Protein purification and cDNA cloning of suGF1 : a sea urchin nuclear DNA-binding factor

The upstream regulatory regions of numerous genes contain contiguous deoxyguanosine residues (G·C-rich sequences) which have been implicated in the regulation of gene expression, since they may involve alterations in their DNA structure, the binding of G-string factors and in some cases even the displacement of a nucleosome positioned over this area. A poly( dG).poly( de)-binding protein (suGF1) has previously been identified and purified on a small scale from embryonic nuclear extracts of the sea urchin Parechinus angulosus (1, 2). suGF1 binds in vitro to the H1-H4 intergenic region of the early histone gene battery, and the recognition site contains 11 contiguous Gs which are incorporated into a positioned nucleosome core in vitro. suGFI may be a member of a family of Gstring factors which could be involved in the developmental regulation of unrelated genes in various organisms. Prior to the commencement of this project no protein or DNA sequence information was available on the protein. The main objective of this thesis was to obtain the eDNA and the primary amino acid sequence for suGFI. Using this information, additional aims were to determine the developmental distribution of the protein and obtain insight into the molecular basis of the regulatory function of suGF 1 by analysis of the primary protein structure and expression of the eDNA

Incorporation of aptamers in the terminal loop of shRNAs yields an effective and novel combinatorial targeting strategy.

Gene therapy by engineering patient's own blood cells to confer HIV resistance can potentially lead to a functional cure for AIDS. Toward this goal, we have previously developed an anti-HIV lentivirus vector that deploys a combination of shRNA, ribozyme and RNA decoy. To further improve this therapeutic vector against viral escape, we sought an additional reagent to target HIV integrase. Here, we report the development of a new strategy for selection and expression of aptamer for gene therapy. We developed a SELEX protocol (multi-tag SELEX) for selecting RNA aptamers against proteins with low solubility or stability, such as integrase. More importantly, we expressed these aptamers in vivo by incorporating them in the terminal loop of shRNAs. This novel strategy allowed efficient expression of the shRNA-aptamer fusions that targeted RNAs and proteins simultaneously. Expressed shRNA-aptamer fusions targeting HIV integrase or reverse transcriptase inhibited HIV replication in cell cultures. Viral inhibition was further enhanced by combining an anti-integrase aptamer with an anti-HIV Tat-Rev shRNA. This construct exhibited efficacy comparable to that of integrase inhibitor Raltegravir. Our strategy for the selection and expression of RNA aptamers can potentially extend to other gene therapy applications

A life in secret: aspects of the reception

This paper aims to present an analysis of the film "A secret life", an adaptation of the homonymous work of Autran Dourado. Intercrossing issues related to the perspective of Reception Studies from the strategies that are used, and the effects they produce in the viewer, the analysis allows a reflection on how stigma interferes in social roles, as shown in the relationship between the character Biela and others that integrate the narrative. Organized in stages, this study first presents some theoretical considerations about reading and reception and discusses the strategies deployed in these processes. Shortly after, we also describe the key scenes of the film followed by the analysis

Ensuring connectedness for the Maximum Quasi-clique and Densest kk-subgraph problems

Given an undirected graph $G$, a quasi-clique is a subgraph of $G$ whose density is at least $\gamma$ $(0 < \gamma \leq 1)$. Two optimization problems can be defined for quasi-cliques: the Maximum Quasi-Clique (MQC) Problem, which finds a quasi-clique with maximum vertex cardinality, and the Densest $k$-Subgraph (DKS) Problem, which finds the densest subgraph given a fixed cardinality constraint. Most existing approaches to solve both problems often disregard the requirement of connectedness, which may lead to solutions containing isolated components that are meaningless for many real-life applications. To address this issue, we propose two flow-based connectedness constraints to be integrated into known Mixed-Integer Linear Programming (MILP) formulations for either MQC or DKS problems. We compare the performance of MILP formulations enhanced with our connectedness constraints in terms of both running time and number of solved instances against existing approaches that ensure quasi-clique connectedness. Experimental results demonstrate that our constraints are quite competitive, making them valuable for practical applications requiring connectedness

Bilaterale III

Mit einem konkreten Vorschlag für ein umfassendes Bilaterale-III-Paket kann die Schweiz das Verhandlungsresultat positiv beeinflussen: Die Autorenschaft vertieft einen entsprechenden Vorschlag, der aktuell diskutiert wird