2,241 research outputs found
Exon Skipping Quantification by Real-Time PCR
No full textAntisense oligonucleotide (AON)-mediated exon skipping is a therapeutic approach for subsets of Duchenne muscular dystrophy (DMD) patients to ameliorate the severe DMD phenotype. Several groups have successfully induced exon skipping by AONs to reframe the mRNA in various patients carrying deletions, and phase I/II clinical trials are ongoing. The approach is based on targeting specific splicing motifs, both exonic and located on the exon borders, thus interfering with the spliceosome assembly by steric hindrance. Evaluation of the effectiveness of treatment with AONs in cells, animal models, and humans requires a sensitive, specific, and highly reproducible method. We have developed a real-time PCR-based protocol that uses the probe-based approach to recognize specific sequences internal to the target exon (exon-specific real-time assay). The methods for this protocol are described in this chapter. © 2012 Springer Science+Business Media, LLC
A shape to fit the need: how cell rearrange their organelle composition and architecture during development and stress (PRIN 2017, codice 2017XA5J5N)
No full textOur proposal investigates how cells reshape their organelles during development towards a secretory specialization, focusing on the mitochondria-endoplasmic reticulum (ER) contacts. It starts from the assumption that a higher ATP demand imposed by increased ER protein folding and degradation could be satisfied by formation of functional contacts with mitochondria. Revisiting mitochondrial-associated membranes (MAM) biogenesis under dynamic conditions will allow identifying molecules and patterns induced to satisfy the increased energetic demand. To this end, we will deploy state of the art quantitative proteomics and imaging technologies to analyze cellular systems induced to synthesize abundant secretory proteins with different fates, recently developed and characterized in our labs (Bakunts et al., 2017). Do MAM increase/change in parallel to the expansion of the secretory pathway? Does an increased ATP demand in the ER induce more/tighter MAM? Do different cargoes impact the outcome, and if so how? Does the unfolded protein response drive MAM reshaping, and if so via which branch(es)? Our experiments will shed light on fundamental functions of eukaryotic cells with relevance for various diseases that originate from altered ER and mitochondrial functions
In vivo study of an aberrant dystrophin exon inclusion in X-linked dilated cardiomyopathy
No full textWe previously identified a dystrophin intron 11 rearrangement in one family with X-linked dilated cardiomyopathy, causing incorporation of an aberrant exon in a tissue-specific manner. In this study we analyzed the role of different intron 11 genomic regions in the regulation of splicing by using mini-genes based approach, in C2C12 (skeletal muscle) myoblasts and myotubes, H9C2 cardiomyocytes, and HeLa cells. We show that inclusion of the aberrant exon is favored in H9C2 and differentiated C2C12 myotubes. These data suggest that the aberrant exon undergoes a differentiation-specific splicing. Unexpectedly, length of intron has a favorable effect in inclusion of the aberrant exon in the cardiac cells, suggesting that cardiac cells might be more prone to steric hindrance of trans-acting factors, involved in the inclusion of the aberrant exon. Furthermore, the cultured cell system used can serve as a suitable model to study human alternative splicing
Filologia editoriale, Roberto Calasso in dialogo con Paola Italia e Francisco Rico
No full textPaola Italia e Francisco Rico intervengono sul libro di Roberto Calasso, presidente e fondatore di Adelphi Edizioni, L'impronta dell'editore, e discutono di problemi di filologia delle forme editoriali, dal punto di vista dell'autore, del lettore e dell'editore.Paola Italia and Francisco Ricos interview Roberto Calasso, Publisher, Writer, and Founder of Adelphi Edizioni, about his book: L'impronta dell'editore, talking about philology, publishing and editing, from the author, the reader and the publisher's point of view
Paola Gianturco: Women Who Light the Dark
No full textPaola Gianturco is a photographer, author, and advocate for women\u27s rights world-wide. For the past thirteen years, she has worked as a photojournalist, documenting women’s lives in forty countries. She has published four acclaimed photo books which bring together inspiring stories with gorgeous photographs to motivate her readers to engage with, learn from and support women around the world. All of Gianturco’s books are philanthropic projects, for which she donates her royalties to carefully selected nonprofit organizations that relate to each book\u27s content. Paola\u27s most recent book, Women Who Light the Dark, tells the story of local women around the world who are helping one another tackle the problems that darken their lives—including violence, poverty, illiteracy and disease. Gianturco is giving 100% of her author royalties for this book to The Global Fund for Women.https://thekeep.eiu.edu/humanitiescenter_meaningfulwork1011/1002/thumbnail.jp
Antisense oligonucleotides capable of inducing exon skipping in dystrophin gene and their use in treatment of duchenne muscular dystrophy
No full textThe invention relates to antisense oligonucleotides capable of inducing exon skipping in dystrophin gene and their use in treatment of Duchenne muscular dystrophy (DMD). Skipping of said exons from the mRNA dystrophin transcript promotes prodn. of a functional or partially functional dystrophin protein in a patient carrying a small mutation in said exon. Such a use is characterized in that the exon is selected from the group consisting of exon 10, exon 16, exon 26, exon 33 and exon 34 of the dystrophin gene
«Yo soy de Destierrolandia»: Fernando Arrabal y el límite
No full textFernando Arrabal is a Spanish author - actually, no, he isn't. He is a francophone writer, or, more accurately, a Spanish expatriate. Or, on the contrary, he is a traitor, an «afrancesado». In this paper Fernando Arrabal's case will be used as an example of the difficulties that an author has to face when he leaves his own country and moves to another different from a spatial, cultural, and linguistic point of view. Besides, I will analyze the unusual position of the critic in the reception and interpretation of a corpus that, while being obviously connected to its author, cannot refer to a precise national context
Introduzione, a Mario Tobino, Il Clandestino
No full textL'introduzione presenta il libro più ambizioso di Mario Tobino, Il Clandestino, dedicato al racconto della sua esperienza con i gruppi clandestini della Resistenza Viareggina, libro con cui l'autore vinse il Premio Strega, nel 1962, imponendosi all'attenzione del grande pubblico dopo il successo dei libri manicomiali. Una nuova edizione in cui Paola Italia valorizza i materiali inediti dell'Archivio Tobino conservato presso l'Archivio Contemporaneo A. Bonsanti del Gabinetto GP Vieusseux di Firenze,Paola Italia presents an Introduction to the new edtion of Il Clandestino, the most ambitious of Mario Tobino's novels, dedicated to the story of his experience with the groups of the Tuscan Resistance when he was a psichiatric doctor ar Lucca. With this book the author won the Strega Prize, in 1962, attracting the attention of a wide public after the success of his psichiatric books. A new edition where Paola Italia enhances the unpublished material of Tobino Archive preserved in A. Bonsanti Contemporary Archive of G.P. Vieusseux Cabinet of Florence
Nanoparticle of the core-shell type suitable for delivering therapeutic oligonucleotides to target tissues and the use thereof for the preparation of a medicament for treating duchenne muscular distrophy
No full textA nanoparticle of the core-shell type is described, which is suitable for delivering therapeutic oligonucleotides into muscle fibers, particularly antisense oligoribonucleotides capable of restoring, in all or in part, the expression of the dystrophin protein by the exon skipping mechanism, and thus effective in the treatment of Duchenne muscular dystrophy. Moreover, a nanoparticle/oligonucleotide complex effective in the treatment of Duchenne muscular dystrophy is described
Tempo and mode of evolution of a primate-specific retrotransposon belonging to the LINE 1 family.
No full textL1P_MA2 is a primate-specific subfamily of L1 retrotransposons. The consensus sequence of this element differs from the canonical L1 consensus by the presence of a 3800-bp region in 5' (L1M1_5). Part of this region has been proposed to be involved in a dystrophin mutation affecting the correct splicing of the gene and causing an X-linked dilated cardiomyopathy. In consideration of the potential involvement in splicing regulation of this element and also because of its atypical structure, we investigated its evolutionary history by analyzing the inter- and intraspecific divergence of L1P_MA2 sequences in various species of primates. The resulting phylogenetic trees show long terminal branches and short basal internodes, as expected for a rapid event of diversification that occurred in the past. The phylogenetic analysis and the intraspecific divergence estimates revealed a pattern of evolution for this element similar in all primates with the exception of lemurs, thus suggesting that the major wave of expansion of L1P_MA2 in primate genomes occurred after the divergence between Prosimiae and Anthropoidea. These results clearly indicate that a phylogenetic approach is more appropriate than methods based on sequence data from a single species, when investigating time and mode of evolution of retro-elements
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