Pilot composite tubular bioreactor for outdoor photo-fermentation hydrogen production: From batch to continuous operation.

A novel 70 L composite tubular photo-bioreactor was constructed, and its photo-fermentation hydrogen production characteristics of batch and continuous modes were investigated with glucose as the substrate in an outdoor environment. In the batch fermentation stage, the hydrogen production rate peaked at 37.6 mL H2/(L·h) accompanied by a high hydrogen yield of 7 mol H2/mol glucose. The daytime light conversion efficiency is 4 %, with 37 % of light energy from the sun. An optimal hydraulic retention time of 5 d was identified during continuous photo-fermentation. Under this condition, the stability of the cell concentration is maintained and more electrons can be driven to the hydrogen generation pathway while attaining a hydrogen production rate of 20.7 ± 0.9 mL H2/(L·h). The changes of biomass, volatile fatty acids concentration and ion concentration during fermentation were analyzed. Continuous hydrogen production by composite tubular photo-bioreactor offers new ideas for the large-scale deployment of photobiological hydrogen production.

Double-Pulse Ultrasonic Welding of Carbon-Fiber-Reinforced Polyamide 66 Composite.

Ultrasonic welding of thermoplastics is widely applied in automobile and aerospace industries. Increasing the weld area and avoiding thermal decomposition are contradictory factors in improving strength of ultrasonically welded polymers. In this study, relations among the loss modulus of carbon-fiber-reinforced polyamide 66 composite (CF/PA 66), time for obtaining stable weld area, and time for CF/PA 66 decomposition are investigated systematically. Then, a double-pulse ultrasonic welding process (DPUW) is proposed, and the temperature evolutions, morphologies and structures of fractured surfaces, and tensile and fatigue properties of the DPUWed joints are measured and assessed. Experimental results show the optimal welding parameters for DPUW include a weld time of 2.1 s for the first pulse, a cooling time of 12 s, and a weld time of 1.5 s for the second pulse. The DPUW process enlarged the weld area while avoided decomposition of CF/PA 66 under appropriate welding parameters. Compared to the single-pulse welded joint, the peak load, weld area, and endurance limit of the DPUWed joint increased by about 15%, 23% and 59%, respectively. DPUW also decreases the variance in strengths of the joints.

High-Performance HVOF-Sprayed Fe-Based Amorphous Coating on LA141 Magnesium Alloy via Optimizing Oxygen Flow and Kerosene Flow.

To solve the problem of poor corrosion and wear resistance of Mg-Li alloys, Fe-based amorphous coatings were prepared by high velocity oxygen-fuel spraying technology (HVOF) on the LA141 magnesium alloy substrate with a Ni60 intermediate layer. The microstructure and performance of Fe-based amorphous coatings with different oxygen flow and kerosene flow were characterized and analyzed. The results demonstrate that there is an optimal oxygen/kerosene ratio where the porosity of Fe-based amorphous coating is the lowest. Moreover, the amorphous content increases with the decrease in the oxygen/kerosene ratio. In particular, when the oxygen flow is 53.8 m3/h and the kerosene flow is 26.5 L/h, the Fe-based amorphous coating possesses the lowest porosity (0.87%), the highest hardness (801 HV0.1), the highest bonding strength (56.9 MPa), and an excellent corrosion and wear resistance. Additionally, it can be seen that the Fe-based amorphous coating is composed of amorphous splats and amorphous oxides, but the Ni60 intermediate layer exhibits an amorphous and crystalline multi-phase structure. The high bonding strength of the coating is attributed to the low porosity of Fe-based amorphous coating and the localized metallurgical bonding between different layers. Finally, the mechanisms on corrosion and wear of Fe-based amorphous coatings are also discussed.

Identification of Genes Involved in Fe-S Cluster Biosynthesis of Nitrogenase in Paenibacillus polymyxa WLY78.

NifS and NifU (encoded by nifS and nifU) are generally dedicated to biogenesis of the nitrogenase Fe-S cluster in diazotrophs. However, nifS and nifU are not found in N2-fixing Paenibacillus strains, and the mechanisms involved in Fe-S cluster biosynthesis of nitrogenase is not clear. Here, we found that the genome of Paenibacillus polymyxa WLY78 contains the complete sufCDSUB operon, a partial sufC2D2B2 operon, a nifS-like gene, two nifU-like genes (nfuA-like and yutI), and two iscS genes. Deletion and complementation studies showed that the sufC, sufD, and sufB genes of the sufCDSUB operon, and nifS-like and yutI genes were involved in the Fe-S cluster biosynthesis of nitrogenase. Heterologous complementation studies demonstrated that the nifS-like gene of P. polymyxa WLY78 is interchangeable with Klebsiella oxytoca nifS, but P. polymyxa WLY78 SufCDB cannot be functionally replaced by K. oxytoca NifU. In addition, K. oxytoca nifU and Escherichia coli nfuA are able to complement the P. polymyxa WLY78 yutI mutant. Our findings thus indicate that the NifS-like and SufCDB proteins are the specific sulfur donor and the molecular scaffold, respectively, for the Fe-S cluster formation of nitrogenase in P. polymyxa WLY78. YutI can be an Fe-S cluster carrier involved in nitrogenase maturation in P. polymyxa WLY78.

cpubi4 Is Essential for Development and Virulence in Chestnut Blight Fungus.

Ubiquitination plays key roles in eukaryotic growth, stress adaptation, and metabolic regulation. In our previous work, ubiquitin was found to be secreted in the hypovirus-infected strain of Cryphonectria parasitica, a phytopathogenic filamentous fungus responsible for the chestnut blight. Here we report the functional and molecular characterization of a polyubiquitin gene, cpubi4, in C. parasitica. The expression of cpubi4 was upregulated by the infection of a hypovirus. Deletion of cpubi4 resulted in abnormal morphology, reduced sporulation, attenuation of virulence, and significant reduction in ubiquitination. A total of 378 sites in 236 proteins were identified to be significantly decreased in ubiquitination in the absence of cpubi4. Quantitative proteome analysis revealed that 285 in 4,776 identified proteins changed in abundance (1.5-fold, P < 0.05) in the cpubi4 null mutant, as compared with the wild-type strain.

Comparative Methylome Analysis Reveals Perturbation of Host Epigenome in Chestnut Blight Fungus by a Hypovirus.

In eukaryotic genomes, DNA methylation is an important type of epigenetic modification that plays crucial roles in many biological processes. To investigate the impact of a hypovirus infection on the methylome of Cryphonectria parasitica, the chestnut blight fungus, whole-genome bisulfite sequencing (WGBS) was employed to generate single-base resolution methylomes of the fungus with/without hypovirus infection. The results showed that hypovirus infection alters methylation in all three contexts (CG, CHG, and CHH), especially in gene promoters. A total of 600 differentially methylated regions (DMRs) were identified, of which 144 could be annotated to functional genes. RNA-seq analysis revealed that DNA methylation in promoter is negatively correlated with gene expression. Among DMRs, four genes were shown to be involved in conidiation, orange pigment production, and virulence. Taken together, our DNA methylomes analysis provide valuable insights into the understanding of the relationship between DNA methylation and hypovirus infection, as well as phenotypic traits in C. parasitica.

Biodegradable Transparent Substrate Based on Edible Starch-Chitosan Embedded with Nature-Inspired Three-Dimensionally Interconnected Conductive Nanocomposites for Wearable Green Electronics.

Electronic waste (E-waste) contain large environmental contaminants such as toxic heavy metals and hazardous chemicals. These contaminants would migrate into drinking water or food chains and pose a serious threat to environment and human health. Biodegradable green electronics has great potential to address the issue of E-waste. Here, we report on a novel biodegradable and flexible transparent electrode, integrating three-dimensionally (3D) interconnected conductive nanocomposites into edible starch-chitosan-based substrates. Starch and chitosan are extracted from abundant and inexpensive potato and crab shells, respectively. Nacre-inspired interface designs are introduced to construct a 3D interconnected single wall carbon nanotube (SCNT)-pristine graphene (PG)-conductive polymer network architecture. The inorganic one-dimensional SCNT and two-dimensional PG sheets are tightly cross-linked together at the junction interface by long organic conductive poly(3,4-ethylenedioxythiophene) (PEDOT) chains. The formation of a 3D continuous SCNT-PG-PEDOT conductive network leads to not only a low sheet resistance but also a superior flexibility. The flexible transparent electrode possesses an excellent optoelectronic performance: typically, a sheet resistance of 46 Ω/sq with a transmittance of 83.5% at a typical wavelength of 550 nm. The sheet resistance of the electrode slightly increased less than 3% even after hundreds of bending cycles. The lightweight flexible and biocompatible transparent electrode could conform to skin topography or any other arbitrary surface naturally. The edible starch-chitosan substrate-based transparent electrodes could be biodegraded in lysozyme solution rapidly at room temperature without producing any toxic residues. SCNT-PG-PEDOT can be recycled via a membrane process for further fabrication of conductive and reinforcement composites. This high-performance biodegradable transparent electrode is a promising material for next-generation wearable green optoelectronics, transient electronics, and edible electronics.

Targeting ETS1 with RNAi-based supramolecular nanoassemblies for multidrug-resistant breast cancer therapy.

Overexpression of erythroblastosis virus E26 oncogene homolog 1 (ETS1) gene is correlated with both tumor progression and poor response to chemotherapy in cancer treatment, and the exploitation of RNA interference (RNAi) technology to downregulate ETS1 seems to be a promising approach to reverse multidrug-resistant cancer cells to chemotherapy. Hence, the RNAi-based nanomedicine which is able to simultaneously downregulate ETS1 expression and to deliver chemotherapeutic agents may improve multidrug-resistant cancer therapy synergistically. In this study, we developed a supramolecular nanoassembly that could deliver siRNA targeting ETS1 (siETS1) and doxorubicin (DOX) as an effective nanomedicine to achieve successful chemotherapy towards multidrug-resistant breast cancer. The nanotherapeutic system was prepared by loading adamantane-conjugated doxorubicin (AD) into polyethyleneimine-modified (2-hydroxypropyl)-γ-cyclodextrin (HP) through the supramolecular assembly to form AD-loaded HP (HPAD), followed by electrostatically-driven self-assembly between siETS1 and HPAD. When the HPAD/siETS1 nanoassemblies were delivered into drug-resistant MCF-7/ADR cells, the drug efflux was significantly reduced as a result of simultaneous silencing of ETS1 and MDR1 genes. Importantly, the HPAD/siETS1 nanoassembly could enhance drug residence time at tumor site, and effectively inhibit drug-resistant tumor growth due to the inhibition of angiogenesis and necrosis in tumor tissues. Western blot analysis indicated that the gene expression of both ETS1 and MDR1 in vivo was considerably downregulated after the drug-resistant tumor-bearing mouse was treated with HPAD/siETS1 nanoassemblies. This study offers a new therapeutic delivery strategy targeting ETS1 for the effective multidrug-resistant chemotherapy.

Restoration of chemosensitivity by multifunctional micelles mediated by P-gp siRNA to reverse MDR.

One of the main obstacles in tumor therapy is multiple drug resistance (MDR) and an underlying mechanism of MDR is up-regulation of the transmembrane ATP-binding cassette (ABC) transporter proteins, especially P-glycoprotein (P-gp). In the synergistic treatment of siRNA and anti-cancer drug doxorubicin, it is crucial that both the siRNA and doxorubicin are simultaneously delivered to the tumor cells and the siRNA can fleetly down-regulate P-g before doxorubicin inactivates the P-gp and is pumped out. Herein, a type of micelles comprising a polycationic PEI-CyD shell to condense the siRNA and hydrophobic core to package doxorubicin is reported. The structure of the polymer is determined by (1)H NMR, FT-IR, DSC, and XRD and the micelles are characterized by DLS, 2D-NOESY NMR, and TEM to study the self-assembly of the micelles with siRNA and drugs. In vitro studies demonstrate controlled release and temporal enhancement of the therapeutic efficacy of P-gp siRNA and doxorubicin. Release of siRNA down-regulates the mRNA and protein levels of P-gp in the MCF-7/ADR cell lines effectively and the accumulated doxorubicin facilitates apoptosis of the cells to reverse MDR. Moreover, in vivo research reveals that the siRNA and doxorubicin loaded micelles induce tumor cell apoptosis and inhibit the growth of MDR tumor. The western blotting and RT-PCR results illustrate that the synergistic treatment of siRNA and doxorubicin leads to efficient reduction of the P-gp expression as well as cell apoptotic induction in MDR tumors at a small dosage of 0.5 mg/kg. The micelles have large clinical potential in drug/RNAi synergistic treatment via restoration of the chemosensitivity in MDR cancer therapy.

Quantitative analysis and pharmacokinetics study of integrin antagonist AP25 in rat plasma.

BALB/c mice were immunized by highly immunogenic recombinant proteins containing amino acid sequence of integrin antagonist AP25. Antibody against AP25 was prepared and purified by affinity chromatography. An indirect enzyme-linked immunosorbent assay for qualitative analysis of AP25 in rat plasma samples was successfully established. The assay was successfully applied to determine the pharmacokinetic parameters of AP25 in SD rats by intravenous administration of AP25 and then the rat plasma protein binding of AP25 were determined in vitro. In order to investigate the specificity of ELISA for detection of prototype AP25 in plasma, the proportion of AP25 prototype drug in the ELISA signal value was validated by HPLC. These results can serve as valuable future clinical trials.

RGD-modified endostatin fragments showed an antitumor effect through antiangiogenesis.

EDSM, an endostatin-derived synthetic polypeptide, contains the amino acids 6-48 of endostatin from its N-terminus, which could inhibit human umbilical vein endothelial cell (HUVEC) migration and tumor growth. To increase the targeted delivery of EDSM to tumors and further enhance its antiangiogenic activity, the RGD sequence (Arg-Gly-Asp) was introduced into EDSM and two peptides were obtained: EDSM-X with RGD on the N-terminus of EDSM and EDSM-Y with RGD on the C-terminus. Both modified peptides showed a significant antiangiogenic activity in the HUVEC migration assay, the HUVEC tube formation assay, and the murine aortic ring formation assay in vitro. In agreement with the in-vitro data, EDSM-X and EDSM-Y also showed a significant antitumor activity in vivo. From the cell adhesion assay, it was confirmed that the molecular target of the modified peptides on HUVECs was integrin αvß3, rather than integrin α5ß1. Furthermore, EDSM-Y exhibited more potent antiangiogenic activity and antitumor activity than EDSM-X in vitro and in vivo, and this phenomenon was attributed to the difference in the two modified peptides in their three-dimensional structure modeling and their molecular dockings with integrin αvß3.

In vivo pharmacokinetics, immunogenicity and mechanism of PEGylated antitumor polypeptide.

HM-3, An RGD (Arg-Gly-Asp)-modified polypeptide derived from endostatin, is a potent angiogenesis inhibitor. Its robust inhibitory effects on endothelial cell migration and tumor growth have been demonstrated by activity assay both in vitro and in vivo. However, HM-3 has relatively short half-life in vivo. In order to prolong its half-life and retain its safety and efficacy, previous studies modified HM-3 with four types of PEG (site-specific N-terminal modification), and the results showed that mPEG-SC20k-HM-3 was the most ideal modification product via activity evaluation in vivo. In the present study, we determined the pharmacokinetic properties, immunogenicity and binding targets of mPEG-SC20k-HM-3. The results showed that mPEG-SC20k-HM-3 had good linear pharmacokinetic properties in SD rats. The half-life of mPEG-SC20k-HM-3 was 43.76-fold longer than that of unmodified HM-3 after intravenous injection in SD rats. The administration frequency of the modified product (mPEG-SC20k-HM-3) was reduced from twice a day to once every 2 days, while the safety and efficacy were retained. The immunogenicity of mPEG-SC20k-HM-3 was significantly lower than that of HM-3 in BALB/c mice. Histochemical and immunohistochemical results showed that mPEG-SC20k-HM-3 could significantly inhibit angiogenesis and tumor growth, induce continuous necrosis, and reduce vessel density within tumor tissues. Furthermore, mPEG-SC20k-HM-3 could bind multi-target αvß3 and α5ß1 of integrin, and the major binding target was integrin αvß3. All of these results indicated that PEGylated HM-3 had a good application prospect.

[Chemical components and their allelopathic effects of the volatiles from Larix principis-rupprechtii leaves and branches].

The chemical components of the volatiles from the leaves and branches of Larix principis-rupprechtii were analyzed by GC-MS, with their allelopathic effects on the seed germination and growth of L. principis-rupprechtii itself investigated. The results indicated that the volatiles contained 20 major components, and the main constituents were terpenes and their oxidized derivatives, in which, there were 5 monoterpenes and 12 sesquiterpenes. The volatiles from the leaves and branches of L. principis-rupprechtii had significant inhibitory effects on the germination rate, germination speed index, radical length, hypocotyl length, and fresh mass of L. principis-rupprechtii itself, indicating that these volatiles contained potential allelochemicals, which might be related to the higher contents of terpenes and their oxidized derivatives. The allelopathic effects of the volatiles affected the regeneration of L. principis-rupprechtii, which should be a reason for the degeneration of artificial pure L. principis-rupprechti forest.