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1.
J Sci Food Agric ; 104(2): 1092-1106, 2024 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-37782112

RESUMO

BACKGROUND: Flavonoids are vital for the development of high-quality grapes and wine, and manganese deficiency decreases grape berry coloration. However, the effects and underlying mechanisms of action of manganese sulfate on grape metabolic profiles have not been adequately researched. In this study, three concentrations of manganese sulfate solutions, 0.5 µmol·L-1 (low, L), 5 µmol·L-1 (middle, M - the standard manganese concentration of Hoagland nutrient solution, control), and 1000 µmol·L-1 (high, H), were applied to the 'Cabernet Sauvignon' grapevine (Vitis vinifera L.) to explore the effect on berry composition. RESULTS: Manganese application improved manganese concentration effectively in grape organs. Furthermore, the concentrations of malvidin 3-O-(6-O-acetyl)-glucoside, malvidin 3-O-glucoside, malvidin-trans-3-O-(6-O-p-coumaryl)-glucoside, and peonidin 3-O-(6-O-acetyl)-glucoside increased significantly under H treatment. Weighted gene co-expression network analysis (WGCNA) revealed that the structural genes (VvDFR, VvUFGT, and VvOMT) of flavonoid biosynthesis were upregulated under H treatment, and their transcription levels correlated positively with malvidin- and peonidin-derived anthocyanin concentrations. CONCLUSIONS: This study suggested that manganese application regulates berry transcriptional and flavonoid metabolic profiles, providing a theoretical basis for improving the color of red grapes and wines. © 2023 Society of Chemical Industry.


Assuntos
Vitis , Vinho , Vitis/química , Flavonoides/análise , Transcriptoma , Manganês/análise , Antocianinas/análise , Vinho/análise , Metaboloma , Glucosídeos/análise , Frutas/química
2.
Environ Geochem Health ; 46(3): 105, 2024 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-38441743

RESUMO

The extensive use of organic amine pesticides (OAPs) in agricultural practices has resulted in the contamination of water environments, posing threats to ecosystems and human health. This study focused on the Xiang River (XR), a representative drinking water source, as the research area to investigate the occurrence characteristics of 34 OAPs. Diphenylamine emerged as the most prevalent OAP in surface water due to industrial and agricultural activities, while cycloate dominated in sediments due to cumulative effects. Generally, the concentration of OAPs in a mixed tap water sample was lower than those in surface water samples, indicating OAPs can be removed by water plants to a certain extent. The water-sediment distribution coefficients (kd) of ΣOAPs were much less than 1 L/g, the majority of OAPs maintained relatively high concentrations in water samples instead of accumulating in sediments. Furthermore, risk assessment revealed that carbofuran showed a moderate risk to the aquatic environment, with a risk quotient of 0.23, while other OAPs presented minor risks. This study provided crucial insights for regional pesticide management and control in the XR basin, emphasizing the importance of implementing strategies to minimize the release of OAPs into the environment and protect human health.


Assuntos
Água Potável , Praguicidas , Humanos , Aminas , Ecossistema , Rios , China , Medição de Risco
3.
New Phytol ; 240(4): 1433-1448, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37668229

RESUMO

The transcription of photosynthesis genes in chloroplasts is largely mediated by the plastid-encoded RNA polymerase (PEP), which resembles prokaryotic-type RNA polymerases, but with plant-specific accessory subunits known as plastid transcriptionally active chromosome proteins (pTACs) or PEP-associated proteins (PAPs). However, whether additional factors are involved in the biogenesis of PEP complexes remains unknown. Here, we investigated the function of an essential gene, PALE CRESS (PAC), in the accumulation of PEP complexes in chloroplasts. We established that an Arabidopsis leaf variegation mutant, variegated 6-1 (var6-1), is a hypomorphic allele of PAC. Unexpectedly, we revealed that a fraction of VAR6/PAC is associated with thylakoid membranes, where it interacts with PEP complexes. The accumulation of PEP complexes is defective in both var6-1 and the null allele var6-2. Further protein interaction assays confirmed that VAR6/PAC interacts directly with the PAP2/pTAC2 and PAP3/pTAC10 subunits of PEP complexes. Moreover, we generated viable hypomorphic alleles of the essential gene PAP2/pTAC2, and revealed a genetic interaction between PAC and PAP2/pTAC2 in photosynthesis gene expression and PEP complex accumulation. Our findings establish that VAR6/PAC affects PEP complex accumulation through interactions with PAP2/pTAC2 and PAP3/pTAC10, and provide new insights into the accumulation of PEP and chloroplast development.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Brassicaceae , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Brassicaceae/metabolismo , Proteínas de Cloroplastos/metabolismo , Cloroplastos/metabolismo , RNA Polimerases Dirigidas por DNA/metabolismo , Regulação da Expressão Gênica de Plantas , Mutação/genética , Plastídeos/genética , Fatores de Transcrição/metabolismo
4.
J Sci Food Agric ; 103(10): 4838-4849, 2023 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-36916448

RESUMO

BACKGROUND: In current vineyards, potassium dihydrogen phosphate (KH2 PO4 ) is a common foliar fertilizer with the lowest salt index. It is employed to improve the transportation and distribution of grape photosynthetic products, but the mechanism of its effect on fruit flavonoid synthesis is unclear. RESULTS: This study investigated the effects of foliar spraying of KH2 PO4 at different developmental stages (1 week before veraison; the end of veraison (EV)) on flavonoid metabolites and related gene expression of 'Cabernet Sauvignon' grape for two consecutive vintages. High-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry technology was used to identify 6 flavan-3-ols, 11 flavonols, and 16 anthocyanins. KH2 PO4 influenced anthocyanins content, especially when applied at the EV stage, the content of anthocyanins was significantly higher than that of the control. Further, quantitative polymerase chain reaction analysis showed that KH2 PO4 treatment applied at the EV stage can increase the expression of anthocyanin synthesis genes and accelerate anthocyanin synthesis. In particular, the expression of VviGST in EV treatment was significantly higher than that of the control during the development process. CONCLUSION: These findings have enhanced our understanding of the effect of KH2 PO4 treatment on grape flavonoids. Among them, EV treatment can significantly increase anthocyanins content. © 2023 Society of Chemical Industry.


Assuntos
Flavonoides , Vitis , Flavonoides/análise , Vitis/química , Antocianinas/análise , Fosfatos/análise , Frutas/química
5.
J Immunol ; 204(2): 394-407, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31826939

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) has caused tremendous economic losses in the swine industry since its emergence in the late 1980s. PRRSV exploits various strategies to evade immune responses and establish chronic persistent infections. Suppressor of cytokine signaling (SOCS) 1, a member of the SOCS family, is a crucial intracellular negative regulator of innate immunity. In this study, it was shown that SOCS1 can be co-opted by PRRSV to evade host immune responses, facilitating viral replication. It was observed that PRRSV induced SOCS1 production in porcine alveolar macrophages, monkey-derived Marc-145 cells, and porcine-derived CRL2843-CD163 cells. SOCS1 inhibited the expression of IFN-ß and IFN-stimulated genes, thereby markedly enhancing PRRSV replication. It was observed that the PRRSV N protein has the ability to upregulate SOCS1 production and that nuclear localization signal-2 (NLS-2) is essential for SOCS1 induction. Moreover, SOCS1 upregulation was dependent on p38/AP-1 and JNK/AP-1 signaling pathways rather than classical type I IFN signaling pathways. In summary, to our knowledge, the findings of this study uncovered the molecular mechanism that underlay SOCS1 induction during PRRSV infection, providing new insights into viral immune evasion and persistent infection.


Assuntos
Macrófagos Alveolares/imunologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Proteína 1 Supressora da Sinalização de Citocina/metabolismo , Fator de Transcrição AP-1/metabolismo , Animais , Linhagem Celular , Haplorrinos , Evasão da Resposta Imune , Interferons/genética , MAP Quinase Quinase 4/metabolismo , Macrófagos Alveolares/virologia , Sinais de Localização Nuclear/genética , Proteínas do Nucleocapsídeo/genética , Proteínas do Nucleocapsídeo/metabolismo , Transdução de Sinais , Proteína 1 Supressora da Sinalização de Citocina/genética , Suínos , Fator de Transcrição AP-1/genética , Replicação Viral , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
6.
J Sci Food Agric ; 102(13): 6100-6111, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35474458

RESUMO

BACKGROUND: Vitis vinifera L. cv. Syrah grapevines in most Chinese viticulture regions generally have compact clusters that increase the susceptibility to diseases and inhibit coloration of the inner berries. Gibberellic acid (GA3 ) is a plant growth regulator that is widely used during grape cultivation to elongate the rachis, control fruit set, and decrease cluster compactness. In this study, Syrah grapevines were treated with GA3 before flowering in 2019 and 2020 to determine the optimal GA3 treatment concentrations and times for decreasing bunch compactness, while minimizing the negative effects on the wine grape cluster weight. RESULTS: Pre-flowering GA3 applications at 3, 5, and 7 mg L-1 , especially treatment at 20 days before flowering, decreased Syrah grape bunch compactness by decreasing the fruit set rate and promoting bunch elongation, with minimal adverse effects on the healthy grape cluster weight in both years. The 7 mg L-1 GA3 treatment at 20 days before flowering significantly increased reducing sugar, total phenolic, tannin, and total anthocyanin contents of Syrah grape berries in 2019 and 2020. Moreover, high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry, hierarchical cluster, and principal component analysis results indicated GA3 applications before flowering (3, 5, and 7 mg L-1 ) significantly affected the accumulation of different anthocyanins in Syrah grape berries. Notably, the application of 7 mg L-1 GA3 at 20 days before flowering resulted in the highest anthocyanin content. CONCLUSION: Pre-flowering gibberellin application can decrease bunch compactness and improve the quality of Syrah grape berries. These findings reflect the potential utility of gibberellin treatments for decreasing cluster compactness and increasing the quality of wine grapes. © 2022 Society of Chemical Industry.


Assuntos
Vitis , Vinho , Antocianinas/análise , Flavonoides/análise , Frutas/química , Giberelinas/farmacologia , Fenótipo , Vitis/química , Vinho/análise
7.
J Digit Imaging ; 34(3): 691-704, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-34080105

RESUMO

Age-related macular degeneration (AMD) is one of the leading causes of irreversible blindness and is characterized by fluid-related accumulations such as intra-retinal fluid (IRF), subretinal fluid (SRF), and pigment epithelial detachment (PED). Spectral-domain optical coherence tomography (SD-OCT) is the primary modality used to diagnose AMD, yet it does not have algorithms that directly detect and quantify the fluid. This work presents an improved convolutional neural network (CNN)-based architecture called RetFluidNet to segment three types of fluid abnormalities from SD-OCT images. The model assimilates different skip-connect operations and atrous spatial pyramid pooling (ASPP) to integrate multi-scale contextual information; thus, achieving the best performance. This work also investigates between consequential and comparatively inconsequential hyperparameters and skip-connect techniques for fluid segmentation from the SD-OCT image to indicate the starting choice for future related researches. RetFluidNet was trained and tested on SD-OCT images from 124 patients and achieved an accuracy of 80.05%, 92.74%, and 95.53% for IRF, PED, and SRF, respectively. RetFluidNet showed significant improvement over competitive works to be clinically applicable in reasonable accuracy and time efficiency. RetFluidNet is a fully automated method that can support early detection and follow-up of AMD.


Assuntos
Degeneração Macular , Tomografia de Coerência Óptica , Humanos , Redes Neurais de Computação , Retina/diagnóstico por imagem , Líquido Sub-Retiniano/diagnóstico por imagem
8.
Child Youth Serv Rev ; 118: 105359, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32836604

RESUMO

This study examined the accessibility, affordability, accountability, sustainability, and social justice of early childhood education (ECE) services in Shenzhen, China, using Li et al.'s (2017) '3A2S' framework. Government documents and secondary data during the past decade were collected and evaluated. The results indicated that: (1) the ECE services have improved in the dimensions of accessibility, affordability, accountability, sustainability, and social justice; (2) more efforts should be made in increasing fiscal budget into ECE services and ensuring the quality of the ECE services; and (3) the government needs to take up more responsibilities to strike a balance between market force and governmental regulation. Implications and suggestions are also included.

9.
J Virol ; 92(12)2018 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-29618647

RESUMO

Once infected by viruses, cells can detect pathogen-associated molecular patterns (PAMPs) on viral nucleic acid by host pattern recognition receptors (PRRs) to initiate the antiviral response. Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of porcine reproductive and respiratory syndrome (PRRS), characterized by reproductive failure in sows and respiratory diseases in pigs of different ages. To date, the sensing mechanism of PRRSV has not been elucidated. Here, we reported that the pseudoknot region residing in the 3' untranslated regions (UTR) of the PRRSV genome, which has been proposed to regulate RNA synthesis and virus replication, was sensed as nonself by retinoic acid-inducible gene I (RIG-I) and Toll-like receptor 3 (TLR3) and strongly induced type I interferons (IFNs) and interferon-stimulated genes (ISGs) in porcine alveolar macrophages (PAMs). The interaction between the two stem-loops inside the pseudoknot structure was sufficient for IFN induction, since disruption of the pseudoknot interaction powerfully dampened the IFN induction. Furthermore, transfection of the 3' UTR pseudoknot transcripts in PAMs inhibited PRRSV replication in vitro Importantly, the predicted similar structures of other arterivirus members, including equine arteritis virus (EAV), lactate dehydrogenase-elevating virus (LDV), and simian hemorrhagic fever virus (SHFV), also displayed strong IFN induction activities. Together, in this work we identified an innate recognition mechanism by which the PRRSV 3' UTR pseudoknot region served as PAMPs of arteriviruses and activated innate immune signaling to produce IFNs that inhibit virus replication. All of these results provide novel insights into innate immune recognition during virus infection.IMPORTANCE PRRS is the most common viral disease in the pork industry. It is caused by PRRSV, a positive single-stranded RNA virus, whose infection often leads to persistent infection. To date, it is not yet clear how PRRSV is recognized by the host and what is the exact mechanism of IFN induction. Here, we investigated the nature of PAMPs on PRRSV and the associated PRRs. We found that the 3' UTR pseudoknot region of PRRSV, which has been proposed to regulate viral RNA synthesis, could act as PAMPs recognized by RIG-I and TLR3 to induce type I IFN production to suppress PRRSV infection. This report is the first detailed description of pattern recognition for PRRSV, which is important in understanding the antiviral response of arteriviruses, especially PRRSV, and extends our knowledge on virus recognition.


Assuntos
Proteína DEAD-box 58/genética , Moléculas com Motivos Associados a Patógenos/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Dobramento de RNA/genética , Receptor 3 Toll-Like/genética , Regiões 3' não Traduzidas/genética , Animais , Linhagem Celular , Chlorocebus aethiops , Genoma Viral/genética , Helicase IFIH1 Induzida por Interferon/genética , Interferon-alfa/imunologia , Interferon beta/imunologia , Sequências Repetidas Invertidas/genética , Moléculas com Motivos Associados a Patógenos/metabolismo , Síndrome Respiratória e Reprodutiva Suína/imunologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Interferência de RNA , RNA Interferente Pequeno/genética , Sus scrofa , Suínos , Doenças dos Suínos/virologia , Receptor 7 Toll-Like/genética
10.
Breast J ; 25(4): 712-715, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31079421

RESUMO

The authors report the case of a 56-year-old woman with mammary sparganosis due to infection with a plerocercoid tapeworm larva of Spirometra mansoni. Magnetic resonance imaging revealed an area of heterogeneous density in outer upper quadrant of the right breast, with a high likelihood of malignancy. During surgery for the removal of a granuloma, the parasite was discovered and excised. The authors review the pathological and imaging features of mammary sparganosis.


Assuntos
Doenças Mamárias/parasitologia , Doenças Mamárias/cirurgia , Esparganose/parasitologia , Esparganose/cirurgia , Spirometra/patogenicidade , Animais , Doenças Mamárias/diagnóstico por imagem , Feminino , Humanos , Imageamento por Ressonância Magnética , Pessoa de Meia-Idade , Esparganose/diagnóstico por imagem , Ultrassonografia Mamária
11.
Am J Hum Genet ; 96(4): 597-611, 2015 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-25839328

RESUMO

Esophageal squamous cell carcinoma (ESCC) is one of the most common cancers worldwide and the fourth most lethal cancer in China. However, although genomic studies have identified some mutations associated with ESCC, we know little of the mutational processes responsible. To identify genome-wide mutational signatures, we performed either whole-genome sequencing (WGS) or whole-exome sequencing (WES) on 104 ESCC individuals and combined our data with those of 88 previously reported samples. An APOBEC-mediated mutational signature in 47% of 192 tumors suggests that APOBEC-catalyzed deamination provides a source of DNA damage in ESCC. Moreover, PIK3CA hotspot mutations (c.1624G>A [p.Glu542Lys] and c.1633G>A [p.Glu545Lys]) were enriched in APOBEC-signature tumors, and no smoking-associated signature was observed in ESCC. In the samples analyzed by WGS, we identified focal (<100 kb) amplifications of CBX4 and CBX8. In our combined cohort, we identified frequent inactivating mutations in AJUBA, ZNF750, and PTCH1 and the chromatin-remodeling genes CREBBP and BAP1, in addition to known mutations. Functional analyses suggest roles for several genes (CBX4, CBX8, AJUBA, and ZNF750) in ESCC. Notably, high activity of hedgehog signaling and the PI3K pathway in approximately 60% of 104 ESCC tumors indicates that therapies targeting these pathways might be particularly promising strategies for ESCC. Collectively, our data provide comprehensive insights into the mutational signatures of ESCC and identify markers for early diagnosis and potential therapeutic targets.


Assuntos
Carcinoma de Células Escamosas/genética , Citidina Desaminase/genética , Neoplasias Esofágicas/genética , Predisposição Genética para Doença/genética , Genoma Humano/genética , Mutação/genética , Fosfatidilinositol 3-Quinases/genética , Transdução de Sinais/genética , Desaminase APOBEC-1 , Análise de Variância , Sequência de Bases , Proteína de Ligação a CREB/genética , Linhagem Celular Tumoral , China , Classe I de Fosfatidilinositol 3-Quinases , Variações do Número de Cópias de DNA/genética , Carcinoma de Células Escamosas do Esôfago , Técnicas de Silenciamento de Genes , Humanos , Immunoblotting , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Proteínas com Domínio LIM/genética , Ligases , Dados de Sequência Molecular , Receptores Patched , Receptor Patched-1 , Complexo Repressor Polycomb 1/genética , Proteínas do Grupo Polycomb/genética , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Superfície Celular/genética , Análise de Sequência de DNA , Sais de Tetrazólio , Tiazóis , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor/genética , Ubiquitina Tiolesterase/genética , Ubiquitina-Proteína Ligases/genética
14.
Virus Genes ; 54(1): 86-97, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29090410

RESUMO

Recently, NADC30-like porcine reproductive and respiratory syndrome viruses (PRRSVs), which are genetically similar to the NADC30 strain isolated in the United States of America in 2008, have become prevalent in China. Here, a novel variant PRRSV strain named HNhx was successfully isolated on porcine alveolar macrophages from Henan province and the full-length genome sequence was determined. Phylogenetic analysis indicated that HNhx strain was classified into the NADC30-like PRRSV subgroup, in which all the strains had the unique discontinuous 131-amino acid deletion relative to that of the nonstructural protein 2 (Nsp2) of the VR2332 strain. Genetically, HNhx shared 92.9% nucleotide similarity to NADC30. Furthermore, HNhx strain contained extensive amino acid mutations in GP5. In particular, the S32H, N33D, D34N, and S36G variations resulted in that HNhx lost all the putative N-linked glycosylation sites at amino acid positions 30, 32, 33, 34, and 35. Recombination analysis revealed that HNhx was the result of recombination between the NADC30 strain and the highly pathogenic PRRSV vaccine strain circulating in China in Nsp4 (nt 5261) to Nsp9 (nt 7911). The novel genome data of HNhx will be helpful for understanding the evolution and epidemiology of PRRSV in China.


Assuntos
Variação Genética , Genótipo , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/isolamento & purificação , Animais , Células Cultivadas , China , Genoma Viral , Macrófagos Alveolares/virologia , Filogenia , Síndrome Respiratória e Reprodutiva Suína/virologia , Recombinação Genética , Análise de Sequência de DNA , Deleção de Sequência , Homologia de Sequência do Ácido Nucleico , Suínos , Sequenciamento Completo do Genoma
15.
BMC Biotechnol ; 17(1): 77, 2017 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-29121904

RESUMO

BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) exhibits a highly restricted tropism for cells of the monocyte-macrophage lineage, utilizing porcine CD163 (pCD163) as an indispensable cellular receptor for infection. Transfection the gene of pCD163 into several non-permissive cell lines followed by protein expression confers susceptibility to PRRSV. A lack of specialized porcine antibody tools for use with existing porcine-derived primary cells and cell lines has hampered studies of both PRRSV pathogenesis and virus triggering of immune response cascades. Therefore, we constructed PRRSV-susceptible murine alveolar macrophage-derived MH-S and peritoneal macrophage-like RAW264.7 cell lines by achieving pCD163 cell surface expression in these cells. We then evaluated PRRSV susceptibility and cytokine expression patterns induced upon PRRSV infection of these pCD163-expressing cell lines. RESULTS: Growth of MH-SCD163 and RAW264.7CD163 cells was indistinguishable from growth of un-transfected parental cell lines. Meanwhile, various stages of the PRRSV replication cycle, including viral particle attachment, internalization, disassembly and infection were confirmed in both pCD163-transfected cell lines. Analysis of PRRSV replication using immunofluorescence staining of virus and viral titration of cell lysates demonstrated that both MH-SCD163 and RAW264.7CD163 cells supported replication of various genotype 2 PRRSV isolates. Moreover, PRRSV replication in MH-SCD163 cells was similar to that observed in porcine alveolar macrophages (PAMs) and was more efficient than in RAW264.7CD163 cells. However, peak virus titers in MH-SCD163 cells were attained at 60 h post-infection (pi) versus 48 hpi in PAMs. Analysis of cytokine expression showed that post-PRRSV infection, mRNA expression patterns of anti-inflammatory cytokines (IL-4 and IL-10) and pro-inflammatory cytokines (TNF-α and IFN-γ) in MH-SCD163 cells were more similar to those observed in PAMs versus levels in RAW264.7CD163 cells. CONCLUSIONS: MH-S and RAW264.7 cells were not susceptible to PRRSV infection until transfection and subsequent expression of pCD163 were achieved in these cell lines. The PRRSV-susceptible MH-SCD163 cell line efficiently supported viral replication of various genotype 2 PRRSV isolates and exhibited similar cytokine expression patterns as observed in PAMs. In conclusion, this work describes the development of new tools to further understand PRRSV pathogenesis and immune response mechanisms to PRRSV infection.


Assuntos
Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Macrófagos/citologia , Macrófagos/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/crescimento & desenvolvimento , Receptores de Superfície Celular/metabolismo , Cultura de Vírus/métodos , Animais , Linhagem Celular , Proliferação de Células , Macrófagos/metabolismo , Camundongos , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Vírus da Síndrome Respiratória e Reprodutiva Suína/metabolismo , Células RAW 264.7 , Suínos
16.
Microb Pathog ; 111: 14-21, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28804020

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) is acknowledged a fulminating infectious pathogen affecting the pig farming industry, and current vaccines and drugs could hardly inhibit this virus. The 2', 5'-oligoadenylate synthetase (OASs) have antiviral activities, but the role(s) played by porcine OAS2 in protection against PRRSV infection are unknown. Here we found that endogenous expression of the porcine OAS2 gene could be promoted by interferon (IFN)-beta or PRRSV infection in porcine alveolar macrophages. Knockdown of porcine OAS2 led to increases in PRRSV replication, and OAS2 expression suppressed replication of PRRSV in a retinoic acid inducible gene I (RIG-I)-dependent manner, anti-PRRSV activity of porcine OAS2 would be lost if RNase L and OAS2 were both silenced. This discovery illustrates a pathway that porcine OAS2 responses to host anti-PRRSV function.


Assuntos
2',5'-Oligoadenilato Sintetase/antagonistas & inibidores , Antivirais/farmacologia , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Vírus da Síndrome Respiratória e Reprodutiva Suína/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , 2',5'-Oligoadenilato Sintetase/genética , 2',5'-Oligoadenilato Sintetase/metabolismo , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Endorribonucleases/genética , Expressão Gênica , Perfilação da Expressão Gênica , Técnicas de Silenciamento de Genes , Interações Hospedeiro-Patógeno , Interferon beta , Pulmão , Macrófagos Alveolares/metabolismo , Macrófagos Alveolares/virologia , RNA Interferente Pequeno/metabolismo , Suínos
17.
Virus Genes ; 52(1): 91-8, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26743534

RESUMO

Porcine epidemic diarrhea virus (PEDV) has caused devastating impact on pig-rearing industry in China and current vaccine is not effective against the circulating PEDV variants. In the present study, the full-length genome sequence from a PEDV isolate (CH/HNQX-3/14) was determined. The complete genome sequence analysis showed that the CH/HNQX-3/14 possessed unique deletion regions in the S and ORF3 genes. It was identified as a recombinant strain using phylogenetic analysis and recombination detection program. Further analyses of the full-length sequence suggest that CH/HNQX-3/14 is a natural recombinant between the attenuated vaccine strains (CV777 and DR13) and circulating wild-type strain (CH/ZMDZY/11). The recombination occurred not only in structural protein-coding region (S1 and N genes) but also in non-structural protein-coding region (replicases 1a and ORF3 genes). These results provided new evidence that PEDV strains circulating in China underwent recombination between vaccine and field strains, suggesting that recombination contributes to the genetic diversity of PEDV. Our findings provide valuable information on PEDV evolution and underscore the need for ongoing surveillance of this economically important swine disease.


Assuntos
Genoma Viral , Vírus da Diarreia Epidêmica Suína/genética , Vírus Reordenados/genética , Animais , Sequência de Bases , China , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Variação Genética , Filogenia , Vírus da Diarreia Epidêmica Suína/classificação , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , RNA Viral , Recombinação Genética , Análise de Sequência de RNA , Suínos , Vacinas Virais/genética
18.
BMC Vet Res ; 11: 195, 2015 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-26260476

RESUMO

BACKGROUND: Avian hepatitis E virus (HEV) is the main causative agent of big liver and spleen disease, as well as hepatitis-splenomegaly syndrome in chickens. To date, conventional reverse transcriptase polymerase chain reaction (RT-PCR) and nested RT-PCR methods have been used for the diagnosis of avian HEV infection in chickens. However, these assays are time consuming, inconvenient, and cannot detect the virus quantitatively. In this study, a rapid and sensitive SYBR Green real-time RT-PCR assay was developed to detect avian HEV RNA quantitatively in serum, liver, spleen, and fecal samples from chickens. RESULTS: Based on the sequence of the most conserved HEV gene, ORF3, the primers for the assay were designed, and the standard plasmid was constructed. The detection limit of the assay was shown to be 10 copies/µl of standard plasmid/reaction, with a corresponding cycle-threshold value of 29.3. The standard curve exhibited a dynamic linear range across at least 7 log units of DNA copy number. The specificity and reproducibility of this assay was high, showing that the assay detected avian HEV RNA specifically and with little variability. Compared to conventional RT-PCR, the current assay is more sensitive for detecting avian HEV in serum, liver, spleen, and fecal samples from chickens. CONCLUSIONS: A rapid, specific, and reproducible SYBR Green real-time RT-PCR assay was developed for the diagnosis of avian HEV infection in chickens. This assay can accurately detect avian HEV RNA in serum, liver, spleen, and fecal samples with more sensitivity than conventional RT-PCR.


Assuntos
Hepatite Viral Animal/virologia , Hepevirus/isolamento & purificação , Doenças das Aves Domésticas/virologia , Infecções por Vírus de RNA/veterinária , Animais , Benzotiazóis , Galinhas , Diaminas , Regulação Viral da Expressão Gênica/fisiologia , Hepatite Viral Animal/diagnóstico , Compostos Orgânicos , Doenças das Aves Domésticas/diagnóstico , Quinolinas , Infecções por Vírus de RNA/diagnóstico , Infecções por Vírus de RNA/virologia , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sensibilidade e Especificidade , Coloração e Rotulagem/métodos , Proteínas Virais/genética , Proteínas Virais/metabolismo
19.
Molecules ; 20(12): 22767-80, 2015 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-26703539

RESUMO

Yan73, a teinturier (dyer) grape variety in China, is one of the few Vitis vinifera cultivars with red-coloured berry flesh. To examine the tissue-specific expression of genes associated with berry colour in Yan73, we analysed the differential accumulation of anthocyanins in the skin and flesh tissues of two red-skinned grape varieties with either red (Yan73) or white flesh (Muscat Hamburg) based on HPLC-MS analysis, as well as the differential expression of 18 anthocyanin biosynthesis genes in both varieties by quantitative RT-PCR. The results revealed that the transcripts of GST, OMT, AM3, CHS3, UFGT, MYBA1, F3'5'H, F3H1 and LDOX were barely detectable in the white flesh of Muscat Hamburg. In particular, GST, OMT, AM3, CHS3 and F3H1 showed approximately 50-fold downregulation in the white flesh of Muscat Hamburg compared to the red flesh of Yan73. A correlation analysis between the accumulation of different types of anthocyanins and gene expression indicated that the cumulative expression of GST, F3'5'H, LDOX and MYBA1 was more closely associated with the acylated anthocyanins and the 3'5'-OH anthocyanins, while OMT and AM3 were more closely associated with the total anthocyanins and methoxylated anthocyanins. Therefore, the transcripts of OMT, AM3, GST, F3'5'H, LDOX and MYBA1 explained most of the variation in the amount and composition of anthocyanins in skin and flesh of Yan73. The data suggest that the specific localization of anthocyanins in the flesh tissue of Yan73 is most likely due to the tissue-specific expression of OMT, AM3, GST, F3'5'H, LDOX and MYBA1 in the flesh.


Assuntos
Antocianinas/genética , Regulação da Expressão Gênica de Plantas/genética , Vitis/genética , China , Frutas/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética
20.
Foods ; 13(8)2024 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-38672880

RESUMO

Green leaf volatiles (GLVs) are important in giving grape a fresh and green aroma. But the changes in GLVs during the phenological development of grapevines are not well known. This study analyzed the GLVs and transcription levels of associated biosynthetic genes in six grape species from the Loess Plateau region at five stages of maturation. Thirteen GLVs were detected, showing unique patterns for each grape type at various growth phases. The primary components in six grapes were (E)-2-hexenal, (E)-2-hexen-1-ol, and hexanal. With the exception of Cabernet Franc in 2019, the overall GLV contents of the six types generally increased during growth and development, peaking or stabilizing at harvest. And Sauvignon Blanc, Cabernet Gernischt, and Cabernet Sauvignon exhibited higher total contents among the varieties. PLS-DA analysis revealed 3-hexenal's high VIP scores across two years, underscoring its critical role in grape variety classification. Correlation analysis revealed a strong positive correlation between the levels of hexanal, 1-hexanol, (E)-2-hexen-1-ol, (Z)-3-hexenyl acetate, nonanal, and (E, E)-2,6-nonadienal and the expression of VvHPL and VvAAT genes in the LOX-HPL pathway. Specifically, VvHPL emerges as a potential candidate gene responsible for species-specific differences in GLV compounds. Comprehending the changing patterns in the biosynthesis and accumulation of GLVs offers viticulturists and enologists the opportunity to devise targeted strategies for improving the aromatic profile of grapes and wines.

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