LANDSCAPE AESTHETICS AS AN INDICATOR FOR SOCIAL SUSTAINABILITY OF CROP ROTATIONS

The goal of this study was to evaluate different crop rotations in terms of landscape aesthetics. Klostergut Scheyern has been used as an example, since its current crop rotation must be changed. Therefore, people living in the neighbourhood of Klostergut Scheyern have been shown photos of each crop at different periods of the year. Additionally, a questionnaire containing quantitative, as well as qualitative questions has been developed. Each crop has got a grade; the best one being 1 up to the worst one being 4. People have been allowed to justify their ranking. Also, the use of agricultural machines has been presented on photos. For each crop, participants could decide whether they liked or disliked the working steps. In summary, all crops have been evaluated rather positively. The grades reach from 1.4 to 2.6. The use of machines has hardly ever influenced people’s opinions. Taking qualitative comments into consideration, the survey revealed the difficulty to separate purely visual aspects from moral attitudes towards a certain crop

Evaluierung und Optimierung biologischer Verfahren zur Regulierung des Pflaumenwicklers (Cydia funebrana) und der Monilia-Krankheit im ökologischen Steinobstanbau

Im Verbundforschungsprojekt FKZ 06OE198, FKZ 06OE057 und FKZ 06OE348 wurden Möglichkeiten der Regulierung des Pflaumenwicklers (Cydia funebrana) an Zwetschgen in Labor-, Halbfreiland und Freilandversuchen in Geisenheim, Darmstadt, Klein-Altendorf und Weinsberg untersucht. Labor- und Freilandversuche zur Regulierung der Monilia-Krankheit wurden in Geisenheim, Weinsberg und Dresden durchgeführt. Sauerkirschsorten-Versuche sind in Weinsberg und Dresden aufgepflanzt. Umfangreiche Freilandversuche zur Verwirrung des Pflaumenwicklers mit „Isomate OFM Rosso“ in Baden-Württemberg und Rheinland-Pfalz bestätigten mit Wirkungsgraden von bis zu 70 % die Wirksamkeit dieser Methode bei ausreichend großen Flächen und niedrigem Befallsniveau. Für den kleinparzellierten Anbau und bei höherem Befallsdruck wurden entomopathogene Nematoden im Freiland gegen überwinternde Larven mit heterogenen Wirkungsgraden erprobt. Es gelang nicht Pflaumenwickler ganzjährig unter Laborbedingen zu züchten, jedoch wurden mit Tieren aus Freilandsammlungen bzw. deren Eiern nach der Überwinterung Versuche zur Infektion der neonaten Larven mit dem Apfelwicklergranulosevirus, Eiparasitierungsversuche mit Trichogramma cacoeciae, zu Applikationen mit NeemAzal-T/S und zum Einsatz von insektenpathogen Pilzen gegen überwinternde Larven durchgeführt. Bei weiterführenden Versuchen im Freiland wurden mit dem Apfelwicklergranulosevirus V15 sehr heterogene Ergebnisse erzielt. Der Einsatz von Trichogramma in Form von Trichokarten ergab Wirkungsgrade von bis zu 47 %. Eine bessere Verteilung der Trichogramma in der Anlage sollte mit einer spritzfähigen Formulierung erreicht werden, dafür liegen jedoch erst einjährige Ergebnisse vor. Für die Bekämpfung der überwinternden Pflaumenwickler mit insektenpathogenen Pilzen stellte sich Isaria fumosorosea, als am geeignetsten heraus. Jedoch gestaltete sich bisher ein Nachweis der Wirksamkeit des mit dem Pilz infizierten Mulches im Freiland sehr schwierig. Die Pflaumenwicklerlarve sucht sich zur Überwinterung kleinste Ritzen am Stamm aus. Mit Wellpappe, die um den Stamm gewickelt wurde, konnte den Larven ein attraktives Überwinterungsquartier angeboten werden. Durch Entfernen und Verbrennen der Pappringe im Winter kann der Befallsdruck im Folgejahr minimiert werden. Für die Monilia-Regulierung an Zwetschgen konnte im Labor unter einer Vielzahl an getesteten Präparaten lediglich Serenade mit einer antagonistischen Wirkung überzeugen. Es hat jedoch keine Zulassung als Pflanzenschutzmittel im Steinobst. Versuche im Freiland ergaben außerdem, dass das konsequente Eliminieren von möglichen Infektionsquellen durch das Entfernen von Monilia-Nestern vor der Ernte und von Fruchtmumien nach der Ernte letztlich bessere Effekte erzielte, als Behandlungen mit mittelgut wirksamen Pflanzenstärkungsmitteln. Für die Behandlung der Monilia-Spitzendürre an Sauerkirschen wurden verschiedene Präparate im Freiland überprüft. Die Handelsprodukte BoniProtect® forte und BoniProtect® reduzierten tendenziell die Blüteninfektionen. Da für SPU 2720 und Cuprozin bisher nur aus einem Jahr Versuchsergebnisse vorliegen, ist keine abschließende Beurteilung möglich. Jedoch lässt sich bereits durch den konsequenten Rückschnitt befallener Triebe der Befallsdruck erheblich reduzieren. An beiden Standorten erscheinen die Sauerkirschsorten ‘Morina‘, ‘Jade‘, ‘Coralin‘ und ‘Safir‘ als geeignet für den ökologischen Anbau, in Weinsberg zusätzlich ‘Topas‘ und ‘Ungarische Traubige‘. Allerdings war der Monilia-Druck nur in wenigen Jahren hoch. Bis jetzt wurden keine Infektionen mit Gloeosporium sp., einer weiteren bedrohlichen Krankheit im ökologischen Sauerkirschanbau, beobachtet

Future Directions in the Diagnosis and Treatment of APDS and IEI: a Survey of German IEI Centers

IntroductionThe diagnosis and treatment of inborn errors of immunity (IEI) is a major challenge as the individual conditions are rare and often characterized by a variety of symptoms, which are often non disease-specific. Ideally, patients are treated in dedicated centers by physicians who specialize in the management of primary immune disorders. In this study, we used the example of Activated PI3Kδ syndrome (APDS), a rare IEI with an estimated prevalence of 1:1,000,000. We conducted surveys by questionnaire and interviewed physicians at different IEI centers in Germany.MethodsWe queried structural aspects of IEI care in Germany, diagnostic procedures in IEI care (including molecular diagnostics), distribution of APDS patients, APDS symptoms and severity, treatment algorithms in APDS, the role of stem cell transplantation and targeted therapies in IEI with focus on APDS. We were especially interested in how genetic diagnostics may influence treatment decisions, e.g. with regard to targeted therapies.Results/discussionMost centers care for both pediatric and adult patients. A total of 28 APDS patients are currently being treated at the centers we surveyed. Patient journeys vary considerably, as does severity of disease. Genetic diagnosis continues to gain importance - whole genome sequencing is likely to become routine in IEI in the next few years. According to the experts interviewed, stem cell transplantation and - with new molecules being approved - targeted therapies, will gain in importance for the treatment of APDS and IEI in general

Oral insulin immunotherapy in children at risk for type 1 diabetes in a randomised controlled trial

AIMS/HYPOTHESIS Oral administration of antigen can induce immunological tolerance. Insulin is a key autoantigen in childhood type 1 diabetes. Here, oral insulin was given as antigen-specific immunotherapy before the onset of autoimmunity in children from age 6~months to assess its safety and immune response actions on immunity and the gut microbiome. METHODS A phase I/II randomised controlled trial was performed in a single clinical study centre in Germany. Participants were 44 islet autoantibody-negative children aged 6~months to 2.99~years who had a first-degree relative with type 1 diabetes and a susceptible HLA DR4-DQ8-containing genotype. Children were randomised 1:1 to daily oral insulin (7.5~mg with dose escalation to 67.5~mg) or placebo for 12~months using a web-based computer system. The primary outcome was immune efficacy pre-specified as induction of antibody or T cell responses to insulin and measured in a central treatment-blinded laboratory. RESULTS Randomisation was performed in 44 children. One child in the placebo group was withdrawn after the first study visit and data from 22 insulin-treated and 21 placebo-treated children were analysed. Oral insulin was well tolerated with no changes in metabolic variables. Immune responses to insulin were observed in children who received both insulin (54.5%) and placebo (66.7%), and the trial did not demonstrate an effect on its primary outcome (p = 0.54). In exploratory analyses, there was preliminary evidence that the immune response and gut microbiome were modified by the INS genotype Among children with the type 1 diabetes-susceptible INS genotype (n = 22), antibody responses to insulin were more frequent in insulin-treated (72.7%) as compared with placebo-treated children (18.2%; p = 0.03). T cell responses to insulin were modified by treatment-independent inflammatory episodes. CONCLUSIONS/INTERPRETATION The study demonstrated that oral insulin immunotherapy in young genetically at-risk children was safe, but was not associated with an immune response as predefined in the trial primary outcome. Exploratory analyses suggested that antibody responses to oral insulin may occur in children with a susceptible INS genotype, and that inflammatory episodes may promote the activation of insulin-responsive T cells. TRIAL REGISTRATION Clinicaltrials.gov NCT02547519 FUNDING: The main funding source was the German Center for Diabetes Research (DZD e.V.)

Targeting MYC effector functions in pancreatic cancer by inhibiting the ATPase RUVBL1/2

Objective The hallmark oncogene MYC drives the progression of most tumours, but direct inhibition of MYC by a small-molecule drug has not reached clinical testing. MYC is a transcription factor that depends on several binding partners to function. We therefore explored the possibility of targeting MYC via its interactome in pancreatic ductal adenocarcinoma (PDAC). Design To identify the most suitable targets among all MYC binding partners, we constructed a targeted shRNA library and performed screens in cultured PDAC cells and tumours in mice. Results Unexpectedly, many MYC binding partners were found to be important for cultured PDAC cells but dispensable in vivo. However, some were also essential for tumours in their natural environment and, among these, the ATPases RUVBL1 and RUVBL2 ranked first. Degradation of RUVBL1 by the auxin-degron system led to the arrest of cultured PDAC cells but not untransformed cells and to complete tumour regression in mice, which was preceded by immune cell infiltration. Mechanistically, RUVBL1 was required for MYC to establish oncogenic and immunoevasive gene expression identifying the RUVBL1/2 complex as a druggable vulnerability in MYC-driven cancer. Conclusion One implication of our study is that PDAC cell dependencies are strongly influenced by the environment, so genetic screens should be performed in vitro and in vivo. Moreover, the auxin-degron system can be applied in a PDAC model, allowing target validation in living mice. Finally, by revealing the nuclear functions of the RUVBL1/2 complex, our study presents a pharmaceutical strategy to render pancreatic cancers potentially susceptible to immunotherapy

Multicenter Collaborative Study to Optimize Mass Spectrometry Workflows of Clinical Specimens

The foundation for integrating mass spectrometry (MS)-based proteomics into systems medicine is the development of standardized start-to-finish and fit-for-purpose workflows for clinical specimens. An essential step in this pursuit is to highlight the common ground in a diverse landscape of different sample preparation techniques and liquid chromatography-mass spectrometry (LC-MS) setups. With the aim to benchmark and improve the current best practices among the proteomics MS laboratories of the CLINSPECT-M consortium, we performed two consecutive round-robin studies with full freedom to operate in terms of sample preparation and MS measurements. The six study partners were provided with two clinically relevant sample matrices: plasma and cerebrospinal fluid (CSF). In the first round, each laboratory applied their current best practice protocol for the respective matrix. Based on the achieved results and following a transparent exchange of all lab-specific protocols within the consortium, each laboratory could advance their methods before measuring the same samples in the second acquisition round. Both time points are compared with respect to identifications (IDs), data completeness, and precision, as well as reproducibility. As a result, the individual performances of participating study centers were improved in the second measurement, emphasizing the effect and importance of the expert-driven exchange of best practices for direct practical improvements

The Adaptor Molecule Nck Localizes the WAVE Complex to Promote Actin Polymerization during CEACAM3-Mediated Phagocytosis of Bacteria

Background: CEACAM3 is a granulocyte receptor mediating the opsonin-independent recognition and phagocytosis of human-restricted CEACAM-binding bacteria. CEACAM3 function depends on an intracellular immunoreceptor tyrosine-based activation motif (ITAM)-like sequence that is tyrosine phosphorylated by Src family kinases upon receptor engagement. The phosphorylated ITAM-like sequence triggers GTP-loading of Rac by directly associating with the guanine nucleotide exchange factor (GEF) Vav. Rac stimulation in turn is critical for actin cytoskeleton rearrangements that generate lamellipodial protrusions and lead to bacterial uptake. Principal Findings: In our present study we provide biochemical and microscopic evidence that the adaptor proteins Nck1 and Nck2, but not CrkL, Grb2 or SLP-76, bind to tyrosine phosphorylated CEACAM3. The association is phosphorylation-dependent and requires the Nck SH2 domain. Overexpression of the isolated Nck1 SH2 domain, RNAi-mediated knock-down of Nck1, or genetic deletion of Nck1 and Nck2 interfere with CEACAM3-mediated bacterial internalization and with the formation of lamellipodial protrusions. Nck is constitutively associated with WAVE2 and directs the actin nucleation promoting WAVE complex to tyrosine phosphorylated CEACAM3. In turn, dominant-negative WAVE2 as well as shRNA-mediated knock-down of WAVE2 or the WAVE-complex component Nap1 reduce internalization of bacteria. Conclusions: Our results provide novel mechanistic insight into CEACAM3-initiated phagocytosis. We suggest that the CEACAM3 ITAM-like sequence is optimized to co-ordinate a minimal set of cellular factors needed to efficiently trigger actin-based lamellipodial protrusions and rapid pathogen engulfment

Systematic multi-omics cell line profiling uncovers principles of Ewing sarcoma fusion oncogene-mediated gene regulation

Ewing sarcoma (EwS) is characterized by EWSR1-ETS fusion transcription factors converting polymorphic GGAA microsatellites (mSats) into potent neo-enhancers. Although the paucity of additional mutations makes EwS a genuine model to study principles of cooperation between dominant fusion oncogenes and neo-enhancers, this is impeded by the limited number of well-characterized models. Here we present the Ewing Sarcoma Cell Line Atlas (ESCLA), comprising whole-genome, DNA methylation, transcriptome, proteome, and chromatin immunoprecipitation sequencing (ChIP-seq) data of 18 cell lines with inducible EWSR1-ETS knockdown. The ESCLA shows hundreds of EWSR1-ETS-targets, the nature of EWSR1-ETS-preferred GGAA mSats, and putative indirect modes of EWSR1-ETS-mediated gene regulation, converging in the duality of a specific but plastic EwS signature. We identify heterogeneously regulated EWSR1-ETS-targets as potential prognostic EwS biomarkers. Our freely available ESCLA (http://r2platform.com/escla/) is a rich resource for EwS research and highlights the power of comprehensive datasets to unravel principles of heterogeneous gene regulation by chimeric transcription factors

Curation and expansion of Human Phenotype Ontology for defined groups of inborn errors of immunity.

BACKGROUND: Accurate, detailed, and standardized phenotypic descriptions are essential to support diagnostic interpretation of genetic variants and to discover new diseases. The Human Phenotype Ontology (HPO), extensively used in rare disease research, provides a rich collection of vocabulary with standardized phenotypic descriptions in a hierarchical structure. However, to date, the use of HPO has not yet been widely implemented in the field of inborn errors of immunity (IEIs), mainly due to a lack of comprehensive IEI-related terms. OBJECTIVES: We sought to systematically review available terms in HPO for the depiction of IEIs, to expand HPO, yielding more comprehensive sets of terms, and to reannotate IEIs with HPO terms to provide accurate, standardized phenotypic descriptions. METHODS: We initiated a collaboration involving expert clinicians, geneticists, researchers working on IEIs, and bioinformaticians. Multiple branches of the HPO tree were restructured and extended on the basis of expert review. Our ontology-guided machine learning coupled with a 2-tier expert review was applied to reannotate defined subgroups of IEIs. RESULTS: We revised and expanded 4 main branches of the HPO tree. Here, we reannotated 73 diseases from 4 International Union of Immunological Societies-defined IEI disease subgroups with HPO terms. We achieved a 4.7-fold increase in the number of phenotypic terms per disease. Given the new HPO annotations, we demonstrated improved ability to computationally match selected IEI cases to their known diagnosis, and improved phenotype-driven disease classification. CONCLUSIONS: Our targeted expansion and reannotation presents enhanced precision of disease annotation, will enable superior HPO-based IEI characterization, and hence benefit both IEI diagnostic and research activities