphery of the cell in a highly co-ordinated fashion. It is now well established that both aggregation and dispersion of pigment in melanophores require two elements of the cytoskeleton: microtubules and actin filaments. Melanosomes are moved along these cytoskeletal tracks by motor proteins. Recent studies have identified the motors responsible for pigment dispersion and aggregation in melanophores. We propose a model for the possible roles of the two cytoskeletal transport systems and how they might interact. We also discuss the putative mechanisms of regulation of pigment transport, especially phosphorylation. Last, we suggest areas of research that will receive attention in the future in order to elucidate the mechanisms of organelle transport.

Actins↗

Granulocyte-macrophage colony-stimulating factor (GM-CSF) controls the proliferation and differentiation of mouse epidermal melanocytes from pigmented spots induced by ultraviolet radiation B.

Repeated exposure of ultraviolet radiation B (UVB) on the dorsal skin of hairless mice induces the development of pigmented spots long after its cessation. The proliferation and differentiation of epidermal melanocytes in UVB-induced pigmented spots are greatly increased, and those effects are regulated by keratinocytes rather than by melanocytes. However, it remains to be resolved what factor(s) derived from keratinocytes are involved in regulating the proliferation and differentiation of epidermal melanocytes. In this study, primary melanoblasts (c. 80%) and melanocytes (c. 20%) derived from epidermal cell suspensions of mouse skin were cultured in a basic fibroblast growth factor-free medium supplemented with granulocyte-macrophage colony-stimulating factor (GM-CSF). GM-CSF induced the proliferation and differentiation of melanocytes in those keratinocyte-depleted cultures. Moreover, an antibody to GM-CSF inhibited the proliferation of melanoblasts and melanocytes from epidermal cell suspensions derived from the pigmented spots of UV-irradiated mice, but not from control mice. Further, the GM-CSF antibody inhibited the proliferation and differentiation of melanocytes co-cultured with keratinocytes derived from UV-irradiated mice, but not from control mice. The quantity of GM-CSF secreted from keratinocytes derived from the pigmented spots of UV-irradiated mice was much greater than that secreted from keratinocytes derived from control mice. Moreover, immunohistochemistry revealed the expression of GM-CSF in keratinocytes derived from the pigmented spots of skin in UV-irradiated mice, but not from normal skin in control mice. These results suggest that GM-CSF is one of the keratinocyte-derived factors involved in regulating the proliferation and differentiation of mouse epidermal melanocytes from UVB-induced pigmented spots.

Animals↗

Regulation of pigmentation in zebrafish melanophores.

In comparison with the molecular genetics of melanogenesis in mammals, the regulation of pigmentation in poikilothermic vertebrates is poorly understood. Mammals undergo morphological colour change under hormonal control, but strikingly, many lower vertebrates display a rapid physiological colour change in response to the same hormones. The recent provision of extensive genome sequencing data from teleost zebrafish, Danio rerio, provides the opportunity to define the genes and proteins mediating this physiological pigment response and characterise their function biologically. Here, we illustrate the background adaptation process in adults and larvae and describe a novel assay to visualize and directly quantify the rate of zebrafish melanophore pigment translocation in unprecedented detail. We demonstrate the resolution of this assay system; quantifying the zebrafish melanophore response to melanin-concentrating and melanocyte-stimulating hormones. Furthermore, we investigate the intracellular signalling downstream of hormone stimulation and the biomechanical processes involved in zebrafish pigment translocation, confirming the importance of cyclic adenosine monophosphate (cAMP) as a mediator of pigment translocation and finding intact microtubules are essential for both melanin dispersion and aggregation in zebrafish, but that microfilament disruption affects aggregation only. In conclusion, we propose these data establish the zebrafish as an experimental model for studying both physiological colour change and the molecular basis of pigment translocation.

Actin Cytoskeleton↗

Skin pigmentation in Caucasian babies is high and evenly distributed throughout the body.

To investigate if Caucasian babies have particular sun sensitive skin and if skin pigmentation before any sun exposure is uniform throughout the body, we measured skin pigmentation objectively by skin reflectance spectroscopy in 10 anatomical sites in 20 healthy Caucasian babies (mean age 5 months, range 1 to 10 months) that had not been sun exposed previously. We found that skin pigmentation at all the measured sites was significantly higher than the constitutive pigmentation in Caucasian adults (P < 0.01). Furthermore, the level of skin pigmentation in all the 10 measured sites in the babies was statistically not different (P < 0.31) and there was no gender differences in pigmentation for any site.

Adult↗

Iodopsin, a red-sensitive cone visual pigment in the chicken retina.

The vertebrate retina contains two kinds of visual cells: rods, responsible for twilight (scotopic) vision (black and white discrimination); and cones, responsible for daylight (photopic) vision (color discrimination). Here we attempt to explain some of their functional differences and similarities in terms of their visual pigments. In the chicken retina there are four types of single cones and a double cone; each of the single cones has its own characteristic oil droplet (red, orange, blue, or colorless) and the double cone is composed of a set of principal and accessory members, the former of which has a green-colored oil droplet. Iodopsin, the chicken red-sensitive cone visual pigment, is located at outer segments of both the red single cones and the double cones, while the other single cones and the rod contain their own visual pigments with different absorption spectra. The diversity in absorption spectra among these visual pigments is caused by the difference in interaction between chromophore (11-cis retinal) and protein moiety (opsin). However, the chromophore-binding pocket in iodopsin is similar to that in rhodopsin. The difference in absorption maxima between both pigments could be explained by the difference in distances between the protonated Schiff-bases at the chromophore-binding site and their counter ions in iodopsin and rhodopsin. Furthermore, iodopsin has a unique chloride-binding site whose chloride ion serves for the red-shift of the absorption maximum of iodopsin. Visual pigment bleaches upon absorption of light through several intermediates and finally dissociates into all-trans retinal and opsin. That the sensitivity of cones is lower than rods cannot be explained by the relative photosensitivity of iodopsin to rhodopsin, but may be understood to some extent by the short lifetime of an enzymatically active intermediate (corresponding to metarhodopsin II) produced in the photobleaching process of iodopsin. The rapid formation and decay of the meta II-intermediate of iodopsin compared with metarhodopsin II are not contradictory to the rapid generation and recovery of cone receptor potential compared with rod receptor potential. The rapid recovery of the cone receptor potential may be due to a more effective shutoff mechanism of the visual excitation, including the phosphorylation of iodopsin. The rapid dark adaptation of cones compared with rods has been explained by the rapid regeneration of iodopsin from 11-cis retinal and opsin. One of the reasons for the rapid regeneration and susceptibility to chemicals of iodopsin compared with rhodopsin may be a unique structure near the chromophore-binding site of iodopsin.

Amino Acid Sequence↗

Role of hydroxyl-bearing amino acids in differentially tuning the absorption spectra of the human red and green cone pigments.

The human red and green cone pigments differ at either 15 or 16 amino acids, depending upon which polymorphic variants are compared. Seven of these amino acid differences involve the introduction or removal of a hydroxyl group. One of these differences, a substitution of alanine for serine at position 180, was found previously to produce a 5 nm blue shift. To determine the role of the remaining six hydroxyl group differences in tuning the absorption spectra of the human red and green pigments, we have studied six site-directed mutants in which single amino acids from the green pigment have been substituted for the corresponding residues in the red pigment. Blue shifts of 7 and 14 nm were observed upon substitution of phenylalanine for tyrosine at position 277 and alanine for threonine at position 285, respectively. Single substitutions at positions 65, 230, 233, and 309 produced spectral shifts of 1 nm or less. These data are in good agreement with a model based upon sequence comparisons among primate pigments and with the properties of site-directed mutants of bovine rhodopsin. Nonadditive effects observed in comparing the absorption spectra of red-green hybrid pigments remain to be explained.

Cell Line↗

Ocular distribution of topically applied adrenaline in albino and pigmented rabbits.

Systemic absorption, ocular distribution and ocular metabolism in albino and pigmented rabbit of topically applied 1% [3H]l-adrenaline eye drops was studied by liquid scintillation counting and thin layer chromatography. The effect of adrenaline on the pupillary diameter was also registered. The peak plasma level of adrenaline was reached at 150 min. The corneal adrenaline metabolism was substantial, but did not vary with the rabbit strain. At 30 min after instillation, the total adrenaline concentration of pigmented iris and ciliary body exceeded that of corresponding albino tissues. At 180 min the drug concentration of the iris and ciliary body did not differ between the rabbit strains. The time course of adrenaline induced mydriasis was equal in both rabbit strains. Since neither total adrenaline concentration at 180 min nor the time course of adrenaline induced mydriasis differed between the rabbit strains, the higher initial adrenaline uptake by pigmented tissues is not explained by pigment binding. It is proposed that the increased initial binding of adrenaline by pigmented tissues is caused by a difference between pigmented and albino tissues in the number of adrenergic neurones; this is also supported by the literature.

Albinism↗

Acute posterior multifocal placoid pigment epitheliopathy.

Five patients with acute posterior multifocal placoid pigment epitheliopathy (APMPPE) have been subject to a follow-up study. All the patients had bilateral lesions. The follow-up period ranged from 1 month to 8 years (mean 4 years). All had resumed their previous work, although two patients had severe visual loss in one eye (6/36). No recurrences were observed. Automatic perimetry of the central visual field revealed moderate central scotomata between 5 and 10 db, corresponding to derangement of the pigment epithelium. This may indicate that the pigment derangement may be caused by alterations in the pigment granules in the pigment epithelial cells, and that the pigment epithelial cells have recovered. There were absolute scotomata in two eyes corresponding to chorioretinal atrophy.

Acute Disease↗

Incremental responses to light recorded from pigment epithelial cells and horizontal cells of the cat retina.

1. Rod-dependent incremental responses were recorded intracellularly in both pigment epithelial cells and horizontal cells of the cat retina. They were elicited by test flashes which were superimposed on background flashes after a delay.2. In pigment epithelial cells smaller test responses were produced as background intensity was raised. The incremental sensitivity function was linear for about 1.4 log units, with a slope of 0.86, and the approach of saturation occurred at about 2.5 log td scotopic.3. The amplitude of pigment epithelial test responses could be estimated from the dark-adapted amplitude-log intensity function obtained with single flashes. Test flashes produced the voltage increment predicted by the slope of this function just above the point on the curve equal to the background intensity. The pigment epithelial response to a test flash, therefore, is the response expected if the background were presented alone and made more intense by the amount of the test flash.4. Rod-dependent incremental sensitivity functions of horizontal cells closely resembled the ones obtained from pigment epithelial cells.5. It was concluded that the adaptive effects observed in pigment epithelial cells originated in individual rods. These effects arose from the compressive nature of the dark-adapted amplitude-intensity function. In horizontal cell responses these effects may be modified by the failure of the background response to maintain its initial voltage.

Animals↗