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The environmental impact of dissolved trace element (DTE) pollution is becoming increasingly well understood. The southeastern edge of the Qinghai-Tibet Plateau is a region with high environmental vulnerability, making it highly susceptible to various anthropogenic disturbances. The Lancang River (LCR), as the most representative river in this area, serving as the largest international river in Southeast Asia, the LCR is crucial for the downstream regions, supporting fisheries, agriculture, and even economic development, earning it the title of the "water tower" for these areas. This study mainly focuses on the upstream unbuilt dam area. In this study, we gathered 25 surface water samples from the upstream of the LCR and examined the geographical distribution, primary sources, and associated human health risks of seven dissolved trace elements (Mn, Cr, Fe, Co, Cu, Ni, Zn). Cr, Fe, Co, Ni, and Cu in the main stream are primarily associated with natural sources, including rock weathering, soil erosion, and similar processes. Mn, however, predominantly originates from human activities such as industrial construction and road transportation. Zn mainly originates from the specialized fertilizers used for highland barley in plateau regions. Almost all DTE concentrations exhibited a downward trend from upstream to downstream. However, due to inputs from various sources along the way, there will be some abnormal points. The concentrations of DTE in the study area were within the acceptable limits set by drinking water standards in China, the USA, and the WHO. Overall, the water quality in this region is considered good. The results from a Monte Carlo simulation assessing health risks indicate that the non-carcinogenic health risk caused by DTE in the upper reaches of Lancang River was very weak. The primary pathway through which DTE can harm human health is ingestion, with children being particularly vulnerable. These findings offer a critical scientific basis for understanding the migration of DTE in aquatic environments and for the ecological stewardship of the Lancang River.
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Hepatitis C virus (HCV) infection remains a challenge to human public health despite the development of highly effective direct-acting antivirals (DAAs). Sofosbuvir (SOF), a key component in most DAA-based anti-HCV cocktail regimens, is a potent viral RNA polymerase (NS5B) inhibitor with a high barrier to drug resistance. The serine-to-threonine mutation at NS5B 282 (S282T) confers the SOF resistance, but severely impairs viral replication in most HCV genotypes (GTs) and cannot be stably maintained after the termination of the SOF-based therapies. In this study, we first developed a new HCV GT-6a subgenomic replicon PR58D6. Next, we selected SOF-resistant PR58D6 variants by culturing the replicon cells in the presence of SOF. Interestingly, unlike many other HCV replicons which require additional mutations to compensate for the S282T-inducing fitness loss, S282T alone in PR58D6 is genetically stable and confers the SOF resistance without significantly impairing viral replication. Furthermore, we showed that amino acid residue at NS5B 74 (R74) and 556 (D556) which are conserved in GT 6a HCV contribute to efficient replication of PR58D6 containing S282T. Finally, we showed that the G556D mutation in NS5B could rescue the replication deficiency of the S282T in JFH1, a GT-2a replicon. In conclusion, we showed that a novel GT-6a HCV replicon may easily render SOF resistance, which may call for attention to potential drug resistance during DAA therapies of HCV GT-6a patients.
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Hepatite C Crônica , Hepatite C , Humanos , Sofosbuvir/farmacologia , RNA Subgenômico , Hepacivirus/genética , Antivirais/farmacologia , Hepatite C/tratamento farmacológico , GenótipoRESUMO
Infection with Ebola virus (EBOV) is responsible for hemorrhagic fever in humans with a high mortality rate. Combined efforts of prevention and therapeutic intervention are required to tackle highly variable RNA viruses, whose infections often lead to outbreaks. Here, we have screened the 2P2I3D chemical library using a nanoluciferase-based protein complementation assay (NPCA) and isolated two compounds that disrupt the interaction of the EBOV protein fragment VP35IID with the N-terminus of the dsRNA-binding proteins PKR and PACT, involved in IFN response and/or intrinsic immunity, respectively. The two compounds inhibited EBOV infection in cell culture as well as infection by measles virus (MV) independently of IFN induction. Consequently, we propose that the compounds are antiviral by restoring intrinsic immunity driven by PACT. Given that PACT is highly conserved across mammals, our data support further testing of the compounds in other species, as well as against other negative-sense RNA viruses.
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Ebolavirus , Doença pelo Vírus Ebola , Humanos , Animais , Doença pelo Vírus Ebola/tratamento farmacológico , Doença pelo Vírus Ebola/metabolismo , Ebolavirus/fisiologia , Antivirais/farmacologia , Antivirais/uso terapêutico , MamíferosRESUMO
Streptococcus sinensis is a recently identified member of the Mitis group of streptococci. This species has been associated with infective endocarditis; however its mechanisms of pathogenesis and virulence are not fully understood. This study aimed to investigate the influence of the competence-stimulating peptide (CSP) and the competence regulon quorum-sensing circuitry (ComABCDE) on subsequent gene transcription and expression, as well as resultant phenotypes. In this study we confirmed the native CSP identity, ascertained when endogenous CSP was produced and completed a transcriptome-wide analysis of all genes following CSP exposure. RNA sequencing analysis revealed the upregulation of genes known to be associated with competence, biofilm formation and virulence. As such, a variety of phenotypic assays were utilized to assess the correlation between increased mRNA expression and potential phenotype response, ultimately gaining insight into the effects of CSP on both gene expression and developed phenotypes. The results indicated that the addition of exogenous CSP aided in competence development and successful transformation, yielding an average transformation efficiency comparable to that of other Mitis group streptococci. Additional studies are needed to further delineate the effects of CSP exposure on biofilm formation and virulence. Overall, this study provides novel information regarding S. sinensis and provides a substantial foundation on which this species and its role in disease pathogenesis can be further investigated.
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Proteínas de Bactérias , Regulon , Proteínas de Bactérias/metabolismo , Percepção de Quorum/genética , Perfilação da Expressão Gênica , Fenótipo , RNA Mensageiro , Regulação Bacteriana da Expressão GênicaRESUMO
Hepatitis C virus (HCV) infection is a major cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. HCV can be sensed by host innate immunity to induce expression of interferons (IFNs) and a number of antiviral effectors. In this study, we found HCV infection induced the expression of neuralized E3 ubiquitin protein ligase 3 (NEURL3), a putative E3 ligase, in a manner that requires the involvement of innate immune sensing but is independent of the IFN action. Furthermore, we showed that NEURL3 inhibited HCV infection while it had little effect on other RNA viruses, including Zika virus (ZIKV), dengue virus (DENV), and vesicular stomatitis virus (VSV). Mechanistic studies demonstrated that NEURL3 inhibited HCV assembly by directly binding HCV envelope glycoprotein E1 to interfere with the E1/E2 heterodimerization, an important prerequisite for virion morphogenesis. Finally, we showed that knockout of NEURL3 significantly enhanced HCV infection. In summary, we identified NEURL3 as a novel inducible antiviral host factor that suppresses HCV assembly. Our results not only shed new insight into how host innate immunity acts against HCV but also revealed a new important biological function for NEURL3.IMPORTANCE The exact biological function of NEURL3, a putative E3 ligase, remains largely unknown. In this study, we found that NEURL3 could be upregulated upon HCV infection in a manner dependent on pattern recognition receptor-mediated innate immune response. NEURL3 inhibits HCV assembly by directly binding viral E1 envelope glycoprotein to disrupt its interaction with E2, an action that requires its Neuralized homology repeat (NHR) domain but not the RING domain. Furthermore, we found that NEURL3 has a pangenotypic anti-HCV activity and interacts with E1 of genotypes 2a, 1b, 3a, and 6a but does not inhibit other closely related RNA viruses, such as ZIKV, DENV, and VSV. To our knowledge, our study is the first report to demonstrate that NEURL3 functions as an antiviral host factor. Our results not only shed new insight into how host innate immunity acts against HCV, but also revealed a new important biological function for NEURL3.
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Antivirais/farmacologia , Hepatite C/prevenção & controle , Imunidade Inata/imunologia , Infecções por Vírus de RNA/virologia , Ubiquitina-Proteína Ligases/farmacologia , Proteínas do Envelope Viral/antagonistas & inibidores , Vírus da Dengue/efeitos dos fármacos , Células HEK293 , Hepacivirus/classificação , Hepacivirus/genética , Hepacivirus/imunologia , Hepatite C/imunologia , Hepatite C/virologia , Humanos , Infecções por Vírus de RNA/tratamento farmacológico , Infecções por Vírus de RNA/imunologia , Vírus de RNA/imunologia , Vírus da Estomatite Vesicular Indiana/efeitos dos fármacos , Proteínas do Envelope Viral/imunologia , Proteínas do Envelope Viral/metabolismo , Montagem de Vírus , Zika virus/efeitos dos fármacosRESUMO
Ebola virus (EBOV) causes severe hemorrhagic fever in humans and other primates with a high case fatality rate. No approved drug or vaccine of EBOV is available, which necessitates better understanding of the virus life cycle. Studies on EBOV have been hampered because experimentations involving live virus are restricted to biosafety level 4 (BSL4) laboratories. The EBOV minigenome system has provided researchers with the opportunity to study EBOV under BSL2 conditions. Here, we developed a novel EBOV minigenome replicon which, to our knowledge, is the first EBOV cell culture system that can stably replicate and transcribe the EBOV minigenome. The minigenomic RNA harboring a Gaussia luciferase and hygromycin-resistant marker can replicate for months in a helper cell stably expressing viral nucleoprotein (NP), viral protein 35 (VP35), VP30, and L proteins. Quantification of viral RNA (vRNA), cRNA, and mRNA levels of the EBOV minigenome demonstrated that the stable EBOV replicon had much-more-active minigenome replication than previously developed transient-transfection-based EBOV minigenome systems, which recapitulate viral primary transcription more than genome replication. Interestingly, minigenome replication in the stable EBOV replicon cells was insensitive to interferon treatment or RNA interference. Moreover, RNase digestion of the replicon cell lysates revealed the remarkably stable nature of the EBOV minigenomic vRNA ribonucleoprotein complex, which may help improve understanding of EBOV persistence in convalescent patients.IMPORTANCE The scope and severity of the recent Ebola outbreak in Western Africa justified a more comprehensive investigation of the causative risk group 4 agent Ebola virus (EBOV). Study of EBOV replication and antiviral development can be facilitated by developing a cell culture system that allows experimentation under biosafety level 2 conditions. Here, we developed a novel stable EBOV minigenome replicon which, to our knowledge, is the first EBOV cell culture system that can stably replicate and transcribe the EBOV minigenome. The replicon system had more-active genome replication than previously developed transient-transfection-based EBOV minigenome systems, providing a convenient surrogate system to study EBOV replication. Furthermore, self-replicating minigenomic vRNA in the replicon cells displayed strong stability in response to interferon treatment, RNA silencing, and RNase digestion, which may provide an explanation for the persistence of EBOV in survivors.
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Natural competence is the principal driver of streptococcal evolution. While acquisition of new traits could facilitate rapid fitness improvement for bacteria, entry into the competent state is a highly orchestrated event, involving an interplay between various pathways. We present a new type of competence-predation coordination mechanism in Streptococcus sanguinis. Unlike other streptococci that mediate competence through the ComABCDE regulon, several key components are missing in the S. sanguinis ComCDE circuitry. We assembled two synthetic biology devices linking competence-stimulating peptide (CSP) cleavage and export with a quantifiable readout to unravel the unique features of the S. sanguinis circuitry. Our results revealed the ComC precursor cleavage pattern and the two host ABC transporters implicated in the export of the S. sanguinis CSP. Moreover, we discovered a ComCDE-dependent bacteriocin locus. Overall, this study presents a mechanism for commensal streptococci to maximize transformation outcome in a fluid environment through extensive circuitry rewiring.
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Bacteriocinas , Streptococcus sanguis , Streptococcus sanguis/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sinais (Psicologia) , Bacteriocinas/metabolismo , PeptídeosRESUMO
Multimodal vision-language (VL) learning has noticeably pushed the tendency toward generic intelligence owing to emerging large foundation models. However, tracking, as a fundamental vision problem, surprisingly enjoys less bonus from recent flourishing VL learning. We argue that the reasons are two-fold: the lack of large-scale vision-language annotated videos and ineffective visionlanguage interaction learning of current works. These nuisances motivate us to design more effective vision-language representation for tracking, meanwhile constructing a large database with language annotation for model learning. Particularly, in this paper, we first propose a general attribute annotation strategy to decorate videos in six popular tracking benchmarks, which contributes a largescale vision-language tracking database with more than 23,000 videos. We then introduce a novel framework to improve tracking by learning a unified-adaptive VL representation, where the cores are the proposed asymmetric architecture search and modality mixer (ModaMixer). To further improve VL representation, we introduce a contrastive loss to align different modalities. To thoroughly evidence the effectiveness of our method, we integrate the proposed framework on three tracking methods with different designs, i.e., the CNNbased SiamCAR [1], the Transformer-based OSTrack [2], and the hybrid structure TransT [3]. The experiments demonstrate that our framework can significantly improve all baselines on six benchmarks. Besides empirical results, we theoretically analyze our approach to show its rationality. By revealing the potential of VL representation, we expect the community to divert more attention to VL tracking and hope to open more possibilities for future tracking with diversified multimodal messages.
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The experiments were conducted in the Tibetan plateau environment, and the sewage treatment conditions were designed with ultraviolet (UV) irradiation for 5 min, 10 min, 30 min, and 180 min. The Illumina MiSeq high-throughput sequencing technology was used to analyze the microbiological and metabolomic patterns of the plateau sewage treatment at the experimental scale, and then the response mechanisms of microbial and nitrogen metabolism in sewage treatment were explored. The abundance of metabolism at the first level and global and overview maps at the second level were higher in the plateau environment than in other regions. The KEGG pathway shows the effect of UV on nitrogen metabolism and its aptitude to improving or inhibit it. The two main nitrogen removal processes are nitrification and dissimilatory nitrate reduction. This study reveals the response of activated sludge to UV radiation in a plateau environment from microbiological and metabolomic perspectives, providing ideas and perspectives for the study of water treatment system methods, as well as laying a valuable theoretical foundation for the enhancement of plateau sewage treatment capacity.
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Esgotos , Raios Ultravioleta , Esgotos/microbiologia , Reatores Biológicos/microbiologia , Nitrificação , Nitrogênio/metabolismo , DesnitrificaçãoRESUMO
Discovering hidden pattern from imbalanced data is a critical issue in various real-world applications. Existing classification methods usually suffer from the limitation of data especially for minority classes, and result in unstable prediction and low performance. In this paper, a deep generative classifier is proposed to mitigate this issue via both model perturbation and data perturbation. Specially, the proposed generative classifier is derived from a deep latent variable model where two variables are involved. One variable is to capture the essential information of the original data, denoted as latent codes, which are represented by a probability distribution rather than a single fixed value. The learnt distribution aims to enforce the uncertainty of model and implement model perturbation, thus, lead to stable predictions. The other variable is a prior to latent codes so that the codes are restricted to lie on components in Gaussian Mixture Model. As a confounder affecting generative processes of data (feature/label), the latent variables are supposed to capture the discriminative latent distribution and implement data perturbation. Extensive experiments have been conducted on widely-used real imbalanced image datasets. Experimental results demonstrate the superiority of our proposed model by comparing with popular imbalanced classification baselines on imbalance classification task.
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Despite the excellent antiviral potency of direct-acting antivirals (DAAs) against hepatitis C virus (HCV), emergence of drug-resistant viral mutations remains a potential challenge. Sofobuvir (SOF), a nucleotide analog targeting HCV NS5B - RNA-dependent RNA polymerase (RdRp), constitutes a key component of many anti-HCV cocktail regimens and confers a high barrier for developing drug resistance. The serine to threonine mutation at the amino acid position 282 of NS5B (S282T) is the mostly documented SOF resistance-associated substitution (RAS), but severely hampers the virus fitness. In this study, we first developed new genotype 1b (GT1b) subgenomic replicon cells, denoted PR52D4 and PR52D9, directly from a GT1b clinical isolate. Next, we obtained SOF-resistant and replication-competent PR52D4 replicon by culturing the replicon cells in the presence of SOF. Sequencing analysis showed that the selected replicon harbored two mutations K74R and S282T in NS5B. Reverse genetics analysis showed that while PR52D4 consisting of either single mutation K74R or S282T could not replicate efficiently, the engineering of the both mutations led to a replication-competent and SOF-resistant PR52D4 replicon. Furthermore, we showed that the K74R mutation could also rescue the replication deficiency of the S282T mutation in Con1, another GT1b replicon as well as in JFH1, a GT2a replicon. Structural modeling analysis suggested that K74R might help maintain an active catalytic conformation of S282T by engaging with Y296. In conclusion, we identified the combination of two NS5B mutations S282T and K74R as a novel RAS that confers a substantial resistance to SOF while retains the HCV replication capacity.
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Antivirais/farmacologia , Farmacorresistência Viral/genética , Variação Genética , Hepacivirus/efeitos dos fármacos , Hepacivirus/genética , Replicon/genética , Sofosbuvir/farmacologia , Genótipo , Hepacivirus/isolamento & purificação , Hepatite C/virologia , Humanos , Replicon/efeitos dos fármacosRESUMO
Hepatitis C virus (HCV) remains a major human pathogen that requires better understanding of virus-host interactions. In this study, we performed a genome-wide CRISPR-Cas9 screening and identified TRIM26, an E3 ligase, as a critical HCV host factor. Deficiency of TRIM26 specifically impairs HCV genome replication. Mechanistic studies showed that TRIM26 interacts with HCV-encoded NS5B protein and mediates its K27-linked ubiquitination at residue K51, and thus promotes the NS5B-NS5A interaction. Moreover, mouse TRIM26 does not support HCV replication because of its unique six-amino acid insert that prevents its interaction with NS5B. Ectopic expression of human TRIM26 in a mouse hepatoma cell line that has been reconstituted with other essential HCV host factors promotes HCV infection. In conclusion, we identified TRIM26 as a host factor for HCV replication and a new determinant of host tropism. These results shed light on HCV-host interactions and may facilitate the development of an HCV animal model.
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Chronic hepatitis C infection is a leading cause of liver cirrhosis, which is linked to chronic hepatic inflammation. While there are multiple studies detailing the proinflammatory role of interleukin-1ß (IL-1ß) in HCV-induced inflammasome signaling, the antiviral capacity of this cytokine has not been adequately investigated in the context of HCV infection or other members of Flaviridae. Our data indicated that IL-1ß alone does not inhibit HCV replication, yet when in combination with IFN-α, it can boost the anti-HCV activity of IFN-α, which is mediated by augmented STAT1 tyrosine 701 phosphorylation. Through signaling inhibitor screening, we found that ERK2 kinase is directly linked to the enhanced activation of the STAT1 complex. Our study found that IL-1ß negatively affects ERK2 phosphorylation, which suggests that IL-1ß-mediated STAT1 tyrosine 701 phosphorylation employed kinase machinery of ERK2 other than JNK or P38 kinase. Our results identify IL-1ß as a proinflammatory cytokine possessing wide spectrum synergistic antiviral capability via enhancing IFN-α-induced interferon-stimulated genes (ISGs) expression. A more nuanced understanding of the antiviral mechanisms of this important cytokine could facilitate the development of new therapeutic options.
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Antivirais , Hepatite C , Antivirais/farmacologia , Antivirais/uso terapêutico , Hepacivirus , Humanos , Interferon-alfa/farmacologia , Interferon-alfa/uso terapêutico , Interleucina-1beta/uso terapêuticoRESUMO
Hepatitis C virus (HCV), a major causative agent of chronic hepatitis, is a positive-stranded RNA virus and has a high degree of genetic diversity due to its error-prone RNA-dependent RNA polymerase. Development of direct-acting antiviral agents (DAAs) has greatly improved the therapeutic outcome of chronic hepatitis C patients. However, naturally existing resistance-associated variants (RAVs) or occurrence of resistance-associated substitutions (RASs) in the HCV genome may impose a challenge to the long-term success of the DAA-based therapies. Genotype-3 HCV is the most difficult genotype to treat by DAAs, but the underlying molecular mechanisms remain to be explored. Here we developed a novel genotype-3a subgenomic replicon PR87A7 by screening a HCV cDNA pool amplified from a patient serum RNA. PR87A7 replicon displayed strong resistance to anti-NS3 DAAs, mainly owing to a genotype-3-specific polymorphism 168Q in NS3. Introduction of NS3 168Q into a genotype-2a JFH1 strain rendered resistance to anti-NS3 DAAs while greatly diminished the viral replication, and yet this fitness defect can be rescued by additional genotype-3-specific polymorphism. In conclusion, we developed a novel genotype-3a subgenomic replicon by a functional screening approach, and revealed genotype-3-specfic amino acid residues that confer resistance to anti-NS3 DAAs while retaining viral fitness.
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Farmacorresistência Viral , Genótipo , Hepacivirus/efeitos dos fármacos , Hepacivirus/genética , Hepatite C/virologia , Polimorfismo Genético , Replicação Viral/efeitos dos fármacos , Substituição de Aminoácidos , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Aptidão Genética , Genoma Viral , Hepacivirus/classificação , Hepatite C/tratamento farmacológico , Humanos , Filogenia , Proteínas não Estruturais Virais/genéticaRESUMO
Genotype 1b strain Con1 represents an important reference in the study of hepatitis C virus (HCV). Here, we aimed to develop an advanced infectious Con1 recombinant. We found that previously identified mutations A1226G/F1464L/A1672S/Q1773H permitted culture adaption of Con1 Core-NS5A (C-5A) recombinant containing 5'UTR and NS5B-3'UTR from JFH1 (genotype 2a), thus acquired additional mutations L725H/F886L/D2415G. C-5A containing all seven mutations (C-5A_7m) replicated efficiently in Huh7.5 and Huh7.5.1 cells and had an increased infectivity in SEC14L2-expressing Huh7.5.1 cells. Incorporation of Con1 NS5B was deleterious to C-5A_7m, however Con1 5'UTR was permissive but attenuated the virus. Nucleotides G1, A4, and G35 primarily accounted for the viral attenuation without affecting RNA translation. C-5A_7m was inhibited dose-dependently by simeprevir and daclatasvir, and substitutions at A4, A29, A34, and G35 conferred resistance to miR-122 antagonism. The novel Con1 5'UTR-NS5A recombinant, adaptive mutations, and critical nucleotides described here will facilitate future studies of HCV culture systems and virus-host interaction.