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Abnormalities in the white matter microstructure of the attentional system have been implicated in the aetiology of attention deficit hyperactivity disorder (ADHD). Diffusion tensor imaging (DTI) is a promising magnetic resonance imaging (MRI) technology that has increasingly been used in studies of white matter microstructure in the brain. The main objective of this work was to perform an exploratory analysis of white matter tracts in a sample of children with ADHD versus typically developing children (TDC). For this purpose, 13 drug-naive children with ADHD of both genders underwent MRI using DTI acquisition methodology and tract-based spatial statistics. The results were compared to those of a sample of 14 age- and gender-matched TDC. Lower fractional anisotropy was observed in the splenium of the corpus callosum, right superior longitudinal fasciculus, bilateral retrolenticular part of the internal capsule, bilateral inferior fronto-occipital fasciculus, left external capsule and posterior thalamic radiation (including right optic radiation). We conclude that white matter tracts in attentional and motor control systems exhibited signs of abnormal microstructure in this sample of drug-naive children with ADHD.

During terminal differentiation of keratinocytes the expression of various proteins, which are required for the formation of the epidermal water barrier in the skin of land dwelling animals, is upregulated. Using a cell culture model for the differentiation of human keratinocytes and real-time PCR, we quantified the mRNA levels of several proteins involved in differentiation and ceramide metabolism. A calcium shift (1.1 mM CaCl2, 10 microM linoleic acid) for 8 days triggered an increase in mRNA levels of keratin 10 (75-fold), profilaggrin (55-fold), glucosylceramide synthase (40-fold), beta-glucocerebrosidase (30-fold), prosaposin (15-fold), acid sphingomyelinase (5-fold), and serine palmitoyltransferase (SPTLC2, 4-fold). However, mRNA levels of keratin 14 and acid ceramidase did not change significantly. On the other hand nitric oxide added at concentrations lower than 0.25mM stimulates proliferation of keratinocytes (Krischel et al., J. Invest. Dermatol. 111, 286-291, 1998). Accordingly, the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP, 0.2 mM) had no effect on the morphology of cultured keratinocytes, whereas in cultured human fibroblasts apoptosis was induced. The expression patterns obtained suggest that keratinocytes remain in a basal proliferative state, with a 3-fold increase in keratin 14 expression, a marked decrease in mRNA levels of differentiation markers and of most ceramide-metabolizing enzymes to negligible levels. The inhibitor of the NO synthase, N(G)-nitro-L-arginine-methyl ester (L-NAME, 10 mM), induced a transient increase in ceramide formation, followed by apoptosis in keratinocytes but not in fibroblasts. Both, SNAP and L-NAME, decreased the mRNA levels of all proteins involved in ceramide metabolism.

Epidermal keratinocytes are exposed to a low water concentration at the stratum corneum-stratum granulosum interface. When epithelial tissues are osmotically perturbed, cellular protection and cell volume regulation is mediated by accumulation of organic osmolytes such as taurine. Previous studies reported the presence of taurine in the epidermis of several animal species. Therefore, we analyzed human skin for the presence of the taurine transporter (TAUT) and studied the accumulation of taurine as one potential mechanism protecting epidermal keratinocytes from dehydration. According to our results, TAUT is expressed as a 69 kDa protein in human epidermis but not in the dermis. For the epidermis a gradient was evident with maximal levels of TAUT in the outermost granular keratinocyte layer and lower levels in the stratum spinosum. No TAUT was found in the basal layer or in the stratum corneum. Keratinocyte accumulation of taurine was induced by experimental induction of skin dryness via application of silica gel to human skin. Cultured human keratinocytes accumulated taurine in a concentration- and osmolarity-dependent manner. TAUT mRNA levels were increased after exposure of human keratinocytes to hyperosmotic culture medium, indicating osmosensitive TAUT mRNA expression as part of the adaptation of keratinocytes to hyperosmotic stress. Keratinocyte uptake of taurine was inhibited by beta-alanine but not by other osmolytes such as betaine, inositol, or sorbitol. Accumulation of taurine protected cultured human keratinocytes from both osmotically induced and ultraviolet-induced apoptosis. Our data indicate that taurine is an important epidermal osmolyte required to maintain keratinocyte hydration in a dry environment.

Butyltins are widely used biocides and accumulate in the food chain. Tributyltin is an imposex-inducing endocrine disrupter in animals. Imposex is characterized by the development of additional male sex organs on females. In a previous study, we identified tributyltin as an inhibitor of human cytochrom P450 aromatase activity. The present work focuses on the impact of butyltins on human androgen metabolism. Activation of androgens is mediated by two human 5alpha-reductase isoenzymes. 5alpha-Reductase type 1 was completely inhibited by tributyltin chloride (IC50=19.9 microM) and dibutyltin dichloride (IC50=32.9 microM), whereas 5alpha-reductase type 2 was only inhibited by tributyltin chloride (IC50=10.8 microM). Both isoenzymes were not affected by tetrabutyltin or monobutyltin indicating that at least two butyl groups bound to the positively charged Sn are required for the interaction of butyltins with the enzymes. Tributyltin inhibited 5alpha-reductase type 1 competitively whereas an irreversible inhibition was evident for the type 2 isoenzyme. In contrast to the distinct effects on 5alpha-reductases, reductive brain 17beta-hydroxysteroid dehydrogenase activity was not inhibited by any butyltin. Insufficient activation of androgens is responsible for developmental disorders of the male reproductive system such as hypospadias. At pharmacologic levels butyltins might contribute to the onset of developmental disorders of the male reproductive system. At present, however, it is unknown whether these levels are reached after acute or chronic exposure to butyltins.

Epidermal differentiation results in the formation of the extracellular lipid barrier in the stratum corneum, which mainly consists of ceramides, free fatty acids, and cholesterol. Differentiating keratinocytes of the stratum granulosum synthesize a series of complex long-chain ceramides and glucosylceramides with different chain lengths and hydroxylation patterns at intracellular membranes of the secretory pathway. Formation of complex extracellular ceramides parallels the transition of keratinocytes from the stratum granulosum to the stratum corneum, where their precursors, complex glucosylceramides and sphingomyelin, are secreted and exposed to extracellular lysosomal lipid hydrolases. Submerged cultures used so far showed a reduced ceramide content compared to the native epidermis or the air-exposed, organotypic culture system. In order to investigate the sphingolipid metabolism during keratinocyte differentiation, we optimized a simple cell culture system to generate the major barrier sphingolipids. This optimized model is based on the chemically well-defined serum-free MCDB medium. At low calcium ion concentrations (0.1mM), keratinocytes proliferate and synthesize mainly Cer(NS) and a small amount of Cer(NP). Supplementation of the MCDB cell culture medium with calcium ions (1.1mM) and 10 microM linoleic acid triggered differentiation of keratinocytes and synthesis of a complex pattern of free and covalently bound ceramides as found in native epidermis or air-exposed organotypic cultures, though at a reduced level. The mRNA levels of the differentiation markers keratin 10 and profilaggrin increased, as well as those of ceramide glucosyltransferase and glucosylceramide-beta-glucosidase. The described culture system was thus suitable for biochemical studies of the sphingolipid metabolism during keratinocyte differentiation. The addition of serum or vitamin A to the medium resulted in a decrease in ceramide and glucosylceramide content. Lowering the medium pH to 6, while maintained cell viability, led to an increase in the processing of probarrier lipids glucosylceramide and sphingomyelin to free ceramides and protein-bound ceramide Cer(OS).

Epidermal barrier function to water loss is maintained by lipid membrane domains located in the interstices of the stratum corneum. Exposure of the epidermis to a dry environment or UV irradiation stimulates barrier lipid synthesis and accumulation of the organic osmolyte taurine in the outermost granular keratinocyte layer. In this work we studied a possible relationship between these two different epidermal responses to environmental challenges. As a model system we selected anionic surfactant-induced barrier perturbation. Incubation of reconstructed epidermis with taurine inhibited cytotoxic and proinflammatory effects induced by sodium dodecyl sulfate including (i) a decrease in interleukin-1 alpha and prostaglandin E2 release, (ii) stabilization of keratinocyte membrane integrity, and (iii) improvement of keratinocyte viability. Repeated exposure of human skin to sodium dodecyl sulfate induced an increase in transepidermal water loss, inflammation, and hyperplasia. Topical application of taurine significantly decreased transepidermal water loss after repeated exposure to sodium dodecyl sulfate. Moreover, taurine significantly stimulated the synthesis of all three classes of barrier lipids (ceramides, cholesterol, and fatty acids) in reconstructed epidermis. In conclusion, our data suggest a role for taurine in preventing surfactant-induced dry and scaly skin by modulating the proinflammatory response and stimulating epidermal lipid synthesis.

Recent studies have provided new insights into the occurrence, causes, and pathogenetic consequences of changes in the skin pH in atopic dermatitis, particularly with respect to skin barrier function and colonization with Staphylococcus aureus. Growing evidence suggests an impaired release of proton donors, such as amino acids, urocanic acid, and lactic acid, to the stratum corneum in atopic dermatitis, as a result of reductions in filaggrin proteolysis and sweat secretion. In addition, an impaired formation of free fatty acids from sebaceous lipids and epidermal phospholipids seems to be involved. Because both lipid organization and lipid metabolism in the stratum corneum requires an acidic pH, these alterations might contribute to the disturbance of skin barrier function observed in atopic dermatitis. Furthermore, bacterial growth and virulence of S. aureus, as well as defensive host mechanisms, have increasingly been delineated as pH dependent, giving rise to a new understanding of the pathophysiology underlying increased skin colonization seen in atopic dermatitis.

Lead concentrations and lead isotope ratios were analyzed in two firn/ice cores covering the period from 1650 to 1994, which were obtained from the 4450 m high glacier saddle Colle Gnifetti located in the Monte Rosa massif at the Swiss-Italian border. This study presents the first glaciochemical time series with annual resolution, spanning several centuries of lead concentrations and lead isotopic compositions in precipitation in Europe. Lead concentrations in firn dated from the 1970s are approximately 25 times higher than in ice dated from the 17th century, confirming the massive rise in lead pollution in Europe during the last few centuries. A decline of the lead concentration is then observed during the last two decades, i.e., from 1975 to 1994. The lead isotope ratio 206Pb/207Pb decreased from about 1.18 in the 17th and 18th centuries to about 1.12 in the 1970s. These variations are in good agreement with available information on variations in anthropogenic lead emissions from West European countries, especially from the use of lead additives in gasoline.