Deficiency of the human cysteine protease inhibitor cystatin M/E causes hypotrichosis and dry skin

Purpose: We aimed to assess the biological and clinical significance of the human cysteine protease inhibitor cystatin M/E, encoded by the CTS6 gene, in diseases of human hair and skin. Methods: Exome and Sanger sequencing was performed to reveal the genetic cause in two related patients with hypotrichosis. Immunohistochemical, biophysical, and biochemical measurements were performed on patient skin and 3D-reconstructed skin from patient-derived keratinocytes. Results: We identified a homozygous variant c.361C>T (p.Gln121*), resulting in a premature stop codon in exon 2 of CST6 associated with hypotrichosis, eczema, blepharitis, photophobia and impaired sweating. Enzyme assays using recombinant mutant cystatin M/E protein, generated by site-directed mutagenesis, revealed that this p.Gln121* variant was unable to inhibit any of its three target proteases (legumain and cathepsins L and V). Three-dimensional protein structure prediction confirmed the disturbance of the protease/inhibitor binding sites of legumain and cathepsins L and V in the p.Gln121* variant. Conclusion: The herein characterized autosomal recessive hypotrichosis syndrome indicates an important role of human cystatin M/E in epidermal homeostasis and hair follicle morphogenesis

Proliferation, differentiation, and inflammation in normal and hyperproliferative skin using multiparameter flow cytometry.

Item does not contain fulltextOur knowledge of the etiology and pathogenesis of skin diseases characterized by abnormal growth is relatively limited. Even more important, still very little is known of how epidermopoeisis is controlled in normal epidermis. There is no cure for skin diseases caused by abnormal growth control, such as psoriasis. Mechanisms are complex, additional models for epidermal growth and differentiation, and specific techniques to analyze these processes, are needed. Therefore, we have developed flow cytometric techniques to study epidermal growth over the past two decades. A prerequisite for accurate and reliable flow cytometric analysis is a high quality of epidermal single-cell suspensions. In this chapter, protocols are described for preparation of single-cell suspensions and protocols for the multiparameter flow cytometric analysis of growth behavior in normal and hyperproliferative epidermis

Novel cytometric techniques to study proliferation and differentiation of human keratinocytes

Contains fulltext : mmubn000001_130540080.pdf (publisher's version ) (Open Access)Promotores : P. Mier, R. Happle, J. Schalkwijk en P. van de Kerkhof170 p

Transglutaminase-positive cells in psoriatic epidermis during treatment with calcitriol (1alpha,25 dihydroxy vitamin D3) and tacalcitol (1alpha,24 dihydroxy vitamin D3)

Contains fulltext : 21393___.PDF (publisher's version ) (Open Access

Epidermal cell DNA content and intermediate filaments keratin 10 and vimentin after treatment of psoriasis with calcipotriol cream once daily, twice daily and in combination with clobetasone 17-butyrate cream or betamethasone 17-valerate cream: a comparative flow cytometric study

Contains fulltext : 23825.PDF (publisher's version ) (Open Access

Demonstration of an Na+/H+ exchanger in mouse keratinocytes measured by the novel pH-sensitive fluorochrome SNARF-calcein

Contains fulltext : 25591___.PDF (publisher's version ) (Open Access

Does antisense make sense in dermatology?

The concept of antisense technology is elegant but misleadingly simple. Short oligodeoxynucleotides (ODNs) complementary to a target messenger RNA form DNA-RNA hybrids by Watson-Crick base pairing rules, and interfere with expression of the encoded protein. The potential sequence specificity of antisense ODNs makes them attractive as molecular drugs in the treatment of human diseases. The skin is readily accessible and, in theory, is therefore suitable for application of antisense ODNs. Targeted and selective inhibition of keratinocyte gene expression in human epidermis could be an efficient and safe pharmacological approach in a number of skin diseases. Based on recent studies from our group and others, in this review we present our view on the usefulness of antisense ODN technology in skin for the modulation of gene expression related to skin diseases. It has become clear from these studies that practising antisense technology requires careful experimentation and critical data interpretation. Although the antisense technique was applied with success in some skin model systems, we feel that the technology is still in its infancy. The basic questions have been answered, but there are still many more that need to be addressed

The dynamics of the response of normal skin to single and multiple epicutaneous leukotriene B4 applications analysed by three-colour flow cytometry

Contains fulltext : 21281.pdf (publisher's version ) (Open Access

Minimally-invasive Sampling of Interleukin-1alpha and Interleukin-1 Receptor Antagonist from the Skin: A Systematic Review of In vivo Studies in Humans

Contains fulltext : 177162.pdf (publisher's version ) (Open Access)Interleukin-1alpha (IL-1alpha) and its receptor antagonist IL-1RA play a pivotal role in skin homeostasis and disease. Although the use of biopsies to sample these cytokines from human skin is widely employed in dermatological practice, knowledge about less invasive, in vivo sampling methods is scarce. The aim of this study was to provide an overview of such methods by systematically reviewing studies in Medline, EMBASE, Web of Science and Cochrane Library using combinations of the terms "IL-1alpha", IL-1RA", "skin", "human", including all possible synonyms. Quality was assessed using the STrengthening the Reporting of OBservational studies in Epidemiology (STROBE) checklist. The search, performed on 14 October 2016, revealed 10 different sampling methods, with varying degrees of invasiveness and wide application spectrum, including assessment of both normal and diseased skin, from several body sites. The possibility to sample quantifiable amounts of cytokines from human skin with no or minimal discomfort holds promise for linking clinical outcomes to molecular profiles of skin inflammation

Multiparameter flow cytometry as a tool to evaluate antipsoriatic therapy

Contains fulltext : 24456.PDF (publisher's version ) (Open Access