Effects of environmental complexity on neurogenesis in the hippocampal dentate gyrus

Introduction. Postnatal neurogenesis in the hippocampal dentate gyrus, can be modulated by numerous determinants, such as hormones, transmitters and stress. Among the factors positively interfering with neurogenesis, the complexity of the environment appears to play a particularly striking role. Adult mice reared in an enriched environment produce more neurons and exhibit better performance in hippocampus-specific learning tasks. While the effects of complex environments on hippocampal neurogenesis are well documented, there is a lack of information on the effects of living under socio-sensory deprivation conditions. Due to the immaturity of rats and mice at birth, studies dealing with the effects of environmental enrichment on hippocampal neurogenesis were carried out in adult animals, i.e. during a period of relatively low rate of neurogenesis. The impact of environment is likely to be more dramatic during the first postnatal weeks, because at this time granule cell production is remarkably higher than at later phases of development. The aim of the present research was to clarify whether and to what extent isolated or enriched rearing conditions affect hippocampal neurogenesis during the early postnatal period, a time window characterized by a high rate of precursor proliferation and to elucidate the mechanisms underlying these effects. The experimental model chosen for this research was the guinea pig, a precocious rodent, which, at 4-5 days of age can be independent from maternal care. Experimental design. Animals were assigned to a standard (control), an isolated, or an enriched environment a few days after birth (P5-P6). On P14-P17 animals received one daily bromodeoxyuridine (BrdU) injection, to label dividing cells, and were sacrificed either on P18, to evaluate cell proliferation or on P45, to evaluate cell survival and differentiation. Methods. Brain sections were processed for BrdU immunhistochemistry, to quantify the new born and surviving cells. The phenotype of the surviving cells was examined by means of confocal microscopy and immunofluorescent double-labeling for BrdU and either a marker of neurons (NeuN) or a marker of astrocytes (GFAP). Apoptotic cell death was examined with the TUNEL method. Serial sections were processed for immunohistochemistry for i) vimentin, a marker of radial glial cells, ii) BDNF (brain-derived neurotrofic factor), a neurotrophin involved in neuron proliferation/survival, iii) PSA-NCAM (the polysialylated form of the neural cell adhesion molecule), a molecule associated with neuronal migration. Total granule cell number in the dentate gyrus was evaluated by stereological methods, in Nissl-stained sections. Results. Effects of isolation. In P18 isolated animals we found a reduced cell proliferation (-35%) compared to controls and a lower expression of BDNF. Though in absolute terms P45 isolated animals had less surviving cells than controls, they showed no differences in survival rate and phenotype percent distribution compared to controls. Evaluation of the absolute number of surviving cells of each phenotype showed that isolated animals had a reduced number of cells with neuronal phenotype than controls. Looking at the location of the new neurons, we found that while in control animals 76% of them had migrated to the granule cell layer, in isolated animals only 55% of the new neurons had reached this layer. Examination of radial glia cells of P18 and P45 animals by vimentin immunohistochemistry showed that in isolated animals radial glia cells were reduced in density and had less and shorter processes. Granule cell count revealed that isolated animals had less granule cells than controls (-32% at P18 and -42% at P45). Effects of enrichment. In P18 enriched animals there was an increase in cell proliferation (+26%) compared to controls and a higher expression of BDNF. Though in both groups there was a decline in the number of BrdU-positive cells by P45, enriched animals had more surviving cells (+63) and a higher survival rate than controls. No differences were found between control and enriched animals in phenotype percent distribution. Evaluation of the absolute number of cells of each phenotype showed that enriched animals had a larger number of cells of each phenotype than controls. Looking at the location of cells of each phenotype we found that enriched animals had more new neurons in the granule cell layer and more astrocytes and cells with undetermined phenotype in the hilus. Enriched animals had a higher expression of PSA-NCAM in the granule cell layer and hilus Vimentin immunohistochemistry showed that in enriched animals radial glia cells were more numerous and had more processes.. Granule cell count revealed that enriched animals had more granule cells than controls (+37% at P18 and +31% at P45). Discussion. Results show that isolation rearing reduces hippocampal cell proliferation but does not affect cell survival, while enriched rearing increases both cell proliferation and cell survival. Changes in the expression of BDNF are likely to contribute to he effects of environment on precursor cell proliferation. The reduction and increase in final number of granule neurons in isolated and enriched animals, respectively, are attributable to the effects of environment on cell proliferation and survival and not to changes in the differentiation program. As radial glia cells play a pivotal role in neuron guidance to the granule cell layer, the reduced number of radial glia cells in isolated animals and the increased number in enriched animals suggests that the size of radial glia population may change dynamically, in order to match changes in neuron production. The high PSA-NCAM expression in enriched animals may concur to favor the survival of the new neurons by facilitating their migration to the granule cell layer. Conclusions. By using a precocious rodent we could demonstrate that isolated/enriched rearing conditions, at a time window during which intense granule cell proliferation takes place, lead to a notable decrease/increase of total granule cell number. The time-course and magnitude of postnatal granule cell production in guinea pigs are more similar to the human and non-human primate condition than in rats and mice. Translation of current data to humans would imply that exposure of children to environments poor/rich of stimuli may have a notably large impact on dentate neurogenesis and, very likely, on hippocampus dependent memory functions.

New polymeric materials for vascular surgery

The dramatic impact that vascular diseases have on human life quality and expectancy nowadays is the reason why both medical and scientific communities put great effort in discovering new and effective ways to fight vascular pathologies. Among the many different treatments, endovascular surgery is a minimally-invasive technique that makes use of X-ray fluoroscopy to obtain real-time images of the patient during interventions. In this context radiopaque biomaterials, i.e. materials able to absorb X-ray radiation, play a fundamental role as they are employed both to enhance visibility of devices during interventions and to protect medical staff and patients from X-ray radiations. Organic-inorganic hybrids are materials that combine characteristics of organic polymers with those of inorganic metal oxides. These materials can be synthesized via the sol-gel process and can be easily applied as thin coatings on different kinds of substrates. Good radiopacity of organic-inorganic hybrids has been recently reported suggesting that these materials might find applications in medical fields where X-ray absorption and visibility is required. The present PhD thesis aimed at developing and characterizing new radiopaque organic-inorganic hybrid materials that can find application in the vascular surgery field as coatings for the improvement of medical devices traceability as well as for the production of X-ray shielding objects and garments. Novel organic-inorganic hybrids based on different polyesters (poly-lactic acid and poly-ε-caprolactone) and polycarbonate (poly-trimethylene carbonate) as the polymeric phase and on titanium oxide as the inorganic phase were synthesized. Study of the phase interactions in these materials allowed to demonstrate that Class II hybrids (where covalent bonds exists between the two phases) can be obtained starting from any kind of polyester or polycarbonate, without the need of polymer pre-functionalization, thanks to the occurrence of transesterification reactions operated by inorganic molecules on ester and carbonate moieties. Polyester based hybrids were successfully coated via dip coating on different kinds of textiles. Coated textiles showed improved radiopacity with respect to the plain fabric while remaining soft to the touch. The hybrid was able to coat single fibers of the yarn rather than coating the yarn as a whole. Openings between yarns were maintained and therefore fabric breathability was preserved. Such coatings are promising for the production of light-weight garments for X-ray protection of medical staff during interventional fluoroscopy, which will help preventing pathologies that stem from chronic X-ray exposure. A means to increase the protection capacity of hybrid-coated fabrics was also investigated and implemented in this thesis. By synthesizing the hybrid in the presence of a suspension of radiopaque tantalum nanoparticles, PDMS-titania hybrid materials with tunable radiopacity were developed and were successfully applied as coatings. A solution for enhancing medical device radiopacity was also successfully investigated. High metal radiopacity was associated with good mechanical and protective properties of organic-inorganic hybrids in the form of a double-layer coating. Tantalum was employed as the constituent of the first layer deposited on sample substrates by means of a sputtering technique. The second layer was composed of a hybrid whose constituents are well-known biocompatible organic and inorganic components, such as the two polymers PCL and PDMS, and titanium oxide, respectively. The metallic layer conferred to the substrate good X-ray visibility. A correlation between radiopacity and coating thickness derived during this study allows to tailor radiopacity simply by controlling the metal layer sputtering deposition time. The applied metal deposition technique also permits easy shaping of the radiopaque layer, allowing production of radiopaque markers for medical devices that can be unambiguously identified by surgeons during implantation and in subsequent radiological investigations. Synthesized PCL-titania and PDMS-titania hybrids strongly adhered to substrates and show good biocompatibility as highlighted by cytotoxicity tests. The PDMS-titania hybrid coating was also characterized by high flexibility that allows it to stand large substrate deformations without detaching nor cracking, thus being suitable for application on flexible medical devices.

La produzione drammatica dei fratelli Figueroa y Córdoba, con edizione critica di "Mentir y mudarse a un tiempo"

Diego e José de Figueroa y Córdoba furono due fratelli drammaturghi, attivi nella seconda metà del XVII secolo, che praticarono assiduamente la scrittura in collaborazione. Nel tentativo di rivalutare questi autori ingiustamente dimenticati dalla critica odierna, il presente lavoro si propone di offrire uno studio introduttivo alle figure e al teatro dei due fratelli. Nella prima parte, si offre un profilo biografico dei fratelli Figueroa, riunendo le seppur scarse notizie che possediamo sulla loro vita, in modo da inquadrarne le personalità all’interno degli ambienti letterari dell’epoca. Ci si concentra, poi, sul corpus, proponendo una panoramica delle commedie scritte da Diego e José, dando di ciascuna una breve sinossi e un rapido inquadramento all’interno dei generi tipici del teatro aureo, e segnalando gli eventuali problemi di attribuzione. Si accenna, infine, alla collaborazione dei fratelli nella scrittura di molte delle loro commedie, avanzando alcune ipotesi sulla tecnica compositiva. La seconda parte è costituita da un repertorio bibliografico del teatro “maggiore” dei due fratelli, in cui si descrivono analiticamente tutti gli esemplari conosciuti delle comedias dei Figueroa. Ogni descrizione si completa con una lista delle biblioteche in cui l’esemplare è conservato, lista compilata attraverso la consultazione di cataloghi, cartacei e on-line, delle principali biblioteche con fondi di teatro spagnolo del Siglo de Oro. Infine, la terza parte è dedicata all’edizione critica di una delle commedie scritte in collaborazione da Diego e José: Mentir y mudarse a un tiempo, condotta usando come testo base un manoscritto probabilmente autografo. L’edizione si completa di un apparato critico, con analisi dei testimoni, stemma codicum e registro delle varianti. Precede l’edizione uno studio generale introduttivo all’opera.

La narrazione spettacolare. Nuove forme dello storytelling: dai testi narrativi al teatro.

Il lavoro di tesi parte da un presupposto di osservazione empirica. Dopo aver rilevato la ricorrenza da parte di registi teatrali del panorama internazionale all’adattamento di testi non drammatici, ma narrativi per la scena, si è deciso di indagare come la natura narrativa dei testi di partenza si adatti, permanga o muti nel passaggio dal medium narrativo a quello spettacolare. La tesi si suddivide in tre parti. Nella parte intitolata “Della metodologia” (un capitolo), viene illustrato il metodo adottato per affrontare l’analisi degli spettacoli teatrali. Facendo riferimento alla disciplina semiotica teatrale, si è definito l’oggetto empirico di studio come testo spettacolare/performance text. La seconda parte della tesi, “Della comparazione dei modelli comunicativi”, (due capitoli), procede nella definizione: 1) dell’elemento teorico da indagare nei testi spettacolari, ovvero, la “narratività”; 2) dei modelli comunicativi della narrazione e dello spettacolo teatrale e delle loro intersezioni o differenze. Nella terza parte della tesi, “Della critica”, (due capitoli), vengono analizzate alcune delle opere del regista Jurij Ljubimov (Russia), Eimuntas Nekrošius (Lituania), Alvis Hermanis (Lettonia). La scelta è ricaduta sulle opere di questi registi in base a una considerazione: 1) culturale: si è deliberatamente circoscritta la sfera di indagine alla produzione teatrale russa e post-sovietica; 2) estetica: è stato osservato che la linea registica inaugurata da Jurij Ljubimov va permeando l’attività registica di registi più giovani come Nekrošius e Hermanis; 3) statistica: Ljubimov, Nekrošius, Hermanis hanno scelto di mettere in scena testi non drammatici con una elevata frequenza. La tesi è corredata da un’ampia appendice iconografica. Per l’analisi dei testi spettacolari si è fatto riferimento alla visione degli spettacoli in presa diretta in Italia e all’estero.