Adsorption of Oligo-DNA on Magnesium Aluminum-Layered Double-Hydroxide Nanoparticle Surfaces: Mechanistic Implication in Gene Delivery.

Magnesium aluminum-layered double-hydroxide nanoparticles (LDH NPs) are promising drug-delivery vehicles for gene therapy, particularly for siRNA interference; however, the interactions between oligo-DNA and LDH surfaces have not been adequately elucidated. Through a mechanistic study, oligo-DNA initially appears to rapidly bind strongly to the LDH outer surfaces through interactions with their phosphate backbones via ligand exchange with OH- on Mg2+ centers and electrostatic forces with Al3+. These initial interactions might precede diffusion into interlayer spaces, and this knowledge can be used to design better gene therapy delivery systems.

Chemisorption Mechanism of DNA on Mg/Fe Layered Double Hydroxide Nanoparticles: Insights into Engineering Effective SiRNA Delivery Systems.

Layered double hydroxide nanoparticles (LDH NPs) have attracted interest as an effective gene delivery vehicle in biomedicine. Recent advances in clinic trials have demonstrated the efficacy of Mg/Fe LDHs for hyperphosphatemia treatment, but their feasibility for gene delivery has not been systematically evaluated. As a starting point, we aimed to study the interaction between oligo-DNA and Mg/Fe LDH NPs. Our investigation revealed the chemisorption mechanism of DNA on Mg/Fe LDH surfaces, wherein the phosphate backbone of the DNA polymer coordinates with the metal cations of the LDH lattice via the ligand-exchange process. This mechanistic insight may facilitate future gene delivery applications using Mg/Fe LDH NPs.

Dextranase from Arthrobacter oxydans KQ11-1 inhibits biofilm formation by polysaccharide hydrolysis.

Dental plaque is a biofilm of water-soluble and water-insoluble polysaccharides, produced primarily by Streptococcus mutans. Dextranase can inhibit biofilm formation. Here, a dextranase gene from the marine microorganism Arthrobacter oxydans KQ11-1 is described, and cloned and expressed using E. coli DH5α competent cells. The recombinant enzyme was then purified and its properties were characterized. The optimal temperature and pH were determined to be 60°C and 6.5, respectively. High-performance liquid chromatography data show that the final hydrolysis products were glucose, maltose, maltotriose, and maltotetraose. Thus, dextranase can inhibit the adhesive ability of S. mutans. The minimum biofilm inhibition and reduction concentrations (MBIC50 and MBRC50) of dextranase were 2 U ml-1 and 5 U ml-1, respectively. Scanning electron microscopy and confocal laser scanning microscope (CLSM) observations confirmed that dextranase inhibited biofilm formation and removed previously formed biofilms.

Transcriptome and Metabolome Analyses Reveal the Involvement of Multiple Pathways in Flowering Intensity in Mango.

Mango (Mangifera indica L.) is famous for its sweet flavor and aroma. China is one of the major mango-producing countries. Mango is known for variations in flowering intensity that impacts fruit yield and farmers' profitability. In the present study, transcriptome and metabolome analyses of three cultivars with different flowering intensities were performed to preliminarily elucidate their regulatory mechanisms. The transcriptome profiling identified 36,242 genes. The major observation was the differential expression patterns of 334 flowering-related genes among the three mango varieties. The metabolome profiling detected 1,023 metabolites that were grouped into 11 compound classes. Our results show that the interplay of the FLOWERING LOCUS T and CONSTANS together with their upstream/downstream regulators/repressors modulate flowering robustness. We found that both gibberellins and auxins are associated with the flowering intensities of studied mango varieties. Finally, we discuss the roles of sugar biosynthesis and ambient temperature pathways in mango flowering. Overall, this study presents multiple pathways that can be manipulated in mango trees regarding flowering robustness.

Neuromorphic computing chip with spatiotemporal elasticity for multi-intelligent-tasking robots.

Recent advances in artificial intelligence have enhanced the abilities of mobile robots in dealing with complex and dynamic scenarios. However, to enable computationally intensive algorithms to be executed locally in multitask robots with low latency and high efficiency, innovations in computing hardware are required. Here, we report TianjicX, a neuromorphic computing hardware that can support true concurrent execution of multiple cross-computing-paradigm neural network (NN) models with various coordination manners for robotics. With spatiotemporal elasticity, TianjicX can support adaptive allocation of computing resources and scheduling of execution time for each task. Key to this approach is a high-level model, "Rivulet," which bridges the gap between robotic-level requirements and hardware implementations. It abstracts the execution of NN tasks through distribution of static data and streaming of dynamic data to form the basic activity context, adopts time and space slices to achieve elastic resource allocation for each activity, and performs configurable hybrid synchronous-asynchronous grouping. Thereby, Rivulet is capable of supporting independent and interactive execution. Building on Rivulet with hardware design for realizing spatiotemporal elasticity, a 28-nanometer TianjicX neuromorphic chip with event-driven, high parallelism, low latency, and low power was developed. Using a single TianjicX chip and a specially developed compiler stack, we built a multi-intelligent-tasking mobile robot, Tianjicat, to perform a cat-and-mouse game. Multiple tasks, including sound recognition and tracking, object recognition, obstacle avoidance, and decision-making, can be concurrently executed. Compared with NVIDIA Jetson TX2, latency is substantially reduced by 79.09 times, and dynamic power is reduced by 50.66%.

Identification of archaeon-producing hyperthermophilic alpha-amylase and characterization of the alpha-amylase.

The extremely thermophilic anaerobic archaeon strain, HJ21, was isolated from a deep-sea hydrothermal vent, could produce hyperthermophilic alpha-amylase, and later was identified as Thermococcus from morphological, biochemical, and physiological characteristics and the 16S ribosomal RNA gene sequence. The extracellular thermostable alpha-amylase produced by strain HJ21 exhibited maximal activity at pH 5.0. The enzyme was stable in a broad pH range from pH 5.0 to 9.0. The optimal temperature of alpha-amylase was observed at 95 degrees C. The half-life of the enzyme was 5 h at 90 degrees C. Over 40% and 30% of the enzyme activity remained after incubation at 100 degrees C for 2 and 3 h, respectively. The enzyme did not require Ca(2+) for thermostability. This alpha-amylase gene was cloned, and its nucleotide sequence displayed an open reading frame of 1,374 bp, which encodes a protein of 457 amino acids. Analysis of the deduced amino acid sequence revealed that four homologous regions common in amylases were conserved in the HJ21 alpha-amylase. The molecular weight of the mature enzyme was calculated to be 51.4 kDa, which correlated well with the size of the purified enzyme as shown by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Amylase-assisted extraction and antioxidant activity of polysaccharides from Gracilaria lemaneiformis.

Gracilaria lemaneiformis (G. lemaneiformis) is rich in polysaccharides that have many functional activities. In this study, the thermostable α-amylase-assisted extraction and antioxidant activity of water-soluble polysaccharides from G. lemaneiformis were investigated. The extraction conditions were optimized as follows: time, 40 min; temperature, 95 °C; pH, 5 and enzyme amount, 6000 U/g, under which the yield of G. lemaneiformis polysaccharides (GPs) reached 49.15% dry base. The GPs were characterized by monosaccharide composition, FTIR spectrum, UV spectrum and antioxidant activities. The results indicate that the GPs had strong hydroxyl radical activity at a concentration of 100 µg/mL and could be used as potential antioxidants.

Hypoglycaemic and hypolipidaemic properties of peach gum polysaccharides.

Hyperglycaemia and hyperlipidaemia are major risk factors for coronary artery diseases and atherosclerosis. Peach gum polysaccharides (PGPs) possess various bioactivities. In this study, PGPs were extracted with thermostable α-amylase and investigated in terms of hypolipidaemic and hypoglycaemic activities. KKAy mice were gavaged once daily with either PGPs or distilled water (control group) for 3 weeks. Oral administration of PGPs decreased the levels of serum triglyceride, cholesterol low-density lipoprotein cholesterol, fasting blood glucose, plasma insulin, C-peptide, and HbAlc in mice. Moreover, treatment with PGPs increased the insulin sensitivity index in KKAy mice. Results indicated that PGPs possess significant hypoglycaemic effects and could be developed as a drug for preventing hyperlipidaemia and hyperglycaemia.

Antiageing activities of water-soluble chitosan from Clanis bilineata larvae.

Water extracts were prepared using water-soluble chitosan from Clanis bilineata larva skin. This study also investigated antioxidant and antiageing activities of water-soluble chitosan. Water-soluble chitosan of Clanis bilineata larva skin (CBLSWSC) showed considerable scavenging activities on superoxide anion and hydroxyl radicals and reducing capacity. Different doses of CBLSWSC were intragastrically administered on a d-galactose-induced-aged mouse model over a period of six weeks. Intragastric CBLSWSC administration significantly increased activities of superoxide dismutase and glutathione peroxidase and inhibited formation of malondialdehyde in brains and sera of mice in a dose-dependent manner. Results suggest that CBLSWSC exhibits high antioxidant activity and can be developed as potential dietary supplement to retard ageing in humans.

The hyperthermophilic α-amylase from Thermococcus sp. HJ21 does not require exogenous calcium for thermostability because of high-binding affinity to calcium.

The hyperthermophilic α-amylase from Thermococcus sp. HJ21 does not require exogenous calcium ions for thermostability, and is a promising alternative to commercially available α-amylases to increase the efficiency of industrial processes like the liquefaction of starch. We analyzed the amino acid sequence of this α-amylase by sequence alignments and structural modeling, and found that this α-amylase closely resembles the α-amylase from Pyrococcus woesei. The gene of this α-amylase was cloned in Escherichia coli and the recombinant α-amylase was overexpressed and purified with a combined renaturation-purification procedure. We confirmed thermostability and exogenous calcium ion independency of the recombinant α-amylase and further investigated the mechanism of the independency using biochemical approaches. The results suggested that the α-amylase has a high calcium ion binding affinity that traps a calcium ion that would not dissociate at high temperatures, providing a direct explanation as to why the addition of calcium ions is not required for thermostability. Understanding of the mechanism offers a strong base on which to further engineer properties of this α-amylase for better potential applications in industrial processes.

Regenerative nanobots based on magnetic layered double hydroxide for azo dye removal and degradation.

A highly regenerative multifunctional nanobot system, using Fe3O4@SiO2@MgFe-LDH nanoparticles, is developed for efficient removal of waterborne azo dyes and pharmaceuticals. Efficient capture of pollutants, powerful Fenton degradation, and superior materials regeneration lead to a simple and cost-effective wastewater remediation solution.

Chloride-accelerated Cu-Fenton chemistry for biofilm removal.

Biofilms present challenges to numerous industries. Herein, a simple approach was developed based on chloride-accelerated Fenton chemistry, where copper oxide nanoparticles facilitate efficient generation of reactive chlorine species for biofilm removal.

Effect of oligosaccharides derived from Laminaria japonica-incorporated pullulan coatings on preservation of cherry tomatoes.

Laminaria japonica-derived oligosaccharides (LJOs) exhibit antibacterial and antioxidant activities, and pullulan is a food thickener that can form impermeable films. The ability of pullulan coatings with various LJO concentrations (1% pullulan+0.1%, 0.2% or 0.3% LJOs) to preserve cherry tomatoes during storage at room temperature was investigated. The LJO-incorporated pullulan coatings were found to effectively reduce respiratory intensity, vitamin C loss, weight loss and softening, as well as to increase the amount of titratable acid and the overall likeness of fruit compared with the control. These effects were observed to be dose-dependent. Therefore, using LJO-incorporated pullulan coatings can extend the shelf life of cherry tomatoes.

Production of pullulan from raw potato starch hydrolysates by a new strain of Auerobasidium pullulans.

In the present study, hydrolysis of potato starch with marine cold-adapted α-amylase and pullulan production from the hydrolysates by a new strain of Auerobasidium pullulans isolated from sea mud were conducted. The hydrolysis conditions were optimized as follows: reaction time 2h, pH 6.5, temperature 20°C, and α-amylase amount 12 U/g. Under these optimum hydrolysis conditions, the DE value of the potato starch hydrolysates reached to 49.56. The potato starch hydrolysates consist of glucose, maltose, isomaltose, maltotriose, and trace of other maltooligosaccharides with degree of polymerization ranged 4-7. The maximum production of pullulan at 96 h from the hydrolysate of potato starch was 36.17 g/L, which was higher than those obtained from glucose (22.07 g/L, p<0.05) and sucrose (31.42 g/L, p<0.05). Analysis of the high performance liquid chromatography of the hydrolysates of the pullulan product with pullulanase indicated that the main composition is maltotriose, thus confirming the pullulan structure of this pullulan product.

Chloride accelerated Fenton chemistry for the ultrasensitive and selective colorimetric detection of copper.

A highly selective, ultrasensitive (visual and instrumental detection limits of 40 nM and 0.1 nM, respectively), environmentally-friendly, simple and rapid colorimetric sensor was developed for the detection of copper(II) in water. This sensor is based on a novel signal-amplification mechanism involving reactive halide species (RHSs) including chlorides or bromides, which accelerate copper Fenton reactions oxidizing the chromogenic substrate to develop colour. The results of this study expand our understanding of copper-based Fenton chemistry.

Adsorption of doxorubicin on citrate-capped gold nanoparticles: insights into engineering potent chemotherapeutic delivery systems.

Gold nanomaterials have received great interest for their use in cancer theranostic applications over the past two decades. Many gold nanoparticle-based drug delivery system designs rely on adsorbed ligands such as DNA or cleavable linkers to load therapeutic cargo. The heightened research interest was recently demonstrated in the simple design of nanoparticle-drug conjugates wherein drug molecules are directly adsorbed onto the as-synthesized nanoparticle surface. The potent chemotherapeutic, doxorubicin often serves as a model drug for gold nanoparticle-based delivery platforms; however, the specific interaction facilitating adsorption in this system remains understudied. Here, for the first time, we propose empirical and theoretical evidence suggestive of the main adsorption process where (1) hydrophobic forces drive doxorubicin towards the gold nanoparticle surface before (2) cation-π interactions and gold-carbonyl coordination between the drug molecule and the cations on AuNP surface facilitate DOX adsorption. In addition, biologically relevant compounds, such as serum albumin and glutathione, were shown to enhance desorption of loaded drug molecules from AuNP at physiologically relevant concentrations, providing insight into the drug release and in vivo stability of such drug conjugates.

Improving stability of a novel dextran-degrading enzyme from marine Arthrobacter oxydans KQ11.

Dextranases can hydrolyze dextran, so they are used in the sugar industry to mitigate the milling problems associated with dextran contamination. Few studies have been carried out on the storage stability of dextranase, let alone the dextranase of Arthrobacter oxydans KQ11 isolated from sea mud samples. This study improved the storage stability of dextranase from marine A. oxydans KQ11 by adding enzymatic protective reagents (stabilizer and antiseptic). Initially, the conditions (55 °C and 30 min) for maintaining 50% dextranase activity were obtained. Then, the best stabilizers of dextranase were obtained, namely, glycerol (16%), sodium acetate (18%) and sodium citrate (20%). Results showed that p-hydroxybenzoic acid compound sodium acetate (0.05%), D-sodium isoascorbiate (0.03%), and potassium sorbate (0.05%) were the best antiseptics. Subsequent validation experiment showed that dextranase with enzymatic protective reagents maintained 70.8% and 28.96% activities at the 13th week at 25 and 37 °C, respectively.

The atmospheric and room-temperature plasma (ARTP) method on the dextranase activity and structure.

A novel atmospheric and room-temperature plasma (ARTP) method was used to breed high-yielding mutations of Arthrobacter KQ11. Mutagenesis produced two mutations, 4-1 and 4-13, which increased enzyme activity by 19 and 30%, respectively. Dents on the cell envelope were observed under scanning electron microscopy (SEM). The optimal temperature and pH of the wild strain were 45°C and 5.5 and those of the mutant strains were 45°C, pH 6.0 (4-1) and 50°C, pH 6.0 (4-13). Under optimal enzyme production conditions of the wild and mutant strains, the dextranase activity of 4-13 was 50% higher than that of the wild strain. Through amino acid alignment, several nucleotides of the mutant strains were found to have changed. Experiments performed in vitro suggested that this endo-dextranase may inhibit biofilm formation by Streptococcus mutans.

Purification and characterization of a novel marine Arthrobacter oxydans KQ11 dextranase.

Dextranases can hydrolyze dextran deposits and have been used in the sugar industry. Microbial strains which produce dextranases for industrial use are chiefly molds, which present safety issues, and dextranase production from them is impractically long. Thus, marine bacteria to produce dextranases may overcome these problems. Crude dextranase was purified by a combination of ammonium sulfate fractionation and ion-exchange chromatography, and then the enzyme was characterized. The enzyme was 66.2 kDa with an optimal temperature of 50°C and a pH of 7. The enzyme had greater than 60% activity at 60°C for 1h. Moreover, 10mM Co(2+) enhanced dextranase activity (196%), whereas Ni(2+) and Fe(3+) negatively affected activity. 0.02% xylitol and 1% alcohol enhanced activity (132.25% and 110.37%, respectively) whereas 0.05% SDS inhibited activity (14.07%). The thickness of S. mutans and mixed-species oral biofilm decreased from 54,340 nm to 36,670 nm and from 64,260 nm to 43,320 nm, respectively.

A novel method for promoting antioxidant exopolysaccharidess production of Bacillus licheniformis.

A novel method was described for improving the production of antioxidant extracellular polysaccharides from Bacillus licheniformis. Firstly, the tolerances of the strains to the organic solvents were investigated. Wild type strain of B. licheniformis OSTK95 and mutant strain UD061 can grow in a liquid medium in the presence of organic solvents with the logP value equal to or higher than 3.5 and 3.1, respectively. Secondly, the effects of different concentrations of n-hexane and xylene treatment on the extracellular polysaccharides excretion of both strains were studied. The maximum yield of the extracellular polysaccharides of B. licheniformis OSTK95 was 68.59 mg L(-1) after treated by 10% n-hexane or 1% xylene for 3h, while the maximum yield of the extracellular polysaccharides of strain UD061 was 185.01 mg L(-1) after treated by 12.5% n-hexane or 5% xylene for 3h. Finally, the continuous passage experiment showed that the strains have high genetic stability.