I linfomi a cellule T periferiche rappresentano circa il 12% di tutte le neoplasie linfoidi.In questo studio, abbiamo effettuato un’analisi di miRNA profiling (TaqMan Array MicroRNA Cards A) su 60 campioni FFPE suddivisi in: PTCLs/NOS (N=25), AITLs (N=10), ALCLs (N=12) e cellule T normali (N=13). Abbiamo identificato 4 miRNA differenzialmente espressi tra PTCLs e cellule T normali. Inoltre, abbiamo identificato tre set di mirna che discriminano le tre entita di PTCLs nodali / AIMs: Here, we performed an extensive miRNA profiling of PTCLs in order to identify differentially expressed miRNA, either involved in their pathogenesis or potentially useful for their differential diagnosis. Methods: We studied by miRNA profiling (TaqMan Array MicroRNA Cards A) 60 samples including PTCLs/NOS (N=25), AITLs (N=10), ALCLs (N=12) and normal T cells (N=13); in addition, 40 independent PTCL cases were studied by qRT-PCR as validation. We assess a GEP and miRNA Profiling. Findings: we identified 256 miRNA differentiating the two groups. In conclusion, miRNA profiling allowed to identify miRNA possibly involved in PTCL pathogenesis and to develop a novel diagnostic tool for the differential diagnosis of the commonest subtypes.

Traditional morphological examinations are not anymore sufficient for a complete evaluation of tumoral tissue and the use of neoplastic markers is of utmost importance. Neoplastic markers can be classified in: diagnostic, prognostic and predictive markers. Three markers were analyzed. 1) Insulin-like growth factor binding protein 2 (IGFBP2) was immunohistochemically examined in prostatic tissues: 40 radical prostatectomies from hormonally untreated patients with their preoperative biopsies, 10 radical prostatectomies from patients under complete androgen ablation before surgery and 10 simple prostatectomies from patients with bladder outlet obstruction. Results were compared with α-methylacyl-CoA racemase (AMACR). IGFBP2 was expressed in the cytoplasm of untreated adenocarcinomas and, to a lesser extent, in HG-PIN; the expression was markedly lower in patients after complete androgen ablation. AMACR was similarly expressed in both adenocarcinoma and HG-PIN, the level being similar in both lesions; the expression was slightly lower in patients after complete androgen ablation. IGFBP2 may be used a diagnostic marker of prostatic adenocarcinomas. 2) Heparan surface proteoglycan immunohistochemical expression was examined in 150 oral squamous cell carcinomas. Follow up information was available in 93 patients (range: 6-34 months, mean: 19±7). After surgery, chemotherapy was performed in 8 patients and radiotherapy in 61 patients. Multivariate and univariate overall survival analyses showed that high expression of syndecan-1 (SYN-1) was associated with a poor prognosis. In patients treated with radiotherapy, such association was higher. SYN-1 is a prognostic marker in oral squamous cell carcinomas; it may also represent a predictive factor for responsiveness to radiotherapy. 3) EGFR was studied in 33 pulmonary adenocarcinomas with traditional DNA sequencing methods and with two mutation-specific antibodies. Overall, the two antibodies had 61.1% sensitivity and 100% specificity in detecting EGFR mutations. EGFR mutation-specific antibodies may represent a predictive marker to identify patients candidate to tyrosine kinase inhibitors therapy.

Esta tesis doctoral propone valorar objetivamente el estímulo al realizar la técnica osteopática “rodamiento alternativo de temporales”, establecer y validar el protocolo más eficaz de esta técnica en el descenso de la presión arterial (PA) y de la frecuencia cardiaca (FC), estudiando su efecto inmediato y a lo largo de un mes. Hemos observado que la valoración subjetiva del osteópata al realizar la técnica se corresponde con los datos de tiempo obtenidos a través del método empleado. El protocolo más eficaz de la técnica es realizarla una vez a la semana, con apoyo en zona occipital y en apófisis mastoides. Su efecto no depende de quien la realice ni de la fuerza utilizada. La realización de la técnica consigue un descenso de la PA y de la FC de forma inmediata y durante 4 semanas. Esta técnica ha demostrado su utilidad como coadyuvante al tratamiento farmacológico de la hipertensión arterial. / This doctoral thesis proposes to assess objectively the induced stimulus for the “Alternative Rocking of Temporal Bones” Osteopathic Technique, establish and validate the most efficient procedure of this technique on the decrease of Blood Pressure (BP) and Heart Rate (HR), evaluating its immediate effect and throughout a month. The subjective assessment of the Osteopath during the technique procedure has been matched up with the obtained time data by the technique executed. Performing the technique leaning in occipital and mastoid bones once a week, is the most efficient procedure. No matter who performs the technique or the applied strength to achieve its effect. This technique achieves a decrease of BP and HR immediately and during the following 4 weeks. This technique has proved useful as an adjunct to pharmacological treatment of hypertension.

During my PhD,I have been develop an innovative technique to reproduce in vitro the 3D thymic microenvironment, to be used for growth and differentiation of thymocytes, and possible transplantation replacement in conditions of depressed thymic immune regulation. The work has been developed in the laboratory of Tissue Engineering at the University Hospital in Basel, Switzerland, under the tutorship of Prof.Ivan Martin. Since a number of studies have suggested that the 3D structure of the thymic microenvironment might play a key role in regulating the survival and functional competence of thymocytes, I’ve focused my effort on the isolation and purification of the extracellular matrix of the mouse thymus. Specifically, based on the assumption that TEC can favour the differentiation of pre-T lymphocytes, I’ve developed a specific decellularization protocol to obtain the intact, DNA-free extracellular matrix of the adult mouse thymus. Two different protocols satisfied the main characteristics of a decellularized matrix, according to qualitative and quantitative assays. In particular, the quantity of DNA was less than 10% in absolute value, no positive staining for cells was found and the 3D structure and composition of the ECM were maintained. In addition, I was able to prove that the decellularized matrixes were not cytotoxic for the cells themselves, and were able to increase expression of MHC II antigens compared to control cells grown in standard conditions. I was able to prove that TECs grow and proliferate up to ten days on top the decellularized matrix. After a complete characterization of the culture system, these innovative natural scaffolds could be used to improve the standard culture conditions of TEC, to study in vitro the action of different factors on their differentiation genes, and to test the ability of TECs to induce in vitro maturation of seeded T lymphocytes.

El Diagnòstic Genètic Preimplantacional (o DGP) és un conjunt de metodologies, que s'apliquen en el transcurs d'un cicle de reproducció assistida, i que permeten la selecció d'embrions sans en parelles afectes de malalties genètiques hereditàries com la fibrosi quística o l'hemofília, entre moltes altres.A més, també s'utilitza per realitzar un cribatge d'alteracions cromosòmiques (o aneuploïdies) que puguin afectar l'embrió, ja que s'ha evidenciat que aquestes són molt abundants en avortaments. S'ha postulat que una selecció positiva dels embrions lliures de aneuploïdies (és a dir, euploides) hauria d'augmentar la taxa d'embaràs dels embrions transferits al pacient. Publicacions recents, però, han demostrat que encara que es seleccionin els embrions euploides, la taxa d'embaràs no millora significativament, essent al voltant del 13%, mentre que en pacients als quals no s'aplica el DGP és del 30%. Un dels motius per aquest reduït èxit bé pot ser la tècnica utilitzada per realitzar el cribatge de aneuploïdies, la FISH, que limita l'estudi a només nou dels 23 cromosomes de l'embrió, quedant doncs més de la meitat sense diagnosticar. Per tant, els embrions que es transfereixen com euploides utilitzant la FISH poden contenir aneuploïdies per a qualsevol dels cromosomes restants no analitzats, fet que implica que l'embrió no implanti o que no generi un embaràs viable. Alternativament a la FISH, existeix la CGH que permet la detecció de tots els 23 cromosomes de l'embrió. La CGH s'ha aplicat prèviament en el DGP, però té l'inconvenient del temps necessari per realitzar la metodologia, que és de tres dies i que implica que no hi ha prou temps per diagnosticar els embrions abans de transferir-los a la pacient. Així doncs, els embrions s'han de congelar a l'espera d'obtenir els resultats de la CGH, per a ser transferits els que siguin euploides en un altre cicle de reproducció assistida. Aquest procés té apart de la desavantatge de les molèsties que implica la necessitat d'aplicar dos cicles de reproducció assistida contigus a la pacient, el fet que entre el 20-40% dels embrions no sobreviuen al procés de congelació / descongelació. Això pot suposar que encara que un embrió hagi estat diagnosticat com euploide per a tots els cromosomes mitjançant CGH, no es pugui arribar a transferir degut a no sobreviure a la congelació / descongelació. Per tal de solucionar aquestes desavantatges, una proposta metodològica alternativa és analitzar indirectament el ovòcit mitjançant estudi del corresponent 1er corpuscle polar (1CP). Amb aquesta alternativa, consistent en biopsiar el 1CP a després de la fecundació (o Dia 0), es disposa de fins a 4 dies per obtenir el resultat de la CGH. Això possibilita poder transferir en el mateix cicle de reproducció assistida els embrions derivats d'oòcits potencialment euploides, sense necessitat de congelar i descongelar. Aquesta proposta té el desavantatge, de quedar fora d'anàlisi les anomalies cromosòmiques d'origen masculí o les produïdes en el propi embrió durant les primeres divisions. Malgrat aquesta limitació, i atès que el 80% de les aneuploïdies l'embrió s'originen en l'ovòcit, l'anàlisi amb CGH de l'ovòcit permet detectar la majoria d'embrions aneuploides. Per això aquesta aproximació metodològica és molt adequada per al DGP. En aquesta tesi doctoral, s'ha aplicat un protocol de DGP per evitar malalties genètiques conjuntament amb un cribatge de aneuploïdies d'origen femení mitjançant la CGH aplicada al 1CP. Aquest doble diagnòstic genètic s'ha definit com diagnòstic genètic preimplantacional de doble factor (o DF-PGD) i ha estat aplicat, durant aquesta tesi doctoral per primera vegada en tot el món. Concretament, el DF-PGD s'ha aplicat a embrions de deu famílies afectes de Fibrosi Quística, la Síndrome d'Angelman o bé de Von Hippel-Lindau, aconseguint incrementar la taxa d'embaràs fins al 33%, respecte del 12,5% que és la que s'obté quan no es realitza aquest doble anàlisi. S'ha produït el naixement de quatre nadons sans per les respectives malalties familiars. Tot i admetent que aquests resultats són preliminars, apunten que el procediment del DF-PGD és útil per diagnosticar les malalties genètiques hereditàries i a la vegada, permet un augment considerable de la taxa d'embaràs en aquest grup de pacients. Amb l'objectiu de constatar si el DF-PGD indicat no només a dones d'edat avançada, sinó també en dones joves, aquesta tesi inclou un estudi citogenètic complet d'ovòcits de dones joves, analitzant mitjançant la CGH ambdues cèl·lules constituent del ovòcit madur: el 1CP i la corresponent MII. S'han inclòs en l'estudi dels 84 oòcits de dones joves (de 24 a 26 anys) que participen en programes de donació d'ovòcits en centres de reproducció assistida. S'ha evidenciat que un 40% dels ovòcits contenen aneuploïdies. Així doncs, el DF-PGD pot ser una bona opció per millorar l'actual taxa d'embaràs en pacients amb malalties genètiques hereditàries, independentment de l'edat de la pacient. En conclusió, en aquesta tesi doctoral s'ha evidenciat que el DF-PGD és una variant de DGP que no només és molt recomanada per a parelles en les quals la dona té una edat reproductiva avançada (més de 35 anys) sinó també per a parelles joves, i que pot resultar molt útil per incrementar les actuals taxa d'embaràs. / Preimplantation genetic diagnosis (or PGD) is a set of methodologies that is applied during a cycle of assisted reproduction, allowing the selection of healthy embryos in couples suffering from inherited genetic diseases as cystic fibrosis, haemophilia and many others. It also is used to perform a screening for chromosomal abnormalities (or aneuploidy) that may affect the embryo, as it has become clear that these are very abundant in abortions. It has been postulated that a positive selection of embryos free of aneuploidy (i.e. euploid embryos) should increase the pregnancy rate of embryos transferred to the patient. Recent publications, however, have shown that although the selection of euploid embryos, the pregnancy rate does not improve significantly, with about 13%, whereas patients who did not apply the DGP are 30%. One reason for this limited success may well be the technique used for screening for aneuploidy, the FISH, which limits the study to only nine of the 23 chromosomes of the embryo, thus leaving more than half undiagnosed. Therefore, the embryos are transferred as euploid using FISH may contain aneuploidy of the other chromosomes not analyzed, which implies that either a failure of the embryo implantation.Alternatively to the FISH, the CGH technique allows the detection of all 23 chromosomes of the embryo. The CGH has been applied previously in the PGD, but has the disadvantage of the time required for the methodology, which is three days and that means that there is not enough time to diagnose embryos before transferring them to the patient. Thus, embryos must be frozen to await the results of the CGH to be transferred. This process has the disadvantage of being apart from the discomfort that implies the need to implement two cycles of assisted reproduction to the patient, the fact that between 20-40% of embryos do not survive the process of freezing / thawing. This may mean that while an embryo has been diagnosed as euploide for all chromosomes by CGH, can not be transferred due to not surviving the freezing / thawing process.To overcome these disadvantages, an alternative methodology is to analyze indirectly through study of the corresponding oocyte corpuscle 1st Polar (1PB). With this alternative, consisting of the biopsy in 1PB after fertilization (or Day 0), it is up to 4 days for the result of the CGH. This enables transfer in the same cycle of assisted reproduction embryos derived from oocytes euploides potentially without the need to freeze and thaw. This proposal has the disadvantage of being left out of the analysis of chromosomal abnormalities or male origins were produced in the embryo during the first division. Despite this limitation, given that 80% of embryo aneuploidy originates in the oocyte, the CGH analysis of oocytes can detect the majority of aneuploid embryos. Therefore the methodological approach is very suitable for the DGP. In this thesis, we have implemented a protocol for PGD to avoid disease with a genetic screening for aneuploidy in female origin by CGH applied to 1PB. This dual genetic diagnosis has been defined as preimplantation genetic diagnosis of double-factor (or DF-PGD) and has been applied during this thesis for the first time around the world. Specifically, the DF-PGD has been applied to embryos of ten families affected by cystic fibrosis, Angelman syndrome or von Hippel-Lindau, achieving pregnancy rate increased to 33% on the 12.5% which is obtained when the DF-PGD is not analysis. There was the birth of four healthy babies by their illness relatives. Even accepting that these results are preliminary, it suggests that the proceedings of the DF-PGD is useful for diagnosing hereditary genetic diseases and also allows an increase in pregnancy rate in this group of patients. Aiming to establish whether the DF-PGD indicated not only older women but also young women, this thesis includes a comprehensive cytogenetic study of oocytes from young women looking through CGH both constituent cells of mature oocyte, the 1CP and the corresponding MII. We performed a study of young women 84 oocytes (24 to 26 years) involved in egg donation programs in assisted reproduction centres. After its analysis, we concluded that 40% of the oocytes contain aneuploidy. Thus, the DF-PGD may be a good option to improve the current pregnancy rate in patients with inherited genetic diseases, regardless of the age of the patient.In conclusion, this thesis has shown that the DF-PGD is a variant of PGD is not only highly recommended for couples where the woman has an advanced age (over 35 years) but also for young couples and that can be useful to increase the current rate of pregnancy.

Les lesions nervioses han estat des de sempre un gran repte per al cirurgià. Si a més, van associades a defectes cutanis, la situació és devastadora. En aquests casos, molts estudis demostren que s’obtenen millors resultats reconstruint el nervi amb empelts nerviosos vascularitzats, cobrint el defecte cutani amb un penjall. En la literatura es troben descrits pocs penjalls compostos de nervi i pell per aquestes situacions. L’objectiu del nostre treball ha estat dissenyar tres penjalls neurocutanis compostos pel nervi cubital i la pell medial del braç, el nervi cutani avantbraquial lateral i la pell lateral del terç distal del braç i el terç proximal de l’avantbraç, i finalment la branca sensitiva del nervi radial i la pell lateral de l’avantbraç, incloent en aquest darrer cas el múscul braquiorradial com a transferència muscular motoritzada. Per açò hem dissecat 45 extremitats superiors criopreservades i injectades amb làtex negre, distribuïdes en tres grups de 15 espècimens en cada un. S’han recollit les dades dels vasos d’origen de cada un dels nervi i de les perforants de la pell associada. En cas del nervi radial, també s’ha estudiat l’origen de la vascularització del múscul braquiorradial. Posteriorment hem valorat si existeix un origen comú de la irrigació d’aquestes estructures per determinar si és possible la realització segura de penjalls compostos neurocutanis o neuromusculocutanis. Els resultats obtinguts confirmen l’existència d’un origen comú de la vascularització de les estructures estudiades en cada un dels treballs. D’aquesta manera, hem vist que el nervi cubital i la pell medial del braç estan irrigats per l’artèria col•lateral cubital superior que permet la utilització d’aquest penjall de forma lliure. La vascularització del nervi cutani avantbraquial lateral i la pell lateral del terç distal del braç i el terç proximal de l’avantbraç prové de branques de l’artèria radial o l’artèria recurrent radial. Açò permet l’ús d’aquest penjall tant de forma lliure com pediculada. Finalment hem observat que la branca sensitiva del nervi radial, el múscul braquiorradial i la pell lateral de l’avantbraç es vascularitzen per branques de l’artèria radial amb una important contribució de l’artèria recurrent radial. Així, també és factible el disseny d’un penjall compost tant pediculat com lliure. / Nerve lesions have always been a great challenge for surgeon. When they are associated to cutaneous defect, the situation is even more devastating. In those cases, many studies demonstrate better results reconstructing the nerve with a vascularized nerve graft, using a skin flap to solve the coverage defect. There are few composted flaps of nerve and skin described in the literature. Our objective was to design three neurocutaneous flaps composed by the ulnar nerve and the medial arm skin, the lateral antebrachial cutaneous nerve and the lateral skin of the distal third of the arm and the proximal third of the forearm, and finally the sensitive branch of the radial nerve and the lateral forearm skin, including in this case the brachioradialis muscle. We dissected 45 injected cryopreserved upper extremities which were distributed into three groups of 15 specimens each one. Data about the origin of the vascularization of each nerve and the source of the perforator vessels to the associated skin were registered. Afterwards, we evaluated whether the vascularization of these structures come from the same arteries or not, in order to determine if the realization of composed neurocutaneous or neuromusculocutaneous flaps is feasible or not. Our results confirm the existence of a common source of the vascularization of the studied structures in each work. So, we found that the ulnar nerve and the medial arm skin are irrigated by the superior ulnar collateral artery which allow the use of this flap either pediculated or free. The vascularization of the lateral antebrachial cutaneous nerve and the lateral skin of the distal third of the arm and the proximal third of the forearm comes from branches of the radial artery and the radial recurrent artery. It also permits the use of this flap either free or pediculated. Finally, we found that the sensitive branch of the radial nerve and the lateral forearm skin are irrigated by the radial artery, being the radial recurrent artery important as well. So, it is possible to design a composted flap of both structures either pediculated or free.

Background. Neoangiogenesis is crucial in plaque progression and instability. Previous data from our group demonstrated that intra-plaque neovessels show both a Nestin+/WT+ and a Nestin+/WT1- phenotype, the latter being correlated with complications and plaque instability. Aims. The aims of the present thesis are: (i) to confirm our previous results on Nestin/WT1 phenotype in a larger series of carotid atheromatous plaques, (ii) to evaluate the relationship between the Nestin+/WT1- neoangiogenesis phenotype and plaque morphology, (iii) to evaluate the relationship between the immunohistochemical and histopathological characteristics and the clinical instability of the plaques. Materials and Methods. Seventy-three patients (53 males, 20 females, mean age 71 years) were consecutively enrolled. Symptoms, brain CT scan, 14 histological variables, including intraplaque hemorrhage and diffuse calcifications, were collected. Immunohistochemistry for CD34, Nestin and WT1 was performed. RT-PCR was performed to evaluate Nestin and WT1 mRNA (including 5 healthy arteries as controls). Results. Diffusely calcified plaques (13 out of 73) were found predominantly in females (P=0.017), with a significantly lower incidence of symptoms (TIA/stroke) and brain focal lesions (P=0.019 and P=0.013 respectively) than not-calcified plaques, but with the same incidence of intraplaque complications (P=0.156). Accordingly, both calcified and not calcified plaques showed similar mean densities of positivity for CD34, Nestin and WT1. The density of Nestin and WT1 correlated with the occurrence of intra-plaque hemorrhage in all cases, while the density of CD34 correlated only in not-calcified plaques. Conclusions. We confirmed that the Nestin+/WT1- phenotype characterizes the neovessels of instable plaques, regardless the real amount of CD34-positive neoangiogenesis. The calcified plaques show the same incidence of histological complications, albeit they do not influence symptomatology and plaque vulnerability. Female patients show a much higher incidence of not-complicated or calcified plaques, receiving de facto a sort of protection compared to male patients.

La distrofia muscolare di Emery-Dreifuss (EDMD) è una miopatia degenerativa ereditaria caratterizzata da debolezza e atrofia dei muscoli senza coinvolgimento del sistema nervoso. Individui EDMD presentano, inoltre, cardiomiopatia con difetto di conduzione che provoca rischio di morte improvvisa. Diversi studi evidenziano un coinvolgimento di citochine in diverse distrofie muscolari causanti infiammazione cronica, riassorbimento osseo, necrosi cellulare. Abbiamo effettuato una valutazione simultanea della concentrazione di citochine, chemochine, fattori di crescita, presenti nel siero di un gruppo di 25 pazienti EDMD. L’analisi effettuata ha evidenziato un aumento di citochine quali IL-17, TGFβ2, INF-γ e del TGFβ1. Inoltre, una riduzione del fattore di crescita VEGF e della chemochina RANTES è stata rilevata nel siero dei pazienti EDMD rispetto ai pazienti controllo. Ulteriori analisi effettuate tramite saggio ELISA hanno evidenziato un aumento dei livelli di TGFβ2 e IL-6 nel terreno di coltura di fibroblasti EDMD2. Per testare l’effetto nei muscoli, di citochine alterate, abbiamo utilizzato terreno condizionante di fibroblasti EDMD per differenziare mioblasti murini C2C12. Una riduzione del grado di differenziamento è stata osservata nei mioblasti condizionati con terreno EDMD. Trattando queste cellule con anticorpi neutralizzanti contro TGFβ2 e IL-6 si è avuto un miglioramento del grado di differenziamento. In C2C12 che esprimevano la mutazione H222P del gene Lmna,non sono state osservate alterazioni di citochine e benefici di anticorpi neutralizzanti. I dati mostrano un effetto patogenetico delle citochine alterate come osservato in fibroblasti e siero di pazienti, suggerendo un effetto sul tessuto fibrotico di muscoli EDMD. Un effetto intrinseco alla mutazione della lamina A è stato rilevato sul espressione di caveolina 3 in mioblasti differenziati EDMD. I risultati si aggiungono a dati forniti sulla patogenesi dell' EDMD confermando che fattori intrinseci ed estrinseci contribuiscono alla malattia. Utilizzo di anticorpi neutralizzanti specifici contro fattori estrinseci potrebbe rappresentare un approccio terapeutico come mostrato in questo studio. / Emery-Dreifuss muscular dystrophy is a degenerative myopathy characterized by weakness and atrophy of muscle without involvement of the nervous system. Moreover, EDMD patients present cardiomypathy with conduction defects causing risk of sudden death. Different studies highlighted cytokine involvement in muscular dystrophy, causing chronic inflammation, bone resorption or cellular necrosis. We carried out a simultaneous assessment of the concentration of 24 secreted molecules using a wide screening approach. We tested serum concentrations of cytokines, chemokines and growth factors in 25 EDMD patients. Analysis showed an increase of IL-17, TGFβ2, INFγ, and TGFβ1. Furthermore, decrease of VEGF and of the chemokine Rantes was observed in EDMD patients. Further analysis displayed an increase of TGFβ2 and IL-6 levels in culture medium of EDMD skin fibroblasts. To test the effect of altered cytokine levels in muscle, we used conditioned medium from EDMD fibroblasts to culture differentiating mouse myoblasts. Reduced rate of myoblast differentiation was observed in the presence of EDMD conditioning media. Moreover, neutralizing antibodies against TGFβ2 and IL-6 rescued myogenic differentiation. In C2C12 mouse myoblasts expressing the H222P Lmna mutation, we did not observe altered cytokine levels or beneficial effects of neutralizing antibodies. These results show a pathogenetic effect of the altered secretory phenotype here observed in fibroblasts and serum from EDMD patients, hinting at a major role of fibrotic tissue in muscle misfunctioning in EDMD. Furthermore, we observed an intrinsic effect of lamin A mutations on the expression of caveolin 3 in differentiating EDMD myoblasts. Our data add to the existing knowledge on the complex EDMD pathogenesis and confirm that cell intrinsic and extrinsic factors contribute to disease. Neutralization of extrinsic factors by specific antibodies, as shown in this study, may represent a possible therapeutic perspective.

La studio dell’Anatomia umana presenta una varietà di sfaccettature, che sono alla base della reale comprensione del corpo umano; ovvero la vera anatomia non è quella rappresentata nei testi ma quella che appare durante la dissezione o nelle più sofisticate analisi di immagine. Lo scopo di questa tesi è stato quello di rivisitare alcune situazioni vascolari che possono andare incontro a variazioni e cercare di comprendere, anche con l’aiuto della bibliografia, se tali variazioni possono essere causa o epifenomeni di patologie a carico delle arterie affette dalle variazioni stesse o di territori da esse dipendenti per l’afflusso sanguigno. E’ stata condotta una analisi su preparati cadaverici in particolare in tre distretti: a) addome e tripode celiaco/mesenterica superiore; b) circolo cerebrale; d) orco aortico. / The study Human Anatomy presents a variety of facets, which are the basis of the real understanding of the human body; i.e. the real anatomy that is not represented in the texts but one that appears during dissection or in the more sophisticated image analysis. The purpose of this thesis was to revisit some situations that are likely to experience vascular changes and try to understand, also with the help of the bibliography, if these changes can be cause or epiphenomena of pathologies of the arteries affected by the variations or of territories that they supply. It has been carried out an analysis on cadavers and namely in three districts: a) abdomen and celiac trunk/ superior mesenteric artery; b) cerebral circulation; d) aortic arch.

Through the years, several studies reported the involvement of nuclear lipid signalling as highly connected with cell cycle progression. Indeed, nuclear Phosphatidylinositol-4,5-Biphosphate (PIP2) hydrolisis mediated by Phospholipases C (PLC), which leads to production of the second messengers Diacylglycerol (DAG) and Inositol-1,4,5-Triphosphate (IP3), is a fundamental event for both G1/S and G2/M checkpoints. In particular, we found that nuclear DAG production was mediated by PLCbeta1, enzyme mainly localized in the nucleus of K562 human erythroleukemia cells. This event triggered the activation and nuclear translocation of PKCalpha, which, in turn, resulted able to affect cell cycle via modulation of Cyclin D3 and Cyclin B1, two important enzymes for G1/S transition and G2/M progression respectively.