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  • 11
    Electronic Resource
    Electronic Resource
    Springer
    Archives of dermatological research 281 (1989), S. 40-44 
    ISSN: 1432-069X
    Keywords: Hermansky-Pudlak syndrome ; Calcium ; Free radicals ; Keratinocyte cell cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cell cultures of keratinocytes, established from four Hermansky-Pudlak, syndrome (HPS) homozygotes yielded low membrane-associated thioredoxin reductase activities compared with normal healthy adult controls. This low activity has been shown to be caused by a special sensitivity of the enzyme to calcium. 45Calcium has been used to compare the kinetics for the transport and bioaccumulation of this regulatory cation in keratinocyte cultures of a kindred with HPS (i.e., one HPS homozygote, one HPS obligate heterozygote, one normal family member, and healthy adult controls). The results show that both HPS-homozygous and-heterozygous cells bind more extracellular calcium than noncarriers of this genetic defect, and HPS homozygous cells appear to have a defective calcium transport system.
    Type of Medium: Electronic Resource
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  • 12
    Electronic Resource
    Electronic Resource
    Springer
    Archives of dermatological research 282 (1990), S. 168-171 
    ISSN: 1432-069X
    Keywords: Azelaic acid ; Inhibitor ; Tyrosinase ; Thioredoxin reductase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Azelaic acid, and other saturated dicarboxylic acids (C9-C12), are shown to be competitive inhibitors of tyrosinase (K I azelaic acid = 2.73×10−3 M) and of membrane-associated thioredoxin reductase (K I azelaic acid = 1.25×10−5 M). The monomethyl ester of azelaic acid does not inhibit thioredoxin reductase, but it does inhibit tyrosinase, although double the concentration is necessary compared with azelaic acid (K I azelaic acid monomethyl ester = 5.24×10−3 M). Neither azelaic acid nor its monomethyl ester inhibit tyrosinase when catechol is used as a substrate instead of l-tyrosine. Therefore, the weak inhibitory action of azelaic acid on tyrosinase appears to be due to the competition of a single carboxylate group on this inhibitor for the α-carboxylate binding site of the l-tyrosine substrate on the enzyme active site. Based on the inhibitor constant on tyrosinase, at least cytotoxic levels of azelaic acid would be required for the direct inhibition of melanin biosynthesis in melanosomes if this mechanism is responsible for depigmentation in the hyperpigmentation disorders lentigo maligna and melasma. Alternatively only 10−5 M azelaic acid is required to inhibit thioredoxin reductase. This enzyme is shown to regulate tyrosinase through a feedback mechanism involving electron transfer to intra-cellular thioredoxin, followed by a specific interaction between reduced thioredoxin and tyrosinase. Furthermore, the thioredoxin reductase/thioredoxin system is shown to be a principal electron donor for the ribonucleotide reductases which regulates DNA synthesis. Inhibition of thioredoxin reductase by azelaic acid provides a rationale for both its depigmenting property and the reversible inhibition of DNA synthesis observed in cultured epidermal cells and also in some of the bacteria associated with acne vulgaris.
    Type of Medium: Electronic Resource
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  • 13
    Electronic Resource
    Electronic Resource
    Springer
    Archives of dermatological research 282 (1990), S. 526-531 
    ISSN: 1432-069X
    Keywords: UV radiation ; Thioredoxin reductase ; Sun blockers ; Free radical defence ; Pigmentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Defence against oxidative damage by UV-generated free radicals in both guinea pig and human skin has been found to be mediated by the ubiquitous thioprotein, thioredoxin reductase. Human keratinocytes contain approximately 5% thioredoxin reductase in their total acidic protein fraction and also express membrane-associated enzyme activity in cells cultured in synthetic medium. The thioredoxin reductase/thioredoxin system has been shown to reduce superoxide anion radicals through hydrogen peroxide to water. However, both UVA and UVB radiation, below the minimal erythemal dose, generate a sufficiently high concentration of oxygen radicals to deactivate thioredoxin reductase considerably. In albino guinea pigs, enzyme deactivation was up to 70% for UVA and 66% for UVB (n=10 animals/protocol). The application of sun blockers SPF4, SPF8 and SPF15 to albino guinea pig skin offered no significant protection for the deactivation of thioredoxin reductase by either UVA or UVB radiation. In the human population (n=15), thioredoxin reductase was deactivated by 54% with UVA and 36% with UVB radiation, although the degree of enzyme inhibition depended on skin phototype (I–VI, Fitzpatrick Classification). SPF24 offered considerable protection for thioredoxin reductase against both UVA and UVB for skin types I and II. However, SPF24 yielded no significant protection with UVA for skin types III–VI, and enhanced the enzyme inhibition with UVB additively. These results indicate that UVB photo-oxidation of oxybenzone (the UVA filter in SPF24) may deactivate thioredoxin reductase in more pigmented members of the population by Michael addition of oxybenzone semiquinone to the thiolate active site of this enzyme.
    Type of Medium: Electronic Resource
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  • 14
    ISSN: 1432-069X
    Keywords: Key words Catecholamines ; MAO-A ; Atopic eczema ; Phenylethanolamine-N-methyl-transferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Patients with atopic eczema have significantly higher norepinephrine levels in plasma than healthy controls. In addition, significantly higher levels of the essential cofactor (6R)-l-erythro-5,6,7,8-tetrahydrobiopterin (6BH4) were found in this patient group. Cell extracts from epidermal suction blister roofs revealed only half the normal activity of phenylethanolamine-N-methyl transferase (PNMT) together with a threefold induction of the norepinephrine-degrading enzyme monoamine oxidase A (MAO-A). Taken together, these results support earlier observations of a defective catecholamine/adrenoceptor signal in patients with atopic eczema.
    Type of Medium: Electronic Resource
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  • 15
    ISSN: 1432-069X
    Keywords: Beta2-adrenoceptors ; Calcium ; Vitiligo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Keratinocytes were established in serum-free culture medium from lesional and nonlesional skin of a patient with vitiligo (skin type III) and from an age-matched healthy control subject. Both differentiating and undifferentiated cells were examined for the presence of beta2-adrenoceptors in culture medium containing either low (0.1×10−3 M) or high (1.5×10−3 M) calcium concentrations. Binding experiments were performed with saturating levels of radiolabeled (—)-[3H] CGP 12177, a nonselective beta-adrenergic antagonist. Controls for nonspecific binding were determined by the addition of the beta-adrenergic antagonist, propranolol (5 Μmol), before the introduction of (—)-[3H] CGP 12177 to cell cultures. Undifferentiated keratinocytes yielded the highest expression of beta2-adrenoceptors, whereas differentiating keratinocytes grown in medium with a low calcium concentration (0.1×10−3 M) had a significantly lower expression of receptors with the exception of vitiliginous cells, which retained high densities of receptors, similar to undifferentiated cells. In addition, these vitiliginous keratinocytes showed a defect in 45calcium uptake. In contrast, differentiated keratinocytes from all three cell strains, grown in medium containing a high calcium concentration (1.5×10−3 M) revealed a significantly lower receptor density compared to undifferentiated cells. This finding identified the importance of the extracellular calcium concentration in the expression of beta2-adrenoceptors. Our results: (1) confirmed the importance of beta2-adrenoceptors in the regulation of intracellular calcium concentrations in keratinocytes; (2) showed an increase in catecholamine receptors in differentiating vitiliginous keratinocytes compared to cells established from nonlesional and control skin cultured in medium with a low calcium concentration (0.1×10−3 M); and (3) demonstrated the importance of the extracellular calcium concentration in the down-regulation of the beta2-adrenoceptors in vitiliginous keratinocytes. These new observations further suggest an association between the sympathetic nervous system and the pathogenesis of vitiligo.
    Type of Medium: Electronic Resource
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  • 16
    ISSN: 1432-069X
    Keywords: Key words Monoamine oxidase A ; Catecholamines ; Vitiligo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Human keratinocytes under in vitro conditions synthesize norepinephrine and epinephrine, whereas melanocytes lack this capacity. Keratinocytes established from lesional and nonlesional skin of patients with vitiligo synthesized four and two times more norepinephrine, respectively, than controls. Epinephrine synthesis was similar in keratinocytes from uninvolved epidermis and controls, but cells from involved skin had 6.5-fold less epinephrine than controls, indicative of low phenylehtanolamine- N -methyl transferase (PNMT) activity. Similar results were obtained in five patients with vitiligo who showed low epinephrine levels in involved epidermis. Both human keratinocytes and melanocytes expressed significant levels of monoamine oxidase A (MAO-A) activities as shown using 14 C-labelled 5-hydroxytryptamine as substrate and immunohistochemical staining with mouse monoclonal antibody. MAO-A activities in the total epidermis of patients with vitiligo were increased five- to ten-fold compared with skin of type-matched controls. Similar increases in MAO-A activities were also found in both keratinocytes and melanocytes established in vitro from vitiliginous epidermis. Based on these results, it can be concluded that defective catecholamine synthesis in the epidermis of patients with vitiligo leads to increased levels of norepinephrine with a concomitant increase in MAO-A activity.
    Type of Medium: Electronic Resource
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