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26 pages, 2170 KiB  
Review
Advancements in Bacteriophages for the Fire Blight Pathogen Erwinia amylovora
by Dufang Ke, Jinyan Luo, Pengfei Liu, Linfei Shou, Munazza Ijaz, Temoor Ahmed, Muhammad Shafiq Shahid, Qianli An, Ivan Mustać, Gabrijel Ondrasek, Yanli Wang, Bin Li and Binggan Lou
Viruses 2024, 16(10), 1619; https://doi.org/10.3390/v16101619 (registering DOI) - 16 Oct 2024
Viewed by 406
Abstract
Erwinia amylovora, the causative agent of fire blight, causes significant economic losses for farmers worldwide by inflicting severe damage to the production and quality of plants in the Rosaceae family. Historically, fire blight control has primarily relied on the application of copper [...] Read more.
Erwinia amylovora, the causative agent of fire blight, causes significant economic losses for farmers worldwide by inflicting severe damage to the production and quality of plants in the Rosaceae family. Historically, fire blight control has primarily relied on the application of copper compounds and antibiotics, such as streptomycin. However, the emergence of antibiotic-resistant strains and growing environmental concerns have highlighted the need for alternative control methods. Recently, there has been a growing interest in adopting bacteriophages (phages) as a biological control strategy. Phages have demonstrated efficacy against the bacterial plant pathogen E. amylovora, including strains that have developed antibiotic resistance. The advantages of phage therapy includes its minimal impact on microbial community equilibrium, the lack of a detrimental impact on plants and beneficial microorganisms, and its capacity to eradicate drug-resistant bacteria. This review addresses recent advances in the isolation and characterization of E. amylovora phages, including their morphology, host range, lysis exertion, genomic characterization, and lysis mechanisms. Furthermore, this review evaluates the environmental tolerance of E. amylovora phages. Despite their potential, E. amylovora phages face certain challenges in practical applications, including stability issues and the risk of lysogenic conversion. This comprehensive review examines the latest developments in the application of phages for controlling fire blight and highlights the potential of E. amylovora phages in plant protection strategies. Full article
(This article belongs to the Section Bacterial Viruses)
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<p>Distribution of phages from various families infecting <span class="html-italic">E. amylovora</span> and <span class="html-italic">E. pyrifoliae</span> in different countries. The stacked bar chart was constructed utilizing Chiplot (<a href="https://www.chiplot.online/" target="_blank">https://www.chiplot.online/</a>; accessed on 22 August 2024).</p>
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<p>This phylogenetic tree of <span class="html-italic">E. amylovora</span> phages was constructed with MEGA 7.0 software by using the maximum composite likelihood method based on the terminase large subunit available in published articles. Nodes show the result of 500 bootstrap replicates.</p>
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<p>The figure illustrates an integrated control strategy based on a phage–carrier system and phage cocktail approach. (<b>A</b>) Different phages binding to vector bacteria; (<b>B</b>) phage infestation of vectors; and (<b>C</b>) phage release. The strategy employs a combination of three key elements: phages, UV protectants, and an auxiliary formulation.</p>
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19 pages, 8261 KiB  
Article
The Critical Role of Phenylpropanoid Biosynthesis Pathway in Lily Resistance Against Gray Mold
by Qi Cui, Xinran Li, Shanshan Hu, Dongfeng Yang, Ann Abozeid, Zongqi Yang, Junhao Jiang, Ziming Ren, Danqing Li, Dongze Li, Liqun Zheng and Anhua Qin
Int. J. Mol. Sci. 2024, 25(20), 11068; https://doi.org/10.3390/ijms252011068 (registering DOI) - 15 Oct 2024
Viewed by 270
Abstract
Gray mold caused by Botrytis elliptica is one of the most determinative factors of lily growth and has become a major threat to lily productivity. However, the nature of the lily B. elliptica interaction remains largely unknown. Here, comparative transcriptomic and metabolomic were [...] Read more.
Gray mold caused by Botrytis elliptica is one of the most determinative factors of lily growth and has become a major threat to lily productivity. However, the nature of the lily B. elliptica interaction remains largely unknown. Here, comparative transcriptomic and metabolomic were used to investigate the defense responses of resistant (‘Sorbonne’) and susceptible (‘Tresor’) lily cultivars to B. elliptica infection at 24 hpi. In total, 1326 metabolites were identified in ‘Sorbonne’ and ‘Tresor’ after infection, including a large number of phenylpropanoids. Specifically, the accumulation of four phenylpropanes, including eriodictyol, hesperetin, ferulic acid, and sinapyl alcohol, was significantly upregulated in the B. elliptica-infected ‘Sorbonne’ compared with the infected ‘Tresor’, and these phenylpropanes could significantly inhibit B. elliptica growth. At the transcript level, higher expression levels of F3′M, COMT, and CAD led to a higher content of resistance-related phenylpropanes (eriodictyol, ferulic acid, and sinapyl alcohol) in ‘Sorbonne’ following B. elliptica infection. It can be assumed that these phenylpropanes cause the resistance difference between ‘Sorbonne’ and ‘Tresor’, and could be the potential marker metabolites for gray mold resistance in the lily. Further transcriptional regulatory network analysis suggested that members of the AP2/ERF, WRKY, Trihelix, and MADS-M-type families positively regulated the biosynthesis of resistance-related phenylpropanes. Additionally, the expression patterns of genes involved in phenylpropanoid biosynthesis were confirmed using qRT-PCR. Therefore, we speculate that the degree of gray mold resistance in the lily is closely related to the contents of phenylpropanes and the transcript levels of the genes in the phenylpropanoid biosynthesis pathway. Our results not only improve our understanding of the lily’s resistance mechanisms against B. elliptica, but also facilitate the genetic improvement of lily cultivars with gray mold resistance. Full article
(This article belongs to the Section Molecular Plant Sciences)
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<p>(<b>A</b>) Oriental hybrid ‘Sorbonne’ and Asiatic hybrid ‘Tresor’. (<b>B</b>) Leaf phenotype of ‘Sorbonne’ and ‘Tresor’ after infection with <span class="html-italic">B. elliptica</span>, scale bar = 1 cm. (<b>C</b>) Leaf lesion size over time of ‘Sorbonne’ and ‘Tresor’ after infection with <span class="html-italic">B. elliptica</span>, n = 12. The lesion areas are shown as the means of the three biological replicates ± SD. Asterisks indicate statistically significant differences between resistant and susceptible cultivars according to Student’s <span class="html-italic">t</span>-test (** <span class="html-italic">p</span> &lt; 0.01).</p>
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<p>(<b>A</b>) Identification and functional enrichment analysis of DEGs in lily. (<b>A</b>) Volcanic plots of DEGs. Red and blue points represent the up-regulated and down-regulated DEGs, respectively. RD and SD represent <span class="html-italic">B. elliptica</span>-inoculated samples of ‘Sorbonne’ and ‘Tresor’, respectively. RH and SH represent the controls of ‘Sorbonne’ and ‘Tresor’, respectively. (<b>B</b>) GO enrichment analysis of the DEGs detected in <span class="html-italic">B. elliptica</span>-infected ‘Sorbonne’ compared with its control. Red texts represent four defense-related terms that were specifically enriched in ‘Sorbonne’ after infection relative to ‘Tresor’. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01. (<b>C</b>) The ridge plot of KEGG enrichment analysis for the DEGs. The average log<sub>2</sub>FoldChange value of each gene in the pathway is shown in the ridge plot. If the value is greater than 0, the gene is the upregulated (right), and the gene is downregulated (left) if its value is less than 0. Three biological replicates were used for transcriptomic analysis.</p>
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<p>Classification of metabolites and identification of DAMs in ‘Sorbonne’ and ‘Tresor’ after infection with <span class="html-italic">B. elliptica</span>. (<b>A</b>) Classification and composition of identified metabolites. (<b>B</b>) Principal component analysis (PCA) of the samples based on the identified metabolites. (<b>C</b>) Numbers of DAMs identified in different pairwise comparisons. (<b>D</b>) Venn diagram of DAMs identified in ‘Sorbonne’ and ‘Tresor’ after infection with <span class="html-italic">B. elliptica</span>. Six biological replicates were used for metabolomic analysis.</p>
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<p>KEGG enrichment analysis of DAMs and in-depth analysis of key metabolites. (<b>A</b>) Six DAMs (a–f) with opposite accumulation patterns in ‘Sorbonne’ and ‘Tresor’ after infection with <span class="html-italic">B. elliptica</span> and three DAMs (g–i) were particularly accumulated in ‘Sorbonne’ or ‘Tresor’ after infection with <span class="html-italic">B. elliptica</span>. The horizontal axis indicates the value of log<sub>2</sub>FoldChange. Orange bars represent up-regulated DAMs, and blue bars represent the down-regulated DAMs. (<b>B</b>) KEGG enrichment analysis for DAMs.</p>
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<p>Integrated analysis of transcriptome and metabolome. (<b>A</b>) Venn diagram of KEGG pathways enriched in DEGs and DAMs in ‘Sorbonne’ and ‘Tresor’. MRD and MSD represent <span class="html-italic">B. elliptica</span>-inoculated samples of ‘Sorbonne’ and ‘Tresor’ metabolomes, respectively. MRH and MSH represent the controls of ‘Sorbonne’ and ‘Tresor’ metabolomes, respectively. TRD and TSD represent <span class="html-italic">B. elliptica</span>-inoculated samples of ‘Sorbonne’ and ‘Tresor’ transcriptomes, respectively. TRH and TSH represent the controls of ‘Sorbonne’ and ‘Tresor’ transcriptomes, respectively. (<b>B</b>) Diagram of partial phenylpropanoid biosynthesis pathway. The values of log<sub>2</sub>FoldChange for key DEGs are marked. (<b>C</b>) Comparison of the metabolite relative contents (represented as peak areas) between control and <span class="html-italic">B. elliptica</span>-infected ‘Sorbonne’ and ‘Tresor’. Data are presented as the means of the three biological replicates ± SD. Different letters above the bars indicate significant differences between treatments or genotypes according to Duncan’s multiple range test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Transcription regulatory network analysis of phenylpropanes in lily after infection with <span class="html-italic">B. elliptica</span>. (<b>A</b>) Specifically expressed DETFs in ‘Sorbonne’ or ‘Tresor’. The values of log<sub>2</sub>FoldChange for DETFs are marked. (<b>B</b>) Transcription regulator network of phenylpropanes according to integrated analysis of transcriptome and metabolome. Red and orange circles represent the DETFs and DAMs, respectively.</p>
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<p>qRT-PCR validation of key candidate DEGs associated with phenylpropanoids metabolism in lily. The relative expression levels are shown as the means of the three biological replicates ± SD. RD and SD represent <span class="html-italic">B. elliptica</span>-inoculated transcriptomic samples of ‘Sorbonne’ and ‘Tresor’, respectively. RH and SH represent the controls of ‘Sorbonne’ and ‘Tresor’, respectively.</p>
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<p>Inhibition effect of the phenylpropanes on <span class="html-italic">B. elliptica</span> growth. (<b>A</b>) Hesperetin, sinapyl alcohol, eriodictyol, and ferulic acid inhibited the growth of <span class="html-italic">B. elliptica</span>. DMSO was used as the solvent control. Scale bar = 1 cm. (<b>B</b>) The inhibition area of <span class="html-italic">B. elliptica</span> after treatment with hesperetin, sinapyl alcohol, eriodictyol, and ferulic acid, respectively. The inhibition areas are shown as the means of the three biological replicates ± SD. Different letters above the bars indicate significant differences among various treatments according to Duncan’s multiple range test (<span class="html-italic">p</span> &lt; 0.01).</p>
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12 pages, 1075 KiB  
Article
Study on Innovative Laminated Flooring with Resin-Impregnated Paper
by Kankan Zhou, Jianhui Cheng and Minliang Fan
Buildings 2024, 14(10), 3250; https://doi.org/10.3390/buildings14103250 - 14 Oct 2024
Viewed by 301
Abstract
A new type of laminated flooring decorated by resin impregnated paper (LWFWRIP) was designed, with the advantages of low formaldehyde emission, improved flame retardant, and high wear resistance. The structure of this new type of wood flooring is based on the ordinary laminated [...] Read more.
A new type of laminated flooring decorated by resin impregnated paper (LWFWRIP) was designed, with the advantages of low formaldehyde emission, improved flame retardant, and high wear resistance. The structure of this new type of wood flooring is based on the ordinary laminated flooring, followed by a decorative layer of thin wood pieces, and then the transparent improved flame retardant, wear-resistant paper is added to the top. It is found that the hot-pressing temperature is the most significant factor affecting the adhesion of resin impregnated paper. The optimal hot-pressing parameters are selected as the hot-pressing pressure of 3.5 MPa, hot-pressing temperature of 180 °C, and hot-pressing time of 40 s. The new laminated flooring was improved with high flame retardant, high wear-resistant, combined with the conventional advantages of both solid wood composite flooring and reinforced wood flooring. The new laminated flooring decorated by resin impregnated paper has broad application prospects. Full article
(This article belongs to the Section Building Materials, and Repair & Renovation)
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<p>Structure of LWFNRIP (<b>a</b>) and LWFWRIP (<b>b</b>).</p>
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<p>Effect of flame-retardant ingredient addition on viscosity.</p>
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<p>Influence of the proportion of flame-retardant components on flame retardant performance.</p>
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<p>Heat release rate curves of flooring samples.</p>
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25 pages, 1901 KiB  
Article
Economic and Environmental Effects of Farmers’ Green Production Behaviors: Evidence from Major Rice-Producing Areas in Jiangxi Province, China
by Mengling Zhang, Li Zhou, Yuhan Zhang and Wangyue Zhou
Land 2024, 13(10), 1668; https://doi.org/10.3390/land13101668 - 13 Oct 2024
Viewed by 448
Abstract
This study examines the economic and environmental impacts of green production practices among farmers. It aims to contribute to sustainable agricultural development, mitigate agricultural non-point source (NPS) pollution, and align environmental protection with economic growth. This paper utilizes survey data from 1345 farm [...] Read more.
This study examines the economic and environmental impacts of green production practices among farmers. It aims to contribute to sustainable agricultural development, mitigate agricultural non-point source (NPS) pollution, and align environmental protection with economic growth. This paper utilizes survey data from 1345 farm households in the main rice production areas of Jiangxi Province, China, using the example of reduced fertilizer application (RFA) among rice farmers. This study constructs a slack-based measure data envelopment analysis (DEA—SBM) model with undesirable outputs to measure environmental effects and applies an endogenous switching regression model (ESRM) to test the economic and environmental effects of farmers’ adoption of green production technologies. We found the following: (1) The RFA behavior of farmers has a significant positive impact on their net profit per hectare (NPH), helping farmers increase their income, with the increase ranging from 2.05% to 6.54%. (2) Farmers’ RFA behavior has a significant positive impact on agricultural green productivity (AGP), contributing to the improvement of the environment, ranging from 44.09% to 45.35%. (3) A heterogeneity analysis found inconsistencies in the income-enhancing and environmental-enhancing effects at different quantiles of NPH and AGP. Therefore, attention should be placed on improving the agricultural product quality supervision system under the market circulation mechanism, creating land scale conditions conducive to the promotion and application of fertilizer reduction technologies and promoting the implementation of externality internalization compensation systems. Full article
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<p>A framework of the effects of farmers’ RFA.</p>
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<p>The distribution of sample counties. The map data in <a href="#land-13-01668-f001" class="html-fig">Figure 1</a> are from DataV. GeoAtlas. <a href="https://datav.aliyun.com/portal/school/atlas/area_selector" target="_blank">https://datav.aliyun.com/portal/school/atlas/area_selector</a>, accessed on 27 May 2024.</p>
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<p>Probability density of farmers’ NPH in two scenarios.</p>
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<p>Probability density of AGP in two scenarios.</p>
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<p>Interquartile regression coefficients and trends.</p>
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12 pages, 5759 KiB  
Article
Effect of Plow Pan on the Redistribution Dynamics of Water and Nutrient Transport in Soils
by Meng’en Zhang, Shuting Dai, Saima Gul, Lizhi He, Hanbo Chen and Dan Liu
Sustainability 2024, 16(20), 8859; https://doi.org/10.3390/su16208859 - 13 Oct 2024
Viewed by 632
Abstract
Plow pans are an essential part of the agricultural soil structure. By adjusting the soil bulk density and plow pan height, the water and nutrient transport are dynamically redistributed. Plow pans play a crucial role in promoting crop growth, increasing yields, and supporting [...] Read more.
Plow pans are an essential part of the agricultural soil structure. By adjusting the soil bulk density and plow pan height, the water and nutrient transport are dynamically redistributed. Plow pans play a crucial role in promoting crop growth, increasing yields, and supporting sustainable land management. In this study, a column experiment was conducted to investigate the effects of plow pan height (10 cm and 15 cm) and bulk density (1.2, 1.4, and 1.6 g cm⁻3) on soil nutrient and water leaching under high-volume (HV) and low-volume (LV) fertilizer applications. The results reveal that the leachate volume decreased by 61.9% at a plow pan height of 10 cm and by 96.2% at a plow pan height of 15 cm when the bulk density was increased from 1.2 to 1.4 g cm⁻3 under HV conditions. There was no leachate when the plow pan bulk density was 1.6 g cm−3. The reserved concentration of alkali-hydrolyzable N in the plow pan soils was the highest when the plow pan had a bulk density of 1.4 g cm−3 and a height of 15 cm. However, when the plow pan height was 15 cm, the available P content in the plow pan soils decreased by 27.0% and 21.0% at bulk densities of 1.4 g cm⁻3 and 1.6 g cm⁻3, respectively, when compared with 1.2 g cm⁻3. Furthermore, the available P concentrations in the plow pan and subsoil layers decreased with an increase in the plow pan height. The available K concentrations in the topsoil decreased by 26.8% and 24.0% when the plow pan bulk density was increased from 1.2 to 1.4 g cm−3 at heights of 10 and 15 cm, respectively. Thus, the optimal plow pan height and bulk density are closely related to the types of soil nutrients. However, it is clear that excessively high bulk densities (e.g., 1.6 g cm−3) negatively impact soil properties. For different nutrient requirements, a bulk density of 1.2 or 1.4 g cm−3 can be chosen, with each providing suitable options based on the specific nutrient needs. This research offers practical insights into changes in nutrient adsorption and fixation in agricultural production associated with alterations in plow pan bulk density. Full article
(This article belongs to the Special Issue Advancing Water and Soil Sustainability with Integrative Approaches)
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<p>Schematic diagram of soil column leaching experiment.</p>
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<p>Accumulated leachate volumes over time in different treatments. Note: LV and HV represent the fertilizer solution volumes; (<b>A</b>,<b>C</b>) represent the accumulated leachate volumes at a bulk density of 1.2 g cm<sup>−3</sup> when the height of the plow pan was 10 and 15 cm, respectively; (<b>B</b>,<b>D</b>) represent the accumulated leachate volumes at a bulk density of 1.4 g cm<sup>−3</sup> when the height of the plow pan was 10 and 15 cm, respectively. There was no leachate in the T3 and T6 treatments.</p>
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<p>(<b>A</b>) TN, (<b>B</b>) TP, and (<b>C</b>) TK contents in the leachate of different treatments. Note: Different letters above the error bars indicate significant differences between 5 cm and 10 cm aqueous solutions under different treatments (<span class="html-italic">p</span> &lt; 0.05). n and * indicate no significant difference and a significant difference between LV and HV in each treatment at <span class="html-italic">p</span> &lt; 0.05, respectively.</p>
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<p>Alkali-hydrolyzable N concentrations in different soil layers. Note: different letters above the error bars indicate significant difference among treatments in the same soil layer (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Available P concentrations in different soil layers. Note: different letters above the error bars indicate significant differences in the same soil layer (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Available K concentrations in different soil layers after various treatments. Note: different letters above the error bars in each soil layer indicate significant differences among treatments (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Redundancy analysis of nutrients in plow pan and leachate. Note: FV: volume of fertilizer solution; P-B: plow pan soil bulk density; H: plow pan height; LV: leachate volume; TN: total N concentrations in leachate; TP: total P concentrations in leachate; TK: total K concentrations in leachate; AN: alkali-hydrolyzed N concentrations in soil; AP: available P concentrations in soil; AK: available K concentrations in soil.</p>
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19 pages, 11711 KiB  
Article
Long-Term Halocarbon Observations in an Urban Area of the YRD Region, China: Characteristic, Sources Apportionment and Health Risk Assessment
by Yuchun Jiang, Anqi Zhang, Qiaoli Zou, Lu Zhang, Hanfei Zuo, Jinmei Ding, Zhanshan Wang, Zhigang Li, Lingling Jin, Da Xu, Xin Sun, Wenlong Zhao, Bingye Xu and Xiaoqian Li
Toxics 2024, 12(10), 738; https://doi.org/10.3390/toxics12100738 - 12 Oct 2024
Viewed by 324
Abstract
To observe the long-term variations in halocarbons in the Yangtze River Delta (YRD) region, this study analyzes halocarbon concentrations and composition characteristics in Shanxi from 2018 to 2020, exploring their origins and the health effects. The total concentration of halocarbons has shown an [...] Read more.
To observe the long-term variations in halocarbons in the Yangtze River Delta (YRD) region, this study analyzes halocarbon concentrations and composition characteristics in Shanxi from 2018 to 2020, exploring their origins and the health effects. The total concentration of halocarbons has shown an overall increasing trend, which is driven by both regulated substances (CFC-11 and CFC-113) and unregulated substances, such as dichloromethane, chloromethane and chloroform. The results of the study also reveal that dichloromethane (1.194 ± 1.003 to 1.424 ± 1.004 ppbv) and chloromethane (0.205 ± 0.185 to 0.666 ± 0.323 ppbv) are the predominant halocarbons in Shanxi, influenced by local and northwestern emissions. Next, this study identifies that neighboring cities in Zhejiang Province and other YRD areas are potentially affected by backward trajectory models. Notably, chloroform and 1,2-dichloroethane have consistently surpassed acceptable thresholds, indicating a significant carcinogenic risk associated with solvent usage. This research sheds light on the evolution of halocarbons in the YRD region, offering valuable data for the control and reduction in halocarbon emissions. Full article
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<p>Geographic location of the Shanxi site.</p>
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<p>Wind rose diagrams illustrating the distribution of wind direction and frequency for the period 2018-2022 at the Shanxi site.</p>
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<p>Time series of halocarbons at the Shanxi monitoring site from 2018 to 2022, showing (<b>a</b>) concentration levels and (<b>b</b>) chemical species composition.</p>
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<p>Seasonal variation in halocarbons at the Shanxi site.</p>
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<p>Diurnal variation of major halocarbon species and total halocarbons.</p>
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<p>Main potential source regions of halocarbons. (<b>a</b>) represents the total halocarbons, and (<b>b</b>) focuses on a specific substance, as indicated in the lower right corner.</p>
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<p>Possible sources of tracer halocarbons for air pollution transport during 2018–2019 at the Shanxi site.</p>
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<p>Interannual variation in non-carcinogenic risk of atmospheric halocarbons in Shanxi. The dashed line indicates the threshold value.</p>
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<p>Interannual variation in carcinogenic risk of atmospheric halocarbons in Shanxi. The dashed lines indicate the acceptable risk level (1 × 10<sup>−6</sup>) and the tolerable risk level (1 × 10<sup>−4</sup>).</p>
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13 pages, 4324 KiB  
Review
The Recent Research Progress of the Tumor mRNA Vaccine
by Hao Zhao, Miying Li, Jiaren Zhou, Lidan Hu, Shaohong Lu and Pan Li
Vaccines 2024, 12(10), 1167; https://doi.org/10.3390/vaccines12101167 - 12 Oct 2024
Viewed by 884
Abstract
Tumors have long posed a significant threat to human life and health, and the messenger ribonucleic acid (mRNA) vaccine is seen as an attractive approach for cancer immunotherapy due to its developmental simplicity, rapid manufacture, and increased immune safety and efficiency. In this [...] Read more.
Tumors have long posed a significant threat to human life and health, and the messenger ribonucleic acid (mRNA) vaccine is seen as an attractive approach for cancer immunotherapy due to its developmental simplicity, rapid manufacture, and increased immune safety and efficiency. In this review, we have summarized details of the developmental history of mRNA vaccines, discussed the basic molecular structure and the effect on the stable and translation level of mRNA, analyzed the underlying immune efficiency and mechanisms on tumors, and assessed the current status of clinical research. We explored the treatment and application prospects of mRNA vaccines, aiming to provide perspectives on the future of mRNA tumor vaccines for ongoing clinical research. Full article
(This article belongs to the Special Issue mRNA Vaccines: Pioneering the Future of Vaccination)
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<p>Time axis of mRNA vaccine development: summary of the time axis of mRNA vaccine development from 1990 to 2023. pDNA: Plasmid DNA; CNE: Cationic Nanoparticle Emulsion; LNP: Lipid Nanoparticle; DOTAP: N-1-(2,3-diethoxy) propyl-n, n, n-trimethylammonium sulfate; DC: Dendritic cells.</p>
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<p>Diagram of the structures of three different types of mRNA vaccine. This section introduces the fundamental structures of three distinct types of mRNA and the various roles played by their components. By modifying and optimizing specific key elements, mRNA can enhance its efficiency and stability during the translation process. UTR: Untranslated Region, uORF: upstream open reading frame, saRNA: self-amplifying mRNA, circRNA: circular mRNA, CDS: coding DNA sequence, GOI: gene of interest, NSP: non-structural protein.</p>
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<p>Chemical structure of 5′-cap of mRNA. In this specific RNA molecule, the methylation patterns presented by the initial few 5′ nucleotides are observed, and distinct cap variants such as CAP 0, CAP 1, CAP 2, and so on, can be clearly differentiated. The determination of these cap variants depends on the differences in the methylation patterns (as shown in the figure). B1, B2: nucleobases.</p>
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<p>The immune mechanisms of mRNA tumor vaccines in vivo. The mRNA vaccine consisting of mRNA encapsulated by liposomes is absorbed by dendritic cells (DCs). Within DCs, mRNA undergoes the steps of transcription and translation to generate antigens, which are subsequently presented to T cells through MHC I or MHC II. Moreover, through the synergistic action of cytokines including interleukin-1 (IL-1), interleukin-2 (IL-2), and interleukin-12 (IL-12), the cellular immune pathway is activated. In particular, antigens secreted by APCs can also activate B lymphocytes. Under the action of activated CD4<sup>+</sup> T cells, B lymphocytes will secrete corresponding neutralizing antibodies, further enhancing the immune effect on tumors. Immune activation progress: ① vaccine internalization, ② and ③ encoding protein, ④ protein delivery, ⑤, ⑥ and ⑦ activating immunity.</p>
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17 pages, 4833 KiB  
Article
Fabrication and Properties of Hydrogel Dressings Based on Genipin Crosslinked Chondroitin Sulfate and Chitosan
by Ling Wang, Xiaoyue Ding, Xiaorui He, Ning Tian, Peng Ding, Wei Guo, Oseweuba Valentine Okoro, Yanfang Sun, Guohua Jiang, Zhenzhong Liu, Armin Shavandi and Lei Nie
Polymers 2024, 16(20), 2876; https://doi.org/10.3390/polym16202876 - 11 Oct 2024
Viewed by 475
Abstract
Multifunctional hydrogel dressings remain highly sought after for the promotion of skin wound regeneration. In the present study, multifunctional CHS-DA/HACC (CH) hydrogels with an interpenetrated network were constructed using hydroxypropyl trimethyl ammonium chloride modified chitosan (HACC) and dopamine-modified chondroitin sulfate (CHS-DA), using genipin [...] Read more.
Multifunctional hydrogel dressings remain highly sought after for the promotion of skin wound regeneration. In the present study, multifunctional CHS-DA/HACC (CH) hydrogels with an interpenetrated network were constructed using hydroxypropyl trimethyl ammonium chloride modified chitosan (HACC) and dopamine-modified chondroitin sulfate (CHS-DA), using genipin as crosslinker. The synthesis of HACC and CHS-DA was effectively confirmed using Fourier transform infrared (FT-IR) analysis and 1H nuclear magnetic resonance (1H NMR) spectroscopy. The prepared CH hydrogels exhibited a network of interconnected pores within the microstructure. Furthermore, rheological testing demonstrated that CH hydrogels exhibited strong mechanical properties, stability, and injectability. Further characterization investigations showed that the CH hydrogels showed favorable self-healing and self-adhesion properties. It was also shown that increasing HACC concentration ratio was positively correlated with the antibacterial activity of CH hydrogels, as evidenced by their resistance to Escherichia coli and Staphylococcus aureus. Additionally, Cell Counting Kit-8 (CCK-8) tests, fluorescent images, and a cell scratch assay demonstrated that CH hydrogels had good biocompatibility and cell migration ability. The multifunctional interpenetrated network hydrogels were shown to have good antibacterial properties, antioxidant properties, stable storage modulus and loss modulus, injectable properties, self-healing properties, and biocompatibility, highlighting their potential as wound dressings in wound healing applications. Full article
(This article belongs to the Special Issue Bioactive and Biomedical Hydrogel Dressings for Wound Healing)
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<p>(<b>a</b>) Chondroitin sulfate (CHS), DA, and CHS-DA FTIR spectra. (<b>b</b>) <sup>1</sup>H NMR spectra comparing CHS with the synthesized CHS-DA. (<b>c</b>) Chitosan and HACC FT-IR spectra. (<b>d</b>) The spectra (<sup>1</sup>H NMR) of chitosan and the produced HACC.</p>
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<p>Characterization of the synthesized CH hydrogels: (<b>a</b>) FT-IR spectra of CH hydrogels. (<b>b</b>) Stress–strain profiles of CH hydrogels. (<b>c</b>) SEM images of CH hydrogels at various magnifications. (<b>d</b>) Pore size distribution of CH hydrogels was calculated from SEM images using ImageJ software (1.54g version). (<b>e</b>) Equilibrium swelling ratio of CH hydrogels in PBS. * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Rheological analysis of the CH hydrogels: the storage modulus (G′) and loss modulus (G″) were measured as functions of frequency (<b>a</b>–<b>c</b>) and time (<b>d</b>–<b>f</b>). In addition, the frequency was kept at 1 Hz during G′ and G″ testing over time. Panels (<b>g</b>–<b>i</b>) show the variations in viscosity of the CH hydrogels with shear rate, and viscosity could reflect a fluid’s resistance to a change in shape or movement. The inset images illustrate that the CH hydrogels can be injected using a syringe.</p>
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<p>(<b>a</b>) A three-step cyclic strain test was conducted using a rheometer to assess the self-healing capabilities of the CH hydrogels. (<b>b</b>) The self-healing properties of the CH hydrogels were further examined through macroscopic observation, where the hydrogels were cut into two pieces and rejoined, and their contact morphology was documented. (<b>c</b>) Images showing the adhesion of CH hydrogels to various substrates, including glass, human skin, rubber, plastic, wood, and metal.</p>
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<p>(<b>a</b>) Images showing the inhibition zones created by CH hydrogels against <span class="html-italic">E. coli</span> and <span class="html-italic">S. aureus</span> after 8 h, demonstrating their antibacterial properties. Note that the left and right images represent repeated experiments. (<b>b</b>,<b>c</b>) The radius of the inhibition zones for CH hydrogels against (<b>b</b>) <span class="html-italic">S. aureus</span> and (<b>c</b>) <span class="html-italic">E. coli</span>, calculated based on the observed zones. (<b>d</b>) DPPH radical scavenging rate of CH hydrogels within 30 min, highlighting their antioxidant activity. * <span class="html-italic">p</span> &lt; 0.05 and *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Biocompatibility and cell migration of CH hydrogels. (<b>a</b>) Fluorescent microscopy images of NIH-3T3 cells cultured with CH hydrogel extracts for 1, 3, and 5 days, respectively. Cells were stained with Calcein-AM/PI; scale bars: 200 μm. (<b>b</b>) Hydrogel cytocompatibilities were assessed with NIH-3T3 cells at various time points using the CCK-8 assay. (<b>c</b>) Scratch assay results for NIH-3T3 cells after 8 h of incubation, with blue dotted lines representing the width of the scratch gap. Scale bar: 100 μm. (<b>d</b>) The migration rate of NIH-3T3 cells based on scratch assay results. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>The schematic representation of CH hydrogel preparation: (<b>a</b>) Synthesis of dopamine-modified chondroitin sulfate (CHS-DA), (<b>b</b>) Production of hydroxypropyl trimethyl ammonium chloride-modified chitosan (HACC), and (<b>c</b>) Fabrication of CH hydrogel with an interpenetrated network structure via genipin crosslinking polymers of HACC and CHS-DA.</p>
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21 pages, 6513 KiB  
Article
A Monitoring Device and Grade Prediction System for Grain Mildew
by Lei Xu, Yane Li, Xiang Weng, Jiankai Shi, Hailin Feng, Xingquan Liu and Guoxin Zhou
Sensors 2024, 24(20), 6556; https://doi.org/10.3390/s24206556 - 11 Oct 2024
Viewed by 296
Abstract
Mildew infestation is a significant cause of loss during grain storage. The growth and metabolism of mildew leads to changes in gas composition and temperature within granaries. Recent advances in sensor technology and machine learning enable the prediction of grain mildew during storage. [...] Read more.
Mildew infestation is a significant cause of loss during grain storage. The growth and metabolism of mildew leads to changes in gas composition and temperature within granaries. Recent advances in sensor technology and machine learning enable the prediction of grain mildew during storage. Current research primarily focuses on predicting mildew occurrence or grading using simple machine learning methods, without in-depth exploration of the time series characteristics of mildew process data. A monitoring device was designed and developed to capture high-quality microenvironment parameters and image data during a simulated mildew process experiment. Using the “Yongyou 15” rice varieties from Zhejiang Province, five simulation experiments were conducted under varying temperature and humidity conditions between January and May 2023. Mildew grades were defined through manual analysis to construct a multimodal dataset for the rice mildew process. This study proposes a combined model (CNN–LSTM–A) that integrates convolutional neural networks (CNN), long short-term memory (LSTM) networks, and attention mechanisms to predict the mildew grade of stored rice. The proposed model was compared with LSTM, CNN–LSTM, and LSTM–Attention models. The results indicate that the proposed model outperforms the others, achieving a prediction accuracy of 98%. The model demonstrates superior accuracy and more stable performance. The generalization performance of the prediction model was evaluated using four experimental datasets with varying storage temperature and humidity conditions. The results show that the model achieves optimal prediction stability when the training set contains similar storage temperatures, with prediction accuracy exceeding 99.8%. This indicates that the model can effectively predict the mildew grades in rice under varying environmental conditions, demonstrating significant potential for grain mildew prediction and early warning systems. Full article
(This article belongs to the Section Sensor Networks)
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<p>Flowchart of rice mildew grade prediction.</p>
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<p>System block diagram of grain mildew process monitoring device.</p>
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<p>Schematic diagram of data acquisition in the experimental chamber: (<b>a</b>) funnel opening for water filling, (<b>b</b>) microenvironment sensor deployment in the experimental box, (<b>c</b>) top camera.</p>
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<p>Eight consecutive days image dataset of rice mildew process. Dn denotes the nth day.</p>
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<p>Experimental process dataset.</p>
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<p>Correlation coefficient between grain parameters and mildew state.</p>
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<p>Labeling of rice mildew grade.</p>
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<p>Framework of CNN–LSTM–A prediction algorithm.</p>
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<p>Comparison of model losses with different: (<b>a</b>) epochs, (<b>b</b>) learning rates, (<b>c</b>) batch sizes and (<b>d</b>) hidden_size.</p>
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<p>Fitness curves of different models.</p>
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<p>Prediction results of different models.</p>
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<p>Comparison of model prediction confusion matrices under different datasets: (<b>a</b>) dataset 2, (<b>b</b>) dataset 3, (<b>c</b>) dataset 4, and (<b>d</b>) dataset 5.</p>
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<p>Comparison of model prediction results under different datasets: (<b>a</b>) dataset 2, (<b>b</b>) dataset 3, (<b>c</b>) dataset 4, and (<b>d</b>) dataset 5.</p>
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<p>Comparison of model prediction confusion matrices under different test sets: (<b>a</b>) test set 2, (<b>b</b>) test set 3, (<b>c</b>) test set 4, and (<b>d</b>) test set 5.</p>
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<p>Comparison of model prediction results under different test sets: (<b>a</b>) test set 2, (<b>b</b>) test set 3, (<b>c</b>) test set 4, and (<b>d</b>) test set 5.</p>
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10 pages, 6207 KiB  
Article
Remote Sulfonylation of Anilines with Sodium Sulfifinates Using Biomass-Derived Copper Catalyst
by Xiaoping Yan, Jinguo Wang, Chao Chen, Kai Zheng, Pengfei Zhang and Chao Shen
Molecules 2024, 29(20), 4815; https://doi.org/10.3390/molecules29204815 - 11 Oct 2024
Viewed by 290
Abstract
A biomass-based catalyst, CuxOy@CS-400, was employed as an excellent recyclable heterogeneous catalyst to realize the sulfonylation reaction of aniline derivatives with sodium sulfinates. Various substrates were compatible, giving the desired products moderate to good yields at room temperature. In [...] Read more.
A biomass-based catalyst, CuxOy@CS-400, was employed as an excellent recyclable heterogeneous catalyst to realize the sulfonylation reaction of aniline derivatives with sodium sulfinates. Various substrates were compatible, giving the desired products moderate to good yields at room temperature. In addition, this heterogeneous copper catalyst was also easy to recover and was recyclable up to five times without considerably deteriorating in catalytic efficiency. Importantly, these sulfonylation products were readily converted to the corresponding 4-sulfonyl anilines via a hydrolysis step. The method offers a unique strategy for synthesizing arylsulfones and has the potential to create new possibilities for developing heterogeneous copper-catalyzed C-H functionalizations. Full article
(This article belongs to the Special Issue Two-Dimensional Materials: From Synthesis to Applications)
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<p>Examples of aryl sulfone-based bioactive molecules.</p>
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<p>SEM images of Cu<sub>x</sub>O<sub>y</sub>@CS-400 (<b>a</b>,<b>b</b>). TEM images of Cu<sub>x</sub>O<sub>y</sub>@CS-400 (<b>c</b>,<b>d</b>). Elemental mapping of Cu<sub>x</sub>O<sub>y</sub>@CS-400 (<b>e</b>–<b>j</b>).</p>
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<p>Catalyst reutilization studies.</p>
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<p>Direct sulfonylation of 1-naphthylamides or anilines at the C4 site.</p>
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<p>Synthesis of Cu<sub>x</sub>O<sub>y</sub>@CS-T catalyst.</p>
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<p>Substrate scope of anilines and sodium sulfonates. Reaction conditions: anilines (0.2 mmol), sodium sulfonates (2.0 equiv.), Cu<sub>x</sub>O<sub>y</sub>@CS-400 (20 mg.), Ag<sub>2</sub>CO<sub>3</sub> (20 mol%), K<sub>2</sub>S<sub>2</sub>O<sub>8</sub> (2.0 equiv.), acetone/H<sub>2</sub>O (1:1) (3 mL), stirred at rt, under air, 12 h. Isolated yields.</p>
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<p>Synthetic applications and control experiment. (<b>a</b>) Gram-scale synthesis; (<b>b</b>) Radical trapping experiment.</p>
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<p>Proposed mechanism.</p>
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9 pages, 2316 KiB  
Article
Highly Efficient Organic/Silicon Hybrid Solar Cells with a MoO3 Capping Layer
by Jiahui Chen, Zhangbo Lu, Xiaoting Wang, Yuner Luo, Yun Ma, Gang Lou, Dan Chi and Shihua Huang
Nanomaterials 2024, 14(20), 1630; https://doi.org/10.3390/nano14201630 - 11 Oct 2024
Viewed by 515
Abstract
Organic/Si hybrid solar cells have attracted considerable attention for their uncomplicated fabrication process and superior device efficiency, making them a promising candidate for sustainable energy applications. However, the efficient collection and separation of charge carriers at the organic/Si heterojunction interface are primarily hindered [...] Read more.
Organic/Si hybrid solar cells have attracted considerable attention for their uncomplicated fabrication process and superior device efficiency, making them a promising candidate for sustainable energy applications. However, the efficient collection and separation of charge carriers at the organic/Si heterojunction interface are primarily hindered by the inadequate work function of poly (3,4-ethylenedioxythiophene): poly (styrenesulfonate) (PEDOT:PSS). Here, the application of a high-work-function MoO3 film onto the n-Si/PEDOT:PSS surface leads to a notable enhancement in the device’s built-in potential. This enhancement results in the creation of an inversion layer near the n-Si surface and facilitates charge separation at the interface. Simultaneously, it inhibits charge recombination at the heterojunction interface. As a result, the champion PEDOT:PSS/Si solar cell, which incorporates a MoO3 interface layer, demonstrates an efficiency of 16.0% and achieves a high fill factor of 80.8%. These findings provide a straightforward and promising strategy for promoting the collection and transmission of charge carriers at the interface of photovoltaic devices. Full article
(This article belongs to the Special Issue Low-Dimensional Perovskite Materials and Devices)
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<p>(<b>a</b>) Schematic illustration of the thermal evaporation deposition process of the MoO<sub>3</sub>. (<b>b</b>) The Mo 3d core level of evaporated MoO<sub>3</sub> films.</p>
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<p>(<b>a</b>) Device configuration of PEDOT:PSS/Si solar cells with a MoO<sub>3</sub> capping layer. (<b>b</b>) J–V curves of PEDOT:PSS/Si solar cells without and with the MoO<sub>3</sub> layer under simulated AM 1.5 illumination at 100 mW cm<sup>−2</sup>. (<b>c</b>) Corresponding dark J−V and (<b>d</b>) EQE curves of PEDOT:PSS/Si solar cells. The historical (<b>e</b>) V<sub>OC</sub> and (<b>f</b>) FF of high-performance planar Si/PEDOT:PSS solar cells with a modified front interface layer are juxtaposed for comparison.</p>
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<p>(<b>a</b>) UPS measured work function of the as-prepared PEDOT:PSS, PEDOT:PSS/MoO<sub>3</sub>, and MoO<sub>3</sub> films. (<b>b</b>) Capacitance−voltage measurements of the Si/PEDOT:PSS heterojunction solar cells without and with the thin layer of MoO<sub>3</sub>. (<b>c</b>) Energy band diagram of the PEDOT:PSS/Si hybrid solar cells.</p>
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<p>Surface potential image of MoO<sub>3</sub> film partially covering the surface of PEDOT:PSS (<b>a</b>) without and (<b>c</b>) with solar irradiation. Both of the scale bars are 10 µm. Cross-sectional line profile of the surface potential image (<b>b</b>) without and (<b>d</b>) with solar irradiation.</p>
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<p>Characterization of effective carrier lifetime. (<b>a</b>) The schematic of the lifetime measurements using a Sinton Instruments WCT-120 photo-conductance tool and lifetime test structure. (<b>b</b>) The measured effective carrier lifetime as a function of excess carrier density for n-type crystalline silicon wafers symmetrically passivated without and with MoO<sub>3</sub> films.</p>
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18 pages, 1566 KiB  
Review
The Role of Soluble CD163 (sCD163) in Human Physiology and Pathophysiology
by Andriana Plevriti, Margarita Lamprou, Eleni Mourkogianni, Nikolaos Skoulas, Maria Giannakopoulou, Md Sanaullah Sajib, Zhiyong Wang, George Mattheolabakis, Antonios Chatzigeorgiou, Antonia Marazioti and Constantinos M. Mikelis
Cells 2024, 13(20), 1679; https://doi.org/10.3390/cells13201679 - 11 Oct 2024
Viewed by 591
Abstract
Soluble CD163 (sCD163) is a circulating inflammatory mediator, indicative of acute and chronic, systemic and non-systemic inflammatory conditions. It is the cleavage outcome, consisting of almost the entire extracellular domain, of the CD163, a receptor expressed in monocytic lineages. Its expression is proportional [...] Read more.
Soluble CD163 (sCD163) is a circulating inflammatory mediator, indicative of acute and chronic, systemic and non-systemic inflammatory conditions. It is the cleavage outcome, consisting of almost the entire extracellular domain, of the CD163, a receptor expressed in monocytic lineages. Its expression is proportional to the abundance of CD163+ macrophages. Various mechanisms trigger the shedding of the CD163 receptor or the accumulation of CD163-expressing macrophages, inducing the sCD163 concentration in the circulation and bodily fluids. The activities of sCD163 range from hemoglobin (Hb) scavenging, macrophage marker, decoy receptor for cytokines, participation in immune defense mechanisms, and paracrine effects in various tissues, including the endothelium. It is an established marker of macrophage activation and thus participates in many diseases, including chronic inflammatory conditions, such as atherosclerosis, asthma, and rheumatoid arthritis; acute inflammatory conditions, such as sepsis, hepatitis, and malaria; insulin resistance; diabetes; and tumors. The sCD163 levels have been correlated with the severity, stage of the disease, and clinical outcome for many of these conditions. This review article summarizes the expression and role of sCD163 and its precursor protein, CD163, outlines the sCD163 generation mechanisms, the biological activities, and the known underlying molecular mechanisms, with an emphasis on its impact on the endothelium and its contribution in the pathophysiology of human diseases. Full article
(This article belongs to the Special Issue Immune Cell Effect on the Endothelium)
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<p>Schematic representation of the three CD163 transmembrane isoforms. They differ in the length of the intracellular domains, with the 49-amino-acid isoform having dominant expression.</p>
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<p>(<b>A</b>) Uniform manifold approximation and projection (UMAP) representation of all human cell types based on CD163 expression (data from 483,152 cells from the CZ database). (<b>B</b>) Clustering and analysis of the cell types with the highest CD163 expression.</p>
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<p>Schematic representation of the hemoglobin clearance mechanism by CD163 in inflammatory conditions. The Hb released by the ruptured red cells is bound to the 3rd SRCR domain of the CD163 extracellular domain, either by itself or as a Hp–Hb complex, which eventually leads to CD163 internalization.</p>
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<p>Schematic representation of sCD163 generation from the proteolytic cleavage of CD163. The list of the proteases known to cleave CD163 is shown on the left and the cleavage requires ATP consumption (black arrow).</p>
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10 pages, 722 KiB  
Article
Presence of N, N′-Substituted p-Phenylenediamine-Derived Quinones in Human Urine
by Juxiu Huang, Hangbiao Jin, Yingying Zhu, Ruyue Guo, Lisha Zhou and Xiaoyu Wu
Toxics 2024, 12(10), 733; https://doi.org/10.3390/toxics12100733 - 11 Oct 2024
Viewed by 409
Abstract
Human exposure to various N,N′-substituted p-phenylenediamine-derived quinones (PPDQs) has been of increasing concern. Recent studies have examined N-phenyl-N′-(1,3-dimethylbutyl)-p-phenylenediamine-derived quinone (6PPDQ) in human urine to evaluate human exposure. However, other PPDQs in human urine have not been thoroughly investigated. This study analyzed six PPDQs [...] Read more.
Human exposure to various N,N′-substituted p-phenylenediamine-derived quinones (PPDQs) has been of increasing concern. Recent studies have examined N-phenyl-N′-(1,3-dimethylbutyl)-p-phenylenediamine-derived quinone (6PPDQ) in human urine to evaluate human exposure. However, other PPDQs in human urine have not been thoroughly investigated. This study analyzed six PPDQs in urine collected from 149 healthy individuals in Taizhou, China. All target PPDQs were detected, with 6PPDQ (mean 2.4 ng/mL, <limit of detection (LOD)–19 ng/mL) and 2-(cyclohexylamino)-5-(phenylamino)cyclohexa-2,5-diene-1,4-dione (CPPDQ; 2.1 ng/mL, <LOD–24 ng/mL) being the most prevalent. Human urinary concentrations of 2,5-bis((5-methylhexan-2-yl)amino)cyclohexa-2,5-diene-1,4-dione (77PDQ; mean 1.5 vs. 0.87 ng/mL; p = 0.013) and 2,5-bis(o-tolylamino)cyclohexa-2,5-diene-1,4-dione (mean 1.1 vs. 0.62 ng/mL; p = 0.027) were significantly higher in females compared to males. For CPPDQ (p < 0.01) and 6PPDQ (p < 0.01), a decrease was observed in urinary concentrations as participants aged. The daily excretion (DE) of PPDQs through urine was estimated for Chinese adults. The highest average DE was recorded for 6PPDQ at 81 ng/kg-bw/day, with a range from <0.5 to 475 ng/kg-bw/day. Following this, CPPDQ had a mean DE of 68 ng/kg-bw/day (range <0.5–516 ng/kg-bw/day), and 77PDQ had a mean DE of 30 ng/kg-bw/day (<0.5–481 ng/kg-bw/day). This study is the first to explore the presence of various PPDQs in human urinary samples, which is essential for assessing the potential health risks associated with these substances. Full article
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<p>Composition profiles of PPDQs in human urine (<span class="html-italic">n</span> = 149) from Taizhou, China.</p>
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<p>Concentrations (mean ± SD) of PPDQs in urine samples from male and female participants. The red asterisk indicates the significant (<span class="html-italic">p</span> &lt; 0.05) difference in the human urinary concentration between male and female participants.</p>
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<p>Concentrations (mean ± SD) of PPDQs in human urine from participants among different age groups.</p>
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15 pages, 1713 KiB  
Article
Genome-Wide and Exome-Wide Association Study Identifies Genetic Underpinning of Comorbidity between Myocardial Infarction and Severe Mental Disorders
by Bixuan Jiang, Xiangyi Li, Mo Li, Wei Zhou, Mingzhe Zhao, Hao Wu, Na Zhang, Lu Shen, Chunling Wan, Lin He, Cong Huai and Shengying Qin
Biomedicines 2024, 12(10), 2298; https://doi.org/10.3390/biomedicines12102298 - 10 Oct 2024
Viewed by 494
Abstract
Background: Myocardial Infarction (MI) and severe mental disorders (SMDs) are two types of highly prevalent and complex disorders and seem to have a relatively high possibility of mortality. However, the contributions of common and rare genetic variants to their comorbidity arestill unclear. Methods: [...] Read more.
Background: Myocardial Infarction (MI) and severe mental disorders (SMDs) are two types of highly prevalent and complex disorders and seem to have a relatively high possibility of mortality. However, the contributions of common and rare genetic variants to their comorbidity arestill unclear. Methods: We conducted a combined genome-wide association study (GWAS) and exome-wide association study (EWAS) approach. Results: Using gene-based and gene-set association analyses based on the results of GWAS, we found the common genetic underpinnings of nine genes (GIGYF2, KCNJ13, PCCB, STAG1, HLA-C, HLA-B, FURIN, FES, and SMG6) and nine pathways significantly shared between MI and SMDs. Through Mendelian randomization analysis, we found that twenty-seven genes were potential causal genes for SMDs and MI. Based on the exome sequencing data of MI and SMDs patients from the UK Biobank, we found that MUC2 was exome-wide significant in the two diseases. The gene-set analyses of the exome-wide association study indicated that pathways related to insulin processing androgen catabolic process and angiotensin receptor binding may be involved in the comorbidity between SMDs and MI. We also found that six candidate genes were reported to interact with known therapeutic drugs based on the drug–gene interaction information in DGIdb. Conclusions: Altogether, this study revealed the overlap of common and rare genetic underpinning between SMDs and MI and may provide useful insights for their mechanism study and therapeutic investigations. Full article
(This article belongs to the Section Molecular Genetics and Genetic Diseases)
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<p>Shared gene sets among Myocardial Infarction and severe mental disorders in GWAS analyses. Gene sets enriched in MAGMA analyses of the BD (<b>A</b>), SCZ (<b>B</b>), and MI (<b>C</b>) significant common architecture are shown in the plots. The dots colored red represent significant common gene sets based on a Benjamini–Hochberg correction for multiple testing at FDR &lt; 0.25 and nominal <span class="html-italic">p</span> value &lt; 0.05. The data are available in <a href="#app1-biomedicines-12-02298" class="html-app">Supplementary Tables S1–S3</a>. The specific term names of gene sets from GO or KEGG are listed below. GO:0048878, chemical homeostasis; GO:0001505, regulation of neurotransmitter levels; GO:0006836, neurotransmitter transport; GO:0034364, high-density lipoprotein particle; GO:0042627, chylomicron; GO:0034385, triglyceride-rich plasma lipoprotein particle; GO:0140059, dendrite arborization; GO:0051940, regulation of catecholamine uptake involved in synaptic transmission; PATHWAY: hsa04720, long-term potentiation. BD: Bipolar Disorder; SCZ: Schizophrenia; MI: Myocardial Infarction.</p>
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<p>QQ plots for gene-level association tests in WES analyses. Genes enriched in RVtests analyses of the BD (<b>A</b>), SCZ (<b>B</b>), and MI (<b>C</b>) significant rare architecture are shown in the plots. The dots colored red represent statistically significant genes based on a Benjamini–Hochberg correction for multiple testing at FDR &lt; 0.25 and <span class="html-italic">p</span> value &lt; 0.05. BD: Bipolar Disorder; SCZ: Schizophrenia; MI: Myocardial Infarction. The data are available in <a href="#app1-biomedicines-12-02298" class="html-app">Supplementary Tables S10–S12</a>.</p>
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<p>Gene–drug interactions for BD, SCZ, and MI. The diagram reveals the correlation between the disorders, genes, and drugs. BD: Bipolar Disorder; SCZ: Schizophrenia; MI: Myocardial Infarction. The data are available in <a href="#app1-biomedicines-12-02298" class="html-app">Supplementary Table S16</a>.</p>
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19 pages, 4642 KiB  
Article
SARS-CoV-2 Nucleocapsid Protein Antagonizes GADD34-Mediated Innate Immune Pathway through Atypical Foci
by Jie Liu, Guanwen Guan, Chunxiu Wu, Bingbing Wang, Kaifei Chu, Xu Zhang, Su He, Naru Zhang, Geng Yang, Zhigang Jin and Tiejun Zhao
Molecules 2024, 29(20), 4792; https://doi.org/10.3390/molecules29204792 - 10 Oct 2024
Viewed by 591
Abstract
The integrated stress response, especially stress granules (SGs), contributes to host immunity. Typical G3BP1+ stress granules (tSGs) are usually formed after virus infection to restrain viral replication and stimulate innate immunity. Recently, several SG-like foci or atypical SGs (aSGs) with proviral function [...] Read more.
The integrated stress response, especially stress granules (SGs), contributes to host immunity. Typical G3BP1+ stress granules (tSGs) are usually formed after virus infection to restrain viral replication and stimulate innate immunity. Recently, several SG-like foci or atypical SGs (aSGs) with proviral function have been found during viral infection. We have shown that the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid (N) protein induces atypical N+/G3BP1+ foci (N+foci), leading to the inhibition of host immunity and facilitation of viral infection. However, the precise mechanism has not been well clarified yet. In this study, we showed that the SARS-CoV-2 N (SARS2-N) protein inhibits dsRNA-induced growth arrest and DNA damage-inducible 34 (GADD34) expression. Mechanistically, the SARS2-N protein promotes the interaction between GADD34 mRNA and G3BP1, sequestering GADD34 mRNA into the N+foci. Importantly, we found that GADD34 participates in IRF3 nuclear translocation through its KVRF motif and promotes the transcription of downstream interferon genes. The suppression of GADD34 expression by the SARS2-N protein impairs the nuclear localization of IRF3 and compromises the host’s innate immune response, which facilitates viral replication. Taking these findings together, our study revealed a novel mechanism by which the SARS2-N protein antagonized the GADD34-mediated innate immune pathway via induction of N+foci. We think this is a critical strategy for viral pathogenesis and has potential therapeutic implications. Full article
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Figure 1
<p>SARS2-N protein inhibits dsRNA-induced GADD34 expression. (<b>A</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and treated with poly I:C for the indicated time. Cell lysates were subjected to Western blot to detect eIF2α, p-eIF2α (S51), GADD34, FLAG-SARS2-N, and GAPDH. (<b>B</b>) Graphical representation of the relative amount of GADD34 to GAPDH shown in panel (<b>A</b>). The bars indicate the mean ± SD. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>C</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and were untreated or treated with poly I:C for the indicated time. An RT-qPCR experiment was performed to detect the mRNA level of GADD34 and 18S RNA. (<b>D</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and poly I:C. After treatment with 10 μM of MG132, cell lysates were subjected to Western blot to detect GADD34, FLAG-SARS2-N, and GAPDH. (<b>E</b>) Graphical representation of the relative amount of GADD34 to GAPDH is shown in panel D. Bars indicate mean ± SD. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>F</b>) 16HBE cells were transfected with pGL3-SV40, GADD34 5′-UTR, GADD34 uORF1-AUA, or GADD34 uORF2-AUA reporter plasmids (SV40-Luc, GADD34 5′-UTR-Luc, GADD34 uORF1-AUA-Luc, and GADD34 uORF2-AUA-Luc) and expression vectors for SARS2-N (pCS2-Flag-SARS2-N) After treating the cells with poly I:C for 9 h, a dual-luciferase assay was performed. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01).</p>
Full article ">Figure 1 Cont.
<p>SARS2-N protein inhibits dsRNA-induced GADD34 expression. (<b>A</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and treated with poly I:C for the indicated time. Cell lysates were subjected to Western blot to detect eIF2α, p-eIF2α (S51), GADD34, FLAG-SARS2-N, and GAPDH. (<b>B</b>) Graphical representation of the relative amount of GADD34 to GAPDH shown in panel (<b>A</b>). The bars indicate the mean ± SD. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>C</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and were untreated or treated with poly I:C for the indicated time. An RT-qPCR experiment was performed to detect the mRNA level of GADD34 and 18S RNA. (<b>D</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and poly I:C. After treatment with 10 μM of MG132, cell lysates were subjected to Western blot to detect GADD34, FLAG-SARS2-N, and GAPDH. (<b>E</b>) Graphical representation of the relative amount of GADD34 to GAPDH is shown in panel D. Bars indicate mean ± SD. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>F</b>) 16HBE cells were transfected with pGL3-SV40, GADD34 5′-UTR, GADD34 uORF1-AUA, or GADD34 uORF2-AUA reporter plasmids (SV40-Luc, GADD34 5′-UTR-Luc, GADD34 uORF1-AUA-Luc, and GADD34 uORF2-AUA-Luc) and expression vectors for SARS2-N (pCS2-Flag-SARS2-N) After treating the cells with poly I:C for 9 h, a dual-luciferase assay was performed. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01).</p>
Full article ">Figure 2
<p>SARS2-N promotes the association of GADD34 mRNA with N<sup>+</sup>foci. (<b>A</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and/or poly I:C followed by RIP with an anti-G3BP1 antibody. The enrichment of <span class="html-italic">GADD34</span> mRNA by G3BP1 was assessed by RT-qPCR and shown as the relative amount of <span class="html-italic">GADD34</span> mRNAs in immunoprecipitates compared to that of 5% input. Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05). (<b>B</b>) Cell lysates in the RIP assay were subjected to Western blot to detect GADD34, FLAG-SARS2-N, and GAPDH. (<b>C</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and/or poly I:C together with pCS2-GFP-G3BP1-TurboID expression vector. Cells were harvested after the treatment of biotin for 15 min. mRNA associated with the biotinylated tSG protein or N<sup>+</sup>foci proteins were enriched with streptavidin beads followed by RT-qPCR for <span class="html-italic">GADD34</span> mRNA. Data are shown as the relative amount of <span class="html-italic">GADD34</span> mRNAs in streptavidin beads compared to that of 5% input. Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05). (<b>D</b>) Cell lysates in the TurboID assay were subjected to Western blot to detect GADD34, FLAG-SARS2-N, and GAPDH. (<b>E</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and/or poly I:C together with cy5-labeled <span class="html-italic">GADD34</span> mRNA followed by immunostaining for Flag (green) and G3BP1 (red). In poly I:C-transfected cells, G3BP1<sup>+</sup> condensates indicate tSGs. In SARS2-N and poly I:C-transfected cells, N<sup>+</sup>G3BP1<sup>+</sup> condensates indicate N<sup>+</sup>foci. Scale bars: 20 μm. (<b>F</b>) Statistical analysis of the percentage of cells with tSGs or N<sup>+</sup>foci and the percentage of tSGs/N<sup>+</sup>foci that include <span class="html-italic">GADD34</span> mRNA is shown in panel (<b>E</b>). Data are shown as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test.</p>
Full article ">Figure 2 Cont.
<p>SARS2-N promotes the association of GADD34 mRNA with N<sup>+</sup>foci. (<b>A</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and/or poly I:C followed by RIP with an anti-G3BP1 antibody. The enrichment of <span class="html-italic">GADD34</span> mRNA by G3BP1 was assessed by RT-qPCR and shown as the relative amount of <span class="html-italic">GADD34</span> mRNAs in immunoprecipitates compared to that of 5% input. Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05). (<b>B</b>) Cell lysates in the RIP assay were subjected to Western blot to detect GADD34, FLAG-SARS2-N, and GAPDH. (<b>C</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and/or poly I:C together with pCS2-GFP-G3BP1-TurboID expression vector. Cells were harvested after the treatment of biotin for 15 min. mRNA associated with the biotinylated tSG protein or N<sup>+</sup>foci proteins were enriched with streptavidin beads followed by RT-qPCR for <span class="html-italic">GADD34</span> mRNA. Data are shown as the relative amount of <span class="html-italic">GADD34</span> mRNAs in streptavidin beads compared to that of 5% input. Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05). (<b>D</b>) Cell lysates in the TurboID assay were subjected to Western blot to detect GADD34, FLAG-SARS2-N, and GAPDH. (<b>E</b>) 16HBE cells were transfected with pCS2-Flag-SARS2-N and/or poly I:C together with cy5-labeled <span class="html-italic">GADD34</span> mRNA followed by immunostaining for Flag (green) and G3BP1 (red). In poly I:C-transfected cells, G3BP1<sup>+</sup> condensates indicate tSGs. In SARS2-N and poly I:C-transfected cells, N<sup>+</sup>G3BP1<sup>+</sup> condensates indicate N<sup>+</sup>foci. Scale bars: 20 μm. (<b>F</b>) Statistical analysis of the percentage of cells with tSGs or N<sup>+</sup>foci and the percentage of tSGs/N<sup>+</sup>foci that include <span class="html-italic">GADD34</span> mRNA is shown in panel (<b>E</b>). Data are shown as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test.</p>
Full article ">Figure 3
<p>SARS2-N suppresses GADD34-mediated innate immunity response. (<b>A</b>) HEK293T cells were untreated or pretreated with 10 or 40 μM of Guanabenz for 8 h, then transfected with IFN-β reporter plasmids (IFN-β-Luc) and treated with poly I:C for 9 h followed by luciferase assays. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>B</b>) HEK293T cells were untreated, treated with poly I:C, or treated with poly I:C and 40 μM of Guanabenz for 9 h, followed by RT-qPCR for IFN-β, IL-6, and ACTB. Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01). (<b>C</b>) HEK293T cells were transfected with IFN-β reporter plasmids (IFN-β-Luc) together with indicated plasmids (RIG-I, pCS2-Flag-SARS2-N, and Myc-GADD34), followed by luciferase assays. Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01). (<b>D</b>) HEK293T cells were transfected with indicated plasmids (RIG-I, pCS2-Flag-SARS2-N, and Myc-GADD34) followed by RT-qPCR for IFN-β, IFIT1, IFIT2, and ACTB. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>E</b>) HEK293T cells were transfected with Flag-IRF3 together with pCS2-YFP-SARS2-N and pCS2-Myc-GADD34 expression vectors and treated with poly I:C for 9 h, followed by immunostaining for Flag (red) and Myc (green). Scale bars: 20 μm. (<b>F</b>) Statistical analysis of the percentage of cells with nuclear IRF3 is shown in panel (<b>E</b>). Data are shown as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01).</p>
Full article ">Figure 4
<p>GADD34 barely affects SARS2-N protein-mediated induction of N<sup>+</sup>foci. (<b>A</b>) 16HBE cells were transfected with pCS2-YFP-SARS2-N and pCS2-Myc-GADD34 expression vectors and treated with poly I:C for 9 h, followed by immunostaining for G3BP1 (green) and Myc (red). Scale bars: 20 μm. (<b>B</b>) Statistical analysis of the percentage of cells with tSGs or N<sup>+</sup>foci is shown in panel A. Data are shown as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test. (<b>C</b>) 16HBE cells transfected with pCS2-Myc-G3BP1, pCS2-Flag-SARS2-N, and pCS2-Myc-GADD34 were subjected to immunoprecipitation with an anti-Flag antibody. The presence of G3BP1 in the immunoprecipitates was assessed by Western blot against anti-G3BP1 antibodies. (<b>D</b>) Quantification of immunoblot intensities is shown in (<b>C</b>). The intensity is the ratio of the G3BP1 band to the GAPDH band. Bars indicate mean ± SD (<span class="html-italic">n</span> = 3). Statistical analysis was performed with Student’s <span class="html-italic">t</span>-test (ns, no significance).</p>
Full article ">Figure 5
<p>GADD34 rescues innate immunity suppressed by SARS2-N protein through KVRF motif. (<b>A</b>) HEK293T cells transfected with Flag-PP1c, Myc-GADD34 WT, or Myc-GADD34 V556A/F558 were subjected to immunoprecipitation with an anti-Myc antibody. The presence of PP1c in the immunoprecipitates was assessed by Western blot against anti-Flag antibodies. (<b>B</b>) HEK293T cells were transfected with Myc-GADD34 WT or Myc-GADD34 V556A/F558 and treated with poly I:C for the indicated time. Cell lysates were subjected to Western blot to detect eIF2α, p-eIF2α (S51), Myc-GADD34, and GAPDH. (<b>C</b>) HEK293T cells were transfected with IFN-β reporter plasmids (IFN-β-Luc) together with pCS2-Flag-SARS2-N and pCS2-Myc-GADD34 or its mutant expression vectors. After treating the cells with poly I:C for 9 h, a dual-luciferase assay was performed. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>D</b>) HEK293T cells were transfected with pCS2-Flag-SARS2-N and pCS2-Myc-GADD34 or its mutant expression vectors, and treated with poly I:C for 9 h, followed by RT-qPCR for <span class="html-italic">IFN-β</span>, <span class="html-italic">IFIT1</span>, <span class="html-italic">IFIT2,</span> and <span class="html-italic">18S RNA</span>. Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01). (<b>E</b>) HEK293T cells were transfected with Flag-IRF3 together with pCS2-YFP-SARS2-N and pCS2-Myc-GADD34 or its mutant expression vectors, and treated with poly I:C for 9 h, followed by immunostaining for Flag (red) and Myc (green). Scale bars: 20 μm. (<b>F</b>) Statistical analysis of the percentage of cells with nuclear IRF3 is shown in panel (<b>E</b>). Data are shown as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01).</p>
Full article ">Figure 5 Cont.
<p>GADD34 rescues innate immunity suppressed by SARS2-N protein through KVRF motif. (<b>A</b>) HEK293T cells transfected with Flag-PP1c, Myc-GADD34 WT, or Myc-GADD34 V556A/F558 were subjected to immunoprecipitation with an anti-Myc antibody. The presence of PP1c in the immunoprecipitates was assessed by Western blot against anti-Flag antibodies. (<b>B</b>) HEK293T cells were transfected with Myc-GADD34 WT or Myc-GADD34 V556A/F558 and treated with poly I:C for the indicated time. Cell lysates were subjected to Western blot to detect eIF2α, p-eIF2α (S51), Myc-GADD34, and GAPDH. (<b>C</b>) HEK293T cells were transfected with IFN-β reporter plasmids (IFN-β-Luc) together with pCS2-Flag-SARS2-N and pCS2-Myc-GADD34 or its mutant expression vectors. After treating the cells with poly I:C for 9 h, a dual-luciferase assay was performed. Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>D</b>) HEK293T cells were transfected with pCS2-Flag-SARS2-N and pCS2-Myc-GADD34 or its mutant expression vectors, and treated with poly I:C for 9 h, followed by RT-qPCR for <span class="html-italic">IFN-β</span>, <span class="html-italic">IFIT1</span>, <span class="html-italic">IFIT2,</span> and <span class="html-italic">18S RNA</span>. Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01). (<b>E</b>) HEK293T cells were transfected with Flag-IRF3 together with pCS2-YFP-SARS2-N and pCS2-Myc-GADD34 or its mutant expression vectors, and treated with poly I:C for 9 h, followed by immunostaining for Flag (red) and Myc (green). Scale bars: 20 μm. (<b>F</b>) Statistical analysis of the percentage of cells with nuclear IRF3 is shown in panel (<b>E</b>). Data are shown as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01).</p>
Full article ">Figure 6
<p>GADD34 suppresses viral replication mainly by KVRF motif. (<b>A</b>) HEK293T cells were transfected with plasmids as indicated. Thirty-six hours later, cells were infected with VSV-GFP (MOI of 0.001). Twelve hours after virus infection, cells were collected for flow cytometry to detect the rate of GFP<sup>+</sup> cells and the supernatant was collected for the plaque assay. (<b>B</b>) Statistical analysis of the percentage of cells with VSV-GFP is shown in panel (<b>A</b>). The data are presented as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05 and **, <span class="html-italic">p</span> &lt; 0.01). (<b>C</b>) Vero-E6 cells were plated in 24-well plates for culture prior to the experiment. When the cells reached 100% confluency, the collected supernatant was diluted by a 10-fold gradient for infection. After 24 h, the number of plaques was counted and the virus titer was calculated. (<b>D</b>) Statistical analysis of VSV titers is shown in panel (<b>C</b>). The data are presented as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>E</b>) The proposed model that SARS2-N protein inhibits GADD34 expression and its physiological significance to the innate immune response and viral replication.</p>
Full article ">Figure 6 Cont.
<p>GADD34 suppresses viral replication mainly by KVRF motif. (<b>A</b>) HEK293T cells were transfected with plasmids as indicated. Thirty-six hours later, cells were infected with VSV-GFP (MOI of 0.001). Twelve hours after virus infection, cells were collected for flow cytometry to detect the rate of GFP<sup>+</sup> cells and the supernatant was collected for the plaque assay. (<b>B</b>) Statistical analysis of the percentage of cells with VSV-GFP is shown in panel (<b>A</b>). The data are presented as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test (*, <span class="html-italic">p</span> &lt; 0.05 and **, <span class="html-italic">p</span> &lt; 0.01). (<b>C</b>) Vero-E6 cells were plated in 24-well plates for culture prior to the experiment. When the cells reached 100% confluency, the collected supernatant was diluted by a 10-fold gradient for infection. After 24 h, the number of plaques was counted and the virus titer was calculated. (<b>D</b>) Statistical analysis of VSV titers is shown in panel (<b>C</b>). The data are presented as the mean ± SD (<span class="html-italic">n</span> = 3). Statistics: Student’s <span class="html-italic">t</span>-test (**, <span class="html-italic">p</span> &lt; 0.01). (<b>E</b>) The proposed model that SARS2-N protein inhibits GADD34 expression and its physiological significance to the innate immune response and viral replication.</p>
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