RESUMO
This review summarizes our current knowledge on the health and environmental impact as well as the mineralogical and geochemical composition of nanoparticles (NPs) associated with coal fires. It will furthermore recommend new sampling and characterization protocols to gain a better understanding of the various types of NPs that are formed either through high-temperature nucleation and alteration processes or via low-temperature dissolution-reprecipitation and weathering processes. Coal fires affect the immediate environment of coal-producing areas and produce positive and negative feedback to climate change through the emission of carbon- and sulfate-bearing gases and aerosols, respectively. Nanoparticles form during and after coal fires. They are composed of mainly soot and tar particles as well as amorphous phases, minerals, and complex mixtures of amorphous phases and minerals. It is recommended that NPs for mineralogical studies should be collected using impactors (a new generation of collectors for particulate matter, such as the TPS100 NP sampler) or that borosilicate filters at the openings of pipes and chambers be used to collect and measure gases emitted by coal fires. Furthermore, assemblages of NPs occurring at the mouths of coal fire vents should be examined using a combination of focused ion beam (FIB) technology and transmission electron microscopy (TEM), and those containing ion- or electron-beam sensitive phases should be examined with the corresponding cryo-techniques, such as cryo-FIB, cryo-ion mill, and cryo-TEM. The mineralogical and chemical composition of NP-bearing bulk samples should be examined with spectroscopy techniques such as X-ray photoelectron spectroscopy, 13 C nuclear magnetic resonance spectroscopy, or time-of-flight secondary ion mass spectroscopy.
Assuntos
Incêndios , Nanopartículas , Carvão Mineral , Carbono , GasesRESUMO
Peribunyaviridae is a large family of RNA viruses with several members that cause mild to severe diseases in humans and livestock. Despite their importance in public heath very little is known about the host cell factors hijacked by these viruses to support assembly and cell egress. Here we show that assembly of Oropouche virus, a member of the genus Orthobunyavirus that causes a frequent arboviral infection in South America countries, involves budding of virus particles toward the lumen of Golgi cisternae. As viral replication progresses, these Golgi subcompartments become enlarged and physically separated from Golgi stacks, forming Oropouche viral factory (Vfs) units. At the ultrastructural level, these virally modified Golgi cisternae acquire an MVB appearance, and while they lack typical early and late endosome markers, they become enriched in endosomal complex required for transport (ESCRT) proteins that are involved in MVB biogenesis. Further microscopy and viral replication analysis showed that functional ESCRT machinery is required for efficient Vf morphogenesis and production of infectious OROV particles. Taken together, our results indicate that OROV attracts ESCRT machinery components to Golgi cisternae to mediate membrane remodeling events required for viral assembly and budding at these compartments. This represents an unprecedented mechanism of how viruses hijack host cell components for coordinated morphogenesis.
Assuntos
Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Orthobunyavirus/metabolismo , Orthobunyavirus/fisiologia , Técnicas de Cultura de Células , Complexos Endossomais de Distribuição Requeridos para Transporte/fisiologia , Endossomos/metabolismo , Complexo de Golgi/metabolismo , Complexo de Golgi/virologia , Células HeLa , Humanos , Orthobunyavirus/crescimento & desenvolvimento , Orthobunyavirus/patogenicidade , Vírion/metabolismo , Montagem de Vírus/fisiologia , Liberação de Vírus/fisiologia , Replicação Viral/fisiologiaRESUMO
HIV-1 Nef-mediated CD4 downmodulation involves various host factors. We investigated the importance of AP-1, AP-2, AP-3, V1H-ATPase, ß-COP, and ACOT8 for CD4 downmodulation in HIV-1-infected short hairpin RNA (shRNA)-expressing CD4(+) T cells and characterized direct interaction with Nef by Förster resonance energy transfer (FRET). Binding of lentiviral Nefs to CD4 and AP-2 was conserved, and only AP-2 knockdown impaired Nef-mediated CD4 downmodulation from primary T cells. Altogether, among the factors tested, AP-2 is the most important player for Nef-mediated CD4 downmodulation.
Assuntos
Complexo 2 de Proteínas Adaptadoras/metabolismo , Antígenos CD4/biossíntese , Linfócitos T CD4-Positivos/metabolismo , Regulação para Baixo , Infecções por HIV/metabolismo , HIV-1/metabolismo , Produtos do Gene nef do Vírus da Imunodeficiência Humana/metabolismo , Complexo 2 de Proteínas Adaptadoras/genética , Complexo 2 de Proteínas Adaptadoras/imunologia , Antígenos CD4/genética , Antígenos CD4/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Técnicas de Silenciamento de Genes , Infecções por HIV/genética , Infecções por HIV/imunologia , Infecções por HIV/patologia , HIV-1/genética , HIV-1/imunologia , Humanos , Produtos do Gene nef do Vírus da Imunodeficiência Humana/genética , Produtos do Gene nef do Vírus da Imunodeficiência Humana/imunologiaRESUMO
HIV Nef-mediated up-regulation of invariant chain (Ii chain, also CD74) is presumed to play an active role in HIV immunopathogenesis. However, this has not been definitely ascertained. In order to help elucidate this hypothesis, Ii chain, CD4, HLA-DR and HLA-ABC expression was analyzed ex vivo in monocyte-derived macrophages (MDMs) from HIV(+) subjects. Viral load, CD4(+) T cell count and immune activation were also determined in enrolled subjects. Correlations between these parameters and the modulation of cell surface molecules in infected cells were studied. Ii chain expression was found to be up-regulated in infected MDMs derived from all patients but one (median fold up-regulation 2.47±1.82 (range 0.87-7.36)). Moreover, the magnitude of Ii chain up-regulation significantly correlated with higher activation of B and CD4(+) T cells (studied by HLA-DR and CD38 expression). On the other hand, lower HLA-ABC (i.e. stronger down-regulation) in infected MDMs was associated with higher CD4 counts. No correlation was observed between the magnitude of Ii chain up-regulation and the other Nef functions studied here. This is the first study reporting that Ii chain up-regulation occurs on naturally infected antigen presenting cells obtained directly from HIV(+) subjects. Moreover, it is also shown that the magnitude of this up-regulation correlates with immune activation. This allows postulating an alternative hypothesis regarding the contribution of Ii chain up-regulation to HIV-mediated immune damage.
Assuntos
Síndrome da Imunodeficiência Adquirida/imunologia , Síndrome da Imunodeficiência Adquirida/patologia , Antígenos de Diferenciação de Linfócitos B/biossíntese , HIV/imunologia , HIV/patogenicidade , Antígenos de Histocompatibilidade Classe II/biossíntese , ADP-Ribosil Ciclase 1/análise , Síndrome da Imunodeficiência Adquirida/virologia , Linfócitos B/química , Linfócitos B/imunologia , Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos/química , Linfócitos T CD4-Positivos/imunologia , Células Cultivadas , Perfilação da Expressão Gênica , Antígenos HLA-DR/análise , Humanos , Macrófagos/imunologia , Macrófagos/virologia , Glicoproteínas de Membrana/análise , Regulação para Cima , Carga ViralRESUMO
HIV-1 Nef protein plays a major role in viral immunopathogenesis, modulating surface expression of several immune receptors, altering signal transduction pathways, and enhancing viral infectivity, among other activities. Nef also exhibits great intersubtype diversity, but most studies have been focused only on Nef proteins from subtype B. Thus, little is known about the functional capacities of nonsubtype B Nef proteins in host cells. Here, we investigated cell surface regulation of MHC-I, MHC-II, the MHC-II-associated chaperone invariant chain (Ii), CD4, CD3, and CD28 in cells transfected or infected with five different Nef alleles including one HIV-1 subtype C and F allele. No significant difference among the Nef proteins regarding CD3, CD28, and MHC-II downregulation was observed. The NefC showed a slightly, yet significant, diminished capacity to downregulate MHC-I in all cells, as well as to downregulate CD4 in Jurkat cells and PBMCs. Strikingly, the two alleles from NefC and NefF were unable to upregulate the Ii chain both in transfected and infected cells. Moreover, the internalization rate of the surface Ii chain was only slightly affected by NefC and NefF, whereas it was drastically reduced by NefB. Nef domains known to be involved in Ii chain upregulation were conserved among the five alleles analyzed here. In summary, we identified two primary HIV-1 NefC and NefF alleles that are selectively impaired for Ii upregulation and that may help to elucidate the mechanism of this Nef function in the future. It will be important to determine whether the observed differences are HIV-1 subtype dependent and influence viral immunopathogenesis.