Influence of hydrological conditions on the <Emphasis Type="Italic">Escherichia coli</Emphasis> population structure in the water of a creek on a rural watershed

Background  

Escherichia coli is a commensal bacterium of the gastro-intestinal tract of human and vertebrate animals, although the aquatic environment could be a secondary habitat. The aim of this study was to investigate the effect of hydrological conditions on the structure of the E. coli population in the water of a creek on a small rural watershed in France composed of pasture and with human occupation.     

Combined MRI and 31P-MRS Investigations of the ACTA1(H40Y) Mouse Model of Nemaline Myopathy Show Impaired Muscle Function and Altered Energy Metabolism

Nemaline myopathy (NM) is the most common disease entity among non-dystrophic skeletal muscle congenital diseases. Mutations in the skeletal muscle α-actin gene (ACTA1) account for ∼25% of all NM cases and are the most frequent cause of severe forms of NM. So far, the mechanisms underlying muscle weakness in NM patients remain unclear. Additionally, recent Magnetic Resonance Imaging (MRI) studies reported a progressive fatty infiltration of skeletal muscle with a specific muscle involvement in patients with ACTA1 mutations. We investigated strictly noninvasively the gastrocnemius muscle function of a mouse model carrying a mutation in the ACTA1 gene (H40Y). Skeletal muscle anatomy (hindlimb muscles and fat volumes) and energy metabolism were studied using MRI and ³¹Phosphorus magnetic resonance spectroscopy. Skeletal muscle contractile performance was investigated while applying a force-frequency protocol (from 1–150 Hz) and a fatigue protocol (80 stimuli at 40 Hz). H40Y mice showed a reduction of both absolute (−40%) and specific (−25%) maximal force production as compared to controls. Interestingly, muscle weakness was associated with an improved resistance to fatigue (+40%) and an increased energy cost. On the contrary, the force frequency relationship was not modified in H40Y mice and the extent of fatty infiltration was minor and not different from the WT group. We concluded that the H40Y mouse model does not reproduce human MRI findings but shows a severe muscle weakness which might be related to an alteration of intrinsic muscular properties. The increased energy cost in H40Y mice might be related to either an impaired mitochondrial function or an alteration at the cross-bridges level. Overall, we provided a unique set of anatomic, metabolic and functional biomarkers that might be relevant for monitoring the progression of NM disease but also for assessing the efficacy of potential therapeutic interventions at a preclinical level.     

EEG Investigations of Duration Discrimination: The Intermodal Effect Is Induced by an Attentional Bias

Previous studies indicated that empty time intervals are better discriminated in the auditory than in the visual modality, and when delimited by signals delivered from the same (intramodal intervals) rather than from different sensory modalities (intermodal intervals). The present electrophysiological study was conducted to determine the mechanisms which modulated the performances in inter- and intramodal conditions. Participants were asked to categorise as short or long empty intervals marked by auditory (A) and/or visual (V) signals (intramodal intervals: AA, VV; intermodal intervals: AV, VA). Behavioural data revealed that the performances were higher for the AA intervals than for the three other intervals and lower for inter- compared to intramodal intervals. Electrophysiological results indicated that the CNV amplitude recorded at fronto-central electrodes increased significantly until the end of the presentation of the long intervals in the AA conditions, while no significant change in the time course of this component was observed for the other three modalities of presentation. They also indicated that the N1 and P2 amplitudes recorded after the presentation of the signals which delimited the beginning of the intervals were higher for the inter- (AV/VA) compared to the intramodal intervals (AA/VV). The time course of the CNV revealed that the high performances observed with AA intervals would be related to the effectiveness of the neural mechanisms underlying the processing of the ongoing interval. The greater amplitude of the N1 and P2 components during the intermodal intervals suggests that the weak performances observed in these conditions would be caused by an attentional bias induced by the cognitive load and the necessity to switch between modalities.     

Beitrag zur Kenntnis der chemischen Zusammensetzung derPhragmiti-Magnocaricetea und derAgropyro-Rumicion-Arten

Zwanzig Pflanzen der KlassePhragmiti-Magnocaricetea (Kennarten der einzelnen pflanzen-soziologischen Einheiten) und des VerbandesAgropyro-Rumicion crispi wurden den chemischen Analysen unterzogen. Die Pflanzenproben wurden auf den Wiesen der Trockengebiete S-Mährens und der SW-Slowakei gesammelt, u. zw. aus den pflanzensoziologisch, definierbaren Einheiten (Assoziationen der VerbändePhragmition, Caricion gracilis, Caricion rostratae undAgropyro-Rumicion). Die Resultate wurden mit der chemischen Zusammensetzung der in anderen Gebieten vorkommenden Pflanzen verglichen. Es zeigten sich Zusammenhänge zwischen der genetisch bedingten chemischen Zusammensetzung bestimmter Artengruppen und den Eigenschaften des Substrats. In diesem Sinne gibt es allgemeine Unterschiede zwischen den VerbändenCaricion gracilis undCaricion rostratae. Die Arten desAgropyro-Rumicion zeigen im Durchschnitt engere Bindung an die Gesellschaften desCnidion-Verbandes als an dasCaricion gracilis.     

The SSV-Seq 2.0 PCR-Free Method Improves the Sequencing of Adeno-Associated Viral Vector Genomes Containing GC-Rich Regions and Homopolymers

Adeno-associated viral vectors (AAV) are efficient engineered tools for delivering genetic material into host cells. The commercialization of AAV-based drugs must be accompanied by the development of appropriate quality control (QC) assays. Given the potential risk of co-transfer of oncogenic or immunogenic sequences with therapeutic vectors, accurate methods to assess the level of residual DNA in AAV vector stocks are particularly important. An assay based on high-throughput sequencing (HTS) to identify and quantify DNA species in recombinant AAV batches is developed. Here, it is shown that PCR amplification of regions that have a local GC content >90% and include successive mononucleotide stretches, such as the CAG promoter, can introduce bias during DNA library preparation, leading to drops in sequencing coverage. To circumvent this problem, SSV-Seq 2.0, a PCR-free protocol for sequencing AAV vector genomes containing such sequences, is developed. The PCR-free protocol improves the evenness of the rAAV genome coverage and consequently leads to a more accurate relative quantification of residual DNA. HTS-based assays provide a more comprehensive assessment of DNA impurities and AAV vector genome integrity than conventional QC tests based on real-time PCR and are useful methods to improve the safety and efficacy of these viral vectors.