Search results for "EXPRESSION"
showing 10 items of 5168 documents
Persistent episomal transgene expression in liver following delivery of a scaffold/matrix attachment region containing non-viral vector
2008
An ideal gene therapy vector should enable persistent transgene expression without limitations of safety and reproducibility. Here we report the development of a non-viral episomal plasmid DNA (pDNA) vector that appears to fulfil these criteria. This pDNA vector combines a scaffold/matrix attachment region (S/MAR) with a human liver-specific promoter (alpha1-antitrypsin (AAT)) in such a way that long-term expression is enabled in murine liver following hydrodynamic injection. Long-term expression is demonstrated by monitoring the longitudinal luciferase expression profile for up to 6 months by means of in situ bioluminescent imaging. All relevant control pDNA constructs expressing luciferas…
Oxidative stress responses and lipid peroxidation damage are induced during dehydration in the production of dry active wine yeasts.
2009
The tolerance of the yeast Saccharomyces cerevisiae to desiccation is important for the use of this microorganism in the wine industry, since active dry wine yeast is routinely used as starter for must fermentations. Many studies have shown the complexity of the cellular effects caused by water loss, including oxidative injuries on macromolecular components. However the technological interest of yeast drying was not addressed in those studies, and the dehydration conditions were far from the industrial practice. In the present study a molecular approach was used to characterize the relevant injuring conditions during pilot plant dehydrations under two different drying temperatures (i.e., 35…
D2R striatopallidal neurons inhibit both locomotor and drug reward processes.
2009
The specific functions of dopamine D(2) receptor-positive (D(2)R) striatopallidal neurons remain poorly understood. Using a genetic mouse model, we found that ablation of D(2)R neurons in the entire striatum induced hyperlocomotion, whereas ablation in the ventral striatum increased amphetamine conditioned place preference. Thus D(2)R striatopallidal neurons limit both locomotion and, unexpectedly, drug reinforcement.
Hermeneutics of Transcendence. Understanding and Communication at the Limits of Experience
2013
When access to shared realities is denied or blocked as in extreme situations that transcend everyday experience, a phenomenological-hermeneutic problem arises between the solitary Ego and its socially constituted meaning. With no action space to prove one’s own reality there lacks a counter-space to transcend the ambiguously irreal reality in order to perspectivize, to understand and reflect on it. Hermeneutics of transcendence analyse the experience of borders and limitations – not starting with an interrogation of constitutive grounds of shared social meaning but with structures to regain meaning in the socio-pathological structures of life excluding a Self from horizons of shared meanin…
The Qualitative Researcher in the Quality of Life Field
2015
In the current society, there is a plurality of perspectives about the knowledge of the world and in that stage there is an expansion of the use of qualitative methods as expression of the contemporary interest in the subject-matters, considering the epistemological relevance acquired by the introduction of the subject’s word as a possibility to construct knowledge. The qualitative researcher is expected to feel personally involved in every step of the research process, because every consideration and decision will have to be based on entirely personal grounds. Thus, in this chapter, we shall look into the importance of the qualitative researcher’s leading role in each phase of the research…
A complete set of nascent transcription rates for yeast genes
2010
The amount of mRNA in a cell is the result of two opposite reactions: transcription and mRNA degradation. These reactions are governed by kinetics laws, and the most regulated step for many genes is the transcription rate. The transcription rate, which is assumed to be exercised mainly at the RNA polymerase recruitment level, can be calculated using the RNA polymerase densities determined either by run-on or immunoprecipitation using specific antibodies. The yeast Saccharomyces cerevisiae is the ideal model organism to generate a complete set of nascent transcription rates that will prove useful for many gene regulation studies. By combining genomic data from both the GRO (Genomic Run-on) a…
Genome-wide studies of mRNA synthesis and degradation in eukaryotes
2012
In recent years, the use of genome-wide technologies has revolutionized the study of eukaryotic transcription producing results for thousands of genes at every step of mRNA life. The statistical analyses of the results for a single condition, different conditions, different transcription stages, or even between different techniques, is outlining a totally new landscape of the eukaryotic transcription process. Although most studies have been conducted in the yeast Saccharomyces cerevisiae as a model cell, others have also focused on higher eukaryotes, which can also be comparatively analyzed. The picture which emerges is that transcription is a more variable process than initially suspected,…
Transcriptional targeting of dendritic cells for gene therapy using the promoter of the cytoskeletal protein fascin.
2003
Strong cell-type-specific promoters are basic tools in gene therapy allowing for novel applications and focused strategies by transcriptionally targeting gene expression to selected cells. In immunotherapy, dendritic cells (DC) are of central importance, since they represent the principal inducers of immune responses. Here we describe isolation and use of the promoter of the murine actin-bundling protein fascin to target transcriptionally gene expression to cutaneous DC. Using the reporter gene enhanced green fluorescent protein (EGFP), we demonstrate that the fascin promoter mediates a strong antigen expression that is restricted to mature DC. DNA vaccination with antigen-encoding expressi…
Stability of phospholipase D in primary astrocytes.
2002
Induction of expression and proteolytic breakdown of phospholipase D (PLD) isoforms in primary astrocyte cultures have been investigated. Astrocytes express both PLD1 and 2 and are dependent on PLD activity for cell proliferation [K. Kotter, J. Klein, J. Neurochem. 73 (1999) 2517]. Competitive RT-PCR analysis demonstrated a higher level of PLD1 mRNA than PLD2 mRNA (8.9 vs. 0.9amol/microg RNA, respectively). Treatment of astroglial cultures with the phorbol ester, 4beta-phorbol-12beta,13alpha-dibutyrate (0.1 microM), for 24-48h selectively induced PLD1b but not PLD1a or 2 expression as shown by PCR and Western blot; the effect was sensitive to Go 6976. In cells transiently permeabilized with…
Molecular characterization of an inducible p-coumaric acid decarboxylase from Lactobacillus plantarum: gene cloning, transcriptional analysis, overex…
1997
By using degenerate primers designed from the first 19 N-terminal amino acids of Lactobacillus plantarum p-coumaric acid decarboxylase (PDC), a 56-bp fragment was amplified from L. plantarum in PCRs and used as a probe for screening an L. plantarum genomic bank. Of the 2,880 clones in the genomic bank, one was isolated by colony hybridization and contained a 519-bp open reading frame (pdc gene) followed by a putative terminator structure. The pdc gene is expressed on a monocistronic transcriptional unit, which is transcribed from promoter sequences homologous to Lactococcus promoter sequences. No mRNA from pdc and no PDC activity were detected in uninduced cell extracts, indicating that the…