A Coenzyme-Free Biocatalyst for the Value-Added Utilization of Lignin-Derived Aromatics.

Value-added utilization of lignin waste streams is vital to fully sustainable and economically viable biorefineries. However, deriving substantial value from its main constituents is seriously hindered by the constant requirement for expensive coenzymes. Herein, we devised a coenzyme-free biocatalyst that could transform lignin-derived aromatics into various attractive pharmaceutical and polymer building blocks. At the center of our strategy is the integrated use of new mining phenolic acid decarboxylase and aromatic dioxygenase with extremely high catalytic efficiency, which realizes the value-added utilization of lignin in a coenzyme-independent manner. Notably, a new temperature/pH-directed strategy was proposed to eliminate the highly redundant activities of endogenous alcohol dehydrogenases. The major components of lignin were simultaneously converted to vanillin and 4-vinylphenol. Since the versatile biocatalyst could efficiently convert many other renewable lignin-related aromatics to valuable chemicals, this green route paves the way for enhancing the entire efficiency of biorefineries.

Optimization and continuous fabrication of moth-eye nanostructure array on flexible polyethylene terephthalate substrate towards broadband antireflection.

Reflection loss can cause harmful effects on the performance of optoelectronic devices, such as cell phones, notebooks, displays, solar cells, and light-emitting diode (LED) devices. In order to obtain broadband antireflection (AR) properties, many researchers have utilized surface texture techniques to produce AR subwavelength structures on the interfaces. Among the AR subwavelength structures, the moth-eye nanostructure is one of the most promising structures, with the potential for commercialization in the near future. In this research, to obtain broadband AR performance, the optimization of moth-eye nanostructures was first carried out using the finite difference time domain method within the spectral ranges of 400-800 nm, including the optimization of shape, height, pitch, and residual layer thickness. In addition, the continuous production of moth-eye nanostructure array upon a flexible polyethylene terephthalate substrate was demonstrated by using the roll-to-roll ultraviolet nanoimprint lithography (R2R UV-NIL) process and anodic aluminum oxide mold, which provided a solution for the cost-effective fabrication of moth-eye nanostructure array. The AR performance of moth-eye nanostructure array obtained by the R2R UV-NIL process was also investigated experimentally, and good consistence was shown with the simulated results. This research can provide a beneficial direction for the optimization and cost-effective production of the moth-eye nanostructure array.

Ligand-directed stearic acid grafted chitosan micelles to increase therapeutic efficacy in hepatic cancer.

Targeted delivery system would be an interesting platform to enhance the therapeutic effect and to reduce the side effects of anticancer drugs. In this study, we have developed lactobionic acid (LA)-modified chitosan-stearic acid (CS-SA) (CSS-LA) to deliver doxorubicin (DOX) to hepatic cancer cells. The average particle size of CSS-LA/DOX was ∼100 nm with a high entrapment efficiency of >95%. Drug release studies showed that DOX release from pH-sensitive micelles is significantly faster at pH 5.0 than at pH 7.4. The LA conjugated micelles showed enhanced cellular uptake in HepG2 and BEL-7402 liver cancer cells than free drug and unconjugated micelles. Consistently, CSS-LA/DOX showed enhanced cell cytotoxicity in these two cell lines. Annexin-V/FITC and PI based apoptosis assay showed that the number of living cells greatly reduced in this group with marked presence of necrotic and apoptotic cells. LA-conjugated carrier induced typical chromatic condensation of cells; membrane blebbing and apoptotic bodies began to appear. In vivo, CSS-LA/DOX showed an excellent tumor regression profile with no toxic side effects. The active targeting moiety, long circulation profile, and EPR effect contributed to its superior anticancer effect in HepG2 based tumor. Our results showed that polymeric micelles conjugated with LA increased the therapeutic availability of DOX in the liver cancer cell based solid tumor without any toxic side effects. The active targeting ligand conjugated nanoparticulate system could be a promising therapeutic strategy in the treatment of hepatic cancers.

Role of automated functional imaging and myocardial work in assessment of cardiac function in children with obstructive sleep apnea.

BACKGROUND: Early identification of abnormal left ventricular function in children with obstructive sleep apnea (OSA) is difficult using conventional echocardiographic indices and commonly used clinical markers of myocardial damage. We sought to investigate the value of automatic function imaging and myocardial work parameters in predicting early cardiac impairment in children having OSA with preserved left heart function and thereby identifying an optimal index for assessment. PATIENTS AND METHODS: Fifty-two children who presented with symptoms of nocturnal sleep snoring and open-mouth breathing and 34 healthy controls were enrolled in this study. Clinical characteristics and conventional echocardiographic data were collected, and image analysis was performed using two-dimensional speckle-tracking echocardiography to obtain left ventricular global longitudinal strain (GLS), post-systolic index, peak strain dispersion, global work index (GWI), global constructive work (GCW), global wasted work, and global work efficiency. RESULTS: Children with OSA had significantly lower GLS, GWI, and GCW than those without (P < 0.05). Additionally, GWI (ß = -32.87, 95% CI: -53.47 to -12.27), and GCW (ß = -35.09, 95% CI: -55.35 to -14.84) were found to correlate with the disease severity in the multiple linear regression mode, with worsening values observed as the severity of the disease increased. ROC curve analysis revealed that GCW was the best predictor of myocardial dysfunction, with an AUC of 0.809 (P < 0.001), and the best cutoff point for diagnosing myocardial damage in children with OSA was 1965.5 mmHg%, with a sensitivity of 92.5% and a specificity of 58.7%. CONCLUSIONS: GLS, GWI, and GCW were identified as predictors of myocardial dysfunction in children with OSA, with GCW being the best predictor.

Establishment of a salt-induced bioremediation platform from marine Vibrio natriegens.

Industrial wastewater discharge, agricultural production, marine shipping, oil extraction, and other activities have caused serious marine pollution, including microplastics, petroleum and its products, heavy metals, pesticides, and other organics. Efficiency of bioremediation of marine pollutions may be limited by high salt concentrations (>1%, w/v), which can cause an apparent loss of microbial activities. In this study, functional promoters P1, P2-1, and P2-2 censoring salt stress were isolated and identified from a Vibrio natriegens strain Vmax. Three salt-induced degradation models were constructed to degrade polyethylene terephthalate (PET), chlorpyrifos (CP), and hexabromocyclododecanes (HBCDs) using the marine strain Vmax. The engineered strains are efficient for degradation of the corresponding substrates, with the degradation rates at 15 mg/L PET in 8 d, 50 mg/L CP in 24 h, and 1 mg/L HBCDs in 4 h, respectively. In addition, an immobilization strategy for recycling and reusing of engineered strains was realized by expressing the chitin-binding protein GbpA. This study may help answer the usage of rapidly growing marine bacteria such as V. natriegens Vmax to degrade marine pollution efficiently.

[Advances of polymer-monomer production by cyanobacterial cell factory].

Cyanobacteria is one of the promising microbial chassis in synthetic biology, which serves as a typical host for light-driven production. With the gradual depletion of fossil resources and intensification of global warming, the research on cyanobacterial cell factory using CO2 as carbon resource is ushering in a new wave. For a long time, research focus on cyanobacterial cell factory has mainly been the production of energy products, such as liquid fuels and hydrogen. One of the critical bottlenecks occurring in cyanobacterial cell factory is the poor economic performance, which is mainly caused by the inherent inefficiency of cyanobacteria. The problem is particularly prominent for these extremely cost-sensitive energy products. As an indispensable basis for modern industry, polymer monomers belong to the bulk chemicals with high added value. Therefore, increasing attention has been focused on polymer monomers which are superior in overcoming the economic barrier in commercialization of cyanobacterial cell factories. Here, we systematically review the progress on the production of polymer monomers using cyanobacteria, including the strategies for improving production, and the related technologies for the application of this important microbial cell factory. Finally, we summarize several issues in cyanobacterial synthetic biology and proposed future developing trends in this field.

[Measurement and analysis of inclining implant in edentulous maxillary surgery design based on cone-beam CT].

PURPOSE: To study the height of alveolar ridge and the distance between the wall of maxillary sinus on the curve of the dental arch, and to explore the guiding significance for maxillary sinus surgery and edentulous jaw implant surgery. METHODS: Cone-beam CT(CBCT) data of 105 objects were selected to measure the height of alveolar ridge and the maximum bone internal distance between the wall of maxillary sinus 10 mm and 12 mm above the alveolar crest horizontally on the curve of the dental arch. Forty-nine objects with alveolar ridge height on the molar area less than or equal to 10 mm were included for further investigation. The distance between the maxillary sinus wall, and the angle between the maxillary sinus wall on the horizontal plane were measured and calculated. SPSS 22.0 software package was used for statistical analysis of the data. RESULTS: The mean value of the alveolar ridge height in the posterior teeth area was first premolar > second premolars > second molars > first molars. The mean value of the horizontal distance between the wall of sinus 10 mm and 12 mm above the alveolar crest horizontally was (82.40±27.56) mm and (70.54±29.70) mm in 105 objects, while (67.85±28.53) mm and (52.75±24.90) mm in 49 included objects, respectively. The mean value of the angle between the maxillary sinus wall on the horizontal plane was (23.55±39.13)°. There was no significant difference between males and females. However, the horizontal distance had statistically significant differences between different groups and in different horizontal levels of the same group. CONCLUSIONS: CBCT is helpful to establish implant surgical plan, and carry out maxillary sinus lift and edentulous jaw implant surgery.

Steps Toward High-Performance PLA: Economical Production of d-Lactate Enabled by a Newly Isolated Sporolactobacillus terrae Strain.

Optically pure d-lactate production has received much attention for its critical role in high-performance polylactic acid production. However, the current technology can hardly meet the comprehensive demand of industrialization on final titer, productivity, optical purity, and raw material costs. Here, an efficient d-lactate producer strain, Sporolactobacillus terrae (S. terrae) HKM-1, is isolated for d-lactate production. The strain HKM-1 shows extremely high d-lactate fermentative capability by using peanut meal, soybean meal, or corn steep liquor powder as a sole nitrogen source; the final titers (205.7 g L-1 , 218.9 g L-1 , and 193.9 g L-1 , respectively) and productivities (5.56 g L-1 h-1 , 5.34 g L-1 h-1 , and 3.73 g L-1 h-1 , respectively) of d-lactate reached the highest level ever reported. A comparative genomic analysis between S. terrae HKM-1 and previously reported d-lactate high-producing Sporolactobacillus inulinus (S. inulinus) CASD is conducted. The results show that many unrelated genetic features may contribute to the superior performance in d-lactate production of S. terrae HKM-1. This d-lactate producer HKM-1, along with its fermentation process, is promising for sustainable d-lactate production by using agro-industrial wastes.

Identification and characterization of a novel prespheroid 3-dimensional hepatocyte monolayer on galactosylated substratum.

Three-dimensional (3D) hepatocyte spheroids mimicking the structural and functional characteristics of hepatocytes in vivo were self-assembled onto a galactosylated polyethylene terephthalate (PET) substratum, and the dynamic process of spheroid formation was investigated using time-lapse confocal microscopy. Hepatocytes cultured on this galactosylated substratum formed small cell-aggregates within 12 h, which gradually merged into "island-like" clusters at approximately 1 day and spread to form prespheroid monolayer within 2 days; the prespheroid monolayer was stretched to fold into compact and larger 3D spheroids after 3 days. We compared the expressions of F-actin (cytoskeleton), phosphorylated focal adhesion kinase (p-FAK, cell-substratum interactions) and E-cadherin (cell-cell interactions) during the dynamic process of 3D hepatocyte spheroid formation with the dynamic process of 2D hepatocyte monolayer formation on collagen substratum. Hepatocytes in the prespheroid monolayer stage exhibited the strongest cell-substratum interactions of all 4 stages during spheroid formation with cell-cell interactions and F-actin distribution comparable with those of the 3D hepatocyte spheroids. The prespheroid monolayer also exhibited better hepatocyte polarity (multidrug resistance protein 2) and tight junction (zonula occludens-1) formation, more-differentiated hepatocyte functions (albumin production and cytochrome P450 1 A activity), and higher sensitivity to hepatotoxicity than the conventional 2D hepatocyte monolayer. The transient prespheroid 3D monolayer could be stabilized on a hybrid glycine-arginine-glycine-aspartic acid-serine (GRGDS)/galactose-PET substratum for up to 1 week and destabilized to form 3D spheroids in excess soluble GRGDS peptide.

Development and validation of a sensitive UPLC-MS/MS instrumentation and alkaline nitrobenzene oxidation method for the determination of lignin monomers in wheat straw.

A method to determine the lignin monomers (p-hydroxybenzaldehyde, vanillin and syringaldehyde) in plant cell wall of wheat internode was developed and validated using a high-throughput nitrobenzene oxidation step and ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) for quantification. UPLC analyses were carried out using an reversed phase C18 column (ACQUITY UPLC BEH, 1.7µm, 2.1×100mm) and gradient elution with water and acetonitrile. This method was completely validated in terms of analyzing speed, linearity, sensitivity, limits of detection (LODs) and limits of quantification (LOQs).The three lignin monomers were successfully separated within 6min and only 2min were required to regain its equilibrium. The method linearity with regression coefficients values (R2) greater than 0.997. Additionally, LODs ranged from 0.21 to 0.89µgL-1 and LOQs ranged from 0.69 to 2.95µgL-1. The applicability of this analytical approach for determining the three lignin monomers was confirmed by the successful analysis of real samples of wheat stem internodes. The nitrobenzene oxidation method was used for the analysis of lignin monomers. We have optimized the treatment temperature (170°C, 1h) and realized the high-throughput using the microwave digestion instrument. Recovery of this extraction method ranged from 68.4% to 77.7%. The analysis result showed that the guaiacyl unit (G) was the major component of lignin and there was a higher content of the syringyl unit (S) than that of the hydroxybenzyl unit (H).

Manipulation of lignin metabolism by plant densities and its relationship with lodging resistance in wheat.

Increasing plant density is one of the most efficient ways of increasing wheat (Triticum aestivum L.) grain production. However, overly dense plant populations have an increased risk of lodging. We examined lignin deposition during wheat stem development and the regulatory effects of plant density using the wheat cultivars shannong23 and weimai8. Plants were cultivated at densities of 75, 225 and 375 plants per m2 during two growing seasons. Our results showed that decreasing plant density enhanced culm quality, as revealed by increased culm diameter, wall thickness and dry weight per unit length, and improved the structure of sclerenchyma and vascular bundles by increasing lignification. In addition, more lignins were deposited in the secondary cell walls, resulting in strong lodging resistance. The guaiacyl unit was the major component of lignin and there was a higher content of the syringyl unit than that of the hydroxybenzyl unit. Furthermore, we hypothesised that the syringyl unit may correlate with stem stiffness. We describe here, to the best of our knowledge, the systematic study of the mechanism involved in the regulation of stem breaking strength by plant density, particularly the effect of plant density on lignin biosynthesis and its relationship with lodging resistance in wheat.

Simultaneous qualitation and quantitation of natural trans-1,4-polyisoprene from Eucommia ulmoides Oliver by gel permeation chromatography (GPC).

Natural trans-1,4-polyisoprene (TPI) as a functional biomaterial has aroused great interest for rubber industrial product use. Here, we proposed a method that enables simultaneous analysis of the content and molecular-weight distribution (MWD) of natural TPI by gel permeation chromatography (GPC). The natural TPIs were collected from leaves, fruit coatings and bark of Eucommia ulmoides Oliver (E. ulmoides) through toluene extraction followed by ethanol purification. The results of TPI contents from leaves and fruit coatings were shown ca. 3.5% and 13.8%, respectively. Accordingly, limits of detection (LODs) of TPI were 0.58mg/mL from leaves and 0.47mg/mL from fruit coatings. The MWDs of TPI demonstrated a bimodal distribution from leaves, a unimodal distribution from bark, and a unimodal distribution with a tiny peak shoulder from fruit coatings. In real-life E. ulmoides analysis, the results from three independent methods (GPC, gravimetric method, and infrared spectroscopy) were obtained with good consistency.

Design improvement and failure reduction of endodontic files through finite element analysis: application to V-Taper file designs.

INTRODUCTION: Torsional stiffness and bending flexibility are essential characteristics as far as the performance and safety of the endodontic files are concerned. Inadequacy in addressing these requirements in file design leads to increased risk of file failure. The stiffness and flexibility of the endodontic file are greatly dependent on its geometric design. The aim of this study was to evaluate the influence of geometric features on the mechanical performance of endodontic files through numerical simulations. METHODS: Finite element models of V-Taper file were developed, and the mechanical behavior of the file under bending and torsional loads was simulated. The influence of helix angle, taper, and flute length was evaluated through parametric studies. RESULTS: In the helix angle range between 5 and 40 degrees, the bending flexibility and torsional stiffness both improve with increasing helix angle. The torsional stiffness increases with increasing taper or decreasing flute length, accompanied by a decrease in bending flexibility. Changing the flute length alone does not result in a change in the stress profile in the tip section. The elastic limit of V-Taper file tip section was estimated in the form of transverse deflection and angular deformation in bending and in torsion, respectively. CONCLUSIONS: The influence of helix angle, taper, and flute length on the bending flexibility and torsional stiffness of V-Taper files was quantitatively assessed through parametric studies with finite element method. The elastic limit of the V-Taper file tip section was estimated. A design methodology for achieving improved mechanical performances was proposed.