Browsing by Subject "Reverse transcriptase-polymerase chain reaction"

Now showing 1 - 4 of 4
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    Open Access
    Accumulation of splice variants and transcripts in response to PI3K inhibition in T cells
    (Public Library of Science, 2013) Riedel, Alice; Mofolo, Boitumelo; Avota, Elita; Schneider-Schaulies, Sibylle; Meintjes, Ayton; Mulder, Nicola; Kneitz, Susanne
    BACKGROUND: Measles virus (MV) causes T cell suppression by interference with phosphatidylinositol-3-kinase (PI3K) activation. We previously found that this interference affected the activity of splice regulatory proteins and a T cell inhibitory protein isoform was produced from an alternatively spliced pre-mRNA. Hypothesis Differentially regulated and alternatively splice variant transcripts accumulating in response to PI3K abrogation in T cells potentially encode proteins involved in T cell silencing. METHODS: To test this hypothesis at the cellular level, we performed a Human Exon 1.0 ST Array on RNAs isolated from T cells stimulated only or stimulated after PI3K inhibition. We developed a simple algorithm based on a splicing index to detect genes that undergo alternative splicing (AS) or are differentially regulated (RG) upon T cell suppression. RESULTS: Applying our algorithm to the data, 9% of the genes were assigned as AS, while only 3% were attributed to RG. Though there are overlaps, AS and RG genes differed with regard to functional regulation, and were found to be enriched in different functional groups. AS genes targeted extracellular matrix (ECM)-receptor interaction and focal adhesion pathways, while RG genes were mainly enriched in cytokine-receptor interaction and Jak-STAT. When combined, AS/RG dependent alterations targeted pathways essential for T cell receptor signaling, cytoskeletal dynamics and cell cycle entry. CONCLUSIONS: PI3K abrogation interferes with key T cell activation processes through both differential expression and alternative splicing, which together actively contribute to T cell suppression.
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    Assessment of environmental contamination and environmental decontamination practices within an Ebola holding unit, Freetown, Sierra Leone
    (Public Library of Science, 2015) Youkee, Daniel; Brown, Colin S; Lilburn, Paul; Shetty, Nandini; Brooks, Tim; Simpson, Andrew; Bentley, Neil; Lado, Marta; Kamara, Thaim B; Walker, Naomi F; Johnson, Oliver
    Evidence to inform decontamination practices at Ebola holding units (EHUs) and treatment centres is lacking. We conducted an audit of decontamination procedures inside Connaught Hospital EHU in Freetown, Sierra Leone, by assessing environmental swab specimens for evidence of contamination with Ebola virus by RT-PCR. Swabs were collected following discharge of Ebola Virus Disease (EVD) patients before and after routine decontamination. Prior to decontamination, Ebola virus RNA was detected within a limited area at all bedside sites tested, but not at any sites distant to the bedside. Following decontamination, few areas contained detectable Ebola virus RNA. In areas beneath the bed there was evidence of transfer of Ebola virus material during cleaning. Retraining of cleaning staff reduced evidence of environmental contamination after decontamination. Current decontamination procedures appear to be effective in eradicating persistence of viral RNA. This study supports the use of viral swabs to assess Ebola viral contamination within the clinical setting. We recommend that regular refresher training of cleaning staff and audit of environmental contamination become standard practice at all Ebola care facilities during EVD outbreaks.
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    Metagenomic-based screening and molecular characterization of cowpea-infecting viruses in Burkina Faso
    (Public Library of Science, 2016) Palanga, Essowè; Filloux, Denis; Martin, Darren P; Fernandez, Emmanuel; Gargani, Daniel; Ferdinand, Romain; Zabré, Jean; Bouda, Zakaria; Neya, James Bouma; Sawadogo, Mahamadou; Traore, Oumar; Peterschmitt, Michel; Roumagnac, Philippe
    Cowpea, ( Vigna unguiculata L. (Walp)) is an annual tropical grain legume. Often referred to as "poor man's meat", cowpea is one of the most important subsistence legumes cultivated in West Africa due to the high protein content of its seeds. However, African cowpea production can be seriously constrained by viral diseases that reduce yields. While twelve cowpea-infecting viruses have been reported from Africa, only three of these have so-far been reported from Burkina Faso. Here we use a virion-associated nucleic acids (VANA)-based metagenomics method to screen for the presence of cowpea viruses from plants collected from the three agro-climatic zones of Burkina Faso. Besides the three cowpea-infecting virus species which have previously been reported from Burkina Faso (Cowpea aphid borne mosaic virus [Family Potyviridae ], the Blackeye cowpea mosaic virus--a strain of Bean common mosaic virus--[Family Potyviridae ] and Cowpea mottle virus [Family Tombusviridae ]) five additional viruses were identified: Southern cowpea mosaic virus (Sobemovirus genus), two previously uncharacterised polerovirus-like species (Family Luteoviridae ), a previously uncharacterised tombusvirus-like species (Family Tombusviridae ) and a previously uncharacterised mycotymovirus-like species (Family Tymoviridae ). Overall, potyviruses were the most prevalent cowpea viruses (detected in 65.5% of samples) and the Southern Sudan zone of Burkina Faso was found to harbour the greatest degrees of viral diversity and viral prevalence. Partial genome sequences of the two novel polerovirus-like and tombusvirus-like species were determined and RT-PCR primers were designed for use in Burkina Faso to routinely detect all of these cowpea-associated viruses.
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    A novel diagnostic target in the hepatitis C virus genome
    (Public Library of Science, 2009) Drexler, Jan Felix; Kupfer, Bernd; Petersen, Nadine; Grotto, Rejane Maria Tommasini; Rodrigues, Silvia Maria Corvino; Grywna, Klaus; Panning, Marcus; Annan, Augustina; Silva, Giovanni Faria; Douglas, Jill
    Christian Drosten and colleagues develop, validate, and make openly available a prototype hepatitis C virus assay based on the conserved 3' X-tail element, with potential for clinical use in developing countries.