Sensitive and powerful single-cell RNA sequencing using mcSCRB-seq

Single-cell RNA sequencing (scRNA-seq) has emerged as a central genome-wide method to characterize cellular identities and processes. Consequently, improving its sensitivity, flexibility, and cost-efficiency can advance many research questions. Among the flexible platebased methods, single-cell RNA barcoding and sequencing (SCRB-seq) is highly sensitive and efficient. Here, we systematically evaluate experimental conditions of this protocol and find that adding polyethylene glycol considerably increases sensitivity by enhancing cDNA synthesis. Furthermore, using Terra polymerase increases efficiency due to a more even cDNA amplification that requires less sequencing of libraries. We combined these and other improvements to develop a scRNA-seq library protocol we call molecular crowding SCRB-seq (mcSCRB-seq), which we show to be one of the most sensitive, efficient, and flexible scRNA-seq methods to date

Prime-seq, efficient and powerful bulk RNA sequencing.

Cost-efficient library generation by early barcoding has been central in propelling single-cell RNA sequencing. Here, we optimize and validate prime-seq, an early barcoding bulk RNA-seq method. We show that it performs equivalently to TruSeq, a standard bulk RNA-seq method, but is fourfold more cost-efficient due to almost 50-fold cheaper library costs. We also validate a direct RNA isolation step, show that intronic reads are derived from RNA, and compare cost-efficiencies of available protocols. We conclude that prime-seq is currently one of the best options to set up an early barcoding bulk RNA-seq protocol from which many labs would profit

Cost effectiveness of bilateral risk-reducing mastectomy and salpingo-oophorectomy

Background Growing demand for risk-reducing surgery in individuals with inherited susceptibility to cancer leads to the question whether these procedures are cost effective for the executing hospitals. This study compared the clinical costs for bilateral risk-reducing mastectomy (BRRM) with and without different types of reconstruction, risk-reducing salpingo-oophorectomy (RRSO), and their combinations with corresponding reimbursements in the statutory health-care system in Germany. Patients and methods Real total costs of care for BRRM with and without reconstruction, RRSO, and their combinations were calculated as the sum of all personnel and technical costs. These costs calculated in a German University hospital were compared with the sum of all reimbursements in the German DRG-based health-care system. Results While sole RRSO, BRRM without reconstruction, and BRRM with secondary DIEP (deep inferior epigastric perforator)—reconstruction still result in a small benefit, we even found shortfalls for the hospital with all other prophylactic operations under consideration. The calculated deficits were especially high for BRRM with implant-based breast reconstruction and for combined operations when the risk reduction is achieved with a minimum of separate operations. Conclusions Risk-reducing surgery in BRCA-mutation carriers is frequently not cost-covering for the executing hospitals in the German health-care system. Thus, appropriate concepts are required to ensure a nationwide care

Referenzmessverfahren fuer die Bestimmung von Schadstoffemissionen an Abfallverbrennungsanlagen. Teilvorhaben 4 Untersuchungen zur Gesamtkohlenstoff- und Kohlenmonoxid-Bestimmung in Abgasen von Abfallverbrennungsanlagen

a) In order to get informations about the spectrum of organic components in the stack gas of waste incineration plants (AVA) a literature review was carried out (only for components emitted in higher concentrations than 0,01 mg/m3). In the few existing publications there were no indications to components or component classes which are characteristic for AVAs. b) By reference measurements by conventional methods for total gaseous organic carbon (adsorption-[ADS] and FID-method) in connection with determination of single organic compounds the usability of total gaseous organic carbon measurement at AVAs should be tested. The ADS-method proved to be not suitable because of unplausible high measurement values (e.g. disturbance by interferences. The FID-method can be used in the area of a convention; as there is no homogenuous component spectrum in the stack gas, the exact conversion of the measurement values on the organic carbon is not possible. c) As there is no measurement method of total gaseous organic carbon comparing analysis known, the reference methods for the calibration of FID should be investigated. A method to determine response factors at installed FIDs in the AVA exhaust is described. d) For the determination of the component CO at AVAs the Iodine pentoxide method was investigated. The statistic evaluation of the measurement results shows that the method is not suitable for exact test gas analysis. Due to the high straying at investigations at real exhaust gas and the unsuitable usability the method can't be recommended for the usage at plants. (orig.)a) Mit einer Literaturrecherche sollte versucht werden, Informationen ueber das organische Stoffspektrum an Abfallverbrennungsanlagen (AVA) zu erhalten (nur Stoffe, die in hoeheren Konzentrationen als 0,01 mg/m3 emittiert werden). Es konnten in den wenigen Veroeffentlichungen keine Hinweise auf Stoffe oder Stoffklassen gefunden werden, die fuer AVA's charakteristisch sind. b) Ueber Vergleichsmessungen mit den Konventionsverfahren fuer Gesamtkohlenstoff [Adsorptions- (ADS) und FID-Verfahren] in Verbindung mit Einzelkomponenten-Analysen sollten die Verwendbarkeit der Gesamtkohlenstoff-Messverfahren an AVA's ueberprueft werden. Das ADS-Verfahren erwies sich dabei als nicht geeignet, da haeufig unplausibel hohe Messwerte (u.a. Stoerungen durch Querempfindlichkeiten) festgestellt wurden. Das FID-Messverfahren kann im Rahmen einer Konvention zum Einsatz kommen; da aber kein homogenes Stoffspektrum im Abgas vorliegt, ist die exakte Umrechnung der Messwerte auf den Gesamtkohlenstoff-Gehalt nicht moeglich. c) Da kein Messverfahren fuer Gesamtkohlenstoff-Vergleichsanalysen bekannt ist, sollten Hilfsverfahren fuer die Kalibrierung von FID's untersucht werden. Es wird ein Verfahren zur Bestimmung von Responsefaktoren an installierten FID's im AVA-Abgas beschrieben. d) Fuer die Bestimmung der Komponente CO an AVA's wurde das Iodpentoxid-Verfahren untersucht. Die statistische Auswertung der Messergebnisse zeigt, dass das Verfahren zur genauen Pruefgasanalyse nicht geeignet ist. Durch starke Streuungen bei Untersuchungen an realen Abgasen und die unguenstige Handhabbarkeit ist das Verfahren im Vor-Ort-Einsatz ebenfalls nicht zu empfehlen. (orig.)SIGLEAvailable from TIB Hannover: RN 8908(93-131,4) / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Umwelt, Naturschutz und Reaktorsicherheit, Bonn (Germany)DEGerman

Entwicklung eines beruehrungsfreien mikrooptischen Tonometers zur Messung des Augeninnendruckes durch Stimulation von Oberflaechenschwingungen und Messung der Bulbusvariation. Mikrooptisches Tonometer (MIOPTO) Gemeinsamer Schlussbericht

SIGLEAvailable from TIB Hannover: F03B945+a / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Bildung und Forschung, Berlin (Germany)DEGerman