Systematic strategies for developing phage resistant Escherichia coli strains.

Phages are regarded as powerful antagonists of bacteria, especially in industrial fermentation processes involving bacteria. While bacteria have developed various defense mechanisms, most of which are effective against a narrow range of phages and consequently exert limited protection from phage infection. Here, we report a strategy for developing phage-resistant Escherichia coli strains through the simultaneous genomic integration of a DNA phosphorothioation-based Ssp defense module and mutations of components essential for the phage life cycle. The engineered E. coli strains show strong resistance against diverse phages tested without affecting cell growth. Additionally, the resultant engineered phage-resistant strains maintain the capabilities of producing example recombinant proteins, D-amino acid oxidase and coronavirus-encoded nonstructural protein nsp8, even under high levels of phage cocktail challenge. The strategy reported here will be useful for developing engineered E. coli strains with improved phage resistance for various industrial fermentation processes for producing recombinant proteins and chemicals of interest.

Corrigendum: Autophagy Blockade by Ai Du Qing Formula Promotes Chemosensitivity of Breast Cancer Stem Cells Via GRP78/ß-Catenin/ABCG2 Axis.

[This corrects the article DOI: 10.3389/fphar.2021.659297.].

What is the general Chinese public's awareness of and attitudes towards Helicobacter pylori screening and associated health behaviours? A cross-sectional study.

OBJECTIVE: To evaluate the general population's awareness of and attitudes toward Helicobacter pylori (HP) screening and health behaviours. DESIGN: Cross-sectional study. SETTING: Hengyang, Hunan Province, China. PARTICIPANTS: Using stratified cluster random sampling, a pretested structured questionnaire was used to interview members of the general population aged ≥18 years. PRIMARY AND SECONDARY OUTCOME MEASURES: Knowledge of and attitudes toward HP screening and associated health behaviours, sociodemographic factors associated with HP knowledge, and screening behaviours. RESULTS: This study featured 1042 participants. The average knowledge score was 11 (QL=4, QU=20, range 0-29). Approximately 68.9% of the participants said they had heard of HP, but 67.5% had never had an HP test. The most common reasons for not undergoing screening were 'no symptoms' (55.7%) and 'lack of knowledge regarding the benefits of the test' (21.1%). Independent factors related to knowledge included age, education level, occupation, HP infection, frequency of drinking unboiled water (p<0.05). Factors independently associated with screening behaviour included occupation, average monthly income, presence/absence of indigestion, stomach discomfort or pain, and/or stomach disease and knowledge score (p<0.05). Overall, 941 (90.3%) participants never used anti-HP toothpaste, and 442 (40.5%) never used serving spoons or chopsticks. The risk factors for HP infection included eating out and eating in groups (p<0.05). CONCLUSION: In China, the general population has poor knowledge of HP, but most people have a positive attitude towards HP screening. Being asymptomatic and lacking knowledge about testing were the main reasons for reluctance to be screened. These results highlight the urgent need for educational activities to raise awareness, enhance screening rates for HP, and encourage people to adopt a healthy lifestyle.

Source contribution analysis of PM2.5 using Response Surface Model and Particulate Source Apportionment Technology over the PRD region, China.

Identifying the emission source contributions to PM2.5 is essential for a sound PM2.5 pollution control policy. In this study, we conduct a comparative analysis of PM2.5 source contributions over the Pearl River Delta (PRD) region of China using two advanced source contribution modeling techniques: Response Surface Model (RSM) and Particulate Source Apportionment Technology (PSAT). Our comparative analyses show that RSM and PSAT can both reasonably predict the contribution of primary PM2.5 emission sources to PM2.5 formation due to its linear nature. For the secondary PM2.5 formed by the nonlinear reactions among PM2.5 precursors, however, our study shows that PSAT appears to have limitations in quantifying the nonlinear contribution of PM2.5 precursors to emission reductions, while RSM seems to better address the nonlinear relationship among PM2.5 precursors (e.g., PM2.5 disbenefits due to local NOx emission reductions in major cities with high NOx emissions). The pilot study case results show that for the ambient PM2.5 in the central cities (Guangzhou, Shenzhen, Foshan, Dongguan, and Zhongshan) of the PRD, the regional source emissions contribute the most by 42-66%; the dust emissions are the top contribution sources (29-34% by RSM and 27-31% by PSAT), and the mobile sources are listed as the secondary contributors accounting for 16-25% by RSM and 19-30% by PSAT among the anthropogenic emission sources. The city-scale cooperation on emission reductions and the enhancement of dust and mobile emission control are recommended to effectively reduce the ambient PM2.5 concentration in the PRD.

Awareness, attitude and barriers of colorectal cancer screening among high-risk populations in China: a cross-sectional study.

OBJECTIVE: To assess the awareness, attitude and barriers of colorectal cancer screening among high-risk populations in China. DESIGN: A cross-sectional study was employed. SETTING: This study was conducted in nine hospitals in Hunan province, China. PARTICIPANTS: Individuals with a high-risk for colorectal cancer were interviewed using a pretested structured questionnaire. PRIMARY AND SECONDARY OUTCOME MEASURES: Knowledge, attitude towards colorectal cancer screening, sociodemographic factors associated with screening knowledge and behaviour and barriers of colorectal cancer screening. RESULTS: This study included 684 participants. The mean knowledge score was 11.86/24 (SD 4.84). But over 70% of them held a positive attitude towards screening. Only 13.3% had undergone colorectal cancer screening. Independent factors related to knowledge were education level of college or above, working as a white collar, higher income, having health insurance, having seen a doctor in the past year and with a high perceived risk (p<0.05). Factors independently associated with screening behaviour included personal history of colorectal disease, having seen a doctor in the past year, previous discussion of colorectal cancer screening, high perceived risk and better knowledge (p<0.05). Main reasons for not undergoing screening were no symptoms or discomfort (71.1%), never having thought of the disease or screening (67.4%) and no doctor advised me (29.8%). CONCLUSION: In China, the majority of high-risk people had deficient knowledge and had never undergone colorectal cancer screening. But most of them held a positive attitude towards the benefits of colorectal cancer screening. This has promising implications to design targeted educational campaigns and establish screening programmes to improve colorectal cancer awareness and screening participation. Healthcare professionals should advise high-risk individuals to participate in screening and inform them about cancer risk.

Source impact and contribution analysis of ambient ozone using multi-modeling approaches over the Pearl River Delta region, China.

Quantification of source impacts and contributions is a key element for the design of effective air pollution control policies. In this study, O3 source impacts and contributions were comprehensively assessed over the Pearl River Delta (PRD) region of China using brute-force method (BFM), response surface modeling with BFM (RSM-BFM) and differential method (RSM-DM) respectively, high-order decoupled direct method (HDDM), and ozone source apportionment technology (OSAT). The multi-modeling comparison results indicated that under typical nonlinear atmospheric conditions during the O3 formation, BFM, RSM-BFM, and HDDM seemed to be appropriate for assessing the impact of single source emissions; however, the results of HDDM could deviate from those of BFM when the emission reduction ratio was higher than 50 %. Under multi-source control scenarios, the results of source contribution analyses obtained from RSM-DM and OSAT were reasonably well, but the performance of OSAT was limited by its capability in representing the nonlinearity of O3 response to emission reductions of its precursors, particularly NOx. The results of this pilot study in the PRD showed that the RSM-DM appeared to replicate the nonlinearity of O3 chemistry reasonably well (e.g., O3 disbenefits due to local NOx emission reductions in Guangzhou city). Based on the source contribution results, on-road mobile (including both NOx and VOC emissions) and industrial process (mainly VOC emissions) sources were identified as two major contribution sectors by both RSM-DM and OSAT, contributing an average of 31.5 % and 11.4 % (estimated by RSM-DM) and 29.2 % and 13.0 % (estimated by OSAT) respectively to O3 formation in 9 cities of the PRD. Therefore, the reinforced emission reductions on NOx and VOC from on-road mobile and industrial process sources in the central cities (i.e., Guangzhou, Foshan, Dongguan, Shenzhen, and Zhongshan) were suggested to effectively mitigate the ambient O3 levels in the PRD.

Autophagy Blockade by Ai Du Qing Formula Promotes Chemosensitivity of Breast Cancer Stem Cells Via GRP78/ß-Catenin/ABCG2 Axis.

Accumulating evidence suggests that the root of drug chemoresistance in breast cancer is tightly associated with subpopulations of cancer stem cells (CSCs), whose activation is largely dependent on taxol-promoting autophagy. Our pilot study identified GRP78 as a specific marker for chemoresistance potential of breast CSCs by regulating Wnt/ß-catenin signaling. Ai Du Qing (ADQ) is a traditional Chinese medicine formula that has been utilized in the treatment cancer, particularly during the consolidation phase. In the present study, we investigated the regulatory effects and molecular mechanisms of ADQ in promoting autophagy-related breast cancer chemosensitivity. ADQ with taxol decreasing the cell proliferation and colony formation of breast cancer cells, which was accompanied by suppressed breast CSC ratio, limited self-renewal capability, as well as attenuated multi-differentiation. Furthermore, autophagy in ADQ-treated breast CSCs was blocked by taxol via regulation of ß-catenin/ABCG2 signaling. We also validated that autophagy suppression and chemosensitizing activity of this formula was GRP78-dependent. In addition, GRP78 overexpression promoted autophagy-inducing chemoresistance in breast cancer cells by stabilizing ß-catenin, while ADQ treatment downregulated GRP78, activated the Akt/GSK3ß-mediated proteasome degradation of ß-catenin via ubiquitination activation, and consequently attenuated the chemoresistance-promoted effect of GRP78. In addition, both mouse breast cancer xenograft and zebrafish xenotransplantation models demonstrated that ADQ inhibited mammary tumor growth, and the breast CSC subpopulation showed obscure adverse effects. Collectively, this study not only reveals the chemosensitizating mechanism of ADQ in breast CSCs, but also highlights the importance of GRP78 in mediating autophagy-promoting drug resistance via ß-catenin/ABCG2 signaling.

SspABCD-SspFGH Constitutes a New Type of DNA Phosphorothioate-Based Bacterial Defense System.

Unlike nucleobase modifications in canonical restriction-modification systems, DNA phosphorothioate (PT) epigenetic modification occurs in the DNA sugar-phosphate backbone when the nonbridging oxygen is replaced by sulfur in a double-stranded (ds) or single-stranded (ss) manner governed by DndABCDE or SspABCD, respectively. SspABCD coupled with SspE constitutes a defense barrier in which SspE depends on sequence-specific PT modifications to exert its antiphage activity. Here, we identified a new type of ssDNA PT-based SspABCD-SspFGH defense system capable of providing protection against phages through a mode of action different from that of SspABCD-SspE. We provide further evidence that SspFGH damages non-PT-modified DNA and exerts antiphage activity by suppressing phage DNA replication. Despite their different defense mechanisms, SspFGH and SspE are compatible and pair simultaneously with one SspABCD module, greatly enhancing the protection against phages. Together with the observation that the sspBCD-sspFGH cassette is widely distributed in bacterial genomes, this study highlights the diversity of PT-based defense barriers and expands our knowledge of the arsenal of phage defense mechanisms.IMPORTANCE We recently found that SspABCD, catalyzing single-stranded (ss) DNA phosphorothioate (PT) modification, coupled with SspE provides protection against phage infection. SspE performs both PT-simulated NTPase and DNA-nicking nuclease activities to damage phage DNA, rendering SspA-E a PT-sensing defense system. To our surprise, ssDNA PT modification can also pair with a newly identified 3-gene sspFGH cassette to fend off phage infection with a different mode of action from that of SspE. Interestingly, both SspFGH and SspE can pair with the same SspABCD module for antiphage defense, and their combination provides Escherichia coli JM109 with additive phage resistance up to 105-fold compared to that for either barrier alone. This agrees with our observation that SspFGH and SspE coexist in 36 bacterial genomes, highlighting the diversity of the gene contents and molecular mechanisms of PT-based defense systems.

SspABCD-SspE is a phosphorothioation-sensing bacterial defence system with broad anti-phage activities.

Bacteria have evolved diverse mechanisms to fend off predation by bacteriophages. We previously identified the Dnd system, which uses DndABCDE to insert sulfur into the DNA backbone as a double-stranded phosphorothioate (PT) modification, and DndFGH, a restriction component. Here, we describe an unusual SspABCD-SspE PT system in Vibrio cyclitrophicus, Escherichia coli and Streptomyces yokosukanensis, which has distinct genetic organization, biochemical functions and phenotypic behaviour. SspABCD confers single-stranded and high-frequency PTs with SspB acting as a nickase and possibly introducing nicks to facilitate sulfur incorporation. Strikingly, SspABCD coupled with SspE provides protection against phages in unusual ways: (1) SspE senses sequence-specific PTs by virtue of its PT-stimulated NTPase activity to exert its anti-phage activity, and (2) SspE inhibits phage propagation by introducing nicking damage to impair phage DNA replication. These results not only expand our knowledge about the diversity and functions of DNA PT modification but also enhance our understanding of the known arsenal of defence systems.

Awareness of risk factors and warning symptoms and attitude towards gastric cancer screening among the general public in China: a cross-sectional study.

OBJECTIVES: This study aimed to assess the knowledge of risk factors and warning symptoms and attitude towards gastric cancer screening among the general population in China. SETTING: Hunan province, China PARTICIPANTS: Individuals aged older than 18 years were recruited using a cluster sampling method. DESIGN: A cross-sectional study, and a pretested structured questionnaire was used to assess participants' awareness of gastric cancer. PRIMARY AND SECONDARY OUTCOME MEASURES: Knowledge level of risk factors and warning symptoms of gastric cancer, gastric cancer screening attitude, sociodemographic factors associated with gastric cancer knowledge and screening behaviour. RESULTS: This study comprised 1200 participants with a mean age of 40.31 (SD 16.73) years, of whom 622 (51.8%) were women. The mean score for gastric cancer knowledge was 8.85/22 (SD 6.48). There were 47.0% of the participants who had a low knowledge level about the risk factors and warning symptoms of gastric cancer. In total, 83.8% believed screening is helpful for early detection of gastric cancer, and 15.2% had undergone gastric cancer screening. The most common reason for not undergoing screening was having 'no symptoms' (63.0%), followed by 'fear of undergoing gastroscopy' (38.1%). Independent factors related to lower knowledge levels included male sex, living in rural areas, lower educational level, working as a farmer and without a family history of gastric cancer (p<0.05). Factors independently associated with screening behaviour included white-collar employment, higher income and having upper gastrointestinal tract diseases (p<0.05). CONCLUSIONS: In China, people have poor knowledge about risk factors and warning symptoms of gastric cancer, but a majority have a positive attitude towards the benefits of gastric cancer screening. Being asymptomatic and having a fear of gastroscopy were the main self-reported reasons for not undergoing screening. These results highlight the urgent need for educational campaigns to improve gastric cancer awareness.