Genetics and Genomics of Pulmonary Arterial Hypertension

Pulmonary arterial hypertension is a severe and progressive disorder affecting the blood vessels in the lungs. Typically, symptoms first appear at around 30–40 years of age and, without treatment, can lead to fatal heart disease within a few years. Genetic studies over the past decade have identified numerous genes that contribute to disease progression but, for many sufferers, the underlying genetic cause remains elusive. The collection of reviews and original research articles contained within this book provide an overview of recent advancements in understanding the genetic risk factors for pulmonary arterial hypertension. We further examine the emerging interplay between genetic variants and clinical outcomes, providing a framework for new treatments and improved patient care

A combined approach on the impact of EP300 overexpression on drug resistance, invasion and stemness in breast cancer

Triple negative breast cancer, characterized by absence of estrogen and progesterone receptors and human epidermal growth factor receptor 2 (HER2), is very aggressive, and often progresses to metastasis. The absence of druggable targets implies that conventional chemotherapeutic agents do not work. Recent genomic studies show that epigenetics play a complicated role in maintaining healthy gene expression. This thesis attempted to demonstrate a link between E1A binding protein P300 (EP300), epithelial to mesenchymal transition (EMT), drug resistance and cancer stem cell (CSC) traits by using breast cancer cell line models with EP300 overexpression (CAL51 and MDA-MB-231), short hairpin RNA (shRNA) lentiviral knockdown EP300 (T47D and MCF7), and EP300 knockout in colorectal cancer (HCT116). Techniques such as flow cytometry, migration/invasion assays and short/long term drug resistance assays were used to demonstrate these traits. This study also employed reverse transcription quantitative real-time PCR (RTqPCR) and RNA sequencing (RNA-seq) gene expression analysis for evaluating a gene signature and miR expression. The study demonstrated positive regulation of EP300, forkhead box O3a (FOXO3a) and GATA binding protein 3 (GATA3) by miR-25 in minimally transformed mammary epithelial cancer (MTMEC) model. Neither RNA-seq nor RTqPCR demonstrated any correlation between cancer subtype and our gene signature. However, we confirmed that EP300 overexpression resulted in E-cadherin (CDH1) upregulation in CAL51 and MDA-MB-231. EP300 knockdown in T47D resulted in decreased expression of CDH1 and upregulated expression of FOXO3a. EP300 knockout in HCT116 downregulated CDH1. EP300 overexpression in MDA-MB-231 improved drug sensitivity to paclitaxel and doxorubicin. EP300 overexpression reduced aldehyde dehydrogenase + (ALDH) populations in MDA-MB-231 and CAL-51 cell lines, while EP300 modulation had a variable effect on CD44 antigen (CD44)high/CD24 antigen (CD24)low subpopulations in these cell line models. Finally, overexpression of EP300 in MDA-MB-231 demonstrated a decrease in motility and invasion.Open Acces

Imaging studies of peripheral nerve regeneration induced by porous collagen biomaterials

Thesis: Ph. D., Massachusetts Institute of Technology, Department of Mechanical Engineering, 2013.Cataloged from PDF version of thesis.Includes bibliographical references.There is urgent need to develop treatments for inducing regeneration in injured organs. Porous collagen-based scaffolds have been utilized clinically to induce regeneration in skin and peripheral nerves, however still there is no complete explanation about the underlying mechanism. This thesis utilizes advanced microscopy to study the expression of contractile cell phenotypes during wound healing, a phenotype believed to affect significantly the final outcome. The first part develops an efficient pipeline for processing challenging spectral fluorescence microscopy images. Images are segmented into regions of objects by refining the outcome of a pixel-wide model selection classifier by an efficient Markov Random Field model. The methods of this part are utilized by the following parts. The second part extends the image informatics methodology in studying signal transduction networks in cells interacting with 3D matrices. The methodology is applied in a pilot study of TGFP signal transduction by the SMAD pathway in fibroblasts seeded in porous collagen scaffolds. Preliminary analysis suggests that the differential effect of TGFP1 and TGFP3 to cells could be attributed to the "non-canonical" SMADI and SMAD5. The third part is an ex vivo imaging study of peripheral nerve regeneration, which focuses on the formation of a capsule of contractile cells around transected rat sciatic nerves grafted with collagen scaffolds, 1 or 2 weeks post-injury. It follows a recent study that highlights an inverse relationship between the quality of the newly formed nerve tissue and the size of the contractile cell capsule 9 weeks post-injury. Results suggest that "active" biomaterials result in significantly thinner capsule already 1 week post-injury. The fourth part describes a novel method for quantifying the surface chemistry of 3D matrices. The method is an in situ binding assay that utilizes fluorescently labeled recombinant proteins that emulate the receptor of , and is applied to quantify the density of ligands for integrins a113, a2p1 on the surface of porous collagen scaffolds. Results provide estimates for the density of ligands on "active" and "inactive" scaffolds and demonstrate that chemical crosslinking can affect the surface chemistry of biomaterials, therefore can affect the way cells sense and respond to the material.by Dimitrios S. Tzeranis.Ph. D

The Grain of a Vocal Genre: A Comparative Approach to the Singing Pedagogies of EVDC Integrative Performance Practice, Korean Pansori, and the Centre for Theatre Practices 'Gardzienice'

This thesis is a cross-cultural examination of the relationship between the trained physiology of the voice and culture. Building on Barthes's notion of the grain of the voice,' I argue that each training system moulds the body in a way that decisively affects aesthetic phonation. Therefore I analyse voice training as a bodily inscription, in its Foucauldian sense, and I focus on the pedagogical ethics crystallised in the 'grain of the genre.' This I define as the collective 'grain' to which pedagogies of codified genres aspire, beyond and apart from the individual singing performer's 'grain' ; in other words, the 'grain of the genre' is the means by which culture is reaffirmed in/through the trainee's voice. The introductory chapter looks at the anatomical and physiological properties of the voice, traces the history of theorisations of the voice in order to situate my project, and explains my methodologies as a practitioner/researcher. Drawing on extended practical fieldwork, each of the subsequent three chapters explores the ' grain' of three pedagogies in the light of my personal training and the historical, musicological and broader cultural research I conducted in relation to each method. The three training approaches are a recent development in the area of bel canto (Integrative Performance Practice by 'Experience Vocal Dance Company,' USA and UK), an Asian traditional genre (Korean pansori), and a training pertaining to the Western avant-garde tradition (Centre for Theatre Practices 'Gardzienice,' Poland). Chapter 2 argues that the transdisciplinary grain of IPP on one hand adheres to a scientific approach to voicing, while attempting to bypass the deadends of the predominant training of the 'natural' voice. Chapter 3 acknowledges the grain of pansori as developed and promoted through the explicit aesthetic agenda of Korean han. Chapter 4 studies the grain of Gardzienice as one of 'laughing openness,' a grain mainly preoccupied with the inter-corporeal and the relational. The final chapter revisits the category of the 'grain of the genre' through my embodied perspective as a cross-cultural researcher. I reexamine the Foucauldian aspects of the 'grain' and its disciplinary character through the lens of the axis docility-resistance. I conclude the thesis with the suggestion of a dynamic relation between culture and vocal practice, the resistant aspects of which are, I argue, foregrounded when voice is addressed and taught as phonic and foreign.EThOS - Electronic Theses Online ServiceGBUnited Kingdo