RES-Xre toxin-antitoxin locus knaAT maintains the stability of the virulence plasmid in Klebsiella pneumoniae.

Hypervirulent Klebsiella pneumoniae isolates have been increasingly reported worldwide, especially hypervirulent drug-resistant variants owing to the acquisition of a mobilizable virulence plasmid by a carbapenem-resistant strain. This pLVPK-like mobilizable plasmid encodes various virulence factors; however, information about its genetic stability is lacking. This study aimed to investigate the type II toxin-antitoxin (TA) modules that facilitate the virulence plasmid to remain stable in K. pneumoniae. More than 3,000 TA loci in 2,000 K. pneumoniae plasmids were examined for their relationship with plasmid cargo genes. TA loci from the RES-Xre family were highly correlated with virulence plasmids of hypervirulent K. pneumoniae. Overexpression of the RES toxin KnaT, encoded by the virulence plasmid-carrying RES-Xre locus knaAT, halts the cell growth of K. pneumoniae and E. coli, whereas co-expression of the cognate Xre antitoxin KnaA neutralizes the toxicity of KnaT. knaA and knaT were co-transcribed, representing the characteristics of a type II TA module. The knaAT deletion mutation gradually lost its virulence plasmid in K. pneumoniae, whereas the stability of the plasmid in E. coli was enhanced by adding knaAT, which revealed that the knaAT operon maintained the genetic stability of the large virulence plasmid in K. pneumoniae. String tests and mouse lethality assays subsequently confirmed that a loss of the virulence plasmid resulted in reduced pathogenicity of K. pneumoniae. These findings provide important insights into the role of the RES-Xre TA pair in stabilizing virulence plasmids and disseminating virulence genes in K. pneumoniae.

TADB 3.0: an updated database of bacterial toxin-antitoxin loci and associated mobile genetic elements.

TADB 3.0 (https://bioinfo-mml.sjtu.edu.cn/TADB3/) is an updated database that provides comprehensive information on bacterial types I to VIII toxin-antitoxin (TA) loci. Compared with the previous version, three major improvements are introduced: First, with the aid of text mining and manual curation, it records the details of 536 TA loci with experimental support, including 102, 403, 8, 14, 1, 1, 3 and 4 TA loci of types I to VIII, respectively; Second, by leveraging the upgraded TA prediction tool TAfinder 2.0 with a stringent strategy, TADB 3.0 collects 211 697 putative types I to VIII TA loci predicted in 34 789 completely sequenced prokaryotic genomes, providing researchers with a large-scale dataset for further follow-up analysis and characterization; Third, based on their genomic locations, relationships of 69 019 TA loci and 60 898 mobile genetic elements (MGEs) are visualized by interactive networks accessible through the user-friendly web page. With the recent updates, TADB 3.0 may provide improved in silico support for comprehending the biological roles of TA pairs in prokaryotes and their functional associations with MGEs.

Liquid-Liquid Phase Separation of Aqueous Ionic Liquids in Covalent Organic Frameworks for Thermal Switchable Proton Conductivity.

Covalent organic frameworks (COFs) have regular channels that can accommodate guest molecules to provide highly conductive solid electrolytes. However, designing smart, conductive COFs remains a great challenge. Herein, we report the first example of PEG-functionalized ionic liquids (ILs) anchored on the COF walls by strong hydrogen bonding to fabricate thermally responsive COFs (ILm@COF). We found that similar to the traditional IL/water mixture, the ILs undergo lower critical solution temperature (LCST)-type phase behavior within COF nanopores under high moisture levels. However, the phase separation temperature of aqueous IL decreases in COF channels due to the strong interaction between the IL and COF. Thus, the proton conductivity of ILm@COF can be reversibly switched by phase miscibility and separation in COF nanopores, and there is no obvious decrease even after 20 switching cycles. Our work provides important clues for understanding liquid-liquid phase separation in a confined nanospace and opens a new pathway to switchable proton conductivity.

Isomerization of DASA Molecules in the Nanopores of Metal-Organic Frameworks: What Determines Its Reversibility?

In recent years, light-responsive molecules have been incorporated in metal-organic frameworks (MOFs) to fabricate light-responsive intelligent devices, where reversible isomerization of the guest molecules in the nanopores is crucial. However, how to design a porous environment of MOFs to achieve a reversible isomerization remains unknown until now. In this work, donor-acceptor Stenhouse adducts (DASAs), a new kind of visible light responsive compound, were confined in the nanopores of different MOFs to study their isomerization upon visible-light irradiation/mild heating. We found that the polarity of the pore environment is the key to control the reversibility of isomerization of such guest molecules. Under the guidance of this principle, MIL-53(Al) was screened to investigate the proton conductivity and switching performance of the DASA-confined MOF. The proton conductance was up to 0.013 S cm-1 at 80 °C and 98 % RH, and at least 30 switching cycles were achieved thanks to the Grotthuss-type mechanism and the low polarity of MIL-53(Al) pore environment.

Distribution and spread of the mobilised RND efflux pump gene cluster tmexCD-toprJ in clinical Gram-negative bacteria: a molecular epidemiological study.

BACKGROUND: TMexCD1-TOprJ1, which is associated with phenotypic resistance to multiple classes of antibiotics, is a transmissible resistance-nodulation-division (RND) family efflux pump. However, the prevalence and genomic and phenotypic characteristics of clinical isolates with this important resistance determinant are poorly understood. In this study, we aimed to survey tmexCD-toprJ among clinical Gram-negative isolates collected from hospitals in China between 1991 and 2020 and characterise tmexCD-toprJ-positive clinical isolates. METHODS: We conducted online data retrieval and active nationwide surveillance in China to screen tmexCD-toprJ-positive strains. We characterised tmexCD-toprJ-positive clinical strains for their antimicrobial susceptibility, genetic and functional characteristics, and the potential inter-species transmission route of tmexCD-toprJ with whole genome sequencing and bioinformatics analyses. The function of tmexCD-toprJ in Pseudomonas sp and Proteus sp was investigated by tmexD gene knockdown using an isopropylthio-ß-galactoside-inducible CRISPR interference system. FINDINGS: Data retrieval obtained 53 strains carrying tmexCD-toprJ, comprising 32 Pseudomonas spp, 11 Klebsiella pneumoniae, one Aeromonas spp, one Citrobacter freundii, and one uncultured bacterium from diverse niches. 48 (0·64%) of 7517 clinical isolates from China, including seven Klebsiella spp, one Proteus mirabilis, and 40 Pseudomonas spp, carried tmexCD-toprJ. These isolates exhibited multidrug resistance phenotypes and co-harboured resistance genes, such as mcr and carbapenemases genes. tmexCD-toprJ was encoded on both plasmids and chromosomes in all Klebsiella spp that carried plasmid-borne tmexCD-toprJ (n=7), P mirabilis carried chromosome-borne tmexCD-toprJ, and Pseudomonas spp carried either plasmid-borne (n=19) or chromosome-borne (n=21) ones. tmexCD-toprJ had undergone clonal and horizontal transmission among clinical pathogens. Eight different types of genetic context of tmexCD-toprJ were identified, each of which was associated with different mobile elements, including IntI, IS6100, TnAs1-like, ISRor5, ISVsa3, ISCfr-like, Tn5393, and IS222-like, which might facilitate its transmission. Knockdown of tmexD led to a four times decrease in tigecycline minimum inhibitory concentrations in both Pseudomonas spp and Proteus spp. INTERPRETATION: Our study provides evidence to suggest that tmexCD-toprJ contributes to the antimicrobial resistance phenotypes in different bacterial species. tmexCD-toprJ has disseminated among diverse species of clinical pathogens, which warrants timely monitoring in clinical pathogens. FUNDING: National Natural Science Foundation of China, Guangdong Major Project of Basic and Applied Basic Research, Natural Science Foundation of Jiangsu Province.

A visible light/heat responsive covalent organic framework for highly efficient and switchable proton conductivity.

In recent years, covalent organic frameworks (COFs) have attracted enormous interest as a new generation of proton-exchange membranes, chemical sensors and electronic devices. However, to design high proton conductivity COFs, especially those with stimulus responsive performance remains a great challenge. Here, the first example of a light/heat switchable COF (COF-HNU9) has been synthesized by grafting a donor-acceptor Stenhouse adduct (DASA) within the channels of a ß-ketoenamine-based COF. DASA groups in the nanopores of COF-HNU9 undergo a reversible open-closed photoisomerization upon visible light irradiation and are recovered by heating. Thus, COF-HNU9 exhibits not only a remarkably high proton conductivity, but also a highly effective switching performance. Under visible light irradiation at 98% RH, the proton conductivity of COF-HNU9 increases by three orders of magnitude at 25 °C and is up to 0.02 S cm-1 at 80 °C. Furthermore, the proton conductivity does not display any significant decrease even after 20 switching cycles. These results have been rationalized by a Grotthuss-type mechanism and verified by DFT calculations. The stimuli-responsive COF is conceptually confirmed by an optical control device with the light/heat switching proton conductive COF-HNU9 film, which is able to remote-control the illumination and switching off of an LED lamp without any current amplifier.

A record ammonia adsorption by calcium chloride confined in covalent organic frameworks.

Ammonia is a vital chemical raw material, but it is also a highly toxic environmenal pollutant. However, its highly efficient uptake and reversible release is a challenge. Herein, we have designed and synthesized a series of hybrid materials for efficient NH3 capture by confining calcium chloride (CaCl2) in a porous covalent organic framework (COF). A high capture capacity of 26.5 mmol g-1 is obtained at 25 °C and 1 bar, which is the highest value among existing porous materials, and NH3 can be easily desorbed at 80 °C under vacuum for 2 h. Particularly, the hybrid COF is highly efficient for the absorption of low NH3 content. Such excellent performance is ascribed to the highly dispersion of CaCl2 in the pores of the COF, and coordinating interaction of NH3 to Ca2+ together with hydrogen bond interaction between NH3 and Cl-.

A reversible conductivity modulation of azobenzene-based ionic liquids in aqueous solutions using UV/vis light.

Photo-induced conductivity modulation of stimuli-responsive materials is of great importance from the viewpoint of fundamental research and technology. In this work, 5 new kinds of azobenzene-based photo-responsive ionic liquids were synthesized and characterized, and UV/vis light modulation of their conductivity was investigated in an aqueous solution. The factors affecting the conductivity modulation of the photo-responsive fluids, such as photo-isomerization efficiency, photo-regulation aggregation, concentration and chemical structure of the ionic liquids, were examined systematically. It was found that the conductivity of the ionic liquids in water exhibited a significant increase upon UV light irradiation and the ionic liquids with a shorter alkyl spacer in the cation showed a more remarkable photo-induced conductivity enhancement with a maximum increase of 150%. In addition, the solution conductivity was restored (or very close) to the initial value upon an alternative irradiation with visible light. Thus, the solution conductivity can be modulated using alternative irradiation with UV and visible light. Although the reversible photo-isomerization of the azobenzene group under UV/vis irradiation is the origin of the conductivity modulation, the photo-regulated aggregation of the ionic liquid in water is indispensable for the maximum degree of conductivity modulation because UV irradiation can weaken, even break the aggregated cis-isomers of the ionic liquids in an aqueous solution.

Visible Light-Controlled Inversion of Pickering Emulsions Stabilized by Functional Silica Microspheres.

A new class of donor-acceptor Stenhouse adduct (DASA)-functionalized silica microspheres (SMs) is designed and described to formulate Pickering emulsions with inversion property and large polarity change upon visible light irradiation. By tuning the hydrophilicity of the functional SM particles with visible light, these Pickering emulsions can easily perform inversion from water-in-oil to oil-in-water. The inversion performance of the emulsions is ascribed to DASA photoisomerization from an extended, hydrophobic, and intensely purple-colored triene to a compact, zwitterionic, and colorless cyclopentenone upon irradiation with visible light. This unique inversion behavior has been applied to control encapsulation and the release of fluorescein sodium salt.