Switchgrass (\u3ci\u3ePanicum virgatum\u3c/i\u3e L.) polyubiquitin gene (\u3ci\u3ePvUbi1\u3c/i\u3e and \u3ci\u3ePvUbi2\u3c/i\u3e) promoters for use in plant transformation

Abstract Background The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (Panicum virgatum L.) ubiquitin genes (PvUbi1 and PvUbi2) were cloned and characterized. Reporter constructs were produced containing the isolated 5\u27 upstream regulatory regions of the coding sequences (i.e. PvUbi1 and PvUbi2 promoters) fused to the uidA coding region (GUS) and tested for transient and stable expression in a variety of plant species and tissues. Results PvUbi1 consists of 607 bp containing cis-acting regulatory elements, a 5\u27 untranslated region (UTR) containing a 93 bp non-coding exon and a 1291 bp intron, and a 918 bp open reading frame (ORF) that encodes four tandem, head -to-tail ubiquitin monomer repeats followed by a 191 bp 3\u27 UTR. PvUbi2 consists of 692 bp containing cis-acting regulatory elements, a 5\u27 UTR containing a 97 bp non-coding exon and a 1072 bp intron, a 1146 bp ORF that encodes five tandem ubiquitin monomer repeats and a 183 bp 3\u27 UTR. PvUbi1 and PvUbi2 were expressed in all examined switchgrass tissues as measured by qRT-PCR. Using biolistic bombardment, PvUbi1 and PvUbi2 promoters showed strong expression in switchgrass and rice callus, equaling or surpassing the expression levels of the CaMV 35S, 2x35S, ZmUbi1, and OsAct1 promoters. GUS staining following stable transformation in rice demonstrated that the PvUbi1 and PvUbi2 promoters drove expression in all examined tissues. When stably transformed into tobacco (Nicotiana tabacum), the PvUbi2+3 and PvUbi2+9 promoter fusion variants showed expression in vascular and reproductive tissues. Conclusions The PvUbi1 and PvUbi2 promoters drive expression in switchgrass, rice and tobacco and are strong constitutive promoter candidates that will be useful in genetic transformation of monocots and dicots

Read-through Activation of Transcription in a Cellular Genomic Context

Read-through transcription from the adjacent E1a gene region is required for wild-type (wt) activity of the downstream adenovirus E1b promoter early after infection (read-through activation). However, whether a cellular chromosomal template can support read-through activation is not known. To address this issue, read-through activation was evaluated in the context of stably expressed templates in transfected cells. Inhibition of read-through transcription by insertion of a transcription termination sequence between the E1a and E1b promoters reduced downstream gene expression from stably integrated templates. The results indicate that the mechanism of read-through activation does not depend on the structure of early adenovirus nucleoprotein complexes, a structure that is likely to be different from that of cellular chromatin. Accordingly, this regulatory interaction could participate in the coordinated control of the expression of closely linked cellular genes

Switchgrass (Panicum virgatum L.) polyubiquitin gene (PvUbi1 and PvUbi2) promoters for use in plant transformation

<p>Abstract</p> <p>Background</p> <p>The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (<it>Panicum virgatum </it>L.) ubiquitin genes (<it>PvUbi1 </it>and <it>PvUbi2</it>) were cloned and characterized. Reporter constructs were produced containing the isolated 5' upstream regulatory regions of the coding sequences (i.e. <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters) fused to the <it>uidA </it>coding region (<it>GUS</it>) and tested for transient and stable expression in a variety of plant species and tissues.</p> <p>Results</p> <p><it>PvUbi1 </it>consists of 607 bp containing <it>cis</it>-acting regulatory elements, a 5' untranslated region (UTR) containing a 93 bp non-coding exon and a 1291 bp intron, and a 918 bp open reading frame (ORF) that encodes four tandem, head -to-tail ubiquitin monomer repeats followed by a 191 bp 3' UTR. <it>PvUbi2 </it>consists of 692 bp containing <it>cis</it>-acting regulatory elements, a 5' UTR containing a 97 bp non-coding exon and a 1072 bp intron, a 1146 bp ORF that encodes five tandem ubiquitin monomer repeats and a 183 bp 3' UTR. <it>PvUbi1 </it>and <it>PvUbi2 </it>were expressed in all examined switchgrass tissues as measured by qRT-PCR. Using biolistic bombardment, <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters showed strong expression in switchgrass and rice callus, equaling or surpassing the expression levels of the CaMV <it>35S, 2x35S, ZmUbi1</it>, and <it>OsAct1 </it>promoters. GUS staining following stable transformation in rice demonstrated that the <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters drove expression in all examined tissues. When stably transformed into tobacco (<it>Nicotiana tabacum</it>), the <it>PvUbi2+3 </it>and <it>PvUbi2+9 </it>promoter fusion variants showed expression in vascular and reproductive tissues.</p> <p>Conclusions</p> <p>The <it>PvUbi1 </it>and <it>PvUbi2 </it>promoters drive expression in switchgrass, rice and tobacco and are strong constitutive promoter candidates that will be useful in genetic transformation of monocots and dicots.</p

Monte Carlo simulation of aggregate morphology for simultaneous coagulation and sintering

A model for simulation of the three-dimensional morphology of nano-structured aggregates formed by concurrent coagulation and sintering is presented. Diffusion controlled cluster-cluster aggregation is assumed to be the prevailing coagulation mechanism which is implemented using a Monte-Carlo algorithm. Sintering is modeled as a successive overlapping of spherical primary particles, which are allowed to grow as to preserve overall mass. Simulations are characterized by individual ratios tau of characteristic collision to fusion time. A number of resulting aggregate-structures is displayed and reveals structure formation by coagulation and sintering for different values of tau. These aggregates are described qualitatively and quantitatively by their mass fractal dimension D-f and radius of gyration. The fractal dimension increases from 1.86 for pure aggregation to approximate to 2.75 for equal characteristic time scales. As sintering turns out to be more and more relevant, increasingly compact aggregates start to form and the radius of gyration decreases significantly. The simulation results clearly reveal a strong dependence of the fractal dimension on the kinetics of the concurrent coagulation and sintering processes. Considering appropriate values of D-f in aerosol process simulations may therefore be important in many cases

Surveillance postoperativer Wundinfektionen: methodischer Vergleich von IQTIG- und KISS-Strategie

Aim: In 2017, the Institute for Quality Assurance and Transparency in Healthcare (IQTIG) introduced a quality assurance system for the surveillance of surgical site infections (SSI) on behalf the Federal Joint Committee. The establishment of the new system was made in parallel to existing methods, such as the "Krankenhaus-Infektions-Surveillance-System" (KISS). The aim of this work was to perform a comparative analysis.Methods: All 2,233 cases at the University Medical Center Goettingen requiring an assessment of the presence of SSI as part of the IQTIG procedure in 2018 and 2019 were evaluated retrospectively according to the KISS protocol.Results: In total, 2,050 patients were included in the comparative evaluation. Overall, 1,779 (79.7%) had a surgical anamnesis (surgery during the stay or in the past), and 1,716 (83.7%) showed identical results for both surveillance strategies. Different results were found for 334 patients (16.3%), with 160 of these (7.8%) positive for SSI according to IQTIG and 174 (8.5%) positive for KISS. Risk factors were identified for a discordant assessment between the methods. Conclusion: The congruence of the two strategies was consistently high over the study period. There is evidence that the efficiency of the documentation algorithm can be increased without the loss of documentation of SSI, while preserving the precision of the documentation through training.Zielsetzung: Zur Surveillance postoperativer Wundinfektionen wurde 2017 im Auftrag des Gemeinsamen Bundesausschusses ein sektorenübergreifendes Qualitätssicherungsverfahren durch das Institut für Qualitätssicherung und Transparenz im Gesundheitswesen (IQTIG) eingeführt (Vermeidung nosokomialer Infektionen - postoperati ve Wu ndinfektionen, QS WI ). Die Etablierung erfolgte parallel zu bestehenden Methoden wie z.B. des Krankenhaus-Infektions-Surveillance-Systems (KISS). Das Ziel dieser Arbeit war die vergleichende Analyse dieser beiden Methoden.Methodik: Alle 2.233 Patientenfälle, die im Rahmen des IQTIG QS-Verfahrens 2018 und 2019 an der Universitätsmedizin Göttingen (UMG) eine Bewertung bzgl. Vorhandenseins postoperativer Wundinfektionen erforderten, wurden retrospektiv gemäß den Vorgaben des KISS-Protokolls nachbeurteilt und abschließend mit den Ergebnissen der IQTIG-Methodik verglichen.Ergebnisse: 2.050 Fälle konnten in die vergleichende Auswertung einbezogen werden. 1.779 (79,7%) wiesen einen primären chirurgischen Bezug auf (OP während des Aufenthalts/zuvor/ggf. extern). 1.716 (83,7%) zeigten identische Resultate bei den Bewertungen durch die beiden Surveillance-Systeme. Bei 334 (16,3%) kam es zu abweichenden Ergebnissen, wobei sich 160 (7,8%) mit einer positiven Wundinfektionsbeurteilung nach IQTIG und 174 (8,5%) mit einer positiven Bewertung nach KISS ergaben. Es fanden sich verschiedene Risikofaktoren für eine unterschiedliche Beurteilung zwischen den Verfahren.Fazit: Die Kongruenz beider Strategien zeigt sich über den Beobachtungszeitraum konstant. Es konnten Hinweise zur Optimierung der Effizienz des potenziell zu breit angelegten Auslösealgorithmus im QS WI-Verfahren ohne Verlust der Dokumentation postoperativer Wundinfektionen identifiziert werden Zudem zeigt sich Potenzial zur Steigerung der Dokumentationspräzision durch Schulungsmaßnahmen

Use of a serious game to teach infectious disease management in medical school: effectiveness and transfer to a clinical examination

PURPOSE: Physicians of all specialties must be familiar with important principles of infectious diseases, but curricular time for this content is limited and clinical teaching requires considerable resources in terms of available patients and teachers. Serious games are scalable interventions that can help standardize teaching. This study assessed whether knowledge and skills acquired in a serious game translate to better performance in a clinical examination. METHODS: Fifth-year undergraduate medical students (n = 100) at Goettingen Medical School were randomized to three groups receiving different levels of exposure to virtual patients presenting with signs and symptoms of either infective endocarditis or community-acquired pneumonia in a serious game simulating an accident and emergency department. Student performance was assessed based on game logfiles and an objective standardized clinical examination (OSCE). RESULTS: Higher exposure to virtual patients in the serious game did not result in superior OSCE scores. However, there was good agreement between student performance in the OSCE and in game logfiles (r = 0.477, p = 0.005). An Item Response Theory analysis suggested that items from the serious game covered a wider range of ability, thus better differentiating between students within a given cohort. CONCLUSION: Repeated exposure to virtual patients with infectious diseases in a serious game did not directly impact on exam performance but game logfiles might be good and resource-sparing indicators of student ability. One advantage of using serious games in medical education is standardized content, a lower inhibition threshold to learn, and a need of less staff time compared to small-group clinical teaching