Customization of the lysis and reaction setup steps of FastLane Cell kits for an automated workflow

Automated workflows can be optimized for enhanced flexibility by determining the steps within the protocol that tolerate extended storage of assay components. We determined the tolerated stopping points and storage conditions in a multiplex real-time RT-PCR workflow using the FastLane Cell Multiplex Kit. Breaks were introduced within the procedure and the effect of incubation times and temperatures on the observed CT values were measured. Using these parameters, we provide guidance on adapting the protocol to enable an automated workflow. We also suggest steps for achieving optimal results when using FastLane Cell kits with TaqMan Gene Expression Assays

The prevalence of apraxia of speech in chronic aphasia after stroke.

Aphasia after stroke often comes along with a motor speech impairment called apraxia of speech (AOS). Individuals suffering from AOS can have severe problems making themselves understood, and the treatment of this problem requires specific intervention methods. Therefore, the presence or absence of speech apraxia in individuals with aphasia is of utmost clinical and therapeutic importance. Moreover, analysis of this condition is fundamental to understanding the organisation of motor speech and its neural basis within the left hemisphere language areas. While the prevalence of aphasia after stroke is relatively well studied, little is known about how many patients with aphasia have AOS. To our knowledge, there have been no systematic investigations of the prevalence of apraxic speech impairment in the aphasic population so far. This study seeks to estimate the prevalence of AOS in chronic aphasia after stroke by analysing pre-existing data from a randomized controlled multicentre trial in a sample of 156 stroke patients with aphasia of more than 6 months duration. The inclusion criteria of this earlier study replicated realistic routine care conditions in Germany. To identify patients with AOS, speech samples will be evaluated by 3 experts with more than 20 years of clinical and research experience with AOS. A prevalence estimate will be computed using a Bayesian approach

Reverse Genetic Screening Identifies Five E-class PPR Proteins Involved in RNA Editing in Mitochondria of Arabidopsis thaliana*

RNA editing in flowering plant mitochondria post-transcriptionally alters several hundred nucleotides from C to U, mostly in mRNAs. Several factors required for specific RNA-editing events in plant mitochondria and plastids have been identified, all of them PPR proteins of the PLS subclass with a C-terminal E-domain and about half also with an additional DYW domain. Based on this information, we here probe the connection between E-PPR proteins and RNA editing in plant mitochondria. We initiated a reverse genetics screen of T-DNA insertion lines in Arabidopsis thaliana and investigated 58 of the 150 E-PPR-coding genes for a function in RNA editing. Six genes were identified to be involved in mitochondrial RNA editing at specific sites. Homozygous mutants of the five genes MEF18-MEF22 display no gross disturbance in their growth or development patterns, suggesting that the editing sites affected are not crucial at least in the greenhouse. These results show that a considerable percentage of the E-PPR proteins are involved in the functional processing of site-specific RNA editing in plant mitochondria