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The foot-and-mouth disease virus (FMDV) is the causative agent of FMD, a highly infectious and devastating viral disease of domestic and wild cloven-hoofed animals. FMD affects livestock and animal products' national and international trade, causing severe economic losses and social consequences. Currently, inactivated vaccines play a vital role in FMD control, but they have several limitations. The genetic code expansion technology provides powerful strategies for generating premature termination codon (PTC)-harbouring virus as a live but replication-incompetent viral vaccine. However, this technology has not been explored for the design and development of new FMD vaccines. In this study, we first expanded the genetic code of the FMDV genome via a transgenic cell line containing an orthogonal translation machinery. We demonstrated that the transgenic cells stably integrated the orthogonal pyltRNA/pylRS pair into the genome and enabled efficient, homogeneous incorporation of unnatural amino acids into target proteins in mammalian cells. Next, we constructed 129 single-PTC FMDV mutants and four dual-PTC FMDV mutants after considering the tolerance, location, and potential functions of those mutated sites. Amber stop codons individually substituted the selected amino acid codons in four viral proteins (3D, L, VP1, and VP4) of FMDV. We successfully rescued PTC-FMDV mutants, but the amber codon unexpectedly showed a highly degree of mutation rate during PTC-FMDV packaging and replication. Our findings highlight that the genetic code expansion technology for the generation of PTC-FMD vaccines needs to be further improved and that the genetic stability of amber codons during the packaging and replication of FMDV is a concern.
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Códon sem Sentido , Códon de Terminação , Vírus da Febre Aftosa/genética , Febre Aftosa/virologia , Proteínas Virais/genética , Replicação Viral , Animais , Animais Geneticamente Modificados , Cricetinae , Vírus da Febre Aftosa/isolamento & purificação , Genoma Viral , Rim/virologia , MutaçãoRESUMO
The interaction between nanoparticles and polymers has been of great interest in colloidal theory and novel materials. For example, the properties of polyacrylamide solutions and gels, which are usually used for conformance control and water shut-off in oilfields, can be improved with the addition of nanoparticles. This underlying mechanism and its applicability are investigated in this paper. A strong relationship between the phase behaviors of nanoparticles in polymer solutions and their enhancement effect on the rheology of the nanocomposite polymer solutions and gels was observed. Experiment results showed that the stability of nanoparticles was dependent on several factors, including pH, salinity, and polymer type. At neutral pH conditions, the tendency of the aggregation of nanoparticles was strengthened upon increasing the salinity, polymer concentration, and electronegativity of the polymers. Rheological measurements showed that nanoparticles could improve the viscosity of polymer solutions or the fracture stress of gels only if nanoparticles were aggregated in the corresponding systems. In addition, these rheological parameters significantly increased with increasing salinity and nanoparticle concentration. As a result, the mobility ratio of polymer solutions may be increased several times by the addition of nanoparticles. Referring to the gels, their rupture pressure gradient in the ideal model was also found to increase with nanoparticle concentration. In particular, if the nanoparticle concentration was sufficiently high (reaching 2%), the formed gels would not be destroyed by the injected water, but rather functionally act as a porous medium for permeation.
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Metabolic flux analysis was used to reveal the metabolic distributions in Gluconacetobacter xylinus (CGMCC no. 2955) cultured on different carbon sources. Compared with other sources, glucose, fructose, and glycerol could achieve much higher bacterial cellulose (BC) yields from G. xylinus (CGMCC no. 2955). The glycerol led to the highest BC production with a metabolic yield of 14.7 g/mol C, which was approximately 1.69-fold and 2.38-fold greater than that produced using fructose and glucose medium, respectively. The highest BC productivity from G. xylinus CGMCC 2955 was 5.97 g BC/L (dry weight) when using glycerol as the sole carbon source. Metabolic flux analysis for the central carbon metabolism revealed that about 47.96 % of glycerol was transformed into BC, while only 19.05 % of glucose and 24.78 % of fructose were transformed into BC. Instead, when glucose was used as the sole carbon source, 40.03 % of glucose was turned into the by-product gluconic acid. Compared with BC from glucose and fructose, BC from the glycerol medium showed the highest tensile strength at 83.5 MPa, with thinner fibers and lower porosity. As a main byproduct of biodiesel production, glycerol holds great potential to produce BC with superior mechanical and microstructural characteristics.
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Celulose/biossíntese , Gluconacetobacter xylinus/metabolismo , Celulose/química , Meios de Cultura/metabolismo , Frutose/metabolismo , Gluconacetobacter xylinus/química , Glucose/metabolismo , Glicerol/metabolismo , Análise do Fluxo MetabólicoRESUMO
High water cut of produced fluid is one of the most common problems in reservoir development. At present, injecting plugging agents and other profile control and water plugging technologies are the most widely used solutions. With the development of deep oil and gas resources, high-temperature and high-salinity (HTHS) reservoirs are becoming increasingly common. Conventional polymers are prone to hydrolysis and thermal degradation under HTHS conditions, making polymer flooding or polymer-based gels less effective. Phenol-aldehyde crosslinking agent gels can be applied to different reservoirs with a wide range of salinity, but there exist the disadvantage of high cost of gelants. The cost of water-soluble phenolic resin gels is low. Based on the research of former scientists, copolymers consisting of acrylamide (AM) and 2-Acrylamido-2-Methylpropanesulfonic acid (AMPS) and modified water-soluble phenolic resin were used to prepare gels in the paper. The experimental results show that the gelant with 1.0 wt% AM-AMPS copolymer (AMPS content is 47%), 1.0 wt% modified water-soluble phenolic resin and 0.4 wt% thiourea has gelation time of 7.5 h, storage modulus of 18 Pa and no syneresis after aging for 90 days at 105 °C in simulated Tahe water of 22 × 104 mg/L salinity. By comprehensively comparing the effectiveness of the gels prepared by a kind of phenolic aldehyde composite crosslinking agent and modified water-soluble phenolic resin, it is found that the gel constructed by the modified water-soluble phenolic resin not only reduces costs, but also has shorter gelation time and higher gel strength. The oil displacement experiment with a visual glass plate model proves that the forming gel has good plugging ability and thus improves the sweep efficiency. The research expands the application range of water-soluble phenolic resin gels, which has an important implication for profile control and water plugging in the HTHS reservoirs.
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As a temperature-resistant and salt-resistant polymer, acrylamide and 2-acrylamide-2-methylpropane sulfonic acid (abbreviated as AM-AMPS) copolymer is currently widely used in drilling, water control and oil production stabilization, enhanced oil recovery and other fields, but its stability under high temperature has been less studied. The degradation process of the AM-AMPS copolymer solution was studied by measuring viscosity, the degree of hydrolysis, and weight-average molecular weight at different temperatures and aging time. During the high-temperature aging process, the viscosity of the AM-AMPS copolymer saline solution first increases and then decreases. The combined action of the hydrolysis reaction and the oxidative thermal degradation leads to the change of the viscosity of the AM-AMPS copolymer saline solution. The hydrolysis reaction of the AM-AMPS copolymer mainly affects the structural viscosity of its saline solution through intramolecular and intermolecular electrostatic interactions, while the oxidative thermal degradation mainly reduces its molecular weight by breaking the main chain of the copolymer molecules, reducing the viscosity of the AM-AMPS copolymer saline solution. The content of AM and AMPS groups in the AM-AMPS copolymer solution at various temperatures and aging time was analyzed using liquid nuclear magnetic resonance carbon spectroscopy, demonstrating that the hydrolysis reaction rate constant of AM groups was significantly higher than that of AMPS groups. The contribution values of hydrolysis reaction and oxidative thermal degradation of the AM-AMPS copolymer at different aging time to viscosity were quantitatively calculated at temperatures ranging from 104.5 °C to 140 °C. It was determined that the higher the heat treatment temperature, the smaller the contribution of hydrolysis reaction to viscosity, while the bigger the contribution of oxidative thermal degradation to the viscosity of the AM-AMPS copolymer solution.
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Polymer gel plugging is an effective technique for gas mobility control in flue gas flooding reservoirs. However, the performance of polymer gels is extremely susceptible to the injected flue gas. A reinforced chromium acetate/partially hydrolyzed polyacrylamide (HPAM) gel, using thiourea as the oxygen scavenger and nano-SiO2 as the stabilizer, was formulated. The related properties were evaluated systematically, including gelation time, gel strength, and long-term stability. The results indicated that the degradation of polymers was effectively suppressed by oxygen scavengers and nano-SiO2. The gel strength would be increased by 40% and the gel kept desirable stability after aging for 180 days at elevated flue gas pressures. Dynamic light scattering (DLS) analysis and Cryo-scanning electron microscopy (Cryo-SEM) revealed that nano-SiO2 was adsorbed on polymer chains by hydrogen bonding, which improved the homogeneity of gel structure and thus enhanced the gel strength. Besides, the compression resistance of gels was studied by creep and creep recovery tests. The failure stress of gel with the addition of thiourea and nanoparticles could reach up to 35 Pa. The gel retained a robust structure despite extensive deformation. Moreover, the flow experiment indicated that the plugging rate of reinforced gel still maintained up to 93% after flue gas flooding. It is concluded that the reinforced gel is applicable for flue gas flooding reservoirs.
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Senecavirus A (SVA) is a type of nonenveloped single-stranded, positive-sense RNA virus. The VP2 protein is a structural protein that plays an important role in inducing early and late immune responses of the host. However, its antigenic epitopes have not been fully elucidated. Therefore, defining the B epitopes of the VP2 protein is of great importance to revealing its antigenic characterization. In this study, we analyzed B-cell immunodominant epitopes (IDEs) of the VP2 protein from the SVA strain CH/FJ/2017 using the Pepscan approach and a bioinformatics-based computational prediction method. The following four novel IDEs of VP2 were identified: IDE1, 41TKSDPPSSSTDQPTTT56; IDE2, 145PDGKAKSLQELNEEQW160; IDE3, 161VEMSDDYRTGKNMPF175; and IDE4, 267PYFNGLRNRFTTGT280. Most of the IDEs were highly conserved among the different strains. To our knowledge, the VP2 protein is a major protective antigen of SVA that can induce neutralizing antibodies in animals. Here, we analyzed the immunogenicity and neutralization activity of four IDEs of VP2. Consequently, all four IDEs showed good immunogenicity that could elicit specific antibodies in guinea pigs. A neutralization test in vitro showed that the peptide-specific guinea pig antisera of IDE2 could neutralize SVA strain CH/FJ/2017, and IDE2 was identified as a novel potential neutralizing linear epitope. This is the first time VP2 IDEs have been identified by using the Pepscan method and a bioinformatics-based computational prediction method. These results will help elucidate the antigenic epitopes of VP2 and clarify the basis for immune responses against SVA. IMPORTANCE The clinical symptoms and lesions caused by SVA are indistinguishable from those of other vesicular diseases in pigs. SVA has been associated with recent outbreaks of vesicular disease and epidemic transient neonatal losses in several swine-producing countries. Due to the continuing spread of SVA and the lack of commercial vaccines, the development of improved control strategies is urgently needed. The VP2 protein is a crucial antigen on the capsids of SVA particles. Furthermore, the latest research showed that VP2 could be a promising candidate for the development of novel vaccines and diagnostic tools. Hence, a detailed exploration of epitopes in the VP2 protein is necessary. In this study, four novel B-cell IDEs were identified using two different antisera with two different methods. IDE2 was identified as a new neutralizing linear epitope. Our findings will help in the rational design of epitope vaccines and further understanding of the antigenic structure of VP2.
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Proteínas do Capsídeo , Epitopos de Linfócito B , Animais , Cobaias , Proteínas do Capsídeo/genética , Epitopos de Linfócito B/genética , Anticorpos Antivirais , Soros ImunesRESUMO
Polymer gels have been widely used in high water cut oilfields for profile control and water plugging. It is urgent to develop a gel suitable for the Tahe Oilfield (Temperature: 130 °C, salinity: 2.2 × 105 mg/L) in China. A stable gel was prepared by using an acrylamide (AM)/2-acrylamide-2-methyl propanesulfonic acid (AMPS) copolymer crosslinked with urotropin (HMTA), hydroquinone (HQ), thiourea and Nano-SiO2. This paper covers a step-by-step process for designing gels based on experience with preparing gels. A wide range of combinations between polymers and crosslinking agents with and without stabilizers were investigated, and the results indicated that there is an optimal value of AMPS content of AM/AMPS copolymers in the preparation of gels. Increasing the mass fraction of copolymer and using stabilizer enhanced the performance of gel, but an excessive amount of crosslinking agent was not conducive to the stability of gel. The work optimized the formula of plugging agent suitable for the high temperature and high salt (HTHS) condition in the Tahe Oilfield. The gelling solution had a long gelation time of 20 h. The gel had high strength (Sydansk's gel-strength code of "G") with storage modulus of 12.9 Pa and could be stable for half a year at 130 °C and 2.2 × 105 mg/L of salinity. The plate model that could be heated and pressurized was used to simulate the oil flooding and profile modification under the condition of the Tahe Oilfield for the first time. The experiment results showed that the oil recovery could be increased by 13.22% by subsequent water flooding under heterogeneous formation condition. Therefore, it was fully confirmed that the plugging performance of AM/AMPS phenolic gel prepared in the work was excellent. The information provided in the study could be used as a reference for the design and evaluation of polymer gels in other HTHS reservoirs.
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HYPOTHESIS: Poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) could be adsorbed on the silica surface via the hydrogen bonding between PEO and silanol (SiOH) groups. This interaction would be inhibited once SiOH is dissociated to SiO- at an increased pH value. Besides, the adsorption should be affected by temperature considering the nature of hydrogen bond. Hence, we speculate that silica nanoparticles modified in situ by adsorbed PEO-PPO-PEO possess a pH- and thermo-sensitive surface activity, making them a stimuli-responsive Pickering emulsifier. EXPERIMENTS: Paraffin oil-in-water emulsions stabilized by silica nanoparticles and PEO-PPO-PEO were prepared. Stabilities, droplet morphologies and stimuli-responses were systematically studied using bottle test, optical microscopy and cryo-scanning electron microscopy (cryo-SEM). To clarify the emulsification mechanism, interfacial viscoelastic moduli and desorption energies were determined using the data obtained from drop shape analysis. FINDINGS: Silica nanoparticles are hydrophobized and flocculated by adsorbed PEO-PPO-PEO at a relatively low pH and room temperature. Upon the pH or the temperature increased, particles regain their hydrophilicity and dispersity due to the desorption of surfactants. Emulsions stabilized by these surfactant-modified particles are pH- and thermo-responsive and can be repetitively switched between stabilization and destabilization. The switch temperature is controlled by the PEO length. The emulsification mechanism is verified in view of interfacial viscoelasticity and desorption energy. These findings demonstrate a novel and simple strategy of preparing pH- and thermo-responsive Pickering emulsions desirable to many industrial applications.
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Nanopartículas , Dióxido de Silício , Emulsões/química , Concentração de Íons de Hidrogênio , Nanopartículas/química , Polímeros , Dióxido de Silício/química , Tensoativos/químicaRESUMO
Polymer gel plugging is an effective method for gas mobility control in flue gas flooding reservoirs. However, the effect and mechanism of flue gas on the performance of polymer gels have rarely been reported. In this study, a polymer gel was prepared by cross-linking hydrolyzed polyacrylamide (HPAM) and resorcinol/ hexamethylenetetramine (HMTA) to illuminate the influencing mechanism of flue gas composition on gel. The gel rheological testing results showed that flue gas promoted gelation performance, whereas it seriously threatened gel long-term stability, especially at high pressure conditions. The influence of CO2 on the polymer gel had the characteristic of multiplicity. The hydrodynamic radius (Rh) and the initial viscosity of HPAM solution decreased in the presence of CO2. Nonetheless, the dissolved CO2 expedited the decomposition rate of HMTA into formaldehyde, which promoted the cross-linking process of the HPAM, leading to a shorter gelation time. Oxidation-reduction potential (ORP) tests and Fourier transform infrared spectroscopy (FTIR) analysis indicated that O2 played a leading role in the oxidative degradation of HPAM compared to CO2 and threatened the gel long-term stability at elevated gas pressures. To address the adverse effects caused by flue gas, it is highly desirable to develop polymer gels by adding oxygen scavengers or strengthening additives.
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Ultralow oil-water interfacial tension (IFT) has provided an important basis for screening optimum surfactant formulation for improving the oil washing efficiency. Thus, it is of great significance to further investigate the selection method for surfactant systems with ultralow IFT. In this study, a selection of surfactant systems with ultralow IFT was simplified by a method of comparing the equivalent alkane carbon number (EACN) of crude oil with the minimum alkane carbon number (n min) of surfactant mixtures. The results show that the ultralow IFT can be achieved when the n min of optimum surfactant formulation is equal to the EACN of crude oil. Meanwhile, the oil washing efficiency experiments show that the oil washing efficiency increases with the decrease of IFT, and the optimum surfactant formulation with ultralow IFT has the highest oil washing efficiency. This study provides a more efficient way for selecting optimum surfactant formulation systems with ultralow IFT for improving the oil washing efficiency.
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Extended surfactants with the oxypropylene (PO) group and ethoxylated anionic surfactants with the oxyethylene (EO) group have a high salt tolerance capability. Most of the researches on extended surfactants and ethoxylated anionic surfactants focused on the microemulsion, solubilization, and interfacial tension (IFT) of concentrated surfactant solutions, whereas a few researches focused on the IFT of dilute surfactant solutions. Moreover, a previous work focused only on surfactants with PO numbers greater than 4 and copolymers of PO and EO. The effects of extended surfactants containing short PO chains and no EO groups have not been examined. We measured the IFT and optimal salinity between n-alkanes and dilute solutions of extended surfactants or ethoxylated sulfonates at 30 °C. The effects of the surfactant structure on the equilibrium interfacial tension (IFTeq) and optimal salinity of the system were studied in detail. As for the effects on IFT, results indicate that the introduction of PO groups leads to their enhanced capability to reduce the IFT prior to cross-salinity and a reduction in the IFT between n-alkanes and surfactant solutions to ultralow values (smaller than 0.01 mN/m) near the optimal salinity. It was also found that extended surfactants with different alkyl chains also entail a cross-salinity; at values lower than the cross-salinity, the IFT reduction capacity of extended surfactants with a long alkyl chain (C16P3SO3) is better than that of extended surfactants with a short alkyl chain (C13P3SO3). As for the effect on the optimal salinity, it was found that the optimal salinity of extended surfactants is lower than that of ethoxylated sulfonates for the same oil phase. It was also found that the optimal salinity of extended surfactants first increased and later decreased with increasing PON. This finding is first proposed based on summarizing some researchers' studies and our experiments.
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The currently accepted approaches to treatment of pulmonary fibrosis are based on the treatment of alveolitis and pulmonary fibrosis, however clinically available anti-inflammatory and antifibrotic agents are not often beneficial. Hu-qi-yin, one of the traditional Chinese herbal formulas, has been used for clinical therapy of pulmonary fibrosis in China. The aim of the present study is to evaluate the preventive effects of Hu-qi-yin on the bleomycin (BLM)-induced pulmonary fibrosis in rats. The degree of fibrosis was evaluated with hydroxyproline contents in serum and lung tissue 28 days post-BLM instillation. The semi-quantitative analyses of lung sections were conducted to evaluate the intensity of alveolitis and fibrosis. Furthermore, the expression of transforming growth factor-beta(1) (TGF-beta(1)) was quantitatively studied at the protein and mRNA levels by immunohistochemistry and in situ hybridization, respectively. Oral treatment with Hu-qi-yin improved the body weight loss of rats in doses of 3.8, 7.6g/kg compared with BLM-treated control group, and significantly inhibited the alveolitis and pulmonary fibrosis in a dose-dependent manner as reflected by decreases of the hydroxyproline contents of serum and lung, and amelioration of alveolitis and pulmonary fibrosis fraction 28 days after BLM administration. The one of the possible mechanisms of the protective effect of Hu-qi-yin was via reduction of the overexpression of TGF-beta(1) protein and mRNA. These results suggested a great potential that Hu-qi-yin might be effective in the treatment of pulmonary fibrosis.
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Medicina Herbária , Fibrose Pulmonar/tratamento farmacológico , Administração Oral , Animais , Bleomicina , Peso Corporal/efeitos dos fármacos , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Técnica Indireta de Fluorescência para Anticorpo , Hidroxiprolina/sangue , Hidroxiprolina/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Pulmão/patologia , Masculino , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/patologia , RNA Mensageiro/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Organismos Livres de Patógenos Específicos , Fatores de Tempo , Fator de Crescimento Transformador beta1/análise , Fator de Crescimento Transformador beta1/metabolismoRESUMO
Anxiety and depression are two common psychological disorders with high morbidity worldwide. Understanding of their prevalence of patients with chronic respiratory diseases is becoming more and more important for clinicians. The current study aims at investigation of the prevalence and potential risk factors of anxiety and depression among patients with chronic respiratory diseases. The psychological status, anxiety and depression, and the relevant risk factors of 1713 patients with chronic respiratory diseases from 8 general hospitals in Jiangsu Province of China were evaluated. The results showed that the patients with chronic respiratory diseases experiencing depression and anxiety accounted for 46.00% and 25.34%, respectively. Multivariate logistic regression analysis revealed that lower body mass index (BMI), sleep disorders, limitation of physical activity, and negative life events were significantly associated with an increased risk of both depressive and anxiety symptoms. Poor marital status including divorce, separation and widowerhood was markedly correlated with an increased risk of depression, and chronic pain was with anxiety symptoms, respectively. Collectively, the data demonstrated that depression and anxiety were highly prevalent among the patients with chronic respiratory diseases. It is greatly significant to take specific psychological measures to lower the incidence of depression and anxiety in these patients.
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Ansiedade/epidemiologia , Depressão/epidemiologia , Transtornos Respiratórios/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Ansiedade/psicologia , China/epidemiologia , Comorbidade , Estudos Transversais , Depressão/psicologia , Feminino , Hospitais Gerais , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Adulto JovemRESUMO
Although PEGylation is a potential approach to prolong the half-lives and reduce the dosing frequency of therapeutic proteins, conjugation behaviors of polymer have pivotal effects on the remaining bioactivities of the derivatives. In this study, the PEGylation strategy of recombinant human interleukin-1 receptor antagonist was investigated. The random conjugation of polyethylene glycol to amino groups on the protein resulted in a severe loss of activity and only retained 9.8% of the activity. In contrast, the PEGylation at the thiol groups had moderate effects on the bioactivity of protein and 40% of activity was conserved. The results suggested that the thiol-target PEGylation was more beneficial for IL-1ra.