Active site architecture reveals coordination sphere flexibility and specificity determinants in a group of closely related molybdoenzymes.

MtsZ is a molybdenum-containing methionine sulfoxide reductase that supports virulence in the human respiratory pathogen Haemophilus influenzae (Hi). HiMtsZ belongs to a group of structurally and spectroscopically uncharacterized S-/N-oxide reductases, all of which are found in bacterial pathogens. Here, we have solved the crystal structure of HiMtsZ, which reveals that the HiMtsZ substrate-binding site encompasses a previously unrecognized part that accommodates the methionine sulfoxide side chain via interaction with His182 and Arg166. Charge and amino acid composition of this side chain-binding region vary and, as indicated by electrochemical, kinetic, and docking studies, could explain the diverse substrate specificity seen in closely related enzymes of this type. The HiMtsZ Mo active site has an underlying structural flexibility, where dissociation of the central Ser187 ligand affected catalysis at low pH. Unexpectedly, the two main HiMtsZ electron paramagnetic resonance (EPR) species resembled not only a related dimethyl sulfoxide reductase but also a structurally unrelated nitrate reductase that possesses an Asp-Mo ligand. This suggests that contrary to current views, the geometry of the Mo center and its primary ligands, rather than the specific amino acid environment, is the main determinant of the EPR properties of mononuclear Mo enzymes. The flexibility in the electronic structure of the Mo centers is also apparent in two of three HiMtsZ EPR-active Mo(V) species being catalytically incompetent off-pathway forms that could not be fully oxidized.

Multiple Stimuli-Responsive MXene-Based Hydrogel as Intelligent Drug Delivery Carriers for Deep Chronic Wound Healing.

Chronic wound healing is an important and basic issue in medical and healthcare fields. Recently, stimuli-responsive hydrogel systems have emerged as promising drug delivery carriers for wound management. However, given to the limited therapeutic outcomes, new hydrogel systems for efficient wound treatment are urgently needed. Here, the development of a 2D MXene-based hydrogel system for highly efficient photo- and magnetic-responsive drug delivery oriented to deep chronic wounds repair is presented. The intelligent responsive MXene-based hydrogel drug delivery system is composed of MXene-wrapped magnetic colloids and poly(N-isopropyl acrylamide)-alginate dual-network hydrogels. It is demonstrated that the MXene-based hydrogel system exhibits multiple response capability and controllable drug delivery ability, which can reduce the toxic side effects of drugs and promote the wound healing process as well. Notably, the practical performance of the MXene-based hydrogel drug delivery system is demonstrated by applying it to the treatment of the full-thickness cutaneous wound and subcutaneous infected wound of the rat model, which indicates the great prospect in clinical wound healing and other related biomedical fields.

Super-Elastic Magnetic Structural Color Hydrogels.

Structural color hydrogels are promising candidates as scaffold materials for tissue engineering and for matrix cell culture and manipulation, while their super-elastic features are still lacking due to the irreconcilable interfere of the precursor and the self-assembly unit. This hinders many of their practical biomedical applications where elasticity is required. Herein, hydrophilic and size-controllable Fe3 O4 @poly(4-styrenesulfonic acid-co-maleic acid) (PSSMA)@SiO2 magnetic response photonic crystals are fabricated as the assembly units of the structural color hydrogels by orderly packing of core-shell colloidal nanocrystal clusters via a two-step facile synthesis approach. These units are capable of responding instantaneously to an external magnetic field with resistance to interference of ions, thus, by integrating super-elastic hydrogels, super-elastic magnetic structural color hydrogels can be achieved. The structural color arises from the dynamic ordering of the magnetic nanoparticles through the contactless control of external magnetic field, allowing regional polymerization of hydrogels via changing orientation and strength of external magnetic field. These regionally polymerized super-elastic magnetic structural color hydrogels can work as anti-counterfeiting labels with super-elastic identification, which may be widely used in the future.

Radiofrequency heating for powder pasteurization of barley grass: antioxidant substances, sensory quality, microbial load and energy consumption.

BACKGROUND: Young barley grass powder contains abundant nutrition and its antioxidant substances are severely impaired by radiation (60 Co) sterilization. To overcome product quality degradation, radiofrequency pasteurization was conducted using pilot-scale radiofrequency equipment (27 MHz, 6 kW) with electrode gaps of 12, 14 and 16 cm, while hot-air (80 °C) pasteurization was used for comparison. RESULTS: Assessment suggested that uneven radiofrequency heating was improved for the 14 cm electrode gap. With an increase of electrode gap, microbial inactivation needs more energy consumption. A minimum energy consumption of 970 J g-1 was required for 1 log-reduction of colonies. Radiofrequency pasteurization retained better antioxidant substances, lightness (L*), green color (a*) and odors in barley grass powder, compared with hot-air sterilization. Contents of flavonoid and chlorophyll were 5.82 and 4.87 g kg-1 respectively, using the 14 cm electrode gap. Additionally, radiofrequency pasteurization led to an improvement in sourness, bitterness and umami tastes. CONCLUSIONS: Radiofrequency pasteurization would be a superior alternative for the pasteurization of barley grass powder. © 2019 Society of Chemical Industry.

Effect of microwave freeze drying on quality and energy supply in drying of barley grass.

BACKGROUND: Young barley grass leaves are well-known for containing the antioxidant substances flavonoid and chlorophyll. However, low product quality and energy efficiency exist with respect to the dehydration of barley grass leaves. To improve energy supply and the quality of barley grass, microwave heating instead of contact heat was applied for the freeze drying of barley grass at a pilot scale at 1, 1.5 and 2 W g-1 , respectively; After drying, energy supply and quality parameters of color, moisture content, chlorophyll, flavonoids, odors of dried barley grass were determined to evaluate the feasibility of the study. RESULTS: Microwave freeze drying (MFD) allowed a low energy supply and high contents of chlorophyll and flavonoids. A lightness value of 60.0, a green value of -11.5 and an energy supply of 0.61 kW h-1 g-1 were observed in 1.5 W g-1 MFD; whereas drying time (7 h) decreased by 42% compared to contact heating. Maximum content of flavonoid and chlorophyll was 11.7 and 12.8 g kg-1 barley grass. Microwave heating leads to an odor change larger than that for contact heating observed for the freeze drying of barley grass. CONCLUSION: MFD retains chlorophyll and flavonoids, as well as colors and odors of samples, and also decreases energy consumption in the freeze drying of barley grass. © 2017 Society of Chemical Industry.

Carbon Inverse Opal Rods for Nonenzymatic Cholesterol Detection.

Carbon inverse opal rods made from silica photonic crystal rods are used for nonenzymatic cholesterol sensing. The characteristic reflection peak originating from the physical periodic structure works as sensing signals for quantitatively estimating cholesterol concentrations. Carbon inverse opal rods work both in cholesterol standard solutions and human serum. They are suitable for practical use in clinical diagnose.

In vitro investigation of the binding characteristics of dacomitinib to human α 1-acid glycoprotein: Multispectral and computational modeling.

Dacomitinib is a highly selective second-generation tyrosine kinase inhibitor that can irreversibly bind to tyrosine kinase and is mainly used in the treatment of lung cancer. The binding characteristics of dacomitinib with human α 1-acid glycoprotein (HAG) were analyzed by multispectral and computational simulation techniques. The fluorescence spectra showed that dacomitinib can quench the fluorescence of HAG by forming the HAG-dacomitinib complex with a molar ratio of 1:1 (static quenching). At the temperature similar to that of the human body, the affinity of dacomitinib to HAG (8.95 × 106 M-1) was much greater than that to BSA (3.39 × 104 M-1), indicating that dacomitinib will give priority to binding onto HAG. Thermodynamics parameters analysis and driving force competition experiments showed that hydrogen bonding and hydrophobic forces were the major sources for keeping the complex of HAG-dacomitinib stable. The experimental outcomes also showed that the binding of dacomitinib can lead to the loosening of the skeleton structure of HAG, which led to a slight change in the secondary structure, and also reduces the hydrophobicity of the microenvironment of Trp and Tyr residues. The binding sites of dacomitinib on HAG and the contribution of key amino acid residues to the binding reaction were determined by molecular docking and molecular dynamics (MD) simulation. In addition, it was found that there was a synergistic effect between dacomitinib and Mg2+ and Co2+ ions. Mg2+ and Co2+ could increase the Kb of dacomitinib to HAG and prolong the half-life of dacomitinib.

Amine response smartphone-based portable and intelligent polyvinyl alcohol films for real-time detection of shrimp freshness.

Food freshness monitoring is an important component in ensuring food safety for consumers and the food industry. Therefore, there is an urgent need for a portable, low-cost, and efficient detection method to determine the freshness. In this study, polyvinyl alcohol (PVA) was used as polymer carrier to prepare electrospinning film containing curcumin (Cur) and gardenia blue (GB) as intelligent indicator label on food packaging for real-time nondestructive detection of freshness of shrimp. The detection limit of ammonia response is less than or equal to 20 ppm, and the detection time is about 1 min, indicating that it has a sensitive response effect. At the same time, a smartphone application that can identify amines in response to color changes has been developed, and consumers can understand freshness by scanning the label. This study demonstrates the huge potential of smart indicator labels for food freshness monitoring.

The differentiation of new human CD303+ Plasmacytoid dendritic cell subpopulations expressing CD205 and/or CD103 regulated by Non-Small-Cell lung cancer cells.

CD303+ plasmacytoid dendritic cells (pDCs) play an important role in the induction of immune tolerance and antitumor immunity. Here, we focused on the effect of NSCLC cells on the development of CD303+ pDC subsets expressing CD205 and/or CD103. The NSCLC cell line H1299 and primary NSCLC cells were incubated with DCs. The protein expression of costimulatory molecules on CD303+ pDCs, the production of pro-inflammatory and anti-inflammatory cytokines by CD303+ pDCs and the development of CD303+ pDC subsets were detected by using flow cytometry. Coculture with NSCLC cells modulates the protein expression of CD86 and HLA-DR on CD303+ pDCs. Moreover, NSCLC cells suppressed the production of IL-12 and IL-23 but facilitated the secretion of IL-27 and TGF-ß by CD303+ pDCs. There were new CD303+ pDC subsets expressing CD205 and/or CD103 in healthy donors and NSCLC patients: CD303+CD205+CD103+, CD303+CD205+CD103-, CD303+CD205-CD103+ and CD303+CD205-CD103- pDCs. NSCLC cells modulated the differentiation of CD303+ pDC subpopulations by regulating the protein expression of CD205 and/or CD103 on CD303+ pDCs. NSCLC cells may regulate the immune functions of CD303+ pDCs by modulating the expression of costimulatory molecules on DCs and the production of pro-inflammatory/anti-inflammatory cytokines by DCs. NSCLC cells also regulate the development of CD303+ pDC subsets expressing CD205 and/or CD103. These outcomes may reveal a new cellular mechanism leading to the NSCLC-induced immune-suppressive microenvironment.

The Alternative Sigma Factor RpoE2 Is Involved in the Stress Response to Hypochlorite and in vivo Survival of Haemophilus influenzae.

Extracytoplasmic function (ECF) sigma factors underpin the ability of bacteria to adapt to changing environmental conditions, a process that is particularly relevant in human pathogens that inhabit niches where human immune cells contribute to high levels of extracellular stress. Here, we have characterized the previously unstudied RpoE2 ECF sigma factor from the human respiratory pathogen H. influenzae (Hi) and its role in hypochlorite-induced stress. Exposure of H. influenzae to oxidative stress (HOCl, H2O2) increased rpoE2 gene expression, and the activity of RpoE2 was controlled by a cytoplasmic 67-aa anti-sigma factor, HrsE. RpoE2 regulated the expression of the periplasmic MsrAB peptide methionine sulfoxide reductase that, in H. influenzae, is required for HOCl resistance, thus linking RpoE2 to HOCl stress. Interestingly, a HiΔrpoE2 strain had wild-type levels of resistance to oxidative stress in vitro, but HiΔrpoE2 survival was reduced 26-fold in a mouse model of lung infection, demonstrating the relevance of this sigma factor for H. influenzae pathogenesis. The HiRpoE2 system has some similarity to the ECF sigma factors described in Streptomyces and Neisseria sp. that also control the expression of msr genes. However, HiRpoE2 regulation extended to genes encoding other periplasmic damage repair proteins, an operon containing a DoxX-like protein, and also included selected OxyR-controlled genes. Based on our results, we propose that the highly conserved HiRpoE2 sigma factor is a key regulator of H. influenzae responses to oxidative damage in the cell envelope region that controls a variety of target genes required for survival in the host.

Molybdenum Enzymes and How They Support Virulence in Pathogenic Bacteria.

Mononuclear molybdoenzymes are highly versatile catalysts that occur in organisms in all domains of life, where they mediate essential cellular functions such as energy generation and detoxification reactions. Molybdoenzymes are particularly abundant in bacteria, where over 50 distinct types of enzymes have been identified to date. In bacterial pathogens, all aspects of molybdoenzyme biology such as molybdate uptake, cofactor biosynthesis, and function of the enzymes themselves, have been shown to affect fitness in the host as well as virulence. Although current studies are mostly focused on a few key pathogens such as Escherichia coli, Salmonella enterica, Campylobacter jejuni, and Mycobacterium tuberculosis, some common themes for the function and adaptation of the molybdoenzymes to pathogen environmental niches are emerging. Firstly, for many of these enzymes, their role is in supporting bacterial energy generation; and the corresponding pathogen fitness and virulence defects appear to arise from a suboptimally poised metabolic network. Secondly, all substrates converted by virulence-relevant bacterial Mo enzymes belong to classes known to be generated in the host either during inflammation or as part of the host signaling network, with some enzyme groups showing adaptation to the increased conversion of such substrates. Lastly, a specific adaptation to bacterial in-host survival is an emerging link between the regulation of molybdoenzyme expression in bacterial pathogens and the presence of immune system-generated reactive oxygen species. The prevalence of molybdoenzymes in key bacterial pathogens including ESKAPE pathogens, paired with the mounting evidence of their central roles in bacterial fitness during infection, suggest that they could be important future drug targets.

Self-Healable Magnetic Structural Color Hydrogels.

Biologically inspired structural color hydrogels with magnetic- and photothermal-controlled self-healable abilities were fabricated by integrating magnetic-responsive photonic crystals into gelatin hydrogels. The self-healable ability of the hydrogel systems was derived from the magnetic response and light-absorbing abilities of the magnetic nanoparticles. When the hydrogels deteriorate or get damaged, magnetic nanoparticles could absorb heat under near-infrared irradiation and external magnetic fields, which stimulates phase transformation in the hydrogels to fill or heal the hydrogels. In addition, the hydrogel systems were demonstrated with high biocompatibility and plasticity. These features of magnetic self-healable structural color hydrogels make them have broad application prospects in the fields of biological engineering and cell engineering.

Guided cellular orientation concurrently with cell density gradient on butterfly wings.

A simple method to create guided cellular orientation is illustrated by assembling fibroblasts on the dorsal side of M. menelaus wings. Moreover, by inserting the wing into tendon fibroblasts suspension at a tilt angle, guided cellular orientation concurrently with the cell density gradient is formed on the butterfly wings.

Self-assembled colloidal arrays for structural color.

Structural color materials that are colloidally assembled as inspired by nature are attracting increased interest in a wide range of research fields. The assembly of colloidal particles provides a facile and cost-effective strategy for fabricating three-dimensional structural color materials. In this review, the generation mechanisms of structural colors from colloidally assembled photonic crystalline structures (PCSs) and photonic amorphous structures (PASs) are first presented, followed by the state-of-the-art and detailed technologies for their fabrication. The variable optical properties of PASs and PCSs are then discussed, focusing on their spatial long- and short-order structures and surface topography, followed by a detailed description of the modulation of structural color by refractive index and lattice distance. Finally, the current applications of structural color materials colloidally assembled in various fields including biomaterials, microfluidic chips, sensors, displays, and anticounterfeiting are reviewed, together with future applications and tasks to be accomplished.

Effects of ultrasonic pretreatments on quality, energy consumption and sterilization of barley grass in freeze drying.

Barley grass is a plant resource for rehabilitation therapy. Its processing requires retaining nutrition well for rehabilitation cure of consumers. To meet the aim as well as low energy consumption and microbiological safety of products, ultrasonic treatments (UT) were applied to bathing materials at different power levels (10, 30, 45, 60W/L) for 10mins. After treatments, the bathed barley grass (100g) was freeze-dried under vacuum -0.09MPa with fixed power of 2W/g. Parameters of color, microbial colony, energy consumption, glass transition temperature, moisture content, water activity, taste substances, contents of flavonoid and chlorophyll were determined after drying. In contrast with no treatment case, UT (45W/L) decreased drying time by 14% and decreased energy consumption by 19%; UT (60W/L) decreased total microbial colonies by 33%. Also, UT (30W/L) yielded contents of flavonoid (9.2/kg) and chlorophyll (10.5g/kg) of dried sample; UT power (10W/L) yielded the highest L∗(51.5) and the lowest a∗(-9.3) value. Simultaneously, UT leads to a higher glass transition temperature (Tg), lower water activity and produces less sourness and bitterness of dried products. Ultra-sonication is an alternative to improve quality, flavor and energy consumption of barley grass in freeze drying.

Single-Atomic Ruthenium Catalytic Sites on Nitrogen-Doped Graphene for Oxygen Reduction Reaction in Acidic Medium.

The cathodic oxygen reduction reaction (ORR) is essential in the electrochemical energy conversion of fuel cells. Here, through the NH3 atmosphere annealing of a graphene oxide (GO) precursor containing trace amounts of Ru, we have synthesized atomically dispersed Ru on nitrogen-doped graphene that performs as an electrocatalyst for the ORR in acidic medium. The Ru/nitrogen-doped GO catalyst exhibits excellent four-electron ORR activity, offering onset and half-wave potentials of 0.89 and 0.75 V, respectively, vs a reversible hydrogen electrode (RHE) in 0.1 M HClO4, together with better durability and tolerance toward methanol and carbon monoxide poisoning than seen in commercial Pt/C catalysts. X-ray adsorption fine structure analysis and aberration-corrected high-angle annular dark-field scanning transmission electron microscopy are performed and indicate that the chemical structure of Ru is predominantly composed of isolated Ru atoms coordinated with nitrogen atoms on the graphene substrate. Furthermore, a density function theory study of the ORR mechanism suggests that a Ru-oxo-N4 structure appears to be responsible for the ORR catalytic activity in the acidic medium. These findings provide a route for the design of efficient ORR single-atom catalysts.

High-Performance Pseudocapacitive Microsupercapacitors from Laser-Induced Graphene.

All-solid-state, flexible, symmetric, and asymmetric microsupercapacitors are fabricated by a simple method in a scalable fashion from laser-induced graphene on commercial polyimide films, followed by electrodeposition of pseudocapacitive materials on the interdigitated in-plane architectures. These microsupercapacitors demonstrate comparable energy density to commercial lithium thin-film batteries, yet exhibit more than two orders of magnitude higher power density with good mechanical flexibility.

Tin Disulfide Nanoplates on Graphene Nanoribbons for Full Lithium Ion Batteries.

A nanocomposite material made of layered tin disulfide (SnS2) nanoplates vertically grown on reduced graphene oxide nanoribbons (rGONRs) has been successfully developed as an anode in lithium ion batteries by a facile method. At a rate of 0.4 A/g, the material exhibits a high discharge capacity of 823 mAh/g even after 800 cycles. It shows excellent rate stability when the current density varies from 0.1 to 3.0 A/g with a Coulombic efficiency larger than 99%. In order to demonstrate the anode material for practical applications, SnS2-rGONR/LiCoO2 full cells were constructed. To the best of our knowledge, this is the first time that a full cell has been successfully developed using metal chalcogenides as an anode. The full cell delivers a high capacity of 642 mAh/g at 0.2 A/g, superior rate, and cycling stability after long-term cycling. Moreover, the full cell has a high output working voltage of 3.4 V. These excellent lithium storage performances in half and full cells can be mainly attributed to the synergistic effect between the highly conductive network of rGONRs and the high lithium-ion storage capability of layered SnS2 nanoplates.

Cobalt nanoparticles embedded in nitrogen-doped carbon for the hydrogen evolution reaction.

There is great interest in renewable and sustainable energy research to develop low-cost, highly efficient, and stable electrocatalysts as alternatives to replace Pt-based catalysts for the hydrogen evolution reaction (HER). Though nanoparticles encapsulated in carbon shells have been widely used to improve the electrode performances in energy storage devices (e.g., lithium ion batteries), they have attracted less attention in energy-related electrocatalysis. Here we report the synthesis of nitrogen-enriched core-shell structured cobalt-carbon nanoparticles dispersed on graphene sheets and we investigate their HER performances in both acidic and basic media. These catalysts exhibit excellent durability and HER activities with onset overpotentials as low as ∼70 mV in both acidic (0.5 M H2SO4) and alkaline (0.1 M NaOH) electrolytes, and the overpotentials needed to deliver 10 mA cm(-2) are determined to be 265 mV in acid and 337 mV in base, further demonstrating their potential to replace Pt-based catalysts. Control experiments reveal that the active sites for HER might come from the synergistic effects between the cobalt nanoparticles and nitrogen-doped carbon.