Coxsackievirus A6 Induces Cell Cycle Arrest in G0/G1 Phase for Viral Production

Recent epidemiological data indicate that outbreaks of hand, foot, and mouth disease (HFMD), which can be categorized according to its clinical symptoms as typical or atypical, have markedly increased worldwide. A primary causative agent for typical HFMD outbreaks, enterovirus 71 (EV71), has been shown to manipulate the cell cycle in S phase for own replication; however, it is not clear whether coxsackievirus (CVA6), the main agent for atypical HFMD, also regulates the host cell cycle. In this study, we demonstrate for the first time that CVA6 infection arrests the host cell cycle in G0/G1-phase. Furthermore, synchronization in G0/G1 phase, but not S phase or G2/M phase, promotes viral production. To investigate the mechanism of cell cycle arrest induced by CVA6 infection, we analyzed cell cycle progression after cell cycle synchronization at G0/G1 or G2/M. Our results demonstrate that CVA6 infection promotes G0/G1 phase entry from G2/M phase, and inhibits G0/G1 exit into S phase. In line with its role to arrest cells in G0/G1 phase, the expression of cyclinD1, CDK4, cyclinE1, CDK2, cyclinB1, CDK1, P53, P21, and P16 is regulated by CVA6. Finally, the non-structural proteins of CVA6, RNA-dependent RNA polymerase 3D and protease 3C , are demonstrated to be responsible for the G0/G1-phase arrest. These findings suggest that CVA6 infection arrested cell cycle in G0/G1-phase via non-structural proteins 3D and 3C, which may provide favorable environments for virus production

Crystallization and preliminary crystallographic analysis of thermophilic cellulase from Fervidobacterium nodosum Rt17-B1

The thermophilic cellulase FnCel5A from F. nodosum Rt17-B1 has been overexpressed in E. coli, purified and crystallized. Diffraction data were collected to 2.4 Å resolution for the native enzyme and to 1.7 Å resolution for the selenomethionyl derivative

Discriminative Semantic Feature Pyramid Network with Guided Anchoring for Logo Detection

Logo detection is a technology that identifies logos in images and returns their locations. With logo detection technology, brands can check how often their logos are displayed on social media platforms and elsewhere online and how they appear. It has received a lot of attention for its wide applications across different sectors, such as brand identity protection, product brand management, and logo duration monitoring. Particularly, logo detection technology can offer various benefits for companies to help brands measure their logo coverage, track their brand perception, secure their brand value, increase the effectiveness of their marketing campaigns and build brand awareness more effectively. However, compared with the general object detection, logo detection is more challenging due to the existence of both small logo objects and large aspect ratio logo objects. In this paper, we propose a novel approach, named Discriminative Semantic Feature Pyramid Network with Guided Anchoring (DSFP-GA), which can address these challenges via aggregating the semantic information and generating different aspect ratio anchor boxes. More specifically, our approach mainly consists of two components, namely Discriminative Semantic Feature Pyramid (DSFP) and Guided Anchoring (GA). The former is proposed to fuse semantic features into low-level feature maps to obtain discriminative representation of small logo objects, while the latter is further integrated into DSFP to generate large aspect ratio anchor boxes for detecting large aspect ratio logo objects. Extensive experimental results on four benchmarks demonstrate the effectiveness of the proposed DSFP-GA. Moreover, we further conduct visual analysis and ablation studies to illustrate the strength of the proposed DSFP-GA when detecting both small logo objects and large aspect logo objects

Long-Range Dependence Involutional Network for Logo Detection

Logo detection is one of the crucial branches in computer vision due to various real-world applications, such as automatic logo detection and recognition, intelligent transportation, and trademark infringement detection. Compared with traditional handcrafted-feature-based methods, deep learning-based convolutional neural networks (CNNs) can learn both low-level and high-level image features. Recent decades have witnessed the great feature representation capabilities of deep CNNs and their variants, which have been very good at discovering intricate structures in high-dimensional data and are thereby applicable to many domains including logo detection. However, logo detection remains challenging, as existing detection methods cannot solve well the problems of a multiscale and large aspect ratios. In this paper, we tackle these challenges by developing a novel long-range dependence involutional network (LDI-Net). Specifically, we designed a strategy that combines a new operator and a self-attention mechanism via rethinking the intrinsic principle of convolution called long-range dependence involution (LD involution) to alleviate the detection difficulties caused by large aspect ratios. We also introduce a multilevel representation neural architecture search (MRNAS) to detect multiscale logo objects by constructing a novel multipath topology. In addition, we implemented an adaptive RoI pooling module (ARM) to improve detection efficiency by addressing the problem of logo deformation. Comprehensive experiments on four benchmark logo datasets demonstrate the effectiveness and efficiency of the proposed approach

Impact of Chronic Diseases Follow-up on Health Behaviors and Blood Pressure/Glucose Control of Patients with Hypertension and Diabetes in the Context of Treatment-prevention Integration

Background Hypertension and diabetes are two major chronic diseases affecting population health, and need to be controlled through chronic diseases follow-up. However, there is currently insufficient understanding of the impact of different chronic diseases follow-up forms and contents on disease control and healthy behaviors of patients. Objective To explore the impact of the chronic diseases follow-up on health behaviors and blood pressure/glucose control of patients with hypertension and diabetes in the context of treatment-prevention integration. Methods Yiyang County in Henan Province, Xianfeng County in Hubei Province and Yangqu County in Shanxi Province were selected as study sites to collect data from the basic public health information system from 2017-01-01 to 2022-06-30. Patient survey was conducted in July 2022 to collect information on basic public health follow-up receiving, health behaviors and disease control of patients. Finally, 102 769 patients with hypertension and 26 586 patients with diabetes were obtained from the basic public health information system, the data of 1 172 patients with hypertension and 456 patients with diabetes were obtained through patient surveys. Multivariate Logistic regression was used to analyze the effects of the standard-reaching frequency of follow-up, follow-up forms and contents on health behaviors and disease control of patients. Results In 2021, the standard-reaching rates of follow-up frequency of patients with hypertension and diabetes were 90.83% (67 709/74 545) and 83.35% (13 390/16 065) , with frequency≥4 times/year as the standard. The follow-up forms included household follow-up〔25.74% (408/1 585) 〕, follow-up at institutional visits〔58.80% (932/1 585) 〕, telephone or network follow-up〔15.46% (245/1 585) 〕. The follow-up contents included blood pressure and blood glucose measurements〔91.15% (1 484/1 628) 〕, lifestyle guidance〔74.14% (1 207/1 628) 〕, disease inquiry〔70.02% (1 140/1 628) 〕, and drug use understanding〔69.29% (1 128/1 628) 〕. Multivariate Logistic regression analysis showed that patients with higher standard-reaching rates of follow-up frequency had higher rates of blood pressure control (OR=1.09, P<0.05) and glucose control (OR=1.31, P<0.05) , lower rates of smoking and drinking (OR=0.83, P<0.05) , and higher rates of regular exercise habits (OR=1.30, P<0.05) . The proportions of smoking and alcohol cessation (OR=2.38, P<0.05) and regular exercise habits (OR=1.62, P<0.05) were higher in the patients followed up at institutional visits than those followed up by telephone or network. The proportions of smoking and alcohol cessation (OR=2.33, P<0.05) and regular exercise habits (OR=2.54, P<0.05) of patients received household follow up were higher than those followed up by telephone or network. Patients who received lifestyle guidance, disease inquiry, and drug use understanding had higher rates of blood pressure control (OR=1.61, 1.34, and 1.62, respectively; P<0.05) , smoking and alcohol cessation (OR=3.59, 3.54, and 2.91, respectively; P<0.05) and regular exercise habits (OR=3.16, 2.15, 2.45, respectively; P<0.05) . Conclusion Receiving chronic diseases follow-up at least 4 times per year, with follow-up at institutional visits and household follow-up as the forms, provided with blood pressure and blood glucose measurements, lifestyle guidance, disease inquiry and drug use understanding as the contents in patients were positive correlated with blood pressure/glucose control, smoking and alcohol cessation, and regular exercise habits

Cloning, Expression and Characterization of a Novel Thermophilic Polygalacturonase from Caldicellulosiruptor bescii DSM 6725

We cloned the gene ACM61449 from anaerobic, thermophilic Caldicellulosiruptor bescii, and expressed it in Escherichia coli origami (DE3). After purification through thermal treatment and Ni-NTA agarose column extraction, we characterized the properties of the recombinant protein (CbPelA). The optimal temperature and pH of the protein were 72 °C and 5.2, respectively. CbPelA demonstrated high thermal-stability, with a half-life of 14 h at 70 °C. CbPelA also showed very high activity for polygalacturonic acid (PGA), and released monogalacturonic acid as its sole product. The Vmax and Km of CbPelA were 384.6 U·mg−1 and 0.31 mg·mL−1, respectively. CbPelA was also able to hydrolyze methylated pectin (48% and 10% relative activity on 20%–34% and 85% methylated pectin, respectively). The high thermo-activity and methylated pectin hydrolization activity of CbPelA suggest that it has potential applications in the food and textile industry

AZD1775 and anti-PD-1 antibody synergistically sensitize hepatoma to radiotherapy

Abstract. Background:. Radiation (IR)-induced DNA damage triggers cell cycle arrest and has a suppressive effect on the tumor microenvironment (TME). Wee1, a cell cycle regulator, can eliminate G2/M arrest by phosphorylating cyclin-dependent kinase 1 (CDK1). Meanwhile, programed death-1/programed death ligand-1 (PD-1/PDL-1) blockade is closely related to TME. This study aims to investigate the effects and mechanisms of Wee1 inhibitor AZD1775 and anti-PD-1 antibody (anti-PD-1 Ab) on radiosensitization of hepatoma. Methods:. The anti-tumor activity of AZD1775 and IR was determined by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay on human and mouse hepatoma cells HepG2, Hepa1-6, and H22. The anti-hepatoma mechanism of AZD1775 and IR revealed by flow cytometry and Western blot in vitro. A hepatoma subcutaneous xenograft mice model was constructed on Balb/c mice, which were divided into control group, IR group, AZD1775 group, IR + AZD1775 group, IR + anti-PD-1 Ab group, and the IR + AZD1775 + anti-PD-1 Ab group. Cytotoxic CD8+ T cells in TME were analyzed by flow cytometry. Results:. Combining IR with AZD1775 synergistically reduced the viability of hepatoma cells in vitro. AZD1775 exhibited antitumor effects by decreasing CDK1 phosphorylation to reverse the IR-induced G2/M arrest and increasing IR-induced DNA damage. AZD1775 treatment also reduced the proportion of PD-1+/CD8+ T cells in the spleen of hepatoma subcutaneous xenograft mice. Further studies revealed that AZD1775 and anti-PD-1 Ab could enhance the radiosensitivity of hepatoma by enhancing the levels of interferon γ (IFNγ)+ or Ki67+ CD8 T cells and decreasing the levels of CD8+ Tregs cells in the tumor and spleen of the hepatoma mice model, indicating that the improvement of TME was manifested by increasing the cytotoxic factor IFNγ expression, enhancing CD8+ T cells proliferation, and weakening CD8+ T cells depletion. Conclusions:. This work suggests that AZD1775 and anti-PD-1 Ab synergistically sensitize hepatoma to radiotherapy by enhancing IR-induced DNA damage and improving cytotoxic CD8+ T cells in TME

Deubiquitinase ubiquitin-specific protease 3 (USP3) inhibits HIV-1 replication via promoting APOBEC3G (A3G) expression in both enzyme activity-dependent and -independent manners

Abstract. Background:. Ubiquitination plays an essential role in many biological processes, including viral infection, and can be reversed by deubiquitinating enzymes (DUBs). Although some studies discovered that DUBs inhibit or enhance viral infection by various mechanisms, there is lack of information on the role of DUBs in virus regulation, which needs to be further investigated. Methods:. Immunoblotting, real-time polymerase chain reaction, in vivo/in vitro deubiquitination, protein immunoprecipitation, immunofluorescence, and co-localization biological techniques were employed to examine the effect of ubiquitin-specific protease 3 (USP3) on APOBEC3G (A3G) stability and human immunodeficiency virus (HIV) replication. To analyse the relationship between USP3 and HIV disease progression, we recruited 20 HIV-infected patients to detect the levels of USP3 and A3G in peripheral blood and analysed their correlation with CD4+ T-cell counts. Correlation was estimated by Pearson correlation coefficients (for parametric data). Results:. The results demonstrated that USP3 specifically inhibits HIV-1 replication in an A3G-dependent manner. Further investigation found that USP3 stabilized 90% to 95% of A3G expression by deubiquitinating Vif-mediated polyubiquitination and blocking its degradation in an enzyme-dependent manner. It also enhances the A3G messenger RNA (mRNA) level by binding to A3G mRNA and stabilizing it in an enzyme-independent manner. Moreover, USP3 expression was positively correlated with A3G expression (r = 0.5110) and CD4+ T-cell counts (r = 0.5083) in HIV-1-infected patients. Conclusions:. USP3 restricts HIV-1 viral infections by increasing the expression of the antiviral factor A3G. Therefore, USP3 may be an important target for drug development and serve as a novel therapeutic strategy against viral infections

Antimalarial and Structural Studies of Pyridine-Containing Inhibitors of 1‑Deoxyxylulose-5-phosphate Reductoisomerase

1-Deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) in the nonmevalonate isoprene biosynthesis pathway is a target for developing antimalarial drugs. Fosmidomycin, a potent DXR inhibitor, showed safety as well as efficacy against <i>Plasmodium falciparum</i> malaria in clinical trials. On the basis of our previous quantitative structure–activity relationship (QSAR) and crystallographic studies, several novel pyridine-containing fosmidomycin derivatives were designed, synthesized, and found to be highly potent inhibitors of <i>P. falciparum</i> DXR (<i>Pf</i>DXR) having <i>K</i>_i values of 1.9–13 nM, with the best one being ∼11× more active than fosmidomycin. These compounds also potently block the proliferation of multidrug resistant <i>P. falciparum</i> with EC₅₀ values as low as 170 nM. A 2.3 Å crystal structure of <i>Pf</i>DXR in complex with one of the inhibitors is reported, showing that the flexible loop of the protein undergoes conformational changes upon ligand binding and a hydrogen bond and favorable hydrophobic interactions between the pyridine group and the <i>Pf</i>DXR account for the enhanced activity