Dysbiosis of skin microbiota with increased fungal diversity is associated with severity of disease in atopic dermatitis

Background: Atopic dermatitis (AD) is a multifactorial inflammatory skin disease and an altered skin microbiota with an increase of Staphylococcus aureus has been reported. However, the role of fungi remains poorly investigated. Objectives: We aimed to improve the understanding of the fungal skin microbiota, the mycobiota, in AD in relation to the bacterial colonization. Methods: Skin swabs of 16 AD patients and 16 healthy controls (HC) from four different skin sites, that is antecubital crease, dorsal neck, glabella and vertex from multiple time points were analysed by DNA sequencing of the internal transcribed spacer region 1 (ITS1) and 16S rRNA gene for fungi and bacteria, respectively. Results: Malassezia spp. were the predominant fungi in all subjects but with a decreased dominance in severe AD patients in favour of non-Malassezia fungi, for example Candida spp. For bacteria, a decrease of Cutibacterium spp. in AD patients in favour of Staphylococcus spp., particularly S. aureus, was observed. Further, both bacterial and fungal community compositions of severe AD patients significantly differed from mild-to-moderate AD patients and HC with the latter two having overall similar microbiota showing some distinctions in bacterial communities. Conclusions: We conclude that severe AD is associated with a pronounced dysbiosis of the microbiota with increased fungal diversity. Potentially infectious agents, for example Staphylococcus and Candida, were increased in severe AD. Keywords: atopic dermatitis; bacteria; disease severity; fungi; skin microbiot

Increased fibrosis in a mouse model of anti-laminin 332 mucous membrane pemphigoid remains unaltered by inhibition of aldehyde dehydrogenase

Mucous membrane pemphigoid (MMP) is an autoimmune blistering disease characterized by autoantibodies against the basal membrane zone of skin and surface-close epithelia and predominant mucosal lesions. The oral cavity and conjunctivae are most frequently affected, albeit clinical manifestations can also occur on the skin. MMP-associated lesions outside the oral cavity typically lead to scarring. Mechanisms underlying scarring are largely unknown in MMP and effective treatment options are limited. Herein, we assessed the collagen architecture in tissue samples of an antibody-transfer mouse model of anti-laminin-332 MMP. In MMP mice, increased collagen fibril density was observed in skin and conjunctival lesions compared to mice injected with normal rabbit IgG. The extracellular matrix of MMP skin samples also showed altered post-translational collagen cross-linking with increased levels of both lysine- and hydroxylysine-derived collagen crosslinks supporting the fibrotic phenotype in experimental MMP compared to control animals. In addition, we evaluated a potential anti-fibrotic therapy in experimental anti-laminin-332 MMP using disulfiram, an inhibitor of the aldehyde dehydrogenase (ALDH), which has been implicated in immune-mediated mucosal scarring. In addition, disulfiram also acts as a copper chelator that was shown to block lysyl oxidase activity, an enzyme involved in formation of collagen crosslinks. Topical use of disulfiram (300 μM in 2 [w/v] methocel) did not improve ocular lesions in experimental MMP over the 12-day treatment period in disulfiram-treated mice compared to vehicle-treated mice (n=8/group). Furthermore, C57BL6/J mice (n=8/group) were treated prophylactically with 200 mg/kg p.o. disulfiram or the solvent once daily over a period of 12 days. Systemic treatment did not show any reduction in the severity of oral and ocular lesions in MMP mice, albeit some improvement in skin lesions was observed in disulfiram- vs. vehicle-treated mice (p=0.052). No reduction in fibrosis was seen, as assessed by immunohistochemistry. Whilst blocking of ALDH failed to significantly ameliorate disease activity, our data provide new insight into fibrotic processes highlighting changes in the collagenous matrix and cross-linking patterns in IgG-mediated MMP

Inhibition of phosphodiesterase-4 significantly decreases oral mucosa lesions in experimental anti-laminin 332 mucous membrane pemphigoid

Mucous membrane pemphigoid (MMP) is characterized by autoantibodies against the dermal-epidermal-junction and a mucosal disease predominating over skin involvement. As the treatment of MMP patients still relies on high-dose corticosteroids, there is an unmet need for new and more specific therapies. Here, we made use of a recently established experimental model that recapitulated major clinical and immunopathological characteristics of human MMP by the injection of IgG against the murine alpha3 chain of laminin 332 (Lam332) into adult mice. In a prophylactic approach, the specific PDE4 inhibitor roflumilast (ROF) led in 2 independent, blinded experiments, to the reduction of oral lesions compared to vehicle-treated mice as quantified by endoscopy (p¼0.029). In contrast, an increase in skin lesions was observed in ROF-treated mice (p<0.0001). In a quasi-therapeutic approach, i.e. when ROF/vehicle was not used until mice had developed first skin lesions, ROF-treated mice showed significantly less oral lesions compared to vehicle-treated mice, while skin lesions did not differ. To investigate the discrepant effect of ROF on oral and skin lesions, a transcriptome analysis of both tissues in ROF- and vehicle-treated anti-Lam332 MMP mice as well as mice injected with normal rabbit IgG was performed. An up-regulation of IL-6 and an impact of CXCL2 were found by Gene Set Enrichment and STRING analysis, respectively, in both the skin and buccal mucosa of vehicle-treated mice. The subsequent incubation of murine keratinocytes with anti-mLam332 IgG resulted in a dose-dependent release of IL-6 and CXCL2, which was inhibited by the addition of ROF. Our data propose IL-6 and CXCL2 as relevant pathogenic factors in MMP and suggest PDE4 inhibition as potential novel treatment options for MMP patients with severe oral lesions

Complete genome sequence of Haloterrigena turkmenica type strain (4kT)

Haloterrigena turkmenica (Zvyagintseva and Tarasov 1987) Ventosa et al. 1999, comb. nov. is the type species of the genus Haloterrigena in the euryarchaeal family Halobacteriaceae. It is of phylogenetic interest because of the yet unclear position of the genera Haloterrigena and Natrinema within the Halobacteriaceae, which created some taxonomic problems historically. H. turkmenica, was isolated from sulfate saline soil in Turkmenistan, is a relatively fast growing, chemoorganotrophic, carotenoid-containing, extreme halophile, requiring at least 2 M NaCl for growth. Here we describe the features of this organism, together with the complete genome sequence, and annotation. This is the first complete genome sequence of the genus Haloterrigena, but the eighth genome sequence from a member of the family Halobacteriaceae. The 5,440,782 bp genome (including six plasmids) with its 5,287 protein-coding and 63 RNA genes is part of the Genomic Encyclopedia of Bacteria and Archaea project

GENIUS. Generating software-ergonomic user interfaces

GENIUS (GENerator for user Interfaces Using Software-ergonomic rules) comprises a method and the supporting tool environment for the generation of user interfaces from extended data models by means of software-ergonomic rules. The representation of the user interface is based on views defined for the data model. The basic dialogue structure is derived from the data model structure. This ensures the development of task-appropriate user interfaces by transferring the characteristics of the application domain and the user's tasks reflected in the data model to the dialogue structure. The automatic generation of the user interface from the defined views is carried out by a rule-based system with explicit design rules derived from existing guidelines. Output is generated for an existing user interface management system. The software-ergonomic rules in the generation process guarantee the consistent use of interaction objects and a uniformed dialogue structure. The use of the data model as t he starting point for the generation of the user interface ensures the integration of software engineering and user interface design by the consistent use of data for application and user interface development. The generation with GENIUS reduces the development effort and improves the quality of the user interface

Ingenieurmäßige Entwicklung von Informationssystemen. Teil 1. Grundlegende Methoden und Techniken

Bei der Softwareentwicklung ergibt sich die Notwendigkeit, durch standardisierte Abläufe nach anerkannten Regeln Softwareprodukte mit gesicherter Qualität in kurzen Durchlaufzeiten bei hoher funktionaler Wettbewerbsfähigkeit des Produkts herzustellen. In Analogie zur Entwicklung in der teilefertigenden und der montierenden Industrie wurde dafür der Begriff des Software-Engineerings geprägt. Methoden und Techniken (erster Teil), Werkzeuge (zweiter Teil) und Modelle (dritter Teil) des Software-Engineerings werden in diesem dreiteiligen Beitrag vorgestellt