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Evidence for several roles of dynein in pigment transport in melanophores.

Melanophores are specialized cells that transport pigment granules to and from the cell center, giving animals the ability to change skin color. A kinesin-related plus-end motor has previously been shown to be responsible for pigment granule dispersion [V.I. Rodionov, F.K. Gyoeva, and V.I. Gelfand. Proc. Natl. Acad. Sci. USA. 1991, 88:4956-4960]. Here, we have microinjected a dynein antibody (70.1) into cultured cod (Gadus morhua) melanophores and used the dynein inhibitor vanadate on permeabilized melanophores in skin pieces, to examine the role of the microtubule minus-end motor dynein in these cells. Both pigment granule aggregation and maintenance of the spherical central pigment mass (CPM) were inhibited by the antibody and by vanadate. Vanadate or antibody treatment of cells with aggregated pigment did not induce pigment dispersion. However, when the antibody-injected cells were induced to disperse pigment, the pigment moved farther to the cell periphery, which resulted in a depletion of pigment in the cell center. Similar superdispersion of previously uniformly distributed pigment was also seen when the antibody was injected in melanophores with dispersed pigment. Our results demonstrate that both pigment aggregation and maintenance of the CPM are dynein-dependent processes. Our data further show that dynein is involved in the homogeneous distribution of dispersed pigment. These results suggest that both dynein and kinesin are active in keeping pigment granules dispersed throughout the cytoplasm, transporting pigment granules in opposite directions. The possibility that dynein is continuously active during both aggregation and dispersion, while kinesin might be the target for regulation, is discussed.

Animals↗

Ultrastructure and migration of screening pigments in the retina of Pieris rapae L. (Lepidoptera, Pieridae).

The retinal morphology of the butterfly, Pieris rapae L., was investigated using light and electron microscopy with special emphasis on the morphology and distribution of its screening pigments. Pigment migration in pigment- and retinula cells was analysed after light-dark adaptation and after different selective chromatic adaptations. The primary pigment cells with white- to yellow-green pigments symmetrically surround the cone process and the distal half of the crystalline cone, whilst the six secondary pigment cells, around each ommatidium, contain dark brown pigment granules. The nine retinula cells in one ommatidium can be categorised into four types. Receptor cells 1-4, which have microvilli in the distal half of the ommatidium only, contain numerous dark brown pigment granules. On the basis of the pigment content and morphology of their pigment granules, two groups of cells, cells 1, 2 and cells 3, 4 can be distinguished. The four diagonally arranged cells (5-8), with rhabdomeric structures and pigments in the proximal half of the cells, contain small red pigment granules of irregular shape. The ninth cell, which has only a small number of microvilli, lacks pigment. Chromatic adaptation experiments in which the location of retinula cell pigment granules was used as a criterium reveal two UV-receptors (cells 1 and 2), two green receptors (cells 3 and 4) and four cells (5-8) containing the red screening pigment, with a yellow-green sensitivity.

Adaptation, Physiological↗

Skin responses to ultraviolet radiation: effects of constitutive pigmentation, sex, and ancestry.

Constitutive skin pigmentation and skin responses to ultraviolet radiation were measured on a sample of volunteers (n=250) living in State College, PA, USA. The sample was composed of individuals of European American (n=190), Hispanic (n=45), and East Asian ancestry (n=15). Constitutive pigmentation was measured using the Adjusted Melanin Index (AMI), Erythemal Dose Response (EDR) was measured using the slope of a* at 24 h (Deltaa*), and Melanogenic Dose-Response (MDR) was measured using DeltaAM, the slope of AMI at 7 d. The relationships between constitutive skin pigmentation, EDR, MDR, sex, age, and ancestry were investigated. European Americans showed a lower constitutive pigmentation, had a significantly higher burn response (EDR), and had a significantly lower tanning response (MDR) than Hispanics and East Asians. No significant difference is seen between Hispanics and East Asians for either constitutive pigmentation or EDR. Constitutive pigmentation in females was slightly lower than in males in all three samples, but the difference was not significant. While no differences were observed in MDR between sexes, males had a stronger EDR than females regardless of population or constitutive pigmentation level, and this difference was significant in European Americans and Hispanics. We observed no age-related differences in any of the populations or measures investigated. We evaluated the relationship between constitutive pigmentation, EDR and MDR. There was a strong inverse correlation between constitutive pigmentation and EDR in the three samples (European Americans, R2=0.176, P < 0.001; Hispanics, R2=0.204, P=0.009; East Asians, R2=0.223, P=0.098) and a strong direct correlation between constitutive pigmentation and MDR in European Americans and Hispanics (European Americans, R2=0.094, P < 0.001; Hispanics, R2=0.164, P=0.012). In other words, persons with lower constitutive pigmentation both burn more and tan less than persons with higher pigmentation. However, after controlling for constitutive pigmentation, EDR and MDR were significantly correlated in European Americans (R2=0.041 P=0.006). Thus, the general observation that persons who burn more tan less is probable because of the common link that these two phenotypes have with constitutive skin pigmentation and, in fact, once pigmentation has been adjusted for, there is a positive correlation between tanning response and burning response in European Americans.

Adolescent↗

Incidence and severity of iris pigmentation on latanoprost-treated glaucoma eyes.

PURPOSE: The purpose of the present study was to investigate the incidence and severity of iridial pigmentation under latanoprost topical use on brown eyes in Taiwan. METHODS: Retrospective review study was conducted from April 1999 to October 2001 in the Department of Ophthalmology, Taipei Veterans General Hospital, Taiwan, for glaucoma clinic monthly follow-up patients; 140 open-angle glaucoma patients on 0.005% latanoprost were enrolled. Analyses of iridial pigmentation incidence, grading, patient age distribution, side effect, and time course were performed. Boys-Smith pigment gradation lens was used as standard for semiquantitative iris pigmentation grading. RESULT: Before 0.005% latanoprost use, 90% of the patients enrolled were noted with iridial pigmentation grade I, and 10% were with grade I-II, but not reaching grade II scale standard. A total of 60 patients on 0.005% latanoprost developed increased pigmentation of the iris during the follow-up period. An increase of iris pigmentation was noted after an average of 7.27 months use of latanoprost (range 1-19 months, SD 2.65 months). For iridial pigmentation grading, 57.1, 30.7, 10.0, and 2.1% of our patients were noted to have grade I, II, III, and IV respectively. Most patients with latanoprost-induced iris hyperpigmentation were with grade II iridial pigmentation. There were 15 patients (10.7%) (10 female and five male) with hypertrichosis in the study group who were not compatible with the iridial pigmentation status. Among these patients, female patients had higher incidence of hypertrichosis than males, but this did not bother them. Only four patients (2.8%) were with conjunctiva chemosis and three patients (2.1%) with lid margin hyperpigmentation. CONCLUSION: Contrary to the belief that latanoprost rarely caused iris hyperpigmentation in yellow-brown eyes, our study showed that 42.8% iris hyperpigmentation did occur, especially after continual use for around 7 months. Higher hyperpigmentation incidence were noted in male than in female patients. This might be due to stronger adrenergic incidence in male than in female patients. Although hypertrichosis and increasing eyelid pigmentation together with iridial pigmentation represented a potentially permanent cosmetic side effect, they are very rare and occurred in no more than 3% in our patients. It is a good way to take Boys-Smith pigment gradation lens for iridial pigmentation grading and for long-term continual evaluation. The doctors should exert great care in differentiating drug-induced iris pigmentation and iris nevi from early stage uveal melanoma.

Adolescent↗

Short-term study of retinal pigment epithelium sheet transplants onto Bruch's membrane.

The purpose of this study is to investigate the survival and behaviour of retinal pigment epithelium sheets transplanted onto hydraulically debrided Bruch's membrane. Uncultured retinal pigment epithelium sheets obtained from male cats and sandwiched between two gelatin sheets were transplanted onto the tapetal area of female cats after native retinal pigment epithelium was debrided. For controls, the gelatin carrier was transplanted after debridement. Each transplant or control specimen was analyzed histologically and immunohistochemically. Transplanted male retinal pigment epithelial cells were identified by in situ labelling of the cat Y chromosome. Over half of the transplants appeared as retinal pigment epithelium multilayers in the subretinal space. Retinal pigment epithelium pigment dispersion into the subretinal space was seen in most of the transplants, and retinal pigment epithelium pigment infiltration into the neural retina was seen in all 7-day survival transplants. A few condensed darkly stained retinal pigment epithelium nuclei and Terminal Transferase dUTP Nick End Labelling-positive retinal pigment epithelium cells were observed in all transplants. Cellular retinaldehyde-binding protein was present up to day-7 in most transplanted RPE cells. In both transplant and control specimens, the antibody against the Ki-67 nuclear antigen labelled a few retinal pigment epithelium cells at day-3. Terminal Transferase dUTP Nick End Labelling-positive outer nuclear layer nuclei were most frequently observed at day-1 but were much less frequent at day-3 in both transplants and controls. The survival and effectiveness of retinal pigment epithelium sheet transplants appeared similar to the retinal pigment epithelium microaggregates transplants conducted previously in this model.

Animals↗

Indocyanine green videoangiographic findings in detachment of the retinal pigment epithelium.

BACKGROUND: Several forms of retinal pigment epithelial detachment have been reported. The authors used indocyanine green (ICG) videoangiography, which is useful to study the choroidal vasculature and Bruch membrane, to study pigment epithelial detachments. METHODS: Ninety-eight pigment epithelial detachments in 75 eyes were classified based on the appearance of choroidal neovascular membranes or late phase findings of ICG videoangiography done at the initial examination. The authors also followed the evolution of 51 such detachments not associated with choroidal neovascularization (CNV). RESULTS: Sixty-four pigment epithelial detachments without CNV were divided into five groups. Among eyes with pigment epithelial detachments that showed intense hyperfluorescence, all except one of the patients had both eyes involved and had several pigment epithelial detachments, sometimes with exudative retinal detachments. Weak hyperfluorescence was observed more often in younger patients. During follow-up of eyes with pigment epithelial detachments that showed irregular hypofluorescence, a neovascular membrane developed in one eye, microrips developed in four eyes and retinochoroidal folds in one eye. Most eyes that showed irregular hyperfluorescence developed atrophy of the retinal pigment epithelium. In 34 pigment epithelial detachments with CNV, either irregular hypofluorescence or absence of fluorescence was observed in areas that corresponded to the pigment epithelial detachment. CONCLUSION: The intense hyperfluorescence is thought to be due to the accumulation of protein-rich fluid within the pigment epithelial detachment. Most pigment epithelial detachments that showed weak fluorescence probably represent variants of central serous choroidopathy. Pigment epithelial detachments that showed irregular hypofluorescence or hyperfluorescence were associated with age-related macular degeneration, and the former was correlated closely with CNV. Close follow-up therefore is recommended for eyes with pigment epithelial detachments that show irregular hypofluorescence.

Adult↗

Pigmented viral warts: a clinical and histopathological study including human papillomavirus typing.

Although clinical, histological and viral correlations have recently been established among pigmented warts, homogeneous intracytoplasmic inclusion bodies and related types of human papillomavirus (HPV) (HPV 65, 4 and 60), the causes of the pigmentation remain unknown. In this study, comparative histological and histochemical analyses were performed with 53 pigmented (34 HPV 65-induced, 12 HPV 4-induced and seven HPV 60-induced) and 73 non-pigmented warts (27 HPV 2-induced, 23 HPV 1-induced, 12 HPV 63-induced, six unknown HPV-type induced and five HPV 60 induced) to clarify the causes of the pigmentation. Electron microscopy was also used to examine the pigmented warts. Many melanin blockade melanocytes were identified in all of the pigmented warts with Masson-Fontana staining and electron microscopy, and increased melanin in keratinocytes was also noted in 22 pigmented warts, suggesting that the dispersion of melanin granules in the dendrites of the melanin blockade melanocytes and the increased melanin granules in keratinocytes are the primary contributors to the pigmentation of the warts. The homogeneous intracytoplasmic inclusion bodies might also play a part in the darkening of the warts, as only the cases which had the inclusion bodies as well as the melanin blockade melanocytes were clinically pigmented. Although melanin blockade melanocytes were seen in a few cases of HPV 1- and HPV 2-induced warts in which the homogeneous inclusion bodies were not observed, the warts were not clinically pigmented. Melanin blockade melanocytes were not seen in any of the HPV 63-induced non-pigmented warts. In conclusion, the pigmented warts were associated with one of the related types of HPV (HPV 65, 4 and 60), and the pigmentation of the lesions is thus thought to be caused primarily by melanin blockade melanocytes. The homogeneous intracytoplasmic inclusion bodies might also play a part in the darkening of the lesions. This is the first report dealing with the pigmentary disorder associated with specific types of HPV.

Adolescent↗

Impairment of macrophage functions after ingestion of Plasmodium falciparum-infected erythrocytes or isolated malarial pigment.

Human monocyte-derived macrophages ingest diamide-treated red blood cells (RBC), anti-D immunoglobulin (Ig)G-opsonized RBC, or Plasmodium falciparum ring-stage parasitized RBC (RPRBC), degrade ingested hemoglobin rapidly, and can repeat the phagocytic cycle. Monocytes fed with trophozoite-parasitized RBC (TPRBC), which contain malarial pigment, or fed with isolated pigment are virtually unable to degrade the ingested material and to repeat the phagocytic cycle. Monocytes fed with pigment display a long-lasting oxidative burst that does not occur when they phagocytose diamide-treated RBC or RPRBC. The phorbol myristate acetate-elicited oxidative burst is irreversibly suppressed in monocytes fed with TPRBC or pigment, but not in monocytes fed with diamide-treated or IgG-opsonized RBC. This pattern of inhibition of phagocytosis and oxidative burst suggests that malarial pigment is responsible for the toxic effects. Pigment iron released in the monocyte phagolysosome may be the responsible element. 3% of total pigment iron is labile and easily detached under conditions simulating the internal environment of the phagolysosome, i.e., pH 5.5 and 10 microM H2O2. Iron liberated from pigment could account for the lipid peroxidation and increased production of malondialdehyde observed in monocytes fed with pigment or in RBC ghosts and liposomes incubated at pH 6.5 in presence of pigment and low amounts of H2O2. Removal of the labile iron fraction from pigment by repeated treatments with 0.1 mM H2O2 at pH 5.5 reduces pigment toxicity. It is suggested that iron released from ingested pigment is responsible for the intoxication of monocytes. In acute and chronic falciparum infections, circulating and tissue-resident phagocytes are seen filled with TPRBC and pigment particles over long periods of time. Moreover, human monocytes previously fed with TPRBC are unable to neutralize pathogenic bacteria, fungi, and tumor cells, and macrophage responses decline during the course of human and animal malaria. The present results may offer a mechanistic explanation for depression of cellular immunity in malaria.

Adenosine Triphosphate↗

The cone visual pigments of an Australian marsupial, the tammar wallaby (Macropus eugenii): sequence, spectral tuning, and evolution.

Studies on marsupial color vision have been limited to very few species. There is evidence from behavioral, electroretinographic (ERG), and microspectrophotometric (MSP) measurements for the existence of both dichromatic and trichromatic color vision. No studies have yet investigated the molecular mechanisms of spectral tuning in the visual pigments of marsupials. Our study is the first to determine the mRNA sequence, infer the amino acid sequence, and determine, by in vitro expression, the spectra of the cone opsins of a marsupial, the tammar wallaby (Macropus eugenii). This yielded some information on mechanisms and evolution of spectral tuning of these pigments. The tammar wallaby retina contains only short-wavelength sensitive (SWS) and middle-wavelength sensitive (MWS) pigment mRNAs. This predicts dichromatic color vision, which is consistent with conclusions from previous behavioral studies ( Hemmi 1999). We found that the wallaby has a SWS1 class pigment of 346 amino acids. Sequence comparison with eutherian SWS pigments predicts that this SWS1 pigment absorbs maximally (lambdamax) at 424 nm and, therefore, is a blue rather than a UV pigment. This (lambdamax) is close to that of the in vitro-expressed wallaby SWS pigment (lambdamax of 420 +/- 2 nm) and to that determined behaviorally (420 nm). The difference from the mouse UV pigment (lambdamax of 359 nm) is largely accounted for by the F86Y substitution, in agreement with in vitro results comparing a variety of other SWS pigments. This suggests that spectral tuning employing F86Y substitution most likely arose independently in the marsupials and ungulates as a result of convergent evolution. An apparently different mechanism of spectral tuning of the SWS1 pigments, involving five amino acid positions, evolved in primates. The wallaby MWS pigment has 363 amino acids. Species comparisons at positions critical to spectral tuning predict a lambdamax near 530 nm, which is close to that of the in vitro-expressed pigment (529 +/- 1 nm), but quite different from the value of 539 nm determined by microspectrophotometry. Introns interrupt the coding sequences of the wallaby, mouse, and human MWS pigment sequences at the same corresponding nucleotide positions. However, the length of introns varies widely among these species.

Amino Acid Sequence↗

Nonphotosynthetic pigmented bacteria in a potable water treatment and distribution system.

The occurrence of pigmented bacteria in potable water, from raw source water through treatment to distribution water, including dead-end locations, was compared at sample sites in a large municipal water system. Media used to enumerate heterotrophic bacteria and differentiate pigmented colonies were standard method plate count (SPC), m-SPC, and R2A agars, incubated up to 7 days at 35 degrees C. The predominant pigmented bacteria at most sample locations were yellow and orange, with a small incidence of pink organisms at the flowing distribution site. Seasonal variations were seen, with the yellow and orange organisms shifting in dominance. SPC agar was the least productive medium for both heterotroph counts and pigmented bacteria differentiation. At the flowing distribution site, percentages of pigmented bacteria on SPC medium ranged from 2.3 to 9.67 times less than on m-SPC and from 2.3 to 9.86 times less than on R2A. At the same site, seasonal trends in the percentage of pigmented bacteria were the same for m-SPC and R2A media, and the highest and lowest percentages occurred in the fall and winter, respectively. At site 6, there appeared to be an inverse relationship between the yellow and orange pigmented groups, but upon analysis, this did not hold and all correlations between yellow and orange pigmented bacteria were positive. The study results indicate that pigmented bacteria could readily be detected by using plate counting media developed for heterotroph enumeration in potable waters with incubation periods of 7 days. Pigmented bacteria can be used as an additional marker for monitoring changes in water quality. High numbers of heterotrophs, including pigmented forms, were found at dead-end locations, usually in the absence of a free chlorine residual and when the water temperature was greater than 16 degrees C. The association of some pigmented bacteria with nosocomial and other infections raises concern that the organisms may have originated from the potable water supply. High levels of pigmented bacteria could pose an increased health risk to immunologically compromised individuals. Therefore, the bacterial quality of the distribution water should be controlled to prevent the development of high concentrations of heterotrophic plate count bacteria, including the pigmented forms.

Analysis of Variance↗

Direct evidence for the role of pigment cells in the brain of ascidian larvae by laser ablation.

The anterior sensory vesicle of ascidian larvae contains a single large vesicle in which lie two distinct types of pigment cells, anterior and posterior. The ultrastructure of these pigment cells suggests that the anterior pigment cell is an otolith, presumably used for gravity detection, and the posterior pigment cell is an ocellus, used for photoreception. However, there is no direct experimental evidence for this assignment of function. Upward swimming behaviour occurring during the initial period of larval life was examined before and after laser ablation of the anterior pigment and posterior pigment cells. Posterior pigment cell-ablated larvae retained the upward swimming behaviour, but anterior pigment cell-ablated larvae lost it. These results suggest that the anterior pigment acts as a gravity sensor. The negatively phototactic swimming during the latter part of larval life was also examined before and after laser ablation of the anterior pigment or posterior pigment cells. Anterior pigment cell-ablated larvae retained the phototactic response, but posterior pigment cell-ablated larvae lost it. These results suggest that the posterior pigment of the sensory vesicle is involved in the negatively phototactic, downward swimming behavior. The effect of pressure on swimming behaviour was studied, and a putative pressure-detection organ was found not to be involved in the larval swimming behaviour. These are the first published experimental results that permit a functional role in ascidian larval behavior to be assigned to the sensory organs.

Animals↗

Biological and remote sensing perspectives of pigmentation in coral reef organisms.

Coral reef communities face unprecedented pressures on local, regional and global scales as a consequence of climate change and anthropogenic disturbance. Optical remote sensing, from satellites or aircraft, is possibly the only means of measuring the effects of such stresses at appropriately large spatial scales (many thousands of square kilometres). To map key variables such as coral community structure, percentages of living coral or percentages of dead coral, a remote sensing instrument must be able to distinguish the reflectance spectra (i.e. "spectral signature", reflected light as a function of wavelength) of each category. For biotic classes, reflectance is a complex function of pigmentation, structure and morphology. Studies of coral "colour" fall into two disparate but potentially complementary types. Firstly, biological studies tend to investigate the structure and significance of pigmentation in reef organisms. These studies often lack details that would be useful from a remote sensing perspective such as intraspecific variation in pigment concentration or the contribution of fluorescence to reflectance. Secondly, remote sensing studies take empirical measurements of spectra and seek wavelengths that discriminate benthic categories. Benthic categories used in remote sensing sometimes consist of species groupings that are biologically or spectrally inappropriate (e.g. merging of algal phyla with distinct pigments). Here, we attempt to bridge the gap between biological and remote sensing perspectives of pigmentation in reef taxa. The aim is to assess the extent to which spectral discrimination can be given a biological foundation, to reduce the ad hoc nature of discriminatory criteria, and to understand the fundamental (biological) limitations in the spectral separability of biotic classes. Sources of pigmentation in reef biota are reviewed together with remote sensing studies where spectral discrimination has been effectively demonstrated between benthic categories. The basis of reflectance is considered as the sum of pigmented components, such as zooxanthellae, host tissues and skeletons of corals. Problems in the empirical in situ measurement of reflectance are identified, such as the differing types of reflectance which can be measured, the interaction of the light field with morphology, and depth-dependent variability of measured reflectance due to fluorescence. The latter is estimated in some cases to introduce an error of up to 20% when depth differs by 8 m. Spectral features useful in discriminating reef benthos are identified and related to pigmentation. The slope in the reflectance spectra between 650 and 690 nm is dependent on chlorophyll-a concentration and can be used to discriminate bare sand with no algal component from chlorophyll-a containing benthos (algae, corals). The slope in reflectance at various locations between 500 and 560 nm can be useful in discriminating bleached and unbleached corals, possibly due to reduced peridinin concentration. Rhodophyta may be discernible by the presence of a dip in reflectance at 570 nm, due to a phycoerythrin absorption peak. However, the utility of some discriminatory criteria in deeper waters is mitigated by the relatively poor transmission of light through water at longer wavelengths (especially > 600 nm). Contrary to suggested categorizations of fluorescent pigments in coral host tissues, it is shown that these pigments form an almost continuous distribution with respect to their excitation and emission peaks. Remote sensing by induced fluorescence is a promising approach, but further details about the variation and distribution of these pigments are required. It is hoped that this review will promote cross-disciplinary collaboration between pigment biologists and the reef remote sensing community. Where possible, the discriminative criteria adopted in remote sensing should be related to biological phenomena, thus lending an intuitive, process-orientated basis for interpreting spectral data. Similarly, remote sensing may provide a novel scaling perspective to biological studies of pigmentation in reef organisms.

Animals↗

GREEN FLUORESCENT PIGMENT ACCUMULATED BY A MUTANT OF CELLVIBRIO GILVUS.

Love, Samuel H. (Bowman Gray School of Medicine, Winston-Salem, N.C.), and Frank H. Hulcher. Green fluorescent pigment accumulated by a mutant of Cellvibrio gilvus. J. Bacteriol. 87:39-45. 1964.-A mutant of Cellvibrio gilvus, designated strain 139A, liberated a green, fluorescent pigment into the surrounding culture medium. A study of the factors which affected the accumulation of this pigment led to the development of a chemically defined medium which supported maximal pigment accumulation in aerated, liquid cultures. d-Glucose, glycine or l-serine, l-phenylalanine, l-proline, and l-lysine comprised the organic components of this medium. The visible absorption spectrum of the pigment showed a maximal band at 400 mmu (pH 7.0). A difference spectrum between reduced and oxidized pigment showed loss of the band at 400 mmu upon oxidation. However, a methanol-extractable, flavinelike compound occurred in the wild strain but not in the mutant. Ferric ions added to the defined medium stimulated growth, with a concomitant reduction of pigment accumulation. Pigment was formed at a maximal rate during the stationary growth phase, and the highest yield was obtained by 18 hr. Organic solvents did not extract the pigment from water solutions. One and sometimes two, compounds absorbing at 400 mmu could be eluted by ion-exchange chromatography on Cellex-P (H(+)), which was used to separate the pigment from other components in the culture supernatants so that the radioactivity of the pigment could be measured. The mutant synthesized C(14)-labeled pigment from d-glucose-U-C(14) and from each of four amino acids (glycine-1-C(14), l-phenylalanine-U-C(14), l-proline-U-C(14), and l-lysine-U-C(14). Delta-Amino-levulenic acid-4-C(14) did not contribute C(14) to the pigment.

Amino Acids↗

Minocycline-induced pigmentation. Incidence, prevention and management.

Pigmentation is a well recognised adverse effect of minocycline therapy. Various body sites, most notably the skin, nails, bones, thyroid, mouth and eyes are affected and the pigmentation may appear at multiple sites. In general, pigmentation results from long term administration of minocycline at cumulative doses greater than 100 g, although cutaneous or oral mucosal pigmentation may appear, regardless of dose or duration of therapy. When the skin is involved, the blue-black pigmentation develops most frequently on the shins, ankles and arms. Other patterns of skin involvement include pigmentation that is either generalised and symmetrical, or that develops at sites of inflammation. The bones of the oral cavity are probably the most frequently affected sites of pigmentation affecting greater than 20% of patients taking minocycline for more than 4 years. In contrast, the oral mucous membranes and teeth are infrequently pigmented from minocycline. Ocular, thyroid and visceral pigmentation is also relatively uncommon and usually develops only with high doses and long term minocycline use. Whereas pigmentation of the skin and oral mucosa is generally reversible when the drug is discontinued, the pigmentation is often permanent when other sites are involved. Although minocycline-induced pigmentation is not harmful, the drug should be discontinued when the adverse effect is recognised. All patients receiving minocycline, especially those treated for longer than 1 year, require screening for the development of pigmentation.

Anti-Bacterial Agents↗

Ocular pigmentation in white and Siamese cats.

Ocular pigmentation in white cats with blue and yellow eyes and in Siamese cats was examined ophthalmoscopically and histologically. Yellow-eyed white cats had entirely normal ocular pigmentation. Blue eyes of white cats had normal pigmentation of the iridial and retinal pigment epithelia but no stromal pigmentation of the iris or choroid. This deficit is apparently due to the absence of stromal pigment cells, certainly in the iris. As a general rule, the blue eye of white cats had no tapetum. Siamese cats had reduced pigmentation of the iridial and retinal pigment epithelia and no stromal pigmentation of the iris or choroid. The lack of pigmentation is apparently due to the inability of stromal pigment cells to produce pigment, certainly in the iris. We conclude that the abnormality of visual pathways previously described in the Siamese cat is not due simply to a deficiency of pigment of cells of neural crest origin.

Animals↗

On the chemistry of 'black' pigment stones from the gallbladder.

Radiolucent (33 cases) and radiopaque (17 cases) black pigment gallstones from patients who underwent cholecystectomy were studied using several spectroscopic and chromatographic methods. Radiolucent pigment stones (mean Ca percentage 2.1%) are composed chiefly of degenerated tetrapyrrolic bile pigments (mean 85.3%) deriving from bilirubin and bilirubinates. Degeneration includes both polymerization and bacterial reduction and leads to products of different grade of polymerization. Final extraction residues (mean 55.5%), called the 'black pigments' are considered to be degenerated bile pigments of high molecular weight. The mean percentage of bilirubin (free and inorganic bound bilirubin) was 8.5%, while the percentage of lipids was very low (mean of total lipids approximately 2.7%). Radiopaque black pigment stones (Ca: 12.4%) were composed of 'black pigments', too, but contained large amounts of calcium phosphate (carbonate apatite) and/or calcium carbonate. 65% of the radiopaque stones were calcified by calcium phosphate. 'Black pigments' were degraded by chromate to maleimides and 2,5-pyrroledialdehyde. These degradation products can be prepared in the same way from normal bile pigments with a tetrapyrrole structure. Polymerized dipyrrolic bile pigments like polymer propentdyopent or 'mesobilifuscin' did not give 2,5-pyrroledialdehydes during chromate oxidation. Thus we conclude that the formation of 'black pigments' starts from the polymerization of tetrapyrrolic, but not from dipyrrolic units. Accumulation of unconjugated bilirubin and bilirubinates within the gallbladder will precede the development of 'black pigments' which play an important role in pigment gallstone formation.

Bile↗

Destruction of liver haem by norethindrone. Conversion into green pigments.

1. Factors affecting the norethindrone-mediated conversion of hepatic haem into green pigments have been studied in the rat. Concentrations of haem and green pigments were estimated spectrophotometrically after esterification and separation by silica gel high-pressure liquid chromatography (h.p.l.c.). 2. Accumulation of green pigments in the liver was dependent on the dose of steroid and the time after dosing, maximum values being reached after 4-8h. Phenobarbitone pretreatment of rats resulted in an 8-fold increase in the concentration of green pigments at these times. 3. In microsomal systems in vitro, the formation of green pigments in the presence of NADPH and norethindrone was also dependent on the concentration of steroid and incubation times. Reaction rates very rapidly became non-linear with time, consistent with the self-catalysed destruction of the form(s) of cytochrome P-450 responsible for the metabolic activation of norethindrone. Microsomal mixtures incubated for a short period of time (1min) with norethindrone gave only one green-pigment peak after h.p.l.c. Longer incubation times gave four or five additional green pigments. Results suggested that multiple green pigments may arise by metabolic transformation of a single precursor. 4. When liver haem was prelabelled with (14)C by using 5-amino[4-(14)C]laevulinic acid, subsequent dosing with norethindrone in vivo gave rise to three major (14)C-labelled-green-pigment peaks on h.p.l.c. None of these components had the same retention times as the green pigments produced by microsomal fractions in vitro. 5. When liver haem was prelabelled with (59)Fe by using (59)FeCl(3), norethindrone administration resulted in the detection of (59)Fe-labelled green pigments if subsequent esterification was carried out under neutral conditions with trimethyloxonium tetrafluoroborate, but not when carried out under acidic conditions with methanol/H(2)SO(4). These results suggested that green pigments normally contain chelated iron and that metal-free green pigments are not produced by the liver.

Animals↗

Clinical signs of the pigment dispersion syndrome in blacks.

BACKGROUND: The pigment dispersion syndrome (PDS) is considered rare in blacks, and minimal literature exists concerning the condition in this patient population. The diagnosis of PDS in blacks may present unique challenges because some of the typical clinical signs that are present in whites, including iris transillumination defects, posterior iris bowing, and noticeable anterior iris stromal pigment dusting, may not occur as commonly. Diagnosis can be particularly difficult when neither these signs nor significant corneal endothelial pigmentation exists. Although zonular and peripheral lens pigment has been found to be consistently present in whites with PDS, attention has not been given to this as a potentially important diagnostic sign in blacks. METHODS: From among a primary care population, we identified and studied 7 patients (13 eyes) who exhibited moderate to heavy trabecular meshwork (TM) pigmentation, as well as zonule and/or peripheral lens pigmentation. Patients were identified during routine clinical care provided by one of the authors, as well as from notification by other practitioners. All patients received complete eye examination and other signs of PDS were looked for. RESULTS: Four males and 3 females were identified, their average age being 37 years (range = 15 to 51) at the time of their initial identification. All but one patient was myopic (average approximately -2.50 D spherical equivalent). Iris transillumination defects were present in only one eye of one patient, and no eyes showed overt posterior iris bowing, although the iris contours were usually flat and the anterior chambers appeared relatively deep. Corneal endothelial pigmentation was frequently barely detectable and could not be relied on as a predictor of trabecular meshwork or lenticular pigmentation. Glaucoma, or a suspicion of glaucoma due to increased intraocular pressure (IOP) or cupping, was common among the group. Using heavy TM pigmentation as well as any degree of zonular and/or peripheral lenticular pigmentation as a criteria for the diagnosis of PDS, we calculated the prevalence of PDS among blacks in a nonreferred primary care population (> age 7) to be at least 15 cases per 10,000. CONCLUSIONS: More investigation is needed to study the clinical presentation of PDS in blacks because it may be substantially different than in whites. Zonular and peripheral lenticular pigmentation may be a particularly useful diagnostic sign of PDS in blacks, especially in those cases where other traditional signs, including iris transillumination defects, pronounced corneal endothelial pigmentation, posterior iris bowing, and visible anterior iris stromal pigment dusting, are absent. The "classic" variety of PDS may be more common among blacks than previously recognized.

Adolescent↗