Il trapianto di pancreas nel trattamento del diabete mellito tipo 1

Il trapianto di pancreas rappresenta un’alternativa terapeutica di interesse nel trattamento dei pazienti con diabete mellito tipo 1. Tuttavia la considerazione del delicato rapporto rischi/benefici rende necessarie ulteriori informazioni circa i risultati a lungo termine associati a tale procedura. Nel nostro centro il programma di TP, iniziato nel 1996 con l’esecuzione del primo trapianto combinato rene-pancreas (SPK), dal 2000 ha introdotto il trapianto di pancreas isolato (PTA) e dal 2002 il trapianto di pancreas dopo rene (PAK). Complessivamente sono stati eseguiti 355 trapianti in 331 pazienti (età: 39.0±8 anni; rapporto maschi/femmine, 192/163; IMC 23.2±2.8 kg/m2; durata del diabete, 24±8 anni) selezionati secondo le indicazioni fornite dalle associazioni mediche nazionali ed internazionali. I trapianti eseguiti erano così suddivisi: 231 SPK, 93 PTA e 31 PAK. Nel gruppo SPK, 78 (34.5%) sono stati eseguiti in pazienti affetti da diabete mellito tipo 1 ed insufficienza renale terminale in fase pre-dialitica. Nel 50% dei casi il trapianto è stato eseguito con derivazione enterico-portale, nel 39% con derivazione enterico-sistemica e nel restante 11% con drenaggio di tipo enterico-vescicale. La terapia antirigetto è consistita in una fase di induzione con basiliximab (63%) o siero antilinfocitario (37%), seguita da terapia cronica con tacrolimo (81%) o ciclosporina (19%), micofenolato alla dose terapeutica massima tollerata, e basse dosi di deltacortene (5 mg/die). Complessivamente la sopravvivenza attuariale del paziente a 1, 3, 5, e 10 anni è risultata rispettivamente del 96, 95, 94 e 92%. Agli stessi tempi la sopravvivenza del pancreas, intesa in termini di insulino-indipendenza, è risultata pari all’87, 79, 76 e 75% nei PTA e all’88, 85, 83 e 81% nell’SPK. Di tutti i trapianti eseguiti, 107 SPK e 34 PTA, sono stati eseguiti entro il 31 dicembre 2003. Di questi pazienti sono stati analizzati i risultati metabolici prima del trapianto e ad 1,5 e 10 anni post trapianto, in termini di glicemia, emoglobina glicosilata, peptide C ed insulinemia, profilo lipidico. La valutazione della funzione renale è avvenuta mediante misurazioni seriate dei valori di creatinina plasmatica e la valutazione del filtrato glomerulare calcolata secondo la formula MDRD. A 10 anni dal trapianto, la sopravvivenza attuale dei pazienti è risultata pari al 97% sia nei pazienti sottoposti a SPK che a PTA, mentre quella del pancreas pari al 73% nei trapianti combinati e al 63% nei trapianti isolati. (Nei soggetti con organo funzionante, il ripristino della secrezione insulinica endogena (peptide C a 10 anni: 2.81±1.47 ng/ml, p<0.001 vs valori pre-trapianto: 0.08±0.11 ng/ml) ha consentito il conseguimento e il mantenimento di normali valori di glicemia a digiuno (91±10 a 10 anni vs 191±9 mg /dl pre trapianto negli SPK e agli stessi tempi 96±19 vs 230±108mg/dl nei PTA) e di HbA1c (5.8±0.7 vs 8.7±2.0 negli SPK e 5.7±0.5 vs 8.3±1.8% nei PTA, a 10 anni vs pre-Trapianto) (tutti p<0.001 rispetto ai valori pre-trapianto). Il colesterolo totale (212±54 vs 178±33 negli SPK, 157±40 vs 193±31 mg/dl, nei PTA, p<0.001) e quello LDL (132±44 vs 100±30 negli SPK e 95±36 vs 128±36 mg/dl nei PTA, p<0.05) sono risultati anch’essi ridotti significativamente, senza modifiche sostanziali nella terapia con statine. Nei pazienti sottoposti a SPK si è osservata una riduzione significativa anche dei valori di trigliceridi (109.6±47.6 a 10 anni vs 175.1±82.8 pre-trapianto, p<0.001). La funzionalità renale, stimata mediante MDRD, ha mostrato una riduzione del filtrato glomerulare (GFR) pari a 1.8±2 ml/min/anno nei PTA e del 1.34±1.8 ml/min/annuo. La diminuzione del filtrato annua è risultata significativa solo nei pazienti sottoposti a PTA con filtrato superiore a 90 ml/min prima del trapianto nei primi due anni per poi stabilizzarsi. I risultati di questo studio monocentrico mostrano una buona sopravvivenza a lungo termine dei pazienti e degli organi trapiantati, tali da confortare circa l’efficacia e la ragionevole sicurezza di queste procedure

Insulin secretory granules labelled with phogrin-fluorescent proteins show alterations in size, mobility and responsiveness to glucose stimulation in living β-cells

The intracellular life of insulin secretory granules (ISGs) from biogenesis to secretion depends on their structural (e.g. size) and dynamic (e.g. diffusivity, mode of motion) properties. Thus, it would be useful to have rapid and robust measurements of such parameters in living β-cells. To provide such measurements, we have developed a fast spatiotemporal fluctuation spectroscopy. We calculate an imaging-derived Mean Squared Displacement (iMSD), which simultaneously provides the size, average diffusivity, and anomalous coefficient of ISGs, without the need to extract individual trajectories. Clustering of structural and dynamic quantities in a multidimensional parametric space defines the ISGs’ properties for different conditions. First, we create a reference using INS-1E cells expressing proinsulin fused to a fluorescent protein (FP) under basal culture conditions and validate our analysis by testing well-established stimuli, such as glucose intake, cytoskeleton disruption, or cholesterol overload. After, we investigate the effect of FP-tagged ISG protein markers on the structural and dynamic properties of the granule. While iMSD analysis produces similar results for most of the lumenal markers, the transmembrane marker phogrin-FP shows a clearly altered result. Phogrin overexpression induces a substantial granule enlargement and higher mobility, together with a partial de-polymerization of the actin cytoskeleton, and reduced cell responsiveness to glucose stimulation. Our data suggest a more careful interpretation of many previous ISG-based reports in living β-cells. The presented data pave the way to high-throughput cell-based screening of ISG structure and dynamics under various physiological and pathological conditions

Co-localization of acinar markers and insulin in pancreatic cells of subjects with type 2 diabetes

To search for clues suggesting that beta cells may generate by transdifferentiation in humans, we assessed the presence of cells double positive for exocrine (amylase, carboxypeptidase A) and endocrine (insulin) markers in the pancreas of non-diabetic individuals (ND) and patients with type 2 diabetes (T2D). Samples from twelve ND and twelve matched T2D multiorgan donors were studied by electron microscopy, including amylase and insulin immunogold labeling; carboxypeptidase A immunofluorescence light microscopy assessment was also performed. In the pancreas from four T2D donors, cells containing both zymogen-like and insulin-like granules were observed, scattered in the exocrine compartment. Nature of granules was confirmed by immunogold labeling for amylase and insulin. Double positive cells ranged from 0.82 to 1.74 per mm2, corresponding to 0.26±0.045% of the counted exocrine cells. Intriguingly, cells of the innate immune systems (mast cells and/or macrophages) were adjacent to 33.3±13.6% of these hybrid cells. No cells showing co-localization of amylase and insulin were found in ND samples by electron microscopy. Similarly, cells containing both carboxypeptidase A and insulin were more frequently observed in the diabetic pancreata. These results demonstrate more abundant presence of cells containing both acinar markers and insulin in the pancreas of T2D subjects, which suggests possible conversion from one cellular type to the other and specific association with the diseased condition

APOLLO 11 Project, Consortium in Advanced Lung Cancer Patients Treated With Innovative Therapies: Integration of Real-World Data and Translational Research

Introduction: Despite several therapeutic efforts, lung cancer remains a highly lethal disease. Novel therapeutic approaches encompass immune-checkpoint inhibitors, targeted therapeutics and antibody-drug conjugates, with different results. Several studies have been aimed at identifying biomarkers able to predict benefit from these therapies and create a prediction model of response, despite this there is a lack of information to help clinicians in the choice of therapy for lung cancer patients with advanced disease. This is primarily due to the complexity of lung cancer biology, where a single or few biomarkers are not sufficient to provide enough predictive capability to explain biologic differences; other reasons include the paucity of data collected by single studies performed in heterogeneous unmatched cohorts and the methodology of analysis. In fact, classical statistical methods are unable to analyze and integrate the magnitude of information from multiple biological and clinical sources (eg, genomics, transcriptomics, and radiomics). Methods and objectives: APOLLO11 is an Italian multicentre, observational study involving patients with a diagnosis of advanced lung cancer (NSCLC and SCLC) treated with innovative therapies. Retrospective and prospective collection of multiomic data, such as tissue- (eg, for genomic, transcriptomic analysis) and blood-based biologic material (eg, ctDNA, PBMC), in addition to clinical and radiological data (eg, for radiomic analysis) will be collected. The overall aim of the project is to build a consortium integrating different datasets and a virtual biobank from participating Italian lung cancer centers. To face with the large amount of data provided, AI and ML techniques will be applied will be applied to manage this large dataset in an effort to build an R-Model, integrating retrospective and prospective population-based data. The ultimate goal is to create a tool able to help physicians and patients to make treatment decisions. Conclusion: APOLLO11 aims to propose a breakthrough approach in lung cancer research, replacing the old, monocentric viewpoint towards a multicomprehensive, multiomic, multicenter model. Multicenter cancer datasets incorporating common virtual biobank and new methodologic approaches including artificial intelligence, machine learning up to deep learning is the road to the future in oncology launched by this project

Aproximación al mural en hormigón de Javier Clavo de la antigua facultad de Ingenieros Agrónomos: Diálogo entre obra mural y arquitectura

El objeto de este estudio es analizar la figura del artista madrileño Javier Clavo (1918-1994) centrándose en la obra mural en hormigón armado de la antigua facultad de Ingenieros Agrónomos de Blasco Ibáñez, contextualizada en el marco de la arquitectura moderna del arquitecto Fernando Moreno Barberá.Occhipinti, M. (2011). Aproximación al mural en hormigón de Javier Clavo de la antigua facultad de Ingenieros Agrónomos: Diálogo entre obra mural y arquitectura. http://hdl.handle.net/10251/15305Archivo delegad