HIV-1 proviral landscape characterization varies by pipeline analysis.

INTRODUCTION: HIV rebounds after cessation of antiretroviral therapy, representing a barrier to cure. To better understand the virus reservoir, analysis pipelines have been developed that categorize proviral sequences as intact or defective, and further determine the precise nature of the sequence defects that may be present. We investigated the effects that different analysis pipelines had on the characterization of HIV-1 proviral sequences. METHODS: We used single genome amplification to generate near full-length (NFL) HIV-1 proviral DNA sequences, defined as amplicons greater than 8000 base pairs in length, isolated from peripheral blood mononuclear cells (PBMC) of treated suppressed participants with HIV-1. Amplicons underwent direct next-generation single genome sequencing and were analysed using four HIV-1 proviral characterization pipelines. Sequences were characterized as intact or defective; defective sequences were assessed for the number and types of defects present. To confirm and extend our findings, 691 proviruses from the Proviral Sequence Database (PSD) were analysed and the ProSeq-IT tool of the PSD was used to characterize both the participant and PSD proviruses. RESULTS AND DISCUSSION: Virus sequences derived from thirteen ART-treated virologically suppressed participants with HIV were studied. A total of 693 HIV-1 proviral sequences were generated, 282 of which were NFL. An average of 53 sequences per participant was analysed. We found that proviruses often harbour multiple sequence defect types (mean 2.7, 95% confidence interval [CI] 2.5, 3.0); the elimination order used by each pipeline affected the percentage of proviruses allotted into each defect category. These differences varied between participants, depending on the number of defect categories present in a given provirus sequence. Pipeline-specific differences in characterizing the HIV-1 5' untranslated region (5' UTR) led to an overestimation of the number of intact NFL proviral sequences, a finding corroborated in the independent PSD analysis. A comparison of the four published pipelines to ProSeq-IT found that ProSeq IT was more likely to characterize proviruses as intact. CONCLUSIONS: The choice of pipeline used for HIV-1 provirus landscape analysis may bias the classification of defective sequences. To improve the comparison of provirus characterizations across research groups, the development of a consensus elimination pipeline should be prioritized.

Arsenic biotransformation by cyanobacteria from mining areas: evidences from culture experiments.

Elucidating the role of cyanobacteria in the biotransformation of arsenic (As) oxyanions is crucial to understand the biogeochemical cycle of this element and indicate species with potential for its bioremediation. In this study, we determined the EC50 for As(III) and As(V) and evaluated the biotransformation of As by Synechococcus sp. through high-performance liquid chromatography hyphenated to inductively coupled plasma mass spectrometry (HPLC-ICP-MS) and X-ray absorption fine structure spectroscopy (XAFS). Synechococcus sp. exhibited higher sensitivity to As(III) with an EC(50, 96 h) of 6.64 mg L(-1) that was approximately 400-fold lower than that for As(V). Even though the cells were exposed to concentrations of As(III) (6 mg L(-1)) approximately 67-fold lower than those of As(V) (400 mg L(-1)), similar intracellular concentrations of As (60.0 µg g(-1)) were observed after 30 days. As(V) was the predominant intracellular As species followed by As(III). Furthermore, organic As species such as monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA) were observed in higher proportions after exposure to As(III). The differential toxicity among As oxyanions indicates that determining the redox state of As in the environment is fundamental to estimate toxicity risks to aquatic organisms. Synechococcus sp. demonstrated potential for its application in bioremediation due to the high accumulation of As and production of As organic compounds notably after exposure to As(III).

Characterization of a novel tymovirus on tomato plants in Brazil.

A tymovirus was isolated in Brazil from tomato plants with severe symptoms of leaf mosaic and blistering. The virus was mechanically transmissible to solanaceous indicator host species. The infected plants contained icosahedral particles and chloroplasts with membrane deformations which are typical cytopathic effects caused by tymoviruses. Its coat protein amino acid sequence shares the maximum of 64 % identity with the tymovirus Chiltepin yellow mosaic virus, which suggested that it can be considered as a distinct member of the genus Tymovirus. In a phylogenetic tree, this tymovirus was clustered with other solanaceous-infecting tymoviruses. It was tentatively named as Tomato blistering mosaic virus (ToBMV).