Using viral vectors as gene transfer tools (Cell Biology and Toxicology Special Issue: ETCS-UK 1 day meeting on genetic manipulation of cells)

In recent years, the development of powerful viral gene transfer techniques has greatly facilitated the study of gene function. This review summarises some of the viral delivery systems routinely used to mediate gene transfer into cell lines, primary cell cultures and in whole animal models. The systems described were originally discussed at a 1-day European Tissue Culture Society (ETCS-UK) workshop that was held at University College London on 1st April 2009. Recombinant-deficient viral vectors (viruses that are no longer able to replicate) are used to transduce dividing and post-mitotic cells, and they have been optimised to mediate regulatable, powerful, long-term and cell-specific expression. Hence, viral systems have become very widely used, especially in the field of neurobiology. This review introduces the main categories of viral vectors, focusing on their initial development and highlighting modifications and improvements made since their introduction. In particular, the use of specific promoters to restrict expression, translational enhancers and regulatory elements to boost expression from a single virion and the development of regulatable systems is described

Studio del profilo aromatico di oli extra vergini di oliva della cultivar Nostrana di Brisighella

L'olio extra vergine di oliva è apprezzato per i suoi benefici per la salute, le sue proprietà nutrizionali e caratteristiche organolettiche. La sua qualità dipende da diversi fattori, tra cui la varietà delle olive, la zona di coltivazione, le pratiche agronomiche, le tecnologie di trasformazione e le condizioni di conservazione. Questa tesi si propone di analizzare la frazione volatile di dieci oli extra vergini di oliva prodotti presso il Campus di Scienze degli Alimenti di Cesena, utilizzando il frantoio da banco Abencor®. Le olive della cultivar "Nostrana di Brisighella" provengono da diverse aree di uno stesso uliveto a Brisighella. Sono stati utilizzati due metodi gas cromatografici per l’analisi della frazione volatile degli oli in esame: la microestrazione in fase solida accoppiata alla gas cromatografia e spettrometria di massa (SPME-GC-MS) e la flash-gas cromatografia (FGC-E-nose). Queste tecniche hanno permesso di confrontare i profili aromatici degli oli prodotti e identificare eventuali analogie e differenze. Si è prestata particolare attenzione a valutare l’influenza della posizione degli olivi sulla composizione della frazione volatile degli oli, contribuendo ad indagare il legame tra il profilo aromatico degli oli e il territorio di origine. È importante notare che gli oli sono stati prodotti in due giornate consecutive per ragioni logistiche, pertanto è stata valutata l’influenza dei diversi tempi di conservazione delle olive sul profilo in composti volatili degli oli. I risultati ottenuti, che si riferiscono esclusivamente alla composizione della frazione volatile degli oli prodotti, saranno integrati con quelli di altre analisi, attualmente in corso, di qualità degli oli e relative alle caratteristiche compositive del suolo e delle foglie di ulivo. In questo modo sarà possibile stabilire se la posizione degli ulivi in campo o la durata della conservazione delle olive abbiano influenzato la composizione degli oli prodotti

Human Immunodeficiency Virus Type 1 Latency Model for High-Throughput Screening

Human immunodeficiency virus type 1 (HIV-1) is not eliminated from patients even after years of antiretroviral therapy, apparently due to the presence of latently infected cells. Here we describe the development of a cell-based system of latency that can be used for high-throughput screening aimed at novel drug discovery to eradicate HIV-1 infection

Toward the Development of a Virus-Cell-Based Assay for the Discovery of Novel Compounds against Human Immunodeficiency Virus Type 1

The emergence of human immunodeficiency virus type 1 (HIV-1) strains resistant to highly active antiretroviral therapy necessitates continued drug discovery for the treatment of HIV-1 infection. Most current drug discovery strategies focus upon a single aspect of HIV-1 replication. A virus-cell-based assay, which can be adapted to high-throughput screening, would allow the screening of multiple targets simultaneously. HIV-1-based vector systems mimic the HIV-1 life cycle without yielding replication-competent virus, making them potentially important tools for the development of safe, wide-ranging, rapid, and cost-effective assays amenable to high-throughput screening. Since replication of vector virus is typically restricted to a single cycle, a crucial question is whether such an assay provides the needed sensitivity to detect potential HIV-1 inhibitors. With a stable, inducible vector virus-producing cell line, the inhibitory effects of four reverse transcriptase inhibitors (zidovudine, stavudine, lamivudine, and didanosine) and one protease inhibitor (indinavir) were assessed. It was found that HIV-1 vector virus titer was inhibited in a single cycle of replication up to 300-fold without affecting cell viability, indicating that the assay provides the necessary sensitivity for identifying antiviral molecules. Thus, it seems likely that HIV-1-derived vector systems can be utilized in a novel fashion to facilitate the development of a safe, efficient method for screening compound libraries for anti-HIV-1 activity