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1.
ISME J ; 14(3): 727-739, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31822788

RESUMO

Acanthamoeba-infecting Mimiviridae are giant viruses with dsDNA genome up to 1.5 Mb. They build viral factories in the host cytoplasm in which the nuclear-like virus-encoded functions take place. They are themselves the target of infections by 20-kb-dsDNA virophages, replicating in the giant virus factories and can also be found associated with 7-kb-DNA episomes, dubbed transpovirons. Here we isolated a virophage (Zamilon vitis) and two transpovirons respectively associated to B- and C-clade mimiviruses. We found that the virophage could transfer each transpoviron provided the host viruses were devoid of a resident transpoviron (permissive effect). If not, only the resident transpoviron originally isolated from the corresponding virus was replicated and propagated within the virophage progeny (dominance effect). Although B- and C-clade viruses devoid of transpoviron could replicate each transpoviron, they did it with a lower efficiency across clades, suggesting an ongoing process of adaptive co-evolution. We analysed the proteomes of host viruses and virophage particles in search of proteins involved in this adaptation process. This study also highlights a unique example of intricate commensalism in the viral world, where the transpoviron uses the virophage to propagate and where the Zamilon virophage and the transpoviron depend on the giant virus to replicate, without affecting its infectious cycle.


Assuntos
Acanthamoeba/virologia , Mimiviridae/fisiologia , Vírus Gigantes/genética , Vírus Gigantes/fisiologia , Mimiviridae/genética , Mimiviridae/crescimento & desenvolvimento , Mimiviridae/isolamento & purificação , Simbiose , Virófagos/genética , Virófagos/fisiologia
2.
Nat Commun ; 9(1): 2285, 2018 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-29891839

RESUMO

With DNA genomes reaching 2.5 Mb packed in particles of bacterium-like shape and dimension, the first two Acanthamoeba-infecting pandoraviruses remained up to now the most complex viruses since their discovery in 2013. Our isolation of three new strains from distant locations and environments is now used to perform the first comparative genomics analysis of the emerging worldwide-distributed Pandoraviridae family. Thorough annotation of the genomes combining transcriptomic, proteomic, and bioinformatic analyses reveals many non-coding transcripts and significantly reduces the former set of predicted protein-coding genes. Here we show that the pandoraviruses exhibit an open pan-genome, the enormous size of which is not adequately explained by gene duplications or horizontal transfers. As most of the strain-specific genes have no extant homolog and exhibit statistical features comparable to intergenic regions, we suggest that de novo gene creation could contribute to the evolution of the giant pandoravirus genomes.


Assuntos
Acanthamoeba/virologia , Vírus de DNA/classificação , Vírus de DNA/genética , Vírus de DNA/fisiologia , DNA Viral/genética , Microbiologia Ambiental , Evolução Molecular , Duplicação Gênica , Transferência Genética Horizontal , Variação Genética , Genoma Viral , Anotação de Sequência Molecular , Filogenia , Proteômica , Análise de Sequência de DNA , Vírion/ultraestrutura , Replicação Viral
3.
Drug Test Anal ; 9(2): 179-187, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26990774

RESUMO

Currently under development for therapeutic purposes in human medicine, non-steroidal selective androgen receptor modulators (non-steroidal SARMs) are also known to impact growth associated pathways. As such, they present a potential for abuse in sports and food-producing animals as interesting alternative anabolic substances. Forbidden since 2008 by the World Anti-Doping Agency (WADA) these compounds are however easily available and could be (mis)used in livestock production as growth promoters. To prevent such practices, dedicated analytical strategies have to be developed for specific and sensitive detection of these compounds in biological matrices. Using an innovative analytical platform constituted of supercritical fluid chromatography coupled to ion mobility-mass spectrometry, the present study enabled efficient separation and identification in urine of 4 of these drugs (andarine, bicalutamide, hydroxyflutamide, and enobosarm) in accordance with European Union criteria (Commission Decision 2002/657/EC). Besides providing information about compounds structure and behaviour in gas phase, such a coupling enabled reaching low limits of detection (LOD < 0.05 ng.mL-1 for andarine and limits of detection < 0.005 ng.mL-1 for the three others) in urine with good repeatability (CV < 21 %). The workflow has been applied to quantitative determination of enobosarm elimination in urine of treated bovine (200 mg, oral). Copyright © 2016 John Wiley & Sons, Ltd.


Assuntos
Anabolizantes/urina , Androgênios/urina , Anilidas/urina , Bovinos/urina , Cromatografia com Fluido Supercrítico/métodos , Espectrometria de Massas/métodos , Detecção do Abuso de Substâncias/métodos , Animais , Dopagem Esportivo , Limite de Detecção , Masculino
4.
Anal Chem ; 87(18): 9234-42, 2015 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-26305505

RESUMO

While the coupling of traveling wave ion mobility spectrometry (TWIMS) and mass spectrometry is mainly reported for structural purposes, we studied its potential in enhancing compounds analysis such as growth promoters used in livestock animals at trace concentrations. ß-Adrenergic agonists have been selected as model compounds since they exhibit a range of close physicochemical properties leading to analytical issues using classical approaches. In this paper, the potential of Synapt G2-S (Q-TWIM-TOF MS) has been investigated for sensitive and specific detection of a range of these synthetic phenethanolamines in various complex biological matrices (retina, meat, and urine) from bovine considered as relevant in the context of detecting ß-adrenergic agonists use in animals. In particular, the specificity of the additional information provided by the TWIMS (i.e., collision cross section) together with the interest of the extra dimension of separation is discussed.


Assuntos
Agonistas Adrenérgicos beta/análise , Agonistas Adrenérgicos beta/isolamento & purificação , Espectrometria de Massas/métodos , Agonistas Adrenérgicos beta/química , Agonistas Adrenérgicos beta/urina , Animais , Bovinos , Bases de Dados de Produtos Farmacêuticos , Limite de Detecção , Peso Molecular , Carne Vermelha/análise , Retina/química , Fatores de Tempo
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