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Additional file 2: Table S2. HbF and HbA2 production in ErPC cultures from 14 β-thalassemia patients after 4 and 8 days differentiation.

PNAs conjugated to carrier peptides have been employed for the targeting of miRNA precursor, with the aim to develop molecules able to interfere in the pre-miRNA processing. The capability of the molecules to bind pre-miRNA has been tested in vitro by fluorescence assayes on Thiazole Orange labeled molecules and in vivo, in K562 cells, evaluating the amount of miRNA produced after treatment of cells with two amounts of PNAs.

With this work we propose a qualitative approach, aiming to reveal every archaeological evidence of a human presence and an engraving activity in the Mount Bego region (Tende, Maritime Alps, France), during late prehistory. The typo-chronological study of carved weapons provided a chronological interval starting about 3700 B.C. and lasting until the end of the Bronze Age. Nevertheless, the discovery of archaeological material prior to this interval compels to admit the possibility that some carved patterns, different from weapons, were made in Early Neolithic, requiring consequently a diachronic interpretation of the pecked carvings set. The finding of linear schematic patterns covered by some pecked engravings denies the hypothesis that only pecked carvings date of prehistory. It is possible to consider the continuity of the engraving activity, with a coexistence of the two techniques, incised and pecked, until the end of Early Bronze Age; for the later periods it is possible to consider a progressive disuse of pecked tradition, in favour of an increasingly present schematism. Although mount Bego inscription in alpine symbolic expression is incontestable, this research revealed also the importance of the iberian cultural influences on the region rock art, from Spanish Levantine macro-schematism to linear schematic art lasting until the end of late prehistory, passing by the Bell Beaker culture through its Spanish group Ciempozuelos., Ce travail constitue une approche qualitative visant à mettre en évidence tous les indices archéologiques témoignant d'une présence humaine et d'une activité de gravure dans la région du mont Bego (Tende, Alpes-Maritimes, France) durant la préhistoire récente. L'étude typo-chronologique des gravures d'armes piquetées a fourni un intervalle chronologique débutant vers 3700 avant J.-C. et se poursuivant jusqu'à la fin de l'âge du Bronze. L'existence de matériel antérieur à cet intervalle contraint toutefois à admettre la possibilité que des motifs gravés, différents des armes, ont pu être réalisés aux débuts du Néolithique, imposant ainsi une lecture diachronique de l'ensemble piqueté de la région. La découverte de motifs dits "schématiques-linéaires" superposés par des gravures piquetées dément en outre l'hypothèse selon laquelle seules les gravures piquetées datent de la protohistoire. Une continuité dans l'activité de gravure de la région du mont Bego peut donc être envisagée, avec une coexistence des techniques incisée et piquetée jusqu'à la fin de l'âge du Bronze ancien. Pour les périodes ultérieures il est possible d'imaginer un abandon progressif de la tradition piquetée au profit d'un schématisme linéaire de plus en plus présent. Bien que l'inscription du mont Bego au sein de l'expression symbolique alpine demeure indéniable, cette étude a également permis de révéler l'importance des influences culturelles ibériques sur l'art protohistorique de la région, du macro-schématisme du Levant espagnol à l'art schématique-linéaire perdurant jusqu'à la fin de la protohistoire, en passant par la culture campaniforme au travers de son groupe espagnol Ciempozuelos.

Alessia Finotti,1–3 Laura Breda,4 Carsten W Lederer,6,7 Nicoletta Bianchi,1–3 Cristina Zuccato,1–3 Marina Kleanthous,6,7 Stefano Rivella,4,5 Roberto Gambari1–3 1Laboratory for the Development of Gene and Pharmacogenomic Therapy of Thalassaemia, Biotechnology Centre of Ferrara University, Ferrara, Italy; 2Associazione Veneta per la Lotta alla Talassemia, Rovigo, Italy; 3Department of Life Sciences and Biotechnology, Section of Biochemistry and Molecular Biology, Ferrara University, Ferrara, Italy; 4Department of Pediatrics, Division of Haematology/Oncology, Weill Cornell Medical College, New York, NY, USA; 5Department of Cell and Development Biology, Weill Cornell Medical College, New York, NY, USA; 6Department of Molecular Genetics Thalassaemia, The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus; 7Cyprus School of Molecular Medicine, Nicosia, Cyprus Abstract: The β-thalassemias are a group of hereditary hematological diseases caused by over 300 mutations of the adult β-globin gene. Together with sickle cell anemia, thalassemia syndromes are among the most impactful diseases in developing countries, in which the lack of genetic counseling and prenatal diagnosis have contributed to the maintenance of a very high frequency of these genetic diseases in the population. Gene therapy for β-thalassemia has recently seen steadily accelerating progress and has reached a crossroads in its development. Presently, data from past and ongoing clinical trials guide the design of further clinical and preclinical studies based on gene augmentation, while fundamental insights into globin switching and new technology developments have inspired the investigation of novel gene-therapy approaches. Moreover, human erythropoietic stem cells from β-thalassemia patients have been the cellular targets of choice to date whereas future gene-therapy studies might increasingly draw on induced pluripotent stem cells. Herein, we summarize the most significant developments in β-thalassemia gene therapy over the last decade, with a strong emphasis on the most recent findings, for β-thalassemia model systems; for β-, γ-, and anti-sickling β-globin gene addition and combinatorial approaches including the latest results of clinical trials; and for novel approaches, such as transgene-mediated activation of γ-globin and genome editing using designer nucleases. Keywords: Thalassemia, gene therapy, HbF induction, transcription factors, induced pluripotent stem cells, genome editing, TALEN, CRISPR, ZFN

Mouse models that carry mutations causing thalassemia represent a suitable tool to testin vivonew mutation-specific therapeutic approaches. Transgenic mice carrying theβ-globin IVSI-6 mutation (the most frequent in Middle-Eastern regions and recurrent in Italy and Greece) are, at present, not available. We report the production and characterization of a transgenic mouse line (TG-β-IVSI-6) carrying the IVSI-6 thalassemia point mutation within the humanβ-globin gene. In the TG-β-IVSI-6 mouse (a) the transgenic integration region is located in mouse chromosome 7; (b) the expression of the transgene is tissue specific; (c) as expected, normally spliced humanβ-globin mRNA is produced, giving rise toβ-globin production and formation of a human-mouse tetrameric chimeric hemoglobinαmu-globin2/βhu-globin2and, more importantly, (d) the aberrantβ-globin-IVSI-6 RNAs are present in blood cells. The TG-β-IVSI-6 mouse reproduces the molecular features of IVSI-6β-thalassemia and might be used as anin vivomodel to characterize the effects of antisense oligodeoxynucleotides targeting the cryptic sites responsible for the generation of aberrantly splicedβ-globin RNA sequences, caused by the IVSI-6 mutation. These experiments are expected to be crucial for the development of a personalized therapy forβ-thalassemia.

Effects of pre-miR-93 and antagomiR-93 on morphology and apoptosis. A. Morphology of U251 glioma cells treated for 24 h with control, pre-miR-93 and antagomiR-93 molecules (200 nM). B,C. Effects of the different treatments on apoptosis on U251 and T98G glioma cell lines, as indicated. Apoptosis was analyzed by the Annexin-V release test [60] (B) or by caspase-3/7 production [61] (C). In panel B, the effects of a positive antagomiR-221 is also shown (see Brognara et al. (2014) [62]. Data represent the average S.D. of three independent experiments. ** = p

The Vallée des Merveilles (Valley of Marvels), in the Mercantour National Park (France), is a major place for studying the late Neolithic Age until the beginning of the Bronze Age: since the early 20th century, more than 4000 engraved rocks have been listed, representing 40,000 carvings. The Laboratoire Départemental du Lazaret has been studying these engravings for more than forty years: a large database describes the whole rocks and figures and is still filled with new findings. But such a large amount of data from a vast archaeological site needs efficient tools for its study, such as mapping the location of the rocks. Many improvements regarding the study of the carvings and their spatial distribution were made possible by the development of a web GIS: consulting the data is made easier and clearer with a web-mapping platform and specific tools. Filtering for choosing the data to display and automatically generated maps allow an overall view of the spatial distribution of the Marvels’ engravings. Using both the IGN’s Geoportal data and cartographic web libraries produced an efficient and complete web service for consulting and analyzing the engraved rocks of the Valley of Marvels. This one can be used for any area in the Mercantour Park, as long as new data are added to the databases.