Conservation genetics of European wildcat (Felis silvestris silvestris): a wide and integrating analysis protocol for admixture inferences and population structure

Introgression of domestic cat genes into European wildcat (Felis silvestris silvestris) populations and reduction of wildcats’ range in Europe, leaded by habitat loss and fragmentation, are considered two of the main conservation problems for this endangered feline. This thesis addressed the questions related with the artificial hybridization and populations’ fragmentation, using a conservation genetics perspective. We combined the use of highly polymorphic loci, Bayesian statistical inferences and landscape analyses tools to investigate the origin of the geographic-genetic substructure of European wildcats (Felis silvestris silvestris) in Italy and Europe. The genetic variability of microsatellites evidenced that European wildcat populations currently distributed in Italy differentiated in, and expanded from two distinct glacial refuges during the Last Glacial Maximum. The genetic and geographic substructure detected between the eastern and western sides of the Apennine ridge, resulted by adaptation to specific ecological conditions of the Mediterranean habitats. European wildcat populations in Europe are strongly structured into 5 geographic-genetic macro clusters corresponding to: the Italian peninsular & Sicily; Balkans & north-eastern Italy; Germany eastern; central Europe; and Iberian Peninsula. Central European population might have differentiated in the extra-Mediterranean Würm ice age refuge areas (Northern Alps, Carpathians, and the Bulgarian mountain systems), while the divergence among and within the southern European populations might have resulted by the Pleistocene bio geographical framework of Europe, with three southern refugia localized in the Balkans, Italian Peninsula and Iberia Peninsula. We further combined the use of most informative autosomal SNPs with uniparental markers (mtDNA and Y-linked) for accurately detecting parental genotypes and levels of introgressive hybridization between European wild and domestic cats. A total of 11 hybrids were identified. The presence of domestic mitochondrial haplotypes shared with some wild individuals led us to hypnotize the possibility that ancient introgressive events might have occurred and that further investigation should be recommended.

Mesenchymal Stromal Cells (MSCs) and induced Plutipotent Stem Cells (iPSCs) in Domestic Animals: Characterization and Differentiation Potential

Derivation of stem cell lines from domesticated animals has been of great interest as it benefits translational medicine, clinical applications to improve human and animal health and biotechnology. The main types of stem cells studied are Embryonic Stem Cells (ESCs), induced Pluripotent Stem Cells (iPSCs) and Mesenchymal Stem/Stromal Cells (MSCs). This thesis had two main aims: (I) The isolation of bovine MSCs from amniotic fluid (AF) at different trimesters of pregnancy and their characterization to study pluripotency markers expression. Stemness markers were studied also in MSCs isolated from equine AF, Wharton’s jelly (WJ) and umbilical cord blood (UCB) as continuation of the characterization of these cells previously performed by our research group; (II) The establishment and characterization of iPSCs lines in two attractive large animal models for biomedical and biotechnology research such as the bovine and the swine, and the differentiation into the myogenic lineage of porcine iPSCs. It was observed that foetal tissues in domestic animals such as the bovine and the horse represent a source of MSCs able to differentiate into the mesodermal lineage but they do not proliferate indefinitely and they lack the expression of many pluripotency markers, making them an interesting source of cells for regenerative medicine, but not the best candidate to elucidate pluripotency networks. The protocol used to induce pluripotency in bovine fibroblasts did not work, as well as the chemical induction of pluripotency in porcine fibroblasts, while the reprogramming protocol used for porcine iPSCs was successful and the line generated was amenable to being differentiated into the myogenic lineage, demonstrating that they could be addressed into a desired lineage by genetic modification and appropriated culture conditions. Only a few cell types have been differentiated from domestic animal iPSCs to date, so the development of a reliable directed-differentiation protocol represents a very important result.

La lucha del Derecho contra el negacionismo: una peligrosa frontera. Particular estudio de los ordenamientos italiano y español

En la sociedad europea crece la preocupación por el retorno de tendencias fascistas y neonazis y por la extensión de ideologías xenófobas y antisemitas, algunas de ellas alimentadas a partir de tesis de negacionistas de aquellos trágicos eventos de nuestra historia reciente. La lucha frente a los discursos negacionistas se ha llevado más allá del ámbito social y académico, y se ha propuesto la incorporación en los ordenamientos jurídicos europeos de tipos penales específicos que incriminan este tipo de discurso: negar, banalizar, o justificar el Holocausto u otros genocidios o graves crímenes contra la humanidad. Esta legislación, que encuentra su mayor expresión en la Decisión marco 2008/913/JAI, aunque castiga un discurso socialmente repugnante, sin embargo presenta dudas en cuanto a su legitimidad con un sistema de libertades erigido sobre el pilar del pluralismo propio de los Estados democráticos. Surge así la cuestión de si pueden estar surgiendo «nuevos» delitos de opinión y a ello se dedica entonces la presente tesis. El objetivo concreto de este trabajo será analizar esta política-criminal para proponer una configuración del delito de negacionismo compatible con la libertad de expresión, aunque se cuestionará la conveniencia de castigar penalmente a través de un específico delito este tipo de conductas. En particular se pretende responder a tres preguntas: en primer lugar, ¿el discurso negacionista debe ampararse prima facie por la libertad de expresión en un ordenamiento abierto y personalista y cuáles podrían ser las «reglas» que podrían servir como criterio para limitar este género de manifestaciones? La segunda pregunta sería entonces: ¿Cómo podría construirse un tipo penal respetuoso con los principios constitucionales y penales que específicamente incriminara este género de conductas? Y, como última pregunta: ¿Es conveniente o adecuada una política criminal que lleve a crear un específico delito de negacionismo?