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Black pigment gallstones with cholesterol gallstones in the same gallbladder. 13 cases in a surgical series of 1226 patients with gallbladder stones.

We studied 1312 consecutive patients who underwent surgery for gallstones in the biliary tract at one university hospital in Siena, Italy, with a systematic classification of gallstones found within the gallbladder. Of these patients, 1226 were found to have gallbladder stones; 94 of these had black pigment gallstones. Of these, 13 patients were found to have black pigment gallstones and cholesterol gallstones within their gallbladder. They all had multiple black pigment gallstones, usually very small (all < 6 mm diameter), in association with larger cholesterol stones in the gallbladder lumen. The cholesterol gallstones were single in seven cases, double in two cases, and multiple in four cases. All 13 of these patients with black pigment stones in association with cholesterol stones had histologic evidence of either adenomyomatosis or Rokitansky-Aschoff sinuses in the gallbladder wall. In nine of the 13 patients, the black pigment stones were located both in the gallbladder lumen and in close association with the gallbladder wall (in areas of adenomyomatosis or in Rokitanski-Aschoff sinuses). In the other four patients, the stones were found in close association with the gallbladder wall alone and not freely mobile within the gallbladder lumen. It is concluded that cholesterol stones and black pigment stones may be found in the same gallbladder. This association is infrequent with an incidence of 13 of 1226 (1.06%) in our series. There appears to be some relationship between the formation of the black pigment stones and the presence of adenomyomatosis or Rokitanski-Aschoff sinuses.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenomyoma↗

Pigment epithelium-derived factor prevents advanced glycation end products-induced monocyte chemoattractant protein-1 production in microvascular endothelial cells by suppressing intracellular reactive oxygen species generation.

AIMS/HYPOTHESIS: Monocytes and macrophages accumulate in the lesion of the diabetic retina, which are most likely involved in the progression of diabetic retinopathy. The levels of monocyte chemoattractant protein-1 (MCP-1) in vitreous fluids were associated with the severity of proliferative diabetic retinopathy. Recently, pigment epithelium-derived factor has been shown to be involved in the pathogenesis of proliferative diabetic retinopathy. However, a role of pigment epithelium-derived factor in monocyte recruitments in diabetic retinopathy remains to be elucidated. In this study, we investigated effects of purified pigment epithelium-derived factor on AGE-induced reactive oxygen species generation, MCP-1 mRNA up-regulation and protein production in human cultured microvascular endothelial cells. METHODS: The intracellular formation of reactive oxygen species was measured using the fluorescent probe CM-H(2)DCFDA. MCP-1 gene expression was analysed in quantitative reverse transcription-polymerase chain reaction. Monocyte chemoattractant protein-1 production by microvascular endothelial cells was measured with an ELISA system. RESULTS: AGE increased intracellular reactive oxygen species generation in microvascular endothelial cells. Pigment epithelium-derived factor inhibited the AGE-induced reactive oxygen species generation in a dose-dependent manner. An anti-oxidant, N-acetylcysteine, or pigment epithelium-derived factor completely prevented the AGE-induced up-regulation of MCP-1 mRNA contents as well as protein production in microvascular endothelial cells. CONCLUSIONS/INTERPRETATIONS: Pigment epithelium-derived factor inhibits the AGE-induced reactive oxygen species generation and the subsequent increase in MCP-1 production in microvascular endothelial cells. Our study suggests that substitution of pigment epithelium-derived factor could prevent the progression of diabetic retinopathy by attenuating the deleterious effects of AGE.

Cells, Cultured↗

Low content of the natural ocular anti-angiogenic agent pigment epithelium-derived factor (PEDF) in aqueous humor predicts progression of diabetic retinopathy.

AIMS/HYPOTHESIS: Retinopathy is the most common microvascular complication of diabetes. Our aim was to address the predictive value of pro-angiogenic and anti-angiogenic markers for progression of retinopathy. METHODS: Aqueous humor was collected at cataract surgery from 32 diabetic patients who had no or very mild retinopathy (ETDRS stage </=20) and 33 normoglycaemic control subjects. Content of pro-angiogenic vascular endothelial growth factor and angiogenic inhibitor pigment epithelium-derived factor were determined. ANGIOGENIC activity was quantified by measuring its effect on the migration of capillary endothelial cells. The predictive value of the initial level of these markers for progression of retinopathy was studied by following the probands for a maximum of 75 months. RESULTS: In the aqueous fluid content of vascular endothelial growth factor was increased in diabetic patients (mean values 492 versus 292 pg/ml; p=0.0052), and pigment epithelium-derived factor values were decreased (mean values 1740 versus 3680 ng/ml; p=0.0058) compared to control subjects. Of the diabetic patients ten progressed during follow-up (ETDRS stage >47B). This subgroup showed lower pigment epithelium-derived factor content when compared to non-progressors and control subjects. Migratory activity in samples of patients from the control group and in diabetic patients without progression was generally inhibitory due to pigment epithelium-derived factor. Inhibition was blocked by neutralizing antibodies to pigment epithelium-derived factor. In diabetic patients initial angiogenic activity was higher in those who later developed retinopathy (vs. controls p=0.00005; vs. no progressors p=0.0003). Both pigment epithelium-derived factor and migratory response predicted progression. CONCLUSION/INTERPRETATION: Pigment epithelium-derived factor is an important negative regulator of angiogenic activity of aqueous humor. Its content in the aqueous humor of diabetic patients strongly predicts who among them will develop progression of retinopathy.

Aged↗

Tuning of photoreceptor function in three mantis shrimp species that inhabit a range of depths. I. Visual pigments.

Visual pigments in many animal species, including stomatopod crustaceans, are adapted to the photic environments inhabited by that species. However, some species occupy a diversity of environments as adults (such as a range of depths in the ocean), and a single set of visual pigments would not be equally adaptive for all habitats in which individuals live. We characterized the visual pigment complements of three species of stomatopod crustaceans, Haptosquilla trispinosa, Gonodactylellus affinis, and Gonodactylopsis spongicola, which are unusual for this group in that each lives at depths from the subtidal to several tens of meters. Using microspectrophotometry, we determined the visual pigments in all classes of main rhabdoms in individuals of each species from shallow or deep habitats. Each species expressed the typical diversity of visual pigments commonly found in stomatopods, but there was little or no evidence of differential expression of visual pigments in animals of any species collected from different depths. Vision in these species, therefore, is not tuned to spectral characteristics of the photic environment by varying the assemblages of visual pigments appearing in their retinas.

Animals↗

Effect of ultraviolet radiation on melanogenesis in four different types of cultured bovine ocular pigmented cells.

BACKGROUND: Ultraviolet radiation is thought to play a causative role in various ocular diseases such as macular degeneration, cataract, and possibly melanomas. Since most of the energy is absorbed by pigmented cells, the aim of this study was to examine and compare the reactions of different ocular melanocytic cells to ultraviolet light in vitro. MATERIALS AND METHODS: Bovine iris melanocytes, choroidal melanocytes, iris pigment epithelial cells, and retinal pigment epithelial cells were isolated and cultured. Semiconfluent cultures were exposed to ultraviolet radiation (280-380 nm). Cell number and melanin content were measured 10 days after radiation. Selected samples were examined by transmission electron microscopy. RESULTS: Following irradiation with ultraviolet light for 30 s, 60 s, and 120 s, the number of cells in culture decreased markedly. In contrast, total melanin content in the cultures of iris melanocytes, choroidal melanocytes, and iris pigment epithelial cells did not decrease despite the reduced number of cells. This finding suggested an increase in melanin per cell. However, the increase in average melanin content observed was not due to melanogenesis, because treatment with the melanogenesis inhibitor alpha-methyl-p-tyrosine did not reduce the melanin content of the cultures and electron-microscopic examination showed no morphological evidence of increased melanogenesis. CONCLUSION: In vitro, there was no convincing evidence of ultraviolet radiation-induced melanogenesis in ocular pigmented cells. Thus, it seems that ultraviolet radiation is a selection factor: more densely pigmented cells survive the treatment better than less pigmented cells.

Animals↗

Genetic analysis of steel and the PG-M/versican-encoding gene AxPG as candidates for the white (d) pigmentation mutant in the salamander Ambystoma mexicanum.

Vertebrate non-retinal pigment cells are derived from neural crest (NC) cells, and several mutations have been identified in the Mexican axolotl Ambystoma mexicanum (Ambystomatidae) that affect the development of these cell lineages. In "white" (d) mutant axolotls, premigratory NC cells differentiate as pigment cells, yet fail to disperse, survive, or both, and this leads to a nearly complete absence of pigment cells in the skin. Previous studies revealed that d affects pigment cell development non-autonomously, and have reported differences between white and wild-type axolotls in the structure and composition of the extracellular matrix through which NC and pigment cells migrate. Here we test the correspondence of d and two candidate genes: steel and AxPG. In amniotes, Steel encodes the cytokine Steel factor (mast cell growth factor; stem cell factor; kit ligand), which is expressed along the migratory pathways of melanocyte precursors and is required by these cells for their migration and survival; mammalian Steel mutants resemble white mutant axolotls in having a deficit or complete absence of pigment cells. In contrast, AxPG encodes a PG-M/versican-like proteoglycan that may promote the migration of A. mexicanum pigment cells, and AxPG expression is reduced in white mutant axolotls. We cloned a salamander orthologue of steel and used a partial genetic linkage map of Ambystoma to determine the genomic locations of steel, AxPG, and d. We show that the three genes map to different linkage groups, excluding steel and AxPG as candidates for d.

Ambystoma mexicanum↗

Ectopic anthocyanin pigmentation in maize as a tool for defining interactions between homologous regulatory factors.

The duplicated R and Sn genes are involved in the regulation of the maize anthocyanin biosynthetic pathway, encoding tissue-specific products that are homologous to the helix-loop-helix transcriptional activators. Sn determines the pigmentation of the mesocotyl, leaf basis and pericarp, while R determines pigmentation in various tissues, but not in the mesocotyl. In the progeny derived from test-crosses of R/Sn heterozygous plants, a high frequency of R plants exhibiting mesocotyl pigmentation was observed; these derivatives were defined as R*. In R* plants, the presence of this novel trait was not accompanied by the acquisition of Sn or by gross DNA rearrangements in the R profile. Accordingly, RT-PCR analysis showed that mesocotyl pigmentation in R* was attributable to the resident R gene. The occurrence of R* was observed with all R alleles tested, and was enhanced when a P component was present. The heritability of R* was shown only in the case of the standard R-r allele, which carries a functional P component. In addition, we observed that R* can influence other R alleles, transferring the ability to pigment the mesocotyl. R* is unstable, showing a tendency to return to its original state after a few generations. In R* plants there was a correlation between observed ectopic pigmentation and an increase in the level of A1 transcript but, surprisingly, not in the accumulation of R transcript. The results obtained from the analysis of test crosses of rSn/r delta plants suggest that an unlinked genetic factor accounts for the ectopic pigmentation. Concomitant occurrence of epigenetic events might explain the observed instability and reversibility noted above. Further study of this phenomenon might help to elucidate the basis of the interaction between homologous and non-homologous regulators.

Anthocyanins↗

Pigmented macular pucker.

A 66-year-old woman developed a darkly pigmented preretinal membrane. In the fundus periphery there was a round retinal hole about one disk diameter with pigmentation in its immediate vicinity. Pigmented, preretinal membranes arise from retinal pigment epithelium cells that exit through a retinal tear and settle on the inner retinal surface where they proliferate and form a membrane. Some preretinal membranes arise from unpigmented cells such as glial cells and become colored with iron pigment because of the accumulation of pigmented macrophages and free pigment debris. A retinal hole is not necessary for the occurrence of a glial membrane.

Aged↗

Formation of yellow, orange, and red pigments in the reaction of alk-2-enals with 2-thiobarbituric acid.

The reaction of four to eight carbon straight-chain alk-2-enals with 2-thiobarbituric acid (TBA) produced yellow 455-nm-, orange 495-nm-, and red 532-nm-absorbing pigments depending upon the reaction conditions. The 1:1 reaction of the aldehydes with TBA in 15% acetic acid at 100 degrees C produced the yellow pigment at 0.25 h and the red at 6 h. The reaction of the aldehydes with TBA in excess at 100 degrees C produced the yellow at 0.25 h, the orange at 2-6 h, and the red at 0.25-6 h. The formation of these pigments required molecular oxygen. These pigments could be separated from each other on HPLC. The red pigment formed from the aldehydes could not be distinguished from the red 1:2 malonaldehyde-TBA adduct by absorption spectrum and HPLC. The red color yield was the highest in the 1:1 reaction and retarded in the reaction with TBA in excess. The red color due to these aldehydes may contribute in part to the color formed in the general TBA test of lipid oxidation. The 1:1 reaction initially produced colorless 1:1 adducts X, which were subsequently converted into the yellow and red pigments under aerobic conditions. The reaction of the aldehydes with TBA in excess might initially produce X and then another colorless 1:2 adducts Y; the latter being converted into yellow, orange, and red pigments under aerobic conditions.

Aldehydes↗

Subcellular distribution of free and esterified forms of vitamin A in the pigment epithelium of the retina and in liver.

1. The distribution of vitamin A was examined in various subcellular fractions of rat liver and bovine retinal pigment epithelium. In rat liver, the major portion of the vitamin is in the cytosol, whereas in pigment epithelium, it is concentrated mainly in the microsomes. The microsomal vitamin A of pigment epithelium is tightly bound to membranes, as shown by the inability to release it except by organic solvent extraction or incubation with Triton X-100. 2. In both tissues, two different forms of cytosol vitamin A could be distinguished by ultracentrifugation. The major portion in liver is in the floating lipid phase and consists mainly of retinyl ester. The remainder (less than 10% of the total) is in the underlying infranatant; about 90% of the vitamin A in this fraction is esterified. By contrast, two-thirds of the vitamin A of pigment epithelial cell cytosol is in the infranatant; it consists of both esterified and unesterified retinol. The floating layer in the pigment epithelial cytosol consists entirely of retinyl ester. 3. These two forms of cytosol vitamin A in the pigment epithelium could also be separated by gel filtration on Sephadex G-100 which yielded two distinct fluorescent peaks. The first, which appeared in the void volume and corresponded in all probability to the floating layer obtained by ultracentrifugation, consisted only of retinyl ester. The second peak, which was eluted in approximately the same position as myoglobin, contained only unesterified retinol. It was abolished completely by preincubation with pronase. These findings support the view that the second peak represents the endogenous retinol-retinol binding protein complex of pigment epithelial cytosol. The fluorescent enhancement of the retinol bound to protein in this peak was about 4--5-fold compared to retinol in organic solvents.

Animals↗

Lens wound healing and cataractogenesis in a pigmented eye.

Light and electron microscopy and slitlamp microscopy were used to follow the development and partial repair of injury-induced cataract in the lens of the pigmented eye of the grey squirrel. These processes proceed in much the same way as in previously-studied albino rodents and rabbits in spite of invasion of the lens opacity by pigmented cells in the squirrel eye. Epithelial and capsular regeneration and lens fiber repair occur rapidly and apparently independently from the accumulation of pigmented cells, fibroblasts and collagen in the wound outside the lens epithelial layer. Using morphological criteria, some of the pigmented cells in the lens wound are identified as iris stromal melanocytes and pigmented epithelial cells; this is consonant with the slitlamp observations of streams of pigmented cells extending from the iris to the lens wound. The role, if any, of the pigmented cells in lens wound healing is unknown.

Animals↗

Spatial distribution of visual pigment and dopamine in the bullfrog retina.

Visual pigments and a neurotransmitter, dopamine, were quantitatively investigated in the retina of adult bullfrog, Rana catesbeiana. The adult bullfrog (body length 15-16 cm, body weight 375 +/- 52 g, n = 10) had 21.4 +/- 4.2 nmol visual pigment and 209 +/- 28 pmol dopamine in retinal areas of 266 +/- 27 mm2. Greater pigment densities were recorded in a semicircular band around the optic disc, extending to the nasal and temporal peripheries of the ventral retina. The area with the highest concentration of visual pigment was found in the middle of the dorsal retina, 3-4 mm dorsal to the optic disc. A high concentration of vitamin A2-based pigment was found in the dorsal quarter of the retina (porphyropsin zone); the zone extended up to the most ventral part along peripheral regions of the retina. There was also a band with higher dopamine concentrations although it was not so prominent as that of the visual pigment; the highest concentration of dopamine was found in the area immediately dorsotemporal to the optic disc. Fluorescence micrography indicated that the distribution pattern of catecholamine-containing amacrine cells paralleled that of the dopamine content. The topographic map of dopamine was slightly different from that of visual pigment in the bullfrog retina.

Animals↗

Retinal pigment epithelial cells produce PDGF-like proteins and secrete them into their media.

Human retinal pigment epithelial cells at confluence were used to condition serum-free Dulbecco's modified Eagle's medium. Conditioned media were exhaustively dialyzed against 0.5 N acetic acid, lyophilized, and subjected to Western blot analysis, using as primary antibody an IgG fraction prepared from goat antiserum directed against human platelet-derived growth factor. Native platelet-derived growth factor was resolved as a band with Mr of 30 kDa under non-reducing conditions, while bands with Mr of 36-38 kDa and 18.5 kDa were resolved from retinal pigment epithelial cell-conditioned media. Acid extracts of retinal pigment epithelial cells also contained bands at 36-38 kDa and media conditioned for 48 hr exhibited much denser bands than media conditioned for 24 hr. No bands were detected when non-immune goat IgG fractions were substituted for primary antibody and when conditioned media were prepared from several human fibroblast lines in the same manner as those prepared from retinal pigment epithelial cells, no detectable bands or only a faint shadow at 36 kDa were seen. Retinal pigment epithelial cell-conditioned media prepared in the presence of [35S]methionine were loaded on an anti-platelet-derived growth factor IgG affinity column, eluted, and subjected to SDS-polyacrylamide gel electrophoresis. Bands with Mr slightly less than 36 kDa and 18 kDa were visualized by autoradiography, demonstrating that the platelet-derived growth factor-like proteins in retinal pigment epithelial cell-conditioned media are newly synthesized. Two fractions eluted from the column also markedly stimulated fibroblast chemotaxis and incorporation of [3H]thymidine, both of which were neutralized by soluble anti-platelet-derived growth factor IgG. These data suggest that retinal pigment epithelial cells in culture produce platelet-derived growth factor-like proteins and secrete them into their media where they are capable of stimulating fibroblast chemotaxis and proliferation.

Cell Line↗

Physiologic pigmentation of the oral mucosa in Israeli children.

Physiologic melanin pigmentation (racial pigmentation) of the oral mucosa varies in prevalence among different races and ethnic groups. The purpose of this study was to investigate the prevalence of physiologic pigmentation in Israeli Jewish children of different ethnic origins. A total of 1,300 children, 6 to 10 years of age, was examined. Physiologic pigmentation was found in 13.5% of the population studied. Children of Eastern origin showed a significantly higher prevalence of pigmentation compared with Ashkenazi and Sephardic groups. Because melanin pigmentation can be enhanced by mechanical and chemical stimulation (smoking), this study may serve as a baseline for investigation of melanin pigmentation in various ethnic groups.

Child↗

Macular pigment and reduced foveal short-wavelength sensitivity in retinitis pigmentosa.

Some patients with retinitis pigmentosa (RP) show a reduced foveal short-wavelength sensitivity that cannot be attributed to a reduction in the sensitivity of the short-wavelength cone system. To determine whether an increased amount of macular pigment (xanthophyll) might account for this finding, we derived estimates of the two-way optical density of the macular pigment of five such RP patients as well as of five normals. The spectral reflectance of the foveal region of each subject was obtained from digitized images of the bleached fundus provided by a television-based reflectometer. The density spectra of the macular pigment, melanin, and oxygenated hemoglobin were fit by a least-squares procedure to the log of the ratio of parafoveal to foveal spectral reflectance in order to obtain a quantitative estimate of the contribution of each of these ocular pigments to foveal short-wavelength reflectance. By this analysis, the two-way densities of the macular pigment, melanin, and oxyhemoglobin of the RP patients were not significantly different from those of the normals. Therefore, the reduced foveal short-wavelength sensitivity of these patients was not due to an increased amount of macular pigment, but may result instead from morphological abnormalities in the foveal cones such that a normal amount of macular pigment screens the cones more effectively.

Adult↗

An attempt to control the polychlorocatechol pigment production during 3-chlorobenzoate aerobic co-metabolism in growing-cell batch culture.

A strain of Pseudomonas fluorescens, designed CP30, was capable of co-metabolizing 3-chlorobenzoic acid via chlorocatechols, thus producing a polychlorocatechol black pigment. The pigment production was found to be enzyme-mediated in P. fluorescens cultures; however, the results of the autoxidation studies suggested that a large amount of pigment could derive from the simple autoxidation of the chlorocatechols accumulated in the CP30 culture during the 3-chlorobenzoate degradation. The polymerization of chlorocatechols should reduce the toxicity of the culture and generally resulted in a large production of a water-insoluble (i.e. a easily removable) pigment with the release a large amount of chloride ions. Both 3-chlorobenzoate degradation and pigment production were stimulated and inhibited in the same way by high and low pH values, respectively, in CP30 cultures; the culture conditions which permitted an extensive 3-chlorobenzoate degradation with the lowest pigment production were not identified. However, on the basis of the detoxification property of the pigment production process, high pH and dissolved oxygen concentrations appears to be the most suitable culture conditions for an enhanced detoxicant treatment of 3-clorobenzoate-contaminated waste water.

Aerobiosis↗

The control of pigment migration in isolated erythrophores of Holocentrus ascensionis (Osbeck). II. The role of calcium.

The integumental pigment cells (erythrophores) of the squirrel fish, Holocentrus ascensionis, are specialized for rapid radial transport of the pigment granules contained within their cytoplasm. Pigment granules in isolated denervated erythrophores alternate spontaneously between a centrally aggregated state and a radially dispersed state. In the absence of external calcium, pigment aggregation does not occur spontaneously and cannot be induced by the aggregating agents epinephrine or high concentration of external K+. Pigment aggregation is also impaired in the presence of D600 or papaverine, compounds reported to antagonize calcium influx into the cell. Pigment aggregation can be induced by experimental elevation of the concentration of cytoplasmic free Ca2+, with a Ca-EGTA buffer system in conjunction with ionophore A23187. The threshold concentration of Ca2+ required to produce this effect is 5 X 10(-6) M. These results suggest that cytoplasmic free Ca2+ is involved in mediating pigment aggregation and that some, if not all, the Ca2+ is supplied by influx from the extracellular space.

Animals↗

Spectral properties of a mixture of fluorescent pigments produced by Pseudomonas aeruginosa.

The major chromophore of a mixture of fluorescent pigments produced by Pseudomonas aeruginosa ATCC 9027 had pH-dependent absorption, excitation, and emission spectra, such that two ionic forms existed in the ground state and three in the excited states. The pigments could complex with several metal ions to change fluorescence and absorption spectra. Although the pigments were separable into several components, spectra indicated that the same fluorescent chromophore was present in each component. Hydrolysis of the mixture of pigments gave amino acids which did not include alanine or lysine. These pigments must therefore differ from those described by other workers, even though similarities of the chromophores were evident from comparisons with data in the literature, and from comparisons of a hydrolytic product of the mixture of pigments, termed compound F, with the chromophore of the fluorescent pigment of Azotobacter vinelandii. Drastic hydrolysis of the latter chromophore also yielded compound F.

Azotobacter↗