Dual-specificity phosphatases in the hypo-osmotic stress response of keratin-defective epithelial cell lines

Although mutations in intermediate filament proteins cause many human disorders, the detailed pathogenic mechanisms and the way these mutations affect cell metabolism are unclear. In this study, selected keratin mutations were analysed for their effect on the epidermal stress response. Expression profiles of two keratin-mutant cell lines from epidermolysis bullosa simplex patients (one severe and one mild) were compared to a control keratinocyte line before and after challenge with hypo-osmotic shock, a common physiological stress that transiently distorts cell shape. Fewer changes in gene expression were found in cells with the severely disruptive mutation (55 genes altered) than with the mild mutation (174 genes) or the wild type cells (261 genes) possibly due to stress response pre-activation in these cells. We identified 16 immediate-early genes contributing to a general cell response to hypo-osmotic shock, and 20 genes with an altered expression pattern in the mutant keratin lines only. A number of dual-specificity phosphatases (MKP-1, MKP-2, MKP-3, MKP-5 and hVH3) are differentially regulated in these cells, and their downstream targets p-ERK and p-p38 are significantly up-regulated in the mutant keratin lines. Our findings strengthen the case for the expression of mutant keratin proteins inducing physiological stress, and this intrinsic stress may affect the cell responses to secondary stresses in patients' skin.     

Fungal community structure under goat willows (Salix caprea L.) growing at metal polluted site: the potential of screening in a model phytostabilisation study

Goat willow (Salix caprea L.) was selected in a previous vegetation screening study as a potential candidate for the later-stage phytostabilisation efforts at a heavily metal polluted site in Slovenia. The aims of this study were to identify the fungi colonising roots of S. caprea along the gradient of vegetation succession and to estimate their colonisation levels in relation to metal pollution in order to reveal its mycorrhizal status at the site. Additionally the metal accumulation capacity of S. caprea and photosynthetic pigments were analysed as indications of its fitness at four differentially polluted plots. Despite high concentrations of leaf accumulated Cd, no significant differences in photosynthetic pigment concentrations were observed. The roots were colonised by arbuscular mycorrhizal (AM) fungi, ectomycorrhizal (EM) fungi, and dark septate endophytes (DSE), with EM as the dominant type on all the plots. Molecular characterisation showed poor correlation of the root EM community with the above-ground sporocarp diversity. Members of Sordariaceae were the most frequent colonisers with an average colonisation of 21% of all root tips, followed by Thelephoraceae with 10%. DSE colonisation increased with increasing Pb concentrations and decreasing organic matter (OM).     

Epidermal stem cells: the cradle of epidermal determination, differentiation and wound healing

The field of epidermal stem cells has dramatically advanced in the last decade, leading to a better understanding of the molecular factors, signalling pathways and cellular events that identify and characterize stem cells, thus revealing their immense potential for therapeutic use. Furthermore, multipotent epidermal stem cells present the major advantage of easy accessibility with the discovery of their specific location within the bulge of the hair follicle. This review focuses on the most recent findings on epidermal stem cells, and their potential role in initial epidermal commitment, differentiation and wound healing processes in the skin.     

Isolates of dark septate endophytes reduce metal uptake and improve physiology of Salix caprea L.

Background and aims

Dark septate endophyte (DSE) fungi are the most frequent colonists on the roots of Salix caprea in highly metal-contaminated soils. The present study was performed to obtain insights into the physiology and potential role(s) of DSE fungi for Salix caprea growing in metal-enriched sites.

Methods

Fungal isolates from S. caprea roots were identified using molecular methods, and tested for their tolerance and metal accumulation in axenic cultures. In addition, an inoculation experiment was performed to monitor the effects of the fungi on the metal uptake by the plant host. Fitness of S. caprea cuttings was assessed by photosynthetic pigment quantification and measurements of transpiration.

Results

Fungal isolates were identified as members of the Phialophora/Cadophora complex. They showed different levels of metal tolerance and maintained growth on metal-enriched media. In comparison to non-inoculated cuttings, the DSE-inoculated cuttings had lower leaf Cd content, and for isolates DB146 and DB148, also lower Zn concentrations. All DSE isolates increased the chlorophyll levels of the cuttings, with isolate DB146 also positively affecting the transpiration rate of S. caprea.

Conclusions

Our findings suggest that DSE reduced the metal uptake by the S. caprea cuttings, thus suggesting a beneficial role for S. caprea in metal-enriched soils.     

Distribution and diversity of arbuscular mycorrhizal fungi in grapevines from production vineyards along the eastern Adriatic coast

The colonisation and diversity of arbuscular mycorrhizal fungi (AMF) on roots of grapevines were investigated in production vineyards located along a 500-km-long stretch of karst along the coast of the Adriatic Sea. AMF communities on roots of grapevines were analysed using temporal temperature gel electrophoresis and sequencing of the 18S and internal transcribed spacer segments of the rDNA operon. The AMF colonisation of these grapevines roots was consistent along the whole of this east Adriatic karst region, at 64 to 82 % of fine roots. The comparison of the AMF communities on the roots of these grapevines showed that the fungal community associated with grapevine roots seems to be relatively stable, with inter-vineyard variability comparable to intra-vineyard variability. Some of the changes in the fungal communities were attributed to environmental factors (plant-available P) and location of the vineyard, although the latter could also have been influenced by an unmeasured environmental factor. A total of 27 taxa of fungi were identified, including taxa from Glomus group B, based on the sequencing of 18S rDNA. Sequencing of the internal transcribed spacer rDNA yielded 30 different fungal taxa, which comprised eight different Glomeromycota taxa, including Glomus sinuosum and Glomus indicum. To our knowledge, this is the first report of grapevine colonisation by G. indicum.