Untersuchung zum molekularen Mechanismus der chemisch Induzierten Resistenz in Gerste (Hordeum vulgare L.) gegenüber dem Echten Gerstenmehltaupilz (Blumeria graminis f. sp. hordei)

Durch die Applikation von chemischen Resistenzinduktoren kommt es in anfälligen Gerstensorten zu einer deutlich erhöhten systemischen Resistenz gegenüber dem Echten Gerstenmehltaupilz (Blumeria graminis f. sp. hordei, Bgh). Es sollten Gene identifiziert werden, die ursächlich an der Ausbildung dieser Form der Resistenz beteiligt sind und dadurch zur Aufklärung der beteiligten Signalwege beigetragen werden. Unter Verwendung von cDNA-Arrays wurden aus 1536 Genfragmenten 28 Gene identifiziert, die bislang nicht als chemisch induzierbar beschrieben waren. Für fünf dieser Gene wurde die Induzierbarkeit durch two step RT-PCRs bestätigt. Es handelt sich dabei um drei Gene mit bislang unbekannter Funktion, ein Gen mit Homologie zu Reticulon-ähnlichen Genen und eine Cysteinprotease. Ihre Beteiligung an der Resistenz der Gerste gegenüber Bgh wurde mittels transienten knock downs untersucht. Keines der unter¬suchten Gene scheint als Einzelfaktor für eine erfolgreiche Abwehrreaktion erforderlich zu sein. Ein sequenzhomologes Gerstengen zu AtEds5, welches in Arabidopsis ursächlich an der pathogen-induzierten SA-Produktion und SAR beteiligt ist, wurde ebenfalls in Gerste transient ausgeschaltet, um seine Beteiligung an der Resistenzausprägung zu überprüfen. Außerdem wurde die Funktion von HvSgt1 während der cIR in Gerste untersucht, da für Sgt1 in Gerste bereits eine Beteiligung an der R-Gen-vermittelten Resistenz gezeigt werden konnte. Des Weiteren wurden die chemisch induzierbaren Gene Bci2, ein blattspezifisches Thionin, Bci4, ein EF-hand Protein und Bci9, eine saure Phosphatase funktionell untersucht. Es standen stabil transgene Pflanzen zur Verfügung, die Bci4 überexprimieren, während Bci2 und Bci9 im transienten Transformationsassay untersucht wurden. Die transienten knock downs von Eds5 und Sgt1 haben keinen Einfluss auf die Penetrationseffizienz von Bgh. Für Bci4 und Bci9 konnte kein signifikanter Einfluss auf die Gersten-Mehltau Interaktion nachgewiesen werden, während der transiente knock down von Bci2 in BTH-behandelten Gerstenblättern zu einer erhöhten Suszeptibiltät gegenüber Bgh führt. Die Ergebnisse zeigen, dass sich cDNA-Arrays eher zur Expressionsanalyse, d. h. zur Analyse der Veränderungen in der Zelle, die zur metabolischen Adaption an die Induzierte Resistenz führen, aber nicht unbedingt zur Identifizierung ursächlich an der Resistenzausprägung beteiligter Gene in Gerste eignen. Dazu wäre die Analyse von Mutanten, wie in A. thaliana seit mehreren Jahren praktiziert, besser geeignet. Allerdings ist die Generierung von T-DNA-Insertionslinien in großem Maßstab in Gerste erst in den Anfängen.The application of chemical resistance inducers leads in susceptible barley lines to higher resistance against the infection with the barley powdery mildew fungus (Blumeria graminis f. sp. hordei, Bgh). Since the signal transduction pathways of the chemical induced resistance (cIR) in monocotyledonous plants are poorly understood, the aim of this work was to identify genes that are causally related to this kind of resistance. Using cDNA-arrays with 1536 gene fragments 28 new genes were identified, that were not described before to be chemical inducible. Five of them could be verified to be chemical inducible using the method of two-step RT-PCR. The functions of three of these genes have until now not been described, one shares homology with reticulon-like genes and one is a cysteine protease. The function of the five genes according the resistance of barley against Bgh was tested using a transient transformation system for silencing the gene expression. None of the tested genes as a single factor seems to have an influence on the pathogen response. A barley gene that shows highest homology to AtEds5, which causally involved in SA-production and SAR, was also transiently knocked down to check its involvement in resistance against Bgh. HvSgt1 was also tested using transient knock down, because it is involved in the R-gene mediated resistance in barley. Additionally the function of Bci2, a leaf-specific thionin, Bci4, an EF-hand protein and Bci9, an acid phosphatase were checked. Bci4 overexpressing cereal plants were available, while Bci2 and Bci9 were checked in transient transformation assays. Transient knock downs of the Eds5-homologue and HvSgt1 have no effect on penetration efficiency of Bgh. Overexpression of the three Bci-genes has also no effect on the resistance against Bgh. Just the transient knock down of Bci2 in BTH-treated barley leaves lead to higher susceptibility against Bgh. cDNA-arrays are a good tool for expression analysis in barley, e. g. for the changes concerning metabolic adaptation to the induced resistance, but they are not suitable for the identification of genes that are causally related to resistance in barley. A mutant screen as it is done in A. thaliana for many years would more appropriate. But the construction of mutant libraries in barley is still on the way

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Additional file 1: of BCL9L expression in pancreatic neoplasia with a focus on SPN: a possible explanation for the enigma of the benign neoplasia

Mean RNA expression Ratio derived from two independent experiments, normalized against GAPDH. (XLSX 12 kb

How do we turn surgical residents into safe intensive care unit clinicians? An Entrustable Professional Activities guided framework

10.1002/bjs.11949BRITISH JOURNAL OF SURGERY10711E491-E49

Table2_FACILITATE: A real-world, multicenter, prospective study investigating the utility of a rapid, fully automated real-time PCR assay versus local reference methods for detecting epidermal growth factor receptor variants in NSCLC.DOCX

Accurate testing for epidermal growth factor receptor (EGFR) variants is essential for informing treatment decisions in non-small cell lung cancer (NSCLC). Automated diagnostic workflows may allow more streamlined initiation of targeted treatments, where appropriate, while comprehensive variant analysis is ongoing. FACILITATE, a real-world, prospective, multicenter, European study, evaluated performance and analytical turnaround time of the Idylla™ EGFR Mutation Test compared with local reference methods. Sixteen sites obtained formalin-fixed paraffin-embedded biopsy samples with ≥ 10% neoplastic cells from patients with NSCLC. Consecutive 5 μm sections from patient samples were tested for clinically relevant NSCLC-associated EGFR variants using the Idylla™ EGFR Mutation Test and local reference methods; performance (concordance) and analytical turnaround time were compared. Between January 2019 and November 2020, 1,474 parallel analyses were conducted. Overall percentage agreement was 97.7% [n = 1,418; 95% confidence interval (CI): 96.8–98.3], positive agreement, 87.4% (n = 182; 95% CI: 81.8–91.4) and negative agreement, 99.2% (n = 1,236; 95% CI: 98.5–99.6). There were 38 (2.6%) discordant cases. Ninety percent of results were returned with an analytical turnaround time of within 1 week using the Idylla™ EGFR Mutation Test versus ∼22 days using reference methods. The Idylla™ EGFR Mutation Test performed well versus local methods and had shorter analytical turnaround time. The Idylla™ EGFR Mutation Test can thus support application of personalized medicine in NSCLC.</p

Table1_FACILITATE: A real-world, multicenter, prospective study investigating the utility of a rapid, fully automated real-time PCR assay versus local reference methods for detecting epidermal growth factor receptor variants in NSCLC.DOCX

Accurate testing for epidermal growth factor receptor (EGFR) variants is essential for informing treatment decisions in non-small cell lung cancer (NSCLC). Automated diagnostic workflows may allow more streamlined initiation of targeted treatments, where appropriate, while comprehensive variant analysis is ongoing. FACILITATE, a real-world, prospective, multicenter, European study, evaluated performance and analytical turnaround time of the Idylla™ EGFR Mutation Test compared with local reference methods. Sixteen sites obtained formalin-fixed paraffin-embedded biopsy samples with ≥ 10% neoplastic cells from patients with NSCLC. Consecutive 5 μm sections from patient samples were tested for clinically relevant NSCLC-associated EGFR variants using the Idylla™ EGFR Mutation Test and local reference methods; performance (concordance) and analytical turnaround time were compared. Between January 2019 and November 2020, 1,474 parallel analyses were conducted. Overall percentage agreement was 97.7% [n = 1,418; 95% confidence interval (CI): 96.8–98.3], positive agreement, 87.4% (n = 182; 95% CI: 81.8–91.4) and negative agreement, 99.2% (n = 1,236; 95% CI: 98.5–99.6). There were 38 (2.6%) discordant cases. Ninety percent of results were returned with an analytical turnaround time of within 1 week using the Idylla™ EGFR Mutation Test versus ∼22 days using reference methods. The Idylla™ EGFR Mutation Test performed well versus local methods and had shorter analytical turnaround time. The Idylla™ EGFR Mutation Test can thus support application of personalized medicine in NSCLC.</p

Table3_FACILITATE: A real-world, multicenter, prospective study investigating the utility of a rapid, fully automated real-time PCR assay versus local reference methods for detecting epidermal growth factor receptor variants in NSCLC.DOCX

Accurate testing for epidermal growth factor receptor (EGFR) variants is essential for informing treatment decisions in non-small cell lung cancer (NSCLC). Automated diagnostic workflows may allow more streamlined initiation of targeted treatments, where appropriate, while comprehensive variant analysis is ongoing. FACILITATE, a real-world, prospective, multicenter, European study, evaluated performance and analytical turnaround time of the Idylla™ EGFR Mutation Test compared with local reference methods. Sixteen sites obtained formalin-fixed paraffin-embedded biopsy samples with ≥ 10% neoplastic cells from patients with NSCLC. Consecutive 5 μm sections from patient samples were tested for clinically relevant NSCLC-associated EGFR variants using the Idylla™ EGFR Mutation Test and local reference methods; performance (concordance) and analytical turnaround time were compared. Between January 2019 and November 2020, 1,474 parallel analyses were conducted. Overall percentage agreement was 97.7% [n = 1,418; 95% confidence interval (CI): 96.8–98.3], positive agreement, 87.4% (n = 182; 95% CI: 81.8–91.4) and negative agreement, 99.2% (n = 1,236; 95% CI: 98.5–99.6). There were 38 (2.6%) discordant cases. Ninety percent of results were returned with an analytical turnaround time of within 1 week using the Idylla™ EGFR Mutation Test versus ∼22 days using reference methods. The Idylla™ EGFR Mutation Test performed well versus local methods and had shorter analytical turnaround time. The Idylla™ EGFR Mutation Test can thus support application of personalized medicine in NSCLC.</p