Layered Double Hydroxide-Based PdCux@LDH Alloy Nanozyme for a Singlet Oxygen-Boosted Sonodynamic Therapy.

Redox nanozymes have demonstrated tremendous promise in disrupting cellular homeostasis toward cancer therapy, but a dysfunctional competition of diverse activities makes it normally restricted by the complex tumor microenvironment (TME). As palladium nanocrystals can achieve the precise regulation of the enzyme-like activity by regulating exposed crystal planes, noble metal nanoalloys can enhance the enzyme-like activity by promoting electron transfer and enhanced active sites. Herein, bimetallic nanoalloys with optimized enzymatic activity were intelligently designed via the interaction between the Pd and layered double hydroxide, denoted as PdCux@LDH. This PdCux@LDH is able to produce long-lived singlet oxygen (1O2) with high efficiency and selectivity for ultrasound-improved cancer therapy. In addition, this PdCux@LDH nanozyme demonstrated unique surface-dependent multienzyme-mimicking activities for catalyzing cascade reactions: oxidase (OXD)- and catalase (CAT)-mimicking activities. Interestingly, ultrasound (US) stimulation can further improve the dual-enzyme-mimicking activities and impart superior reactive oxygen species (ROS) generation activity, thereby further consuming nicotinamide adenine dinucleotide (NADH) to cause mitochondrial dysfunction, resulting in a highly efficient alloy nanozyme-mediated cancer therapy. This work opens a new research avenue to apply nanozymes for effective sonodynamic therapies (SDT).

Sequence- and Structure-Based Mining of Thermostable D-Allulose 3-Epimerase and Computer-Guided Protein Engineering To Improve Enzyme Activity.

D-Allulose, a functional sweetener, can be synthesized from fructose using D-allulose 3-epimerase (DAEase). Nevertheless, a majority of the reported DAEases have inadequate stability under harsh industrial reaction conditions, which greatly limits their practical applications. In this study, big data mining combined with a computer-guided free energy calculation strategy was employed to discover a novel DAEase with excellent thermostability. Consensus sequence analysis of flexible regions and comparison of binding energies after substrate docking were performed using phylogeny-guided big data analyses. TtDAE from Thermogutta terrifontis was the most thermostable among 358 candidate enzymes, with a half-life of 32 h at 70 °C. Subsequently, structure-guided virtual screening and a customized strategy based on a combinatorial active-site saturation test/iterative saturation mutagenesis were utilized to engineer TtDAE. Finally, the catalytic activity of the M4 variant (P105A/L14C/T63G/I65A) was increased by 5.12-fold. Steered molecular dynamics simulations indicated that M4 had an enlarged substrate-binding pocket, which enhanced the fit between the enzyme and the substrate. The approach presented here, combining DAEases mining with further rational modification, provides guidance for obtaining promising catalysts for industrial-scale production.

Graphene-based photocatalysts for degradation of organic pollution.

Compared with the traditional wastewater treatment technology, semiconductor photocatalysis is a rapidly emerging environment-friendly and efficient Advanced Oxidation Process for degradation of refractory organic contaminants. Single-component semiconductor photocatalysts exhibit poor photocatalytic performance and cannot meet the requirements of wastewater treatment. The combination of semiconductor photocatalysts and Graphene can effectively improve the photocatalytic activity and stability of semiconductor photocatalysts. This review focuses on the synergistic effect of several types of semiconductors with Graphene for photocatalytic degradation of organic pollutants. After a brief introduction of the photodegradation mechanism of semiconductor materials and the basic description of Graphene, the synthesis, characterization and degradation performance of various Graphene-based semiconductor photocatalysts are emphatically introduced.

State-of-the-art strategies and research advances for the biosynthesis of D-amino acids.

D-amino acids (D-AAs) are the enantiomeric counterparts of L-amino acids (L-AAs) and important functional factors with a wide variety of physiological activities and applications in the food manufacture industry. Some D-AAs, such as D-Ala, D-Leu, and D-Phe, have been favored by consumers as sweeteners and fragrances because of their unique flavor. The biosynthesis of D-AAs has attracted much attention in recent years due to their unique advantages. In this review, we comprehensively analyze the structure-function relationships, biosynthesis pathways, multi-enzyme cascade and whole-cell catalysis for the production of D-AAs. The state-of-the-art strategies, including immobilization, protein engineering, and high-throughput screening, are summarized. Future challenges and perspectives of strategies-driven by bioinformatics technologies and smart computing technologies, as well as enzyme immobilization, are also discussed. These new approaches will promote the commercial production and application of D-AAs in the food industry by optimizing the key enzymes for industrial biocatalysts.

Temperature and pH endurable self-assembled camellia-like nanostructure achieved on zinc sheet with superamphiphobicity for fog harvesting.

Easy-implement and low-cost fabrication of super-hydrophobic/super-oleophobic materials is of great significance for efficient fog harvesting. Herein, we propose a simple two-step procedure based on Cu/1-octadecanethiol (ODT) self-assembled monolayer (SAM) modified zinc plates. Interestingly, the whole process mimics the blooming process of flowers in nature: the deposition of copper particle and the subsequent formation of SAM drives the surface undergo gradually stretches and finally blooms to camellia-like nanostructures. The water contact angle (WCA) and oil contact angle (OCA) reach 160 ± 1° and 159 ± 1° respectively, as a result of the formation of layered petal structure that traps air effectively, which is attributed as one of the most important factors for the superamphiphobic effect. In addition to much enhanced facility in fog collection, the materials maintain excellent performance in acid/base environments (1 ≤ pH ≤ 14), broad temperature conditions ranging from -18 °C to 240 °C, and the artificial sea environment, and exhibit capable wear resistance as well as self-cleaning property. The cooperation of all these multiple properties ensures the robustness and stability for efficient fog collection.

Substantial Improvement of an Epimerase for the Synthesis of D-Allulose by Biosensor-Based High-Throughput Microdroplet Screening.

Biosynthesis of D-allulose has been achieved using ketose 3-epimerases (KEases), but its application is limited by poor catalytic performance. In this study, we redesigned a genetically encoded biosensor based on a D-allulose-responsive transcriptional regulator for real-time monitoring of D-allulose. An ultrahigh-throughput droplet-based microfluidic screening platform was further constructed by coupling with this D-allulose-detecting biosensor for the directed evolution of the KEases. Structural analysis of Sinorhizobium fredii D-allulose 3-epimerase (SfDAE) revealed that a highly flexible helix/loop region exposes or occludes the catalytic center as an essential lid conformation regulating substrate recognition. We reprogrammed SfDAE using structure-guided rational design and directed evolution, in which a mutant M3-2 was identified with 17-fold enhanced catalytic efficiency. Our research offers a paradigm for the design and optimization of a biosensor-based microdroplet screening platform.

Directional immobilization of D-allulose 3-epimerase using SpyTag/SpyCatcher strategy as a robust biocatalyst for synthesizing D-allulose.

D-Allulose, as low-calorie rare sugar, possessed several notable biological activities and was biosynthesized by D-allulose 3-epimerase (DAEase). Here, CcDAE from Clostridium cellulolyticum was successfully immobilization via covalent attachment (RI-CcDAE), and Resin-SpyCatcher/SpyTag-CcDAE modular (DI-CcDAE). Both immobilized CcDAEs exhibited higher thermal and pH stabilities than the free form, and they maintained 80.0 % of relative activity after 7 consecutive cycles and 25 days of storage. Predominantly, DI-CcDAE represented superior catalytic efficiency with a 2.4-fold increase of kcat/Km, compared with RI-CcDAE (0.75 s-1 mM-1 vs 0.31 s-1 mM-1). The RI-CcDAE and DI-CcDAE were then applied in mixed fruit Jiaosu to convert D-fructose into D-allulose, which exhibited the productivity of D-allulose 1.08 g/Lh-1 and 1.57 g/Lh-1, respectively. This research provided a promising directional immobilization strategy for DAEase, and robust biocatalyst for production of functional foodstuff containing D-allulose.

Engineering of Acid-Resistant d-Allulose 3-Epimerase for Functional Juice Production.

d-Allulose, a rare sugar and functional sweetener, can be biosynthesized by d-allulose 3-isomerase (DAE). However, most of the reported DAEs exhibit poor resistance under acidic conditions, which severely limited their application. Here, surface charge engineering and random mutagenesis were used to construct a mutant library of CcDAE from Clostridium cellulolyticum H10, combined with high-throughput screening to identify mutants with high activity and resistance under acidic conditions. The mutant M3 (I114R/K123E/H209R) exhibited high activity (3.36-fold of wild-type) and acid resistance (10.6-fold of wild-type) at pH 4.5. The structure-function relationship was further analyzed by molecular dynamics (MD) simulations, which indicated that M3 had a higher number of hydrogen bonds and negative surface charges than the wild type. A multienzyme cascade system including M3 was used to convert high-calorie sugars in acidic juices, and functional juices containing 7.8-15.4 g/L d-allulose were obtained. Our study broadens the manufacture of functional foods containing d-allulose.

Analysis of phosphorus and sulfur effect on soil selenium bioavailability based on diffusive gradients in thin films technique and sequential extraction.

Human intake of selenium (Se) mainly occurs through the food chain, and is largely dependent on the bioavailability of soil Se. Sulfur (S) and phosphorus (P) also as essential nutrients for plants, their antagonistic with Se effects on Se bioavailability should be considered. We conducted pot experiments to investigate the interaction effect on the bioavailability of Se in the soil using a sequential extraction method and diffusive gradients in thin films (DGT). The results showed that the root and shoot Se of pak choi increased at most 340%-360% with S and P application, while the Se uptake by pak choi was slightly inhibited when S and P application was 100 mg kg-1. With high S and P application, pak choi Se had a high bioaccumulation factor (BAF) and low translocation factor (TF), and soil Soluble-Se (SOL-Se) increased 178%-299%, which due to the competitive adsorption of S, P with Se and changes in soil pH that lead to the transformation of soil Se fractions. In addition, the available Se concentration in soil measured by the DGT (CDGT-Se) increased by 866% with exogenous S and P application, and its source was HA-Se. However, CDGT-Se failed to show a good linear relationship with the Se content of pak choi. The application of DGT to assess the bioavailability of Se in soils where Se is present in the steady state needs to be further explored. We discuss the effect of S and P application on the bioavailability of soil Se and provide evidence for agricultural production and rational fertilizer use on Se-rich land.

The auto-oxidative relithiation of spent cathode materials at low temperature environment for efficient and sustainable regeneration.

The reasonable recycling of spent lithium ions batteries is urgently required and beneficial to new energy industry development to approach the "carbon neutral" target. It is urgent to understanding the structural evolution of spent lack lithium cathode materials during direct regeneration technology with low temperatures condition to avoid deficiencies of complex operation in existing technology. Herein, a novel approach was developed for direct regeneration of spent LiCoO2 materials with a successful structural repair and electrochemical performance recovery, which are composed of auto-oxidative process followed by pre-treatment process of dismantling, soaking and sintering. The auto-oxidative system was composed of LiBr as lithium source and dimethyl sulfoxide as solvent and oxygen donor. The recycled LiCoO2 material shows significantly close to capacity retention of 90.79% than that of the commercial LiCoO2 material. Based on the structural evolution mechanism analysis, the novel approach is still expected to be applied into regeneration of other spent cathode materials and guide an efficient and sustainable direction for the recycling of spent lithium ions batteries.

Multi-channel cell co-culture for drug development based on glass microfluidic chip-mass spectrometry coupled platform.

RATIONALE: Cell-based drug assay plays an essential role in drug development. By coupling a microfluidic chip with mass spectrometry (MS), we developed a multifunctional platform. Cell co-culture, cell apoptosis assay, fluorescence and MS detection of intracellular drug absorption could be simultaneously conducted on this platform. METHODS: Three micro-channels were fabricated through photolithography technology to conduct the cell co-culture. Cell apoptosis after drug treatment was assayed by fluorescent probes (Hoechst 33342). Intracellular Dox absorption was analyzed by confocal fluorescent microscopy. With a high voltage (~ 4.5 kV) applied onto the microfluidic chip, the ionization spray was successfully generated by dropping isopropanol onto it. By coupling with a Shimadzu LCMS-2010 A mass spectrometer, intracellular CPA absorption was detected on the microfluidic chip. RESULTS: The microfluidic chip-MS coupled platform showed high biocompatibility. Distinction of cell apoptosis between co-cultured and mono-cultured cells was detected. The results of intracellular drug absorption well explained the different cell apoptosis rate. CONCLUSIONS: Cell-based drug assay was facilely and successfully conducted on the microfluidic chip-MS coupled platform. This technology we have devised could promote MS application in the field of drug development. Copyright © 2016 John Wiley & Sons, Ltd.

Gender differences in cognitive ability associated with genetic variants of NLGN4.

Neuroligin-4 (NL4), encoded by the NLGN4 gene on the X chromosome, is a neuronal-specific brain membrane protein which plays an important role in the formation of functional presynaptic elements and axon specialization. The genetic variants of NLGN4 affect the biological function of NL4, resulting in the manifestation of different psychiatric disorders. The present study investigates the influence of these genetic variants on cognitive performance. The cognitive abilities of 351 subjects were evaluated using the Chinese Wechsler Intelligence Scale Children. The haplotypes were assigned with the PHASE program. The ANOVA method was applied to investigate the relationship between single SNP, the identified target haplotypes and cognitive performance in a random sample. We observed that the X(C) allele of rs5916271 and X(A) allele of the re6638575 carriers had significantly higher cognitive ability performances than the noncarrier boys (p < 0.05). The target haplotype composed of 2 allele (X(CA+)) carriers also displayed a higher cognitive performance than that of the noncarriers boys. The genetic polymorphism of NLGN4 also had a significant effect on the boys' cognitive ability and other intelligence factors. Future research will involve determining the relationship between NLGN4 and personal cognitive ability.

Positive association of neuroligin-4 gene with nonspecific mental retardation in the Qinba Mountains Region of China.

OBJECTIVE: Neuroligin-4 is essential for proper brain function. Some studies indicate a close relationship between neuroligin-4 and several human psychiatric conditions. METHODS: The case-control method was used to study the association between nonspecific mental retardation (NSMR) and genetic variants of neuroligin-4 gene (NLGN4). Five single nucleotide polymorphisms (SNPs: rs5916271, rs7049300, rs6638575, rs3810686, and rs1882260) were genotyped by PCR-RFLP/SSCP method in the NLGN4. RESULTS: Individual SNP analysis shows significant differences at SNPs rs3810686 and rs1882260 for allele frequency when NSMR cases and controls [odds ratio (OR)=1.589, 95% confidence interval (CI)=1.035-2.438, chi2=4.53, df=1, P=0.033; OR=2.050, 95% CI=1.211-3.470, chi2=7.38, df=1, P=0.007, respectively] were compared. Further haplotype analysis indicates that there are two haplotype sets, rs3810686-rs1882260 and rs6638575-rs3810686-rs1882260, which show statistical differences between NSMR cases and controls (chi2=6.79, df=2, global P=0.034; chi2=9.29, df=2, global P=0.0096, respectively). CONCLUSION: The results suggest a positive association between the genetic variants of the NLGN4 and NSMR in the Chinese children from Qinba Mountains Region.