Identification of Rothia Bacteria as Gluten-Degrading Natural Colonizers of the Upper Gastro-Intestinal Tract

Gluten proteins, prominent constituents of barley, wheat and rye, cause celiac disease in genetically predisposed subjects. Gluten is notoriously difficult to digest by mammalian proteolytic enzymes and the protease-resistant domains contain multiple immunogenic epitopes. The aim of this study was to identify novel sources of gluten-digesting microbial enzymes from the upper gastro-intestinal tract with the potential to neutralize gluten epitopes.Oral microorganisms with gluten-degrading capacity were obtained by a selective plating strategy using gluten agar. Microbial speciations were carried out by 16S rDNA gene sequencing. Enzyme activities were assessed using gliadin-derived enzymatic substrates, gliadins in solution, gliadin zymography, and 33-mer α-gliadin and 26-mer γ-gliadin immunogenic peptides. Fragments of the gliadin peptides were separated by RP-HPLC and structurally characterized by mass spectrometry. Strains with high activity towards gluten were typed as Rothia mucilaginosa and Rothia aeria. Gliadins (250 µg/ml) added to Rothia cell suspensions (OD(620) 1.2) were degraded by 50% after ∼30 min of incubation. Importantly, the 33-mer and 26-mer immunogenic peptides were also cleaved, primarily C-terminal to Xaa-Pro-Gln (XPQ) and Xaa-Pro-Tyr (XPY). The major gliadin-degrading enzymes produced by the Rothia strains were ∼70-75 kDa in size, and the enzyme expressed by Rothia aeria was active over a wide pH range (pH 3-10).While the human digestive enzyme system lacks the capacity to cleave immunogenic gluten, such activities are naturally present in the oral microbial enzyme repertoire. The identified bacteria may be exploited for physiologic degradation of harmful gluten peptides

A deletion and a duplication in distal 22q11.2 deletion syndrome region. Clinical implications and review

<p>Abstract</p> <p>Background</p> <p>Individuals affected with DiGeorge and Velocardiofacial syndromes present with both phenotypic diversity and variable expressivity. The most frequent clinical features include conotruncal congenital heart defects, velopharyngeal insufficiency, hypocalcemia and a characteristic craniofacial dysmorphism. The etiology in most patients is a 3 Mb recurrent deletion in region 22q11.2. However, cases of infrequent deletions and duplications with different sizes and locations have also been reported, generally with a milder, slightly different phenotype for duplications but with no clear genotype-phenotype correlation to date.</p> <p>Methods</p> <p>We present a 7 month-old male patient with surgically corrected ASD and multiple VSDs, and dysmorphic facial features not clearly suggestive of 22q11.2 deletion syndrome, and a newborn male infant with cleft lip and palate and upslanting palpebral fissures. Karyotype, FISH, MLPA, microsatellite markers segregation studies and SNP genotyping by array-CGH were performed in both patients and parents.</p> <p>Results</p> <p>Karyotype and FISH with probe N25 were normal for both patients. MLPA analysis detected a partial <it>de novo </it>1.1 Mb deletion in one patient and a novel partial familial 0.4 Mb duplication in the other. Both of these alterations were located at a distal position within the commonly deleted region in 22q11.2. These rearrangements were confirmed and accurately characterized by microsatellite marker segregation studies and SNP array genotyping.</p> <p>Conclusion</p> <p>The phenotypic diversity found for deletions and duplications supports a lack of genotype-phenotype correlation in the vicinity of the LCRC-LCRD interval of the 22q11.2 chromosomal region, whereas the high presence of duplications in normal individuals supports their role as polymorphisms. We suggest that any hypothetical correlation between the clinical phenotype and the size and location of these alterations may be masked by other genetic and/or epigenetic modifying factors.</p

Toll-Like Receptor 2, Toll-Like Receptor 4, Myeloid Differentiation Response Gene 88, and Toll-IL-1 Receptor Domain-Containing Adaptor-Inducing Interferon-gamma (TRIF) Selectively Regulate Susceptibility of PO106-125-Induced Murine Experimental Autoimmune Neuritis

The functional relevance of the innate immune system has not yet been dissected in PO106-125-induced murine experimental autoimmune neuritis. Therefore, the role of Toll-like receptor (TLR) 2, TLR4, myeloid differentiation response gene 88, and ToLL IL-1 receptor domain-containing adaptor-inducing interferon-i (TRIF), factors critically involved in the TLR signaling pathway, was studied in experimental autoimmune neuritis. In the absence of TLR2, TLR4, myeloid differentiation response gene 88, or TRIF, the clinical course was significantly attenuated compared to wild-type mice. This could be attributed to impaired NE-kappa B activation, as shown by the absence of nuclear translocation of ReIA with a decreased expression of IL-6, IL 12p40, and IL-17A. Remarkably, PO106-125-immunized TLR2(0/0) mice exhibited a delayed recovery as compared to TLR4(0/0) mice, which was because of an impaired T helper cell 2 polarization. Immunized TLR2(0/0) mice' were unable to induce OX40 and OX4OL by matrix metalloproteinase-2 on spLenic dendritic cells. Subsequently, M2 polarization was impaired and macrophages were unable to sufficiently induce T regulatory cells (T-regs). Thus, in the recovery phase, Tregs were significantly increased in TLR4(0/0) mice as compared to wild-type mice, whereas T-regs in immunized TLR2(0/0) mice were only slightly increased. Our data highlight the relevance of innate immunity and, especially, the tight interaction between the innate and the adaptive immune system, which should be considered for therapeutic approaches of autoimmune diseases

Emotionserleben, Emotionsregulation und strukturelle Störung bei Jugendlichen

Das Integrationsniveau psychischer Struktur steht mit psychischer Krankheit, sowie den Bereichen Emotionserleben und Emotionsregulation in engem Zusammenhang. Es lassen sich hierin basale Dimensionen vermuten, deren differenzierte Betrachtung wichtig ist, um zwischen einem passageren Geschehen und strukturellen Defiziten entscheiden zu können. Eine umfassende strukturelle Diagnostik wäre im Kinder- und Jugendpsychiatrischen Bereich wünschenswert, ist jedoch mit einem erheblichen Zeitaufwand verbunden, weshalb eine Umsetzung im klinischen Alltag oftmals nicht möglich ist. Ziel dieser Studie ist es, ein im klinischen Alltag ökonomisch durchführbares „Risiko-Screening“ zu erproben. So wurde anhand von 51 OPD-KJ Interviews mit Jugendlichen zwischen 14 und 17 Jahren eine Einschätzung des Strukturniveaus getätigt. Diese Jugendlichen füllten zusätzlich zwei Fragebögen aus: einen Fragebogen zum Emotionserleben und zur Emotionsregulation (EER) und den Fragebogen zur Persönlichkeitsorganisation (IPO-16). In der vorliegenden Arbeit wurde getestet, ob durch die Ergebnisse der beiden Selbstbeurteilungsverfahren vorhergesagt werden kann, bei welchen Jugendlichen weiterführende Strukturdiagnostik indiziert ist. Die Ergebnisse zeigen, dass das reduzierte Erleben positiver Emotionen und die Neigung, auf subjektiv als schwierig erlebte Gefühle mit einem Blackout zu reagieren, knapp 60 % der strukturell beeinträchtigten Jugendlichen korrekt zuordnete. Vierzig Prozent der strukturell vulnerablen Jugendlichen blieben jedoch durch dieses Screening unerkannt. Aus den Ergebnissen ist zu schließen, dass ein auf Selbstbeschreibungsverfahren beruhendes Screening im Rahmen einer ausführlichen Diagnostik zu Beginn der Behandlung im jugendpsychiatrischen Bereich nicht ohne zusätzliches klinisches Expertenurteil empfohlen werden kann.The level of structural integration, as well as the experience and regulation of emotions, are closely linked to mental illness. The differentiated consideration of these basic dimensions may therefore allow for a distinction between temporary clinical phenomena and more serious structural impairment. Especially in the department of childhood and adolescent psychiatry, it would thus be desirable to dispose of extensive diagnostics of structural impairment. However, this diagnostics takes up a considerable amount of time, which prevents the implementation in daily clinical practice. The aim of this study is to try out an economically practicable “risk-screening” for daily clinical practice. Therefore, 51 OPD-CA interviews were conducted with adolescents aged 14 to 17 years. In addition, the adolescents filled out two questionnaires: the questionnaire for the assessment of emotional experience and emotion regulation (EER) and the Inventory of Personality Organization (IPO). Logistic regression was used to estimate whether the results of self-report questionnaires predicted structural impairment. The results show that reduced experience of positive emotions, and the tendency to react with a black-out to the experience of subjectively difficult emotions, predicted just under 60% of structurally impaired adolescents. Forty percent of the adolescents with structural vulnerability however remained unnoticed by the screening. Overall, the results indicate that a screening, which is merely based on self-report questionnaires, is not sufficient for extensive diagnostics of structural impairment in childhood an adolescent psychiatry, and cannot be a substitute for the judgement of clinical experts.(VLID)4523631Version of recor

The HD(CP)2 Observational Prototype Experiment HOPE - an overview

The "HD(CP)2 Observational Prototype Experiment" (HOPE) was executed as a major 2-month field experiment in Jülich, Germany, performed in April and May 2013, followed by a smaller campaign in Melpitz, Germany in September 2013. HOPE has been designed to provide a critical evaluation of the new German community atmospheric Icosahedral non-hydrostatic (ICON) model at the scale of the model simulations and further to provide information on land-surface-atmospheric boundary layer exchange, cloud and precipitation processes as well as on sub-grid variability and microphysical properties that are subject to parameterizations. HOPE focuses on the onset of clouds and precipitation in the convective atmospheric boundary layer. The paper summarizes the instrument set-ups, the intensive observation periods as well as example results from both campaigns.HOPE-Jülich instrumentation included a radio sounding station, 4 Doppler lidars, 4 Raman lidars (3, 3, and 4 of these provide temperature, water vapor, and particle backscatter data, respectively), 1 water vapour differential absorption lidar, 3 cloud radars, 5 microwave radiometers, 3 rain radars, 6 sky imagers, 99 pyranometers, and 5 Sun photometers operated in synergy at different supersites. The HOPE-Melpitz campaign combined ground-based remote sensing of aerosols and clouds with helicopter- and balloon-based in-situ observations in the atmospheric column and at the surface.HOPE provided an unprecedented collection of atmospheric dynamical, thermodynamical, and micro- and macrophysical properties of aerosols, clouds and precipitation with high spatial and temporal resolution within a cube of approximately 10 × 10 × 10 km3. HOPE data will significantly contribute to our understanding of boundary layer dynamics and the formation of clouds and precipitation. The datasets are made available through a dedicated data portal