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Sample records for cultured pure melanocytes

  1. Isolation, culture, and transfection of melanocytes.

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    Godwin, Lauren S; Castle, Joanna T; Kohli, Jaskaren S; Goff, Philip S; Cairney, Claire J; Keith, W Nicol; Sviderskaya, Elena V; Bennett, Dorothy C

    2014-06-03

    Located in the basal epidermis and hair follicles, melanocytes of the integument are responsible for its coloration through production of melanin pigments. Melanin is produced in lysosomal-like organelles called melanosomes. In humans, this skin pigmentation acts as an ultraviolet radiation filter. Abnormalities in the division of melanocytes are quite common, with potentially oncogenic growth usually followed by cell senescence producing benign naevi (moles), or occasionally melanoma. Therefore, melanocytes are a useful model for studying melanoma, as well as pigmentation and organelle transport and the diseases affecting these mechanisms. This chapter focuses on the isolation, culture, and transfection of human and murine melanocytes. The first basic protocol describes the primary culture of melanocytes from human skin and the maintenance of growing cultures. The second basic protocol details the subculture and preparation of mouse keratinocyte feeder cells. The primary culture of melanocytes from mouse skin is described in the third basic protocol, and, lastly, the fourth basic protocol outlines a technique for transfecting melanocytes and melanoma cells.

  2. Detection of auto antibodies and transplantation of cultured autologous melanocytes for the treatment of vitiligo

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    Zhu, Mei-Cai; Ma, Hong-Yu; Zhan, Zhi; Liu, Cheng-Gang; Luo, Wei; Zhao, Guang

    2017-01-01

    The aim of the present study was to establish an immunofluorescence method of antibody detection to identify melanocytes in the serum of vitiligo patients. Furthermore, we aimed to establish a method for the culture and proliferation of autologous pure melanocytes and to observe the effect of their transplantation for the treatment of vitiligo. Suspension of epidermal cells with melanocytes was performed using trypsin digestion of normal epiderm from eyelid operation and melanocytes were selectively cultured and proliferated in serum-free M2 medium. FITC-labeled rabbit anti-human antibody was used to detect the relative fluorescence intensity of the melanocytes. After identification with immunological and biological examinations, the melanocytes were transplanted to depigmented areas of vitiligo. Repigmentation was observed continuously. The results indicated that melanocytes could be selectively proliferated in the medium. Subsequently, pure melanocytes without contamination of fibroblast and keratinocyte were harvested. A total of 34 patients suffering vitiligo for between 3 months and 20 years with depigmented area (between 4 cm2 and 70% of body surface) were divided into 19 cases of developing stage and 15 cases of stable stage, according to the change of depigmentation. A total of 15 developing cases were positive for the antibody against melanocytes, with the positive rate of 79%. The titers of serum was >1:50 in 10 patients at the developing stage, and 5 developing patients were 1:10. Among the 15 stable cases, four were positive, with a positive rate of 27%. Fluorescence of antibody was localized in the cytoplasm of the melanocytes. Autologous melanocytes of vitiligo patients could be selectively proliferated in the medium. Next, pure melanocytes without contamination with fibroblasts and keratinocytes were harvested. A total of 16 vitiligo patients with 28 depigmented areas (2–200 cm2) were treated with transplantation of melanocytes. Repigmentation of

  3. Effects of hydroxybenzyl alcohols on melanogenesis in melanocyte-keratinocyte co-culture and monolayer culture of melanocytes.

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    Liu, Szu-Hsiu; Chu, I-Ming; Pan, I-Horng

    2008-08-01

    In mammalian skin, melanocyte proliferation and melanogenesis can be stimulated by keratinocytes, fibroblasts and other regulatory factors. To determine whether hydroxybenzyl alcohols (HBAs) show more inhibitory in melanocytes cultured alone or in melanocytes co-cultured with keratinocytes, we developed a murine melanocyte-keratinocyte co-culture model to investigate the pigmentation regulators in company with other melanogenic inhibitors and stimulators. It was found that the effects of HBAs and melanogenic factors were more evident in melanocytes co-cultured with keratinocytes. Keratinocytes may play a synergistic role in melanocyte melanogenesis and influence the pigment production. The tests in the co-culture model also imply that the inhibitory effects of HBAs on melanogenesis are due to the direct inhibition of melanosomal tyrosinase activity. HBAs showed a low cytotoxicity. The eventual results proved that HBAs are promising and safe agents for skin whitening in melanocyte alone and in co-culture systems. The co-culture model provides a more physiologically realistic condition to study the interaction between melanocytes and keratinocytes, which enables a reliable screening system for depigmenting compounds.

  4. Hair Follicle Melanocyte Cells as a Renewable Source of Melanocytes for Culture and Transplantation

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    Kwon, Ho

    2008-01-01

    Full Text Available Objective: Advances in melanocyte culture techniques have not yet led to reliable clinical methods for treating hypopigmentation disorders. We hypothesized that melanocytes harvested from plucked hair follicles may provide a renewable source of melanocytes for the treatment of hypopigmentation. Methods: Hairs with attached cells from the follicles were plucked from Yucatan pigs and implanted in a collagen-glycosaminoglycan matrix for either immediate or delayed implantation into full-thickness excisional porcine wounds. Wounds were allowed to heal and were biopsied at 2 and 4 weeks, respectively. Results: Fully healed wounds with transplanted hair follicles showed central areas of dark pigmentation corresponding to the location of implanted hair follicles. Corresponding collagen-glycosaminoglycan matrix wounds showed no central areas of pigmentation. Conclusions: Hair follicle--derived melanocytes may potentially serve as a renewable source of pigment-producing cells for treating hypopigmentation disorders.

  5. Effects of diethylstilbestrol on the proliferation and tyrosinase activity of cultured human melanocytes

    OpenAIRE

    TANG, JIANBING; Li, Qin; Cheng, Biao; HUANG, CHONG; Chen, Kui

    2015-01-01

    The aim of the present study was to observe the effects of different exogenous estrogen diethylstilbestrol (DES) concentrations on the human melanocyte proliferation and tyrosinase activity. Skin specimens were obtained following blepharoplasty, and the melanocytes were primary cultured and passaged to the third generation. The melanocytes were seeded in 96-well plates, each well had 5×103 cells. The medium was changed after 24 h, and contained 10−4-10−8 M DES. After the melanocytes were incu...

  6. Primary culture of human skin melanocyte and comparison of culture in the presence and absence of phorbol ester

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    Reza Yarani

    2013-06-01

    Full Text Available Background: Primary culture takes place following the cell isolation from tissues. Isolation and culture of melanocytes based on their roll in the protection of body against hazardous sun rays, production of skin, cornea and hair color is really important. This study was done to set isolation, culture and proliferation of melanocytes from children foreskin and adult eyelashes, and also comparison of two types of melanocyte culture medium. Methods: Human foreskin and eyelash samples were used for melanocyte isolation and culture. After isolation of epidermis from dermis, epidermis cell suspensions were prepared by enzymatic digestion. The isolated cells were cultured in two melanocyte selective culture media. Immunocytochemistary and reverse transcription-polymerase chain reaction (RT-PCR assays were used for confirmation of isolated and cultured melanocytes. Results: Our results indicated that isolated melanocyte cultured in the selective medium without phorbol esters is better than the melanocytes cultured in selective medium cont-aining phorbol esters not only morphologically but also physiologically and from the aspect of cell adhesion. In addition, the results showed that isolated melanocyte from adult eyelashes are more dendritic than melanocytes isolated from children foreskin. Conversely, our results indicated that the number of cell passages in melanocyte isolat-ed from foreskin is more than melanocytes isolated from adult eyelashes. Conclusion: Melanocytes cultured in selective medium containing convenient growth factors in absence of phorbol esters show more native physiological and adhesive properties. In addition, melanocyte isolated from younger tissues such as foreskin have better proliferative and sub-culturing properties so we suggest isolation and culture of younger tissues.

  7. Endothelin-1 increases melanin synthesis in an established sheep skin melanocyte culture.

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    Pang, Yamiao; Geng, Jianjun; Qin, Yilong; Wang, Haidong; Fan, Ruiwen; Zhang, Ying; Li, Hongquan; Jiang, Shan; Dong, Changsheng

    2016-08-01

    The aims of the study were to establish a culture system for sheep skin melanocytes and uncover the effects of endothelin-1 on melanin synthesis in cultured melanocytes in order to provide an optimal cell system and a theoretical basis for studying the regulatory mechanism of coat color in sheep. In this study, skin punch biopsies were harvested from the dorsal region of 1-3-yr-old sheep, and skin melanocytes were then obtained by the two-step digestion using dispase II and trypsin/ethylene diamine tetraacetic acid (EDTA). The primary cultures of the melanocytes were established and characterized by dopa-staining, immunocytochemical localization of melanocyte markers, and RT-polymerase chain reaction (PCR) analysis of coat color genes. To determine the effect of endothelin-1 on proliferation and melanin synthesis of melanocytes, the cultured cells were treated with different doses of endothelin-1 (10(-7), 10(-8), 10(-9), 10(-10), and 0 mol/L), and the growth rate of melanocytes, production of melanin, expression of related genes, and location of related protein in cultured cells were examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), ultraviolet spectrophotometry, qRT-PCR, and immunocytochemical localization, respectively. The results showed that the established melanocyte culture functions properly. Endothelin-1 treatment increased markedly the number of melanocytes and melanin content. In responding to this treatment, expressions of microphthalmia-associated transcription factor (MITF), melanocortin 1 receptor (MC1R), tyrosinase (TYR), and endothelin receptor B (EDNRB) in the melanocytes were significantly up regulated (P skin melanocytes was established successfully in vitro, and endothelin-1 promotes the melanogenesis in sheep skin melanocytes.

  8. THE EFFECT OF Ge-132 ON ULTRASTRUCTURE OF CULTURED MELANOCYTES

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Objective To elucidate the effect of Ge-132 on the growth of melanocytes. Mothods Melanocyes from epidermis were cultured and purified ;the second generation of the cell was used for study ;the cells were divided into two groups randomly,to group A, Ge-132 was added to the media at 0.04mg/L ;to group B ,common culturing method was used without Ge-132. After 5d, the cells were seperated by digestion for study by transmission electronic micro- scope. Results Compared to group B, the vacuioes of the cells were increased,mitochondria distended, endoplasmic reticulum dilated and the number of melanosome declined in the group A. Conclusion Ge-132 can inhibit the melanocyte's growth at a certain concentration and might be used for treating pigmented diseases.

  9. Bringing Planctomycetes into pure culture

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    Olga Maria Lage

    2012-12-01

    Full Text Available Planctomycetes have been known since the description of Planctomyces bekefii by Gimesi at the beginning of the twentieth century (1924, although the first axenic cultures were only obtained in the 1970s. Since then, eleven genera with fourteen species have been validly named and five candidatus genera belonging to the anaerobic ammonium oxidation, anammox bacteria have also been discovered. However, Planctomycetes diversity is much broader than these numbers indicate, as shown by environmental molecular studies. In recent years the authors have attempted to isolate and cultivate additional strains of Planctomycetes. This paper provides a summary of the isolation work that was carried out to obtain in pure culture Planctomycetes from several environmental sources. The following strains and planctomycetes have been successfully isolated: two freshwater strains from the sediments of an aquarium, which were described as a new genus and species, Aquisphaera giovannonii; several Rhodopirellula strains from the sediments of a water treatment recycling tank of a marine fish farm; and more than 140 planctomycetes from the biofilm community of macroalgae. This collection comprises several novel taxa that are being characterized and described. Improvements in the isolation methodology were made in order to optimize and enlarge the number of Planctomycetes isolated from the macroalgae. The existence of an intimate and an important relationship between planctomycetes and macroalgae reported before by molecular studies is therefore supported by culture dependent methods.

  10. Post-transcriptional regulation of neurofibromin level in cultured human melanocytes in response to growth factors.

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    Griesser, J; Kaufmann, D; Maier, B; Mailhammer, R; Kuehl, P; Krone, W

    1997-03-01

    Among the symptoms that characterize neurofibromatosis type 1 (NF1) are pigmentation anomalies such as cafe au lait spots. It has been suggested that the reduction of the neurofibromin level in the epidermis of NF1 patients is responsible for the observed signs such as altered melanogenesis and altered density of melanocytes. Our studies show that in cultured normal human melanocytes, the neurofibromin level can be varied in vitro over a wide range by using different culture conditions. The influence of factors that control differentiation and proliferation of melanocytes on neurofibromin levels was studied. Immunoprecipitation followed by western blotting showed a 3- to 4-fold increase of neurofibromin after stimulation by PMA or bFGF, respectively, and a 1.5-fold increase in cells stimulated with steel factor. The increase of neurofibromin was not paralleled by a higher NF1 mRNA level as proved by northern blotting. Pulse-chase experiments with 35S-labeled melanocytes revealed an approximately 3-fold increase in the half-life of neurofibromin in bFGF- or PMA-stimulated cells compared to controls. These results indicate that the neurofibromin level of cultured melanocytes can be regulated by a mechanism independent of NF1 gene transcription and translation, which might influence the degradation rate of the protein.

  11. Dark fermentation on biohydrogen production: Pure culture.

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    Lee, Duu-Jong; Show, Kuan-Yeow; Su, Ay

    2011-09-01

    Biohydrogen is regarded as an attractive future clean energy carrier due to its high energy content and environmental-friendly conversion. While biohydrogen production is still in the early stage of development, there have been a variety of laboratory- and pilot-scale systems developed with promising potential. This work presents a review of literature reports on the pure hydrogen-producers under anaerobic environment. Challenges and perspective of biohydrogen production with pure cultures are also outlined.

  12. (Pheo)melanin photosensitizes UVA-induced DNA damage in cultured human melanocytes

    NARCIS (Netherlands)

    Wenczl, E.; Schans, G.P. van der; Roza, L.; Kolb, R.M.; Timmerman, A.J.; Smit, N.P.M.; Pavel, S.; Schothorst, A.A.

    1998-01-01

    The question of whether melanins are photoprotecting and/or photosensitizing in human skin cells continues to be debated. To evaluate the role of melanin upon UVA irradiation, DNA single-strand breaks (ssb) were measured in human melanocytes differing only in the amount of pigment produced by cultur

  13. Biological characteristics of mouse skin melanocytes.

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    Shi, Zhanquan; Ji, Kaiyuan; Yang, Shanshan; Zhang, Junzhen; Yao, Jianbo; Dong, Changsheng; Fan, Ruiwen

    2016-04-01

    The objective of this research was to evaluate the optimal passage number according to the biological characteristics of mouse skin melanocytes from different passages. Skin punch biopsies harvested from the dorsal region of 2-day old mice were used to establish melanocyte cultures. The cells from passage 4, 7, 10 and 13 were collected and evaluated for their melanogenic activity. Histochemical staining for tyrosinase (TYR) activity and immunostaining for the melanocyte specific markers including S-100 antigen, TYR, tyrosinase related protein 1 (TYRP1), tyrosinase related protein 2 (TYRP2) and micropthalmia associated transcription factor (MITF) confirmed purity and melanogenic capacity of melanocytes from different passages, with better melanogenic activity of passage 10 and 13 cells being observed. Treatment of passage 13 melanocytes with α-melanocyte stimulating hormone (α-MSH) showed increased expression of MITF, TYR and TYRP2 mRNA. However, considering the TYR mRNA dramatically high expression which is the characteristics of melanoma cells, melanocytes from passage 10 was the optimal passage number for the further research. Our results demonstrate culture of pure populations of mouse melanocytes to at least 10 passages and illustrate the potential utility of passage 10 cells for studies of intrinsic and extrinsic regulation of genes controlling pigmentation and coat color in mouse.

  14. Development of melanocye-keratinocyte co-culture model for controls and vitiligo to assess regulators of pigmentation and melanocytes

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    Ravinder Kumar

    2012-01-01

    Full Text Available Background: There is a need to develop an in vitro skin models which can be used as alternative system for research and testing pharmacological products in place of laboratory animals. Therefore to study the biology and pathophysiology of pigmentation and vitiligo, reliable in vitro skin pigmentation models are required. Aim: In this study, we used primary cultured melanocytes and keratinocytes to prepare the skin co-culture model in control and vitiligo patients. Methods: The skin grafts were taken from control and patients of vitiligo. In vitro co-culture was prepared after culturing primary melanocytes and keratinocytes. Co- cultures were treated with melanogenic stimulators and inhibitors and after that tyrosinase assay, MTT assay and melanin content assay were performed. Results: Melanocytes and keratinocytes were successfully cultured from control and vitiligo patients and after that co-culture models were prepared. After treatment of co-culture model with melanogenic stimulator we found that tyrosinase activity, cell proliferation and melanin content increased whereas after treatment with melanogenic inhibitor, tyrosinase activity, cell proliferation and melanin content decreased. We also found some differences in the control co-culture model and vitiligo co-culture model. Conclusion: We successfully constructed in vitro co-culture pigmentation model for control and vitiligo patients using primary cultured melanocytes and keratinocytes. The use of primary melanocytes and keratinocytes is more appropriate over the use of transformed cells. The only limitation of these models is that these can be used for screening small numbers of compounds.

  15. In vitro dedifferentiation of melanocytes from adult epidermis.

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    Bernadett Kormos

    Full Text Available In previous work we described a novel culture technique using a cholera toxin and PMA-free medium (Mel-mix for obtaining pure melanocyte cultures from human adult epidermis. In Mel-mix medium the cultured melanocytes are bipolar, unpigmented and highly proliferative. Further characterization of the cultured melanocytes revealed the disappearance of c-Kit and TRP-1 and induction of nestin expression, indicating that melanocytes dedifferentiated in this in vitro culture. Cholera toxin and PMA were able to induce c-Kit and TRP-1 protein expressions in the cells, reversing dedifferentiation. TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression. These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation. Because melanocytes in Mel-mix medium can be cultured with human serum as the only supplement, this culture system is also suitable for autologous cell transplantation.

  16. In vitro dedifferentiation of melanocytes from adult epidermis.

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    Kormos, Bernadett; Belso, Nóra; Bebes, Attila; Szabad, Gábor; Bacsa, Sarolta; Széll, Márta; Kemény, Lajos; Bata-Csörgo, Zsuzsanna

    2011-02-23

    In previous work we described a novel culture technique using a cholera toxin and PMA-free medium (Mel-mix) for obtaining pure melanocyte cultures from human adult epidermis. In Mel-mix medium the cultured melanocytes are bipolar, unpigmented and highly proliferative. Further characterization of the cultured melanocytes revealed the disappearance of c-Kit and TRP-1 and induction of nestin expression, indicating that melanocytes dedifferentiated in this in vitro culture. Cholera toxin and PMA were able to induce c-Kit and TRP-1 protein expressions in the cells, reversing dedifferentiation. TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression. These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation. Because melanocytes in Mel-mix medium can be cultured with human serum as the only supplement, this culture system is also suitable for autologous cell transplantation.

  17. Pure Culture Fermentation of Brined Cucumbers.

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    Etchells, J L; Costilow, R N; Anderson, T E; Bell, T A

    1964-11-01

    The relative abilities of Pediococcus cerevisiae, Lactobacillus plantarum, L. brevis, and several other species of lactic acid bacteria to grow and produce acid in brined cucumbers were evaluated in pure culture fermentations. Such fermentations were made possibly by the use of two techniques, gamma radiation (0.83 to 1.00 Mrad) and hot-water blanching (66 to 80 C for 5 min), designed first to rid the cucumbers of naturally occurring, interfering, and competitive microbial groups prior to brining, followed by inoculation with the desired lactic acid bacteria. Of the nine species tested, strains of the three common to cucumber fermentations, P. cerevisiae, L. plantarum, and L. brevis, grew to the highest populations, and produced the highest levels of brine acidity and the lowest pH values in fermentations at 5.4 to 5.6% NaCl by weight; also, their sequence of active development in fermentations, with the use of a three-species mixture for inoculation, was in the species order just named. This sequence of occurrence was similar to that estimated by others for natural fermentations. The rates of growth and acid production in fermentations with a mixture of P. cerevisiae, L. plantarum, and L. brevis increased as the incubation temperature was increased from 21 to 27 to 32 C; however, the maximal populations and acidities attained were essentially the same for fermentations at each temperature. Further, these same three species were found to be the most salt tolerant of those tested; their upper limit for appreciable growth and measurable acid production was about 8% salt, whereas thermophilic species such as L. thermophilus, L. lactis, L. helveticus, L. fermenti, and L. delbrueckii exhibited a much lower salt tolerance, ranging from about 2.5 to 4.0%. However, certain strains of L. delbrueckii grew very rapidly in cucumbers brined at 2.5 to 3.0% salt, and produced sufficient acid in about 30 hr at 48 C to reduce the brine pH from above 7.0 to below 4.0. An inexpensive

  18. Gene expression profiling of cultured human NF1 heterozygous (NF1+/-) melanocytes reveals downregulation of a transcriptional cis-regulatory network mediating activation of the melanocyte-specific dopachrome tautomerase (DCT) gene.

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    Boucneau, Joachim; De Schepper, Sofie; Vuylsteke, Marnik; Van Hummelen, Paul; Naeyaert, Jean-Marie; Lambert, Jo

    2005-08-01

    One of the major primary features of the neurocutaneous genetic disorder Neurofibromatosis type 1 are the hyperpigmentary café-au-lait macules where disregulation of melanocyte biology is supposed to play a key etiopathogenic role. To gain better insight into the possible role of the tumor suppressor gene NF1, a transcriptomic microarray analysis was performed on human NF1 heterozygous (NF1+/-) melanocytes of a Neurofibromatosis type 1 patient and NF1 wild type (NF1+/+) melanocytes of a healthy control patient, both cultured from normally pigmented skin and hyperpigmented lesional café-au-lait skin. From the magnitude of gene effects, we found that gene expression was affected most strongly by genotype and less so by lesional type. A total of 137 genes had a significant twofold or more up- (72) or downregulated (65) expression in NF1+/- melanocytes compared with NF1+/+ melanocytes. Melanocytes cultured from hyperpigmented café-au-lait skin showed 37 upregulated genes whereas only 14 were downregulated compared with normal skin melanocytes. In addition, significant genotype xlesional type interactions were observed for 465 genes. Differentially expressed genes were mainly involved in regulating cell proliferation and cell adhesion. A high number of transcription factor genes, among which a specific subset important in melanocyte lineage development, were downregulated in the cis-regulatory network governing the activation of the melanocyte-specific dopachrome tautomerase (DCT) gene. Although the results presented have been obtained with a restricted number of patients (one NF1 patient and one control) and using cDNA microarrays that may limit their interpretation, the data nevertheless addresses for the first time the effect of a heterozygous NF1 gene on the expression of the human melanocyte transcriptome and has generated several interesting candidate genes helpful in elucidating the etiopathology of café-au-lait macules in NF1 patients.

  19. Pure Culture Fermentation of Green Olives1

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    Etchells, J. L.; Borg, A. F.; Kittel, I. D.; Bell, T. A.; Fleming, H. P.

    1966-01-01

    The method previously developed by us for the pure-culture fermentation of brined cucumbers and other vegetables has been applied successfully to Manzanillo variety olives. Field-run grade fruit was processed first by conventional procedures to remove most of the bitterness. Then the relative abilities of Lactobacillus plantarum, L. brevis, Pediococcus cerevisiae, and Leuconostoc mesenteroides to become established and produce acid in both heat-shocked (74 C for 3 min) and unheated olives, brined at 4.7 to 5.9% NaCl (w/v basis), were evaluated. The heat-shock treatment not only proved effective in ridding the fruit of naturally occurring, interfering, and competitive microbial groups prior to brining and inoculation, but also made the olives highly fermentable with respect to growth and acid production by the introduced culture, particularly L. plantarum. Of the four species used as inocula, L. plantarum was by far the most vigorous in fermentation ability. It consistently produced the highest levels of brine acidity (1.0 to 1.2% calculated as lactic acid) and the lowest pH values (3.8 to 3.9) during the fermentation of heat-shocked olives. Also, L. plantarum completely dominated fermentations when used in two-species (with P. cerevisiae) and three-species (with P. cerevisiae and L. brevis) combinations as inocula. In contrast, when L. plantarum was inoculated into the brines of unheated olives it failed to become properly established; the same was true for the other species tested, but even to a more pronounced degree. L. brevis was the only species used that failed to develop in brines of both heat-shocked and unheated olives. Modification of the curing brine by the addition of lactic acid at the outset, either with or without dextrose, led to a much earlier onset of fermentation with accompanying acid development, as compared to treatments with dextrose alone or nonadditive controls. Reasons for the marked improvement of the fermentability of Manzanillo olives

  20. Melanin offers protection against induction of cyclobutane pyrimidine dimers and 6-4 photoproducts by UVB in cultured human melanocytes

    NARCIS (Netherlands)

    Smit, N.P.M.; Vink, A.A.; Kolb, R.M.; Steenwinkel, M.J.S.T.; Berg, P.T.M. van den; Nieuwpoort, F. van; Roza, L.; Pavel, S.

    2001-01-01

    The goal of this investigation was to correlate the melanin content in human pigmentary cells with the generation of UVB-induced photoproducts and to examine the relationship between the melanin content and the removal of the photoproducts. Cultured melanocytes from light-skinned individuals

  1. Melanin offers protection against induction of cyclobutane pyrimidine dimers and 6-4 photoproducts by UVB in cultured human melanocytes

    NARCIS (Netherlands)

    Smit, N.P.M.; Vink, A.A.; Kolb, R.M.; Steenwinkel, M.J.S.T.; Berg, P.T.M. van den; Nieuwpoort, F. van; Roza, L.; Pavel, S.

    2001-01-01

    The goal of this investigation was to correlate the melanin content in human pigmentary cells with the generation of UVB-induced photoproducts and to examine the relationship between the melanin content and the removal of the photoproducts. Cultured melanocytes from light-skinned individuals synthes

  2. Development of melanocye-keratinocyte co-culture model for controls and vitiligo to assess regulators of pigmentation and melanocytes

    OpenAIRE

    2012-01-01

    Background: There is a need to develop an in vitro skin models which can be used as alternative system for research and testing pharmacological products in place of laboratory animals. Therefore to study the biology and pathophysiology of pigmentation and vitiligo, reliable in vitro skin pigmentation models are required. Aim: In this study, we used primary cultured melanocytes and keratinocytes to prepare the skin co-culture model in control and vitiligo patients. Methods: The skin grafts wer...

  3. Comparison of growth factor signalling pathway utilisation in cultured normal melanocytes and melanoma cell lines

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    Kim Ji Eun

    2012-04-01

    Full Text Available Abstract Background The phosphatidylinositol-3-kinase (PI3K-PKB, mitogen activated protein kinase (MEK-ERK and the mammalian target of rapamycin (mTOR- p70S6K, are thought to regulate many aspects of tumour cell proliferation and survival. We have examined the utilisation of these three signalling pathways in a number of cell lines derived from patients with metastatic malignant melanoma of known PIK3CA, PTEN, NRAS and BRAF mutational status. Methods Western blotting was used to compare the phosphorylation status of components of the PI3K-PKB, MEK-ERK and mTOR-p70S6K signalling pathways, as indices of pathway utilisation. Results Normal melanocytes could not be distinguished from melanoma cells on the basis of pathway utilisation when grown in the presence of serum, but could be distinguished upon serum starvation, where signalling protein phosphorylation was generally abrogated. Surprisingly, the differential utilisation of individual pathways was not consistently associated with the presence of an oncogenic or tumour suppressor mutation of genes in these pathways. Conclusion Utilisation of the PI3K-PKB, MEK-ERK and mTOR-p70S6K signalling pathways in melanoma, as determined by phosphorylation of signalling components, varies widely across a series of cell lines, and does not directly reflect mutation of genes coding these components. The main difference between cultured normal melanocytes and melanoma cells is not the pathway utilisation itself, but rather in the serum dependence of pathway utilisation.

  4. Selective proliferation of normal human melanocytes in vitro in the presence of phorbol ester and cholera toxin by Eisinger and Marko.

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    Scott, Glynis

    2014-01-01

    Melanocytes are pigment producing cells that arise from the neural crest and migrate to the skin early in fetal development. The pigment that melanocytes synthesize, melanin, plays a critical role in protecting the skin from mutagenic ultraviolet irradiation. Melanocytes are also precursor cells for melanoma, a deadly form of skin cancer. Because melanocytes make up a minority population of cells in the epidermis they have been difficult to propagate in culture. The landmark paper by Eisinger and Marko, described below, was the first successful report of large scale propagation of pure cultures of melanocytes. This paper set the stage for an explosive growth in knowledge in the biology of human melanocytes and allowed scientists to begin dissecting the different oncogenic events involved in the transition of melanocytes to melanoma.

  5. Study of cultured bovine capsular bag in pure ocular tissue

    Institute of Scientific and Technical Information of China (English)

    WANG Yan-qing; LI Qiu-ming

    2006-01-01

    @@ The proliferation, differentiation and fibrosis of lens epithelia cells (LECs) is mainly responsible for posterior capsular opacification (PCO). From the primary culture of LECs to the culture of lens capsular bag, the models of posterior capsular opacification have been developed. At present, the most commonly used model is cell culture in medium with serum. But the culture in pure ocular tissue has not been reported. Therefore, we established a new model of posterior capsular opacification-culturing bovine lens capsular bag in pure ocular tissue to exclude the role of serum. Our study established a new culture method to investigate the proliferation,differentiation and apoptosis of lens epithelia cells in the aqueous humor with or without lens cortex and vitreous humor. The purpose of the study is to model posterior capsular opacification in vivo as closely as possible and to discuss the influence of ocular tissue on posterior capsular opacification.

  6. Pure Land or Pure Mind?: Locus of Awakening and American Popular Religious Culture

    Directory of Open Access Journals (Sweden)

    Richard K. Payne

    2015-03-01

    Full Text Available This essay has two sections, each with its own distinct goal, forming an interrelated whole. The first introduces “locus of awakening,” and applies it to the relative success in America of Zen and Tibetan Buddhisms, compared to Pure Land Buddhism. The explanatory power of the concept is demonstrated by also considering Soka Gakkai. The difference between popular culture treatments of Zen and Tibetan Buddhisms, and Pure Land Buddhism was the problematic leading to identifying locus of awakening as an aspect of Buddhist thought. The second section locates it in the history of Buddhist thought, demonstrating that it is not a modern conceptualization of the path, not one created in response to Euro–American religio-therapeutic culture. Locus of awakening is, instead, part of the continuity of the Buddhist tradition, and does not fall on one side or the other of the sometimes overdrawn dichotomy between Asian and American Buddhisms.

  7. Bioleaching of chalcopyrite by pure and mixed culture

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yan-sheng; QIN Wen-qing; WANG Jun; ZHEN Shi-jie; YANG Cong-ren; ZHANG Jian-wen; NAI Shao-shi; QIU Guan-zhou

    2008-01-01

    The bioleaching of chalcopyrite in shake flasks was investigated by using pure Acidithiobacillus ferrooxidans and mixed culture isolated from the acid mine drainage in Yushui and Dabaoshan Copper Mine in China,marked as YS and DB,respectively.The mixed culture consisted mainly of Acidithiobacillus ferrooxidans,Acidithiobacillus thiooxidans,and Leptospirillum spp.(Leptospirillum ferriphilum and Leptospirillum ferrooxians).The results show that the mixed culture is more efficient than the pure Acidithiobacillus ferrooxidans because of the presence of the sulfur-oxidizing cultures that positively increase the dissolution rate and the recovery of copper from chalcopyrite.The pH value decreases with the decrease of chalcopyrite leaching rate,because of the formation of jarosite as a passivation layer on the mineral surface during bioleaching.In the bioleaching using the mixed culture,low pH is got from the sulfur oxidizing inhibiting,the formation of jarosite.The copper extraction reaches 46.27% in mixed culture and 30.37% in pure Acidithiobacillus ferrooxidans after leaching for 75 d.

  8. 人黑素细胞体外培养条件的研究进展%Conditions of human melanocytes in vitro culture

    Institute of Scientific and Technical Information of China (English)

    周梅华; 鲁严

    2011-01-01

    This article summarys safe and effective cultured conditions of human melanocytes in vitro from the basic culture medium, additives, UV and co-culture system.Besides, it sum up the cultured conditions of the hair follicle amelanotic melanocytes and immortalized melanocytes.It is of great significance to the basis research and clinical treatment of the pigment diseases.%从培养人黑素细胞的基本培养液、添加剂、紫外线以及共培养系统等方面综述安全、有效的黑素细胞体外培养条件的研究进展;同时总结了人毛囊无色素性黑素细胞、永生化黑素细胞的培养条件,对色素疾病的基础研究及临床治疗有重要意义.

  9. (Pheo)melanin photosensitizes UVA-induced DNA damage in cultured human melanocytes

    NARCIS (Netherlands)

    Wenczl, E.; Schans, G.P. van der; Roza, L.; Kolb, R.M.; Timmerman, A.J.; Smit, N.P.M.; Pavel, S.; Schothorst, A.A.

    1998-01-01

    The question of whether melanins are photoprotecting and/or photosensitizing in human skin cells continues to be debated. To evaluate the role of melanin upon UVA irradiation, DNA single-strand breaks (ssb) were measured in human melanocytes differing only in the amount of pigment produced by

  10. Isolating highly pure rat spermatogonial stem cells in culture.

    Science.gov (United States)

    Hamra, F Kent; Chapman, Karen M; Wu, Zhuoru; Garbers, David L

    2008-01-01

    Methods are detailed for isolating highly pure populations of spermatogonial stem cells from primary cultures of testis cells prepared from 22- to 24-day-old rats. The procedure is based on the principle that testicular somatic cells bind tightly to plastic and collagen matrices when cultured in serum-containing medium, whereas spermatogonia and spermatocytes do not bind to plastic or collagen when cultured in serum-containing medium. The collagen-non-binding testis cells obtained using these procedures are thus approx. 97% pure spermatogenic cells. Stem spermatogonia are then easily isolated from the purified spermatogenic population during a short incubation step in culture on laminin matrix. The spermatogenic cells that bind to laminin are more than 90% undifferentiated, type A spermatogonia and are greatly enriched in genetically modifiable stem cells that can develop into functional spermatozoa. This method does not require flow cytometry and can also be applied to obtain enriched cultures of mouse spermatogonial stem cells. The isolated spermatogonia provide a highly potent and effective source of stem cells that have been used to initiate in vitro and in vivo culture studies on spermatogenesis.

  11. PURE CULTURE METHOD: GIARDIA LAMBLIA FROM DIFFERENT STOOL SAMPLES

    Directory of Open Access Journals (Sweden)

    H.A YOUSEFI

    2000-03-01

    Full Text Available Introduction. Giardiasis is one of the health problems in the world including Iran. To determine the biochemical and biological problems and also identification of various strains, it is essential to obtain pure culture and then mass production of Giardia lamblia. The goal of this study was to isolate this protozoa purely.
    Methods. Giardia lamblia cysts were isolated from 50 stool samples by use of floating of a four - layer of sucrose method. The cysts were transfered to an inducing solution. Subsequently, they were cultured in a modified culture medium (TYIS-33. Following excystation of trophozoite and its multiplication, the parasite was caltured and purified.
    Findings. Excitation of trophozoite was observed in 40 samples (80 percent from which 22 samples (55 percent yielded pure culture. The doubling time was approximately 13hr and the peak of parasite was observed between third and fourth days.
    Conclusion. The proliferation and growth rate of Giardia lamblia have enabled us to use this method widely. Cystein and ascorbic acid which are present in the induction solution, have a key role in excystation of trophozoite. Purification and passage of samples has facilitated the culture of this parasite in vitro. Therefore this method has yielded better results in comparison with other studies. This is probably due to a decrease in the amount of bovine bile or using different strains of Giardia lamblia in the present study.

  12. Methods of culturing normal human melanocytes in vitro%人正常黑素细胞体外培养方法

    Institute of Scientific and Technical Information of China (English)

    商进; 王震宇

    2015-01-01

    目的:学习人正常黑素细胞的原代培养技术,为进一步研究提供实验模型。方法:分离培养人正常黑素细胞,细胞取材于包皮环切术后的包皮组织;采用左旋多巴染色法进行细胞鉴定。结果:成功培养、传代、纯化了人正常黑素细胞,并经左旋多巴染色证实。结论:动态观察了人正常黑素细胞培养的全过程,验证了细胞的特性,获得了大量纯化的黑素细胞,为进一步的细胞内干预实验提供了物质基础。%Objective:To study the methods of culturing human normal melanocytes, providing experimental models for further investigation. Methods:Cells from prepuce tissue after circumcision were isolated to culture normal human melanocytes which were later identified by Levodopa staining method.Result:Normal human melanocytes were cultured, reproduced and purified successfully, which was identified by Levodopa stai-ning method.Conclusion:The whole culture process of normal human melanocytes has been dynamically observed.The characteristics of melano-cytes have been identified.A large number of purified melanocytes have been obtained, which provides basic materials for further intervention.

  13. Melanogenesis and antioxidant defense system in normal human melanocytes cultured in the presence of chlorpromazine.

    Science.gov (United States)

    Otreba, Michał; Wrześniok, Dorota; Beberok, Artur; Rok, Jakub; Buszman, Ewa

    2015-02-01

    Chlorpromazine is used in the treatment of schizophrenia and psychotic disorders and belongs to phenothiazine class of neuroleptic drugs. It shows severe side effects such as extrapyramidal symptoms as well as ocular and skin disorders, but the mechanism is still not fully established. The aim of this study was to examine the effect of chlorpromazine on cell viability, melanogenesis and antioxidant defense system in normal human melanocytes. It has been demonstrated that chlorpromazine induces concentration dependent loss in cell viability. The value of EC(50) was calculated to be 2.53 μM. Chlorpromazine in lower concentrations (0.0001, 0.001 and 0.01 μM) increased the melanin and microphthalmia-associated transcription factor (MITF) content and tyrosinase activity, while changes of antioxidant enzymes activity were not observed. It suggests that long-term chlorpromazine therapy, even with low drug doses, may lead to hyperpigmentation disorders in skin and/or eye. The use of the analyzed drug in higher concentrations (0.1 and 1.0 μM) caused significant alterations of antioxidant enzymes activity in normal melanocytes, what may explain a potential role of chlorpromazine in the depletion of cellular antioxidant status leading to other adverse effects associated with the high-dose and/or long-term therapy.

  14. Ultra-structural effects of different low-level lasers on normal cultured human melanocytes: an in vitro comparative study.

    Science.gov (United States)

    AlGhamdi, Khalid M; Kumar, Ashok; A Al-Ghamdi, Attieh; Al-Rikabi, Ammar C; Mubarek, Mohammed; Ashour, Abdelkader E

    2016-12-01

    The aim of this study was to investigate the effects of the different types of low-level laser therapy (LLLT) on the ultra-structure and number of melanosomes in normal cultured human melanocytes. Specific effects of various types of LLLT on the ultra-structure of melanosomes have not yet been reported. Melanocytes were exposed to LLLT at an energy level of 2.0 J/cm(2), using a blue (457 nm), red (635 nm), or ultraviolet (UV) (355 nm) laser. After 72 h of irradiation, the melanocytes were fixed in 2.5 % glutaraldehyde (pH 7.2) phosphate buffer for 8 h and analyzed by transmission electron microscopy. Four developmental stages (I to IV) of melanosomes were observed, and their numbers were counted manually. The percentage of stages I, II, III, and IV melanosomes was 12.8, 14.2, 22.6, and 50.3 %, respectively, in the control (sham light). However, the melanosome percentages were 41.2, 5.4, 8.2, and 24.2 % in stages I, II, III, and IV, respectively, in the blue laser-treated group; 58.4, 6.1, 9.3, and 26.2 % for stages I, II, III, and IV, respectively, in the red laser-treated group; and 31.3, 11.1, 16.5, and 41.1 % for stages I, II, III, and IV, respectively, in the UV laser-treated group. The present data show that the amount of stage I is significantly higher (P < 0.0001) in the LLLT-treated cells compared to the control, which indicates significant stimulation of melanogenesis. The red laser was more effective than the other lasers. Moreover, the effects of LLLT on the ultra-structure of melanosomes need to be studied in a larger number of subject groups.

  15. 3D co-cultures of keratinocytes and melanocytes and cytoprotective effects on keratinocytes against reactive oxygen species by insect virus-derived protein microcrystals.

    Science.gov (United States)

    Shimabukuro, Junji; Yamaoka, Ayako; Murata, Ken-Ichi; Kotani, Eiji; Hirano, Tomoko; Nakajima, Yumiko; Matsumoto, Goichi; Mori, Hajime

    2014-09-01

    Stable protein microcrystals called polyhedra are produced by certain insect viruses. Cytokines, such as fibroblast growth factors (FGFs), can be immobilized within polyhedra. Here, we investigated three-dimensional (3D) co-cultures of keratinocytes and melanocytes on collagen gel containing FGF-2 and FGF-7 polyhedra. Melanocytes were observed to reside at the base of the 3D cell culture and melanin was also typically observed in the lower layer. The 3D cell culture model with FGF-2 and FGF-7 polyhedra was a useful in vitro model of the epidermis due to effective melanogenesis, proliferation and differentiation of keratinocytes. FGF-7 polyhedra showed a potent cytoprotective effect when keratinocytes were treated with menadione, which is a generator of reactive oxygen species. The cytoprotective effect was activated by the inositol triphosphate kinase-Akt pathway leading to upregulation of the antioxidant enzymes superoxide dismutase and peroxiredoxin 6.

  16. The presence of embedded bacterial pure cultures in agar plates stimulate the culturability of soil bacteria

    DEFF Research Database (Denmark)

    Burmølle, Mette; Johnsen, Kaare; Abu Al-Soud, Waleed Mohamad Abdel F

    2009-01-01

    Traditional methods for bacterial cultivation recover only a small fraction of bacteria from all sorts of natural environments, and attempts have been made to improve the bacterial culturability. Here we describe the development of a cultivation method, based on the embedment of pure bacterial cu...

  17. Melanocytes--a novel tool to study mitochondrial dysfunction in Duchenne muscular dystrophy.

    Science.gov (United States)

    Pellegrini, Camilla; Zulian, Alessandra; Gualandi, Francesca; Manzati, Elisa; Merlini, Luciano; Michelini, Maria E; Benassi, Luisa; Marmiroli, Sandra; Ferlini, Alessandra; Sabatelli, Patrizia; Bernardi, Paolo; Maraldi, Nadir M

    2013-06-01

    Dystrophin is a subsarcolemmal protein that, by linking the actin cytoskeleton to the extracellular matrix via dystroglycans, is critical for the integrity of muscle fibers. Here, we report that epidermal melanocytes, obtained from conventional skin biopsy, express dystrophin with a restricted localization to the plasma membrane facing the dermal-epidermal junction. In addition the full-length muscle isoform mDp427 was clearly detectable in melanocyte cultures as assessed by immunohistochemistry, RNA, and Western blot analysis. Melanocytes of Duchenne muscular dystrophy (DMD) patients did not express dystrophin, and the ultrastructural analysis revealed typical mitochondrial alterations similar to those occurring in myoblasts from the same patients. Mitochondria of melanocytes from DMD patients readily accumulated tetramethylrhodamine methyl ester, indicating that they are energized irrespective of the presence of dystrophin but, at variance from mitochondria of control donors, depolarized upon the addition of oligomycin, suggesting that they are affected by a latent dysfunction unmasked by inhibition of the ATP synthase. Pure melanocyte cultures can be readily obtained by conventional skin biopsies and may be a feasible and reliable tool alternative to muscle biopsy for functional studies in dystrophinopathies. The mitochondrial dysfunction occurring in DMD melanocytes could represent a promising cellular biomarker for monitoring dystrophinopathies also in response to pharmacological treatments.

  18. Adipose-Derived Stem Cells Improve Efficacy of Melanocyte Transplantation in Animal Skin

    OpenAIRE

    Lim, Won-Suk; Kim, Chang-Hyun; Kim, Ji-Young; Do, Byung-Rok; Kim, Eo Jin; Lee, Ai-Young

    2014-01-01

    Vitiligo is a pigmentary disorder induced by a loss of melanocytes. In addition to replacement of pure melanocytes, cocultures of melanocytes with keratinocytes have been used to improve the repigmentation outcome in vitiligo treatment. We previously identified by in vitro studies, that adipose-derived stem cells (ADSCs) could be a potential substitute for keratinocytes in cocultures with melanocytes. In this study, the efficacy of pigmentation including durability of grafted melanocytes and ...

  19. Differentiating the stem cell pool of human hair follicle outer root sheath into functional melanocytes.

    Science.gov (United States)

    Schneider, Marie; Dieckmann, Christina; Rabe, Katrin; Simon, Jan-Christoph; Savkovic, Vuk

    2014-01-01

    Bench-to-Bedside concepts for regenerative therapy place significant weight on noninvasive approaches, with harvesting of the starting material as a header. This is particularly important in autologous treatments, which use one's bodily constituents for therapy. Precisely the stretch between obtaining therapeutic elements invasively and noninvasively places non-intrusive "sampling" rather than "biopsy" in the center of the road map of developing an autologous regenerative therapy. We focus on such a noninvasively available source of adult stem cells that we carry with us throughout our life, available at our fingertips-or shall we say hair roots, by a simple plucking of hair: the human hair follicle. This chapter describes an explant procedure for cultivating melanocytes differentiated from the stem cell pool of the hair follicle Outer Root Sheath (ORS). In vivo, the most abundant derivatives of the heterogeneous ORS stem cell pool are epidermal cells-melanocytes and keratinocytes which complete their differentiation-either spontaneously or upon picking up regenerative cues from damaged skin-and migrate from the ORS towards the adjacent regenerating area of the epidermis. We have taken advantage of the ORS developmental potential by optimizing explant primary culture, expansion and melanogenic differentiation of resident ORS stem cells towards end-stage melanocytes in order to obtain functional melanocytes noninvasively for the purposes of transplantation and use them for the treatment of depigmentation disorders. Our protocol specifies sampling of hair with their ORS, follicle medium-air interface primary culture, stimulation of cell outgrowth, adherent culture and differentiation of ORS stem cells and precursors towards fully functional melanocytes. Along with cultivation, we describe selection techniques for establishing and maintaining a pure melanocyte population and methods suitable for determining melanocyte identity.

  20. 3D co-cultures of keratinocytes and melanocytes and cytoprotective effects on keratinocytes against reactive oxygen species by insect virus-derived protein microcrystals

    Energy Technology Data Exchange (ETDEWEB)

    Shimabukuro, Junji; Yamaoka, Ayako; Murata, Ken-ichi [Department of Applied Biology, Kyoto Institute of Technology, Kyoto (Japan); Kotani, Eiji [Department of Applied Biology, Kyoto Institute of Technology, Kyoto (Japan); Insect Biomedical Research Center, Kyoto Institute of Technology, Kyoto (Japan); Hirano, Tomoko [Venture Laboratory, Kyoto Institute of Technology, Kyoto (Japan); Nakajima, Yumiko [Functional Genomics Group, COMB, Tropical Biosphere Research Center, University of the Ryukyus, Okinawa (Japan); Matsumoto, Goichi [Division of Oral Surgery, Yokohama Clinical Education Center of Kanagawa Dental University, Yokohama (Japan); Mori, Hajime, E-mail: hmori@kit.ac.jp [Department of Applied Biology, Kyoto Institute of Technology, Kyoto (Japan); Insect Biomedical Research Center, Kyoto Institute of Technology, Kyoto (Japan)

    2014-09-01

    Stable protein microcrystals called polyhedra are produced by certain insect viruses. Cytokines, such as fibroblast growth factors (FGFs), can be immobilized within polyhedra. Here, we investigated three-dimensional (3D) co-cultures of keratinocytes and melanocytes on collagen gel containing FGF-2 and FGF-7 polyhedra. Melanocytes were observed to reside at the base of the 3D cell culture and melanin was also typically observed in the lower layer. The 3D cell culture model with FGF-2 and FGF-7 polyhedra was a useful in vitro model of the epidermis due to effective melanogenesis, proliferation and differentiation of keratinocytes. FGF-7 polyhedra showed a potent cytoprotective effect when keratinocytes were treated with menadione, which is a generator of reactive oxygen species. The cytoprotective effect was activated by the inositol triphosphate kinase–Akt pathway leading to upregulation of the antioxidant enzymes superoxide dismutase and peroxiredoxin 6. - Highlights: • 3D cultures using FGF-2 and FGF-7 microcrystals as a human skin model • Cytoprotection of keratinocytes against ROS by FGF-7 microcrystals • Overexpression of SOD and Prdx6 in keratinocytes by FGF-7 microcrystals.

  1. Blister roof grafting, cultured melanocytes transplantation and non-cultured epidermal cell suspension transplantation in treating stable vitiligo: A mutual self-control study.

    Science.gov (United States)

    Bao, Huaye; Hong, Weisong; Fu, Lifang; Wei, Xiaodong; Qian, Guopei; Xu, Aie

    2015-01-01

    To compare the efficacy of blister roof grafting (BG), cultured melanocytes transplantation (CMT) and non-cultured epidermal cell suspension transplantation (NCES) in the treatment of stable vitiligo. In each person of 83 vitiligo patients one vitiligo macule was selected and divided in three areas for separate treatment with BG, CMT and NCES in the same session. The results were evaluated 12-month post-surgery for the extent of repigmentation and color match. A satisfactory result (>50% repigmentation) was achieved in 92%, 82% and 81% of the 83 patients with the BG, CMT and NCES methods, respectively. Significant differences between the BG and CMT groups (p = 0.038), and between BG and NCES groups (p = 0.017) were observed, but not between the CMT and NCES groups (p = 0.986). The extent of repigmentation on the head neck and trunk was superior to that of the extremities by all the three methods. A difference in the time of onset of repigmentation was observed, with repigmentation first appearing after 10 days, 20-30 days and >30 days in the BG, CMT and NCES groups, respectively. All the three methods are safe and effective to treat vitiligo. Future studies with larger groups are warranted to confirm our results.

  2. Ultrastructural observations and preparation of cultured human epidermal melanocytes' samples%体外培养人表皮黑素细胞样本制备及超微结构观察

    Institute of Scientific and Technical Information of China (English)

    徐丹丹; 陈玮莹; 李伟秋; 方斌; 黄清霞; 梁伟全; 梁彦; 钟耀棠; 王腾彬; 周远军

    2012-01-01

    Purpose To investigate the preparation methods of TEM samples for cultured human epidermal melanocytes and to observe the ultrastructures of the cells. Methods Melanocytes were isolated from children s foreskins and cultured in M254 medium that was supplemented with human melanocyte growth supplement. Purified melanocytes were harvested from culture flask and TEM sample was prepared by conventional method, or cultured on PET film and allowed to attach and then TEM sample was prepared, respectively. Results Transmission electron microscopy showed that the ultrastructure of melanocytes and dendrites in transverse section were observed in samples prepared with conventional method. The intact ultrastructure of melanocytes and their dendrites could be observed in longitudinal section while PET film was applied as cellular support. Conclusions The PET film is an excellent cellular supporting material for the cultivation of human epidermal melanocytes. The ultrastructures of human epidermal melanocytes could be observed in samples prepared in different ways. But the sample that using PET film as cellular support is much beneficial for the ultrastructural observation of metabolism and transferring of melanosome in melanocytes.%目的 探讨体外培养正常人表皮黑素细胞(melanocytes,MC)透射电镜制样方法及超微结构观察的作用.方法 采用M254 MC培养基及添加剂HMGS培养和纯化人表皮MC;分别采用常规制样方法和以PET聚酯薄膜为培养细胞的支持物制备透射电镜样本.结果 人表皮MC透射电镜观察发现,常规制样标本可观察人表皮MC及其树状突起横切面的超微结构.PET聚酯薄膜作为培养MC支持物的样本可观察人表皮MC和树状突起的完整超微结构及树状突起的纵切面超微结构.结论 PET聚酯薄膜可作为人表皮MC体外培养良好的支持材料.应用不同样本制备方法可以观察MC的超微结构,使用PET聚酯薄膜作为细胞支持物制备的标本

  3. The pure-science ideal and democratic culture.

    Science.gov (United States)

    Daniels, G H

    1967-06-30

    These early experiences of pure scientists will have an unmistakable ring of familiarity to anyone familiar with the current situation. Charles Sanders Peirce, with characteristic insight, had stated the fundamental dilemma of the pure scientist operating within a democratic framework. How can one ask the public to provide support, much less facilities, for the intellectual gratification of one select group? A part of the answer, of course, is simply that one cannot. As long as a group is dependent upon public support it must seek some means of contact with the values of the enveloping society, and the moment it does this it departs in some measure from the ideal purity. The schizophrenic attitude described by Dubos therefore became a professional necessity as soon as the new ideal was adopted. Since the time of Gould, scientists have been able to tell each other that the man who based science's claim to support on grounds of immediate practical utility was "no loyal follower and true friend of science" and, at the same time, to trust that the popularizers and technicians would convey a different message to the public. On the whole, they have not been disappointed in their expectation, and there has been little need for them to go beyond the standard formula : Utility is not to be a test of scientific work, but all knowledge will ultimately prove useful. Since the continued existence of scientists in this society depends upon the believability of that vague claim, there is little likelihood hood that the schizophrenia will disappear.

  4. 自体黑素细胞培养移植治疗白癜风的临床研究%Clinical research of autologous melanocytes culture and transplantation for the treatment of vitiligo

    Institute of Scientific and Technical Information of China (English)

    罗卫; 马春林; 吕俊卿; 蔡瑞康

    2012-01-01

    目的 进一步研究和探索黑素细胞培养移植治疗白癜风的方法 和疗效.方法 从发疱壁上获取黑素细胞,行纯黑素细胞培养与增殖,采用移植区刮除种植法行自体黑素细胞培养移植治疗白癜风.结果 本组18例白癜风患者(21块皮损)进行了自体黑素细胞培养移植,总有效率90.48%.结论 此方法较简单,可治疗面积大,治疗后色素分部均匀,值得临床推广和应用.%Objective To explore the method and efficacy of the autologous melanocytes culture and transplantation for the treatment of vitiligo. Methods The melanocytes obtained from suction blisters wall and then culture and proliferation the melanocytes. The vitiliginous lesion is scratched and then transplanted the cultured melanocytes. Results Totally 21 lesions of 18 patients with vitiligo were performed the treatment of autologous melanocytes culture and transplantation, and the total effective rate was 90. 48% . Conclusion It is an optimal approach of autologous melanocytes culture and transplantation for the treatment of vitiligo, which is easy to perform that could cure bigger area with equally distributed pigment. It is worth spreading in clinic.

  5. Comparative study of nickel resistance of pure culture and co-culture of Acidithiobacillus thiooxidans and Leptospirillum ferriphilum.

    Science.gov (United States)

    Xu, Ying; Yin, Huaqun; Jiang, Huidan; Liang, Yili; Guo, Xue; Ma, Liyuan; Xiao, Yunhua; Liu, Xueduan

    2013-09-01

    The effect of Ni²⁺ on the growth and functional gene expression of the pure culture and co-culture of Acidithiobacillus thiooxidans and Leptospirillum ferriphilum has been studied. Compared with the pure culture, the co-culture showed a stronger sulfur and ferrous ion oxidation activity. At 100 mM, A. thiooxidans in co-culture grew faster and had 48 h shorter lag phases. The cell number of A. thiooxidans in co-culture was about 5 times higher than that in pure culture. The existence of A. thiooxidans in co-culture activated the expression of some metal resistance genes in L. ferriphilum at least 16 h in advance. A. thiooxidans in co-culture tends to chose more efficient pathways to transport nickel ion, ensuring the export of heavy metal was faster and more effective than that in pure culture. All the data indicated that there were synergetic interactions between iron- and sulfur-oxidizing bacteria under the stress of Ni²⁺.

  6. 黑素细胞个体化培养体系的建立及应用%Establishment and application of individualized culture system for melanocytes

    Institute of Scientific and Technical Information of China (English)

    洪为松; 傅丽芳; 尉晓冬; 许爱娥

    2011-01-01

    ObjectiveTo establish an individualized culture system for melanocytes, and to estimate its efficacy for the treatment of large-area vitiligo. Methods Hu 16 medium was used for in vitro primary culture of melanocytes isolated from patients with stable segmental vitiligo. Doubling time (DOT), melanin content (M), melanin production (MP) and number of dendrites were examined to evaluate the biological activity of melanocytes. To obtain melanocytes with better biological activity, the components of Hu16 culture medium were adjusted. Ultra pulse CO2 laser was utilized to shave the vitiligous lesions and remove the epidermis followed by autologous transplantation. Follow-up was carried out. Results Melanocytes were obtained from 10 patients with stable segmental vitiligo and cultured. The melanocytes from 6 patients showed relatively short DOT, stable M and MP during the first and seventh passage, and were considered to be at initial or growth stage and applicable to transplantation. The remaining melanocytes from the other 4 patients had displayed long DOT, instable M, MP and dendrite quantity since the third passage; by adjusting the components of culture medium, these cells were induced into growth stage and finally applied to transplantation. A 12-month follow-up revealed that the repigmentation rate was higher than 90% in 7 patients, ranged between 70% and 80% in the remaining 3 patients, with the transplantation area being 116.8 + 75.6 cm2. Conclusions The individualized culture system with adjusted components in culture medium yields melanocytes with satisfying biological activity, which are proved to be effective for the treatment of large-area, segmental and stable vitiligo.%目的 建立有效的黑素细胞个体化培养实验方案,并观察其治疗大面积白癜风的疗效。方法 Hu16培养基体外培养黑素细胞,应用黑素细胞的分裂时间(DOT)、黑素含量(M)、黑素制造量(MP)和树突数作为黑素细胞生物学活

  7. Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes.

    Science.gov (United States)

    Lin, Sung-Jan; Hsiao, Wen-Chu; Jee, Shiou-Hwa; Yu, Hsin-Su; Tsai, Tsen-Fang; Lai, Juin-Yih; Young, Tai-Horng

    2006-10-01

    Though reported limitedly in tissue engineering, modification of cellular functions can be achieved by culturing them into multicellular spheroids. We have shown melanocytes form spheroids on chitosan surface. However, how biomaterials promote spheroid formation has never been systemically investigated. In this work, nylon, which inhibits melanocyte spheroid formation, and chitosan, which promotes melanocyte spheroid formation, are used to prepare nylon/chitosan-blended membranes. Membranes composed of pure nylon, pure chitosan and various ratios of nylon and chitosan are employed to examine their effects on spheroid formation. Melanocytes show better adhesion to nylon membranes than that to chitosan membranes. In blended membranes, as more nylon is incorporated, cell adhesion increases and the trend for spheroid formation decreases. Melanocytes can only form spheroids on membranes with poorer cell adhesion. Examining the surface of the blended membranes shows phase separation of nylon and chitosan. As nylon content increases, the nylon phase on the membrane surface increases and thereby enhances cell adhesion. The opposite trend for cell adhesion and spheroid formation substantiates our hypothesis of spheroid formation on biomaterials: a balance between cell-substrate interaction and cell-cell interaction. The decrease in cell-substrate interaction tilts the balance to a state more favorable for spheroid formation. Our work can serve as a model to investigate the relative strengths of cell-cell and cell-substrate interactions and also pave way to design blended membranes with desired physical properties while preserving the spheroid-forming activity.

  8. FT-IR microspectroscopy in rapid identification of bacteria in pure and mixed culture

    Science.gov (United States)

    Fontoura, Inglid; Belo, Ricardo; Sakane, Kumiko; Cardoso, Maria Angélica Gargione; Khouri, Sônia; Uehara, Mituo; Raniero, Leandro; Martin, Airton A.

    2010-02-01

    In recent years FT-IR microspectroscopy has been developed for microbiology analysis and applied successfully in pure cultures of microorganisms to rapidly identify strains of bacteria, yeasts and fungi. The investigation and characterization of microorganism mixed cultures is also of growing importance, especially in hospitals where it is common to poly-microbial infections. In this work, the rapid identification of bacteria in pure and mixed cultures was studied. The bacteria were obtained from the Institute Oswaldo Cruz culture collection at Brazil. Escherichia coli ATCC 10799 and Staphylococcus aureus ATCC 14456 were analyzed, 3 inoculations were examined in triplicate: Escherichia coli, Staphylococcus aureus and a mixed culture of them. The inoculations were prepared according to McFarland 0.5, incubated at 37 ° C for 6 hours, diluted in saline, placed in the CaF2 window and store for one hour at 50°C to obtain thin film. The measurement was performed by Spectrum Spotlight 400 (Perkin-Elmer) equipment in the range of 4000-900 cm-1, with 32 scans using a transmittance technique with point and image modes. The data were processed (baseline, normalization, calculation of first derivate followed by smoothing with 9 point using a Savitzky-Golay algorithm) and a cluster analysis were done by Ward's algorithm and an excellent discrimination between pure and mixed culture was obtained. Our preliminary results indicate that the FT-IR microspectroscopy associated with cluster analysis can be used to discriminate between pure and mixed culture.

  9. Generation of human melanocytes from induced pluripotent stem cells.

    Directory of Open Access Journals (Sweden)

    Shigeki Ohta

    Full Text Available Epidermal melanocytes play an important role in protecting the skin from UV rays, and their functional impairment results in pigment disorders. Additionally, melanomas are considered to arise from mutations that accumulate in melanocyte stem cells. The mechanisms underlying melanocyte differentiation and the defining characteristics of melanocyte stem cells in humans are, however, largely unknown. In the present study, we set out to generate melanocytes from human iPS cells in vitro, leading to a preliminary investigation of the mechanisms of human melanocyte differentiation. We generated iPS cell lines from human dermal fibroblasts using the Yamanaka factors (SOX2, OCT3/4, and KLF4, with or without c-MYC. These iPS cell lines were subsequently used to form embryoid bodies (EBs and then differentiated into melanocytes via culture supplementation with Wnt3a, SCF, and ET-3. Seven weeks after inducing differentiation, pigmented cells expressing melanocyte markers such as MITF, tyrosinase, SILV, and TYRP1, were detected. Melanosomes were identified in these pigmented cells by electron microscopy, and global gene expression profiling of the pigmented cells showed a high similarity to that of human primary foreskin-derived melanocytes, suggesting the successful generation of melanocytes from iPS cells. This in vitro differentiation system should prove useful for understanding human melanocyte biology and revealing the mechanism of various pigment cell disorders, including melanoma.

  10. Effects of interleukin-6 on cultured human melanocytes in vitro%白介素-6对体外培养的黑素细胞影响的实验研究

    Institute of Scientific and Technical Information of China (English)

    林新瑜; 段西凌; 林鸿刚; 何飞

    2011-01-01

    Objective Establishing the culture of human normal melanocytes in vitro, and studying effects of Interleukin-6 on cultured human melanocytes. Methods MTT method, NaOH 2 assay were employed to measure the cell viability, melanin synthesis of melanocytes, apoptosis rates of melanocytes were detected by using flow cytometer. Results Interleukin-6 induced a mild effect on melanocytic viability, while it showed a significant decrease in the cell counts, melanin synthesis and cell apoptosis. Conclusion Interleukin-6 can inhibit melanocytic proliferation and melanin synthesis. It is worth to study later. It makes valid to make use of Interleukin-6 for treatment of the pigmented skin diseases.%目的 建立正常人表皮黑素细胞体外培养体系,研究白介素-6(Interleukin-6,IL-6)对体外培养的黑素细胞的影响.方法 采用四甲基偶氮唑盐(MTT)法测定白介素-6对黑素细胞活力的影响,NaOH裂解法测定黑素合成,流式细胞仪检测黑素细胞的凋亡率.结果 白介素-6对黑素细胞活力有抑制作用,能使细胞增殖力降低,黑素合成减少,增加黑素细胞的凋亡率.结论 白介素-6对体外培养的黑素细胞增殖及黑素生成都有抑制作用,这为临床应用白介素-6治疗色素性疾病提供了实验依据.

  11. 含己烯雌酚血清对体外培养正常人黑素细胞的作用%Effects of Serum containing Diethylstilbestrol (DES)on cultured human melanocytes in vitro

    Institute of Scientific and Technical Information of China (English)

    龚石; 杨先旭; 张晴; 刘巧

    2012-01-01

    Objective Establishing the culture of human normal melanocytes in vitro,and studying the effects of Serum containing Diethylstilbestrol(DES)on cultured human melanocytes. Methods Viability of melanocytes were measured by MTT method; tyrosinase activity was measured by utilization of L-Dopa as the substrate (Nakajima M method ) and melanocytes melanogenesi by NaOH method. Results Different dosage of Serum containing Diethylstilbestroi (DES) induced a strong promotion on melanocytic viability and tyrosinase activity. Conclusion Serum containing Diethylstilbestrol (DES) can promote melanocytic proliferation and tyrosinase activity. Serum containing Diethylstilbestrol might have some therapenticai effects on correlated pigmented skin diseases .%目的:观察不同浓度含己烯雌酚(DES)血清对体外培养人黑素细胞的增殖和酪氨酸酶活性的影响.方法:采用MTT法测定黑素细胞增殖情况;参考Nakajima M方法测定酪氨酸酶活性;NaOH溶解法测定黑素含量.结果:己烯雌酚血清不同剂量对人黑素细胞增殖及酪氨酸酶活性均有促进作用.结论:己烯雌酚能促进体外培养人黑素细胞增殖及酪氨酸酶活性,对相关皮肤色素性疾病可能有一定的治疗作用.

  12. Wnt inhibitory factor (WIF)-1 promotes melanogenesis in normal human melanocytes.

    Science.gov (United States)

    Park, Tae Jun; Kim, Misun; Kim, Hyeran; Park, Sun Yi; Park, Kyoung-Chan; Ortonne, Jean-Paul; Kang, Hee Young

    2014-01-01

    Wnt signaling plays a role in the differentiation as well as the development of melanocytes. Using a microarray analysis, hyperpigmentary skin of melasma expressed high levels of Wnt inhibitory factor-1 (WIF-1) compared with perilesional normal skin. In this study, the expression and functional roles of WIF-1 on melanocytes were investigated. WIF-1 was expressed both in the melanocytes of normal human skin and in cultured melanocytes. The upregulation of WIF-1 on cultured normal human melanocytes significantly induced expressions of MITF and tyrosinase, which were associated with increased melanin content and tyrosinase activity. Consistent with the stimulatory effect of WIF-1, WIF-1 siRNA reduced melanogenesis in the cells. Moreover, WIF-1 increases pigmentation in melanocytes co-cultured with WIF-1-overexpressed fibroblasts and of organ-cultured human skin. These findings suggest that melanocytes express WIF-1 constitutively in vivo and in vitro and that WIF-1 promotes melanogenesis in normal human melanocytes.

  13. Modulation of the pro-inflammatory cytokines and matrix metalloproteinases production in co-cultivated human keratinocytes and melanocytes.

    Science.gov (United States)

    Decean, H; Perde-Schrepler, M; Tatomir, C; Fischer-Fodor, E; Brie, I; Virag, P

    2013-10-01

    The human epidermis exerts immunoregulatory functions through the variety of cytokines and other molecules elaborated by keratinocytes and melanocytes. Their constitutive production is very low; however, considerably increased upon stimulation. In vivo, keratinocytes and melanocytes have a typical exposure in the skin, referred as melanocyte epidermal unit. In the present study we co-cultivated these cells in vitro proposing to elucidate some communication links in close cell-to-cell association. We assessed the amounts of IL-6, IL-8, and matrix metalloproteinases (MMP-2 and MMP-9) in individually and co-cultured cells, exposed or not to UVB radiation. Normal human epidermal keratinocytes and melanocytes were grown in specific media and supplements. Cells were exposed to UVB radiation (100 mJ/cm(2)) to create comparable stress to the environmental one. Cytokines were determined with ELISA and confirmed with Western blot and metalloproteinases with gel zimography. Pure cultures of keratinocytes and melanocytes released low amounts of cytokines and metalloproteinases, these secretions being enhanced by UVB irradiation. In co-cultures, the cell-to-cell proximity triggered signals which markedly augmented the cytokines' secretions, whereas metalloproteinases were down-regulated. UVB irradiation did not influence either of these secretions in co-cultures. Concurrently with the highest levels of the pro-inflammatory cytokines, MMP-9 was up-regulated creating pro-inflammatory conditions and premises for changes in cellular survival, differentiation and phenotype. A complex network of interactions occurred between keratinocytes and melanocytes in co-cultures, resulting in modulated pro-inflammatory cytokines and metalloproteinases productions. Therefore, any disturbances in the microenvironmental signaling system and its molecular constituents may result in inflammation or even tumorigenesis in the epidermis.

  14. 热处理对体外培养正常人黑素细胞活性的影响%Effect of heat treatment on the viability of cultured normal human melanocytes

    Institute of Scientific and Technical Information of China (English)

    牛建荣; 杨庆琪; 孟如松; 成玉; 赵广

    2011-01-01

    Objective To investigate the effect of heat treatment on the proliferation of, melanin synthesis and tyrosinase activity in cultured normal human melanocytes. Methods Normal human foreskin tissue was obtained by sterile circumcision and melanocytes were harvested by using methods for epidermal cell culture. Basic fibroblast growth factor (bFGF) was utilized as the primary mitogen to establish the culture system of normal human epidermal melanocytes. Masson-Fontana staining was proformed to identify melanocytes.Third-passage melanocytes were treated with hyperthermia at various temperatures (39 ℃, 41 ℃, 42 ℃, 43 ℃ and 45℃) for 1 hour a day for consecutive 3 days followed by the measurement of cell viability with MTT assay. The hyperthermia at optimized temperature was used to treat fourth-passage melanocytes for 1 hour a day for consecutive 3 days; subsequently, the tyrosinase activity were detected with L-Dopa as the substrate, and melanin content was determined in heat-treated and untreated (control) melanocytes. Results The hyperthermia at 42 ℃ exhibited the strongest promotive effect on the proliferation of melanocytes among these 5 hyperthermia conditions. After treatment with hyperthermia at 42 ℃ for 1 hour a day for consecutive 3 days, melanocytes showed an increment in tyrosinase activity by 36.4% and melanin synthesis by 78% compared with the untreated melanocytes (both P<0.05). Conclusions Heat treatment can enhance the proliferation of cultured human melanocytes, promote their melanin synthesis and tyrosinase activity.%目的 探讨热处理对体外培养的正常人黑素细胞的增殖活性、黑素合成及其酪氨酸酶活性的影响.方法 无菌操作获取正常人包皮环切术后的包皮,按照皮肤表皮细胞培养法获取黑素细胞,利用碱性成纤维细胞生长因子(bFGF)为主要有丝分裂原建立正常人表皮黑素细胞培养系,采用Masson-Fontana染色法对培养细胞进行鉴定,确定细胞种系,

  15. Comparative evaluation of the hydrogen production by mixed consortium, synthetic co-culture and pure culture using distillery effluent.

    Science.gov (United States)

    Mishra, Preeti; Roy, Shantonu; Das, Debabrata

    2015-12-01

    Wastewater comprises of various carbon sources. So, the use of microbial consortium may improve the hydrogen production and organic reduction. The present study deals with biohydrogen production by acidogenic mixed consortia (AMC), synthetic co-culture (Klebsiella pneumoniae IIT-BT 08 and Citrobacter freundii IIT-BT L139) and pure culture using distillery effluent (DE). Higher hydrogen yield was observed in case of AMC (9.17 mol/kg CODreduced) as compared to the synthetic co-culture and pure culture. PCR-DGGE analysis indicated that the consortium was predominated by species closely affiliated to Clostridium sp. The average hydrogen production rate was 267 mL/Lh. The maximum hydrogen production rate (Rm), hydrogen production potential (P) and lag time (λ) by AMC using DE were 507.2 mL/Lh, 3729 m/L and 2.04 h, respectively. Maximum gaseous energy recovery by AMC was found to be higher by 21.9% and 45.4% than that of using co-culture and pure culture respectively.

  16. METHOD FOR INDUCING SPORULATION OF PURE CULTURES OF THE MYXOMYCETE PHYSARUM POLYCEPHALUM

    Science.gov (United States)

    Daniel, John W.; Rusch, Harold P.

    1962-01-01

    Daniel, John W. (University of Wisconsin, Madison) and Harold P. Rusch. Method for inducing sporulation of pure cultures of the myxomycete Physarum polycephalum. J. Bacteriol. 83:234–240. 1962.—Techniques for inducing sporulation of pure cultures of Physarum polycephalum are described. Small plasmodia grown in agitated culture were harvested, allowed to fuse into large plasmodia (1.3 g wet weight each), and, after incubation in the dark on a salts medium, exposed to light for 2 hr. The ensuing formation of sporangia containing spores was complete after 12 to 16 hr. The obligatory conditions for sporulation are: (i) an optimal growth age occurring just prior to the maximal growth of the organism and at a time when nutrients in the medium are exhausted; (ii) 4 days of incubation on a medium containing only inorganic salts and niacin or tryptophan; and (iii) subsequent illumination with light of wavelengths between 350 and 500 mμ. PMID:13883384

  17. 同种异体淋巴细胞与黑素细胞混合培养的实验研究%Allogeneic Mixed Melanocyte-Lymphocyte Culture: An Experimental Study

    Institute of Scientific and Technical Information of China (English)

    项蕾红; 郑志忠; 祝绿川; 廖康煌; 傅雯雯

    2001-01-01

    Objective To investigate the melanocyte-lymphocyte reacion when the allogeneic lymphocytes are cultured with normal melanocytes. Methods 3H - thymidine was incorporated into the mixed culture and the transformation and proliferation rates of lymphocytes were detected by liquid scintillation counting and expressed as cpm. Electron microscopy was used to observe the ultrastructure of melanocytes after mixed culture.Results The results of lymphocyte proliferation were expressed by the stimulation indexes. The stimulation indexes in active vitiligo group was significantly different from that in stable vitiligo group and normal controls. The stimulation indexes of the melanocyte-stimulated group was significantly different from that of ConA-stimulated group. In the mixed melanocyte-lymphocyte reaction, the allogeneic lymphocytes had little effect on the melanocytes. The ultrastructure of the melanocytes in the mixed culture showed normal morphology and normal function of synthesis of melanin. Conclusion As a specific antigen in mixed melanocyte-lymphocyte reaction, melanocyte has a weak effect on the lymphocytes. The melanocytes from stable stage vitiligo patients seem more suitble to be allografted.%目的黑素细胞与同种异体淋巴细胞混合培养,从体外实验来研究排斥反应的可能性。方法采用3H-dTR掺入同位素液态闪烁计数法测定淋巴细胞的转化增殖率,并用透射电镜观察混合培养后黑素细胞的超微结构。结果黑素细胞对同种异体淋巴细胞的转化增殖率与ConA刺激淋巴细胞转化增殖的阳性对照比较,结果显示黑素细胞的促淋巴细胞转化增殖的特异性抗原作用较弱。进一步比较黑素细胞对不同病期白癜风患者淋巴细胞的影响,发现黑素细胞对活动期白癜风患者淋巴细胞的刺激作用相对较强,而稳定期患者和正常人对照组结果差异无显著性。同时电镜观察结果显示同种异体的淋巴细胞对黑

  18. rhCSF3 accelerates the proliferation of human melanocytes in culture through binding CSF3R and the expression of CSF3R transcripts.

    Science.gov (United States)

    Lu, Yan; Guo, Ze; Zhou, Mei-Hua; Li, Xue; Sun, Jie; Gong, Qing-Li; Zhu, Wen-Yuan

    2015-05-01

    Melanogenic paracrine and autocrine cytokine networks have recently been discovered in vitro between melanocytes and other types of skin cells. Granulocyte colony-stimulating factor receptor (CSF3R) controls the survival, proliferation and differentiation of many kinds of cells, including neutrophils. To understand the function of CSF3R and recombinant human granulocyte colony-stimulating factor (rhCSF3) on melanocyte proliferation, this study compared the expression of CSF3R and the effects of rhCSF3 in primary human melanocytes, neutrophils and HEL 92.1.7 cells. The results show that CSF3R is localized in the cytoplasm and on cell membranes of melanocytes and neutrophils. The percentage of CSF3R(+) melanocytes was higher than CSF3R(+) HEL 92.1.7 cells, but was lower than CSF3R(+) neutrophils. Both CSF3R mRNA and CSF3R protein levels in melanocytes were higher than in HEL 92.1.7 cells, but were lower than in neutrophils. Treatment with rhCSF3 increased the proliferation of human melanocytes, but not their tyrosinase activity. Transcripts of CSF3R in human melanocytes, M14, A375 melanoma and A431 squamous cell carcinoma cells were also detected. Expression of the CSF3R V3 transcript was lower in melanocytes than in M14, A375 melanoma and A431 squamous cell carcinoma cells. In conclusion, rhCSF3 can promote melanocyte proliferation through CSF3R without affecting tyrosinase activity.

  19. 以人羊膜为支架的表皮黑素细胞培养与生物活性鉴定%Culture of human epidermal melanocytes with human amniotic membrane as a scaffold and their biological ability identification

    Institute of Scientific and Technical Information of China (English)

    佘小光; 刘小明; 雷铁池

    2011-01-01

    目的 从吸引疱疱顶微小皮片建立人表皮黑素细胞(MC)原代培养,以人羊膜为支架培养MC,观察羊膜对MC黏附、增殖和树突发育的影响.方法 用负压吸疱法分离疱顶微小皮片,多巴染色后对取自同一个体下腹部和前臂屈侧皮片的MC进行计数.皮片经胰酶消化后自基底面轻刮表皮细胞建立原代MC培养,将培养的第2~4代MC接种至新鲜或冻存羊膜上共孵育不同时间后,倒置显微镜下观察MC形态和树突发育.行石蜡包埋组织切片,HE染色后观察MC对羊膜的黏附.用MTT比色法测定接种在羊膜上的MC存活率.结果 前臂屈侧吸引疱疱顶皮片多巴染色阳性细胞个数为(1543.1±113.3)个/mm2,腹部皮片为(857.4±101.7)个/mm2.在4周内建立MC原代培养至少需要从2个疱顶(面积约25.1 mm2)的前臂皮片获取的表皮细胞.倒置显微镜下观察,与单纯接种细胞培养皿的MC比较,接种至新鲜或冻存羊膜上继续培养4、8、12 d的MC形态多为两极,树突细长延伸.HE染色示MC能黏附并均匀分布在羊膜的基底膜层表面.MTT法测定结果示,羊膜对MC的分裂增殖有抑制,但冻存羊膜与新鲜羊膜对MC增殖抑制差异无统计学意义(P>0.05).结论 人前臂屈侧皮肤富含活性MC,是获取吸引疱疱顶微小皮片建立MC原代培养的较好部位.人羊膜支持MC黏附生长和树突发育,有望成为一种能荷载体外培养扩增MC的人源生物支架.%Objective To establish a primary culture of human melanocytes from tiny skin sheets harvested by using a suction blister method, to carry out a serial subcultivation of the melanocytes with human amniotic membrane (AM) as a scaffold, and to observe the influence of AM on the adhesion, proliferation and dendrite development of melanocytes. Methods Tiny skin sheets were collected from the flexual forearm or lower abdomen of a healthy male volunteer by a suction blister method and melanocytes in the skin sheet were

  20. Bioelectrochemical reduction of carbon dioxide by pure culture at the cathode

    DEFF Research Database (Denmark)

    Aryal, Nabin; Tremblay, Pier-Luc; Chen, Leifeng

    2014-01-01

    Microbial electrosynthesis (MES) is an innovative approach in which microbes use electricity toreduce carbon dioxide and produce chemical commodities. This process relies on the ability of electroautotrophic microbes to accept electron from an electrode. The concept of MES has already been...... demonstrated with pure cultures of acetogenic bacteria such as Sporomusa ovate DSM-2662 and Clostridium ljungdahlii. Until now, electron transfer rates from the cathode to the bestelectroautotroph, S. ovata, are still orders of magnitude lower than what is observed in bioanodicprocesses with electrigenic...... bacteria. Hence, we are screening other pure cultures for better MES activities. These bacterial species were pre-selected based on several criteria such as their presence in enrichments of environmental samples in MES systems, their capacity to fix CO2, their incapacity to sporulate, and their ability...

  1. Hydrogen isotope fractionation by Methanothermobacter thermoautotrophicus in coculture and pure culture conditions

    Science.gov (United States)

    Yoshioka, Hideyoshi; Sakata, Susumu; Kamagata, Yoichi

    2008-06-01

    We grew a hydrogen-utilizing methanogen, Methanothermobacter thermoautotrophicus strain ΔH, in coculture and pure culture conditions to evaluate the hydrogen isotope fractionation associated with carbonate reduction under low (6 mM; pure culture) concentrations of H 2 in the headspace. In the cocultures, which were grown at 55 °C with a thermophilic butyrate-oxidizing syntroph, the hydrogen isotopic relationship between methane and water was well represented by the following equation: δD=0.725(±0.003)·δDO-275(±3), in which the hydrogen isotope fractionation factor ( αH) was 0.725 ± 0.003. The relationship was consistent with the isotopic data on methane and water from terrestrial fields (a peat bog in Washington State, USA, and a sandy aquifer in Denmark), where carbonate reduction was reported to be the dominant pathway of methanogenesis. In the pure cultures, grown at 55 and 65 °C, the αH values were 0.755 ± 0.014 and 0.749 ± 0.014, respectively. Dependence of αH on growth temperature was not observed. The αH value at 55 °C in the pure culture was slightly higher than that in the coculture, a finding that disagrees with a hypothesis proposed by Burke [Burke, Jr. R. A. (1993) Possible influence of hydrogen concentration on microbial methane stable hydrogen isotopic composition. Chemosphere26, 55-67] that hydrogen isotope fractionation between methane and water increases (and αH decreases) with increasing H 2 concentration.

  2. Application of the Mixture Design to Design the Formulation of Pure Cultures in Tibetan kefir

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    To obtain the optimization formulation of pure cultures in Tibetan kefir, the influence of the different mixtures of five strains in the pure cultures in Tibetan kefir on the flavor components in fermented milk was studied using the mixture design. The regression model on microorganism composition and main metabolites was established. The results suggested that the predictable production of lactate reached the maximum of 8.16 g L-1, while the most predictable production of diacetyl, ethanol, and CO2 were 77.23 mg L-1, 4259 mg L-1, and 2.12 g L-1, respectively. Based on these, the response values that satisfied all expectations were optimized, and the most excellent combination was Lactococcus lactis 27%, Leuconostoc mesenteroides 37%, Lactobacillus kefiri 11%, Lactobacillus casei 10%, and Kluyveromyces marxianus 15%. With the aid of analysis software (Design-expert 6.0.5), the formulation of pure cultures in Tibetan kefir can be optimized for several responses and the best formulation can be obtained.

  3. Development of a PCR test for identification of Haemophilus somnus in pure and mixed cultures

    DEFF Research Database (Denmark)

    Angen, Øystein; Ahrens, Peter; Tegtmeier, Conny

    1998-01-01

    a single colony of H. somnus in the presence of 10(9) CFU of P. multocida even after 2 days of incubation. In conclusion, the present PCR test has been shown to represent a specific test for identification of H. somnus both in pure and mixed cultures. It represents a quick, sensitive and reliable method...... rise to an amplicon in the PCR test. The performance of the test on mixed cultures was evaluated by adding P. multocida to serial dilutions of H. somnus and incubating the agarplates for 1 and 2 days. This showed that the PCR test applied to the harvest from an agarplate can be expected to detect...

  4. Algal recycling enhances algal productivity and settleability in Pediastrum boryanum pure cultures.

    Science.gov (United States)

    Park, Jason B K; Craggs, Rupert J; Shilton, Andy N

    2015-12-15

    Recycling a portion of gravity harvested algae (i.e. algae and associated bacteria biomass) has been shown to improve both algal biomass productivity and harvest efficiency by maintaining the dominance of a rapidly-settleable colonial alga, Pediastrum boryanum in both pilot-scale wastewater treatment High Rate Algal Ponds (HRAP) and outdoor mesocosms. While algal recycling did not change the relative proportions of algae and bacteria in the HRAP culture, the contribution of the wastewater bacteria to the improved algal biomass productivity and settleability with the recycling was not certain and still required investigation. P. boryanum was therefore isolated from the HRAP and grown in pure culture on synthetic wastewater growth media under laboratory conditions. The influence of recycling on the productivity and settleability of the pure P. boryanum culture was then determined without wastewater bacteria present. Six 1 L P. boryanum cultures were grown over 30 days in a laboratory growth chamber simulating New Zealand summer conditions either with (Pr) or without (Pc) recycling of 10% of gravity harvested algae. The cultures with recycling (Pr) had higher algal productivity than the controls (Pc) when the cultures were operated at both 4 and 3 d hydraulic retention times by 11% and 38% respectively. Furthermore, algal recycling also improved 1 h settleability from ∼60% to ∼85% by increasing the average P. boryanum colony size due to the extended mean cell residence time and promoted formation of large algal bio-flocs (>500 μm diameter). These results demonstrate that the presence of wastewater bacteria was not necessary to improve algal productivity and settleability with algal recycling.

  5. Fermentation of residual glycerol by Clostridium acetobutylicum ATCC 824 in pure and mixed cultures.

    Science.gov (United States)

    Dams, Rosemeri I; Guilherme, Alexandre A; Vale, Maria S; Nunes, Vanja F; Leitão, Renato C; Santaella, Sandra T

    2016-12-01

    The aim of this research was to estimate the production of hydrogen, organic acids and alcohols by the strain of Clostridium acetobutylicum ATCC 824 using residual glycerol as a carbon source. The experiments were carried out in pure and mixed cultures in batch experiments. Three different sources of inocula for mixed culture were used. Ruminal liquid from goats and sludge collected from two upflow anaerobic sludge blanket reactors treating municipal wastewater and brewery effluent were tested for hydrogen, organic acids and alcohols production with or without C. acetobutylicum ATCC 824. The main detected end-products from the glycerol fermentation were hydrogen, organic acids (acetic, propionic, butyric and caproic) and alcohol (ethanol and 1,3-propanediol - 1,3PD). High hydrogen (0.44 mol H2/mol glycerol consumed) and 1,3PD (0.32 mol 1,3PD/mol glycerol consumed) yields were obtained when the strain C. acetobutylicum ATCC 824 was bioaugmented into the sludge from municipal wastewater using 5 g/L of glycerol. Significant concentrations of n-caproic acid were detected in the ruminal liquid when amended with C. acetobutylicum ATCC 824. The results suggest that glycerol can be used for the generation of H2, 1,3PD and n-caproic acid using C. acetobutylicum ATCC 824 as agent in pure or mixed cultures.

  6. 氧化应激增强体外培养人黑素细胞异位表达免疫活性分子%Ectopic Expressions of Immunological Markers on Cultured Human Melanocytes Are Increased by Oxidative Stress

    Institute of Scientific and Technical Information of China (English)

    罗龙飞; 刘小明; 雷铁池

    2012-01-01

    目的:比较观察氧化应激状态下正常人表皮黑素细胞(MCs)与白癜风非皮损区MCs细胞膜表达CD40,CD80,HIA-DR和ICAM免疫分子水平的变化.方法:自5例健康成年包皮和5例稳定期白癜风下腹部非皮损区分别建立正常人表皮MCs和白癜风MCs的原代培养;将培养第2-3代细胞分别用100 mmol/L过氧化氢(H2O2)作用1h或1.5 J/cm2长波紫外线(UVA)照射进行氧化应激处理;用二氯荧光素标记法测定两种MCs在H2O2处理和UVA照射前后胞内活性氧基(ROS)水平变化;用流式细胞仪分析CD40,CD80,HLA-DR和ICAM 4种免疫分子表达水平变化;用分光光度计法测定两种MCs过氧化氢酶活性.结果:过氧化氢酶活性测定结果显示白癜风MCs过氧化氢酶活性较正常人表皮MC明显减低(P<0.01).细胞内ROS水平测定发现H2O2处理和UVA照射均导致白癜风非皮损区MCs细胞内ROS水平急剧增高,与正常人MCs比较ROS的荧光强度有显著统计学差异(P<0.01).流式细胞仪检测结果表明除HLA-DR外,UVA照射可诱导白癜风非皮损区MCs高表达CD40,CD80和ICAM1,与未照射组白癜风MCs和UVA照射组正常人MCs比较均有显著统计学差异(P<0.01).结论:白癜风非皮损区MCs的抗氧化能力明显减低.UVA照射能上调白癜风MCs胞膜异位表达CD40,CD80和ICAM1免疫分子,提示在氧化应激状态下白癜风MCs可兼有增强的抗原呈递细胞样功能.%Objective: To analyze the changes in ectopic expressions of immunological markers on cultured human melanocytes from healthy subjects and stable vitiligo patients in the presence or absence of oxidative stress. Methods; Cultured human melanocytes were isolated from the foreskins of five healthy subjects and the uninvolved abdominal skins of five stable vitiligo patients. The cells at the second to third passages were subsequently exposed to 100 mmol/L H2O2 for 1 h or 1. 5 J/cm2 UVA radiation. The levels of intracellular reactive oxygen species

  7. Optimization of Differential Display of Prokaryotic mRNA: Application to Pure Culture and Soil Microcosms

    OpenAIRE

    Fleming, James T.; Yao, Wen-Hsiang; Sayler, Gary S.

    1998-01-01

    The differential display (DD) technique, which is widely used almost exclusively for eukaryotic gene discovery, was optimized to detect differential mRNA transcription from both pure-culture and soil-derived bacterial RNA. A model system which included toluene induction of todC1 in Pseudomonas putida F1 was used to optimize the procedure. At 24-h tod induction was determined to be approximately 8 × 107 transcripts/μg or 0.08% of the total mRNA. The primer concentration, primer length, anneali...

  8. Selective proliferation of normal human melanocytes in vitro in the presence of phorbol ester and cholera toxin.

    OpenAIRE

    Eisinger, M.; Marko, O

    1982-01-01

    Cultures consisting almost entirely of human melanocytes were obtained from epidermal single-cell suspensions by using phorbol 12-myristate 13-acetate (10 ng/ml) in the culture medium. At this concentration, phorbol ester is toxic to human keratinocytes but not to melanocytes. When the seeding density was optimal (0.8-2 x 10(4)/cm2) and the medium contained both phorbol ester and cholera toxin, melanocytes proliferated extensively. Under these conditions, human melanocytes could be passaged s...

  9. Leaching of pyrite by acidophilic heterotrophic iron-oxidizing bacteria in pure and mixed cultures

    Energy Technology Data Exchange (ETDEWEB)

    Bacelar-Nicolau, P.; Johnson, D.B. [Univ. of Wales, Bangor (United Kingdom). School of Biological Sciences

    1999-02-01

    Seven strains of heterotrophic iron-oxidizing acidophilic bacteria were examined to determine their abilities to promote oxidative dissolution of pyrite (FeS{sub 2}) when they were grown in pure cultures and in mixed cultures with sulfur-oxidizing Thiobacillus spp. Only one of the isolates (strain T-24) oxidized pyrite when it was grown in pyrite-basal salts medium. However, when pyrite-containing cultures were supplemented with 0.02% (wt/vol) yeast extract, most of the isolates oxidized pyrite, and one (strain T-24) promoted rates of mineral dissolution similar to the rates observed with the iron-oxidizing autotroph Thiobacillus ferroxidans. Pyrite oxidation by another isolate (strain T-21) occurred in cultures containing between 0.005 and 0.05% (wt/vol) yeast extract but was completely inhibited in cultures containing 0.5% yeast extract. Ferrous iron was also needed for mineral dissolution by the iron-oxidizing heterotrophs, indicating that these organisms oxidize pyrite via the indirect mechanism. Mixed cultures of three isolates (strains T-21, T-232, and T-24) and the sulfur-oxidizing autotroph Thiobacillus thiooxidans promoted pyrite dissolution; since neither strains T-21 and T-23 nor T. thiooxidans could oxidize this mineral in yeast extract-free media, this was a novel example of bacterial synergism. Mixed cultures of strains T-21 and T-23 and the sulfur-oxidizing mixotroph Thiobacillus acidophilus also oxidized pyrite but to a lesser extent than did mixed cultures containing T. thiooxidans. Pyrite leaching by strain T -23 grown in an organic compound-rich medium and incubated either shaken or unshaken was also assessed. The potential environmental significance of iron-oxidizing heterotrophs in accelerating pyrite oxidation is discussed.

  10. Transplantation of individualized cultured autologous melanocytes for the treatment of vitiligo%个体化培养自体黑素细胞移植治疗白癜风

    Institute of Scientific and Technical Information of China (English)

    张迪敏; 洪为松; 傅丽芳; 钱国培; 许爱娥

    2010-01-01

    目的 探讨使用个体化培养基进行自体黑素细胞培养移植治疗白癜风的疗效.方法 负压吸疱获取患者正常表皮片,制成细胞悬液,在Hu16黑素细胞选择性培养基中培养.检测黑素细胞分裂时间(DOT)和黑素含量,根据DOT的大小、黑素含量和细胞形态,调整血清、细胞因子浓度及补充内皮素-1,进行个体化黑素细胞培养.经2~5次传代后收集黑素细胞,白斑区用超脉冲CO2激光磨削后进行黑素细胞移植,随访观察复色效果.结果 共治疗155例稳定期白癜风患者的204处皮损,进行1次移植119例,进行2~4次移植36例.应用个体化黑素细胞培养后细胞扩增可达50~80倍.84.80%的皮损复色面积超过50%,其中52.94%的皮损复色画积超过90%,且复色均匀,未见瘢痕及其他不良反应.性别、年龄、病程长短和皮损面积大小对疗效没有影响.节段型白癜风移植疗效好于寻常型白癜风,两组有效率分别为93.62%和82.16%,痊愈率分别为65.96%和49.04%.手臂和腿部的皮损(不包括肘部和膝盖)移植后痊愈率达73.08%,疗效好于躯干、面颈;肢端皮损疗效最差,痊愈率仅为25.93%.结论 个体化培养技术能提高白癜风患者黑素细胞的培养成功率与细胞扩增倍数.体外培养的自体黑素细胞移植治疗稳定期白癜风疗效肯定,用少量供皮区即可治疗大面积皮损,值得临床应用.%Objective To evaluate the therapeutic effect of transplantation of autologous melanocytes cultured with individualized medium in vitiligo. Methods Donor skin was obtained by suction blisters from a normally pigmented area of the abdomen of 155 patients with vitiligo. The roof of the blisters was clipped and digested with trypsin, then the suspension of epidermal cells and melanocytes were cultured in Hu16 medium.The cell division time (DOT) and melanin content of cultured melanocytes were measured followed by the adjustment of concentration of fetal calf

  11. Isolation and Initial Characterization of A Pure Cultures Capable to Degradation Methyl tert- Butyl Ether (MTBE

    Directory of Open Access Journals (Sweden)

    A Nikpey

    2006-07-01

    Full Text Available Methyl tert-butyl ether (MTBE, a gasoline octane enhancer, was introduced as a substitute for lead tetraethyl over 30 years ago. Widespread use of MTBE in gasoline, has introduced MTBE into the environment compartments, mostly into the under ground and surface water and water as a second most frequently detected contaminant. In this study, we have isolated pure cultures from bacterial consortium capable to use MTBE as a sole carbon and energy source. MTBE biodegradation rate was measured in headspace by gas chromatography. Initial liner rates of biodegradation by Pinpoint and white strains were found 2.9 mg and 3 mg MTBE hˉ1 gˉ1 wet biomass, respectively. The results of 16S rDNA PCR disclosed similarities in the banding patterns between the cultures, and the known degrading strain PM1. The results of this study suggest promising perspectives for engineering the in situ bioremediation of MTBE.

  12. A serendipic legacy: Erwin Esmarch's isolation of the first photosynthetic bacterium in pure culture.

    Science.gov (United States)

    Gest, H

    1995-01-01

    During the 1880's, Erwin von Esmarch was a junior associate ('Assistent') of Robert Koch studying bacteria of medical significance. In 1887, he isolated the first example of spiral-shaped bacteria in pure culture, from the dry residue of a dead mouse that he had suspended sometime earlier in Berlin tap-water. Under certain conditions, colonies of the organism were the color of red wine, and this led Esmarch to name the bacterium Spirillum rubrum. Twenty years later, Hans Molisch demonstrated that S. rubrum, an apparent heterotroph, was in fact a non-oxygenic purple photosynthetic bacterium, and it was renamed Rhodospirillum rubrum. Esmarch was a careful investigator and his classic paper of 1887 details the serendipitous isolation and general characteristics of the first pure culture of an anoxyphototroph, which later played a prominent role as an experimental system for study of basic aspects of bacterial photosynthesis. This report includes an English translation of his original paper (in German), a commentary on the historical significance of 'Esmarch's spirillum', and a summary of Esmarch's career.

  13. Production and optimization of L-glutaminase enzyme from Hypocrea jecorina pure culture.

    Science.gov (United States)

    Bülbül, Dilara; Karakuş, Emine

    2013-01-01

    L-Glutaminase (L-glutamine amidohydrolase, EC 3.5.1.2) is the important enzyme that catalyzes the deamination of L-glutamine to L-glutamic acid and ammonium ions. Recently, L-glutaminase has received much attention with respect to its therapeutic and industrial applications. It acts as a potent antileukemic agent and shows flavor-enhancing capacity in the production of fermented foods. Glutaminase activity is widely distributed in plants, animal tissues, and microorganisms, including bacteria, yeasts, and fungi. This study presents microbial production of glutaminase enzyme from Hypocrea jecorina pure culture and determination of optimum conditions and calculation of kinetic parameters of the produced enzyme. The optimum values were determined by using sa Nesslerization reaction for our produced glutaminase enzyme. The optimum pH value was determined as 8.0 and optimum temperature as 50°C for the glutaminase enzyme. The Km and Vmax values, the kinetic parameters, of enzyme produced from Hypocrea jecorina, pure culture were determined as 0.491 mM for Km and 13.86 U/L for Vmax by plotted Lineweaver-Burk graphing, respectively. The glutaminase enzyme from H. jecorina microorganism has very high thermal and storage stability.

  14. Production of Fusaric Acid by Fusarium spp. in Pure Culture and in Solid Medium Co-Cultures

    Directory of Open Access Journals (Sweden)

    Nadine Bohni

    2016-03-01

    Full Text Available The ability of fungi isolated from nails of patients suffering from onychomycosis to induce de novo production of bioactive compounds in co-culture was examined. Comparison between the metabolite profiles produced by Sarocladium strictum, by Fusarium oxysporum, and by these two species in co-culture revealed de novo induction of fusaric acid based on HRMS. Structure confirmation of this toxin, using sensitive microflow NMR, required only three 9-cm Petri dishes of fungal culture. A targeted metabolomics study based on UHPLC-HRMS confirmed that the production of fusaric acid was strain-dependent. Furthermore, the detected toxin levels suggested that onychomycosis-associated fungal strains of the F. oxysporum and F. fujikuroi species complexes are much more frequently producing fusaric acid, and in higher amount, than strains of the F. solani species complex. Fusarium strains producing no significant amounts of this compound in pure culture, were shown to de novo produce that compound when grown in co-culture. The role of fusaric acid in fungal virulence and defense is discussed.

  15. Production of Fusaric Acid by Fusarium spp. in Pure Culture and in Solid Medium Co-Cultures.

    Science.gov (United States)

    Bohni, Nadine; Hofstetter, Valérie; Gindro, Katia; Buyck, Bart; Schumpp, Olivier; Bertrand, Samuel; Monod, Michel; Wolfender, Jean-Luc

    2016-03-18

    The ability of fungi isolated from nails of patients suffering from onychomycosis to induce de novo production of bioactive compounds in co-culture was examined. Comparison between the metabolite profiles produced by Sarocladium strictum, by Fusarium oxysporum, and by these two species in co-culture revealed de novo induction of fusaric acid based on HRMS. Structure confirmation of this toxin, using sensitive microflow NMR, required only three 9-cm Petri dishes of fungal culture. A targeted metabolomics study based on UHPLC-HRMS confirmed that the production of fusaric acid was strain-dependent. Furthermore, the detected toxin levels suggested that onychomycosis-associated fungal strains of the F. oxysporum and F. fujikuroi species complexes are much more frequently producing fusaric acid, and in higher amount, than strains of the F. solani species complex. Fusarium strains producing no significant amounts of this compound in pure culture, were shown to de novo produce that compound when grown in co-culture. The role of fusaric acid in fungal virulence and defense is discussed.

  16. Effects of Single Herbal Drugs on Adhesion and Migration of Melanocytes

    Institute of Scientific and Technical Information of China (English)

    Zhang Xianqi; Feng Jie; Mu Kuanhou; Ma Huiqun; Niu Xinwu; Liu Chao; Dang Qianli

    2005-01-01

    Objective: To find herbs with effects on adhesion and migration of melanocytes in vitro. Materials and Methods: Ethanol extracts from 14 herbs were tested. Normal human melanocytes were obtained from neonatal foreskin, and the 48-well culture dish covered with fibronectin was used for the melanocyte adhesion assay. Motility was assessed by using the micropore filter method. Results: The extracts of could promote both adhesion and migration of melanocytes. Conclusion: The above herbs may play a role through promoting adhesion and/or migration of melanocytes in the treatment of vitiligo.

  17. Mineral and iron oxidation at low temperatures by pure and mixed cultures of acidophilic microorganisms.

    Science.gov (United States)

    Dopson, Mark; Halinen, Anna-Kaisa; Rahunen, Nelli; Ozkaya, Bestamin; Sahinkaya, Erkan; Kaksonen, Anna H; Lindström, E Börje; Puhakka, Jaakko A

    2007-08-01

    An enrichment culture from a boreal sulfide mine environment containing a low-grade polymetallic ore was tested in column bioreactors for simulation of low temperature heap leaching. PCR-denaturing gradient gel electrophoresis and 16S rRNA gene sequencing revealed the enrichment culture contained an Acidithiobacillus ferrooxidans strain with high 16S rRNA gene similarity to the psychrotolerant strain SS3 and a mesophilic Leptospirillum ferrooxidans strain. As the mixed culture contained a strain that was within a clade with SS3, we used the SS3 pure culture to compare leaching rates with the At. ferrooxidans type strain in stirred tank reactors for mineral sulfide dissolution at various temperatures. The psychrotolerant strain SS3 catalyzed pyrite, pyrite/arsenopyrite, and chalcopyrite concentrate leaching. The rates were lower at 5 degrees C than at 30 degrees C, despite that all the available iron was in the oxidized form in the presence of At. ferrooxidans SS3. This suggests that although efficient At. ferrooxidans SS3 mediated biological oxidation of ferrous iron occurred, chemical oxidation of the sulfide minerals by ferric iron was rate limiting. In the column reactors, the leaching rates were much less affected by low temperatures than in the stirred tank reactors. A factor for the relatively high rates of mineral oxidation at 7 degrees C is that ferric iron remained in the soluble phase whereas, at 21 degrees C the ferric iron precipitated. Temperature gradient analysis of ferrous iron oxidation by this enrichment culture demonstrated two temperature optima for ferrous iron oxidation and that the mixed culture was capable of ferrous iron oxidation at 5 degrees C.

  18. Optical cell separation from three-dimensional environment in photodegradable hydrogels for pure culture techniques.

    Science.gov (United States)

    Tamura, Masato; Yanagawa, Fumiki; Sugiura, Shinji; Takagi, Toshiyuki; Sumaru, Kimio; Matsui, Hirofumi; Kanamori, Toshiyuki

    2014-05-07

    Cell sorting is an essential and efficient experimental tool for the isolation and characterization of target cells. A three-dimensional environment is crucial in determining cell behavior and cell fate in biological analysis. Herein, we have applied photodegradable hydrogels to optical cell separation from a 3D environment using a computer-controlled light irradiation system. The hydrogel is composed of photocleavable tetra-arm polyethylene glycol and gelatin, which optimized cytocompatibility to adjust a composition of crosslinker and gelatin. Local light irradiation could degrade the hydrogel corresponding to the micropattern image designed on a laptop; minimum resolution of photodegradation was estimated at 20 µm. Light irradiation separated an encapsulated fluorescent microbead without any contamination of neighbor beads, even at multiple targets. Upon selective separation of target cells in the hydrogels, the separated cells have grown on another dish, resulting in pure culture. Cell encapsulation, light irradiation and degradation products exhibited negligible cytotoxicity in overall process.

  19. Mycotoxins of Aspergillus fumigatus in pure culture and in native bioaerosols from compost facilities.

    Science.gov (United States)

    Fischer, G; Müller, T; Ostrowski, R; Dott, W

    1999-04-01

    Exposure to secondary metabolites of airborne fungi in waste handling facilities is discussed in regard to potential toxic impacts on human health. The relevance of mycotoxins has been intensely studied in connection with contamination of food and feed. Toxic secondary metabolites are expected to be present in airborne spores, but exposure to mycotoxins in bioaerosols has not been studied sufficiently. Aspergillus fumigatus is one of the most frequent species in the air of compost plants. A wide range of secondary metabolites was found in pure cultures of freshly isolated strains of A. fumigatus. Tryptoquivaline, a compound with tremorgenic properties, and trypacidin, for which no toxic properties are described, were found in native bioaerosols in a compost facility. The highly toxic metabolites gliotoxin and verruculogen were not found in the bioaerosols.

  20. Effects of SHIKONIN on Cel Proliferation and Melanin Transfer of Co-Cultured Human Melanocytes and Keratinocytes%紫草素对黑素转运的影响

    Institute of Scientific and Technical Information of China (English)

    解士海; 潘武辉; 宗国勇; 黄小雄

    2014-01-01

    目的:探讨紫草素(Sh)在人表皮共培养体系中对黑素转运的影响。方法采用MTT法测定紫草素对人黑素细胞(MC)与角质形成细胞(KC)共培养体系增殖的影响,利用荧光标记技术以及流式细胞仪研究紫草素对于黑素转运的影响。结果0.25μmol/L、0.5μmol/L和1μmol/L紫草素对于人表皮共培养体系增殖无影响,0.25μmol/L、0.5μmol/L和1μmol/L紫草素对于共培养体系中黑素转运有剂量依赖地促进作用。结论紫草素对于共培养体系中黑素转运有剂量依赖的促进作用。%Objective: To study the Effects of SHIKONIN on cellProliferation and Melanin transfer of Co-Cultured of melanocytes and keratinocytes Methods: MTT method, Flow cytometry were employed to measure the cellviability, melanin transfer of co-cultured melanocytes and keratinocytes Results No influence was observed on co-cultured melanocytes and keratinocytes cellproliferation by 0.25μmol/L, 0.5μmol/L, or 1μmol/L of SHIKONIN . The melanin transfer were clearly enhanced by SHIKONIN in a dose-dependent manner at concentrations of 0.25μmol/L, 0.5μmol/L, and 1μmol/L Conclusion SHIKONIN exhibits a dose-dependent enhancing effects on melanin transfer in co-cultured melanocytes and keratinocytes.

  1. Fermentative hydrogen production from glucose and starch using pure strains and artificial co-cultures ofClostridium spp.

    Directory of Open Access Journals (Sweden)

    Masset Julien

    2012-05-01

    Full Text Available Abstract Background Pure bacterial strains give better yields when producing H2 than mixed, natural communities. However the main drawback with the pure cultures is the need to perform the fermentations under sterile conditions. Therefore, H2 production using artificial co-cultures, composed of well characterized strains, is one of the directions currently undertaken in the field of biohydrogen research. Results Four pure Clostridium cultures, including C. butyricum CWBI1009, C. pasteurianum DSM525, C. beijerinckii DSM1820 and C. felsineum DSM749, and three different co-cultures composed of (1 C. pasteurianum and C. felsineum, (2 C. butyricum and C. felsineum, (3 C. butyricum and C. pasteurianum, were grown in 20 L batch bioreactors. In the first part of the study a strategy composed of three-culture sequences was developed to determine the optimal pH for H2 production (sequence 1; and the H2-producing potential of each pure strain and co-culture, during glucose (sequence 2 and starch (sequence 3 fermentations at the optimal pH. The best H2 yields were obtained for starch fermentations, and the highest yield of 2.91 mol H2/ mol hexose was reported for C. butyricum. By contrast, the biogas production rates were higher for glucose fermentations and the highest value of 1.5 L biogas/ h was observed for the co-culture (1. In general co-cultures produced H2 at higher rates than the pure Clostridium cultures, without negatively affecting the H2 yields. Interestingly, all the Clostridium strains and co-cultures were shown to utilize lactate (present in a starch-containing medium, and C. beijerinckii was able to re-consume formate producing additional H2. In the second part of the study the co-culture (3 was used to produce H2 during 13 days of glucose fermentation in a sequencing batch reactor (SBR. In addition, the species dynamics, as monitored by qPCR (quantitative real-time PCR, showed a stable coexistence of C. pasteurianum and C

  2. Giant Congenital Melanocytic Nevus

    DEFF Research Database (Denmark)

    Rasmussen, Bo Sonnich; Henriksen, Trine Foged; Kølle, Stig-Frederik Trojahn

    2015-01-01

    Giant congenital melanocytic nevi (GCMN) occur in 1:20,000 livebirths and are associated with increased risk of malignant transformation. The treatment of GCMN from 1981 to 2010 in a tertiary referral center was reviewed evaluating the modalities used, cosmetic results, associated complications...

  3. 不含佛波酯和霍乱毒素条件下体外培养小儿正常表皮黑素细胞的实验研究%Children Normal Epidermal Melanocytes Culture Technology without Adding TPA and CT

    Institute of Scientific and Technical Information of China (English)

    李海翩; 陈谨萍; 罗佩谊; 张金桃; 李军

    2011-01-01

    [Objective] To research and explore the culture and amplification technology of children normal epidermal melanocytes without adding phorbol ester (TPA) and cholera toxin (CT),to identify the function of melanin synthesis and surface antigen production of melanocytes. [Methods] 15 cases of normal children skin tissues were obtained, from which melanocytes were separated and extracted, and the melanocytes were cultured and proliferated in M2 medium with adding basic fibroblast growth factor (bFGF), then DOPA staining and malanocytes (MART-1) fluorescent specific antibody binding assay were used to value melanin synthesis and the biological function of melanocytes surface antigen expressing in vitro. [Results] 15 healthy children autologous melanocytes were all successfully cultured,without the pollution of fibroblasts and keratinocytes. After proliferating 3-5 generations, the quantity of melanocytes meet the magnitude(80-100)thousand/cm2 that transplantation was required, and they could been continuously passaged in culture for about twenty generations. The staining result by DOPA of melanocytes were positive, uniform concentration, high purity, and melanocytes were able to combine with melanocyte specific MART-1 antibody. [Conclusion] Without adding TPA and CT in M2 medium the culture and proliferation of melanocytes made melanocytes activation growth status, vital cell quantity and biological functions very well, so these melanocytes could synthesize melanin and successfully express melanocyte surface antigen.%[目的]研究探索不含佛波酯、霍乱毒素条件下小儿正常表皮黑素细胞培养与扩增技术,鉴定黑素细胞合成黑素与产生表面抗原的功能,为开展小儿白癜风以及先天性表皮色素减退等疾病的细胞移植治疗提供可靠的实验依据。[方法]获取15例正常儿童的皮肤组织,分离提取黑素细胞,应用M2培养基并添加bFGF进行黑素细胞培养与增殖,采用DOPA染色法

  4. Isotopologue effects during N2O reduction in soils and in pure cultures of denitrifiers

    Science.gov (United States)

    Ostrom, Nathaniel E.; Pitt, Adam; Sutka, Robin; Ostrom, Peggy H.; Grandy, A. Stuart; Huizinga, Kristin M.; Robertson, G. Philip

    2007-06-01

    Site preference (SP), the difference in δ15N between the central and outer nitrogen atoms in N2O, is a powerful approach for apportioning fluxes of N2O from soils to nitrification and denitrification (Sutka et al., 2006). A critical aspect of the use of SP data to apportion sources of N2O to nitrification and denitrification is the need to evaluate data for isotope shifts that may have occurred during N2O reduction in soils prior to its escape to the atmosphere. We present data on the isotopologue effects during reduction of N2O during anaerobic incubation of soils and pure cultures of denitrifying bacteria. Isotopic enrichment factors for N2O reduction in soil mesocosms experiments varied between -9.2 and -1.8‰ for nitrogen and between -25.1 and -5.1‰ for oxygen. In pure cultures of Psuedomonas stutzeri and Psuedomonas denitrificans we observed isotopic enrichment factors for SP of -5.0 and -6.8‰, respectively. We further find that N2O consumption produces consistent relationships between δ18O and δ15N and δ18O and the δ15N of the central N atom in N2O of 2.5 and 1.6, respectively, which are clearly diagnostic of this process. Our results indicate that SP may be altered during reduction of N2O and thus bias evaluations of its origins. To understand the impacts of N2O reduction in soil flux studies on source isotope signals we modeled the isotope effects of N2O production occurring simultaneous with reduction and find increasingly curvilinear relationships between δ18O and δ15N and δ18O and δ15Nα with increased reduction. Consequently, a deviation from the linear mixing relationship between soil-derived and atmospheric N2O is an indication of extensive reduction. On the basis of our characterization of isotopic fractionation during N2O reduction, we show that the rate of reduction would have to be substantially greater than 10% of that of production to impact SP estimates of N2O from denitrification by more than a few percent. Nonetheless, reduction

  5. Fermentative hydrogen production by Clostridium butyricum CWBI1009 and Citrobacter freundii CWBI952 in pure and mixed cultures

    Directory of Open Access Journals (Sweden)

    Beckers, L.

    2010-01-01

    Full Text Available This paper investigates the biohydrogen production by two mesophilic strains, a strict anaerobe (Clostridium butyricum CWBI1009 and a facultative anaerobe (Citrobacter freundii CWBI952. They were cultured in pure and mixed cultures in serum bottles with five different carbon sources. The hydrogen yields of pure C. freundii cultures ranged from 0.09 molH2.molhexose-1 (with sucrose to 0.24 molH2.molhexose-1 (with glucose. Higher yields were obtained by the pure cultures of Cl. butyricum ranging from 0.44 molH2.molhexose-1 (with sucrose to 0.69 molH2.molhexose-1 (with lactose. This strain also fermented starch whereas C. freundii did not. However, it consumed the other substrates faster and produced hydrogen earlier than Cl. butyricum. This ability has been used to promote the growth conditions of Cl. butyricum in co-culture with C. freundii, since Cl. butyricum is extremely sensitive to the presence of oxygen which strongly inhibits H2 production. This approach could avoid the addition of any expensive reducing agents in the culture media such as L-cysteine since C. freundii consumes the residual oxygen. Thereafter, co-cultures with glucose and starch were investigated: hydrogen yields decreased from 0.53 molH2.molhexose-1 for pure Cl. butyricum cultures to 0.38 molH2.molhexose-1 for mixed culture with glucose but slightly increased with starch (respectively 0.69 and 0.73 molH2.molhexose-1. After 48 h of fermentation, metabolites analysis confirmed with microbial observation, revealed that the cell concentration of C. freundii dramatically decreased or was strongly inhibited by the development of Cl. butyricum.

  6. MITF accurately highlights epidermal melanocytes in atypical intraepidermal melanocytic proliferations.

    Science.gov (United States)

    Nybakken, Grant E; Sargen, Michael; Abraham, Ronnie; Zhang, Paul J; Ming, Michael; Xu, Xiaowei

    2013-02-01

    Atypical intraepidermal melanocytic proliferations (AIMP) have random cytologic atypia and other histologic features that are concerning for malignancy and often require immunohistochemistry to differentiate from melanoma in situ. Immunostaining with S100, Melan-A, and microphthalmia-associated transcription factor (MITF) was performed for 49 morphologically well-characterized AIMP lesions. The percentage of cells in the basal layer of the epidermis that were identified as melanocytes by immunohistochemistry was compared with the percentage observed by morphology on hematoxylin and eosin staining, which is the gold standard stain for identifying cytologic atypia within an AIMP. Melan-A estimated the highest percentage of melanocytes and S100 the fewest in 47 of the 49 lesions examined. The estimated percentage of melanocytes was 23.3% (95% confidence interval: 18.6-28.1; P Melanocyte estimates were similar for hematoxylin and eosin and MITF (P = 0.15) although S100 estimated 21.8% (95% confidence interval: -27.2 to -16.4; P melanocytes than hematoxylin and eosin. Melan-A staining produces higher estimates of epidermal melanocytes than S100 and MITF, which may increase the likelihood of diagnosing melanoma in situ. In contrast, melanoma in situ may be underdiagnosed with the use of S100, which results in lower estimates of melanocytes than the other 2 immunostains. Therefore, the best immunohistochemical marker for epidermal melanocytes is MITF.

  7. Pure culture of Metarhizium anisopliae LHL07 reprograms soybean to higher growth and mitigates salt stress.

    Science.gov (United States)

    Khan, Abdul Latif; Hamayun, Muhammad; Khan, Sumera Afzal; Kang, Sang-Mo; Shinwari, Zabta Khan; Kamran, Muhammad; Ur Rehman, Shafiq; Kim, Jong-Guk; Lee, In-Jung

    2012-04-01

    Little is known about the role of endophytic fungi against abiotic stresses and isoflavonoids (IF) contents of soybean. In current study, we investigated the role of fungal endophytes on the growth of soybean under salt stress conditions. Pure cultures of nine endophytic fungi were isolated from the roots of field-grown soybean plants, and their culture filtrates were screened on Waito-C and Dongjin-byeo rice cultivars; for identification of plant growth promoting fungal strains. It was observed that fungal isolate GMC-2B significantly promoted the growth of both Waito-C and Dongjin-byeo. GMC-2B was later identified as a new strain of Metarhizium anisopliae LHL07 on the basis of 18S rDNA sequences and phylogenetic analysis. Metarhizium anisopliae LHL07 inoculated soybean plants recorded significantly higher shoot length, shoot fresh and dry biomass, chlorophyll contents, transpiration rate, photosynthetic rate and leaf area; under sodium chloride induced salt stress as compared to non-inoculated control plants. An elevated proline and reduced superoxide dismutase and malondialdehyde contents in M. anisopliae LHL07 inoculated soybean plants demonstrated mitigation of salt induced oxidative stress. Furthermore, reduced abscisic acid and elevated jasmonic acid contents in soybean plants confirmed that lesser stress was convened to M. anisopliae inoculated-plants under salinity stress. We also assessed the role of M. anisopliae interaction on IF biosynthesis of soybean, and found significantly higher IF contents in M. anisopliae inoculated soybean plants. In conclusion, endophytic fungal interactions with soybean can be beneficial to improve soybean quality and quantity under salt affected agricultural systems.

  8. Mercury Stable Isotopic Composition of Monomethylmercury in Estuarine Sediments and Pure Cultures of Mercury Methylating Bacteria

    Science.gov (United States)

    Janssen, S.; Johnson, M. W.; Barkay, T.; Blum, J. D.; Reinfelder, J. R.

    2014-12-01

    Tracking monomethylmercury (MeHg) from its source in soils and sediments through various environmental compartments and transformations is critical to understanding its accumulation in aquatic and terrestrial food webs. Advances in the field of mercury (Hg) stable isotopes have allowed for the tracking of discrete Hg sources and the examination of photochemical and bacterial transformations. Despite analytical advances, measuring the Hg stable isotopic signature of MeHg in environmental samples or laboratory experiments remains challenging due to difficulties in the quantitative separation of MeHg from complex matrices with high concentrations of inorganic Hg. To address these challenges, we have developed a MeHg isolation method for sediments and bacterial cultures which involves separation by gas chromatography. The MeHg eluting from the GC is passed through a pyrolysis column and purged onto a gold amalgam trap which is then desorbed into a final oxidizing solution. A MeHg reference standard carried through our separation process retained its isotopic composition within 0.02 ‰ for δ202Hg, and for native estuarine sediments, MeHg recoveries were 80% to 100%. For sediment samples from the Hackensack and Passaic Rivers (New Jersey, USA), δ202Hg values for MeHg varied from -1.2 to +0.58 ‰ (relative to SRM 3133) and for individual samples were significantly different from that of total Hg (-0.38 ± 0.06 ‰). No mass independent fractionation was observed in MeHg or total Hg from these sediments. Pure cultures of Geobacter sulfurreducens, grown under fermentative conditions showed preferential enrichment of lighter isotopes (lower δ202Hg) during Hg methylation. The Hg stable isotope signatures of MeHg in sediments and laboratory methylation experiments will be discussed in the context of the formation and degradation of MeHg in the environment and the bioaccumulation of MeHg in estuarine food webs.

  9. Expression profiling of human melanocytes in response to UV-B irradiation

    Directory of Open Access Journals (Sweden)

    Saioa López

    2015-12-01

    Full Text Available A comprehensive gene expression analysis of human melanocytes was performed assessing the transcriptional profile of dark melanocytes (DM and light melanocytes (LM at basal conditions and after UV-B irradiation at different time points (6, 12 and 24 h, and in culture with different keratinocyte-conditioned media (KCM+ and KCM−. The data, previously published in [1], have been deposited in NCBI's Gene Expression Omnibus (GEO accession number: GSE70280.

  10. Degradation of the Ferric Chelate of EDTA by a Pure Culture of an Agrobacterium sp.

    Science.gov (United States)

    Lauff, J J; Steele, D B; Coogan, L A; Breitfeller, J M

    1990-11-01

    A pure culture of an Agrobacterium sp. (deposited as ATCC 55002) that mineralizes the ferric chelate of EDTA (ferric-EDTA) was isolated by selective enrichment from a treatment facility receiving industrial waste containing ferric-EDTA. The isolate grew on ferric-EDTA as the sole carbon source at concentrations exceeding 100 mM. As the degradation proceeded, carbon dioxide, ammonia, and an unidentified metabolite(s) were produced; the pH increased, and iron was precipitated from solution. The maximum rate of degradation observed with sodium ferric-EDTA as the substrate was 24 mM/day. At a substrate concentration of 35 mM, 90% of the substrate was degraded in 3 days and 70% of the associated chemical oxygen demand was removed from solution. Less than 15% of the carbon initially present was incorporated into the cell mass. Significant growth of this strain was not observed with uncomplexed EDTA as the sole carbon source at comparable concentrations; however, the ferric chelate of propylenediaminetetraacetic acid (ferric-PDTA) did support growth.

  11. Degradation of the ferric chelate of EDTA by a pure culture of an Agrobacterium sp

    Energy Technology Data Exchange (ETDEWEB)

    Lauff, J.J.; Coogan, L.A.; Breitfeller, J.M. (Genencor International, Rochester, NY (USA)); Steele, D.B. (Auburn Univ., AL (USA))

    1990-11-01

    A pure culture of an Agrobacterium sp. (deposited as ATCC 55002) the mineralizes the ferric chelate of EDTA (ferric-EDTA) was isolated by selective enrichment from a treatment facility receiving industrial waste containing ferric-EDTA. The isolated grew on ferric-EDTA as the sole carbon source at concentrations exceeding 100 mM. As the degradation proceeded, carbon dioxide, ammonia, and an unidentified metabolite(s) were produced; the pH increased, and iron was precipitated from solution. The maximum rate of degradation observed with sodium ferric-EDTA as the substrate was 24 mM/day. At a substrate concentration of 35 mM, 90% of the substrate was degraded in 3 days and 70% of the associated chemical oxygen demand was removed from solution. Less than 15% of the carbon initially present was incorporated into the cell mass. Significant growth of this strain was not observed with uncomplexed EDTA as the sole carbon source at comparable concentrations; however, the ferric chelate of propylenediaminetetraacetic acid (ferric-PDTA) did support growth.

  12. Skin melanocytes: biology and development

    OpenAIRE

    Cichorek, Mirosława; Wachulska, Małgorzata; Stasiewicz, Aneta; Tymińska, Agata

    2013-01-01

    In the human skin, melanocytes are present in the epidermis and hair follicles. The basic features of these cells are the ability to melanin production and the origin from neural crest cells. This last element is important because there are other cells able to produce melanin but of different embryonic origin (pigmented epithelium of retina, some neurons, adipocytes). The life cycle of melanocyte consists of several steps including differentiation of melanocyte lineage/s from neural crest, mi...

  13. Monitoring human melanocytic cell responses to piperine using multispectral imaging

    Science.gov (United States)

    Samatham, Ravikant; Phillips, Kevin G.; Sonka, Julia; Yelma, Aznegashe; Reddy, Neha; Vanka, Meenakshi; Thuillier, Philippe; Soumyanath, Amala; Jacques, Steven

    2011-03-01

    Vitiligo is a depigmentary disease characterized by melanocyte loss attributed most commonly to autoimmune mechanisms. Currently vitiligo has a high incidence (1% worldwide) but a poor set of treatment options. Piperine, a compound found in black pepper, is a potential treatment for the depigmentary skin disease vitiligo, due to its ability to stimulate mouse epidermal melanocyte proliferation in vitro and in vivo. The present study investigates the use of multispectral imaging and an image processing technique based on local contrast to quantify the stimulatory effects of piperine on human melanocyte proliferation in reconstructed epidermis. We demonstrate the ability of the imaging method to quantify increased pigmentation in response to piperine treatment. The quantization of melanocyte stimulation by the proposed imaging technique illustrates the potential use of this technology to quickly assess therapeutic responses of vitiligo tissue culture models to treatment non-invasively.

  14. Nanostructural change of human melanocytes stimulated with α-melanocyte stimulating hormone observed by atomic force microscopy.

    Science.gov (United States)

    Shin, Min Kyung; Lim, Hee Kyeong; Kang, Boo Kyoung; Lee, Gi-Ja; Uhm, Yun-Kyung; Lee, Mu-Hyoung

    2014-01-01

    It is well known that α-melanocyte stimulating hormone (α-MSH) induces melanization, but structural changes of melanocytes after α-MSH exposure are not well known. This study investigated the serial morphologic changes of human cultured melanocytes after stimulation with α-MSH using atomic force microscopy (AFM). Cultured human melanocytes were treated with α-MSH for 7 days. Contact mode AFM images were obtained using a NANOstation II and fixed melanocytes were scanned in PBS solution at a resolution of 512 × 512 pixels, at a scan speed of 0.4 line/s. The surface roughness and pore-like structures of the melanocytes were measured from topographic images using the Scanning Probe Imaging Processor. The surface roughness of cell body did not change significantly over time. All three roughness parameters (Sa, Sq, Sz) inside the pore-like structures on the dendrites increased significantly. The number of pore-like structures increased and the length and the breadth of pore-like structures also increased gradually. Pore-like structures changed statistically significantly in response to α-MSH stimulation. Further study will be needed to clarify whether the nature of pore-like structures is related to movement of melanosomes.

  15. Optimization of Differential Display of Prokaryotic mRNA: Application to Pure Culture and Soil Microcosms

    Science.gov (United States)

    Fleming, James T.; Yao, Wen-Hsiang; Sayler, Gary S.

    1998-01-01

    The differential display (DD) technique, which is widely used almost exclusively for eukaryotic gene discovery, was optimized to detect differential mRNA transcription from both pure-culture and soil-derived bacterial RNA. A model system which included toluene induction of todC1 in Pseudomonas putida F1 was used to optimize the procedure. At 24-h tod induction was determined to be approximately 8 × 107 transcripts/μg or 0.08% of the total mRNA. The primer concentration, primer length, annealing temperature, and template, deoxynucleoside triphosphate, and MgCl2 concentrations were varied to optimize amplification of a todC1 fragment. The limit of detection of todC1 by DD was found to be 0.015 ng of total RNA template or approximately 103 transcripts. Once optimized, a todC1C2 gene fragment from P. putida F1 RNA was detected by using an arbitrary primer for the reverse transcriptase step in conjunction with the same arbitrary primer and a Shine-Dalgarno primer in the PCR. To verify the results, an arbitrary primer was used to detect recovery of a new salicylate-inducible naphthalene dioxygenase in Burkholderia cepacia JS150. The method was then used to detect mRNA induction in both inoculated and uninoculated toluene-induced soil microcosms. Several putative differentially expressed partial gene sequences obtained from the uninoculated microcosms were examined, and one novel fragment was found to be differentially expressed. PMID:9758787

  16. Factors Affecting Volatile Phenol Production During Fermentations with Pure and Mixed Cultures of Dekkera bruxellensis and Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Janez Kosel

    2014-01-01

    Full Text Available The paper examines the impact of ethanol, and hidroxycinnamic and vinylphenol precursors on the production of volatile phenols in fermentations of mixed and pure cultures of yeasts Saccharomyces cerevisiae and Dekkera bruxellensis. Three different D. bruxellensis strains were examined and they all showed a unique volatile phenol production pattern in the fermentations of pure and mixed cultures. Generally, the results showed that in mixed culture fermentations less vinylphenols and more ethylphenols were produced in comparison with D. bruxellensis pure culture fermentations. Vinylphenol precursors significantly inhibited the growth of S. cerevisiae and the production of ethylphenols. Nevertheless, it was found that D. bruxellensis genes encoding for enzymes coumaric acid decarboxylase (CAD and vinylphenol reductase (VPR are more responsive to vinylphenol precursors in comparison with hidroxycinnamic acids. Consequently, higher concentrations of vinylphenols in the cell were found to be more cytotoxic than hidroxycinnamic acids. In general, low ethanol concentrations induced higher production of volatile phenols by S. cerevisiae and D. bruxellensis. This was confirmed with the expression pattern of gene encoding for CAD of D. bruxellensis.

  17. The efficacy of X-ray does on murine norovirus-1 (MNV-1) in pure culture, half-shell oyster, salmon sushi, and tuna salad

    Science.gov (United States)

    In this investigation, we determined the efficacy of X-ray doses on reducing a human norovirus (HuNoV) surrogate [murine norovirus-1 (MNV-1)] in pure culture, half-shell oyster, salmon sushi and tuna salad. The pure culture (phosphate-buffer saline, pH 7.4), half-shell oyster, salmon sushi and tuna ...

  18. Oxidation of acetate through reactions of the citric acid cycle by Geobacter sulfurreducens in pure culture and in syntrophic coculture

    OpenAIRE

    2000-01-01

    Geobacter sulfurreducens strain PCA oxidized acetate to CO2 via citric acid cycle reactions during growth with acetate plus fumarate in pure culture, and with acetate plus nitrate in coculture with Wolinella succinogenes. Acetate was activated by succinyl-CoA:acetate CoA-transferase and also via acetate kinase plus phosphotransacetylase. Citrate was formed by citrate synthase. Soluble isocitrate and malate dehydrogenases reduced NADP+ and NAD+, respectively. Oxidation of 2-oxoglutarate was me...

  19. Neocuproine Ablates Melanocytes in Adult Zebrafish

    OpenAIRE

    O'Reilly-Pol, Thomas; Johnson, Stephen L.

    2008-01-01

    The simplest regeneration experiments involve the ablation of a single cell type. While methods exist to ablate the melanocytes of the larval zebrafish,1,2 no convenient method exists to ablate melanocytes in adult zebrafish. Here, we show that the copper chelator neocuproine (NCP) causes fragmentation and disappearance of melanin in adult zebrafish melanocytes. Adult melanocytes expressing eGFP under the control of a melanocyte-specific promoter also lose eGFP fluorescence in the presence of...

  20. Prostaglandin induces the expression of matrix metalloproteinase-1 In ciliary melanocytes

    Institute of Scientific and Technical Information of China (English)

    WANG Ning-li; LU Qing-jun; LI Jun-hong; WANG Ling

    2008-01-01

    Background Latanoprost,a prostaglandin F2a analog,has been shown to be an effective intraocular pressure lowering agent which acts by inducing ciliary muscle cells to synthesise matrix metalloproteinases.However,the response of ciliary melanocytes to latanoprost has never been reported.This research has investigated the ability of latanoprost to induce matrix metalloproteinase-1 expression in human ciliary melanocytes,and thereby advance the understanding of the mechanism of PGF2a in decreasing Intraocular pressure.Methods In vitro human ciliary melanocytes were treated for 48 hours with five different concentrations of latanoprost (100,150,200,500,and 1000 nmol/L).Ciliary melanocytes treated with 0.01% ethanel(vehicle)were used as a control.The expression of matrix metalloproteinase-1 in ciliary melanocytes was determined by Western blotting and immunofluorescent staining.Results Western blotting showed that the expression of matrix metalloproteinase-1 in ciliary melanocytes was induced by latanoprost,and the level of expression was dependent on the concentration of latanoprost in the culture medium.Immunofluorescent staining showed that matrix metalloproteinase-1 was confined to the ciliary melanocyte cytoplasm.Conclusions Latanoprost induced the expression of matrix metalloproteinase-1 in human ciliary melanocytes in a dose-dependent manner.Ciliary melanocytes,as well as ciliary muscle cells,may also play an important role in uveoscleral outflow modulation.

  1. The role of the Hippo pathway in melanocytes and melanoma

    Directory of Open Access Journals (Sweden)

    Bruce Charles Baguley

    2013-05-01

    Full Text Available The Hippo signalling pathway comprises a series of cytoplasmic tumour suppressor proteins including Merlin and the Lats1/2 and MST1/2 kinases, and is thought to play a critical role in determining the sizes of organs and tissues. The Hippo pathway is regulated upstream by extracellular mechanosensory signalling arising from cell shape and polarity, as well as by a variety of extracellular signalling molecules. When active, the pathway maintains the transcriptional activators YAP and TAZ in phosphorylated forms in the cytoplasm, preventing cell proliferation. When the Hippo pathway is inactivated, YAP and TAZ are translocated to the nucleus and induce the expression of a variety of proteins concerned with entry into the cell division cycle, such as cyclin D1 and Fox M1, as well as the inhibition of apoptosis. The failure of the Hippo pathway has been implicated in the development of many different types of cancer but there is limited information available as to its involvement in melanoma. We hypothesise here firstly that the Hippo pathway is involved in maintaining density of cutaneous melanocytes on the basement membrane at the junction of the epidermis and the dermis, and secondly, that its function is disturbed in melanoma. We have analysed a series of 23 low passage human melanoma lines as well as in cultures of normal melanocytes, and find that melanocytes, as well as all melanoma cell lines examined express TAZ. Melanocytes and most melanoma lines also express YAP. E-cadherin, an upstream regulator of the Hippo pathway, and Axl, a receptor tyrosine kinase regulated by the Hippo pathway, are expressed in melanocytes and in several melanoma cell lines. These observations, together with published evidence for the presence of Merlin, Lats1/2 and MST1/2 in melanocytes and melanoma cells, support the hypothesis that the Hippo pathway is an important component of melanocyte and melanoma behaviour.

  2. [Effect of the fungicide captan on Azospirillum brasilense Cd in pure culture and associated with Setaria italica].

    Science.gov (United States)

    Di Ciocco, C A; Rodríguez Cáceres, E

    1997-01-01

    The effect of fungicide captan on growth and nitrogenase activity of Azospirillum brasilense Cd was studied in pure cultures and in association with foxtail millet (Setaria italica) cultivar Carapé under laboratory conditions. The 8 h growth in rotary shaker of A. brasilense was inhibited with 1 mg/l pure captan; however, after 4 days the differences diminished compared with the control without captan. Nitrogenase activity was affected with 10 mg/l but the differences were negligible after 48 h of growth. Root dry weight of inoculated plants was diminished by the treatment of foxtail millet cv. Carapé with captan. Inoculation with A. brasilense Cd increased shoot dry weight, but differences were significant only with respect to the control but not in relation to captan treatments.

  3. Effect of streptomycin on melanogenesis and antioxidant status in melanocytes.

    Science.gov (United States)

    Wrześniok, Dorota; Beberok, Artur; Otręba, Michał; Buszman, Ewa

    2013-11-01

    Streptomycin is an aminoglycoside antibiotic with an antituberculosis activity commonly used in clinical practice due to its good antimicrobial characteristics. A well-known undesirable side effect of this drug is ototoxicity, which may be caused by overproduction of reactive oxygen species and loss of melanocytes in the inner ear. The aim of this study was to examine the effect of streptomycin on melanogenesis and antioxidant defense system in cultured normal human melanocytes (HEMa-LP). Streptomycin induced concentration-dependent loss in melanocytes viability. The value of EC50 was determined to be ~5.0 mM. It has been shown that streptomycin causes inhibition of tyrosinase activity and reduces melanin content in human melanocytes in a concentration-dependent manner. Significant changes in the activity of cellular antioxidant enzymes: superoxide dismutase, catalase, and glutathione peroxidase were also stated. The results obtained in vitro may explain a potential role of melanocytes and melanin in the causative mechanisms of aminoglycosides ototoxic effects in vivo.

  4. Hesperetin induces melanin production in adult human epidermal melanocytes.

    Science.gov (United States)

    Usach, Iris; Taléns-Visconti, Raquel; Magraner-Pardo, Lorena; Peris, José-Esteban

    2015-06-01

    One of the major sources of flavonoids for humans are citrus fruits, hesperidin being the predominant flavonoid. Hesperetin (HSP), the aglycon of hesperidin, has been reported to provide health benefits such as antioxidant, anti-inflammatory and anticarcinogenic effects. However, the effect of HSP on skin pigmentation is not clear. Some authors have found that HSP induces melanogenesis in murine B16-F10 melanoma cells, which, if extrapolated to in vivo conditions, might protect skin against photodamage. Since the effect of HSP on normal melanocytes could be different to that observed on melanoma cells, the described effect of HSP on murine melanoma cells has been compared to the effect obtained using normal human melanocytes. HSP concentrations of 25 and 50 µM induced melanin synthesis and tyrosinase activity in human melanocytes in a concentration-dependent manner. Compared to control melanocytes, 25 µM HSP increased melanin production and tyrosinase activity 1.4-fold (p melanocyte cultures could be reproduced on human skin.

  5. 白介素-1对培养人表皮黑素细胞增殖及黑素合成的影响研究%Study on effects of Interleutin-1 on cell proliferation and melanin synthesis in cultured human melanocytes

    Institute of Scientific and Technical Information of China (English)

    林新瑜; 王芳; 林鸿刚; 段西凌

    2012-01-01

    Objective Establishing the cultured system of human normal melanocytes in vitro, and studying effects of IL-1a and IL-1p on cultured human melanocytes. Methods MTT method,NaOH assay were employed to measure the rate of proliferation, melanin synthesis of melanocytes respectively; apoptosis rates of melanocytes were detected by using flow cytometer. Results Not only IL-1a but also IL-1β showed a significant decrease of the rate of melanocyte proliferation in concentration and time- dependent manner, inhibited melanin synthesis and increased melanocyte apoptosis. Conclusion It suggests that not only IL-1o but also IL-1β at the concentration of 50-200ng/ml is able to suppress melanocytic proliferation and melanin synthesis. It may be developed to be a potential skin lightening agent and worth to study later.%目的:建立正常人表皮黑素细胞体外培养体系,观察不同浓度的白介素-1α,白介素-1 β对体外培养正常人黑素细胞的细胞增殖及黑素合成的影响.方法:采用四甲基偶氮唑盐(MTT)法测定白介素-1α及白介素-1 β对黑素细胞增值的影响,NaOH裂解法测定黑素生成量,流式细胞仪检测黑素细胞的凋亡率.结果:白介素-1α及白介素-1 p对黑素细胞活力有抑制作用,能使细胞增殖能力降低,黑素合成减少,增加黑素细胞的凋亡率.结论:白介素-1α及白介素-1β对体外培养的黑素细胞增殖及黑素生成都有抑制作用,这为临床应用白介素-1α及白介素-1β治疗色素性疾病提供了实验依据.

  6. Nicotinamide enhances repair of ultraviolet radiation-induced DNA damage in primary melanocytes.

    Science.gov (United States)

    Thompson, Benjamin C; Surjana, Devita; Halliday, Gary M; Damian, Diona L

    2014-07-01

    Cutaneous melanoma is a significant cause of morbidity and mortality. Nicotinamide is a safe, widely available vitamin that reduces the immune suppressive effects of UV, enhances DNA repair in keratinocytes and has shown promise in the chemoprevention of non-melanoma skin cancer. Here, we report the effect of nicotinamide on DNA damage and repair in primary human melanocytes. Nicotinamide significantly enhanced the repair of oxidative DNA damage (8-oxo-7,8-dihydro-2'-deoxyguanosine) and cyclobutane pyrimidine dimers induced by UV exposure. It also enhanced the repair of 8-oxo-7,8-dihydro-2'-deoxyguanosine induced by the culture conditions in unirradiated melanocytes. A significant increase in the percentage of melanocytes undergoing unscheduled but not scheduled DNA synthesis was observed, confirming that nicotinamide enhances DNA repair in human melanocytes. In summary, nicotinamide, by enhancing DNA repair in melanocytes, is a potential agent for the chemoprevention of cutaneous melanoma.

  7. Effects of imperatorin on Rab27a and tyrosinase in cultured human epider mal melanocytes%欧前胡素对体外培养的人表皮黑素细胞Rab27a和酪氨酸酶的作用

    Institute of Scientific and Technical Information of China (English)

    谢佳均; 佘绍逸; 林玲; 刘少刚; 吴晓佳; 阮海东; 郭佩湘; 童鹏飞; 方斌; 陈玮莹

    2014-01-01

    目的:观察欧前胡素对体外培养的人表皮黑素细胞转运相关蛋白Rab27a和酪氨酸酶的作用。方法:体外分离、纯化培养人表皮黑素细胞。随机分为空白对照组和欧前胡素处理组,以50或25μmol/L欧前胡素作用48 h后观察黑素细胞形态,免疫荧光法检测Rab27a和酪氨酸酶在黑素细胞中的定位及变化,应用透射电镜观察黑素细胞黑素小体的超微结构。结果:欧前胡素作用48 h后,黑素细胞的树状突起缩短,免疫荧光检测显示,黑素细胞Rab27a和酪氨酸酶蛋白在细胞内定位未见明显变化,但欧前胡素作用后Rab27a蛋白的荧光强度轻度增强(P<0.05),而酪氨酸酶荧光强度降低(<0.05)。透射电镜结果可见黑素细胞中的黑素小体数量减少。结论:欧前胡素对体外培养的人表皮黑素细胞Rab27a表达呈促进作用,对酪氨酸酶活性及黑素合成呈抑制作用。%To investigate the effects of imperatorin on Rab27a and tyrosinase in cultured human epidermal melanocytes. METHODS:Human epidermal melanocytes were separated,purified and cultured in vitro. Melanocytes were randomly divided into control group and imperatorin group. After 48 h incubation with culture medium M254 or imperatorin(50 or 25 µmol/L),the morphology of melanocytes was examined by inverted phase contrast microscopy. The effects of imperatorin on Rab27a and tyrosinase in melanocytes were investigated by immunofluoresent assay. The ultrastructures of melanocytes and melanosomes were studied by transmission electron microscopy. RESULTS:The melanocytes cultured with imperatorin for 48 h had shorter dendrites. Immunofluorescence microscopy showed that the location of Rab27a and tyrosinase in melanocytes were not obviously changed,but the fluorescence intensity of Rab27a was enhanced (P<0.05) and tyrosinase attenuated (P<0.05). Fewer melanosomes found in the cultured human epidermal melanocytes by transmission electron

  8. Genetics Home Reference: giant congenital melanocytic nevus

    Science.gov (United States)

    ... that is composed of pigment-producing cells called melanocytes . It is present from birth (congenital) or is ... is the presence of pigment-producing skin cells (melanocytes) in the tissue that covers the brain and ...

  9. Functional melanocytes are readily reprogrammable from multilineage-differentiating stress-enduring (muse) cells, distinct stem cells in human fibroblasts.

    Science.gov (United States)

    Tsuchiyama, Kenichiro; Wakao, Shohei; Kuroda, Yasumasa; Ogura, Fumitaka; Nojima, Makoto; Sawaya, Natsue; Yamasaki, Kenshi; Aiba, Setsuya; Dezawa, Mari

    2013-10-01

    The induction of melanocytes from easily accessible stem cells has attracted attention for the treatment of melanocyte dysfunctions. We found that multilineage-differentiating stress-enduring (Muse) cells, a distinct stem cell type among human dermal fibroblasts, can be readily reprogrammed into functional melanocytes, whereas the remainder of the fibroblasts do not contribute to melanocyte differentiation. Muse cells can be isolated as cells positive for stage-specific embryonic antigen-3, a marker for undifferentiated human embryonic stem cells, and differentiate into cells representative of all three germ layers from a single cell, while also being nontumorigenic. The use of certain combinations of factors induces Muse cells to express melanocyte markers such as tyrosinase and microphthalmia-associated transcription factor and to show positivity for the 3,4-dihydroxy-L-phenylalanine reaction. When Muse cell-derived melanocytes were incorporated into three-dimensional (3D) cultured skin models, they localized themselves in the basal layer of the epidermis and produced melanin in the same manner as authentic melanocytes. They also maintained their melanin production even after the 3D cultured skin was transplanted to immunodeficient mice. This technique may be applicable to the efficient production of melanocytes from accessible human fibroblasts by using Muse cells, thereby contributing to autologous transplantation for melanocyte dysfunctions, such as vitiligo.

  10. Fermentative capabilities and volatile compounds produced by Kloeckera/Hanseniaspora and Saccharomyces yeast strains in pure and mixed cultures during Agave tequilana juice fermentation.

    Science.gov (United States)

    González-Robles, Ivonne Wendolyne; Estarrón-Espinosa, Mirna; Díaz-Montaño, Dulce María

    2015-09-01

    The fermentative and aromatic capabilities of Kloeckera africana/Hanseniaspora vineae K1, K. apiculata/H. uvarum K2, and Saccharomyces cerevisiae S1 and S2 were studied in pure and mixed culture fermentations using Agave tequila juice as the culture medium. In pure and mixed cultures, Kloeckera/Hanseniaspora strains showed limited growth and sugar consumption, as well as low ethanol yield and productivity, compared to S. cerevisiae, which yielded more biomass, ethanol and viable cell concentrations. In pure and mixed cultures, S. cerevisiae presented a similar behaviour reaching high biomass production, completely consuming the sugar, leading to high ethanol production. Furthermore, the presence of S. cerevisiae strains in the mixed cultures promoted the production of higher alcohols, acetaldehyde and ethyl esters, whereas Kloeckera/Hanseniaspora strains stimulated the production of ethyl acetate and 2-phenyl ethyl acetate compounds.

  11. Simple screening method for molds producing intracellular mycotoxins in pure cultures.

    Science.gov (United States)

    Filtenborg, O; Frisvad, J C; Svendsen, J A

    1983-02-01

    A simple screening method for molds producing the intracellular mycotoxins brevianamide A, citreoviridin, cyclopiazonic acid, luteoskyrin, penitrem A, roquefortine C, sterigmatocystin, verruculogen, viomellein, and xanthomegnin was developed. After removing an agar plug from the mold culture, the mycelium on the plug is wetted with a drop of methanol-chloroform (1:2). By this treatment the intracellular mycotoxins are extracted within seconds and transferred directly to a thin-layer chromatography plate by immediately placing the plug on the plate while the mycelium is still wet. After removal of the plug, known thin-layer chromatographic procedures are carried out. The substrate (Czapek yeast autolysate agar) and growth conditions (25 degrees C for 7 days) used by Penicillium taxonomists proved suitable for the production of the mycotoxins investigated when 60 known toxigenic isolates and 865 cultures isolated from foods and feedstuffs were tested with this screening method.

  12. Human melanocytes mitigate keratinocyte-dependent contraction in an in vitro collagen contraction assay.

    Science.gov (United States)

    Rakar, Jonathan; Krammer, Markus P; Kratz, Gunnar

    2015-08-01

    Scarring is an extensive problem in burn care, and treatment can be especially complicated in cases of hypertrophic scarring. Contraction is an important factor in scarring but the contribution of different cell types remains unclear. We have investigated the contractile behavior of keratinocytes, melanocytes and fibroblasts by using an in vitro collagen gel assay aimed at identifying a modulating role of melanocytes in keratinocyte-mediated contraction. Cells were seeded on a collagen type I gel substrate and the change in gel dimensions were measured over time. Hematoxylin & Eosin-staining and immunohistochemistry against pan-cytokeratin and microphthalmia-associated transcription factor showed that melanocytes integrated between keratinocytes and remained there throughout the experiments. Keratinocyte- and fibroblast-seeded gels contracted significantly over time, whereas melanocyte-seeded gels did not. Co-culture assays showed that melanocytes mitigate the keratinocyte-dependent contraction (significantly slower and 18-32% less). Fibroblasts augmented the contraction in most assays (approximately 6% more). Non-contact co-cultures showed some influence on the keratinocyte-dependent contraction. Results show that mechanisms attributable to melanocytes, but not fibroblasts, can mitigate keratinocyte contractile behavior. Contact-dependent mechanisms are stronger modulators than non-contact dependent mechanisms, but both modes carry significance to the contraction modulation of keratinocytes. Further investigations are required to determine the mechanisms involved and to determine the utility of melanocytes beyond hypopigmentation in improved clinical regimes of burn wounds and wound healing.

  13. Extraction and characterization of bound extracellular polymeric substances from cultured pure cyanobacterium (Microcystis wesenbergii).

    Science.gov (United States)

    Liu, Lizhen; Qin, Boqiang; Zhang, Yunlin; Zhu, Guangwei; Gao, Guang; Huang, Qi; Yao, Xin

    2014-08-01

    Preliminary characterization of bound extracellular polymeric substances (bEPS) of cyanobacteria is crucial to obtain a better understanding of the formation mechanism of cyanobacterial bloom. However, the characterization of bEPS can be affected by extraction methods. Five sets (including the control) of bEPS from Microcystis extracted by different methods were characterized using three-dimensional excitation and emission matrix (3DEEM) fluorescence spectroscopy combined chemical spectrophotometry; and the characterization results of bEPS samples were further compared. The agents used for extraction were NaOH, pure water and phosphate buffered saline (PBS) containing cationic exchange resins, and hot water. Extraction methods affected the fluorescence signals and intensities in the bEPS. Five fluorescence peaks were observed in the excitation and emission matrix fluorescence spectra of bEPS samples. Two peaks (peaks T₁ and T₂) present in all extractions were identified as protein-like fluorophores, two (peaks A and C) as humic-like fluorophores, and one (peak E) as a fulvic-like substance. Among these substances, the humic-like and fulvic-like fluorescences were only seen in the bEPS extracted with hot water. Also, NaOH solution extraction could result in strong fluorescence intensities compared to the other extraction methods. It was suggested that NaOH at pH10.0 was the most appropriate method to extract bEPS from Microcystis. In addition, dialysis could affect the yields and characteristics of extracted bEPS during the determination process. These results will help us to explore the issues of cyanobacterial blooms.

  14. Classification of melanocytic skin lesions from non-melanocytic lesions.

    Science.gov (United States)

    Iyatomi, Hitoshi; Norton, Kerri-Ann; Celebi, M; Schaefer, Gerald; Tanaka, Masaru; Ogawa, Koichi

    2010-01-01

    In this paper, we present a classification method of dermoscopy images between melanocytic skin lesions (MSLs) and non-melanocytic skin lesions (NoMSLs). The motivation of this research is to develop a pre-processor of an automated melanoma screening system. Since NoMSLs have a wide variety of shapes and their border is often ambiguous, we developed a new tumor area extraction algorithm to account for these difficulties. We confirmed that this algorithm is capable of handling different dermoscopy images not only those of NoMSLs but also MSLs as well. We determined the tumor area from the image using this new algorithm, calculated a total 428 features from each image, and built a linear classifier. We found only two image features, "the skewness of bright region in the tumor along its major axis" and "the difference between the average intensity in the peripheral part of the tumor and that in the normal skin area using the blue channel" were very efficient at classifying NoMSLs and MSLs. The detection accuracy of MSLs by our classifier using only the above mentioned image feature has a sensitivity of 98.0% and a specificity of 86.6% in a set of 107 non-melanocytic and 548 melanocytic dermoscopy images using a cross-validation test.

  15. S100B as a potential biomarker for the detection of cytotoxicity of melanocytes.

    Science.gov (United States)

    Cheong, Kyung Ah; Noh, Minsoo; Kim, Chang-Hyun; Lee, Ai-Young

    2014-03-01

    Skin irritation is one of the most common adverse reactions in hydroquinone (HQ) and retinoic acid (RA). Although melanocytes have rarely been considered to be involved in skin irritation, RA and particularly HQ could induce melanocyte toxicity, resulting in depigmentation. We chose S100B as a candidate gene for melanocytotoxicity from a genome-wide transcriptional profiling analysis after applying irritant doses of HQ, RA and sodium lauryl sulphate (SLS) to cultures of keratinocytes and/or melanocytes. In this study, the role of S100B on melanocyte viability and cytotoxicity was examined. S100B was detected in melanocytes, but not in keratinocytes or fibroblasts. Melanocytes after treatment with increasing concentrations of HQ, RA, SLS and urushiol showed significant increases in intracellular and extracellular S100B expression with reduced viable cell number and increased release of lactate dehydrogenase. No RAGE expression and no significant function of CD166/ALCAM in melanocyte survival and cytotoxicity favoured the role of intracellular S100B in chemically irritated melanocytes. S100B knock-down increased apoptosis through inhibition of PI3K/AKT, NF-κB and ERK activation, suggesting the increased intracellular S100B expression by chemical irritation as a compensatory reaction to reduce cytotoxicity. The numerical decrease in S100B/c-kit-double-positive melanocytes was also examined in human skin epidermis irritated by HQ or RA with stronger staining intensities of S100B. Collectively, the decrease in viable cell number by reduced intracellular S100B levels in vitro and by chemical irritation in vivo suggests that S100B could be a potential biomarker for melanocytes cytotoxicity.

  16. Effects of Interleukin-4 on Cell Proliferation and Melanin Synthesis in Cultured Human Melanocytes in Vitro%白细胞介素4对体外培养人黑素细胞增殖及黑素合成的影响

    Institute of Scientific and Technical Information of China (English)

    陈静宇; 林新瑜; 王芳; 林鸿刚

    2012-01-01

    Objective To establish the cultured system of human normal melanocytes in vitro, and to study effects of interleukin-4(IL-4) on cultured human melanocytes. Methods Methyl thiazolyl tetrazolium(MTT) method, NaOH cracking process assay was employed to measure the rate of proliferation, melanin synthesis of melanocytes respectively; apoptosis rates of melanocytes were detected by flow cytometry. Results IL-4 showed a significant decreasing in the rate of melanocyte proliferation in concentration and time -dependent manner. It could inhibite melanin synthesis and increase melanocyte apoptosis. Conclusion It suggests that IL-4 at the concentration of 50 to 200 μg/L is able to suppress melanoeytic proliferation and melanin synthesis. This provides experiment reference for IL-4 on treating pigmentary diseases.%目的 建立正常人表皮黑素细胞体外培养体系,观察不同浓度白细胞介素4(interleukin-4,IL-4)对体外培养正常人黑素细胞的细胞增殖及黑素合成的影响.方法 采用四甲基偶氮唑盐比色(methylthiazolyl tetrazolium,MTT)法测定不同浓度IL-4对黑素细胞增殖的影响,氢氧压钠(NaOH)裂解法测定黑素生成量,流式细胞仪检测黑素细胞的凋亡率.结果 IL-4对黑素细胞活力有抑制作用,能使细胞增殖能力降低,黑素合成减少,增加黑素细胞的凋亡率.结论 IL-4对体外培养的黑素细胞增殖及黑素生成都有抑制作用,这为临床应用IL-4治疗色素性疾病提供了实验依据.

  17. Protein improvement in Gari by the use of pure cultures of microorganisms involved in the natural fermentation process.

    Science.gov (United States)

    Ahaotu, I; Ogueke, C C; Owuamanam, C I; Ahaotu, N N; Nwosu, J N

    2011-10-15

    The ability of microorganisms involved in cassava mash fermentation to produce and improve protein value by these microorganisms during fermentation was studied. Standard microbiological procedures were used to isolate, identify and determine the numbers of the organisms. Alcaligenes faecalis, Lactobacillus plantarum, Bacillus subtilis, Leuconostoc cremoris, Aspergillus niger, A. tamari, Geotrichum candidum and Penicillium expansum were isolated and identified from cassava waste water while standard analytical methods were used to determine the ability of the isolates to produce linamarase and the proximate composition, pH and titrable acidity of the fermenting mash. The linamarase activity of the isolates ranged from 0.0416 to 0.2618 micromol mL(-1) nmol(-1). Bacillus subtilis, A. niger, A. tamari and P. expansum did not express any activity for the enzyme. Protein content of mash fermented with mixed fungal culture had the highest protein value (15.4 mg/g/dry matter) while the raw cassava had the least value (2.37 mg/g/dry matter). The naturally fermented sample had the least value for the fermented samples (3.2 mg/g/dry matter). Carbohydrate and fat contents of naturally fermented sample were higher than values obtained from the other fermented samples. Microbial numbers of the sample fermented with mixed bacterial culture was highest and got to their peak at 48 h (57 x 10(8) cfu g(-1)). pH decreased with increase in fermentation time with the mash fermented by the mixed culture of fungi having the lowest pH of 4.05 at the end of fermentation. Titrable acidity increased with increase in fermentation time with the highest value of 1.32% at 96 h of fermentation produced by the mixed culture of fungi. Thus fermentation with the pure cultures significantly increased the protein content of mash.

  18. Degradation and mineralization of wheat straw by some cellulolytic fungi in pure cultures

    Energy Technology Data Exchange (ETDEWEB)

    Omar, S.A. (Botany Dept., Faculty of Science, Assiut Univ. (Egypt))

    1994-01-01

    Decomposition and mineralization of wheat straw inoculated with 8 cellulolytic fungi was investigated in sand culture. Aspergillus fumigatus and Stachybotrys chartarum showed a great potential to degrade wheat straw and weight loss of straw was 35.3 and 27.5% of initial weight after 63 days incubation, respectively. CO[sub 2] production was highest after 7 and 14 days for all tested fungi suggesting that the main contribution to CO[sub 2] production arose from the water-soluble fraction which was decomposed rapidly. Carbon solubilization from wheat straw occurred, to some extent, in a manner similar to that of straw decomposition. Also, S. chartarum and A. fumigatus solubilized considerable amounts of straw Nitrogen but Gibberella fujikuroi was the best nitrogen dissolving organism. (orig.)

  19. Anaerobic degradation of o-phenylphenol by mixed and pure cultures

    Energy Technology Data Exchange (ETDEWEB)

    Sembiring, T.; Winter, J.

    1989-07-01

    An anaerobic enrichment culture that degrade 0.4 mmol/l per day of o-phenylphenol was selected from sediment of a waste water pond of a sugar factory. From the consortium an o-phenylphenol-degrading bacterium, strain B10, was isolated. Strain B10 could not degrade other aromatic substances, including phenylacetic acid, benzoate, o-hydroxybenzoate, p-hydroxybenzoate and phenol. Best growth was observed with glucose, pyruvate, lactate, methanol and H/sub 2//CO/sub 2/ as substrates, o-Phenylphenol was slowly degraded if supplied as the only carbon source and was cometabolized in the presence of >5 mmol/l glucose. Strain B10 has not yet been assigned to a known species or family.

  20. Biodegradation of Synthetic Polyesters (BTA and PCL with Natural Flora in Soil Burial and Pure Cultures under Ambient Temperature

    Directory of Open Access Journals (Sweden)

    Mona K. Gouda

    2012-03-01

    Full Text Available The aim of this study was to study the biodegradation of two synthetic polyesters, one aliphaticaromatic (1, 4-butanediol, terephthalic-adipic acid, BTA and the other aliphatic (poly (,-caprolactone, PCL, under different soil types (canal shore soil, garden soil, compost and Peat moss, respectively, as well as using locally isolated cultures at ambient temperature. The results showed that the BTA films buried in canal shore and garden soil were degraded faster than that in the other soils. After six weeks about 90, 88 and 80% were degraded in garden, canal shore soil and compost respectively, while only 52% were degraded in Peat moss. On the other hand, 95 and 93% weight loss was obtained for PCL films buried for three weeks in canal shore and garden soil respectively. The Scanning Electron Microscope photos confirm the results of weight loss and revealed the presence of cracks and fungal growth on films buried in different soils. The results with pure cultures, especially with Fusarium solani, also confirmed the biodegradability of two polyesters under ambient temperature. Finally, it could be concluded that both synthetic polyester are degradable under ambient conditions.

  1. Melanocytes Affect Nodal Expression and Signaling in Melanoma Cells: A Lesson from Pediatric Large Congenital Melanocytic Nevi.

    Science.gov (United States)

    Margaryan, Naira V; Gilgur, Alina; Seftor, Elisabeth A; Purnell, Chad; Arva, Nicoleta C; Gosain, Arun K; Hendrix, Mary J C; Strizzi, Luigi

    2016-03-22

    Expression of Nodal, a Transforming Growth Factor-beta (TGF-β) related growth factor, is associated with aggressive melanoma. Nodal expression in adult dysplastic nevi may predict the development of aggressive melanoma in some patients. A subset of pediatric patients diagnosed with giant or large congenital melanocytic nevi (LCMN) has shown increased risk for development of melanoma. Here, we investigate whether Nodal expression can help identify the rare cases of LCMN that develop melanoma and shed light on why the majority of these patients do not. Immunohistochemistry (IHC) staining results show varying degree of Nodal expression in pediatric dysplastic nevi and LCMN. Moreover, median scores from Nodal IHC expression analysis were not significantly different between these two groups. Additionally, none of the LCMN patients in this study developed melanoma, regardless of Nodal IHC levels. Co-culture experiments revealed reduced tumor growth and lower levels of Nodal and its signaling molecules P-SMAD2 and P-ERK1/2 when melanoma cells were grown in vivo or in vitro with normal melanocytes. The same was observed in melanoma cells cultured with melanocyte conditioned media containing pigmented melanocyte derived melanosomes (MDM). Since MDM contain molecules capable of inactivating radical oxygen species, to investigate potential anti-oxidant effect of MDM on Nodal expression and signaling in melanoma, melanoma cells were treated with either N-acetyl-l-cysteine (NAC), a component of the anti-oxidant glutathione or synthetic melanin, which in addition to providing pigmentation can also exert free radical scavenging activity. Melanoma cells treated with NAC or synthetic melanin showed reduced levels of Nodal, P-SMAD2 and P-ERK1/2 compared to untreated melanoma cells. Thus, the potential role for Nodal in melanoma development in LCMN is less evident than in adult dysplastic nevi possibly due to melanocyte cross-talk in LCMN capable of offsetting or delaying the pro

  2. A Study of Mercury Methylation Genetics: Qualitative and Quantitative Analysis of hgcAB in Pure Culture

    Science.gov (United States)

    Christensen, G. A.; Wymore, A. M.; King, A. J.; Podar, M.; Hurt, R. A., Jr.; Santillan, E. F. U.; Gilmour, C. C.; Brandt, C. C.; Brown, S. D.; Palumbo, A. V.; Elias, D. A.

    2015-12-01

    Two proteins (HgcA and HgcB) have been determined to be essential for mercury (Hg)-methylation and either one alone is not sufficient for this process. Detection and quantification of these genes to determine at risk environments is critical. Universal degenerate polymerase chain reaction (PCR) primers spanning hgcAB were developed to ascertain organismal diversity and validate that both genes were present as an established prerequisite for Hg-methylation. To confirm this approach, an extensive set of pure cultures with published genomes (including methylators and non-methylators: 13 Deltaproteobacteria, 9 Firmicutes, and 10 methanogenic Archaea) were assayed with the newly designed universal hgcAB primer set. A single band within an agarose gel was observed for the majority of the cultures with known hgcAB and confirmed via Sanger sequencing. For environmental applications, once the potential for Hg-methylation is established from PCR amplification with the universal hgcAB primer set, quantification of clade-specific hgcAB gene abundance is desirable. We developed quantitative polymerase chain reaction (qPCR) degenerate primers targeting hgcA from each of the three dominate clades (Deltaproteobacteria, Firmicutes and methanogenic Archaea) known to be associated with anaerobic Hg-methylation. The qPCR primers amplify virtually all hgcA positive cultures overall and are specific for their designed clade. Finally, to ensure the procedure is robust and sensitive in complex environmental matrices, cells from all clades were mixed in different combinations and ratios to assess qPCR primer specificity. The development and validation of these high fidelity quantitative molecular tools now allows for rapid and accurate risk management assessment in any environment.

  3. Microphthalmia-associated transcription factor as the molecular target of cadmium toxicity in human melanocytes

    Energy Technology Data Exchange (ETDEWEB)

    Chantarawong, Wipa [Department of Molecular Biology and Applied Physiology, Tohoku University School of Medicine, Sendai (Japan); Inter Departmental Multidisciplinary Graduate Program in Bioscience, Faculty of Science, Kasetsart University, Bangkok (Thailand); Takeda, Kazuhisa; Sangartit, Weerapon; Yoshizawa, Miki [Department of Molecular Biology and Applied Physiology, Tohoku University School of Medicine, Sendai (Japan); Pradermwong, Kantimanee [Department of Zoology, Faculty of Science, Kasetsart University, Bangkok (Thailand); Shibahara, Shigeki, E-mail: shibahar@med.tohoku.ac.jp [Department of Molecular Biology and Applied Physiology, Tohoku University School of Medicine, Sendai (Japan)

    2014-11-28

    Highlights: • In human melanocytes, cadmium decreases the expression of MITF-M and tyrosinase and their mRNAs. • In human melanoma cells, cadmium decreases the expression of MITF-M protein and tyrosinase mRNA. • Expression of MITF-H is less sensitive to cadmium toxicity in melanocyte-linage cells. • Cadmium does not decrease the expression of MITF-H in retinal pigment epithelial cells. • MITF-M is the molecular target of cadmium toxicity in melanocytes. - Abstract: Dietary intake of cadmium is inevitable, causing age-related increase in cadmium accumulation in many organs, including hair, choroid and retinal pigment epithelium (RPE). Cadmium has been implicated in the pathogenesis of hearing loss and macular degeneration. The functions of cochlea and retina are maintained by melanocytes and RPE, respectively, and the differentiation of these pigment cells is regulated by microphthalmia-associated transcription factor (MITF). In the present study, we explored the potential toxicity of cadmium in the cochlea and retina by using cultured human melanocytes and human RPE cell lines. MITF consists of multiple isoforms, including melanocyte-specific MITF-M and widely expressed MITF-H. Levels of MITF-M protein and its mRNA in human epidermal melanocytes and HMV-II melanoma cells were decreased significantly by cadmium. In parallel with the MITF reduction, mRNA levels of tyrosinase, the key enzyme of melanin biosynthesis that is regulated by MITF-M, were also decreased. In RPE cells, however, the levels of total MITF protein, constituting mainly MITF-H, were not decreased by cadmium. We thus identify MITF-M as the molecular target of cadmium toxicity in melanocytes, thereby accounting for the increased risk of disability from melanocyte malfunction, such as hearing and vision loss among people with elevated cadmium exposure.

  4. Effect of Xiaobailing Decoction on Melanocytes in Vitro

    Institute of Scientific and Technical Information of China (English)

    Xue Wenhui; Qu Xing; Chen Changbin; Li Minying

    2007-01-01

    Objective: To observe the effect of Xiaobailing Decoction ( 消白灵汤 ) on murine melanocytes in vitro and to explore the mechanism of Xiaobailing Decoction in the treatment of vitiligo. Methods: B-16F10murine melanoma cells were cultured in 1640 medium and treated respectively with different concentrations (1mg/ml, 2mg/ml, 3mg/ml) of the Chinese drug Xiaobailing Decoction and its main components, the drugs for replenishing the kidney-yang, and the drugs for nourishing blood and activating blood circulation, etc. For 7 days. MMT assay was used to determine the proliferation of B-16F10 murine melanoma cells. NaOH cleavage assay was adopted to measure the melanogenesis of melanocytes. Results:Xiaobailing Decoction, the drugs for replenishing the kidney-yang and the drugs for nourishing blood and activating blood circulation at different concentrations significantly improved the proliferation of B-16F10murine melanoma cells from the 3rd day to the 5th day (P<0.05), with Xiaobailing Decoction at the concentrations of 1mg/ml having the most distinct action on promoting the proliferation of melanocytes on the 3rd day (P<0.001); And the drugs for replenishing the kidney-yang at the concentrations of 2mg/ml and 3mg/ml and the drugs for nourishing blood and activating blood circulation at 3mg/ml significantly increased melanogenesis of melanocytes (P<0.05). Conclusion: Xiaobailing Decoction can promote melanocytic proliferation and melanogenesis in vitro, and it is indicated that the drugs for replenishing the kidney-yang and the drugs for nourishing blood and activating blood circulation play an important role in treating vitiligo.

  5. Melanocyte colonization and pigmentation of breast carcinoma

    DEFF Research Database (Denmark)

    Mele, Marco; Laurberg, Tinne; Engberg Damsgaard, Tine

    2012-01-01

    Introduction. Melanocyte colonization of breast carcinoma by nonneoplastic melanocytes of epidermal origin is a rare and serious condition first described in 1977. We report on the exceptional clinical and pathological features of this migration phenomenon in a 74-year-old patient. Discussion....... The pathogenesis by which melanocyte migration takes place is not known, but a breached basement membrane is considered essential. Conclusion. Histological examination and additional staining of skin are essential to differentiate breast cancer melanosis from malignant melanoma....

  6. Pleiotrophin inhibits melanogenesis via Erk1/2-MITF signaling in normal human melanocytes.

    Science.gov (United States)

    Choi, Woo Jong; Kim, Misun; Park, Ji-Youn; Park, Tae Jun; Kang, Hee Young

    2015-01-01

    Pleiotrophin (PTN) is a secreted heparin-binding protein that is involved in various biological functions of cell growth and differentiation. Little is known about the effects of PTN on the melanocyte function and skin pigmentation. In this study, we investigated whether PTN would affect melanogenesis. PTN was expressed in melanocytes and fibroblasts of human skin. Transfection studies revealed that PTN decreased melanogenesis, probably through MITF degradation via Erk1/2 activation in melanocytes. The inhibitory action of PTN in pigmentation was further confirmed in ex vivo cultured skin and in the melanocytes cocultured with fibroblasts. These findings suggest that PTN is a crucial factor for the regulation of melanogenesis in the skin.

  7. Modulation of Melanogenesis and Antioxidant Status of Melanocytes in Response to Phototoxic Action of Doxycycline.

    Science.gov (United States)

    Rok, Jakub; Buszman, Ewa; Beberok, Artur; Delijewski, Marcin; Otręba, Michał; Wrześniok, Dorota

    2015-11-01

    Doxycycline is a commonly used tetracycline antibiotic showing the broad spectrum of antibacterial action. However, the use of this antibiotic is often connected with the risk of phototoxic reactions that lead to various skin disorders. One of the factors influencing the photosensitivity reactions is the melanin content in melanocytes. In this study, the impact of doxycycline and UVA irradiation on cell viability, melanogenesis and antioxidant defense system in cultured normal human epidermal melanocytes (HEMn-DP) was examined. The exposure of cells to doxycycline and UVA radiation resulted in concentration-dependent loss in melanocytes viability and induced melanin biosynthesis. Significant changes were stated in cellular antioxidant enzymes activity: SOD, CAT and GPx, which indicates alterations of antioxidant defense system. The results obtained in vitro may explain the mechanisms of phototoxic reactions that occur in normal human epidermal melanocytes in vivo after exposure of skin to doxycycline and UVA radiation.

  8. Gibberellin production and plant growth promotion from pure cultures of Cladosporium sp. MH-6 isolated from cucumber (Cucumis sativus L.).

    Science.gov (United States)

    Hamayun, Muhammad; Khan, Sumera Afzal; Khan, Abdul Latif; Rehman, Gauhar; Kim, Youn-Ha; Iqbal, Ilyas; Hussain, Javid; Sohn, Eun-Young; Lee, In-Jung

    2010-01-01

    Gibberellin (GA) production by soil fungi has received little attention, although substantial work has been carried out on other aspects of plant growth promoting fungi (PGPF). In our studies we investigated GA production and growth-promoting capacity of a novel fungal strain isolated from the roots of soil-grown cucumber. Pure cultures of 19 endophytic fungi were tested for shoot length promotion of Waito-C rice to identify the GA production capacity of these fungal isolates. Isolate MH-6 significantly increased shoot length (12.9 cm) of Waito-C, in comparison to control treatments. Bioassay with culture filtrate (CF) of MH-6 also significantly promoted growth attributes of cucumber plants. Analysis of MH-6 CF showed the presence of physiologically active (GA1, 1.97 ng/mL; GA3, 5.18 ng/mL; GA4, 13.35 ng/mL and GA7, 2.4 ng/ mL) in conjunction with physiologically inactive (GA9 [0.69 ng/mL], GA12 [0.24 ng/mL], GA15 [0.68 ng/ mL, GA19 [1.94 ng/mL and GA20 [0.78 ng/mL]) gibberellins. The CF of MH-6 produced greater amounts of GA3, GA4, GA7 and GA19 than wild type Fusarium fujikuroi, a fungus known for high production of GA. The fungal isolate MH-6 was identified as a new strain of Cladosporium sp. on the basis of sequence homology (99%) and phylogenetic analysis of 18S rDNA sequence.

  9. Biomineralization of 1,4-dioxane in Pure Culture, Microcosm, and Column Studies Using 13C Labeling

    Science.gov (United States)

    Rolston, H. M.; Azizian, M.; Hyman, M. R.; Semprini, L.

    2016-12-01

    1,4-dioxane (1,4-D), a probable human carcinogen at low (biomineralizes 1,4-D to CO2. Batch experiments have been conducted with pure culture 21198 and in microcosms constructed with aquifer sediments. The rate of resting cell transformation of 1,4-D by ATCC 21198 was over 100 times faster than the rate of CO2 accumulation, indicating the presence of intermediates that were slowly mineralized to CO2 . In microcosms, the use of isobutane as a primary substrate effectively stimulated the native microbial community to transform 1,4-D. Microcosms were also bioaugmented with ATCC 21198. After an initial lag and subsequent additions of isobutane, transformation rates in the native microcosms approached those of the bioaugmented microcosms. Cometabolically active microbes survived several periods of starvation in all microcosms, and nutrient amendment allowed for sustained transformation rates. 13C labeled 1,4-D is currently being used to determine the rates and extents of biomineralization in the microcosms. Column studies are also being conducted to evaluate cometabolism and biominerazation potential of isobutane as a biostimulant and 21198 for bioaugmentation under geochemical and flow conditions more representative of in-situ bioremediation.

  10. The evaluation of kefir pure culture starter: Liquid-core capsule entrapping microorganisms isolated from kefir grains.

    Science.gov (United States)

    Wang, Liang; Zhong, Hao; Liu, Keying; Guo, Aizhen; Qi, Xianghui; Cai, Meihong

    2016-10-01

    The main purpose of this study was to develop a pure culture starter for producing kefir. In order to accomplish starter recycling, yeasts (Kluyveromyces marxianus strain, Pichia kudriavzevii clone), lactic acid bacteria (Lactobacillus kefiri strain F4Aa, Lactobacillus kefiri strain NM131-7, Lactobacillus kefiri strain NM132-3, Lactobacillus kefiri strain NM180-3, respectively), and acetic acid bacteria (Acetobacter lovaniensis strain) were entrapped in liquid core capsules based on the distribution ratio in kefir grains. The microbiological, antimicrobial, and chemical properties of kefir made with capsules (M) and kefir grains (K) were measured and compared. According to the results of plate counts in different selective medium, the number of yeasts and bacteria in the liquid core capsules gradually increased and stabilized after eight fermentation cycles. The results of gas chromatography-mass spectrometry showed that almost all the aroma components existed in the two type of kefir, except the ethyl lactate. There was no significant difference in alcohol content, protein content, and fat content, except the acidity and sugar content. Water holding capacity of kefir K was higher than kefir M. There were 14 same free amino acids in kefir M and kefir K, and the content of most free amino acids was similar. In antimicrobial test, there was no significant difference in both kefirs. © The Author(s) 2016.

  11. Oxidation of acetate through reactions of the citric acid cycle by Geobacter sulfurreducens in pure culture and in syntrophic coculture.

    Science.gov (United States)

    Galushko, A S; Schink, B

    2000-11-01

    Geobacter sulfurreducens strain PCA oxidized acetate to CO2 via citric acid cycle reactions during growth with acetate plus fumarate in pure culture, and with acetate plus nitrate in coculture with Wolinella succinogenes. Acetate was activated by succinyl-CoA:acetate CoA-transferase and also via acetate kinase plus phosphotransacetylase. Citrate was formed by citrate synthase. Soluble isocitrate and malate dehydrogenases NADP+ and NAD+, respectively. Oxidation of 2-oxoglutarate was measured as benzyl viologen reduction and strictly CoA-dependent; a low activity was also observed with NADP+. Succinate dehydrogenase and fumarate ductase both were membrane-bound. Succinate oxidation was coupled to NADP+ reduction whereas fumarate reduction was coupled to NADPH and NADH Coupling of succinate oxidation to NADP+ or cytochrome(s) reduction required an ATP-dependent reversed electron transport. Net ATP synthesis proceeded exclusively through electron transport phosphorylation. During fumarate reduction, both NADPH and NADH delivered reducing equivalents into the electron transport chain, which contained a menaquinone. Overall, acetate oxidation with fumarate proceeded through an open loop of citric acid cycle reactions, excluding succinate dehydrogenase, with fumarate reductase as the key reaction for electron delivery, whereas acetate oxidation in the syntrophic coculture required the complete citric acid cycle.

  12. Effects of Arsenite Resistance on the Growth and Functional Gene Expression of Leptospirillum ferriphilum and Acidithiobacillus thiooxidans in Pure Culture and Coculture

    Directory of Open Access Journals (Sweden)

    Huidan Jiang

    2015-01-01

    Full Text Available The response of iron-oxidizing Leptospirillum ferriphilum YSK and sulfur-oxidizing Acidithiobacillus thiooxidans A01 to arsenite under pure culture and coculture was investigated based on biochemical characterization (concentration of iron ion and pH value and related gene expression. L. ferriphilum YSK and At. thiooxidans A01 in pure culture could adapt up to 400 mM and 800 mM As(III after domestication, respectively, although arsenite showed a negative effect on both strains. The coculture showed a stronger sulfur and ferrous ion oxidation activity when exposed to arsenite. In coculture, the pH value showed no significant difference when under 500 mM arsenite stress, and the cell number of At. thiooxidans was higher than that in pure culture benefiting from the interaction with L. ferriphilum. The expression profile showed that the arsenic efflux system in the coculture was more active than that in pure culture, indicating that there is a synergetic interaction between At. thiooxidans A01 and L. ferriphilum YSK. In addition, a model was proposed to illustrate the interaction between arsenite and the ars operon in L. ferriphilum YSK and At. thiooxidans A01. This study will facilitate the effective application of coculture in the bioleaching process by taking advantage of strain-strain communication and coordination.

  13. Effects of Arsenite Resistance on the Growth and Functional Gene Expression of Leptospirillum ferriphilum and Acidithiobacillus thiooxidans in Pure Culture and Coculture.

    Science.gov (United States)

    Jiang, Huidan; Liang, Yili; Yin, Huaqun; Xiao, Yunhua; Guo, Xue; Xu, Ying; Hu, Qi; Liu, Hongwei; Liu, Xueduan

    2015-01-01

    The response of iron-oxidizing Leptospirillum ferriphilum YSK and sulfur-oxidizing Acidithiobacillus thiooxidans A01 to arsenite under pure culture and coculture was investigated based on biochemical characterization (concentration of iron ion and pH value) and related gene expression. L. ferriphilum YSK and At. thiooxidans A01 in pure culture could adapt up to 400 mM and 800 mM As(III) after domestication, respectively, although arsenite showed a negative effect on both strains. The coculture showed a stronger sulfur and ferrous ion oxidation activity when exposed to arsenite. In coculture, the pH value showed no significant difference when under 500 mM arsenite stress, and the cell number of At. thiooxidans was higher than that in pure culture benefiting from the interaction with L. ferriphilum. The expression profile showed that the arsenic efflux system in the coculture was more active than that in pure culture, indicating that there is a synergetic interaction between At. thiooxidans A01 and L. ferriphilum YSK. In addition, a model was proposed to illustrate the interaction between arsenite and the ars operon in L. ferriphilum YSK and At. thiooxidans A01. This study will facilitate the effective application of coculture in the bioleaching process by taking advantage of strain-strain communication and coordination.

  14. UVB-irradiated keratinocytes induce melanoma-associated ganglioside GD3 synthase gene in melanocytes via secretion of tumor necrosis factor α and interleukin 6

    Energy Technology Data Exchange (ETDEWEB)

    Miyata, Maiko [Department of Life and Medical Sciences, Chubu University Faculty of Life and Health Sciences, Matsumoto, Kasugai 487-8501 (Japan); Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Ichihara, Masatoshi; Tajima, Orie; Sobue, Sayaka; Kambe, Mariko [Department of Life and Medical Sciences, Chubu University Faculty of Life and Health Sciences, Matsumoto, Kasugai 487-8501 (Japan); Sugiura, Kazumitsu [Department of Dermatology, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Furukawa, Koichi, E-mail: koichi@med.nagoya-u.ac.jp [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Furukawa, Keiko [Department of Life and Medical Sciences, Chubu University Faculty of Life and Health Sciences, Matsumoto, Kasugai 487-8501 (Japan); Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan)

    2014-03-07

    Highlights: • Melanocytes showed low ST8SIA1 and high B3GALT4 levels in contrast with melanomas. • Direct UVB irradiation of melanocytes did not induce ganglioside synthase genes. • Culture supernatants of UVB-irradiated keratinocytes induced ST8SIA1 in melanocytes. • TNFα and IL-6 secreted from keratinocytes enhanced ST8SIA1 expression in melanocytes. • Inflammatory cytokines induced melanoma-related ST8SIA1 in melanocytes. - Abstract: Although expression of gangliosides and their synthetic enzyme genes in malignant melanomas has been well studied, that in normal melanocytes has been scarcely analyzed. In particular, changes in expression levels of glycosyltransferase genes responsible for ganglioside synthesis during evolution of melanomas from melanocytes are very important to understand roles of gangliosides in melanomas. Here, expression of glycosyltransferase genes related to the ganglioside synthesis was analyzed using RNAs from cultured melanocytes and melanoma cell lines. Quantitative RT-PCR revealed that melanomas expressed high levels of mRNA of GD3 synthase and GM2/GD2 synthase genes and low levels of GM1/GD1b synthase genes compared with melanocytes. As a representative exogenous stimulation, effects of ultraviolet B (UVB) on the expression levels of 3 major ganglioside synthase genes in melanocytes were analyzed. Although direct UVB irradiation of melanocytes caused no marked changes, culture supernatants of UVB-irradiated keratinocytes (HaCaT cells) induced definite up-regulation of GD3 synthase and GM2/GD2 synthase genes. Detailed examination of the supernatants revealed that inflammatory cytokines such as TNFα and IL-6 enhanced GD3 synthase gene expression. These results suggest that inflammatory cytokines secreted from UVB-irradiated keratinocytes induced melanoma-associated ganglioside synthase genes, proposing roles of skin microenvironment in the promotion of melanoma-like ganglioside profiles in melanocytes.

  15. Ultrastructure of Amelanotic Melanocytes from Human Hair Follicles

    Institute of Scientific and Technical Information of China (English)

    Ruzhi Zhang; Wenyuan Zhu; Mingyu Xia; Daguang Wang; Huijun Ma

    2007-01-01

    Objective: To investigate the ultra structure of amelanotic melanocytes (AMMC). Methods: The hair follicles obtained from normal human scalp by 0.50% collagenase type V treatment were washed with 0.1mol/L phosphate buffer salt (PBS). Hair-follicle cell suspensions were prepared by trypsin treatment and cultured in melanocyte medium. Remaining keratinocytes were removed by differential trypsinization. 100μg/ml geneticin was used to eliminate the contaminating fibroblasts. At third passage, the cells were trypsinized, and then washed in phosphate-buffered saline and processed for transmission electron microscopy. Results: Under transmission electron microscope, the cultured cells showed round or oval shape, with single large nuclear and the karyotheca were double deck. There were obvious euchromosome within the nucleus, and sparse heterochromosome. There were various organelles in the cytoplasm, including plentiful melanosomes with nearly similar size, mitochondria, rough endoplasmic reticule (RER) and ribosome. The electron density granules in most of the melanosomes disposed along concentric circularities. Golgi apparatus in the cells was inconspicuous. Conclusion: The ultra structure of AMMC from human hair follicles is different from that of epidermal melanocytes, and these characteristics determine the functional immature of AMMC.

  16. Substantial Effect of Melanin Influencing Factors on Melanogenesis in Muzzle Melanocytes of Differently Colored Hanwoo

    Directory of Open Access Journals (Sweden)

    Touseef Amna

    2012-07-01

    Full Text Available The present study was designed to investigate the effect of α-melanocyte-stimulating hormone (α-MSH, nitric oxide (NO and L-cysteine on melanin production and expression of related genes MC1R, Tyr, Tyrp-1 and Tyrp-2 in muzzle melanocytes of differently colored three native Hanwoo cattle. Muzzle samples were taken from black, brindle and brown Hanwoo and purified melanocytes were cultured with α-MSH, nitric oxide and L-cysteine at 100 nM, 50 µM and 0.07 mg/ml of media respectively. The amounts of total melanin, eumelanin and mRNA expression at Tyr, Tyrp-1, Tyrp-2 and MC1R levels were quantified. α-MSH and nitric oxide significantly increased (p<0.05 the amount of total melanin in black and brindle whereas eumelanin production in brown Hanwoo muzzle melanocytes. On the contrary, L-cysteine greatly (p<0.05 depressed the eumelanin production in black color but increased in brown. Simultaneously, up regulation of Tyr by nitric oxide and α-MSH and down regulation of Tyr, Tyrp-2 and MC1R genes by L-cysteine were observed in muzzle melanocytes of all three phenotypes. The results of this study revealed nitric oxide and α-MSH contribute hyper-pigmentation by enhancing eumelanogenesis whereas L-cysteine contributes to pheomelanin production in different colored Hanwoo muzzle melanocytes.

  17. Effect of nicotine on melanogenesis and antioxidant status in HEMn-LP melanocytes

    Energy Technology Data Exchange (ETDEWEB)

    Delijewski, Marcin; Beberok, Artur; Otręba, Michał; Wrześniok, Dorota; Rok, Jakub; Buszman, Ewa, E-mail: ebuszman@sum.edu.pl

    2014-10-15

    Nicotine is a natural ingredient of tobacco plants and is responsible for the addictive properties of tobacco. Nowadays nicotine is also commonly used as a form of smoking cessation therapy. It is suggested that nicotine may be accumulated in human tissues containing melanin. This may in turn affect biochemical processes in human cells producing melanin. The aim of this study was to examine the effect of nicotine on melanogenesis and antioxidant status in cultured normal human melanocytes HEMn-LP. Nicotine induced concentration-dependent loss in melanocytes viability. The value of EC{sub 50} was determined to be 7.43 mM. Nicotine inhibited a melanization process in human light pigmented melanocytes and caused alterations of antioxidant defense system. Significant changes in cellular antioxidant enzymes: superoxide dismutase and catalase activities and in hydrogen peroxide content were stated. The obtained results may explain a potential influence of nicotine on biochemical processes in melanocytes in vivo during long term exposition to nicotine. - Graphical abstract: Nicotine inhibits melanogenesis and induces oxidative stress in HEMn-LP melanocytes. - Highlights: • Nicotine induces concentration-dependent loss in melanocytes viability. • Nicotine in non-cytotoxic concentrations inhibits melanogenesis. • Nicotine in higher concentrations induces oxidative stress.

  18. Analysis of the effects of hydroquinone and arbutin on the differentiation of melanocytes.

    Science.gov (United States)

    Inoue, Yu; Hasegawa, Seiji; Yamada, Takaaki; Date, Yasushi; Mizutani, Hiroshi; Nakata, Satoru; Matsunaga, Kayoko; Akamatsu, Hirohiko

    2013-01-01

    Hydroquinone (HQ) is a chemical compound that inhibits the functions of melanocytes and has long been known for its skin-whitening effect. According to previous studies, the Tyrosinase (Tyr) activity inhibitory effect and melanocyte-specific cell toxicity are known depigmenting mechanisms; however, details of the underlying mechanisms are unknown. Arbutin (Arb) is also known for its Tyr activity inhibitory effect and is commonly used as a skin-whitening agent. However, the detailed depigmenting mechanism of Arb is also not yet fully understood. Few studies have attempted to elucidate the effects of HQ and Arb on undifferentiated melanocytes. In this study, we examined the effects of HQ and Arb throughout each stage of differentiation of melanocytes using a mouse embryonic stem cell (ESC) culture system to induce melanocytes. The results showed that HQ in particular downregulated the early stage of differentiation, in which neural crest cells were generated, and the late stage of differentiation, in which melanogenesis became active. On the other hand, Arb had no effect on the differentiation of melanocytes, and only suppressed melanogenesis by specifically suppressing elevations in Tyr expression in the late stage of differentiation.

  19. [Molecular diagnosis of melanocytic tumors].

    Science.gov (United States)

    Bauer, J

    2016-01-01

    Melanoma therapy has undergone a paradigm shift. Classic chemotherapies with poor treatment responses have been replaced by modern immune checkpoint blockades and targeted therapies with excellent responses. The latter require precise diagnosis of mutations in the melanoma genome as molecular targets for the small molecules. The diagnosis of melanomas has also been supplemented by molecular techniques. Differential diagnosis of melanoma and melanoma simulators such as atypical Spitz nevi can be supported by fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH). Here we review the indications and methods for molecular diagnosis of melanocytic tumors.

  20. Amides from Piper nigrum L. with dissimilar effects on melanocyte proliferation in-vitro.

    Science.gov (United States)

    Lin, Zhixiu; Liao, Yonghong; Venkatasamy, Radhakrishnan; Hider, Robert C; Soumyanath, Amala

    2007-04-01

    Melanocyte proliferation stimulants are of interest as potential treatments for the depigmentary skin disorder, vitiligo. Piper nigrum L. (Piperaceae) fruit (black pepper) water extract and its main alkaloid, piperine (1), promote melanocyte proliferation in-vitro. A crude chloroform extract of P. nigrum containing piperine was more stimulatory than an equivalent concentration of the pure compound, suggesting the presence of other active components. Piperine (1), guineensine (2), pipericide (3), N-feruloyltyramine (4) and N-isobutyl-2E, 4E-dodecadienamide (5) were isolated from the chloroform extract. Their activity was compared with piperine and with commercial piperlongumine (6) and safrole (7), and synthetically prepared piperettine (8), piperlonguminine (9) and 1-(3, 4-methylenedioxyphenyl)-decane (10). Compounds 6-10 either occur in P. nigrum or are structurally related. Compounds 1, 2, 3, 8 and 9 stimulated melanocyte proliferation, whereas 4, 5, 6, 7 and 10 did not. Comparison of structures suggests that the methylenedioxyphenyl function is essential for melanocyte stimulatory activity. Only those compounds also possessing an amide group were active, although the amino component of the amide group and chain linking it to the methylenedioxyphenyl group can vary. P. nigrum, therefore, contains several amides with the ability to stimulate melanocyte proliferation. This finding supports the traditional use of P. nigrum extracts in vitiligo and provides new lead compounds for drug development for this disease.

  1. Helium-neon laser irradiation stimulates migration and proliferation in melanocytes and induces repigmentation in segmental-type vitiligo.

    Science.gov (United States)

    Yu, Hsin-Su; Wu, Chieh-Shan; Yu, Chia-Li; Kao, Ying-Hsien; Chiou, Min-Hsi

    2003-01-01

    Low-energy helium-neon lasers (632.8 nm) have been employed in a variety of clinical treatments including vitiligo management. Light-mediated reaction to low-energy laser irradiation is referred to as biostimulation rather than a thermal effect. This study sought to determine the theoretical basis and clinical evidence for the effectiveness of helium-neon lasers in treating vitiligo. Cultured keratinocytes and fibroblasts were irradiated with 0.5-1.5 J per cm2 helium-neon laser radiation. The effects of the helium-neon laser on melanocyte growth and proliferation were investigated. The results of this in vitro study revealed a significant increase in basic fibroblast growth factor release from both keratinocytes and fibroblasts and a significant increase in nerve growth factor release from keratinocytes. Medium from helium-neon laser irradiated keratinocytes stimulated [3H]thymidine uptake and proliferation of cultured melanocytes. Furthermore, melanocyte migration was enhanced either directly by helium-neon laser irradiation or indirectly by the medium derived from helium-neon laser treated keratinocytes. Thirty patients with segmental-type vitiligo on the head and/or neck were enrolled in this study. Helium-neon laser light was administered locally at 3.0 J per cm2 with point stimulation once or twice weekly. The percentage of repigmented area was used for clinical evaluation of effectiveness. After an average of 16 treatment sessions, initial repigmentation was noticed. Marked repigmentation (>50%) was observed in 60% of patients with successive treatments. Basic fibroblast growth factor is a putative melanocyte growth factor, whereas nerve growth factor is a paracrine factor for melanocyte survival in the skin. Both nerve growth factor and basic fibroblast growth factor stimulate melanocyte migration. It is reasonable to propose that helium-neon laser irradiation clearly stimulates melanocyte migration and proliferation and mitogen release for melanocyte growth

  2. Nitric oxide enhances the sensitivity of alpaca melanocytes to respond to {alpha}-melanocyte-stimulating hormone by up-regulating melanocortin-1 receptor

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Yanjun; Cao, Jing; Wang, Haidong; Zhang, Jie; Zhu, Zhiwei; Bai, Rui; Hao, HuanQing; He, Xiaoyan; Fan, Ruiwen [College of Animal Science and Technology, Shanxi Agricultural University, 030801 Taigu, Shanxi (China); Dong, Changsheng, E-mail: cs_dong@sxau.edu.cn [College of Animal Science and Technology, Shanxi Agricultural University, 030801 Taigu, Shanxi (China)

    2010-06-11

    Nitric oxide (NO) and {alpha}-melanocyte-stimulating hormone ({alpha}-MSH) have been correlated with the synthesis of melanin. The NO-dependent signaling of cellular response to activate the hypothalamopituitary proopiomelanocortin system, thereby enhances the hypophysial secretion of {alpha}-MSH to stimulate {alpha}-MSH-receptor responsive cells. In this study we investigated whether an NO-induced pathway can enhance the ability of the melanocyte to respond to {alpha}-MSH on melanogenesis in alpaca skin melanocytes in vitro. It is important for us to know how to enhance the coat color of alpaca. We set up three groups for experiments using the third passage number of alpaca melanocytes: the control cultures were allowed a total of 5 days growth; the UV group cultures like the control group but the melanocytes were then irradiated everyday (once) with 312 mJ/cm{sup 2} of UVB; the UV + L-NAME group is the same as group UV but has the addition of 300 {mu}M L-NAME (every 6 h). To determine the inhibited effect of NO produce, NO produces were measured. To determine the effect of the NO to the key protein and gene of {alpha}-MSH pathway on melanogenesis, the key gene and protein of the {alpha}-MSH pathway were measured by quantitative real-time PCR and Western immunoblotting. The results provide exciting new evidence that NO can enhance {alpha}-MSH pathway in alpaca skin melanocytes by elevated MC1R. And we suggest that the NO pathway may more rapidly cause the synthesis of melanin in alpaca skin under UV, which at that time elevates the expression of MC1R and stimulates the keratinocytes to secrete {alpha}-MSH to enhance the {alpha}-MSH pathway on melanogenesis. This process will be of considerable interest in future studies.

  3. Melanocyte migration is influenced by E-cadherin-dependent adhesion of keratinocytes in both two- and three-dimensional in vitro wound models.

    Science.gov (United States)

    Keswell, Dheshnie; Kidson, Susan H; Davids, Lester M

    2015-02-01

    During wound healing, melanocytes are required to migrate into the wounded area that is still in the process of re-construction. The role and behaviour of melanocytes during this process is poorly understood, that is, whether melanocyte migration into the wound is keratinocyte-dependent or not. This paper attempts, through the use of both two- and three-dimensional in vitro models, to understand the role and behaviour of melanocytes during the process of wound healing. In addition, it sheds light on whether keratinocytes influence/contribute toward melanocyte migration and ultimately wound healing. Scratch assays were performed to analyse migration and Western blot analyses measured cellular E-cadherin expression. Immunohistochemistry was used to analyse the in vivo 3D wound healing effect. Scratch assays performed on co-cultures of melanocytes and keratinocytes demonstrated that melanocytes actively migrated, with the use of their dendrites, into the scratch ahead of the proliferating keratinocyte sheet. Migration of the melanocyte into the wound bed was accompanied by loss of attachment to keratinocytes at the wound front with concomitant downregulation of E-cadherin expression as observed through immunocytochemistry. This result suggests that, in vitro, melanocyte migration occurs independently of keratinocytes but that the migration is influenced by keratinocyte E-cadherin expression. We now demonstrate that melanocyte migration during re-pigmentation is an active process, and suggest that targeting of mechanisms involved in active melanocyte migration (e.g. the melanocyte dendrite) may enhance the re-pigmentation process. © 2014 International Federation for Cell Biology.

  4. Cytosolic dsDNA triggers apoptosis and pro-inflammatory cytokine production in normal human melanocytes.

    Science.gov (United States)

    Wang, Suiquan; Liu, Dongyin; Ning, Weixuan; Xu, Aie

    2015-04-01

    Considerable evidence implicates that viral infection might be a participant factor in the pathogenesis of vitiligo. However, it is still unclear how viral infection leads to the melanocyte destruction. To elucidate the effects of viral dsDNA on the viability and cytokine synthesis of normal human melanocytes and to explore the underlying mechanisms, primary cultured normal human melanocytes were transfected with poly(dA:dT). The results demonstrated that poly(dA:dT) triggered apoptosis instead of pyroptosis in melanocytes. Knocking down AIM2 or RIG-I by RNA interference partially reduced the poly(dA:dT)-induced LDH release, suggesting the involvement of both nucleic acid sensors in the process of melanocyte death. Poly(dA:dT) induced the expression of pro-inflammatory cytokine genes including IFN-β, TNF-α, IL-6 and IL-8 as well, whereas the pro-inflammatory cytokine production was suppressed by RIG-I siRNA, but not by AIM2 siRNA. Poly(dA:dT) treatment increased the phosphorylation of p38 and JNK and NFκB. Accordingly, NFκB inhibitor Bay 11-7082 and JNK inhibitor SP600125 blocked the induction of the cytokine genes except IFN-β. The production of IL6 and IL8 was also suppressed by p38 inhibitor SB203580. On the contrary, the Poly(dA:dT)-induced melanocyte death was only decreased by SP600125. This study provides the possible mechanism of melanocyte destruction and immuno-stimulation in vitiligo by innate immune response following viral infection.

  5. Melanocytic nevi in children: A clinical study

    Directory of Open Access Journals (Sweden)

    Senthilkumar M

    2006-01-01

    Full Text Available BACKGROUND: Melanocytic nevi show different presentations in children which have not much studied in India. PURPOSES: To document the incidence and pattern of melanocytic nevi in children. METHODS: A total of 4,256 paediatric cases attending the dermatology OPD, JIPMER during the study period from August 2002 to August 2004 were screened for melanocytic nevi. FINDINGS: Out of these, 41 (0.96% cases were found to have melanocytic nevi. The mean age of these cases was 1.4 years (range of 3 days to 14 years. Fourteen (34.1% of them were males and 27 (65.9% were females with male to female ratio of 1:1.9. Majority of these cases (32 cases, 78% were in the age group of 0-1 year. Thirty-seven (90.2% cases had single lesion and 4 (9.8% cases had multiple lesions. The size of the nevi varied from 1.5 cm to 20 cm in 40 (97.6% cases and more than 20 cm in 1 (2.4% case. The most common site of involvement was the back (32 cases, 78% followed by head and neck (6 cases, 14.6%. Out of the total of 41 cases, 30 (73.2% were Mongolian spots, 8 (19.5% were congenital melanocytic nevi, one case (2.4% each had giant congenital melanocytic nevus, nevus spilus and nevus of Ota. Four cases had other associated cutaneous disorders. These included ashleaf macules in 2 cases, epidermolysis bullosa simplex in 1 case and a solitary case had both cafι-au-lait macule and mixed hemangioma. The systemic associations included seizures (suspected neurocutaneous melanosis in a solitary case of congenital melanocytic nevi. CONCLUSIONS: The spectrum of melanocytic nevi seen in children in this study partially differs from this of abroad studies. LIMITATION: Unicentre hospital-based study which cannot be generalized.

  6. Thiocyanate degradation by pure and mixed cultures of microorganisms Degradação de tiocianato por culturas puras e mixtas

    Directory of Open Access Journals (Sweden)

    Elaine M. Souza-Fagundes

    2004-12-01

    Full Text Available A mixed culture and a pure bacterial strain (BMV8 were isolated from a bioreactor for thiocyanate treatment. Both cultures removed 5 mM of thiocyanate from the medium in 36 hours. The mixed culture was able to tolerate concentrations up to 60 mM. The efficiency of thiocyanate degradation decreased when the cells were immobilized.Uma cultura mixta e uma linhagem bacteriana pura foram isoladas de um bioreator para tratamento de tiocianato. As culturas removeram 5mM de tiocianato do meio em 36 horas. A cultura mixta foi capaz de tolerar concentrações superiores a 60mM. A eficiência da degradação de tiocianato diminuiu quando as células foram imobilizadas.

  7. Purification and growth of melanocortin 1 receptor (Mc1r)- defective primary murine melanocytes is dependent on stem cell factor (SFC) from keratinocyte-conditioned media.

    Science.gov (United States)

    Scott, Timothy L; Wakamatsu, Kazumasa; Ito, Shosuke; D'Orazio, John A

    2009-12-01

    The melanocortin 1 receptor (MC1R) is a transmembrane G(s)-coupled surface protein found on melanocytes that binds melanocyte-stimulating hormone and mediates activation of adenylyl cyclase and generation of the second messenger cyclic AMP (cAMP). MC1R regulates growth and differentiation of melanocytes and protects against carcinogenesis. Persons with loss-offunction polymorphisms of MC1R tend to be UV-sensitive (fair-skinned and with a poor tanning response) and are at high risk for melanoma. Mechanistic studies of the role of MC1R in melanocytic UV responses, however, have been hindered in part because Mc1r-defective primary murine melanocytes have been difficult to culture in vitro. Until now, effective growth of murine melanocytes has depended on cAMP stimulation with adenylyl cyclase-activating or phosphodiesterase-inhibiting agents. However, rescuing cAMP in the setting of defective MC1R signaling would be expected to confound experiments directly testing MC1R function on melanocytic UV responses. In this paper, we report a novel method of culturing primary murine melanocytes in the absence of pharmacologic cAMP stimulation by incorporating conditioned supernatants containing stem cell factor derived from primary keratinocytes. Importantly, this method seems to permit similar pigment expression by cultured melanocytes as that found in the skin of their parental murine strains. This novel approach will allow mechanistic investigation into MC1R's role in the protection against UV-mediated carcinogenesis and determination of the role of melanin pigment subtypes on UV-mediated melanocyte responses.

  8. Uncovering the Role of BMP Signaling in Melanocyte Development and Melanoma Tumorigenesis

    Science.gov (United States)

    2015-06-01

    loss of function studies in zebrafish embryos and mammalian cultured cells to determine if GDF6 antagonizes melanocyte development. In this task...W., Burns, C.E. and Zon. L.I. (2008). Transparent adult zebrafish as a tool for in vivo transplantation analysis. Cell Stem Cell, 2, 183-9. 7

  9. "Melanocytic Nests Arising in Lichenoid Inflammation": Reappraisal of the Terminology "Melanocytic Pseudonests".

    Science.gov (United States)

    Chung, Hye Jin; Simkin, A David; Bhawan, Jag; Wolpowitz, Deon

    2015-12-01

    Pseudonests or pseudomelanocytic nests represent aggregates of cells and cell fragments, including keratinocytes, macrophages, lymphocytes, and occasional melanocytes. Pseudomelanocytic nests in the setting of lichenoid inflammation can mimic atypical melanocytic proliferations. Several reports documented nonspecific staining of pseudonests with melanoma antigen recognized by T cells-1/Melan-A, which can be detected in the cytoplasm of nonmelanocytic cells. In contrast, nuclear stains, such as MITF and SOX10, avoid this nonmelanocyte cytoplasmic staining. The authors have previously proposed the term melanocytic pseudonests to describe junctional nests with numerous (>2) true melanoma antigen recognized by T cells-1/Melan-A, SOX10, and MITF in a nonmelanocytic lesion with lichenoid inflammation (unilateral lichen planus pigmentosus/erythema dyschromicum perstans). In this study, the authors report another case of this phenomenon arising in a different lichenoid inflammatory dermatitis (lichen planus). The immunophenotype and number of clustered true melanocytes indicate that these dermoepidermal aggregates represent true melanocytic nests and not pseudonests of any type. Therefore, the authors propose the revised terminology of "melanocytic nests arising in lichenoid inflammation" to describe this novel pattern of benign melanocytic reorganization or proliferation in a subset of lichenoid dermatitides. Because this phenomenon can mimic atypical melanocytic proliferations, clinicopathologic correlation is essential for the correct diagnosis.

  10. Impact of kanamycin on melanogenesis and antioxidant enzymes activity in melanocytes--an in vitro study.

    Science.gov (United States)

    Wrześniok, Dorota; Otręba, Michał; Beberok, Artur; Buszman, Ewa

    2013-12-01

    Aminoglycosides, broad spectrum aminoglycoside antibiotics, are used in various infections therapy due to their good antimicrobial characteristics. However, their adverse effects such as nephrotoxicity and auditory ototoxicity, as well as some toxic effects directed to pigmented tissues, complicate the use of these agents. This study was undertaken to investigate the effect of aminoglycoside antibiotic-kanamycin on viability, melanogenesis and antioxidant enzymes activity in cultured human normal melanocytes (HEMa-LP). It has been demonstrated that kanamycin induces concentration-dependent loss in melanocytes viability. The value of EC50 was found to be ~6.0 mM. Kanamycin suppressed melanin biosynthesis: antibiotic was shown to inhibit cellular tyrosinase activity and to reduce melanin content in normal human melanocytes. Significant changes in the cellular antioxidant enzymes: SOD, CAT and GPx were stated in melanocytes exposed to kanamycin. Moreover, it was observed that kanamycin caused depletion of antioxidant defense sytem. It is concluded that the inhibitory effect of kanamycin on melanogenesis and not sufficient antioxidant defense mechanism in melanocytes in vitro may explain the potential mechanisms of undesirable side effects of this drug directed to pigmented tissues in vivo. © 2013 Wiley Periodicals, Inc.

  11. Nicotine impact on melanogenesis and antioxidant defense system in HEMn-DP melanocytes.

    Science.gov (United States)

    Delijewski, Marcin; Wrześniok, Dorota; Otręba, Michał; Beberok, Artur; Rok, Jakub; Buszman, Ewa

    2014-10-01

    Nicotine is a compound of tobacco plants and is responsible for addictive properties of tobacco which is used by about one billion of smokers all over the world. Recently, nicotine has drawn even more attention due to its presumed neuroprotective and antioxidant features as far as common use in various forms of smoking cessation therapies. It is suggested that nicotine may be accumulated in human tissues containing melanin. This may in turn influence biochemical processes in human cells producing melanin. The aim of this study was to examine the impact of nicotine on melanogenesis and antioxidant defense system in cultured normal human melanocytes (HEMn-DP). Nicotine induced concentration-dependent loss in melanocytes viability. The value of EC50 was determined to be 2.52 mM. Nicotine modulated melanin biosynthesis in normal human melanocytes. Significant changes in hydrogen peroxide content and cellular antioxidant enzymes: SOD, CAT, and GPx activities were stated in melanocytes exposed to nicotine, which indicates alterations of antioxidant defense system. The results obtained in vitro may explain a potential influence of nicotine on biochemical processes in melanocytes in vivo during long-term exposition to nicotine.

  12. Manassantin B inhibits melanosome transport in melanocytes by disrupting the melanophilin-myosin Va interaction.

    Science.gov (United States)

    Chang, Huikyoung; Choi, Hyunjung; Joo, Kyung-Mi; Kim, Daegun; Lee, Tae Ryong

    2012-11-01

    Human skin hyperpigmentation disorders occur when the synthesis and/or distribution of melanin increases. The distribution of melanin in the skin is achieved by melanosome transport and transfer. The transport of melanosomes, the organelles where melanin is made, in a melanocyte precedes the transfer of the melanosomes to a keratinocyte. Therefore, hyperpigmentation can be regulated by decreasing melanosome transport. In this study, we found that an extract of Saururus chinensis Baill (ESCB) and one of its components, manassantin B, inhibited melanosome transport in Melan-a melanocytes and normal human melanocytes (NHMs). Manassantin B disturbed melanosome transport by disrupting the interaction between melanophilin and myosin Va. Manassantin B is neither a direct nor an indirect inhibitor of tyrosinase. The total melanin content was not reduced when melanosome transport was inhibited in a Melan-a melanocyte monoculture by manassantin B. Manassantin B decreased melanin content only when Melan-a melanocytes were co-cultured with SP-1 keratinocytes or stimulated by α-MSH. Therefore, we propose that specific inhibitors of melanosome transport, such as manassantin B, are potential candidate or lead compounds for the development of agents to treat undesirable hyperpigmentation of the skin.

  13. EFFECTS OF BU GU ZHI(PSORALEA CORYLIFOLIA L) AND BAI ZHI (DAHURIAN ANGELICA ROOT) EXTRACTS ON MELANOCYTE ADHESION AND MIGRATION

    Institute of Scientific and Technical Information of China (English)

    Ma Huiqun; Feng Jie; Zhang Xianqi; Mu Kuanhou; Liu Chao; Niu Xinwu; Dang Qianli

    2005-01-01

    Objective To find direct effect of Chinese herbs Bu Gu Zhi (Psoralea corylifolia L) and Bai Zhi (Dahurian angelica root) Extracts on melanocyte adhesion and migration in vitro. Methods Ethanol extracts obtained from two kind of Chinese medicinable herbs were tested. Human melanocytes were obtained from neonatal foreskins and 48-well culture dish covered with fibronectin were used to perform melanocyte adhesion assay; Motility was assessed using the micropore filter method. Results: The extracts of Bu Gu Zhi(Psoralea corylifolia L), Bai Zhi(Dahurian angelica root) obviously showed an effect in increasing of human melanocyte adhesion and migration on fibronectin. Conclusion It is suggested that Buguzhi(Fructus Psoraleae) and Baizhi(Radix Angelicae Dahuricae) might induce melanocyte adhesion and/or migration in the treatment of vitiligo.

  14. Melanocyte and Melanoma Cell Activation by Calprotectin

    Directory of Open Access Journals (Sweden)

    Stephanie H. Shirley

    2014-01-01

    Full Text Available Calprotectin, a heterodimer of S100A8 and S100A9, is a proinflammatory cytokine released from ultraviolet radiation-exposed keratinocytes. Calprotectin binds to Toll-like receptor 4, the receptor for advanced glycation end-products, and extracellular matrix metalloproteinase inducer on target cells to stimulate migration. Melanocytes and melanoma cells produce little if any calprotectin, but they do express receptors for the cytokine. Thus, keratinocyte-derived calprotectin has the potential to activate melanocytes and melanoma cells within the epidermis in a paracrine manner. We examined the ability of calprotectin to stimulate proliferation and migration in normal human melanocytes and melanoma cells in vitro. We first showed, by immunofluorescence and quantitative RT-PCR, that the melanocytic cells employed expressed a calprotectin receptor, the receptor for advanced end-products. We then demonstrated that calprotectin significantly enhanced proliferation, migration, and Matrigel invasion in both normal human melanocytes and melanoma cells. Thus, calprotectin is one of the numerous paracrine factors released by ultraviolet radiation-exposed keratinocytes that may promote melanomagenesis and is a potential target for melanoma prevention or therapy.

  15. Biodegradation potential of pure and mixed bacterial cultures for removal of 4-nitroaniline from textile dye wastewater.

    Science.gov (United States)

    Khalid, Azeem; Arshad, Muhammad; Crowley, David E

    2009-03-01

    Environmentally toxic aromatic amines including nitroanilines are commonly generated in dye contaminated wastewater in which azo dyes undergo degradation under anaerobic conditions. The aim of this study was to develop a process for biological treatment of 4-nitroaniline. Three bacteria identified as Acinetobacter sp., Citrobacter freundii and Klebsiella oxytoca were isolated from enrichment cultures of activated sludge on 4-nitroaniline, after which the isolates and the mixed culture were studied to determine optimal conditions for biodegradation. HPLC analyses showed the mixed culture was capable of complete removal of 100micromol/L of 4-nitroaniline within 72h under aerobic conditions. There was an inverse linear relationship (R(2)=0.96) between the rate of degradation (V) and 4-nitraoaniline concentrations [S] over 100-1000micromol/L. The bacterial culture was also capable of decolorizing structurally different azo dyes (Acid Red-88, Reactive Black-5, Direct Red-81, and Disperse Orange-3) and also degraded nitrobenzene. Our findings show that enrichment cultures from activated sludge can be effective for the removal of dyes and their toxic intermediates, and that treatment may best be accomplished using an anaerobic-aerobic process.

  16. Links between Schwann cells and melanocytes in development and disease.

    Science.gov (United States)

    Van Raamsdonk, Catherine D; Deo, Mugdha

    2013-09-01

    Melanocytes are pigment-producing cells that reside in the skin, eyes, ears, heart, and central nervous system meninges of mammals. Schwann cells are glial cells, which closely associate with peripheral nerves, myelinating, and sheathing them. Melanocytes and Schwann cells both arise from the neural crest during development, and some melanocytes arise directly from Schwann cell precursors lining developing spinal nerves. In this review, we explore the connections between melanocytes and Schwann cells in development and transformation.

  17. Effects of pure starter cultures on physico-chemical and sensory quality of dry fermented Chinese-style sausage.

    Science.gov (United States)

    Rai, Krishna P; Zhang, Chunhui; Xia, Wen Shui

    2010-03-01

    Dry fermented Chinese-style sausages prepared in laboratory inoculating with Lactobacillus casei subsp. casei-1.001, Pediococcus pentosaceus-ATCC 33316, Staphylococcus xylosus-12 and without starter culture randomly sampled at 0, 3, 10, and 24 days of ripening were analyzed for physico-chemical and sensory qualities. A significant (psausage during ripening was observed, whereas other major chemical parameters remained unchanged. The microbial fermentation resulted in decreased pH and nitrite but increased non protein nitrogen and total volatile basic nitrogen in the products. Starter cultures except P. Pentosaceous-ATCC 33316, used in the sausage failed to suppress rancidity in ripened product as indicated by a significant (psausages decreased with ripening time, meanwhile the redness (a) increased significantly (psausages inoculated with cultures L. casei subsp. casei-1.001 and S. xylosus-12. The texture profile of sausages was almost similar except for P. Pentosaceous-ATCC 33316, which showed significantly (psausage of high quality.

  18. Exploring characteristics of bioelectricity generation and dye decolorization of mixed and pure bacterial cultures from wine-bearing wastewater treatment.

    Science.gov (United States)

    Han, Jing-Long; Liu, Ying; Chang, Chang-Tang; Chen, Bor-Yann; Chen, Wen-Ming; Xu, Hui-Zhong

    2011-04-01

    This study uncovered microbial characteristics of bioelectricity generation and dye decolorization in single-chamber microbial fuel cells (MFCs) using activated sludge for wine-containing wastewater treatment. Phylogenetic tree analysis on 16S rRNA gene fragments indicated that the predominant strains on anodic biofilm in acclimatized MFCs were Gamma-Proteobacteria Aeromonas punctata NIU-P9, Pseudomonas plecoglossicida NIU-Y3, Pseudomonas koreensis NIU-X8, Acinetobacter junii NIU-Y8, Stenotrophomonas maltophila NIU-X2. Our findings showed that the current production capabilities of these pure strains were only ca. 10% of those of their mother activated sludge, indicating that synergistic interactions among microbes might be the most influential factor to maximize power generation in MFCs. Plus, these electrochemically active strains also performed reductive decolorization of C.I. reactive blue 160, suggesting that bioelectricity generation might be directly associated to azo dye decolorization to deal with electron transfer on anodic biofilm in MFCs.

  19. Production of NO and N2O by Pure Cultures of Nitrifying and Denitrifying Bacteria during Changes in Aeration

    NARCIS (Netherlands)

    Kester, R.A.; Boer, W. de; Laanbroek, H.J.

    1997-01-01

    Peak emissions of NO and N2O are often observed after wetting of soil. The reactions to sudden changes in the aeration of cultures of nitrifying and denitrifying bacteria with respect to NO and N2O emissions were compared to obtain more information about the microbiological aspects of peak emissions

  20. PAX3 expression in normal skin melanocytes and melanocytic lesions (naevi and melanomas.

    Directory of Open Access Journals (Sweden)

    Sandra Medic

    Full Text Available BACKGROUND: Cutaneous Malignant Melanoma is an aggressive form of skin cancer, arising in cutaneous melanocytes. The transcription factor PAX3 regulates melanocyte specification from neural crest cells during development but expression in differentiated melanocytes is uncertain. By contrast it is frequently found in melanomas and naevi and is a marker for melanoma staging and detection. In this study we analysed the expression of PAX3 across the spectrum of melanocytic cells, from normal melanocytes to cells of benign and malignant lesions to better assess its function in these various tissues. Pax3 and PAX3 (italicized refer to the mouse and human gene, respectively; whereas Pax3 and PAX3 (non-italicized refer to the corresponding mouse and human protein. METHODOLOGY AND PRINCIPAL FINDINGS: PAX3 expression was analysed by immunohistochemistry and qRT-PCR. Immunofluorescence was used for co-expression with differentiation, migration and survival markers. As expected PAX3 expression was observed in naevi and melanoma cells. It was also found in melanocytes of normal skin where it co-expressed with melanocyte markers, MITF and MLANA. Co-expression with its downstream target, antiapoptotic factor BCL2L1 confirms PAX3 as a cell survival regulator. PAX3 was also co-expressed with melanoma cell migration marker MCAM in dermal naevi and melanoma cell nests, but this downstream target of PAX3 was not present in normal epidermal melanocytes, suggesting differential roles for PAX3 in normal epidermal melanocytes and melanoma cells. Most interestingly, a proportion of PAX3-positive epidermal melanocytes in normal skin show HES1 and Ki67 co-expression, indicating their less differentiated proliferative phenotype. CONCLUSIONS AND SIGNIFICANCE: Our results suggest that a previously identified role for PAX3, that of regulator of an undifferentiated plastic state, may operate in melanocytes of normal skin. This role, possibly required for cellular response to

  1. Differential responses of normal human melanocytes to intra- and extracellular dsRNA.

    Science.gov (United States)

    Wang, Suiquan; Liu, Dongyin; Jin, Rong; Zhu, Yiping; Xu, Aie

    2015-06-01

    Viral factor has been implicated in the etiopathogenesis of vitiligo. To elucidate the effects of viral double-stranded RNA (dsRNA) on melanocytes and to explore the underlying mechanisms, primary cultured normal human melanocytes were treated with synthetic viral dsRNA analog poly(I:C). The results demonstrated that poly(I:C)-triggered apoptosis when transfected into melanocytes, while extracellular poly(I:C) did not have that effect. Intracellular poly(I:C)-induced melanocyte death was decreased by RIG-I or MDA5 siRNA, but not by TLR3 siRNA. Both intracellular and extracellular poly(I:C) induced the expression of IFNB, TNF, IL6, and IL8. However, extracellular poly(I:C) demonstrated a much weaker induction capacity of cytokine genes than intracellular poly(I:C). Further analysis revealed that phosphorylation of TBK1, IRF3, IRF7, and TAK1 was differentially induced by intra- or extracellular poly(I:C). NFκB inhibitor Bay 11-7082 decreased the induction of all the cytokines by poly(I:C), suggesting the ubiquitous role of NFκB in the process. Poly(I:C) treatment also induced the phosphorylation of p38 and JNK in melanocytes. Both JNK and p38 inhibitors showed suppression on the cytokine induction by intra- or extracellular poly(I:C). However, only the JNK inhibitor decreased the intracellular poly(I:C)-induced melanocyte death. Taken together, this study provides the possible mechanism of viral factor in the pathogenesis of vitiligo.

  2. UVB-irradiated keratinocytes induce melanoma-associated ganglioside GD3 synthase gene in melanocytes via secretion of tumor necrosis factor α and interleukin 6.

    Science.gov (United States)

    Miyata, Maiko; Ichihara, Masatoshi; Tajima, Orie; Sobue, Sayaka; Kambe, Mariko; Sugiura, Kazumitsu; Furukawa, Koichi; Furukawa, Keiko

    2014-03-07

    Although expression of gangliosides and their synthetic enzyme genes in malignant melanomas has been well studied, that in normal melanocytes has been scarcely analyzed. In particular, changes in expression levels of glycosyltransferase genes responsible for ganglioside synthesis during evolution of melanomas from melanocytes are very important to understand roles of gangliosides in melanomas. Here, expression of glycosyltransferase genes related to the ganglioside synthesis was analyzed using RNAs from cultured melanocytes and melanoma cell lines. Quantitative RT-PCR revealed that melanomas expressed high levels of mRNA of GD3 synthase and GM2/GD2 synthase genes and low levels of GM1/GD1b synthase genes compared with melanocytes. As a representative exogenous stimulation, effects of ultraviolet B (UVB) on the expression levels of 3 major ganglioside synthase genes in melanocytes were analyzed. Although direct UVB irradiation of melanocytes caused no marked changes, culture supernatants of UVB-irradiated keratinocytes (HaCaT cells) induced definite up-regulation of GD3 synthase and GM2/GD2 synthase genes. Detailed examination of the supernatants revealed that inflammatory cytokines such as TNFα and IL-6 enhanced GD3 synthase gene expression. These results suggest that inflammatory cytokines secreted from UVB-irradiated keratinocytes induced melanoma-associated ganglioside synthase genes, proposing roles of skin microenvironment in the promotion of melanoma-like ganglioside profiles in melanocytes.

  3. The Effect of Otic Melanocyte Destruction on Auditory and Vestibular Function: a Study on Vitiligo Patients

    Directory of Open Access Journals (Sweden)

    Parvane Mahdi

    2016-03-01

    Full Text Available The hallmark of vitiligo is the disappearance of melanocytes from the skin. As a result, of melanocytes presence in the auditory and vestibular apparatus, the involvement of these systems in vitiligo which targets the melanocytes of the whole body is possible; suggesting that vitiligo is a systemic disease rather than a purely cutaneous problem. A total of 21 patients with vitiligo were enrolled in this study. A group of 20 healthy subjects served as a control group. Pure tone audiometry (PTA, auditory brainstem responses (ABR and vestibular evoked myogenic potentials (VEMP were carried out in all participants. High frequency sensory neural hearing loss was seen in 8 (38.09% patients. ABR analysis revealed 10 (47.61% had an abnormal increase in latency of wave III, and 6 (28.57% had an abnormal prolongation of IPL I-III, however, regarding our VEMP findings, there were no recorded responses on left ear of 1 (4.76% patient and latency of p13 was prolonged in 5(23.80% patients. There was no correlation between ages, duration of disease, and any of the recorded parameters (P>0.05. In the present survey, we highlighted the auditory and vestibular involvement in vitiligo patients.

  4. Pure populations of murine macrophages from cultured embryonic stem cells. Application to studies of chemotaxis and apoptotic cell clearance.

    Science.gov (United States)

    Zhuang, Lihui; Pound, John D; Willems, Jorine J L P; Taylor, A Helen; Forrester, Lesley M; Gregory, Christopher D

    2012-11-30

    Embryonic stem cells provide a potentially convenient source of macrophages in the laboratory. Given the propensity of macrophages for plasticity in phenotype and function, standardised culture and differentiation protocols are required to ensure consistency in population output and activity in functional assays. Here we detail the development of an optimised culture protocol for the production of murine embryonic stem cell-derived macrophages (ESDM). This protocol provides improved yields of ESDM and we demonstrate that the cells are suitable for application to the study of macrophage responses to apoptotic cells. ESDM so produced were of higher purity than commonly used primary macrophage preparations and were functional in chemotaxis assays and in phagocytosis of apoptotic cells. Maturation of ESDM was found to be associated with reduced capacity for directed migration and increased capacity for phagocytic clearance of apoptotic cells. These results show ESDM to be functionally active in sequential phases of interaction with apoptotic cells and establish these macrophage populations as useful models for further study of molecular mechanisms underlying the recognition and removal of apoptotic cells. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. 纯菌种培养红茶菌中细菌纤维素的合成%Bacterial cellulose in Kombucha by pure cultures

    Institute of Scientific and Technical Information of China (English)

    周艳; 谭丽丽; 唐欣昀

    2012-01-01

    实验采用纯菌种培养红茶菌研究细菌纤维素(bacterial cellulose,BC)的合成量及产率,所用的菌株为汉逊氏葡糖酸醋杆菌(Gluconacetobacter hansenii CGMCC1671)和啤酒酵母(Saccharomyces cerevisiae CGMCC1670),所用培养基为糖茶水培养基。研究了碳源、茶叶种类、茶叶用量、种龄、温度和装液量等6个因素对红茶菌合成BC的影响。静止培养22d后测BC的产量和产率。结果表明红茶菌合成BC的最佳碳源为葡萄糖,最佳茶叶种类为绿茶,最佳茶叶用量为4g/L,最佳种龄为60h,最佳温度为30℃,最佳装液量为250mL的三角瓶中装75mL的培养液。这些最佳参数和红茶菌的培养相一致。纯菌种培养红茶菌技术可以应用于生产高品质及高产量的BC和高品质的红茶菌饮料。%Strains Gluconacetobacter hansenii CGMCC1671 and Saccharomyces cerevisiae CGMCC1670 were applied to make traditional Kombucha with pure cultures to search for the optimum parameters of major factors affecting the yields and productivities of bacterial cellulose(BC) in the beverage.Six culture factors were examined:carbon sources,tea kind,tea dosage,inoculum age,temperature and culture volume.The yields and productivities of BC and sugar consumed were measured after cultured statically for 22d.Glucose at 50g/L,green tea at 4g/L,inoculum of 60h,30℃ and 75mL were the optimum conditions for high yield of BC.These optimum parameters were nearly equal for both the maturation of Kombucha and the BC yields.The co-culture of pure strains of traditional Kombucha technique can be used to provide both high quality and high yield of BC in addition to producing high quality Kombucha beverage.

  6. Comparative genomic analysis of the gut bacterium Bifidobacterium longum reveals loci susceptible to deletion during pure culture growth

    Directory of Open Access Journals (Sweden)

    Shakhova VV

    2008-05-01

    Full Text Available Abstract Background Bifidobacteria are frequently proposed to be associated with good intestinal health primarily because of their overriding dominance in the feces of breast fed infants. However, clinical feeding studies with exogenous bifidobacteria show they don't remain in the intestine, suggesting they may lose competitive fitness when grown outside the gut. Results To further the understanding of genetic attenuation that may be occurring in bifidobacteria cultures, we obtained the complete genome sequence of an intestinal isolate, Bifidobacterium longum DJO10A that was minimally cultured in the laboratory, and compared it to that of a culture collection strain, B. longum NCC2705. This comparison revealed colinear genomes that exhibited high sequence identity, except for the presence of 17 unique DNA regions in strain DJO10A and six in strain NCC2705. While the majority of these unique regions encoded proteins of diverse function, eight from the DJO10A genome and one from NCC2705, encoded gene clusters predicted to be involved in diverse traits pertinent to the human intestinal environment, specifically oligosaccharide and polyol utilization, arsenic resistance and lantibiotic production. Seven of these unique regions were suggested by a base deviation index analysis to have been precisely deleted from strain NCC2705 and this is substantiated by a DNA remnant from within one of the regions still remaining in the genome of NCC2705 at the same locus. This targeted loss of genomic regions was experimentally validated when growth of the intestinal B. longum in the laboratory for 1,000 generations resulted in two large deletions, one in a lantibiotic encoding region, analogous to a predicted deletion event for NCC2705. A simulated fecal growth study showed a significant reduced competitive ability of this deletion strain against Clostridium difficile and E. coli. The deleted region was between two IS30 elements which were experimentally

  7. DMEM enhances tyrosinase activity in B16 mouse melanoma cells and human melanocytes

    Directory of Open Access Journals (Sweden)

    Panpen Diawpanich

    2008-07-01

    Full Text Available Media components may affect the activities of cultured cells. In this study, tyrosinase activity was evaluated by using B16-F10 mouse melanoma cell lines (B16-F10 and primary human melanocytes cultured in different media. An optical density measurement and a L-dopa reaction assay were used as the determination of the tyrosinase activity. The study of B16-F10 found the optical density to be 2010, 2246 and 2961 in cells cultured in RPMI Medium 1640 (RPMI1640,Minimum Essential Medium (MEM and Dulbecco’s Modified Eagle Medium (DMEM, respectively. Moreover, compared to RPMI 1640 and MEM, DMEM showed the darkest color of melanin formation in culture media and in cells after the L-dopa reaction assay. Addition of kojic acid showed a significant inhibitory effect on tyrosinase activity in all media.Whereas MCDB153 showed no significant effect on human melanocytes, DMEM caused a dramatic increase in tyrosinase activity after 4 days of cultivation. Addition of kojic acid showed a significant tyrosinase inhibitory effect in DMEM only. Furthermore, an active ingredient in green tea, epigallocathechin gallate (EGCG could inhibit tyrosinase activity in both B16-F10 and human melanocytes cultured in DMEM. In summary, these results suggest that DMEM is a suitable medium that provides high detection sensitivity in a tyrosinase inhibition assay.

  8. Clear-cell melanocytic lesions with balloon-cell and sebocyte-like melanocytes: a unifying concept.

    Science.gov (United States)

    Kazlouskaya, Viktoryia; Guo, Ying; Maia-Cohen, Sandra; Mones, Joan

    2014-05-01

    Melanocytes may assume unique shapes and sizes but rarely have clear cytoplasm. We studied 28 melanocytic lesions that contained clear-cell melanocytes of the balloon-cell and sebocyte-like types. Clear-cell melanocytes were found more commonly in females (64%) than in males (36%), with predominance in females younger than 50 years (79%) and predominance in males older than 50 years (67%). They were distributed evenly throughout the body but were not found on acral sites. Clear-cell melanocytes were most prevalent in congenital nevi (18 or 72%) but were also found in 5 Clark nevi, 2 Meischer nevi, 1 Unna nevus, 1 atypical intra-epidermal proliferation, and 1 melanoma. The clear cells were distributed diffusely in 86% of the lesions and focally in 14%. The overall percentage of clear-cell melanocytes was 56%. The percentage of balloon cells was 57% and sebocyte-like melanocytes 32%. Clear cells with morphologic features of both balloon cells and sebocyte-like melanocytes, that is, intermediate cells, were present in all lesions with an overall percentage of 12%. The presence of melanocytes of both the balloon-cell and sebocyte-like types and the finding of clear-cell melanocytes with intermediate features in all lesions lends support to the theory that balloon-cell and sebocyte-like melanocytes may represent morphologic expressions of the same alteration in melanogenesis.

  9. Melanocyte antigen triggers autoimmunity in human psoriasis.

    Science.gov (United States)

    Arakawa, Akiko; Siewert, Katherina; Stöhr, Julia; Besgen, Petra; Kim, Song-Min; Rühl, Geraldine; Nickel, Jens; Vollmer, Sigrid; Thomas, Peter; Krebs, Stefan; Pinkert, Stefan; Spannagl, Michael; Held, Kathrin; Kammerbauer, Claudia; Besch, Robert; Dornmair, Klaus; Prinz, Jörg C

    2015-12-14

    Psoriasis vulgaris is a common T cell-mediated inflammatory skin disease with a suspected autoimmune pathogenesis. The human leukocyte antigen (HLA) class I allele, HLA-C*06:02, is the main psoriasis risk gene. Epidermal CD8(+) T cells are essential for psoriasis development. Functional implications of HLA-C*06:02 and mechanisms of lesional T cell activation in psoriasis, however, remained elusive. Here we identify melanocytes as skin-specific target cells of an HLA-C*06:02-restricted psoriatic T cell response. We found that a Vα3S1/Vβ13S1 T cell receptor (TCR), which we had reconstituted from an epidermal CD8(+) T cell clone of an HLA-C*06:02-positive psoriasis patient specifically recognizes HLA-C*06:02-positive melanocytes. Through peptide library screening, we identified ADAMTS-like protein 5 (ADAMTSL5) as an HLA-C*06:02-presented melanocytic autoantigen of the Vα3S1/Vβ13S1 TCR. Consistent with the Vα3S1/Vβ13S1-TCR reactivity, we observed numerous CD8(+) T cells in psoriasis lesions attacking melanocytes, the only epidermal cells expressing ADAMTSL5. Furthermore, ADAMTSL5 stimulation induced the psoriasis signature cytokine, IL-17A, in CD8(+) T cells from psoriasis patients only, supporting a role as psoriatic autoantigen. This unbiased analysis of a TCR obtained directly from tissue-infiltrating CD8(+) T cells reveals that in psoriasis HLA-C*06:02 directs an autoimmune response against melanocytes through autoantigen presentation. We propose that HLA-C*06:02 may predispose to psoriasis via this newly identified autoimmune pathway.

  10. Susceptibility to antibiotics of Vibrio sp. AO1 growing in pure culture or in association with its hydroid host Aglaophenia octodonta (Cnidaria, Hydrozoa).

    Science.gov (United States)

    Stabili, Loredana; Gravili, Cinzia; Boero, Ferdinando; Tredici, Salvatore M; Alifano, Pietro

    2010-04-01

    Vibrio harveyi is the major causal organism of vibriosis, causing potential devastation to diverse ranges of marine invertebrates over a wide geographical area. These microorganisms, however, are phenotypically diverse, and many of the isolates are also resistant to multiple antibiotics. In a previous study, we described a previously unknown association between Vibrio sp. AO1, a luminous bacterium related to the species V. harveyi, and the benthic hydrozoan Aglaophenia octodonta. In this study, we analyzed the susceptibility to antibiotics (ampicillin, streptomycin, tetracycline, or co-trimoxazole = mix of sulfamethoxazole and trimetoprim) of Vibrio sp. AO1 growing in pure culture or in association with its hydroid host by using microcosm experiments. The results of minimum inhibitory concentration (MIC) experiments demonstrated that Vibrio sp. AO1 was highly resistant to ampicillin and streptomycin in pure culture. Nevertheless, these antibiotics, when used at sub-MIC values, significantly reduced the hydroid fluorescence. Co-trimoxazole showed the highest inhibitory effect on fluorescence of A. octodonta. However, in all treatments, the fluorescence was reduced after 48 h, but never disappeared completely around the folds along the hydrocaulus and at the base of the hydrothecae of A. octodonta when the antibiotic was used at concentration completely inhibiting growth in vitro. The apparent discrepancy between the MIC data and the fluorescence patterns may be due to either heterogeneity of the bacterial population in terms of antibiotic susceptibility or specific chemical-physical conditions of the hydroid microenvironment that may decrease the antibiotic susceptibility of the whole population. The latter hypothesis is supported by scanning electron microscope evidence for development of bacterial biofilm on the hydroid surface. On the basis of the results obtained, we infer that A. octodonta might behave as a reservoir of antibiotic multiresistant bacteria

  11. Development of Multiwell-Plate Methods Using Pure Cultures of Methanogens To Identify New Inhibitors for Suppressing Ruminant Methane Emissions.

    Science.gov (United States)

    Weimar, M R; Cheung, J; Dey, D; McSweeney, C; Morrison, M; Kobayashi, Y; Whitman, W B; Carbone, V; Schofield, L R; Ronimus, R S; Cook, G M

    2017-08-01

    Hydrogenotrophic methanogens typically require strictly anaerobic culturing conditions in glass tubes with overpressures of H2 and CO2 that are both time-consuming and costly. To increase the throughput for screening chemical compound libraries, 96-well microtiter plate methods for the growth of a marine (environmental) methanogen Methanococcus maripaludis strain S2 and the rumen methanogen Methanobrevibacter species AbM4 were developed. A number of key parameters (inoculum size, reducing agents for medium preparation, assay duration, inhibitor solvents, and culture volume) were optimized to achieve robust and reproducible growth in a high-throughput microtiter plate format. The method was validated using published methanogen inhibitors and statistically assessed for sensitivity and reproducibility. The Sigma-Aldrich LOPAC library containing 1,280 pharmacologically active compounds and an in-house natural product library (120 compounds) were screened against M. maripaludis as a proof of utility. This screen identified a number of bioactive compounds, and MIC values were confirmed for some of them against M. maripaludis and M. AbM4. The developed method provides a significant increase in throughput for screening compound libraries and can now be used to screen larger compound libraries to discover novel methanogen-specific inhibitors for the mitigation of ruminant methane emissions.IMPORTANCE Methane emissions from ruminants are a significant contributor to global greenhouse gas emissions, and new technologies are required to control emissions in the agriculture technology (agritech) sector. The discovery of small-molecule inhibitors of methanogens using high-throughput phenotypic (growth) screening against compound libraries (synthetic and natural products) is an attractive avenue. However, phenotypic inhibitor screening is currently hindered by our inability to grow methanogens in a high-throughput format. We have developed, optimized, and validated a high

  12. Polycaprolactone fiber meshes provide a 3D environment suitable for cultivation and differentiation of melanocytes from the outer root sheath of hair follicle.

    Science.gov (United States)

    Savkovic, Vuk; Flämig, Franziska; Schneider, Marie; Sülflow, Katharina; Loth, Tina; Lohrenz, Andrea; Hacker, Michael Christian; Schulz-Siegmund, Michaela; Simon, Jan-Christoph

    2016-01-01

    Melanocytes differentiated from the stem cells of human hair follicle outer root sheath (ORS) have the potential for developing non-invasive treatments for skin disorders out of a minimal sample: of hair root. With a robust procedure for melanocyte cultivation from the ORS of human hair follicle at hand, this study focused on the identification of a suitable biocompatible, biodegradable carrier as the next step toward their clinical implementation. Polycaprolactone (PCL) is a known biocompatible material used for a number of medical devices. In this study, we have populated electrospun PCL fiber meshes with normal human epidermal melanocytes (NHEM) as well as with hair-follicle-derived human melanocytes from the outer root sheath (HUMORS) and tested their functionality in vitro. PCL fiber meshes evidently provided a niche for melanocytes and supported their melanotic properties. The cells were tested for gene expression of PAX3, PMEL, TYR and MITF, as well as for proliferation, expression of melanocyte marker proteins tyrosinase and glycoprotein 100 (gp100), L-DOPA-tautomerase enzymatic activity and melanin content. Reduced mitochondrial activity and PAX-3 gene expression indicated that the three-dimensional PCL scaffold supported differentiation rather than proliferation of melanocytes. The monitored melanotic features of both the NHEM and HUMORS cultivated on PCL scaffolds significantly exceeded those of two-dimensional adherent cultures.

  13. Effect of Malytea Scurfpea Fruit on Adhesion and Migration of Melanocyte

    Institute of Scientific and Technical Information of China (English)

    牟宽厚; 冯捷; 张宪旗; 余兵; 周爱民

    2004-01-01

    Objective: To investigate the effect of Malytea Scurfpea fruit (MSF) on adhesion and migra tion of melanocyte. Methods: Human epidermal melanocytes were treated with MSF and examined for its adhesion to bovine serum fibronectin-coated culture plate and for its migration into micropore filters, which werethen compared with those of untreated control. Results: Compared with the control, MSF treated melano cytes were easier to adhere to the dishes and move. Into the filters in a dose-dependent manner, but when the dose of MSF was higher than 150 μg/mi, the adhesion and migration could not be significantly increased. And it was noticed that when the concentrations of MSF was 10μg/ml, significant statistical difference did show between the migration of the treated and untreated melanocytes (P<0.05) though the adhesion was not increased significantly. Conclusion: MSF had the ability to affect melanocyte adhesion and migration. This could explain, in part, the mechanism of MSF in curing vitiligo lesions.

  14. Increased bacterial growth efficiency with environmental variability: results from DOC degradation by bacteria in pure culture experiments

    Directory of Open Access Journals (Sweden)

    M. Eichinger

    2010-06-01

    Full Text Available This paper assesses how considering variation in DOC availability and cell maintenance in bacterial models affects Bacterial Growth Efficiency (BGE estimations. For this purpose, we conducted two biodegradation experiments simultaneously. In experiment one, a given amount of substrate was added to the culture at the start of the experiment whilst in experiment two, the same amount of substrate was added, but using periodic pulses over the time course of the experiment. Three bacterial models, with different levels of complexity, (the Monod, Marr-Pirt and the dynamic energy budget – DEB – models, were used and calibrated using the above experiments. BGE has been estimated using the experimental values obtained from discrete samples and from model generated data. Cell maintenance was derived experimentally, from respiration rate measurements. The results showed that the Monod model did not reproduce the experimental data accurately, whereas the Marr-Pirt and DEB models demonstrated a good level of reproducibility, probably because cell maintenance was built into their formula. Whatever estimation method was used, the BGE value was always higher in experiment two (the periodically pulsed substrate as compared to the initially one-pulsed-substrate experiment. Moreover, BGE values estimated without considering cell maintenance (Monod model and empirical formula were always smaller than BGE values obtained from models taking cell maintenance into account. Since BGE is commonly estimated using constant experimental systems and ignore maintenance, we conclude that these typical methods underestimate BGE values. On a larger scale, and for biogeochemical cycles, this would lead to the conclusion that, for a given DOC supply rate and a given DOC consumption rate, these BGE estimation methods overestimate the role of bacterioplankton as CO2 producers.

  15. Increased bacterial growth efficiency with environmental variability: results from DOC degradation by bacteria in pure culture experiments

    Directory of Open Access Journals (Sweden)

    M. Eichinger

    2010-02-01

    Full Text Available This paper assesses how considering variation in DOC availability and cell maintenance in bacterial models affects Bacterial Growth Efficiency (BGE estimations. For this purpose, we conducted two biodegradation experiments simultaneously. In experiment one, a given amount of substrate was added to the culture at the start of the experiment whilst in experiment two, the same amount of substrate was added, but using periodic pulses over the time course of the experiment. Three bacterial models, with different levels of complexity, (the Monod, Marr-Pirt and the dynamic energy budget (DEB model, were used, and calibrated using the above experiments. BGE has been estimated using the experimental values obtained from discrete samples and from model generated data. Cell maintenance was derived experimentally, from respiration rate measurements. The results showed that the Monod model did not reproduce the experimental data accurately, whereas the Marr-Pirt and DEB models demonstrated a good level of reproducibility, probably because cell maintenance was built into their formula. Whatever estimation method was used, the BGE value was always higher in experiment two (the periodically pulsed substrate as compared to the initially one-pulsed-substrate experiment. Moreover, BGE values estimated without considering cell maintenance (Monod model and empirical formula were always smaller than BGE values obtained from models taking cell maintenance into account. Since BGE is commonly estimated using constant experimental systems and ignore maintenance, we conclude that these typical methods underestimate BGE values. On a larger scale, and for biogeochemical cycles, this would lead to the conclusion that, for a given DOC supply rate and a given DOC consumption rate, these BGE estimation methods overestimate the role of bacterioplankton as CO2 producers.

  16. Remote detection of laser-induced autofluorescence on pure cultures of fungal and bacterial strains and their analysis with multivariate techniques

    Science.gov (United States)

    Raimondi, Valentina; Palombi, Lorenzo; Cecchi, Giovanna; Lognoli, David; Trambusti, Massimo; Gomoiu, Ioana

    2007-05-01

    Remotely sensed laser-induced autofluorescence spectra of pure cultures of fungal strains ( Aureobasidium pullulans, Verticillium sp.) and of bacterial strains ( Bacillus sp., Pseudomonas sp.) are presented. The strains were isolated from samples collected in a Roman archaeological site ( Tropaeum Traiani) near Constanta, Romania. The fluorescence spectra were detected in vivo from a distance of 25 m in the outdoor, using a high spectral resolution fluorescence LIDAR featuring a UV laser (XeCl@308 nm) as an excitation source. All the examined strains, except for the A. pullulans, showed fluorescence features such to allow their characterisation by processing data with multivariate techniques. Both Principal Component Analysis and Cluster Analysis were applied to the data set and compared to discriminate between the examined strains. Results demonstrate the feasibility of fluorescence-based detection and characterisation of fungi and bacteria in the outdoor with a high spectral resolution fluorescence LIDAR. In addition, they show that the proposed processing methods offer a means to discriminate between the fluorescence features due to the investigated samples and that of a fluorescence background of a known spectral shape, as that of the culture medium. This can be exploited for the remote fluorescence mapping of heterotrophic organisms on stone surfaces when the latter show a typical broad fluorescence band.

  17. Dynamic model-based analysis of furfural and HMF detoxification by pure and mixed batch cultures of S. cerevisiae and S. stipitis.

    Science.gov (United States)

    Hanly, Timothy J; Henson, Michael A

    2014-02-01

    Inhibitory compounds that result from biomass hydrolysis are an obstacle to the efficient production of second-generation biofuels. Fermentative microorganisms can reduce compounds such as furfural and 5-hydroxymethyl furfural (HMF), but detoxification is accompanied by reduced growth rates and ethanol yields. In this study, we assess the effects of these furan aldehydes on pure and mixed yeast cultures consisting of a respiratory deficient mutant of Saccharomyces cerevisiae and wild-type Scheffersomyces stipitis using dynamic flux balance analysis. Uptake kinetics and stoichiometric equations for the intracellular reduction reactions associated with each inhibitor were added to genome-scale metabolic reconstructions of the two yeasts. Further modification of the S. cerevisiae metabolic network was necessary to satisfactorily predict the amount of acetate synthesized during HMF reduction. Inhibitory terms that captured the adverse effects of the furan aldehydes and their corresponding alcohols on cell growth and ethanol production were added to attain qualitative agreement with batch experiments conducted for model development and validation. When the two yeasts were co-cultured in the presence of the furan aldehydes, inoculums that reduced the synthesis of highly toxic acetate produced by S. cerevisiae yielded the highest ethanol productivities. The model described here can be used to generate optimal fermentation strategies for the simultaneous detoxification and fermentation of lignocellulosic hydrolysates by S. cerevisiae and/or S. stipitis.

  18. Model of human epidermis reconstructed in vitro with keratinocytes and melanocytes on dead de-epidermized human dermis

    Directory of Open Access Journals (Sweden)

    Jussara Rehder

    Full Text Available CONTEXT: Recent progress in the field of epithelial culture techniques has allowed the development of culture systems in which the reconstructed epidermis presents characteristics of morphological differentiation similar to those seen in vivo. Human epidermis reconstructed in vitro may be used as the best alternative for the in vitro testing of the toxicology and efficiency of products for topical use, as well as in the treatment of skin burns and chronic skin ulcers. OBJECTIVE: To demonstrate a method for obtaining human epidermis reconstructed in vitro, using keratinocytes and melanocytes cultivated on dead de-epidermized human dermis. TYPE OF STUDY: Experimental/laboratory. SETTING: Skin Cell Culture Laboratory of the Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil. PROCEDURE: Human keratinocytes and melanocytes cultured in vitro were grown on a biological matrix (dead de-epidermized human dermis and the system was kept at an air-liquid interface, in a suitable culturing medium, until a stratified human epidermis was formed, maintaining the histological characteristics of the epidermis in vivo. RESULTS: It was histologically demonstrated that it is possible to reproduce a differentiated epidermis through keratinocytes and melanocytes cultured on dead de-epidermized human dermis, thus obtaining a correctly positioned human epidermis reconstructed in vitro with functional keratinocytes and melanocytes that is similar to in vivo epidermis. CONCLUSIONS: It is possible to obtain a completely differentiated human epidermis reconstructed in vitro from keratinocyte and melanocyte cultures on a dead de-epidermized human dermis.

  19. Congenital melanocytic nevi: Catch them early!

    Directory of Open Access Journals (Sweden)

    Karthika Natarajan

    2013-01-01

    Full Text Available We report a 2-week-old neonate with a large congenital melanocytic nevus over face treated with surgical curettage followed by a combination of carbon dioxide laser and Q-switched neodymium-doped yttrium aluminum garnet lasers. The results were satisfactory with near complete resolution after 1 year of age. This case is reported to emphasize the timely management and the importance of curettage prior to development of rete ridges.

  20. Possible Existence of Melanocytes or Melanoblasts in Human Sebaceous Glands

    OpenAIRE

    Jang, Yong Hyun; Kim, Sang Lim; Lee, Jin Sub; Kwon, Kun-Young; Lee, Seok-Jong; Kim, Do Won; Lee, Weon Ju

    2014-01-01

    Background Melanocytes are present in both basal epidermis and hair follicles. Melanocyte stem cells have been found in hair follicle bulge. During embryogenesis, the outer cells of the bulge differentiate into the sebaceous gland (SG) and proliferate. Objective To identify and determine the distribution and morphological characteristics of melanocytes in human SGs. Methods A total of 171 biopsy specimens of face and scalp were studied. Of these specimens, 103 samples contained SGs. We conduc...

  1. Limbal melanocytes support limbal epithelial stem cells in 2D and 3D microenvironments.

    Science.gov (United States)

    Dziasko, Marc A; Tuft, Stephen J; Daniels, Julie T

    2015-09-01

    Human limbal epithelial stem cells (LESCs) are essential for the maintenance of the corneal epithelium of the ocular surface. LESCs are located within limbal crypts between the palisades of Vogt in the limbus; the interface between the peripheral cornea and conjunctiva. The limbal crypts have been proposed as a LESC niche owing to their support of epithelial cells, which can form holoclone colonies in vitro. Closely associated with the limbal crypts is a concentrated population of melanocytes. The anatomical location and close proximity to putative LESC suggests that melanocytes might play a role in maintenance of these stem cells in the niche. The aim of this study was to assess the ability of human limbal melanocytes (hLM) to support the expansion of human limbal epithelial cells (LECs) in vitro as an indicator of functional cell-cell interaction. After observing that hLM co-localize with clusters of compact epithelial cells in the native limbal crypts, hLM were isolated from crypt-rich cadaveric limbal biopsies and used as feeders for the culture of LECs. Interestingly, LECs grown on mitotically active hLM were able to generate large epithelial colonies that contained small and compact cells with morphological stem cell characteristics. Immunocytochemistry revealed that LECs expanded on hLM were positive for the expression of the putative stem cell markers CK15, Bmi-1 and p63α and negative for the marker of terminal cell differentiation CK3. LECs and hLM were finally co-cultured on RAFT (real architecture for 3D tissue) collagen tissue equivalents. In 3D co-cultures, hLM promoted multi-layering of the epithelial sheet in which basal cells were maintained in an undifferentiated state. Taken together, these observations suggest melanocytes could play an important role in the maintenance of LESCs in the native human limbal stem cell niche.

  2. Enumeration of total bacteria and bacteria with genes for proteolytic activity in pure cultures and in environmental samples by quantitative PCR mediated amplification.

    Science.gov (United States)

    Bach, H-J; Tomanova, J; Schloter, M; Munch, J C

    2002-05-01

    Real-time quantitative PCR assays were developed for the absolute quantification of different groups of bacteria in pure cultures and in environmental samples. 16S rRNA genes were used as markers for eubacteria, and genes for extracellular peptidases were used as markers for potentially proteolytic bacteria. For the designed 16S rDNA TaqMan assay, specificity of the designed primer-probe combination for eubacteria, a high amplification efficiency over a wide range of starting copy numbers and a high reproducibility is demonstrated. Cell concentrations of Bacillus cereus, B. subtilis and Pseudomonas fluorescens in liquid culture were monitored by TaqMan-PCR using the 16S rDNA target sequence of Escherichia coli as external standard for quantification. Results agree with plate counts and microscopic counts of DAPI stained cells. The significance of 16S rRNA operon multiplicity to the quantification of bacteria is discussed.Furthermore, three sets of primer pair together with probe previously designed for targeting different classes of bacterial extracellular peptidases were tested for their suitability for TaqMan-PCR based quantification of proteolytic bacteria. Since high degeneracy of the probes did not allow accurate quantification, SybrGreen was used instead of molecular probes to visualize and quantify PCR products during PCR. The correlation between fluorescence and starting copy number was of the same high quality as for the 16S rDNA TaqMan assay for all the three peptidase gene classes. The detected amount of genes for neutral metallopeptidase of B. cereus, for subtilisin of B. subtilis and for alkaline metallopeptidase of P. fluorescens corresponded exactly to the numbers of bacteria investigated by the 16S rDNA targeting assay. The developed assays were applied for the quantification of bacteria in soil samples.

  3. Optimization of a high-throughput CTAB-based protocol for the extraction of qPCR-grade DNA from rumen fluid, plant and bacterial pure cultures.

    Science.gov (United States)

    Minas, Konstantinos; McEwan, Neil R; Newbold, Charles Jamie; Scott, Karen P

    2011-12-01

    The quality and yield of extracted DNA are critical for the majority of downstream applications in molecular biology. Moreover, molecular techniques such as quantitative real-time PCR (qPCR) are becoming increasingly widespread; thus, validation and cross-laboratory comparison of data require standardization of upstream experimental procedures. DNA extraction methods depend on the type and size of starting material(s) used. As such, the extraction of template DNA is arguably the most significant variable when cross-comparing data from different laboratories. Here, we describe a reliable, inexpensive and rapid method of DNA purification that is equally applicable to small or large scale or high-throughput purification of DNA. The protocol relies on a CTAB-based buffer for cell lysis and further purification of DNA with phenol : chloroform : isoamyl alcohol. The protocol has been used successfully for DNA purification from rumen fluid and plant cells. Moreover, after slight alterations, the same protocol was used for large-scale extraction of DNA from pure cultures of Gram-positive and Gram-negative bacteria. The yield of the DNA obtained with this method exceeded that from the same samples using commercial kits, and the quality was confirmed by successful qPCR applications.

  4. Interferon-γ induces senescence in normal human melanocytes.

    Directory of Open Access Journals (Sweden)

    Suiquan Wang

    Full Text Available BACKGROUND: Interferon-γ (IFN-γ plays an important role in the proceedings of vitiligo through recruiting lymphocytes to the lesional skin. However, the potential effects of IFN-γ on skin melanocytes and the subsequent contribution to the vitiligo pathogenesis are still unclear. OBJECTIVE: To investigate the effects of IFN-γ on viability and cellular functions of melanocytes. METHODS: Primary human melanocytes were treated with IFN-γ. Cell viability, apoptosis, cell cycle melanin content and intracellular reactive oxygen species (ROS level were measured. mRNA expression was examined by real-time PCR. The release of interleukin 6 (IL-6 and heat shock protein 70 (HSP-70 was monitored by ELISA. β-galactosidase staining was utilized to evaluate melanocyte senescence. RESULTS: Persistent IFN-γ treatment induced viability loss, apoptosis, cell cycle arrest and senescence in melanocytes. Melanocyte senescence was characterized as the changes in pigmentation and morphology, as well as the increase of β-galactosidase activity. Increase of p21Cip1/Waf1 protein was evident in melanocytes after IFN-γ treatment. IFN-γ induction of senescence was attenuated by siRNAs against p21, Janus kinase 2 (JAK2 or signal transducer and activator of transcription 1 (STAT1, but not by JAK1 siRNA nor by p53 inhibitor pifithrin-α. IFN-γ treatment increased the accumulation of intracellular ROS in melanocytes, while ROS scavenger N-acetyl cysteine (NAC effectively inhibited IFN-γ induced p21 expression and melanocyte senescence. IL-6 and HSP-70 release was significantly induced by IFN-γ treatment, which was largely inhibited by NAC. The increase of IL-6 and HSP-70 release could also be observed in senescent melanocytes. CONCLUSION: IFN-γ can induce senescence in melanocytes and consequently enhance their immuno-competency, leading to a vitiligo-prone milieu.

  5. Induction of Chemokine Secretion and Monocyte Migration by Human Choroidal Melanocytes in Response to Proinflammatory Cytokines

    DEFF Research Database (Denmark)

    Jehs, Tina; Faber, Carsten; Udsen, Maja S.

    2016-01-01

    Purpose: To determine to which extent inflammatory cytokines affect chemokine secretion by primary human choroidal melanocytes (HCMs), their capacity to attract monocytes, and whether HCMs are able to influence the proliferation of activated T cells. Methods: Primary cultures of HCMs were...... established from eyes of 13 donors. Human choroidal melanocytes were stimulated with IFN-γ and TNF-α or with supernatant from activated T cells (T-cell–conditioned media [TCM]). Gene expression analysis was performed by using microarrays. Protein levels were quantified with ELISA or cytometric bead array....... Supernatants of HCMs were assessed for the capability to attract monocytes in a transwell plate. Proliferation of activated T cells was assessed in a direct coculture with HCMs by a [3H]-thymidine incorporation assay. Results: Stimulation of HCMs with TCM or IFN-γ and TNF-α resulted in increased expression...

  6. Cytotoxic T lymphocyte responses against melanocytes and melanoma

    Directory of Open Access Journals (Sweden)

    Schwartz Erich J

    2011-07-01

    Full Text Available Abstract Background Vitiligo is a common toxicity associated with immunotherapy for melanoma. Cytotoxic T lymphocytes (CTLs against melanoma commonly target melanoma-associated antigens (MAAs which are also expressed by melanocytes. To uncouple vitiligo from melanoma destruction, it is important to understand if CTLs can respond against melanoma and melanocytes at different levels. Methods To understand the dichotomous role of MAA-specific CTL, we characterized the functional reactivities of established CTL clones directed to MAAs against melanoma and melanocyte cell lines. Results CTL clones generated from melanoma patients were capable of eliciting MHC-restricted, MAA-specific lysis against melanocyte cell lines as well as melanoma cells. Among the tested HLA-A*0201-restricted CTL clones, melanocytes evoked equal to slightly higher degranulation and cytolytic responses as compared to melanoma cells. Moreover, MAA-specific T cells from vaccinated patients responded directly ex vivo to melanoma and melanocytes. Melanoma cells express slightly higher levels of MART-1 and gp100 than melanocytes as measured by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR and immunohistochemistry. Conclusions Our data suggest that CTLs respond to melanoma and melanocytes equally in vitro and directly ex vivo.

  7. Neurotized congenital melanocytic nevus resembling a pigmented neurofibroma

    Directory of Open Access Journals (Sweden)

    Nidhi Singh

    2015-01-01

    Full Text Available Neurotized congenital melanocytic nevus and pigmented neurofibroma (PNF are close mimics and pose a clinicopathological challenge. We present a case of pigmented hypertrichotic plaque over lumbosacral region and discuss the differential diagnosis and its clinical, histopathological and immunohistochemistry features which may aid in differentiation. We highlight the difficulties faced in differentiating neurotized congenital melanocytic nevus from pigmented neurofibroma.

  8. Transport of melanocytes by chitosan membrane: an animal experiment%壳聚糖膜转载黑素细胞动物实验研究

    Institute of Scientific and Technical Information of China (English)

    董东; 许思原; 傅立芳; 王涛; 许爱娥

    2011-01-01

    Objective To study the feasibility of using chitosan membrane to carry and transport melanocytes, in order to refine the technique for melanocyte transplantation with chitosan membrane. Methods Melanocytes were inoculated onto chitosan membrane and cultured for a period of time, then, electron microscopy,MTT assay and NaOH assay were carried out to estimate the adherence, growth and melanogenesis of the melanocytes. Skin wound surface was prepared in 12 nude mice, which were equally divided into 3 groups, test group inoculated with melanocytes on chitosan membrane, negative control group I treated with chitosan membrane without melanocytes, and negative control group II directly dressed immediately after the preparation of wound surface. On day 10 and 20 after the transplantation, confocal laser microscopy and immunohistochemistry were performed to observe the migration of melanocytes into the skin wound surface. Results Scanning electron microscopy and inverted microscopy showed that melanocytes were evenly distributed on and adhered well to the underlying chitosan membrane. As the growth curve of melanocytes demonstrated, chitosan membrane could support the normal growth of melanocytes, and no significant difference was observed in the synthesized melanin content between melanocytes cultured on the chitosan membrane and those in culture disks (0.087 ± 0.027 vs. 0.101 ± 0.036, t = 0.79, P > 0.05). Melanocytes were seen at the transplantation sites by confocal laser microscopy, and biopsy specimens from the transplantation sites stained positive for antimelan-A monoclonal antibody. Conclusions Melanocytes can adhere to and grow on the chitosan membrane,which can facilitate the migration of melanocytes to the transplantation sites in animals with the maintenance of biological activity of melanocytes.%目的 探讨壳聚糖膜搭载、转运黑素细胞的可行性,探索利用壳聚糖膜改进黑素细胞移植技术.方法 使用电镜、MTT法、NaOH裂解法

  9. New Insights into the Molecular Distinction of Dysplastic Nevi and Common Melanocytic Nevi-Highlighting the Keratinocyte-Melanocyte Relationship.

    Science.gov (United States)

    Eliades, Philip; Tsao, Hensin

    2016-10-01

    Mitsui et al. approach the problem of differentiating dysplastic nevi from common melanocytic nevi through a molecular lens. Whereas most of the literature on this topic shines the spotlight toward melanocytes, the focus of this paper is shifted to the tumor microenvironment. Using microarrays, reverse transcriptase-PCR, and immunohistochemistry, their results emphasize the role of keratinocyte dysplasia within dysplastic nevi.

  10. Melanocyte Transformation Associated with Substrate Adhesion Impediment

    Directory of Open Access Journals (Sweden)

    Sueli M. Oba-Shinjo

    2006-03-01

    Full Text Available Exclude experimental models of malignant transformation employ chemical and physical carcinogens or genetic manipulations to study tumor progression. In this work, different melanoma cell lines were established after submitting a nontumorigenic melanocyte lineage (melan-a to sequential cycles of forced anchorage impediment. The great majority of these cells underwent anoikis when maintained in suspension. After one deadhesion cycle, phenotypic alterations were noticeable in the few surviving cells, which became more numerous and showed progressive alterations after each adhesion impediment step. No significant differences in cell surface expression of integrins were detected, but a clear electrophoretic migration shift, compatible with an altered glycosylation pattern, was observed for β1 chain in transformed cell lines. In parallel, a progressive enrichment of tri- and tetra-antennary N-glycans was apparent, suggesting increased N-acetylglucosaminyl-transferase V activity. Alterations both in proteoglycan glycosylation pattern and core protein expression were detected during the transformation process. In conclusion, this model corroborates the role of adhesion state as a promoting agent in transformation process and demonstrates that cell adhesion disturbances may act as carcinogenic stimuli, at least for a nontumorigenic immortalized melanocyte lineage. These findings have intriguing implications for in vivo carcinogenesis, suggesting that anchorage independence may precede, and contribute to, neoplastic conversion.

  11. Primary Cilia Negatively Regulate Melanogenesis in Melanocytes and Pigmentation in a Human Skin Model

    Science.gov (United States)

    Kim, Eun Sung; Park, So Jung; Bae, Il-Hong; Jo, Yoon Kyung; Jeong, In Young; Kim, Hyoung-June; Lee, Youngjin; Park, Hea Chul; Jeon, Hong Bae; Kim, Ki Woo; Lee, Tae Ryong; Cho, Dong-Hyung

    2016-01-01

    The primary cilium is an organelle protruding from the cell body that senses external stimuli including chemical, mechanical, light, osmotic, fluid flow, and gravitational signals. Skin is always exposed to the external environment and responds to external stimuli. Therefore, it is possible that primary cilia have an important role in skin. Ciliogenesis was reported to be involved in developmental processes in skin, such as keratinocyte differentiation and hair formation. However, the relation between skin pigmentation and primary cilia is largely unknown. Here, we observed that increased melanogenesis in melanocytes treated with a melanogenic inducer was inhibited by a ciliogenesis inducer, cytochalasin D, and serum-free culture. However, these inhibitory effects disappeared in GLI2 knockdown cells. In addition, activation of sonic hedgehog (SHH)-smoothened (Smo) signaling pathway by a Smo agonist, SAG inhibited melanin synthesis in melanocytes and pigmentation in a human skin model. On the contrary, an inhibitor of primary cilium formation, ciliobrevin A1, activated melanogenesis in melanocytes. These results suggest that skin pigmentation may be regulated partly by the induction of ciliogenesis through Smo-GLI2 signaling. PMID:27941997

  12. Effective intra-S checkpoint responses to UVC in primary human melanocytes and melanoma cell lines.

    Science.gov (United States)

    Cordeiro-Stone, Marila; McNulty, John J; Sproul, Christopher D; Chastain, Paul D; Gibbs-Flournoy, Eugene; Zhou, Yingchun; Carson, Craig; Rao, Shangbang; Mitchell, David L; Simpson, Dennis A; Thomas, Nancy E; Ibrahim, Joseph G; Kaufmann, William K

    2016-01-01

    The objective of this study was to assess potential functional attenuation or inactivation of the intra-S checkpoint during melanoma development. Proliferating cultures of skin melanocytes, fibroblasts, and melanoma cell lines were exposed to increasing fluences of UVC and intra-S checkpoint responses were quantified. Melanocytes displayed stereotypic intra-S checkpoint responses to UVC qualitatively and quantitatively equivalent to those previously demonstrated in skin fibroblasts. In comparison with fibroblasts, primary melanocytes displayed reduced UVC-induced inhibition of DNA strand growth and enhanced degradation of p21Waf1 after UVC, suggestive of enhanced bypass of UVC-induced DNA photoproducts. All nine melanoma cell lines examined, including those with activating mutations in BRAF or NRAS oncogenes, also displayed proficiency in activation of the intra-S checkpoint in response to UVC irradiation. The results indicate that bypass of oncogene-induced senescence during melanoma development was not associated with inactivation of the intra-S checkpoint response to UVC-induced DNA replication stress.

  13. Defects in ErbB-dependent establishment of adult melanocyte stem cells reveal independent origins for embryonic and regeneration melanocytes.

    Directory of Open Access Journals (Sweden)

    Keith A Hultman

    2009-07-01

    Full Text Available Adult stem cells are responsible for maintaining and repairing tissues during the life of an organism. Tissue repair in humans, however, is limited compared to the regenerative capabilities of other vertebrates, such as the zebrafish (Danio rerio. An understanding of stem cell mechanisms, such as how they are established, their self-renewal properties, and their recruitment to produce new cells is therefore important for the application of regenerative medicine. We use larval melanocyte regeneration following treatment with the melanocytotoxic drug MoTP to investigate these mechanisms in Melanocyte Stem Cell (MSC regulation. In this paper, we show that the receptor tyrosine kinase, erbb3b, is required for establishing the adult MSC responsible for regenerating the larval melanocyte population. Both the erbb3b mutant and wild-type fish treated with the ErbB inhibitor, AG1478, develop normal embryonic melanocytes but fail to regenerate melanocytes after MoTP-induced melanocyte ablation. By administering AG1478 at different time points, we show that ErbB signaling is only required for regeneration prior to MoTP treatment and before 48 hours of development, consistent with a role in establishing MSCs. We then show that overexpression of kitla, the Kit ligand, in transgenic larvae leads to recruitment of MSCs, resulting in overproliferation of melanocytes. Furthermore, kitla overexpression can rescue AG1478-blocked regeneration, suggesting that ErbB signaling is required to promote the progression and specification of the MSC from a pre-MSC state. This study provides evidence that ErbB signaling is required for the establishment of adult MSCs during embryonic development. That this requirement is not shared with the embryonic melanocytes suggests that embryonic melanocytes develop directly, without proceeding through the ErbB-dependent MSC. Moreover, the shared requirement of larval melanocyte regeneration and metamorphic melanocytes that develops at

  14. Matrigel supports neural, melanocytic and chondrogenic differentiation of trunk neural crest cells.

    Science.gov (United States)

    Ramos-Hryb, Ana B; Da-Costa, Meline C; Trentin, Andréa G; Calloni, Giordano W

    2013-01-01

    The neural crest (NC) is composed of highly multipotent precursor cells able to differentiate into both neural and mesenchymal phenotypes. Until now, most studies focusing on NC cell differentiation have been performed with traditional two-dimensional (2D) cell culture systems. However, such culture systems do not reflect the complex three-dimensional (3D) microenvironments of in vivo NC cells. To address this limitation, we have developed a method of Matrigel™ coating to create 2D and 3D microenvironments in the same culture well. When we performed cultures of trunk neural crest cells (TNCCs) on three different lots of basement membrane matrix (Matrigel™), we observed that all analyzed Matrigel™ lots were equally efficient in allowing the appearance of glial cells, neurons, melanocytes, smooth muscle cells and chondrocytes. We further observed that chondrocytes were found predominantly in the 3D microenvironment, whereas smooth muscle cells were almost exclusively located in the 2D microenvironment. Glial cells were present in both environments, but with broader quantities on the 2D surface. Melanocytes and neurons were equally distributed in both 2D and 3D microenvironments, but with distinct morphologies. It is worth noting the higher frequency of chondrocytes detected in this study using the 3D Matrigel™ microenvironment compared to previous reports of chondrogenesis obtained from TNCCs on traditional 2D cultures. In conclusion, Matrigel™ represents an attractive scaffold to study NC multipotentiality and differentiation, since it permits the appearance of the major NC phenotypes.

  15. Multiple roles of NF1 in the melanocyte lineage.

    Science.gov (United States)

    Larribère, Lionel; Utikal, Jochen

    2016-07-01

    NF1 is a tumour suppressor gene, germline mutations of which lead to neurofibromatosis type 1 syndrome. Patients develop benign tumours from several types of cells including neural crest-derived cells. NF1 somatic mutations also occur in 15% of sporadic melanoma, a cancer originating from melanocytes. Evidence now suggests the involvement of NF1 mutations in melanoma resistance to targeted therapies. Although NF1 is ubiquitously expressed, genetic links between NF1 and genes involved in melanocyte biology have been described, implying the lineage-specific mechanisms. In this review, we summarize and discuss the latest advances related to the roles of NF1 in melanocyte biology and in cutaneous melanoma.

  16. Comparative Study of Efficacy of Epidermal Melanocyte Transfer Versus Hair Follicular Melanocyte Transfer in Stable Vitiligo

    Science.gov (United States)

    Donaparthi, Navya; Chopra, Ajay

    2016-01-01

    Background: Vitiligo surgery has come up a long way from punch skin grafts to epidermal cell suspension and latest to the extracted hair follicle outer root sheath cell suspension (EHFORSCS) transplantation. The progressive development from one technique to the other is always on a quest for the best. In the latest development, EHFORSCS, which is an enriched source of follicular inactive melanocyte (melanocyte stem cells), seems to be a good addition to the prevailing cell-based therapies for vitiligo. However, it needs to be explored further in larger, clinical trials. Methodology: A total of 11 patients with sixty stable vitiligo sites attending dermatology outpatient department were included for the open-labeled, prospective, comparative study. The sites were sequentially distributed into two groups of thirty each. Sites of one group were subjected to epidermal melanocyte transfer (EMT) and the others to hair follicular melanocyte transfer (HFMT). Response to treatment was evaluated on the basis of degree of repigmentation; final evaluation of area of involvement was done after completion of 6 months. Results: At the end of 6 months, repigmentation >90% was observed in 83.33% patches of EMT group and 43.33% in HFMT group. Repigmentation >75% was observed in 90% of patches in Group A and 43.34% of patches in Group B, respectively. There was statistically significant difference in the overall pigmentation between these two groups. Conclusion: Both noncultured autologous epidermal cell suspension transfer and noncultured EHFORSCS transfer are safe and effective surgical modalities in the management of stable vitiligo though EMT has shown a better response in the present study. Outer root sheath cell suspension transfer is a novel, minimally invasive technique in its nascent stage in the surgical management of vitiligo which requires further larger clinical trials for evaluation of its efficacy. PMID:27904182

  17. Primary transpupillary thermotherapy of choroidal melanocytic lesions

    Directory of Open Access Journals (Sweden)

    Kaan Gündüz

    2011-01-01

    Full Text Available Purpose : To evaluate the role of primary transpupillary thermotherapy (TTT in the treatment of choroidal melanocytic lesions. Materials and Methods : Retrospective chart review of 24 patients (24 eyes with choroidal melanocytic lesions, including 20 choroidal melanoma and four choroidal nevus treated with primary TTT. Choroidal nevus cases treated with primary TTT either demonstrated risk factors for growth into an early melanoma or had overlying choroidal neovascularization. Results : The mean initial tumor basal diameter was 6.6 (3.0-10.0 mm and the mean initial tumor thickness was 3.0 (1.0-5.0 mm. The mean number of TTT sessions was 2.5 (1-6. The mean decrease in tumor thickness was 1.2 mm (from 3.0 to 1.8 mm at a mean follow-up of 22.7 (range 3-90 months. On the LogMar scale, visual acuity was stable at 1.0. Complications occurred in 50% of eyes. The most frequent complications were vitreous hemorrhage [5 patients (20.8%], focal cataract [5 patients (20.8%], iris atrophy [4 patients (16.6%] and posterior synechia [4 patients (16.6%]. There was no significant difference in the complication rate with respect to tumor thickness >3 mm versus tumor thickness ≤3 mm and juxtapapillary versus nonjuxtapapillary location (Fisher′s exact test, P>0.05. Kaplan-Meier curves showed that 9% of eyes develop recurrence by 1 year and 27% develop recurrence by 5 years after primary TTT. Two eyes (8.3% were enucleated because of neovascular glaucoma and one eye (4.1% was exenterated because of extraocular tumor recurrence. Globe salvage was achieved in 21 patients (87.5%. One patient (4.1% with extraocular tumor recurrence developed liver metastasis and expired. Conclusions : Although TTT may be useful in the treatment of small choroidal melanocytic lesions, the high complication and recurrence rates warrant close monitoring of patients after primary TTT even when a flat chorioretinal scar has been achieved.

  18. A comparative study of the effects of different low-level lasers on the proliferation, viability, and migration of human melanocytes in vitro.

    Science.gov (United States)

    AlGhamdi, Khalid M; Kumar, Ashok; Ashour, Abdelkader E; AlGhamdi, Attieh A

    2015-07-01

    The aim of this study was to investigate the effects of different low-level laser therapies (LLLTs) of various wavelengths and energies on normal cultured human melanocytes. Various studies have shown the effects of LLLs on various types of cultured cells. Presently, little is known about the biological effects of LLLTs on melanocytes. Melanocytes were exposed to LLLT at 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, or 5.0 J/cm(2) using a blue (457 nm), red (635 nm), or ultraviolet (UV) (355 nm) laser. Melanocyte viability, proliferation, and migration were monitored at 72 h after irradiation. The blue (P migration at 0.5 to 4.0 J/cm(2) (P migration at 0.5 to 1.0 J/cm(2) and 0.5 to 2.0 J/cm(2), respectively, compared with controls. LLLT at low energy densities is able to significantly increase melanocyte viability, proliferation, and migration in vitro, and at higher energy densities, it gives non-stimulatory results. Additionally, the blue laser was the best among the three lasers. These findings might have potential application in vitiligo treatment in future.

  19. Melanocyte response to gravitational stress: an overview with a focus on the role of cyclic nucleotides

    Science.gov (United States)

    Ivanova, Krassimira; Tsiockas, Wasiliki; Eiermann, Peter; Hauslage, Jens; Hemmersbach, Ruth; Block, Ingrid; Gerzer, Rupert

    Human melanocytes are responsible for skin pigmentation by synthesizing the pigment melanin. A well known modulator of melanogenesis is the second messenger adenosine 3',5'-cyclic monophos-phate (cAMP). It has also been reported that the nitric oxide (NO)/soluble guanylyl cyclase (sGC)/guanosine 3',5'-cyclic monophosphate (cGMP) pathway is involved in UVB-induced melanogenesis. Melanin acts as a scavenger for free radicals during oxidative stress, but it may additionally act as a photosensitizer that generates active oxygen species upon UV radiation, which may initiate hypopigmentary disorders (e.g., vitiligo) as well as UV-induced oncogene cell transformation. Melanoma, a deadly skin cancer which arises from transformed melanocytes, is characterized by a resistance to chemotherapy. In our studies we were able to show that hu-man melanocytic cells differentially respond to gravitational stress. Hypergravity (up to 5 g for 24 h) stimulated cGMP efflux in cultured human melanocytes and non-metastatic melanoma cells, but not in metastatic phenotypes under the conditions of limited degradation [e.g., in the presence of phosphodiesterase (PDE) inhibitors] or stimulated synthesis of cGMP [e.g., by NO donors, but not natriuretic peptides], whereas cellular proliferation and morphology were not altered. Interestingly, long-term exposure to hypergravity stimulated an increase in both intra-cellular as well as extracellular cAMP levels as well as melanogenesis in pigmented melanocytes and non-metastatic melanoma cells. As some cAMP-PDEs are regulated by cGMP, it seems that the hypergravity-induced alteration of melanocyte pigmentation could be a result of a cross-talk between these two cyclic nucleotides. Hypergravity induced further an increase in the mRNA and protein levels of the selective cGMP and cAMP exporters, the multidrug resistance proteins (MRP) 4 and 5 -but not 8 -, whereas simulated microgravity (up to 1.21x10-2 g for 24 h) -provided by a fast-rotating clinostat

  20. 多巴色素异构酶突变对体外培养黑素小体成熟和抗氧化应激能力的影响%Effects of mutation in dopachrome tautomerase on melanosome maturation and anti-oxidative potential in cultured melanocytes

    Institute of Scientific and Technical Information of China (English)

    万静; 刘小明; 雷铁池; 徐世正

    2009-01-01

    Objective To investigate whether the mutation in dopachrome tautomerase (Dct) affects melanosome maturation and anti-oxidative potential in cultured melanocytes (MCs). Methods Slaty and melan-a MCs were derived from the skins of neonatal DctSlt and C57 BI/6J mice respectively. Their detailed melanosome structures were examined with a transmission electron microscopy (TEM) and their eu-melanin granules characterized by Fontana-Masson staining. Furthermore, the tyrosinase activity and three melanogenic proteins, I.e. , tyrosinase, tyrosinase-related protein 1 and Dct, were also measured with a spectrophotometery method or Western blot assay. The level of intraceilular reactive oxygen species (ROS) was monitored by 2, 7-dichlorofluorescin diacetate (DCF-DA) labeling. Results Mature stage Ⅳ melanosomes markedly decreased in slaty MCs under TEM. The brownish granules stained with Fontana-Masson silver method were far less in slaty MCs than in melan-a MCs. The cell pellet of slaty MCs was white in color, but the similarities between slaty and melan-a were found in tyrosinase activity and its protein expression. The relative intensity of DCF fluorescence was 8.9±0.7 for slaty melanocytes versus 8.9±2.5 for melan-a melanocytes prior to UVA irradiation, but an abrupt ROS production was merely observed in slaty MCs (18.0±0.3) other than in melan-a MCs (13.6±0.3) after UVA exposure. There was statistical difference between these two cell lines in ROS level upon UVA irradiation (P=0.024). Conclusion The mutation in Det causes hypo-pigmented phenotype in cultured slaty MCs, inhibits melanosome maturation and decreases anti-oxidative capacity especially in the presence of UVA-induced oxidative stress.%目的 研究多巴色素异构酶(Dct)突变对黑素细胞黑素小体成熟和活性氧基(ROS)清除能力的影响.方法 用透射电子显微镜技术和Fontana-Masson嗜银染色观察小鼠Dct突变slaty黑素细胞和野生型melan-a黑素细胞胞内黑素小体

  1. Role of Melanin in Melanocyte Dysregulation of Reactive Oxygen Species

    Directory of Open Access Journals (Sweden)

    Noah C. Jenkins

    2013-01-01

    Full Text Available We have recently reported a potential alternative tumor suppressor function for p16 relating to its capacity to regulate oxidative stress and observed that oxidative dysregulation in p16-depleted cells was most profound in melanocytes, compared to keratinocytes or fibroblasts. Moreover, in the absence of p16 depletion or exogenous oxidative insult, melanocytes exhibited significantly higher basal levels of reactive oxygen species (ROS than these other epidermal cell types. Given the role of oxidative stress in melanoma development, we speculated that this increased susceptibility of melanocytes to oxidative stress (and greater reliance on p16 for suppression of ROS may explain why genetic compromise of p16 is more commonly associated with predisposition to melanoma rather than other cancers. Here we show that the presence of melanin accounts for this differential oxidative stress in normal and p16-depleted melanocytes. Thus the presence of melanin in the skin appears to be a double-edged sword: it protects melanocytes as well as neighboring keratinocytes in the skin through its capacity to absorb UV radiation, but its synthesis in melanocytes results in higher levels of intracellular ROS that may increase melanoma susceptibility.

  2. Heterogeneity of expression of epithelial-mesenchymal transition (EMT markers in melanocytes and melanoma cell lines

    Directory of Open Access Journals (Sweden)

    Ji Eun eKim

    2013-05-01

    Full Text Available The epithelial-mesenchymal transition (EMT describes a reversible switch from an epithelial-like to a mesenchymal-like phenotype. It is essential for the development of the normal epithelium and also contributes to the invasive properties of carcinomas. At the molecular level, the EMT transition is characterised by a series of coordinated changes including downregulation of the junctional protein E-cadherin (CDH1, up-regulation of transcriptional repressors of E-cadherin such as Snail (SNAI1 and Slug (SNAI2, and up-regulation of N-cadherin. We wished to determine whether cultured normal melanocytes and melanoma cell lines, which are derived from the neural crest, showed signs of a similarly coordinated phenotypic switch. We investigated normal melanocytes and 25 cell lines derived from New Zealand patients with metastatic melanoma. Most lines had been previously genotyped for common mutations such as BRAF, NRAS, PIK3CA, TP53 and CDKN2A. Expression of E-cadherin, N-cadherin, MITF, Snail, Slug, Axl, p53 and Hdm2 was compared by western blotting. Normal melanocytes expressed each of these proteins except for Snail, while normal melanocytes and almost every melanoma line expressed Slug. Expression of individual markers among different melanoma lines varied from high to low or undetectable. Quantitation of western blots showed that expression of MITF-M, the melanocyte-specific isoform of MITF, was positively related to that of E-cadherin but inversely related to that of N-cadherin and Axl. There was also no apparent relationship between expression of any particular marker and the presence of BRAF, NRAS, PIK3CA, TP53 or CDKN2A mutations. The results suggest that melanomas do not show the classical epithelial and mesenchymal phenotypes but rather display either high E-cadherin/high MITF-M expression on one hand, or high N-cadherin/high Axl expression on the other. These may correspond to differentiated and invasive phenotypes in vivo.

  3. [Phenotypic plasticity of neural crest-derived melanocytes and Schwann cells].

    Science.gov (United States)

    Dupin, Elisabeth

    2011-01-01

    Melanocytes, the pigmented cells of the skin, and the glial Schwann cells lining peripheral nerves are developmentally derived from an early and transient ectodermal structure of the vertebrate embryo, the neural crest, which is also at the origin of multiple neural and non-neural cell types. Besides melanocytes and neural cells of the peripheral nervous system, the neural crest cells give rise to mesenchymal cell types in the head, which form most of the craniofacial skeleton, dermis, fat tissue and vascular musculo-connective components. How such a wide diversity of differentiation fates is established during embryogenesis and is later maintained in adult tissues are among key questions in developmental and stem cell biology. The analysis of the developmental potentials of single neural crest cells cultured in vitro led to characterizing multipotent stem/progenitor cells as well as more restricted precursors in the early neural crest of avian and mammalian embryos. Data support a hierarchical model of the diversification of neural crest lineages through progressive restrictions of multipotent stem cell potentials driven by local environmental factors. In particular, melanocytes and glial Schwann cells were shown to arise from a common bipotent progenitor, which depends upon the peptide endothelin-3 for proliferation and self-renewal ability. In vivo, signaling by endothelin-3 and its receptor is also required for the early development of melanocytes and proper pigmentation of the vertebrate body. It is generally assumed that, after lineage specification and terminal differentiation, specialized cell types, like the melanocytes and Schwann cells, do not change their identity. However, this classic notion that somatic cell differentiation is a stable and irreversible process has been challenged by emerging evidence that dedifferentiation can occur in different biological systems through nuclear transfer, cell fusion, epigenetic modifications and ectopic gene

  4. Features of immunohistochemical diagnostic of melanocytic tumours

    Directory of Open Access Journals (Sweden)

    Shpon’ka I.S.

    2013-12-01

    Full Text Available Background. The malignant melanomas are the most important group of skin cancers. Although less common than the familiar basal and squamous cell tumours of the skin, they are much more frequently fatal, due to their intrinsic tendency to lymphatic and haematogenic metastasis. Objective. The article is devoted to parsing cases melanocytic tumours that were established through immunohistochemical study. Methods. In the study analyzed 236 patient material (150 women and 86 men aged 28 to 77 years during 2010-2013 turned out to clarify the histological diagnosis of skin tumors or metastases to lymph nodes (rare at other sites. The primary monoclonal antibodies used Сytokeratin, Рan Ab1 (clone AE1/AE3, S100 (clone 4C4.9, Ki-67 (clone SP6, Vimentin (clone V9, Melanoma gp100 (clone HMB-45. Results. Naevus proliferation rate showed a statistically significant difference with respect to proliferation rate of malignant melanomas (p<0,05. All samples (100% showed positive expression of high-intensity staining (+++ or moderate (++ intensity on the marker S100; 98,30% of samples (232 of 236 showed positive expression of marker HMB-45 at least in terms of tumor cells with intensity color from the high (+++ to weak (+ and 83.89% of the samples (198 of 236 were negative (– Сytokeratin, Рan Ab1 (other 38 cases showed weakly positive expression (+/– of tumor cells. Conclusions. 1. In the differential diagnosis of melanoma and naevus, we must bear in mind the uniformity immunophenotype of these tumors and consider only the cytological features of the tumor, changes in the structure of the epidermis and dermis (contour, symmetry, depth, inflammatory infiltration and proliferation rate. 2. Patients whose lymph nodes were the first clinical signs of cancer are always in need for additional immunohistochemical studies to avoid diagnostic errors. 3. The most common phenotype of melanocytic tumours responsible Сytokeratin, Рan–, Vimintin+, S100+, HMB-45

  5. Effects of Lutein and Zeaxanthin on LPS-Induced Secretion of IL-8 by Uveal Melanocytes and Relevant Signal Pathways

    OpenAIRE

    Shih-Chun Chao; Tommaso Vagaggini; Chan-Wei Nien; Sheng-Chieh Huang; Hung-Yu Lin

    2015-01-01

    The effects of lutein and zeaxanthin on lipopolysaccharide- (LPS-) induced secretion of IL-8 by uveal melanocytes (UM) were tested in cultured human UM. MTT assay revealed that LPS (0.01–1 μg/mL) and lutein and zeaxanthin (1–10 μM) did not influence the cell viability of cultured UM. LPS caused a dose-dependent increase of secretion of IL-8 by cultured UM. Lutein and zeaxanthin did not affect the constitutive secretion of IL-8. However, lutein and zeaxanthin decreased LPS-induced secretion of...

  6. Prediction of Dermoscopy Patterns for Recognition of both Melanocytic and Non-Melanocytic Skin Lesions

    Directory of Open Access Journals (Sweden)

    Qaisar Abbas

    2016-06-01

    Full Text Available A differentiation between all types of melanocytic and non-melanocytic skin lesions (MnM–SK is a challenging task for both computer-aided diagnosis (CAD and dermatologists due to the complex structure of patterns. The dermatologists are widely using pattern analysis as a first step with clinical attributes to recognize all categories of pigmented skin lesions (PSLs. To increase the diagnostic accuracy of CAD systems, a new pattern classification algorithm is proposed to predict skin lesions patterns by integrating the majority voting (MV–SVM scheme with multi-class support vector machine (SVM. The optimal color and texture features are also extracted from each region-of-interest (ROI dermoscopy image and then these normalized features are fed into an MV–SVM classifier to recognize seven classes. The overall system is evaluated using a dataset of 350 dermoscopy images (50 ROIs per class. On average, the sensitivity of 94%, specificity of 84%, 93% of accuracy and area under the receiver operating curve (AUC of 0.94 are achieved by the proposed MnM–SK system compared to state-of-the-art methods. The obtained result indicates that the MnM–SK system is successful for obtaining the high level of diagnostic accuracy. Thus, it can be used as an alternative pattern classification system to differentiate among all types of pigmented skin lesions (PSLs.

  7. Molecular cytogenetics of cutaneous melanocytic lesions - diagnostic, prognostic and therapeutic aspects

    NARCIS (Netherlands)

    Blokx, Willeke Am M.; van Dijk, Marcory C. R. F.; Ruiter, Dirk J.

    2010-01-01

    This review intends to update current knowledge regarding molecular cytogenetics in melanocytic tumours with a focus on cutaneous melanocytic lesions. Advantages and limitations of diverse, already established methods, such as (fluorescence) in situ hybridization and mutation analysis, to detect the

  8. Molecular cytogenetics of cutaneous melanocytic lesions - diagnostic, prognostic and therapeutic aspects.

    NARCIS (Netherlands)

    Blokx, W.A.M.; Dijk, M.C.R.F. van; Ruiter, D.J.

    2010-01-01

    This review intends to update current knowledge regarding molecular cytogenetics in melanocytic tumours with a focus on cutaneous melanocytic lesions. Advantages and limitations of diverse, already established methods, such as (fluorescence) in situ hybridization and mutation analysis, to detect the

  9. Does Melanoma Begin in a Melanocyte Stem Cell?

    Directory of Open Access Journals (Sweden)

    James D. Hoerter

    2012-01-01

    Full Text Available What is the cellular origin of melanoma? What role do melanocyte stem cells (MSC and other melanocyte precursors play in the development of melanoma? Are MSCs and other latent melanocyte precursors more susceptible to solar radiation? These and many other questions can be very effectively addressed using the zebrafish model. Zebrafish have a robust regenerative capability, permitting the study of how MSCs are regulated and recruited at specific times and places to generate the pigment pattern following fin amputation or melanocyte ablation. They can be used to determine the effects of environmental radiation on the proliferation, survival, repair, and differentiation of MSCs. Our lab is using zebrafish to investigate how UVA- (320–400 nm and UVB- (290–320 nm induced damage to MSCs may contribute to the development of melanoma. A review is given of MSCs in zebrafish as well as experimental techniques and drugs for manipulating MSC populations. These techniques can be used to design experiments to help answer many questions regarding the role of MSCs or melanocyte precursors in the formation of melanoma stem cells and tumors following exposure to UVA/UVB radiation.

  10. The Pure Culture of Fruiting Bodies of Scorias spongiosa,an Edible Mushroom of Bamboo%竹类食用蕈菌——海绵胶煤炱菌子实体的纯培养

    Institute of Scientific and Technical Information of China (English)

    贺新生; 刘超洋; 郑俊娟; 商圆圆; 霍存录

    2012-01-01

    The study explored the pure culture condition and development law of the fruiting bodies of Scorias spongiosa, in order to provide technical solutions for large - scale production of this new type of mushroom. More than 100 specimen of wild fruitbodies were collected from Zhejiang, Sichuan and Jiangsu province. Pure strains were obtained through tissus separation and pure culturing on agar, liquid and solid medium. Mycelium directly secreted gelatinous material to form colloidal fruitbodies in the agar and solid culture medium. In liquid medium, only mycelium and the mycelium sphere were formed, but not fruitbodies. The development process of this fungus was: conidia spore germination→ hyphal→colloid tissue→ branches of fruitbodies → small fruitbodies → formation pycnidium and spores liquid group → releasing spores. Under pure culture conditions it was difficult to form large fruiting bodies.%探索海绵胶煤炱菌子实体形成的培养条件,研究该菌的子实体发育规律,为规模化生产这种新型食用蕈菌提供技术方案.在浙江、四川、江苏等地共采集到100多份野生子实体标本,经组织分离得到纯培养菌株,在多种琼脂、液体、固体培养基上进行纯培养.在琼脂和固体培养基上,菌丝体直接分泌胶质物质形成胶质的小型子实体;在液体培养基上只形成菌丝和胶质菌丝球,不形成子实体.该菌的形态发育过程:分生孢子萌发→菌丝→胶质组织→子实体分枝→小型子实体→形成分生孢子器和分生孢子→释放孢子.海绵胶煤炱菌在纯培养条件下难以形成大型子实体.

  11. The epidermal melanocyte system in individuals of Scandinavian origin, determined by DOPA-staining and TEM

    DEFF Research Database (Denmark)

    Drzewiecki, K T; Piltz-Drzewiecka, J

    1979-01-01

    The quantitative evaluation of DOPA-positive epidermal melanocytes in 16 patients of Scandinavian origin showed both individual and regional differences in the melanocyte count. Our data is in agreement with other published studies. The distribution in the number of melanocytes varies significant...... concerning the presence of the epidermal melanin unit....

  12. File list: Oth.Epd.10.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Epd.10.AllAg.Melanocytes hg19 TFs and others Epidermis Melanocytes SRX382354,SR...X382353,SRX346921,SRX346922 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Epd.10.AllAg.Melanocytes.bed ...

  13. File list: InP.Epd.10.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Epd.10.AllAg.Melanocytes hg19 Input control Epidermis Melanocytes SRX382355,SRX...382356 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Epd.10.AllAg.Melanocytes.bed ...

  14. File list: DNS.Epd.10.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Epd.10.AllAg.Melanocytes hg19 DNase-seq Epidermis Melanocytes SRX089283,SRX0892...82,SRX201198,SRX201794,SRX201799,SRX100889 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Epd.10.AllAg.Melanocytes.bed ...

  15. File list: Oth.Epd.05.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Epd.05.AllAg.Melanocytes hg19 TFs and others Epidermis Melanocytes SRX346921,SR...X382354,SRX382353,SRX346922 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Epd.05.AllAg.Melanocytes.bed ...

  16. File list: His.Epd.10.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Epd.10.AllAg.Melanocytes hg19 Histone Epidermis Melanocytes SRX109440,SRX109442...,SRX109445,SRX109443,SRX109441,SRX109444 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Epd.10.AllAg.Melanocytes.bed ...

  17. File list: ALL.Epd.10.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Epd.10.AllAg.Melanocytes hg19 All antigens Epidermis Melanocytes SRX382354,SRX1...X109443,SRX100889,SRX382355,SRX382356,SRX109441,SRX109444 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Epd.10.AllAg.Melanocytes.bed ...

  18. File list: DNS.Epd.05.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Epd.05.AllAg.Melanocytes hg19 DNase-seq Epidermis Melanocytes SRX089283,SRX0892...82,SRX201198,SRX201794,SRX201799,SRX100889 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Epd.05.AllAg.Melanocytes.bed ...

  19. File list: InP.Epd.20.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Epd.20.AllAg.Melanocytes hg19 Input control Epidermis Melanocytes SRX382355,SRX...382356 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Epd.20.AllAg.Melanocytes.bed ...

  20. File list: ALL.Epd.05.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Epd.05.AllAg.Melanocytes hg19 All antigens Epidermis Melanocytes SRX346921,SRX3...X382356,SRX109441,SRX109445,SRX109443,SRX100889,SRX109444 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Epd.05.AllAg.Melanocytes.bed ...

  1. File list: DNS.Epd.50.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Epd.50.AllAg.Melanocytes hg19 DNase-seq Epidermis Melanocytes SRX089283,SRX2011...98,SRX089282,SRX201799,SRX201794 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Epd.50.AllAg.Melanocytes.bed ...

  2. File list: InP.Epd.50.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Epd.50.AllAg.Melanocytes hg19 Input control Epidermis Melanocytes SRX382355,SRX...382356 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Epd.50.AllAg.Melanocytes.bed ...

  3. File list: DNS.Epd.20.AllAg.Melanocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Epd.20.AllAg.Melanocytes hg19 DNase-seq Epidermis Melanocytes SRX089283,SRX2011...98,SRX089282,SRX201794,SRX201799,SRX100889 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Epd.20.AllAg.Melanocytes.bed ...

  4. Metaplastic breast carcinoma; melanocytic variant, a very rare tumour

    Science.gov (United States)

    Nzegwu, Martin A.; Sule, Emmanuel; Uzoigwe, Joseph; Achi, Franklyn

    2015-01-01

    Metaplastic breast carcinoma (MBC) is a rare heterogeneous malignancy, accounting for <1% of all invasive breast carcinomas, in which adenocarcinoma is found to coexist with an admixture of spindle, squamous, chondroid or bone-forming neoplastic cells. Melanocytic variant was first described by Ruffolo et al. in 1997. We report a case of MBC, melanocytic variant, in a 57-year-old Nigerian female who presented with a left breast mass 8 cm in diameter in the upper outer quadrant, hard and gradually increase in size to become painful. Breast examination showed gross asymmetry. Left breast was oedematous and shiny with extensive peau d'orange. No palpable axillary nodes were seen. Chest X-ray and abdominal ultrasound scan showed no involvement. Breast biopsy revealed an invasive metaplastic ductal carcinoma with melanocytic differentiation. PMID:25666364

  5. Differential sensitivity of aerobic gram-positive and gram-negative microorganisms to 2,4,6-trinitrotoluene (TNT) leads to dissimilar growth and TNT transformation: Results of soil and pure culture studies

    Energy Technology Data Exchange (ETDEWEB)

    Fuller, M.E.; Manning, J.F. Jr.

    1996-07-30

    The effects of 2,4,6-trinitrotoluene (TNT) on indigenous soil populations and pure bacterial cultures were examined. The number of colony-forming units (CFU) appearing when TNT-contaminated soil was spread on 0.3% molasses plates decreased by 50% when the agar was amended with 67 {mu}g TNT mL{sup -1}, whereas a 99% reduction was observed when uncontaminated soil was plated. Furthermore, TNT-contaminated soil harbored a greater number of organisms able to grow on plates amended with greater than 10 {mu}g TNT mL{sup -1}. The percentage of gram-positive isolates was markedly less in TNT-contaminated soil (7%; 2 of 30) than in uncontaminated soil (61%; 20 of 33). Pseudomonas aeruginosa, Pseudomonas corrugate, Pseudomonasfluorescens and Alcaligenes xylosoxidans made up the majority of the gram-negative isolates from TNT-contaminated soil. Gram-positive isolates from both soils demonstrated marked growth inhibition when greater than 8-16 {mu}g TNT mL{sup -1} was present in the culture media. Most pure cultures of known aerobic gram-negative organisms readily degraded TNT and evidenced net consumption of reduced metabolites. However, pure cultures of aerobic gram-positive bacteria were sensitive to relatively low concentrations of TNT as indicated by the 50% reduction in growth and TNT transformation which was observed at approximately 10 {mu}g TNT mL{sup -1}. Most non-sporeforming gram-positive organisms incubated in molasses media amended with 80 {mu}g TNT mL{sup -1} or greater became unculturable, whereas all strains tested remained culturable when incubated in mineral media amended with 98 {mu}g TNT mL{sup -1}, indicating that TNT sensitivity is likely linked to cell growth. These results indicate that gram-negative organisms are most likely responsible for any TNT transformation in contaminated soil, due to their relative insensitivity to high TNT concentrations and their ability to transform TNT.

  6. Astaxanthin and withaferin A block paracrine cytokine interactions between UVB-exposed human keratinocytes and human melanocytes via the attenuation of endothelin-1 secretion and its downstream intracellular signaling.

    Science.gov (United States)

    Niwano, Takao; Terazawa, Shuko; Nakajima, Hiroaki; Wakabayashi, Yuki; Imokawa, Genji

    2015-06-01

    Paracrine interactions between keratinocytes and melanocytes via cytokines play an essential role in regulating pigmentation in epidermal hyperpigmentary disorders. There is an urgent need for a human epidermal model in which melanogenic paracrine interactions between UVB-exposed keratinocytes and melanocytes can be precisely evaluated because human epidermal equivalents consisting of multilayered keratinocytes and melanocytes have significant limitations in this respect. To resolve this challenge, we established a co-culture system with cell inserts using human keratinocytes and human melanocytes that serves as an appropriate new model for UVB-induced hyperpigmentation. Using that new model, we examined the blocking effects of two natural chemicals, astaxanthin and withaferin A, on paracrine cytokine interactions between UVB-exposed keratinocytes and melanocytes and characterized their mechanisms of action. RT-PCR analysis showed that co-culture of human keratinocytes that had been exposed to UVB significantly stimulated human melanocytes to increase their expression of genes encoding microphthalmia-associated transcription factor, tyrosinase and tyrosinase-related protein 1. The catalytic activity of tyrosinase was also increased. ELISA assays revealed that UVB significantly increased the secretion of interleukin-1α, interleukin-6/8, granulocyte macrophage stimulatory factor and endothelin-1 but not α-melanocyte stimulating hormone. The addition of an endothelin-1 neutralizing antibody significantly abrogated the increase of tyrosinase activity. Post-irradiation treatment with astaxanthin or withaferin A significantly abolished the up-regulation of tyrosinase activity induced by UVB. Treatment with astaxanthin or withaferin A significantly reduced the increased levels of interleukin-1α, interleukin-6/8, granulocyte macrophage stimulatory factor and endothelin-1. Withaferin A but not astaxanthin also significantly abrogated the endothelin-1-stimulated activity

  7. Mutation in and lack of expression of tyrosinase-related protein-1 (TRP-1) in melanocytes from an individual with brown oculocutaneous albinism: A new subtype of albinism classified as {open_quotes}OCA3{close_quotes}

    Energy Technology Data Exchange (ETDEWEB)

    Boissy, R.E.; Zhao, H.; Austin, L.M.; Boissy, Y.L.; Zhao, Y. [Univ. of Cincinnati College of Medicine, OH (United States)] [and others

    1996-06-01

    Most types of human oculocutaneous albinism (OCA) result from mutations in the gene for tyrosinase (OCA1) or the P protein (OCA2), although other types of OCA have been described but have not been mapped to specific loci. Melanocytes were cultured from an African-American with OCA, who exhibited the phenotype of Brown OCA, and his normal fraternal twin. Melanocytes cultured from the patient with OCA and the normal twin appeared brown versus black, respectively. Melanocytes from both the patient with OCA and the normal twin demonstrated equal amounts of NP-40-soluble melanin; however, melanocytes from the patient with OCA contained only 7% of the amount of insoluble melanin found from the normal twin. Tyrosinase-related protein-1 (TRP-1) was not detected in the OCA melanocytes by use of various anti-TRP-1 probes. Furthermore, transcripts for TRP-1 were absent in cultured OCA melanocytes. The affected twin was homozygous for a single-bp deletion in exon 6, removing an A in codon 368 and leading to a premature stop at codon 384. Tyrosine hydroxylase activity of the OCA melanocytes was comparable to controls when assayed in cell lysates but was only 30% of controls when assayed in intact cells. We conclude that this mutation of the human TRP-1 gene affects its interaction with tyrosinase, resulting in dysregulation of tyrosinase activity, promotes the synthesis of brown versus black melanin, and is responsible for a third genetic type of OCA in humans, which we classify as {open_quotes}OCA3.{close_quotes} 69 refs., 7 figs., 3 tabs.

  8. 白介素-8对体外培养黑素细胞的影响%Effects of Interleukin-8 on Human Melanocytes in Vitro

    Institute of Scientific and Technical Information of China (English)

    陈静宇; 林新瑜; 王芳; 林鸿刚

    2012-01-01

    Objective: To establish the culture system of human normal melanocytes in vitro, and to study the effects of interleukin-8 at different concentrations on the proliferation of human melanocytes and melanin synthesis. Methods: MTT method and NaOH lysis method were employed to measure the effects of interleukin-8 at different concentrations on the proliferation of human melanocytes and melanin respectively. Apoptosis rates of melanocytes were detected with flow cytometer. Results: Interleukin-8 promotes melanocyte proliferation and melanin synthesis. Conclusions: Interleukin-8 enhances melanocyte proliferation and melanin synthesis.%目的:建立正常人表皮黑素细胞体外培养体系,观察不同浓度白介素-8对体外培养正常人黑素细胞的细胞增殖及黑素合成的影响.方法:采用四甲基偶氮唑盐(MTr)法测定不同浓度白介素-8对黑素细胞增值的影响,NaOH裂解法测定黑素生成量,流式细胞仪检测黑素细胞的凋亡率.结果:白介素-8对黑素细胞活力有增强作用,能使细胞增殖能力增加,黑素合成增加,但对黑素细胞的凋亡率无显著性影响.结论:白介素-8对体外培养的黑素细胞增殖及黑素生成都有增强作用.

  9. Congenital cerebriform melanocytic naevus with cutis verticis gyrata

    Directory of Open Access Journals (Sweden)

    Pai Vishaka

    2002-11-01

    Full Text Available Congenital melanocytic naevus is hamortomatous or non-neoplastic proliferation of abnormal mixture of tissue′s or non-neoplastic proliferation of abnormal mixture of tissue′s normal components. Cutis verticis gyrata is hypertrophy with parallel or gyrate folds of skin of scalp. We report a young man who presented to us with hair loss on the back of the head and increased growth of hair over the upper back since birth. A biopsy proved the diagnosis of congenital melanocytic naevus.

  10. Production of NO and N(inf2)O by Pure Cultures of Nitrifying and Denitrifying Bacteria during Changes in Aeration

    NARCIS (Netherlands)

    Kester, R.A.; De Boer, W.; Laanbroek, H.J.

    1997-01-01

    Peak emissions of NO and N2O are often observed after wetting of soil, The reactions to sudden changes in the aeration of cultures of nitrifying and denitrifying bacteria with respect to NO and N2O emissions were compared to obtain more information about the microbiological aspects of peak emissions

  11. Fermentation of Dietetic Fiber from Green Bean and Prickly Pear Shell by Pure and Mixture Culture of Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum 450B.

    Science.gov (United States)

    Mora-Cura, Y N; Meléndez-Rentería, N P; Delgado-García, M; Contreras-Esquivel, J C; Morlett-Chávez, J A; Aguilar, C N; Rodríguez-Herrera, R

    2017-03-23

    The aim of this study was to evaluate the fermentation of dietary fiber from green bean (Phaseolus vulgaris) and prickly pear shell (Opuntia ficus-indica) by Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum 450B growing as mono-culture and co-culture, the fermentation products, and proteins expressed during this process. The analysis of the fermentation profile showed a major growth of bacteria in the culture media of each dietary fiber supplemented with glucose, and particularly B. bifidum 450B at 48 h showed the highest growth. In the case of the co-culture, the growth was lower indicating the possible negative interaction between L. acidophilus LA-5 and B. bifidum 450B and may be due to the less amount of carbohydrates and the high content of non-soluble fiber that affected the nutrients availability for the bacterial strains. The pH changes indicated the presence of short-chain fatty acids (SCFAs), being acetate (46-100%) the main SCFA. Changes in the proteome concerned proteins that are involved in carbohydrate and other carbohydrate pathways. The characterization of the bacteria according to the growth, metabolites, and proteins expressed allows understanding the response to the change of environmental conditions and could be useful to understand L. acidophilus LA-5 and B. bifidum 450B strains' adaptation to specific applications.

  12. 胎儿头皮毛囊的黑素细胞定位及精细结构观察%Location and fine structure of melanocytes in human fetal scalp hair follicles

    Institute of Scientific and Technical Information of China (English)

    张汝芝; 朱文元; 李婉; 曹小芳; 江从军

    2009-01-01

    目的 探讨胎儿毛囊黑素细胞的定位及精细结构.方法 6个月胎儿因宫内发育畸形而引产、死亡后,取其带毛头皮,一部分常规包埋切片,分别用NKI/beteb、HMB-45、酪氨酸酶、酪氨酸酶相关蛋白1(TRP1)单抗染色.另一部分无菌处理后,0.1g/L的胶原酶Ⅱ和胰酶消化获得毛囊细胞,培养并传代后,透射电镜和原子力显微镜观察.结果 胎儿头皮毛囊NKI/beteb阳性细胞位于外根鞘,而在毛球内许多细胞HMB-45、酪氨酸酶、TRP1单抗染色阳性.在毛囊细胞的体外培养中,除去成纤维细胞和角质形成细胞后,可见两种黑素细胞,一种数目极少,色素很多,传代后消失;另一种数目较多,开始无色素,但增殖很快.传第3代后,几乎所有细胞NKI/beteb染色阳性.扫描电镜和原子力显微镜下,多数细胞为双极梭形,偶尔有3个树突.细胞体呈圆形或卵圆形,极突上无明显的分支,其内有少数散在的黑素体.结论 胎儿头皮毛囊外根鞘的黑素细胞推测为成黑素细胞和(或)其子代细胞.在早期的体外培养中,细胞增殖很快,但形态及功能上不成熟.%Objective To investigate the location,fine structure of melanocytes in human fetal scalp hair follicles.Methods The scalp with hair follicles was obtained from a dead fetus of 6 months of age,and divided into two parts.One part was embedded in paraffin,tissue sections were prepared with a width of 7 μm and stained with NKI/beteb,monoclonal antibodies to HMB-45,tyrosinase and tyrosinase-related protein 1(TRP1),respectively.The other part with hair follicles was treated with collagenase type Ⅱ 0.1 g/L and trypsin,then,cell suspension was collected and cultured.After 14-day culture,follicle melanocyte cells (FMC)were separated from keratinocytes by differential trypsinization,and fibroblasts were removed with geneticin.Following three times of pure passage,FMC were seeded and fixed on mica for scanning electron microscopy

  13. Updata on transplantion of melanocytes in the treatment of vitiligo%黑素细胞移植治疗白癜风的研究进展

    Institute of Scientific and Technical Information of China (English)

    秦晓明; 柯国琳; 杨孔超

    2015-01-01

    Transplantion of melanocytes is one of the methods in the treatment of vitiligo at stable stage, including cultured melanocyte transplantation and epidermal cells suspension transplantation. Co-culturing of melanocytes with adipose-derived stem cells may be a potential substitute for co-culturing with keratino-cytes. The efficiency of autologous melanocytes transplantation in vitiligo patients may be predicted by perile-sional skin-homing CD8+T cell activities and the dermal mesenchymal stem cells might be used as auxiliary agent to improve the efficacy. The progress on transplantion of melanocytes in the treatment of vitiligo is re-viewed.%黑素细胞移植是治疗稳定期白癜风的方法之一,主要包括培养的黑素细胞移植和表皮细胞悬液移植。近年来动物实验证实脂肪干细胞可代替角质形成细胞与黑素细胞共培养移植治疗白癜风。白癜风患者自体黑素细胞的移植效率可通过皮损周围皮肤归巢CD8+T细胞活动性来进行预测,真皮间充质干细胞可作为黑素细胞移植治疗白癜风的辅助剂。本文综述黑素细胞移植治疗白癜风的国内外进展,旨在为临床提供参考。

  14. Toxic influence of silver and uranium salts on activated sludge of wastewater treatment plants and synthetic activated sludge associates modeled on its pure cultures.

    Science.gov (United States)

    Tyupa, Dmitry V; Kalenov, Sergei V; Skladnev, Dmitry A; Khokhlachev, Nikolay S; Baurina, Marina M; Kuznetsov, Alexander Ye

    2015-01-01

    Toxic impact of silver and uranium salts on activated sludge of wastewater treatment facilities has been studied. Some dominating cultures (an active nitrogen fixer Agrobacterium tumifaciens (A.t) and micromyces such as Fusarium nivale, Fusarium oxysporum, and Penicillium glabrum) have been isolated and identified as a result of selection of the activated sludge microorganisms being steadiest under stressful conditions. For these cultures, the lethal doses of silver amounted 1, 600, 50, and 300 µg/l and the lethal doses of uranium were 120, 1,500, 1,000, and 1,000 mg/l, respectively. A.tumifaciens is shown to be more sensitive to heavy metals than micromyces. Synthetic granular activated sludge was formed on the basis of three cultures of the isolated micromyces steadiest against stress. Its granules were much more resistant to silver than the whole native activated sludge was. The concentration of silver causing 50 % inhibition of synthetic granular activated sludge growth reached 160-170 μg/l as far as for the native activated sludge it came only to 100-110 μg/l.

  15. Sixth form pure mathematics

    CERN Document Server

    Plumpton, C

    1968-01-01

    Sixth Form Pure Mathematics, Volume 1, Second Edition, is the first of a series of volumes on Pure Mathematics and Theoretical Mechanics for Sixth Form students whose aim is entrance into British and Commonwealth Universities or Technical Colleges. A knowledge of Pure Mathematics up to G.C.E. O-level is assumed and the subject is developed by a concentric treatment in which each new topic is used to illustrate ideas already treated. The major topics of Algebra, Calculus, Coordinate Geometry, and Trigonometry are developed together. This volume covers most of the Pure Mathematics required for t

  16. Endothelin-1 protects human melanocytes from UV-induced DNA damage by activating JNK and p38 signalling pathways.

    Science.gov (United States)

    von Koschembahr, Anne M; Swope, Viki B; Starner, Renny J; Abdel-Malek, Zalfa A

    2015-04-01

    Endothelin-1 is a paracrine factor with mitogenic, melanogenic and survival effects on cultured human melanocytes. We report that endothelin-1 signalling reduced the generation and enhanced the repair of ultraviolet radiation (UV)-induced DNA photoproducts, and inhibited apoptosis of human melanocytes, without increasing cAMP levels, melanin content or proliferation. Treatment with endothelin-1 activated the MAP kinases JNK and p38, as evidenced by phosphorylation of their target, activating transcription factor-2 (ATF-2). Endothelin-1 also enhanced the phosphorylation of JNK, p38 and ATF-2 by UV. The effects of endothelin-1 were dependent on increasing intracellular calcium mobilization by endothelin B receptor signalling. Activation of both JNK and p38 was required for reducing DNA photoproducts, but only JNK partially contributed to the survival effect of endothelin-1. ATF-2 activation depended mainly on JNK, yet was not sufficient for the effect of endothelin-1 on UV-induced DNA damage, suggesting the requirement for other JNK and p38 targets for this effect. Our results underscore the significance of endothelin-1 and endothelin B receptor signalling in reducing the genotoxic effects of UV via activating JNK and p38, hence restoring genomic stability of melanocytes.

  17. Rab11b mediates melanin transfer between donor melanocytes and acceptor keratinocytes via coupled exo/endocytosis.

    Science.gov (United States)

    Tarafder, Abul K; Bolasco, Giulia; Correia, Maria S; Pereira, Francisco J C; Iannone, Lucio; Hume, Alistair N; Kirkpatrick, Niall; Picardo, Mauro; Torrisi, Maria R; Rodrigues, Inês P; Ramalho, José S; Futter, Clare E; Barral, Duarte C; Seabra, Miguel C

    2014-04-01

    The transfer of melanin from melanocytes to keratinocytes is a crucial process underlying maintenance of skin pigmentation and photoprotection against UV damage. Here, we present evidence supporting coupled exocytosis of the melanin core, or melanocore, by melanocytes and subsequent endocytosis by keratinocytes as a predominant mechanism of melanin transfer. Electron microscopy analysis of human skin samples revealed three lines of evidence supporting this: (1) the presence of melanocores in the extracellular space; (2) within keratinocytes, melanin was surrounded by a single membrane; and (3) this membrane lacked the melanosomal membrane protein tyrosinase-related protein 1 (TYRP1). Moreover, co-culture of melanocytes and keratinocytes suggests that melanin exocytosis is specifically induced by keratinocytes. Furthermore, depletion of Rab11b, but not Rab27a, caused a marked decrease in both keratinocyte-stimulated melanin exocytosis and transfer to keratinocytes. Thus, we propose that the predominant mechanism of melanin transfer is keratinocyte-induced exocytosis, mediated by Rab11b through remodeling of the melanosome membrane, followed by subsequent endocytosis by keratinocytes.

  18. alpha-Melanocyte-stimulating-hormone precursors in the pig pituitary

    DEFF Research Database (Denmark)

    Fenger, M

    1986-01-01

    The occurrence of intermediates from the processing of ACTH-(1-39) [adrenocorticotropic hormone-(1-39)] to alpha-melanocyte-stimulating hormone was investigated in normal pig pituitaries by the use of sensitive and specific radioimmunoassays for ACTH-(1-13), ACTH-(1-14), ACTH-(1-13)-NH2 and ACTH-(1...

  19. Molecular biology of normal melanocytes and melanoma cells.

    Science.gov (United States)

    Bandarchi, Bizhan; Jabbari, Cyrus Aleksandre; Vedadi, Ali; Navab, Roya

    2013-08-01

    Malignant melanoma is one of the most aggressive malignancies in humans and is responsible for 60-80% of deaths from skin cancers. The 5-year survival of patients with metastatic malignant melanoma is about 14%. Its incidence has been increasing in the white population over the past two decades. The mechanisms leading to malignant transformation of melanocytes and melanocytic lesions are poorly understood. In developing malignant melanoma, there is a complex interaction of environmental and endogenous (genetic) factors, including: dysregulation of cell proliferation, programmed cell death (apoptosis) and cell-to-cell interactions. The understanding of genetic alterations in signalling pathways of primary and metastatic malignant melanoma and their interactions may lead to therapeutics modalities, including targeted therapies, particularly in advanced melanomas that have high mortality rates and are often resistant to chemotherapy and radiotherapy. Our knowledge regarding the molecular biology of malignant melanoma has been expanding. Even though several genes involved in melanocyte development may also be associated with melanoma cell development, it is still unclear how a normal melanocyte becomes a melanoma cell. This article reviews the molecular events and recent findings associated with malignant melanoma.

  20. Automatic Differential Diagnosis of Melanocytic Skin Tumors Using Ultrasound Data.

    Science.gov (United States)

    Andrėkutė, Kristina; Linkevičiūtė, Gintarė; Raišutis, Renaldas; Valiukevičienė, Skaidra; Makštienė, Jurgita

    2016-12-01

    We describe a novel automatic diagnostic system based on quantitative analysis of ultrasound data for differential diagnosis of melanocytic skin tumors. The proposed method has been tested on 160 ultrasound data sets (80 of malignant melanoma and 80 of benign melanocytic nevi). Acoustical, textural and shape features have been evaluated for each segmented lesion. Using parameters selected according to Mahalanobis distance and linear support vector machine classifier, we are able to differentiate malignant melanoma from benign melanocytic skin tumors with 82.4% accuracy (sensitivity = 85.8%, specificity = 79.6%). The results indicate that high-frequency ultrasound has the potential to be used for differential diagnosis of melanocytic skin tumors and to provide supplementary information on lesion penetration depth. The proposed system can be used as an additional tool for clinical decision support to improve the early-stage detection of malignant melanoma. Copyright © 2016 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.

  1. Optimal management of common acquired melanocytic nevi (moles: current perspectives

    Directory of Open Access Journals (Sweden)

    Sardana K

    2014-03-01

    Full Text Available Kabir Sardana, Payal Chakravarty, Khushbu Goel Department of Dermatology and STD, Maulana Azad Medical College and Lok Nayak Hospital, New Delhi, Delhi, India Abstract: Although common acquired melanocytic nevi are largely benign, they are probably one of the most common indications for cosmetic surgery encountered by dermatologists. With recent advances, noninvasive tools can largely determine the potential for malignancy, although they cannot supplant histology. Although surgical shave excision with its myriad modifications has been in vogue for decades, the lack of an adequate histological sample, the largely blind nature of the procedure, and the possibility of recurrence are persisting issues. Pigment-specific lasers were initially used in the Q-switched mode, which was based on the thermal relaxation time of the melanocyte (size 7 µm; 1 µsec, which is not the primary target in melanocytic nevus. The cluster of nevus cells (100 µm probably lends itself to treatment with a millisecond laser rather than a nanosecond laser. Thus, normal mode pigment-specific lasers and pulsed ablative lasers (CO2/erbium [Er]:yttrium aluminum garnet [YAG] are more suited to treat acquired melanocytic nevi. The complexities of treating this disorder can be overcome by following a structured approach by using lasers that achieve the appropriate depth to treat the three subtypes of nevi: junctional, compound, and dermal. Thus, junctional nevi respond to Q-switched/normal mode pigment lasers, where for the compound and dermal nevi, pulsed ablative laser (CO2/Er:YAG may be needed. If surgical excision is employed, a wide margin and proper depth must be ensured, which is skill dependent. A lifelong follow-up for recurrence and melanoma is warranted in predisposed individuals, although melanoma is decidedly uncommon in most acquired melanocytic nevi, even though histological markers may be seen on evaluation. Keywords: lasers, surgery, nevus, melanoma

  2. Rapid identification and quantification of Campylobacter coli and Campylobacter jejuni by real-time PCR in pure cultures and in complex samples

    Science.gov (United States)

    2011-01-01

    Background Campylobacter spp., especially Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli), are recognized as the leading human foodborne pathogens in developed countries. Livestock animals carrying Campylobacter pose an important risk for human contamination. Pigs are known to be frequently colonized with Campylobacter, especially C. coli, and to excrete high numbers of this pathogen in their faeces. Molecular tools, notably real-time PCR, provide an effective, rapid, and sensitive alternative to culture-based methods for the detection of C. coli and C. jejuni in various substrates. In order to serve as a diagnostic tool supporting Campylobacter epidemiology, we developed a quantitative real-time PCR method for species-specific detection and quantification of C. coli and C. jejuni directly in faecal, feed, and environmental samples. Results With a sensitivity of 10 genome copies and a linear range of seven to eight orders of magnitude, the C. coli and C. jejuni real-time PCR assays allowed a precise quantification of purified DNA from C. coli and C. jejuni. The assays were highly specific and showed a 6-log-linear dynamic range of quantification with a quantitative detection limit of approximately 2.5 × 102 CFU/g of faeces, 1.3 × 102 CFU/g of feed, and 1.0 × 103 CFU/m2 for the environmental samples. Compared to the results obtained by culture, both C. coli and C. jejuni real-time PCR assays exhibited a specificity of 96.2% with a kappa of 0.94 and 0.89 respectively. For faecal samples of experimentally infected pigs, the coefficients of correlation between the C. coli or C. jejuni real-time PCR assay and culture enumeration were R2 = 0.90 and R2 = 0.93 respectively. Conclusion The C. coli and C. jejuni real-time quantitative PCR assays developed in this study provide a method capable of directly detecting and quantifying C. coli and C. jejuni in faeces, feed, and environmental samples. These assays represent a new diagnostic tool for studying

  3. Comparison of the Transcriptional Profiles of Melanocytes from Dark and Light Skinned Individuals under Basal Conditions and Following Ultraviolet-B Irradiation.

    Science.gov (United States)

    López, Saioa; Smith-Zubiaga, Isabel; García de Galdeano, Alicia; Boyano, María Dolores; García, Oscar; Gardeazábal, Jesús; Martinez-Cadenas, Conrado; Izagirre, Neskuts; de la Rúa, Concepción; Alonso, Santos

    2015-01-01

    We analysed the whole-genome transcriptional profile of 6 cell lines of dark melanocytes (DM) and 6 of light melanocytes (LM) at basal conditions and after ultraviolet-B (UVB) radiation at different time points to investigate the mechanisms by which melanocytes protect human skin from the damaging effects of UVB. Further, we assessed the effect of different keratinocyte-conditioned media (KCM+ and KCM-) on melanocytes. Our results suggest that an interaction between ribosomal proteins and the P53 signaling pathway may occur in response to UVB in both DM and LM. We also observed that DM and LM show differentially expressed genes after irradiation, in particular at the first 6h after UVB. These are mainly associated with inflammatory reactions, cell survival or melanoma. Furthermore, the culture with KCM+ compared with KCM- had a noticeable effect on LM. This effect includes the activation of various signaling pathways such as the mTOR pathway, involved in the regulation of cell metabolism, growth, proliferation and survival. Finally, the comparison of the transcriptional profiles between LM and DM under basal conditions, and the application of natural selection tests in human populations allowed us to support the significant evolutionary role of MIF and ATP6V0B in the pigmentary phenotype.

  4. Interferon-gamma Inhibits Melanogenesis and Induces Apoptosis in Melanocytes: A Pivotal Role of CD8+ Cytotoxic T Lymphocytes in Vitiligo.

    Science.gov (United States)

    Yang, Lili; Wei, Yi; Sun, Yue; Shi, Weimin; Yang, Ji; Zhu, Lubing; Li, Ming

    2015-07-01

    Increased expression of the cytokine interferon (IFN)-γ plays a pivotal role in vitiligo-induced depigmentation. However, the major source of IFN-γ in vitiligo patients and the mechanisms underlying melanocyte destruction are unknown. In this study, a large number of skin infiltrating IFN-γ+ cells and CD8+ T cells were detected in progressive vitiligo. Among the peripheral blood mononuclear cells (PBMCs) of vitiligo patients, CD8+ cytotoxic T lymphocytes (CTLs) that express IFN-γ exhibited significant expansion, which suggests that activated CTLs are the main source of increased IFN-γ in progressive vitiligo. An in vitro analysis demonstrated that IFN-γ inhibits melanogenesis in primary cultured human melanocytes by altering melanogenic enzyme mRNA expression and, more importantly, that IFN-γ directly induces melanocyte apoptosis. Our data indicate that vitiligo pathophysiology may be linked to globally activated CD8+ CTL subpopulations, which produce increased IFN-γ and induce melanocyte dysfunction and apoptosis.

  5. Comparison of the Transcriptional Profiles of Melanocytes from Dark and Light Skinned Individuals under Basal Conditions and Following Ultraviolet-B Irradiation

    Science.gov (United States)

    López, Saioa; Smith-Zubiaga, Isabel; García de Galdeano, Alicia; Boyano, María Dolores; García, Oscar; Gardeazábal, Jesús; Martinez-Cadenas, Conrado; Izagirre, Neskuts; de la Rúa, Concepción; Alonso, Santos

    2015-01-01

    We analysed the whole-genome transcriptional profile of 6 cell lines of dark melanocytes (DM) and 6 of light melanocytes (LM) at basal conditions and after ultraviolet-B (UVB) radiation at different time points to investigate the mechanisms by which melanocytes protect human skin from the damaging effects of UVB. Further, we assessed the effect of different keratinocyte-conditioned media (KCM+ and KCM-) on melanocytes. Our results suggest that an interaction between ribosomal proteins and the P53 signaling pathway may occur in response to UVB in both DM and LM. We also observed that DM and LM show differentially expressed genes after irradiation, in particular at the first 6h after UVB. These are mainly associated with inflammatory reactions, cell survival or melanoma. Furthermore, the culture with KCM+ compared with KCM- had a noticeable effect on LM. This effect includes the activation of various signaling pathways such as the mTOR pathway, involved in the regulation of cell metabolism, growth, proliferation and survival. Finally, the comparison of the transcriptional profiles between LM and DM under basal conditions, and the application of natural selection tests in human populations allowed us to support the significant evolutionary role of MIF and ATP6V0B in the pigmentary phenotype. PMID:26244334

  6. Altered E-Cadherin Levels and Distribution in Melanocytes Precede Clinical Manifestations of Vitiligo.

    Science.gov (United States)

    Wagner, Roselyne Y; Luciani, Flavie; Cario-André, Muriel; Rubod, Alain; Petit, Valérie; Benzekri, Laila; Ezzedine, Khaled; Lepreux, Sébastien; Steingrimsson, Eirikur; Taieb, A; Gauthier, Yvon; Larue, Lionel; Delmas, Véronique

    2015-07-01

    Vitiligo is the most common depigmenting disorder resulting from the loss of melanocytes from the basal epidermal layer. The pathogenesis of the disease is likely multifactorial and involves autoimmune causes, as well as oxidative and mechanical stress. It is important to identify early events in vitiligo to clarify pathogenesis, improve diagnosis, and inform therapy. Here, we show that E-cadherin (Ecad), which mediates the adhesion between melanocytes and keratinocytes in the epidermis, is absent from or discontinuously distributed across melanocyte membranes of vitiligo patients long before clinical lesions appear. This abnormality is associated with the detachment of the melanocytes from the basal to the suprabasal layers in the epidermis. Using human epidermal reconstructed skin and mouse models with normal or defective Ecad expression in melanocytes, we demonstrated that Ecad is required for melanocyte adhesiveness to the basal layer under oxidative and mechanical stress, establishing a link between silent/preclinical, cell-autonomous defects in vitiligo melanocytes and known environmental stressors accelerating disease expression. Our results implicate a primary predisposing skin defect affecting melanocyte adhesiveness that, under stress conditions, leads to disappearance of melanocytes and clinical vitiligo. Melanocyte adhesiveness is thus a potential target for therapy aiming at disease stabilization.

  7. Rehabilitation of pure alexia

    DEFF Research Database (Denmark)

    Starrfelt, Randi; Ólafsdóttir, Rannveig Rós; Arendt, Ida-Marie

    2013-01-01

    Acquired reading problems caused by brain injury (alexia) are common, either as a part of an aphasic syndrome, or as an isolated symptom. In pure alexia, reading is impaired while other language functions, including writing, are spared. Being in many ways a simple syndrome, one would think...... that pure alexia was an easy target for rehabilitation efforts. We review the literature on rehabilitation of pure alexia from 1990 to the present, and find that patients differ widely on several dimensions like alexia severity, and associated deficits. Many patients reported to have pure alexia...... in the reviewed studies, have associated deficits like agraphia or aphasia and thus do not strictly conform to the diagnosis. Few studies report clear and generalisable effects of training, none report control data, and in many cases the reported findings are not supported by statistics. We can, however...

  8. Rehabilitation of pure alexia

    DEFF Research Database (Denmark)

    Starrfelt, Randi; Ólafsdóttir, Rannveig Rós; Arendt, Ida-Marie

    2013-01-01

    that pure alexia was an easy target for rehabilitation efforts. We review the literature on rehabilitation of pure alexia from 1990 to the present, and find that patients differ widely on several dimensions like alexia severity, and associated deficits. Many patients reported to have pure alexia......Acquired reading problems caused by brain injury (alexia) are common, either as a part of an aphasic syndrome, or as an isolated symptom. In pure alexia, reading is impaired while other language functions, including writing, are spared. Being in many ways a simple syndrome, one would think...... in the reviewed studies, have associated deficits like agraphia or aphasia and thus do not strictly conform to the diagnosis. Few studies report clear and generalisable effects of training, none report control data, and in many cases the reported findings are not supported by statistics. We can, however...

  9. Cellular origin and developmental mechanisms during the formation of skin melanocytes

    Energy Technology Data Exchange (ETDEWEB)

    Ernfors, Patrik, E-mail: patrik.ernfors@ki.se [Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, 17177 Stockholm (Sweden)

    2010-05-01

    Melanocytes are derived from the neural crest (NC), which are transient multipotent cells arising by delamination from the developing dorsal neural tube. During recent years, signaling systems and molecular mechanisms of melanocyte development have been studied in detail, but the exact diversification of the NC into melanocytes and how they migrate, expand and disperse in the skin have not been fully understood. The recent finding that Schwann cell precursors (SCPs) of the growing nerve represents a stem cell niche from which various cell types, including Schwann cells, endoneural fibroblasts and melanocytes arise has exposed new knowledge on the cellular basis for melanocyte development. This opens for the identification of new factors and reinterpretation of old data on cell fate instructive, proliferative, survival and cell homing factors participating in melanocyte development.

  10. Paracrine Secreted Frizzled-Related Protein 4 Inhibits Melanocytes Differentiation in Hair Follicle

    Science.gov (United States)

    Guo, Haiying; Lei, Mingxing; Li, Yuhong; Liu, Yingxin; Tang, Yinhong; Xing, Yizhan; Deng, Fang

    2017-01-01

    Wnt signaling plays crucial role in regulating melanocyte stem cells/melanocyte differentiation in the hair follicle. However, how the Wnt signaling is balanced to be overactivated to control follicular melanocytes behavior remains unknown. Here, by using immunofluorescence staining, we showed that secreted frizzled-related protein 4 (sFRP4) is preferentially expressed in the skin epidermal cells rather than in melanocytes. By overexpression of sFRP4 in skin cells in vivo and in vitro, we found that sFRP4 attenuates activation of Wnt signaling, resulting in decrease of melanocytes differentiation in the regenerating hair follicle. Our findings unveiled a new regulator that involves modulating melanocytes differentiation through a paracrine mechanism in hair follicle, supplying a hope for potential therapeutic application to treat skin pigmentation disorders. PMID:28337220

  11. 石油降解单菌株及混合菌降解产物分析%Analysis biodegradation products of oil compounds by pure and mixed bacterial cultures

    Institute of Scientific and Technical Information of China (English)

    花莉; 彭香玉; 范洋; 解井坤

    2014-01-01

    Four oil-degrading bacteria strains ,isolated from oil-contaminated soil of Chan-gqing Oilfied Company ,were mixed to construct bacterial consortiums for crude oil degrada-tion .Isolated microorganisms ,in the form of pure and mixed ,were cultured for 5 days at 30 ℃ in a medium containing diesel .These pure bacteria degrade 41 .83% ~54 .87% of diesel and among them Klebsiella variicola showed the greatest capability for degradation diesel . Biodegradation of diesel by the mixed culture of four bacteria was 64 .27% .The degradability of bacterial mixed culture is higher than the four pure cultures .Raoultella planticola and K lebsiella v ariicola can degrade branched-alkene ,Serratia m arcescens and Bacillus cereus can oxidative unsaturated aldehyde to acid .Bacillus cereus and K lebsiella v ariicola trans-formed cycloalkanes into linear ester via a primary alcohol .In addition K lebsiella v ariicola degraded a triple bond between two carbon atoms (C≡C) of 7-Pentadecyne .It was a promi-nent strain in the oil degradation process .Microbial antagonism in four strains made them not suitable for degrading cycloalkanes .The research provides theoretical foundation for the study on metabolism of hydrocarbons and screen for functional microorganism .%从中国石油长庆石化公司附近的油泥中分离筛选出4株石油降解菌,用于组建降解原油的混合菌体系.在等接种量培养条件下,单菌株对石油烃的降解率达到41.83%~54.87%,而混合菌降解效率高于单菌株,达到64.27%.不同微生物在降解过程中起着不同的作用.居植物柔武士菌(Raoultella p lanticola)具有脱烷基功能,对一些支链烃有降解效果;蜡状芽孢杆菌(Bacillus cereus)既可以氧化末端烯烃,又可以降解环烷烃;克雷伯氏菌(K lebsiella v ari-icola)是优势菌种,不仅具有以上3种降解功能,还可以降解炔烃;粘质沙雷氏菌(Serratia marcescens

  12. Nevus depigmentosus treated by melanocyte-keratinocyte transplantation

    Directory of Open Access Journals (Sweden)

    Sanjeev V Mulekar

    2011-01-01

    Full Text Available Background : Nevus depigmentosus is a congenital, nonprogressive hypopigmented disorder. Various therapeutic methods have been attempted to repigment nevus with variable results. Objective : The objective of this study is to report our experience of treatment of nevus depigmentosus with a combination of noncultured melanocyte-keratinocyte transplantation (MKTP and excimer laser sessions. Materials and Methods : Six patients (male 1, female 5 of nevus depigmentosus were treated with a combination of noncultured melanocyte-keratinocyte transplantation and excimer laser. One patient was lost to follow-up. Remaining five patients were observed for a period ranging from 7 to 30 months. Results : Two patients responded poorly to MKTP. The remaining three patients responded with repigmentation ranging from 80% to 100% but the quality of repigmentation was unsatisfactory in two of them. Conclusion : Though repigmentation of nevus depigmentosus is possible by grafting techniques, the results are inconsistent and recurrence is possible.

  13. The optical properties and spectral features of malignant skin melanocytes in the terahertz frequency range

    Science.gov (United States)

    Goryachuk, A. A.; Begaeva, V. A.; Khodzitsky, M. K.; Truloff, A. S.

    2016-08-01

    The samples of cells of mice's melanocytes have been investigated. Their optical properties and spectral features were investigated by terahertz time-domain spectroscopy (TDS) in transmission mode. It was found that the optical properties of oncological melanocytes and normal cells are different and oncological cells have spectral features of absorption coefficient so it can be concluded that it is easy to discriminate mice's oncological skin melanocytes by using THz TDS.

  14. Impression cytology of melanocytic conjunctival tumours using the Biopore membrane.

    Science.gov (United States)

    Keijser, S; Missotten, G S; De Wolff-Rouendaal, D; Verbeke, S L J; Van Luijk, C M; Veselic-Charvat, M; de Keizer, R J W

    2007-01-01

    To compare a new Biopore membrane impression cytology method with the routinely used exfoliative cytology in patients with a melanocytic lesion of the conjunctiva. Sixty-eight consecutive patients with a conjunctival melanocytic lesion underwent Biopore membrane impression cytology as well as exfoliative cytology. A histologic sample was also available in 26 cases. All Biopore samples were stained immediately with RAL 555. Both Biopore and exfoliative cytology samples were assessed by two cytopathologists and graded into four different categories of atypia. Twenty-three out of 26 Biopores and 20 out of 24 for the exfoliative smears correlated with the corresponding histologic sample. Biopore cytology resulted in higher numbers of cells with a greater density compared to exfoliative cytology. Biopore cytology can be used for cytologic sampling of conjunctival melanocytic lesions. Because of the larger amount and higher density of cells obtained with the Biopore membrane, interpretation by a pathologist is easier and faster. Sampling of the fornix, caruncula, and ocular material in children is difficult with the Biopore method, and exfoliative cytology seems to be the favorable test in those situations.

  15. Malignant neuroectodermal tumor with melanocytic and rhabdomyoblastic differentiation

    Directory of Open Access Journals (Sweden)

    Brian P. Rubin

    2009-12-01

    Full Text Available Malignant melanoma can metastasize widely and vary significantly in its histological appearance; it rarely presents as a deep-seated mass without an obvious primary site elsewhere. Malignant peripheral nerve sheath tumor (MPNST is a high-grade sarcoma characterized by conventional and epithelioid subtypes. MPNST can demonstrate heterologous differentiation, usually in the form of osteosarcomatous, chondrosarcomatous, or rhabdo-myosarcomatous differentiation. MPNST does not harbor true melanocytic differentiation, although epithelioid MPNST typically is diffusely S-100 protein positive and superficially can resemble malignant melanoma. An unusual intra-abdominal mass was recently encountered with features of both melanoma and conventional or epithelioid MPNST containing a fascicular spindle cell component, an epithelioid component with melanocytic differentiation, as well as a rhabdomyosarcomatous component. The terminology “malignant neuroectodermal tumor with melanocytic and rhabdomyoblastic differentiation” is proposed to describe this neoplasm, reflecting the unusual concomittant lines of differentiation as well as offering a possible rationale for nosologically challenging aspects of this neoplasm.

  16. Ultrastructural changes in the melanocytes of aging human choroid.

    Science.gov (United States)

    Nag, Tapas Chandra

    2015-12-01

    Retinal pigment epithelial cells as well as choroidal melanocytes (CM) possess melanin granules. The former show clear, age-related changes (formation of lipofuscin granules with a concomitant decrease in melanin content); however, data on changes in the CM with aging are fairly limited. We examined CM in human macular and mid-peripheral areas by light- and transmission electron microscopy in 50-94 year-old donor eyes (N=12). Unlike in the choroid of lower ages, the melanocytes from aging choroid (>75 years) showed partial fusion of about 8-15 melanosomes, forming rosettes-like structures. Besides, there was evidence of emptiness in cytoplasm caused by the loss of melanosomes in aged CM, as was confirmed by quantification in macular part of choroid. In advanced aged eyes (85-94-year-old), the CM possessed many lipid droplets as well as irregular lipofuscin granules, the latter had a tendency to fuse with melanosomes, as happens in aged retinal pigment epithelium. Macrophages in their cytoplasm contained abundant irregular as well as clumped melanosomes of variable size, suggesting that damaged granules/melanocytes are cleared by these phagocytes. These obvious changes in the CM are likely to make the choroid prone to damage by visible light.

  17. Melanoma and melanocytic nevi in decorative tattoos: three case reports.

    Science.gov (United States)

    Varga, Erika; Korom, Irma; Varga, János; Kohán, József; Kemény, Lajos; Oláh, Judit

    2011-12-01

    In response to the demands of style and fashion, the number of decorative tattoos has been increasing worldwide. This has been paralleled by a rising incidence of melanocytic proliferations, including melanoma. The coincidence of various dermatological diseases and skin tumors with tattoos has been documented with some frequency, but reports of melanoma associated with tattoos are exceedingly rare. To date, only 13 cases have been documented in the English language literature. The possibility of an association between melanocytic proliferations and tattoos remains an area for further study. This report presents two cases of melanocytic nevi and one of melanoma occurring in association with a decorative tattoos. At present, the pathogenesis of melanoma developing in a tattoo is unknown. Mere coincidence cannot be ruled out. However, trauma, ultraviolet light exposure, a photoallergic effect, or an inflammatory reaction may promote malignant transformation. Clinicians and histopathologists should be aware of the clinical and pathological features if they are to make a correct diagnosis. Copyright © 2011 John Wiley & Sons A/S.

  18. Gene expression profiling of melanocytes following Q-Switched Ruby laser irradiation.

    Science.gov (United States)

    Hafner, Christian; Stempfl, Thomas; Bäumler, Wolfgang; Hohenleutner, Ulrich; Landthaler, Michael; Vogt, Thomas

    2008-01-01

    The Q-switched Ruby laser (QSRL) is used for the treatment of pigmented lesions. The influence of QSRL treatment on gene expression of nontransformed primary melanocytes has not been addressed in vitro. We investigated the gene expression profile of melanocytes following QSRL irradiation. Primary melanocytes were irradiated with the QSRL (694 nm). Early and late transcriptional effects were analyzed using the Affymetrix gene array platform. Laser irradiation of melanocytes had minor effects on mRNA expression. We found only 31 out of 14,500 genes which were at least twofold up- or downregulated. The differential expression of heme oxygenase 1 and galanin in QSRL-treated melanocytes was additionally confirmed by real-time RT-PCR. Analysis of a selection of 36 genes which are known to be associated with malignant melanoma development and progression revealed no significantly aberrant expression in the QSRL-treated melanocytes. Our study shows that QSRL treatment of primary melanocytes in vitro does not cause major alterations of global gene expression and particularly of genes associated with malignant melanoma. However, since QSRL treatment may have different effects on gene expression of melanocytic cells in vivo, further studies are required to evaluate QSRL treatment of (nevo-) melanocytic lesions. (c) 2008 S. Karger AG, Basel.

  19. Dysregulated autophagy increased melanocyte sensitivity to H2O2-induced oxidative stress in vitiligo

    Science.gov (United States)

    He, Yuanmin; Li, Shuli; Zhang, Weigang; Dai, Wei; Cui, Tingting; Wang, Gang; Gao, Tianwen; Li, Chunying

    2017-01-01

    In vitiligo, melanocytes are particularly vulnerable to oxidative stress owing to the pro-oxidant state generated during melanin synthesis and to the genetic antioxidant defects. Autophagy is a controlled self-digestion process which can protect cells against oxidative damage. However, the exact role of autophagy in vitiligo melanocytes in response to oxidative stress and the mechanism involved are still not clear. To determine the implications of autophagy for melanocyte survival in response to oxidative stress, we first detected the autophagic flux in normal melanocytes exposure to H2O2, and found that autophagy was significantly enhanced in normal melanocytes, for protecting cells against H2O2-induced oxidative damage. Nevertheless, vitiligo melanocytes exhibited dysregulated autophagy and hypersensitivity to H2O2-induced oxidative injury. In addition, we confirmed that the impairment of Nrf2-p62 pathway is responsible for the defects of autophagy in vitiligo melanocytes. Noteworthily, upregulation of the Nrf2-p62 pathway or p62 reduced H2O2-induced oxidative damage of vitiligo melanocytes. Therefore, our data demonstrated that dysregulated autophagy owing to the impairment of Nrf2-p62 pathway increase the sensitivity of vitiligo melanocytes to oxidative stress, thus promote the development of vitiligo. Upregulation of p62-dependent autophagy may be applied to vitiligo treatment in the future. PMID:28186139

  20. Melanocytes in the skin--comparative whole transcriptome analysis of main skin cell types.

    Science.gov (United States)

    Reemann, Paula; Reimann, Ene; Ilmjärv, Sten; Porosaar, Orm; Silm, Helgi; Jaks, Viljar; Vasar, Eero; Kingo, Külli; Kõks, Sulev

    2014-01-01

    Melanocytes possess several functions besides a role in pigment synthesis, but detailed characteristics of the cells are still unclear. We used whole transcriptome sequencing (RNA-Seq) to assess differential gene expression of cultivated normal human melanocytes with respect to keratinocytes, fibroblasts and whole skin. The present results reveal cultivated melanocytes as highly proliferative cells with possible stem cell-like properties. The enhanced readiness to regenerate makes melanocytes the most vulnerable cells in the skin and explains their high risk of developing into malignant melanoma.

  1. Pure-tone Audiometer

    Science.gov (United States)

    Kapul, A. A.; Zubova, E. I.; Torgaev, S. N.; Drobchik, V. V.

    2017-08-01

    The research focuses on a pure-tone audiometer designing. The relevance of the study is proved by high incidence of an auditory analyser in older people and children. At first, the article provides information about subjective and objective audiometry methods. Secondly, we offer block-diagram and basic-circuit arrangement of device. We decided to base on STM32F407VG microcontroller and use digital pot in the function of attenuator. Third, we implemented microcontroller and PC connection. C programming language is used for microcontroller’s program and PC’s interface. Fourthly, we created the pure-tone audiometer prototype. In the future, we will implement the objective method ASSR in addition to pure-tone audiometry.

  2. Aqueous humor tyrosinase activity is indicative of iris melanocyte toxicity.

    Science.gov (United States)

    Mahanty, Sarmistha; Kawali, Ankush A; Dakappa, Shruthi Shirur; Mahendradas, Padmamalini; Kurian, Mathew; Kharbanda, Varun; Shetty, Rohit; Setty, Subba Rao Gangi

    2017-09-01

    Antibiotics such as fluoroquinolones (FQLs) are commonly used to treat ocular infections but are also known to cause dermal melanocyte toxicity. The release of dispersed pigments from the iris into the aqueous humor has been considered a possible ocular side effect of the systemic administration of FQLs such as Moxifloxacin, and this condition is known as bilateral acute iris transillumination (BAIT). Bilateral acute depigmentation of iris (BADI) is a similar condition, with iris pigment released into the aqueous, but it has not been reported as a side effect of FQL. Iris pigments are synthesized by the melanogenic enzyme tyrosinase (TYR) and can be detected but not quantified by using slit-lamp biomicroscopy. The correlation between dispersed pigments in the aqueous and the extent of melanocyte toxicity due to topical antibiotics in vivo is not well studied. Here, we aimed to study the effect of topical FQLs on iris tissue, the pigment release in the aqueous humor and the development of clinically evident iris atrophic changes. We evaluated this process by measuring the activity of TYR in the aqueous humor of 82 healthy eyes undergoing cataract surgery following topical application of FQLs such as Moxifloxacin (27 eyes, preservative-free) or Ciprofloxacin (29 eyes, with preservative) or the application of non-FQL Tobramycin (26 eyes, with preservative) as a control. In addition, the patients were questioned and examined for ocular side effects in pre- and post-operative periods. Our data showed a significantly higher mean TYR activity in the aqueous humor of Ciprofloxacin-treated eyes compared to Moxifloxacin- (preservative free, p humor from both Ciprofloxacin- and Moxifloxacin-treated eyes showed the presence of soluble TYR enzyme, thus reflecting its toxicity to iris melanocytes and corresponding to its activity in the aqueous humor. Intriguingly, none of these patients developed any clinically appreciable ocular side effects characteristic of BAIT or BADI

  3. Narrow Band Ultraviolet B Treatment for Human Vitiligo Is Associated with Proliferation, Migration, and Differentiation of Melanocyte Precursors.

    Science.gov (United States)

    Goldstein, Nathaniel B; Koster, Maranke I; Hoaglin, Laura G; Spoelstra, Nicole S; Kechris, Katerina J; Robinson, Steven E; Robinson, William A; Roop, Dennis R; Norris, David A; Birlea, Stanca A

    2015-08-01

    In vitiligo, the autoimmune destruction of epidermal melanocytes produces white spots that can be repigmented by melanocyte precursors from the hair follicles, following stimulation with UV light. We examined by immunofluorescence the distribution of melanocyte markers (C-KIT, DCT, PAX3, and TYR) coupled with markers of proliferation (KI-67) and migration (MCAM) in precursors and mature melanocytes from the hair follicle and the epidermis of untreated and narrow band UVB (NBUVB)-treated human vitiligo skin. NBUVB was associated with a significant increase in the number of melanocytes in the infundibulum and with restoration of the normal melanocyte population in the epidermis, which was lacking in the untreated vitiligo. We identified several precursor populations (melanocyte stem cells, melanoblasts, and other immature phenotypes), and progressively differentiating melanocytes, some with putative migratory and/or proliferative abilities. The primary melanocyte germ was present in the untreated and treated hair follicle bulge, whereas a possible secondary melanocyte germ composed of C-KIT+ melanocytes was found in the infundibulum and interfollicular epidermis of UV-treated vitiligo. This is an exceptional model for studying the mobilization of melanocyte stem cells in human skin. Improved understanding of this process is essential for designing better treatments for vitiligo, ultimately based on melanocyte stem cell activation and mobilization.

  4. Plexin B1 inhibits MET through direct association and regulates Shp2 expression in melanocytes.

    Science.gov (United States)

    Soong, Joanne; Scott, Glynis

    2013-01-15

    Plexin B1, the receptor for Semaphorin 4D (Sema4D), is expressed by melanocytes in the skin. We recently showed that Sema4D suppresses activation of the hepatocyte growth factor receptor, MET, in melanocytes, and that knockdown of Plexin B1 results in activation of MET. MET signaling mediates proliferation, survival and migration in melanocytes, and its activation is associated with transformation of melanocytes to melanoma. In this report we investigated the mechanism by which Plexin B1 inhibits MET activation. Our results show that Plexin B1 and MET exist as an oligomeric receptor-receptor complex in melanocytes, and that receptor association is increased by Sema4D. MET and Plexin B1 receptor complexes were identified along the cell body of melanocytes, and Sema4D increased receptor association on dendrites, suggesting that Sema4D regulates MET-dependent processes at precise locations on the melanocyte. Despite activation of MET, Plexin B1 knockdowns proliferated slowly and showed increased apoptosis compared with controls. Shp2, a non-receptor protein tyrosine phosphatase, translates growth and survival signals from MET and other receptor tyrosine kinases. Plexin B1 knockdowns had markedly lower levels of Shp2 compared with controls, and Sema4D upregulated Shp2 expression at the protein and message level in normal melanocytes. Functional studies showed that blockade of Shp2 activity abrogated MET-dependent activation of Erk1/Erk2 and Akt in melanocytes. These results suggest a complex role for Sema4D and Plexin B1 in orchestrating signaling from the MET receptor in melanocytes. Because Shp2 is a downstream adaptor protein for multiple receptors, Sema4D may control the effects of several growth factors on melanocytes through regulation of Shp2.

  5. Heterogeneous topographic profiles of kinetic and cell cycle regulator microsatellites in atypical (dysplastic) melanocytic nevi.

    Science.gov (United States)

    Husain, Ehab A; Mein, Charles; Pozo, Lucia; Blanes, Alfredo; Diaz-Cano, Salvador J

    2011-04-01

    Atypical (dysplastic) melanocytic nevi are clinically heterogeneous malignant melanoma precursors, for which no topographic analysis of cell kinetic, cell cycle regulators and microsatellite profile is available. We selected low-grade atypical melanocytic nevi (92), high-grade atypical melanocytic nevi (41), melanocytic nevi (18 junctional, 25 compound) and malignant melanomas (16 radial growth phase and 27 vertical growth phase). TP53, CDKN2A, CDKN1A, and CDKN1B microsatellite patterns were topographically studied after microdissection; Ki-67, TP53, CDKN2A, CDKN1A, and CDKN1B expressions and DNA fragmentation by in situ end labeling for apoptosis were topographically scored. Results were statistically analyzed. A decreasing junctional-dermal marker expression gradient was observed, directly correlating with atypical melanocytic nevus grading. High-grade atypical melanocytic nevi revealed coexistent TP53-CDKN2A-CDKN1B microsatellite abnormalities, and significantly higher junctional Ki67-TP53 expression (inversely correlated with CDKN1A-CDKN1B expression and in situ end labeling). Malignant melanomas showed coexistent microsatellite abnormalities (CDKN2A-CDKN1B), no topographic gradient, and significantly decreased expression. Melanocytic nevi and low-grade atypical melanocytic nevi revealed sporadic junctional CDKN2A microsatellite abnormalities and no significant topographic kinetic differences. High-grade atypical melanocytic nevi accumulate junctional TP53-CDKN1A-CDKN1B microsatellite abnormalities, being progression TP53-independent and better assessed in the dermis. Melanocytic nevi and low-grade atypical melanocytic nevi show low incidence of microsatellite abnormalities, and kinetic features that make progression unlikely.

  6. Transforming growth factor beta1 regulates melanocyte proliferation and differentiation in mouse neural crest cells via stem cell factor/KIT signaling.

    Science.gov (United States)

    Kawakami, Tamihiro; Soma, Yoshinao; Kawa, Yoko; Ito, Masaru; Yamasaki, Emiko; Watabe, Hidenori; Hosaka, Eri; Yajima, Kenji; Ohsumi, Kayoko; Mizoguchi, Masako

    2002-03-01

    Stem cell factor is essential to the migration and differentiation of melanocytes during embryogenesis based on the observation that mutations in either the stem cell factor gene, or its ligand, KIT, result in defects in coat pigmentation in mice. Stem cell factor is also required for the survival of melanocyte precursors while they are migrating towards the skin. Transforming growth factor beta1 has been implicated in the regulation of both cellular proliferation and differentiation. NCC-melb4, an immortal cloned cell line, was cloned from a mouse neural crest cell. NCC-melb4 cells provide a model to study the specific stage of differentiation and proliferation of melanocytes. They also express KIT as a melanoblast marker. Using the NCC-melb4 cell line, we investigated the effect of transforming growth factor beta1 on the differentiation and proliferation of immature melanocyte precursors. Immunohistochemically, NCC-melb4 cells showed transforming growth factor beta1 expression. The anti-transforming growth factor beta1 antibody inhibited the cell growth, and downregulated the KIT protein and mRNA expression. To investigate further the activation of autocrine transforming growth factor beta1, NCC-melb4 cells were incubated in nonexogenous transforming growth factor beta1 culture medium. KIT protein decreased with anti-transforming growth factor beta1 antibody concentration in a concentration-dependent manner. We concluded that in NCC-melb4 cells, transforming growth factor beta1 promotes melanocyte precursor proliferation in autocrine and/or paracrine regulation. We further investigated the influence of transforming growth factor beta1 in vitro using a neural crest cell primary culture system from wild-type mice. Anti-transforming growth factor beta1 antibody decreased the number of KIT positive neural crest cell. In addition, the anti-transforming growth factor beta1 antibody supplied within the wild-type neural crest explants abolished the growth of the neural

  7. Dahlbeck and Pure Ontology

    Science.gov (United States)

    Mackenzie, Jim

    2016-01-01

    This article responds to Johan Dahlbeck's "Towards a pure ontology: Children's bodies and morality" ["Educational Philosophy and Theory," vol. 46 (1), 2014, pp. 8-23 (EJ1026561)]. His arguments from Nietzsche and Spinoza do not carry the weight he supposes, and the conclusions he draws from them about pedagogy would be…

  8. Dahlbeck and Pure Ontology

    Science.gov (United States)

    Mackenzie, Jim

    2016-01-01

    This article responds to Johan Dahlbeck's "Towards a pure ontology: Children's bodies and morality" ["Educational Philosophy and Theory," vol. 46 (1), 2014, pp. 8-23 (EJ1026561)]. His arguments from Nietzsche and Spinoza do not carry the weight he supposes, and the conclusions he draws from them about pedagogy would be…

  9. INGESTIVE BEHAVIOR OF GOATS IN RYEGRASS AND BLACK OAT PASTURES IN PURE OR MIXTURE CULTURE COMPORTAMENTO INGESTIVO DE CAPRINOS EM PASTAGEM DE AZEVÉM E AVEIA PRETA EM CULTIVO PURO E CONSORCIADO

    Directory of Open Access Journals (Sweden)

    Alda Lúcia Gomes Monteiro

    2009-07-01

    Full Text Available The experiment was realized in Campo Largo, PR, where the ingestive behavior of goats was evaluated under ryegrass (Lolium multiflorum Lam. and black oat (Avena strigosa Schreb pastures in pure or mixture culture, in the period of 04/07/2004 to 05/08/2004. The grasses were applied in poles of 630 m² each, and the experimental design was placed in randomized blocks with three treatments and three repetitions. Twelve female goats were distributed in three experimental poles with four goats each for grazing evaluations. Previously to the evaluations of the animals the measurements of the pasture were obtained, which included height, total mass of forage and of the compounds leaf and steam. The goats were evaluated by preference and ingestion rate. The averages of pastures height was higher (p>0.05 in ryegrass and mixture, and in other pastures evaluations ryegrass was superior (p<0.05 to the others treatments. The grazing time of goats in ryegrass and black oat was superior (p<0.05 to the mixture. The bite rate per minute was higher (p<0.05 in black oat. The goats demonstrated preference for ryegrass and black oat in pure culture.
     
    KEY WORDS: Avena strigosa Schreb, bite, goats, Lolium multiflorum Lam, preference. The experiment was realized in Campo Largo – PR, where the ingestive behavior of goats was evaluated under ryegrass (Lolium multiflorum Lam. and black oat (Avena strigosa Schreb pastures in pure or mixture culture, in the period of 04/07/2004 to 05/08/2004. The grasses were applied in poles of 630 m² each, and the experimental design was placed in randomized blocks with three treatments and three repetitions. Twelve female goats were distributed in three experimental poles with four goats each for grazing evaluations. Previously to the evaluations of the animals the measurements of the pasture were obtained, which included height, total mass of forage and of the compounds leaf and steam. The goats were evaluated by

  10. Relationships between melanocytes, mechanical properties and extracellular matrix composition in mouse heart valves.

    Science.gov (United States)

    Carneiro, Flavia; Kruithof, Boudewijn Pt; Balani, Kanthesh; Agarwal, Arvind; Gaussin, Vinciane; Kos, Lidia

    2015-01-01

    Heart valves are complex structures composed of organized layers of extracellular matrix, and interstitial and overlying endothelial cells. In this article, we present the specific localization of a population of melanocytes within the murine heart valves at ages important for their post-natal development. In all stages analyzed in our study, melanocytes were found in high numbers populating the atrial aspect of the tricuspid and mitral leaflets. The pulmonary valve did not present melanocytes. To characterize a putative role for the valve melanocytes, the dynamic nanomechanical properties of tricuspid leaftets containing large numbers or no melanocytes were measured. The stiffness coefficient of hyperpigmented leaflets was higher (11.5 GPa) than the ones from wild-type (7.5 GPa) and hypopigmented (5.5 GPa) leaflets. These results suggest that melanocytes may contribute to the mechanical properties of the heart valves. The arrangement of extracellular matrix molecules such as Collagen I and Versican B is responsible for the mechanical characteristics of the leaflets. Melanocytes were found to reside primarily in areas of Versican B expression. The patterns of expression of Collagen I and Versican B were not, however, disrupted in hyper or hypopigmented leaflets. Melanocytes may affect other extracellular matrix molecules to alter the valves' microenvironment.

  11. Constitutive gray hair in mice induced by melanocyte-specific deletion of c-Myc

    Science.gov (United States)

    Pshenichnaya, Irina; Schouwey, Karine; Armaro, Marzia; Larue, Lionel; Knoepfler, Paul S.; Eisenman, Robert N.; Trumpp, Andreas; Delmas, Véronique; Beermann, Friedrich

    2012-01-01

    Summary c-Myc is involved in the control of diverse cellular processes and implicated in the maintenance of different tissues including the neural crest. Here, we report that c-Myc is particularly important for pigment cell development and homeostasis. Targeting c-Myc specifically in the melanocyte lineage using the floxed allele of c-Myc and Tyr::Cre transgenic mice results in a congenital gray hair phenotype. The gray coat color is associated with a reduced number of functional melanocytes in the hair bulb and melanocyte stem cells in the hair bulge. Importantly, the gray phenotype does not progress with time, suggesting that maintenance of the melanocyte through the hair cycle does not involve c-Myc function. In embryos, at E13.5, c-Myc-deficient melanocyte precursors are affected in proliferation in concordance with a reduction in numbers, showing that c-Myc is required for the proper melanocyte development. Interestingly, melanocytes from c-Myc-deficient mice display elevated levels of the c-Myc paralog N-Myc. Double deletion of c-Myc and N-Myc results in nearly complete loss of the residual pigmentation, indicating that N-Myc is capable of compensating for c-Myc loss of function in melanocytes. PMID:22420299

  12. Giant Congenital Melanocytic Nevus with Developmental Dysplasia of Bilateral Hip: A Rare Association

    OpenAIRE

    Sutsungkokla Imchen; Sangita Ghosh; Surabhi Dayal; Nisha Marwah; Nidhi Jindal; Shikha Sangal

    2013-01-01

    Giant congenital melanocytic nevi are rare congenital disfiguring benign neoplasms with a risk of transformation to malignant melanoma. They often present with various extra-cutaneous features. Here, we describe a case of giant melanocytic nevus with developmental dysplasia of bilateral hip, a novel association.

  13. Giant congenital melanocytic nevus with developmental dysplasia of bilateral hip: a rare association.

    Science.gov (United States)

    Imchen, Sutsungkokla; Ghosh, Sangita; Dayal, Surabhi; Marwah, Nisha; Jindal, Nidhi; Sangal, Shikha

    2013-11-01

    Giant congenital melanocytic nevi are rare congenital disfiguring benign neoplasms with a risk of transformation to malignant melanoma. They often present with various extra-cutaneous features. Here, we describe a case of giant melanocytic nevus with developmental dysplasia of bilateral hip, a novel association.

  14. Coexistence of congenital giant melanocytic nevus of the scalp with cranial defect, poliosis, and hair loss.

    Science.gov (United States)

    Lee, Woo J; Lee, Sang M; Won, Chong H; Chang, Sung E; Lee, Mi W; Choi, Jee H; Moon, Kee C

    2013-01-01

    Congenital melanocytic nevi (CMN) are pigmented lesions presenting on the skin in approximately 1% of all newborns at or shortly after birth. CMN have been described as being associated with several anomalies, including cranial bone hypertrophy, scoliosis, and spina bifida. This is the first report to describe a giant congenital melanocytic nevus on the scalp associated with cranial involvement, poliosis, and alopecia.

  15. Spindle cell melanocytic lesions: part II--an approach to intradermal proliferations and horizontally oriented lesions.

    Science.gov (United States)

    Sade, Shachar; Al Habeeb, Ayman; Ghazarian, Danny

    2010-05-01

    Melanocytic lesions show great morphological diversity in their architecture and the cytomorphological appearance of their composite cells. Whereas functional melanocytes show a dendritic cytomorphology and territorial isolation, lesional nevomelanocytes and melanoma cells typically show epithelioid, spindled or mixed cytomorphologies, and a range of architectural arrangements. Spindling is common to melanocytic lesions, and may either be a characteristic feature or a divergent appearance. The presence of spindle cells may mask the melanocytic nature of a lesion, and is often disconcerting, either due to its infrequent appearance in a particular lesion or its interpretation as a dedifferentiated phenotype. Spindle cell melanocytic lesions follow the full spectrum of potential biological outcomes, and difficulty may be experienced judging the nature of a lesion due to a lack of consistently reliable features to predict biological behaviour. Over time, recognition of numerous histomorphological features that may portend a more aggressive lesion have been identified; however, the translation of these features into a diagnostic entity requires a gestalt approach. Although most spindle cell melanocytic lesions may reliably be resolved through this standard approach, problem areas do exist for the surgical pathologist or dermatopathologist. With this review (part II of II), we complete our discussion of spindle cell melanocytic lesions, in order to: (1) model a systematic approach to such lesions; and (2) provide familiarity with those melanocytic lesions which either typically or occasionally display a spindled cytomorphology.

  16. Repression of genes involved in melanocyte differentiation in uveal melanoma

    Science.gov (United States)

    Bergeron, Marjorie-Allison; Champagne, Sophie; Gaudreault, Manon; Deschambeault, Alexandre

    2012-01-01

    Purpose Uveal melanoma (UM) has been the subject of intense interest due to its distinctive metastatic pattern, which involves hematogenous dissemination of cancerous cells toward the liver in 50% of patients. To search for new UM prognostic markers, the Suppressive Subtractive Hybridization (SSH) technique was used to isolate genes that are differentially expressed between UM primary tumors and normal uveal melanocytes (UVM). Methods A subtracted cDNA library was prepared using cDNA from uncultured UM primary tumors and UVM. The expression level of selected genes was further validated by cDNA microarray, semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), and immunofluorescence analyses. Results One hundred-fifteen genes were identified using the SSH technique. Microarray analyses comparing the gene expression profiles of UM primary tumors to UVM validated a significant differential expression for 48% of these genes. The expression pattern of selected genes was then analyzed by semi-quantitative RT–PCR and was found to be consistent with the SSH and cDNA microarray findings. A down-regulation of genes associated with melanocyte differentiation was confirmed in UM primary tumors. Presence of undifferentiated cells in the UM was demonstrated by the expression of stem cell markers ATP-binding cassette sub-family G member 2 (ABCG2) and octamer-binding protein 4 (OCT4). Conclusions We demonstrated that the SSH technique is efficient to detect differentially expressed genes between UM and UVM. The genes identified in this study represent valuable candidates for further functional analysis in UM and should be informative in studying the biology of this tumor. In addition, deregulation of the melanocyte differentiation pathway revealed the presence of UM cells exhibiting a stem cell-like phenotype. PMID:22815634

  17. Purely Functional Structured Programming

    OpenAIRE

    Obua, Steven

    2010-01-01

    The idea of functional programming has played a big role in shaping today's landscape of mainstream programming languages. Another concept that dominates the current programming style is Dijkstra's structured programming. Both concepts have been successfully married, for example in the programming language Scala. This paper proposes how the same can be achieved for structured programming and PURELY functional programming via the notion of LINEAR SCOPE. One advantage of this proposal is that m...

  18. Purely Cortical Anaplastic Ependymoma

    Directory of Open Access Journals (Sweden)

    Flávio Ramalho Romero

    2012-01-01

    Full Text Available Ependymomas are glial tumors derived from ependymal cells lining the ventricles and the central canal of the spinal cord. It may occur outside the ventricular structures, representing the extraventicular form, or without any relationship of ventricular system, called ectopic ependymona. Less than fifteen cases of ectopic ependymomas were reported and less than five were anaplastic. We report a rare case of pure cortical ectopic anaplastic ependymoma.

  19. Oral Congenital Melanocytic Nevus: A Rare Case Report and Review of the Literature.

    Science.gov (United States)

    Marangon Júnior, Helvécio; Souza, Paulo Eduardo Alencar; Soares, Rodrigo Villamarim; de Andrade, Bruno Augusto Benevenuto; de Almeida, Oslei Paes; Horta, Martinho Campolina Rebello

    2015-12-01

    Melanocytic nevi are congenital or acquired benign proliferations of cells of melanocytic origin. Oral congenital melanocytic nevi are rare, and only a few cases have been reported in the literature. The purpose of this study is to present the clinical, histological and immunohistochemical features of an oral congenital melanocytic nevus in a 16-year-old female with an 11-year follow-up and to review the pertinent literature. The reported case is the fifth well-documented case report of oral congenital melanocytic nevus in the English literature and the first with a long period of follow-up, thereby making it an important contribution to the knowledge regarding this uncommon oral mucosa lesion.

  20. Purely tetrahedral quadruple systems

    Institute of Scientific and Technical Information of China (English)

    JI Lijun

    2006-01-01

    An oriented tetrahedron is a set of four vertices and four cyclic triples with the property that any ordered pair of vertices is contained in exactly one of the cyclic triples. A tetrahedral quadruple system of order n (briefly TQS(n)) is a pair (X,B), where X is an nelement set and B is a set of oriented tetrahedra such that every cyclic triple on X is contained in a unique member of B. A TQS(n) (X, B) is pure if there do not exist two oriented tetrahedra with the same vertex set. In this paper, we show that there is a pure TQS(n) if and only if n≡2,4(mod 6),n>4,or n≡1,5(mod 12). One corollary is that there is a simple two-fold quadruple system of order n if and only if n≡2,4 (mod 6) and n>4, or n≡1, 5 (mod 12).Another corollary is that there is an overlarge set of pure Mendelsohn triple systems of order n for n≡1,3(mod 6),n>3, or n≡0,4 (mod 12).

  1. Deep penetrating nevus: A distinct variant of melanocytic nevus

    Directory of Open Access Journals (Sweden)

    Aparna Gupta

    2011-01-01

    Full Text Available Deep penetrating nevus (DPN is a variant of melanocytic nevus which goes unrecognized due to its relative rarity and may be misinterpreted as malignant melanoma. It commonly presents in young adults as a dark pigmented lesion on the face, neck, or shoulder. A 60-year-old lady presented with a mole over the left arm of 8 years duration. A biopsy of the lesion was performed under the clinical impression of a compound nevus with suspicion of malignancy. Based on the histologic features, a diagnosis of DPN was put forward.

  2. Size Functions for the Morphological Analysis of Melanocytic Lesions

    Science.gov (United States)

    Ferri, Massimo; Stanganelli, Ignazio

    2010-01-01

    Size Functions and Support Vector Machines are used to implement a new automatic classifier of melanocytic lesions. This is mainly based on a qualitative assessment of asymmetry, performed by halving images by several lines through the center of mass, and comparing the two halves in terms of color, mass distribution, and boundary. The program is used, at clinical level, with two thresholds, so that comparison of the two outputs produces a report of low-middle-high risk. Experimental results on 977 images, with cross-validation, are reported. PMID:20300598

  3. Size Functions for the Morphological Analysis of Melanocytic Lesions

    Directory of Open Access Journals (Sweden)

    Massimo Ferri

    2010-01-01

    Full Text Available Size Functions and Support Vector Machines are used to implement a new automatic classifier of melanocytic lesions. This is mainly based on a qualitative assessment of asymmetry, performed by halving images by several lines through the center of mass, and comparing the two halves in terms of color, mass distribution, and boundary. The program is used, at clinical level, with two thresholds, so that comparison of the two outputs produces a report of low-middle-high risk. Experimental results on 977 images, with cross-validation, are reported.

  4. 成纤维细胞生长因子对黑素细胞生长的影响%Effects of basic fibroblast growth factor (bFGF) on the growth of melanocytes

    Institute of Scientific and Technical Information of China (English)

    丁克云; 赵志国

    2010-01-01

    Objective To study the effects of bFGF on the proliferation of cultured human melanocytes, and to seek a quick method for in vitro culture of human melanocytes. Methods Melanocytes were isolated from human foreskin, and divided into two parts to be cultured with or without the presence of bFGF (0.3 μg/L). Second-passage melanocytes were identified with immunochemical stain. The growth of melanocytes was observed every 3 days for 12 days. Third-passage melanocytes were treated with various concentrations (0.3 - 2.1 μg/L) of bFGF for 72 hours followed by the detection of proliferation of and trosinase activity in melanocytes. Results Human melanocytes were obtained from primary culture in medium containing certain concentrations of bFGF, which were identified with immunohistochemical stain. The morphology of cultured melanocytes varied with growth stage of cells. The bFGF-treated melanocytes appeared to grow more rapidly than untreated melanocytes. Further more, a significant increase was observed in the proliferation rate of melanocytes treated with bFGF of 0.3 and 0.6 μg/L (P<0.05 or 0.01 ) and tyrosinase activity in melanocytes treated with bFGF of 1.5 and 1.8 μg/L (P < 0.05 or 0.01 ) in comparison with the untreated melanocytes.Conclusions The addition of certain concentrations of bFGF to defined medium can benefit the primary culture of melanocytes and make it possible to get large quantities of purified melanocytes with high viability in short periods. Certain concentrations of bFGF can up-regulate the proliferation of and tyrosinase activity in melanocytes.%目的 探讨成纤维细胞生长因子(bFGF)对体外培养正常人黑素细胞增殖的影响,寻找快速体外培养正常人黑素细胞的方法.方法 分别用普通的合成培养基与添加一定浓度bFGF的合成培养基原代培养正常人黑素细胞,观察两种条件下细胞生长情况,并对培养的黑素细胞进行鉴定,对不同生长期黑素细胞的形

  5. Increased endogenous DNA oxidation correlates to increased iron levels in melanocytes relative to keratinocytes.

    Science.gov (United States)

    Pelle, Edward; Huang, Xi; Zhang, Qi; Pernodet, Nadine; Yarosh, Daniel B; Frenkel, Krystyna

    2014-01-01

    The endogenous oxidative state of normal human epidermal melanocytes was investigated and compared to normal human epidermal keratinocytes (NHEKs) in order to gain new insight into melanocyte biology. Previously, we showed that NHEKs contain higher levels of hydrogen peroxide (H2O2) than melanocytes and that it can migrate from NHEKs to melanocytes by passive permeation. Nevertheless, despite lower concentrations of H2O2, we now report higher levels of oxidative DNA in melanocytes as indicated by increased levels of 8-oxo-2'-deoxyguanosine (8-oxo-dG): 4.49 (±0.55 SEM) 8-oxo-dG/10(6) dG compared to 1.49 (±0.11 SEM) 8-oxo-dG/10(6) dG for NHEKs. An antioxidant biomarker, glutathione (GSH), was also lower in melanocytes (3.14 nmoles (±0.15 SEM)/cell) in comparison to NHEKs (5.98 nmoles (±0.33 SEM)/cell). Intriguingly, cellular bioavailable iron as measured in ferritin was found to be nearly fourfold higher in melanocytes than in NHEKs. Further, ferritin levels in melanocytes were also higher than in hepatocarcinoma cells, an iron-rich cell, and it indicates that higher relative iron levels may be characteristic of melanocytes. To account for the increased oxidative DNA and lower GSH and H2O2 levels that we observe, we propose that iron may contribute to higher levels of oxidation by reacting with H2O2 through a Fenton reaction leading to the generation of DNA-reactive hydroxyl radicals. In conclusion, our data support the concept of elevated oxidation and high iron levels as normal parameters of melanocytic activity. We present new evidence that may contribute to our understanding of the melanogenic process and lead to the development of new skin care products.

  6. Pure and Public, Popular and personal

    DEFF Research Database (Denmark)

    Eriksson, Birgit

    2013-01-01

    In the article I reexamine the traditional aesthetical and political critiques of popular culture and reevaluate the social and communicative potential of bestselling cultural artifacts such as highly popular television series. First, I sketch the alleged aesthetic and social problems of popular...... and the exclusions of the public sphere. I argue that the ideals of a pure aesthetic and a public sphere neglect issues that are crucial to the type of commonality at stake in popular cultural artifacts: personal issues, social conflicts, and what is pleasurable to the senses or has to do with emotions. Third, I...

  7. PURE DRIVE GT

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    在2004年奥林匹克赛事中,中国的李婷,孙甜甜取得了中国网球第一个金牌一女子双打冠军。忘记不了当时李婷挥动着她的BABOLAT(百保力)网拍Pure Drive Zylon 360°激动地拥抱着孙甜甜吵闹着,幸福地哭着的情景。

  8. Pure de Sitter Supergravity

    CERN Document Server

    Bergshoeff, Eric A; Kallosh, Renata; Van Proeyen, Antoine

    2015-01-01

    Using superconformal methods we derive an explicit de Sitter supergravity action invariant under spontaneously broken local ${\\cal N}=1$ supersymmetry. The supergravity multiplet interacts with a nilpotent goldstino multiplet. We present a complete locally supersymmetric action including the graviton and the fermionic fields, gravitino and goldstino, no scalars. In the global limit when supergravity multiplet decouples, our action reproduces the Volkov-Akulov theory. In the unitary gauge where goldstino vanishes we recover pure supergravity with the positive cosmological constant. The classical equations of motion, with all fermions vanishing, have a maximally symmetric solution: de Sitter space.

  9. SZ95 sebocytes induce epidermal melanocyte dendricity and proliferation in vitro.

    Science.gov (United States)

    Abdel-Naser, Mohamed Badawy; Seltmann, Holger; Zouboulis, Christos C

    2012-05-01

    The regulatory effects of sebocytes on melanocytes (HMel) are unknown. In this study, SZ95 sebocytes co-cultured with HMel, whether in direct cell contact or with SZ95 sebocytes in inserts, resulted in epidermal HMel flattening with increase in surface area and multiple small dendrites formation. Only in high Ca(2+) level and direct cell contact co-culture, the HMel dendrites were remarkably long and preferentially targeted and attached to SZ95 sebocytes. Likewise, only high Ca(2+) SZ95 sebocyte conditioned medium stimulated HMel proliferation in a time-dependent manner at days 9 (142.9%, P < 0.01) and 12 (179.2%, P < 0.0001) of incubation when compared with day 0. In contrast, melanin contents significantly decreased on incubation with high Ca(2+) SZ95 sebocytes in comparison with low Ca(2+) SZ95 sebocytes at days 6 (P < 0.01) and 9 (P < 0.05) of incubation. These results denote that sebocytes also modulate HMel functions and may contribute to skin colour in sebaceous glands-rich body regions.

  10. A Micro-RNA Connection in BRafV600E-Mediated Premature Senescence of Human Melanocytes

    Directory of Open Access Journals (Sweden)

    Gang Ren

    2012-01-01

    Full Text Available Recent high-throughput-sequencing of the cancer genome has identified oncogenic mutations in BRaf genetic locus as one of the critical events in melanomagenesis. In normal cells, the activity of BRaf is tightly regulated. Gain-of-function mutations like those identified in melanoma frequently lead to enhanced cell-survival and unrestrained growth. The activating mutation of BRaf will also induce the cells to senesce. However, the mechanism by which the oncogenic BRaf induces the senescent barrier remains poorly defined. microRNAs have regulatory functions toward the expression of genes that are important in carcinogenesis. Here we show that expression of several microRNAs is altered when the oncogenic version of BRaf is introduced in cultured primary melanocytes and these cells undergo premature cellular senescence. These include eight microRNAs whose expression rates are significantly stimulated and three that are repressed. While most of the induced microRNAs have documented negative effects on cell cycle progression, one of the repressed microRNAs has proven oncogenic functions. Ectopic expression of some of these induced microRNAs increased the expression of senescence markers and induced growth arrest and senescence in primary melanocytes. Taken together, our results suggest that the change in microRNA expression rates may play a vital role in senescence induced by the oncogenic BRaf.

  11. Protective Effect of HemoHIM on Epidermal Melanocytes in Ultraviolet-B irradiated Mice

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hae June [Korea Institute of Radiological and Medical Science, Seoul (Korea, Republic of); Kim, Jong Choon; Moon, Chang Jong; Kim, Sung Ho [Chonnam National University, Gwangju (Korea, Republic of); Jung, U Hee; Park, Hae Ran; Jo, Sung Kee [Jeongeup Campus of Korea Atomic Energy Research Institute, Jeongeup (Korea, Republic of); Jang, Jong Sik; Kim, Tae Hwan [Kyungpook National University, Daegu (Korea, Republic of)

    2011-06-15

    We induced the activation of melanocytes in the epidermis of C57BL/6 mice by ultraviolet-B (UV-B) irradiation, and observed the effect of an herbal preparation (HemoHIM, HH) on the formation, and decrease of UV-B-induced epidermal melanocytes. C57BL/6 mice were irradiated by UV-B 80 mJ:cm{sup -2} (0.5 mW:sec{sup -1}) daily for 7 days, and HH was intraperitoneally, orally or topically applied pre- or post-irradiation. For the estimation of change of epidermal melanocytes, light microscopic observation with dihydroxyphenylalanine (DOPA) stain was performed. Split epidermal sheets prepared from the ear of untreated mice exhibited 13∼15 melanocytes:mm{sup -2}, and one week after UV irradiation, the applied areas showed an increased number of strongly DOPA-positive melanocytes with stout dendrites. But intraperitoneal, oral or topical treatment with HH before each irradiation interrupted UV-B-induced pigmentation and resulted in a marked reduction in the number of epidermal melanocytes as compared to the number found in UV-B-irradiated, untreated control skin. The number and size of DOPA-positive epidermal melanocytes were also significantly decreased in intraperitoneally injected or topically applicated group after irradiation with HH at 3rd and 6th weeks after irradiation. The present study suggests the HH as inhibitor of UV-B-induced pigmentation, and depigmenting agent.

  12. RICTOR involvement in the PI3K/AKT pathway regulation in melanocytes and melanoma.

    Science.gov (United States)

    Laugier, Florence; Finet-Benyair, Adeline; André, Jocelyne; Rachakonda, P Sivaramakrishna; Kumar, Rajiv; Bensussan, Armand; Dumaz, Nicolas

    2015-09-29

    Several studies have highlighted the importance of the PI3K pathway in melanocytes and its frequent over-activation in melanoma. However, little is known about regulation of the PI3K pathway in melanocytic cells. We showed that normal human melanocytes are less sensitive to selective PI3K or mTOR inhibitors than to dual PI3K/mTOR inhibitors. The resistance to PI3K inhibitor was due to a rapid AKT reactivation limiting the inhibitor effect on proliferation. Reactivation of AKT was linked to a feedback mechanism involving the mTORC2 complex and in particular its scaffold protein RICTOR. RICTOR overexpression in melanocytes disrupted the negative feedback, activated the AKT pathway and stimulated clonogenicity highlighting the importance of this feedback to restrict melanocyte proliferation. We found that the RICTOR locus is frequently amplified and overexpressed in melanoma and that RICTOR over-expression in NRAS-transformed melanocytes stimulates their clonogenicity, demonstrating that RICTOR amplification can cooperate with NRAS mutation to stimulate melanoma proliferation. These results show that RICTOR plays a central role in PI3K pathway negative feedback in melanocytes and that its deregulation could be involved in melanoma development.

  13. Race Does Not Predict Melanocyte Heterogeneous Responses to Dermal Fibroblast-Derived Mediators.

    Directory of Open Access Journals (Sweden)

    Pornthep Sirimahachaiyakul

    Full Text Available Abnormal pigmentation following cutaneous injury causes significant patient distress and represents a barrier to recovery. Wound depth and patient characteristics influence scar pigmentation. However, we know little about the pathophysiology leading to hyperpigmentation in healed shallow wounds and hypopigmentation in deep dermal wound scars. We sought to determine whether dermal fibroblast signaling influences melanocyte responses.Epidermal melanocytes from three Caucasians and three African-Americans were genotyped for single nucleotide polymorphisms (SNPs across the entire genome. Melanocyte genetic profiles were determined using principal component analysis. We assessed melanocyte phenotype and gene expression in response to dermal fibroblast-conditioned medium and determined potential mesenchymal mediators by proteome profiling the fibroblast-conditioned medium.Six melanocyte samples demonstrated significant variability in phenotype and gene expression at baseline and in response to fibroblast-conditioned medium. Genetic profiling for SNPs in receptors for 13 identified soluble fibroblast-secreted mediators demonstrated considerable heterogeneity, potentially explaining the variable melanocyte responses to fibroblast-conditioned medium.Our data suggest that melanocytes respond to dermal fibroblast-derived mediators independent of keratinocytes and raise the possibility that mesenchymal-epidermal interactions influence skin pigmentation during cutaneous scarring.

  14. Preferential secretion of inducible HSP70 by vitiligo melanocytes under stress.

    Science.gov (United States)

    Mosenson, Jeffrey A; Flood, Kelsey; Klarquist, Jared; Eby, Jonathan M; Koshoffer, Amy; Boissy, Raymond E; Overbeck, Andreas; Tung, Rebecca C; Le Poole, I Caroline

    2014-03-01

    Inducible HSP70 (HSP70i) chaperones peptides from stressed cells, protecting them from apoptosis. Upon extracellular release, HSP70i serves an adjuvant function, enhancing immune responses to bound peptides. We questioned whether HSP70i differentially protects control and vitiligo melanocytes from stress and subsequent immune responses. We compared expression of HSP70i in skin samples, evaluated the viability of primary vitiligo and control melanocytes exposed to bleaching phenols, and measured secreted HSP70i. We determined whether HSP70i traffics to melanosomes to contact immunogenic proteins by cell fractionation, western blotting, electron microscopy, and confocal microscopy. Viability of vitiligo and control melanocytes was equally affected under stress. However, vitiligo melanocytes secreted increased amounts of HSP70i in response to MBEH, corroborating with aberrant HSP70i expression in patient skin. Intracellular HSP70i colocalized with melanosomes, and more so in response to MBEH in vitiligo melanocytes. Thus, whereas either agent is cytotoxic to melanocytes, MBEH preferentially induces immune responses to melanocytes.

  15. Dct::lacZ ES cells: a novel cellular model to study melanocyte determination and differentiation.

    Science.gov (United States)

    Pla, Patrick; Solov'eva, Olga; Moore, Robert; Alberti, Christophe; Kunisada, Takahiro; Larue, Lionel

    2004-04-01

    Embryonic stem (ES) cells differentiate into various cell lineages in vitro. A procedure was previously designed to promote the differentiation of ES cells towards the melanocyte lineage and to obtain large and reproducible amounts of melanocytes. To elucidate the main events that lead to the development of melanocytes in vitro, we used transgenic Dct::lacZ mouse blastocysts to establish ES cell lines expressing the lacZ reporter gene under the control of the Dct promoter. Dct, a melanoblast marker, is expressed just after melanoblast determination in vivo. We evaluated the importance of recruitment, proliferation and differentiation during melanocyte ontogeny after the in vitro differentiation of Dct::lacZ ES cells into melanocytes. We showed that bFGF and cholera toxin induce precocious melanoblast determination, associated with early melanocyte differentiation. Edn3 induced melanoblast proliferation and long-term melanoblast recruitment, but not precocious determination. The lack of basic Fibroblast Growth Factor (bFGF) and cholera toxin can be partially compensated by Edn3. Thus, Dct::lacZ ES cells can be used as a model to study determination, proliferation and differentiation in the melanocyte lineage in vitro.

  16. Differential PAX3 functions in normal skin melanocytes and melanoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Medic, Sandra [School of Exercise, Biomedical and Health Sciences, Edith Cowan University, Perth, WA (Australia); Rizos, Helen [Westmead Institute for Cancer Research and Melanoma Institute of Australia, University of Sydney at Westmead Millennium Institute, Westmead, NSW (Australia); Ziman, Mel, E-mail: m.ziman@ecu.edu.au [School of Exercise, Biomedical and Health Sciences, Edith Cowan University, Perth, WA (Australia); School of Pathology and Laboratory Medicine, University of Western Australia, Perth, WA (Australia)

    2011-08-12

    Highlights: {yields} PAX3 retains embryonic roles in adult melanocytes and melanoma cells. {yields} Promotes 'stem' cell-like phenotype via NES and SOX9 in both cells types. {yields} Regulates melanoma and melanocyte migration through MCAM and CSPG4. {yields} PAX3 regulates melanoma but not melanocyte proliferation via TPD52. {yields} Regulates melanoma cell (but not melanocyte) survival via BCL2L1 and PTEN. -- Abstract: The PAX3 transcription factor is the key regulator of melanocyte development during embryogenesis and is also frequently found in melanoma cells. While PAX3 is known to regulate melanocyte differentiation, survival, proliferation and migration during development, it is not clear if its function is maintained in adult melanocytes and melanoma cells. To clarify this we have assessed which genes are targeted by PAX3 in these cells. We show here that similar to its roles in development, PAX3 regulates complex differentiation networks in both melanoma cells and melanocytes, in order to maintain cells as 'stem' cell-like (via NES and SOX9). We show also that mediators of migration (MCAM and CSPG4) are common to both cell types but more so in melanoma cells. By contrast, PAX3-mediated regulation of melanoma cell proliferation (through TPD52) and survival (via BCL2L1 and PTEN) differs from that in melanocytes. These results suggest that by controlling cell proliferation, survival and migration as well as maintaining a less differentiated 'stem' cell like phenotype, PAX3 may contribute to melanoma development and progression.

  17. In ovo gene manipulation of melanocytes and their adjacent keratinocytes during skin pigmentation of chicken embryos.

    Science.gov (United States)

    Murai, Hidetaka; Tadokoro, Ryosuke; Sakai, Ken-Ichiro; Takahashi, Yoshiko

    2015-04-01

    During skin pigmentation in avians and mammalians, melanin is synthesized in the melanocytes, and subsequently transferred to adjacently located keratinocytes, leading to a wide coverage of the body surface by melanin-containing cells. The behavior of melanocytes is influenced by keratinocytes shown mostly by in vitro studies. However, it has poorly been investigated how such intercellular cross-talk is regulated in vivo because of a lack of suitable experimental models. Using chicken embryos, we developed a method that enables in vivo gene manipulations of melanocytes and keratinocytes, where these cells are separately labeled by different genes. Two types of gene transfer techniques were combined: one was a retrovirus-mediated gene infection into the skin/keratinocytes, and the other was the in ovo DNA electroporation into neural crest cells, the origin of melanocytes. Since the Replication-Competent Avian sarcoma-leukosis virus long terminal repeat with Splice acceptor (RCAS) infection was available only for the White leghorn strain showing little pigmentation, melanocytes prepared from the Hypeco nera (pigmented) were back-transplanted into embryos of White leghorn. Prior to the transplantation, enhanced green fluorescent protein (EGFP)(+) Neo(r+) -electroporated melanocytes from Hypeco nera were selectively grown in G418-supplemented medium. In the skin of recipient White leghorn embryos infected with RCAS-mOrange, mOrange(+) keratinocytes and transplanted EGFP(+) melanocytes were frequently juxtaposed each other. High-resolution confocal microscopy also revealed that transplanted melanocytes exhibited normal behaviors regarding distribution patterns of melanocytes, dendrite morphology, and melanosome transfer. The method described in this study will serve as a useful tool to understand the mechanisms underlying intercellular regulations during skin pigmentation in vivo.

  18. Dual roles of lineage restricted transcription factors: the case of MITF in melanocytes.

    Science.gov (United States)

    Levy, Carmit; Fisher, David E

    2011-01-01

    Microphthalmia-associated Transcription Factor, MITF, is a master regulator of melanocyte development, differentiation, migration, and survival.(1) A broad collection of studies have indicated that MITF directly regulates the transcription of genes involved in pigmentation, which are selective to the melanocyte lineage. In addition, MITF controls expression of genes which are expressed in multiple cell lineages, and may also play differential roles in activating vs. maintaining gene expression patterns. In this Point of View article, we discuss lineage restricted transcription factor activation of both tissue-specific and ubiquitously expressed genes using melanocytes and MITF as a model system that may eventually provide insights into such processes in multiple cell lineages.

  19. Stereological estimation of nuclear volume in benign melanocytic lesions and cutaneous malignant melanomas

    DEFF Research Database (Denmark)

    Sørensen, Flemming Brandt

    1989-01-01

    V in melanocytic cutaneous tumors and to compare these with estimates of nuclear volume fraction and with traditional two-dimensional morphometric estimates of nuclear profile area, nuclear density, and mitotic index. Routinely processed, paraffin embedded tissue specimens from 47 malignant melanomas and 76...... noninvasive melanocytic cutaneous tumors were investigated retrospectively. vV clearly distinguished between noninvasive (average vV = 122 microns 3) and invasive lesions (average vV = 246 microns 3). Most of the patients with malignant melanomas showing an overlap of nuclear vV with benign lesions had...... estimator for distinguishing between melanocytic cutaneous tumors showing different biological behavior, well-suited for objective malignancy grading....

  20. Mutations in c10orf11, a melanocyte-differentiation gene, cause autosomal-recessive albinism

    DEFF Research Database (Denmark)

    Grønskov, Karen; Dooley, Christopher M; Østergaard, Elsebet

    2013-01-01

    in an individual originating from Lithuania. Immunohistochemistry showed localization of C10orf11 in melanoblasts and melanocytes in human fetal tissue, but no localization was seen in retinal pigment epithelial cells. Knockdown of the zebrafish (Danio rerio) homolog with the use of morpholinos resulted...... in substantially decreased pigmentation and a reduction of the apparent number of pigmented melanocytes. The morphant phenotype was rescued by wild-type C10orf11, but not by mutant C10orf11. In conclusion, we have identified a melanocyte-differentiation gene, C10orf11, which when mutated causes autosomal...

  1. Pure laparoscopic augmentation ileocystoplasty.

    Science.gov (United States)

    Rebouças, Rafael B; Monteiro, Rodrigo C; Souza, Thiago N S de; Aragão, Augusto J de; Burity, Camila R T; Nóbrega, Júlio C de A; Oliveira, Natália S C de; Abrantes, Ramon B; Dantas Júnior, Luiz B; Cartaxo Filho, Ricardo; Negromonte, Gustavo R P; Sampaio, Rafael da C R; Britto, Cesar A

    2014-01-01

    Guillain-Barre syndrome is an acute neuropathy that rarely compromises bladder function. Conservative management including clean intermittent catheterization and pharmacotherapy is the primary approach for hypocompliant contracted bladder. Surgical treatment may be used in refractory cases to improve bladder compliance and capacity in order to protect the upper urinary tract. We describe a case of pure laparoscopic augmentation ileocystoplasty in a patient affected by Guillain-Barre syndrome. A 15-year-old female, complaining of voiding dysfunction, recurrent urinary tract infection and worsening renal function for three months. A previous history of Guillain-Barre syndrome on childhood was related. A voiding cystourethrography showed a pine-cone bladder with moderate post-void residual urine. The urodynamic demonstrated a hypocompliant bladder and small bladder capacity (190 mL) with high detrusor pressure (54 cmH2O). Nonsurgical treatments were attempted, however unsuccessfully.

  2. Pure Parsimony Xor Haplotyping

    CERN Document Server

    Bonizzoni, Paola; Dondi, Riccardo; Pirola, Yuri; Rizzi, Romeo

    2010-01-01

    The haplotype resolution from xor-genotype data has been recently formulated as a new model for genetic studies. The xor-genotype data is a cheaply obtainable type of data distinguishing heterozygous from homozygous sites without identifying the homozygous alleles. In this paper we propose a formulation based on a well-known model used in haplotype inference: pure parsimony. We exhibit exact solutions of the problem by providing polynomial time algorithms for some restricted cases and a fixed-parameter algorithm for the general case. These results are based on some interesting combinatorial properties of a graph representation of the solutions. Furthermore, we show that the problem has a polynomial time k-approximation, where k is the maximum number of xor-genotypes containing a given SNP. Finally, we propose a heuristic and produce an experimental analysis showing that it scales to real-world large instances taken from the HapMap project.

  3. Pure parsimony xor haplotyping.

    Science.gov (United States)

    Bonizzoni, Paola; Della Vedova, Gianluca; Dondi, Riccardo; Pirola, Yuri; Rizzi, Romeo

    2010-01-01

    The haplotype resolution from xor-genotype data has been recently formulated as a new model for genetic studies. The xor-genotype data is a cheaply obtainable type of data distinguishing heterozygous from homozygous sites without identifying the homozygous alleles. In this paper, we propose a formulation based on a well-known model used in haplotype inference: pure parsimony. We exhibit exact solutions of the problem by providing polynomial time algorithms for some restricted cases and a fixed-parameter algorithm for the general case. These results are based on some interesting combinatorial properties of a graph representation of the solutions. Furthermore, we show that the problem has a polynomial time k-approximation, where k is the maximum number of xor-genotypes containing a given single nucleotide polymorphisms (SNP). Finally, we propose a heuristic and produce an experimental analysis showing that it scales to real-world large instances taken from the HapMap project.

  4. Ocular Albinism Type 1 Regulates Melanogenesis in Mouse Melanocytes

    Directory of Open Access Journals (Sweden)

    Tianzhi Chen

    2016-09-01

    Full Text Available To investigate whether ocular albinism type 1 (OA1 is differentially expressed in the skin of mice with different coat colors and to determine its correlation with coat color establishment in mouse, the expression patterns and tissue distribution characterization of OA1 in the skin of mice with different coat colors were qualitatively and quantitatively analyzed by real-time quantitative PCR (qRT-PCR, immunofluorescence staining and Western blot. The qRT-PCR analysis revealed that OA1 mRNA was expressed in all mice skin samples tested, with the highest expression level in brown skin, a moderate expression level in black skin and the lowest expression level in gray skin. Positive OA1 protein bands were also detected in all skin samples by Western blot analysis. The relative expression levels of OA1 protein in both black and brown skin were significantly higher than that in gray skin, but there was no significant difference between black and brown mice. Immunofluorescence assays revealed that OA1 was mainly expressed in the hair follicle matrix, the inner and outer root sheath in the skin tissues with different coat colors. To get further insight into the important role of OA1 in the melanocytes’ pigmentation, we transfected the OA1 into mouse melanocytes and then detected the relative expression levels of pigmentation-related gene. Simultaneously, we tested the melanin content of melanocytes. As a result, the overexpression of OA1 significantly increased the expression levels of microphthalmia-associated transcription factor (MITF, tyrosinase (TYR, tyrosinase-related protein 1 (TRP1 and premelanosome protein (PMEL. However, the tyrosinase-related protein 2 (TRP2 level was attenuated. By contrast, the level of glycoprotein non-metastatic melanoma protein b (GPNMB was unaffected by OA1 overexpression. Furthermore, we observed a significant increase in melanin content in mouse melanocyte transfected OA1. Therefore, we propose that OA1 may

  5. Organizational Culture

    Directory of Open Access Journals (Sweden)

    Adrian HUDREA

    2006-02-01

    Full Text Available Cultural orientations of an organization can be its greatest strength, providing the basis for problem solving, cooperation, and communication. Culture, however, can also inhibit needed changes. Cultural changes typically happen slowly – but without cultural change, many other organizational changes are doomed to fail. The dominant culture of an organization is a major contributor to its success. But, of course, no organizational culture is purely one type or another. And the existence of secondary cultures can provide the basis for change. Therefore, organizations need to understand the cultural environments and values.

  6. Pure Laparoscopic Augmentation Ileocystoplasty

    Directory of Open Access Journals (Sweden)

    Rafael B. Rebouças

    2014-12-01

    Full Text Available Introduction Guillain-Barre syndrome is an acute neuropathy that rarely compromises bladder function. Conservative management including clean intermittent catheterization and pharmacotherapy is the primary approach for hypocompliant contracted bladder. Surgical treatment may be used in refractory cases to improve bladder compliance and capacity in order to protect the upper urinary tract. We describe a case of pure laparoscopic augmentation ileocystoplasty in a patient affected by Guillain-Barre syndrome. Presentation A 15-year-old female, complaining of voiding dysfunction, recurrent urinary tract infection and worsening renal function for three months. A previous history of Guillain-Barre syndrome on childhood was related. A voiding cystourethrography showed a pine-cone bladder with moderate post-void residual urine. The urodynamic demonstrated a hypocompliant bladder and small bladder capacity (190mL with high detrusor pressure (54 cmH2O. Nonsurgical treatments were attempted, however unsuccessfully. The patient was placed in the exaggerated Trendelenburg position. A four-port transperitoneal technique was used. A segment of ileum approximately 15-20cm was selected and divided with its pedicle. The ileal anastomosis and creation of ileal U-shaped plate were performed laparoscopically, without staplers. Bladder mobilization and longidutinal cystotomy were performed. Enterovesical anastomosis was done with continuous running suture. A suprapubic cystostomy was placed through a 5mm trocar. Results The total operative time was 335 min. The blood loss was minimal. The patient developed ileus in the early days, diet acceptance after the fourth day and was discharged on the seventh postoperative day. The urethral catheter was removed after 2 weeks. At 6-month follow-up, a cystogram showed a significant improvement in bladder capacity. The patient adhered well to clean intermittent self-catheterization and there was no report for febrile infections

  7. Coat-sleeve type giant congenital melanocytic nevus with intraoral blue nevus: A rare case report

    Directory of Open Access Journals (Sweden)

    Lata M Kale

    2017-01-01

    Full Text Available Congenital melanocytic nevi (CMN are visible hyperpigmented (melanocytic, benign, tumor like proliferations in the skin resulting from faulty development of pigment cell precursors in the embryo, and are composed of an abnormal mixture of skin elements. Giant congenital melanocytic nevus (GCMN is usually defined as a melanocytic lesion present at birth that will reach a larger size in adulthood. GCMN is a rare variety of CMN which is characterized by its size (diameter ≥20 cm and the potential for malignant transformation. It is infrequently associated with other findings, which makes the clinical picture complex. In this case, we report a rare association of GCMN with intraoral blue nevus in a 24-year-old male patient.

  8. EdnrB Governs Regenerative Response of Melanocyte Stem Cells by Crosstalk with Wnt Signaling

    Directory of Open Access Journals (Sweden)

    Makoto Takeo

    2016-05-01

    Full Text Available Delineating the crosstalk between distinct signaling pathways is key to understanding the diverse and dynamic responses of adult stem cells during tissue regeneration. Here, we demonstrate that the Edn/EdnrB signaling pathway can interact with other signaling pathways to elicit distinct stem cell functions during tissue regeneration. EdnrB signaling promotes proliferation and differentiation of melanocyte stem cells (McSCs, dramatically enhancing the regeneration of hair and epidermal melanocytes. This effect is dependent upon active Wnt signaling that is initiated by Wnt ligand secretion from the hair follicle epithelial niche. Further, this Wnt-dependent EdnrB signaling can rescue the defects in melanocyte regeneration caused by Mc1R loss. This suggests that targeting Edn/EdnrB signaling in McSCs can be a therapeutic approach to promote photoprotective-melanocyte regeneration, which may be useful for those with increased risk of skin cancers due to Mc1R variants.

  9. EdnrB governs regenerative response of melanocyte stem cells by crosstalk with Wnt signaling

    Science.gov (United States)

    Takeo, Makoto; Lee, Wendy; Rabbani, Piul; Sun, Qi; Hu, Hai; Lim, Chae Ho; Manga, Prashiela; Ito, Mayumi

    2017-01-01

    Delineating the crosstalk between distinct signaling pathways is key to understanding the diverse and dynamic responses of adult stem cells during tissue regeneration. Here we demonstrate that the Edn/EdnrB signaling pathway can interact with other signaling pathways to elicit distinct stem cell functions during tissue regeneration. EdnrB signaling promotes proliferation and differentiation of melanocyte stem cells (McSCs), dramatically enhancing the regeneration of hair and epidermal melanocytes. This effect is dependent upon active Wnt signaling that is initiated by Wnt ligand secretion from the hair follicle epithelial niche. Further, this Wnt-dependent EdnrB signaling can rescue the defects in melanocyte regeneration caused by Mc1R loss. This suggests that targeting Edn/EdnrB signaling in McSCs can be a therapeutic approach to promote photoprotective-melanocyte regeneration, which may be useful for those with increased risk of skin cancers due to Mc1R variants. PMID:27134165

  10. ARP101 inhibits α-MSH-stimulated melanogenesis by regulation of autophagy in melanocytes.

    Science.gov (United States)

    Kim, Eun Sung; Jo, Yoon Kyung; Park, So Jung; Chang, Huikyoung; Shin, Ji Hyun; Choi, Eun Sun; Kim, Jun Bum; Seok, Su Hyeon; Kim, Jae-Sung; Oh, Jeong Su; Kim, Myoung-Hwan; Lee, Eunjoo H; Cho, Dong-Hyung

    2013-12-11

    Autophagy is a cooperative process between autophagosomes and lysosomes that degrades cellular organelles. Although autophagy regulates the turnover of cellular components, its role in melanogenesis is not clearly established. Previously, we reported that ARP101 induces autophagy in various cancer cells. Here, we show that ARP101 inhibits melanogenesis by regulation of autophagy. ARP101 inhibited α-MSH-stimulated melanin synthesis and suppressed the expression of tyrosinase and TRP1 in immortalized mouse melanocytes. ARP101 also induced autophagy in melanocytes. Knockdown of ATG5 reduced both anti-melanogenic activity and autophagy mediated by ARP101 in α-MSH treated melanocytes. Electron microscopy analysis further revealed that autophagosomes engulf melanin or melanosome in α-MSH and ARP101-treated cells. Collectively, our results suggest that ARP101 inhibits α-MSH-stimulated melanogenesis through the activation of autophagy in melanocytes.

  11. NF1 loss induces senescence during human melanocyte differentiation in an iPSC-based model.

    Science.gov (United States)

    Larribere, Lionel; Wu, Huizi; Novak, Daniel; Galach, Marta; Bernhardt, Mathias; Orouji, Elias; Weina, Kasia; Knappe, Nathalie; Sachpekidis, Christos; Umansky, Ludmila; Beckhove, Philipp; Umansky, Viktor; De Schepper, Sofie; Kaufmann, Dieter; Ballotti, Robert; Bertolotto, Corine; Utikal, Jochen

    2015-07-01

    Neurofibromatosis type 1 (NF1) is a frequent genetic disease leading to the development of Schwann cell-derived neurofibromas or melanocytic lesions called café-au-lait macules (CALMs). The molecular mechanisms involved in CALMs formation remain largely unknown. In this report, we show for the first time pathophysiological mechanisms of abnormal melanocyte differentiation in a human NF1(+/-) -induced pluripotent stem cell (iPSC)-based model. We demonstrate that NF1 patient-derived fibroblasts can be successfully reprogrammed in NF1(+/-) iPSCs with active RAS signaling and that NF1 loss induces senescence during melanocyte differentiation as well as in patient's-derived CALMs, revealing a new role for NF1 in the melanocyte lineage.

  12. EdnrB Governs Regenerative Response of Melanocyte Stem Cells by Crosstalk with Wnt Signaling.

    Science.gov (United States)

    Takeo, Makoto; Lee, Wendy; Rabbani, Piul; Sun, Qi; Hu, Hai; Lim, Chae Ho; Manga, Prashiela; Ito, Mayumi

    2016-05-10

    Delineating the crosstalk between distinct signaling pathways is key to understanding the diverse and dynamic responses of adult stem cells during tissue regeneration. Here, we demonstrate that the Edn/EdnrB signaling pathway can interact with other signaling pathways to elicit distinct stem cell functions during tissue regeneration. EdnrB signaling promotes proliferation and differentiation of melanocyte stem cells (McSCs), dramatically enhancing the regeneration of hair and epidermal melanocytes. This effect is dependent upon active Wnt signaling that is initiated by Wnt ligand secretion from the hair follicle epithelial niche. Further, this Wnt-dependent EdnrB signaling can rescue the defects in melanocyte regeneration caused by Mc1R loss. This suggests that targeting Edn/EdnrB signaling in McSCs can be a therapeutic approach to promote photoprotective-melanocyte regeneration, which may be useful for those with increased risk of skin cancers due to Mc1R variants.

  13. Blood Supply--Susceptible Formation of Melanin Pigment in Hair Bulb Melanocytes of Mice

    Directory of Open Access Journals (Sweden)

    Shogo Maeda, MD

    2015-03-01

    Conclusions: Melanin pigment formation in the hair bulb melanocytes appeared to be susceptible to the blood supply, and melanocytosis was promoted in the follicles and in the epidermis of Kitl-Tg C57BL/6 mice.

  14. NFIB is a governor of epithelial-melanocyte stem cell behaviour in a shared niche.

    Science.gov (United States)

    Chang, Chiung-Ying; Pasolli, H Amalia; Giannopoulou, Eugenia G; Guasch, Géraldine; Gronostajski, Richard M; Elemento, Olivier; Fuchs, Elaine

    2013-03-07

    Adult stem cells reside in specialized niches where they receive environmental cues to maintain tissue homeostasis. In mammals, the stem cell niche within hair follicles is home to epithelial hair follicle stem cells and melanocyte stem cells, which sustain cyclical bouts of hair regeneration and pigmentation. To generate pigmented hairs, synchrony is achieved such that upon initiation of a new hair cycle, stem cells of each type activate lineage commitment. Dissecting the inter-stem-cell crosstalk governing this intricate coordination has been difficult, because mutations affecting one lineage often affect the other. Here we identify transcription factor NFIB as an unanticipated coordinator of stem cell behaviour. Hair follicle stem-cell-specific conditional targeting of Nfib in mice uncouples stem cell synchrony. Remarkably, this happens not by perturbing hair cycle and follicle architecture, but rather by promoting melanocyte stem cell proliferation and differentiation. The early production of melanin is restricted to melanocyte stem cells at the niche base. Melanocyte stem cells more distant from the dermal papilla are unscathed, thereby preventing hair greying typical of melanocyte stem cell differentiation mutants. Furthermore, we pinpoint KIT-ligand as a dermal papilla signal promoting melanocyte stem cell differentiation. Additionally, through chromatin-immunoprecipitation with high-throughput-sequencing and transcriptional profiling, we identify endothelin 2 (Edn2) as an NFIB target aberrantly activated in NFIB-deficient hair follicle stem cells. Ectopically induced Edn2 recapitulates NFIB-deficient phenotypes in wild-type mice. Conversely, endothelin receptor antagonists and/or KIT blocking antibodies prevent precocious melanocyte stem cell differentiation in the NFIB-deficient niche. Our findings reveal how melanocyte and hair follicle stem cell behaviours maintain reliance upon cooperative factors within the niche, and how this can be uncoupled in

  15. Image enhancement of optical images for binary system of melanocytes and keratinocytes

    Science.gov (United States)

    Takanezawa, S.; Baba, A.; Sako, Y.; Ozaki, Y.; Date, A.; Toyama, K.; Morita, S.

    2013-05-01

    Automatic determination of the cell shapes of large numbers of melanocytes based on optical images of human skin models have been largely unsuccessful (the complexities introduced by dendrites and the melanin pigmentation over the keratinocytes to give unclear outlines). Here, we present an image enhancement procedure for enhancing the contrast of images with removing the non-uniformity of background. The brightness is normalized also for the non-uniform population density of melanocytes.

  16. Cooperative response of keratinocytes and melanocytes to UV radiation during PUVA therapy

    Science.gov (United States)

    Stolnitz, Mikhail M.; Baskakov, Pavel V.; Peshkova, Anna Y.

    1999-03-01

    The mathematical model of processes in UV-irradiated furocoumarin-sensitized epidermis is presented taking into account the mutual influence of keratinocytes and melanocytes populations. The model describes epidermis as a hierarchical structure on tissue (keratinocytes-melanocytes cooperation, melanin screen formation), cellular (proliferation and differentiation, transitions between subpopulations), subcellular (cell movement on mitotic cycle, generation, maturing and migration of melanosomes), and molecular (melanin synthesis, processes of DNA damage and repair, molecular signal transduction) levels.

  17. Testing of viable human skin cell dilution cultures as an approach to validating microsampling.

    Science.gov (United States)

    Ainger, Stephen A; Yong, X L Hilary; Soyer, H Peter; Sturm, Richard A

    2017-05-01

    Skin biopsies are a valuable technique in the diagnosis of cutaneous inflammatory and neoplastic conditions. We were interested to test the minimal size or equivalent volume by dilution of proteolytically disassociated skin tissue required to allow the isolation and propagation of cutaneous cells, for freezing, storage and biochemical analysis. It was possible to propagate with 100% efficiency fibroblast and melanocytic cells from a 0.1 to 0.5 mm(3) equivalent neonatal foreskin sample using a combination of DispaseII and CollagenaseIV. The smallest tissue dilution that allowed melanocytic cell culture was 0.01 mm(3), which equated to approximately 16 cells based on the average skin density of melanocytes. However, passaging of cells to create frozen stocks was achieved routinely only from 1 mm(3) skin, equating to 1560 cells. Tissue-specific antigen expression of these cultures was tested by western blot of total protein extracts. There was no pigmentation antigen expression in fibroblast cultures; however, smooth muscle actin protein expression was high in fibroblast but absent from melanocytic cell strains. Melanocytic cells expressed pigmentation antigens and E-cadherin, but these were not detected in fibroblasts. Moreover, maturation of these melanocytic cells resulted in a decrease of Dopachrome Tautomerase antigen expression and induction of Tyrosinase protein consistent with the differentiation potential seen in cell cultures derived routinely from large sections of skin tissue.

  18. Melanocytic Hyperplasia in the Epidermis Overlying Trichoblastomas in 100 Randomly Selected Cases.

    Science.gov (United States)

    Al Omoush, Tahseen M M; Michal, Michael; Konstantinova, Anastasia M; Michal, Michal; Kutzner, Heinz; Kazakov, Dmitry V

    2016-04-01

    One hundred cases of trichoblastomas (large nodular, small nodular, cribriform, lymphadenoma, and columnar) were randomly selected and studied for the presence of melanocytic hyperplasia in the epidermis overlying the tumors, which was defined as foci of increased melanocytes in the basal layer of the epidermis (more than 1 per 4 basal keratinocytes). Focal melanocytic hyperplasia was detected in a total of 22 cases of trichoblastoma (22%), and this phenomenon was most frequently seen in columnar trichoblastoma (7 cases), followed by large nodular trichoblastoma (5 cases). The mechanism of epidermal melanocytic hyperplasia overlying trichoblastoma is unclear. Ultraviolet may be a contributing factor, as focal melanocytic hyperplasia was also detected in one-third of cases in the epidermis overlying uninvolved skin, usually associated with solar elastosis. This is further corroborated by the occurrence of the lesions predominantly on the face. Melanocytic hyperplasia overlying trichoblastoma appears to have no impact on the clinical appearance of the lesion and is recognized only microscopically. In an adequate biopsy specimen containing at least part of trichoblastoma, it should not cause any diagnostic problems.

  19. Understanding Melanocyte Stem Cells for Disease Modeling and Regenerative Medicine Applications.

    Science.gov (United States)

    Mull, Amber N; Zolekar, Ashwini; Wang, Yu-Chieh

    2015-12-21

    Melanocytes in the skin play an indispensable role in the pigmentation of skin and its appendages. It is well known that the embryonic origin of melanocytes is neural crest cells. In adult skin, functional melanocytes are continuously repopulated by the differentiation of melanocyte stem cells (McSCs) residing in the epidermis of the skin. Many preceding studies have led to significant discoveries regarding the cellular and molecular characteristics of this unique stem cell population. The alteration of McSCs has been also implicated in several skin abnormalities and disease conditions. To date, our knowledge of McSCs largely comes from studying the stem cell niche of mouse hair follicles. Suggested by several anatomical differences between mouse and human skin, there could be distinct features associated with mouse and human McSCs as well as their niches in the skin. Recent advances in human pluripotent stem cell (hPSC) research have provided us with useful tools to potentially acquire a substantial amount of human McSCs and functional melanocytes for research and regenerative medicine applications. This review highlights recent studies and progress involved in understanding the development of cutaneous melanocytes and the regulation of McSCs.

  20. Understanding Melanocyte Stem Cells for Disease Modeling and Regenerative Medicine Applications

    Directory of Open Access Journals (Sweden)

    Amber N. Mull

    2015-12-01

    Full Text Available Melanocytes in the skin play an indispensable role in the pigmentation of skin and its appendages. It is well known that the embryonic origin of melanocytes is neural crest cells. In adult skin, functional melanocytes are continuously repopulated by the differentiation of melanocyte stem cells (McSCs residing in the epidermis of the skin. Many preceding studies have led to significant discoveries regarding the cellular and molecular characteristics of this unique stem cell population. The alteration of McSCs has been also implicated in several skin abnormalities and disease conditions. To date, our knowledge of McSCs largely comes from studying the stem cell niche of mouse hair follicles. Suggested by several anatomical differences between mouse and human skin, there could be distinct features associated with mouse and human McSCs as well as their niches in the skin. Recent advances in human pluripotent stem cell (hPSC research have provided us with useful tools to potentially acquire a substantial amount of human McSCs and functional melanocytes for research and regenerative medicine applications. This review highlights recent studies and progress involved in understanding the development of cutaneous melanocytes and the regulation of McSCs.

  1. Immunohistochemistry and in situ hybridization in the study of human skin melanocytes.

    Science.gov (United States)

    Passeron, Thierry; Coelho, Sergio G; Miyamura, Yoshinori; Takahashi, Kaoruko; Hearing, Vincent J

    2007-03-01

    Although keratinocytes are the most numerous type of cell in the skin, melanocytes are also key players as they produce and distribute melanin that protects the skin from ultraviolet (UV) radiation. In vitro experiments on melanocytic cell lines are useful to study melanogenesis and their progression towards melanoma. However, interactions of melanocytes with keratinocytes and with other types of cells in the skin, such as fibroblasts and Langerhans cells, are also crucial. We describe two techniques, immunohistochemistry (IHC) and tissue in situ hybridization (TISH), that can be used to identify and study melanocytes in the skin and their responses to UV or other stimuli in situ. We describe a practical method to localize melanocytic antigens on formalin-fixed, paraffin-embedded tissue sections and in frozen sections using indirect immunofluorescence with conjugated secondary antibodies. In addition, we detail the use of TISH and its combination with IHC to study mRNA levels of genes expressed in the skin at cellular resolution. This methodology, along with relevant tips and troubleshooting items, are important tools to identify and study melanocytes in the skin.

  2. Melanocyte biology and function with reference to oral melanin hyperpigmentation in HIV-seropositive subjects.

    Science.gov (United States)

    Feller, Liviu; Chandran, Rakesh; Kramer, Beverley; Khammissa, Razia A G; Altini, Mario; Lemmer, Johan

    2014-09-01

    The color of normal skin and of oral mucosa is not determined by the number of melanocytes in the epithelium but rather by their melanogenic activity. Pigmented biopolymers or melanins are synthesized in melanosomes. Tyrosinase is the critical enzyme in the biosynthesis of both brown/black eumelanin and yellow/red pheomelanin. The number of the melanosomes within the melanocytes, the type of melanin within the melanosomes, and the efficacy of the transfer of melanosomes from the melanocytes to the neighboring keratinocytes all play an important role in tissue pigmentation. Melanin production is regulated by locally produced factors including proopiomelanocortin and its derivative peptides, particularly alpha-melanocyte-stimulating hormone (α-MSH), melanocortin 1 receptor (MC1R), adrenergic and cholinergic agents, growth factors, cytokines, and nitric oxide. Both eumelanin and pheomelanin can be produced by the same melanocytes, and the proportion of the two melanin types is influenced by the degree of functional activity of the α-MSH/MC1R intracellular pathway. The cause of HIV oral melanosis is not fully understood but may be associated with HIV-induced cytokine dysregulation, with the medications commonly prescribed to HIV-seropositive persons, and with adrenocortical dysfunction, which is not uncommon in HIV-seropositive subjects with AIDS. The purpose of this article is to discuss some aspects of melanocyte biology and HIV-associated oral melanin hyperpigmentation.

  3. Effect of norfloxacin and moxifloxacin on melanin synthesis and antioxidant enzymes activity in normal human melanocytes.

    Science.gov (United States)

    Beberok, Artur; Wrześniok, Dorota; Otręba, Michał; Miliński, Maciej; Rok, Jakub; Buszman, Ewa

    2015-03-01

    Fluoroquinolone antibiotics provide broad-spectrum coverage for a number of infectious diseases, including respiratory as well as urinary tract infections. One of the important adverse effects of these drugs is phototoxicity which introduces a serious limitation to their use. To gain insight the molecular mechanisms underlying the fluoroquinolones-induced phototoxic side effects, the impact of two fluoroquinolone derivatives with different phototoxic potential, norfloxacin and moxifloxacin, on melanogenesis and antioxidant enzymes activity in normal human melanocytes HEMa-LP was determined. Both drugs induced concentration-dependent loss in melanocytes viability. The value of EC50 for these drugs was found to be 0.5 mM. Norfloxacin and moxifloxacin suppressed melanin biosynthesis; antibiotics were shown to inhibit cellular tyrosinase activity and to reduce melanin content in melanocytes. When comparing the both analyzed fluoroquinolones, it was observed that norfloxacin possesses greater inhibitory effect on tyrosinase activity in melanocytes than moxifloxacin. The extent of oxidative stress in cells was assessed by measuring the activity of antioxidant enzymes: SOD, CAT, and GPx. It was observed that norfloxacin caused higher depletion of antioxidant status in melanocytes when compared with moxifloxacin. The obtained results give a new insight into the mechanisms of fluoroquinolones toxicity directed to pigmented tissues. Moreover, the presented differences in modulation of biochemical processes in melanocytes may be an explanation for various phototoxic activities of the analyzed fluoroquinolone derivatives in vivo.

  4. NMDA-R inhibition affects cellular process formation in Tilapia melanocytes; a model for pigmented adrenergic neurons in process formation and retraction.

    Science.gov (United States)

    Ogundele, Olalekan Michael; Okunnuga, Adetokunbo Adedotun; Fabiyi, Temitope Deborah; Olajide, Olayemi Joseph; Akinrinade, Ibukun Dorcas; Adeniyi, Philip Adeyemi; Ojo, Abiodun Ayodele

    2014-06-01

    Parkinson's disease has long been described to be a product of dopamine and (or) melanin loss in the substanstia nigra (SN). Although most studies have focused on dopaminergic neurons, it is important to consider the role of pigment cells in the etiology of the disease and to create an in vitro live cell model for studies involving pigmented adrenergic cells of the SN in Parkinsonism. The Melanocytes share specific features with the pigmented adrenergic neurons as both cells are pigmented, contain adrenergic receptors and have cellular processes. Although the melanocyte cellular processes are relatively short and observable only when stimulated appropriately by epinephrine and other factors or molecules. This study employs the manipulation of N-Methyl-D-Aspartate Receptor (NMDA-R), a major receptor in neuronal development, in the process formation pattern of the melanocyte in order to create a suitable model to depict cellular process elongation and shortening in pigmented adrenergic cells. NMDA-R is an important glutamate receptor implicated in neurogenesis, neuronal migration, maturation and cell death, thus we investigated the role of NMDA-R potentiation by glutamate/KCN and its inhibition by ketamine in the behavior of fish scale melanocytes in vitro. This is aimed at establishing the regulatory role of NMDA-R in this cell type (melanocytes isolated form Tilapia) in a similar manner to what is observable in the mammalian neurons. In vitro live cell culture was prepared in modified Ringer's solution following which the cells were treated as follows; Control, Glutamate, Ketamine, Glutamate + Ketamine, KCN + Ketamine and KCN. The culture was maintained for 10 min and the changes were captured in 3D-Time frame at 0, 5 and 10 min for the control and 5, 7 and 10 min for each of the treatment category. Glutamate treatment caused formation of short cellular processes localized directly on the cell body while ketamine treatment (inhibition of NMDA-R) facilitated

  5. Effects of Lutein and Zeaxanthin on LPS-Induced Secretion of IL-8 by Uveal Melanocytes and Relevant Signal Pathways

    Directory of Open Access Journals (Sweden)

    Shih-Chun Chao

    2015-01-01

    Full Text Available The effects of lutein and zeaxanthin on lipopolysaccharide- (LPS- induced secretion of IL-8 by uveal melanocytes (UM were tested in cultured human UM. MTT assay revealed that LPS (0.01–1 μg/mL and lutein and zeaxanthin (1–10 μM did not influence the cell viability of cultured UM. LPS caused a dose-dependent increase of secretion of IL-8 by cultured UM. Lutein and zeaxanthin did not affect the constitutive secretion of IL-8. However, lutein and zeaxanthin decreased LPS-induced secretion of IL-8 in cultured UM in a dose-dependent manner. LPS significantly increased NF-κB levels in cell nuclear extracts and p-JNK levels in the cell lysates from UM, but not p-p38 MAPK and p-ERG. Lutein or zeaxanthin significantly reduced LPS-induced increase of NF-κB and p-JNK levels, but not p38 MAPK and ERG levels. The present study demonstrated that lutein and zeaxanthin inhibited LPS-induced secretion of IL-8 in cultured UM via JNK and NF-κB signal pathways. The anti-inflammatory effects of lutein and zeaxanthin might be explored as a therapeutic approach in the management of uveitis and other inflammatory diseases of the eye.

  6. Effects of Lutein and Zeaxanthin on LPS-Induced Secretion of IL-8 by Uveal Melanocytes and Relevant Signal Pathways.

    Science.gov (United States)

    Chao, Shih-Chun; Vagaggini, Tommaso; Nien, Chan-Wei; Huang, Sheng-Chieh; Lin, Hung-Yu

    2015-01-01

    The effects of lutein and zeaxanthin on lipopolysaccharide- (LPS-) induced secretion of IL-8 by uveal melanocytes (UM) were tested in cultured human UM. MTT assay revealed that LPS (0.01-1 μg/mL) and lutein and zeaxanthin (1-10 μM) did not influence the cell viability of cultured UM. LPS caused a dose-dependent increase of secretion of IL-8 by cultured UM. Lutein and zeaxanthin did not affect the constitutive secretion of IL-8. However, lutein and zeaxanthin decreased LPS-induced secretion of IL-8 in cultured UM in a dose-dependent manner. LPS significantly increased NF-κB levels in cell nuclear extracts and p-JNK levels in the cell lysates from UM, but not p-p38 MAPK and p-ERG. Lutein or zeaxanthin significantly reduced LPS-induced increase of NF-κB and p-JNK levels, but not p38 MAPK and ERG levels. The present study demonstrated that lutein and zeaxanthin inhibited LPS-induced secretion of IL-8 in cultured UM via JNK and NF-κB signal pathways. The anti-inflammatory effects of lutein and zeaxanthin might be explored as a therapeutic approach in the management of uveitis and other inflammatory diseases of the eye.

  7. CDK5对羊驼皮肤黑色素细胞TYR和MITF mRNA表达的调节%Regulation of TYR and MITF mRNA Expression by CDK5 in Alpaca Melanocytes

    Institute of Scientific and Technical Information of China (English)

    张瑞娜; 范瑞文; 程志学; 田雪; 刘佳; 高磊; 马峥; 董常生

    2011-01-01

    为了证实CDK5是否参与羊驼毛色的形成,本研究主要对CDK5在羊驼黑色素细胞中调节TYR和MI TF的表达进行了研究.本研究首先采用免疫组织化学方法检测CDK5在羊驼皮肤黑色素细胞中的定位,再通过脂质体将CDK5转染于羊驼皮肤黑色素细胞,之后通过Western blot和qRT-PCR的方法检测转染后黑色素细胞中CDK5蛋白、TYR和MITF mRNA的表达.免疫组化结果显示CDK5位于黑色素细胞的胞质和细胞核内;Western blot结果显示转染组黑色素细胞中CDK5蛋白表达量明显高于对照组;qRT-PCR结果显示CDK5可下调MITF的表达,同时上调TYR的表达,转染组黑色素细胞中MITF和TYR mRNA的表达水平分别是对照组细胞的0.264 9和3.931 3倍.结果揭示CDK5可能通过调节黑色素细胞核中TYR和MITF mRNA的表达,从而参与调控羊驼毛色形成.%In order to investigate whether CDK5 could participate in the coat color formation of alpaca, the regulation of CDK5 on TYR and MITF expression in alpaca melanocytes was tested in this study. The localization of CDK5 in cultured alpaca melanocytes was examined by immunohis-tochemistry. Then CDK5 was transfected into cultured alpaca melanocytes by liposomes. Following the transfection, CDK5 protein was detected by Western blot and the expression of MITF and TYR was detected by quantitative real-time PCR(qRT-PCR). The immunohistochemistry results showed that CDK5 was expressed in cytoplasm and nucleus of cultured alpaca melanocytes; Western blot results showed that the level of CDK5 protein was apparently higher in transfected melanocytes than that in normal melanocytes, and qRT-PCR results showed that the mRNA abundance of MITF was down-regulated while the mRNA abundance of TYR was up-regulated by the transfection with CDK5, and the expression level of MITF and TYR mRNA in transfected melanocytes was 0. 264 9 and 3. 931 3 folds higher than that in controlled melanocytes respectively. The results suggest

  8. Visual processing in pure alexia

    DEFF Research Database (Denmark)

    Starrfelt, Randi; Habekost, Thomas; Gerlach, Christian

    2010-01-01

    Whether pure alexia is a selective disorder that affects reading only, or if it reflects a more general visual disturbance, is highly debated. We have investigated the selectivity of visual deficits in a pure alexic patient (NN) using a combination of psychophysical measures, mathematical modelling...

  9. The Effects of Endothelin-3 on the Melanocyte Characteristic and Expression of Hair Color Genes in Melanocytes from Alpaca (Lama Pacos) in vitro%内皮素-3对羊驼黑色素细胞特征及细胞内毛色基因表达的影响

    Institute of Scientific and Technical Information of China (English)

    耿建军; 白俊明; 范瑞文; 杨姗姗; 庞亚妙; 董常生

    2013-01-01

    The study aimed to investigate the effects of endothelin-3 (EDN3) on the expression of hair color genes of alpaca.The activity of melanocyte,the production of melanin in melanocytes and the expressions of related genes and proteins including Endothelin receptor B (EDNRB),KIT,Microphthalmia-associtated transcription factor (MITF) and Tyrosinase (TYR) in alpaca melanocytes in cultures with addition of different dose of EDN3(0,10-9,10-8,10-7mol · L-1),were examined by MTT,ultraviolet spectrophotometry,qRT-PCR and Western blotting,respectively.The results showed that the amount of melanocytes was increased with the increasing double-or tri-dendrites; At the addition dose of 10-8 mol · L-1 for 72 h after adding EDN3,compared to control group,the proliferation of melanocytes was obvious and the expressions of EDNRB,KIT,MITF and TYR significantly up-regulated at the level of both transcription and translation in melanocytes(P<0.05).Moreover,the production of melanin in melanocytes significantly increased(P<0.05).These findings indicate that EDN3 play an important role in regulating melanins production in melanocytes in alpaca.%为了研究内皮素-3(Endothelin-3,EDN3)对羊驼皮肤黑色素细胞内毛色形成相关基因的影响.本研究在体外培养的羊驼皮肤黑色素细胞中添加不同浓度(0、10-9、10-8、10-7mol· L-1)的EDN3,通过MTT法、紫外分光光度法、qRT-PCR和Western blotting技术分别检测黑色素细胞活力、黑色素产量、相关基因和蛋白(包括内皮素受体B(Endothelin receptor B,EDNRB)、KIT、小眼畸形相关转录因子(Microphthalmia-associtated transcription factor,MITF)和酪氨酸酶(Tyrosinase,TYR))的表达情况.结果表明,在羊驼皮肤黑色素细胞添加EDN3 72 h 后,黑色素细胞呈双树突或三树突状,且细胞数量明显增加;在添加适当浓度10-8mol·L-1时,细胞具有明显的增殖,细胞内EDNRB、KIT、MITF和TRY在转录水平和蛋白水平的表达量被上调,

  10. 黑素细胞痣的形成机制%Formation mechanism of melanocytic nevi

    Institute of Scientific and Technical Information of China (English)

    严军华; 张美华

    2010-01-01

    Melanocytic nevi are common in clinic, while its mechanism is unknown yet. Recent studies on relation between eruptive melanocytic nevi and immunosuppressive agents have shown that the formation of melanocytic nevi is relevant with immunosuppression. Melanocyte-stimulating hormone (MSH)-α plays a certain role in the development of melanocytic nevi by up-regulating the expression of microphthalmia-associated transcription factor (Mitf) and promoting the proliferation of melanocytes through cAMP pathway. Also, MSH-α together with immunosuppressive agents could affect Langerhans cells,T cells and expressions of cytokines such as interleukin (IL)-10, IL-12, etc., possibly exert upon MAPK pathway, up-regulate the expression of Mitf, thereby result in the formation of melanocytic nevi.%黑素细胞痣临床常见,通过对发疹性黑素细胞痣与免疫抑制剂的关系分析,认为黑素细胞痣的产生与免疫抑制有关.促黑素细胞激素-α能促进黑素细胞的增殖,在黑素细胞痣形成中发挥作用.其可通过环磷腺苷途径上调小眼畸形相关转录因子的表达,促进黑素细胞的增殖.促黑素细胞激素-α和免疫抑制剂影响朗格汉斯细胞、T细胞及IL-10、IL-12等细胞因子,可能作用于促分裂原激活蛋白激酶信号通路,上调小眼畸形相关转录因子的表达,从而导致黑素细胞痣的形成.

  11. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  12. Clinical dermatoscopy of big melanocytic naevuses in patients with psoriasis

    Directory of Open Access Journals (Sweden)

    Makarova N.G.

    2012-06-01

    Full Text Available

    The study aimed to determine clinical characteristics of dermatoscopy of big melanocytic naevuses (MN in patients with psoriasis. Methods. The were 32 patients with psoriasis (11 women at an average age of 37,4 years and 100 people (79 women at an average age of 27,8 years with no psoriasis. Both groups had a survey and full photometry screening of skin. Besides Fitzpatrick skin typing test was used, pigmentary formations were examined using dermato-scope HEINE MINI 3000 10X with 70 % ethyl alcohol immersion. Naevuses were estimated by ABCD and ABC scales. Results. MN more than 5 mm in diameter are more often in patients with psoriasis (68,8 % than in control group (37 %. However in control group there are less naevuses of appropriate dermatological dysplastic formations criteria (32 % comparison to the group without psoriasis (42 %. There is an important fact that patients with psoriasis often have 2-phototype of skin and artifcial insolation (tanning bed, PUVA, narrow-band phototherapy 311 nm. Conclusion. 68,8 % patients have MN more than 5 mm in diameter. 32 % of patients with psoriasis had MN of dysplastic naevus criteria according to Pehamberger. In one patient naevus was of appropriate malignant tumors criteria according to ABCD and ABC scales.

  13. Oxidative stress-induced overexpression of miR-25: the mechanism underlying the degeneration of melanocytes in vitiligo.

    Science.gov (United States)

    Shi, Q; Zhang, W; Guo, S; Jian, Z; Li, S; Li, K; Ge, R; Dai, W; Wang, G; Gao, T; Li, C

    2016-03-01

    Oxidative stress has a critical role in the pathogenesis of vitiligo. However, the specific molecular mechanism involved in oxidative stress-induced melanocyte death is not well characterized. Given the powerful role of microRNAs (miRNAs) in the regulation of cell survival as well as the fact that the generation of miRNAs can be affected by oxidative stress, we hypothesized that miRNAs may participate in vitiligo pathogenesis by modulating the expression of vital genes in melanocytes. In the present study, we initially found that miR-25 was increased in both serum and lesion samples from vitiligo patients, and its serum level was correlated with the activity of vitiligo. Moreover, restoration of miR-25 promoted the H2O2-induced melanocyte destruction and led to the dysfunction of melanocytes. Further experiments proved that MITF, a master regulator in melanocyte survival and function, accounted for the miR-25-caused damaging impact on melanocytes. Notably, other than the direct role on melanocytes, we observed that miR-25 inhibited the production and secretion of SCF and bFGF from keratinocytes, thus impairing their paracrine protective effect on the survival of melanocytes under oxidative stress. At last, we verified that oxidative stress could induce the overexpression of miR-25 in both melanocytes and keratinocytes possibly by demethylating the promoter region of miR-25. Taken together, our study demonstrates that oxidative stress-induced overexpression of miR-25 in vitiligo has a crucial role in promoting the degeneration of melanocytes by not only suppressing MITF in melanocytes but also impairing the paracrine protective effect of keratinocytes. Therefore, it is worthy to investigate the possibility of miR-25 as a potential drug target for anti-oxidative therapy in vitiligo.

  14. The redox antimalarial dihydroartemisinin targets human metastatic melanoma cells but not primary melanocytes with induction of NOXA-dependent apoptosis.

    Science.gov (United States)

    Cabello, Christopher M; Lamore, Sarah D; Bair, Warner B; Qiao, Shuxi; Azimian, Sara; Lesson, Jessica L; Wondrak, Georg T

    2012-08-01

    Recent research suggests that altered redox control of melanoma cell survival, proliferation, and invasiveness represents a chemical vulnerability that can be targeted by pharmacological modulation of cellular oxidative stress. The endoperoxide artemisinin and semisynthetic artemisinin-derivatives including dihydroartemisinin (DHA) constitute a major class of antimalarials that kill plasmodium parasites through induction of iron-dependent oxidative stress. Here, we demonstrate that DHA may serve as a redox chemotherapeutic that selectively induces melanoma cell apoptosis without compromising viability of primary human melanocytes. Cultured human metastatic melanoma cells (A375, G361, LOX) were sensitive to DHA-induced apoptosis with upregulation of cellular oxidative stress, phosphatidylserine externalization, and activational cleavage of procaspase 3. Expression array analysis revealed DHA-induced upregulation of oxidative and genotoxic stress response genes (GADD45A, GADD153, CDKN1A, PMAIP1, HMOX1, EGR1) in A375 cells. DHA exposure caused early upregulation of the BH3-only protein NOXA, a proapototic member of the Bcl2 family encoded by PMAIP1, and genetic antagonism (siRNA targeting PMAIP1) rescued melanoma cells from apoptosis indicating a causative role of NOXA-upregulation in DHA-induced melanoma cell death. Comet analysis revealed early DHA-induction of genotoxic stress accompanied by p53 activational phosphorylation (Ser 15). In primary human epidermal melanocytes, viability was not compromised by DHA, and oxidative stress, comet tail moment, and PMAIP1 (NOXA) expression remained unaltered. Taken together, these data demonstrate that metastatic melanoma cells display a specific vulnerability to DHA-induced NOXA-dependent apoptosis and suggest feasibility of future anti-melanoma intervention using artemisinin-derived clinical redox antimalarials.

  15. Establishment of coculture system of normal human epidermal keratinocytes and melanocytes in vitro%正常人角质形成细胞和黑素细胞体外共培养体系的建立

    Institute of Scientific and Technical Information of China (English)

    李海东; 王鹰

    2012-01-01

    目的 体外建立正常人表皮角质形成细胞和黑素细胞直接接触的共培养体系.方法 分别培养角质形成细胞和黑素细胞,体外建立角质形成细胞和黑素细胞直接接触的共培养模型,采用四甲基偶氮唑盐(MTT)比色法、紫外分光光度计检测、左旋多巴(L-Dopa)染色、免疫组织化学染色及透射电镜观察对共培养体系中的黑素细胞和角质形成细胞进行生物学鉴定.结果 单独培养时,角质形成细胞呈圆形、"铺路石样"生长,黑素细胞呈两极或多级树突状生长;角质形成细胞和黑素细胞按一定比例混合培养后,2种细胞均迅速增殖,黑素细胞树突多为3~5个,与数十个角质形成细胞呈团块状生长,形成类似黑素单元的结构.共培养体系中的黑素细胞和角质形成细胞具备正常的生物学功能.结论 角质形成细胞和黑素细胞直接接触的共培养体系为黑素细胞提供了更接近生理状态的体外研究模型.%Objective To establish coculture system of direct cell-to-cell contact between normal human epidermal keratinocytes and melanocytes in vitro. Methods Keratinocytes and melanocytes were cultured respectively,and then the coculture model of direct cell-to-cell contact between keratinocytes and melanocytes was established in vilro. Methyl thiazolyl tetrazolium(MTT) assay, UV spectrophotometry,levoDopa(L-Dopa) staining, immunohistochemical staining and transmission electron microscopy observation were employed to biologically identify the melanocytes and keratinocytes in the coculture system. Results Keratinocytes were round and grew in "cobblestone" pattern while melanocytes in bipolar or multi-level dendritic growth pattern when they were cultured separately. The 2 kinds of cells proliferated rapidly when they cocultured in a certain proportion. Melanocytes with 3 to 5 den-drites grew in crumb pattern with dozens of keratinocyte,similar to the structure of melanin unit. Melanocytes

  16. Lacunarity analysis: a promising method for the automated assessment of melanocytic naevi and melanoma.

    Science.gov (United States)

    Gilmore, Stephen; Hofmann-Wellenhof, Rainer; Muir, Jim; Soyer, H Peter

    2009-10-13

    The early diagnosis of melanoma is critical to achieving reduced mortality and increased survival. Although clinical examination is currently the method of choice for melanocytic lesion assessment, there is a growing interest among clinicians regarding the potential diagnostic utility of computerised image analysis. Recognising that there exist significant shortcomings in currently available algorithms, we are motivated to investigate the utility of lacunarity, a simple statistical measure previously used in geology and other fields for the analysis of fractal and multi-scaled images, in the automated assessment of melanocytic naevi and melanoma. Digitised dermoscopic images of 111 benign melanocytic naevi, 99 dysplastic naevi and 102 melanomas were obtained over the period 2003 to 2008, and subject to lacunarity analysis. We found the lacunarity algorithm could accurately distinguish melanoma from benign melanocytic naevi or non-melanoma without introducing many of the limitations associated with other previously reported diagnostic algorithms. Lacunarity analysis suggests an ordering of irregularity in melanocytic lesions, and we suggest the clinical application of this ordering may have utility in the naked-eye dermoscopic diagnosis of early melanoma.

  17. Lacunarity analysis: a promising method for the automated assessment of melanocytic naevi and melanoma.

    Directory of Open Access Journals (Sweden)

    Stephen Gilmore

    Full Text Available The early diagnosis of melanoma is critical to achieving reduced mortality and increased survival. Although clinical examination is currently the method of choice for melanocytic lesion assessment, there is a growing interest among clinicians regarding the potential diagnostic utility of computerised image analysis. Recognising that there exist significant shortcomings in currently available algorithms, we are motivated to investigate the utility of lacunarity, a simple statistical measure previously used in geology and other fields for the analysis of fractal and multi-scaled images, in the automated assessment of melanocytic naevi and melanoma. Digitised dermoscopic images of 111 benign melanocytic naevi, 99 dysplastic naevi and 102 melanomas were obtained over the period 2003 to 2008, and subject to lacunarity analysis. We found the lacunarity algorithm could accurately distinguish melanoma from benign melanocytic naevi or non-melanoma without introducing many of the limitations associated with other previously reported diagnostic algorithms. Lacunarity analysis suggests an ordering of irregularity in melanocytic lesions, and we suggest the clinical application of this ordering may have utility in the naked-eye dermoscopic diagnosis of early melanoma.

  18. Melanocytes and keratinocytes have distinct and shared responses to ultraviolet radiation and arsenic.

    Science.gov (United States)

    Cooper, K L; Yager, J W; Hudson, L G

    2014-01-30

    The rise of melanoma incidence in the United States is a growing public health concern. A limited number of epidemiology studies suggest an association between arsenic levels and melanoma risk. Arsenic acts as a co-carcinogen with ultraviolet radiation (UVR) for the development of squamous cell carcinoma and proposed mechanisms include generation of oxidative stress by arsenic and UVR and inhibition of UVR-induced DNA repair by arsenic. In this study, we investigate similarities and differences in response to arsenic and UVR in keratinocytes and melanocytes. Normal melanocytes are markedly more resistant to UVR-induced cytotoxicity than normal keratinocytes, but both cell types are equally sensitive to arsenite. Melanocytes were more resistant to arsenite and UVR stimulation of superoxide production than keratinocytes, but the concentration of arsenite necessary to inhibit the activity of the DNA repair protein poly(ADP-ribose)polymerase and enhance retention of UVR-induced DNA damage was essentially equivalent in both cell types. These findings suggest that although melanocytes are less sensitive than keratinocytes to initial UVR-mediated DNA damage, both of these important target cells in the skin share a mechanism related to arsenic inhibition of DNA repair. These findings suggest that concurrent chronic arsenic exposure could promote retention of unrepaired DNA damage in melanocytes and act as a co-carcinogen in melanoma.

  19. The Long Noncoding RNA SPRIGHTLY Regulates Cell Proliferation in Primary Human Melanocytes.

    Science.gov (United States)

    Zhao, Wei; Mazar, Joseph; Lee, Bongyong; Sawada, Junko; Li, Jian-Liang; Shelley, John; Govindarajan, Subramaniam; Towler, Dwight; Mattick, John S; Komatsu, Masanobu; Dinger, Marcel E; Perera, Ranjan J

    2016-04-01

    The long noncoding RNA SPRIGHTLY (formerly SPRY4-IT1), which lies within the intronic region of the SPRY4 gene, is up-regulated in human melanoma cells compared to melanocytes. SPRIGHTLY regulates a number of cancer hallmarks, including proliferation, motility, and apoptosis. To better understand its oncogenic role, SPRIGHTLY was stably transfected into human melanocytes, which resulted in increased cellular proliferation, colony formation, invasion, and development of a multinucleated dendritic-like phenotype. RNA sequencing and mass spectrometric analysis of SPRIGHTLY-expressing cells revealed changes in the expression of genes involved in cell proliferation, apoptosis, chromosome organization, regulation of DNA damage responses, and cell cycle. The proliferation marker Ki67, minichromosome maintenance genes 2-5, antiapoptotic gene X-linked inhibitor of apoptosis, and baculoviral IAP repeat-containing 7 were all up-regulated in SPRIGHTLY-expressing melanocytes, whereas the proapoptotic tumor suppressor gene DPPIV/CD26 was down-regulated, followed by an increase in extracellular signal-regulated kinase 1/2 phosphorylation, suggesting an increase in mitogen-activated protein kinase activity. Because down-regulation of DPPIV is known to be associated with malignant transformation in melanocytes, SPRIGHTLY-mediated DPPIV down-regulation may play an important role in melanoma pathobiology. Together, these findings provide important insights into how SPRIGHTLY regulates cell proliferation and anchorage-independent colony formation in primary human melanocytes.

  20. Differential PAX3 functions in normal skin melanocytes and melanoma cells.

    Science.gov (United States)

    Medic, Sandra; Rizos, Helen; Ziman, Mel

    2011-08-12

    The PAX3 transcription factor is the key regulator of melanocyte development during embryogenesis and is also frequently found in melanoma cells. While PAX3 is known to regulate melanocyte differentiation, survival, proliferation and migration during development, it is not clear if its function is maintained in adult melanocytes and melanoma cells. To clarify this we have assessed which genes are targeted by PAX3 in these cells. We show here that similar to its roles in development, PAX3 regulates complex differentiation networks in both melanoma cells and melanocytes, in order to maintain cells as "stem" cell-like (via NES and SOX9). We show also that mediators of migration (MCAM and CSPG4) are common to both cell types but more so in melanoma cells. By contrast, PAX3-mediated regulation of melanoma cell proliferation (through TPD52) and survival (via BCL2L1 and PTEN) differs from that in melanocytes. These results suggest that by controlling cell proliferation, survival and migration as well as maintaining a less differentiated "stem" cell like phenotype, PAX3 may contribute to melanoma development and progression.

  1. [The intervention of nicotinamide on skin melanocyte's cell proliferation after UVA (365 nm) exposed.].

    Science.gov (United States)

    Patam, Muhammad; Jin, Xi-peng; Pan, Jian-ying; Shen, Guang-zu; Jin, Tai-Yi

    2005-02-01

    To investigate the interference effect of nicotinamide on UVA-induced cell proliferation in human skin melanocyte. To apply the optimum UVA dose expected to cause cell proliferation: 0.2 cm2, nicotinamide was added after the 0.2 cm2 UVA exposure immediately or 48 h later, then the rate of cell proliferation, calcium concentration and the activities of Na+-K+, Ca2+-ATP enzymes of melanocytes were measured respectively. After treatment with 1.000 mg/ml nicotinamide following UVA exposure, the rate of cell proliferation was decreased significantly 24 hours later. Treatment with 0.125 mg/ml nicotinamide 48 hours after UVA exposure also significantly inhibited the cell proliferation; 1.25 mg/ml nicotinamide increased calcium concentration in cells; 0.250 mg/ml nicotinamide increased the activities of Na+-K+, Ca2+-ATP enzymes in melanocytes (P Nicotinamide has more obvious effect on inhibiting melanocyte's proliferation if added immediately following UVA exposure. Our discovery indicated that nicotinamide may affect the melanocyte through modulating the calcium concentration. It is possible to consider nicotinamide as an efficient and safe sun screen to provide a certain level of protection for UVA exposed skin.

  2. Treatment of a giant congenital melanocytic nevus in the adult: review of the current management of giant congenital melanocytic nevus.

    Science.gov (United States)

    Su, Jeannie J; Chang, Daniel K; Mailey, Brian; Gosman, Amanda

    2015-05-01

    Giant congenital melanocytic nevi (GCMNs) create cosmetic disfigurements and pose risk for malignant transformation. Adult GCMN cases are uncommon because most families opt for surgical treatment during childhood. We review the current literature on GCMN and present an interesting case of an adult with a GCMN encompassing the entire back with painful nodules exhibiting gross involvement of his back musculature, without pathologic evidence of malignancy. Surgical management was deferred in childhood because of parental desires to allow the patient to make his own decision, and treatment in adulthood was pursued on the basis of the significant impairment of the patient's quality of life and self-esteem due to the massive size and deforming nature of the nevus. The treatment strategy used for this young adult male patient involved a massive en bloc excision of the GCMN with partial resection of the latissimus dorsi, followed by a 5-week staged reconstructive process using dermal regenerative matrices and split-thickness skin grafting. Because of the shift in GCMN management from early surgical management to more conservative management, we may see an increase in adult cases of GCMN. Thus, it is critical to better understand the controversy surrounding early versus delayed management of GCMN.

  3. 玻璃酸钠对黑素细胞生物学活性的影响%Effects of hyaluronic acid on biological activity of melanocytes

    Institute of Scientific and Technical Information of China (English)

    洪为松; 钱国培; 许爱娥

    2011-01-01

    Objective To observe the effect of hyaluronic acid on the proliferation of and tyrosinase activity in melanocytes.Methods Normal primary human melanocytes were isolated from infant foreskin tissue and cultured.Different concentrations(0 to 10 g/L)of hyaluronic acid wero added to the culture medium immediately or 8 hours after the inoculation of melanocytes.MTT assay was performed to detect the proliferation of melanocytes,and tyrosinase activity was determined to evaluate the effect of hyaluronic acid on the melanin synthesis by melanocytes.Results The proliferation level (absorbance at 490 am,A490)of melanocytes was 0.14±0.02,0.37±0.08,0.45±0.11,0.49±0.07,0.55±0.12,0.52±0.11,0.49±0.07,0.39±0.05,0.19±0.03 and 0.01 4-0.01 when treated with hyaluronic acid of 0,0.008,0.016,0.313,0.625,1.250,2.500,5.000,7.500 and 10.000 g/L,respectively.The hyaluronic acid of 0.08 to 5 g/L markedly accelerated the proliferation of melanocytes,while that of 10 g/L inhibited their proliferation.The tyrosinase activity in melanocytes was promoted by hyaluronie acid of 0.2 to 5 g/L,but suppressed by that of 10 g/L.The proliferation of melanocytes treated with hyaluronic acid immediately after the inoculation was more rapid than that treated with hyaluronic acid 8 hours after the inoculation.Conclusion The hyaluronic acid of 0.2 to 5 g/L can enhance the proliferation of and tyrosinase activity in melanocytes.%目的 观察玻璃酸钠对黑素细胞增殖及酪氨酸酶活性的影响.方法 噻唑蓝法和酪氨酸酶活性测定法分别检测不同浓度玻璃酸钠对正常人黑素细胞生长和黑素合成能力的影响.结果 玻璃酸钠质量浓度为0,0.008、0.016、0.313、0.625、1.250、2.500、5.000、7.500、10.000 g/L时,黑素细胞增殖活性(A490)分别为0.14±0.02、0.37±0.08、0.45±0.11、0.49±0.07、0.55±0.12、0.52±0.11、0.49±0.07、0.39±0.05、0.19±0.03、0.01±0.01,玻璃酸钠质量浓度在0.08~5 g/L范围内时,对黑素细胞的

  4. Influences of infrared irradiation on human epidermal melanocytes%红外线照射对人表皮黑素细胞影响的研究

    Institute of Scientific and Technical Information of China (English)

    张悦; 赵广; 孟如松; 宋文婷; 邵丽芳; 牛建荣

    2012-01-01

    目的 探讨红外线(infrared,JR)照射促进体外培养的正常人表皮黑素细胞的酪氨酸酶活性及黑素含量的作用,寻找IR照射的最佳剂量.方法 将体外培养的正常人表皮黑素细胞分别给予不同的红外线剂量:0 J/cm2(对照组)、20 J/cm2、60 J/cm2、80 J/cm2、100 J/cm2、140 J/cm2、240 J/cm2、320 J/cm2,采用四甲基偶氮唑蓝(MTT)法选择最佳剂量并以此作为照射条件,连续3d,检测黑素细胞的酪氨酸酶活性、酪氨酸酶活性增长率、黑素含量、红外线照射对黑素细胞影响率以及细胞周期.结果 红外线照射的最佳剂量为80 J/cm2,此照射下的酪氨酸酶活性0.3601±0.0301,高于对照组0.3114±0.0341 (P< 0.05),酪氨酸酶活性增长率为15.64%.照射组的黑素细胞含量为0.2748±0.0243,高于对照组0.2325±0.0254(P<0.05),红外线照射对黑素细胞黑素含量影响率18.19%.细胞周期检测结果显示,G1细胞比例明显低于对照组(P<0.01),G2期和S期细胞比例较对照组增加(P<0.05).结论 80 J/cm2是IR照射的理想剂量,可以增加黑素细胞的黑素含量,提高酪氨酸酶活性,促进黑素细胞分化增殖.%Objective To observe the increasing effect of infrared irradiation on tyrosinase activity and melanin content in cultured normal human epidermal melanocytes in vitro and to explore the optimal dose of infrared irradiation.Methods Epidermal melanocytes were isolated from normal human foreskin tissue,and subjected to primary culture.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the effect of different doses(0,20,60,80,100,140,240,320 J/cm2)of infrared light on the proliferation of melanocytes and to select the optimal irradiation dose.Then,melanocytes were irradiated with infrared light at the optimal dose for 3 consecutive days followed by the determination of tyrosinase activity,melanin content,and cell cycle via dopa oxidation assay,NaOH solubilization method and flow cytometry

  5. Distinct phenotypic changes between the superficial and deep component of giant congenital melanocytic naevi: a rationale for curettage.

    NARCIS (Netherlands)

    Raeve, L.E. De; Claes, A.; Ruiter, D.J.; Muijen, G.N.P. van; Roseeuw, D.; Kempen, L.C. van

    2006-01-01

    BACKGROUND: Giant congenital melanocytic naevi (GCMN) convey a 14-fold increased melanoma risk. In contrast, medium congenital melanocytic naevi (MCMN) are rarely associated with malignant transformation. Management of patients with GCMN is challenging and there is no consensus on the most appropria

  6. Activating mutations of the GNAQ gene : a frequent event in primary melanocytic neoplasms of the central nervous system

    NARCIS (Netherlands)

    Kusters-Vandevelde, Heidi V. N.; Klaasen, Annelies; Kusters, Benno; Groenen, Patricia J. T. A.; van Engen-van Grunsven, Ilse A. C. H.; van Dijk, Marcory R. C. F.; Reifenberger, Guido; Wesseling, Pieter; Blokx, Willeke A. M.

    2010-01-01

    Primary melanocytic neoplasms of the central nervous system (CNS) are uncommon neoplasms derived from melanocytes that normally can be found in the leptomeninges. They cover a spectrum of malignancy grades ranging from low-grade melanocytomas to lesions of intermediate malignancy and overtly maligna

  7. The nuclear factor (erythroid-derived 2)-like 2 (NRF2) antioxidant response promotes melanocyte viability and reduces toxicity of the vitiligo-inducing phenol monobenzone.

    Science.gov (United States)

    Arowojolu, Omotayo A; Orlow, Seth J; Elbuluk, Nada; Manga, Prashiela

    2017-07-01

    Vitiligo, characterised by progressive melanocyte death, can be initiated by exposure to vitiligo-inducing phenols (VIPs). VIPs generate oxidative stress in melanocytes and activate the master antioxidant regulator NRF2. While NRF2-regulated antioxidants are reported to protect melanocytes from oxidative stress, the role of NRF2 in the melanocyte response to monobenzone, a clinically relevant VIP, has not been characterised. We hypothesised that activation of NRF2 may protect melanocytes from monobenzone-induced toxicity. We observed that knockdown of NRF2 or NRF2-regulated antioxidants NQO1 and PRDX6 reduced melanocyte viability, but not viability of keratinocytes and fibroblasts, suggesting that melanocytes were preferentially dependent upon NRF2 activity for growth compared to other cutaneous cells. Furthermore, melanocytes activated the NRF2 response following monobenzone exposure and constitutive NRF2 activation reduced monobenzone toxicity, supporting NRF2's role in the melanocyte stress response. In contrast, melanocytes from individuals with vitiligo (vitiligo melanocytes) did not activate the NRF2 response as efficiently. Dimethyl fumarate-mediated NRF2 activation protected normal and vitiligo melanocytes against monobenzone-induced toxicity. Given the contribution of oxidant-antioxidant imbalance in vitiligo, modulation of this pathway may be of therapeutic interest. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  8. DCT protects human melanocytic cells from UVR and ROS damage and increases cell viability.

    Science.gov (United States)

    Ainger, Stephen A; Yong, Xuan L; Wong, Shu S; Skalamera, Dubravka; Gabrielli, Brian; Leonard, J Helen; Sturm, Richard A

    2014-12-01

    Dopachrome tautomerase (DCT) is involved in the formation of the photoprotective skin pigment eumelanin and has also been shown to have a role in response to apoptotic stimuli and oxidative stress. The effect of DCT on UVR DNA damage responses and survival pathways in human melanocytic cells was examined by knockdown experiments using melanoma cells, neonatal foreskin melanoblasts (MB) in monoculture and in co-culture with human keratinocytes. MB cell strains genotyped as either MC1R WT or MC1R RHC homozygotes, which are known to be deficient in DCT, were transduced with lentivirus vectors for either DCT knockdown or overexpression. We found melanoma cell survival was reduced by DCT depletion and by UVR over time. UVR-induced p53 and pp53-Ser15 levels were reduced with DCT depletion. Knockdown of DCT in MC1R WT and MC1R RHC MB cells reduced their survival after UVR exposure, whereas increased DCT protein levels enhanced survival. DCT depletion reduced p53 and pp53-Ser15 levels in WM266-4 melanoma and MC1R WT MB cells, while MC1R RHC MB cells displayed variable levels. Both MC1R WT and RHC genotypes of MB cells were responsive to UVR at 3 h with increases in both p53 and pp53-Ser15 proteins. MC1R WT MB cell strains in coculture with keratinocytes have an increased cell survival after UVR exposure when compared to those in monoculture, a protective effect which appears to be conferred by the keratinocytes.

  9. Ultraviolet stimulated melanogenesis by human melanocytes is augmented by di-acyl glycerol but not TPA

    Energy Technology Data Exchange (ETDEWEB)

    Friedmann, P.S.; Wren, F.E.; Matthews, J.N. (Univ. of Newcastle upon Tyne (England))

    1990-02-01

    Epidermal melanocytes (MC) synthesize melanin in response to ultraviolet radiation (UVR). The mechanisms mediating the UV-induced activation of melanogenesis are unknown but since UVR induces turnover of membrane phospholipids generating prostaglandins (PGs) and other products, it is possible that one of these might provide the activating signal. We have examined the effects of prostaglandins (PGs) E1, E2, D2, F2 alpha, and di-acyl glycerol upon the UV-induced responses of cultured human MC and the Cloudman S91 melanoma cell line. The PGs had little effect on unirradiated cells and did not alter the response to UVR in either human MC or S91 melanoma cells. However, a synthetic analogue of di-acyl glycerol, 1-oleyl 2-acetyl glycerol (OAG), caused a significant (P less than 0.0001), dose-related augmentation of melanin content both in human MC (seven-fold) and S91 cells (three-fold). UVR caused a significant augmentation of the OAG-induced melanogenesis of both human MC and S91 cells. Since OAG is known to activate protein kinase C, it was possible that the observed modulation of the UVR signal could be via that pathway. Di-octanoyl glycerol, another di-acyl glycerol, which activates kinase C, caused a small (70%) increase in melanogenesis in MC which was not altered by UVR. However, 12-0 tetradecanoyl phorbol 13-acetate (TPA), a potent activator of protein kinase C, had no significant effect on either basal or UV-induced melanin synthesis in either cell type. These data suggest that the UV-induced signal activating melanogenesis could be mediated by di-acyl glycerol. Furthermore, they imply that the signal is transduced via an alternative, pathway that might be independent of protein kinase C.

  10. Proteinase-activated receptor-2 stimulates prostaglandin production in keratinocytes: analysis of prostaglandin receptors on human melanocytes and effects of PGE2 and PGF2alpha on melanocyte dendricity.

    Science.gov (United States)

    Scott, Glynis; Leopardi, Sonya; Printup, Stacey; Malhi, Namrita; Seiberg, Miri; Lapoint, Randi

    2004-05-01

    Prostaglandins (PG) are key mediators of diverse functions in the skin and several reports suggest that PG mediate post-inflammatory pigmentary changes through modulation of melanocyte dendricity and melanin synthesis. The proteinase-activated receptor 2 (PAR-2) is important for skin pigmentation because activation of keratinocyte PAR-2 stimulates uptake of melanosomes through phagocytosis in a Rho-dependent manner. In this report, we show that activation of keratinocyte PAR-2 stimulates release of PGE(2) and PGF(2alpha) and that PGE(2) and PGF(2alpha) act as paracrine factors that stimulate melanocyte dendricity. We characterized the expression of the EP and FP receptors in human melanocytes and show that human melanocytes express EP1 and EP3, and the FP receptor, but not EP2 and EP4. Treatment of melanocytes with EP1 and EP3 receptor agonists resulted in increased melanocyte dendricity, indicating that both EP1 and EP3 receptor signaling contribute to PGE(2)-mediated melanocyte dendricity. Certain EP3 receptor subtypes have been shown to increase adenosine 3',5'-cyclic monophosphate (cAMP) through coupling to Gs, whereas EP1 is known to couple to Gq to activate phospholipase C with elevation in Ca(2+). The cAMP/protein kinase A system is known to modulate melanocyte dendrite formation through modulation of Rac and Rho activity. Neither PGF(2alpha) or PGE(2) elevated cAMP in human melanocytes showing that dendricity observed in response to PGE(2) and PGF(2alpha) is cAMP-independent. Our data suggest that PAR-2 mediates cutaneous pigmentation both through increased uptake of melanosomes by keratinocytes, as well as by release of PGE(2) and PGF(2alpha) that stimulate melanocyte dendricity through EP1, EP3, and FP receptors.

  11. 白细胞介素-22对人表皮黑素细胞生物活性的影响%Effects of IL-22 on biological activity of human epidermal melanocytes

    Institute of Scientific and Technical Information of China (English)

    王芳; 林新瑜; 刘刚; 廖恒利; 陈明懿; 刘伟; 王艺淳

    2014-01-01

    Objective To observe the effects of IL-22 on biological activity of human epidermal melanocytes, including proliferation, tyrosinase activity, and apoptosis of melanocytes. The results of the study would help us to better understand the pathogenesis of pigment aplastic skin diseases and provide new ideas for the treatment of pigment disorders. Methods An in vitro culture system of melanocytes simulated normal physiological state was established, then the effects of IL-22 of different concentrations on proliferative activity of human epidermal melanocytes were detected by CCK-8 method, meanwhile the effects on tyrosinase activity were detected by L-DOPA method, and the effects on apoptosis were detected by lfow cytometry. Results With the increase of concentration, IL-22 exhibited time-dependent inhibitory effect on the proliferation of melanocytes, for the inhibitory ratio of melanocytes, there was signiifcant difference between the IL-22 groups and the control group (P<0.05). For the melanocytes cultured at the concentration of 100 ng/ml of IL-22 and the control group, the apoptosis rate of melanocytes were 23.0%and 5.1%respectively after 48 h of cell culture. Conclusions With the increase of concentration, IL-22 exhibited time-dependent inhibitory effect on the proliferation of melanocytes. When the culture time over 72 hours, the proliferation of melanocytes was inhibited in IL-22 groups with all the different concentrations; meanwhile with the increase of the concentration of IL-22, the tyrosinase activity of melanocytes gradually decreased;compared with the control group, the apoptosis rate of melanocytes in IL-22 groups increased signiifcantly. All these results suggest that IL-22 may cause damage of melanocytes.%目的:观察白细胞介素(IL)-22对黑素细胞生物活性的影响,包括黑素细胞的增生、黑素细胞酪氨酸酶活性和黑素细胞凋亡,从而研究色素障碍性皮肤病的发病机制,为色素性疾病的治疗提供

  12. Effect of Q-switched Alexandrite laser irradiation on epidermal melanocytes in treatment of Nevus of Ota

    Institute of Scientific and Technical Information of China (English)

    卢忠; 陈军庞; 王侠生; 方丽华; 焦圣; 黄雯

    2003-01-01

    Objective To investigate injury to epidermal melanocyte by Q-switched Alexandrite laser.Methods Multiple biopsies were performed on 5 patients with nevus of Ota from before irradiation to 1 year after irradiation. Fourteen specimens were obtained for light microscopy, and 17 for transmission electron microscopy.Results Melanosomes in epidermal melanocytes were both smaller in size and fewer in number than those in dermal melanocytes. Immediately after irradiation, focal extracellular vacuoles of the basal layer could be observed under light microscopy. Most epidermal melanocytes underwent mild or moderate injury in the form of vacuolated melanosomes, swollen mitochondria, dilation of endoplasmic reticulum, and expansion of extracellular space, retaining intact cell membranes. Normal structures were restored 5 months to 1 year after irradiation, with no depigmentation or hyperpigmentation as seen by light microscopy.Conclusion Injury of melanosomes in epidermal melanocytes is reversible.

  13. Molecular cytogenetics of cutaneous melanocytic lesions - diagnostic, prognostic and therapeutic aspects.

    Science.gov (United States)

    Blokx, Willeke Am M; van Dijk, Marcory C R F; Ruiter, Dirk J

    2010-01-01

    This review intends to update current knowledge regarding molecular cytogenetics in melanocytic tumours with a focus on cutaneous melanocytic lesions. Advantages and limitations of diverse, already established methods, such as (fluorescence) in situ hybridization and mutation analysis, to detect these cytogenetic alterations in melanocytic tumours are described. In addition, the potential value of more novel techniques such as multiplex ligation-dependent probe amplification is pointed out. This review demonstrates that at present cytogenetics has mainly increased our understanding of the pathogenesis of melanocytic tumours, with an important role for activation of the mitogen-activated protein kinase (MAPK) signalling pathway in the initiation of melanocytic tumours. Mutations in BRAF (in common naevocellular naevi), NRAS (congenital naevi), HRAS (Spitz naevi) and GNAQ (blue naevi) can all cause MAPK activation. All these mutations seem early events in the development of melanocytic tumours, but by themselves are insufficient to cause progression towards melanoma. Additional molecular alterations are implicated in progression towards melanoma, with different genetic alterations in melanomas at different sites and with varying levels of sun exposure. This genetic heterogeneity in distinct types of naevi and melanomas can be used for the development of molecular tests for diagnostic purposes. However, at the moment only few molecular tests have become of diagnostic value and are performed in daily routine practice. This is caused by lack of large prospective studies on the diagnostic value of molecular tests including follow-up, and by the low prevalence of certain molecular alterations. For the future we foresee an increasing role for cytogenetics in the treatment of melanoma patients with the increasing availability of targeted therapy. Potential targets for metastatic melanoma include genes involved in the MAPK pathway, such as BRAF and RAS. More recently, KIT

  14. Porcine melanotic cutaneous lesions and lymph nodes: immunohistochemical differentiation of melanocytes and melanophages.

    Science.gov (United States)

    Grossi, A B; Hyttel, P; Jensen, H E; Leifsson, P S

    2015-01-01

    Porcine melanomas have proven interesting in a wider biological perspective due to a common phenomenon of spontaneous regression, which is characterized by infiltration of macrophages, among others. Separation of neoplastic melanocytes from pigment-laden macrophages may, however, be challenging as the morphology of melanocytes varies considerably and sometimes resembles macrophages. The aim of this study was correspondingly to characterize and differentiate the cells in 20 porcine melanocytomas and regional lymph nodes by histologic examination and immunohistochemistry for melan A, PNL2, S100, lysozyme, alpha-1-antitrypsin, and ionized calcium binding adaptor molecule 1 (Iba1). Grossly, the melanocytomas were divided into 2 distinct types: pigmented maculae (n = 7) and raised tumors (n = 13). In the maculae, the pigmented cells were mainly melanocytes reactive for melan A, PNL2 and S100. In contrast, the majority of the cells in the raised tumors were melanophages, which expressed Iba1, alpha-1-antitrypsin, and lysozyme. Yet, cells histomorphologically indistinguishable from the melanophages expressed melan A and PNL2. These cells were Iba1 and S100 negative, and ultrastructurally, they were devoid of lysosomal bodies and filled with stage III and IV melanosomes. In the regional lymph nodes, melanocytes were present in the trabecular sinuses. In focally or diffusely black lymph nodes, pigmentation was, however, mainly due to aggregates of melanophages, which were confined to the trabeculae, deep cortex, and peripheral lymphoreticular tissue. Normal and neoplastic porcine melanocytes express melan A and PNL2, and immunohistochemical staining for melan A, PNL2, and Iba1 was found useful to identify and distinguish melanocytes and melanophages in porcine melanotic lesions.

  15. Melanocytes and melanin represent a first line of innate immunity against Candida albicans.

    Science.gov (United States)

    Tapia, Cecilia V; Falconer, Maryanne; Tempio, Fabián; Falcón, Felipe; López, Mercedes; Fuentes, Marisol; Alburquenque, Claudio; Amaro, José; Bucarey, Sergio A; Di Nardo, Anna

    2014-07-01

    Melanocytes are dendritic cells located in the skin and mucosae that synthesize melanin. Some infections induce hypo- or hyperpigmentation, which is associated with the activation of Toll-like receptors (TLRs), especially TLR4. Candida albicans is an opportunist pathogen that can switch between blastoconidia and hyphae forms; the latter is associated with invasion. Our objectives in this study were to ascertain whether C. albicans induces pigmentation in melanocytes and whether this process is dependent on TLR activation, as well as relating this with the antifungal activity of melanin as a first line of innate immunity against fungal infections. Normal human melanocytes were stimulated with C. albicans supernatants or with crude extracts of the blastoconidia or hyphae forms, and pigmentation and TLR2/TLR4 expression were measured. Expression of the melanosomal antigens Melan-A and gp100 was examined for any correlation with increased melanin levels or antifungal activity in melanocyte lysates. Melanosomal antigens were induced earlier than cell pigmentation, and hyphae induced stronger melanization than blastoconidia. Notably, when melanocytes were stimulated with crude extracts of C. albicans, the cell surface expression of TLR2/TLR4 began at 48 h post-stimulation and peaked at 72 h. At this time, blastoconidia induced both TLR2 and TLR4 expression, whereas hyphae only induced TLR4 expression. Taken together, these results suggest that melanocytes play a key role in innate immune responses against C. albicans infections by recognizing pathogenic forms of C. albicans via TLR4, resulting in increased melanin content and inhibition of infection.

  16. Mechanisms contributing to differential regulation of PAX3 downstream target genes in normal human epidermal melanocytes versus melanoma cells.

    Science.gov (United States)

    Bartlett, Danielle; Boyle, Glen M; Ziman, Mel; Medic, Sandra

    2015-01-01

    Melanoma is a highly aggressive and drug resistant form of skin cancer. It arises from melanocytes, the pigment producing cells of the skin. The formation of these melanocytes is driven by the transcription factor PAX3 early during embryonic development. As a result of alternative splicing, the PAX3 gene gives rise to eight different transcripts which encode isoforms that have different structures and activate different downstream target genes involved in pathways of cell proliferation, migration, differentiation and survival. Furthermore, post-translational modifications have also been shown to alter the functions of PAX3. We previously identified PAX3 downstream target genes in melanocytes and melanoma cells. Here we assessed the effects of PAX3 down-regulation on this panel of target genes in primary melanocytes versus melanoma cells. We show that PAX3 differentially regulates various downstream target genes involved in cell proliferation in melanoma cells compared to melanocytes. To determine mechanisms behind this differential downstream target gene regulation, we performed immunoprecipitation to assess post-translational modifications of the PAX3 protein as well as RNAseq to determine PAX3 transcript expression profiles in melanocytes compared to melanoma cells. Although PAX3 was found to be post-translationally modified, there was no qualitative difference in phosphorylation and ubiquitination between melanocytes and melanoma cells, while acetylation of PAX3 was reduced in melanoma cells. Additionally, there were differences in PAX3 transcript expression profiles between melanocytes and melanoma cells. In particular the PAX3E transcript, responsible for reducing melanocyte proliferation and increasing apoptosis, was found to be down-regulated in melanoma cells compared to melanocytes. These results suggest that alternate transcript expression profiles activate different downstream target genes leading to the melanoma phenotype.

  17. Postnatal lineage mapping of follicular melanocytes with the Tyr∷CreERT2 transgene

    OpenAIRE

    Harris, Melissa L.; William J Pavan

    2012-01-01

    One of the main advantages of using inducible and conditional transgenes to study pigment cell biology is that they allow for genetic manipulation within melanocytes after roles in general neural crest or melanoblast development have been fulfilled. Specifically, we focus here on the ability of the Tyr∷CreERT2 transgenic line to alter genes within follicular melanocytes postnatally. Using the Gt(ROSA)26Sortm1sor reporter allele, we present in detail the expression and activity of Tyr∷CreERT2 ...

  18. 黑素细胞对同种异体淋巴细胞转化增殖的影响%The effects of Melanocyte to the Transormation and Proliferation of Allogeneic Lymphocyte

    Institute of Scientific and Technical Information of China (English)

    项蕾红; 郑志忠; 陈伟华; 祝绿川; 廖康煌

    2001-01-01

    目的观察体外培养人黑素细胞与同种异体的淋巴细胞混合后,黑素细胞对淋巴细胞的促转化、增殖作用。方法用3H-TdR掺入同位素液态闪烁计数法测定淋巴细胞的转化增殖率。实验结果应用t检验进行统计学分析。结果黑素细胞对同种异体淋巴细胞的转化增殖率与刀豆蛋白(conA)刺激淋巴细胞转化、增殖的阳性对照比较,结果显示黑素细胞的促淋巴细胞转化、增殖的特异性抗原作用较弱。进一步比较黑素细胞对不同病期的白癜风患者的淋巴细胞的影响,发现黑素细胞对活动期白癜风患者淋巴细胞的刺激作用相对较强,而稳定期患者和正常人对照组结果无显著性差异。结论正常人黑素细胞的特异性抗原作用较弱,为同种异体黑素细胞移植治疗白癜风创造了有利的条件。%Objective To observe the cultured melanocyte,as specificantigen,how to affect the transormation and proliferation of allogeneic lymphocytes in the mixed melanocyte lymphocyte reaction.Methods 3H-thymicine-added uptake was evaluated by means of liquid scintillation counting and expressed as cpm.The Results of lymphocyte proliferation were expressed by the stimulation indexes.The indexes of each group was analyzed statistically by t-test.Results The stimulation indexes of active vitiligo was significantly different compared with that of stable vitiligo and normal controls.The stimulation indexes of the group which melanocyte was used as specific antigen,was also significantly different compared with that of positive controls.Conclusion The melanocyte worked as specific antigen in mixed melanocyte lymphocyte reaction and its effect was weak.In the melanocyte allograft,the patients with stable vitiligo were the first choice.

  19. 白介素-10,白介素-17对人表皮黑素细胞生物活性的影响%Effects of IL-10, IL-17 on biological activity of human epidermal melanocytes

    Institute of Scientific and Technical Information of China (English)

    王芳; 林新瑜; 刘刚; 廖恒利; 刘伟; 陈明懿; 王艺淳

    2015-01-01

    melanocytes, the activity of tyrosinase were analyzed.Results ① With the increased culture time, the proliferation rate of cultured human epidermal melanocytes in the presence of IL-10 at the concentration of 50μg/mL increased.When melanocytes were cultured for 48h and 72h, there were statistically significant differences ( P<0.01) in proliferation rate of cultured human epidermal melanocytes compared with the control group.However, with the higher IL-10 concentration the proliferation rate of melanocytes manifests a downward trend with the prolonged in-cubation time.When melanocytes were cultured for 48h and 72h, the concentration rate of melanocytes in the pres-ence of IL-10 at the concentration of 200ng/ml has significant differences compared with the control group ( P<0.05) .②With the increased incubation time, the inhibitory ratio of melanocytes was increased in a dose-dependent manner in the presence of IL-17.The inhibitory ratio of melanocytes between the IL-17 groups and the control group were significant difference ( P<0.05) at the concentration of 50μg/ml and statistically significant ( P<0.01) at the concentration of 100μg/ml and 200μg/ml for 48h and 72h.③The melanocyte tyrosinase activity of IL-10 groups in-creased and there are significant differences between IL-10 groups and the control group at the concentration of 50μg/ml for 48h ( P<0.05) .The melanocyte tyrosinase activity were inhibited in the IL-17 group, and there were significant differences between the three groups and the control group at the concentration of 50μg/ml, 100μg/ml and 200μg/ml for the IL-17 group.④Melanocytes were cultured at the concentration of 100μg/ml of IL-10, IL-17 and control group for 48h, the apoptosis rate of melanocytes were 5.2%, 32.1%and 5.1%respectivel.Conclusion The appropriate concentration of IL-10 would promote the proliferation of melanocytes and the tyrosinase activity.IL-17 induced injuries on melanocytes, but the mechanisms need further research and

  20. Relativity of pure states entanglement

    CERN Document Server

    Zyczkowski, K; Zyczkowski, Karol; Bengtsson, Ingemar

    2002-01-01

    Entanglement of any pure state of an N times N bi-partite quantum system may be characterized by the vector of coefficients arising by its Schmidt decomposition. We analyze various measures of entanglement derived from the generalized entropies of the vector of Schmidt coefficients. For N >= 3 they generate different ordering in the set of pure states and for some states their ordering depends on the measure of entanglement used. This odd-looking property is acceptable, since these incomparable states cannot be transformed to each other with unit efficiency by any local operation. In analogy to special relativity the set of pure states equivalent under local unitaries has a causal structure so that at each point the set splits into three parts: the 'Future', the 'Past' and the set of noncomparable states.

  1. "Pure" cutaneous histiocytosis-X.

    Science.gov (United States)

    Wolfson, S L; Botero, F; Hurwitz, S; Pearson, H A

    1981-11-15

    The case histories of two young children who experienced skin rashes involving various areas of the body are reported. The diagnosis of pure cutaneous histiocytosis-X was established after extensive studies revealed no other organ involvement. The patients were treated with oral corticosteroids. Currently, both children are in good health, show no evidence of disease, and have been followed over a four-to-five-year period. Therapy with corticosteroids may not be indicated with pure cutaneous histiocytosis-X unless there is evidence of extracutaneous dissemination or rapid progression of the disease.

  2. Pure Spinors for General Backgrounds

    CERN Document Server

    Fre', Pietro

    2008-01-01

    We show the equivalence of the different types of pure spinor constraints geometrically derived from the Free Differential Algebras of N=2 d=10 supergravities. Firstly, we compute the general solutions of these constraints, using both a G_2 and an SO(8) covariant decomposition of the 10d chiral spinors. Secondly, we verify that the number of independent degrees of freedom is equal to that implied by the Poincare' pure spinor constraints so-far used for superstrings, namely twenty two. Thirdly, we show the equivalence between the FDA type IIA/B constraints among each other and with the Poincare' ones.

  3. sAPP as a regulator of dendrite motility and melanin release in epidermal melanocytes and melanoma cells.

    Science.gov (United States)

    Quast, Thomas; Wehner, Sven; Kirfel, Gregor; Jaeger, Klaus; De Luca, Michele; Herzog, Volker

    2003-09-01

    Numerous factors including ultraviolet (UV) radiation and growth factors regulate the specific function of epidermal melanocytes. A recently discovered epidermal growth factor is sAPP, the soluble N-terminal ectodomain of the beta-amyloid precursor protein (APP). Using whole mount preparations of isolated human epidermis, we detected a small population of basal cells, which expressed exceptionally high levels of APP. These cells were identified as melanocytes, which, similar to keratinocytes and neuronal cells, expressed the three APP isoforms 695, 751, and 770. They differed in their expression pattern from that of neuronal cells by expressing only low levels of APP 695. Melanocytes and melanoma cells in vitro released, in addition to keratinocytes, large quantities of sAPP. Because of its growth factor function, we studied possible effects of sAPP on melanocytes. Recombinant sAPP strongly increased lamellipodia activity at dendritic tips, an effect that coincided with increased release of melanin particles. Our observations point to the possible use of APP as an immunocytochemical marker for melanocytes. They suggest that sAPP derived from keratinocytes and/or melanocytes belongs to a family of factors operating in the paracrine and/or autocrine regulation of melanocyte function.

  4. The effect of bamboo (Phyllostachys nigra var. henenis Strapf) leaf extract on epidermal melanocytes in ultraviolet B-irradiated mice

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hae June; Chae, Se Lim; Kim, Sung Ho [Chonnam National Univ., Gwangju (Korea, Republic of)

    2007-06-15

    We induced the activation of melanocytes in the epidermis of C57BL/6 mice by ultraviolet B (UVB) irradiation and observed the effect of bamboo (phyllostachys nigra var. henenis strapf) Leaf Extract (BLE) on the formation, and decrease of UVB-induced epidermal melanocytes. C57BL/6 mice were irradiated by UVB 80 mJ/cm{sup 2} (0.5 mW/sec) daily for 7 days, and BLE was intraperitoneally or topically applied pre- or post-irradiation. For the estimation of change of epidermal melanocytes, light microscopic observation with DihydrOxyPhenylAlanine (DOPA) stain was performed. Split epidermal sheets prepared from the ear of untreated mice exhibited 11-16 melanocytes/mm{sup 2}, and one week after UV irradiation, the applied areas show an increased number of strongly DOPA-positive melanocytes with stout dendrites. But intraperitoneal or topical treatment with BLE before each irradiation interrupted UVB-induced pigmentation and resulted in a marked reduction in the number of epidermal melanocytes as compared to radiation control skin. The number and size of DOPA-positive epidermal melanocytes were also significantly decreased in intraperitoneally injected or topically applicated group after irradiation with BLE at 3rd and 6th weeks after irradiation. The results of present study indicate that BLE is likely to be useful as inhibitor of UVB-induced pigmentation and depigmenting agent.

  5. The effect of Bu-Zhong-Yi-Qi-Tang on epidermal melanocytes in ultraviolet B-irradiated mice

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hae June [Korea Institute of Radiological and Medical Science, Seoul (Korea, Republic of); Kim, Hwan Sung; Park, Young Jong; Kim, Joong Sun; Moon, Chang Jong; Kim, Jong Choon; Bae, Chun Sik; Kim, Sung Ho [Chonnam National University, Gwangju (Korea, Republic of); Jo, Sung Kee [KAERI, Daejeon (Korea, Republic of)

    2008-09-15

    We induced the activation of melanocytes in the epidermis of C57BL/6 mice by UltraViolet B (UVB) irradiation and observed the effect of Bu-Zhong-Yi-Qi-Tang (BZYQT) on the formation, and decrease of UVB-induced epidermal melanocytes. C57BL/6 mice were irradiated by UVB 80 mJ/cm{sup 2} (0.5 mW/sec) daily for 7 days, and BZYQT was intraperitoneally or topically applied pre- or post-irradiation. For the estimation of change of epidermal melanocytes, light microscopic observation with dihydroxyphenylalanine (DOPA) stain was performed. Split epidermal sheets prepared from the ear of untreated mice exhibited 11-16 melanocytes/mm{sup 2}, and one week after UV irradiation, the applied areas show an increased number of strongly DOPA-positive melanocytes with stout dendrites. But intraperitoneal or topical treatment with BZYQT before each irradiation interrupted UVB-induced pigmentation and resulted in a marked reduction in the number of epidermal melanocytes as compared to radiation control skin. The number and size of DOPA-positive epidermal melanocytes were also significantly decreased in intraperitoneally injected or topically applicated group after irradiation with BZYQT at 3rd and 6th weeks after irradiation. The present study suggests the BZYQT as inhibitor of UVB-induced pigmentation and depigmenting agent.

  6. The unfolded protein response in melanocytes: activation in response to chemical stressors of the endoplasmic reticulum and tyrosinase misfolding.

    Science.gov (United States)

    Manga, Prashiela; Bis, Sabina; Knoll, Kristen; Perez, Beremis; Orlow, Seth J

    2010-10-01

    Accumulation of proteins in the endoplasmic reticulum (ER) triggers the unfolded protein response (UPR), comprising three signaling pathways initiated by Ire1, Perk and Atf6 respectively. Unfolded protein response activation was compared in chemically stressed murine wildtype melanocytes and mutant melanocytes that retain tyrosinase in the ER. Thapsigargin, an ER stressor, activated all pathways in wildtype melanocytes, triggering Caspase 12-mediated apoptosis at toxic doses. Albino melanocytes expressing mutant tyrosinase showed evidence of ER stress with increased Ire1 expression, but the downstream effector, Xbp1, was not activated even following thapsigargin treatment. Attenuation of Ire1 signaling was recapitulated in wildtype melanocytes treated with thapsigargin for 8 days, with diminished Xbp1 activation observed after 4 days. Atf6 was also activated in albino melanocytes, with no response to thapsigargin, while the Perk pathway was not activated and thapsigargin treatment elicited robust expression of the downstream effector CCAAT-enhancer-binding protein homologous protein. Thus, melanocytes adapt to ER stress by attenuating two UPR pathways.

  7. The epidermal melanocyte system in individuals of Scandinavian origin, determined by DOPA-staining and TEM

    DEFF Research Database (Denmark)

    Drzewiecki, K T; Piltz-Drzewiecka, J

    1979-01-01

    in some specimens. This is due partly to the preparation procedure and partly to normal biological variations. We believe that we have demonstrated a cyclic function of the melanocyte in the epidermis. The varying density of cells in epidermal sheets as well as their varying morphology support the theory...

  8. Progressive Increase in Telomerase Activity From Benign Melanocytic Conditions to Malignant Melanoma

    Directory of Open Access Journals (Sweden)

    Ruben D. Ramirez

    1999-04-01

    Full Text Available The expression of telomerase activity and the in situ localization of the human telomerase RNA component (hTR in melanocytic skin lesions was evaluated in specimens from sixty-three patients. Specimens of melanocytic nevi, primary melanomas and subcutaneous metastases of melanoma were obtained from fifty-eight patients, whereas metastasized lymph nodes were obtained from five patients. Telomerase activity was determined in these specimens by using a Polymerase Chain Reaction—based assay (TRAP. High relative mean telomerase activity levels were detected in metastatic melanoma (subcutaneous metastasess = 54.5, lymph node metastasess = 56.5. Much lower levels were detected in primary melanomas, which increased with advancing levels of tumor cell penetration (Clark II = 0.02, Clark III = 1.1, and Clark IV = 1.9. Twenty-six formalin-fixed, paraffin-embedded melanocytic lesions were sectioned and analyzed for telomerase RNA with a radioactive in situ hybridization assay. In situ hybridization studies with a probe to the template RNA component of telomerase confirmed that expression was almost exclusively confined to tumor cells and not infiltrating lymphocytes. These results indicate that levels of telomerase activity and telomerase RNA in melanocytic lesions correlate well with clinical stage and could potentially assist in the diagnosis of borderline lesions.

  9. Ultraviolet Radiation and the Slug Transcription Factor Induce Proinflammatory and Immunomodulatory Mediator Expression in Melanocytes

    Directory of Open Access Journals (Sweden)

    Stephanie H. Shirley

    2012-01-01

    Full Text Available Despite extensive investigation, the precise contribution of the ultraviolet radiation (UVR component of sunlight to melanoma etiology remains unclear. UVR induces keratinocytes to secrete proinflammatory and immunomodulatory mediators that promote inflammation and skin tumor development; expression of the slug transcription factor in keratinocytes is required for maximal production of these mediators. In the present studies we examined the possibility that UVR-exposed melanocytes also produce proinflammatory mediators and that Slug is important in this process. Microarray studies revealed that both UVR exposure and Slug overexpression altered transcription of a variety of proinflammatory mediators by normal human melanocytes; some of these mediators are also known to stimulate melanocyte growth and migration. There was little overlap in the spectra of cytokines produced by the two stimuli. However IL-20 was similarly induced by both stimuli and the NFκB pathway appeared to be important in both circumstances. Further exploration of UVR-induced and Slug-dependent pathways of cytokine induction in melanocytes may reveal novel targets for melanoma therapy.

  10. Postnatal lineage mapping of follicular melanocytes with the Tyr::CreER(T) (2) transgene.

    Science.gov (United States)

    Harris, Melissa L; Pavan, William J

    2013-03-01

    One of the main advantages of using inducible and conditional transgenes to study pigment cell biology is that they allow for genetic manipulation within melanocytes after roles in general neural crest or melanoblast development have been fulfilled. Specifically, we focus here on the ability of the Tyr::CreER(T) (2) transgenic line to alter genes within follicular melanocytes postnatally. Using the Gt(ROSA)26Sor(tm1sor) reporter allele, we present in detail the expression and activity of Tyr::CreER(T) (2) when induced during hair morphogenesis and adult hair cycling. We find that despite similarities in expression pattern to endogenous TYR, Tyr::CreER(T) (2) is effective at targeting both undifferentiated and differentiated melanocytes within the hair follicle. We also find that Tyr::CreER(T) (2) provides the highest levels of recombination when induced during the early phases of hair growth. In conclusion, the descriptions provided here will guide future analyses of gene function within the melanocyte system of the hair follicle when using this Tyr::CreER(T) (2) transgene.

  11. Behandling og håndtering af kongenitte melanocytære naevi

    DEFF Research Database (Denmark)

    Bahn, Kamille-Amalie; Hædersdal, Merete; Schmidt, Grethe

    2016-01-01

    Congenital melanocytic naevi (CMN) appear in approximately 2.6% of Caucasians. There is a major demand for treatment but no vital indication. Laser therapy, curettage and excision are available treatment modalities, but there is no ideal treatment with documented long-term effect and without side...

  12. The immunology and inflammatory responses of human melanocytes in infectious diseases.

    Science.gov (United States)

    Gasque, Philippe; Jaffar-Bandjee, Marie Christine

    2015-10-01

    Melanin is a canonical and major defense molecule in invertebrates but its role in mammalian immunity remains unexplored. In contrast, several recent studies have highlighted the emerging innate immune activities of human melanin-producing cells which can sense and respond to bacterial and viral infections. Indeed, the skin is a major portal of entry for pathogens such as arboviruses (Chikungunya, Dengue) and bacteria (mycobacterium leprae, Leptospira spirochetes). Melanocytes of the epidermis could contribute to the phagocytosis of these invading pathogens and to present antigens to competent immune cells. Melanocytes are known to produce key cytokines such as IL-1β, IL6 and TNF-α as well as chemokines. These molecules will subsequently alert macrophages, neutrophils, fibroblasts and keratinocytes through unique crosstalk mechanisms. The infection and the inflammatory responses will control melanocyte's immune and metabolic functions and could contribute to skin manifestations (rash, hyper or de-pigmentation, epidermolysis and psoriasis-like lesions). This review will address the potential role of melanocytes in immunity, inflammation and infection of the skin in health and diseases.

  13. Expert review remains important in the histopathological diagnosis of cutaneous melanocytic lesions

    NARCIS (Netherlands)

    van Dijk, M. C. R. F.; Aben, K. K. H.; van Hees, F.; Klaasen, A.; Blokx, W. A. M.; Kiemeney, L. A. L. M.; Ruiter, D. J.

    2008-01-01

    Aims: To assess the type of problems encountered in diagnosing melanocytic lesions and to evaluate the contribution of expert review. Methods and results: Data from 1887 lesions submitted for consultation to one of the expert pathologists of the Dutch Melanoma Working Group Pathology Panel between 1

  14. Disruption of melanosome transport in melanocytes treated with theophylline causes their degradation by autophagy.

    Science.gov (United States)

    Katsuyama, Yushi; Taira, Norihisa; Yoshioka, Masato; Okano, Yuri; Masaki, Hitoshi

    2017-02-07

    Melanosomes containing melanin are transported from the perinuclear area to the tips of dendrites in epidermal melanocytes, and are then transferred to keratinocytes. Thus, skin color is determined by the amount of melanin synthesized in melanocytes and the subsequent dispersion of melanosomes in the epidermis. Therefore, disrupting intracellular melanosome transport in melanocytes is considered an effective approach to regulate skin color. However, the fate of melanosomes that accumulate in melanocytes due to disrupted intracellular transport is unclear. In this study, we disrupted melanosome transport by knockdown of the motor protein MyosinVa. Knock-down of MyosinVa (M-KD) in cells treated with theophylline significantly down-regulated the mRNA and protein expression levels of tyrosinase. Interestingly, intracellular melanin contents in M-KD cells were decreased. Furthermore, M-KD cells showed activation of autophagy through increased expression of Microtubule-associated protein 1 light chain 3 (LC3) -II and decreased expression of p62. The sum of these results indicate that disruption of melanosome transport causes their degradation by autophagy.

  15. Uncovering the Role of BMP Signaling in Melanocyte Development and Melanoma Tumorigenesis

    Science.gov (United States)

    2014-07-01

    generated using TALEN -mediated mutagenesis3, transfections into human primed melanocytes and melanoma cells performed, and lines stably expressing GDF6...Cermak, E.J. Doyle, et al., Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Nuc. Acids Res. 39

  16. Enhanced bleaching treatment: opportunities for immune-assisted melanocyte suicide in vitiligo.

    Science.gov (United States)

    Webb, Kirsten C; Eby, Jonathan M; Hariharan, Vidhya; Hernandez, Claudia; Luiten, Rosalie M; Le Poole, I Caroline

    2014-08-01

    Depigmentation in vitiligo occurs by progressive loss of melanocytes from the basal layer of the skin, and can be psychologically devastating to patients. T cell-mediated autoimmunity explains the progressive nature of this disease. Rather than being confronted with periods of rapid depigmentation and bouts of repigmentation, patients with long-standing, treatment-resistant vitiligo can undergo depigmentation treatment. The objective is to remove residual pigmentation to achieve a cosmetically acceptable result--that of skin with a uniform appearance. In the United States, only the use of mono-benzyl ether of hydroquinone (MBEH) is approved for this purpose. However, satisfactory results can take time to appear, and there is a risk of repigmentation. MBEH induces necrotic melanocyte death followed by a cytotoxic T-cell response to remaining, distant melanocytes. As cytotoxic T-cell responses are instrumental to depigmentation, we propose that combining MBEH with immune adjuvant therapies will accelerate immune-mediated melanocyte destruction to achieve faster, more definitive depigmentation than with MBEH alone. As Toll-like Receptor (TLR) agonists--imiquimod, CpG, and Heat Shock Protein 70 (HSP 70)--all support powerful Th1 responses, we propose that using MBEH in combination with these agents can achieve superior depigmentation results for vitiligo patients.

  17. Induction of eruptive benign melanocytic naevi by immune suppressive agents, including biologicals.

    NARCIS (Netherlands)

    Bovenschen, H.J.; Tjioe, M.; Vermaat, H.; Hoop, D. de; Witteman, B.M.; Janssens, R.W.A.; Stoof, T.J.; Kerkhof, P.C.M. van de

    2006-01-01

    BACKGROUND: Eruptive naevi have been described to potentially arise in immune compromised patients. OBJECTIVES: We describe three patients with eruptive benign melanocytic naevi during a phase of immunosuppressive therapy. METHODS/DIAGNOSIS: Two patients with Crohn disease were treated with either a

  18. Molecular effects of 1-naphthyl-methylcarbamate and solar radiation exposures on human melanocytes.

    Science.gov (United States)

    Ferrucio, Bianca; Tiago, Manoela; Fannin, Richard D; Liu, Liwen; Gerrish, Kevin; Maria-Engler, Silvya Stuchi; Paules, Richard S; Barros, Silvia Berlanga de Moraes

    2017-02-01

    Carbaryl (1-naphthyl-methylcarbamate), a broad-spectrum insecticide, has recently been associated with the development of cutaneous melanoma in an epidemiological cohort study with U.S. farm workers also exposed to ultraviolet radiation, the main etiologic factor for skin carcinogenesis. We hypothesized that carbaryl exposure may increase deleterious effects of UV solar radiation on skin melanocytes. This study aimed to characterize human melanocytes after individual or combined exposure to carbaryl (100μM) and solar radiation (375mJ/cm(2)). In a microarray analysis, carbaryl, but not solar radiation, induced an oxidative stress response, evidenced by the upregulation of antioxidant genes, such as Hemeoxygenase-1 (HMOX1), and downregulation of Microphtalmia-associated Transcription Factor (MITF), the main regulator of melanocytic activity; results were confirmed by qRT-PCR. Carbaryl and solar radiation induced a gene response suggestive of DNA damage and cell cycle alteration. The expression of CDKN1A, BRCA1/2 and MDM2 genes was notably more intense in the combined treatment group, in a synergistic manner. Flow cytometry assays demonstrated S-phase cell cycle arrest, reduced apoptosis levels and faster induction of cyclobutane pyrimidine dimers (CPD) lesions in carbaryl treated groups. Our data suggests that carbaryl is genotoxic to human melanocytes, especially when associated with solar radiation.

  19. Development of a three-dimensional surface imaging system for melanocytic skin lesion evaluation

    Science.gov (United States)

    Tosca, Androniki; Kokolakis, Athanasios; Lasithiotakis, Konstantinos; Zacharopoulos, Athanasios; Zabulis, Xenophon; Marnelakis, Ioannis; Ripoll, Jorge; Stephanidis, Constantine

    2013-01-01

    Even though surface morphology is always taken into account when assessing clinically pigmented skin lesions, it is not captured by most modern imaging systems using digital imaging. Our aim is to develop a novel three-dimensional (3D) imaging technique to record detailed information of the surface anatomy of melanocytic lesions that will enable improved classification through digital imaging. The apparatus consists of three high-resolution cameras, a light source, and accompanying software. Volume measurements of specific phantoms using volumetric tubes render slightly lower values than those obtained by our 3D imaging system (mean%±SD, 3.8%±0.98, P<0.05). To examine the reproducibility of the method, sequential imaging of melanocytic lesions is carried out. The mean%±SD differences of area, major axis length, volume, and maximum height are 2.1%±1.1, 0.9%±0.8, 3.8%±2.9, and 2.5%±3.5, respectively. Thirty melanocytic lesions are assessed, including common and dysplastic nevi and melanomas. There is a significant difference between nevi and melanomas in terms of variance in height and boundary asymmetry (P<0.001). Moreover, dysplastic nevi have significantly higher variances in pigment density values than common nevi (P<0.001). Preliminary data suggest that our instrument has great potential in the evaluation of the melanocytic lesions. However, these findings should be confirmed in larger-scale studies.

  20. Clinical and histological responses of congenital melanocytic nevi after single treatment with Q-switched lasers

    NARCIS (Netherlands)

    Grevelink, JM; vanLeeuwen, RL; Anderson, RR; Byers, HR

    1997-01-01

    Background: Laser irradiation of congenital melanocytic nevi is a controversial treatment. Recurrence of lesions after laser treatment appears to be the rule, and the effects of laser irradiation on cellular biological behavior and the possible mutagenic responses of nevomelanocytes that have receiv

  1. Clinical and histological responses of congenital melanocytic nevi after single treatment with Q-switched lasers

    NARCIS (Netherlands)

    Grevelink, JM; vanLeeuwen, RL; Anderson, RR; Byers, HR

    1997-01-01

    Background: Laser irradiation of congenital melanocytic nevi is a controversial treatment. Recurrence of lesions after laser treatment appears to be the rule, and the effects of laser irradiation on cellular biological behavior and the possible mutagenic responses of nevomelanocytes that have receiv

  2. The epidermal melanocyte system in individuals of Scandinavian origin, determined by DOPA-staining and TEM

    DEFF Research Database (Denmark)

    Drzewiecki, K T; Piltz-Drzewiecka, J

    1979-01-01

    in some specimens. This is due partly to the preparation procedure and partly to normal biological variations. We believe that we have demonstrated a cyclic function of the melanocyte in the epidermis. The varying density of cells in epidermal sheets as well as their varying morphology support the theory...

  3. Discrimination of Dysplastic Nevi from Common Melanocytic Nevi by Cellular and Molecular Criteria.

    Science.gov (United States)

    Mitsui, Hiroshi; Kiecker, Felix; Shemer, Avner; Cannizzaro, Maria Vittoria; Wang, Claire Q F; Gulati, Nicholas; Ohmatsu, Hanako; Shah, Kejal R; Gilleaudeau, Patricia; Sullivan-Whalen, Mary; Cueto, Inna; McNutt, Neil Scott; Suárez-Fariñas, Mayte; Krueger, James G

    2016-10-01

    Dysplastic nevi (DNs), also known as Clark's nevi or atypical moles, are distinguished from common melanocytic nevi by variegation in pigmentation and clinical appearance, as well as differences in tissue patterning. However, cellular and molecular differences between DNs and common melanocytic nevi are not completely understood. Using cDNA microarray, quantitative RT-PCR, and immunohistochemistry, we molecularly characterized DNs and analyzed the difference between DNs and common melanocytic nevi. A total of 111 probesets (91 annotated genes, fold change > 2.0 and false discovery rate DNs was altered differentiation and activation of epidermal keratinocytes with increased expression of hair follicle-related molecules (keratin 25, trichohyalin, ribonuclease, RNase A family, 7) and inflammation-related molecules (S100A7, S100A8) at both genomic and protein levels. The immune microenvironment of DNs was characterized by an increase of T helper type 1 (IFNγ) and T helper type 2 (IL13) cytokines as well as an upregulation of oncostatin M and CXCL1. DUSP3, which regulates cellular senescence, was identified as one of the disease discriminative genes between DNs and common melanocytic nevi by three independent statistical approaches and its altered expression was confirmed by immunohistochemistry. The molecular and cellular changes in which the epidermal-melanin unit undergoes follicular differentiation as well as upregulation of defined cytokines could drive complex immune, epidermal, and pigmentary alterations.

  4. Pure robotic retrocaval ureter repair

    Directory of Open Access Journals (Sweden)

    Ashok k. Hemal

    2008-12-01

    Full Text Available PURPOSE: To demonstrate the feasibility of pure robotic retrocaval ureter repair. MATERIALS AND METHODS: A 33 year old female presented with right loin pain and obstruction on intravenous urography with the classical "fish-hook" appearance. She was counseled on the various methods of repair and elected to have a robot assisted repair. The following steps are performed during a pure robotic retrocaval ureter repair. The patient is placed in a modified flank position, pneumoperitoneum created and ports inserted. The colon is mobilized to expose the retroperitoneal structures: inferior vena cava, right gonadal vein, right ureter, and duodenum. The renal pelvis and ureter are mobilized and the renal pelvis transected. The ureter is transposed anterior to the inferior vena cava and a pyelopyelostomy is performed over a JJ stent. RESULTS: This patient was discharged on postoperative day 3. The catheter and drain tube were removed on day 1. Her JJ stent was removed at 6 weeks postoperatively. The postoperative intravenous urography at 3 months confirmed normal drainage of contrast medium. CONCLUSION: Pure robotic retrocaval ureter is a feasible procedure; however, there does not appear to be any great advantage over pure laparoscopy, apart from the ergonomic ease for the surgeon as well the simpler intracorporeal suturing.

  5. The usefulness of c-Kit in the immunohistochemical assessment of melanocytic lesions

    Science.gov (United States)

    Pilloni, L.; Bianco, P.; Difelice, E.; Cabras, S.; Castellanos, M.E.; Atzori, L.; Ferreli, C.; Mulas, P.; Nemolato, S.; Faa, G.

    2011-01-01

    C-Kit (CD117), the receptor for the stem cell factor, a growth factor for melanocyte migration and proliferation, has shown differential immunostaining in various benign and malignant melanocytic lesions. The purpose of this study is to compare c-Kit immunostaining in benign nevi and in primary and metastatic malignant melanomas, to determine whether c-Kit can aid in the differential diagnosis of these lesions. c-Kit immunostaining was performed in 60 cases of pigmented lesions, including 39 benign nevi (5 blue nevi, 5 intra-dermal nevi, 3 junctional nevi, 15 cases of primary compound nevus, 11 cases of Spitz nevus), 18 cases of primary malignant melanoma and 3 cases of metastatic melanoma. The vast majority of nevi and melanomas examined in this study were positive for c-Kit, with minimal differences between benign and malignant lesions. C-Kit cytoplasmatic immunoreactivity in the intraepidermal proliferating nevus cells, was detected in benign pigmented lesions as well as in malignant melanoma, increasing with the age of patients (P=0.007) in both groups. The patient’s age at presentation appeared to be the variable able to cluster benign and malignant pigmented lesions. The percentage of c-Kit positive intraepidermal nevus cells was better associated with age despite other variables (P=0.014). The intensity and percentage of c-Kit positivity in the proliferating nevus cells in the dermis was significantly increased in malignant melanocytic lesions (P=0.015 and P=0.008) compared to benign lesions (compound melanocytic nevi, Spitz nevi, intradermal nevi, blue nevi). Immunostaning for c-Kit in metastatic melanomas was negative. Interestingly in two cases of melanoma occurring on a pre-existent nevus, the melanoma tumor cells showed strong cytoplasmatic and membranous positivity for c-kit, in contrast with the absence of any immunoreactivity in pre-existent intradermal nevus cells. C-Kit does not appear to be a strong immunohistochemical marker for distinguishing

  6. Diagnosis and staging of female genital tract melanocytic lesions using pump-probe microscopy (Conference Presentation)

    Science.gov (United States)

    Robles, Francisco E.; Selim, Maria A.; Warren, Warren S.

    2016-02-01

    Melanoma of the vulva is the second most common type of malignancy afflicting that organ. This disease caries poor prognosis, and shows tendencies to recur locally and develop distant metastases through hematogenous dissemination. Further, there exists significant clinical overlap between early-stage melanomas and melanotic macules, benign lesions that are believed to develop in about 10% of the general female population. In this work we apply a novel nonlinear optical method, pump-probe microscopy, to quantitatively analyze female genitalia tract melanocytic lesions. Pump-probe microscopy provides chemical information of endogenous pigments by probing their electronic excited state dynamics, with subcellular resolution. Using unstained biopsy sections from 31 patients, we find significant differences between melanin type and structure in tissue regions with invasive melanoma, melanoma in-situ and non-malignant melanocytic proliferations (e.g., nevi, melanocytic macules). The molecular images of non-malignant lesion have a well-organized structure, with relatively homogenous pigment chemistry, most often consistent with that of eumelanin with large aggregate size or void of metals, such as iron. On the other hand, pigment type and structure observed in melanomas in-situ and invasive melanomas is typically much more heterogeneous, with larger contributions from pheomelanin, melanins with larger metal content, and/or melanins with smaller aggregate size. Of most significance, clear differences can be observed between melanocytic macules and vulvar melanoma in-situ, which, as discussed above, can be difficult to clinically distinguish. This initial study demonstrates pump-probe microscopy's potential as an adjuvant diagnostic tool by revealing systematic chemical and morphological differences in melanin pigmentation among invasive melanoma, melanoma in-situ and non-malignant melanocytic lesions.

  7. Effects of hypergravity on the expression of multidrug resistance proteins in human melanocytic cells

    Science.gov (United States)

    Lambers, B.; Stieber, C.; Grigorieva, O.; Block, I.; Bromeis, B.; Buravkova, L.; Gerzer, R.; Ivanova, K.

    In humans the skin serves as a barrier against potentially harmful effects of the environment Human melanocytes constitute the principal cells for skin pigmentation by synthesizing the pigment melanin Melanin acts as a scavenger for free radicals that may arise during metabolic stress The melanocytes are also able to secrete a wide range of signal molecules In previous studies we found that normal human melanocytes NHMs and non-metastatic melanoma cells respond to long-time exposure to hypergravity up to 5 g for 24 h with elevated efflux of guanosine 3 5 -cyclic monophosphate cGMP in the presence of phosphodiesterase PDE inhibitors e g 3-isobutyl-1-methylxanthine Cyclic GMP is known to play a signaling role in human melanocyte physiology It controls the signaling activities of nitric oxide NO in relation to melanogenesis as well as in melanocyte-extracellular matrix interactions that may be important for some pathological processes including metastasis The present study investigated the effects of hypergravity on the expression of the multidrug resistance proteins MRP 4 and 5 as highly selective cGMP exporters in non-stimulated and NO-stimulated NHMs and melanoma cells MCs on mRNA levels using semi-quantitative RT-PCR analysis Hypergravity up to 5 g for 24 h was produced by horizontal centrifugal acceleration The NONOate DETA-NO 0 1 mM was used as a direct NO donor for cell stimulation For 5-g experiments the mRNA levels for the highly specific cGMP transporter MRP5 appeared to be

  8. 羧甲基壳聚糖膜对人皮肤黑素细胞的生物相容性研究%Biocompatibility of carboxymethyl chitosan membranes with human skin melanocytes

    Institute of Scientific and Technical Information of China (English)

    孔玉龙; 王克玉; 张秀文; 马伟元

    2014-01-01

    Objective To study the biocompatibility of carboxymethyl chitosan (CMCS) membrane with melanocytes from healthy human skin,and to investigate the feasibility to transport and carry melanocytes by using CMCS membrane.Methods CMCS membrane was prepared by a casting method combined with a glutaraldehydebased cross-linking method.Melanocytes were isolated from the foreskin of healthy men,and subjected to primary culture and subculture.The third-passage melanocytes were classified into two groups to be cultured on the CMCS membrane (test group) or traditional culture plates (control group).Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferative activity of melanocytes,and a sodium hydroxide-based lysis method to determine melanin content.HMB45 staining was conducted,and tyrosinase activity was estimated for melanocytes.Results Inverted microscopy showed that melanocytes were evenly distributed on the CMCS membrane with a normal shape.The melanocytes adherent to the CMCS membrane stained positive for anti-HMB45 monoclonal antibody.The growth curve of the melanocytes on the CMCS membrane,which was obtained from MTT assay,demonstrated that CMCS membrane could support the normal growth of melanocytes.No significant difference was observed between the test group and control group in melanin content (0.083 ± 0.015 vs.0.066 ± 0.008,t =2.38,P > 0.01) or tyrosinase activity (0.234 ± 0.083 vs.0.241 ± 0.061,t =0.23,P > 0.05).Conclusion CMCS membrane can maintain the normal biological activity of melanocytes and have good biocompatibility with skin melanocytes.%目的 探讨羧甲基壳聚糖膜对健康人黑素细胞的生物相容性,探索羧甲基壳聚糖膜搭载黑素细胞的可行性.方法 使用流延法联合戊二醛交联法制备羧甲基壳聚糖膜,黑素细胞分离培养技术分离培养健康人黑素细胞,使用HMB45染色、MTT法、NaOH裂解法、酪氨酸酶活性检测等方法检测羧甲基壳

  9. Manifolds of interconvertible pure states

    OpenAIRE

    Sinolecka, Magdalena M.; Zyczkowski, Karol; Kus, Marek

    2001-01-01

    Local orbits of a pure state of an N x N bi-partite quantum system are analyzed. We compute their dimensions which depends on the degeneracy of the vector of coefficients arising by the Schmidt decomposition. In particular, the generic orbit has 2N^2 -N-1 dimensions, the set of separable states is 4(N-1) dimensional, while the manifold of maximally entangled states has N^2-1 dimensions.

  10. Manifolds of interconvertible pure states

    CERN Document Server

    Sinolecka, M M; Kus, M; Sinolecka, Magdalena M.; Zyczkowski, Karol; Kus, Marek

    2002-01-01

    Local orbits of a pure state of an N x N bi-partite quantum system are analyzed. We compute their dimensions which depends on the degeneracy of the vector of coefficients arising by the Schmidt decomposition. In particular, the generic orbit has 2N^2 -N-1 dimensions, the set of separable states is 4(N-1) dimensional, while the manifold of maximally entangled states has N^2-1 dimensions.

  11. Multimedia programming with pure data

    CERN Document Server

    Chung, Bryan

    2013-01-01

    A quick and comprehensive tutorial book for media designers to jump-start interactive multimedia production with computer graphics, digital audio, digital video, and interactivity, using the Pure Data graphical programming environment.An introductory book on multimedia programming for media artists/designers who like to work on interactivity in their projects, digital art/design students who like to learn the first multimedia programming technique, and audio-visual performers who like to customize their performance sets

  12. 超低温液氮冷冻技术冻存复苏黑素细胞移植治疗大面积白癜风%Ultra-low temperature liquid nitrogen cryotherapy cryopreservation melanocyte transplantment in the treatment of large area vitiligo

    Institute of Scientific and Technical Information of China (English)

    洪为松; 傅丽芳; 尉晓冬; 钱国培; 许爱娥

    2012-01-01

    目的:研究应用超低温液氮冷冻技术冻存复苏黑素细胞的生物学活性及分析自体黑素细胞移植治疗大面积白癜风的疗效.方法:负压吸疱获取患者正常表皮片,Hu16培养基体外培养黑素细胞,应用超低温液氮冷冻技术冻存复苏黑素细胞,检测黑素细胞的分裂时间(DOT)、黑素含量(M)、黑素制造量(MP)和树突数.受皮区应用超脉冲二氧化碳激光磨削去表皮后,以黑素细胞密度6~10x104个/cm2进行移植.所有患者均至少跟踪观察疗效6个月.结果:11例患者黑素细胞培养至第二代进行超低温液氮冷冻,15例患者黑素细胞培养至第三代进行冷冻,所有的26例患者冻存6、12和24个月复苏黑素细胞的生物学活性指标DOT、M、MP和树突数均与未冻存的细胞无明显差异.未冻存、冻存6、12和24个月的黑素细胞移植治疗的痊愈率为57.69%、61.54%、57.69%和53.85%,有效率为84.61%、88.46%、80.77%和80.77%.冻存6、12和24个月的黑素细胞移植的痊愈率和有效率与未冻存的黑素细胞移植的痊愈率和有效率均无显著性差异.结论:超低温液氮冷冻技术能很好的储存白癜风患者体外培养的黑素细胞,应用冻存复苏的黑素细胞自体移植治疗大面积白瘢风皮损疗效确切.%Objective To investigate the effect of ultra-low temperature liquid nitrogen cryotherapy cryopreservation for the biological activity of melanocytes and to evaluate the effectiveness of the transplantation of cultured autologous melanocyte (TCAM) in the treatment of large area lesion of vitiligo. Methods Autologous donor skin sheet was obtained by suction blisters from the normal skin of vitiligo patient. The isolated melanocytes were cultured in Hu16 media, and applied the ultra-low temperature liquid nitrogen cryotherapy cryopreservation melanocytes, the cell division time (DOT) and melanin content and the melanin manufacturing capacity and the number of

  13. Role of nitric oxide and cyclic GMP signaling in melanocyte response to hypergravity

    Science.gov (United States)

    Ivanova, Krassimira; Lambers, Britta; Tsiockas, Wasiliki; Block, Ingrid; Gerzer, Rupert

    Nitric oxide (NO) has a prominent role in many (patho)physiological processes in the skin including erythema, inflammation, and cancerogenesis. The soluble guanylyl cyclase (sGC), a key transducer in NO signaling, catalyzes the formation of the second messenger guanosine 3´,5´-cyclic monophosphate (cyclic cGMP or cGMP). For human melanocytes, which are responsible for skin pigmentation by synthesizing the pigment melanin, it has been reported that the NO/sGC/cGMP pathway is involved in UVB-induced melanogenesis. Melanin acts as a scavenger for free radicals that may arise during metabolic stress. It may also act as a photosensitizer that generates active oxygen species upon UV irradiation, which may initiate hypopigmentary disorders (e.g., vitiligo) as well as UV-induced oncogene cell transformation. In addition, melanoma, a deadly skin cancer, which arises from transformed melanocytes, is characterized by a resistance to chemotherapy. In our studies we have shown that NO can induce perturbation of melanocyte-extracellular matrix component interactions, which may contribute to loss of melanocytes or melanoma metastasis. Such NO effects appear to be modulated partly via cGMP. Moreover, we found that different guanylyl cyclase isoforms are responsible for cGMP synthesis in melanocytic cells. Normal human melanocytes and nonmetastatic melanoma cells predominantly express sGC, which appears to be associated with melanogenesis, whereas absence of NO-sensitive GC, but up-regulated activities of the natriuretic peptide-sensitive membrane guanylyl cyclase isoforms were found in highly metastatic phenotypes. Due to the growing interest in the regulation of signaling activities in normal and transformed cells under altered gravity conditions, we have further investigated whether the NO/cGMP signaling is involved in melanocyte response to gravitational stress. We found that normal human melanocytes and non-metastatic melanoma cell lines, but not highly metastatic cells

  14. Dysfunction of Autophagy: A Possible Mechanism Involved in the Pathogenesis of Vitiligo by Breaking the Redox Balance of Melanocytes.

    Science.gov (United States)

    Qiao, Zhuhui; Wang, Xiuxiu; Xiang, Leihong; Zhang, Chengfeng

    2016-01-01

    Vitiligo is a common chronic acquired pigmentation disorder characterized by loss of functional melanocytes from the epidermis and follicular reservoir. Among multiple hypotheses which have been proposed in the pathogenesis of vitiligo, autoimmunity and oxidative stress-mediated toxicity in melanocytes remain most widely accepted. Macroautophagy is a lysosome-dependent degradation pathway which widely exists in eukaryotic cells. Autophagy participates in the oxidative stress response in many cells, which plays a protective role in preventing damage caused by oxidative stress. Recent studies have enrolled autophagy as an important regulator in limiting damage caused by UV light and lipid oxidation, keeping oxidative stress in a steady state in epidermal keratinocytes and maintaining normal proliferation and aging of melanocytes. Impairment of autophagy might disrupt the antioxidant defense system which renders melanocytes to oxidative insults. These findings provide supportive evidence to explore new ideas of the pathogenesis of vitiligo and other pigmentation disorders.

  15. Suprabasal spread of melanocytes in dysplastic nevi and melanoma in situ: Ki-67-labeling rate of junctional melanocytes and suprabasal cells may be a helpful clue to the diagnosis.

    Science.gov (United States)

    Hall, Brian J; LeBoit, Philip E

    2014-08-01

    Multiple criteria on routinely stained sections allow one to make a diagnosis of a dysplastic or "Clark" nevus (CN) versus melanoma in situ (MIS), and one of these is suprabasal spread of melanocytes. The extent of suprabasal spread of melanocytes in otherwise conventional CN and the combination of a sensitive marker of melanocytes combined with Ki-67 to assess the differences between the proliferation of melanocytes at the junction and those above it have not yet been studied. Fifty classic examples of CN and 27 cases of MIS were culled from the files of a university-based dermatopathology practice. All cases were stained with a 2-color method (MART-1/tyrosinase red, Ki-67 brown) to evaluate morphologic and immunohistochemical differences in these lesions. Fifteen of 50 cases of benign CN demonstrated suprabasal spread compared with 27 of 27 cases of MIS. The majority of CNs with suprabasal spread (13 of 15) showed a 0% Ki-67-labeling rate among the suprabasal melanocytes, and the majority of MISs (23 of 27) showed a 20% or greater Ki-67-labeling rate in suprabasal cells. Suprabasal melanocytes can be seen by immunostaining in otherwise unremarkable CN, wherein they are not notable in routinely stained sections, but their proliferation rate is much less than in MIS.

  16. Follicle and melanocyte stem cells, and their application in neuroscience: A Web of Science-based literature analysis.

    Science.gov (United States)

    Wu, Weifu

    2012-12-05

    To identify global research trends of follicle and melanocyte stem cells, and their application in neuroscience. We performed a bibliometric analysis of studies from 2002 to 2011 on follicle and melanocyte stem cells, and their application in neuroscience, which were retrieved from the Web of Science, using the key words follicle stem cell or melanocyte stem cell, and neural, neuro or nerve. (a) peer-reviewed published articles on follicle and melanocyte stem cells, and their application in neuroscience, which were indexed in the Web of Science; (b) original research articles, reviews, meeting abstracts, proceedings papers, book chapters, editorial material, and news items. (a) articles that required manual searching or telephone access; (b) documents that were not published in the public domain; and (c) a number of corrected papers from the total number of articles. (1) Distribution of publications on follicle and melanocyte stem cells by years, journals, countries, institutions, institutions in China, and most cited papers. (2) Distribution of publications on the application of follicle and melanocyte stem cells in neuroscience by years, journals, countries, institutions, and most cited papers. Of the 348 publications from 2002 to 2011 on follicle and melanocyte stem cells, which were retrieved from the Web of Science, more than half were from American authors and institutes. The most prolific institutions in China for publication of papers on follicle and melanocyte stem cells were the Fourth Military Medical University and Third Military Medical University. The most prolific journals for publication of papers on follicle and melanocyte stem cells were the Journal of Investigative Dermatology, Pigment Cell & Melanoma Research. Of the 63 publications from 2002 to 2011 on the application of follicle and melanocyte stem cells in neuroscience, which were retrieved from the Web of Science, more than half were from American authors and institutes, and no papers were

  17. Follicle and melanocyte stem cells, and their application in neuroscience A Web of Science-based literature analysis

    Institute of Scientific and Technical Information of China (English)

    Weifu Wu

    2012-01-01

    OBJECTIVE: To identify global research trends of follicle and melanocyte stem cells, and their application in neuroscience. DATA RETRIEVAL: We performed a bibliometric analysis of studies from 2002 to 2011 on follicle and melanocyte stem cells, and their application in neuroscience, which were retrieved from the Web of Science, using the key words follicle stem cell or melanocyte stem cell, and neural, neuro or nerve. SELECTION CRITERIA: Inclusion criteria: (a) peer-reviewed published articles on follicle and melanocyte stem cells, and their application in neuroscience, which were indexed in the Web of Science; (b) original research articles, reviews, meeting abstracts, proceedings papers, book chapters, editorial material, and news items. Exclusion criteria: (a) articles that required manual searching or telephone access; (b) documents that were not published in the public domain; and (c) a number of corrected papers from the total number of articles. MAIN OUTCOME MEASURES: (1) Distribution of publications on follicle and melanocyte stem cells by years, journals, countries, institutions, institutions in China, and most cited papers. (2) Distribution of publications on the application of follicle and melanocyte stem cells in neuroscience by years, journals, countries, institutions, and most cited papers. RESULTS: Of the 348 publications from 2002 to 2011 on follicle and melanocyte stem cells, which were retrieved from the Web of Science, more than half were from American authors and institutes. The most prolific institutions in China for publication of papers on follicle and melanocyte stem cells were the Fourth Military Medical University and Third Military Medical University. The most prolific journals for publication of papers on follicle and melanocyte stem cells were the Journal of Investigative Dermatology, Pigment Cell & Melanoma Research. Of the 63 publications from 2002 to 2011 on the application of follicle and melanocyte stem cells in neuroscience, which

  18. 真皮黑素细胞的研究进展%Advances in dermal melanocytes

    Institute of Scientific and Technical Information of China (English)

    黄斐然; 王大光; 骆丹

    2015-01-01

    Dermal stem cells (DSCs) have great proliferation and multidirectional differentiation potential.Keratinocytes can induce DSCs to migrate and differentiate into melanocytes by expressing Wnt-3a and E-cadherins.Researches have indicated that the generation of dermal and epidermal melanocytes is dependent on different signaling pathways.More concretely,epidermal melanocyte generation is highly dependent on the stem cell factor (SCF) signaling pathway,while dermal melanocyte generation on the endothelin 3 (ET3) and hepatocyte growth factor (HGF) signaling pathways.The origin of nevus cells in pigmented nevus is not yet clear.Some studies suggest that nevus cells may derive from the stem cells which can differentiate into melanocytes.DSCs can differentiate into melanocytes,but further research is needed to determine whether they are the source of nevus cells.%真皮干细胞具有增殖和多向分化的潜能,角质形成细胞通过表达Wnt3a和E钙黏素来诱导真皮干细胞移行和分化为黑素细胞.研究表明,在黑素细胞形成过程中,真皮黑素细胞与表皮黑素细胞依赖的信号通路明显不同,表皮黑素细胞高度依赖干细胞因子信号通路,而真皮黑素细胞高度依赖内皮素3和肝细胞生长因子信号通路.目前关于色素痣痣细胞的来源尚不清楚,有研究认为,痣细胞来源于能分化为黑素细胞的干细胞,真皮干细胞能分化为黑素细胞,是否是痣细胞的来源尚需进一步研究.

  19. Co-conception d’itinéraires techniques de culture pure du niébé et du mucuna dans la zone cotonnière ouest du Burkina Faso : intérêts et limites.

    Directory of Open Access Journals (Sweden)

    Kalifa Coulibaly

    2012-12-01

    Full Text Available Résumé Dans un contexte d’accroissement de la pression humaine sur les espaces agricoles, de réduction des pâturages et de divagation des animaux, les paysans sont intéressés par des solutions qui leur permettent d’accroitre la production de biomasses à l’hectare tout en préservant la fertilité de leur sol. Dans la littérature, il est prouvé que les légumineuses jouent un rôle important dans l’amélioration des systèmes de culture. Mais, leur adoption est faible par les agriculteurs dans la zone cotonnière à l’ouest du Burkina Faso. L’objectif de cet article est de déterminer les performances agronomiques et économiques du niébé et du mucuna dans le cadre d’une démarche d’expérimentation chez et par les paysans (ECPP. Sur deux campagnes agricoles (2010 et 2011, nous avons utilisés les données sur les caractéristiques de 81 exploitations (45 pour le niébé et 36 pour le mucuna, les données économiques et agronomiques. Les résultats indiquent que le niébé est sensible à l’arrière effet de la précédente fertilisation contrairement au mucuna. Le mucuna qui aurait un réel intérêt pour les agriculteurs engagés dans l’intensification de leur élevage, offre l’opportunité de produire plus de biomasse (plus de 1,5 t/ha par unité de surface avec une meilleure qualité. Le niébé pourrait constituer une source de revenu pour les exploitations agricoles du Tuy avec une productivité de travail pouvant atteindre 10 379 FCFA par jour. Mais, dans un contexte de rareté de l’espace agricole, les expérimentations doivent se poursuivre pour tester les associations céréales/légumineuses avec les agriculteurs qui ont un rôle à jouer dans la création de ces cultures associées. Mots clés : Légumineuse, expérimentation chez et par les paysans, temps de travail, performance économique,

  20. Metabotropic glutamate receptor 6 signaling enhances TRPM1 calcium channel function and increases melanin content in human melanocytes.

    Science.gov (United States)

    Devi, Sulochana; Markandeya, Yogananda; Maddodi, Nityanand; Dhingra, Anuradha; Vardi, Noga; Balijepalli, Ravi C; Setaluri, Vijayasaradhi

    2013-05-01

    Mutations in TRPM1, a calcium channel expressed in retinal bipolar cells and epidermal melanocytes, cause complete congenital stationary night blindness with no discernible skin phenotype. In the retina, TRPM1 activity is negatively coupled to metabotropic glutamate receptor 6 (mGluR6) signaling through Gαo and TRPM1 mutations result in the loss of responsiveness of TRPM1 to mGluR6 signaling. Here, we show that human melanocytes express mGluR6, and treatment of melanocytes with L-AP4, a type III mGluR-selective agonist, enhances Ca(2+) uptake. Knockdown of TRPM1 or mGluR6 by shRNA abolished L-AP4-induced Ca(2+) influx and TRPM1 currents, showing that TRPM1 activity in melanocytes is positively coupled to mGluR6 signaling. Gαo protein is absent in melanocytes. However, forced expression of Gαo restored negative coupling of TRPM1 to mGluR6 signaling, but treatment with pertussis toxin, an inhibitor of Gi /Go proteins, did not affect basal or mGluR6-induced Ca(2+) uptake. Additionally, chronic stimulation of mGluR6 altered melanocyte morphology and increased melanin content. These data suggest differences in coupling of TRPM1 function to mGluR6 signaling explain different cellular responses to glutamate in the retina and the skin.

  1. Immunohistochemical expression of the glucose transporters Glut-1 and Glut-3 in human malignant melanomas and benign melanocytic lesions

    Directory of Open Access Journals (Sweden)

    Parente Paola

    2008-09-01

    Full Text Available Abstract Background Reported data indicate that cancer cells have increased rates of glucose metabolism, as determined by 18FDG-PET imaging in patients with malignancies. The results of many studies have demonstrated that the expression of glucose transporters, especially Glut-1, is increased in a variety of malignancies. This study was undertaken to assess the differential expression of Glut-1 and Glut-3 by benign and malignant melanocytic lesions. Methods Immunohistochemical staining for Glut-1 and Glut-3 was performed on paraffin-embedded tissue sections prepared from melanocytic nevi (12 cases, Spitz nevi (12 cases and primary cutaneous malignant melanomas (20 cases. Results We observed immunoreactivity for Glut-1 in all melanocytic nevi, 9 of the 12 Spitz nevi and in 9 of the 20 malignant melanomas, whereas Glut-3 was expressed in all the melanocytic lesions, both benign and malignant. Conclusion These findings indicate that the glucose transporters Glut-1 and Glut-3 play a role in the glucose metabolism of melanocytic cells. Glut-1 was present in the majority of benign nevi, whereas its expression was downregulated in 55% of malignant melanomas. Our results suggest that glucose transporter Glut-1 expression can significantly discriminate between human malignant melanoma and benign melanocytic nevi, and support the idea that additional mechanisms other than Glut-1 may contribute to glucose uptake in melanomas.

  2. Nests with numerous SOX10 and MiTF-positive cells in lichenoid inflammation: pseudomelanocytic nests or authentic melanocytic proliferation?

    Science.gov (United States)

    Silva, Claudine Yap; Goldberg, Lynne J; Mahalingam, Meera; Bhawan, Jag; Wolpowitz, Deon

    2011-10-01

    Pseudomelanocytic nests in the setting of lichenoid inflammation can mimic atypical melanocytic proliferations. Both melanocytic and cytokeratin immunohistochemical stains may be utilized to differentiate these entities. Unlike true melanocytic nests, pseudomelanocytic nests contain Melanoma Antigen Recognized by T-cells 1 (MART-1)/ Melan-A-positive cells and cells positive for pan-cytokeratins, CD3 and/or CD68. Recently, rare (1-2 cells/nest) microphthalmia- associated transcription factor (MiTF)-positive cells were also reported in pseudomelanocytic nests. We present a 48-year-old man with a 2 × 3 cm violaceous to hyperpigmented, non-blanching, polygonal patch on the neck. Histopathology showed focal epidermal atrophy, irregularly distributed junctional nests and a lichenoid infiltrate with colloid bodies. Immunoperoxidase studies revealed occasional pan-cytokeratin and MART-1/Melan-A-positive staining in nests as well as focal S-100 protein-positive cells. Importantly, the majority of nests showed numerous cells positive for MiTF and SOX10 (>2 cells/nest and some the majority of cells). This combined staining pattern confounds the above-described immunohistochemical distinction between pseudo and true melanocytic nests. Clinically felt to represent unilateral lichen planus pigmentosus/erythema dyschromicum perstans and not malignant melanoma in situ, this lesion highlights the importance of clinicopathologic correlation and suggests either a new melanocytic entity or a novel pattern of benign melanocytic reorganization in a subset of lichenoid dermatitides.

  3. Universality in pure gravity mediation

    Energy Technology Data Exchange (ETDEWEB)

    Evans, Jason L.; Olive, Keith A. [University of Minnesota, William I. Fine Theoretical Physics Institute, School of Physics and Astronomy, Minneapolis, MN (United States); Ibe, Masahiro [University of Tokyo, ICRR, Kashiwa (Japan); University of Tokyo, Kavli IPMU, TODIAS, Kashiwa (Japan); Yanagida, Tsutomu T. [University of Tokyo, Kavli IPMU, TODIAS, Kashiwa (Japan)

    2013-07-15

    If low-energy supersymmetry is realized in nature, the apparent discovery of a Higgs boson with mass around 125 GeV suggests a supersymmetric mass spectrum in the TeV or multi-TeV range. Multi-TeV scalar masses are a necessary component of supersymmetric models with pure gravity mediation or in any model with strong moduli stabilization. Here, we show that full scalar mass universality remains viable as long as the ratio of Higgs vevs, tan{beta}, is relatively small (

  4. Toxicological evaluation of pure hydroxytyrosol.

    Science.gov (United States)

    Auñon-Calles, David; Canut, Lourdes; Visioli, Francesco

    2013-05-01

    Of all the phenolic constituents of olives and extra virgin olive oil, hydroxytyrosol is currently being actively exploited as a potential supplement or preservative to be employed in the nutraceutical, cosmeceutical, and food industry. In terms of safety profile, hydroxytyrosol has only been investigated as the predominant part of raw olive mill waste water extracts, due to the previous unavailability of appropriate quantities of the pure compound. We report the toxicological evaluation of hydroxytyrosol and, based on the results, propose a No Observed Adverse Effects Level (NOAEL) of 500mg/kg/d.

  5. Synthesis of Enantiomerically Pure Anthracyclinones

    Science.gov (United States)

    Achmatowicz, Osman; Szechner, Barbara

    The anthracycline antibiotics are among the most important clinical drugs used in the treatment of human cancer. The search for new agents with improved therapeutic efficacy and reduced cardiotoxicity stimulated considerable efforts in the synthesis of new analogues. Since the biological activity of anthracyclines depends on their natural absolute configuration, various strategies for the synthesis of enantiomerically pure anthracyclinones (aglycones) have been developed. They comprise: resolution of racemic intermediate, incorporation of a chiral fragment derived from natural and non-natural chiral pools, asymmetric synthesis with the use of a chiral auxiliary or a chiral reagent, and enantioselective catalysis. Synthetic advances towards enantiopure anthracyclinones reported over the last 17 years are reviewed.

  6. Pure dysarthria due to an insular infarction.

    Science.gov (United States)

    Hiraga, Akiyuki; Tanaka, Saiko; Kamitsukasa, Ikuo

    2010-06-01

    Cortical infarction presenting with pure dysarthria is rarely reported. Previous studies have reported pure dysarthria due to cortical stroke at the precentral gyrus or middle frontal gyrus. We report a 72-year-old man who developed pure dysarthria caused by an acute cortical infarction in the insular cortex. The role of the insula in language has been difficult to assess clinically because of the rarity of pure insular strokes. Our patient showed pure dysarthria without aphasia, indicating that pure dysarthria can be the sole manifestation of insular infarctions.

  7. Primary melanocytic tumors of the central nervous system: A neuroradiological and clinicopathological study of five cases and brief review of literature

    Directory of Open Access Journals (Sweden)

    Sushila Jaiswal

    2011-01-01

    Full Text Available Primary melanocytic tumors of the central nervous system (CNS are uncommon lesions. These lesions arise from the melanocytes located within leptomeninges and include diffuse melanocytosis and meningeal melanomatosis (seen in neurocutaneous melanosis, melanocytoma, and malignant melanoma. To study, the clinical course, neuroradiological features, morphology and immunohistochemistry of primary melanocytic tumor of CNS. Demographic, clinical and surgico-pathologic findings of five patients with melanocytic tumors seen between 1996 and 2003 were studied. In this study, five cases of primary melanocytic tumors have been reported: four cases of malignant melanoma and one case of melanocytoma. Three of the 5 cases were intracranial and 2 were spinal. The mean age in the present study was 26 years. Presenting features varied according to the location. Primary melanocytic tumor of CNS are rare. Whenever possible, complete surgical excision is the best treatment.

  8. "Sky Full of Stars" Pattern: Dermoscopic Findings in a Desmoplastic Giant Congenital Melanocytic Nevus.

    Science.gov (United States)

    Martín-Carrasco, Pablo; Bernabeu-Wittel, José; Dominguez-Cruz, Javier; Zulueta Dorado, Teresa; Conejo-Mir Sanchez, Julian

    2017-02-27

    Desmoplastic giant congenital melanocytic nevus (DGCN) is an uncommon variant of congenital nevus, presenting as a progressive induration and hypopigmentation of the lesion that occasionally causes hair loss and even total or partial disappearance of the nevus. A 6-month-old girl with a giant congenital melanocytic nevus that involved the entire posterior side of the right thigh was seen in our department. Nine months later, the peripheral area of the nevus presented as a marked induration with hypopigmentation. Dermoscopy demonstrated a reticular pattern exclusively located in the perifollicular areas, with a radial distribution from the follicular ostium that mimicked a "sky full of stars." We report a case of DGCN, including a dermoscopic description of the findings noted in the indurated and hypopigmented areas that appear as a "sky full of stars" image.

  9. Characterization of Streptococcus thermophilus two-component systems: In silico analysis, functional analysis and expression of response regulator genes in pure or mixed culture with its yogurt partner, Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Thevenard, Benoît; Rasoava, Niriaina; Fourcassié, Pascal; Monnet, Véronique; Boyaval, Patrick; Rul, Françoise

    2011-12-02

    The lactic acid bacterium Streptococcus thermophilus (S. thermophilus) is widely used in the dairy industry. As a food bacterium, it has to cope with changing environments such as milk, yogurt, as well as the digestive tract, after the product has been ingested. In bacteria, two-component systems (TCS) are one of the most prevalent mechanisms to sense and respond appropriately to a wide range of signals. They are typically composed of a sensor kinase (HK) that detects a stimulus and a response regulator (RR) which acts as a transcriptional regulator. Our objective was to make an inventory of the TCS present in S. thermophilus LMD-9 and investigate the contribution of each TCS to LMD-9 growth in milk. For that purpose, we performed in silico, transcriptomic as well as functional analysis. The LMD-9 genome presented 6 complete TCS with both HK and RR (TCS 2, 4, 5, 6, 7, and 9) and 2 orphan RRs (RR01 and 08) with truncated HK. Our in silico analysis revealed that for 5 TCS out of the 8, orthologs with known functions were found in other bacterial species whereas for TCS02, 4 and 6 the function of the orthologs are unidentified. Transcriptomic studies (using quantitative PCR) revealed that all S. thermophilus LMD-9 response regulator genes were expressed in milk; they were expressed at different levels and with different profiles during growth. In mixed culture with Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus), the S. thermophilus partner in yogurt, the expression of four S. thermophilus LMD-9 response regulator increased; two of them, rr02 and rr09, increased by a factor of 6. These results indicate that the presence of L. bulgaricus induces regulatory changes in S. thermophilus. We also demonstrated that a response regulator (rr02) can exert its regulatory function on its target genes even when expressed at very low levels. We showed that RR05-an ortholog of Bacillus subtilis YycF or Staphylococcus aureus WalR-was essential for the growth of S

  10. Multiple giant congenital melanocytic nevi with central nervous system melanosis: A case report

    Directory of Open Access Journals (Sweden)

    Ahuja S

    2003-10-01

    Full Text Available A case of multiple giant congenital melanocytic naevi in whom central nervous system melanosis was detected at 6 weeks of age is described. The infant was asymptomatic, but presence of risk factors such as multiple naevi, giant naevi and naevi on scalp and posterior axial location prompted a magnetic resonance imaging study of the brain. To our knowledge, neurocutaneous melanosis at such a young age has not been reported in Indian literature.

  11. NFIB is a governor of epithelial–melanocyte stem cell behaviour in a shared niche

    OpenAIRE

    Chang, Chiung-Ying; Pasolli, H. Amalia; Giannopoulou, Eugenia G.; Guasch, Géraldine; Gronostajski, Richard M; Elemento, Olivier; Fuchs, Elaine

    2013-01-01

    Adult stem cells reside in specialized niches where they receive environmental cues to maintain tissue homeostasis. In mammals, the stem cell niche within hair follicles is home to epithelial hair follicle stem cells and melanocyte stem cells, which sustain cyclical bouts of hair regeneration and pigmentation1–4. To generate pigmented hairs, synchrony is achieved such that upon initiation of a new hair cycle, stem cells of each type activate lineage commitment2,5. Dissecting the inter-stem-ce...

  12. Value of dopachrome tautomerase detection in the assessment of melanocytic tumors.

    Science.gov (United States)

    Filimon, Anca; Zurac, Sabina A; Milac, Adina L; Sima, Livia E; Petrescu, Stefana M; Negroiu, Gabriela

    2014-06-01

    Dopachrome tautomerase (DCT) and tyrosinase (Tyr) are melanogenic enzymes and structurally related melanosomal proteins. The present study investigates DCT expression comparatively with Tyr, the most tested melanoma biomarker, aiming to evaluate DCT potential in the assessment of melanocytic tumors and gain insights into the molecular and pathological characterization of DCT-phenotype in tumor progression. DCT and Tyr are simultaneously analyzed in melanoma cell lines by semiquantitative RT-PCR, western blot, and N-glycan analysis, and in cell populations of melanocytic tumors by immunohistofluorescence using a novel anti-hDCT antibody against an extended sequence within DCT luminal domain. DCT, unlike Tyr, is fully processed along the secretory pathway in both pigmented and amelanotic melanoma cells. In 53 nevi and 116 primary malignant melanomas, 81% and 52%, respectively, are DCT+/Tyr+, showing that DCT is a stable antigen, retained by most tumors and partially expressed in Tyr-negative cell populations. The DCT/Tyr disjunction is a process correlated with melanocyte neoplastic transformation and malignant progression. A tumor architecture--DCT-phenotype-containing DCT+/Tyr- cell populations selected into the innermost dermis from double-positive cells is detected in 35% of DCT+/Tyr+ specimens. The DCT-phenotype is associated with enhanced neurotization in benign nevi and with ulceration in thin malignant melanomas. The intradermal DCT+/Tyr- clones in superficial melanomas acquire the expression and specific subcellular distribution of unfavorable prognostic markers. DCT assessment shows specific antigen patterns with potential significance in the outcome of melanocytic lesions, connecting DCT, a mediator of a melanoma stress-resistant pathway, and an antiapoptotic molecule to DCT- phenotypes that are possibly more stable and stress resistant.

  13. Prognostic implications of cytopathologic classification of melanocytic uveal tumors evaluated by fine-needle aspiration biopsy

    OpenAIRE

    2013-01-01

    PURPOSE: Determine whether cytopathologic classification of melanocytic uveal tumors evaluated by fine-needle aspiration biopsy (FNAB) is a significant prognostic factor for death from metastasis. METHODS: Retrospective analysis of cases of clinically diagnosed uveal melanoma evaluated by fine-needle aspiration biopsy from 1980 to 2006. Main outcome evaluated was death from metastasis. Associations between baseline clinical variables and cytopathologic classification were evaluated using cros...

  14. Comparison of visual effects of immersion fluids for dermoscopic examination of acral volar melanocytic lesions

    Directory of Open Access Journals (Sweden)

    Tzu-Hsiu Chen

    2014-06-01

    Conclusion: The use of either mineral oil or ultrasound jelly as interface provides acceptable visual effects for the dermoscopic examination of acral volar melanocytic lesions. The use of the polarized light mode reduced the reflection and scattering of light, resulting in better visual effect than that achieved using the nonpolarized light mode. In the early diagnosis of acral melanoma, choosing the appropriate application of immersion fluid and observation mode yields the optimal visual effect.

  15. Detection of serum anti-melanocyte antibodies and identification of related antigens in patients with vitiligo.

    Science.gov (United States)

    Zhu, M C; Liu, C G; Wang, D X; Zhan, Z

    2015-12-07

    We detected autoantibodies against melanocytes in serum samples obtained from 50 patients, including 4 with HBV, with vitiligo and identified the associated membrane antigens. Heat shock protein 70 (HSP70) and anti-tyrosinase-related protein 1 (TRP-1) antibody levels were analyzed. The associated antigens in normal human melanocyte were identified by immunofluorescence. Autoantibodies against melanocyte membrane and cytoplasmic proteins were detected by western blot. Membrane antigens with higher frequencies were identified by protein mass spectrometry. The HSP70 and anti-TRP-1 antibody levels (N = 70; 10 with HBV) were detected by ELISA. The specific antigens were detected in melanocyte cytoplasm and membrane (40/50; 80% incidence; western blot). The autoantibodies reacted with several membrane antigens with approximate molecular weights (Mr) of 86,000, 75,000, 60,000, 52,000, and 44,000 (strip positive rates: 36, 58, 22, 2, and 2%, respectively). Thirty percent of the patients showed the presence of cytoplasmic antigens (Mr: 110,000, 90,000, 75,000, 50,000, and 400,000; strip positive rates: 12, 4, 12, 10, and 2%, respectively). Fifteen and 5% of the healthy subjects showed positive expression of membrane and cytoplasmic antigens, respectively. Protein mass spectrometry predicted membrane proteins with Mr of 86,000 and 75,000 and 60,000 to be Lamin A /C and Vimentin X1, respective. High titers of anti-TRP-1 antibody were detected and showed positive correlation with HSP70 (r = 0. 927, P vitiligo, which might assist future investigations into autoimmune pathogenesis of vitiligo and formation of autoantibodies. HBV infection was correlated to vitiligo.

  16. Toxicological and melanin synthesis effects of Polygonum multiflorum root extracts on zebrafish embryos and human melanocytes

    Directory of Open Access Journals (Sweden)

    Thanh Thi Hoai Dang

    2016-09-01

    Full Text Available Polygonum multiflorum (PM has been commmonly used as folk medicine for treatment of various conditions, such as early graying of hair in humans. However, there have been limited studies which have evaluated the toxicological and biological effects of PM in vitro as well as in vivo. In this study, PM root extracts in ethyl acetate (PM-E and in distilled water (PM-W were examined for their effects on the development of teratogenic defects/deaths. Additionally, they were evaluated for their effects on melanin formation in human melanocytes and pigmentation in embryos/larvae of wild type strain AB zebrafish (Danio rerio. Our results showed that PM root extracts at concentrations of 40 mg/L and 105 mg/L induced the development of teratogenic defects, including yolk sac edema (or heart edema, hemovascular defects, necrosis and abnormal trunk in zebrafish embryos at 4 days post fertilization; teratogenic indexes (TIs were 1.43 and 0.63 for ethyl acetate extract and distilled water extract, respectively. Our results also demonstrated that PM-W significantly increased the pigmentation level of embryos/larvae and induced melanin formation in human melanocytes. The amount of melanin in PM-W-exposed embryos/larvae was 2.2-fold and 1.71-fold greater than those in the control embryos/larvae and control melanocytes, respectively. Our study also showed that the increased level of pigmentation in PM-W embryos/larvae or melanin biosynthesis in melanocytes were both regulated by activation of tyrosinase. Conclusively, our study suggests that PM root extracts could be used as potential agents for treatment of early hair graying as well as various other diseases related to loss of pigmentation. However, these PM root extracts may also have some negative effects on embryos; therefore it should be careful when using for women during pregnancy. [Biomed Res Ther 2016; 3(9.000: 808-818

  17. Uncovering the Role of BMP Signaling in Melanocyte Development and Melanoma Tumorigenesis

    Science.gov (United States)

    2015-06-01

    Please see appended curriculum vitae for Dr. Ceol. Please see appended meeting abstracts. 12 CRAIG JOSEPH CEOL Assistant Professor Program in... haired , fair-skinned individuals. Our data support the notion that disrupted cAMP signaling is a more pervasive contributor to melanoma, including in...Biology, University of Massachusetts Medical School, Worcester, MA, USA Melanocytes, which can be lost during hair graying, injury and disease-related

  18. Discovering novel phenotypes with automatically inferred dynamic models: a partial melanocyte conversion in Xenopus

    Science.gov (United States)

    Lobo, Daniel; Lobikin, Maria; Levin, Michael

    2017-01-01

    Progress in regenerative medicine requires reverse-engineering cellular control networks to infer perturbations with desired systems-level outcomes. Such dynamic models allow phenotypic predictions for novel perturbations to be rapidly assessed in silico. Here, we analyzed a Xenopus model of conversion of melanocytes to a metastatic-like phenotype only previously observed in an all-or-none manner. Prior in vivo genetic and pharmacological experiments showed that individual animals either fully convert or remain normal, at some characteristic frequency after a given perturbation. We developed a Machine Learning method which inferred a model explaining this complex, stochastic all-or-none dataset. We then used this model to ask how a new phenotype could be generated: animals in which only some of the melanocytes converted. Systematically performing in silico perturbations, the model predicted that a combination of altanserin (5HTR2 inhibitor), reserpine (VMAT inhibitor), and VP16-XlCreb1 (constitutively active CREB) would break the all-or-none concordance. Remarkably, applying the predicted combination of three reagents in vivo revealed precisely the expected novel outcome, resulting in partial conversion of melanocytes within individuals. This work demonstrates the capability of automated analysis of dynamic models of signaling networks to discover novel phenotypes and predictively identify specific manipulations that can reach them. PMID:28128301

  19. Characterization and expression of soluble guanylate cyclase in skins and melanocytes of sheep.

    Science.gov (United States)

    Yang, Shanshan; Zhang, Junzhen; Ji, Kaiyuan; Jiao, Dingxing; Fan, Ruiwen; Dong, Changsheng

    2016-04-01

    The study reported the characterization of soluble guanylate cyclase (sGC) with the size of CDS of 1860bp, encoding a protein of 620 amino acids and containing several conserved functional domains including HNOB, HNOBA, and CHD. Quantitative real time PCR analysis of sGC showed that the expression of sGC mRNA is higher (∼5 fold) in white sheep skin relative to black sheep skin with significant difference (Pmelanocytes in vitro of sheep skin. Over expression of sGC in melanocytes resulted in decreased expression of key melanogenic genes including microphthalmia transcription factor (MITF), tyrosinase (TYR), tyrosinase related protein 1(TYRTP1), and tyrosinase related protein 2(TYRP2) both at mRNA and protein level. Moreover, the melanocytes was capable of producing cGMP and cAMP. The observed differential expression and localization of sGC in sheep skins and melanocytes and the capability of producing cGMP and cAMP, which suggested a potential role for this gene in hair color regulation.

  20. Effect of thioridazine on antioxidant status of HEMn-DP melanocytes.

    Science.gov (United States)

    Otręba, Michał; Beberok, Artur; Wrześniok, Dorota; Rok, Jakub; Buszman, Ewa

    2015-10-01

    Thioridazine as an antipsychotic agent was extensively used to treat various psychotic disorders, e.g. schizophrenia. However, the therapy with this drug can induce serious side effects such as extrapyramidal symptoms or ocular and skin disorders, which mechanisms are still not fully established. To gain inside the molecular mechanisms underlying thioridazine toxicity, we examined the effect of this drug on cell viability, antioxidant defence system as well as melanogenesis in normal human melanocytes. It was demonstrated that thioridazine induces concentration-dependent loss in cell viability. The value of EC50 was calculated to be 2.24 μM. To study the effect of thioridazine on antioxidant defence system in melanocytes, the level of hydrogen peroxide and the activities of antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase were determined. The drug in concentrations of 0.1, 0.25, 1.0 and 2.5 μM caused changes in cellular antioxidant defence system indicating the induction of oxidative stress. It was also shown that the analysed neuroleptic in concentrations of 1.0 and 2.5 μM significantly inhibited melanogenesis. The observed changes in cell viability, antioxidant defence system and melanization in normal human melanocytes after thioridazine treatment may explain an important role of reactive oxygen species as well as melanin in mechanisms involved in this drug side effects directed on pigmented tissues.

  1. Mechanical properties of growing melanocytic nevi and the progression to melanoma

    Science.gov (United States)

    Taloni, Alessandro; Alemi, Alexander; Ciusani, Emilio; Sethna, James P.; Zapperi, Stefano; La Porta, Caterina A. M.; National Research Council Of Italy Team; Lassp, Department Of Physics, Cornell University Team; Istituto Neurologico Carlo Besta Collaboration; Department Of Biosciences, University Of Milano Team

    2015-03-01

    Melanocytic nevi are benign proliferations that sometimes turn into malignant melanoma in a way that is still unclear from the biochemical and genetic point of view. Diagnostic and prognostic tools are then mostly based on dermoscopic examination and morphological analysis of histological tissues. To investigate the role of mechanics and geometry in the morpholgical dynamics of melanocytic nevi, we present a computational model for cell proliferation in a layered non-linear elastic tissue. Our simulations show that the morphology of the nevus is correlated to the initial location of the proliferating cell starting the growth process and to the mechanical properties of the tissue. We also demonstrate that melanocytes are subject to compressive stresses that fluctuate widely in the nevus and depend on the growth stage. Numerical simulations of cells in the epidermis releasing matrix metalloproteinases display an accelerated invasion of the dermis by destroying the basal membrane. Moreover, we show experimentally that osmotic stress and collagen inhibit growth in primary melanoma cells while the effect is much weaker in metastatic cells.

  2. NMR metabolic fingerprints of murine melanocyte and melanoma cell lines: application to biomarker discovery

    Science.gov (United States)

    Santana-Filho, Arquimedes Paixão de; Jacomasso, Thiago; Riter, Daniel Suss; Barison, Andersson; Iacomini, Marcello; Winnischofer, Sheila Maria Brochado; Sassaki, Guilherme Lanzi

    2017-01-01

    Melanoma is the most aggressive type of skin cancer and efforts to improve the diagnosis of this neoplasia are largely based on the use of cell lines. Metabolomics is currently undergoing great advancements towards its use to screening for disease biomarkers. Although NMR metabolomics includes both 1D and 2D methodologies, there is a lack of data in the literature regarding heteronuclear 2D NMR assignments of the metabolome from eukaryotic cell lines. The present study applied NMR-based metabolomics strategies to characterize aqueous and lipid extracts from murine melanocytes and melanoma cell lines with distinct tumorigenic potential, successfully obtaining fingerprints of the metabolites from the extracts of the cell lines by means of 2D NMR HSQC correlation maps. Relative amounts of the identified metabolites were compared between the 4 cell lines. Multivariate analysis of 1H NMR data was able not only to differentiate the melanocyte cell line from the tumorigenic ones but also distinguish among the 3 tumorigenic cell lines. We also investigated the effects of mitogenic agents, and found that they can markedly influence the metabolome of the melanocyte cell line, resembling the pattern of most proliferative cell lines. PMID:28198377

  3. Discovering novel phenotypes with automatically inferred dynamic models: a partial melanocyte conversion in Xenopus

    Science.gov (United States)

    Lobo, Daniel; Lobikin, Maria; Levin, Michael

    2017-01-01

    Progress in regenerative medicine requires reverse-engineering cellular control networks to infer perturbations with desired systems-level outcomes. Such dynamic models allow phenotypic predictions for novel perturbations to be rapidly assessed in silico. Here, we analyzed a Xenopus model of conversion of melanocytes to a metastatic-like phenotype only previously observed in an all-or-none manner. Prior in vivo genetic and pharmacological experiments showed that individual animals either fully convert or remain normal, at some characteristic frequency after a given perturbation. We developed a Machine Learning method which inferred a model explaining this complex, stochastic all-or-none dataset. We then used this model to ask how a new phenotype could be generated: animals in which only some of the melanocytes converted. Systematically performing in silico perturbations, the model predicted that a combination of altanserin (5HTR2 inhibitor), reserpine (VMAT inhibitor), and VP16-XlCreb1 (constitutively active CREB) would break the all-or-none concordance. Remarkably, applying the predicted combination of three reagents in vivo revealed precisely the expected novel outcome, resulting in partial conversion of melanocytes within individuals. This work demonstrates the capability of automated analysis of dynamic models of signaling networks to discover novel phenotypes and predictively identify specific manipulations that can reach them.

  4. ABSOLUTELY E-PURE MODULES AND E-PURE SPLIT MODULES

    Institute of Scientific and Technical Information of China (English)

    Yan Hangyu

    2011-01-01

    We first introduce the concepts of absolutely E-pure modules and Epure split modules. Then, we characterize the IF rings in terms of absolutely E-pure modules. The E-pure split modules are also characterized.

  5. Pure optical dynamical color encryption

    Science.gov (United States)

    Mosso, Fabian; Tebaldi, Myrian; Fredy Barrera, John; Bolognini, Néstor; Torroba, Roberto

    2011-07-01

    We introduce a way to encrypt-decrypt a color dynamical phenomenon using a pure optical alternative. We split the three basic chromatic channels composing the input, and then each channel is processed through a 4f encoding method and a theta modulation applied to the each encrypted frame in every channel. All frames for a single channel are multiplexed. The same phase mask is used to encode all the information. Unlike the usual procedure we do not multiplex the three chromatic channels into a single encoding media, because we want to decrypt the information in real time. Then, we send to the decoding station the phase mask and the three packages each one containing the multiplexing of a single channel. The end user synchronizes and decodes the information contained in the separate channels. Finally, the decoding information is conveyed together to bring the decoded dynamical color phenomenon in real-time. We present material that supports our concepts.

  6. 76 FR 69284 - Pure Magnesium From China

    Science.gov (United States)

    2011-11-08

    ... COMMISSION Pure Magnesium From China Determination On the basis of the record \\1\\ developed in the subject... order on pure magnesium from China would be likely to lead to continuation or recurrence of material... USITC Publication 4274 (October 2011), entitled Pure Magnesium from China: Investigation No....

  7. MC1R and the response of melanocytes to ultraviolet radiation

    Energy Technology Data Exchange (ETDEWEB)

    Rouzaud, Francois [Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Building 37, Room 2132, Bethesda, MD 20892 (United States); Kadekaro, Ana Luisa [Department of Dermatology, University of Cincinnati, College of Medicine, Cincinnati, OH 45267 (United States); Abdel-Malek, Zalfa A. [Department of Dermatology, University of Cincinnati, College of Medicine, Cincinnati, OH 45267 (United States); Hearing, Vincent J. [Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Building 37, Room 2132, Bethesda, MD 20892 (United States)]. E-mail: hearingv@nih.gov

    2005-04-01

    The constitutive color of our skin plays a dramatic role in our photoprotection from solar ultraviolet radiation (UVR) that reaches the Earth and in minimizing DNA damage that gives rise to skin cancer. More than 120 genes have been identified and shown to regulate pigmentation, one of the key genes being melanocortin 1 receptor (MC1R) that encodes the melanocortin 1 receptor (MC1R), a seven-transmembrane G protein-coupled receptor expressed on the surface of melanocytes. Modulation of MC1R function regulates melanin synthesis by melanocytes qualitatively and quantitatively. The MC1R is regulated by the physiological agonists {alpha}-melanocyte-stimulating hormone ({alpha}MSH) and adrenocorticotropic hormone (ACTH), and antagonist agouti signaling protein (ASP). Activation of the MC1R by binding of an agonist stimulates the synthesis of eumelanin primarily via activation of adenylate cyclase. The significance of cutaneous pigmentation lies in the photoprotective effect of melanin, particularly eumelanin, against sun-induced carcinogenesis. Epidermal melanocytes and keratinocytes respond to UVR by increasing their expression of {alpha}MSH and ACTH, which up-regulate the expression of MC1R, and consequently enhance the response of melanocytes to melanocortins. Constitutive skin pigmentation dramatically affects the incidence of skin cancer. The pigmentary phenotype characterized by red hair, fair complexion, inability to tan and tendency to freckle is an independent risk factor for all skin cancers, including melanoma. The MC1R gene is highly polymorphic in human populations, and allelic variation at this locus accounts, to a large extent, for the variation in pigmentary phenotypes and skin phototypes (SPT) in humans. Several allelic variants of the MC1R gene are associated with the red hair and fair skin (RHC) phenotype, and carrying one of these variants is thought to diminish the ability of the epidermis to respond to DNA damage elicited by UVR. The MC1R gene is

  8. Melanocyte transformation requires complete loss of all pocket protein function via a mechanism that mitigates the need for MAPK pathway activation.

    Science.gov (United States)

    Tonks, I D; Mukhopadhyay, P; Schroder, W A; Sorolla, A; Mould, A W; Handoko, H Y; Ferguson, B; Muller, H K; Keith, P; Hayward, N K; Walker, G J; Kay, G F

    2017-06-29

    Deregulation of p16INK4A is a critical event in melanoma susceptibility and progression. It is generally assumed that the major effect of loss of p16 function is mediated through the CDK-cyclin pathway via its influence on the pocket protein (PP) pRb. However, there are also two other PPs, p107 and p130, which, when phosphorylated by CDK-cyclin complexes, play a role in permitting cell progression. Cohorts of mice carrying melanocyte-specific knockouts (KOs) of various combinations of the three PPs were generated. Mice null for pRb, p107, p130 or any combination of double mutants did not develop melanoma. Surprisingly, melanocyte-specific loss of all three PPs facilitated melanoma development (median age of onset 308 days, penetrance 40% at 1 year). Tumorigenesis was exacerbated by Trp53 co-deletion (median age of onset 275 days, penetrance 82% at 1 year), with cell culture studies indicating that this difference may result from the apoptotic role of Trp53. Melanomas in PP;Trp53-deficient mice lacked either Ras or Braf mutations, and hence developed in the absence of constitutive MAPK pathway activation. The lag period between induction of total PP or PP/Trp53 KO and melanoma development indicates that additional genetic or epigenetic alterations may account for neoplastic progression. However, exome sequencing of PP;Trp53 KO melanomas failed to reveal any additional recurrent driver mutations. Analysis of the putative mutation signature of the PP;Trp53 KO melanomas suggests that melanocytes are primed for transformation via a mutagenic mechanism involving an excess of T>G substitutions, but not involving a preponderance of C>T substitutions at CpG sites, which is the case for most spontaneous cancers not driven by a specific carcinogen. In sum, deregulation of all three PPs appears central to neoplastic progression for melanoma, and the customary reference to the p16(INKA)/CDK4/pRB pathway may no longer be accurate; all PPs are potentially critical targets of CDK

  9. Interleukin-18 augments growth ability of primary human melanocytes by PTEN inactivation through the AKT/NF-κB pathway.

    Science.gov (United States)

    Zhou, Jia; Shang, Jing; Song, Jing; Ping, Fengfeng

    2013-02-01

    Normal human skin relies on melanocytes to provide photoprotection and thermoregulation by producing melanin. The growth and behavior of melanocytes are controlled by many factors. Interleukin-18 (IL-18) is expressed in both immune and non-immune cells and participates in the adjustment of multitude cellular functions. Nonetheless, the regulative roles of IL-18 in melanogenesis and growth of melanocytes have not been explored. The present study was conducted to investigate the effects of IL-18 on melanocytes and elucidate the underlying mechanisms. We proved that IL-18 increased the tyrosinase activity and melanin content in normal human foreskin-derived epidermal melanocytes (NHEM). Treatment with IL-18 (20 ng/ml) enhanced the expression of c-Kit, microphtalmia-associated transcription factor (MITF) and its downstream tyrosinase-related protein 1 (TRP-1), and TRP-2. In addition, IL-18 induced NHEM migration at concentration of 20 ng/ml. These results indicated a promotive action of IL-18 on melanogenesis in NHEM. Our data revealed that IL-18 stimulated ERK1/2 and NF-κB activation, improved p-Akt, p70 S6K and anti-apoptotic Bcl-2 levels, and deactivated phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in NHEM. Besides, IL-18 increased level of PTEN phosphorylation to protect NHEM from damage induced by H(2)O(2). These results in vitro showed the accommodation of IL-18 in melanocytes growth. Therefore, we suggested an important regulating action of IL-18 to melanogenesis and cell growth ability of skin melanocytes.

  10. Dual Target Search is Neither Purely Simultaneous nor Purely Successive.

    Science.gov (United States)

    Cave, Kyle R; Menneer, Tamaryn; Nomani, Mohammad S; Stroud, Michael J; Donnelly, Nick

    2017-08-31

    Previous research shows that visual search for two different targets is less efficient than search for a single target. Stroud, Menneer, Cave and Donnelly (2012) concluded that two target colours are represented separately based on modeling the fixation patterns. Although those analyses provide evidence for two separate target representations, they do not show whether participants search simultaneously for both targets, or first search for one target and then the other. Some studies suggest that multiple target representations are simultaneously active, while others indicate that search can be voluntarily simultaneous, or switching, or a mixture of both. Stroud et al.'s participants were not explicitly instructed to use any particular strategy. These data were revisited to determine which strategy was employed. Each fixated item was categorised according to whether its colour was more similar to one target or the other. Once an item similar to one target is fixated, the next fixated item is more likely to be similar to that target than the other, showing that at a given moment during search, one target is generally favoured. However, the search for one target is not completed before search for the other begins. Instead, there are often short runs of one or two fixations to distractors similar to one target, with each run followed by a switch to the other target. Thus, the results suggest that one target is more highly weighted than the other at any given time, but not to the extent that search is purely successive.

  11. Decryption of pure-position permutation algorithms

    Institute of Scientific and Technical Information of China (English)

    赵晓宇; 陈刚; 张亶; 王肖虹; 董光昌

    2004-01-01

    Pure position permutation image encryption algorithms, commonly used as image encryption investigated in this work are unfortunately frail under known-text attack. In view of the weakness of pure position permutation algorithm,we put forward an effective decryption algorithm for all pure-position permutation algorithms. First, a summary of the pure position permutation image encryption algorithms is given by introducing the concept of ergodic matrices. Then, by using probability theory and algebraic principles, the decryption probability of pure-position permutation algorithms is verified theoretically; and then, by defining the operation system of fuzzy ergodic matrices, we improve a specific decryption al-gorithm. Finally, some simulation results are shown.

  12. Effects of broadband ultraviolet B on non-canonical Wnt pathways in human epidermal melanocytes%宽谱中波紫外线对人表皮黑素细胞非经典Wnt通路的作用研究

    Institute of Scientific and Technical Information of China (English)

    王辉; 林彤; 王千秋

    2015-01-01

    目的 探讨宽谱中波紫外线(BB-UVB)照射对黑素细胞增殖率、酪氨酸酶活性、黑素含量的影响.方法 分别用0、10、20、30、40、50、100、200、300 mJ/cm2 BB-UVB照射原代培养的黑素细胞,采用CCK8法测定黑素细胞增殖率、多巴比色法测定酪氨酸酶活性、NaOH溶解法测定黑素含量.分别用0、30、50、100 mJ/cm2BB-UVB照射黑素细胞,实时荧光定量PCR检测非经典Wnt通路相关基因mRNA的表达.100 mJ/cm2 BB-UVB照射黑素细胞,用Western印迹检测作用前后非经典Wnt通路相关基因蛋白的表达.多组间比较采用单因素方差分析,两组间比较采用独立样本t检验.结果 与对照组相比,10 ~ 300 mJ/cm2 BB-UVB照射后,细胞增殖率均逐渐降低,BB-UVB剂量>100 mJ/cm2时细胞存活率<50%,同时,10~100 mJ/cm2 BB-UVB照射后酪氨酸酶活性逐渐增加,100 mJ/cm2 BB-UVB组黑素含量明显增加,差异有统计学意义(均P< 0.05).30、50、100 mJ/cm2BB-UVB照射后,WIF-1 mRNA的表达均较对照组逐渐减少,JNK、MITF、RAC1、TYR的表达均较对照组逐渐升高,而30、50 mJ/cm2 BB-UVB组WNT5A mRNA表达量均较对照组降低,100 mJ/cm2 BB-UVB组WNT5A mRNA表达量则明显升高(P<0.05).100 mJ/cm2 BB-UVB照射后,WIF-1蛋白的表达量较对照组降低,WNT5A、JNK、MITF、RAC1、TYR蛋白表达较对照组升高(P<0.05).结论 紫外线照射降低黑素细胞增殖率,提高黑素细胞酪氨酸酶活性和黑素含量.WIF-1基因可能抑制黑素生成.WIF-1基因表达降低可能通过非经典通路Wnt蛋白的综合作用激活JNK/MITF/TYR通路,最终促进黑素合成.%Objective To investigate the effects of broadband ultraviolet B (BB-UVB) on the proliferation of, tyrosinase activity and melanogenesis in melanocytes.Methods Melanocytes isolated from human foreskin were subjected to primary culture.Some cultured primary melanocytes were irradiated with BB-UVB at 10, 20, 30, 40, 50, 100, 200 and 300 mJ/cm2.Then

  13. Mutations in gfpt1 and skiv2l2 cause distinct stage-specific defects in larval melanocyte regeneration in zebrafish.

    Directory of Open Access Journals (Sweden)

    Chao-Tsung Yang

    2007-06-01

    Full Text Available The establishment of a single cell type regeneration paradigm in the zebrafish provides an opportunity to investigate the genetic mechanisms specific to regeneration processes. We previously demonstrated that regeneration melanocytes arise from cell division of the otherwise quiescent melanocyte precursors following larval melanocyte ablation with a small molecule, MoTP. The ease of ablating melanocytes by MoTP allows us to conduct a forward genetic screen for mechanisms specific to regeneration from such precursors or stem cells. Here, we reported the identification of two mutants, eartha(j23e1 and julie(j24e1 from a melanocyte ablation screen. Both mutants develop normal larval melanocytes, but upon melanocyte ablation, each mutation results in a distinct stage-specific defect in melanocyte regeneration. Positional cloning reveals that the eartha(j23e1 mutation is a nonsense mutation in gfpt1 (glutamine:fructose-6-phosphate aminotransferase 1, the rate-limiting enzyme in glucosamine-6-phosphate biosynthesis. Our analyses reveal that a mutation in gfpt1 specifically affects melanocyte differentiation (marked by melanin production at a late stage during regeneration and that gfpt1 acts cell autonomously in melanocytes to promote ontogenetic melanocyte darkening. We identified that the julie(j24e1 mutation is a splice-site mutation in skiv2l2 (superkiller viralicidic activity 2-like 2, a predicted DEAD-box RNA helicase. Our in situ analysis reveals that the mutation in skiv2l2 causes defects in cell proliferation, suggesting that skiv2l2 plays a role in regulating melanoblast proliferation during early stages of melanocyte regeneration. This finding is consistent with previously described role for cell division during larval melanocyte regeneration. The analyses of these mutants reveal their stage-specific roles in melanocyte regeneration. Interestingly, these mutants identify regeneration-specific functions not only in early stages of the

  14. Gene expression analysis of zebrafish melanocytes, iridophores, and retinal pigmented epithelium reveals indicators of biological function and developmental origin.

    Science.gov (United States)

    Higdon, Charles W; Mitra, Robi D; Johnson, Stephen L

    2013-01-01

    In order to facilitate understanding of pigment cell biology, we developed a method to concomitantly purify melanocytes, iridophores, and retinal pigmented epithelium from zebrafish, and analyzed their transcriptomes. Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co-enrichment in melanocytes and retinal pigmented epithelium, as well as in melanocytes and iridophores. We found 214 genes co-enriched in melanocytes and retinal pigmented epithelium, indicating the shared functions of melanin-producing cells. We found 62 genes significantly co-enriched in melanocytes and iridophores, illustrative of their shared developmental origins from the neural crest. This is also the first analysis of the iridophore transcriptome. Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine-based reflective pigment. We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate-limiting substrate for purine synthesis, phosphoribosyl pyrophosphate, thus constituting a guanine cycle. The purification procedure and expression analysis described here, along with the accompanying transcriptome-wide expression data, provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types, and will be a useful resource for further studies of pigment cell biology.

  15. A comparative study of mitochondrial ultrastructure in melanocytes from perilesional vitiligo skin and perilesional halo nevi skin.

    Science.gov (United States)

    Ding, Gao-Zhong; Zhao, Wen-E; Li, Xue; Gong, Qing-Li; Lu, Yan

    2015-04-01

    Vitiligo and halo nevi are both pigmentary disorders of the skin characterized by the acquired loss of functional epidermal melanocytes manifesting as white macules and patches. The cellular mechanism(s) and biochemical changes that result in the appearance of these two types of achromic lesions are still uncertain; and the relationship between vitiligo and halo nevi has been in dispute. In this study, we investigated the ultrastructure of mitochondria in melanocytes and in keratinocytes from perilesional vitiligo skin and from perilesional halo nevi skin using Transmission Electron Microscopy. Furthermore, we performed a quantitative analysis of mitochondrial morphology through a stereological study. As previously reported, we found that melanocytes from perilesional active vitiligo skin were loosely connected with their surroundings by their retracted dendrites. The surface density and the volume density of mitochondria in melanocytes and in keratinocytes from perilesional vitiligo skin are increased significantly compared with the controls, especially in active vitiligo. In contrast, there are no significant differences in mitochondria in melanocytes and in keratinocytes from perilesional halo nevi skin compared with the controls. In summary, the tendency of different morphologic alterations in mitochondria from perilesional vitiligo skin and from perilesional halo nevi skin reflect heterogeneous backgrounds between the two diseases, revealing that vitiligo and halo nevi may have separate pathogenic mechanisms. These findings may help elucidate the relationship of these two diseases and their underlying mechanisms.

  16. Gene expression analysis of zebrafish melanocytes, iridophores, and retinal pigmented epithelium reveals indicators of biological function and developmental origin.

    Directory of Open Access Journals (Sweden)

    Charles W Higdon

    Full Text Available In order to facilitate understanding of pigment cell biology, we developed a method to concomitantly purify melanocytes, iridophores, and retinal pigmented epithelium from zebrafish, and analyzed their transcriptomes. Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co-enrichment in melanocytes and retinal pigmented epithelium, as well as in melanocytes and iridophores. We found 214 genes co-enriched in melanocytes and retinal pigmented epithelium, indicating the shared functions of melanin-producing cells. We found 62 genes significantly co-enriched in melanocytes and iridophores, illustrative of their shared developmental origins from the neural crest. This is also the first analysis of the iridophore transcriptome. Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine-based reflective pigment. We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate-limiting substrate for purine synthesis, phosphoribosyl pyrophosphate, thus constituting a guanine cycle. The purification procedure and expression analysis described here, along with the accompanying transcriptome-wide expression data, provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types, and will be a useful resource for further studies of pigment cell biology.

  17. In vitro effect of heat treatment on the secretion of α-melanocyte-stimulating hormone by human melanocytes and HaCaT cells%热处理对体外人黑素细胞及HaCaT细胞分泌α黑素细胞刺激素的影响

    Institute of Scientific and Technical Information of China (English)

    祃丽娟; 赵广; 顾伟杰; 牛建荣; 邵丽芳; 孟如松; 闫文厅; 成玉

    2013-01-01

    目的 探讨热处理后体外培养的人表皮黑素细胞和人角质形成细胞(HaCaT细胞)分泌α黑素细胞刺激素(α-MSH)的变化.方法 将体外培养的黑素细胞及HaCaT细胞分别给予热处理(42℃,每天60 min,连续3d),实时荧光定量PCR法(RT-PCR)及酶联免疫吸附测定法(ELISA)检测两种细胞分泌的α-MSH的变化.结果 黑素细胞热处理组(1.5862±0.3262)与对照组(1.0000±0.2715)相比,α-MSH mRNA表达均明显增加(P<0.05);HaCaT细胞热处理组(1.8013±0.1216)与对照组(1.0000±0.2532)相比,mRNA水平也有明显增加(P<0.05).ELISA结果:黑素细胞热处理组(0.3142±0.0469)蛋白表达水平明显高于对照组(0.2557±0.0330,P< 0.05);HaCaT细胞热处理组(0.4632±0.0345)蛋白水平也明显高于对照组(0.4389±0.0399,P< 0.05).结论 热处理可促进人黑素细胞及HaCaT细胞α-MSH分泌.%Objetive To evaluate the in vitro effect of heat treatment on the secretion of α-melanocytestimulating hormone (α-MSH) by human melanocytes and HaCaT keratinocytes.Methods Cultured normal human melanocytes from forskin tissue and HaCaT cells were treated with hyperthermia at 42 ℃ for 1 hour per day for 3 days.Subsequently,real time fluorescence-based quantitative PCR and enzyme-linked immunosorbent assay (ELISA) were performed to detect the mRNA and protein expressions of α-MSH,respectively.Results The mRNA expression level of α-MSH was significantly higher in heat-treated melanocytes and HaCaT cells than in those untreated (1.5862 ± 0.3262 vs.1.0000 ± 0.2715,P < 0.05; 1.8013 ± 0.1216 vs.1.0000 ± 0.2532,P < 0.05).Moreover,ELISA showed a statistical increase in the supematant level of α-MSH protein in heat-treated melanocytes and HaCaT cells compared with those untreated (0.3142 ± 0.0469 vs.0.2557 ± 0.0330,P < 0.05; 0.4632 ± 0.0345 vs.0.4389 ± 0.0399,P < 0.05).Conclusion Heat treatment can promote the secretion of α-MSH by melanocytes and HaCaT cells.

  18. Isomerically Pure Tetramethylrhodamine Voltage Reporters.

    Science.gov (United States)

    Deal, Parker E; Kulkarni, Rishikesh U; Al-Abdullatif, Sarah H; Miller, Evan W

    2016-07-27

    We present the design, synthesis, and application of a new family of fluorescent voltage indicators based on isomerically pure tetramethylrhodamines. These new Rhodamine Voltage Reporters, or RhoVRs, use photoinduced electron transfer (PeT) as a trigger for voltage sensing, display excitation and emission profiles in the green to orange region of the visible spectrum, demonstrate high sensitivity to membrane potential changes (up to 47% ΔF/F per 100 mV), and employ a tertiary amide derived from sarcosine, which aids in membrane localization and simultaneously simplifies the synthetic route to the voltage sensors. The most sensitive of the RhoVR dyes, RhoVR 1, features a methoxy-substituted diethylaniline donor and phenylenevinylene molecular wire at the 5'-position of the rhodamine aryl ring, exhibits the highest voltage sensitivity to date for red-shifted PeT-based voltage sensors, and is compatible with simultaneous imaging alongside green fluorescent protein-based indicators. The discoveries that sarcosine-based tertiary amides in the context of molecular-wire voltage indicators prevent dye internalization and 5'-substituted voltage indicators exhibit improved voltage sensitivity should be broadly applicable to other types of PeT-based voltage-sensitive fluorophores.

  19. Evaluation of various pesticides-degrading pure bacterial cultures ...

    African Journals Online (AJOL)

    IASA

    2016-10-05

    Oct 5, 2016 ... that the highest growth rate of microbial consortium was observed during degradation of various pesticides .... is Gram positive, aerobic, catalase, oxidase and King B .... sp. nov., isolated from root nodules of Pisumsativum.

  20. Protective effects of tea polyphenols against the destruction of melanocytes by CD8+ T ceils from vitiligo patients%茶多酚对CD8+T细胞杀伤白癜风患者黑素细胞的保护作用研究

    Institute of Scientific and Technical Information of China (English)

    欧阳杰; 吴纪龙; 周妙妮; 付丽芳; 许爱娥

    2013-01-01

    Objective To evaluate the protective effects of tea polyphenols against the destruction of melanocytes by CD8+ T cells from vitiligo patients.Methods Skin tissue was resected from the margin of vitiligo lesions followed by the isolation and culture of CD8+ T lymphocytes,and from the normal skin of vitiligo patients followed by the isolation and culture of melanocytes.Flow cytometry was carried out to evaluate the purity of CD8+ T cells.The melanocytes were cocultured with the CD8 + T cells at different ratios followed by the evaluation of killing effect of CD8+ T cells.Various concentrations (200 and 400 μg/ml) of tea polyphenols were added into the co-culture system of CD8+ T cells and melanocytes at a ratio of 5 ∶ 1 followed by an additional culture of 48 hours.Then,flow cytometry was performed to detect the apoptosis in melanocytes in the coculture system.Results CD8+ T lymphocytes were successfully obtained from the marginal area of vitiligo lesions with a purity of more than 90%,which highly expressed the antigens CD137 and CD69.The coculture with CD8+ T cells markedly accelerated the apoptosis in melanocytes,while the accelerative effect was inhibited by tea polyphenols of 200 and 400 μg/ml.Conclusions The CD8+ T cells infiltrating the edge of vitiligo lesions display a potential destructive effect on autologous melanocytes from vitiligo patients,and tea polyphenols have a protective effect against the destruction of melanocytes by CD8+ T cells.%目的 探讨茶多酚对CD8+T细胞杀伤白癜风患者黑素细胞的保护作用.方法 取白癜风患者白斑边缘处皮片组织,培养CD8+T淋巴细胞,细胞纯度用流式细胞仪进行鉴定,通过和黑素细胞共培养验证其针对黑素细胞的杀伤作用.将不同浓度茶多酚加入CD8+T细胞和黑素细胞共培养的体系,用流式细胞仪检测黑素细胞的凋亡.结果 取白癜风皮损边缘表皮组织中成功培养CD8+T淋巴细胞,纯度达90%

  1. The Influence of Ferulic Acid Solutions on Proliferation of Melanocytes at Different Con-centrations%不同浓度阿魏酸溶液对人表皮黑素细胞增殖的影响

    Institute of Scientific and Technical Information of China (English)

    汤楠; 吴艳华; 蔡兰花; 李其林

    2014-01-01

    Objective:To investigate the impact of ferulic acid on proliferation of epidermal melanocytes in vitro.Methods:The normal epidermal melanocytes were identified by L-DOPA staining.After being treated with different concentrations (0.01,0.1,1 mg/mL) of ferulic acid for 24, 48, 72 h in vitro, the proliferative activity of cultured melanocytes were detected by MTS . Results:Compared with the control group without treatment with ferulic acid , the proliferation of the melanocytes were inhibited when treated with 0.01, 0.1, 1 mg/mL ferulic acid.The differ-ence were statistically significant ( P<0 .05 ) .Concentration and time have interaction .With the higher concentration and the prolonged of culture time , the inhibition of melanocyte proliferation was enhanced .Conclusion:Ferulic acid solution may inhibit the proliferation of normal human epi-dermal melanocytes .It suggests that ferulic acid may have potentials in the treatment of melasma and other pigmented disorders .%目的:观察不同浓度阿魏酸溶液对体外培养的正常人表皮黑素细胞增殖的影响。方法:采用L-DOPA染色法鉴定正常人表皮黑素细胞;分别用0.01、0.1、1 mg/mL的阿魏酸溶液处理正常人表皮黑素细胞24 h、48 h、72 h,采用MTS法测定黑素细胞增殖活性。结果:与未用阿魏酸溶液处理的对照组相比,0.01、0.1、1 mg/mL阿魏酸在作用24、48、72 h后,黑素细胞增殖作用均受到抑制,差异均有统计学意义(P值均<0.05);不同浓度与时间有交互作用(F=2.48,P=0.034),随着阿魏酸溶液浓度的增加及作用时间的延长,对黑素细胞的抑制作用增强。结论:阿魏酸溶液可以抑制正常人表皮黑素细胞的增殖,提示其在治疗黄褐斑等疾病方面具有良好的前景。

  2. Influence of melanocytes in the ex-vivo reconstructed epidermal melanin unit following an acute UV irradiation; Role des melanocytes dans l'unite epidermique de melanisation reconstruite ex-vivo apres une irradiation UV aigue

    Energy Technology Data Exchange (ETDEWEB)

    Cario-Andre, M

    2000-11-15

    Influence of melanocytes in skin pigmentation is well documented, however its photo-protective role has given rise to controversy. The role of melanocytes have been investigated on reconstructed epidermis with 100 % of keratinocytes or 95 % of keratinocytes and 5 % of melanocytes. In a first time, the effect of an acute UVB dose has been studied on both reconstructed epidermis, next we have investigated UVA and UVA+B effects on these epidermis. Following irradiation, the presence of melanocytes in reconstructed epidermis protects against apoptosis without protecting significantly against DNA damage formation (CPD, 6-4PP) and protects against UV-induced unbalance of the SOD/catalase ratio (antioxidants enzymes). On the contrary, the presence of melanocytes in reconstructed epidermis amplifies lipids and proteins oxidations but seems to protect against DNA oxidations. Melanocytes differ from keratinocytes by their melanin content and their more important concentration in polyunsaturated fatty acids. To evaluate what is the part of melanin and the part of polyunsaturated fatty acids in epidermal UV responses, reconstructed epidermis with keratinocytes have been supplemented with polyunsaturated fatty acid. This study indicates that polyunsaturated fatty acids are responsible for lipids and proteins oxidations and that melanin protect against DNA oxidation induced by lipid peroxidation. All these studies demonstrate that, model of reconstructed epidermis and epidermis in-vivo have the same behaviour following UV irradiation. In the last part, sunscreens and antioxidants have been tested on reconstructed epidermis and have demonstrated that model of reconstructed epidermis is suitable for photo-protective molecules screening. (author)

  3. MicroRNA expression in melanocytic nevi: the usefulness of formalin-fixed, paraffin-embedded material for miRNA microarray profiling

    DEFF Research Database (Denmark)

    Glud, Martin; Klausen, Mikkel; Gniadecki, Robert

    2009-01-01

    surgical specimens are formalin fixed and paraffin embedded (FFPE). To explore whether FFPE material would be suitable for miRNA profiling in melanocytic lesions, we compared miRNA expression patterns in FFPE versus fresh frozen samples, obtained from 15 human melanocytic nevi. Out of microarray data, we...

  4. Spindle cell melanocytic lesions--part I: an approach to compound naevoidal pattern lesions with spindle cell morphology and Spitzoid pattern lesions.

    Science.gov (United States)

    Sade, Shachar; Al Habeeb, Ayman; Ghazarian, Danny

    2010-04-01

    Melanocytic lesions show great morphological diversity in their architecture and the cytomorphological appearance of their composite cells. Whereas functional melanocytes reveal a dendritic cytomorphology and territorial isolation, lesional naevomelanocytes and melanoma cells typically show epithelioid, spindled or mixed cytomorphologies and a range of architectural arrangements. Spindling is common to melanocytic lesions, and may be either a characteristic feature or a divergent appearance. The presence of spindle cells may mask the melanocytic nature of a lesion, and is often disconcerting, either because of its infrequent appearance in a particular lesion or its interpretation as a dedifferentiated phenotype. Spindle cell melanocytic lesions follow the full spectrum of potential biological outcomes, and difficulty may be experienced judging the nature of a lesion because of a lack of consistently reliable features to predict biological behaviour. Over time, recognition of numerous histomorphological features that may portend a more aggressive lesion have been identified. However, the translation of these features into a diagnostic entity requires a gestalt approach. Although most spindle cell melanocytic lesions can reliably be resolved with this standard approach, problem areas do exist and cause no end of grief to the surgical pathologist or dermatopathologist. In this review, the authors present their algorithmic approach to spindle cell melanocytic lesions and discuss each entity in turn, in order to (1) model a systematic approach to such lesions, and (2) provide familiarity with those melanocytic lesions that either typically or occasionally display a spindled cytomorphology.

  5. Pure variation and organic stratification.

    Science.gov (United States)

    Rosanvallon, Jérôme

    2012-09-01

    The fundamental problem posed by Darwin distinguishes his theory from any transformism of the past as well as any evolutionism to come: since variation is inherent to the living, it is a question of explaining, not at all why the living varies, but instead why the living does not vary in all directions to the point of constituting a continuum of forms varying ad infinitum. What limits and stabilizes this intrinsically unlimited variation, allowing certain forms to subsist and multiply to the detriment of others, is natural selection. This double principle of intrinsic variation/extrinsic selection constitutes a vector for the unification of reality that underlies Jean-Jacques Kupiec's ontophylogenesis as well as Deleuze and Guattari's global philosophy of Nature. Therefore, everything would potentially tend to incessantly vary. The work of Kupiec and others identifies an intrinsic random variation within ontogenesis itself. For Deleuze and Guattari, it is nothing but the figure, already selected by the organic stratum, of a more fundamental or pure variation. But, in fact, nothing really varies incessantly: everything undergoes a selective pressure according to which nothing subsists as such except what manages to endure through invariance (physical stratum) or reproduction (organic stratum). Thus, organic stratification only retains from variation what ensures and augments this reproduction. In this sense, every organism stratifies, i.e. submits to its imperative of subsistence and reproduction, a body without organs that varies in itself and always tends to escape the organism, for better (intensifications of life) or worse (cancerous pathologies). Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. 甘草提取物对人黑素细胞黑素合成的影响%The impact of licorice on the melanin synthesis of human epidermal primary melanocytes

    Institute of Scientific and Technical Information of China (English)

    岳学状; 王大光; 朱文元

    2011-01-01

    目的: 评价甘草提取物对人原代黑素细胞酪氨酸酶及黑素合成的影响.方法:选择2.5、5.0、10.0、15.0、20.0、30.0 mg/L6个浓度的甘草作用于体外培养的人表皮黑素细胞72 h,分别测定细胞增殖活性、酪氨酸酶活性和黑素含量,western blot方法检测甘草作用后黑素细胞中酪氨酸酶蛋白的表达.结果: 甘草显著抑制黑素合成和酪氨酸酶活性,且呈浓度依赖性.结论: 甘草通过下调酪氨酸酶的表达抑制黑素合成和酪氨酸酶活性.%Objective: To assess the effects of licorice on melanin synthesis. Methods: Normal human melanocytes were isolated from human forskin and incubated with 2.5, 5.0, 10.0, 15.0, 20.0 and 30.0 fig/mL licorice for 72 h respectively, in vivo. The proliferation of melanocytes, melanin content and tyrosinase activity were measured. The protein expression of tyrosinase was evaluated with western blot technique. Results: Melanin synthesis was reduced and tyrosinase activity was significantly inhibited when licorice was cultured with melanocytes in a dose dependent manner, compared to control group. Conclusion: Licorice can inhibit melanogenesis and tyrosinase activity through downregulating the expression of tyrosinase.

  7. Effects of ginsenoside Rb1 on melanogenesis in human epidermal melanocytes%人参皂苷Rb1对人表皮黑素细胞黑素生成的影响

    Institute of Scientific and Technical Information of China (English)

    卢珊珊; 李国艳; 赵丹; 张微; 林茂; 涂彩霞

    2012-01-01

    from circumcision specimens of children,and subjected to primary culture.After 2 to 5 passages,the melanocytes were treated with different concentrations of ginsenoside Rb1,dimethyl sulfoxide (DMSO,vehicle control),forskolin at 10 μmol/L(positive control) or remained untreated (blank control).After additional culture for 72 hours,methyl thiazolyl tetrazolium (MTT) assay and NaOH lysis method were used to evaluate cell viability and melanin content in melanocytes respectively,spectrophotometer to determine dopa oxidase activity of tyrosinase,Western blot to quantify the protein level of tyrosinase,microphthalmia-associated transcription factor (MITF),phosphorylated and total cAMP response element binding protein (p-CREB and t-CREB) in melanocytes.[Results] After treatment with ginsenoside Rbl of 25,50 and 100 μmol/L for 72 hours,the melanocytes experienced no significant changes in viability (P > 0.05 ),but a significant dose-dependent increase in melanin content (112.4%± 5.7%,155.7% + 6.3%,217.2% ± 11.7% vs.100%,P< 0.05 or 0.01) and tyrosinase activity(117.9% ± 5.7%,158.2% ± 9.6%,182.6% ± 10.0% vs.100%,P< 0.05 or 0.01 ) compared with the vehicle control melanocytes.The protein expressions of tyrosinase,MITF and p-CREB were statistically higher in melanocytes treated with ginsenoside Rb1 of 100 μmol/L for 72 hours than in the vehicle control melanocytes (225.4% ± 12.8% vs.100% ± 7.9%,313.5% ± 16.7% vs.100% ± 9.8%,322.5% ± 21.1% vs.100% ± 9.1%,all P< 0.01).The increase in MITF protein expression was inapparent in melanocytes at 8 hours after the treatment with ginsenoside Rb1 of 100 μmol/L,but statistically significant at 24 hours compared with the melanocytes at baseline (P< 0.01).The pretreatment with H-89 (a 8elective inhibitor of PKA) at 10 μmol/L,significantly suppressed the ginsenoside Rb1 (100 μmol/L for 72 hours) -induced phosphorylation of CREB,increase in MITF,tyrosinase expression

  8. Transcriptional and signaling regulation in neural crest stem cell-derived melanocyte development:do all roads lead to Mitf?

    Institute of Scientific and Technical Information of China (English)

    Ling Hou; William J Pavan

    2008-01-01

    Human neurocristopathies include a number of syndromes,tumors,and dysmorphologies of neural crest (NC) stem cell derivatives.In recent years,many white spotting genes have been associated with hypopigmentary disorders and deafness in neurocristopathies resulting from NC stem cell-derived melanocyte deficiency during development.These include PAX3,SOX10,MITF,SNAI2,EDNRB,EDN3,KIT,and KITL.Recent studies have revealed surprising new insights into a central role of MITF in the complex network of interacting genes in melanocyte development.In this perspective,we provide an overview of some of the current findings and explore complex functional roles of these genes during NC stem cell-derived melanocyte development.

  9. Mutations in c10orf11, a melanocyte-differentiation gene, cause autosomal-recessive albinism.

    Science.gov (United States)

    Grønskov, Karen; Dooley, Christopher M; Østergaard, Elsebet; Kelsh, Robert N; Hansen, Lars; Levesque, Mitchell P; Vilhelmsen, Kaj; Møllgård, Kjeld; Stemple, Derek L; Rosenberg, Thomas

    2013-03-07

    Autosomal-recessive albinism is a hypopigmentation disorder with a broad phenotypic range. A substantial fraction of individuals with albinism remain genetically unresolved, and it has been hypothesized that more genes are to be identified. By using homozygosity mapping of an inbred Faroese family, we identified a 3.5 Mb homozygous region (10q22.2-q22.3) on chromosome 10. The region contains five protein-coding genes, and sequencing of one of these, C10orf11, revealed a nonsense mutation that segregated with the disease and showed a recessive inheritance pattern. Investigation of additional albinism-affected individuals from the Faroe Islands revealed that five out of eight unrelated affected persons had the nonsense mutation in C10orf11. Screening of a cohort of autosomal-recessive-albinism-affected individuals residing in Denmark showed a homozygous 1 bp duplication in C10orf11 in an individual originating from Lithuania. Immunohistochemistry showed localization of C10orf11 in melanoblasts and melanocytes in human fetal tissue, but no localization was seen in retinal pigment epithelial cells. Knockdown of the zebrafish (Danio rerio) homolog with the use of morpholinos resulted in substantially decreased pigmentation and a reduction of the apparent number of pigmented melanocytes. The morphant phenotype was rescued by wild-type C10orf11, but not by mutant C10orf11. In conclusion, we have identified a melanocyte-differentiation gene, C10orf11, which when mutated causes autosomal-recessive albinism in humans.

  10. Knowledge building insights on biomarkers of arsenic toxicity to keratinocytes and melanocytes.

    Science.gov (United States)

    Isokpehi, Raphael D; Udensi, Udensi K; Anyanwu, Matthew N; Mbah, Andreas N; Johnson, Matilda O; Edusei, Kafui; Bauer, Michael A; Hall, Roger A; Awofolu, Omotayo R

    2012-01-01

    Exposure to inorganic arsenic induces skin cancer and abnormal pigmentation in susceptible humans. High-throughput gene transcription assays such as DNA microarrays allow for the identification of biological pathways affected by arsenic that lead to initiation and progression of skin cancer and abnormal pigmentation. The overall purpose of the reported research was to determine knowledge building insights on biomarker genes for arsenic toxicity to human epidermal cells by integrating a collection of gene lists annotated with biological information. The information sets included toxicogenomics gene-chemical interaction; enzymes encoded in the human genome; enriched biological information associated with genes; environmentally relevant gene sequence variation; and effects of non-synonymous single nucleotide polymorphisms (SNPs) on protein function. Molecular network construction for arsenic upregulated genes TNFSF18 (tumor necrosis factor [ligand] superfamily member 18) and IL1R2 (interleukin 1 Receptor, type 2) revealed subnetwork interconnections to E2F4, an oncogenic transcription factor, predominantly expressed at the onset of keratinocyte differentiation. Visual analytics integration of gene information sources helped identify RAC1, a GTP binding protein, and TFRC, an iron uptake protein as prioritized arsenic-perturbed protein targets for biological processes leading to skin hyperpigmentation. RAC1 regulates the formation of dendrites that transfer melanin from melanocytes to neighboring keratinocytes. Increased melanocyte dendricity is correlated with hyperpigmentation. TFRC is a key determinant of the amount and location of iron in the epidermis. Aberrant TFRC expression could impair cutaneous iron metabolism leading to abnormal pigmentation seen in some humans exposed to arsenicals. The reported findings contribute to insights on how arsenic could impair the function of genes and biological pathways in epidermal cells. Finally, we developed visual analytics

  11. Premalignant quiescent melanocytic nevi do not express the MHC class I chain-related protein A

    Directory of Open Access Journals (Sweden)

    Mercedes B. Fuertes

    2011-08-01

    Full Text Available The MHC class I chain-related protein A (MICA is an inducible molecule almost not expressed by normal cells but strongly up-regulated in tumor cells. MICA-expressing cells are recognized by natural killer (NK cells, CD8+ aßTCR and ?dTCR T lymphocytes through the NKG2D receptor. Engagement of NKG2D by MICA triggers IFN-? secretion and cytotoxicity against malignant cells. Although most solid tumors express MICA and this molecule is a target during immune surveillance against tumors, it has been observed that high grade tumors from different histotypes express low amounts of cell surface MICA due to a metalloprotease- induced shedding. Also, melanomas develop after a complex process of neotransformation of normal melanocytes. However, the expression of MICA in premalignant stages (primary human quiescent melanocytic nevi remains unknown. Here, we assessed expression of MICA by flow cytometry using cell suspensions from 15 primary nevi isolated from 11 patients. When collected material was abundant, cell lysates were prepared and MICA expression was also analyzed by Western blot. We observed that MICA was undetectable in the 15 primary nevi (intradermic, junction, mixed, lentigo and congenital samples as well as in normal skin, benign lesions (seborrheic keratosis, premalignant lesions (actinic keratosis and benign basocellular cancer. Conversely, a primary recently diagnosed melanoma showed intense cell surface MICA. We conclude that the onset of MICA expression is a tightly regulated process that occurs after melanocytes trespass the stage of malignant transformation. Thus, analysis of MICA expression in tissue sections of skin samples may constitute a useful marker to differentiate between benign and malignant nevi.

  12. Premalignant quiescent melanocytic nevi do not express the MHC class I chain-related protein A.

    Science.gov (United States)

    Fuertes, Mercedes B; Rossi, Lucas E; Peralta, Carlos M; Cabrera, Hugo N; Allevato, Miguel A; Zwirner, Norberto W

    2011-01-01

    The MHC class I chain-related protein A (MICA) is an inducible molecule almost not expressed by normal cells but strongly up-regulated in tumor cells. MICA-expressing cells are recognized by natural killer (NK) cells, CD8+ abTCR and gdTCR T lymphocytes through the NKG2D receptor. Engagement of NKG2D by MICA triggers IFN-g secretion and cytotoxicity against malignant cells. Although most solid tumors express MICA and this molecule is a target during immune surveillance against tumors, it has been observed that high grade tumors from different histotypes express low amounts of cell surface MICA due to a metalloprotease-induced shedding. Also, melanomas develop after a complex process of neotransformation of normal melanocytes. However, the expression of MICA in premalignant stages (primary human quiescent melanocytic nevi) remains unknown. Here, we assessed expression of MICA by flow cytometry using cell suspensions from 15 primary nevi isolated from 11 patients. When collected material was abundant, cell lysates were prepared and MICA expression was also analyzed by Western blot. We observed that MICA was undetectable in the 15 primary nevi (intradermic, junction, mixed, lentigo and congenital samples) as well as in normal skin, benign lesions (seborrheic keratosis), premalignant lesions (actinic keratosis) and benign basocellular cancer. Conversely, a primary recently diagnosed melanoma showed intense cell surface MICA. We conclude that the onset of MICA expression is a tightly regulated process that occurs after melanocytes trespass the stage of malignant transformation. Thus, analysis of MICA expression in tissue sections of skin samples may constitute a useful marker to differentiate between benign and malignant nevi.

  13. Tame Kernels of Pure Cubic Fields

    Institute of Scientific and Technical Information of China (English)

    Xiao Yun CHENG

    2012-01-01

    In this paper,we study the p-rank of the tame kernels of pure cubic fields.In particular,we prove that for a fixed positive integer m,there exist infinitely many pure cubic fields whose 3-rank of the tame kernel equal to m.As an application,we determine the 3-rank of their tame kernels for some special pure cubic fields.

  14. Dissipation-induced pure Gaussian state

    CERN Document Server

    Koga, Kei

    2011-01-01

    This paper provides some necessary and sufficient conditions for a general Markovian Gaussian master equation to have a unique pure steady state. The conditions are described by simple matrix equations, thus they can be easily applied to the so-called environment engineering for pure Gaussian state preparation. In particular, it is shown that for any given pure Gaussian state we can actually construct a dissipative process yielding that state as the unique steady state.

  15. Intracranial melanocytic meningeal tumours and melanosis oculi: case report and literature review

    Directory of Open Access Journals (Sweden)

    Doglietto Francesco

    2012-06-01

    Full Text Available Abstract Background Melanocytic meningeal tumours are rare extra-axial neoplasms of the nervous system, with only three reported cases in the cavernous sinus. Herein we describe for the first time the association of ocular melanosis and multiple intracranial melanocytic meningeal tumours, with the presenting lesion being in the cavernous sinus. The importance of this association is discussed together with the diagnostic and therapeutic challenges of the case. Case presentation A 20-year-old man presented with a left sixth cranial nerve deficit; general examination documented only congenital melanosis of the homolateral eye. MRI examination showed a space occupying lesion in the left cavernous sinus, which was followed conservatively for 2 years, until a new space occupying lesion was evident at the level of the right frontal convexity: both lesions presented with neuroradiological characteristics suggestive of melanin content. The frontal convexity lesion was removed: intraoperatively the dura was markedly and diffusely melanotic. Histological examination documented a melanocytic meningeal tumour, with a proliferative index of 3 %. The patient underwent 3D-Conformal Radiation Therapy on the lesion of the cavernous sinus (total dose 5040 cGy, with initial tumour reduction. Three years later, due to a symptomatic growth, he underwent partial removal of the lesion in the cavernous sinus. Histological examination was unchanged. He then received adjuvant Temozolomide with Low Dose Fractionated Radiation Therapy (LD-FRT. Due to further disease progression cisplatin plus fotemustine were administered, concomitant with LD-FRT: after two cycles MRI documented significant disease regression. After a period of apparent disease control, the patient presented with persistent cough and evidence of multiple thoracic metastases, which lead to his death, seven years after presentation. Conclusions Intracranial melanocytic meningeal tumours are challenging

  16. 白癜风皮损中非黑素细胞进展%Non-melanocytes in vitiligo lesions

    Institute of Scientific and Technical Information of China (English)

    金永红; 许爱娥

    2008-01-01

    The pathogenesis of vitiligo is still unclear.In addition to melanocytes,non-melanocytes are involved in the pathogenesis of vitiligo.More and more studies have been conducted on non.melanocytes in vitiligo recently.It has been shown that keratinocytes,Langerhans cells and fibroblasts affect the migration,proliteration and differentiation of melanocytes.The changes of cytokines secreted by and ultrastructural abnormality of non-melanocytes may affect the activity and apoptosis of melanocytes in vitiligo lesions,influence pigrnentogenesis in skin,thereafter,take part in the pathogenesis of vitiligo.This paper presents the latest investigation of keratinocytes,Langerhans cells and fibroblasts in vitiligo.%白癜风的发病机制尚未完全明了.其发病除黑素细胞异常外,还涉及非黑素细胞的变化,近年来非黑素细胞在白癜风中的研究逐渐增多.研究表明,一些皮肤非黑素细胞如角质形成细胞、朗格汉斯细胞、成纤维细胞等与黑素细胞关系密切,这些细胞影响黑素细胞的迁移、增殖、分化等功能.白癜风皮损中非黑素细胞超微结构的异常和分泌细胞因子的变化可影响黑素细胞的活性及凋亡,影响皮肤色素生成,从而参与白癜风发病.

  17. MicroRNA-27a-3p Inhibits Melanogenesis in Mouse Skin Melanocytes by Targeting Wnt3a.

    Science.gov (United States)

    Zhao, Yuanyuan; Wang, Pengchao; Meng, Jinzhu; Ji, Yuankai; Xu, Dongmei; Chen, Tianzhi; Fan, Ruiwen; Yu, Xiuju; Yao, Jianbo; Dong, Changsheng

    2015-05-14

    MicroRNAs (miRNAs) play an essential role in the regulation of almost all the biological processes, including melanogenesis. MiR-27a-3p is nearly six times higher in white alpaca skin compared to brown skin, which indicates that miR-27a-3p may be a candidate regulator for melanogenesis. Wnt3a plays an important role in promoting melanoblasts to differentiate into melanocytes and melanogenesis. To confirm the function of miR-27a-3p to melanogenesis in mammals, miR-27a-3p mimic, inhibitor and their negative control were transfected into mouse melanocytes. As a result, miR-27a-3p inhibits melanogenesis by repressing Wnt3a at post-transcriptional level. A significant decrease in Wnt3a luciferase activity was observed in 293T cells co-transfected with the matched luciferase reporter vector and pre-miR-27a. Furthermore, the presence of exogenous miR-27a-3p significantly decreased Wnt3a protein expression rather than mRNA and reduced β-catenin mRNA levels in melanocytes. The over-expression of miR-27a-3p significantly increased the melanin content of melanocytes. However, miR-27a-3p inhibitor performs an opposite effect on melanogenesis. Wnt3a is one target of miR-27a-3p. MiR-27a-3p could inhibit Wnt3a protein amount by post-transcriptional regulation and melanogenesis in mouse melanocytes. Previous studies reported that Wnt3a promoted melanogenensis in mouse melanocytes. Thus, miR-27-3p inhibits melanogenesis by repressing Wnt3a protein expression.

  18. Influence of TLR3 activation on expression of reactive oxygen species in melanocytes%TLR3活化对正常人表皮黑素细胞内活性氧簇表达的影响

    Institute of Scientific and Technical Information of China (English)

    于宁; 陆家睛; 易雪梅; 丁杨峰

    2015-01-01

    Objective To investigate the effect of Toll-like receptor 3 (TLR3) activation on the expression of reactive oxygen species ( ROS) in melanocytes .Methods Primary melanocytes were isolated from normal human skin and cultured in vitro, then they were inoculated into six pore plates .They were divided into the experimental group and the control group with 9 holes in each group.The cells in the experimental group were added with TLR 3 ligand poly (I∶C) to inoculate for 2, 6 and 24 hours, while the cells in the control group was not added .The expression of ROS was measured by CM-H2DCFDA probe method combined with flow cytometry .ResultsWith the extension of incubation time , the expression of ROS in the melanocytes was increased in the control group , but there was no statistically significant difference .With the extension of incubation time , the expression of ROS in the melanocytes was significantly increased in the experimental group (all P<0.01).At each time point, the expression of ROS in the melanocytes of the experimental group was higher than that of the control group (all P<0.01).Conclusion The TLR3 activator poly(I∶C) could induce the increase of ROS in the melanocytes .%目的:探讨Toll样受体3(TLR3)活化对黑素细胞内活性氧簇(ROS)表达的影响。方法分离并培养正常人表皮黑素细胞,接种于6孔板。设实验组和对照组,各9孔。实验组加入100 ng/mL TLR3激活剂poly(I∶C)孵育2、6、24 h,对照组不加药。用CM-H2DCFDA探针法联合流式细胞术检测黑素细胞内ROS表达量。结果随孵育时间延长,对照组黑素细胞内ROS表达量增加,但差异无统计学意义。实验组黑素细胞内ROS表达量随孵育时间延长显著增加(P均<0.01);在每一时间点,实验组黑素细胞内ROS表达量均高于相应的对照组(P均<0.01)。结论 TLR3激活可显著提高黑素细胞内ROS水平。

  19. The Expansion Postponement in Pure Type Systems

    Institute of Scientific and Technical Information of China (English)

    宋方敏

    1997-01-01

    The expansion postponement problem in Pure Type Systems is an open problem raised by R.Pollack in 1992.In this paper,the author presents a set of necessary and sufficient conditions for this problem and a set of sufficient conditions for it.The author also gives some properties for pure typ systems without the expansion rule.

  20. Measuring the entanglement of bipartite pure states

    CERN Document Server

    Sancho, J M

    2000-01-01

    The problem of the experimental determination of the amount of entanglement of a bipartite pure state is addressed. We show that measuring a single observable does not suffice to determine the entanglement of a given unknown pure state of two particles. Possible minimal local measuring strategies are discussed and a comparison is made on the basis of their best achievable precision.

  1. Fundamentals of the pure spinor formalism

    NARCIS (Netherlands)

    Hoogeveen, J.

    2010-01-01

    This thesis presents recent developments within the pure spinor formalism, which has simplified amplitude computations in perturbative string theory, especially when spacetime fermions are involved. Firstly the worldsheet action of both the minimal and the non-minimal pure spinor formalism is derive

  2. Oxidative Stress Modulates DNA Methylation during Melanocyte Anchorage Blockade Associated with Malignant Transformation

    Directory of Open Access Journals (Sweden)

    Ana C.E. Campos

    2007-12-01

    Full Text Available Both oxidative/nitrosative stress and alterations in DNA methylation are observed during carcinogenesis of different tumor types, but no clear correlation between these events has been demonstrated until now. Melanoma cell lines were previously established after submitting the nontumorigenic melanocyte lineage, melan-a, to cycles of anchorage blockade. In this work, increased intracellular oxidative species and nitric oxide levels, as well as alterations in the DNA methylation, were observed after melan-a detachment, which were also associated with a decrease in intracellular homocysteine (Hey, an element in the methionine (universal methyl donor cycle. This alteration was accompanied by increase in glutathione (GSH levels and methylated DNA content. Furthermore, a significant increase in dnmti and 3b expression was identified along melan-a anchorage blockade. LG-Nitro-L-arginine methyl esther (L-NAME, known as a nitric oxide synthase (NOS inhibitor, and N-acetyl-L-cysteine (NAC prevented the increase in global DNA methylation, as well as the increase in dnmti and 3b expression, observed during melan-a detachment. Interestingly, both L-NAME and NAC did not inhibit nitric oxide (NO production in these cells, but abrogated superoxide anion production during anchorage blockade. In conclusion, oxidative stress observed during melanocyte anchorage blockade seems to modulate DNA methylation levels and may directly contribute to the acquisition of an anoikis-resistant phenotype through an epigenetic mechanism.

  3. Patterns in melanocytic lesions: impact of the geometry on growth and transport inside the epidermis

    Science.gov (United States)

    Balois, Thibaut; Chatelain, Clément; Ben Amar, Martine

    2014-01-01

    In glabrous skin, nevi and melanomas exhibit pigmented stripes during clinical dermoscopic examination. They find their origin in the basal layer geometry which periodically exhibits ridges, alternatively large (limiting ridges) and thin (intermediate ridges). However, nevus and melanoma lesions differ by the localization of the pigmented stripes along furrows or ridges of the epidermis surface. Here, we propose a biomechanical model of avascular tumour growth which takes into account this specific geometry in the epidermis where both kinds of lesions first appear. Simulations show a periodic distribution of tumour cells inside the lesion, with a global contour stretched out along the ridges. In order to be as close as possible to clinical observations, we also consider the melanin transport by the keratinocytes. Our simulations show that reasonable assumptions on melanocytic cell repartition in the ridges favour the limiting ridges of the basal compared with the intermediate ones in agreement with nevus observations but not really with melanomas. It raises the question of cell aggregation and repartition of melanocytic cells in acral melanomas and requires further biological studies of these cells in situ. PMID:24872499

  4. Malignant Tumor Derived from Skin Melanocytes of a Bovine of Unusual Presentation: A Case Study

    Directory of Open Access Journals (Sweden)

    Carlos Alberto Chaves Velásquez

    2015-05-01

    Full Text Available Melanocytic tumors and melanomas in domestic animals include neoplasms composed of melanin-producing cells. In cattle, these tumors are rare and mostly benign, while malignant tumors are almost non-existent. The article reports the case of a female crossbred cow 38 months of age with a fluctuating mass located between the mandibular border and the left parotid region, about three months duration, with evident growth in the last thirty days. After surgical excision, a sample preserved in buffered formalin (10% was sent to the Laboratory of Pathology (University of Nariño—consisting of a fragment of 7.0 × 10.5 × 8.0 cm, ellipsoid, with skin and hair on one side, irregular surface, blackish brown, semi-soft consistency, and presence of shear translucent slimy content—for processing and inclusion in paraffin, cut to 5 μm thickness and stained with hematoxylin-eosin coloration. The forwarded tissue was classified as a neoplasm of malignant behavior derived from melanocytes, due to its cellular characteristics: growth pattern, pattern of distribution, severe cellular pleomorphism, anisocytosis, megalocytosis, nuclear pleomorphism, anisokaryosis, megalokaryosis, and involvement of blood vessel walls; additionally, the paraffin block was cut for immunohistochemical processing using monoclonal markers (S-100 DAKO® and Melan A DAKO®, contrasted with Mayer’s hematoxylin. Strong immunostaining of neoplastic cells is evident, and it constitutes the first reported case of this disease in Nariño (Colombia.

  5. Refractometry of melanocyte cell nuclei using optical scatter images recorded by digital Fourier microscopy.

    Science.gov (United States)

    Seet, Katrina Y T; Nieminen, Timo A; Zvyagin, Andrei V

    2009-01-01

    The cell nucleus is the dominant optical scatterer in the cell. Neoplastic cells are characterized by cell nucleus polymorphism and polychromism-i.e., the nuclei exhibits an increase in the distribution of both size and refractive index. The relative size parameter, and its distribution, is proportional to the product of the nucleus size and its relative refractive index and is a useful discriminant between normal and abnormal (cancerous) cells. We demonstrate a recently introduced holographic technique, digital Fourier microscopy (DFM), to provide a sensitive measure of this relative size parameter. Fourier holograms were recorded and optical scatter of individual scatterers were extracted and modeled with Mie theory to determine the relative size parameter. The relative size parameter of individual melanocyte cell nuclei were found to be 16.5+/-0.2, which gives a cell nucleus refractive index of 1.38+/-0.01 and is in good agreement with previously reported data. The relative size parameters of individual malignant melanocyte cell nuclei are expected to be greater than 16.5.

  6. A novel BAP1 mutation is associated with melanocytic neoplasms and thyroid cancer.

    Science.gov (United States)

    McDonnell, Kevin J; Gallanis, Gregory T; Heller, Kathleen A; Melas, Marilena; Idos, Gregory E; Culver, Julie O; Martin, Sue-Ellen; Peng, David H; Gruber, Stephen B

    2016-03-01

    Germline mutations in the tumor suppressor gene, BRCA-1 associated protein (BAP1), underlie a tumor predisposition syndrome characterized by increased risk for numerous cancers including uveal melanoma, melanocytic tumors and mesothelioma, among others. In the present study we report the identification of a novel germline BAP1 mutation, c.1777C>T, which produces a truncated BAP1 protein product and segregates with cancer. Family members with this mutation demonstrated a primary clinical phenotype of autosomal dominant, early-onset melanocytic neoplasms with immunohistochemistry (IHC) of these tumors demonstrating lack of BAP1 protein expression. In addition, family members harboring the BAP1 c.1777C>T germline mutation developed other neoplastic disease including thyroid cancer. IHC analysis of the thyroid cancer, as well, demonstrated loss of BAP1 protein expression. Our investigation identifies a new BAP1 mutation, further highlights the relevance of BAP1 as a clinically important tumor suppressor gene, and broadens the range of cancers associated with BAP1 inactivation. Further study will be required to understand the full scope of BAP1-associated neoplastic disease.

  7. Multidirectional Vector Excision Leads to Better Outcomes than Traditional Elliptical Excision of Facial Congenital Melanocytic Nevus

    Directory of Open Access Journals (Sweden)

    Seung Il Oh

    2013-09-01

    Full Text Available Background The elliptical excision is the standard method of removing benign skin lesions,such as congenital melanocytic nevi. This technique allows for primary closure, with little to nodog-ear deformity, but may sacrifice normal tissue adjacent to the lesion, resulting in scarswhich are unnecessarily long. This study was designed to compare the predicted results ofelliptical excision with those resulting from our excision technique.Methods Eighty-two patients with congenital melanocytic nevus on the face were prospectivelystudied. Each lesion was examined and an optimal ellipse was designed and marked onthe skin. After an incision on one side of the nevus margin, subcutaneous undermining wasperformed in the appropriate direction. The skin flap was pulled up and approximated alongseveral vectors to minimize the occurrence of dog-ear deformity.Results Overall, the final wound length was 21.1% shorter than that achieved by ellipticalexcision. Only 8.5% of the patients required dog-ear repair. There was no significant distortionof critical facial structures. All of the scars were deemed aesthetically acceptable based ontheir Patient and Observer Scar Assessment Scale scores.Conclusions When compared to elliptical excision, our technique appears to minimize dogeardeformity and decrease the final wound length. This technique should be considered analternative method for excision of facial nevi.

  8. Abundance of the benign melanocytic universe: Dermoscopic-histopathological correlation in nevi.

    Science.gov (United States)

    Woltsche, Nora; Schmid-Zalaudek, Karin; Deinlein, Teresa; Rammel, Katrin; Hofmann-Wellenhof, Rainer; Zalaudek, Iris

    2017-05-01

    The broad universe of "melanocytic nevi" includes a variety of different subtypes, which can be classified either due to their morphology, epidemiology, genetic alterations or risk for developing melanoma. Regarding morphology, on the one hand macroscopic/clinical and on the other hand histopathological appearance were used to subdivide in the past, often resulting in confusion and poor interobserver agreement, while nowadays dermoscopy presents the clinician's precious bridge between naked-eye examination and histopathological diagnostics, allowing prediction of the lesions' histopathology, follow up and monitoring over time without need of excision. The non-invasive dermoscopic examination relies on the assessment of colors, patterns and the distribution of both within a cutaneous lesion. Until today, the correspondence of certain dermoscopic colors and patterns to certain histopathological correlates has been reported for a huge amount of different cutaneous lesions. Moreover, the correspondence of certain dermoscopic features to certain body sites, age groups and pigmentary traits, but also to specific genetic alterations in lesions, has been broadly investigated. Dermoscopy has led us to a new understanding of melanocytic nevi's biology and evolution and, last but not least, to a new classification system, which we want to present herein. © 2017 Japanese Dermatological Association.

  9. 粒细胞集落刺激因子及其受体对人黑素细胞增殖及酪氨酸酶活性的影响%Effect of granulocyt e colony-stimulating factor and its receptor on the proliferation and tyrosinase activity of human melanocytes

    Institute of Scientific and Technical Information of China (English)

    周梅华; 李雪; 吴迪; 朱文元; 鲁严

    2012-01-01

    .Methods Melanocytes were obtained from circumcision specimens of healthy males,and neutrophils were isolated from heparin-andcoagulated peripheral blood of healthy human followed by a primary culture.Then,the melanocytes in third passage were cultured with or without the presence of various concentrations (200,400,600,800 μg/L) of rhG-CSF for 72 hours.The growth and morphology of melanocytes were observed.Flow cytometry was performed to detect the expression of G-CSFR in untreated human melanocytes,neutrophils and erythroleukemia cells (HEL 92.1.7).Western blot and reverse transcription PCR (RT-PCR) were carried out to measure the expression of G-CSFR protein and mRNA respectively in the neutrophils,HEL 92.1.7 cells,treated or untreated human melanocytes.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferation,and dopa-oxidation assay to estimate the tyrosinase activity,of treated melanocytes.Results The expression rate of G-CSFR was 76.81% ± 10.70% in human melanocytes,significantly higher than that in the HEL 92.1.7 cells (2.53% ± 1.54%,P < 0.01 ),but lower than that in the neutrophils (85.76% ± 15.71%,P < 0.05).Both G-CSFR protein and mRNA were expressed in melanocytes,and there was no significant differences in the expression level of G-CSFR protein and mRNA among melanocytes treated with different concentrations of rhG-CSF (both P > 0.05).The expression levels of G-CSFR protein and mRNA in the melanecytes were significantly higher than those in the HEL 92.1.7 cells (both P < 0.01 ),but lower than those in the neutrophils (P < 0.05 or < 0.01 ).rhG-CSF at 200-800 μg/L displayed a significantly promotive effect on the proliferation of melanocytes (P < 0.01 or < 0.05 ),and the effect was in a dose-dependent manner when rhG-CSF ranged from 200 to 600 μg/L (P < 0.01 ).The rhG-CSF at 600 μg/L and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) at 20 μg/L showed an equivalent effect on the proliferation of melanocytes (164

  10. 用HaCaT细胞和正常人黑素细胞构建组织工程皮肤%Reconstruction of tissue engineered skin with HaCaT cells and melanocytes in vitro

    Institute of Scientific and Technical Information of China (English)

    曹玉萍; 孙建方; 周武庆; 马鹏程; 刘维达; 李玲珺; 陶明; 桑红桂; 李阿梅; 邵雪宝

    2009-01-01

    目的 探索用人永生化表皮细胞(HacaT)和黑素细胞复合鼠尾胶原凝胶支架构建组织工程皮肤的可行性.方法 HaCaT细胞和黑素细胞与鼠尾胶原复合,经历液下和气液面共13 d的培养,苏木素-伊红(HE)染色观察不同培养阶段组织学形态变化,S-100免疫组化染色观察黑素细胞分布情况.结果 随培养时间的延长,组织工程皮肤的细胞层数逐渐增加,结束时可达12~15层;同时,细胞层内发生黑素化,颜色逐渐加深,组织学结构可见黑素颗粒分布广泛,S-100免疫组化染色显示黑素细胞均匀分布于基底层.结论 在鼠尾胶原凝胶支架表面用HaCaT细胞和黑素细胞可构建出组织工程皮肤,为探索基于组织工程皮肤的化合物光毒性检测奠定基础.%Objective To reconstruct the tissue engineered skin with HaCaT cells and melanocytes on rat tail tendon collagen gel support in vitro . Methods HaCaT cells and melanocytes were combined to rat tail tendon collagen, which then were cultured in the medium for 3 d and at the air-liquid interface for 10 d. Morphology and histology of the tissue engineered skin during different culture stages were examined with HE staining, and immune-histo-chemical staining for S-100 was conducted to analyze the distribution of melanocytes. Results As time goes by, the cell layer became thicker and thicker, and could reach 12 - 15 cell layers till the end of culture;Moreover,the tissue engineered skin was pigmented and the tanning degree was getting stronger. The melanins were distributed widely in histology and S-100 positive cells,the melanocytes,were distributed uniformly in the basal layer. Conclusion Reconstruction of the tissue engineered skin with HaCaT cells and melanocytes on rat tail tendon collagen gel support in vitro was successful, which was useful for studying the phototoxicity testing based on tissue engineered skin.

  11. Effects of ethanol extract of glossy privet fruit and its monomer tyrosol on the adhesion and migration of human epidermal melanocytes%女贞子乙醇提取物及酪醇对人表皮黑素细胞黏附和迁移的影响

    Institute of Scientific and Technical Information of China (English)

    李其林; 李湘君; 何丹华; 牛牧; 黄永华

    2013-01-01

    目的 探讨女贞子乙醇提取物及其单体酪醇对人表皮黑素细胞黏附和迁移的调节作用.方法 体外分离培养人表皮黑素细胞,XTT法测定细胞增殖情况,用经FN包被的培养板测定细胞黏附率,Transwell微孔膜法观测细胞迁移情况,激光共聚焦显微镜观察经中药处理的细胞内肌动蛋白细胞骨架的结构分布,半定量分析细胞内荧光强度.结果 0.0375~0.6 mg/ml女贞子乙醇提取物均可促进黑素细胞在FN上的黏附(P< 0.05或P< 0.01);0.5~2 mmol/L酪醇可促进黑素细胞黏附(P<0.05或P<0.01).0.15 mg/ml女贞子乙醇提取物及2 mmol/L酪醇无明显细胞毒性,在24 h时不明显促进细胞增殖,选择此浓度为工作浓度,女贞子乙醇提取物及酪醇穿过微孔滤膜的黑素细胞数分别为43.7和51.0个,显著高于对照组的20.3个(P< 0.01);两种药物工作浓度作用的黑素细胞与对照组相比,胞内可见较多呈束状的应力纤维,并集中分布于细胞膜内侧和细胞核周围,胞内荧光强度均高于对照组(P<0.01).结论 女贞子乙醇提取物及其单体酪醇在体外可促进黑素细胞的黏附及迁移,其机制可能与诱导黑素细胞内肌动蛋白细胞骨架的聚合有关.%Objective To study the regulatory effect of ethanol extract of glossy privet fruit and its monomer tyrosol on the adhesion and migration of human epidermal melanocytes.Methods Epidermal melanocytes were isolated from human foreskin,and subjected to a primary culture.Mter 3-5 passages,the melanocytes were treated with various concentrations of ethanol extract of glossy privet fruit (0.0375-0.6 mg/ml)and tyrosol (0.125-2 mmol/L) for 24-72 hours.The XTT colorimetric assay was carried out to evaluate the proliferation of melanocytes,fibronectin (FN)-coated culture plates were used to evaluate the adhesion activity of melanocytes,and Transwell assay was conducted to assess the migration activity of melanocytes.Confocal laser

  12. Retinoid-X-receptors (α/β) in melanocytes modulate innate immune responses and differentially regulate cell survival following UV irradiation.

    Science.gov (United States)

    Coleman, Daniel J; Garcia, Gloria; Hyter, Stephen; Jang, Hyo Sang; Chagani, Sharmeen; Liang, Xiaobo; Larue, Lionel; Ganguli-Indra, Gitali; Indra, Arup K

    2014-05-01

    Understanding the molecular mechanisms of ultraviolet (UV) induced melanoma formation is becoming crucial with more reported cases each year. Expression of type II nuclear receptor Retinoid-X-Receptor α (RXRα) is lost during melanoma progression in humans. Here, we observed that in mice with melanocyte-specific ablation of RXRα and RXRβ, melanocytes attract fewer IFN-γ secreting immune cells than in wild-type mice following acute UVR exposure, via altered expression of several chemoattractive and chemorepulsive chemokines/cytokines. Reduced IFN-γ in the microenvironment alters UVR-induced apoptosis, and due to this, the survival of surrounding dermal fibroblasts is significantly decreased in mice lacking RXRα/β. Interestingly, post-UVR survival of the melanocytes themselves is enhanced in the absence of RXRα/β. Loss of RXRs α/β specifically in the melanocytes results in an endogenous shift in homeostasis of pro- and anti-apoptotic genes in these cells and enhances their survival compared to the wild type melanocytes. Therefore, RXRs modulate post-UVR survival of dermal fibroblasts in a "non-cell autonomous" manner, underscoring their role in immune surveillance, while independently mediating post-UVR melanocyte survival in a "cell autonomous" manner. Our results emphasize a novel immunomodulatory role of melanocytes in controlling survival of neighboring cell types besides controlling their own, and identifies RXRs as potential targets for therapy against UV induced melanoma.

  13. Additive effect of heat on the UVB-induced tyrosinase activation and melanogenesis via ERK/p38/MITF pathway in human epidermal melanocytes.

    Science.gov (United States)

    Gu, Wei-Jie; Ma, Hui-Jun; Zhao, Guang; Yuan, Xiao-Ying; Zhang, Ping; Liu, Wen; Ma, Li-Juan; Lei, Xiao-Bing

    2014-08-01

    Heat is known as an environmental factor that causes significant skin pigmentation, but its effects on melanogenesis have been poorly studied. It has been shown that mitogen-activated protein kinase (MAPK) is involved in ultraviolet B (UVB) and stress-induced melanogenesis in melanocytes. In this study, we investigated the effects of heat and UVB, on melanocyte melanogenesis, differentiation, and MAPK phosphorylation. The results showed that heat (1 h at 40 °C for 5 days) increased cell dendrites, enlarged cell bodies, and induced extracellular signal-regulated kinases (ERK)/p38/MITF activation but did not influence melanogenesis of human epidermal melanocytes from skin phototype III. UVB irradiation (20 mJ/cm(2) for 5 days) induced melanogenesis and c-jun N-terminal kinases (JNK)/p38/MITF/tyrosinase activation in melanocytes from skin phototype III. UVB combined with heat resulted in much more significant tyrosinase activation and melanogenesis as compared with UVB alone in melanocytes from skin phototype III. Furthermore, heat treatment and UVB irradiation induced JNK, ERK, and p38 activation but not melanogenic and morphological changes in melanocytes from skin phototype I. These findings suggested that heat promoted melanocyte differentiation, probably via heat-induced ERK/p38/MITF/activation. Furthermore, heat had an additive effect on the UVB-induced tyrosinase activation and melanogenesis. These results provide a new clue for dermatologists for the treatment of hypopigmented skin disease with heat combined with UVB irradiation.

  14. Differentiating intratumoral melanocytes from Langerhans cells in nonmelanocytic pigmented skin tumors in vivo by label-free third-harmonic generation microscopy

    Science.gov (United States)

    Weng, Wei-Hung; Liao, Yi-Hua; Tsai, Ming-Rung; Wei, Ming-Liang; Huang, Hsin-Yi; Sun, Chi-Kuang

    2016-07-01

    Morphology and distribution of melanocytes are critical imaging information for the diagnosis of melanocytic lesions. However, how to image intratumoral melanocytes noninvasively in pigmented skin tumors is seldom investigated. Third-harmonic generation (THG) is shown to be enhanced by melanin, whereas high accuracy has been demonstrated using THG microscopy for in vivo differential diagnosis of nonmelanocytic pigmented skin tumors. It is thus desirable to investigate if label-free THG microscopy was capable to in vivo identify intratumoral melanocytes. In this study, histopathological correlations of label-free THG images with the immunohistochemical images stained with human melanoma black (HMB)-45 and cluster of differentiation 1a (CD1a) were made. The correlation results indicated that the intratumoral THG-bright dendritic-cell-like signals were endogenously derived from melanocytes rather than Langerhans cells (LCs). The consistency between THG-bright dendritic-cell-like signals and HMB-45 melanocyte staining showed a kappa coefficient of 0.807, 84.6% sensitivity, and 95% specificity. In contrast, a kappa coefficient of -0.37, 21.7% sensitivity, and 30% specificity were noted between the THG-bright dendritic-cell-like signals and CD1a staining for LCs. Our study indicates the capability of noninvasive label-free THG microscopy to differentiate intratumoral melanocytes from LCs, which is not feasible in previous in vivo label-free clinical-imaging modalities.

  15. SCF/c-kit signaling is required in 12-O-tetradecanoylphorbol-13-acetate-induced migration and differentiation of hair follicle melanocytes for epidermal pigmentation.

    Science.gov (United States)

    Qiu, Weiming; Yang, Ke; Lei, Mingxing; Yan, Hongtao; Tang, Hui; Bai, Xiufeng; Yang, Guihong; Lian, Xiaohua; Wu, Jinjin

    2015-05-01

    Hair follicle melanocyte stem cells (McSCs) are responsible for hair pigmentation and also function as a major melanocyte reservoir for epidermal pigmentation. However, the molecular mechanism promoting McSCs for epidermal pigmentation remains elusive. 12-O-tetradecanoylphorbol-13-acetate (TPA) mimics key signaling involved in melanocyte growth, migration and differentiation. We therefore investigated the molecular basis for the contribution of hair follicle McSCs to epidermal pigmentation using the TPA induction model. We found that repetitive TPA treatment of female C57BL/6 mouse dorsal skin induced epidermal pigmentation by increasing the number of epidermal melanocytes. Particularly, TPA treatment induced McSCs to initiate proliferation, exit the stem cell niche and differentiate. We also demonstrated that TPA promotes melanoblast migration and differentiation in vitro. At the molecular level, TPA treatment induced robust expression of stem cell factor (SCF) in keratinocytes and c-kit in melanoblasts and melanocytes. Administration of ACK2, a neutralizing antibody against the Kit receptor, suppressed mouse epidermal pigmentation, decreased the number of epidermal melanocytes, and inhibited melanoblast migration. Taken together, our data demonstrate that TPA promotes the expansion, migration and differentiation of hair follicle McSCs for mouse epidermal pigmentation. SCF/c-kit signaling was required for TPA-induced migration and differentiation of hair follicle melanocytes. Our findings may provide an excellent model to investigate the signaling mechanisms regulating epidermal pigmentation from mouse hair follicle McSCs, and a potential therapeutic option for skin pigmentation disorders.

  16. Retinoid-X-receptors (α/β in melanocytes modulate innate immune responses and differentially regulate cell survival following UV irradiation.

    Directory of Open Access Journals (Sweden)

    Daniel J Coleman

    2014-05-01

    Full Text Available Understanding the molecular mechanisms of ultraviolet (UV induced melanoma formation is becoming crucial with more reported cases each year. Expression of type II nuclear receptor Retinoid-X-Receptor α (RXRα is lost during melanoma progression in humans. Here, we observed that in mice with melanocyte-specific ablation of RXRα and RXRβ, melanocytes attract fewer IFN-γ secreting immune cells than in wild-type mice following acute UVR exposure, via altered expression of several chemoattractive and chemorepulsive chemokines/cytokines. Reduced IFN-γ in the microenvironment alters UVR-induced apoptosis, and due to this, the survival of surrounding dermal fibroblasts is significantly decreased in mice lacking RXRα/β. Interestingly, post-UVR survival of the melanocytes themselves is enhanced in the absence of RXRα/β. Loss of RXRs α/β specifically in the melanocytes results in an endogenous shift in homeostasis of pro- and anti-apoptotic genes in these cells and enhances their survival compared to the wild type melanocytes. Therefore, RXRs modulate post-UVR survival of dermal fibroblasts in a "non-cell autonomous" manner, underscoring their role in immune surveillance, while independently mediating post-UVR melanocyte survival in a "cell autonomous" manner. Our results emphasize a novel immunomodulatory role of melanocytes in controlling survival of neighboring cell types besides controlling their own, and identifies RXRs as potential targets for therapy against UV induced melanoma.

  17. Differentially expressed genes during human cutaneous melanocytic tumor progression : a focus on cancer/testis-associated genes

    NARCIS (Netherlands)

    Zendman, Albert Johan Willem

    2003-01-01

    Human cutaneous melanoma, the skin cancer originating from the pigment producing melanocyte, is one of the most aggressive types of tumors due to its early dissemination. The progression of melanoma surpasses several stages from common nevi to metastatic tumors. For diagnostic and clinical purposes

  18. Skeletal muscle-melanocyte association during tadpole tail resorption in a tropical frog, Clinotarsus curtipes Jerdon (Anura, Ranoidea).

    Science.gov (United States)

    Divya, Lekha; Beyo, Reston S; Sreejith, Parameswaran; Akbarsha, Mohammad A; Oommen, Oommen V

    2010-05-01

    We tested the hypothesis that melanin has a role as a molecule within the thyroid-mediated cascade. Light microscopic and ultrastructural changes in the skeletal muscle during tail resorption in tadpoles of the tropical frog Clinotarsus curtipes Jerdon (Anura: Ranoidea) were o