Changes in tumor stiffness for early prediction of tumor response to sorafenib: a proof-of-concept study with elastosonography in an animal model of Hepatocellular Carcinoma (HCC)

Background and aims: Sorafenib is the reference therapy for advanced Hepatocellular Carcinoma (HCC). No method exists to predict in the very early period subsequent individual response. Starting from the clinical experience in humans that subcutaneous metastases may rapidly change consistency under sorafenib and that elastosonography a new ultrasound based technique allows assessment of tissue stiffness, we investigated the role of elastonography in the very early prediction of tumor response to sorafenib in a HCC animal model. Methods: HCC (Huh7 cells) subcutaneous xenografting in mice was utilized. Mice were randomized to vehicle or treatment with sorafenib when tumor size was 5-10 mm. Elastosonography (Mylab 70XVG, Esaote, Genova, Italy) of the whole tumor mass on a sagittal plane with a 10 MHz linear transducer was performed at different time points from treatment start (day 0, +2, +4, +7 and +14) until mice were sacrified (day +14), with the operator blind to treatment. In order to overcome variability in absolute elasticity measurement when assessing changes over time, values were expressed in arbitrary units as relative stiffness of the tumor tissue in comparison to the stiffness of a standard reference stand-off pad lying on the skin over the tumor. Results: Sor-treated mice showed a smaller tumor size increase at day +14 in comparison to vehicle-treated (tumor volume increase +192.76% vs +747.56%, p=0.06). Among Sor-treated tumors, 6 mice showed a better response to treatment than the other 4 (increase in volume +177% vs +553%, p=0.011). At day +2, median tumor elasticity increased in Sor-treated group (+6.69%, range –30.17-+58.51%), while decreased in the vehicle group (-3.19%, range –53.32-+37.94%) leading to a significant difference in absolute values (p=0.034). From this time point onward, elasticity decreased in both groups, with similar speed over time, not being statistically different anymore. In Sor-treated mice all 6 best responders at day 14 showed an increase in elasticity at day +2 (ranging from +3.30% to +58.51%) in comparison to baseline, whereas 3 of the 4 poorer responders showed a decrease. Interestingly, these 3 tumours showed elasticity values higher than responder tumours at day 0. Conclusions: Elastosonography appears a promising non-invasive new technique for the early prediction of HCC tumor response to sorafenib. Indeed, we proved that responder tumours are characterized by an early increase in elasticity. The possibility to distinguish a priori between responders and non responders based on the higher elasticity of the latter needs to be validated in ad-hoc experiments as well as a confirmation of our results in humans is warranted.

Characterization of circulating steroids and endocannabinoids levels in obese subjects by liquid chromatography tandem mass spectrometry

Nowadays obesity can be defined as a global epidemic. The precise identification of circulating biomarkers involved in this pathology could be essential to early diagnose potential co-morbidities and to better address the development of future therapeutic strategies. Published evidences show that circulating steroid hormones and endocannabinoids might have a role in the physiopathology of obesity; however, a precise and reliable quantification of these molecules is still lacking. In the first part of the present thesis, we developed a sensitive, specific and accurate quantification method for nine steroid hormones using a liquid chromatography tandem mass spectrometry (LC-MS/MS) system. This method has been used first for a comparative study with immunoassays, currently used in the clinical practice to quantify these molecules and then to redefine circulating reference intervals in healthy subjects. Furthermore, we measured circulating steroid hormones in three groups of subjects: normo-weight, over-weight and obese, defining different steroid hormones profiles depending on the obesity state. The role of circulating endocannabinoids in humans is still unclear, however there are several evidences concerning their involvement in obesity. In the second part of the thesis, we determined changes of circulating endocannabinoids in obese patients after a weight loss induced by bariatric surgery, currently the most effective long-term treatment for obesity, using LC/MS-MS. We measured basal and dynamic endocannabinoids plasma levels in 12 patients with severe obesity before, one month after and six months after the Roux-en-Y gastric bypass intervention, currently one of the most performed types of bariatric surgery. All together the findings illustrated in this thesis project will help better define the role of steroid hormones and endocannabinoids in the framework of obesity in humans and the role that each type of molecule might have in its pathophysiology.

Parametri emato-biochimici e contaminanti ambientali in Tartarughe e Chirotteri

La biochimica clinica dei rettili e dei cheloni in particolare, non ha ottenuto al momento lo stesso livello di attenzione rivolto ai mammiferi. Con il presente lavoro viene proposta la valutazione dei più importanti parametri ematologici e biochimici in campioni di sangue di Testudo hermanni al fine di facilitare l'interpretazione dei dati di laboratorio e la diagnosi di eventuali patologie. A questo scopo, sono stati calcolati gli intervalli di riferimento di questi parametri in individui clinicamente sani e sono state analizzate le influenze di fattori ambientali e fisiologici. Sono state inoltre determinate le concentrazioni di alcuni importanti elementi chimici sia essenziali che non essenziali. Su campioni di chirottero del genere Tadarida teniotis sono state determinate le concentrazioni di PCB DL, PCB NDL, PCDD/F, PFAS e di elementi chimici essenziali e non, al fine di valutare: il possibile livello di contaminazione in relazione alla specie e all’habitat, la prevalenza e/o il rapporto delle classi di composti nel caso di una possibile contaminazione, la eventuale fonte di contaminazione. A quanto ci risulta, il nostro lavoro rappresenta il primo tentativo di analizzare su vasta scala le concentrazioni di questi inquinanti in una popolazione di pipistrelli residenti in un’area urbana. Tra i vari contaminanti esaminanti in questo studio, il Pb e le diossine possono costituire un serio problema per Tadarida teniotis.

Tetti verdi: Analisi sperimentale e Modellazione numerica

I tetti verdi rappresentano, sempre più frequentemente, una tecnologia idonea alla mitigazione alle problematiche connesse all’ urbanizzazione, tuttavia la conoscenza delle prestazioni dei GR estensivi in clima sub-Mediterraneo è ancora limitata. La presente ricerca è supportata da 15 mesi di analisi sperimentali su due GR situati presso la Scuola di Ingegneria di Bologna. Inizialmente vengono comparate, tra loro e rispetto a una superficie di riferimento (RR), le prestazioni idrologiche ed energetiche dei due GR, caratterizzati da vegetazione a Sedum (SR) e a erbe native perenni (NR). Entrambi riducono i volumi defluiti e le temperature superficiali. Il NR si dimostra migliore del SR sia in campo idrologico che termico, la fisiologia della vegetazione del NR determina l'apertura diurna degli stomi e conseguentemente una maggiore evapotraspirazione (ET). Successivamente si sono studiate la variazioni giornaliere di umidità nel substrato del SR riscontrando che la loro ampiezza è influenzata dalla temperatura, dall’umidità iniziale e dalla fase vegetativa. Queste sono state simulate mediante un modello idrologico basato sull'equazione di bilancio idrico e su due modelli convenzionali per la stima della ET potenziale combinati con una funzione di estrazione dell’ umidità dal suolo. Sono stati proposti dei coefficienti di correzione, ottenuti per calibrazione, per considerare le differenze tra la coltura di riferimento e le colture nei GR durante le fasi di crescita. Infine, con l’ausilio di un modello implementato in SWMM 5.1. 007 utilizzando il modulo Low Impact Development (LID) durante simulazioni in continuo (12 mesi) si sono valutate le prestazioni in termini di ritenzione dei plot SR e RR. Il modello, calibrato e validato, mostra di essere in grado di riprodurre in modo soddisfacente i volumi defluiti dai due plot. Il modello, a seguito di una dettagliata calibrazione, potrebbe supportare Ingegneri e Amministrazioni nella valutazioni dei vantaggi derivanti dall'utilizzo dei GR.

Kinetics of photogenerated carriers in nanostructured semiconductor heterojunctions for solar water splitting

This thesis aims to investigate the fundamental processes governing the performance of different types of photoelectrodes used in photoelectrochemical (PEC) applications, such as unbiased water splitting for hydrogen production. Unraveling the transport and recombination phenomena in nanostructured and surface-modified heterojunctions at a semiconductor/electrolyte interface is not trivial. To approach this task, the work presented here first focus on a hydrogen-terminated p-silicon photocathode in acetonitrile, considered as a standard reference for PEC studies. Steady-state and time-resolved excitation at long wavelength provided clear evidence of the formation of an inversion layer and revealed that the most optimal photovoltage and the longest electron-hole pair lifetime occurs when the reduction potential for the species in solution lies within the unfilled conduction band states. Understanding more complex systems is not as straight-forward and a complete characterization that combine time- and frequency-resolved techniques is needed. Intensity modulated photocurrent spectroscopy and transient absorption spectroscopy are used here on WO3/BiVO4 heterojunctions. By selectively probing the two layers of the heterojunction, the occurrence of interfacial recombination was identified. Then, the addition of Co-Fe based overlayers resulted in passivation of surface states and charge storage at the overlayer active sites, providing higher charge separation efficiency and suppression of recombination in time scales that go from picoseconds to seconds. Finally, the charge carrier kinetics of several different Cu(In,Ga)Se2 (CIGS)-based architectures used for water reduction was investigated. The efficiency of a CIGS photocathode is severely limited by charge transfer at the electrode/electrolyte interface compared to the same absorber layer used as a photovoltaic cell. A NiMo binary alloy deposited on the photocathode surface showed a remarkable enhancement in the transfer rate of electrons in solution. An external CIGS photovoltaic module assisting a NiMo dark cathode displayed optimal absorption and charge separation properties and a highly performing interface with the solution.

Musical tension in harmonic intervals: behavioral and neural correlates

Musical tension is what drives our emotional experience in music listening. However, the specific role of the musical elements involved in tension-resolution perception remains largely unclear. This dissertation aims to advance the understanding of tension perception dynamics related to sensory consonance-dissonance. The first experiment aimed to design and validate a new crossmodal proprioceptive device for tension rating that overcomes some of the limitations of known tools. As a result, a psychophysical equation for the matching of physical force and psychological force was presented. The same tool was subsequently used in the second and third experiments to collect ratings of perceived tension and movement in harmonic musical intervals and standard noises. Besides, a visual analog scale (VAS) was used to allow a comparison of these two methods. The results confirmed the close relationship between sensory dissonance and perceived tension. Moreover, stimuli in the higher pitch register were perceived as more tense, confirming the primary role of pitch as a mediator of tension. The comparison between ratings obtained with the proprioceptive device and the VAS highlighted the tendency to give higher tension ratings using the VAS compared to the proprioceptive device. In the last experiment, brain electrical activity was recorded during the presentation of short tension-resolution patterns created using the most tense (perfect unison, fourth, and fifth) and the least tense harmonic intervals (augmented fourth, minor second, and inverted major seventh) to understand how consonance-dissonance can convey meaningful information on perceived tension-resolution. Results showed overall larger effects during the ‘resolution’ condition compare to the ‘tension induction’ condition, indicating that the resolution of harmonic instability towards a state of stability may be more salient than its opposite. A late positive component (LPC) was elicited, possibly reflecting deeper processing of tension-related meaning within a minimal harmonic context.