PAS domain containing regulator SLCG_7083 involved in morphological development and glucose utilization in Streptomyces lincolnensis.

BACKGROUND: Streptomyces lincolnensis is well known for producing the clinically important antimicrobial agent lincomycin. The synthetic and regulatory mechanisms on lincomycin biosynthesis have been deeply explored in recent years. However, the regulation involved in primary metabolism have not been fully addressed. RESULTS: SLCG_7083 protein contains a Per-Arnt-Sim (PAS) domain at the N-terminus, whose homologous proteins are highly distributed in Streptomyces. The inactivation of the SLCG_7083 gene indicated that SLCG_7083 promotes glucose utilization, slows mycelial growth and affects sporulation in S. lincolnensis. Comparative transcriptomic analysis further revealed that SLCG_7083 represses eight genes involved in sporulation, cell division and lipid metabolism, and activates two genes involved in carbon metabolism. CONCLUSIONS: SLCG_7083 is a PAS domain-containing regulator on morphological development and glucose utilization in S. lincolnensis. Our results first revealed the regulatory function of SLCG_7083, and shed new light on the transcriptional effects of SLCG_7083-like family proteins in Streptomyces.

Deciphering the photolysis products and biological concerns of triclosan under UVC and UVA.

Triclosan (TCS) is omnipresent in the environment and has drawn increasing attention due to its potential adverse effects on human health. Direct photolysis of TCS readily occurs, especially in the surface layers of waters that receive abundant ultraviolet radiation during the daytime. However, biological concerns and the identification of toxic products during TCS photolysis have been explored limitedly. Therefore, in the present work, the structural characterization of the photolysis products by UVC and UVA were performed based on the mass spectra and fragmental ions. The results displayed that TCS was more readily eliminated by UVC than UVA, and the product species were completely different when TCS was degraded by UVC and UVA, respectively. Two products, m/z 235 and m/z 252, were produced via reductive dechlorination and nucleophilic substitution with UVC, while three dioxin-like isomer products were generated by dechlorination, cyclization and hydroxylation. Furthermore, the results of biological concerns suggested that the elimination of TCS did not represent the disappearance of biological risks. Specifically, more hazardous and photolysis products were formed during TCS photolysis with ultraviolets. For instance, the dioxin-like isomer products were highly microtoxic and genotoxic, and mildly antiestrogenic. The positive findings highlighted the biological concerns of TCS photolysis by ultraviolet radiation in the aquatic environment.

Simultaneous Removal of Trivalent Chromium and Hexavalent Chromium from Soil Using a Modified Bipolar Membrane Electrodialysis System.

In this study, a modified bipolar membrane electrodialysis system equipped with a "back-to-back" soil compartment was fabricated for simultaneous removal of trivalent chromium (Cr(III)) and hexavalent chromium (Cr(VI)) from contaminated soils. The results showed that the soil solution pH had a significant effect on the Cr(III) and Cr(VI) desorption, and the desorption data fit well with the Elovich kinetic model. Current density had an obvious effect on Cr(III) and Cr(VI) removal, cell voltage, soil pH, current efficiency, and specific energy consumption, and the optimal current density was 2.0 mA/cm2. The removal efficiencies of Cr(III) and Cr(VI) were both 99.8%, while Cr(III) and Cr(VI) recoveries were somewhat lower at 87 and 90%, respectively, because some Cr(III) and Cr(VI) were adsorbed by the membranes. An energy consumption analysis indicates that the back-to-back soil compartment equipped system increased the current efficiency and decreased the specific energy consumption. When a system equipped with two back-to-back soil compartments was used to remove chromium from soil, the current efficiency increased to 28.8% and the specific energy consumption decreased to 0.048 kWh/g. The experimental results indicate that the proposed process has the potential to be an effective technique for the treatment of soil contaminated with heavy metals.

Ciprofloxacin degradation performances and mechanisms by the heterogeneous electro-Fenton with flocculated fermentation biochar.

Antibiotic fermentation residue flocculated by polymeric ferric sulfate (PFS) has been classified as a "hazardous waste" in China. In this study, it was recycled into antibiotic fermentation residue biochar (AFRB) by pyrolysis and used as a heterogeneous electro-Fenton (EF) catalyst for ciprofloxacin (CIP) degradation. The results show that PFS was reduced to Fe0 and FeS during pyrolysis, which was beneficial for the EF process. The AFRB with mesoporous structures exhibited soft magnetic features, which were convenient for separation. CIP was completely degraded within 10 min by the AFRB-EF process at an initial concentration of 20 mg/L. Increasing the working current and catalyst dosage within a certain range could improve the degradation rate. ·OH and O2·- were the dominant reactive oxygen species that played critical roles for CIP degradation. The antibacterial groups of CIP have been destroyed by the heterogeneous electro-Fenton process and its toxicity was negligible. The AFRB showed satisfactory performance, even though it was recycled five times. This study provide new insights into the resourceful treatment of antibiotic fermentation residues.

Direct photolysis of diclofenac under simulated sunlight: Transformation pathway and biological concerns.

Topical diclofenac gels are frequently applied on human skin and, consequently are exposed to sunlight during outdoor activities. The degradation of diclofenac (DCF) with sunlight exposure is known to occur but the detailed transformation characteristics and biological concerns have not been comprehensively investigated. In the present work, the transformation products during diclofenac photolysis were identified with the aid of ultra-performance liquid chromatography coupled with triple time-of-flight mass spectrometry (UPLC-TripleTOF). Biological concerns, including microtoxicity, genotoxicity, cytotoxicity and antiestrogenicity were examined with multiple in-vitro bioassays. Spearman correlation analysis was conducted to obtain further insight into the contributions of photolysis products to overall biological concerns. The results demonstrated that diclofenac was readily degraded under sunlight to form five main photolysis products via substitution, dechlorination, dehydroxylation, homodimerization and heterodimerization. Products P1, P2 and P5 were reported previously, while two dimer products (P3 and P4) are innovative products and have not been found in prior studies. A significant elevation in the microtoxicity was found during the photolysis of diclofenac, resulting mainly from the carbazole-containing photolysis products P2, P3, P4 and P5. Genotoxicity and antiestrogenicity declined along with the reduction of diclofenac, indicating that no photolysis products were genotoxic or anti-estrogenic. Modest cytotoxicity to the human skin epidermis cell line was observed and attributed to the formation of intermediate species. This outcome highlighted the biological concerns of diclofenac to human health when exposed to sunlight.

The drinking water disinfection performances and mechanisms of UVA-LEDs promoted by electrolysis.

In this study, the UVA (Ultraviolet A) drinking water disinfection was promoted by electrolysis. The influences of the UVA, electrolysis current, bubbling and temperature were investigated. The disinfection mechanisms and bacterial reactivation had been studied. The results revealed that the treatment time needed to reach the DL (detection limit, about 5.4 log removal) was shortened from 180 to 80 min by the electrolysis. The total electricity consumption decreased from about 126-57.0 kJ/L. Compared with increasing the UVA irradiation, increasing the electrolysis current in a certain range was more preferred to improve the disinfection rate. Oxygen bubbling or higher temperature could enhance the E. coli inactivation. The quenching experiment and EPR (Electron paramagnetic resonance) detection confirmed that ROSs (1O2, ·O2- and ·OH) played important roles for the disinfection. Compared with the treatment with UVA alone, the cell membrane damage was more severe by the promoting method. In addition to the dramatically reduced enzyme activity, the synergistic process degraded most of the bacterial genomic DNA, and the bacteria were completely killed. Therefore, hybrid with electrolysis is a better way for the application of the UVA-LED disinfection.

Spontaneous nutrient recovery and disinfection of aquaculture wastewater via Mg-coconut shell carbon composites.

Aquaculture wastewater contained large amounts of pathogenic microorganisms, nitrogen (N) and phosphorus (P). In this study, the nutrient recoveries and wastewater disinfection were simultaneously achieved using Mg-coconut shell carbon (Mg-CSC). The composites were prepared by a ball milling method. The hydrogen peroxide (H2O2) was in-situ generated by the dissolved oxygen reduction driven by Mg corrosion on the CSC surface, which inactivated the microorganisms. Besides that, Mg corrosion provided sufficient Mg ions and appropriate pH conditions for struvite formation. The results show that 5.4-log E.coli removal was achieved under different conditions. Improving the Mg/CSC ratio and composite dosage could shorten the time required for disinfection. In addition to H2O2, singlet oxygen played a critical role. Reactive oxygen species destroyed the cellular structure and killed the bacteria. The recoveries of NH4+-Nand P under certain conditions were about 60% and 91%, respectively. An increased composite dosage could improve the recovery ratio of P. Excessive dosages were not beneficial for removing NH4+-N. The characterization result revealed that struvite crystals were the main precipitates on the CSC surface. The Mg-CSC composites also revealed satisfied nutrient recovery and disinfection performances in the real aquaculture wastewater treatment process.

Water disinfection by the UVA/electro-Fenton process under near neutral conditions: Performance and mechanisms.

An efficient and thorough water disinfection is critical for human health. In this study, UVA-LEDs, nitrilotriacetic acid (NTA) and a boron-doped diamond anode were respectively used as the UVA source, the iron chelator and the anode for the UVA/electro-Fenton (E-Fenton) reaction to treat wastewater. The disinfection performance of the UVA/E-Fenton had been investigated. The mechanisms of the E. coli inactivation had been clarified. The results showed that complete disinfection (about 5.6-log removal) could be achieved within 50 min at a certain condition due to the synergistic effort of the UVA, anodic oxidation and the electro-Fenton. The quenching experiments and the electron paramagnetic resonance (EPR) detection indicated that •OH, •O2- and 1O2 play important roles for inactivating E. coli. The results of SEM images and genomic DNA electrophoresis suggested that both the cell structure and the DNA had been thoroughly destroyed during the UVA/E-Fenton process. Increasing the UVA irradiation, oxygen bubbling could improve the disinfection rate, while it also would increase the energy consumption. The appropriate Fe and NTA ratio was 1:2 to realize an efficient Fenton reaction under near neutral condition. Complete disinfection was also achieved within 50 min when it used for treating real wastewater. Thus, the UVA/E-Fenton process is a satisfied way for water disinfection.

Electrochemical disinfection using a modified reticulated vitreous carbon cathode for drinking water treatment.

A reticulated vitreous carbon (RVC) cathode modified by anodic polarization in 20 wt% H2SO4 solution was used for drinking water disinfection under a neutral low electrolyte concentration (0.25 g/L Na2SO4) condition. The contribution of the modified RVC anode and the Ti/RuO2 cathode to disinfection was investigated. The influences of current, initial Escherichia coli load, temperature and water volume were studied. The results show that H2O2 generation increased to approximately three times using the modification of the RVC. E. coli was mainly deactivated by the H2O2 generated at the cathode. For water with about 106 CFU/mL E. coli, the detection limit (<4 CFU/mL) was reached under different conditions. Increasing current could simultaneously shorten the treatment time and increase the energy consumption (EC) simultaneously. Although decreasing the initial load reduced the treatment time, the EC for per log E. coli removal increased. The time required for disinfection shortened from 3.5 to 2.5 h and the EC for per log removal decreased from 218.5 to 123.2 Wh/m3 when the temperature increased from 20 to 40 °C. Although more time was required for disinfection, the EC decreased from 218.5 to 141.4 Wh/m3 when the volume was doubled.

Keratin 8 reduces colonic permeability and maintains gut microbiota homeostasis, protecting against colitis and colitis-associated tumorigenesis.

Keratin 8 (CK8) is the major component of the intermediate filaments of simple or single-layered epithelia. Gene targeting mice model suggest that CK8 is involved in colonic active ion transport, colorectal hyperplasia and inflammation. In the present study, we found that CK8 is downregulated in the colon during DSS-induced colitis and AOM/DSS-induced colitis-associated colorectal cancer (CAC) development. In human patients with colon cancer, CK8 is downregulated. Using CK8 heterozygous knockout mice (CK8+/-), we found that CK8+/- mice are highly susceptible to DSS-induced colitis and more prone to AOM/DSS-induced CAC than wild type (WT) mice. The colonic permeability is increased with DSS or AOM/DSS treatment, leading to alteration of gut microbiota in CK8+/- mice with CAC. Metagenomic analysis of fecal microbiota suggests Firmicutes and Proteobacteria are increased in CK8+/- mice with CAC, while Bacteroidetes and Verrucomicrobia are decreased. Antibiotic treatment decreases the incidence of colorectal cancer tumorigenesis and TLR4 inhibitor attenuates the susceptibility of CK8+/- mice to DSS-induced colitis. These data suggest CK8 protects mice from colitis and colitis-associated colorectal cancer by modulating colonic permeability and gut microbiota composition homeostasis.

Keratin 8 limits TLR-triggered inflammatory responses through inhibiting TRAF6 polyubiquitination.

Toll-like receptors (TLRs) have critical roles in innate immunity and inflammation and the detailed mechanisms by which TLR signaling is fine tuned remain unclear. Keratin 8 (CK8) belongs to the type II keratin family and is the major compontent of the intermediate filaments of simple or single-layered epithelia. Here we report that down-regulation of CK8 in mice enhanced TLR-mediated responses, rendering mice more susceptible to lipopolysaccharide (LPS)-induced endotoxin shock and Escherichia coli-caused septic peritonitis with reduced survival, elevated levels of inflammation cytokines and more severe tissue damage. We found that CK8 suppressed TLR-induced nuclear factor (NF)-κB activation and interacted with the adaptor tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) to prevent its polyubiquitination. Our findings demonstrate a novel role of CK8 in negative regulation of TLR/NF-κB signaling and highlight a previously unidentified nonclassical function for CK8 in limiting inflammatory responses.

[Construction of a GFP/Puro double-labeled lentiviral vector containing CK8 interfering RNA and its effect on cell apoptosis in vitro].

OBJECTIVE: To construct a GFP/Puro double-labeled lentiviral expression vector for CK8 silencing and assess the effects of CK8 silencing on cell apoptosis. METHODS: The siRNA sequences of CK8 were inserted into the lentiviral expression vector GV248 and transfected into 293T cells with the packaging plasmids PMD and SPA. The lentivirus was collected at 24 and 36 h post-transfection. Flow cytometry was used to detect the virus titer and the positive cells were selected with puromycin. The knockdown of CK8 was examined by Western blotting. The effect of CK8 down-regulation on cell apoptosis induced by cisplatin was detected with Annexin V/PI staining. RESULTS AND CONCLUSION: We successfully constructed CK8 interference lentiviral vector and obtained a stable cell line with CK8 knock-down that was sensitive to cisplatin-induced apoptosis.