Modulation of Type III Secretion System in Pseudomonas aeruginosa: Involvement of the PA4857 Gene Product

Pseudomonas aeruginosais an opportunistic pathogen that causes serious acute or chronic infections in humans.Acute infections typically involve the type Ш secretion systems (T3SS) and bacterial motility,whereas chronic infectionsare often associated with biofilm formation and the type VI secretion system (T6SS). To identifynew genes required for pathogenesis, a transposon mutagenesis library was constructed and the gene PA4857, named tspR, was found to modulateT3SS gene expression. Deletion of P. aeruginosa tspRreduced the virulence in a mouse acute lung infection model and diminished cytotoxicity. Suppression of T3SS gene expression in the tspR mutant resulted from compromised translation of the T3SS master regulator ExsA. TspR negatively regulated two small RNAs, RsmYand RsmZ, which control RsmA. Our data demonstrated that defects inT3SS expression and biofilm formation in retS mutant could be partially restored by overexpression of tspR. Taken together, our results demonstrated thatthe newly identifiedretS-tspRpathway is coordinated with the retS-gacSsystem, which regulates the genes associated with acute and chronic infections andcontrols the lifestyle choice of P. aeruginosa

A Unique ATPase, ArtR (PA4595), Represses the Type III Secretion System in Pseudomonas aeruginosa

Pseudomonas aeruginosa is an important human pathogen which uses the type III secretion system (T3SS) as a primary virulence factor to establish infections in humans. The results presented in this report revealed that the ATP-binding protein PA4595 (named ArtR, a Regulator that is an ATP-activated Repressor of T3SS) represses T3SS expression in P. aeruginosa. The expression of T3SS genes, including exoS, exoY, exoT, exsCEBA, and exsD-pscB-L, increased significantly when artR was knockout. The effect of ArtR on ExsA is at the transcriptional level, not at the translational level. The regulatory role and cytoplasm localization of ArtR suggest it belongs to the REG sub-family of ATP-binding cassette (ABC) family. Purified GST-tagged ArtR showed ATPase activity in vitro. The conserved aspartate residues in the dual Walker B motifs prove to be essential for the regulatory function of ArtR. The regulation of T3SS by ArtR is unique, which does not involve the known GacS/A-RsmY/Z-RsmA-ExsA pathway or Vfr. This is the first REG subfamily of ATP-binding cassette that is reported to regulate T3SS genes in bacteria. The results specify a novel player in the regulatory networks of T3SS in P. aeruginosa

Glutathione Activates Type III Secretion System Through Vfr in Pseudomonas aeruginosa

Glutathione (GSH) is the most abundant antioxidant in all living organisms. Previously, we have shown that a deletion mutant in the glutathione synthetase gene (ΔgshB) decreases the expression of type III secretion system (T3SS) genes of Pseudomonas aeruginosa. However, the mechanism remains elusive. In this study, a comprehensive transcriptomic analysis of the GSH-deficient mutant ΔgshAΔgshB was used to elucidate the role of GSH in the pathogenesis of P. aeruginosa. The data show that the expression of genes in T3SS, type VI secretion system (T6SS) and some regulatory genes were impaired. ΔgshAΔgshB was attenuated in a mouse model of acute pneumonia, swimming and swarming motilities, and biofilm formation. Under T3SS inducing conditions, GSH enhanced the expression of T3SS in both wild-type PAO1 and ΔgshAΔgshB, but not in Δvfr. Genetic complementation of Δvfr restored the ability of GSH to induce the expression of T3SS genes. Site-directed mutagenesis based substitution of cysteine residues with alanine in Vfr protein abolished the induction of T3SS genes by GSH, confirming that GSH regulates T3SS genes through Vfr. Exposure to H2O2 decreased free thiol content on Vfr, indicating that the protein was sensitive to redox modification. Importantly, GSH restored the oxidized Vfr to reduced state. Collectively, these results suggest that GSH serves as an intracellular redox signal sensed by Vfr to upregulate T3SS expression in P. aeruginosa. Our work provides new insights into the role of GSH in P. aeruginosa pathogenesis

Sodium Selenite Enhances Antibiotics Sensitivity of Pseudomonas aeruginosa and Deceases Its Pathogenicity by Inducing Oxidative Stress and Inhibiting Quorum Sensing System

Pseudomonas aeruginosa, a Gram-negative opportunistic pathogen, is commonly found in clinical settings and immuno-compromised patients. It is difficult to be eradicated due to its strong antibiotic resistance, and novel inactivation strategies have yet to be developed. Selenium is an essential microelement for humans and has been widely used in dietary supplement and chemoprevention therapy. In this study, the physiological and biochemical effects of sodium selenite on P. aeruginosa PAO1 were investigated. The results showed that 0~5 mM sodium selenite did not impact the growth of PAO1, but increased the lethality rate of PAO1 with antibiotics or H2O2 treatment and the antibiotics susceptibility both in planktonic and biofilm states. In addition, sodium selenite significantly reduced the expression of quorum sensing genes and inhibited various virulence factors of this bacterium, including pyocyanin production, bacterial motilities, and the type III secretion system. Further investigation found that the content of ROS in cells was significantly increased and the expression levels of most genes involved in oxidative stress were up-regulated, which indicated that sodium selenite induced oxidative stress. The RNA-seq result confirmed the phenotypes of virulence attenuation and the expression of quorum sensing and antioxidant-related genes. The assays of Chinese cabbage and Drosophila melanogaster infection models showed that the combination of sodium selenite and antibiotics significantly alleviated the infection of PAO1. In summary, the results revealed that sodium selenite induced oxidative stress and inhibited the quorum sensing system of P. aeruginosa, which in turn enhanced the antibiotic susceptibility and decreased the pathogenicity of this bacterium. These findings suggest that sodium selenite may be used as an effective strategy for adjunct treatment of the infections caused by P. aeruginosa

The shattered 'Iron Rice Bowl': Intergenerational effects of Chinese State-Owned Enterprise reform

Reform of the Chinese State-Owned Enterprise (SOE) sector in the late 1990s triggered massive layoffs (34 million employees) and marked the end of the “Iron Rice Bowl” guarantee of employment security for the remaining 67 million workers. An expanding international literature has documented the adverse health impacts of economic insecurity on adults, but has typically neglected children. This paper uses the natural experiment of SOE reform to explore the causal relationship between increased parental economic insecurity and children's BMI Z-score. Using province-year-level layoff rates and income loss from the layoffs, we estimate a generalised difference-in-differences model with child fixed effects and year fixed effects. For a medium-build 10-year-old boy, a median treatment effect implies a gain of 1.8 kg and a 2.2-percentage-point increase in the overweight rate due to the reform. Anxiety about potential losses causes weight gain for boys whose SOE parents kept their jobs. Unconditional quantile regressions suggest that boys who are heavier are more likely to gain weight. Girls are not significantly affected. Intergenerational effects therefore increase the estimated public health costs of greater economic insecurity

A GM-Based Energy Management Strategy of Hybrid Power System for Hydrogen Fuel Cell Buses

Hydrogen energy is a clean, carbon-free, flexible, efficient, and widely used secondary energy source, which is an ideal alternative to promote the clean and efficient use of traditional fossil fuels. Hydrogen fuel cell bus has the advantages of a high-energy conversion rate, absolute pollution-free, sufficient raw materials, and convenient filling. The hybrid power system, composed of fuel cell and auxiliary energy source, is one of the key technologies to promote the development of hydrogen fuel cell vehicle. This study aims to propose an energy management strategy by analyzing the output characteristics and power allocation of fuel cell and power battery in the hybrid power mode with fuel cell as the main and battery as the auxiliary. A GM (1, N) power prediction strategy was proposed and compared with other strategies as an on-off control strategy and logical threshold value strategy in this study. The variation curves of the battery SOC and fuel cell output power under two working conditions of CCBC and real vehicle conditions were analyzed by using these three strategies, when the initial SOC of power battery is 30%, 70%, and 90%, respectively. Results showed that the power prediction strategy based on GM (1, N) has a better performance in output efficiency and fuel economy when compared to the other two strategies by analyzing the aspects of the battery in the SOC variation and equivalent hydrogen consumption and the fuel cell in the output power variation and hydrogen consumption. This research can be helpful to provide the suggested solution for energy management of the hybrid power system for hydrogen fuel cell buses

Ginkgo biloba extract-loaded PLGA microcapsules generated from microfluidics for Alzheimer's disease treatment

Boosting the birth rate of new neurons in the adult hippocampus is a possible treatment method for the management of Alzheimer's disease (AD). Recent studies have revealed that Ginkgo biloba extract (GBE) can act as a neuroprotective agent to prevent memory loss in a variety of neurodegenerative illnesses. However, the low bioavailability and short half-life of active components in vivo limit their application. Here, we describe a class of microcapsules (MPs) for sustained GBE delivery that have gelatin methacrylate (GelMa) cores and poly(L-lactide-co-glycolide) (PLGA) shells. The encapsulated GBE was slowly released from the solidified MPs during degradation of the polymer. In addition, the MPs showed good biocompatibility in cell experiments. When GelMa-PLGA MPs containing GBE were applied to APP/PS1 mice, amyloid deposition in the mouse brain was reduced, and cognitive impairment was improved. These results indicate that GBE-encapsulated MPs have potential clinical value in treating AD and other neurodegenerative diseases