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To examine the association between cytomegalovirus (CMV) seropositivity and all-cause mortality in a nationwide cohort of US adults. We obtained data from the National Health and Nutrition Examination Survey III (1988-1994), including 16,547 participants aged 18-90 years old with CMV serology assessments. Mortality status was ascertained until December 2019 using the National Death Index linkage data. The Cox proportional hazard model was applied to estimate the association between CMV seropositivity and mortality. During a median follow-up of 26.3 years, 6,930 deaths were recorded. CMV seropositivity was associated with a higher hazard of all-cause mortality after adjusting for attained age, sex, and ethnicity (HR: 1.22, 95% CI: 1.10, 1.36, p < 0.001). The magnitude of the association attenuated slightly after adjusting further for body mass index, family income, smoking status, diabetes, and self-reported cancer history (HR = 1.11, 95% CI: 1.00, 1.23, p = 0.04). While the association was observed for both men and women, it was only statistically significant among non-Hispanic white people (HR: 1.16, 95% CI: 1.06, 1.26, p = 0.001) but not among other ethnic populations. CMV seropositivity might be an independent risk factor for all-cause mortality among US adults. If the findings are validated in an independent population, further research is needed to unveil the biological mechanisms driving the increased mortality with CMV seropositivity.
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Infecciones por Citomegalovirus , Citomegalovirus , Adulto , Masculino , Humanos , Femenino , Adolescente , Adulto Joven , Persona de Mediana Edad , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Infecciones por Citomegalovirus/complicaciones , Infecciones por Citomegalovirus/epidemiología , Encuestas Nutricionales , Estudios ProspectivosRESUMEN
BACKGROUND: Previous studies have demonstrated associations between fatty acids and neurological disorders. However, no studies have examined the relationship between serum fatty acid levels and serum neurofilament light chain (NfL), a biomarker of neurological disorders. OBJECTIVES: This study aimed to comprehensively investigate the intricate relationship between 30 serum fatty acids and serum NfL levels in a nationally representative sample of United States adults, using data from the 2013-2014 National Health and Nutrition Examination Survey. METHODS: Using a cross-sectional analysis, multivariable linear regression models were used to explore the associations between 30 serum fatty acids and serum NfL levels. This analysis involved adjustment for potential confounding variables, including age, sex, race, body mass index (BMI), smoking status, hyperlipidemia, and diabetes, to clarify the association between serum fatty acids and serum NfL levels. RESULTS: The analysis revealed that certain fatty acids exhibited distinct associations with serum NfL levels. Notably, docosanoic acid (22:0) and tricosanoic acid (C23:0) were found to be inversely associated with serum NfL levels (ß = -0.280, 95% confidence interval [CI]: -0.525, -0.035; ß = -0.292, 95% CI: -0.511, -0.072). Conversely, palmitoleic acid (16:1n-7) demonstrated a positive association with serum NfL levels (ß = 0.125, 95% CI: 0.027, 0.222). Notably, these associations remained significant even after adjustment for potential confounders. CONCLUSIONS: Individuals with high relative concentrations of certain SFA exhibited decreased serum NfL, whereas those with high relative concentrations of certain monounsaturated fatty acids showed increased serum NfL. These findings contribute to a deeper understanding of the potential impact of serum fatty acids on NfL levels, shedding light on novel avenues for further investigation and potential interventions in the context of neurological health.
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BACKGROUND: Previous studies have suggested a connection between impaired olfactory function and an increased risk of rapid eye movement sleep behavior disorder (RBD) in individuals diagnosed with Parkinson's disease (PD). However, there is a gap in knowledge regarding the potential impact of olfactory dysfunction on the long-term patterns of sleep disorders among early PD patients. METHODS: Data from the Parkinson's Progression Markers Initiative program included 589 participants with assessments of sleep disorders using the Epworth Sleepiness Scale (ESS) and RBD Screening Questionnaire (RBDSQ). Olfactory dysfunction at baseline was measured using the University of Pennsylvania Smell Identification Test. Trajectories of sleep disorders over a 5-year follow-up were identified using group-based trajectory modeling, and the relationship between olfactory dysfunction and sleep disorder trajectories was examined through binomial logistic regression. RESULTS: Two distinct trajectories of sleep disorders over the 5-year follow-up period were identified, characterized by maintaining a low or high ESS score and a low or high RBDSQ score. An inversion association was observed between olfactory function measures and trajectories of excessive daytime sleepiness (odds ratio [OR] = 0.97, 95% confidence interval [CI] 0.95, 1.00, p = 0.038), after controlling for potential covariates. Similarly, olfactory function showed a significant association with lower trajectories of probable RBD (OR = 0.96, 95% CI 0.94, 0.98, p = 0.001) among early PD individuals. Consistent findings were replicated across alternative analytical models. CONCLUSIONS: Our findings indicated that olfactory dysfunction was associated with unfavorable long-term trajectories of sleep disorders among early PD.
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OBJECTIVE: To examine the associations of excessive daytime sleepiness (EDS) and probable rapid eye movement sleep behavior disorder (pRBD), respectively, with impulsive-compulsive behaviors (ICBs) over a 5-year follow-up in patients with early Parkinson's disease (PD). METHODS: The Parkinson's Progression Markers Initiative is a multicenter cohort study based on an ongoing and open-ended registry. Longitudinal associations of sleep disorders with ICB over 5-year follow-up visits were estimated using generalized linear mixed-effects models among PD participants. RESULTS: A total of 825 PD participants were enrolled at baseline. The study sample had a median baseline age of 63.1 (interquartile range: 55.6-69.3) years and comprised 496 (61.5%) men. Among them, 201 (24.9%) had ICB at baseline. In the generalized mixed-effects models, EDS (odds ratio [OR] = 1.09, 95% confidence interval [CI] 1.05, 1.12) and RBD (OR = 1.07, 95% CI 1.03, 1.12) were substantially associated with higher odds of developing ICB over time in PD patients, after multivariate adjustment including age, gender, family history, GDS score, STAI-Y score, MDS-UPDRS part III score, LEDD, and disease duration. Consistent results were observed when stratifying by age at baseline, gender, and PD family history. CONCLUSIONS: The study findings suggest a longitudinal association between EDS and pRBD with an increased risk of developing ICB in patients with PD. The findings emphasize the significance of evaluating and addressing sleep disorders in PD patients as a potential approach to managing ICB.
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Enfermedad de Parkinson , Humanos , Masculino , Femenino , Enfermedad de Parkinson/epidemiología , Enfermedad de Parkinson/complicaciones , Enfermedad de Parkinson/psicología , Persona de Mediana Edad , Anciano , Estudios Prospectivos , Trastorno de la Conducta del Sueño REM/epidemiología , Trastorno de la Conducta del Sueño REM/complicaciones , Trastornos del Sueño-Vigilia/epidemiología , Conducta Compulsiva/epidemiología , Conducta Impulsiva , Trastornos de Somnolencia Excesiva/epidemiología , Estudios de CohortesRESUMEN
As China faces demographic shifts and socioeconomic changes, the burden of Alzheimer's disease (AD) and associated cognitive impairments is increasing dramatically, with significant implications for public health and the economy. This systematic review and meta-analysis aims to provide a comprehensive assessment of the prevalence and incidence of AD across China. Drawing from an extensive search of international and Chinese databases up to August 27, 2023, including PubMed, Embase, and the Cochrane Library, we synthesized data from 105 studies. Our analysis reveals a combined prevalence of AD of 3.48% within a sample of 626,276 elderly individuals and an incidence rate of 7.90 per 1000 person-years. Subgroup and meta-regression analyses highlight age and gender as pivotal factors influencing these epidemiological patterns. Notably, significant heterogeneity exists due to variations in diagnostic criteria and study quality, impacting the comparability of findings. This meta-analysis underscores the need for continued research into demographic and modifiable risk factors influencing AD, while emphasizing standardized reporting practices to address these limitations and improve the understanding of AD's challenge in China.
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Enfermedad de Alzheimer , Anciano , Femenino , Humanos , Masculino , Enfermedad de Alzheimer/epidemiología , China/epidemiología , Incidencia , Prevalencia , Factores de RiesgoRESUMEN
Electrochemical C-N coupling reaction based on carbon dioxide and nitrate have been emerged as a new "green synthetic strategy" for the synthesis of urea, but the catalytic efficiency is seriously restricted by the inherent scaling relations of adsorption energies of the active sites, the improvement of catalytic activity is frequently accompanied by the decrease in selectivity. Herein, a doping engineering strategy was proposed to break the scaling relationship of intermediate binding and minimize the kinetic barrier of C-N coupling. A thus designed SrCo0.39Ru0.61O3-δ catalyst achieves a urea yield rate of 1522â µg h-1 mgcat. -1 and faradic efficiency of 34.1 % at -0.7â V versus reversible hydrogen electrode. A series of characterizations revealed that Co doping not only induces lattice distortion but also creates rich oxygen vacancies (OV) in the SrRuO3. The oxygen vacancies weaken the adsorption of *CO and *NH2 intermediates on the Co and Ru sites respectively, and the strain effects over the Co-Ru dual sites promoting the occurrence of C-N coupling of the two monomers instead of selective hydrogenating to form by-products. This work presents an insight into molecular coupling reactions towards urea synthesis via the doping engineering on SrRuO3.
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Non-noble metal oxides have emerged as potential candidate electrocatalysts for acidic oxygen evolution reactions (OERs) due to their earth abundance; however, improving their catalytic activity and stability simultaneously in strong acidic electrolytes is still a major challenge. In this work, we report Co3O4@carbon core-shell nanoparticles on 2D graphite sheets (Co3O4@C-GS) as mixed-dimensional hybrid electrocatalysts for acidic OER. The obtained Co3O4@C-GS catalyst exhibits a low overpotential of 350 mV and maintains stability for 20 h at a current density of 10 mA cm-2 in H2SO4 (pH = 1) electrolyte. X-ray photoelectron and X-ray absorption spectroscopies illustrate that the higher content of Co3+ sites boosts acidic OER. Operando Raman spectroscopy reveals that the catalytic stability of Co3O4@C nanoparticles during the acidic OER is enhanced by the introduction of graphite sheets. This interface engineering of non-noble metal sites with high valence states provides an efficient approach to boost the catalytic activity and enhance the stability of noble-metal-free electrocatalysts for acidic OER.
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We report Ca2-xIrO4 nanocrystals exhibit record stability of 300 h continuous operation and high iridium mass activity (248 A gIr-1 at 1.5 VRHE) that is about 62 times that of benchmark IrO2. Lattice-resolution images and surface-sensitive spectroscopies demonstrate the Ir-rich surface layer (evolved from one-dimensional connected edge-sharing [IrO6] octahedrons) with high relative content of Ir5+ sites, which is responsible for the high activity and long-term stability. Combining operando infrared spectroscopy with X-ray absorption spectroscopy, we report the first direct observation of key intermediates absorbing at 946 cm-1 (Ir6+âO site) and absorbing at 870 cm-1 (Ir6+OO- site) on iridium-based oxides electrocatalysts, and further discover the Ir6+âO and Ir6+OO- intermediates are stable even just from 1.3 VRHE. Density functional theory calculations indicate the catalytic activity of Ca2IrO4 is enhanced remarkably after surface Ca leaching, and suggest IrOO- and IrâO intermediates can be stabilized on positive charged active sites of Ir-rich surface layer.
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Iridium-based perovskites show promising catalytic activity for oxygen evolution reaction (OER) in acid media, but the iridium mass activity remains low and the active-layer structures have not been identified. Here, we report highly active 1 nm IrOx particles anchored on 9R-BaIrO3 (IrOx/9R-BaIrO3) that are directly synthesized by solution calcination followed by strong acid treatment for the first time. The developed IrOx/9R-BaIrO3 catalyst delivers a high iridium mass activity (168 A gIr-1), about 16 times higher than that of the benchmark acidic OER electrocatalyst IrO2 (10 A gIr-1), and only requires a low overpotential of 230 mV to reach a catalytic current density of 10 mA cm-2geo. Careful scanning transmission electron microscopy, synchrotron radiation-based X-ray absorption spectroscopy, and X-ray photoelectron spectroscopy analyses reveal that, during the electrocatalytic process, the initial 1 nm IrOx nanoparticles/9R-BaIrO3 evolve into amorphous Ir4+OxHy/IrO6 octahedrons and then to amorphous Ir5+Ox/IrO6 octahedrons on the surface. Such high relative content of amorphous Ir5+Ox species derived from trimers of face-sharing IrO6 octahedrons in 9R-BaIrO3 and the enhanced metallic conductivity of the Ir5+Ox/9R-BaIrO3 catalyst are responsible for the excellent acidic OER activity. Our results provide new insights into the surface active-layer structure evolution in perovskite electrocatalysts and demonstrate new approaches for engineering highly active acidic OER nanocatalysts.
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As a new, to the best of our knowledge, alternative to the saturated vapor-cell-based Faraday anomalous dispersion optical filter (FADOF), the FADOF based on a hollow cathode lamp (HCL-FADOF) not only enables the FADOF to work normally at room temperature without heating, but also has some new features due to the inherent characteristics of the HCL. In this Letter, we implement an HCL-FADOF operating on the rubidium D2 line and experimentally investigate the effect of ambient temperature on its performance and cold-start characteristics. Results show that the HCL-FADOF can provide excellent stability within a large temperature range, even at temperatures below 0°C. A comparison of the start performance between the HCL-FADOF and FADOF using saturated vapor cells is also provided. This work shows unique features of the HCL-FADOF in a low-temperature environment and its quick-start advantage, which provides a solid foundation for extensive applications.
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Single-atom-layer catalysts with fully activated basal-atoms will provide a solution to the low loading-density bottleneck of single-atom catalysts. Herein, we activate the majority of the basal sites of monolayer MoS2 , by doping Co ions to induce long-range ferromagnetic order. This strategy, as revealed by inâ situ synchrotron radiation microscopic infrared spectroscopy and electrochemical measurements, could activate more than 50 % of the originally inert basal-plane S atoms in the ferromagnetic monolayer for the hydrogen evolution reaction (HER). Consequently, on a single monolayer of ferromagnetic MoS2 measured by on-chip micro-cell, a current density of 10â mA cm-2 could be achieved at the overpotential of 137â mV, corresponding to a mass activity of 28, 571â Ag-1 , which is two orders of magnitude higher than the multilayer counterpart. Its exchange current density of 75â µA cm-2 also surpasses most other MoS2 -based catalysts. Experimental results and theoretical calculations show the activation of basal plane S atoms arises from an increase of electronic density around the Fermi level, promoting the H adsorption ability of basal-plane S atoms.
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Available safety evaluations regarding mesoporous silica nanoparticles (mSiNPs) are based on the assumption of a relatively high exposure concentration, which makes the findings less valuable in a realistic environment. In this study, we employed Caenorhabditis elegans (C. elegans) as a model to assess the neuronal damage caused by mSiNPs at the predicted environmentally relevant concentrations. After nematodes were acute and prolonged exposed to mSiNPs at concentrations over 300 µg/L, locomotion degeneration, shrinking behavior, and abnormal foraging behavior were observed, which were associated with the deficits in the development of GABAergic neurons, including D-type and RME motor neurons. Furthermore, the oxidative stress evidenced by excessive ROS generation might contribute to the mechanism of mSiNPs damaging neurons. Although the neurotoxicity of mSiNPs was weaker than (nonmesoporous) SiNPs, it is still necessary for researchers to pay attention to the adverse effects caused by mSiNPs in the environmental animals, especially with the rapid increase in mSiNPs application. Considering the conserved property of GABAergic neurons during evolution, these findings will shed light on our understanding of the potential eco-risks of NPs to the nervous system of other animal models.
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Neuronas GABAérgicas/efectos de los fármacos , Neuronas Motoras/efectos de los fármacos , Nanopartículas/química , Dióxido de Silicio/toxicidad , Animales , Caenorhabditis elegans , Relación Dosis-Respuesta a Droga , Neuronas GABAérgicas/metabolismo , Neuronas Motoras/metabolismo , Estrés Oxidativo/efectos de los fármacos , Tamaño de la Partícula , Porosidad , Especies Reactivas de Oxígeno/metabolismo , Dióxido de Silicio/química , Propiedades de SuperficieRESUMEN
In the original publication of the article, the Acknowledgement section was published incompletely. The complete Acknowledgement is given in this Correction.
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OBJECTIVES: To establish genetically modified cell lines that can produce functional α1-antitrypsin (AAT), by CRISPR/Cas9-assisted homologous recombination. RESULTS: α1-Antitrypsin deficiency (AATD) is a monogenic heritable disease that often results in lungs and liver damage. Current augmentation therapy is expensive and in short of supply. To develop a safer and more effective therapeutic strategy for AATD, we integrated the AAT gene (SERPINA1, NG_008290.1) into the AAVS1 locus of human cell line HEK293T and assessed the safety and efficacy of CRISPR/Cas9 on producing potential therapeutic cell lines. Cell clones obtained had the AAT gene integrated at the AAVS1 locus and secreted approx. 0.04 g/l recombinant AAT into the medium. Moreover, the secreted AAT showed an inhibitory activity that is comparable to plasma AAT. CONCLUSIONS: CRISPR/Cas9-mediated engineering of human cells is a promising alternative for generating isogenic cell lines with consistent AAT production. This work sheds new light on the generation of therapeutic liver stem cells for AATD.
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Ingeniería Genética/métodos , Células HEK293/citología , alfa 1-Antitripsina/genética , Sistemas CRISPR-Cas , Técnicas de Cultivo de Célula , Dosificación de Gen , Células HEK293/metabolismo , Humanos , Transfección , alfa 1-Antitripsina/metabolismoRESUMEN
The suppressor of cytokine signaling SOCS1 is a member of the cytokine signaling pathway inhibitor family, which is induced by the IFN-γ induced JAK signaling pathway. The expression of SOCS1 has been found to increase in Crohn's disease (CD) patients, but the role of SOCS1 in intestinal epithelium is unclear. This study was designed to investigate whether SOCS1 has a role in the death of intestinal epithelial cells and intestinal injury. The results showed that the expression of SOCS1 increased in CD patients, and the expression of SOCS1, p-p53 and PUMA increased in the mouse TNBS induced colitis model. Using IFN-γ treated HT-29 cells as an apoptotic model of intestinal epithelial cells in vitro, we confirmed that SOCS1 promoted apoptosis of intestinal epithelial cells by activating p53. In HT-29 cells which were treated with IFN-γ, the interaction between p53 and SOCS1 and phosphorylation of p53 were significantly higher than untreated cells. When knocking SOCS1 down by using SOCS1 siRNA, phosphorylation of p53 and apoptosis of intestinal epithelial cells which was induced by IFN-γ were significantly inhibited. In summary, our findings suggest that SOCS1 may promote apoptosis of intestinal epithelial cells at least partly through mediating p53 signaling.
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Apoptosis , Enfermedad de Crohn/patología , Células Epiteliales/patología , Intestinos/patología , Transducción de Señal , Proteína 1 Supresora de la Señalización de Citocinas/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Animales , Apoptosis/efectos de los fármacos , Colitis , Enfermedad de Crohn/metabolismo , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Femenino , Técnicas de Silenciamiento del Gen , Células HT29 , Humanos , Interferón gamma/farmacología , Ratones Endogámicos BALB C , Transducción de Señal/efectos de los fármacos , Ácido TrinitrobencenosulfónicoRESUMEN
BACKGROUND: Pyruvate kinase M2 (PKM2), a key glycolytic enzyme, is involved in multiple cellular processes including apoptosis. Recently increased fecal PKM2 has been found in Crohn's disease (CD), but little is known regarding its function in the pathophysiology of the disease. AIM: The intestinal expression of PKM2 and its involvement in CD was investigated. METHODS: Pyruvate kinase M2 expression in mucosal biopsies from patients with CD and normal controls was detected by immunohistochemistry. A murine model of colitis induced by trinitrobenzenesulphonic acid (TNBS) was established and expression of PKM2, B cell lymphoma-extra large (Bcl-xl), active caspase-3 as well as cleaved poly (ADP-ribose) polymerase (PARP) was examined for association of PKM2 with intestinal epithelial cell (IEC) apoptosis. Furthermore, we treated human IEC line HT-29 by tumor necrosis factor-α (TNF-α) and used RNA interference to analyze the role of PKM2 in IEC apoptosis. RESULTS: Intestinal expression of PKM2 was higher in patients with CD compared with normal controls mainly locating in IECs. In TNBS-induced colitis, up-regulation of PKM2 was accompanied by the elevated expression of Bcl-xl, active caspase-3, and cleaved PARP. PKM2 was co-localized with active caspase-3 in IECs marked by E-cadherin, suggesting its role in IEC apoptosis. Expression of PKM2 and Bcl-xl in TNF-α-induced HT-29 cells was increased, while TNF-α had no effect on cellular localization of PKM2. Furthermore, knockdown of PKM2 by siRNA could inhibit expression of Bcl-xl but enhance apoptosis in TNF-α-treated HT-29 cells. CONCLUSION: The up-regulation of PKM2 might protect IECs against apoptosis possibly through Bcl-xl in CD, indicating its important role in the pathophysiology of CD.
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Apoptosis , Proteínas Portadoras/metabolismo , Colon/enzimología , Enfermedad de Crohn/enzimología , Células Epiteliales/enzimología , Mucosa Intestinal/enzimología , Proteínas de la Membrana/metabolismo , Piruvato Quinasa/metabolismo , Hormonas Tiroideas/metabolismo , Animales , Proteínas Reguladoras de la Apoptosis/metabolismo , Proteínas Portadoras/genética , Estudios de Casos y Controles , Colon/patología , Enfermedad de Crohn/inducido químicamente , Enfermedad de Crohn/patología , Modelos Animales de Enfermedad , Células Epiteliales/patología , Femenino , Células HT29 , Humanos , Mediadores de Inflamación/metabolismo , Mucosa Intestinal/patología , Proteínas de la Membrana/genética , Ratones , Ratones Endogámicos BALB C , Estudios Prospectivos , Piruvato Quinasa/genética , Interferencia de ARN , Transducción de Señal , Hormonas Tiroideas/genética , Factores de Tiempo , Transfección , Ácido Trinitrobencenosulfónico , Factor de Necrosis Tumoral alfa/metabolismo , Proteínas de Unión a Hormona TiroideRESUMEN
BACKGROUND: Successful viral infection requires the involvement of host cellular factors in their life cycle. Heat shock protein 70 (HSP70) can be recruited by numerous viruses to promote the folding, maturation, or assembly of viral proteins. We have previously shown that HSP70 is significantly elevated in porcine reproductive and respiratory syndrome virus (PRRSV)-infected lungs, suggesting HSP70 may play a potential role during PRRSV infection. In this study, we tried to investigate the role of HSP70 during PRRSV infection. RESULTS: In this study, we observed that PRRSV infection induced the expression of HSP70. The down-regulation of HSP70 using quercetin, a HSPs synthesis inhibitor, or small interfering RNAs (siRNA) reduced the viral protein level and viral production. Notably, these inhibitory effects on PRRSV infection could be attenuated by heat shock treatment. In addition, HSP70 was found to colocalize with the viral double-stranded RNA (dsRNA) and knockdown of HSP70 decreased the dsRNA levels, suggesting HSP70 is involved in the formation of viral replication and transcription complex (RTC) and thus affects the viral replication. CONCLUSIONS: Our study revealed that HSP70 is an essential host factor required for the replication of PRRSV. The inhibition of HSP70 significantly reduced PRRSV replication, which may be applied as an effective antiviral strategy.
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Proteínas HSP70 de Choque Térmico/metabolismo , Interacciones Huésped-Patógeno , Virus del Síndrome Respiratorio y Reproductivo Porcino/fisiología , Replicación Viral , Animales , Técnicas de Silenciamiento del Gen , Proteínas HSP70 de Choque Térmico/antagonistas & inhibidores , Quercetina/metabolismo , ARN Interferente Pequeño/metabolismo , PorcinosRESUMEN
OCT4 is a well-established regulator of pluripotency and nuclear reprogramming. To determine if improving OCT4 abundance can facilitate oocyte-mediated reprogramming in cloned porcine embryos, we artificially increased OCT4 levels by co-incubating donor cells with 50 ng/µl OCT4 plasmid. We observed higher rates of blastocyst formation (P < 0.05) and lower levels of blastocyst apoptosis in nuclear-transfer-derived embryos carrying OCT4-incubated donor nuclei (OCT4-SCNT). The beneficial effect caused by exogenous expression of OCT4 involves epigenetic changes, wherein increased histone acetylation (AcH3K9) appeared in OCT4-SCNT embryos at the one-cell and blastocyst stages and reduced histone methylation (H3K9me2) was observed at the one-cell stage (P < 0.05). There was a transient increase in exogenous OCT4 and an up-regulation of endogenous OCT4 level in OCT4-SCNT embryos (P < 0.05), while the expression pattern of epigenetic enzymes was changed. These modifications were accompanied by an up-regulation of CDX2, whose interaction with OCT4 is instrumental for implantation, and a down-regulation of XIST, a negative indicator of reprogramming (P < 0.05). Taken together, our results support a role for exogenous expression of OCT4 in improving the efficiency of nuclear reprogramming while establishing a convenient and timesaving method to improve nuclear-transfer outcomes.
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Blastocisto/fisiología , Reprogramación Celular/fisiología , Factor 3 de Transcripción de Unión a Octámeros/biosíntesis , Factor 3 de Transcripción de Unión a Octámeros/genética , Plásmidos/administración & dosificación , Animales , Apoptosis/efectos de los fármacos , Blastocisto/metabolismo , Epigénesis Genética/efectos de los fármacos , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Humanos , Técnicas de Transferencia Nuclear , Factor 3 de Transcripción de Unión a Octámeros/metabolismo , Plásmidos/genética , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Alineación de Secuencia , PorcinosRESUMEN
BACKGROUND: Porcine reproductive and respiratory syndrome virus (PRRSV) infection leads to substantial economic losses to the swine industry worldwide. However, no effective countermeasures exist to combat this virus infection so far. The most common antiviral strategy relies on directly inhibiting viral proteins. However, this strategy invariably leads to the emergence of drug resistance due to the error-prone nature of viral ploymerase. Targeting cellular proteins required for viral infection for developing new generation of antivirals is gaining concern. Recently, heat shock protein 90 (HSP90) was found to be an important host factor for the replication of multiple viruses and the inhibition of HSP90 showed significant antiviral effects. It is thought that the inhibition of HSP90 could be a promising broad-range antiviral approach. However, the effects of HSP90 inhibition on PRRSV infection have not been evaluated. In the current research, we tried to inhibit HSP90 and test whether the inhibition affect PRRSV infection. METHODS: We inhibit the function of HSP90 with two inhibitors, geldanamycin (GA) and 17- allylamono-demethoxygeldanamycin (17-AAG), and down-regulated the expression of endogenous HSP90 with specific small-interfering RNAs (siRNAs). Cell viability was measured with alamarBlue. The protein level of viral N was determined by western blotting and indirect immunofluorescence (IFA). Besides, IFA was employed to examine the level of viral double-stranded RNA (dsRNA). The viral RNA copy number and the level of IFN-ß mRNA were determined by quantitative real-time PCR (qRT-PCR). RESULTS: Our results indicated that both HSP90 inhibitors showed strong anti-PRRSV activity. They could reduce viral production by preventing the viral RNA synthesis. These inhibitory effects were not due to the activation of innate interferon response. In addition, we observed that individual knockdown targeting HSP90α or HSP90ß did not show dramatic inhibitory effect. Combined knockdown of these two isoforms was required to reduce viral infection. CONCLUSIONS: Our results shed light on the possibility of developing potential therapeutics targeting HSP90 against PRRSV infection.
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Proteínas HSP90 de Choque Térmico/metabolismo , Interacciones Huésped-Patógeno , Virus del Síndrome Respiratorio y Reproductivo Porcino/fisiología , Replicación Viral , Animales , Benzoquinonas/metabolismo , Western Blotting , Línea Celular , Supervivencia Celular/efectos de los fármacos , Técnica del Anticuerpo Fluorescente Indirecta , Técnicas de Silenciamiento del Gen , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Lactamas Macrocíclicas/metabolismo , ARN Viral/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa , Porcinos , Proteínas Virales/análisisRESUMEN
Xylanase gene xynB, cellulase genes egxA and bgl4 were subjected to codon optimisation using two opposing strategies. One was designated the 'one amino acid-one codon' approach, which employs only the codon most used by humans for each amino acid. The other one is referred to as the "humanised" codon usage method, which selects synonymous codons for each amino acid according to the human codon usage table to mimic patterns used in humans. Protein expression levels in mammalian cell lines from each sequence were measured using fluorescence-activated cell sorting, western blotting and enzymatic activity assay. The results indicate that compared with the humanised codon usage method, the relatively simple 'one amino acid-one codon' approach could enhance heterologous protein expression in mammalian cells without apparent drawbacks.