Mutations in <em>GRHL2</em> result in an autosomal-recessive ectodermal dysplasia syndrome

Grainyhead-like 2, encoded by GRHL2, is a member of a highly conserved family of transcription factors that play essential roles during epithelial development. Haploinsufficiency for GRHL2 has been implicated in autosomal-dominant deafness, but mutations have not yet been associated with any skin pathology. We investigated two unrelated Kuwaiti families in which a total of six individuals have had lifelong ectodermal defects. The clinical features comprised nail dystrophy or nail loss, marginal palmoplantar keratoderma, hypodontia, enamel hypoplasia, oral hyperpigmentation, and dysphagia. In addition, three individuals had sensorineural deafness, and three had bronchial asthma. Taken together, the features were consistent with an unusual autosomal-recessive ectodermal dysplasia syndrome. Because of consanguinity in both families, we used whole-exome sequencing to search for novel homozygous DNA variants and found GRHL2 mutations common to both families: affected subjects in one family were homozygous for c.1192T>C (p.Tyr398His) in exon 9, and subjects in the other family were homozygous for c.1445T>A (p.Ile482Lys) in exon 11. Immortalized keratinocytes (p.Ile482Lys) showed altered cell morphology, impaired tight junctions, adhesion defects, and cytoplasmic translocation of GRHL2. Whole-skin transcriptomic analysis (p.Ile482Lys) disclosed changes in genes implicated in networks of cell-cell and cell-matrix adhesion. Our clinical findings of an autosomal-recessive ectodermal dysplasia syndrome provide insight into the role of GRHL2 in skin development, homeostasis, and human disease

Analysis of Connectivity Between Bluetooth Devices and Vehicle Infotainment Systems

Cílem této práce je analyzovat spolehlivost komunikace mezi dvěma zařízeními, vybavenými bezdrátovou technologií Bluetooth. Konkrétně jsem se zaměřil na komunikaci mezi infotainmentem vozidla a mobilním telefonem. Zvolený problém jsem vyřešil pomocí opakovaného měření, které simulovalo různé rušivé vlivy na Bluetooth komunikaci. Data jsem získával pomocí Bluetooth protokolového analyzátoru a příslušného softwaru. Provedeným výzkumem jsem zjistil, že spolehlivost Bluetooth komunikace je uvnitř vozu na vysoké úrovni a největší překážkou pro Bluetooth signál je v automobilu lidské tělo.The purpose of this thesis is to analyze the reliability of communication between two devicesequipped with Bluetooth wireless technology. Specifically, I focused on communication betweena vehicle's infotainment system and a cellular phone. I have solved the defined problem bycarrying out multiple measurements which simulated various interference influences on Bluetoothcommunication. I obtained the data using a Bluetooth protocol analyzer and respective software.By undertaking this research, I have found that the reliability of Bluetooth communication insidethe vehicle is at a high grade and that the human body is the major obstacle for Bluetooth signalin the vehicle

Loss of grainy head-like 1 is associated with disruption of the epidermal barrier and squamous cell carcinoma of the skin

The Grainyhead-like 1 (GRHL1) transcription factor regulates the expression of desmosomal cadherin desmoglein 1 (Dsg1) in suprabasal layers of the epidermis. As a consequence, the epidermis of Grhl1-null mice displays fewer desmosomes that are abnormal in structure. These mice also exhibit mild chronic skin barrier defects as evidenced by altered keratinocyte terminal differentiation, increased expression of inflammatory markers and infiltration of the skin by immune cells. Exposure of Grhl1 (−/−) mice to a standard chemical skin carcinogenesis protocol results in development of fewer papillomas than in wild type control animals, but with a rate of conversion to squamous cell carcinoma (SCC) that is strikingly higher than in normal littermates. The underlying molecular mechanism differs from mice with conditional ablation of a closely related Grhl family member, Grhl3, in the skin, which develop SCC due to the loss of expression of phosphatase and tensin homolog (PTEN) and activation of the phosphatidylinositol 3-kinase (PI3K)/AKT/mechanistic target of rapamycin (mTOR) signaling pathway

Skin inflammation in <i>Grhl1</i>^−/− mice.

<p>(A) Blood levels of TSLP in <i>Grhl1</i>^+/+ (gray bar) and <i>Grhl1</i>^−/− mice (blue bar), measured with ELISA kit. (B) Levels of expression of antimicrobial peptides S100A8 and S100A9 in the epidermis of <i>Grhl1</i>^+/+ (gray bar) and <i>Grhl1</i>^−/− mice (blue bar), measured with Q-RT-PCR. (C) Representative skin sections of <i>Grhl1</i>^+/+ (top panel) and <i>Grhl1</i>^−/− mice (bottom panel) stained with toluidine blue to visualize dermal mast cells (purple cells). Scale bars represent 200 µm. (D) Quantification of skin infiltration with mast cells for <i>Grhl1</i>^+/+ (gray bar) and <i>Grhl1</i>^−/− mice (blue bar), estimated as numbers of stained cells per 1 mm² area of 10 µm thick skin section (using ImageJ software). (A, B, D) Significance (Student’s t-test, p-value) is shown above bars.</p

Histological analysis of the skin of <i>Grhl1</i>^−/− mice (right panels) in comparison to wild type littermates <i>Grhl1</i>^+/+ (left panels).

<p>Scale bars represent 25 µm. (A–B) Hematoxylin and eosin (H&E) staining of epidermis of <i>Grhl1</i>^+/+ and <i>Grhl1</i>^−/− mice; red arrows point to keratinocytes from granular layer of epidermis. (C–L) Immunohistochemical analysis of markers of epidermal differentiation in the epidermis of <i>Grhl1</i>^+/+ and <i>Grhl1</i>^−/− mice: marker of basal layer – keratin 5 (C, D, black arrows indicate suprabasal keratinocytes expressing keratin 5), marker of early terminal differentiation – keratin 10 (E, F, red arrows point to keratinocytes from the granular layer of epidermis), markers of late terminal differentiation – involucrin (G, H), filaggrin (I, J), loricrin (K, L). (M, N) Immunofluorescence staining of proliferation marker Ki67 in <i>Grhl1</i>^+/+ (M) and <i>Grhl1</i>^−/− (N) mice.</p

List of some of the genes that are upregulated in the <i>Grhl1</i>^−/− skin.

<p>Full list of genes with fold changes higher than 2 is provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0089247#pone.0089247.s003" target="_blank">Table S2</a>.</p

Pharmacological Inhibition of Chitotriosidase (CHIT1) as a Novel Therapeutic Approach for Sarcoidosis

Introduction: Sarcoidosis is a systemic disease of unknown etiology characterized by granuloma formation in the affected tissues. The pathologically activated macrophages are causatively implicated in disease pathogenesis and play important role in granuloma formation. Chitotriosidase (CHIT1), macrophage-derived protein, is upregulated in sarcoidosis and its levels correlate with disease severity implicating CHIT1 in pathology. Methods: CHIT1 was evaluated in serum and bronchial mucosa and mediastinal lymph nodes specimens from sarcoidosis patients. The therapeutic efficacy of OATD-01 was assessed ex vivo on human bronchoalveolar lavage fluid (BALF) macrophages and in vivo in the murine models of granulomatous inflammation. Results: CHIT1 activity was significantly upregulated in serum from sarcoidosis patients. CHIT1 expression was restricted to granulomas and localized in macrophages. Ex vivo OATD-01 inhibited pro-inflammatory mediators’ production (CCL4, IL-15) by lung macrophages. In the acute model of granulomatous inflammation in mice, OATD-01 showed anti-inflammatory effects reducing the percentage of neutrophils and CCL4 concentration in BALF. In the chronic model, inhibition of CHIT1 led to a decrease in the number of organized lung granulomas and the expression of sarcoidosis-associated genes. Conclusion: In summary, CHIT1 activity was increased in sarcoidosis patients and OATD-01, a first-in-class CHIT1 inhibitor, demonstrated efficacy in murine models of granulomatous inflammation providing a proof-of-concept for its clinical evaluation in sarcoidosis