The higher pragmitism

Thesis (M.A.)--Boston Universit

Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors.

Lentivirus-based gene delivery vectors carrying multiple gene cassettes are powerful tools in gene transfer studies and gene therapy, allowing coexpression of multiple therapeutic factors and, if desired, fluorescent reporters. Current strategies to express transgenes and microRNA (miRNA) clusters from a single vector have certain limitations that affect transgene expression levels and/or vector titers. In this study, we describe a novel vector design that facilitates combined expression of therapeutic RNA- and protein-based antiangiogenic factors as well as a fluorescent reporter from back-to-back RNApolII-driven expression cassettes. This configuration allows effective production of intron-embedded miRNAs that are released upon transduction of target cells. Exploiting such multigenic lentiviral vectors, we demonstrate robust miRNA-directed downregulation of vascular endothelial growth factor (VEGF) expression, leading to reduced angiogenesis, and parallel impairment of angiogenic pathways by codelivering the gene encoding pigment epithelium-derived factor (PEDF). Notably, subretinal injections of lentiviral vectors reveal efficient retinal pigment epithelium-specific gene expression driven by the VMD2 promoter, verifying that multigenic lentiviral vectors can be produced with high titers sufficient for in vivo applications. Altogether, our results suggest the potential applicability of combined miRNA- and protein-encoding lentiviral vectors in antiangiogenic gene therapy, including new combination therapies for amelioration of age-related macular degeneration

Growing blood vessels in space : preparation studies of the SPHEROIDS project using related ground-based studies

Endothelial cells (ECs) grow as single layers on the bottom surface of cell culture flasks under normal (1g) culture conditions. In numerous experiments using simulated microgravity we noticed that the ECs formed three-dimensional, tube-like cell aggregates resembling the intima of small, rudimentary blood vessels. The SPHEROIDS project has now shown that similar processes occur in space. For the first time, we were able to observe scaffold-free growth of human ECs into multicellular spheroids and tubular structures during an experiment in real microgravity. With further investigation of the space samples we hope to understand endothelial 3D growth and to improve the in vitro engineering of biocompatible vessels which could be used in surgery

Unexpected Hydrolytic Instability of N-Acylated Amino Acid Amides and Peptides

Remote amide bonds in simple N-acyl amino acid amide or peptide derivatives 1 can be surprisingly unstable hydrolytically, affording, in solution, variable amounts of 3 under mild acidic conditions, such as trifluoroacetic acid/water mixtures at room temperature. This observation has important implications for the synthesis of this class of compounds, which includes N-terminal-acylated peptides. We describe the factors contributing to this instability and how to predict and control it. The instability is a function of the remote acyl group, R2CO, four bonds away from the site of hydrolysis. Electron-rich acyl R2 groups accelerate this reaction. In the case of acyl groups derived from substituted aromatic carboxylic acids, the acceleration is predictable from the substituent’s Hammett σ value. N-Acyl dipeptides are also hydrolyzed under typical cleavage conditions. This suggests that unwanted peptide truncation may occur during synthesis or prolonged standing in solution when dipeptides or longer peptides are acylated on the N-terminus with electron-rich aromatic groups. When amide hydrolysis is an undesired secondary reaction, as can be the case in the trifluoroacetic acid-catalyzed cleavage of amino acid amide or peptide derivatives 1 from solid-phase resins, conditions are provided to minimize that hydrolysis

The Ontological Significance of Consciousness

Providing a wholly physical description of consciousness in nature is an elusive process. Unlike all other physical systems, consciousness seems to be explainable only if it is considered as non-physical. However, the arguments for a materialist view of consciousness, with consciousness prioritized as a process which creates imperfect dependence relations in reality, eliminates the need for a dualist understanding of reality. Through an explanation of imperfect dependence, materialist consciousness overcomes all relevant dualist arguments, while simultaneously retaining an epistemic gap, denying the conceivability of zombies, and providing convincing explanations of seemingly dualist positions in a materialist fashion

Growth of endothelial cells in space and in simulated microgravity : a comparison on the secretory level

BACKGROUND/AIMS: Endothelial cells exposed to the Random Positioning Machine (RPM) reveal three different phenotypes. They grow as a two-dimensional monolayer and form three-dimensional (3D) structures such as spheroids and tubular constructs. As part of the ESA-SPHEROIDS project we want to understand how endothelial cells (ECs) react and adapt to long-term microgravity. METHODS: During a spaceflight to the International Space Station (ISS) and a subsequent stay onboard, human ECs (EA.hy926 cell line) were cultured for 12 days in real microgravity inside an automatic flight hardware, specially designed for use in space. ECs were cultivated in the absence or presence of vascular endothelial growth factor, which had demonstrated a cell-protective effect on ECs exposed to an RPM simulating microgravity. After cell fixation in space and return of the samples, we examined cell morphology and analyzed supernatants by Multianalyte Profiling technology. RESULTS: The fixed samples comprised 3D multicellular spheroids and tube-like structures in addition to monolayer cells, which are exclusively observed during growth under Earth gravity (1g). Within the 3D aggregates we detected enhanced collagen and laminin. The supernatant analysis unveiled alterations in secretion of several growth factors, cytokines, and extracellular matrix components as compared to cells cultivated at 1g or on the RPM. This confirmed an influence of gravity on interacting key proteins and genes and demonstrated a flight hardware impact on the endothelial secretome. CONCLUSION: Since formation of tube-like aggregates was observed only on the RPM and during spaceflight, we conclude that microgravity may be the major cause for ECs' 3D aggregation.status: publishe

Motivating and engaging high school students in citizen science projects

Citizen science (CS) in education programs at high schools has the aim to engage youth in research with a hands-on, active learning process. Therefore, the SDU Citizen Science Center has participated as a practitioner in several CS projects within social sciences, natural science and humanities that engage Danish high school students as citizen scientists as part of their education.In “Climate Future Fiction” the students contribute to research on young people’s concerns regarding climate change in the far-future by writing and analyzing Cli-fi stories in their English classes. In “Our History” the students participate as citizen scientists by interviewing elderly people about their experiences of the societal changes in Denmark in the 1960s and 1970s, thereby creating data for the researchers as part of their history lessons. “A Healthier Southern Denmark” is a project and competition with five researchers from the region hospitals, in parallel a “High School Panel” is established. As part of their social science lessons, the students are in dialogue with the researchers on how to disseminate research to citizens.Some 74 Danish high school classes have participated in these projects since 2019. Experiences from these CS projects are the focus of this poster:• A model on how to engage high school students in CS.• The perspective of engaging the students in CSIn co-creation with high school teachers and researchers, we have developed a semi-generic model for implementing CS in high school education. The model includes a masterclass for the teachers, co-creation of curriculum-based learning materials, dialogue between the researchers and the high school students and a competition element for the students with feedback from the researchers. Evaluations and follow-up research shows that the participating students increase their scientific knowledge, hopefully, this will change their attitudes towards science in the long run

Motivating and engaging high school students in citizen science projects

Citizen science (CS) in education programs at high schools has the aim to engage youth in research with a hands-on, active learning process. Therefore, the SDU Citizen Science Center has participated as a practitioner in several CS projects within social sciences, natural science and humanities that engage Danish high school students as citizen scientists as part of their education.In “Climate Future Fiction” the students contribute to research on young people’s concerns regarding climate change in the far-future by writing and analyzing Cli-fi stories in their English classes. In “Our History” the students participate as citizen scientists by interviewing elderly people about their experiences of the societal changes in Denmark in the 1960s and 1970s, thereby creating data for the researchers as part of their history lessons. “A Healthier Southern Denmark” is a project and competition with five researchers from the region hospitals, in parallel a “High School Panel” is established. As part of their social science lessons, the students are in dialogue with the researchers on how to disseminate research to citizens.Some 74 Danish high school classes have participated in these projects since 2019. Experiences from these CS projects are the focus of this poster:• A model on how to engage high school students in CS.• The perspective of engaging the students in CSIn co-creation with high school teachers and researchers, we have developed a semi-generic model for implementing CS in high school education. The model includes a masterclass for the teachers, co-creation of curriculum-based learning materials, dialogue between the researchers and the high school students and a competition element for the students with feedback from the researchers. Evaluations and follow-up research shows that the participating students increase their scientific knowledge, hopefully, this will change their attitudes towards science in the long run