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Albino and pigmented gerbil auditory function: influence of genotype and gentamicin.

Auditory function of albino and pigmented gerbils was examined before and after treatment with the ototoxic aminoglycoside antibiotic gentamicin. Auditory brainstem responses (ABRs) and cochlear nerve compound action potentials (CAPs) were measured in response to pure tones having frequencies between 2 and 32 kHz. Age-matched albinos had significantly lower CAP, but not ABR, thresholds than pigmented gerbils. Gentamicin treatment elevated CAP and ABR thresholds in both genotypes, but pigmented gerbils were less severely affected. Compared to the ABR, the CAP is a more sensitive measure of ototoxicity and pigmentation differences. CAP tuning curves (TCs) were another sensitive measure of genotypic differences in susceptibility to ototoxicity. TC tip thresholds from pigmented animals given gentamicin were not as elevated as the TC tip thresholds of albinos.

Animals↗

An improved method for isolation and culture of pigment epithelial cells from rat retina.

An improved method for isolation and culture of pigment epithelial cells from normal rat retinas is described. Following a brief incubation in the neutral protease Dispase, large epithelial sheets can be harvested rapidly. The separation of the choroid from the pigment epithelium prior to retinal detachment greatly reduces the risk of contamination of the cultures with choroidal cells. Growth of pigment epithelial cells on Matrigel-coated microporous filters in hormonally-defined medium allows for development of high levels of transepithelial electrical resistance as well as for preservation of the differentiated pigment epithelial cell phenotype. This method should be useful for studies of pigment epithelial cell permeability and structural differentiation in vitro.

Animals↗

Extracellular matrix mediated growth and differentiation in human pigment epithelial cell line 0041.

Efforts to grow differentiated pigment epithelial cells have led to a characterization of the growth kinetics of spontaneously established, continuously growing, human retinal pigment epithelial (PE) cell line 0041 on several biomatrices. These substrates were prepared from (a) placental and amniotic membrane, (b) commercially available basement membrane matrix (Matrigel), (c) dishes coated with extracellular matrix secreted by endothelial cells (ECM), (d) dishes coated with collagen IV and/or laminin, (e) dishes coated with collagen I and/or fibronectin. Our findings suggest that tissue culture plastic and dishes coated with collagen IV alone promote higher cell densities, while highest plating efficiency (24 hrs) was seen on tissue culture plastic and Matrigel. The highest degree of differentiation (epithelioid appearance, apical villi and junctional complexes) was seen in cells plated on dishes coated with collagen IV and extracellular matrix secreted by endothelial cells. Cells were epithelioid and polarized on those two substrates; they expressed fine finger-shaped villi and the highest degree of cell contact (in the form of junctions). Cells grown on Matrigel looked like fibroblasts and became deeply pigmented; however, the nature of the pigment remains to be determined. Collagen IV and ECM coated dishes, therefore, are most suitable for cultures of human PE cell line 0041 because they provide higher cell densities while retaining the differentiated state. This is the first report where an established pigmented epithelial cell line has been induced to become differentiated by use of extracellular matrices and extracellular matrix components.

Adolescent↗

Uveitogenic 28/30 kD and 43 kD polypeptides in pigment epithelial membranes of the retina.

UNLABELLED: The purpose of this study was the search for new intrinsic polypeptides of the retinal pigment epithelium (RPE) capable of evoking experimental uveitis. A membrane fraction was prepared from purified bovine RPE cells. The Triton X-100 soluble protein fraction was separated into polypeptide fractions by preparative gel electrophoresis, and the pathogenicity of the main isolated polypeptides was investigated in Lewis rats. After immunization, two hitherto unknown pigment epithelial polypeptides with M(r) 28/30 kD (doublet) and 43 kD (PEP-28/30 and PEP-43, respectively) elicited progressive pigment epitheliitis and choroiditis accompanied by extending plaque-shaped macrophage accumulations along the RPE-Bruch's membrane layer. No inflammatory foci were found within the neural retina. Polypeptide fractions with M(r) 14-17, 25 and 32/34 kD (doublet) (PEP-14/17, PEP-25 and PEP-32/34, respectively) appeared to be non-uveitogenic at the tested dose. PEP-28/30 and PEP-43 exhibited a partial antigenic relationship to PEP-65. PEP-28/30 exhibited marked reactivity to a monoclonal antibody previously raised to a 32 kD RPE-specific protein. IN CONCLUSION: in addition to the previously described main RPE-specific membrane polypeptide PEP-65, the RPE appears to contain two more uveitogenic components, the intrinsic pigment epithelial membrane polypeptides PEP-28/30 and PEP-43. Like PEP-65, these antigens are able to evoke experimental autoimmune pigment epithelial protein-induced uveitis (EAPU) in rats.

Animals↗

Acid hematin pigmentation of the cornea in stillborn fetuses.

During the postmortem histopathologic evaluation of eyes from stillborn fetuses we noted the presence of a prominent undescribed corneal pigment in 18 of 55 stillborn fetuses. The corneal pigment was frequently associated with documented meconium-stained amniotic fluid, and in no instance was a stained cornea coupled with recorded clear amniotic fluid. Pigmented corneas came from stillborn fetuses with a longer duration of intrauterine death than nonstained corneas. The pigment stained black with the Fontana-Masson stain, was birefringent, and treatment of tissue sections with 5% potassium permanganate and 5% oxalic acid as well as with saturated alcoholic picric acid solution removed the pigment indicating that it is acid hematin. The most likely cause of the acid hematin-stained corneas was tissue acidity created in utero with prolonged intrauterine death.

Cornea↗

Melanosis coli--histochemical and immunohistochemical comparison of the pigments of melanosis coli and Dubin-Johnson syndrome.

We compared the pigment of melanosis coli with the pigment of Dubin-Johnson syndrome, melanin, and lipofuscin. The pigment of melanosis coli appeared similar to lipofuscin in that it stained positively with periodic acid-Schiff, oil red-0 and Victoria blue stains and revealed negative reactions to the immunohistochemical stains for S-100 protein and neuron specific enolase, but had similarity to melanin as shown by the positive reaction to Fontana-Masson stain and negative autofluorescence. The pigment of Dubin-Johnson syndrome showed the same histochemical and immunohistochemical characteristics as that of melanosis coli. The results indicate that the pigments of melanosis coli and Dubin-Johnson syndrome are identical and are variants of lipofuscin.

Colonic Diseases↗

The influence of the light transmission properties of plastic window coverings on broiler pigmentation.

An experiment was conducted to determine the effect of different window coverings on broiler pigmentation. Equal numbers of male and female broilers were grown in open-type houses. The control house had open sides, while each treatment house had window coverings with either 42% or 89% light transmission properties. Birds from houses with window coverings deposited the same amount of pigment in the skin as did birds from the open house as indicated by blood xanthophyll level and excitation purity. Skin color scores, however, as well as dominant wavelengths and luminosities of shanks from these birds, indicated that pigmentation of birds grown in the house with 42% light transmission window coverings was significantly inferior to that observed for birds grown in either the open house or the house with 89% light transmission window covering. It was concluded that in situations where pigmentation is of concern, care should be taken in selecting window coverings which will not adversely affect pigmentation.

Animals↗

The effects of sex steroid hormones on the pigment accumulation in the shell gland of Japanese quail.

Effects of sex steroid hormones on the accumulation of the superficial pigment of the eggshell in the quail shell gland were investigated. In Experiment 1, 10 or 20 mg/100 g BW of aminoglutethimide (AG) was injected s.c. into laying quail hens 10 h before the expected ovulation. Thirty percent of the hens that received 10 mg/100 g BW and 87% of the hens that received 20 mg/100 g BW failed to ovulate. They had significantly (P < .01) lower concentrations of preovulatory plasma progesterone (P) than the control hens and showed significantly (P < .05) low concentrations of plasma estradiol-17 beta (E) and small amounts of the pigment in their shell glands after oviposition of the preceding egg. In Experiment 2, AG (20 mg/100 g BW) was administered s.c. to quail hens 10 h before the expected ovulation, followed by the administration of P (.05, .1, or .2 mg/100 g BW), E (.1, .2, or 1.0 mg/100 g BW), or testosterone (T) (.5 mg/100 g BW). Significant increases in the accumulated pigment 18 h after oviposition of the preceding egg were found when P was injected. However, administration of E and T had no influence on pigment accumulation. These results suggested that P might affect the accumulation of pigment in the quail shell gland.

Animals↗

Macular hole with retinal pigment epithelium hyperplasia simulating neoplasm.

A case in which a retinal pigment epithelium migration through a stage IV macular hole onto the retinal surface simulated a retinal pigment epithelium neoplasm is described. A 69-year-old woman presented with a hyperpigmented membranous retinal surface growth temporal to a stage IV macular hole in the left eye. Clinical examination and optical coherence tomography were performed prior to pars plana vitrectomy with membrane peeling. Histopathologic processing revealed melanosomes and retinal pigment epithelium with no evidence of neoplastic cells. Retinal pigment epithelium migration can occur through a full-thickness macular hole onto the retinal surface, simulating a retinal pigment epithelium neoplasm. It may be a poor prognostic indicator for visual success.

Aged↗

Genetics of pigmentation in Biomphalaria straminea.

A monogenetic factor determining basic pigmentation in Biomphalaria straminea follows simple. Mendelian inheritance and has at least three alleles, as in B. glabrata. Wild type pigmentation with black body, mantle collar and eyes is dominant; albino with no black pigment is recessive; and "blackeye" pigmentation, deficient in black pigment in body and mantle collar, is recessive to wild type but dominant over albino.

Alleles↗

Factors influencing the number needed to excise: excision rates of pigmented lesions by general practitioners.

OBJECTIVE: To identify doctor and patient characteristics associated with excision of benign versus malignant pigmented skin lesions. DESIGN, SETTING AND PARTICIPANTS: Retrospective audit of data on 4741 pigmented skin lesions excised from November 1998 to February 2000 by 468 general practitioners (39% response rate) from 223 practices in Perth, WA. (The data used were from the baseline period of a randomised controlled trial of a diagnostic aid for pigmented skin lesions.) MAIN OUTCOME MEASURE: The number needed to treat (NNT), defined as the number of pigmented lesions needed to be excised to identify one melanoma, in relation to demographic characteristics of GPs and patients. RESULTS: Relatively more benign lesions were excised per melanoma (NNT = 83) in the youngest patients (aged 10-19 years) compared with the oldest (aged > or = 70) (NNT = 11) (P [trend] < 0.001), in females (NNT = 37) compared with males (NNT = 23) (P = 0.02), and in the most socioeconomically disadvantaged (NNT = 60) compared with the least disadvantaged group (NNT = 20) (P [trend] < 0.001). The most recently graduated GPs excised more benign lesions for each melanoma (NNT = 59) than the least recently graduated (NNT = 22) (P [trend] = 0.01). CONCLUSIONS: GPs could raise their threshold for excising pigmented lesions in patients who are young, female, or from areas of low socioeconomic status, or if the GPs themselves are recent graduates.

Adolescent↗

Overexpression of a gene cluster encoding a chalcone synthase-like protein confers redbrown pigment production in Streptomyces griseus.

A 7.0-kb DNA fragment that conferred redbrown pigment production on Streptomyces griseus was shotgun-cloned with a multicopy vector pIJ486 from this microorganism. By restriction endonuclease mapping and subcloning, a 1.5-kb fragment which is essential for the production of redbrown pigment was determined. The nucleotide sequence of this region revealed the presence of two open reading frames, ORF1 with 109 amino acids (named RppA) and ORF2 with 262 amino acids (RppB), in addition to a truncated ORF3. The termination codon of rppA and the initiation codon of rppB overlapped, sharing one common nucleotide, which strongly suggests that these two genes are cotranscribed. Both rppA and rppB were essentially required for the pigmentation. The RppB protein showed great similarity in amino acid sequence to a chalcone synthase, a key enzyme of central importance in the biosynthetic pathway of all classes of flavonoids in plants. Part of RppA showed sequence similarity to the 33kDa phosphoprotein of adenovirus. Nucleotide sequences homologous to rppA and rppB were widely distributed in Streptomyces species, as determined by Southern hybridization. Further nucleotide sequencing of the entire orf-3 gene showed that ORF3 with 403 amino acids was a cytochrome P-450 (named P-450RPP). These data suggested that the cloned fragment contained part of a gene cluster for the biosynthesis of a certain metabolite. Introduction of the subcloned 1.5-kb fragment into Streptomyces lividans as well as Escherichia coli also caused production of redbrown pigment, suggesting that RppA and RppB are capable of synthesizing the redbrown pigment from metabolites commonly present in bacteria.

Acyltransferases↗

Structures of grixazone A and B, A-factor-dependent yellow pigments produced under phosphate depletion by Streptomyces griseus.

A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) acts as a microbial hormone that induces morphological development and secondary metabolism in Streptomyces griseus. A diffusible yellow pigment is produced by S. griseus in an A-factor-dependent manner under phosphate depletion. Detailed analysis of the pigment production by S. griseus cultivated in minimal liquid medium containing different concentrations of phosphate showed that the pigment was actively produced in the presence of low concentrations of phosphate and the production of the pigment was completely repressed in the presence of 2.5 mM KH2PO4. HPLC analysis of the culture supernatant showed that the pigment consisted of two major, structurally related compounds and they were produced at different ratios depending on the concentration of phosphate in the medium. The structures of the two major compounds, designated as grixazone A and B, were determined by spectroscopic analyses as 1-[[2-(acetylamino)-2-carboxyethyl]thio]-2-amino-3-oxo-8-formyl-3H-phenoxiazine and 1-[[2-(acetylamino)-2-carboxyethyl]thio]-2-amino-3-oxo-8-carboxyl-3H-phenoxiazine, respectively. Grixazone A was a novel compound, although grixazone B was reported in a patent as a parasiticide produced by Streptomyces sp. DSM3813.

Chromatography, High Pressure Liquid↗

[An epidemiologic examination on the prevalence of the periodontal diseases and oral pigmentation in Yusho patients in 1998].

An epidemiologic examination was carried out to reveal the prevalence of the periodontal diseases and oral pigmentation in patients with Yusho 30 years after PCBs exposure. The results obtained were as follows. 1) 69 patients out of 71 patients with Yusho, who were measured periodontal pocket depth using Ramfjord' methods, had at least one tooth with periodontal pocket deeper than 3 mm. Similarly, 241 teeth out of a total 348 examined teeth showed periodontal pocket with more than 3 mm depth. 2) Oral pigmentation was observed in 46 out of 79 patients with Yusho. In this study, gingival pigmentation was most predominant among oral pigmentation. In addition, it is of particular interest that oral pigmentation tended to be observed at a much higher frequency in younger patients with Yusho.

Adult↗

Color tuning mechanism of human red and green visual pigments.

We investigated the molecular mechanism of a rather large red shift of 31 nm in a human red pigment compared with a human green pigment. In this analysis, we paid special attention to the phenomenon of nonadditivity of spectral shifts due to substitution of the key amino acids (OH-bearing amino acids) and the phenomenon of cooperativity by which the spectral shifts due to substitution of the key amino acids in the protein environment of red pigment are about 1.5 times larger than that in the protein environment of green pigment. The analysis was made by using a model of three active sites on which the key amino acids are located and four effective sites by which the effect of the key amino acids is modified. As a result, we found that the interaction between the active sites that occurs through the repolarization of the chromophore induced by the key amino acid is essential for the nonadditivity phenomenon. We also found that the interaction between the active site and the effective site plays a major role in the cooperativity phenomenon. More directly, we say that the highly polarizable property of the chromophore is the origin of the rather large red shift in red pigment. Based on these analyses, we conclude that the interaction between the polarizable chromophore and the protein moiety has the capability of producing a significant spectral shift, at least 1000 cm-1, even by substitution of moderate polar residues of the OH-bearing amino acids.

Binding Sites↗

Blue- and green-absorbing visual pigments of Drosophila: ectopic expression and physiological characterization of the R8 photoreceptor cell-specific Rh5 and Rh6 rhodopsins.

Color discrimination requires the input of different photoreceptor cells that are sensitive to different wavelengths of light. The Drosophila visual system contains multiple classes of photoreceptor cells that differ in anatomical location, synaptic connections, and spectral sensitivity. The Rh5 and Rh6 opsins are expressed in nonoverlapping sets of R8 cells and are the only Drosophila visual pigments that remain uncharacterized. In this study, we ectopically expressed Rh5 and Rh6 in the major class of photoreceptor cells (R1-R6) and show them to be biologically active in their new environment. The expression of either Rh5 or Rh6 in "blind" ninaE(17) mutant flies, which lack the gene encoding the visual pigment of the R1-R6 cells, fully rescues the light response. Electrophysiological analysis showed that the maximal spectral sensitivity of the R1-R6 cells is shifted to 437 or 508 nm when Rh5 or Rh6, respectively, is expressed in these cells. These spectral sensitivities are in excellent agreement with intracellular recordings of the R8p and R8y cells measured in Calliphora and Musca. Spectrophotometric analyses of Rh5 and Rh6 in vivo by microspectrophotometry, and of detergent-extracted pigments in vitro, showed that Rh5 is reversibly photoconverted to a stable metarhodopsin (lambda(max) = 494 nm), whereas Rh6 appears to be photoconverted to a metarhodopsin (lambda(max) = 468 nm) that is less thermally stable. Phylogenetically, Rh5 belongs to a group of short-wavelength-absorbing invertebrate visual pigments, whereas Rh6 is related to a group of long-wavelength-absorbing pigments and is the first member of this class to be functionally characterized.

Amino Acid Sequence↗

Molecular motors and their role in pigmentation.

Skin pigmentation is orchestrated through a series of complementary processes. After migration of melanoblasts out of the neural crest to epidermis and hair follicle, these cells mature into melanocytes. Differentiated melanocytes produce melanin in specialized organelles, the melanosomes. Moreover, the cytoplasm of melanocytes branches into extensions, the dendrites. Via the tips of these dendrites they donate their mature melanosomes to the keratinocytes resulting in skin pigmentation. Thus, one essential part of the process of pigmentation is the translocation of melanosomes from their site of origin in the perinuclear cytoplasm towards the dendrite tips. Motor proteins are molecules which use the energy derived from ATP hydrolysis to move along cytoskeletal elements, either actin filaments or microtubules, to transport their cargo, which can be organelles, vesicles or chromosomes. This review describes the different classes of microtubule-based and actin-based motor proteins with their characteristics and functional importance in cell biology and organelle transport. Some of them will be highlighted and several recent studies in mammalian pigment cells indicating their role in pigment granule transport will be discussed. As a result of these data and previous suggestions, a model will be proposed for the possible cooperation of both systems in melanosome movement.

Actin Cytoskeleton↗

Clinicopathological correlation of pigmented skin lesions using dermoscopy.

Dermoscopy (dermatoscopy, epiluminescence microscopy) is an additional measure for making the diagnosis of pigmented skin lesions more accurate. It enables the clinician to visualize features not discernible by the naked eye. By applying enhanced digital dermoscopy and a standardized gross pathology protocol to pigmented skin lesions, a precise clinicopathological correlation of relevant dermoscopic features can be made. Histological specimens of four pigmented skin lesions (melanoma in situ, Clark's nevus, Reed's nevus, seborrheic keratosis) were processed using a standardized gross pathology protocol and viewed along with the clinical photographs and digital dermoscopic images that were magnified and enhanced to better visualize the corresponding dermoscopic structures. Furthermore, measurements of dermoscopic structures using digital equipment were correlated with histometric findings. Our understanding of dermoscopic features, especially the broadened pigment network - a specific dermoscopic criterion for melanoma - was refined by this detailed case-by-case correlation. In addition, some not yet fully characterized dermoscopic features, such as black lamella, radial streaks, and exophytic papillary structures, were described in detail dermoscopically and histopathologically. Moreover, measurements of these dermoscopic features and the underlying histological structures were found to be similar. Linking dermoscopy more closely with cutaneous pathology may help refine the definitions and diagnostic criteria of pigmented skin lesions for dermatologists as well as dermatopathologists.

Adult↗