Quantitative sequence analysis of FBN1 premature termination codons provides evidence for incomplete NMD in leukocytes

We improved, evaluated, and used Sanger sequencing for quantification of single nucleotide polymorphism (SNP) variants in transcripts and gDNA samples. This improved assay resulted in highly reproducible relative allele frequencies (e.g., for a heterozygous gDNA 50.0+/-1.4%, and for a missense mutation-bearing transcript 46.9+/-3.7%) with a lower detection limit of 3-9%. It provided excellent accuracy and linear correlation between expected and observed relative allele frequencies. This sequencing assay, which can also be used for the quantification of copy number variations (CNVs), methylations, mosaicisms, and DNA pools, enabled us to analyze transcripts of the FBN1 gene in fibroblasts and blood samples of patients with suspected Marfan syndrome not only qualitatively but also quantitatively. We report a total of 18 novel and 19 known FBN1 sequence variants leading to a premature termination codon (PTC), 26 of which we analyzed by quantitative sequencing both at gDNA and cDNA levels. The relative amounts of PTC-containing FBN1 transcripts in fresh and PAXgene-stabilized blood samples were significantly higher (33.0+/-3.9% to 80.0+/-7.2%) than those detected in affected fibroblasts with inhibition of nonsense-mediated mRNA decay (NMD) (11.0+/-2.1% to 25.0+/-1.8%), whereas in fibroblasts without NMD inhibition no mutant alleles could be detected. These results provide evidence for incomplete NMD in leukocytes and have particular importance for RNA-based analyses not only in FBN1 but also in other genes.

Motifs slaves dans les lettres autrichiennes du XIXe siècle: tentative de compréhension et de synthèse dans le cas de Marie von Ebner-Eschenbach

Marie von Ebner-Eschenbach (1830-1916) was born and grew up in Moravia. Despite the fact that her very first language was Czech, all her literary work was written in German ; despite of her Czech origins from her fatherside, all the references to be found in her work concerning the social and national development of the Czech society of that time express, if not animosity, at least a total lack of understanding. Everthing happens as if the author just wanted to confirm and uphold the official views of the Austrian Monarchy. In this article, I’d like to show, mainly on the example of the novel Božena (1876), that a more careful reading which would take into account not only the textual statements of the writer, but as well the composition of the plot and the various behaviors of the Czech and German protagonists, could allow to bring nuances to Ebner-Eschenbach’s position towards the Czech – namely to see that she was perfectly aware and respectful of the cultural diversity and complexity of the Czech lands and that she felt a deep compassion for the claims of the minorities asking for the transformation of the Habsburg Empire into a Federation of free nations.

Poetik des Unbehagens. Unlust als 'point de départ' in Romanen Marie NDiayes

Die Romane der französischen Autorin Marie NDiaye (geb. 1967) sind durchzogen von Zuständen des Unwohlseins: NDiayes Protagonistinnen werden auf diffuse Weise sozial ausgegrenzt oder massiv bedrängt, verlieren die Kontrolle über ihren Körper oder geraten in schwindelerregende Zweifel über den Realitätsgehalt ihrer Sinneswahrnehmungen und Erinnerungen. Anhand der Romane "En famille" (1990), "Autoportrait en vert" (2005) und "Mon cœur à l’étroit" (2007) zeigt der Aufsatz, dass diese Momente des Unbehagens nicht nur die Suchbewegungen und Erkenntnisprozesse der Romanfiguren auslösen, sondern auch die kreative Spannung bilden, die den Vorgang der literarischen inventio in Gang setzt. Theoretische Modelle einer Psychologie, in der Unlust als Antrieb fungiert, finden sich bei Leibniz, Locke und Freud; Elemente einer Narratologie der Unlust, mit der sich die quête-Struktur von NDiayes Romanen beschreiben lässt, bietet die strukturalistische Märchentheorie Wladimir Propps und seiner Nachfolger.