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1.
Nat Commun ; 12(1): 6880, 2021 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-34824218

RESUMO

Nuclear magnetic resonance (NMR) spectroscopy is a powerful and popular technique for probing the molecular structures, dynamics and chemical properties. However the conventional NMR spectroscopy is bottlenecked by its low sensitivity. Dynamic nuclear polarization (DNP) boosts NMR sensitivity by orders of magnitude and resolves this limitation. In liquid-state this revolutionizing technique has been restricted to a few specific non-biological model molecules in organic solvents. Here we show that the carbon polarization in small biological molecules, including carbohydrates and amino acids, can be enhanced sizably by in situ Overhauser DNP (ODNP) in water at room temperature and at high magnetic field. An observed connection between ODNP 13C enhancement factor and paramagnetic 13C NMR shift has led to the exploration of biologically relevant heterocyclic compound indole. The QM/MM MD simulation underscores the dynamics of intermolecular hydrogen bonds as the driving force for the scalar ODNP in a long-living radical-substrate complex. Our work reconciles results obtained by DNP spectroscopy, paramagnetic NMR and computational chemistry and provides new mechanistic insights into the high-field scalar ODNP.

2.
Clin Neurol Neurosurg ; 212: 107067, 2021 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-34839153

RESUMO

BACKGROUND: Current researches on human carotid atherosclerosis (AS) plaques are focused on vulnerable plaques, and various methods have been clinically used to detect vulnerable plaques to prevent adverse events. GRP78 and CHOP, as markers in the endoplasmic reticulum stress (ERS), have a certain relationship with the stability of plaque tissue. METHODS: In this study, 150 plaque specimens were obtained from carotid endarterectomy (CEA). According to pathology, they were divided into two groups: stable plaque and vulnerable plaque. Immunohistochemistry was used to semi-quantitate and localize the target molecule. Western blot and RT-qPCR were used to detect the expression of GRP78 and CHOP in the samples. The receiver operating characteristic curve (ROC curve) judges the significance of the target molecule as a biomarker for the diagnosis of vulnerable plaques. RESULTS: The results of immunohistochemistry showed that the target molecules of GRP78 and CHOP were mainly expressed in inflammatory cells and vascular endothelial cells; Western blot and RT-qPCR techniques were used to detect the expression of GRP78 and CHOP in different pathlogical types of plaques, which respectively indicated that there were differential expressions. The expression in vulnerable plaques was significantly higher than that in stable plaques (P < 0.05). analysis with ROC, the areas under curves (AUC) of the GRP78 and CHOP data were calculated as 0.792 and 0.850, respectively and the combination showed the largest AUC of 0.870. CONCLUSION: In endoplasmic reticulum stress, GRP78 and CHOP are significantly higher expressions in vulnerable plaques than stable's, which indicated that GRP78 and CHOP played a certain role in the occurrence and development of human carotid atherosclerosis and vulnerable plaques; GRP78 and CHOP are promising molecular biomarkers for identifying the endoplasmic reticulum stress situation, atherosclerosis and plaque stability. They also could provide a potential drug targets for the prevention and treatment of atherosclerosis.

3.
Front Cardiovasc Med ; 8: 736215, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34712709

RESUMO

Background: Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a transmembrane glycoprotein that mediates uptake of oxidized low-density lipoprotein (ox-LDL) into cells. Previous studies had shown that LOX-1 deletion had a potential to inhibit cardiac fibrosis in mouse models of hypertension and myocardial infarction. Whether LOX-1 deletion also affects cardiac fibrosis associated with aging still remains unknown. The aim of this study was to investigate the effect of LOX-1 deletion on myocardial fibrosis in the aged mice. Methods: C57BL/6 mice and LOX-1 knockout (KO) mice with C57BL/6 background were studied to the age of 60 weeks. Both genotypes of aged mice were exposed to angiotensin II (Ang II) or saline for additional 4 weeks. The mice were then sacrificed, and myocardial fibrosis, reactive oxygen species (ROS) and expression of LOX-1, fibronectin, collagens, p22phox, and gp91phox were measured. Results: LOX-1 deletion markedly reduced Ang II-mediated rise of blood pressure in the aged mice (vs. saline-treated mice). LOX-1 deletion also limited fibrosis and decreased fibronectin and collagen-3 expression in the hearts of aged mice, but not the expression of collagen-1 and collagen-4. LOX-1 deletion also inhibited ROS production and p22phox expression. As the aged mice were exposed to Ang II for 4 weeks (resulting in hypertension), LOX-1 deletion more pronounced inhibiting myocardial fibrosis and ROS production, and decreasing expression of fibronectin, collagen-1, collagen-2, collagen-3, p22phox, and gp91phox. Conclusion: LOX-1 deletion limited fibrosis and ROS production in the hearts of aged mice. This effect was more pronounced in the aged mice with hypertension induced by Ang II infusion.

4.
Sci Total Environ ; : 150831, 2021 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-34627884

RESUMO

Peatlands in northeast China are experiencing severe climate warming. Most studies on peatlands focus on the responses of CH4 dynamics to temperature. However, they rarely consider the synchronous changes in the composition of plant communities caused by the expansion of vascular plants. In this study, an experiment combined warming with the manipulation of plants to examine the concentrations of CH4 porewater and its fluxes in the mesocosm. We found that warming increased the concentration of CH4 and its fluxes relative to the control treatments, and it was strongly modulated by plant richness and functional types. The average CH4 fluxes in the warming and non-warming mesocosms varied from 72.10 to 119.44 and 97.95 to 194.43 mg m-2 h-1, respectively. Plant species richness significantly increased CH4 flux at the warming level of 3.2 °C (P < 0.01). The presence of vascular plants, such as Carex globularis and Vaccinium uliginosum, significantly increased the CH4 fluxes after warming had occurred. Our results suggest that the distinct response of CH4 to richness and species primarily stemmed from the direct or indirect effects of plant biomass and functional characteristics. Therefore, more consideration should be given to the diversity changes caused by vascular plant expansion when estimating CH4 flux in boreal peatland, especially in the context of future climate warming.

5.
Cell Rep ; 36(13): 109761, 2021 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-34592151

RESUMO

Drosophila Vago is a small antiviral peptide. Its ortholog in Culex mosquito was found to be an interferon-like cytokine that limits virus replication through activating Jak/Stat signaling. However, this activation is independent of Domeless, the sole homolog of vertebrate type I cytokine receptor. How Vago activates the Jak/Stat pathway remains unknown. Herein, we report this process is dependent on integrin in kuruma shrimp (Marsupenaeus japonicus). Shrimp Vago-like (MjVago-L) plays an antiviral role by activating the Jak/Stat pathway and inducing Stat-regulated Ficolin. Blocking integrin abrogates the role of MjVago-L. The interaction between MjVago-L and integrin ß3 is confirmed. An Asp residue in MjVago-L is found critical for the interaction and MjVago-L's antiviral role. Moreover, Fak, a key adaptor of integrin signaling, mediates MjVago-L-induced Jak/Stat activation. Therefore, this study reveals that integrin, as the receptor of MjVago-L, mediates Jak/Stat activation. The establishment of the MjVago-L/integrin/Fak/Jak/Stat/Ficolin axis provides insights into antiviral cytokine signaling in invertebrates.

6.
Phys Chem Chem Phys ; 23(37): 21001-21012, 2021 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-34522933

RESUMO

A fixed multi-site interaction charge (FMIC) model was proposed for the accurate prediction of intermolecular electrostatic interactions based on the quantum mechanical linear response of a molecule to an external electric field. In such a model, some additional off-center interaction sites were added for capturing multipole interactions for a given molecule. By multivariate least-square fitting analysis of the calculated QM/MM interactions of a given molecule with the electrostatic environment and the electrostatic potentials of the environment at the pre-defined distributed interaction sites, the FMIC of the molecule was obtained. The model system of CO in myoglobin (Mb) was utilized to demonstrate the derivation of the FMIC. The accuracy of FMIC in predicting the electrostatic interactions between CO and the Mb environment was investigated using 10 000 different Mb-CO configurations generated from the 400 ps QM/MM MD simulation. In comparison to the QM/MM calculations at the B3LYP/aug-cc-pVTZ/ff99SB level, the mean unsigned error (MUE) of the results based on the FMIC model was merely 0.10 kcal mol-1, and the root mean square error (RMSE) was only 0.13 kcal mol-1, which are significantly lower than the results predicted by the ESP charge model (MUE = 1.45 kcal mol-1, and RMSE = 1.7 kcal mol-1, respectively). The transferability of FMIC was tested by applying the obtained FMIC in the wild type Mb-CO system to the mutants of V68F and H64L Mb-CO systems. The MUEs of the obtained results for 10 000 different configurations are both smaller than 0.2 kcal mol-1 for the V68F and H64L Mb-CO systems in comparison to the B3LYP/aug-cc-pVTZ/ff99SB calculations, and the RMSEs are also lower than 0.2 kcal mol-1 for both mutants. The applications of FMIC were extended to model the electrostatic interactions between a hydrogen fluoride molecule and 492 waters in a truncated octahedron box; our study showed that the FMIC could give satisfactory results with a MUE of 0.12 kcal mol-1 and a RMSE of 0.16 kcal mol-1 in comparison to the B3LYP/aug-cc-pVDZ/TIP3P calculations for 10 000 different configurations generated using the 10 ns classical MD simulation. Therefore, the FMIC method provides an accurate and efficient tool for predicting intermolecular electrostatic interactions, which can be utilized in the future development of molecular force fields.

7.
Front Pharmacol ; 12: 699949, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34512335

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or COVID-19 infection is the cause of the ongoing global pandemic. Mortality from COVID-19 infection is particularly high in patients with cardiovascular diseases. In addition, COVID-19 patients with preexisting cardiovascular comorbidities have a higher risk of death. Main cardiovascular complications of COVID-19 are myocardial infarction, myocarditis, acute myocardial injury, arrhythmias, heart failure, stroke, and venous thromboembolism. Therapeutic interventions in terms of drugs for COVID-19 have many cardiac adverse effects. Here, we review the relative therapeutic efficacy and adverse effects of anti-COVID-19 drugs.

8.
Cerebrovasc Dis ; : 1-9, 2021 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-34407532

RESUMO

BACKGROUND AND PURPOSE: Existing studies on the association between hemoglobin values and stroke outcomes mostly focus on the lower side and mortality, often the only and primary endpoint. The current study was conducted to assess the association between hemoglobin concentration and a variety of poor stroke outcomes in patients with acute ischemic stroke. METHODS: We studied 8,321 patients enrolled in the China National Stroke Registry (CNSR) between 2007 and 2008. Patients were divided into 7 groups, and a logistic regression model was used to evaluate the association. Endpoints of interest included 1-year all-cause mortality, stroke recurrence, combined endpoint, and stroke disability. Stroke disability was defined as a modified Rankin Scale of 2-6. RESULTS: Patients with low and high hemoglobin values (≤11.6 g/dL and >16.1 g/dL) had higher proportion of poststroke adverse events than those in other groups. As compared with the fourth group of hemoglobin values of 13.5-14.2 g/dL, the adjusted odds ratios (ORs) with 95% confidence interval (CI) of low hemoglobin values (≤11.6 g/dL) were 2.25 (1.72-2.93) for all-cause mortality, 1.30 (1.04-1.61) for stroke recurrence, 1.63 (1.33-2.01) for combined endpoint, and 1.37 (1.12-1.67) for stroke disability, respectively. And, the ORs of high hemoglobin values (>16.1 g/dL) for adverse stroke outcomes were 1.72 (1.25-2.37), 1.43 (1.13-1.82), 1.43 (1.13-1.81), and 1.31 (1.06-1.63), respectively. Stratified analysis showed significant interactions between sex and categories of hemoglobin values for all-cause mortality (p = 0.05), stroke recurrence (p = 0.03), and combined endpoint (p = 0.01) but not for stroke disability (p = 0.24). CONCLUSIONS: Our study found both low and high hemoglobin values were associated with adverse stroke outcomes including all-cause mortality, stroke recurrence, combined endpoint, and stroke disability, which showed a U-shaped association. And, significant interactions between sex and hemoglobin concentration on all-cause mortality and stroke recurrence were also identified.

9.
Am J Physiol Regul Integr Comp Physiol ; 321(5): R639-R654, 2021 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-34431382

RESUMO

After an ischemic event, there is activation of fibroblasts leading to scar formation. It is critical to limit the profibrotic remodeling and activate the reparative remodeling phase to limit cardiac diastolic dysfunction. Mesenchymal stem cell (MSC) exosomes offer significant protection against ischemia-related systolic dysfunction. Here, we studied if MSC exosomes would offer protection against profibrotic events in mouse hearts subjected to acute ischemia [1 h left coronary artery (LCA) occlusion] or chronic ischemia (7 days LCA occlusion). After acute ischemia, there was activation of inflammatory signals, more in the peri-infarct than in the infarct area, in the saline (vehicle)-treated mice. At the same time, there was expression of cardiac remodeling signals (vimentin, collagens-1 and -3, and fibronectin), more in the infarct area. Treatment with MSC exosomes before LCA ligation suppressed inflammatory signals during acute and chronic ischemia. Furthermore, exosome treatment promoted pro-reparative cardiac extracellular matrix (ECM) remodeling in both infarct and peri-infarct areas by suppressing fibronectin secretion and by modulating collagen secretion to reduce fibrotic scar formation through altered cellular signaling pathways. Proteomics study revealed intense expression of IL-1ß and activation of profibrotic signals in the saline-treated hearts and their suppression in MSC exosome-treated hearts. To our knowledge, this is the first report on the infarct and peri-infarct area proteomics of ischemic mice hearts to explain MSC exosome-mediated suppression of scar formation in the ischemic mouse hearts.


Assuntos
Exossomos/transplante , Fibroblastos/metabolismo , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Isquemia Miocárdica/cirurgia , Miocárdio/metabolismo , Proteoma , Proteômica , Remodelação Ventricular , Animais , Western Blotting , Linhagem Celular , Movimento Celular , Modelos Animais de Doenças , Eletroforese em Gel de Poliacrilamida , Exossomos/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Fibroblastos/patologia , Fibrose , Humanos , Mediadores da Inflamação/metabolismo , Interleucina-1beta/metabolismo , Masculino , Espectrometria de Massas , Células-Tronco Mesenquimais/metabolismo , Camundongos Endogâmicos C57BL , Isquemia Miocárdica/metabolismo , Isquemia Miocárdica/patologia , Miocárdio/patologia
10.
J Stroke Cerebrovasc Dis ; 30(10): 106045, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34418671

RESUMO

BACKGROUND: Matrix metalloproteinase 10 (MMP-10) has a close relationship with carotid atherosclerosis (CAS) and cerebral infarction. The MMP-10 rs17435959 polymorphism causes a leucine to valine transition at codon 4 in exon 1 of the MMP-10 gene and may have functional effects. OBJECTIVES: To investigate the relationship between the MMP-10 rs17435959 polymorphism and the formation and stability of CAS plaques. MATERIALS AND METHODS: The present case-control study contains 738 visitors who came to our health examination center for the first time. According to the carotid ultrasound examinations, visitors were classified into the vulnerable plaque group (41-86 years old, 141 male, 105 female), the stable plaque group (41-86 years old, 141 male, 105 female) and the no plaque group (41-85 years old, 141 male, 105 female). All visitors in the three groups were sex- and- age-matched, and cardiovascular and cerebrovascular diseases were absent. The polymorphism was genotyped by real-time polymerase chain reaction- restriction. RESULTS: Compared to the GG genotype, the frequency of the CC and CG genotypes was significantly more common in the vulnerable plaque group than in the no plaque group (18.7% vs. 7.7%, unadjusted P = 0.002). Moreover, compared to the G allele, the frequency of the C allele was significantly more common in the vulnerable plaque group than in the no plaque group (10.4% vs. 3.9%, unadjusted P = 0.000) and in the vulnerable plaque group than in the stable plaque group (10.4% vs. 5.1%, unadjusted P = 0.008). Binary logistic regression showed that the CC and CG genotype was independent risk factor for the formation (P = 0.019, OR = 1.961, 95% CI [1.117, 3.444]) and vulnerability (P = 0.035, OR = 1.842, 95% CI [1.045, 3.247]) of CAS plaques. Moreover, individuals who have the C allele showed a higher level of fibrinogen, which was an independent risk factor for the formation of CAS plaques (P = 0.000, OR = 2.425, 95% CI [1.475, 3.985]). CONCLUSIONS: The rs17435959 polymorphism was associated with the formation and vulnerability of CAS plaques. Individuals who had variant-type MMP-10 showed higher levels of fibrinogen, which promoted the formation of CAS plaques.


Assuntos
Doenças das Artérias Carótidas/genética , Metaloproteinase 10 da Matriz/genética , Placa Aterosclerótica , Polimorfismo de Nucleotídeo Único , Adulto , Idoso , Idoso de 80 Anos ou mais , Doenças das Artérias Carótidas/diagnóstico por imagem , Doenças das Artérias Carótidas/enzimologia , Estudos de Casos e Controles , Feminino , Fibrinogênio/análise , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Fenótipo , Prognóstico , Medição de Risco , Fatores de Risco , Ruptura Espontânea
11.
J Virol ; 95(19): e0092321, 2021 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-34260286

RESUMO

Peroxiredoxin 1 (PRDX1) is a cellular antioxidant enzyme that is crucial for diverse fundamental biological processes, such as autophagy, inflammation, and carcinogenesis. However, molecular mechanisms underpinning its diverse roles are not well understood. Here, we report that PRDX1 positively regulates interferon (IFN) induction and that pseudorabies virus (PRV) targets PRDX1 to evade IFN induction. PRV UL13 encodes a serine/threonine kinase important for PRV infection, although its biological function remains obscure. We identified PRDX1 as a UL13-interacting protein. Virological and biochemical assays demonstrate that PRDX1 promotes IFN induction by interacting with TANK-binding kinase 1 (TBK1) and IκB kinase ε (IKKε). Conversely, UL13 accelerates PRDX1 degradation via the ubiquitin-proteosome pathway in a kinase-dependent manner. In doing so, PRV inhibits IFN induction during productive infection, which requires PRDX1 expression. This study uncovers an essential role of PRDX1 in the innate immune response and reveals a new viral immune evasion strategy to counteract cellular defenses. IMPORTANCE PRV interacts with numerous cellular proteins during productive infection. Here, we demonstrated the interaction of viral protein UL13 with the antioxidant enzyme PRDX1, which functions in multiple signal transduction pathways. We found that PRDX1 participates in the type I IFN pathway by interacting with TBK1 and IKKε, thereby negatively regulating PRV propagation. However, UL13 ubiquitinates PRDX1, which routes PRDX1 into proteasomes for degradation and effectively reduces its expression. These results illuminate the fundamental role that PRDX1 plays in the IFN pathway, and they identify a potential target for the control of PRV infection.


Assuntos
Herpesvirus Suídeo 1/fisiologia , Quinase I-kappa B/metabolismo , Imunidade Inata , Peroxirredoxinas/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Virais/metabolismo , Animais , Linhagem Celular , Células HEK293 , Herpesvirus Suídeo 1/imunologia , Humanos , Evasão da Resposta Imune , Interferon Tipo I/biossíntese , Mutação , Complexo de Endopeptidases do Proteassoma/metabolismo , Transdução de Sinais , Ubiquitinação , Proteínas Virais/genética , Replicação Viral
12.
J Virol ; 95(18): e0021021, 2021 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-34160254

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important endemic swine pathogens, causing enormous losses in the global swine industry. Commercially available vaccines only partially prevent or counteract the virus infection and correlated losses. PRRSV's replication mechanism has not been well understood. In this study, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was screened to bind with the viral major envelope glycoprotein 5 (GP5) after PRRSV infection. The interacting sites are located within a 13-amino-acid (aa) region (aa 93 to 105) of GP5 and at Lys227 of GAPDH. Interestingly, viral GP5 restricts the translocation of GAPDH from the cytoplasm to the nucleus. Moreover, cytoplasmic GAPDH facilitates PRRSV replication by virtue of its glycolytic activity. The results suggest that PRRSV GP5 restricts GAPDH to the nucleus and exploits its glycolytic activity to stimulate virus replication. The data provide insight into the role of GAPDH in PRRSV replication and reveal a potential target for controlling viral infection. IMPORTANCE PRRSV poses a severe economic threat to the pig industry. PRRSV GP5, the major viral envelope protein, plays an important role in viral infection, pathogenicity, and immunity. However, interactions between GP5 and host proteins have not yet been well studied. Here, we show that GAPDH interacts with GP5 through binding a 13-aa sequence (aa 93 to 105) in GP5, while GP5 interacts with GAPDH at the K277 amino acid residue of GAPDH. We demonstrate that GP5 interacts with GAPDH in the cytoplasm during PPRSV infection, inhibiting GAPDH entry into the nucleus. PRRSV exploits the glycolytic activity of GAPDH to promote viral replication. These results enrich our understanding of PRRSV infection and pathogenesis and open a new avenue for antiviral prevention and PRRSV treatment strategies.


Assuntos
Núcleo Celular/metabolismo , Citoplasma/metabolismo , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Proteínas do Envelope Viral/metabolismo , Replicação Viral , Animais , Gliceraldeído-3-Fosfato Desidrogenases/genética , Células HEK293 , Humanos , Síndrome Respiratória e Reprodutiva Suína/genética , Síndrome Respiratória e Reprodutiva Suína/metabolismo , Suínos , Proteínas do Envelope Viral/genética
13.
Vet Microbiol ; 257: 109076, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33957572

RESUMO

The Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-borne hemorrhagic zoonotic disease, which is potentially fatal in human with mortality rates ranging from 16.2%-32%. The rabies virus (RABV) LBNSE vector expressing foreign antigens have shown considerable promise as vaccines against viral diseases, which is effective and safe. In the present study, we generated a recombinant RABV rLBNSE-Gn expressing a SFTSV glycoprotein Gn by reverse genetic technology to control rabies and SFTS in animals. An extra insertion of Gn gene did not impact replication of the recombinant virus rLBNSE-Gn in NA and BHK-21 cells compared to the parent rLBNSE strain. The SFTSV Gn gene together with RABV N and G genes were efficiently expressed in rLBNSE-infected Vero cells by immunostaining and immune blots. A single dose of 107 FFU of the rLBNSE-Gn intramuscularly inoculated in BALB/c mice induced rapid and robust humoral responses against both RABV and SFTSV without any signs of disease or weight loss. Compared to the rLBNSE and DMEM groups, the extra Gn expression contributed to the recruitments and/or activations of the dendritic cells and B cells from inguinal lymph nodes of BALB/c mice vaccinated with rLBNSE-Gn. The protective efficacy of rLBNSE-Gn against SFTSV in C57BL/6 mice was evaluated, and the virus loading in the spleens reduced to 10 TCID50/mg at 7 days post SFTSV infections, which indicated that the rLBNSE-Gn conferred efficacious protective immune responses from SFTSV in C57BL/6 mice. All the mice immunization with rLBNSE-Gn and rLBNSE survived after a lethal RABV challenge, suggesting a 100 % protection from RABV. Therefore, the rLBNSE-Gn would be a promising bivalent candidate vaccine against SFTS and rabies in animals.


Assuntos
Anticorpos Antivirais/sangue , Vetores Genéticos , Phlebovirus/imunologia , Vírus da Raiva/genética , Raiva/prevenção & controle , Febre Grave com Síndrome de Trombocitopenia/prevenção & controle , Vacinas Virais/imunologia , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Phlebovirus/genética , Vacinas Antirrábicas/administração & dosagem , Vacinas Antirrábicas/imunologia , Vírus da Raiva/imunologia , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/imunologia , Vacinas Combinadas/administração & dosagem , Vacinas Combinadas/genética , Vacinas Combinadas/imunologia , Vacinas de Produtos Inativados/administração & dosagem , Vacinas de Produtos Inativados/imunologia , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/imunologia , Vacinas Virais/administração & dosagem , Vacinas Virais/genética
14.
Vet Microbiol ; 258: 109104, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34004569

RESUMO

Pseudorabies is a highly infectious disease with severe clinical symptoms, causing acute death in infected pigs and leading to substantial economic losses among swine producers. In this study, a vaccine candidate strain in which the protein kinase UL13 gene was deleted was constructed with the CRISPR/Cas9 system based on the recombinant pseudorabies virus (PRV) ZJ01-ΔgI/gE/TK. Pigs immunized with ZJ01-ΔgI/gE/TK or ZJ01-ΔgI/gE/TK/UL13 produced high levels of anti-gB antibodies and virus-neutralizing antibodies. ZJ01-ΔgI/gE/TK/UL13 provided greater protective efficacy against challenge with PRV variant strain ZJ01 than did Bartha-K61 or ZJ01-ΔgI/gE/TK. The pigs vaccinated with ZJ01-ΔgI/gE/TK/UL13 excreted significantly less virus than those vaccinated with Bartha-K61 or ZJ01-ΔgI/gE/TK. The viral loads in the lungs of pigs treated with ZJ01-ΔgI/gE/TK/UL13 were lower than those in pigs treated with ZJ01-ΔgI/gE/TK after challenge with PRV variant strain ZJ01. These data indicated that ZJ01-ΔgI/gE/TK/UL13 had greater protective efficacy and safety than the commercial ZJ01-ΔgI/gE/TK and Bartha-K61 vaccines, and could be developed as a promising vaccine candidate for the prevention and control of this disease.


Assuntos
Herpesvirus Suídeo 1/genética , Vacinas contra Pseudorraiva/imunologia , Pseudorraiva/virologia , Doenças dos Suínos/prevenção & controle , Animais , Linhagem Celular , Interferon beta/genética , Interferon beta/metabolismo , Pseudorraiva/imunologia , Testes Sorológicos , Suínos , Doenças dos Suínos/virologia
15.
Vet Microbiol ; 256: 109038, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33845332

RESUMO

Cholesterol 25-hydroxylase (CH25 H), as a host restriction factor, has been reported to take a broad-spectrum antiviral effect. However, the role of CH25H in Senecavirus A (SVA) infection remains unknown. In this study, we first demonstrate that overexpression of CH25H inhibits SVA replication. Consistently, knockdown or knockout of the endogens CH25H promotes SVA infection. Further, the anti-SVA effect of 25-hydroxycholesterol (25HC), which is the product of CH25H, operates via inhibition of viral attachment and replication. On the other hand, the CH25H mutant (CH25H-M) lacking hydroxylase activity still restricts SVA infection, which can selectively interact and degrade SVA 3A protein via the ubiquitin-proteasome manner. Altogether, these results suggest that CH25H has an antiviral function in SVA infection and provides an alternative manner to control SVA.


Assuntos
Infecções por Picornaviridae/prevenção & controle , Picornaviridae/fisiologia , Esteroide Hidroxilases/metabolismo , Replicação Viral , Animais , Antivirais , Linhagem Celular , Cricetinae , Técnicas de Silenciamento de Genes , Técnicas de Inativação de Genes , Mutação , Infecções por Picornaviridae/virologia , Esteroide Hidroxilases/genética
16.
J Chem Phys ; 154(13): 134107, 2021 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-33832260

RESUMO

The interactions of the polar chemical bonds such as C=O and N-H with an external electric field were investigated, and a linear relationship between the QM/MM interaction energies and the electric field along the chemical bond is established in the range of weak to intermediate electrical fields. The linear relationship indicates that the electrostatic interactions of a polar group with its surroundings can be described by a simple model of a dipole with constant moment under the action of an electric field. This relationship is employed to develop a general approach to generating an electrostatic energy-based charge (EEC) model for molecules containing single or multiple polar chemical bonds. Benchmark test studies of this model were carried out for (CH3)2-CO and N-methyl acetamide in explicit water, and the result shows that the EEC model gives more accurate electrostatic energies than those given by the widely used charge model based on fitting to the electrostatic potential (ESP) in direct comparison to the energies computed by the QM/MM method. The MD simulations of the electric field at the active site of ketosteroid isomerase based on EEC demonstrated that EEC gave a better representation of the electrostatic interaction in the hydrogen-bonding environment than the Amber14SB force field by comparison with experiment. The current study suggests that EEC should be better suited for molecular dynamics study of molecular systems with polar chemical bonds such as biomolecules than the widely used ESP or RESP (restrained ESP) charge models.

17.
Vet Res ; 52(1): 52, 2021 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-33766129

RESUMO

Long non-coding RNAs (lncRNAs) are a new arm of gene regulatory mechanism as discovered by sequencing techniques and follow-up functional studies. The lncRNAs regulation of pseudorabies virus (PRV) infection has rarely been reported so far. Using RNA sequencing analysis, 225 lncRNAs with significant altered expressions in 3D4/21 cells infected with PRV (ZJ01) were identified. Five lncRNAs upregulated in PRV-infected cells were verified in cells infected with different PRV strains by qRT-PCR. By down- and up-regulation of lnc641, the accelerating effect of lnc641 on PRV replication was confirmed. Furthermore, we found that lnc641 regulated PRV replication by inhibiting the JAK-STAT1 pathway. This study suggests that lnc641 could be a new host factor target for developing antiviral therapies against PRV infection.


Assuntos
Regulação Viral da Expressão Gênica , Herpesvirus Suídeo 1/fisiologia , RNA Longo não Codificante/genética , RNA Viral/genética , Replicação Viral/genética , Animais , Linhagem Celular , Herpesvirus Suídeo 1/genética , RNA Longo não Codificante/metabolismo , RNA Viral/metabolismo , Sus scrofa
18.
J Immunol ; 206(6): 1140-1150, 2021 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-33526439

RESUMO

Intestinal microbiota are closely related to host physiology. Over the long course of evolution and interaction, both commensal bacteria and their host have evolved multiple strategies to adapt to each other. However, in invertebrates, the regulatory mechanism of intestinal microbiota homeostasis is largely unknown. In the current study, a digestive tract-specific C-type lectin, designated as CTL33, was identified because of its abundance and response to bacteria in the intestine of kuruma shrimp (Marsupenaeus japonicus). Silencing of CTL33 expression led directly to intestinal dysbiosis, tissue damage, and shrimp death. CTL33 could facilitate biofilm formation by the intestinal bacteria. This function originated from its unique architecture, with a lectin domain responsible for bacteria recognition and a coiled coil region that mediated CTL33 dimerization and cross-linked the bacteria into a biofilm-like complex. By mediating the formation of a biofilm, CTL33 promoted the establishment of intestinal bacteria in intestine and maintained the homeostasis of the microbiota. Thus, to our knowledge, we demonstrated a new mechanism of C-type lectin-mediated biofilm formation by intestinal bacteria, providing new insights into intestinal homeostasis regulation in invertebrates.


Assuntos
Proteínas de Artrópodes/metabolismo , Bactérias/imunologia , Microbioma Gastrointestinal/imunologia , Lectinas Tipo C/metabolismo , Penaeidae/imunologia , Animais , Proteínas de Artrópodes/genética , Biofilmes , Disbiose/genética , Disbiose/imunologia , Disbiose/microbiologia , Técnicas de Silenciamento de Genes , Homeostase/imunologia , Interações entre Hospedeiro e Microrganismos/imunologia , Lectinas Tipo C/genética , Penaeidae/metabolismo , Penaeidae/microbiologia , Domínios Proteicos
19.
Angew Chem Int Ed Engl ; 60(14): 7860-7865, 2021 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-33393709

RESUMO

Carbonic acid, H2 CO3 , is of fundamental importance in nature both in living and non-living systems. Providing direct spectroscopic evidence for carbonic acid formation is however a challenge. Here we provide clear evidence by in situ attenuated total reflection IR spectroscopy combined with modulation excitation spectroscopy and phase-sensitive detection that CO2 adsorption on ice surfaces is accompanied by carbonic acid formation. We demonstrate that carbonic acid can be formed from CO2 on ice in the absence of high-energy irradiation and without protonation by strong acids. The formation of carbonic acid is favored at low temperature, whereas at high temperature it rapidly dissociates to form bicarbonate (HCO3 - ) and carbonate (CO3 2- ). The direct formation of carbonic acid from adsorption of CO2 on ice could play a role in the upper troposphere in cirrus clouds, where all the necessary ingredients to form carbonic acid, that is, low temperature, CO2 gas, and ice, are present.

20.
Mol Cell Biochem ; 476(4): 1691-1704, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33423165

RESUMO

Mesenchymal stem cell (MSC) exosomes may limit cardiac injury, and even reverse cardiac damage in animal models of ischemia. To understand exosome-mediated improvement in cardiac function we examined the proteomic alternations in the MSC exosome-treated mice hearts subjected to left coronary artery (LCA) ligation, with particular emphasis on peri-infarct areas. At 7 days after LCA ligation, left ventricular end systolic thickness, infarct size and survival of mice were studied. Mass spectrometric analysis of infarct and peri-infarct areas was carried out. Expression of inflammatory markers (LOX-1 and NLRP3) and cell death markers (Bax, Bcl-2, Caspases 1 and 3 and GSDMD) were investigated by Western blots and immunofluorescence. Proteomic analysis of the infarct and peri-infarct areas in saline-treated hearts revealed differentially expressed proteins involved in inflammation and apoptotic cell death, while showing depletion of processes governing cell death. Exosome treatment significantly improved the proteomic profile in both infarct and peri-infarct areas, more so in the peri-infarct areas. The infarct size was smaller (9 ± 1%), and cardiac contractile function (fractional shortening) was preserved in the exosome-treated mice (28 ± 2%). Survival of exosome-treated mice was also better. White blood cell accumulation in and around the infarct area, expression of LOX-1 and NLRP3 inflammasome, and markers of cell death (cleaved Caspase-3, Caspase-1, GSDMD, Bcl-2 and Bax) were dramatically reduced by MSC exosome treatment (all p < 0.01). In cultured primary mouse cardiomyocytes, treatment with MSC exosomes essentially reversed inflammation-induced pro-apoptotic and inflammatory signals (p < 0.01). MSC exosomes exert their cardioprotective effects by suppressing inflammation and pro-apoptotic processes, particularly in the peri-infarct areas, resulting in preservation of cardiac function after LCA ligation.


Assuntos
Exossomos , Células-Tronco Mesenquimais/metabolismo , Infarto do Miocárdio , Animais , Linhagem Celular Transformada , Exossomos/metabolismo , Exossomos/patologia , Exossomos/transplante , Humanos , Masculino , Células-Tronco Mesenquimais/patologia , Camundongos , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Infarto do Miocárdio/prevenção & controle
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