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PubMed2000

Depigmentation of the normally pigmented patches in universal vitiligo patients by cryotherapy.

BACKGROUND: Complete depigmentation may occur, albeit rarely, in patients with universal vitiligo. They usually have one or more pigmented patches that remain normal, most frequently over the malar area of the face and dorsal aspects of forearms. Total repigmentation may not occur in these patients, even after receiving 150 session of PUVA therapy alone or in conjunction with other medical or surgical procedures, and there is possibility of recurrence after cessation of therapy. These patients are usually more interested in depigmentation rather than trying for repigmentation. OBJECTIVE: Because of the relatively high sensitivity of melanocytes to cryotherapy and the possibility of isomorphic phenomenon in vitiligo patients, removal of the remaining normally pigmented patches was attempted in patients with universal vitiligo using cryotherapy. METHOD: Five volunteer patients with universal vitiligo were chosen, all of whom wished to have their pigmented patches removed. In all five patients a small area was tested first in order to assure the patients of what the procedure involved and to show its likely result. All pigmented areas were then treated to 1-3 sessions of cryotherapy using a closed contact CO2 cryogun, with 4-6-week intervals. RESULTS: Complete and permanent depigmentation was achieved in all five patients with excellent cosmetic results and no complications or scarring. CONCLUSION: Cryotherapy is a cost effective, non-complicating, easily available procedure which can be used for depigmentation of normally pigmented patches in patients with universal vitiligo.

Adult↗
PubMed2000

Effect of constitutional pigmentation on ultraviolet B-induced DNA damage in fair-skinned people.

Ultraviolet light has been implicated as a dominant factor in skin cancer development. Skin pigmentation is traditionally regarded as an important protection against skin cancer. Yet, little is known about how skin pigmentation is modulating induction of DNA damage, which is the primary event in carcinogenesis. We applied a recently developed 32P-postlabeling technique to measure the effect of constitutional pigmentation on the formation of major ultraviolet-induced DNA damage in human skin in vivo. The induction of photoproducts showed a statistically significant negative correlation with erythemal response and skin pigmentation. Our results demonstrated that the constitutional pigmentation is efficiently guarding DNA against the formation of photoproducts. The difference in melanin content is likely to be one of the reasons for the observed interindividual variation in levels of DNA damage after the uniform exposure to ultraviolet B.

Adult↗
PubMedDate not supplied

Regulation of cutaneous pigmentation by titration of human melanocytes in cultured skin substitutes grafted to athymic mice.

Pigmentation of healed cultured skin substitutes in burn patients is frequently irregular and unpredictable which compromises solar protection and the patient's self-image. To address these morbidities, human fibroblasts were inoculated on a collagen-glycosaminoglycan substrate followed 1 day later by the addition of keratinocytes at 1.1 x 10(6)/cm2 combined with either 0, 1.1 x 10(2), 1.1 x 10(3), or 1.1 x 10(4) melanocytes/cm2. The skin substitutes were incubated in vitro for 3 weeks and grafted to athymic mice. In vitro, the number of L-Dopa-positive melanocytes in the skin substitutes increased proportionately to the number of melanocytes inoculated. The melanocytes localized to the basal epidermis when labeled for MEL-5. The skin substitutes with 1.1 x 10(4) melanocytes/cm2 were significantly darker than other groups in vitro by chromameter evaluation. By 12 weeks after grafting, the cultured skin ranged from no pigment in the control group, to 75% pigmented area in the 1.1 x 10(3) melanocytes/cm2 group, to complete pigmentation in the 1.1 x 10(4) melanocytes/cm2 group. In vivo, the mean chromameter values were significantly darker for the grafts with 1.1 x 10(3) and 1.1 x 10(4) melanocytes/cm2. These results suggest that complete restoration of cutaneous pigmentation can be accomplished by addition of between 0.1 and 1.0 x 10(4) melanocytes/cm2 to skin substitutes.

Animals↗
PubMed2001

[Autologous subretinal transplantation of cultivated porcine iris pigment epithelial cells (IPE)].

BACKGROUND: Subretina transplantation of epithelium may be a therapeutic option for surgical treatment of age-related macular degeration (AMD). Various experimental data have demonstrated that homologous transplantation of retinal pigment epithelium (RPE) can prevent photoreceptor deterioration. However, most investigators experienced immunogenic graft rejection when using homologous pigmented cells for grafting. Autologous cells were soon considered as an alternative for subretinal grafting. Particularly iris pigment epithelium (IPE) appeared suitable to replace homologous RPE for it embryogenetic similarity and its simple availability. Recent studies have shown, that IPE is capable of taking over functions of RPE in maintaining retinal metabolism. the purpose of this study was to evaluate if autologous IPE cells would survive when being transplanted subretinally. In addition, immunogenic reponses to the presence of "foreign" iris pigment cells needed to be excluded. MATERIALS AND METHODS: Iris tissue was obtained by peripheral iridectomy in the anesthetized pig. Sheets of pigment iris epithelium were separated from the specimens and transferred into tissue culture. After the cells had been grown to confluency, cell suspensions were injected into the subretinal space of the donor animal's fellow eye. After 4 weeks, the grafted eye was enucleated and examined histologically.. RESULTS: The histological exam revealed that the graft cells had survived in the subretinal space. No evidence of immunogenic rejection was observed. CONCLUSION: Autologous IPE-cells can survive in the host's sub-retinal space without creating inflammatory reactions. Transplanted IPE appears to interact with photoreceptor outer segments.

Animals↗
PubMed1987

Prevalence and importance of pigmented ocular fundus lesions in Gardner's syndrome.

We examined 134 members of 16 families with Gardner's syndrome for pigmented ocular fundus lesions. Of 41 patients with documented Gardner's syndrome, 37 (90.2 percent) had such lesions. The lesions were bilateral in 32 of the patients (78.1 percent) and in 2 of 42 controls (4.8 percent). Twenty (46.5 percent) of 43 first-degree relatives at 50 percent risk for Gardner's syndrome had bilateral pigmented fundus lesions, indicating that they had probably inherited the abnormal gene. The presence of bilateral lesions, multiple lesions (more than four), or both appeared to be a specific (specificity, 0.952) and sensitive (sensitivity, 0.780) clinical marker for Gardner's syndrome. The lesions are probably congenital; they were observed in a three-month-old baby at risk. The multiplicity of the pigmented fundus lesions and their association with diffuse disturbances of the retinal pigment epithelium in the same eye suggest a widespread expression of the abnormal gene in the retinal pigment epithelial cells.

Female↗
PubMed2001

Lipofuscin pigmentation in pleomorphic adenoma of the palate.

This case of pleomorphic adenoma containing pigmented cells will contribute to the knowledge about pigmentation in salivary gland tumors because, to date, only a few such descriptions have been published. The pigment was examined by using histochemical and immunohistochemical methods, as well as by transmission electron microscopy. Pigment granules stained yellowish-brown with hematoxylin and eosin. The granules stained positive with Schmorl's, Ziehl-Neelsen, periodic acid-Schiff, Fontana-Masson, or Nile blue stains. Fluorescent microscopy revealed strong granule autofluorescence. Transmission electron microscopy disclosed cytoplasmic granules with homogeneous electron-dense appearance, ranging from 0.5 to 1.0 microm in dimension and arranged in diffusely distributed and aggregated patterns. The pigment was found to be lipofuscin, and the cells containing the granules were discovered to be plasmacytoid myoepithelial cells.

Adenoma, Pleomorphic↗
PubMed1994

The use of pigmentation and shading information in recognising the sex and identities of faces.

An investigation of what can be learned about representational processes in face recognition from the independent and combined effects of inverting and negating facial images is reported. In experiment 1, independent effects of inversion and negation were observed in a task of identifying famous faces. In experiments 2 through 4 the question of whether effects of negation were still obtained when effects due to the reversal of pigmentation in negative images were eliminated was examined. By the use of images of the 3-D surfaces of faces measured by laser, and displays as smooth surfaces devoid of pigmentation, only effects of inversion were obtained reliably, suggesting that the effects observed in experiment 1 arose largely through the inversion of pigmentation values in normal images of faces. The results of experiment 5 suggested that the difference was not due to the different task demands of experiments 2-4 compared with those of experiment 1. When normally pigmented face images were used in a task making similar demands to that of experiment 4, independent effects of inversion and negation were again observed. When a task of sex classification was used in experiments 6 and 7, clear effects of negation as well as inversion were observed on latencies, though not accuracies, of responding. The results are interpreted in terms of the information content of pigmentation relative to shape from shading in different face-classification tasks. The results also reinforce other recent evidence demonstrating the importance of image intensity as well as spatial layout of face 'features'.

Adult↗
PubMed1996

Selective transcription factor induction in retinal pigment epithelial cells during photoreceptor phagocytosis.

Expression of early response genes during rod outer segment phagocytosis by normal Long Evans and Royal College of Surgeons-rdy+p+ rats and by dystrophic Royal College of Surgeons-p+ rat retinal pigment epithelial cells was studied in primary cell culture. Northern analysis revealed that the abundance of zif-268 (egr-1), c-fos, and tis-1 (NGF1-B) mRNA was rapidly and transiently increased in normal retinal pigment epithelial cells during rod outer segment phagocytosis but not during phagocytosis of latex particles. No increase in gene expression was found in Royal College of Surgeons-p+ dystrophic retinal pigment epithelial cells challenged with rod outer segments. As shown by electrophoretic mobility shift assay, a prominent short term increase in the intensity of the gel-shifted band was detected using nuclear protein extracts derived from rod outer segment-challenged, control retinal pigment epithelial cells and zif-268, AP-1, AP-2, or tis-1 consensus oligonucleotides. No such increase was detected when using nuclear factor kappaB consensus oligonucleotide or when the early response gene prostaglandin H synthase-2 mRNA was measured over the time course studied. The results suggest that in retinal pigment epithelial cells, rod outer segment-specific phagocytosis is accompanied by the selective expression of early response genes coding for transcription factors. The specific pattern of the induction of these transcription factors is predicted to modulate the expression of gene cascades.

Animals↗
PubMed1998

Mechanisms of spectral tuning in blue cone visual pigments. Visible and raman spectroscopy of blue-shifted rhodopsin mutants.

Spectral tuning by visual pigments involves the modulation of the physical properties of the chromophore (11-cis-retinal) by amino acid side chains that compose the chromophore-binding pocket. We identified 12 amino acid residues in the human blue cone pigment that might induce the required green-to-blue opsin shift. The simultaneous substitution of nine of these sites in rhodopsin (M86L, G90S, A117G, E122L, A124T, W265Y, A292S, A295S, and A299C) shifted the absorption maximum from 500 to 438 nm, accounting for 2,830 cm-1, or 80%, of the opsin shift between rhodopsin and the blue cone pigment. Raman spectroscopy of mutant pigments shows that the dielectric character and architecture of the chromophore-binding pocket are specifically altered. An increase in the number of dipolar side chains near the protonated Schiff base of retinal increases the ground-excited state energy gap via long range dipole-dipole Coulomb interaction. In addition, the W265Y substitution causes a decrease in solvent polarizability near the chromophore ring structure. Finally, two substitutions on transmembrane helix 3 (A117G and E122L) act in combination with the other substitutions to alter the binding-pocket structure, resulting in stronger interaction of the protonated Schiff base group with the surrounding dipolar groups and the counterion. Taken together, these results identify the amino acid side chains and the underlying physical mechanisms responsible for a majority of the opsin shift in blue visual pigments.

Amino Acid Sequence↗
PubMed2002

Pigment binding of photosystem I light-harvesting proteins.

Light-harvesting complexes (LHC) of higher plants are composed of at least 10 different proteins. Despite their pronounced amino acid sequence homology, the LHC of photosystem II show differences in pigment binding that are interpreted in terms of partly different functions. By contrast, there is only scarce knowledge about the pigment composition of LHC of photosystem I, and consequently no concept of potentially different functions of the various LHCI exists. For better insight into this issue, we isolated native LHCI-730 and LHCI-680. Pigment analyses revealed that LHCI-730 binds more chlorophyll and violaxanthin than LHCI-680. For the first time all LHCI complexes are now available in their recombinant form; their analysis allowed further dissection of pigment binding by individual LHCI proteins and analysis of pigment requirements for LHCI formation. By these different approaches a correlation between the requirement of a single chlorophyll species for LHC formation and the chlorophyll a/b ratio of LHCs could be detected, and indications regarding occupation of carotenoid-binding sites were obtained. Additionally the reconstitution approach allowed assignment of spectral features observed in native LHCI-680 to its components Lhca2 and Lhca3. It is suggested that excitation energy migrates from chlorophyll(s) fluorescing at 680 (Lhca3) via those fluorescing at 686/702 nm (Lhca2) or 720 nm (Lhca3) to the photosystem I core chlorophylls.

Binding Sites↗
PubMed2004

Pigment binding, fluorescence properties, and oligomerization behavior of Lhca5, a novel light-harvesting protein.

A new potential light-harvesting protein, named Lhca5, was recently detected in higher plants. Because of the low amount of Lhca5 in thylakoid membranes, the isolation of a native Lhca5 pigment-protein complex has not been achieved to date. Therefore, we used in vitro reconstitution to analyze whether Lhca5 binds pigments and is actually an additional light-harvesting protein. By this approach we could demonstrate that Lhca5 binds pigments in a unique stoichiometry. Analyses of pigment requirements for light-harvesting complex formation by Lhca5 revealed that chlorophyll b is the only indispensable pigment. Fluorescence measurements showed that ligated chlorophylls and carotenoids are arranged in a way that allows directed energy transfer within the light-harvesting complex. Reconstitutions of Lhca5 together with other Lhca proteins resulted in the formation of heterodimers with Lhca1. This result demonstrates that Lhca5 is indeed a protein belonging to the light-harvesting antenna of photosystem I. The properties of Lhca5 are compared with those of previously characterized Lhca proteins, and the consequences of an additional Lhca protein for the composition of the light-harvesting antenna of photosystem I are discussed in view of the recently published photosystem I structure of the pea.

Arabidopsis↗
PubMed1998

Dedifferentiation of the retinal pigment epithelium compared to the proliferative membranes of proliferative vitreoretinopathy.

PURPOSE: To examine the gene expression for melanogenesis of retinal pigment epithelial cells during dedifferentiation and to compare the condition to that of eyes obtaining anatomical success after surgery for proliferative vitreoretinopathy. METHODS: Gene expression for melanogenesis was determined by reverse transcriptase-polymerase chain reaction of tyrosinase and tyrosinase-related protein-1 genes in normal and cultured retinal pigment epithelial cells and in proliferative membranes in patients with proliferative vitreoretinopathy. RESULTS: Gene expression for melanogenesis was classified into three types during dedifferentiation of retinal pigment epithelial cells: (1) tyrosinase-related protein-1 gene expression, (2) tyrosinase and tyrosinase-related protein-1 gene expression and (3) no expression of these genes. The expression of these genes were maintained better in mediums with basic fibroblast growth factor than in medium without basic fibroblast growth factor. Of the anatomically unsuccessful patients with proliferative vitreoretinopathy treated by surgery, 76.9% showed both tyrosinase and tyrosinase-related protein-1 gene expression; only 20% of the anatomically successful patients showed the gene expression. CONCLUSIONS: We reported three different conditions of retinal pigment epithelial cells based on gene expression for melanogenesis during dedifferentiation. The different condition of the retinal pigment epithelial cells may have some relationship to the anatomical results for proliferative vitreoretinopathy surgery.

Adolescent↗
PubMed2003

Participation of pigment epithelium in ocular immune privilege. 3. Epithelia cultured from iris, ciliary body, and retina suppress T-cell activation by partially non-overlapping mechanisms.

PURPOSE: The ocular microenvironment is immunosuppressive and anti-inflammatory. Since various ocular pigmented epithelia contribute to this microenvironment, we studied the relative capacities of pigment epithelial (PE) cells cultured from the iris, ciliary body, and retina of mouse eyes to suppress T-cell activation in vitro. METHODS: Pigment epithelium was cultured from iris, ciliary body, and retina for 14 days, then assayed for the capacity, directly or across transwell membranes, to suppress mixed lymphocyte reactions and anti-CD3 stimulation of T cells. Potential molecules responsible for suppression were examined by attempting to block suppression with appropriate reagents, and by using mice with pertinent mutant or disrupted genes. RESULTS: We found that PE cells from all three ocular tissue sources profoundly suppressed T-cell activation in vitro. While iris PE suppressed poorly when separated from T cells by a transwell membrane (implying that cell contact is necessary), retina PE suppressed fully even in the presence of such a membrane (implying that soluble factors were responsible). Ciliary body PE used both soluble factors as well as cell contact to achieve suppression. Suppression could not be ascribed to TGFbeta, IFNgamma, TNFalpha, CD48, or ICAM-1, or to interactions between CD40 and CD154, or CD95 and CD95 ligand. Galectin-1, a galactoside-binding protein, was found to be expressed on all cultured PE cells, but only retinal pigment epithelium (RPE) from galectin-1 KO mice showed reduced capacity to inhibit T-cell activation. CONCLUSIONS: Cultured pigment epithelia from iris, ciliary body, and retina comparably suppress T-cell activation in vitro, but by partially different mechanisms. Although RPE cells suppress in part through expression of galectin-1, the molecular mediators of suppression by iris and ciliary body PE remain to be identified.

Animals↗
PubMed1975

Purification and some properties of malarial pigment.

Malarial pigment from erythrocytes infected with Plasmodium berghei was purified by treatment with sodium dodecyl sulphate solution, followed by incubation with Pancreatin. The purified pigment retained the apparently crystalline form of pigment within the parasite, rotated polarised light and had the same solubility characterisation as crude malarial pigment. It contained about 1% iron, all of which could be accounted for in terms of haemin. The iron of the pigment molecule is oxidised by the parasite to the ferric state.

Animals↗
PubMed2000

Brown pigment stones in the common bile duct: reduced bilirubinate diconjugate in bile.

BACKGROUND: Bilirubin is the main component of most common bile duct stones. Normally, almost all bilirubin in bile is conjugated to glucuronic acid or some other sugar moiety. These conjugates are unstable and liable to deconjugation. Unconjugated bilirubin is insoluble and may precipitate as the calcium salt found in brown pigment stones. The pattern of bilirubin conjugates in common duct bile of patients with choledocholithiasis has been unknown. METHODS: In a clinical series of 55 patients with choledocholithiasis common-duct bile was aspirated, and the bilirubin conjugates analyzed with high-performance liquid chromatography. One stone from each patient was analyzed for cholesterol and bilirubin content to determine stone type. RESULTS: Sixteen patients had cholesterol stones, 38 patients had brown pigment stones, and 1 patient had a black stone. Patients with pigment stones had a lower percentage of bilirubin diglucuronide (median, 60.3%; interquartile range, 49.7%-67.3%) than patients with cholesterol stones (64.0%; 60.2%-73.3%) (Mann-Whitney, P=0.015). No significant difference was found for the other bilirubin conjugates, total bilirubin, or biliary pH when pigment and cholesterol stone patients were compared. The time of bile sampling in relation to papillotomy and treatment of cholestasis was not associated with the low percentage of bilirubin diglucuronide. The observation of reduced values for bilirubin diglucuronide could not be ascribed to duodenal diverticula or Billroth-II gastric resection. CONCLUSION: The percentage of the main bilirubinate conjugate, bilirubin diglucuronide, is decreased in the common duct bile of patients with pigmented compared with cholesterol stones.

Aged↗
PubMed2003

Pulmonary toxicity studies in rats with triethoxyoctylsilane (OTES)-coated, pigment-grade titanium dioxide particles: bridging studies to predict inhalation hazard.

The aim of this study was to assess and compare the acute lung toxicities of intratracheally instilled hydrophobic relative to hydrophilic surface-coated titanium dioxide (TiO(2)) particles using a pulmonary bridging methodology. In addition, the results of these instillation studies were bridged with data previously generated from inhalation studies with hydrophilic, pigment-grade (base) TiO(2) particles, using the base, pigment-grade TiO(2) particles as the inhalation/instillation bridge material. To conduct toxicity comparisons, the surface coatings of base pigment-grade TiO(2) particles were made hydrophobic by application of triethoxyoctylsilane (OTES), a commercial product used in plastics applications. For the bioassay experimental design, rats were intratracheally instilled with 2 or 10 mg/kg of the following TiO(2) particle-types: (1) base (hydrophilic) TiO(2) particles; (2) TiO(2) with OTES surface coating; (3) base TiO(2) with Tween 80; or (4) OTES TiO(2) with Tween 80. Saline instilled rats served as controls. Following exposures, the lungs of sham- and TiO(2)-exposed rats were assessed both using bronchoalveolar lavage (BAL) biomarkers and by histopathology of lung tissue at 24 hours, 1 week, 1 month, and 3 months post exposure. The results demonstrated that only the base, high-dose (10 mg/kg) pigment-grade TiO(2) particles and those with particle-types containing Tween 80 produced a transient pulmonary inflammatory response, and this was reversible within 1 week postexposure. The authors conclude that the OTES hydrophobic coating on the pigment-grade TiO(2) particle does not cause significant pulmonary toxicity.

Animals↗
PubMed2006

Characteristics of iris and retinal pigment epithelial cells cultured on collagen type I membranes.

PURPOSE: Transplantation of pigment epithelial cells is a promising treatment modality to repair retinal damage in age-related macular degeneration. For this purpose, it is necessary to establish cell culture techniques that allow acquisition of proper functional and morphological characteristics by the cells to be transplanted. METHODS: Primary retinal pigment epithelial (RPE) and iris pigment epithelial (IPE) cells grown to confluence on collagen membranes were examined for morphology, adhesion, proliferation, apoptosis, as well as viability after ex vivo transplantation. RESULTS: Pigment epithelial cells adhere, proliferate, form monolayers, acquire differentiated properties, and remain viable during transplantation to the subretinal space. CONCLUSIONS: Pigment epithelial cells cultured on collagen membranes acquire differentiated characteristics and are amenable to be transplanted as cell monolayers.

Animals↗
PubMed1985

Isolation and characterization of a factor from calf serum that promotes the pigmentation of embryonic and transformed melanocytes.

A protein (Mr = 63,000) from calf serum that promotes the pigmentation of cultured chick neural crest and mouse melanoma cells has been partially isolated and characterized in this study. The stimulation of melanin synthesis in cultured cells was used to follow its activity during purification. The pigment-promoting factor was isolated by sequential column chromatography on dye-agarose matrices followed by hydroxyapatite and high pressure molecular sieve chromatography. The factor was found to stimulate melanin biosynthesis at 2-4 micrograms/ml and was specific for melanin-producing cells and their precursors. Antibodies raised in rabbits against the factor inhibited its pigment-promoting activity as well as that of whole calf serum. Enzyme-linked immunoadsorbent assays demonstrated that calf and bovine sera contain molecules that cross-react with the pigment-promoting factor. Horse, human, rat, and chicken sera, which lack the biological activity, also lacked immunological cross-reactivity. Extracts of certain tissues, particularly the submaxillary gland, were observed to be rich sources of pigment-promoting activity.

Animals↗