Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 68
Filtrar
1.
Molecules ; 26(2)2021 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-33466694

RESUMO

This study was focused on extraction, radical scavenging activities, and chemical composition identification of total flavonoids in sunflower (Helianthus annuus L.) receptacles (TFSR). We investigated the optimal extract parameters of TFSR using response surface methodology. The highest yield of TFSR was 1.04% with the ethanol concentration 58%, the material-to-liquid ratio 1:20 (v/w), the extraction time 2.6 h, and the extraction temperature 67 °C. The results of radical scavenging activities showed that ethyl acetate fraction (EAF) was the strongest by using 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 2'-azino-bis (3-ethylbenzo thiazoline-6-sulfonic acid) (ABTS) and iron ion reducing analysis. The EAF had the highest flavonoids contents. Four fractions A, B, C and D were enrichment from EAF by polyamide resin. Fraction B had the highest flavonoids content. Thirteen chemical components of flavonoids in fraction B were first identified by Ultimate 3000 Nano LC System coupled to a Q Exactive HF benchtop Orbitrap mass spectrometer (UHPLC-HRMS/MS). Among of the thirteen chemical components, isoquercetin and daidzein were identified accurately by comparing with standard samples. Radical scavenging analysis showed that isoquercetin and EAF had strong activities. Therefore, sunflower receptacles can be used as a source of natural flavonoids. TFSR as a natural radical scavenger has potential applications in pharmaceutical industry.

2.
Sci Rep ; 11(1): 823, 2021 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-33436913

RESUMO

The challenge of decoding information about complex diseases hidden in huge number of single nucleotide polymorphism (SNP) genotypes is undertaken based on five dbGaP studies. Current genome-wide association studies have successfully identified many high-risk SNPs associated with diseases, but precise diagnostic models for complex diseases by these or more other SNP genotypes are still unavailable in the literature. We report that lung cancer, breast cancer and prostate cancer as the first three top cancers worldwide can be predicted precisely via 240-370 SNPs with accuracy up to 99% according to leave-one-out and 10-fold cross-validation. Our findings (1) confirm an early guess of Dr. Mitchell H. Gail that about 300 SNPs are needed to improve risk forecasts for breast cancer, (2) reveal an incredible fact that SNP genotypes may contain almost all information that one wants to know, and (3) show a hopeful possibility that complex diseases can be precisely diagnosed by means of SNP genotypes without using phenotypical features. In short words, information hidden in SNP genotypes can be extracted in efficient ways to make precise diagnoses for complex diseases.

3.
Sci Rep ; 10(1): 20269, 2020 11 20.
Artigo em Inglês | MEDLINE | ID: mdl-33219255

RESUMO

This paper investigates the influence of a known cue on the oblique effect in orientation identification and explains how subjects integrate cue information to identify target orientations. We design the psychophysical task in which subjects estimate target orientations in the presence of a known oriented reference line. For comparison the control experiments without the reference are conducted. Under Bayesian inference framework, a cue integration model is proposed to explain the perceptual improvement in the presence of the reference. The maximum likelihood estimates of the parameters of our model are obtained. In the presence of the reference, the variability and biases of identification are significantly reduced and the oblique effect of orientation identification is obviously weakened. Moreover, the identification of orientation in the vicinity of the reference line is consistently biased away from the reference line (i.e., reference repulsion). Comparing the predictions of the model with the experimental results, the Bayesian Least Squares estimator under the Variable-Precision encoding (BLS_VP) provides a better description of the experimental outcomes and captures the trade-off relationship of bias and precision of identification. Our results provide a useful step toward a better understanding of human visual perception in context of the known cues.

4.
Molecules ; 25(22)2020 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-33187052

RESUMO

Sunflower (Helianthus annuus L.) contains active ingredients, such as flavonoids, alkaloids and tannins. Nevertheless, few studies have focused on essential oil from the receptacle of sunflower (SEO). In this work, we investigated the chemical composition and antimicrobial and antioxidant activities of SEO. The yield of SEO was about 0.42% (v/w) by hydrodistillation. A total of 68 volatile components of SEO were putatively identified by gas chromatography-mass spectrometry (GC-MS). The main constituents of SEO were α-pinene (26.00%), verbenone (7.40%), terpinolene (1.69%) and α-terpineol (1.27%). The minimum inhibitory concentration (MIC) of SEO against P. aeruginosa and S. aureus was 0.2 mg/mL. The MIC of SEO against S. cerevisiae was 3.2 mg/mL. The MIC of SEO against E. coli and Candida albicans was 6.4 mg/mL. The results showed that SEO had high antibacterial and antifungal activities. Three different analytical assays (DPPH, ABTS and iron ion reducing ability) were used to determine the antioxidant activities. The results showed that SEO had antioxidant activities. To summarize, the results in this study demonstrate the possibility for the development and application of SEO in potential natural preservatives and medicines due to its excellent antimicrobial and antioxidant activities.

5.
ACS Omega ; 5(35): 21979-21987, 2020 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-32923756

RESUMO

Porcine deltacoronavirus (PDCoV) is a newly emerging porcine pathogenic enteric coronavirus that can cause diarrhea, vomiting, dehydration, and a high mortality rate in piglets. At present, the understanding of PDCoV pathogenesis is very limited, which seriously hinders effective prevention and control. In this study, liquid chromatography tandem-mass spectrometry (LC-MS/MS) combined with tandem mass tag (TMT) labeling was performed to compare the differential expression of proteins in PDCoV-infected and mock-infected LLC-PK cells at 18 h post-infection (hpi). In addition, the parallel reaction monitoring (PRM) technique was used to verify the quantitative proteome data. A total of 4624 differentially expressed proteins (DEPs) were quantitated, of which 128 were significantly upregulated, and 147 were significantly downregulated. Bioinformatics analysis revealed that these DEPs were involved mainly in the defense response, apoptosis, and the immune system, and several DEPs may be related to interferon-stimulated genes and the immune system. Based on DEP bioinformatics analysis, we propose that PDCoV infection may utilize the apoptosis pathway of host cells to achieve maximum viral replication. Meanwhile, the host may be able to stimulate the transcription of interferon-stimulated genes (ISGs) through the JAK/STAT signaling pathway to resist the virus. Overall, in this study, we presented the first application of proteomics analysis to determine the protein profile of PDCoV-infected cells, which provides valuable information with respect to better understanding the host response to PDCoV infection and the specific pathogenesis of PDCoV infection.

6.
Sci Total Environ ; 739: 139861, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32544678

RESUMO

We investigated the interaction between soil water supply and atmospheric evaporative demand for driving the seasonal pattern of transpiration in sky-island high-elevation forest ecosystems. Sap flow measurements were collected at 10-minute intervals for five consecutive years (2013-2017) on two co-occurring subalpine conifers, i.e. limber pine (Pinus flexilis) and bristlecone pine (Pinus longaeva). Our study site is part of the Nevada Climate-ecohydrological Assessment Network (NevCAN), and is located at 3355 m a.s.l. within an undisturbed mixed-conifer stand. We found that seasonal changes in soil moisture regulated transpiration sensitivity to atmospheric conditions. Sap flow density was mainly limited by evaporative demands under non-water limiting conditions, but was influenced only by soil moisture when water availability decreased. Daily sap flow density increased with radiation and soil moisture in June and July when soil moisture was generally above 10%, but correlated only with soil moisture in August and September when soil drought occurred. Sap flow sensitivity to vapor pressure deficit and solar radiation was therefore reduced under decreasing soil moisture conditions. Transpiration peaked in mid-to-late June during both dry and wet years, with a lower peak in late summer during wet years. Normalized mean daily canopy conductance of both species declined with decreasing soil moisture (i.e., increasing soil drought). Severe soil drying (i.e., soil moisture <7% at 20 cm depth), which was rarely detected in wet summers (2013-2014) but occurred more frequently in dry summers (2015-2017), induced a minimum in crown conductance with unchanged low-level sap flow, which might potentially trigger hydraulic failure. The minimum sap flow level under severe soil drought was higher for limber pine than bristlecone pine, possibly because of wider tracheids in limber compared to bristlecone pine. Our findings provide insights into physiological mechanisms of drought-induced stress for iconic sky-island five-needle pines located at high elevation in xeric environments.


Assuntos
Pinus , Ecossistema , Ilhas , Nevada , Transpiração Vegetal , Solo , Árvores , Estados Unidos , Água
7.
BMC Vet Res ; 16(1): 130, 2020 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-32381014

RESUMO

BACKGROUND: Porcine Deltacoronavirus (PDCoV) is a newly emerging Coronavirus that was first identified in 2012 in Hong Kong, China. Since then, PDCoV has subsequently been reported worldwide, causing a high number of neonatal piglet deaths and significant economic losses to the swine industry. Therefore, it is necessary to establish a highly sensitive and specific method for the rapid diagnosis of PDCoV. RESULTS: In the present study, a highly sensitive and specific diagnostic method using recombinase polymerase amplification combined with a lateral flow dipstick (LFD-RPA) was developed for rapid and visual detection of PDCoV. The system can be performed under a broad range of temperature conditions from 10 to 37 °C, and the detection of PDCoV can be completed in 10 min at 37 °C. The sensitivity of this assay was 10 times higher than that of conventional PCR with a lower detection limit of 1 × 102 copies/µl of PDCoV. Meanwhile, the LFD-RPA assay specifically amplified PDCoV, while there was no cross-amplification with other swine-associated viruses, including Porcine epidemic diarrhea virus (PEDV), Transmissible gastroenteritis virus (TGEV), Porcine kobuvirus (PKoV), Foot and mouth disease virus (FMDV), Porcine reproductive and respiratory syndrome virus (PRRSV), Porcine circovirus type 2 (PCV2), Classical swine fever virus (CSFV) and Seneca valley virus (SVV). The repeatability of the test results indicated that this assay had good repeatability. In addition, 68 clinical samples (48 fecal swab specimens and 20 intestinal specimens) were further tested by LFD-RPA and RT-PCR assay. The positive rate of LFD-RPA clinical samples was 26.47% higher than that of conventional PCR (23.53%). CONCLUSIONS: The LFD-RPA assay successfully detected PDCoV in less than 20 min in this study, providing a potentially valuable tool to improve molecular detection for PDCoV and to monitor the outbreak of PDCoV, especially in low-resource areas and laboratories.


Assuntos
Coronavirus/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Recombinases/metabolismo , Testes Sorológicos/veterinária , Animais , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e Especificidade , Testes Sorológicos/métodos , Suínos , Doenças dos Suínos/virologia
8.
Biotechnol Lett ; 42(10): 1907-1917, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32385744

RESUMO

OBJECTIVE: Development of an effective mucosal vaccine to induce specific immune responses against Foot-and-mouth disease virus (FMDV). RESULTS: For this purpose, the FMDV VP1 gene (SPVP1) was optimized and synthesized based on the codon bias of Lactococcus lactis (L. lactis), and then incorporated in the plasmid pNZ8148. L. lactis NZ9000 containing the pNZ8148-SPVP1 recombinant plasmid was used as an oral delivery vehicle to induce anti-FMDV mucosal and systemic immune responses in mice. After confirmation that the SPVP1 protein was expressed successfully in the recombinant L. latic, the mice were orally challenged with NZ9000-pNZ8148, NZ9000-pNZ8148-SPVP1, phosphate-buffered saline as a mock infection group, or with inactivated vaccine as a positive group. Mice immunized with NZ9000-pNZ8148-SPVP1 produced high levels of mucosal secretory IgA (sIgA), antigen-specific serum IgG, IgA, and neutralizing antibodies, and developed stronger cell-mediated immune reactions and significant T spleen lymphocyte proliferation. Furthermore, the recombinant group generated much higher levels of IFN-γ, IL-2, IL-4, IL-5, and IL-10 than the other groups. CONCLUSIONS: Potent immune responses were successfully elicited in mice with FMDV VP1 delivered through L. lactis.

9.
Arch Virol ; 165(7): 1653-1658, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32399787

RESUMO

Although porcine deltacoronavirus (PDCoV) is a significant pandemic threat in the swine population and has caused significant economic losses, information regarding the immune response in conventionally weaned pigs infected with PDCoV is scarce. Hence, the immune response in conventionally weaned pigs infected with PDCoV was assessed after challenge and rechallenge. After the first challenge, obvious diarrhea and viral shedding developed successively in all pigs in the four inoculation dose groups from 3 to 14 days postinfection (dpi), and all pigs recovered (no clinical symptoms or viral shedding) by 21 dpi. All pigs in the four groups exhibited significantly increased PDCoV-specific IgG, IgA and virus-neutralizing (VN) antibody (Ab) titers and IFN-γ levels in the serum after the first challenge. All pigs were completely protected against rechallenge at 21 dpi. The serum levels of PDCoV-specific IgG, IgA, and VN Abs increased further after rechallenge. Notably, the IFN-γ level declined continuously after 7 dpi. In addition, the levels of PDCoV-specific IgG, IgA and VN Abs in saliva increased significantly after rechallenge and correlated well with the serum Ab titers. Furthermore, the appearance of clinical symptoms of PDCoV infection in conventionally weaned pigs was delayed with reduced inoculation doses. In summary, the data presented here offer important reference information for future PDCoV animal infection and vaccine-induced immunoprotection experiments.


Assuntos
Infecções por Coronavirus/veterinária , Coronavirus/fisiologia , Doenças dos Suínos/imunologia , Animais , Anticorpos Antivirais/imunologia , Coronavirus/genética , Coronavirus/isolamento & purificação , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Diarreia/imunologia , Diarreia/virologia , Interferon gama/imunologia , Suínos , Doenças dos Suínos/virologia , Eliminação de Partículas Virais
10.
Virus Res ; 282: 197955, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32247757

RESUMO

Porcine deltacoronavirus (PDCoV) is a novel swine enteropathogenic coronavirus that causes acute diarrhea, vomiting, dehydration and mortality in neonatal piglets, resulting in significant economic losses to the pig industry. However, there is currently little information on vaccine studies and commercially available vaccines for PDCoV. Hence, herein, a PDCoV strain, CH/XJYN/2016, was successfully isolated and serially propagated in vitro, and its biological characteristics were determined. Compared to that of previously reported and recently isolated PDCoV strains from China and the United States, the S gene of the CH/XJYN/2016 strain contains novel mutations. Infection studies revealed that CH/XJYN/2016 is pathogenic to suckling piglets and conventional weaned pigs. In addition, the median pig diarrhea dose (PDD50) of PDCoV in conventional weaned pigs was determined (2.0 log10PDD50/3 mL). Furthermore, an inactivated cell-adapted CH/XJYN/2016-based vaccine candidate was developed with different adjuvants. Compared with nonvaccinated pigs, conventional weaned pigs given the inactivated vaccine developed a potent humoral immune response and showed no clinical signs or viral shedding after challenge, indicating a potent protective effect of the vaccine against PDCoV infection. Therefore, the PDCoV vaccine developed in this study is a promising vaccine candidate that can be used for the control of PDCoV infection in pigs.


Assuntos
Infecções por Coronavirus/veterinária , Coronavirus/imunologia , Coronavirus/patogenicidade , Doenças dos Suínos/virologia , Vacinas Virais/imunologia , Animais , Linhagem Celular , Coronavirus/classificação , Coronavirus/genética , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/virologia , Genoma Viral/genética , Imunogenicidade da Vacina , Mutação , Filogenia , Inoculações Seriadas , Glicoproteína da Espícula de Coronavírus/genética , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/prevenção & controle , Vacinação/veterinária , Vacinas de Produtos Inativados/imunologia
11.
Microb Pathog ; 142: 104061, 2020 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-32061916

RESUMO

To compare different nanoparticle-based nasal vaccines against foot-and-mouth disease (FMD), chitosan (CS)-coated poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) (CS/PLGA-NPs) and amino-functionalized mesoporous silica nanoparticles (Am/MSNs) loaded with FMDV recombinant plasmid (pP12A3C/IFN-CS/PLGA-NPs and pP12A3C/IFN-Am/MS-NPs) were used to induce mucosal and systemic immune responses in guinea pigs via intranasal delivery. Simultaneously, CpG oligodeoxy nucleotides (ODNs) as a vaccine adjuvant were encapsulated in chitosan-coated poly (lactic-co-glycolic acid) nanoparticles (CpG-CS/PLGA-NPs). The pP12A3C/IFN-CS/PLGA-NPs and CpG-CS/PLGA-NPs generated displayed good morphology, high stability, mean diameters of 500 and 400 nm and encapsulation efficiencies of 83.8% and 88.4%, respectively. Data from the in vitro release assay showed that plasmid and CpG were sustainably released from nanoparticles (up to 66.73% and 64%, respectively, of the total amount loaded). Guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs + CpG-CS/PLGA-NPs showed markedly higher mucosal, cellular and humoral immune responses than those administered pP12A3C/IFN-CS/PLGA-NPs or naked plasmid vaccine alone. FMDV-specific secretory immunoglobulin A (sIgA) antibodies in nasal washes were initially detected at 3 days post-vaccination with CS/PLGA-NPs loaded with plasmid. Guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs also displayed higher cellular and humoral immune responses than pP12A3C-CS/PLGA-NPs and naked plasmid vaccine alone. FMDV-specific immunoglobulin G (IgG) antibodies in serum were initially detected at 5 days post-vaccination (intramuscularly) with the naked plasmid. Finally, challenge experiments 42 days post-vaccine revealed 100% protection in guinea pigs immunized with pP12A3C/IFN-CS/PLGA-NPs + CpG-CS/PLGA-NPs and pP12A3C/IFN-CS/PLGA-NPs. However, plasmid DNA was burst released from pP12A3C/IFN-Am/MS-NPs. Our attempts to use pP12A3C/IFN-Am/MS-NPs to immunize guinea pigs failed to induce immune responses. In conclusion, CpG and IFN-α adjuvant based FMD vaccines elicit protection in guinea pigs. Moreover, CS-coated PLGA NPs present an efficient and safe mucosal immune delivery system for FMDV DNA vaccine. Data from the current study provide a foundation for understanding and further evaluating protective immune responses in pigs.

12.
Analyst ; 145(7): 2725-2730, 2020 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-32077869

RESUMO

AlGaN/GaN high electron mobility transistor (HEMT) biosensors have attracted attention due to their high sensitivity, stability, and fast response characteristics. Some related studies have been explored but a Debye screening problem exists in physiological solutions hindering the detection of bio-macromolecules. Herein, a novel fast analytical platform for electronic enzyme-linked immunosorbent assay (e-ELISA) is proposed based on AlGaN/GaN HEMT with magnetic beads (MBs); MB-based e-ELISA decouples the modified area from the sensing surface to simplify the assay. Combining the advantages of e-ELISA and MBs, the resulting analytical platform presents a sensing capability beyond the Debye-screening limit and a novel ability to be reused. This platform offers a fast response toward prostate specific antigen (PSA) and the lowest concentration of detection is 1 fg mL-1. Compared with conventional AlGaN/GaN HEMT biosensors, it shows higher sensitivity (3.73 µA dec-1) in a linear range (1 fg mL-1 to 1 pg mL-1), which is within the constraints of emergency care applications. The platform's high sensitivity and fast repeatability endow it with great potential for early and rapid diagnosis.


Assuntos
Compostos de Alumínio/química , Técnicas Biossensoriais/métodos , Ensaio de Imunoadsorção Enzimática , Gálio/química , Antígeno Prostático Específico/análise , Transistores Eletrônicos , Elétrons , Humanos , Separação Imunomagnética , Limite de Detecção , Masculino , Antígeno Prostático Específico/isolamento & purificação , Reprodutibilidade dos Testes
13.
BMC Genomics ; 21(1): 53, 2020 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-31948407

RESUMO

BACKGROUND: Ca2+ played as a ubiquitous secondary messenger involved in plant growth, development, and responses to various environmental stimuli. Calcium-dependent protein kinases (CDPK) were important Ca2+ sensors, which could directly translate Ca2+ signals into downstream phosphorylation signals. Considering the importance of CDPKs as Ca2+ effectors for regulation of plant stress tolerance and few studies on Brachypodium distachyon were available, it was of interest for us to isolate CDPKs from B. distachyon. RESULTS: A systemic analysis of 30 CDPK family genes in B. distachyon was performed. Results showed that all BdCDPK family members contained conserved catalytic Ser/Thr protein kinase domain, autoinhibitory domain, and EF-hand domain, and a variable N-terminal domain, could be divided into four subgroup (I-IV), based upon sequence homology. Most BdCDPKs had four EF-hands, in which EF2 and EF4 revealed high variability and strong divergence from EF-hand in AtCDPKs. Synteny results indicated that large number of syntenic relationship events existed between rice and B. distachyon, implying their high conservation. Expression profiles indicated that most of BdCDPK genes were involved in phytohormones signal transduction pathways and regulated physiological process in responding to multiple environmental stresses. Moreover, the co-expression network implied that BdCDPKs might be both the activator and the repressor involved in WRKY transcription factors or MAPK cascade genes mediated stress response processes, base on their complex regulatory network. CONCLUSIONS: BdCDPKs might play multiple function in WRKY or MAPK mediated abiotic stresses response and phytohormone signaling transduction in B. distachyon. Our genomics analysis of BdCDPKs could provide fundamental information for further investigation the functions of CDPKs in integrating Ca2+ signalling pathways in response to environments stresses in B. distachyon.


Assuntos
Brachypodium/enzimologia , Proteínas de Plantas/genética , Proteínas Quinases/genética , Brachypodium/genética , Perfilação da Expressão Gênica , Genoma de Planta , Sistema de Sinalização das MAP Quinases/genética , Família Multigênica , Filogenia , Proteínas de Plantas/metabolismo , Proteínas Quinases/química , Proteínas Quinases/metabolismo , Sintenia , Fatores de Transcrição/metabolismo
14.
Phys Chem Chem Phys ; 22(2): 914-918, 2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31845940

RESUMO

As a representative in-plane anisotropic two-dimensional (2D) material, germanium monoselenide (GeSe) has attracted considerable attention recently due to its various in-plane anisotropic material properties originating from the low symmetry of a puckered honeycomb structure. Although there have been plenty of reports on the in-plane anisotropic vibrational, electrical and optical properties of GeSe, the strain effect on those appealing anisotropies is still under exploration. Here we report a systematic first-principles computational investigation of strain-engineering of the anisotropic electronic properties of GeSe monolayers. We found that the anisotropic ratio of the effective mass and mobility of charge carriers (electrons and holes) of GeSe along two principle axes can be controlled by using simple strain conditions. Notably, the preferred conducting direction of GeSe can be even rotated by 90° under an appropriate uniaxial strain (>5%). Such effective strain modulation of the electronic anisotropy of GeSe monolayers provides them abundant opportunities for future mechanical-electronic devices.

15.
Biomed Res Int ; 2019: 8530273, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31687402

RESUMO

In recent years, many studies have shown that recombinant adenovirus live vector-based vaccines are a promising novel vaccine candidate against virus infection. Therefore, in this study, a new type of recombinant adenovirus expressing the spike (S) protein of porcine epidemic diarrhea virus (PEDV), rAd-PEDV-S, was generated, and its characteristics were determined. Then, its efficacy as a vaccine candidate was evaluated in 4-week-old pigs. The results showed that the S protein could be well expressed at a high level in rAd-PEDV-S-infected cells and that the viral titers could reach 1011 PFU/mL. Further animal experimental results showed that rAd-PEDV-S elicited a significant PEDV-specific humoral immune response after vaccination (P < 0.05). In addition, rAd-PEDV-S provided partial protection for pigs against the highly virulent PEDV challenge. The results presented in this study indicate that the adenovirus vector can be used as a vaccine delivery vector for the development of a PEDV vaccine and is a promising novel vaccine candidate for future prevention and control of porcine epidemic diarrhea (PED), but its efficacy still needs to be improved in the future.


Assuntos
Adenoviridae/metabolismo , Infecções por Coronavirus/virologia , Vírus da Diarreia Epidêmica Suína/metabolismo , Glicoproteína da Espícula de Coronavírus/metabolismo , Doenças dos Suínos/virologia , Adenoviridae/imunologia , Infecções por Adenoviridae/imunologia , Infecções por Adenoviridae/virologia , Animais , Anticorpos Antivirais/imunologia , Linhagem Celular , Chlorocebus aethiops , Infecções por Coronavirus/imunologia , Vírus da Diarreia Epidêmica Suína/imunologia , Suínos , Doenças dos Suínos/imunologia , Vacinas Sintéticas/imunologia , Células Vero , Vacinas Virais/imunologia
16.
Int J Mol Sci ; 20(17)2019 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-31450734

RESUMO

GT factors play critical roles in plant growth and development and in response to various environmental stimuli. Considering the new functions of GT factors on the regulation of plant stress tolerance and seeing as few studies on Brachypodium distachyon were available, we identified GT genes in B. distachyon, and the gene characterizations and phylogenies were systematically analyzed. Thirty-one members of BdGT genes were distributed on all five chromosomes with different densities. All the BdGTs could be divided into five subfamilies, including GT-1, GT-2, GTγ, SH4, and SIP1, based upon their sequence homology. BdGTs exhibited considerably divergent structures among each subfamily according to gene structure and conserved functional domain analysis, but the members within the same subfamily were relatively structure-conserved. Synteny results indicated that a large number of syntenic relationship events existed between rice and B. distachyon. Expression profiles indicated that the expression levels of most of BdGT genes were changed under abiotic stresses and hormone treatments. Moreover, the co-expression network exhibited a complex regulatory network between BdGTs and BdWRKYs as well as that between BdGTs and BdMAPK cascade gene. Results showed that GT factors might play multiple functions in responding to multiple environmental stresses in B. distachyon and participate in both the positive and negative regulation of WRKY- or MAPK-mediated stress response processes. The genome-wide analysis of BdGTs and the co-regulation network under multiple stresses provide valuable information for the further investigation of the functions of BdGTs in response to environment stresses.


Assuntos
Brachypodium/genética , Biologia Computacional , Perfilação da Expressão Gênica , Genômica , Reguladores de Crescimento de Planta/genética , Estresse Fisiológico/genética , Biologia Computacional/métodos , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Genes de Plantas , Genômica/métodos , Filogenia , Sintenia , Transcriptoma
17.
Viruses ; 11(8)2019 08 07.
Artigo em Inglês | MEDLINE | ID: mdl-31394868

RESUMO

Tumor suppressor protein p53 (p53) is a master transcription factor that plays key roles in cell cycle arrest, apoptosis, senescence, and metabolism, as well as regulation of innate immunity during virus infection. In order to facilitate their replication and spreading, viruses have evolved to manipulate p53 function through different strategies, with some requiring active p53 while others demand reduction/inhibition of p53 activity. However, there are no clear-cut reports about the roles of p53 during the infection of foot-and-mouth disease virus (FMDV), the causative agent of a highly contagious foot-and-mouth disease (FMD) of cloven-hoofed animals. Here we showed that p53 level was dynamically regulated during FMDV infection, being degraded at the early infection stage but recovered to the basal level at the late stage. Cells depleted of p53 showed inhibited FMDV replication and enhanced expression of the immune-related genes, whereas overexpression of p53 didn't affect the viral replication. Viral challenge assay with p53 knockout mice obtained similar results, with viral load decreased, histopathological changes alleviated, and lifespan extended in the p53 knockout mice. Together, these data demonstrate that basal level p53 is required for efficient FMDV replication by suppressing the innate immunity.


Assuntos
Resistência à Doença/imunologia , Vírus da Febre Aftosa/imunologia , Febre Aftosa/imunologia , Febre Aftosa/metabolismo , Interações Hospedeiro-Patógeno/imunologia , Proteína Supressora de Tumor p53/metabolismo , Animais , Linhagem Celular , Cricetinae , Febre Aftosa/virologia , Regulação da Expressão Gênica , Técnicas de Inativação de Genes , Imunidade Inata , Suínos , Proteína Supressora de Tumor p53/genética , Replicação Viral
18.
Virology ; 535: 162-170, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31306911

RESUMO

MicroRNAs (miRNAs) play important regulatory roles during interactions between virus pathogens and host cells, but whether and how they work in the case of foot-and-mouth disease virus (FMDV) is less understood. Based on a microarray-based miRNA profiling in the porcine kidney cell line PK-15, we identified 36 differentially expressed host miRNAs at the early stage of FMDV infection, among which miR-1307 was significantly induced. Functional characterization demonstrated that miR-1307 attenuated FMDV replication. Further experiments proved that miR-1307 specifically promoted the degradation of the viral structural protein VP3 indirectly through proteasome pathway. Moreover, innate immune signaling was activated and expression of immune responsive genes was significantly enhanced in the miR-1307-overexpressing clones. Together, our data demonstrated that miR-1307 suppresses FMDV replication by destabilizing VP3 and enhancing host immune response. Importantly, subcutaneous injection of miR-1307 agomir delayed the FMDV-induced lethality in suckling mice, exhibiting its therapeutic potential to control foot-and-mouth disease (FMD).


Assuntos
Proteínas do Capsídeo/metabolismo , Vírus da Febre Aftosa/imunologia , Febre Aftosa/imunologia , Imunidade Inata , Proteólise , Replicação Viral , Animais , Linhagem Celular , Modelos Animais de Doenças , Células Epiteliais/imunologia , Células Epiteliais/virologia , Vírus da Febre Aftosa/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Interações entre Hospedeiro e Microrganismos , Camundongos , Modelos Teóricos , Complexo de Endopeptidases do Proteassoma/metabolismo , Análise de Sobrevida , Suínos
19.
Arch Virol ; 164(5): 1287-1295, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30859476

RESUMO

Since 2010, continual outbreaks of highly virulent variants of porcine epidemic diarrhea virus (PEDV) belonging to genotype GII have led to serious economic losses for the Chinese swine industry. To better understand the biological characteristics and pathogenicity of the current prevalent Chinese PEDV field strains, in this study, a highly virulent Chinese genotype GIIa PEDV strain, CH/HBXT/2018, was isolated and serially propagated using Vero cells. Sequencing and phylogenetic analysis showed that strain CH/HBXT/2018 contained novel insertion and deletion mutations in the S gene region relative to the classical strain and belonged to the genotype GIIa, similar to other recently isolated PEDV strains from China and the United States. Pig infection studies indicated that the CH/HBXT/2018 strain was highly virulent in suckling piglets, and the median pig diarrhea dose (PDD50) was 8.63 log10PDD50/3 mL at 7 days postinfection (DPI). The results of the present study are important for future PEDV challenge studies and the development of new PEDV vaccines based on prevalent field strains for the prevention and control of PED in China.


Assuntos
Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína/genética , Vírus da Diarreia Epidêmica Suína/patogenicidade , Glicoproteína da Espícula de Coronavírus/genética , Animais , Linhagem Celular , China , Chlorocebus aethiops , Infecções por Coronavirus/virologia , Surtos de Doenças , Genótipo , Mutagênese Insercional/genética , Filogenia , Vírus da Diarreia Epidêmica Suína/classificação , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Deleção de Sequência/genética , Suínos , Doenças dos Suínos/virologia , Células Vero , Vacinas Virais/imunologia , Virulência/genética
20.
Appl Microbiol Biotechnol ; 103(8): 3367-3379, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30888465

RESUMO

Many recent studies have shown that flagellin fused to heterologous antigens can induce significantly enhanced humoral and cellular immune responses through its adjuvant activity. Therefore, in this study, two key B cell epitopes and a truncated VP1 (ΔVP1) protein from foot-and-mouth disease virus (FMDV) were expressed as flagellin fusion proteins in different patterns. Specifically, ΔVP1 and two duplicates of two key B cell epitopes (2×B1B2) were fused separately to the C-terminus of flagellin with a universal exogenous T cell epitope to construct FT (Flagellin-Truncated VP1) and FME (Flagellin-Multiple Epitopes). In addition, the D3 domain of flagellin was replaced by ΔVP1 in FME, yielding FTME (Flagellin-Truncated VP1-Multiple Epitopes). The immunogenicity and protective efficacy of the three fusion proteins as novel FMDV vaccine candidates were evaluated. The results showed that FT, FME, and FTME elicited significant FMDV-specific IgG responses at 10 µg/dose compared with the mock group (P < 0.05), with FTME producing the highest response. No significant differences in the antibody response to FTME were observed between different immunization routes or among adjuvants (ISA-206, poly(I·C), MPLA, and CpG-ODN) in mice. In addition, at 30 µg/dose, all three fusion proteins significantly induced neutralizing antibody production and upregulated the levels of some cytokines, including TNF-α, IFN-γ, and IL-12, in guinea pigs. Importantly, all three fusion proteins provided effective protective immunity against FMDV challenge in guinea pigs, though different protection rates were found. The results presented in this study indicate that the FTME fusion protein is a promising novel vaccine candidate for the future prevention and control of foot-and-mouth disease.


Assuntos
Flagelina/imunologia , Vírus da Febre Aftosa/imunologia , Febre Aftosa/prevenção & controle , Vacinação/métodos , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Antígenos Virais/genética , Antígenos Virais/imunologia , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/imunologia , Citocinas/sangue , Epitopos de Linfócito B/genética , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/genética , Epitopos de Linfócito T/imunologia , Feminino , Flagelina/genética , Vírus da Febre Aftosa/genética , Cobaias , Masculino , Camundongos Endogâmicos BALB C , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Vacinas Virais/administração & dosagem , Vacinas Virais/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA