380 research outputs found
NETWORKED2âSubfamily Proteins Regulate the Cortical Actin Cytoskeleton of Growing Pollen Tubes and Polarised Pollen Tube Growth
We have recently characterised NET2A as a pollenâspecific actinâbinding protein which binds Fâactin at the plasma membrane of growing pollen tubes. However, the role of NET2 proteins in pollen development and fertilisation have yet to be elucidated. To further characterise the role of Arabidopsis NET2 proteins in pollen development and fertilisation, we analysed the subcellular localisation of NET2A over the course of pollen grain development, and investigated the role of the NET2 family using net2 lossâofâfunction mutants. We observed NET2A to localise to the Fâactin cytoskeleton in developing pollen grains as it underwent striking structural reorganisations at specific stages of development and during germination, and pollen tube growth. Furthermore, net2 lossâofâfunction mutants exhibited striking morphological defects in the early stages of pollen tube growth, arising from frequent alterations to pollen tube growth trajectory. We observed defects in the cortical actin cytoskeleton and actinâdriven subcellular processes in net2 mutant pollen tubes. We demonstrate that NET2 proteins are essential for normal actinâdriven pollen development highlighting an important role for the NET2 family members in regulating pollen tube growth during fertilisation
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Effector memory differentiation increases detection of replication-competent HIV-l in resting CD4+ T cells from virally suppressed individuals.
Studies have demonstrated that intensive ART alone is not capable of eradicating HIV-1, as the virus rebounds within a few weeks upon treatment interruption. Viral rebound may be induced from several cellular subsets; however, the majority of proviral DNA has been found in antigen experienced resting CD4+ T cells. To achieve a cure for HIV-1, eradication strategies depend upon both understanding mechanisms that drive HIV-1 persistence as well as sensitive assays to measure the frequency of infected cells after therapeutic interventions. Assays such as the quantitative viral outgrowth assay (QVOA) measure HIV-1 persistence during ART by ex vivo activation of resting CD4+ T cells to induce latency reversal; however, recent studies have shown that only a fraction of replication-competent viruses are inducible by primary mitogen stimulation. Previous studies have shown a correlation between the acquisition of effector memory phenotype and HIV-1 latency reversal in quiescent CD4+ T cell subsets that harbor the reservoir. Here, we apply our mechanistic understanding that differentiation into effector memory CD4+ T cells more effectively promotes HIV-1 latency reversal to significantly improve proviral measurements in the QVOA, termed differentiation QVOA (dQVOA), which reveals a significantly higher frequency of the inducible HIV-1 replication-competent reservoir in resting CD4+ T cells
Interactive molecular dynamics in virtual reality for accurate flexible protein-ligand docking
Simulating drug binding and unbinding is a challenge, as the rugged energy
landscapes that separate bound and unbound states require extensive sampling
that consumes significant computational resources. Here, we describe the use of
interactive molecular dynamics in virtual reality (iMD-VR) as an accurate
low-cost strategy for flexible protein-ligand docking. We outline an
experimental protocol which enables expert iMD-VR users to guide ligands into
and out of the binding pockets of trypsin, neuraminidase, and HIV-1 protease,
and recreate their respective crystallographic protein-ligand binding poses
within 5 - 10 minutes. Following a brief training phase, our studies shown that
iMD-VR novices were able to generate unbinding and rebinding pathways on
similar timescales as iMD-VR experts, with the majority able to recover binding
poses within 2.15 Angstrom RMSD of the crystallographic binding pose. These
results indicate that iMD-VR affords sufficient control for users to carry out
the detailed atomic manipulations required to dock flexible ligands into
dynamic enzyme active sites and recover crystallographic poses, offering an
interesting new approach for simulating drug docking and generating binding
hypotheses.Comment: PLOS ON
Human Galectin-9 Is a Potent Mediator of HIV Transcription and Reactivation.
Identifying host immune determinants governing HIV transcription, latency and infectivity in vivo is critical to developing an HIV cure. Based on our recent finding that the host factor p21 regulates HIV transcription during antiretroviral therapy (ART), and published data demonstrating that the human carbohydrate-binding immunomodulatory protein galectin-9 regulates p21, we hypothesized that galectin-9 modulates HIV transcription. We report that the administration of a recombinant, stable form of galectin-9 (rGal-9) potently reverses HIV latency in vitro in the J-Lat HIV latency model. Furthermore, rGal-9 reverses HIV latency ex vivo in primary CD4+ T cells from HIV-infected, ART-suppressed individuals (p = 0.002), more potently than vorinostat (p = 0.02). rGal-9 co-administration with the latency reversal agent "JQ1", a bromodomain inhibitor, exhibits synergistic activity (p<0.05). rGal-9 signals through N-linked oligosaccharides and O-linked hexasaccharides on the T cell surface, modulating the gene expression levels of key transcription initiation, promoter proximal-pausing, and chromatin remodeling factors that regulate HIV latency. Beyond latent viral reactivation, rGal-9 induces robust expression of the host antiviral deaminase APOBEC3G in vitro and ex vivo (FDR<0.006) and significantly reduces infectivity of progeny virus, decreasing the probability that the HIV reservoir will be replenished when latency is reversed therapeutically. Lastly, endogenous levels of soluble galectin-9 in the plasma of 72 HIV-infected ART-suppressed individuals were associated with levels of HIV RNA in CD4+ T cells (p<0.02) and with the quantity and binding avidity of circulating anti-HIV antibodies (p<0.009), suggesting a role of galectin-9 in regulating HIV transcription and viral production in vivo during therapy. Our data suggest that galectin-9 and the host glycosylation machinery should be explored as foundations for novel HIV cure strategies
The effectiveness of web-based interventions designed to decrease alcohol consumption â a systematic review
OBJECTIVE
To review the published literature on the effectiveness of web-based interventions designed to decrease consumption of alcohol and/or prevent alcohol abuse.
METHOD
Relevant articles published up to, and including, May 2006 were identified through electronic searches of Medline, PsycInfo, Embase, Cochrane Library, ASSIA, Web of Science and Science Direct. Reference lists of all articles identified for inclusion were checked for articles of relevance. An article was included if its stated or implied purpose was to evaluate a web-based intervention designed to decrease consumption of alcohol and/or to prevent alcohol abuse. Studies were reliably selected and quality-assessed, and data were independently extracted and interpreted by two authors.
RESULTS
Initial searches identified 191 articles of which 10 were eligible for inclusion. Of these, five provided a process evaluation only, with the remaining five providing some pre-to post-intervention measure of effectiveness. In general the percentage quality criteria met was relatively low and only one of the 10 articles selected was a randomized control trial.
CONCLUSION
The current review provides inconsistent evidence on the effectiveness of eIectronic screening and brief intervention (eSBI) for alcohol use. Process research suggests that web-based interventions are generally well received. However further controlled trials are needed to fully investigate their efficacy, to determine which elements are keys to outcome and to understand if different elements are required in order to engage low- and high-risk drinkers
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CCR5AS lncRNA variation differentially regulates CCR5, influencing HIV disease outcome.
Multiple genome-wide studies have identified associations between outcome of human immunodeficiency virus (HIV) infection and polymorphisms in and around the gene encoding the HIV co-receptor CCR5, but the functional basis for the strongest of these associations, rs1015164A/G, is unknown. We found that rs1015164 marks variation in an activating transcription factor 1 binding site that controls expression of the antisense long noncoding RNA (lncRNA) CCR5AS. Knockdown or enhancement of CCR5AS expression resulted in a corresponding change in CCR5 expression on CD4+ T cells. CCR5AS interfered with interactions between the RNA-binding protein Raly and the CCR5 3' untranslated region, protecting CCR5 messenger RNA from Raly-mediated degradation. Reduction in CCR5 expression through inhibition of CCR5AS diminished infection of CD4+ T cells with CCR5-tropic HIV in vitro. These data represent a rare determination of the functional importance of a genome-wide disease association where expression of a lncRNA affects HIV infection and disease progression
Actin-membrane interactions mediated by NETWORKED2 in Arabidopsis pollen tubes through associations with Pollen Receptor-Like Kinase 4 and 5
Interactive molecular dynamics in virtual reality from quantum chemistry to drug binding: An open-source multi-person framework
© 2019 Author(s). As molecular scientists have made progress in their ability to engineer nanoscale molecular structure, we face new challenges in our ability to engineer molecular dynamics (MD) and flexibility. Dynamics at the molecular scale differs from the familiar mechanics of everyday objects because it involves a complicated, highly correlated, and three-dimensional many-body dynamical choreography which is often nonintuitive even for highly trained researchers. We recently described how interactive molecular dynamics in virtual reality (iMD-VR) can help to meet this challenge, enabling researchers to manipulate real-time MD simulations of flexible structures in 3D. In this article, we outline various efforts to extend immersive technologies to the molecular sciences, and we introduce "Narupa," a flexible, open-source, multiperson iMD-VR software framework which enables groups of researchers to simultaneously cohabit real-time simulation environments to interactively visualize and manipulate the dynamics of molecular structures with atomic-level precision. We outline several application domains where iMD-VR is facilitating research, communication, and creative approaches within the molecular sciences, including training machines to learn potential energy functions, biomolecular conformational sampling, protein-ligand binding, reaction discovery using "on-the-fly" quantum chemistry, and transport dynamics in materials. We touch on iMD-VR's various cognitive and perceptual affordances and outline how these provide research insight for molecular systems. By synergistically combining human spatial reasoning and design insight with computational automation, technologies such as iMD-VR have the potential to improve our ability to understand, engineer, and communicate microscopic dynamical behavior, offering the potential to usher in a new paradigm for engineering molecules and nano-architectures
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