L'ipossia è una condizione patologica determinata da un ridotto apporto di ossigeno a livello tissutale. A partire dagli anni '60, Tomlinson e Gray hanno dimostrato la presenza di regioni di scarsa ossigenazione nel cancro del polmone e hanno osservato che questa caratteristica del microambiente tumorale è associata ad una maggiore resistenza alla radioterapia. La determinazione dell'ipossia nei tumori è pertanto della massima rilevanza clinica, in quanto l'aggressività del tumore, la deriva metastatica, il mancato controllo della neoplasia, l'aumento del rischio di recidiva e, in definitiva l'esito sfavorevole sono associati all'ipossia. Negli ultimi decenni, c'è stato un crescente interesse nello sviluppo di metodi per la valutazione dell'ossigenazione tumorale. Questi metodi possono essere invasivi, come il sensore polarografico di O2, o non invasivi, basati principalmente sulle tecniche di imaging. Le modalità di imaging sono senza dubbio le più interessanti perché garantiscono una visualizzazione onnicomprensiva del tessuto patologico e sono in grado di identificare il fenomeno anche in luoghi inaccessibili alle procedure invasive. Tra le modalità di imaging per la valutazione dell'ipossia, la tomografia ad emissione di positroni (PET) è una delle più studiate, in quanto offre: (a) ampio assortimento di radiofarmaci; (B) buona risoluzione intrinseca; (C) rappresentazione tridimensionale (3D); (D) (semi)quantificazione; (E) maggiore facilità per il paziente, e (f) riproducibilità dei dati. L'oggetto della tesi attuale è quello di studiare il ruolo dell'imaging molecolare alla PET nell'ipossia tumorale. Il testo è diviso in quattro sessioni distinte incentrate nel fornire in primis una panoramica sui radiofarmaci principali (Sessione-1), poi nella valutazione della correlazione tra l'espressione tissutale d'ipossia e l'imaging alla 18F-FDG PET/TC (Sessione-2). Le altre due sessioni analizzano l'impatto prognostico del tracciante per l'ipossia (64Cu-ATSM) nei tumori solidi (Sessione-3), seguito da una sofisticata analisi frattale di confronto fra le acquisizioni precoci e tardive alla 64Cu ATSM PET/TC nei tumori solidi (Sessione-4). / Hypoxia is a pathological condition caused by a reduced oxygen supply at the tissue level. Since the 60’s, Tomlinson and Gray have demonstrated the presence of regions of poor oxygenation in lung cancer, and noted that this characteristic of tumor microenvironment is associated to increased resistance to radiotherapy. The detection of hypoxia in tumors is therefore of utmost clinical relevance, because tumor aggressiveness, metastatic drift, failure to disease control, increased risk of recurrence and ultimately poor outcome are associated with hypoxia. In recent decades, there has been an increasing interest in developing methods for the assessment of tumor oxygenation. These methods can be invasive, such as the polarographic O2-sensor, or non-invasive, mainly based on imaging techniques. Imaging modalities are undoubtedly the most appealing techniques for this purpose, because they guarantee an all-encompassing visualization of the pathological tissue and can identify the phenomenon even at sites inaccessible to invasive procedures. Among the image-based modalities for hypoxia assessment, positron emission tomography (PET) is one of the most extensively investigated, because it offers: (a) broad assortment of radiopharmaceuticals; (b) good intrinsic resolution; (c) three-dimensional (3D) tumor representation; (d) (semi)quantification of the hypoxic burden; (e) patient friendliness, and (f) reproducibility. The object of the current thesis is to investigate the role of molecular imaging with PET in cancer hypoxia. The text is divided into four different sessions focused on giving at first an insight on principal radiopharmaceuticals applied for hypoxia imaging (Session-1), then concentrating on the correlation of tissue expression of hypoxia and imaging findings on 18F-FDG PET/CT (Session-2). The next two sessions will analyze the prognostic impact of the hypoxia-specific tracer (64Cu-ATSM) in solid tumors (Session-3), followed by a sophisticated ad hoc computer-aided fractal geometry based analysis of DICOM images for early and late acquisitions on 64Cu-ATSM PET/CT in solid tumors (Session-4).

Background: An increasing number of studies have established that circulating tumor cells (CTCs) are a heterogeneous population in which cells have different degrees of metastatic potential mainly due to epithelial-mesenchymal transition (EMT). This study aimed to assess the feasibility of circulating esophageal cancer cells identification, characterization and to evaluate their prognostic value in esophageal cancer Methods: In order to closely mimic CTC heterogeneity, MC10A and HMEL cell lines differently forced in EMT are used. Single cell analysis was conducted by DEPArray. We assigned a specific phenotypic tag to each potential CTC population: Epithelial-tag, Mesenchymal/stem-tag. We evaluated the basal cell phenotype and after EMT induction. Subsequently, the Grab all assay was performed on peripheral blood samples from patients with esophageal cancer. This feasibility study enrolled 11 patients (4 M1, 7 M0). Analyses were conducted on 3 peripheral blood samples (15/20 ml) per patients. Blood samples from non-metastatic patients were taken before and after primary neo-adjuvant therapy and after secondary treatment (surgery). Samples from metastatic patients were taken before and after first line therapy and after second line therapy. CTCs were identified and sorted singly by DEPArray system. Results: The assay was able to detect the phenotypic changes in cell lines mimicking CTC heterogeneity. Before therapy, CTCs were found in 3/4 metastatic patients. Of the 7 non-metastatic patients, 3 were positive for CTCs before therapy. Examining CTC status at different clinical time points, it was possible to suggest a correlation between the presence of CTCs and disease progression. Conclusions: Data showed that the assay is feasible, capable to analyze the phenotypic tags by DEPArray using a multiple staining without aspecific signals. Experiments carried out from both metastatic/non metastatic cancer patients showed the ability of the Grab-allassay to identify subpopulations of CTCs with different epithelial/stem/mesenchymal or hybrid phenotypes potentially related to disease progression.

Nowadays video-assisted lobectomy could be considered an appropriate treatment option for early stage lung cancer patients, as obtaining similar or even better early outcomes when compared to open standard lobectomy. This study was aimed at verifying the long-term impact (three months after surgery) of VATS-lobectomy technique on physical and mental status of patients undergoing lung resection for lung cancer and to compare the physical variation to the one registered after open approach. We enrolled patients treated by VATS-lobectomy at our institution since June 2012 to June 2015, comparing the preoperative physical evaluation (in terms of FEV1, DLCO and VO2max) and perceived quality of life (measured using the EORTC questionnaire) to the postoperative values. Moreover we performed a propensity score matching analysis to verify if the physical variation registered after VATS-lobectomy differed from that after open resection. We found a three months postoperative reduction of the physical indicators in comparison to the preoperative values (FEV1-variation: -8.3%, DLCO-variation: -12.4%, VO2max-variation: -6.6%). A similar negative trend was found for the psychological indicators, but the only parameter with a significative worsening was the perceived physical functioning (variation: -6.2%). The propensity score procedure yielded 83 well-matched open and VATS-lobectomy patients. The reductions in FEV1, DLCO and VO2max were similar in both groups (open patients’ reduction: FEV1 -10%, DLCO -11.9%, VO2max -5.5%; VATS patients’ reduction: FEV1 -7.2%, DLCO -10.6%, VO2max -6.9%), with a Cohen effect size < 0.2 for all the comparisons. In conclusion, VATS-lobectomy influences a reduction of the preoperative functional status three months after the operation, without affecting the postoperative perceived quality of life. Moreover VATS-lobectomy doesn’t offer any advantage in terms of FEV1, DLCO and exercise capacity recovery in comparison to the open approach.

Oggetto della tesi e' l'approfondimento su tecniche e metodiche di preservazione del polmone isolato per lo studio ecografico. E' discussa l'appropriatezza sull'uso degli ultrasuoni in corso di chirurgia mini invasiva polmonare, obiettivo di una ricerca sperimentale. Il razionale dello studio si fonda sull'indicazione all'exeresi chirurgica di noduli polmonari di diametro inferiore al centometro, ovvero di diametro superiore ma localizzati in aree centrali del polmone. Queste lesioni sono sempre piu' frequentente diagnosticate per mezzo di avanzate tecniche di imaging. L'atto chirurgico ha scopo terapeutico quando sia stata posta la diagnosi di neoplasia maligna, diagnostico-terapeutico quando non sia ancora ottenuta la tipizzazione istologica della lesione. La tecnica toracoscopica offre numerosi vantaggi rispetto alle tecniche chirurgiche tradizionali ma presenta il grave limite di non permettere la palpazione diretta del tessuto polmonare e la localizzazione della formazione tumorale quando essa non sia visibile macroscopicamente. Gli ultrasuoni sono stati utilizzati con successo per indirizzare la localizzazione del nodulo polmonare. Scopo dello studio sperimentale e' quello di confrontare tecniche diverse di preservazione del polmone isolato in un modello animale, comparando catatteristiche e prestazioni di sonde ecografiche differenti per tipologia. Del tutto recentemente, in ambito trapiantologico, sono state proposte tecniche di preservazione organica utili ai fini di uno studio anatomico sperimentale particolareggiato (EVLP) e moderna e' da considerarsi la concezione di mezzi tecnici specifici per la localizzazione di bersagli intrapolmonari. La tecnica clinica applicata allo studio del particolare ecografico, nel modello animale, ha reso comprensibile e meglio definito il ruolo delle sonde ecografiche nell'individuazione di forme tumorali suscettibili di exeresi definitiva. / The primary focus of this dissertation is research about techniques and methods of isolated lung tissue preservation for sonographic investigation. Doctoral thedis describes the use of ultrasound probes in an experimental model and its clinical application during minimally invasive pulmonary surgery. The rationale of the study is based on the indication to surgical procedure and when is clinically in the treatment of solitary pulmonary nodules (<1 cm or greater but more central in visible appearance). In the era of CT screening programs, these lesions are more frequently diagnosed with advanced imaging techniques. Surgical procedure is therapeutic when a diagnosis of malignancy is obtained, but also diagnostic if we must correct the technique to the histologic pattern or ascertain pathological growth of the lesion. Thoracoscopic techniques offers several advantages over traditional "open" procedures but shows clinical limits. These mininimally invasive approaches do not allow to direct handling of tumor when is not macroscopically visible. Ultrasound have been successfully used to guide localization of pulmonary nodules. The experimental study compare different techniques of lung tissue preservation in a porcine model, the proceed to analysis of diagnostic performances related to a panel of echographic probes. More recently, a number of different techniques, adopted in lung transplantation programs, has been considered as useful for experimental anatomical study (EVLP) and several instruments of scientific research has been comceived for intrapulmomary localization of neoplastic lesions. These clinical techniques were originally applied in an animal porcine model, studied for scientific purpose, and described in a personal review of cases. Results of surgical interesr were related to scientific Literature in order to identify the possible role of echographic probes on clinical context.

Il tumore del polmone e una delle neoplasie più diagnosticate dal 1985 e rimane ancora oggi la causa più frequente di morte cancro-correlata nel mondo. Una resezione polmonare anatomica completa continua ad essere il cardine della terapia per il tumore non a piccole cellule. Perdite aeree prolungate (PAL) sono la più comune complicanza dopo una chirurgia polmonare e sono state riportate con un’incidenza compresa tra il 3-26%, simile sia nelle resezioni polmonari per via toracotomica sia in quelle per via toracoscopica. Fattori di rischio descritti sono scissure interlobari incomplete, patologie polmonari sottostanti (come enfisema, fibrosi, tubercolosi o neoplasie), aderenze pleuriche, pazienti anziani (>75 anni) e bassa capacita di diffusione. Lo sviluppo di strumentazione all’avanguardia e di nuove tecniche chirurgiche ha contribuito a ridurre l’incidenza di queste complicanze. Considerando l’alto impatto clinico e socio-economico di queste problematiche, e stata inoltre sviluppata una varietà di complementari naturali e materiali sintetici molti utili nella gestione delle perdite aeree. / Lung cancer is one of the most diagnosed cancer in 1985 and still remains the most common cause of cancer-related death in the world. A full anatomical lung resection remains the mainstay of therapy for non-small cell cancer. Prolonged air leak (PAL) are the most common complication following a lung surgery and were reported with an incidence of between 3-26%, similar in both pulmonary resection by thoracotomy and in those thoracoscopic. Risk factors described are incomplete interlobar fissures, lung diseases underlying (such as emphysema, fibrosis, tuberculosis or malignancy), pleural adhesions, elderly patients (> 75 years) and low capacity to spread. The development of modern equipment and new surgical techniques has helped to reduce the incidence of these complications. Considering the high clinical impact and socio-economic problems of these, and was also developed a variety of complementary natural and synthetic materials useful in the management of many air leak.

No description available.

No description available.

Gastroesophageal junction (GEJ) adenocarcinoma are uncommon before age of 40 years. While certain clinical, pathological and molecular features of GEJ adenocarcinoma in older patients have been extensively studied, these characteristics in the younger population remain to be determined. In the recent literature, a high sensitivity and specificity for the detection of dysplasia and esophageal adenocarcinoma was demonstrated by using multicolor fluorescence in situ hybridization (FISH) DNA probe set specific for the locus specific regions 9p21 (p16), 20q13.2 and Y chromosome. We evaluated 663 patients with GEJ adenocarcinoma and further divided them into 2 age-groups of <or= 40 and >or= 50 years, rispectively. FISH with selected DNA probe for Y chromosome, locus 9p21 (p16), and locus 20q13.2 was investigated with formalin fixed and parassin embedded tissue from surgical resections of 17 younger and 11 older patients. Signals were counted in > 100 cells with each given histopathological category. The chromosomal aberrations were then compared in the 2 age-groups with the focus on uninvolved squamous and columnar epithelium, intestinal metaplasia (Barrett's mucosa), glandular dysplasia, and adenocarcinoma. Comparisons were performed by the X2 test, Fisher's exact test, Student's t-test and Mann-Whitney U-test as appropriate. Survival was estimated by the Kaplan-Meier method with univariate analysis by the log-rank. Significance was taken at the 5% level. There was no difference in the surgical technique applied in both age groups and most patients underwent Ivor Lewis esophagectomy. Among clinical variables there was a higher incidence of smocking history in older patient group. We identified a progressive loss of Y chromosome from benign squamos epithelium to Barrett's mucosa and glandular dysplasia, and, ultimately, to a near complete loss in adenocarcinoma in both age groups. The young group revealed significantly more losses of 9p21 in both benign and neoplastic cells when compared to the older patients group. In addition, we demonstrated an increase in the percentage of cells showing gain of locus 20q13.2 with progression from benign epithelium through dysplasia to adenocarcinoma with almost the same trend in both the young and the older patients. When compared with the older age-group, younger patients with GEJ adenocarcinoma possess similar known demographics, environmental factors, clinical, and pathologic characteristics. The most commonly detected genetic aberrations of progressive Y chromosomal loss, 9p21 locus loss, and 20q13 gains were similar in the younger and older patients. However the rate of loss of 9p21 is significantly higher in young patients, in both the benign and the neoplastic cells. The loss of 9p21, and possibly, the subsequent inactivation of p16 gene may be one of the molecular mechanisms responsible for the accelerated neoplastic process in young patients.

No description available.

No description available.