Multiplexed Lateral Flow Test for Detection and Differentiation of Cronobacter sakazakil Serotypes 01 and 02

The ubiquitous and opportunistic pathogen Cronobacter sakazakii is responsible for severe meningitis, sepsis, and necrotizing enterocolitis in neonates and infants associated with ingestion of contaminated powdered infant formula (PIF). The current ISO method for isolation and detection of Cronobacter spp. is laborious, time-consuming and expensive. In this study, a multiplexed lateral flow test strip was developed to rapidly detect and simultaneously serotype O1 and O2 C. sakazakil serotypes. The assay is based on two monoclonal antibodies (MAb) that specifically bind to the lipopolysaccharides (LPS) of these pathogens. The test strip provides results very quickly;C. sakazakii could be detected in pure culture within 15 min with a sensitivity of 107 CFU/ml. After non-selective enrichment for 18 h as low as one Cronobacter cell per g PIF could be detected. Moreover, the established lateral flow assay (LFA) offers excellent specificity showing no cross-reactivity with other C. sakazakii serotypes, Cronobacter species or Enterobacteriaceae tested. These characteristics, together with several advantages such as speed, simplicity in performance, low analysis cost, and no requirement of specialized skills or sophisticated equipment make the developed multiplexed LFA suitable for reliable detection and serotyping of C. sakazakii serotypes O1 and O2

The coactivator role of histone deacetylase 3 in IL-1-signaling involves deacetylation of p65 NF-κB

Histone deacetylase (HDAC) 3, as a cofactor in co-repressor complexes containing silencing mediator for retinoid or thyroid-hormone receptors (SMRT) and nuclear receptor co-repressor (N-CoR), has been shown to repress gene transcription in a variety of contexts. Here, we reveal a novel role for HDAC3 as a positive regulator of IL-1-induced gene expression. Various experimental approaches involving RNAi-mediated knockdown, conditional gene deletion or small molecule inhibitors indicate a positive role of HDAC3 for transcription of the majority of IL-1-induced human or murine genes. This effect was independent from the gene regulatory effects mediated by the broad-spectrum HDAC inhibitor trichostatin A (TSA) and thus suggests IL-1-specific functions for HDAC3. The stimulatory function of HDAC3 for inflammatory gene expression involves a mechanism that uses binding to NF-κB p65 and its deacetylation at various lysines. NF-κB p65-deficient cells stably reconstituted to express acetylation mimicking forms of p65 (p65 K/Q) had largely lost their potential to stimulate IL-1-triggered gene expression, implying that the co-activating property of HDAC3 involves the removal of inhibitory NF-κB p65 acetylations at K122, 123, 314 and 315. These data describe a novel function for HDAC3 as a co-activator in inflammatory signaling pathways and help to explain the anti-inflammatory effects frequently observed for HDAC inhibitors in (pre)clinical us

The coactivator role of histone deacetylase 3 in IL-1-signaling involves deacetylation of p65 NF-kappaB

Histone deacetylase (HDAC) 3, as a cofactor in co-repressor complexes containing silencing mediator for retinoid or thyroid-hormone receptors (SMRT) and nuclear receptor co-repressor (N-CoR), has been shown to repress gene transcription in a variety of contexts. Here, we reveal a novel role for HDAC3 as a positive regulator of IL-1-induced gene expression. Various experimental approaches involving RNAi-mediated knockdown, conditional gene deletion or small molecule inhibitors indicate a positive role of HDAC3 for transcription of the majority of IL-1-induced human or murine genes. This effect was independent from the gene regulatory effects mediated by the broad-spectrum HDAC inhibitor trichostatin A (TSA) and thus suggests IL-1-specific functions for HDAC3. The stimulatory function of HDAC3 for inflammatory gene expression involves a mechanism that uses binding to NF-?B p65 and its deacetylation at various lysines. NF-?B p65-deficient cells stably reconstituted to express acetylation mimicking forms of p65 (p65 K/Q) had largely lost their potential to stimulate IL-1-triggered gene expression, implying that the co-activating property of HDAC3 involves the removal of inhibitory NF-?B p65 acetylations at K122, 123, 314 and 315. These data describe a novel function for HDAC3 as a co-activator in inflammatory signaling pathways and help to explain the anti-inflammatory effects frequently observed for HDAC inhibitors in (pre)clinical use

Abstracts from the 8th International Conference on cGMP Generators, Effectors and Therapeutic Implications

This work was supported by a restricted research grant of Bayer AG

Regulation of NF-kappaB p65 transactivation by phosphorylation and regulatory ubiquitination

Wittwer T. Regulation of NF-kappaB p65 transactivation by phosphorylation and regulatory ubiquitination. Bielefeld; 2010

High Performance Computing in use for gravity field analysis with CHAMP, GRACE and GOCE

Das Schwerefeld der Erde ist in den letzten Jahren verstärkt zum Forschungsgegenstand geworden. Von besonderer Bedeutung zur globalen Modellierung des Schwerefeldes sind dabei Satellitenverfahren. Derzeit sind dafür die drei Satellitenmissionen CHAMP, GRACE und GOCE operierend oder in Vorbereitung. Die Auswertung dieser Daten ist aufgrund der großen Menge an Beobachtungen und Unbekannten sehr aufwändig, weshalb der Einsatz von Hochleistungsrechnern nötig ist. Diese Arbeit erläutert die Grundlagen der Schwerefeldbestimmung aus Satellitenverfahren. Detailiert werden die heute existierenden Hochleistungsrechner-Architekturen mit ihren spezifischen Vor- und Nachteilen vorgestellt. Ebenfalls eingegangen wird auf die Programmierung von Hochleistungsrechnern und die dafür geeigneten Softwarepakete. Im Rahmen dieser Arbeit wurden zwei Algorithmen zur Schwerefeldbestimmung auf verschiedenen Hochleistungsrechner-Architekturen implementiert. Die beiden Algorithmen (ein Brute-Force und ein iteratives Schätzverfahren) werden mit den Details ihrer Implementierung und Parallelisierung beschrieben. Dazu gehört auch die Einführung einer Gewichtsmatrix, die Verwendung von Regularisierung zur Stabilisierung des Gleichungssystems, sowie der Varianzkomponentenschätzung zur Kombination verschiedener Beobachtungstypen. Die Ergebnisse der Implementierung und Parallelisierung der beiden Algorithmen auf den verschiedenen verwendeten Hochleistungsrechnern werden detailiert präsentiert. Daraus lassen sich unter anderem Rückschlüsse auf die Eignung verschiedener Architekturen für die verwendeten Algorithmen ziehen. Ebenfalls vorgestellt werden die Ergebnisse der Gewichtung, der Regularisierung und der Varianzkomponentenschätzung.The earth's gravity field has increasingly been in the focus of research in recent years. Satellite methods are of special importance in the global modelling of the gravity field. For this, the three satellite missions CHAMP, GRACE and GOCE are currently operating or in preparation. The analysis of these data is, due to the large number of observations and unknowns, very complex, which requires the use of high performance computers. This work explains the basics of gravity field estimation out of satellite methods. Today's existing high performance computer architectures are presented with their respective advantages and disadvantages. Also mentioned are the programming of high performance computers and suitable software packages. In the scope of this work, two algorithms for gravity field estimation were implemented on different high performance computer architectures. The two algorithms (a brute force and an iterative estimator) are explained with the details of their implementation and parallelization. This includes the introduction of a weight matrix, the use of regularization for the stabilization of the equation system, as well as variance component estimation for the combination of different observation types. The results of the implementation and parallelization of the two algorithms on the different high performance computers used are presented in details. This allows conclusions about the suitability of various architectures for the algorithms used. Also explained are the results of weighting, regularization and variance component estimation