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In this study, non-thermal plasma (NTP) was employed to modify the Cu/TiO2 adsorbent to efficiently purify H2S in low-temperature and micro-oxygen environments. The effects of Cu loading amounts and atmospheres of NTP treatment on the adsorption-oxidation performance of the adsorbents were investigated. The NTP modification successfully boosted the H2S removal capacity to varying degrees, and the optimized adsorbent treated by air plasma (Cu/TiO2-Air) attained the best H2S breakthrough capacity of 113.29 mg H2S/gadsorbent, which was almost 5 times higher than that of the adsorbent without NTP modification. Further studies demonstrated that the superior performance of Cu/TiO2-Air was attributed to increased mesoporous volume, more exposure of active sites (CuO) and functional groups (amino groups and hydroxyl groups), enhanced Ti-O-Cu interaction, and the favorable ratio of active oxygen species. Additionally, the X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS) results indicated the main reason for the deactivation was the consumption of the active components (CuO) and the agglomeration of reaction products (CuS and SO42-) occupying the active sites on the surface and the inner pores of the adsorbents.

Asunto(s)

Cobre , Sulfuro de Hidrógeno , Oxidación-Reducción , Titanio , Titanio/química , Adsorción , Cobre/química , Sulfuro de Hidrógeno/química , Contaminantes Atmosféricos/química , Gases em Plasma/química , Modelos Químicos

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The direct coupling of nitrate ions and carbon dioxide for urea synthesis presents an appealing alternative to the Bosch-Meiser process in industry. The simultaneous activation of carbon dioxide and nitrate, however, as well as efficient C-N coupling on single active site, poses significant challenges. Here, we propose a novel metal/hydroxide heterostructure strategy based on synthesizing an Ag-CuNi(OH)2 composite to cascade carbon dioxide and nitrate reduction reactions for urea electrosynthesis. The strongly coupled metal/hydroxide heterostructure interface integrates two distinct sites for carbon dioxide and nitrate activation, and facilitates the coupling of *CO (on silver, where * denotes an active site) and *NH2 (on hydroxide) for urea formation. Moreover, the strongly coupled interface optimizes the water splitting process and facilitates the supply of active hydrogen atoms, thereby expediting the deoxyreduction processes essential for urea formation. Consequently, our Ag-CuNi(OH)2 composite delivers a high urea yield rate of 25.6 mmol gcat.-1 h-1 and high urea Faradaic efficiency of 46.1%, as well as excellent cycling stability. This work provides new insights into the design of dual-site catalysts for C-N coupling, considering their role on the interface.

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The spoilage of irradiated ready-to-eat chicken feet (RTECF) seriously affects the food's quality, resulting in package swelling and off-flavors, both of which are highly undesirable to stakeholders and consumers. To investigate the spoilage characteristics of irradiated RTECF and the microorganisms responsible for the spoilage and swelling, the changes in physicochemical properties, microbial community, and volatile organic compounds (VOCs) between normal and spoiled RTECF were evaluated. Compared with normal samples, the spoiled RTECF showed a higher pH value and total volatile basic nitrogen (TVB-N) value, lower color value, and texture features (P < 0.05). Acinetobacter, Pseudomonas, Lactobacillus, and Candida were the dominant genera responsible for RTECF spoilage as confirmed through both culture-dependent methods and high-throughput sequencing (HTS). The results of the verification for gas-producing strains showed that Lactobacillus brevis could cause RTECF packaging to swell. A total of 20 key VOCs were identified using headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS). The results of Pearson correlation analysis (|r|>0.8, P < 0.05) showed that 12 dominant core microbial genera had a significant effect on the flavor of RTECF before and after spoilage. This study provides a theoretical reference for solving the problem of RTECF spoilage and improving the overall quality of RTECF products.

Asunto(s)

Bacterias , Pollos , Irradiación de Alimentos , Microbiología de Alimentos , Compuestos Orgánicos Volátiles , Pollos/microbiología , Animales , Compuestos Orgánicos Volátiles/análisis , Compuestos Orgánicos Volátiles/química , Compuestos Orgánicos Volátiles/metabolismo , Bacterias/clasificación , Bacterias/efectos de la radiación , Bacterias/aislamiento & purificación , Bacterias/genética , Bacterias/crecimiento & desarrollo , Bacterias/metabolismo , Irradiación de Alimentos/métodos , Microbiota/efectos de la radiación , Embalaje de Alimentos/métodos , Cromatografía de Gases y Espectrometría de Masas , Concentración de Iones de Hidrógeno , Comida Rápida/microbiología , Comida Rápida/análisis

RESUMEN

BACKGROUND AND OBJECTIVES: Neonatal cardiac surgery requires careful consideration of cardiopulmonary bypass (CPB) priming fluid composition due to small blood volume and immature physiology. This study investigated the impact of allogeneic stored red blood cells (RBCs) processed using an autotransfusion system in CPB priming fluid for neonates. MATERIALS AND METHODS: We compared perioperative parameters, inflammatory mediators, coagulation indicators, vasoactive-inotropic score (VIS) and clinical outcomes between neonates receiving unwashed (n = 56) and washed (n = 45) RBCs in CPB priming fluid. Regression models were used to assess the independent association between RBC washing and patient outcomes. RESULTS: The autotransfusion system improved stored RBC quality. The washed group showed higher peak haematocrit (p < 0.01) and haemoglobin levels (p = 0.04) during CPB, an increased oxygen delivery index during rewarming (p < 0.05) and lower postoperative lactate levels and VIS (p < 0.05). Inflammatory (IL-6, IL-8 and IL-10) and coagulation parameters (D-dimer, fibrinogen and fibrin degradation product) fluctuated compared with baseline but did not significantly differ between groups. The washed group had a lower incidence of hyperlactacidaemia and delayed sternal closure at CPB weaning. CONCLUSIONS: Adding washed allogeneic stored RBCs to neonatal CPB priming fluid reduced postoperative lactate elevation and VIS without early improvement in the inflammatory and coagulation systems.

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Identification of potential cerebrovascular disorder in the patient with traumatic subarachnoid hemorrhage (tSAH) is a key element to decrease the complication occurrence and mortality rate. In this study, we aim to compare the diagnostic values between dual-source computed tomography angiography (DSCTA) and traditional tomography angiography (CTA) in identification of potential cerebrovascular disorder among tSAH patients. A total of 113 tSAH patients with the hemorrhage involving more than 2 cisterns were recruited. Among that, 42 patients received DSCTA scans, and another 71 patients received traditional CTA scans. Subsequently, all patients received digital subtraction angiography (DSA) tests to confirm the presence of the cerebrovascular disorder. In DSCTA scan group, 21.4 % (9/42) patients were reported to have cerebrovascular disorders: seven patients had intracranial aneurysms; a patient had pseudoaneurysm with carotid artery cavernous sinus fistula; and a patient had Moyamoya disease. DSA tests had the same results with that with DSCTA scans. In the cohort receiving CTA scans, 19.7 % (14/71) patients were reported to had intracranial aneurysms. However, the positive results of DSA tests for this cohort were 22.5 % (16/71). Two inconsistent results between the CTA scan and DSA test were found, including an arteriovenous malformation and an arteriovenous fistula. In summary, DSCTA and CTA had similar positive rates but differ in diagnostic accuracy for identification of cerebrovascular disorders in tSAH patients.

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Computing in memory (CIM) breaks the conventional von Neumann bottleneck through in situ processing. Monolithic integration of digital and analog CIM hardware, ensuring both high precision and energy efficiency, provides a sustainable paradigm for increasingly sophisticated artificial intelligence (AI) applications but remains challenging. Here, we propose a complementary metal-oxide semiconductor-compatible ferroelectric hybrid CIM platform that consists of Boolean logic and triggers for digital processing and multistage cell arrays for analog computation. The basic ferroelectric-gated units are assembled with solution-processable two-dimensional (2D) molybdenum disulfide atomic-thin channels at a wafer-scale yield of 96.36%, delivering high on/off ratios (>107), high endurance (>1012), long retention time (>10 years), and ultralow cycle-to-cycle/device-to-device variations (~0.3%/~0.5%). Last, we customize a highly compact 2D hybrid CIM system for dynamic tracking, achieving a high accuracy of 99.8% and a 263-fold improvement in power efficiency compared to graphics processing units. These results demonstrate the potential of 2D fully ferroelectric-gated hybrid hardware for developing versatile CIM blocks for AI tasks.

RESUMEN

BACKGROUND: Obesity-related hypertension is a major cardiovascular risk factor. Apigenin, a natural flavonoid in celery, induces vascular dilation via endothelial transient receptor potential channel vanilla 4 (TRPV4) channels. This study aimed to explore apigenin's potential to alleviate obesity-related hypertension in mice and its underlying mechanisms. METHODS: The C57BL/6 and TRPV4 knockout mice were fed a high-fat diet and subjected to dietary intervention with apigenin. Body weight and tail blood pressure of the mice were measured during the feeding. Vascular reactivity was assessed through a DMT wire myograph systems in vitro. The distribution and expression of adiponectin and pro-inflammatory markers in brown fat were detected. Injecting adeno-associated eight (AAV8) viruses into brown adipose tissue (BAT) to determine whether adiponectin is indispensable for the therapeutic effect of apigenin. Palmitic acid (PA) was used in mouse brown adipocytes to examine the detailed mechanisms regulating adiponectin secretion. RESULTS: Apigenin improved vasodilation and reduced blood pressure in obese mice, effects partly blocked in TRPV4 knockout. It also reduced weight gain independently of TRPV4. Apigenin increased adiponectin secretion from BAT; knockdown of adiponectin weakened its benefits. Apigenin downregulated Cluster of differentiation 38 (CD38), restoring Nicotinamide adenine dinucleotide+ (NAD+) levels and activating the NAD+/Sirtuin 1 (SIRT1) pathway, enhancing adiponectin expression. CONCLUSIONS: Our study indicates that dietary apigenin is suitable as a nonpharmaceutical intervention for obesity-related hypertension. In mechanism, in addition to improving vascular relaxation through the activation of endothelial TRPV4 channels, apigenin also directly alleviated adipose inflammation and increased adiponectin levels by inhibiting CD38.

RESUMEN

Infertility has gradually become a global health concern, and evidence suggests that exposure to environmental endocrine-disrupting chemicals (EDCs) represent one of the key causes of infertility. Benzo(a)pyrene (BaP) is a typical EDC that is widespread in the environment. Previous studies have detected BaP in human urine, semen, cervical mucus, oocytes and follicular fluid, resulting in reduced fertility and irreversible reproductive damage. However, the mechanisms underlying the effects of gestational BaP exposure on offspring fertility in male mice have not been fully explored. In this study, pregnant mice were administered BaP at doses of 0, 5, 10 and 20 mg/kg/day via gavage from Days 7.5 to 12.5 of gestation. The results revealed that BaP exposure during pregnancy disrupted the structural integrity of testicular tissue, causing a disorganized arrangement of spermatogenic cells, compromised sperm quality, elevated levels of histone modifications and increased apoptosis in the testicular tissue of F1 male mice. Furthermore, oxidative stress was also increased in the testicular tissue of F1 male mice. BaP activated the AhR/ERα signaling pathway, affected H3K4me3 expression and induced apoptosis in testicular tissue. AhR and Cyp1a1 were overexpressed, and the expression of key molecules in the antioxidant pathway, including Keap1 and Nrf2, was reduced. The combined effects of these molecules led to apoptosis in testicular tissues, damaging and compromising sperm quality. This impairment in testicular cells further contributed to compromised testicular tissues, ultimately impacting the reproductive health of F1 male mice.

RESUMEN

Telethonin/titin-cap (TCAP) encodes a Z-disc protein that plays important roles in sarcomere/T-tubule interactions, stretch-sensing and signaling. Mutations in TCAP are associated with muscular dystrophy and cardiomyopathy; however, the complete etiology and its roles in myocardial infarction and regeneration are not fully understood. Here, we generated tcap gene knockout zebrafish with CRISPR/Cas9 technology and observed muscular dystrophy-like phenotypes and abnormal mitochondria in skeletal muscles. The stretch-sensing ability was inhibited in tcap-/- mutants. Moreover, Tcap deficiency led to alterations in cardiac morphology and function as well as increases in reactive oxygen species (ROS) and mitophagy. In addition, the cardiac regeneration and cardiomyocyte proliferation ability of tcap-/- mutants were impaired, but these impairments could be rescued by supplementation with ROS scavengers or autophagy inhibitors. Overall, our study demonstrates the essential roles of Tcap in striated muscle function and heart regeneration. Additionally, elevations in ROS and autophagy may account for the phenotypes resulting from Tcap deficiency and could serve as novel therapeutic targets for muscular dystrophy and cardiomyopathy.

RESUMEN

Microplastics (MPs) and per- and polyfluoroalkyl substances (PFASs) coexist widely in lakes and affect ecological security. The coexistence characteristics and adsorption-desorption mechanisms between MPs and typical PFASs were explored in a typical eutrophic shallow lake (Taihu Lake). Polyvinyl chloride (PVC) and polyethylene (PE) are the primary types of MPs in Taihu Lake, with average abundances in water and sediment of 18630 n/m3 and 584 n/kg, respectively. The average concentrations of PFASs in water and sediment are 288.93 ng/L and 4.33 ng/g, with short-chain PFASs (C4-C7) being the main pollutants. Perfluorobutanoic acid (PFBA) in both water and sediment contributed 38.48 % and 44.53 %, respectively, followed by hexafluoropropylene oxide dimer acid (HFPO-DA). The morphological characteristics of MPs influence the distribution of long-chain PFAS in lake water, while the presence of HFPO-DA and perfluorohexanoic acid (PFHxA) in sediment is directly linked to the concentration and size of MPs. A combination of field investigations and indoor experiments revealed that the irreversible adsorption characteristics between MPs and HFPO-DA may promote the high cumulative flux of HFPO-DA in sediment, and the biofilm on the surface of MPs significantly accelerates this accumulation process. The results provide a new perspective on the co-transport behavior of emerging pollutants in aquatic environments.

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We present a TALEN-based workflow to generate and maintain dual-edited (IL-15+/+/TGFßR2-/-) iPSCs that produce enhanced iPSC-derived natural killer (iNK) cells for cancer immunotherapy. It involves using a cell lineage promoter for knocking in (KI) gene(s) to minimize the potential effects of expression of any exogenous genes on iPSCs. As a proof-of-principle, we KI IL-15 under the endogenous B2M promoter and show that it results in high expression of the sIL-15 in iNK cells but minimal expression in iPSCs. Furthermore, given that it is known that knockout (KO) of TGFßR2 in immune cells can enhance resistance to the suppressive TGF-ß signaling in the tumor microenvironment, we develop a customized medium containing Nodal that can maintain the pluripotency of iPSCs with TGFßR2 KO, enabling banking of these iPSC clones. Ultimately, we show that the dual-edited IL-15+/+/TGFßR2-/- iPSCs can be efficiently differentiated into NK cells that show enhanced autonomous growth and are resistant to the suppressive TGF-ß signaling.

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High-risk human papilloma virus (HR-HPV) persistent infection is closely associated with the development of cervical cancer and squamous intraepithelial lesion (SIL).The α-9 HPVs, which is predominantly composed of HR-HPV types, account for 75% of HR-HPV infection in Sichuan. The oncoproteins E6 and E7 of HPV play a crucial role in tumor initiation and progression. Notably, HPV-35 is the only HR-HPV type within the α-9 genus that is not included in the nine-valent HPV prophylactic vaccine. Cervical cell samples obtained from Sichuan were collected for HPV detection and genotyping. Among the 406 HPV-positive samples, 31 HPV-35 were detected, 24 HPV-35 E6 and 26 E7 were successfully amplified and sequenced, five nucleotide mutations in E6 and three in E7 were detected, T232C, T434G of E6 (W78R, I145R) and C67T, G84T of E7 (H23Y, L28F) were non-synonymy mutation. PAML 4.8 server was used to detect positive selection sites of HPV-35 E6, E7, and E6 is W78R. Phyre2 were used to predict and analyze protein structures, W78R made influences on protein structure. IEDB were used to screen epitopes vaccine target for HPV-35 affection therapy, and 5 HPV-35 E6 and 3 HPV-35 E7 most potential epitopes were obtained, the most potential peptides for therapy vaccine design were 79-91YRYSVYGETLEKQ, 45-60FACYDLCIVREGQPY, 124-135RFHNIGGRWTGR of E6; 3-19GEITTLQDYVLDLEPEA, 38-47TIDGPAGQAK, 70-88VQSTHIDIRKLEDLLMGTF of E7 and W78R mainly decreased the epitopes affinity.Conclusions Amino acid substitution in the positive selection sites of HPV-35 E6 and E7 genes have been found to influence protein structure and to decrease the overall affinity of antigen epitopes. This observation aligns with the evolutionary significance of positive selection site, which may confer advantages to the virus by making infected cells more challenging for the immune system to detect, thereby enhancing HPV's adaptability to the host environment. The polymorphism analysis of HPV-35 E6, E7 contributes to the enrichment of α-9 HPV data in Sichuan China, which is instrumental in improving the effectiveness of clinical detection. Furthermore, these findings provide a relevant theoretical foundation for the prevention and treatment of HPV-related diseases.

Asunto(s)

Proteínas Oncogénicas Virales , Infecciones por Papillomavirus , Vacunas contra Papillomavirus , Humanos , Proteínas Oncogénicas Virales/genética , Proteínas Oncogénicas Virales/inmunología , Femenino , China , Infecciones por Papillomavirus/virología , Infecciones por Papillomavirus/prevención & control , Vacunas contra Papillomavirus/inmunología , Vacunas contra Papillomavirus/genética , Polimorfismo Genético , Proteínas E7 de Papillomavirus/genética , Proteínas E7 de Papillomavirus/inmunología , Genotipo , Adulto , Neoplasias del Cuello Uterino/virología , Neoplasias del Cuello Uterino/prevención & control , Epítopos/inmunología , Epítopos/genética , Alphapapillomavirus/genética , Alphapapillomavirus/inmunología , Alphapapillomavirus/clasificación , Persona de Mediana Edad , Mutación , Virus del Papiloma Humano

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Bacterial-derived micro-/nanomedicine has garnered considerable attention in anticancer therapy, owing to the unique natural features of bacteria, including specific targeting ability, immunogenic benefits, physicochemical modifiability, and biotechnological editability. Besides, bacterial components have also been explored as promising drug delivery vehicles. Harnessing these bacterial features, cutting-edge physicochemical and biotechnologies have been applied to attenuated tumor-targeting bacteria with unique properties or functions for potent and effective cancer treatment, including strategies of gene-editing and genetic circuits. Further, the advent of bacteria-inspired micro-/nanorobots and mimicking artificial systems has furnished fresh perspectives for formulating strategies for developing highly efficient drug delivery systems. Focusing on the unique natural features and advantages of bacteria, this review delves into advances in bacteria-derived drug delivery systems for anticancer treatment in recent years, which has experienced a process from living entities to artificial mimicking systems. Meanwhile, a summary of relative clinical trials is provided and primary challenges impeding their clinical application are discussed. Furthermore, future directions are suggested for bacteria-derived systems to combat cancer.

Asunto(s)

Bacterias , Neoplasias , Humanos , Neoplasias/terapia , Bacterias/genética , Sistemas de Liberación de Medicamentos/métodos , Animales , Materiales Biomiméticos/química , Antineoplásicos/química , Antineoplásicos/uso terapéutico , Antineoplásicos/farmacología , Nanomedicina/métodos

RESUMEN

Consecutive uranium extraction from seawater is a promising approach to secure the long-term supply of uranium and the sustainability of nuclear energy. Here, we report an ultra-highly efficient strategy via studtite nanodots growth with impressive uranyl uptake capacity of ~ 154.50 mg/g from natural seawater in 12 consecutive days (i.e., average for ~ 12.875 mg/g/day). Uranyl can be extracted as studtite under visible light via the reaction between the adsorbed uranyl and the photogenerated H2O2 with imine-based Covalent-Organic Framework photocatalysts. In detail, over Tp-Bpy, Tp-Bpy-2 and Tp-Py with multiple uranyl chelating sites, uranyl is found extracted as studtite nanodots which can be eluted readily, while over Tp-Bd and Tb-Bpy, uranyl is transformed into studtite nanorods that is more inert for elution. Abundant chelating sites of uranyl via structural regulation of COF photocatalysts are proved to facilitate the formation and efficient elution of studtite nanodots.

RESUMEN

O3-type NaNi0.5Mn0.5O2 (NNM) is very competitive for sodium-ion batteries (SIBs) due to its high capacity and easy production. Nevertheless, the intricate phase transitions during the charging-discharging significantly impede its practical application. This paper proposes a strategy for successfully synthesizing NaNi0.5Mn0.3Ti0.2O2 (NNMT) by combining coprecipitation and a high-temperature solid-state method. This method introduces Ti elements while retaining the electrochemically active Ni2+ content, thus, the NNMT has a high initial specific capacity of 151.4 mAh g-1 at 1 C. It is demonstrated that introducing Ti4+ leads to the transition metal layers becoming disordered by ex situ XRD, thus mitigating the irreversible phase transition of the material. In addition, Ti4+ does not have an outer electron, which can reduce electron delocalization in the transition metal layer and improve the material's cyclic stability. The NNMT possesses a capacity retention rate of 60.66% after 150 cycles, much higher than the initial NNM's 18.96%. It also exhibits an excellent discharge capacity of 86.8 mAh g-1 at 5 C. In conclusion, the cycling and rate performance of the Ti-substituted NNMT are greatly improved without capacity loss, which offers innovative concepts for the modification means of the SIBs layered oxide cathode materials.

RESUMEN

Identifying priority pollutants in wastewater treatment plant (WWTP) effluents is crucial for optimizing monitoring efforts, improving regulations, and developing targeted mitigation strategies. Despite the presence of numerous trace organic pollutants in WWTP effluents, a comprehensive prioritization scheme is lacking, hindering effective control. This study screened 216 micropollutants, including pharmaceuticals, pesticides, and industrial chemicals, which had been detected in effluents from 46 WWTPs across China. A multi-criteria prioritization method was developed, considering exposure potential based on median concentrations and detection frequencies, as well as hazard potential determined by persistence, bioaccumulation, in vitro toxicity, and in vivo toxicity. Pollutants with low exposure or hazard potential were filtered out, and a priority index was calculated to rank the remaining 59 substances. The top 15 priority pollutants included regulated persistent organic pollutants like perfluorooctanoic acid and their alternatives such as perfluorobutane sulfonate, pesticide transformation products, and emerging contaminants such as bisphenol A, which are not currently regulated in WWTP effluents. This study provides a systematic approach to identify priority pollutants and generates a guiding framework for monitoring, regulation, and control of both well-recognized and overlooked contaminants in WWTP effluents.

Asunto(s)

Monitoreo del Ambiente , Aguas Residuales , Contaminantes Químicos del Agua , China , Contaminantes Químicos del Agua/análisis , Aguas Residuales/química , Eliminación de Residuos Líquidos , Ecosistema , Contaminantes Orgánicos Persistentes

RESUMEN

The 5' untranslated region (5'UTR) of many positive-stranded RNA viruses contain functional regulatory sequences. Here, we show that the porcine reproductive and respiratory syndrome virus (PRRSV), a member of arteriviruses, harbors small upstream open reading frames (uORFs) in its 5'UTR. Bioinformatics analysis shows that this feature is relatively well conserved among PRRSV strains and Arteriviridae. We also identified a uORF, namely uORF2, in the PRRSV strain JXwn06, that possesses translational activity and exerts a suppressive effect on the expression of the primary ORF evidenced by in vitro reporter assays. We tested its importance via reverse genetics by introducing a point mutation into the PRRSV infectious cDNA clone to inactivate the start codon of uORF2. The recovered mutant virus Mut2 surprisingly replicated to the same level as the wild-type virus (WT), but induced a higher level of inflammatory cytokines (e.g., TNF-α, IL-1ß, and IL-6) both in vitro and in animal experiments, correlating well with more severe lung injury and higher death rate. In line with this, over-expression of uORF2 in transfected cells significantly inhibited poly(I:C)-induced expression of inflammatory cytokines. Together, our data support the idea that uORF2 encodes a novel, functional regulator of PRRSV virulence despite of its short size. IMPORTANCE: PRRSV has remained a major challenge to the world swine industry, but we still do not know much about its biology and pathogenesis. Here, we provide evidence to show that the 5'UTR of PRRSV strain JXwn06 harbors a functional uORF that has the coding capacity and regulates induction of inflammation as demonstrated by in vitro assays and animal experiment. The findings reveal a novel viral factor that regulates cellular inflammation and provide insight into the understanding of PRRSV pathogenesis.

Asunto(s)

Regiones no Traducidas 5' , Sistemas de Lectura Abierta , Síndrome Respiratorio y de la Reproducción Porcina , Virus del Síndrome Respiratorio y Reproductivo Porcino , Virus del Síndrome Respiratorio y Reproductivo Porcino/genética , Virus del Síndrome Respiratorio y Reproductivo Porcino/fisiología , Animales , Regiones no Traducidas 5'/genética , Porcinos , Síndrome Respiratorio y de la Reproducción Porcina/virología , Replicación Viral , Inflamación/virología , Línea Celular , Citocinas/metabolismo , Citocinas/genética

RESUMEN

Porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV) and transmissible gastroenteritis virus (TGEV) are three clinically common coronaviruses causing diarrhea in pigs, with indistinguishable clinical signs and pathological changes. Rapid, portable and reliable differential diagnosis of these three pathogens is crucial for the prompt implementation of appropriate control measures. In this study, we developed a triplex nucleic acid assay that combines reverse transcription recombinase-aided amplification (RT-RAA) with lateral flow assay (LFA) by targeting the most conserved genomic region in the ORF1b genes of PEDV, PDCoV and TGEV. The entire detection process of the triplex RT-RAA-LFA assay included 10-min nucleic acid amplification at 42 °C and 5-min visual LFA readout at room temperature. The assay could specifically differentiate PEDV, PDCoV and TGEV without cross-reaction with any other major swine pathogens. Sensitivity analysis showed that the triplex RT-RAA-LFA assay was able to detect the viral RNA extracted from the spiked fecal samples with the minimum of 1 × 100 TCID50 PEDV, 1 × 104 TCID50 PDCoV, and 1 × 102 TCID50 TGEV per reaction, respectively. Further analysis showed that the 95 % detection limit (LOD) of triplex RT-RAA-LFA for PEDV, PDCoV, and TGEV were 22, 478, and 205 copies of recombinant plasmids per reaction, respectively. The diagnostic performance of triplex RT-RAA-LFA was compared with that of PEDV, PDCoV and TGEV respective commercial real-time RT-PCR kits by testing 114 clinical rectal swab samples in parallel. The total diagnostic coincidence rates of triplex RT-RAA-LFA with real-time RT-PCR kits of PEDV, PDCoV and TGEV were 100 %, 99.1 % and 99.1 %, respectively, and their Kappa values were 1.00, 0.958 and 0.936, respectively. Collectively, the RT-RAA-LFA assay is a powerful tool for the rapid, portable, visual, and synchronous differential diagnosis of PEDV, PDCoV, and TGEV.

Asunto(s)

Infecciones por Coronavirus , Deltacoronavirus , Heces , Técnicas de Amplificación de Ácido Nucleico , Virus de la Diarrea Epidémica Porcina , ARN Viral , Sensibilidad y Especificidad , Enfermedades de los Porcinos , Virus de la Gastroenteritis Transmisible , Animales , Porcinos , Virus de la Gastroenteritis Transmisible/aislamiento & purificación , Virus de la Gastroenteritis Transmisible/genética , Virus de la Diarrea Epidémica Porcina/aislamiento & purificación , Virus de la Diarrea Epidémica Porcina/genética , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/virología , ARN Viral/genética , ARN Viral/aislamiento & purificación , Heces/virología , Infecciones por Coronavirus/diagnóstico , Infecciones por Coronavirus/veterinaria , Infecciones por Coronavirus/virología , Diagnóstico Diferencial , Deltacoronavirus/aislamiento & purificación , Deltacoronavirus/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Técnicas de Amplificación de Ácido Nucleico/veterinaria , Gastroenteritis Porcina Transmisible/diagnóstico , Gastroenteritis Porcina Transmisible/virología , Técnicas de Diagnóstico Molecular/métodos , Diarrea/virología , Diarrea/veterinaria , Diarrea/diagnóstico

RESUMEN

Circular RNAs (circRNAs) have emerged as a promising alternative to linear mRNA, owing to their unique properties and potential therapeutic applications, driving the development of novel approaches for their production. This study introduces a cis-splicing system that efficiently produces circRNAs by incorporating a ribozyme core at one end of the precursor, thereby eliminating the need for additional spacer elements between the ribozyme and the gene of interest (GOI). In this cis-splicing system, sequences resembling homologous arms at both ends of the precursor are crucial for forming the P9.0 duplex, which in turn facilitates effective self-splicing and circularization. We demonstrate that the precise recognition of the second transesterification site depends more on the structural characteristics of P9.0 adjacent to the ωG position than on the nucleotide composition of the P9.0-ωG itself. Further optimization of structural elements, like P10 and P1-ex, significantly improves circularization efficiency. The circRNAs generated through the cis-splicing system exhibit prolonged protein expression and minimal activation of the innate immune response. This study provides a comprehensive exploration of circRNA generation via a novel strategy and offers valuable insights into the structural engineering of RNA, paving the way for future advancements in circRNA-based applications.