Staphylococcus aureus can cause different types of diseases from mild skin infections to life-threatening sepsis worldwide. Owing to the emergence and transmission of multidrug-resistant strains, developing an impactful immunotherapy especially vaccine control approach against S. aureus infections is increasingly encouraged and supported. S. aureus manganese transport protein C (MntC), which is a highly-conserved cell surface protein, can elicit protective immunity against S. aureus and Staphylococcus epidermidis. In this study, we evaluated the humoral immune response and CD4+ T cell-mediated immune responses in a mouse peritonitis model. The results showed that MntC-specific antibodies conferred an essential protection for mice to reduce invasion of S. aureus, which was corroborated via the opsonophagocytic killing assay and passive immunization experiment in mice, and moreover MntC-induced Th17 played a remarkable part in preventing S. aureus infection since the MntC-induced protective immunity decreased after neutralization of IL-17 by antibody in vivo and the Th17 adoptive transferred-mice could partly resist S. aureus challenge. In conclusion, we considered that the MntC-specific antibodies and MntC-specific Th17 cells play cooperative roles in the prevention of S. aureus infection.

Pot experiments were conducted to investigate the impact of biochar loading level on soybean growth and physico-chemical properties of alkaline soil. Biochar derived from corn straw was mixed with alkaline soil at 0%, 2.5%, 5%, and 10% loading levels and exposed to the natural elements. Soybean was used as the test crop. The results indicated that a single application of biochar positively and significantly improved soybean productivity and quality attributes of the tested alkaline soil. Soybean yield peaked at 5% loading level, but it declined at 10% loading. Applications of biochar at 5% and 10% loading significantly increased total soil porosity by 4.14% and 5.09%, and decreased the soil pH value by 0.07 and 0.24 units, respectively. Biochar addition significantly increased water holding capacity, total organic carbon content, total nitrogen, Olsen-P, available potassium, and cation exchange capacity. The results indicated that applications of corn straw biochar to alkaline soil improved soybean growth and promoted the physico-chemical properties of alkaline soil. However, the negative effects of increased C:N ratios and soil exchange sodium percentages at higher biochar loading levels should be taken into account when applying biochar as amendments to alkaline soils.

ATP-binding cassette transporters are ubiquitous in almost all organisms. The Escherichia coli genome is predicted to encode 69 ABC transporters. Eleven of the ABC transporters are presumed to be exporters, of which seven are possible drug export transporters. There has been minimal research on the function of YbhFSR, which is one of the putative drug resistance exporters. In this study, the ybhF gene of this transporter was characterized. Overexpression and knockout strains of ybhF were constructed. The ATPase activity of YbhF was studied using the malachite green assay, and the efflux abilities of knockout strains were demonstrated by using ethidium bromide (EB) as a substrate. The substrates of YbhFSR efflux, examined with the minimum inhibitory concentration (MIC), were determined to be tetracycline, oxytetracycline, chlortetracycline, doxycycline, EB, and Hoechst33342. Furthermore, tetracycline and EB efflux and accumulation experiments confirmed that the substrates of YbhFSR were tetracyclines and EB. The MIC assay and the fluorescence test results showed that tetracyclines are likely to be the major antibiotic substrate of YbhFSR. The existence of the signature NatA motif suggested that YbhFSR may also function as a Na + /H + transporter. Overexpression of YbhF in E. coli KNabc lacking crucial Na + /H + transporters conferred tolerance to NaCl, LiCl, and an alkaline pH. Together, the results showed that YbhFSR exhibited dual functions as a drug efflux pump and a Na + (Li +)/H + antiporter.

/nodulation/cell division (RND), the proteobacterial antimicrobial compound efflux (PACE) family and the p-aminobenzoyl-glutamate transporter (AbgT) family [1]. Of these, SMR, MFS, MATE, RND, PACE and AbgT use proton kinetic potential as energy and ABC transporters use ATP as energy. All of them can export drugs and other substrates out of cells [2]. Some proteins, although they confer low levels of resistance, are often the first step in resistance, eventually leading to higher resistance by acquiring chromosomal mutations that target antibiotics. The ABC transporter is one of these proteins [3]. ABC transporters exist in almost all organisms from microorganisms to humans [4, 5]. ABC transporters have attracted extensive attention because of their involvement in important physiological processes such as bacterial resistance, human cystic fibrosis, and tumor cell resistance to chemotherapy drugs [6]. ABC transporters contain similar topologies, with two transmembrane domains (TMD) and two nucleotide-binding domains (NBD). These four domains can fuse to form a full transporter such as the multidrug efflux pump P-glycoprotein (P-gp, ABCB1 or MDR1) in humans. In bacteria, multidrug efflux pumps usually fuse to form a half-transporter with a TMD and an NBD domain [7]. The half-transporter forms a homodimer such as MsbA [7] or LmrA [8]. It can also form heterodimers such as EfrCD [9], YheI/YheH [10] and PatA/PatB [11]. The E. coli K-12MG1655 strain genome was predicted to encode 69 ABC transporters. Eleven of the ABC transporters are presumed to be exporters [5, 12]. Efflux systems of the ABC family in E. coli were reported to mainly include capsular polysaccharide transport pump KpsT [13], hemolysin transport pump HlyA [14, 15], macrocyclic lipid drug efflux pump MacAB [16], outer membrane lipoprotein transport pump LolACDE [17-19], multidrug resistance and phospholipid A transporter MsbA [20], micromycin J25 efflux pump YojI [21], and heme efflux pump CcmABC [22]. Drug efflux systems in E. coli that have been reported included MsbA and MacA. MsbA is a multidrug efflux pump and MacAB is a single-drug efflux pump of macrolides[16]. Studies reported on the A putative multidrug efflux gene, yddA, was cloned from the Escherichia coli K-12 strain. A drugsensitive strain of E. coli missing the main multidrug efflux pump AcrB was constructed as a host and the yddA gene was knocked out in wild-type (WT) and drug-sensitive E. coliΔacrB to study the yddA function. Sensitivity to different substrates of WT E.coli, E. coliΔyddA, E. coliΔacrB and E. coliΔacrBΔyddA strains was compared with minimal inhibitory concentration (MIC) assays and fluorescence tests. MIC assay and fluorescence test results showed that YddA protein was a multidrug efflux pump that exported multiple substrates. Three inhibitors, ortho-vanadate, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and reserpine, were used in fluorescence tests. Ortho-vanadate and reserpine significantly inhibited the efflux and increased accumulation of ethidium bromide and norf...

Interferon-γ (IFN-γ), a cytokine produced by activated natural killer cells and T lymphocytes, is an important regulator of innate and adaptive immunity. Interleukin (IL)-18, also known as IFN-γ-inducing factor, is a cytokine that induces T and natural killer cells to produce IFN-γ. In this study, the chicken IL-18 (ChIL-18) and chicken IFN-γ (ChIFN-γ) genes were inserted into the pET28a prokaryotic expression vector, resulting in pET28a-IL-18 and pET28a-IFN-γ, respectively. These plasmids were transformed into Escherichia coli strain BL21, and the ChIL-18 and ChIFN-γ proteins were expressed and purified. To determine their antiviral activities, 200 ng/mL of ChIL-18 and/or ChIFN-γ were inoculated into chicken embryonic fibroblast cells. After 24 h, one 50% tissue culture infective dose (TCID₅₀) of infectious bursal disease virus (IBDV) was inoculated into the chicken embryonic fibroblast cells. The results showed that the antiviral effect of ChIL-18 and ChIFN-γ in combination was better than that of ChIL-18 or ChIFN-γ alone. Next, 14-day-old chicken were injected with 200 µg of ChIL-18 and/or ChIFN-γ and then were challenged with 10³ TCID₅₀ of IBDV via intraperitoneal injection. The results showed that the proliferation of IBDV was inhibited by the injection of the recombinant proteins, especially the combination of ChIL-18 and ChIFN-γ, as evidenced by cytokine detection, quantitative PCR, and pathology analyses. These results indicate that ChIL-18 and ChIFN-γ could inhibit IBDV infection and the combination of ChIL-18 and ChIFN-γ has a better inhibitory effect than either cytokine alone.