Mesenchymale Stromazellen der nächsten Generation für regenerative Zwecke - Manipulation des Zellphänotyps durch Biomaterialien und nichtviralen Gentransfer

The sum of different microenvironment stimuli precisely regulates cell phenotype and behavior. Synergistic strategies, which make use of this multimodality could be harnessed to precisely control functions of therapeutic cells. In this thesis I have established and applied a combination of biomaterial encapsulation and mRNA transfection to shape the paracrine profile of mesenchymal stromal cells (MSCs), a secretory active cell type shown to be safe but not always efficacious in clinical trials. To stimulate growth factor secretion through cell-material interactions, MSCs were encapsulated in collagen and HA (Col-HA) hydrogel, or control hydrogels made from HA, PEG, and gelatin. MSC showed stronger elongation and an expanded growth factor profile in Col-HA with distinct upregulation kinetics for HGF versus VEGF. MSCs were transfected with in vitro transcribed mRNA coding for GFP and VEGF using lipofectamine. Uridine was substituted by different chemically modified derivates, including 5-methoxy-uridine (5moU). Unmodified mRNA triggered a type-I interferon response in MSCs and upregulated pro-inflammatory cytokine secretion, while downregulating pro-angiogenic growth factors. 5moU-mRNA did not trigger a detectable type-I interferon response and allowed for prolonged gene overexpression without disturbing the MSC's natural secretome. Finally, a protocol for sequential mRNA transfection and hydrogel encapsulation was developed. The effects of mRNA transfection on cell-material interactions were evaluated. Only 5moU-mRNA, but not unmodified mRNA, allowed for sequential transfection and encapsulation with high cell viability and preserved hydrogel-dependent effects on growth factor secretion. Conversely, the effect of cell-matrix adhesion on mRNA transfection was investigated by comparing GFP expression and VEGF secretion in Col-HA, which contains integrin ligand motifs, versus a PEG-HA control gel, which does not. In Col-HA, MSCs secreted higher levels of mRNA-encoded VEGF than in PEG-HA, but no difference in GFP fluorescence was observed, suggesting that secretion, but not translation of mRNA-encoded protein is adhesion-sensitive. The results show that broad secretome stimulation by cell-material interactions can be orthogonally combined with high mRNA overexpression of a selected key factor, to shape the MSC secretome on multiple levels. Additionally, we identified a synergy be-tween secretion of targeted mRNA-encoded factors and cell adhesion. Importantly, 5moU-mRNA can be used to express a gene of interest while leaving the remaining cellular phenotype largely unaffected. Open questions include elucidating mechanistic connections between cell-material interactions and the temporal changes in secretion profile, as well as showing a therapeutic utility of such a multimodal system. The concept proposed here could be expanded into generalized stimulome models of synergies and antagonisms between different cell-instructive stimuli.Die Summe verschiedener Reize aus dem Mikromilieu reguliert den Phänotyp und das Verhalten von Zellen. Synergistische Strategien, welche diese Multimodalität berücksich tigen, könnten genutzt werden, um die Funktion therapeutischer Zellen präzise zu kon trollieren. In dieser Arbeit habe ich eine Kombination aus Biomaterialeinbettung und mRNA Transfektion etabliert, um das parakrine Profil Mesenchymaler Stromazellen (MSCs) zu formen. MSCs sind ein sekretorisch aktiver Zelltyp, welcher sich in klinischen Studien als sicher, aber nicht immer wirkungsvoll erwiesen hat. Um Wachstumsfaktorsekretion mittels Zell-Material-Interaktionen zu stimulieren, wurden MSCs in Kollagen-HA (Col-HA) Hydrogel eingebettet. Als Kontrolle dienten HA-, PEG und Gelatinegele. MSCs in Col-HA wiesen stärkere Elongation und ein expandiertes Sek retionsprofil mit unterschiedlichen Kinetiken für HGF und VEGF auf. MSCs wurden mittels Lipofektamin mit in vitro transkribierter mRNA transfiziert, welche GFP bzw. VEGF kodiert. Uridin wurde durch Varianten wie 5-Methoxy-Uridin (5moU) er setzt. Unmodifizierte mRNA löste eine Typ-I Interferonantwort aus, erhöhte die Ausschüt tung proinflammatorischer Zytokine und senkte proangiogene Faktoren. 5moU-mRNA löste keine detektierbare Interferonantwort aus und erlaubte eine verlängerte Genexpres sion, ohne Beeinträchtigung des natürlichen Sekretionsprofils. Zuletzt wurde die mRNA Transfektion sequentiell mit der Hydrogeleinbettung kombiniert. Nur 5moU-mRNA, nicht jedoch unmodifizierte mRNA erlaubte die sequentielle Kombina tion beider Methoden bei hoher Zellviabilität und erhaltenen Hydrogeleffekten auf die Fak torfreisetzung. Umgekehrt wurde auch der Einfluss der Matrixadhäsion auf die mRNA Transfektion untersucht. GFP Expression und VEGF Sekretion wurden in Col-HA versus PEG-HA, welches keine Integrinliganden enthält, verglichen. In Col-HA sezernierten die MSCs mehr mRNA-kodiertes VEGF als in PEG-HA. Das GFP Signal war vergleichbar, was nahelegt, dass nicht die Translation, sondern die Sekretion adhäsionssensitiv ist. Die Ergebnisse zeigen, dass die breite Sekretomstimulation durch Zell-Material-Interak tionen orthogonal mit mRNA Überexpression eines ausgewählten Schlüsselmediators kombiniert werden kann, um das MSC Sekretom auf meheren Ebenen zu formen. 5moU-mRNA kann zur Expression eines gewünschten Gens verwendet werden, ohne den übrigen Zellphänotyp zu beeinträchtigen. Zudem konnten wir einen Synergismus zwischen Zelladhäsion und der Freisetzung mRNA-kodierter Faktoren identifizieren. Offen bleibt die Frage nach den genauen Mechanismen, durch welche die Zell-Material-Interaktionen zu zeitlichen Veränderungen im Sekretionsprofil führen. Außerdem muss der therapeutische Nutzen eines solchen multimodalen Systems gezeigt werden. Das hier vorgeschlagene Konzept könnte zu allgemeineren Stimulom-Modellen synergistischer bzw. antagonistischer Reizinteraktionen erweitert werden

The Inclusive Growth and Development Report 2017

Around the globe, leaders of governments and other stakeholder institutions enter 2017 facing a set of difficult and increasingly urgent questions:With fiscal space limited, interest rates near zero, and demographic trends unfavorable in many countries, does the world economy face a protracted period of relatively low growth? Will macroeconomics and demography determine the world economy's destiny for the foreseeable future?Can rising in-country inequality be satisfactorily redressed within the prevailing liberal international economic order? Can those who argue that modern capitalist economies face inherent limitations in this regard – that their internal "income distribution system" is broken and likely beyond repair – be proven wrong?As technological disruption accelerates in the Fourth Industrial Revolution, how can societies organize themselves better to respond to the potential employment and other distributional effects? Are expanded transfer payments the only or primary solution, or can market mechanisms be developed to widen social participation in new forms of economic value-creation?These questions beg the more fundamental one of whether a secular correction is required in the existing economic growth model in order to counteract secular stagnation and dispersion (chronic low growth and rising inequality). Does the mental map of how policymakers conceptualize and enable national economic performance need to be redrawn? Is there a structural way, beyond the temporary monetary and fiscal measures of recent years, to cut the Gordian knot of slow growth and rising inequality, to turn the current vicious cycle of stagnation and dispersion into a virtuous one in which greater social inclusion and stronger and more sustainable growth reinforce each other?This is precisely what government, business, and other leaders from every region have been calling for. Over the past several years, a worldwide consensus has emerged on the need for a more inclusive growth and development model; however, this consensus is mainly directional. Inclusive growth remains more a discussion topic than an action agenda. This Report seeks to help countries and the wider international community practice inclusive growth and development by offering a new policy framework and corresponding set of policy and performance indicators for this purpose

Оптимизация применения технологических составов для обработки скважин на нефтяном месторождении "Х" (Томская область)

Цель исследования – анализ методов и технологий химической защиты внутрискважинного оборудования от осложнений на месторождении "Х". Объектом исследования являются осложнения, возникающие при эксплуатации нефтяных скважин и химические методы их предотвращения. В процессе исследования была подробно рассмотрена классификация современных рецептур (по действующему веществу) ингибиторов солеотложений, а также перспективы по применению более совершенных реагентов. Проведен анализ технологий и сопутствующих им химических реагентов, применяющихся для борьбы с солеотложениями и коррозией на месторождении "Х". Проанализированы современные тренды при организации ингибиторной защиты от солеотложений и коррозии. В результате исследования выявлен положительный эффект ингибирования скважин капсулированныThe purpose of the study is to analyze the methods and technologies of chemical protection of downhole equipment from complications at the “X” field. The object of research is the complications arising from the operation of oil wells and chemical methods to prevent them. In the course of the study, the classification of modern formulations (according to the active substance) of scale inhibitors, as well as prospects for the use of more sophisticated reagents, were reviewed in detail. The analysis of technologies and related chemical reagents used to combat scaling and corrosion in the field "X". Analyzed modern trends in the organization of inhibitor protection against scaling and corrosion. The study revealed a positive effect of inhibition of wells with encapsulated reagents and reagent

Комплексная эколого-геохимическая оценка состояния природной среды на территории г. Горняк (Алтайский край)

Объектом исследования является территория г. Горняк (Алтайский край). Предметом исследования являются компоненты природной среды (волосы, накипь, почва, материал хвостохранилищ и листья тополя). Цель работы – изучить элементный состав природной среды и организма человека на территории г. Горняк (Алтайский край).The object of the study is the territory of the town of Gornyak (Altai Territory). The subject of the study is the components of the natural environment (hair, scale, soil, tailings material and poplar leaves). The purpose of the work is to study the elemental composition of the natural environment and the human body in the city of Gornyak (Altai Territory)

Hemispheric reconfigurations in Northern Amazonia: The ‘Three Guianas’ amid regional change and Brazilian hegemony

© 2016 Southseries Inc., www.thirdworldquarterly.com. Regional and hemispheric reconfigurations in Latin America and the Caribbean are increasingly mediated by Brazilian power, and the engagement of Guyana, Suriname and French Guiana with this emerging context is intriguing. They are tentatively moving away from a Caribbean region with which they are culturally contiguous, towards a South American continent in which they are geographically located. This is partly a reflection of the gradual opening up of the Northern Amazonian space that they share collectively, and also with Venezuela and Brazil. These processes are occurring as cause and effect of Brazil’s emergence as a regional–and even regionally hegemonic–power. With reference to wider debates on regionalism and hegemony, we analyse the uncertain consequences of these shifts