Detection of inappropriate cell proliferation in breast epithelium leading to breast cancer

Breast cancer is predominantly caused by unrestrained cell proliferation. Proliferation is a complex process mediated by a network of signals that converge to a point called the ‘initiation of genome replication’ after which either proliferation or cell death could take place. The minichromosome maintenance (MCM) proteins are located at this point and play a pivotal role in regulating DNA replication. The detection of an aberrant level of such proteins can be of use in early breast cancer diagnosis. The main aim of this thesis was to propose a new system to detect inappropriate cell proliferation in breast epithelium. An in vitro model using cancer cell lines was developed to lay the foundation for subsequent studies employing human breast specimens. The application of the in vitro findings in breast excisions allowed assessing of the specificity and sensitivity of the biomarkers to ascertain slowly proliferating neoplastic cells. The most striking finding of this study was the abnormal presence of the MCM proteins in tumour compared to normal tissues with a typical pattern of expression unique for the histological classification of the lesion. The potential of MCM proteins as indicators of cell proliferation defects was further investigated with association studies with Ki-67, Bcl-2 and ER. MCM consistently identified a higher proportion of proliferating cells compared to Ki-67 suggesting that they are interesting markers of the Gi/S-phase. In fact, the MCM proteins start to co-localise in early Gi whereas Ki-67 is almost absent in this phase. Importantly, MCM proteins could recognise not only the proliferating compartment of the tumour but also those cells with replication potential. Based on these findings, the novel MCM biomarkers can be helpful in identifying both malignant and potentially malignant breast tissues. This feature can be useful in predicting patients at risk of tumour progression.Ph

Evaluation of energy metabolism and calcium homeostasis in cells affected by Shwachman-Diamond syndrome

Isomorphic mutation of the SBDS gene causes Shwachman-Diamond syndrome (SDS). SDS is a rare genetic bone marrow failure and cancer predisposition syndrome. SDS cells have ribosome biogenesis and their protein synthesis altered, which are two high-energy consuming cellular processes. The reported changes in reactive oxygen species production, endoplasmic reticulum stress response and reduced mitochondrial functionality suggest an energy production defect in SDS cells. In our work, we have demonstrated that SDS cells display a Complex IV activity impairment, which causes an oxidative phosphorylation metabolism defect, with a consequent decrease in ATP production. These data were confirmed by an increased glycolytic rate, which compensated for the energetic stress. Moreover, the signalling pathways involved in glycolysis activation also appeared more activated; i.e. we reported AMP-activated protein kinase hyper-phosphorylation. Notably, we also observed an increase in a mammalian target of rapamycin phosphorylation and high intracellular calcium concentration levels ([Ca2+]i), which probably represent new biochemical equilibrium modulation in SDS cells. Finally, the SDS cell response to leucine (Leu) was investigated, suggesting its possible use as a therapeutic adjuvant to be tested in clinical trials

In Vivo Noninvasive Detection of Brown Adipose Tissue through Intermolecular Zero-Quantum MRI

The recent discovery of active Brown Adipose Tissue (BAT) in adult humans has opened new avenues for obesity research and treatment, as reduced BAT activity seem to be implicated in human energy imbalance, diabetes, and hypertension. However, clinical applications are currently limited by the lack of non-invasive tools for measuring mass and function of this tissue in humans. Here we present a new magnetic resonance imaging method based on the normally invisible intermolecular multiple-quantum coherence 1H MR signal. This method, which doesn’t require special hardware modifications, can be used to overcome partial volume effect, the major limitation of MR-based approaches that are currently being investigated for the detection of BAT in humans. With this method we can exploit the characteristic cellular structure of BAT to selectively image it, even when (as in humans) it is intimately mixed with other tissues. We demonstrate and validate this method in mice using PET scans and histology. We compare this methodology with conventional 1H MR fat fraction methods. Finally, we investigate its feasibility for the detection of BAT in humans

The role of Clinical Engineers in wound management: a study on VAC Therapy

reservedPartendo dall'esperienza di tirocinio svolta presso l'Ospedale universitario di Padova si discute l'utilizzo e il funzionamento della terapia avanzata VAC therapy, il cui scopo è quello di favorire il processo di guarigione delle ferite. Dopo aver studiato nel dettaglio alcune sue applicazioni si procede con un confronto tra dispositivi, al fine di simulare e fornire una panoramica di ciò che accade in un contesto ospedaliero durante un' indagine di mercato quando è richiesto l'acquisto di nuova apparecchiatura

Detection of inappropriate cell proliferation in breast epithelium leading to breast cancer

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Prevenzione, promozione e diritto di interpello

Gli Autori analizzano la novita del diritto di interpello in materia di servizi ispettivi e di vigilanza, intodotta con la riforma del 2004. Ne viene in particolare sottolineata la confrmità rispetto alla legge delega e ad alcuni principi costituzionali da taluno invocati per sostenerne l'illegittimità. Viene inoltre valorizzato tale istituto nell'ottica di prevenzione e promozione che caratterizza l'impianto della riforma

NPAS3 is a trachealess homolog critical for lung development and homeostasis

Trachealess (Trh) is a PAS domain transcription factor regulating Drosophila tracheogenesis. No other Trh homolog has been associated with a respiratory phenotype. Seeking homolog(s) regulating lung development, we screened murine genomic DNA using trh oligonucleotides, identifying only Npas3. Npas3 mRNA peaks in lung from E10.5 to E13.5, verified by sequencing, with immunostaining in airway epithelial cells. Npas3-null mice have reduced lung branching morphogenesis but are viable prenatally. Npas3-null newborns die in respiratory distress, with diminished alveolarization, decreased Shh, Fgf9, Fgf10, and Bmp4 mRNAs, and increased Spry2, consistent with reduced FGF signaling. Exogenous FGF10 rescues branching morphogenesis in Npas3-null lungs. In promoter reporter assays, NPAS3 directly upregulates Shh and represses Spry2. Npas3+/− mice have a milder lung phenotype, surviving postnatally, but develop emphysema and airways hyperreactivity. Therefore, absence of a developmentally expressed transcription factor can alter downstream gene expression and multiple signaling pathways in organogenesis. NPAS3 haploinsufficiency may also lead to emphysema and asthma

Accelerated Thymic Maturation and Autoreactive T Cells in Bronchopulmonary Dysplasia

Rationale: Bronchopulmonary dysplasia (BPD), a chronic lung disease of newborns triggered by oxygen and barotrauma, is characterized by arrested alveolarization. Increased levels of bombesin-like peptides shortly after birth mediate lung injury: anti-bombesin antibody 2A11 protects against BPD in two baboon models. The role of adaptive immunity in BPD has not been explored previously