Targeting the actin cytoskeleton in HER2+ metastatic cancer cells using a macrolide toxin

Breast and ovarian cancers are among the leading causes of cancer related deaths in women worldwide. In a subset of these cancers, dysregulation of the human epidermal growth factor receptor 2 (HER2) leads to overexpression of the receptor on the cell surface. Previous studies have found that these HER2+ cancers show high rates of progression to metastatic disease. Metastasis is driven by cytoskeletal rearrangements that produce filamentous actin (F-actin) based structures that penetrate and degrade extracellular matrix to facilitate tumour invasion. Advancements in targeted therapy have made F-actin an attractive target for the development of new cancer therapies. In this thesis, we tested the actin-depolymerizing macrolide toxin, Mycalolide B (MycB), as a potential warhead for a novel antibody drug conjugate (ADC) to target highly metastatic HER2+ breast and ovarian cancers. We found that MycB treatment of HER2+ breast (SKBR3, MDA-MB-453) and ovarian (SKOV3) cancer cells led to loss of viability (IC50 values ≤ 64 nM). Sub-lethal doses of MycB treatment caused potent suppression of leading edge protrusions, migration and invasion potential of HER2+ cancer cells (IC50 ≤ 32 nM). In contrast, other F-actin based processes such as receptor endocytosis were less sensitive to MycB treatment. MycB treatment skewed the size of endocytic vesicles, which may reflect defects in F-actin based vesicle motility or maturation. Given that HER2+ cancers have been effectively targeted by Trastuzumab and Trastuzumab-based ADCs, we tested the effects of a combination of Trastuzumab and MycB on cell migration and invasion. We found that MycB/ Trastuzumab combination treatments inhibited motility of SKOV3 cells to a greater degree than either treatment alone. Altogether, our results provide proof-of-principle that actin toxins such as MycB can be used as a novel class of warheads for ADCs to target and combat highly metastatic cancers.

The principle of legal certainty as a principle of economic efficiency

Legal certainty, a feature of the rule of law, constitutes a requirement for the operational necessities of market interactions. But, the compatibility of the principle of legal certainty with ideals such as liberalism and free market economy must not lead to the hastened conclusion that therefore the principle of legal certainty would be compatible and tantamount to the principle of economic efficiency. We intend to analyse the efficiency rationale of an important general principle of EU law—the principle of legal certainty. In this paper, we shall assert that not only does the EU legal certainty principle encapsulate an efficiency rationale, but most importantly, it has been interpreted by the ECJ as such. The economic perspective of the principle of legal certainty in the European context has, so far, never been adopted. Hence, we intend to fill in this gap and propose the representation of the principle of legal certainty as a principle of economic efficiency. After having deciphered the principle of legal certainty from a law and economics perspective (1), we shall delve into the jurisprudence of the ECJ so that the judicial reasoning of the Court as this reasoning proves the relevance of the proposed representation (2). Finally, we conclude in light of the findings of this paper (3).

Uguaglianza e differenza di genere nella riforma del diritto di famiglia: una lettura luhmanniana

La igualdad ante la ley es uno de los derechos fundamentales de las Constituciones modernas. Sin embargo, hay algo más. De hecho podría decirse que los modernos sistemas jurídicos se han ido formando alrededor de la idea de igualdad. El valor de la igualdad parece expresar una manera común de entender las relaciones humanas, especialmente en el caso de la igualdad de género, expresamente mencionado por el artículo de las Constituciones democráticas modernas que se refiere al principio de igualdad ante la ley. El presente estudio trata esa forma específica de igualdad que se caracteriza por la paradoja de querer tratar como igual lo que es diferente, o sea, de hacer distinciones sin discriminar. Con este fin nos referimos al enfoque jurídico y al enfoque político del tema como se encuentran en la reforma del derecho de familia. No es un problema de una específica legislación nacional, sino más bien una versión específica de un problema de nivel general. Se trata de la imposibilidad de construir como indistinguible lo que es diferente. El intento político de reformar el derecho de familia para implementar el principio constitucional de igualdad no consigue resolver el problema. Éste vuelve a aparecer por la imposibilidad que el derecho funcione sin crear diferencias. Al mismo tiempo, surgen problemas nuevos relacionado con formas nuevas de organizar la vida familiar.

Social media integration for the purpose of self-regulated learning

In this study, design based research in a teacher design team is performed. Teachers who intend to use social media to extend their school class teaching, are confronted with a lack of clear guidelines that inform them how to utilize them effectively. In this study, we aim at formulating such guidelines and testing their usefulness. First, opportunities to facilitate self-regulated learning (SRL) in higher education through the use of social media are elaborated in a literature study. Second, these opportunities are translated into teacher education to search for stimulating and limiting factors within the design, resulting in guidelines for effective social media integration.

A Scalable General Purpose System for Large-Scale Graph Processing

Graph analytics is an important and computationally demanding class of data analytics. It is essential to balance scalability, ease-of-use and high performance in large scale graph analytics. As such, it is necessary to hide the complexity of parallelism, data distribution and memory locality behind an abstract interface. The aim of this work is to build a scalable graph analytics framework that does not demand significant parallel programming experience based on NUMA-awareness.
The realization of such a system faces two key problems:
(i)~how to develop a scale-free parallel programming framework that scales efficiently across NUMA domains; (ii)~how to efficiently apply graph partitioning in order to create separate and largely independent work items that can be distributed among threads.

Multi-purpose utility of circulating plasma DNA testing in patients with advanced cancers.

Tumor genomic instability and selective treatment pressures result in clonal disease evolution; molecular stratification for molecularly targeted drug administration requires repeated access to tumor DNA. We hypothesized that circulating plasma DNA (cpDNA) in advanced cancer patients is largely derived from tumor, has prognostic utility, and can be utilized for multiplex tumor mutation sequencing when repeat biopsy is not feasible. We utilized the Sequenom MassArray System and OncoCarta panel for somatic mutation profiling. Matched samples, acquired from the same patient but at different time points were evaluated; these comprised formalin-fixed paraffin-embedded (FFPE) archival tumor tissue (primary and/or metastatic) and cpDNA. The feasibility, sensitivity, and specificity of this high-throughput, multiplex mutation detection approach was tested utilizing specimens acquired from 105 patients with solid tumors referred for participation in Phase I trials of molecularly targeted drugs. The median cpDNA concentration was 17 ng/ml (range: 0.5-1600); this was 3-fold higher than in healthy volunteers. Moreover, higher cpDNA concentrations associated with worse overall survival; there was an overall survival (OS) hazard ratio of 2.4 (95% CI 1.4, 4.2) for each 10-fold increase in cpDNA concentration and in multivariate analyses, cpDNA concentration, albumin, and performance status remained independent predictors of OS. These data suggest that plasma DNA in these cancer patients is largely derived from tumor. We also observed high detection concordance for critical 'hot-spot' mutations (KRAS, BRAF, PIK3CA) in matched cpDNA and archival tumor tissue, and important differences between archival tumor and cpDNA. This multiplex sequencing assay can be utilized to detect somatic mutations from plasma in advanced cancer patients, when safe repeat tumor biopsy is not feasible and genomic analysis of archival tumor is deemed insufficient. Overall, circulating nucleic acid biomarker studies have clinically important multi-purpose utility in advanced cancer patients and further studies to pursue their incorporation into the standard of care are warranted.

Fixed-boundary constituencies and the principle of equal representation in Ireland

The process of constituency boundary revision in Ireland, designed to satisfy what is perceived as a rigid requirement that a uniform deputy-population ratio be maintained across constituencies, has traditionally consumed a great deal of the time of politicians and officials. For almost two decades after a High Court ruling in 1961, the process was a political one, was highly contentious, and was marked by serious allegations of ministerial gerrymandering. The introduction in 1979 of constituency commissions made up of officials neutralised, for the most part, charges that the system had become too politicised, but it continued the process of micro-management of constituency boundaries. This article suggests that the continuing problems caused by this system – notably, the permanently changing nature of constituency boundaries and resulting difficulties of geographical identification – could be resolved by reversion to the procedure that is normal in proportional representation systems: periodic post-census allocation of seats to constituencies whose boundaries are based on those of recognised local government units and which are stable over time. This reform, replacing the principle of redistricting by the principle of reapportionment, would result in more recognisable constituencies, more predictable boundary trajectories over time, and a more efficient, fairer, and speedier process of revision.

Extending Automated Testing To High-level Software Requirements : A study on the feasibility of automated acceptance-testing

Automated acceptance testing is the testing of software done in higher level to test whether the system abides by the requirements desired by the business clients by the use of piece of script other than the software itself. This project is a study of the feasibility of acceptance tests written in Behavior Driven Development principle. The project includes an implementation part where automated accep- tance testing is written for Touch-point web application developed by Dewire (a software consultant company) for Telia (a telecom company) from the require- ments received from the customer (Telia). The automated acceptance testing is in Cucumber-Selenium framework which enforces Behavior Driven Development principles. The purpose of the implementation is to verify the practicability of this style of acceptance testing. From the completion of implementation, it was concluded that all the requirements from customer in real world can be converted into executable specifications and the process was not at all time-consuming or difficult for a low-experienced programmer like the author itself. The project also includes survey to measure the learnability and understandability of Gherkin- the language that Cucumber understands. The survey consist of some Gherkin exam- ples followed with questions that include making changes to the Gherkin exam- ples. Survey had 3 parts: first being easy, second medium and third most difficult. Survey also had a linear scale from 1 to 5 to rate the difficulty level for each part of the survey. 1 stood for very easy and 5 for very difficult. Time when the partic- ipants began the survey was also taken in order to calculate the total time taken by the participants to learn and answer the questions. Survey was taken by 18 of the employers of Dewire who had primary working role as one of the programmer, tester and project manager. In the result, tester and project manager were grouped as non-programmer. The survey concluded that it is very easy and quick to learn Gherkin. While the participants rated Gherkin as very easy.

National Office for Suicide Prevention annual report 2013

Suicide prevention is a significant public health issue in Ireland given the increase in suicide mortality and the emerging evidence of the negative impact of the economic downturn on mental health. In 2013, work commenced on the development of a new National Strategic Framework for Suicide Prevention. This Framework will provide a clear road map in relation to suicide prevention in Ireland. It will build upon the valuable work completed under Reach Out, the current National Strategy, and will reflect the best national and international evidence on suicide prevention to provide a clear set of actions and outcomes. The key principle to the Framework development is that of working together with our partners and the community, with a sense of common purpose, to achieve our shared aim: reducing the number of suicides in Ireland.   .

Imperfect indifference : the rhythm, structure and politics of neutrality

Cette thèse propose l’émergence d’une poésie de l’entre deux dans la littérature expérimentale, en suivant ses développements du milieu du vingtième siècle jusqu'au début du vingt-et-unième. Cette notion d’entre-deux poétique se fonde sur une théorie du neutre (Barthes, Blanchot) comme ce qui se situe au delà ou entre l'opposition et la médiation. Le premier chapitre retrace le concept de monotonie dans la théorie esthétique depuis la période romantique où il est vu comme l'antithèse de la variabilité ou tension poétique, jusqu’à l’émergence de l’art conceptuel au vingtième siècle où il se déploie sans interruption. Ce chapitre examine alors la relation de la monotonie à la mélancolie à travers l’analyse de « The Anatomy of Monotony », poème de Wallace Stevens tiré du recueil Harmonium et l’œuvre poétique alphabet de Inger Christensen. Le deuxième chapitre aborde la réalisation d’une poésie de l’entre-deux à travers une analyse de quatre œuvres poétiques qui revisitent l’usage de l’index du livre paratextuel: l’index au long poème “A” de Louis Zukofsky, « Index to Shelley's Death » d’Alan Halsey qui apparait à la fin de l’oeuvre The Text of Shelley's Death, Cinema of the Present de Lisa Robertson, et l’oeuvre multimédia Via de Carolyn Bergvall. Le troisième chapitre retrace la politique de neutralité dans la théorie de la traduction. Face à la logique oppositionnelle de l’original contre la traduction, il propose hypothétiquement la réalisation d’une troisième texte ou « l’entre-deux », qui sert aussi à perturber les récits familiers de l’appropriation, l’absorption et l’assimilation qui effacent la différence du sujet de l’écrit. Il examine l’oeuvre hybride Secession with Insecession de Chus Pato et Erin Moure comme un exemple de poésie de l’entre-deux. A la fois pour Maurice Blanchot et Roland Barthes, le neutre représente un troisième terme potentiel qui défie le paradigme de la pensée oppositionnelle. Pour Blanchot, le neutre est la différence amenée au point de l’indifférence et de l’opacité de la transparence tandis que le désire de Barthes pour le neutre est une utopie lyrique qui se situe au-delà des contraintes de but et de marquage. La conclusion examine comment le neutre correspond au conditions de liberté gouvernant le principe de créativité de la poésie comme l’acte de faire sans intention ni raison.

Êtres et avoirs- Esquisse sur les droits sans sujet en droit privé

En 1994, le législateur québécois a transformé la fiducie en un patrimoine d’affectation, c’est-à-dire un ensemble de biens et d’obligations sans titulaire, administré par un tiers, pour une fin particulière. Cette nouvelle qualification remet en question l’assise classique du droit privé : le rapport inhérent entre un droit et son titulaire, le sujet de droit, dont le droit émane et pour qui il est, en principe, exercé. Cette thèse tente de comprendre quelles sont les conditions de possibilités des droits sans sujet en droit civil actuel. Mon objectif est de repenser la notion de droit subjectif de manière générale afin qu’elle prenne en considération l’existence et la pertinence de ces droits sans sujet. Pour ce faire, deux entreprises sont nécessaires : (i) comprendre le paradigme dominant afin de bien cerner l’ampleur de la nouveauté et la résistance qu’elle engendre ; (ii) mettre au jour les fondements et les limites des droits sans sujet afin d’évaluer, dans une prochaine étape, si cette manière de comprendre les droits ne serait pas une alternative intéressante aux droits subjectifs actuels.

Urbaania energiaa. Suvilahden, Hanasaaren A- ja Vuosaaren voimalaitokset osana helsinkiläistä kaupunkiympäristöä

Tutkimus koostuu kolmen Helsingissä sijaitsevan, kolmena eri aikakautena, 1910-, 1950- ja 1990-luvuilla, rakennetun voimalaitoksen arkkitehtuurista ja rakennustyypistä sekä niiden eroista ja erityisyyksistä samoin kuin näiden voimalaitosten roolista ja vaikutuksesta Helsingin kaupunkisuunnitteluun ja -rakentamiseen, kaupunkikuvaan sekä ympäristöestetiikkaan. Tutkimus on rajattu koskemaan erityyppisten voimalaitosten osalta yksinomaan kolmea helsinkiläistä kaupungissa sijaitsevaa voimalaitosta, Suvilahtea, Hanasaari-A:ta ja Vuosaaren A- ja B-laitoksia. Tutkimuksen tarkoituksena on ensinnäkin selvittää sekä periaatteessa että edellä mainittujen kolmen esimerkkikohteen kautta seikkoja, jotka ovat vaikuttaneet kunkin voimalaitoksen arkkitehtuuriin ja rakennustyyppiin kunakin aikakautena. Kaupunkivoimalan olennaiset elementit ovat korkea savupiippu, mittava polttoainevarasto sekä massiiviset rakennusmassat, jotka vaativat runsasta maankäyttöä. Toiseksi tutkimuksessa paneudutaan kaupunkisuunnitteluun laitoksen sijoittumisen osalta sekä ajallisesti että paikallisesti. Kolmanneksi selvitetään kaupunkikuvallisia ja ympäristöesteettisiä seikkoja, sekä niiden vaikutusten kehitystä voimalaitoksen toteutuksen ja nykyhetken kesken. Tutkimuksessa haetaan vastausta kysymykseen, miten Helsingissä sijaitseva voimalaitos arkkitehtuuriltaan, rakennustyypiltään ja sijoitukseltaan on soveltunut ja jatkossa soveltuu kaupunkisuunnittelun kannalta kaupunkikuvallisesti sekä ympäristöesteettisesti kyseiseen kaupunkiympäristöön. Tutkimus selvittää myös sitä ilmeistä ristiriitaa, joka syntyy kaupungin kehittyessä ja laajentuessa, jolloin voimalaitos infrastruktuurinsa ja useimmiten suunnattoman kokonsa vuoksi edustaa pysyvyyttä rakentuvan alueen sisällä. Tässä yhteydessä tutkimuksessa pohditaan esimerkkikohteiden avulla voimalaitoksen säilyttämistä puoltavia rakennustaiteellisia arvoja, mahdollista korvattavuutta, ja siinä yhteydessä haetaan vastauksia jäljelle jäävälle laitosrakennukselle kaupunkisuunnittelun kannalta asetettavista uusiokäytön vaatimuksista ja mahdollisuuksista. Tutkimuksen metodologia on sekä kvantitatiivisesti että kvalitatiivisesti historiallinen, esimerkkikohteita käsiteltäessä tarvittavassa määrin myös mikrohistoriallinen.