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Epigallocatechin-3-gallate (EGCG) is a vital kind of catechin with high bioactive activities, however, limited research has been conducted to elucidate the molecular basis of EGCG biotransformation by Aspergillus niger and the underlying regulatory mechanisms. In this study, A. niger RAF106, isolated from Pu-erh tea, was applied to conduct the EGCG fermentation process, and the samples were collected at different fermentation times to determine the intermediary metabolites of EGCG and the metabolome as well as physiological activity changes of A. niger RAF106. The results demonstrated that EGCG enhances the growth of A. niger RAF106 by promoting conidial germination and hyphae branching. Meanwhile, metabolomic analyses indicated that EGCG significantly regulates the amino acid metabolism of A. niger RAF106. Furthermore, metabolomic analyses also revealed that the levels of original secondary metabolites in the supernatant of the cultures changed significantly from the fermentation stage M2 to M3, in which the main differentially changed metabolites (DCMs) were flavonoids. Most of these flavonoids exhibited antioxidant properties and thus increased the radical scavenging activity of the supernatant of the cultures. In addition, we also found several intermediary metabolites of EGCG, GA, and EGC, including oolonghomobisflavan A, (-)-Epigallocatechin 3, 5-di-gallate, (-)-Epigallocatechin 3-(3-methyl-gallate) (-)-Catechin 3-O-gallate, 4'-Methyl-(-)-epigallocatechin 3-(4-methyl-gallate), myricetin, prodelphinidin B, 7-galloylcatechin, and 3-hydroxyphenylacetic acid. These findings contribute to improving the bioavailability of EGCG and help mine highly active metabolites, which can be used as raw materials for the development of pharmaceutical intermediates or functional foods. In addition, the results also provide a theoretical basis for better control of the risk of A. niger origin and the regulatory mechanisms of the biotransformation process mediated by A. niger.

In this work, we report that FeCl3, a benign Lewis acid, could be used as a catalyst in the γ-valerolactone (GVL)/H2O solvent system to facilitate efficient pretreatment of Chinese silvergrass. The results showed that approximately 87.8% of xylan and 75.8% of lignin were removed by 80% GVL under 170 °C using 50 mM FeCl3 as a catalyst. The solubilized carbohydrate fractions in the pretreatment liquor were mainly composed of monosaccharides (glucose and xylose) rather than oligosaccharides (glucose oligomers and xylose oligomers) and inhibitors (furfural, hydroxymethylfurfural, formic acid and acetic acid). Moreover, FeCl3 decreased the transesterification of GVL with hydroxyl groups on the cellulose surface and resulted in more easily degraded cellulose. Approximately 96.5% of cellulose and 92.2% of xylan in the pretreated Chinese silvergrass were converted to fermentable monosaccharides by 10 FPU/g cellulases. This study indicated that FeCl3-catalyzed GVL solvent could efficiently deconstruct biomass and increase the cellulose digestibility, which provided a novel, GVL-based pretreatment system for processing biomass.

Lignin is a natural aromatic polymer of

Citrobacter werkmanii, an aerobe and mesophilic Proteobacterium, is universal in industrial putrefaction, coastal water, and human blood. Our previous studies have discovered that outer membrane protein X (OmpX) of C. werkmanii is involved in calcium response, but the underlying mechanisms and its molecular characteristics remain elusive. To that end, the ompX gene was deleted from the genome of C. werkmanii and its phenotypic variations were thoroughly investigated in conjunction with the wild type (WT) and complementary strains using biochemical and molecular techniques such as RNA-Seq, respectively. The results demonstrated that deleting ompX reduces biofilm formation on polystyrene and glass surfaces. Meanwhile, ΔompX's swimming ability but not for its twitching or swarming abilities, was also reduced on semi-solid plates compared with WT, which was caused by inhibition of flagellar assembly genes, such as flgC, flhB, and fliE, etc. Furthermore, ompX inactivation altered susceptibility to various bactericide classes, as well as responses to Ca

Background Bacterial outer membrane proteins (Omps) play crucial roles in environmental senses, stress responses and substance transportation. Our previous study has found that OmpA contributes to planktonic growth, biocide resistance, biofilm formation and swimming motility in Citrobacter werkmanii, whereas molecular functions of OmpF in this strain remain largely unknown. Hence, in this work, the ompF was firstly knocked out from the genome of C. werkmanii using a homologous recombination method and its phenotypical alternations of ∆ompF were then thoroughly characterized with the parental wild type (WT) and complementary (∆ompF-com) strains using biochemical and molecular approaches. Results The results demonstrated that the swimming ability of ∆ompF was reduced on semi-solid plates compared to WT due to down-regulation of flgC, flgH, fliK and fliF. Meanwhile, the deletion of ompF leads to a decrease of biofilm formation on both glass and polystyrene surfaces because of the reduced cell aggregation. Additionally, the inactivation of ompF also induced different osmotic stress (carbon sources and mental ions) responses of its biofilms comparing with that of WT and ∆ompF-com. Finally, a total of 6 maltose metabolic genes of lamB, malE, malK, malG, malM and malF were all up-regulated in ∆ompF. Gene knockout and HPLC results further revealed that MalEFGK2 cluster mainly contributed to maltose transportation in C. werkmanii. What’s more, for the first time, we have found that the upstream promoter of OmpF and its transcription can be combined with and regulated by MalT. Conclusions Overall, OmpF contributes to a variety of biochemical processes and molecular functions in C. werkmanii and even may be acted as a targeted site to restrain biofilm formation.

Mounting evidence has shown that antimicrobial agents can interfere synergistically with bacterial viability and proliferation when acting together at both the planktonic and biofilm levels without clear underlying molecular mechanisms. Here, multiplexed proteomics by iTRAQ was used to study the interplay between two biocides, the isothiazolone 1,2-benzisothiazolin-3-one (BIT) and the chelating agent disodium ethylenediaminetetraacetic acid (EDTA-2Na), employing the Citrobacter werkmanii as a model system. We first confirmed that these two biocides act synergistically on this bacterial species and then extracted the proteomic profiles of C. werkmanii cells in the presence of BIT, EDTA-2Na, and their combinations. In particular, we identified 43 core proteins that are differentially expressed in all three conditions simultaneously. Meanwhile, we found that these core proteins are consistently up-regulated when these two biocides are present, but not for single biocides, where we found a balanced mix of up- and down-regulation. Meanwhile, most of the deletion mutants of the core DEPs exhibited biofilm growth inhibition under joint biocide action, while their response was very heterogenous, with respect to the wild-type strain. Together, our results show that while BIT and EDTA-2Na act on multiple protein targets, they interact synergistically at the protein level in a very consistent manner. SIGNIFICANCE: Our preliminary experiments have demonstrated that a combination of 1,2-benzisothiazolin-3-one (BIT) and EDTA-2Na shows higher inhibitory effects on planktonic growth and biofilm formation in both C. werkmanii and Staphylococcus aureus than when these two biocides act alone. However, the mechanistic basis of such synergistic interaction is still unknown. Therefore, the key proteins involved in the above-mentioned enhanced antimicrobial synergy were elucidated using multiplexed proteomics analysis by isobaric tags for relative and absolute quantification (iTRAQ). Our results reveal that the joint action of BIT and EDTA-2Na induces consistent protein expression alteration in a set of core proteins of C. werkmanii, which underlies a strong synergistic antimicrobial effect, which increase our understanding of the action modes of BIT and EDTA-2Na as well as their combinations.

Diallyl disulfide (DAS) from garlic did not affect the growth dynamics of Pseudomonas aeruginosa PAO1; however, DAS inhibited transcription of most of the quorum sensing (QS) system genes, including lasR, rhlI/rhlR, and pqsABCDE/pqsR; thus, biosynthesis of the signal molecules C4‐HSL (encoded by rhlI) and PQS (encoded by pqsABCDE) was inhibited.DAS suppressed the production of some virulence factors toxic to the host and enhanced the production of some nutrition factors beneficial to the host. These actions of DAS may be due to its thioether group., Summary Diallyl sulfide (DAS) and diallyl disulfide (DADS), two constituents of garlic, can inhibit quorum sensing (QS) systems of Pseudomonas aeruginosa. However, the differences in the mechanism of QS inhibition between DAS and DADS, and the functional chemical groups of these sulfides that contribute in QS inhibition have not been elucidated yet. We assumed that the sulfide group might play a key role in QS inhibition. To prove this hypothesis and to clarify these unsolved problems, in this study, we synthesized diallyl ether (DAE), and compared and investigated the effects of DAS and DAE on the growth and production of virulence factors, including Pseudomonas quinolone signal (PQS), elastase and pyocyanin, of P. aeruginosa PAO1. Transcriptome analysis and qRT‐PCR were used to compare and analyse the differentially expressed genes between the different treatment groups (DAS, DAE and control). The results indicated that DAS did not affect the growth dynamics of P. aeruginosa PAO1; however, DAS inhibited transcription of most of the QS system genes, including lasR, rhlI/rhlR and pqsABCDE/pqsR; thus, biosynthesis of the signal molecules C4‐HSL (encoded by rhlI) and PQS (encoded by pqsABCDE) was inhibited. Furthermore, DAS inhibited the transcription of virulence genes regulated by the QS systems, including rhlABC, lasA, lasB, lecA and phzAB, phzDEFG, phzM and phzS that encode for rhamnolipid, exoprotease, elastase, lectin and pyocyanin biosynthesis respectively. DAS also enhanced the expression of the key genes involved in the biosynthesis of three B vitamins: folate, thiamine and riboflavin. In conclusion, DAS suppressed the production of some virulence factors toxic to the host and enhanced the production of some nutrition factors beneficial to the host. These actions of DAS may be due to its thioether group. These findings would be significant for development of an effective drug to control the virulence and pathogenesis of the opportunistic pathogen P. aeruginosa.

In the title complex molecule, [Cd(C13H11N4O2)2], the Cd atom is coordinated in a distorted octahedral geometry by two tridentate ligands synthesized from 2-hydroxybenzohydrazide and 1-(pyrazin-2-yl)ethan-1-one. The molecule has twofold crystallographic symmetry and is isomorphous to its Mn, Co, Ni, Cu and Zn counterparts.

The development of long-wavelength emission carbon dots as a label-free sensing platform for the direct detection of glutathione (GSH) is still of great challenge. Herein, novel rose-red fluorescence carbon dots (wCDs) with a long-wavelength emission of 654 nm were successfully synthesized by a one-step microwave-solvothermal treatment of biomass Wedelia trilobata. The biomass-based wCDs could sensitively respond to copper ions (Cu2+) with a limit of detection (LOD) of 0.22 μM, which could be utilized as the fluorescence turn off–on sensor for the detection of GSH through recovering their fluorescence quenched by Cu2+. Moreover, the label-free wCDs with abundant active functional groups could be used to construct a direct quenching platform for the sensitive and selective detection of GSH by the dynamic quenching effect within 20 s without further modification, displaying a good linear range of 0–3.0 mM with a LOD of 35 μM that fits well with the GSH content in living cells (0.5–10 mM). Additionally, confocal imaging of living cells indicates that as-prepared wCDs with high photobleaching resistance and favorable biocompatibility possess the capacity for real-time monitoring of the variations of intracellular Cu2+ and GSH levels, and distinguishing cancerous cells from normal cells owing to the overexpression of GSH in cancerous cells. This study opens a new vision for developing an efficient and sensitive fluorescent sensor for the detection of GSH, and demonstrates the great prospects for wCDs in ion/molecule tracking, bioimaging and cancer diagnosis.

Through our previous study, we found an up-regulation in the expression of nitrite reductase (nirS) in the isothiazolone-resistant strain of Pseudomonas aeruginosa. However, the definitive molecular role of nirS in ascribing the resistance remained elusive. In the present study, the nirS gene was deleted from the chromosome of P. aeruginosa ATCC 9027 and the resulting phenotypic changes of ΔnirS were studied alongside the wild-type (WT) strain under aerobic conditions. The results demonstrated a decline in the formations of biofilms but not planktonic growth by ΔnirS as compared to WT, especially in the presence of benzisothiazolinone (BIT). Meanwhile, the deletion of nirS impaired swimming motility of P. aeruginosa under the stress of BIT. To assess the influence of nirS on the transcriptome of P. aeruginosa, RNA-seq experiments comparing the ΔnirS with WT were also performed. A total of 694 genes were found to be differentially expressed in ΔnirS, of which 192 were up-regulated, while 502 were down-regulated. In addition, these differently expressed genes were noted to significantly enrich the carbon metabolism along with glyoxylate and dicarboxylate metabolisms. Meanwhile, results from RT-PCR suggested the contribution of mexEF-oprN to the development of BIT resistance by ΔnirS. Further, c-di-GMP was less in ΔnirS than in WT, as revealed by HPLC. Taken together, our results confirm that nirS of P. aeruginosa ATCC 9027 plays a role in BIT resistance along with biofilm formation and further affects several metabolic patterns under aerobic conditions.

The process and mechanism of heavy metal flocculation with extracellular polymeric substances (EPS) secreted by microorganisms, are crucial to their fate in natural environment, wastewater treatment and soil bioremediation applications. However, the structural features of EPS and the relationship between these features and the flocculation process and mechanism remain unclear. In the present study, structural features of the microbial product poly-γ-glutamic acid (γ-PGA) complexed with the heavy metal ions Pb

Heavy metal pollution is widespread and persistent, and causes serious harm to the environment. Pseudomonas putida, a representative environmental microorganism, has strong resistance to heavy metals due to its multiple efflux systems. Although the functions of many efflux systems have been well-studied, the relationship between them remains unclear. Here, the relationship between the Czc and Cad systems that are predominantly responsible for cadmium efflux in P. putida KT2440 is identified. The results demonstrated that CzcR3, the response regulator of two-component system CzcRS3 in the Czc system, activates the expression of efflux pump genes czcCBA1 and czcCBA2 by directly binding to their promoters, thereby helping the strain resist cadmium stress. CzcR3 can also bind to its own promoter, but it has only a weak regulatory effect. The high-level expression of czcRS3 needs to be induced by Cd2+, and this relies on the regulation of CadR, a key regulator in the Cad system, which showed affinity to czcRS3 promoter. Our study indicates that the Cad system is involved in the regulation of the Czc system, and this relationship is important for maintaining the considerable resistance to cadmium in P. putida.

Numerous attempts have been utilized to unveil the occurrences of antibiotic resistance genes (ARGs) in human-associated and non-human-associated samples. However, spoiled household chemicals, which are usually neglected by the public, may be also a reservoir of ARGs because of the excessive and inappropriate uses of industrial drugs. Based upon the Comprehensive Antibiotic Research Database, a metagenomic sequencing method was utilized to detect and quantify Antibiotic Resistance Ontology (AROs) in six spoiled household chemicals, including hair conditioner, dishwashing detergent, bath shampoo, hand sanitizer, and laundry detergent. Proteobacteria was found to be the dominant phylum in all the samples. Functional annotation of the unigenes obtained against the KEGG pathway, eggNOG and CAZy databases demonstrated a diversity of their functions. Moreover, 186 types of AROs that were members of 72 drug classes were identified. Multidrug resistance genes were the most dominant types, and there were 17 AROs whose resistance mechanisms were categorized into the resistance-nodulation-cell division (RND) antibiotic efflux pump among the top 20 AROs. Moreover, Proteobacteria was the dominant carrier of AROs with the primary resistance mechanism of antibiotic efflux. The maximum temperature of the months of collection significantly affected the distributions of AROs. Additionally, the isolated individual bacterium from spoiled household chemicals and artificial mixed communities of isolated bacteria demonstrated diverse resistant abilities to different biocides. This study demonstrated that there are abundant microorganisms and a broad spectrum profile of AROs in spoiled household chemicals that might induce a severe threat to public healthy securities and merit particular attention.

Staphylococcus aureus is the causative agent of numerous and varied clinical infections. Crude aqueous extracts of Melia azedarach fruits inhibit the planktonic growth and initial biofilm formation of S. aureus in a dose-dependent manner. Moreover, the biofilm topologies became sparse and decreased as the concentration of the aqueous extracts increased. RNA-Seq analyses revealed 532 differentially expressed genes (DEGs) after S. aureus exposure to 0.25 g/ml extracts; 319 of them were upregulated, and 213 were downregulated. The majority of DEGs were categorized into abundant sub-groups in the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Finally, untargeted UHPLC-MS/MS analyses of the aqueous extracts of M. azedarach fruits demonstrated a highly complex profile in positive and negative electrospray ionization modes. The extracts primarily consisted of lipids and lipid-like molecules, organic acids and their derivatives, phenylpropanoids, polyketides, organoheterocyclic compounds, and benzenoids annotated by abundant lipid maps and KEGG pathways. Overall, this study provides evidences that the aqueous extracts of M. azedarach fruits can control S. aureus infections and sought to understand the mode of action of these extracts on S. aureus.

Nitric oxide (NO) reductase (NorCB) of Pseudomonas aeruginosa is an essential enzyme that metabolizes NO and alleviates anaerobic NO toxicity during denitrification processes under anaerobic conditions. However, the molecular functions of norCB in the presence of oxygen are poorly understood. This study utilized norCB knockout from P. aeruginosa ATCC 9027 to analyze the resulting phenotypic changes of ΔnorCB in comparison to the wild-type parental strain (WT) and the complementary strain (ΔnorCB-com). The results demonstrated an increase in planktonic growth and biofilm formation by ΔnorCB compared to WT and ΔnorCB-com in the presence of isothiazolones under aerobic conditions. Deletion of norCB led to increased swimming ability and decreased pyocyanin production. Inactivation of norCB also led to an increase of cellular H2 O2 concentration due to decreased activity of its catalases. In addition, the deletion of norCB also influenced the relative expressions of several other genes, including norD, nirS, hmgA, and hpd. These findings provide preliminary evidence that norCB in P. aeruginosa plays an essential role in bacterial life process under aerobic conditions and improves the application of denitrification in the next step.

Cellulose acetate is one of the oldest synthetic thermoplastic polymers and has various applications including usage as films, fibers, and membrane products. Traditionally, cellulose acetate is pre...

Nanocomposites composed of Ag and γ-Fe2O3 nanoparticles within poly-γ-glutamic acid (γ-PGA) derived amorphous carbon nitride (a-CN) films (Ag + γ-Fe2O3@a-CN) were synthesized by a one-step facile pyrolysis strategy. Transmission electron microscopy analysis shows that Ag and γ-Fe2O3 nanoparticles were obtained in situ and homogeneously dispersed on the a-CN matrix. The average size of nanoparticles was 8.1 nm. The presence of γ-Fe2O3, Ag, and a-CN in the nanocomposite was confirmed by X-ray diffraction analysis, UV–visible absorption spectroscopy, Raman spectroscopy, and X-ray photoelectron spectroscopy. Ag + γ-Fe2O3@a-CN catalyzed the degradation of methyl orange. This catalyst was also recycled ten times by magnetic separation without any loss in efficiency. Ag + γ-Fe2O3@a-CN exhibited strong antibacterial activity against E. coli and S. aureus. It also exhibited excellent antibacterial activity even after 20 times magnetic recycling. This indicates that this is a promising recyclable antibacterial and catalyst for environmental applications.

This study investigated the effects of mold susceptibility, total soluble sugar contents and chemical components of wood fibers on mold resistance of wood-polypropylene composites (WPC) filled with white poplar, moso-bamboo, Chinese fir, Ramin, white pine, red gum, red cedar, and rubberwood. Susceptibilities and resistances to molds were studied by artificially accelerated tests. The total soluble sugar contents and chemical components of wood fibers were analyzed by anthrone colorimetry and gas chromatography-mass spectrometry (GC-MS), respectively. Experimental results showed that the mold resistance of WPC varied significantly with wood fiber species. Higher natural durability of wood fiber resulted in higher mold resistance of WPC, while higher sugar contents decreased the resistance of WPC against molds. Low initial moisture content did not have any influence on the susceptibility of WPC towards fungal attack, but higher final moisture content was the result of heavy mold growth on WPC. GC-MS analysis suggested that different chemical components of wood fiber species affected the mold resistance of WPC either positively or negatively. This study provides an alternative solution to improve the durability of WPC against fungal attack.

Bacterial biofilms, formed on biotic or abiotic surfaces, can lead to serious environmental or medical problems. Therefore, it is necessary to find novel antimicrobial agents to combat biofilms, or more effective combinations of existing biocides. In this study, initial biofilms of Pseudomonas aeruginosa ATCC 9027 and Staphylococcus aureus ATCC 6538 in the presence of xylitol or xylitol and isothiazolones were determined using crystal violet staining in 96-well microplates and confocal laser scanning microscopy. Xylitol and isothiazolones exhibited enhanced synergistic inhibition of initial biofilm formation, and also the structure and production of extracellular polymeric substances by P. aeruginosa ATCC 9027 and S. aureus ATCC 6538 in a dose-dependent manner. In addition, xylitol and isothiazolones inhibited and restored the swimming motility of P. aeruginosa ATCC 9027, respectively. These findings show that a combination of xylitol and isothiazolones exerts pronounced antimicrobial activity against P. aeruginosa and S. aureus biofilms and may be applicable for preventing or reducing bacterial biofilms in vitro.

Chitosan oligosaccharide (COS), a time-dependent antimicrobial carbohydrate, is found antifungal active with a short duration of action due to excessive solubility. We attempted to address this issue by employing a hydrogel as a COS carrier. In this research, macroporous zwitterionic composite cryogels composed of COS and poly(N-methacryl arginine) (PMarg) were fabricated, serving as long-term antifungal dressings. Firstly, Marg was synthesized and characterized by Fourier transform infrared spectroscopy (FT-IR), 1H and 13C nuclear magnetic resonance (NMR), and high-resolution mass spectrometry (HRMS). Then, the COS/PMarg cryogels were prepared by redox initiation cryopolymerization. The macroporous morphology of the cryogels was confirmed by scanning electron microscope (SEM) with pore size varying from 20.86 to 50.87 μm. FTIR indicated that hydrogen bonding formed between COS and PMarg, and the interaction elevated thermal stability of the cryogels as evidenced by thermal-gravimetric analysis (TGA). Swelling capacity, mechanical properties, and COS release behavior of the COS/PMarg cryogels were investigated. With the release of COS, the antifouling activity of the cryogel increased. Antimicrobial tests indicated the COS/PMarg cryogel could effectively inhibit the proliferation of Candida albicans. It demonstrated that the macroporous zwitterionic COS/PMarg composite cryogel might be a potential antifungal dressing with sequential “sterilization-release” capacity.

The structure of a cadmium(II) aroylhydrazone complex, viz. bis­{2-hy­droxy-N′-[1-(pyrazin-2-yl)ethyl­idene]benzohydrazidato}cadmium(II) is described., In the title complex mol­ecule, [Cd(C13H11N4O2)2], the Cd atom is coordinated in a distorted octa­hedral geometry by two tridentate ligands synthesized from 2-hy­droxy­benzohydrazide and 1-(pyrazin-2-yl)ethan-1-one. The mol­ecule has twofold crystallographic symmetry and is isomorphous to its Mn, Co, Ni, Cu and Zn counterparts.

Inspired by antimicrobial peptides (AMP) which could alleviate drug resistance pressure, antimicrobial peptide mimics (AMPMs) were designed timely. Here, carboxymethyl cellulose (CMC) -based AMPMs were constructed by introducing different diamines on CMC effectively. Firstly, CMC was degraded to be oligomers with different molecular weights, followed by amination reactions with different diamines respectively. After protonation, a series of AMPMs with different structures were synthesized successfully. Their antibacterial effect has been evaluated by dynamic growth curves and microdilution method. The images snapped by the confocal laser scanning microscope and transmission electron microscope have fully proved its great lethality. And the antibacterial mechanism measured by flow cytometry analysis and zeta potential detection demonstrated that the destruction of membrane potential leads to bacteria death. The excellent blood compatibility and negligible drug resistance has also been confirmed. In addition, the synthesis method is simple and environmental-friendly.

Absract Intrinsic dual-emission carbon dots (CDs) with long wavelength and self-calibration attributes are critical in fluorescence sensing and imaging, but their synthesis remains a great challenge. Here, nitrogen/sulfur-doped dual-emission CDs (NS-CDs) of different sizes were controllably synthesized by solvothermally treatment of sodium alginate (SA) and glutathione (GSH) in formamide. Interestingly, these as-prepared NS-CDs exhibit enchanting dual-emission behavior at 480 and 650 nm, tunable fluorescence, superior photostability, and high nitrogen level up to 19.57%. The decreasing mass ratio of SA/GSH for the synthesis can lead to larger size and lower graphitic nitrogen for NS-CDs, ultimately giving the enhanced photoluminescence (PL) and tunable fluorescence changing from cyan to white-blue. The selected dual-emission NS-CDs showed superior selectivity and sensitivity for ratiometric Fe3+ sensing with a detection limit as low as 0.56 μM because of the aggregation of NS-CDs induced by Fe3+, and achieved the Fe3+ detection in environment water. Moreover, the fluorescence-tunable NS-CDs possessing plenteous hydrophilic groups exhibit excellent biocompatibility and unprecedented localized capability for specific cell membrane imaging. This study designs a facile and economical strategy to prepare dual-emission CDs with long wavlength, tunable fluorescence and high nitrogen level, which present bright prospects in environmental monitoring and cell imaging.

Ionic liquids (ILs), also known as green solvents, are widely acknowledged in several fields, such as chemical separation, synthesis, and electrochemistry, owing to their excellent physiochemical properties. However, their poor biodegradability may lead to environmental and health risks, posing a severe threat to humans, thus requiring further research. In this study, the biotoxicities of the imidazolium-based ILs were evaluated in Tetrahymena pyriformis. Moreover, IL detoxification was investigated by addition of glutathione (GSH), cysteine, and nicotinamide adenine dinucleotide (NADH). Reactive oxygen species (ROS) initiated by different IL types caused damage to Tetrahymena, while glutathione, cysteine, and NADH eliminated ROS, achieving the detoxification purposes. Detoxification results showed that NADH exhibited the best detoxification ability, followed by glutathione and cysteine. Finally, RT-PCR results suggested that metallothionein might have participated in IL detoxification.

The genus Citrobacter is commonly found in environmental and industrial settings, some members of which have been used for bioremediation of heavy metals owing to the absorption ability of their biofilms. Although our previous studies have found that the outer membrane protein A (OmpA) contributes to the process of Citrobacter werkmanii biofilm formation, the underlying mechanisms remain elusive. Therefore, we deleted ompA from the genome of C. werkmanii and investigated its phenotypes in comparison to the wild type strain (WT) and the complementary strain using biochemical and molecular techniques including RNA-Seq. Our results demonstrated that the deletion of ompA led to an increase in biofilm formation on both polystyrene and glass surfaces due to upregulation of some biofilm formation related genes. Meanwhile, swimming ability, which is mediated by activation of flagellar assembly genes, was increased on semi-solid plates in the ∆ompA strain when compared with WT. Additionally, inactivation of ompA also caused increased 1,2-benzisothiazolin-3-one (BIT) resistance, differential responses to Ca2+ stress, curli protein expression and cellulose production. Finally, ∆ompA caused differential expression of a total of 1470 genes when compared with WT, of which 146 were upregulated and 1324 were downregulated. These genes were classified into different Gene Ontology (GO) and KEGG pathways. In summary, ompA in C. werkmanii contributes to a variety of biological functions and may act as a target site to modulate biofilm formation. KEY POINTS: • ompA is a negative regulator for biofilm formation by C. werkmanii. • ompA inhibits swimming motility of C. werkmanii. • ompA deletion causes different expression profiles in C. werkmanii.

Antibiotic pollution has been a serious global public health concern in recent years, photodynamic inactivation is one of the most promising and innovative methods for antibacterial applications that avoids antibiotic abuse and minimizes risks of antibiotic resistance. However, limited by the weak interaction between the photosensitizers and Gram-negative bacteria, the effect of photodynamic inactivation cannot be fully exerted. Herein, photosensitizer chlorin e6-loaded polyethyleneimine-based micelle was constructed. The synergy of electrostatic and hydrophobic interactions between the nanoparticles and the bacterial surface promoted the anchoring of nanoparticles onto the bacteria, resulting in enhanced photoinactivation activities on Gram-negative bacteria. As expected, an eminent antibacterial effect was also observed on the Gram-positive bacteria Staphylococcus aureus. The cellular uptake results showed that photosensitizer was firmly anchored to the bacterial cell surface of Escherichia coli or Staphylococcus aureus by the introduction of branched polyethylenimine-containing nanoparticles. The light-triggered generation of reactive oxygen species, mainly singlet oxygen, from the membrane-bound nanoparticles caused irreversible damage to the bacterial outer membrane, achieving enhanced bactericidal efficiency than free photosensitizer. The study would provide an efficient and promising antimicrobial alternative to prevent overuse of antibiotics and have enormous potential for human healthcare and the environment remediation.

Three aroylhydrazone ligands ((Z)-N′-([2,2′-bithiophen]-5-ylmethylene)-2-hydroxybenzohydrazide, HL1; (Z)-N′-([2,2′-bithiophen]-5-ylmethylene)-3-hydroxybenzohydrazide, HL2; and (Z)-N′-([2,2′-bithiophen]-5-ylmethylene)-4-hydroxybenzohydrazide, HL3) and their complexes with nickel (Ni(L1)2, 1; Ni(L2)2, 2; Ni(L3)2∙DMF, 3) were synthesized and characterized by ESI-MS, NMR, IR, UV–vis and elemental analysis techniques. The molecular structure of ligand (HL2) and complexes 1–3 was confirmed by single crystal X-ray crystallography. The single crystal X-ray structure of complexes 1–3 showed a distorted square planar geometry around the metal center, and the ligands adopt a bidentate chelating mode. The interaction of calf thymus (ctDNA) with nickel(II) complexes was explored using absorption, emission spectrum, viscosity, and circular dichroism methods. These complexes exhibited moderate affinity for ctDNA through groove binding modes. The most efficient DNA binder was complex 2. The interaction of the complexes with DNA has also been supported by molecular docking study and molecular dynamics simulation. An in vitro cytotoxicity study of the complexes found low activity against human cervical (Hela) and breast (MCF-7) cancer cell lines, with the best results for complex 2, where IC50 values are 86 μM and 92 μM respectively.

The association between natural mold resistance and the wood’s chemical components was studied for nine wood species. The mold resistance of the different wood species was tested by artificially accelerated tests and scanning electron microscopy (SEM). The chemical components were analyzed by gas chromatography-mass spectrometry (GC-MS). The results indicated that the sequence of mold resistance of different wood flours was as follows, from greatest to least resistance: spruce, Mongolian pine and camphor, toon and teak, eucalyptus (E. urophylla and E. grandis × E. urophylla), sweetgum, and castor straw fiber. GC-MS analysis indicated that the total contents of the antifungal compounds present in all wood flour extractives were consistent with the sequence of mold resistance of wood flour. This suggested that the natural durability of wood flour against molds was affected by its chemical components.

Garlic oil can disrupt the quorum sensing (QS) pathways of the opportunistic pathogen Pseudomonas aeruginosa; however, the underlying mechanisms for this effect are unclear. Diallyl disulfide (DADS) is one of the most abundant sulfur-containing compounds in garlic oil. This study investigated the effects of DADS on the growth, virulence factor production (elastase, pyocyanin, biofilm, and swarming motility), and essential gene expression of P. aeruginosa PAO1, particularly as they apply to QS and virulence. DADS at 1.28 mg/mL did not affect P. aeruginosa PAO1 growth, although it decreased elastase and pyocyanin production, biofilm formation, and swarming motility. Each of these phenomena is regulated by the three QS systems of P. aeruginosa PAO1 (las, rhl, and pqs). Real-time q-PCR revealed that DADS down-regulated the transcription levels of several important QS genes (lasI, lasR, rhlI, rhlR, pqsA, and pqsR) in the three systems. Furthermore, the transcription levels of QS-regulated virulence genes were also down-regulated. The lasB gene, encoding LasB elastase, is co-regulated by the las, rhl, and pqs systems, and thus the down-regulation of genes across the three systems further down-regulated lasB. Additionally, phzM (encoding pyocyanin), pslB (responsible for the production of a biofilm matrix polysaccharide), and chiC (encoding chitinase) were positively activated by LasR, and a decrease in lasR transcription further down-regulated the transcription of phzM, pslB, and chiC. Hence, DADS inhibits P. aeruginosa PAO1 virulence factors by inactivating the transcription of key genes across three different QS systems.

Many animals such as snails and Chinese giant salamanders can secrete mucus for moisturizing, eliminating swelling and inflammation, and promoting wound healing; this implies that high viscous bio-functional materials inspired by mucus may show potential for biomedical applications. Herein, we developed a maltose-like injectable nanocomposite via the combination of chitosan polysaccharide and clay rectorite for hemostasis. Through the hemostatic property of rectorite, the viscous nanocomposite decreased the in vitro clotting time by 43%. Moreover, an in vitro porcine skin model confirmed that the viscous nanocomposite can stably adhere on skin and impede blood bleeding successfully. Importantly, the internal structure of the nanocomposites, chitosan intercalating into rectorite interlayers, provided a physical cross-linked network to decrease the release of rectorite from the nanocomposite and its invasion into the blood. Therefore, this viscous injectable nanocomposite provided a concept for a facile and sustainable biomaterial for skin hemostasis.