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Rheumatoid arthritis (RA) progression involves multiple cell types, and sequential drug action on target cells is necessary for RA treatment. Nanocarriers are widely used for RA treatment; however, the targeted delivery and on-demand release of multiple drugs remains challenging. Therefore, in this study, a dual-sensitive polymer is developed using chondroitin sulfate (CS) for the co-delivery of the cartilage repair agent, glucosamine (GlcN), and anti-inflammatory drug, tofacitinib (Tof). In the joint cavity, acidic pH facilitates the cleavage of GlcN from CS polymer to repair the cartilage damage. Subsequently, macrophage uptake via CS-CD44 binding and intracellular reactive oxygen species (ROS) mediate conversion of (methylsulfanyl)propylamine to a hydrophilic segment jointly triggered rapid Tof/GlcN release via micelle disassembly. The combined effects of Tof, GlcN, and ROS depletion promote the M1-to-M2 polarization shift to attenuate inflammation. The synergistic effects of these agents against RA are confirmed in vitro and in vivo. Overall, the dual pH/ROS-sensitive CS nanoplatform simultaneously delivers GlcN and Tof, providing a multifunctional approach for RA treatment with synergistic drug effects.
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Artrite Reumatoide , Glucosamina , Piperidinas , Pirimidinas , Espécies Reativas de Oxigênio , Espécies Reativas de Oxigênio/metabolismo , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/metabolismo , Piperidinas/química , Piperidinas/farmacologia , Concentração de Íons de Hidrogênio , Glucosamina/química , Animais , Pirimidinas/química , Pirimidinas/farmacologia , Camundongos , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Sinergismo Farmacológico , Nanopartículas/química , Células RAW 264.7 , HumanosRESUMO
Spring viremia of carp virus (SVCV) is a highly pathogenic Vesiculovirus in the common carp. The phosphoprotein (P protein) of SVCV is a multifunctional protein that acts as a polymerase cofactor and an antagonist of cellular interferon (IFN) response. Here, we report the 1.5-Å-resolution crystal structure of the P protein central domain (PCD) of SVCV (SVCVPCD). The PCD monomer consists of two ß sheets, an α helix, and another two ß sheets. Two PCD monomers pack together through their hydrophobic surfaces to form a dimer. The mutations of residues on the hydrophobic surfaces of PCD disrupt the dimer formation to different degrees and affect the expression of host IFN consistently. Therefore, the oligomeric state formation of the P protein of SVCV is an important mechanism to negatively regulate host IFN response.IMPORTANCE SVCV can cause spring viremia of carp with up to 90% lethality, and it is the homologous virus of the notorious vesicular stomatitis virus (VSV). There are currently no drugs that effectively cure this disease. P proteins of negative-strand RNA viruses (NSVs) play an essential role in many steps during the replication cycle and an additional role in immunosuppression as a cofactor. All P proteins of NSVs are oligomeric, but the studies on the role of this oligomerization mainly focus on the process of virus transcription or replication, and there are few studies on the role of PCD in immunosuppression. Here, we present the crystal structure of SVCVPCD A new mechanism of immune evasion is clarified by exploring the relationship between SVCVPCD and host IFN response from a structural biology point of view. These findings may provide more accurate target sites for drug design against SVCV and provide new insights into the function of NSVPCD.
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Fosfoproteínas/química , Rhabdoviridae/química , Proteínas Virais/química , Animais , Cristalografia por Raios X , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha betaRESUMO
p53, which regulates cell-cycle arrest and apoptosis, is a crucial target for viruses to release cells from cell-cycle checkpoints or to protect cells from apoptosis for their own benefit. Viral evasion mechanisms of aquatic viruses remain mysterious. Here, we report the spring viremia of carp virus (SVCV) degrading and stabilizing p53 in the ubiquitin-proteasome pathway by the N and P proteins, respectively. Early in an SVCV infection, significant induction was observed in the S phase and p53 was decreased in the protein level. Further experiments demonstrated that p53 interacted with SVCV N protein and was degraded by suppressing the K63-linked ubiquitination. However, the increase of p53 was observed late in the infection and experiments suggested that p53 was bound to SVCV P protein and stabilized by enhancing the K63-linked ubiquitination. Finally, lysine residue 358 was the key site for p53 K63-linked ubiquitination by the N and P proteins. Thus, our findings suggest that fish p53 is modulated by SVCV N and P protein in two distinct mechanisms, which uncovers the strategy for the subversion of p53-mediated host innate immune responses by aquatic viruses.
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Rhabdoviridae/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Animais , Pontos de Checagem do Ciclo Celular/fisiologia , Vírus de DNA , Doenças dos Peixes , Regulação Viral da Expressão Gênica/genética , Células HEK293 , Humanos , Imunidade Inata , Rhabdoviridae/patogenicidade , Ubiquitinação , Viremia , Peixe-Zebra , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismoRESUMO
Viral infection activates the transcription factor IFN regulatory factor 7 (IRF7), which plays a critical role in the induction of IFNs and innate antiviral immune response. How virus-induced IFN signaling is controlled in fish is not fully understood. In this study, we demonstrate that N-myc downstream-regulated gene 1a (NDRG1a) in zebrafish plays a role as a negative regulator for virus-triggered IFN induction. First, the activation of the IFN promoter stimulated by the polyinosinic-polycytidylic acid or spring viremia of carp virus was decreased by the overexpression of NDRG1a. Second, NDRG1a interacted with IRF7 and blocked the IFN transcription activated by IRF7. Furthermore, NDRG1a was phosphorylated by TANK-binding kinase 1 (TBK1) and promoted the K48-linked ubiquitination and degradation of IRF7. Finally, the overexpression of NDRG1a blunted the transcription of several IFN-stimulated genes, resulting in the host cells becoming susceptible to spring viremia of carp virus infection. Our findings suggest that fish NDRG1a negatively regulates the cellular antiviral response by targeting IRF7 for ubiquitination and degradation, providing insights into the novel role of NDRG1a on the innate antiviral immune response in fish.
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Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Fatores Reguladores de Interferon/metabolismo , Interferons/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/veterinária , Rhabdoviridae/fisiologia , Proteínas Supressoras de Tumor/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/imunologia , Animais , Células Cultivadas , Suscetibilidade a Doenças , Imunidade Inata , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteólise , Proteínas Proto-Oncogênicas/metabolismo , Transdução de Sinais , Proteínas Supressoras de Tumor/genética , Ubiquitinação , Proteínas de Peixe-Zebra/genéticaRESUMO
Interferon (IFN) production activated by phosphorylated interferon regulatory factor 7 (IRF7) is a pivotal process during host antiviral infection. For viruses, suppressing the host IFN response is beneficial for viral proliferation; in such cases, evoking host-derived IFN negative regulators would be very useful for viruses. Here, we report that the zebrafish rapunzel 5 (RPZ5) protein which activated by virus degraded phosphorylated IRF7 is activated by TANK-binding kinase 1 (TBK1), leading to a reduction in IFN production. Upon viral infection, zebrafish rpz5 was significantly upregulated, as was ifn, in response to the stimulation. Overexpression of RPZ5 blunted the IFN expression induced by both viral and retinoic acid-inducible gene I (RIG-I) like-receptor (RLR) factors. Subsequently, RPZ5 interacted with RLRs but did not affect the stabilization of the proteins in the normal state. Interestingly, RPZ5 degraded the phosphorylated IRF7 under TBK1 activation through K48-linked ubiquitination. Finally, the overexpression of RPZ5 remarkably reduced the host cell antiviral capacity. These findings suggest that zebrafish RPZ5 is a negative regulator of phosphorylated IRF7 and attenuates IFN expression during viral infection, providing insight into the IFN balance mechanism in fish.IMPORTANCE The phosphorylation of IRF7 is helpful for host IFN production to defend against viral infection; thus, it is a potential target for viruses to mitigate the antiviral response. We report that the fish RPZ5 is an IFN negative regulator induced by fish viruses and degrades the phosphorylated IRF7 activated by TBK1, leading to IFN suppression and promotion of viral proliferation. These findings reveal a novel mechanism for interactions between the host cell and viruses in the lower vertebrate.
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Doenças dos Peixes/virologia , Imunidade Inata/imunologia , Interferons/metabolismo , Infecções por Rhabdoviridae/veterinária , Rhabdoviridae/imunologia , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/virologia , Animais , Antivirais/imunologia , Antivirais/metabolismo , Doenças dos Peixes/imunologia , Doenças dos Peixes/metabolismo , Fatores Reguladores de Interferon/genética , Fatores Reguladores de Interferon/metabolismo , Interferons/imunologia , Fosforilação , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/metabolismo , Infecções por Rhabdoviridae/virologia , Ubiquitinação , Replicação Viral , Peixe-Zebra/fisiologia , Proteínas de Peixe-Zebra/genéticaRESUMO
Fibroblast growth factor receptor (FGFR) 3 is one of the four distinct membrane-spanning tyrosine kinases required for proper skeletal development. In fish, the role of FGFR3 is still unclear. In this article, we reveal that zebrafish FGFR3 is a negative regulator of interferon (IFN) production in the innate immune response by suppressing the activity of TANK-binding kinase 1 (TBK1) in the process of virus infection. qPCR experiments demonstrate that the transcriptional level of cellular FGFR3 was upregulated by infection with spring viremia of carp virus (SVCV), indicating that FGFR3 might be involved in the process of host cell response to viral infection. Then, overexpression of FGFR3 significantly impeded the IFN promoter activity induced by a stimulator. In addition, the capabilities of a retinoic acid-inducible gene I (RIG-I)-like receptor (RLR) system to activate IFN promoter were decreased during the overexpression of FGFR3. Subsequently, FGFR3 decreased the phosphorylation of interferon regulatory factor 3 (IRF3) and mediator of IRF3 activation (MITA) by TBK1. These findings suggest that zebrafish FGFR3 is a negative regulator of IFN by attenuating the kinase activity of TBK1, leading to the suppression of IFN expression.
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Doenças dos Peixes/imunologia , Imunidade Inata/genética , Interferons/genética , Proteínas Serina-Treonina Quinases/genética , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/genética , Proteínas de Peixe-Zebra/genética , Peixe-Zebra/imunologia , Animais , Interferons/metabolismo , Proteínas Serina-Treonina Quinases/imunologia , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/imunologia , Rhabdoviridae/fisiologia , Infecções por Rhabdoviridae/imunologia , Infecções por Rhabdoviridae/veterinária , Transdução de Sinais/imunologia , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/imunologia , Proteínas de Peixe-Zebra/fisiologiaRESUMO
Wireless sensor networks (WSNs) are widely applied in industrial application with the rapid development of Industry 4.0. Combining with centralized cloud platform, the enormous computational power is provided for data analysis, such as strategy control and policy making. However, the data analysis and mining will bring the issue of privacy leakage since sensors will collect varieties of data including sensitive location information of monitored objects. Differential privacy is a novel technique that can prevent compromising single record benefits. Geospatial data can be indexed by a tree structure; however, existing differentially private release methods pay no attention to the concrete analysis about the partition granularity of data domains. Based on the overall analysis of noise error and non-uniformity error, this paper proposes a data domain partitioning model, which is more accurate to choose the grid size. A uniform grid release method is put forward based on this model. In order to further reduce the errors, similar cells are merged, and then noise is added into the merged cells. Results show that our method significantly improves the query accuracy compared with other existing methods.
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The purpose of this study was to investigate the gastrointestinal stability of exenatide to determine the key factor(s) contributing to peptide degradation during the oral delivery process. The effects of pH and various digestive enzymes on the degradation kinetics of exenatide were determined. Moreover, the degradation clearances of peptide were also examined using rat everted intestinal rings and intestinal homogenates from various intestinal locations. Exenatide was comparatively stable within a pH range of 1.2-8. However, obvious degradation was observed in the presence of digestive enzymes. The order of enzymes, in terms of ability to degradate exenatide, was chymotrypsin>aminopeptidase N>carboxypeptidase A>trypsin>pepsin. Chymotrypsin showed the greatest ability to degrade exenatide (half-life t1/2, 5.784×10(-2) h), whereas aminopeptidase N and carboxylpeptidase A gave t1/2 values of 3.53 and 10.16 h, respectively. The degradation of exenatide was found to be peptide concentration- and intestinal site-dependent, with a lower clearance in the upper part of the duodenum and the lower part of the ileum. When using intestinal homogenates as enzyme sources, the order, in terms of peptide degradation ability, was ileum>jejunum>duodenum. However, no significant difference was observed in the remaining peptide concentrations throughout 2 h of incubation, which may be due to the involvement of cytosolic enzymes. These results revealed key factors contributing to peptide degradation, and suggest that the inhibition of chymotrypsin and site-specific delivery of exenatide might be advantageous in overcoming metabolic obstacles during its oral delivery.
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Enzimas/metabolismo , Intestinos/fisiologia , Peptídeos/metabolismo , Peçonhas/metabolismo , Animais , Exenatida , Concentração de Íons de Hidrogênio , Masculino , Peptídeos/química , Ratos , Ratos Wistar , Peçonhas/químicaRESUMO
Solid fuel use is increasingly linked to low birth weight (LBW), but conclusions were inconsistent. We aimed to summarize the association between solid fuel use and LBW. Twenty-one studies that met the inclusion criteria were identified through PubMed, Qvid Medline, and Web of Science databases. The final search occurred on March 20, 2024. Summary relative effect and 95â¯% confidence intervals were estimated with a random-effects model. Subgroup analyses and sensitivity analyses were performed to investigate possible sources of heterogeneity and to test the stability of the results. Nineteen studies evaluated the association between solid fuel use in pregnant woman and LBW (1.188 for solid fuels: 1.055 to 1.322). No significant heterogeneity was identified among the included studies (p=0.010, Tau2=0.02, I2=48.1â¯%). Subgroup analysis found positive correlations for Asia, data years prior to 2014, and rural studies (1.245 for Asia: 1.077 to 1.412; Tau2=0.03, I2=56.0â¯%; 1.243 for data years prior to 2014: 1.062 to 1.424; Tau2=0.04, I2=60.98â¯%; 1.514 for rural: 1.258 to 1.771; Tau2=0.00, I2=0.0â¯%). Our meta-analysis showed that solid fuel use in pregnant women had an impact on LBW. Measures and policies are also needed to promote energy conversion and to limit and reduce the use of solid fuels.
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Significant advancements have been made in catalytic asymmetric α-C-H bond functionalization of ethers via carbenoid insertion over the past decade. Effective asymmetric catalytic systems, featuring a range of chiral metal catalysts, have been established for the enantioselective synthesis of diverse ether substrates. This has led to the generation of various enantioenriched, highly functionalized oxygen-containing structural motifs, facilitating their application in the asymmetric synthesis of bioactive natural products.
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Internet of Things realizes the ubiquitous connection of all things, generating countless time-tagged data called time series. However, real-world time series are often plagued with missing values on account of noise or malfunctioning sensors. Existing methods for modeling such incomplete time series typically involve preprocessing steps, such as deletion or missing data imputation using statistical learning or machine learning methods. Unfortunately, these methods unavoidable destroy time information and bring error accumulation to the subsequent model. To this end, this paper introduces a novel continuous neural network architecture, named Time-aware Neural-Ordinary Differential Equations (TN-ODE), for incomplete time data modeling. The proposed method not only supports imputation missing values at arbitrary time points, but also enables multi-step prediction at desired time points. Specifically, TN-ODE employs a time-aware Long Short-Term Memory as an encoder, which effectively learns the posterior distribution from partial observed data. Additionally, the derivative of latent states is parameterized with a fully connected network, thereby enabling continuous-time latent dynamics generation. The proposed TN-ODE model is evaluated on both real-world and synthetic incomplete time-series datasets by conducting data interpolation and extrapolation tasks as well as classification task. Extensive experiments show the TN-ODE model outperforms baseline methods in terms of Mean Square Error for imputation and prediction tasks, as well as accuracy in downstream classification task.
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In this work, polyamide 66/polyphenylene oxide (PA66/PPO) composites, including the flame retardants 98 wt% aluminum diethylphosphinate + 2 wt% polydimethylsiloxane (P@Si), Al(OH)3-coated red phosphorus (RP*), and glass fiber (GF), were systematically studied, respectively. The limiting oxygen index (LOI), UL-94 vertical burning level, and thermal and mechanical properties of the PA66/PPO composites were characterized. The results showed that the P@Si and RP flame retardants both improved the LOI value and UL-94 vertical burning level of the PA66/PPO composites, and PA66/PPO composites passed to the UL-94 V-0 level when the contents of P@Si and RP* flame retardants were 16 wt% and 8 wt%. On the other hand, the mechanical properties of the PA66/PPO composites were reduced from a ductile to a brittle fracture mode. The addition of GF effectively made up for these defects and improved the mechanical properties of the PA66/PPO composites containing the P@Si and RP*, but it did not change the fracture mode. P@Si and RP* flame retardants improved the thermal decomposition of PA66/PPO/GF composites and reduced the maximum mass loss rates, showing that the PA66/PPO/GF composites containing the P@Si and RP* flame retardants could be used in higher-temperature fields.
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The sugars will eventually be exported transporters (SWEETs) gene family is a new type of sugar transporters, which plays an important role in plant growth and development, physiological metabolism, and abiotic stress. In this study, we used quantitative real-time PCR to analyze the expression of ZmSWEET15a gene in different organs of maize and under different abiotic stresses. The results showed that ZmSWEET15a was expressed in roots, stems, leaves, and grains, with the highest expression level in leaves, which was highly correlated with leaf development. Under the treatment of polyethylene glycol (PEG), NaCl, H2O2, and abscisic acid stress, the expression of ZmSWEET15a was upregulated, while under the treatment of cold stress, the expression of ZmSWEET15a was inhibited. In sugar-specific experiments, we found that sucrose was the most effective carbon source for maize seed germination. The expression analysis of ZmSWEET15a in different carbon sources suggested that the expression of ZmSWEET15a was more likely to be induced by sucrose. Overexpression of ZmSWEET15a in maize plants could reduce the sucrose content in leaves and increase the sucrose content in grains. The heterologous expression of ZmSWEET15a in the yeast mutant strain SUSY7/ura indicated that ZmSWEET15a is a sucrose transporter and pH independent. This study provides new insight into sugar transport and carbohydrate partitioning in maize and other crops, and provide more genetic information for improving crop quality at the molecular level.
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Regulação da Expressão Gênica de Plantas , Zea mays , Carbono/metabolismo , Peróxido de Hidrogênio , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico , Sacarose/metabolismo , Açúcares/metabolismo , Zea mays/genética , Zea mays/metabolismoRESUMO
As environmental concerns began to emerge, companies started to target toward the growing 'green market' to launch their green products. Companies' green advertising played an important role in facilitating corporate green marketing and fuelling the desire for environmental-friendly commodities. Applying a Critical Discursive Perspective, this study focuses on corporate environmental advertising in order to illuminate their discursive strategies and the process through which corporate green advertising generates and symbolically structures the necessity of green consumption. The comparison of constructive characteristics and constructed meanings of green advertisings identified embody distinction in Western and Chinese cognitive style and process.
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Insulin-degrading enzyme (IDE) is a protease that has been demonstrated to play a key role in degrading both Abeta and insulin and deficient in IDE function is associated with Alzheimer's disease (AD) and type 2 diabetes mellitus (DM2) pathology. However, little is known about the cellular and molecular regulation of IDE expression. Here we show IDE levels are markedly decreased in DM2 patients and positively correlated with the peroxisome proliferator-activated receptor gamma (PPARgamma) levels. Further studies show that PPARgamma plays an important role in regulating IDE expression in rat primary neurons through binding to a functional peroxisome proliferator-response element (PPRE) in IDE promoter and promoting IDE gene transcription. Finally, we demonstrate that PPARgamma participates in the insulin-induced IDE expression in neurons. These results suggest that PPARgamma transcriptionally induces IDE expression which provides a novel mechanism for the use of PPARgamma agonists in both DM2 and AD therapies.
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Insulisina/genética , Neurônios/enzimologia , PPAR gama/metabolismo , Adulto , Idoso , Doença de Alzheimer/enzimologia , Animais , Sequência de Bases , Diabetes Mellitus Tipo 2/enzimologia , Feminino , Hipocampo/efeitos dos fármacos , Hipocampo/enzimologia , Humanos , Insulina/farmacologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Neurônios/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Transcrição GênicaRESUMO
Drawing on psychological contract theory, this research contributes to the unethical behavior literature by exploring employees' turnover intentions as a mediator of the relationship between abusive supervision and employees' unethical behavior and the moderating role of the caring climate in the relationship between turnover intentions and unethical behavior. The results from a sample of 679 reveal that turnover intentions mediate relationship between abusive supervision and subordinates' unethical behavior, and caring climate moderates the positive relationship between turnover intentions and subordinates' unethical behavior. We also find that the indirect effect is moderated by the caring climate. The theoretical and practical implications are discussed.
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Pessoal Administrativo/psicologia , Bullying/ética , Bullying/psicologia , Empatia/ética , Instalações Industriais e de Manufatura , Reorganização de Recursos Humanos/estatística & dados numéricos , Local de Trabalho/psicologia , Adulto , China , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Zein-based carriers are a promising delivery system for biomedical applications. However, few studies involve systematic investigation on their in vivo biocompatibility and immunogenicity. PURPOSE: The objective of this study was to identify the immunogenicity, type of immune response, biocompatibility and systemic recall immune response of zein nanoparticles administrated via different routes in mice. ANIMALS AND METHODS: Female Balb/c mice were selected as the animal model in this paper. The effect of particle size, dose and inoculation routes on immunogenicity were systematically explored. The mice were challenged at week 50 via intramuscular and subcutaneous routes to investigate the systemic recall immune responses of zein nanoparticles. Hematoxylin and eosin staining was performed to investigate the biocompatibility of zein nanoparticles at injection sites. RESULTS: The administration of zein particles by parenteral routes led to a long-term systemic immune response. Particle size did not affect zein-specific IgG antibody titers. IgG antibody titers and inflammatory cell infiltration at the injection sites resulting from intramuscular zein particle injection were significantly higher than those from subcutaneous injection of the same dose. For intramuscular inoculation, dose-dependent IgG antibody titers were observed after the third inoculation, while no significant difference was found via the subcutaneous route. For both routes, IgG titer showed a time-dependent decrease at all dose levels from week 5 onward, and finally plateaued at week 28. The IgG subtype assay showed a predominant Th2-type immune response for both administration routes. Challenge with zein nanoparticles at week 50 led to a significant increase in specific IgG titer at all dose levels, indicating systemic recall immune responses. Interestingly, IgG antibody levels in the subcutaneous groups showed a delayed decrease compared to those of the intramuscular injection groups at all dose levels. CONCLUSION: This study indicated that immunogenicity may be one of the key challenges of using zein nanoparticles as carriers via parenteral administration. Further investigation is needed to illustrate zein immunogenicity in other forms, and the possible effect of systemic recall immune response on in vivo pharmacokinetic characteristics.
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Portadores de Fármacos/administração & dosagem , Imunoglobulina G/imunologia , Nanopartículas/administração & dosagem , Zeína/química , Animais , Relação Dose-Resposta Imunológica , Feminino , Imunoglobulina G/sangue , Injeções Intramusculares , Injeções Subcutâneas , Camundongos Endogâmicos BALB C , Nanopartículas/química , Tamanho da Partícula , Células Th2/efeitos dos fármacos , Células Th2/imunologia , VacinaçãoRESUMO
Expression of major histocompatibility complex class II (MHC II) molecules, which determines both the immune repertoire during development and subsequent triggering of immune responses, is always under the control of a unique (MHC class II) transactivator, CIITA. The IFN-γ-inducible MHC II expression has been extensively and thoroughly studied in humans, but not in bony fish. In this study, the characterization of CIITA was identified and its functional domains were analyzed in grass carp. The absence of GAS and E-box in the promoter region of grass carp CIITA, might imply that the cooperative interaction between STAT1 and USF1 to active the CIITA expression, found in mammals, is not present in bony fish. After the transfection of IFN-γ or IFN-γ rel, only IFN-γ could induce MHC II expression mediated by CIITA. Moreover, interferon regulatory factor (IRF) 2, which cooperates with IRF1 to active the CIITA promoter IV expression in mammals, played an antagonistic role to IRF1 in the activation of grass carp CIITA. These data suggested that grass carp, compared with mammals, has both conservative and unique mechanisms in the regulation of MHC II expression.
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Carpas/genética , Carpas/metabolismo , Regulação da Expressão Gênica , Antígenos de Histocompatibilidade Classe II/genética , Fator Regulador 1 de Interferon/metabolismo , Fator Regulador 2 de Interferon/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Carpas/imunologia , Linhagem Celular , DNA Complementar/química , DNA Complementar/genética , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos , Imunomodulação , Interferon gama/metabolismo , Proteínas Nucleares/metabolismo , Fases de Leitura Aberta , Regiões Promotoras Genéticas , Ligação Proteica , Transativadores/metabolismoRESUMO
Zein-based nano/microparticles have been demonstrated to be promising carrier systems for both the food industry and biomedical applications. However, the fabrication of size-controlled zein particles has been a challenging issue. In this study, a modified anti-solvent precipitation method was developed, and the effects of various factors, such as mixing method, solvent/anti-solvent ratio, temperature, zein concentrations and the presence of sodium caseinate (SC) on properties of zein particles were investigated. Evidence is presented that, among the previously mentioned factors, the mixing method, especially mixing rate, could be used as an effective parameter to control the size of zein particles without changing other parameters. Moreover, through fine-tuning the mixing rate together with zein concentration, particles with sizes ranging from nanometers to micrometers and low polydispersity index values could be easily obtained. Based on the size-controlled fabrication method, SC-coated zein nanoparticles could also be obtained in a size-controlled manner by incubation of the coating material with the already-formed zein particles. The resultant nanoparticles showed better performance in both drug loading and controlled release, compared with zein/SC hybrid nanoparticles fabricated by adding aqueous ethanol solution to SC solution. The possible mechanisms of the nanoprecipitation process and self-assembly formation of these nanoparticles are discussed.