RESUMO
BACKGROUND: Per- and polyfluoroalkyl substances (PFAS) are environmental contaminants that have received significant public attention. PFAS are a large group of human-made chemicals that have been used in industry and consumer products worldwide since the 1950s. Human exposure to PFAS is a growing public health concern. Studies suggest that exposure to PFAS may increase the risk of some cancers and have negative health impacts on the endocrine, metabolic, and immune systems. Federal and state health partners are investigating the exposure to and possible health effects associated with PFAS. Government agencies can observe social media discourse on PFAS to better understand public concerns and develop targeted communication and outreach efforts. OBJECTIVE: The primary objective of this study is to understand how social media is used to share, disseminate, and engage in public discussions of PFAS-related information in the United States. METHODS: We investigated PFAS-related content across 2 social media platforms between May 1, 2017, and April 30, 2019, to identify how social media is used in the United States to seek and disseminate PFAS-related information. Our key variable of interest was posts that mentioned "PFAS," "PFOA," "PFOS," and their hashtag variations across social media platforms. Additional variables included post type, time, PFAS event, and geographic location. We examined term use and post type differences across platforms. We used descriptive statistics and regression analysis to assess the incidence of PFAS discussions and to identify the date, event, and geographic patterns. We qualitatively analyzed social media content to determine the most prevalent themes discussed on social media platforms. RESULTS: Our analysis revealed that Twitter had a significantly greater volume of PFAS-related posts compared with Reddit (98,264 vs 3126 posts). PFAS-related social media posts increased by 670% over 2 years, indicating a marked increase in social media users' interest in and awareness of PFAS. Active engagement varied across platforms, with Reddit posts demonstrating more in-depth discussions compared with passive likes and reposts among Twitter users. Spikes in PFAS discussions were evident and connected to the discovery of contamination events, media coverage, and scientific publications. Thematic analysis revealed that social media users see PFAS as a significant public health concern and seek a trusted source of information about PFAS-related public health efforts. CONCLUSIONS: The analysis identified a prevalent theme-on social media, PFAS are perceived as an immediate public health concern, which demonstrates a growing sense of urgency to understand this emerging contaminant and its potential health impacts. Government agencies can continue using social media research to better understand the changing community sentiment on PFAS and disseminate targeted information and then use social media as a forum for dispelling misinformation, communicating scientific findings, and providing resources for relevant public health services.
Assuntos
Mídias Sociais , Comunicação , Humanos , Infodemiologia , Saúde Pública , Opinião Pública , Estados UnidosRESUMO
The N-terminally myristoylated matrix (MA) domain of the HIV-1 Gag polyprotein promotes virus assembly by targeting Gag to the inner leaflet of the plasma membrane. Recent studies indicate that, prior to membrane binding, MA associates with cytoplasmic tRNAs (including tRNALys3), and in vitro studies of tRNA-dependent MA interactions with model membranes have led to proposals that competitive tRNA interactions contribute to membrane discrimination. We have characterized interactions between native, mutant, and unmyristylated (myr-) MA proteins and recombinant tRNALys3 by NMR spectroscopy and isothermal titration calorimetry. NMR experiments confirm that tRNALys3 interacts with a patch of basic residues that are also important for binding to the plasma membrane marker, phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. Unexpectedly, the affinity of MA for tRNALys3 (Kd = 0.63 ± 0.03 µM) is approximately 1 order of magnitude greater than its affinity for PI(4,5)P2-enriched liposomes (Kd(apparent) = 10.2 ± 2.1 µM), and NMR studies indicate that tRNALys3 binding blocks MA association with liposomes, including those enriched with PI(4,5)P2, phosphatidylserine, and cholesterol. However, the affinity of MA for tRNALys3 is diminished by mutations or sample conditions that promote myristate exposure. Since Gag-Gag interactions are known to promote myristate exposure, our findings support virus assembly models in which membrane targeting and genome binding are mechanistically coupled.
Assuntos
HIV-1/fisiologia , Fosfatidilinositol 4,5-Difosfato/metabolismo , RNA de Transferência/metabolismo , Produtos do Gene gag do Vírus da Imunodeficiência Humana/química , Produtos do Gene gag do Vírus da Imunodeficiência Humana/metabolismo , Calorimetria , Membrana Celular/metabolismo , Citoplasma/genética , HIV-1/genética , HIV-1/metabolismo , Humanos , Espectroscopia de Ressonância Magnética , Modelos Moleculares , Mutação , Ácido Mirístico/metabolismo , Domínios Proteicos , RNA de Transferência/química , RNA de Transferência/genética , Montagem de Vírus , Produtos do Gene gag do Vírus da Imunodeficiência Humana/genéticaRESUMO
Assembly of HIV-1 particles is initiated by the trafficking of viral Gag polyproteins from the cytoplasm to the plasma membrane, where they co-localize and bud to form immature particles. Membrane targeting is mediated by the N-terminally myristoylated matrix (MA) domain of Gag and is dependent on the plasma membrane marker phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. Recent studies revealed that PI(4,5)P2 molecules containing truncated acyl chains [tr-PI(4,5)P2] are capable of binding MA in an "extended lipid" conformation and promoting myristoyl exposure. Here we report that tr-PI(4,5)P2 molecules also readily bind to non-membrane proteins, including HIV-1 capsid, which prompted us to re-examine MA-PI(4,5)P2 interactions using native lipids and membrane mimetic liposomes and bicelles. Liposome binding trends observed using a recently developed NMR approach paralleled results of flotation assays, although the affinities measured under the equilibrium conditions of NMR experiments were significantly higher. Native PI(4,5)P2 enhanced MA binding to liposomes designed to mimic non-raft-like regions of the membrane, suggesting the possibility that binding of the protein to disordered domains may precede Gag association with, or nucleation of, rafts. Studies with bicelles revealed a subset of surface and myr-associated MA residues that are sensitive to native PI(4,5)P2, but cleft residues that interact with the 2'-acyl chains of tr-PI(4,5)P2 molecules in aqueous solution were insensitive to native PI(4,5)P2 in bicelles. Our findings call to question extended-lipid MA:membrane binding models, and instead support a model put forward from coarse-grained simulations indicating that binding is mediated predominantly by dynamic, electrostatic interactions between conserved basic residues of MA and multiple PI(4,5)P2 and phosphatidylserine molecules.
Assuntos
HIV-1/fisiologia , Produtos do Gene gag do Vírus da Imunodeficiência Humana/química , Membrana Celular/metabolismo , Lipídeos/química , Lipossomos/química , Espectroscopia de Ressonância Magnética , Microdomínios da Membrana , Fosfatidilinositol 4,5-Difosfato/metabolismo , Fosfatidilserinas/química , Ligação Proteica , Estrutura Terciária de Proteína , Produtos do Gene gag do Vírus da Imunodeficiência Humana/metabolismoRESUMO
The matrix (MA) domain of the human immunodeficiency virus (HIV) 1 Gag is responsible for Gag targeting to the plasma membrane where virions assemble. MA also plays a role in the incorporation of the viral envelope (Env) glycoproteins and can influence particle infectivity post-maturation and post-entry. A highly basic region of MA targets Gag to the plasma membrane via specific binding to phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2]. This binding also triggers exposure of an amino-terminal myristate moiety, which anchors Gag to the membrane. An MA mutant deficient for PI(4,5)P2 binding, 29KE/31KE, has been shown to mislocalize within the cell, leading to particle assembly in a multivesicular body compartment and defective release of cell-free particles in HeLa and 293T cells. Despite the defect in virus production in these cells, release of the 29KE/31KE mutant is not significantly reduced in primary T cells, macrophages and Jurkat T cells. 29KE/31KE virions also display an infectivity defect associated with impaired Env incorporation, irrespective of the producer cell line. Here we examine the properties of 29KE/31KE by analyzing compensatory mutations obtained by a viral adaptation strategy. The MA mutant 16EK restores virus release through enhanced membrane binding. 16EK also influences the infectivity defect, in combination with an additional MA mutant, 62QR. Additionally, the 29KE/31KE MA mutant displays a defect in proteolytic cleavage of the murine leukemia virus Env cytoplasmic tail in pseudotyped virions. Our findings elucidate the mechanism whereby an MA mutant defective in PI(4,5)P2 binding can be rescued and highlight the ability of MA to influence Env glycoprotein function.