ALBERT

Description: Clostridium acetobutylicum has been considered as an attractive platform host for biorefinery due to its metabolic diversity. Considering its capability to overproduce butanol through butyrate, it was thought that butyric acid can also be efficiently produced by this bacterium through metabolic engineering. The pta-ctfB -deficient C. acetobutylicum CEKW, in which genes encoding phosphotransacetylase and CoA-transferase were knocked out, was assessed for its potential as a butyric acid producer in fermentations with four controlled pH values at 5.0, 5.5, 6.0, and 6.4. Butyric acid could be best produced by fermentation of the CEKW at pH 6.0, resulting in the highest titer of 26.6 g/l, which is 6.4 times higher than that obtained with the wild type. However, due to the remaining solventogenic ability of the CEKW, 3.6 g/l solvents were also produced. Thus, the CEKW was further engineered by knocking out the adhE1 -encoding aldehyde/alcohol dehydrogenase to prevent solvent production. Batch fermentation of the resulting C. acetobutylicum HCEKW at pH 6.0 showed increased butyric acid production to 30.8 g/l with a ratio of butyric-to-acetic acid (BA/AA) of 6.6 g/g and a productivity of 0.72 g/l/h from 86.9 g/l glucose, while negligible solvent (0.8 g/l ethanol only) was produced. The butyric acid titer, BA/AA ratio, and productivity obtained in this study were the highest values reported for C. acetobutylicum , and the BA/AA ratio and productivity were also comparable to those of native butyric acid producer Clostridium tyrobutyricum . These results suggested that the simultaneous deletion of the pta-ctfB-adhE1 in C. acetobutylicum resulted in metabolic switch from biphasic to acidogenic fermentation, which enhanced butyric acid production.

Description: In this paper, we have designed, fabricated and characterized silicon nitride grating couplers with high efficiency at 1490 nm. The devices are fabricated using deep UV photolithography with resolution requirement of ∼500 nm. The grating coupler fabricated yields a peak coupling efficiency of −5.1 dB. The 1-dB bandwidth of the grating coupler is 60 nm.

Description: Disruption of spatiotemporal behavior of intracellular signaling cascades including tumor necrosis factor alpha-related apoptosis-inducing ligand (TRAIL)-mediated signaling in prostate cancer has gained tremendous attention in the past few years. There is an increasing effort in translating the emerging information about TRAIL-mediated signaling obtained through experimental and preclinical data to clinic. Fascinatingly, novel targeting approaches are being developed to enhance the tissue- or subcellular-specific delivery of drugs with considerable focus on prostate cancer. These applications have the potential to revolutionize prostate cancer therapeutic strategies and include the accumulation of drugs in target tissue as well as the selection of internalizing ligands for enhanced receptor-mediated uptake of drugs. In this mini-review, we outline outstanding developments in therapeutic strategies based on the regulation and/or targeting of TRAIL pathway for the treatment of prostate cancer. Moreover, microRNAs (miRNAs), with potential transcriptional and posttranscriptional regulation of gene expression, will be presented for their potential in prostate cancer treatment. Emphasis has been given to the use of delivery approaches, especially based on nanotechnology. Considerably, enhanced information regarding miRNA regulation of TRAIL-mediated signaling in prostate cancer cells may provide potential biomarkers for the characterization of patients as responders and nonresponders of TRAIL-based therapy and could provide rationalized basis for combination therapies with TRAIL death receptor-targeting drugs.

Description: We report on the defect-dominated light emission and ultraviolet (UV) photoconductivity characteristics of ZnO nanorods (NRs) fabricated using a facile, cost-effective, and catalyst-free thermal decomposition route under varying reaction temperatures. The morphological and structural studies reveal the formation of homogeneous quality nanorods in large scale at the highest reaction temperature of 600  ∘ C. The luminescence feature of the nanorods is dominated by the defect related emission over the typical band edge emission. The variation of band-edge and native defect-related emission response of the samples has been correlated to the morphology and microstructure. In photoconductivity studies, the I – V characteristics of the ZnO NRs prepared at different reaction temperatures in dark and under UV illumination ( λ =365 nm) follow the power law, i.e., IαV r . An enhanced ultraviolet photodetection has been observed in the nanorods fabricated at the highest reaction temperature of 600  ∘ C. The sample prepared at highest reaction temperature of 600  ∘ C exhibits UV photosensitivity value (photo-to-dark current ratio) of around 1.18×10 3 , which is much higher in magnitude compared to that of the samples prepared at lower reaction temperatures. The enhanced photoconductivity may be assigned to the development of uniformity and homogeneity of the nanorods. Further development of such ZnO nanostructures can form the basis of promising prototype luminescent and UV photodetecting devices.

Description: The development of cellulase-based bioprocess is afflicted by the processing efficiency of enzymes. To address this issue, a method based on artificial oil bodies (AOBs) was proposed to integrate production and immobilization of recombinant cellulase. First, the heterologous endoglucanase ( celA ), cellobiohydrolase ( celK ), and β-glucosidase ( gls ) genes were individually fused with oleosin, a structural protein of plant seed oils. After expression in Escherichia coli , each fusion protein of insolubility was mixed together with plant oils. AOBs were assembled by subjecting the mixture to sonication. Consequently, active CelA, CelK, and Gls were resumed and co-immobilized on AOBs surface. Finally, the assembly condition (including the protein ratio) and the reaction condition were further optimized by response surface methodology. The resulting AOBs-bound cellulase remained stable for 4 cycles of cellulose–hydrolyzed reactions. Overall, the result shows a promise of this proposed approach for processing recombinant cellulase, which may provide a facile method to investigate optimum combination of cellulase components towards various cellulosic materials.

Description: Separation strategies based on size-selective precipitation of DNA fragments with polyethylene glycol (PEG) have been used for achieving desired DNA interval in automated sample preparation for next-generation sequencing. By varying PEG concentration, DNA fragments of different sizes can be precipitated onto surfaces of carboxyl-coated paramagnetic particles selectively, and therefore, the desired DNA interval can be obtained. However, one of the crucial points in this approach is to determine the critical PEG concentration for DNA fragment of a certain size. The aim of this work was to develop a convenient and reliable method for accurately determining the critical PEG concentration. In our method, at a fixed concentration of sodium chloride (NaCl), recovered DNA samples obtained with different PEG concentrations were directly quantified, and their concentrations as a function of the PEG concentration were fitted by the logistic function. The critical PEG value was easily and accurately determined from the fitted logistic function. The repeatability and stability of the critical PEG value were assessed, showing an excellent reliability of the method. Based on this method, critical PEG values of different-size DNA fragments were determined at different NaCl concentrations. The effectiveness of the method was also demonstrated by selective precipitation of DNA fragments.

Description: A recombinant oleate hydratase from Lysinibacillus fusiformis converted ricinoleic acid to a product, whose chemical structure was identified as the novel compound 10,12-dihydroxystearic acid by gas chromatograph/mass spectrometry, Fourier transform infrared, and nuclear magnetic resonance analysis. The reaction conditions for the production of 10,12-dihydroxystearic acid were optimized as follows: pH 6.5, 30 °C, 15 g l −1 ricinoleic acid, 9 mg ml −1 of enzyme, and 4 % ( v / v ) methanol. Under the optimized conditions, the enzyme produced 13.5 g l −1 10,12-dihydroxystearic acid without detectable byproducts in 3 h, with a conversion of substrate to product of 90 % ( w / w ) and a productivity of 4.5 g l −1  h −1 . The emulsifying activity of 10,12-dihydroxystearic acid was higher than that of oleic acid, ricinoleic acid, stearic acid, and 10-hydroxystearic acid, indicating that 10,12-dihydroxystearic acid can be used as a biosurfactant.

Description: Characteristics of metal–oxide–high- k –oxide–silicon (MOHOS) memories with oxygen-rich or oxygen-deficient GdO as charge storage layer annealed by NH 3 or N 2 are investigated. Transmission electron microscopy, X-ray photoelectron spectroscopy and X-ray diffraction are used to analyze the cross-sectional quality, composition and crystallinity, respectively, of the stacked gate dielectric with a structure of Al/Al 2 O 3 /GdO/SiO 2 /Si. The MOHOS capacitor with oxygen-rich GdO annealed in NH 3 exhibits a good trade-off among its memory properties: large memory window (4.8 V at ±12 V, 1 s), high programming speed (2.6 V at ±12 V/100 μs), good endurance and retention properties (window degradation of 5 % after 10 5 program/erase cycles and charge loss of 18.6 % at 85 °C after 10 years, respectively) due to passivation of oxygen vacancies, generation of deep-level traps in the grain boundaries of the GdO layer and suppression of the interlayer between GdO and SiO 2 by the NH 3 annealing.

Description: Tin targets immersed in ethanol and distilled water were ablated using a UV pulsed laser. The ablated products were investigated with transmission and scanning electron microscopy, X-ray photoelectron spectroscopy, and energy dispersive spectroscopy. For ablation in both liquids, the size distribution of the produced particles was bimodal, with particles having diameters of ∼10 nm and ∼1 μm. Formation mechanisms that caused the bimodal distribution are suggested. Ablation in ethanol resulted in nanoparticles that were found to be single crystals of tin coated with tin hydroxide (Sn(OH) 2 ) while ablation in water yielded nanoparticles that were polycrystalline tin dioxide (SnO 2 ) throughout.

Description: Thin aluminum oxide layers deposited on silicon by thermal atomic layer deposition can be used to reduce the electronic recombination losses by passivating the silicon surfaces. To activate the full passivation ability of such layers, a post-deposition annealing step at moderate temperatures (≈400  ∘ C, duration≈30 min) is required. Such an annealing step is commonly done in an oven in air, nitrogen, or forming gas atmosphere. In this work, we investigate the ability to reduce the duration of the annealing step by heating the silicon wafer with a microwave source. The annealing time is significantly reduced to durations below 1 min while achieving effective minority carrier lifetimes similar or higher to that of conventionally oven-annealed samples.

Description: A theoretical study is presented on the on/off current ratio limits for a ballistic coaxially-gated carbon nanotube field effect transistor (CNTFET) with highly doped source/drain regions. Based on changes in gate insulator dielectric constant and thickness, the current ratio has been estimated at different ambient temperatures. Decreasing the gate insulator thickness after a certain value around 3 nm causes the current ratio to degrade drastically. Although the higher dielectric constant values have a fair effect on current ratio, this effect could be suppressed when the device with a low gate insulator thickness works at a low ambient temperature. The simulation results also show that the temperature drastically degrades the current ratio value; whereas in a certain range of ambient temperature, tuning the values of gate insulator thickness and dielectric constant could be very helpful. In this way, the optimum values of gate insulator thickness and dielectric constant are identified to offer the highest on/off current ratio of the device.

Description: Bacillus amyloliquefaciens FZB42 has been shown to stimulate plant growth and to suppress the growth of plant pathogenic organisms including nematodes. However, the mechanism underlying its effect against nematodes remains unknown. In this study, we screened a random mutant library of B. amyloliquefaciens FZB42 generated by the mariner transposon Tn YLB-1 and identified a mutant strain F5 with attenuated nematicidal activity. Reversible polymerase chain reaction revealed that three candidate genes RAMB_007470 , yhdY , and prkA that were disrupted by the transposon in strain F5 potentially contributed to its decreased nematicidal activity. Bioassay of mutants impaired in the three candidate genes demonstrated that directed deletion of gene RBAM_007470 resulted in loss of nematicidal activity comparable with that of the F5 triple mutant. RBAM_007470 has been reported as being involved in biosynthesis of plantazolicin, a thiazole/oxazole-modified microcin with hitherto unknown function. Electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS) analyses of surface extracts revealed that plantazolicin bearing a molecular weight of 1,354 Da was present in wild-type B. amyloliquefaciens FZB42, but absent in the Δ RABM_007470 mutant. Furthermore, bioassay of the organic extract containing plantazolicin also showed a moderate nematicidal activity. We conclude that a novel gene RBAM_007470 and its related metabolite are involved in the antagonistic effect exerted by B. amyloliquefaciens FZB42 against nematodes.

Description: Pyramid-like spikes in a single crystal superalloy were investigated upon irradiation with picosecond (ps) laser pulses (200 ps, 800 nm, 1 kHz) under different laser fluences and pulse numbers. Both sides and grooves of pyramid-like spikes were covered with ripples, which had a period of ∼760 nm. The pyramid-like spike separation increased obviously with increasing laser fluence. Microstructural investigations indicate that the pyramid-like spikes were initiated with subsequent pulses from a smooth surface with corrugations and ripples. The coexistence of capillary waves for spikes and capillary waves for ripples in the melted material can be used to explain the formation of the pyramid-like spikes.

Description: This paper is to explore the effects of the liquid structure transition (LST) on the solidification kinetics of Sn-30 wt% Sb alloy by the Newton thermal analysis (NTA) method and the solidified microstructure analysis. Influence of the cooling rate on solidification behavior and microstructure was also concerned. With a self-designed sand mold, the cooling curves of five points were collected automatically in the process of solidification by a HYDRA. In the case of the liquid structure transition and a faster cooling rate, the modification melt treatment will lead to a higher undercooling of nucleation and an increased solidification latent heat in central part of solidifying castings, then the eventual grain size was evidently refined.

Description: Syngas fermentation is a promising route for resource recovery. Acetate is an important industrial chemical product and also an attractive precursor for liquid biofuels production. This study demonstrated high fraction acetate production from syngas (H 2 and CO 2 ) in a hollow-fiber membrane biofilm reactor, in which the hydrogen utilizing efficiency reached 100 % during the operational period. The maximum concentration of acetate in batch mode was 12.5 g/L, while the acetate concentration in continuous mode with a hydraulic retention time of 9 days was 3.6 ± 0.1 g/L. Since butyrate concentration was rather low and below 0.1 g/L, the acetate fraction was higher than 99 % in both batch and continuous modes. Microbial community analysis showed that the biofilm was dominated by Clostridium spp., such as Clostridium ljungdahlii and Clostridium drakei , the percentage of which was 70.5 %. This study demonstrates a potential technology for the in situ utilization of syngas and valuable chemical production.

Description: Lignosulfonates(LSs), by-products from chemical pulping processes, are low-value products with limited dispersion properties. The ability of commercially available horseradish peroxidase (HRP) to polymerize LS macromolecules and improve the dispersion properties of LSs was investigated. The polymerization of LSs proceeded efficiently under mild reaction conditions in an aqueous solution with HRP/H 2 O 2 . Gel permeation chromatography showed a significant increase in weight-average molecular weight ( M w ) of sulfonated kraft lignin and sodium lignosulfonate (NaLS) by 8.5-fold and 4.7-fold, respectively. The mechanism of polymerization was investigated by elemental analysis, surface charge measurement, headspace gas chromatography, infrared spectroscopy (IR), and hydrogen nuclear magnetic resonance spectrometry ( 1 H-NMR). The functional group measurements indicated that HRP incubation did not reduce the sulfonic group content. However, it decreased the phenolic and methoxyl group contents. As the phenolic group content decreased, M w increased as a power function. The polymerization was proposed to involve the random coupling of phenoxy radical intermediates. The radicals coupled with each other to form different inter-unit linkages, most of which were the β-O-4’ type, as the 1 H-NMR spectra indicated. Moreover, the HRP/H 2 O 2 incubation induced a significant improvement in the adsorption and dispersion properties of LSs. Therefore, the HRP/H 2 O 2 incubation is a promising approach for industrial applications of LSs.

Description: In the paper we report on picosecond-laser bulk microstructuring and stimulated Raman scattering (SRS) in type IIa single-crystal diamond in the course of multipulse irradiation at λ =532 nm wavelength using an advanced ps-laser system equipped with additional setups for on-line video imaging and photoluminescence spectra measurements. The effect of crystal orientation (relative to the incident laser beam) on (i) optical breakdown thresholds, (ii) character of bulk modifications, and (iii) generation of stimulated Raman scattering in diamond during irradiation with picosecond pulses of different durations ( τ 1 =10 ps and τ 2 =44 ps) is studied. It is shown that the processes of laser-induced breakdown in the bulk of diamond (at the backside of the crystals) and bulk microstructure growth are governed by the dielectric breakdown mechanism. It is found that generation of high-order stimulated Raman scattering in diamond crystals has a considerable effect on the threshold of laser-induced breakdown and bulk microstructuring. Conditions of the efficient SRS lasing are determined, depending on the pulse duration and the direction ([100] and [110]) of the laser beam incidence. A method of local temperature measurements in the bulk of diamond based on the Stokes-to-anti-Stokes intensity ratio in the recorded SRS spectra is proposed, its applicability to determine a “pre-breakdown” temperature of diamond during multipulse ps-laser irradiation is discussed.

Description: Marine microalga Nannochloropsis oculata possesses nutrients valuable for human health. In this study, we added freeze-dried N. oculata powder to soybean oil and observed a remarkable inhibition in oil oxidation. The amount of microalgae powder added was positively correlated to the increase in oil stability. The addition of 5.0 % ( w / w ) microalgae powder increased the oil stability index (OSI) values of soybean oil more than twofold at the tested temperatures 120 and 130 °C. N . oculata contains high levels of both phenolic compounds and α-tocopherols that could be the contributors to such an increase of the OSI. Two methods were conducted to assay the active ingredients released from microalgae: one employed three solvent systems to extract the microalgae and the other was the soybean oil added with microalgae. Analyses of free radical scavenging and reducing power suggested that the phenolic compounds dominated the antioxidation activities in soybean oil when it was infused with the microalgae powder. Our results suggest that N. oculata could potentially be used as an additive in cooking oil to increase the shelf life and nutritional value of the oil and to reduce the production of free radicals from lipid oxidation when the oil is used at high-temperature cooking processes.

Description: The actinomycete Streptomyces platensis produces two compounds that display antibacterial activity: platensimycin and platencin. These compounds were discovered by the Merck Research Laboratories, and a complex insoluble production medium was reported. We have used this medium as our starting point in our studies. In a previous study, we developed a semi-defined production medium, i.e., PM5. In the present studies, by varying the concentration of the components of PM5, we were able to develop a superior semi-defined medium, i.e., PM6, which contains a higher concentration of lactose. Versions of PM6, containing lower concentrations of all components, were also found to be superior to PM5. The new semi-defined production media contain dextrin, lactose, MOPS buffer, and ammonium sulfate in different concentrations. We determined antibiotic production capabilities using agar diffusion assays and chemical assays via thin-layer silica chromatography and high-performance liquid chromatography. We reduced crude nutrient carryover from the seed medium by washing the cells with distilled water. Using these semi-defined media, we determined that addition of the semi-defined component soluble starch stimulated antibiotic production and that it and dextrin could both be replaced with glucose, resulting in the chemically defined medium, PM7.

Description: Pichia pastoris is widely used as a host system for heterologous protein expression in both academia and industry. Production is typically accomplished by a fed-batch induction process that is known to have negative impacts on cell physiology that impose limits on both protein yields and quality. We have analysed recombinant protein production in chemostat cultures to understand the physiological responses associated with methanol-induced production of two human lysozyme variants with different degrees of misfolding by P . pastoris . Confounding variables associated with nutrient stress or growth-rate are minimised during steady-state growth in chemostats. Comparison of transcriptome-level data obtained during the non-inducing and inducing steady states identified changes in expression of only about 1 % of the genome during production of either an amyloidogenic human lysozyme variant prone to intracellular aggregation (I56T) or a misfolded but secretable variant (T70N), indicating near-complete acclimation to their production. A marked, but temporary, stress response involving both the unfolded protein response (UPR) and ER-associated degradation pathway was observed during the transient between steady states, particularly following induction of the T70N variant synthesis, and was accompanied by changes in expression of around 50 antisense transcripts. The results suggest that optimal heterologous protein production could best be achieved by a continuous process that minimises the number of methanol-induced transients experienced by the cultures. The processing of HAC1 mRNA required for the UPR was found to be constitutive in the culture conditions used, even in the absence of recombinant protein induction.

Description: The Aminobacter sp. strain MSH1 has potential for pesticide bioremediation because it degrades the herbicide metabolite 2,6-dichlorobenzamide (BAM). Production of the BAM-degrading bacterium using aerobic bioreactor fermentation was investigated. A mineral salt medium limited for carbon and with an element composition similar to the strain was generated. The optimal pH and temperature for strain growth were determined using shaker flasks and verified in bioreactors. Glucose, fructose, and glycerol were suitable carbon sources for MSH1 ( μ  = 0.1 h −1 ); slower growth was observed on succinate and acetic acid ( μ  = 0.01 h −1 ). Standard conditions for growth of the MSH1 strain were defined at pH 7 and 25 °C, with glucose as the carbon source. In bioreactors (1 and 5 L), the specific growth rate of MSH1 increased from μ  = 0.1 h −1 on traditional mineral salt medium to μ  = 0.18 h −1 on the optimized mineral salt medium. The biomass yield under standard conditions was 0.47 g dry weight biomass/g glucose consumed. An investigation of the catabolic capacity of MSH1 cells harvested in exponential and stationary growth phases showed a degradation activity per cell of about 3 × 10 −9  μg BAM h −1 . Thus, fast, efficient, large-scale production of herbicide-degrading Aminobacter was possible, bringing the use of this bacterium in bioaugmentation field remediation closer to reality.

Description: Copper oxide (CuO) is a p-type semiconductor with a band gap of 1.2 eV, which is well known in high-temperature superconductor and antiferromagnetic (AFM) materials through Cu–O–Cu super-exchange interaction. In this paper, we report the strong anisotropic ferromagnetism (FM) in aligned CuO nanorod arrays synthesized by a microwave-assisted hydrothermal method. The transmission electron microscopy (TEM) image shows that the CuO nanorod consists of a large number of smaller nanorods with almost the same growth direction. The X-ray diffraction (XRD) pattern indicates that the CuO nanorods are well crystallized with highly preferred orientation of the [020] direction. These CuO nanorod arrays show room-temperature ferromagnetism, with strong magnetic anisotropy when the magnetic field is applied perpendicular or parallel to the rod axis. This phenomenon of room-temperature ferromagnetism in those aligned CuO nanorods might originate from uncompensated surface spins and shape anisotropy of the nanorods.

Description: Soy sauce is a traditional condiment manufactured by natural inoculation and mixed culture fermentation. As is well known, it is the microbial community that plays an important role in the formation of its flavors. However, to date, its dynamic changes during the long period of fermentation process are still unclear, intensively constraining the improvement and control of the soy sauce quality. In this work, we revealed the dynamic changes of the microbial community by combining a cultured dependent method and a cultured independent method of polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis. Results indicated that the two methods verified and complemented each other in profiling microbial community, and that significant dynamics of the microbial community existed during the fermentation process, especially the strong inhibition of the growth of most of the microbes when entering into the mash stage from the koji stage. In the analysis of bacterial community, Staphylococcus and Bacillus were found to be the dominant bacteria and detected in the whole fermentation process. Kurthia and Klebsiella began to appear in the koji stage and then fade away in the early stage of the mash fermentation. In the analysis of fungal community, Aspergillus sojae and Zygosaccharomyces rouxii were found to be the dominant fungi in the koji and mash fermentation, respectively. It was clearly shown that when A. sojae decreased and disappeared in the middle stage of the mash fermentation, Z. rouxii appeared and increased at the meantime. Aspergillus parasiticus , Trichosporon ovoides and Trichosporon asahii also appeared in the koji and the early period of the mash fermentation and disappeared thereafter. Similar to Z. rouxii , Millerozyma farinosa and Peronospora farinosa were also found spontaneously which appeared in the mid-late period of the mash fermentation. The principal component analysis suggested that the microbial community underwent significant changes in the early period of the fermentation and, thereafter, tended to the stabilization in the mid-late periods. This study gave us important clues to understand the fermentation process and can serve as a foundation for improving the quality of soy sauce in the future.

Description: (TiVCrZrY)N coatings were deposited onto Si substrate by the radio-frequency (RF) magnetron sputtering of a TiVCrZrY alloy target in an N 2 /Ar atmosphere. The crystal, microstructural, mechanical, and electrical properties at different N 2 -to-total (N 2 +Ar) flow-rate ratio ( R N ) values were investigated. The coating produced in pure Ar had an equiaxed structure with a hexagonal-close-packed phase. With increased R N , the crystallinity and grain size markedly decreased. The microstructure of (TiVCrZrY)N coatings transformed from V-shaped columnar with a rough-domed surface into fine fibrous with a smooth surface. The amorphous transition layer above the substrate was also significantly thickened. The hardness of (TiVCrZrY)N decreased from 20.9 GPa to 18.9 GPa, and the electrical resistivity increased from 398.2 μΩ⋅cm to 21870 μΩ⋅cm.

Description: Galactosidases are widespread enzymes that are used for manifold applications, including production of prebiotics, biosynthesis of different transgalactosylated products, improving lactose tolerance and in various analytical approaches. The nature of these applications often require galactosidases to be present in a purified form with clearly defined properties, including precisely determined substrate specificities, low sensitivity to inhibitors, and high efficiency and stability under distinct conditions. In this study, we present the recombinant expression and purification of two previously uncharacterized β-galactosidases from Aspergillus nidulans as well as one β-galactosidase from Aspergillus niger . All enzymes were active toward p -nitrophenyl-β- d -galactopyranoside as substrate and displayed similar temperature and pH optima. The purified recombinant galactosidases digested various complex substrates containing terminal galactose β-1,4 linked to either N -acetylglucosamine or fucose, such as N -glycans derived from bovine fibrin and Caenorhabditis elegans . In our comparative study of the recombinant galactosidases with the commercially available galactosidase from Aspergillus oryzae , all enzymes also displayed various degrees of activity toward complex oligosaccharides containing β-1,3-linked terminal galactose residues. All recombinant enzymes were found to be robust in the presence of various organic solvents, temperature variations, and freeze/thaw cycles and were also tested for their ability to synthesize galactooligosaccharides. Furthermore, the use of fermentors considerably increased the yield of recombinant galactosidases. Taken together, we demonstrate that purified recombinant galactosidases from A. niger and from A. nidulans are suitable for various glycobiological and biotechnological applications.

Description: A Lolium perenne ice-binding protein ( Lp IBP) demonstrates superior ice recrystallization inhibition (IRI) activity and has proposed applications in cryopreservation, food texturing, as well as in being a “green” gas hydrate inhibitor. Recombinant production of Lp IBP has been previously conducted in bacterial and yeast systems for studies of protein characterization, but large-scale applications have been hitherto limited due to high production costs. In this work, a codon-optimized Lp IBP was recombinantly expressed and secreted in a novel one-step vector system from the nuclear genome of the green microalga Chlamydomonas reinhardtii . Both mixotrophic and photoautotrophic growth regimes supported Lp IBP expression, indicating the feasibility of low-cost production using minimal medium, carbon dioxide, and light energy as input. In addition, multiple growth and bioproduct extraction cycles were performed by repetitive batch cultivation trials, demonstrating the potential for semi-continuous production and biomass harvesting. Concentrations of recombinant protein reached in this proof of concept approach were sufficient to demonstrate IRI activity in culture media without additional purification or concentration, with activity further verified by thermal hysteresis and morphology assays. The incorporation of the recombinant Lp IBP into a model gas hydrate offers the promise that algal production may eventually find application as a “green” hydrate inhibitor.

Description: The interest in biofertilizers is increasing and so is the potential for their use in sustainable agriculture. However, many of the products that are currently available worldwide are often of very poor quality, resulting in the loss of confidence from farmers. The formulation of an inoculant is a crucial multistep process that should result in one or several strains of microorganisms included in a suitable carrier, providing a safe environment to protect them from the often harsh conditions during storage and ensuring survival and establishment after introduction into soils. One of the key issues in formulation development and production is the quality control of the products, at each stage of the process. This review presents the different components and the major steps involved in the formulation of good quality biofertilizers, including the techniques used to assess the quality of the products following production. The quality of currently available inoculants is also reviewed, emphasizing the need for better quality control systems worldwide.

Description: A large number of molds serve as producer strains for the industrial production of pharmaceuticals, foods, or organic chemicals. To optimize strains for production processes, conventional strain development programs use random mutagenesis and, more recently, recombinant technologies to generate microbial strains with novel and advantageous properties. The recent detection of mating type genes in fungal production strains and the discovery of cryptic sexuality in presumably asexual fungi open up novel strategies for generating progeny with new, as yet unobserved properties. Mating type genes, which can be considered as “sex genes,” not only direct sexual development but also regulate a broad range of fungal secondary metabolites. In addition, they control hyphal morphology, which has a direct impact on production processes that are often conducted in huge fermenter tanks. Here, we survey the occurrence and function of mating type genes that have been discovered in a wide range of industrial fungal producer strains. The possibility to obtain progeny from industrial producers by sexual mating provides an exciting alternative to conventional strain improvement programs aiming to generate optimized recombinant production strains.

Description: Endo-1,4-β-xylanases (EC 3.2.1.8) hydrolyze the 1,4-β-D-xylosidic linkages in xylans, the most abundant hemicellulose in plant cell walls. Xylanase enzymes have numerous industrial applications, including the manufacturing of animal feed, bread, juice and wine, pulp and paper, and biofuels. In this study, two glycosyl hydrolase family 10 members designated Gt Xyn10A and Gt Xyn10B and two glycosyl hydrolase family 11 members, Op Xyn11A and Cc Xyn11C, were functionally expressed and subjected to biochemical characterization. The K M , V max , and k cat values of the four xylanases, determined using birchwood xylan, ranged from 0.27 to 1.1 mg/mL, 130 to 980 μmol/min/mg, and 109 to 344 s −1 , respectively, where Op Xyn11A gave the highest and Gt Xyn10B the lowest values for all three parameters. Substrate specificity studies and analysis of the products released during the degradation of xylo-oligosaccharides and three types of xylan revealed significant differences in catalytic properties, particularly between Op Xyn11A and the other xylanases and between the family 10 and the family 11 xylanases. Molecular modeling suggests that the unique substrate specificity of Op Xyn11A can be attributed to the presence of a serine rather that an asparagine or aspartate residue at the +1 substrate binding site. Additionally, all four xylanases exhibited biochemical characteristics of interest for various commercial applications.

Description: Hand, foot, and mouth disease (HFMD) has caused significant morbidity and mortality in the Asia-Pacific regions, particularly in infants and young children. Coxsackievirus A16 (CA16) represents one of the major causative agents for HFMD, and the development of a safe and effective vaccine preventing CA16 infections has become a public health priority. In this study, we have developed a yeast system for the production of virus-like particles (VLPs) for CA16 by co-expressing P1 and 3CD of CA16 in Saccharomyces cerevisiae . These VLPs exhibit similarity in both protein composition and morphology as empty particles from CA16-infected cells. Immunization with CA16 VLPs in mice potently induced CA16-specific IgG and neutralization antibodies in a dose-dependent manner. IgG subclass isotyping revealed that IgG1 and lgG2b were dominantly induced by VLPs. Meanwhile, cytokine profiling demonstrated that immunization with VLPs significantly induced the secretion of IFN-γ, indicating potent cellular immune response. Furthermore, in vivo challenge experiments showed that passive immunization with anti-VLPs sera conferred full protection against lethal CA16 challenge in neonate mice. Taken together, our data demonstrated that VLPs produced in yeast might have the potential to be further developed as a vaccine candidate against HFMD.

Description: Studies were carried out to assess the utility of the cellular and extracellular constituents of Bacillus megaterium for the flotation of sphalerite and galena minerals. Based on the flotation results on the individual minerals, it was observed that sphalerite was preferentially floated compared to galena. A maximum selectivity index (SI) value of 11.7 was achieved in the presence of the soluble fraction of the thermolysed cells, which was higher than that obtained with the intact cells (SI of 6.5) and the insoluble fraction of the thermolysed cells (SI of 9.6). The results of the various enzymatic treatment tests revealed that extracellular DNA played a vital role in the selective flotation of sphalerite. A noteworthy finding was that the single-stranded DNA (ssDNA) had a higher biocollector capacity vis-à-vis the double-stranded DNA (dsDNA), leading to better flotation efficiency. About 95 % recovery of sphalerite could be achieved from the mineral mixture by the combined addition of the ssDNA with the non-DNA components of the bacterial cells, resulting in a maximum SI of 19.1. Calcium and phosphate components of the nutrient media were found to be essential for better selectivity of separation of sphalerite. The mechanisms of microbe–mineral interaction are discussed.

Description: While the use of anaerobic digestion to generate methane as a source of bioenergy is increasing worldwide, our knowledge of the microbial communities that perform biomethanation is very limited. Using next-generation sequencing, bacterial population profiles were determined in three full-scale mesophilic anaerobic digesters operated on dairy farms in the state of Vermont (USA). To our knowledge, this is the first report of a metagenomic analysis on the bacterial population of anaerobic digesters using dairy manure as their main substrate. A total of 20,366 non-chimeric sequence reads, covering the V1-V2 hypervariable regions of the bacterial 16S rRNA gene, were assigned to 2,176 operational taxonomic units (OTUs) at a genetic distance cutoff value of 5 %. Based on their limited sequence identity to validly characterized species, the majority of OTUs identified in our study likely represented novel bacterial species. Using a naïve Bayesian classifier, 1,624 anaerobic digester OTUs could be assigned to 16 bacterial phyla, while 552 OTUs could not be classified and may belong to novel bacterial taxonomic groups that have yet to be described. Firmicutes , Bacteroidetes , and Chloroflexi were the most highly represented bacteria overall, with Bacteroidetes and Chloroflexi showing the least and the most variation in abundance between digesters, respectively. All digesters shared 132 OTUs, which as a “core” group represented 65.4 to 70.6 % of sequences in individual digesters. Our results show that bacterial populations from microbial communities of anaerobic manure digesters can display high levels of diversity despite sharing a common core substrate.

Description: The discovery of Dehalococcoides mccartyi reducing perchloroethene and trichloroethene (TCE) to ethene was a key landmark for bioremediation applications at contaminated sites. D. mccartyi -containing cultures are typically grown in batch-fed reactors. On the other hand, continuous cultivation of these microorganisms has been described only at long hydraulic retention times (HRTs). We report the cultivation of a representative D. mccartyi -containing culture in continuous stirred-tank reactors (CSTRs) at a short, 3-d HRT, using TCE as the electron acceptor. We successfully operated 3-d HRT CSTRs for up to 120 days and observed sustained dechlorination of TCE at influent concentrations of 1 and 2 mM TCE to ≥97 % ethene, coupled to the production of 10 12 D. mccartyi cells L culture −1 . These outcomes were possible in part by using a medium with low bicarbonate concentrations (5 mM) to minimize the excessive proliferation of microorganisms that use bicarbonate as an electron acceptor and compete with D. mccartyi for H 2 . The maximum conversion rates for the CSTR-produced culture were 0.13 ± 0.016, 0.06 ± 0.018, and 0.02 ± 0.007 mmol Cl − L culture −1 h −1 , respectively, for TCE, cis -dichloroethene, and vinyl chloride. The CSTR operation described here provides the fastest laboratory cultivation rate of high-cell density Dehalococcoides cultures reported in the literature to date. This cultivation method provides a fundamental scientific platform for potential future operations of such a system at larger scales.

Description: The blue InGaN light-emitting diodes (LEDs), employing a lattice-compensated p-AlGaN/InGaN superlattice (SL) interlayer to link the last quantum barrier and electron blocking layer (EBL), are proposed and investigated numerically. The simulation results indicate that the newly designed LEDs have better hole injection efficiency, lower electron leakage, and smaller electrostatic fields in the active region over the conventional LEDs mainly attributed to the mitigated polarization-induced downward band bending. Furthermore, the markedly improved output power and efficiency droop are also suggested when the conventional LEDs corresponding to experiment data are replaced by the newly designed LEDs.

Description: In this paper, we investigate the resonance magnetoelectric (ME) effect in the middle supported multilayer composites consisting of high-permeability Fe-based nanocrystalline soft magnetic alloy Fe 73.5 Cu 1 Nb 3 Si 13.5 B 9 (FeCuNbSiB), Nickel (Ni), and piezoelectric Pb(Zr 1− x Ti x )O 3 (PZT). The coupling effect between positive magnetostrictive FeCuNbSiB and negative magnetostrictive Ni results in the build-in magnetic bias due to their different magnetic permeability and coercivity. As a result, a giant resonance ME voltage coefficient ( α ME, r ) at zero DC magnetic bias field ( H dc ) and multi-peaks of α ME, r for FeCuNbSiB/Ni/PZT/Ni/FeCuNbSiB composite are observed. The experimental results show that the giant zero-biased α ME, r strongly depends on the thickness of FeCuNbSiB ribbon. The maximum zero-biased α ME, r is up to 86 V/cm Oe for FeCuNbSiB/Ni/PZT/Ni/FeCuNbSiB with four-layer FeCuNbSiB ribbons, which is ∼500 times higher than that of the previously reported NKNLS-NZF/Ni/NKNLS-NZF trilayer composite. Compared with the peak α ME, r and the optimum H dc of Ni/PZT/Ni composite, the largest peak α ME, r of FeCuNbSiB/Ni/PZT/Ni/FeCuNbSiB composite with four-layer FeCuNbSiB ribbons increases ∼185 %, and the optimum H dc decreases ∼300 Oe, respectively. Based on the nonlinear magnetostrictive constitutive relation and the magnetoelectric equivalent circuit, a theoretical model of α ME, r versus H dc is built under free boundary conditions. Calculated zero-biased α ME, r and α ME, r versus H dc are in good agreement with the experimental data. This laminate composite shows promising applications for high-sensitivity power-free magnetic field sensors, zero-biased ME transducers and small-size energy harvesters.

Description: In this paper, we report on in-situ atomic force microscopy (AFM) studies of topographical changes in azobenzene-containing photosensitive polymer films that are irradiated with light interference patterns. We have developed an experimental setup consisting of an AFM combined with two-beam interferometry that permits us to switch between different polarization states of the two interfering beams while scanning the illuminated area of the polymer film, acquiring corresponding changes in topography in-situ . This way, we are able to analyze how the change in topography is related to the variation of the electrical field vector within the interference pattern. It is for the first time that with a rather simple experimental approach a rigorous assignment can be achieved. By performing in-situ measurements we found that for a certain polarization combination of two interfering beams [namely for the SP (↕, ↔) polarization pattern] the topography forms surface relief grating with only half the period of the interference patterns. Exploiting this phenomenon we are able to fabricate surface relief structures with characteristic features measuring only 140 nm, by using far field optics with a wavelength of 491 nm. We believe that this relatively simple method could be extremely valuable to, for instance, produce structural features below the diffraction limit at high-throughput, and this could significantly contribute to the search of new fabrication strategies in electronics and photonics industry.

Description: Traditional glass micromachining using laser processing in air would produce many kinds of defects, such as bulges, debris, micro-cracks and scorches. In this article, a poly-dimethylsiloxane (PDMS) protection processing has been presented to reduce the temperature gradient and heat-affected zone (HAZ) to achieve crack-free Pyrex glass machining. A good quality of etched surface which is a clear and much-reduced bulge without crack and scorch is achieved using CO 2 laser micromachining at 150 μm thick PDMS protection layer and the laser powers of 10–15 W and scanning speeds of 228–342 mm/s for five passes. The PDMS cover layer benefits feature size and bulge height reduction. The alpha-step measured profile shows that the much reduced bulge height around the rims of channel was about 1.2 μm at 150 μm thick PDMS about 13 times smaller than that in air. The ANSYS software was used to analyze the temperature distribution and thermal stress field of glass micromachining in air without and with PDMS cover layer. The smaller temperature gradient observed in PDMS protection processing has the smaller HAZ and diminishes the crack formation during the laser processing.

Description: To reduce the cost of the emitter diffusion process, there has been increasing interest to substitute the standard process of batch POCl 3 emitter diffusion used in the silicon solar-cell manufacturing industry with in-line diffusion processes such as the spray-on and screen-printing process. For this reason, it is essential to study and compare the processes of different diffusion methods from the point of view of the crystalline quality of the final wafers. X-ray transmission topography was employed to characterize the possible precipitates and other microdefects generated in Czochralski-grown silicon (Cz Si) during the emitter diffusion process carried out by screen-printing, spray-on and the standard process, in which the emitter was provided by a liquid (POCl 3 ) source. The results indicate that the phosphorus diffusion process influences the crystalline quality of the wafers and the efficiency of the external gettering process that takes place during phosphorus diffusion depends on the diffusion method employed.

Description: We propose the use of a La 2 O 3 (LO) film as the capping layer for improvement of a semiconductor/insulator interface in a solution-processed indium–tin–oxide (ITO) ferroelectric-gate thin-film transistor (FGT) device. It is demonstrated that the LO layer acts as a good barrier film not only for preventing the interdiffusion between the ITO semiconductor and lead–zirconium-titanate (PZT) insulator layers, but also for stabilizing the PZT surface structure. The fabricated FGT device exhibited high I on / I off , large M w , high μ FE and improved retention time of about 10 9 , 3.5 V, 7.94 cm 2  V −1  s −1 and 1 day, respectively, which are comparable to or better than those obtained with FGTs fabricated by means of conventional vacuum processes. We also point out that the key origin of the interface improvement is likely due to the incorporation of La into the PZT system, forming a La surface-modified PZT system which is more stable than the pure PZT in terms of Pb volatility and formation of oxygen vacancies.

Description: The results of ferroelectric properties studies of KNN doped with La and Ti and sintered at temperatures in the interval of 1100 °C–1190 °C are presented in this work. The doping was achieved by the substitution of La for ions in A sites and Ti for ions in the B sites. Values of 94 % of the theoretical density were accomplished. The effect of the sintering temperature and the inclusion of the La and Ti cations in the KNN structure is evident through the shift in the ferroelectric-paraelectric transition temperature of ∼110 °C with respect to that of pure KNN (420 °C). Microstructure and ferroelectric analyses were carried out using Piezoresponse Force Microscopy (PFM) and hysteresis loops with interesting results, Δ P r =9 (μC/cm 2 ) and P r / P max =0.41, even when the saturation of the materials is not reached.

Description: We have developed a high-performance laser energy meter based on anisotropic Seebeck effect in a strongly correlated electronic (SCE) thin film. SCE thin films, typically represented by high-temperature superconductor (HTS) cuprate and colossal magnetoresistance (CMR) manganite thin films, demonstrate tremendous anisotropic Seebeck effect. In this study, a La 2/3 Ca 1/3 MnO 3 thin film grown on a tilted LaAlO 3 substrate is tested with the fundamental, the second, the third, and the fourth harmonics (1064, 532, 355, 266 nm, respectively) of a Q-switched Nd:YAG laser over a wide range of temperatures from room temperature to 16 K. The peak-value of the laser-induced thermoelectric voltage signal shows a good linear relationship with the laser energy per pulse in the measured wavelength and temperature ranges. The combined advantages over other commercial laser detectors such as nanosecond-order response and spectrally broad and flat response over a wide range of temperatures, in situ real-time measurement, and energy savings, make the device an ideal candidate for next-generation laser detectors and laser power/energy meters.

Description: Co-doped ZnO epilayer films were grown by pulsed laser deposition (PLD) on vicinal cut silicon and sapphire substrates. Changes in deposition time were observed as a moderate effect on the quality of the films, and the influence of the thickness on thermoelectric signals from Zn 0.9 Co 0.1 O thin films were discussed. The effect of one of the main deposition parameters, the deposition time, on the crystallinity and electron mobility properties of the Zn 0.9 Co 0.1 O thin films grown on sapphire was investigated by means of X-ray diffraction (XRD) and laser-induced voltage (LIV) effect. It shown that the XRD rocking curve full-width half-maximun (FWHM) decreased as time increasing, and the LIV signals were observed along the tilting angle of the substrate orientation when the pulsed KrF excimer laser of 248 nm were irradiated on the films. When the films illuminated in pulse lasers, the highest signals occurred in the films with best crystalline quality, and the signals were higher in the films grown on sapphire than those on silicon substrates. It suggested that the electrical resistivity and electron mobility have close relations with not only the crystallinity but also with the interface of the thin films.

Description: Nanostructure formation on bulk noble metals (copper, gold and silver) by a femtosecond laser was studied aiming at the production of low-reflectivity surfaces. The target surface was irradiated with the beam of a 775 nm wavelength and 150 fs pulse duration Ti:sapphire laser. The fluence was in the 16–2000 mJ/cm 2 range, while the average pulse number was varied between 10 and 1000 depending on the scanning speed of the sample stage. The reflectivity of the treated surfaces was measured with a visible–near-infrared microspectrometer in the 450–800 nm range, while the morphology was studied with a scanning electron microscope. A strong correlation was found between the decreasing reflectivity and the nanostructure formation on the irradiated surface; however, the morphology of silver significantly differed from those of copper and gold. For the two latter metals a dense coral-like structure was found probably as a result of cluster condensation in the ablation plume followed by diffusion-limited aggregation. In the case of silver the surface was covered by nanodroplets, which formation was probably influenced by the ‘spitting’ caused by ambient oxygen absorption in the molten silver followed by its fast release during the resolidification.

Description: The automation in fabrication of CFRP (carbon-fiber-reinforced plastics) parts demands efficient and low-cost machining technologies. In conventional cutting technologies, tool-wear and low process speeds are some of the reasons for high costs. Thus, the use of lasers is an attractive option for cutting CF-preforms. A typical effect degrading the quality in laser cutting CF-preform is a bulged cutting edge. This effect is assumed to be caused by interaction of the fibers with the ablated material, which leaves the kerf at high velocity. Hence, a method for measuring the momentum and the velocity of the vapor is presented in this article. To measure the momentum of the ablated material, the CF-preform is mounted on a precision scale while cutting it with a laser. The direction of the momentum was determined by measuring the momentum parallel and orthogonal to the CF-preform surface. A change of the direction of the momentum with different cutting-speeds is assessed at constant laser-power. Averaged velocities of the ablation products of up to 300 m/s were determined by measuring the ablated mass and the momentum.

Description: κ-Carrageenases exhibit apparent distinctions in gene sequence, molecular weight, enzyme properties, and posttranslational processes. In this study, a new κ-carrageenase gene named cgk Z was cloned from the marine bacterium Zobellia sp. ZM-2. The gene comprised an open reading frame of 1,638 bp and encoded 545 amino acids. The natural signal peptide of κ-carrageenase was used successfully for the secretory production of the recombinant enzyme in Escherichia coli . A posttranslational process that removes an amino acid sequence of about 20 kDa from the C-terminal end of κ-carrageenase was first discovered in E. coli . An increase in enzyme activity by 167.3 % in the presence of 5 mM DTT was discovered, and Na + at a certain concentration range was positively correlated with enzyme activity. The κ-carrageenase production of E. coli was 9.0 times higher than that of ZM-2. These results indicate the potential use of the enzyme in the biotechnological industry.

Description: Fifty-five bacterial isolates were obtained from surface-sterilized nodules of woody and shrub legumes growing in Ethiopia: Crotalaria spp., Indigofera spp., and Erythrina brucei , and the food legumes soybean and common bean. Based on partial 16S rRNA gene sequence analysis, the majority of the isolates were identified as Gram-negative bacteria belonging to the genera Achromobacter , Agrobacterium , Burkholderia , Cronobacter , Enterobacter , Mesorhizobium , Novosphingobium , Pantoea , Pseudomonas , Rahnella , Rhizobium , Serratia , and Variovorax . Seven isolates were Gram-positive bacteria belonging to the genera Bacillus , Paenibacillus , Planomicrobium , and Rhodococcus . Amplified fragment length polymorphism (AFLP) fingerprinting showed that each strain was genetically distinct. According to phylogenetic analysis of recA , glnII , rpoB , and 16S rRNA gene sequences, Rhizobium , Mesorhizobium , and Agrobacterium were further classified into six different genospecies: Agrobacterium spp., Agrobacterium radiobacter , Rhizobium sp., Rhizobium phaseoli , Mesorhizobium sp., and putative new Rhizobium species. The strains from R . phaseoli , Rhizobium sp. IAR30, and Mesorhizobium sp. ERR6 induced nodules on their host plants. The other strains did not form nodules on their original host. Nine endophytic bacterial strains representing seven genera, Agrobacterium , Burkholderia , Paenibacillus , Pantoea , Pseudomonas , Rhizobium , and Serratia , were found to colonize nodules of Crotalaria incana and common bean on co-inoculation with symbiotic rhizobia. Four endophytic Rhizobium and two Agrobacterium strains had identical nifH gene sequences with symbiotic Rhizobium strains, suggesting horizontal gene transfer. Most symbiotic and nonsymbiotic endophytic bacteria showed plant growth-promoting properties in vitro, which indicate their potential role in the promotion of plant growth when colonizing plant roots and the rhizosphere.

Description: Escherichia coli BA002, in which the ldhA and pflB genes are deleted, cannot utilize glucose anaerobically due to the inability to regenerate NAD + . To restore glucose utilization, overexpression of nicotinic acid phosphoribosyltransferase (NAPRTase) encoded by the pncB gene, a rate-limiting enzyme of NAD(H) synthesis pathway, resulted in a significant increase in cell mass and succinate production under anaerobic conditions. However, a high concentration of pyruvate accumulated. Thus, co-expression of NAPRTase and the heterologous pyruvate carboxylase (PYC) of Lactococcus lactis subsp. cremoris NZ9000 in recombinant E . coli BA016 was investigated. The total concentration of NAD(H) was 9.8-fold higher in BA016 than in BA002, and the NADH/NAD + ratio decreased from 0.60 to 0.04. Under anaerobic conditions, BA016 consumed 17.50 g l −1 glucose and produced 14.08 g l −1 succinate with a small quantity of pyruvate. Furthermore, when the reducing agent dithiothreitol or reduced carbon source sorbitol was added, the cell growth and carbon source consumption rate of BA016 was reasonably enhanced and succinate productivity increased.

Description: We report a design method of surface plasmon polaritons sharp bends based on transformation optics. Plasmonic waveguide bends with different angles, which possess little radiation loss, are proposed. Transformation media can be simply achieved with homogeneous and nonmagnetic materials, which can be constructed by altering two different dielectric films. Electromagnetic simulations by a finite-element method on detailed examples have been performed to validate the designs.

Description: This article presents the results on the growth and characterization of BaSnF 4 thin films on glass substrates prepared by pulsed laser deposition technique. The structural results of BaSnF 4 thin film carried out by glancing angle X-ray diffraction technique indicates the formation of the film with similar structure (tetragonal, P 4 /nmm) to the bulk target material. The absorption coefficient and band gap of the film is determined by suitable analysis of the transmittance spectra. The transport properties of the thin films are studied using impedance spectroscopy in the temperature range of 323–573 K. The frequency-dependent imaginary part of impedance plot shows that the conductivity relaxation is non-Debye in nature. The scaling behavior of the imaginary part of impedance at various frequencies indicates temperature-independent relaxation behavior.

Description: 1,2-Benzenedicarboxaldehyde-3,4,5-trihydroxy-6-methyl (flavipin) was found to be antagonistic against nematodes and fungi. Here we demonstrated that flavipin is a potent antioxidant in vitro and in vivo, which has great potential in the therapy for free radical-associated diseases. Therefore, flavipin-producing bio-source was screened from 80 endophytes in Ginkgo biloba . Seven endophytic fungi were able to synthesize antioxidant substances and identified by ITS rDNA sequences. Among them, Chaetomium globosum CDW7 was a remarkable producer of flavipin. The fermentation parameters of CDW7 were then optimized for high flavipin production. Cultured under the optimal condition (25 °C, 100/250 mL flask, 12 discs/flask, 150 rpm, pH 6.5) for 14 days, CDW7 was able to synthesize flavipin at a production of 315.5 mg/L. In addition, flavipin output was positively correlated to antioxidant activities of crude extracts with a correlation coefficient of 0.8235, indicating that flavipin was the major antioxidant component of CDW7's metabolites. These data demonstrated that CDW7 was a highly yielded bio-source of antioxidant flavipin.

Description: UV irradiation of materials consisting of a polymer matrix that possesses precursors of noble metals followed by annealing results in creation of metal nanoparticles within the irradiated domains. Such photoinduced nanocomposites are promising for photonics applications due to the strong alteration of their optical properties compared to initial nonirradiated materials. We report our results on the synthesis and investigation of two kinds of these materials: Photoinduced Au nanocomposites based on PMMA matrices, including bulk materials prepared by means of the polymerization technique; photoinduced Ag nanocomposites with an organic–inorganic hybrid matrix based on TiO 2 gels. The experimental data on evolution of absorption spectra of these materials due to laser irradiation at different wavelengths are presented. The linear and nonlinear refractive index changes in these materials owing to light-induced nanonstructuring are investigated.

Description: The study of pollution performance on a wind turbine blade due to lightning is important, as it can cause major damage to wind turbine blades. In the present work, optical emission spectroscopy (OES) technique is used to understand the influence of pollutant deposited on a wind turbine blade in an off-shore environment. A methodical experimental study was carried out by adopting IEC 60507 standards, and it was observed that the lightning discharge propagates at the interface between the pollutant and the glass fiber reinforced plastic (Material used in manufacturing of wind turbine blades). In addition, as a diagnostic condition monitoring technique, laser-induced breakdown spectroscopy (LIBS) is proposed and demonstrated to rank the severity of pollutant on the wind turbine blades from a remote area. Optical emission spectra observed during surface discharge process induced by lightning impulse voltage is in agreement with the spectra observed during LIBS.

Description: We developed a method that accurately determines an unknown position of the high-intensity laser-pulse-material interaction site on the front side of a plate. It is based on interferometric measurements of a normal displacement at known positions on the plate’s rear side. The displacement is caused by reflections of various pulsed-laser-induced mechanical waves. We have superseded the long-established time-of-flight approach with the improved, triple-echo method. To accurately locate the origin of the laser-induced ultrasound on the plate with a known thickness, we only need to detect the arrivals of the first three consecutive mode unconverted waves. Our method works without knowing the propagation velocities of various ultrasonic waves and additionally solves some time-related drawbacks of the conventional time-of-flight approach. The relative uncertainty of the measured source-receiver separations obtained with the presented method is less than 0.01.

Description: Laser dry etching by a laser driven direct writing apparatus has been extensively used for the micro- and nano-patterning on the solid surface. The purpose of this study is to pattern the PEDOT:PSS thin film coated on the soda-lime glass substrates by a nano-second pulsed ultraviolet laser processing system. The patterned PEDOT:PSS film structure provides the electrical isolation and prevents the electrical contact from each region for capacitive touch screens. The surface morphology, geometric dimension, and edge quality of ablated area after the variety of laser patternings were measured by a 3D confocal laser scanning microscope. After the single pulse laser irradiation, the ablation threshold of the PEDOT:PSS film conducted by the nano-second pulsed UV laser was determined to be 0.135±0.003 J/cm 2 . The single pulse laser interacted region and the ablated line depth increased with increasing the laser fluence. Moreover, the inner line width of ablated PEDOT:PSS films along the patterned line path increased with increasing the laser fluence but the shoulder width increased with decreasing fluence, respectively. The clean, smooth, and straight ablated edges were accomplished after the electrode patterning with the laser fluence of 1.7 J/cm 2 and 90 % overlapping rate.

Description: Selectiveness of the laser processing is the top-most important for applications of the processing technology in thin-film electronics, including photovoltaics. Coupling of laser energy in multilayered thin-film structures, depending on photo-physical properties of the layers and laser wavelength was investigated experimentally and theoretically. Energy coupling within thin films highly depends on the film structure. The finite element and two-temperature models were applied to simulate the energy and temperature distributions inside the stack of different layers of a thin-film solar cell during a picosecond laser irradiation. Reaction of the films to the laser irradiation was conditioned by optical properties of the layers at the wavelength of laser radiation. Simulation results are consistent with the experimental data achieved in laser scribing of copper-indium-gallium diselenide (CIGS) solar cells on a flexible polymer substrate using picosecond-pulsed lasers. Selection of the right laser wavelength (1064 nm or 1572 nm) enabled keeping the energy coupling in a well-defined volume at the interlayer interface. High absorption at inner interface of the layers triggered localized temperature increase. Transient stress caused by the rapid temperature rise facilitating peeling of the films rather than evaporation. Ultra-short pulses ensured high energy input rate into absorbing material permitting peeling of the layers with no influence on the remaining material.

Description: Two bacterial hosts expressing cloned aromatic oxygenases were used to catalyze the oxidation and polymerization of indole and related substrates, creating mixtures of indigoid compounds comprised of novel dimers and trimers. Crude extracts and purified compounds were tested for their ability to inhibit the growth of Gram-positive organisms, in general, and Mycobacterium tuberculosis (TB), in particular. Of the 74 compounds tested against M. tuberculosis , ~66 % had minimum inhibitory concentrations (MIC) of 5 μg/ml or less. The most effective antibiotic found was designated SAB-P1, a heterodimer of indole and anthranil, which had a MIC of 0.16 μg/ml, and did not inhibit kidney cells (IC 50 ) at concentrations of 〉8 μg/ml. Combinatorial biocatalysis was used to create a series of halogenated derivatives of SAB-P1 with a wider therapeutic window. None of the derivatives had MIC values that were superior to SAB-P1, but some had a wider therapeutic window because of decreased kidney cell toxicity. Generally, the indigoid dimers that were effective against TB appeared to be specific for TB. Some of the trimers generated, however, had a broader spectrum of activity inhibiting not only TB (MIC = 1.1 μg/ml) but also the growth of Mycobacterium smegmatis MC2 155, Bacillus cereus , Enterococcus faecalis , Staphylococcus epidermidis , Bacillus subtilis 168, and Clostridium acetobutylicum . The structure of two of the novel dimers (SAB-C4 and SAB-P1) and a trimer (SAB-R1) were solved using X-ray crystallography.

Description: The compartmentalized anaerobic reactor (CAR) is a patent novel high-rate reactor, which consists of three compartments. The reactor has a great potential for application due to its many advantages. In this work, the microbial consortium, spatial distribution, and their relationship with performance of CAR were investigated by means of polymerase chain reaction, denaturing gradient gel electrophoresis, and fluorescence in situ hybridization. The results showed that the predominant archaea were Methanobacterium , Methanosaeta , and Methanospirillum , and the predominant bacteria were Firmicutes , Deltaproteobacteria , Spirochaetes , Actinobacteria , and Gammaproteobacteria in the microbial consortium. The methanogenic archaea (MA), the hydrogen-producing acetogenic bacteria (HAB), and the hydrolytic fermentative bacteria (HFB) were found to be predominant in the upper, middle, and bottom compartments, respectively. The results revealed that the granular sludge took on a stratified microbial structure. The HFB, HAB, and MA were located in the outer shell, middle layer, and core, respectively. The microbial populations from the bottom compartment were relatively homogeneous in the granular sludge, and from the middle and upper compartments, they were relatively heterogeneous in the granular sludge. The microbial consortia and their spatial distribution were in accordance with the organic loading rate and chemical components in the three compartments.

Description: We constructed beta-glucosidase (BGL)-displaying Corynebacterium glutamicum , and direct l -lysine fermentation from cellobiose was demonstrated. After screening active BGLs, Sde1394, which is a BGL from Saccharophagus degradans , was successfully displayed on the C. glutamicum cell surface using porin as an anchor protein, and cellobiose was directly assimilated as a carbon source. The optical density at 600 nm of BGL-displaying C. glutamicum grown on cellobiose as a carbon source reached 23.5 after 48 h of cultivation, which was almost the same as that of glucose after 24 h of cultivation. Finally, Sde1394-displaying C. glutamicum produced 1.08 g/l of l -lysine from 20 g/l of cellobiose after 4 days of cultivation, which was about threefold higher than the amount of produced l -lysine using BGL-secretory C. glutamicum strains (0.38 g/l after 5 days of cultivation). This is the first report on amino acid production using cellobiose as a carbon source by BGL-expressing C. glutamicum .

Description: An integrated microchemical–petrographic approach is here proposed to discriminate the provenance of archaeological pottery artefacts from distinct production centres. Our study focuses on a statistically significant sampling ( n =186) of volcanic temper-bearing potteries representative of the manufacturing and dispersion among the islands of the Aeolian Archipelago during the Bronze Age. The widespread establishment of new settlements and the abundant recovery of Aeolian-made ceramic in southern Italy attest for the increased vitality of the Archipelago during the Capo Graziano culture (Early Bronze Age–Middle Bronze Age 2; 2300–1430 BC). Potteries from three of the main known ancient communities (Lipari, Filicudi and Stromboli) have been studied integrating old collections and newly excavated material. Volcanic tempers have been first investigated through multivariate analyses of relative abundances of mineral and rock clasts along with petrographic characters. In addition, we performed in-situ mineral chemistry microanalyses by Electron Microprobe and Laser Ablation—Inductively Coupled Plasma Mass Spectrometry to assess major and trace element composition of the most common mineral phases. Four Temper Compositional Reference Units have been recognised based on compositional trends. Two units (AI and AX) are unequivocally distinct by their peculiar trace element enrichment and petrographic composition; they mostly contain samples from the sites of Lipari and Stromboli, respectively. Units AIV and AVIII, restricted to the sites of Filicudi and Stromboli, show distinct petrographic characters but overlapped geochemical fingerprints.

Description: The risk of antibody-dependent enhancement (ADE) of dengue virus (DENV) infection is a major obstacle for the development of dengue vaccine candidates. Here, we described a novel approach for assessment of ADE by measuring DENV nonstructural protein 1 (NS1) production in culture supernatants with Fcγ receptor-expressing K562 cells in ELISA format (ELISA-ADE). Enhancing activities quantified by measurement of kinetics of NS1 production were in a good agreement with the results of the virus titration assay. In conjunction with the previously established enzyme-linked immunospot-based micro-neutralization test (ELISPOT-MNT) in 96-well format, the observable dose–response profiles of enhancing and neutralizing activities against all four DENV serotypes were produced with two flaviviral envelope cross-reactive monoclonal antibodies and four primary DENV-1-infected human sera. The simple high-throughput ELISA-ADE assay offers advantages for quantitative measurement of infection enhancement that can potentially be applied to large-scale seroepidemiological studies of DENV infection and vaccination.

Description: Increasing concerns over limited petroleum resources and associated environmental problems are motivating the development of efficient cell factories to produce chemicals, fuels, and materials from renewable resources in an environmentally sustainable economical manner. Bacillus spp., the best characterized Gram-positive bacteria, possesses unique advantages as a host for producing microbial enzymes and industrially important biochemicals. With appropriate modifications to heterologous protein expression and metabolic engineering, Bacillus species are favorable industrial candidates for efficiently converting renewable resources to microbial enzymes, fine chemicals, bulk chemicals, and fuels. Here, we summarize the recent advances in developing Bacillus spp. as a cell factory. We review the available genetic tools, engineering strategies, genome sequence, genome-scale structure models, proteome, and secretion pathways, and we list successful examples of enzymes and industrially important biochemicals produced by Bacillus spp. Furthermore, we highlight the limitations and challenges in developing Bacillus spp. as a robust and efficient production host, and we discuss in the context of systems and synthetic biology the emerging opportunities and future research prospects in developing Bacillus spp. as a microbial cell factory.

Description: Glucosamine (GlcN), an amino sugar, is a compound derived from substitution of a hydroxyl group of a glucose molecule with an amino group. GlcN and its acetylated derivative, N -acetylglucosamine (GlcNAc), have been widely used in food, cosmetics, and pharmaceutical industries and are currently produced by acid hydrolysis of chitin (a linear polymer of GlcNAc) extracted from crab and shrimp shells. Microbial fermentation by filamentous fungi or recombinant Escherichia coli , as an alternative method for the production of GlcN and GlcNAc, is attracting increasing attention because it is an environmentally friendly process. Although the microbial production of GlcN and GlcNAc is hampered by low yield and high production cost, considerable advances have been made in recent years. Here we review the applications, commercial market, and production of GlcN and GlcNAc, with emphasis on the metabolic and process engineering strategies used to improve GlcN and GlcNAc production by recombinant microbes.

Description: Dissemination of Della Robbia glazed terracotta in the Marche (Italy) region started from the third decade of the 16th century. Numerous altarpieces, some of which no longer exist, document this artistic production. The protagonists of this diffusion phase were two of Andrea Della Robbia’s sons, Marco (Fra Mattia) and Francesco (Fra Ambrogio). This paper shows the results of the scientific investigations carried out on constitutive materials of different altarpieces located in South Marche belonging to the Fra Mattia’s production: the Coronation of Virgin between Saints Rocco, Sebastian, Peter martyr and Antonio abbot , dated back to 1527–1530, located in the collegiate church of S. Maria Assunta in Montecassiano; the Annunciation , dated back to 1520, placed in the church of S. Maria del Soccorso in Arcevia; the fragmentary Crowned Madonna and saints altarpiece, probably realized after 1531, today preserved in Civic Museum of Ripatransone. The first altarpiece was made in Montecassiano using two different assembling or production techniques: the external part of the lunette and the pillar strips are made of glazed polychrome terracotta, while the altar step and the internal part are an interesting and uncommon example of polychrome painted terracotta. The provenance of the glazed Arcevia altarpiece is not clear yet: some historians hypothesize a local manufacture of Fra Mattia and some others a Roman or Florentine production. The remaining parts of Ripatransone altarpiece are partially glazed and partially not coated perhaps because they were unfinished and not yet painted. Clay body samples collected from the above mentioned altarpieces were investigated using different analytical techniques (OM, XRD, XRF, PIXE) to point out differences in chemical and mineralogical composition and to determine if the altarpieces were made by using local raw clay materials or other clays from Tuscany or Campania as in the Della Robbia previous production. A comparison has also been made with literature data on the Della Robbia terracotta masterpieces. From the chemical point of view, the clay bodies show a good compositional homogeneity and result calcium and iron-rich according with the raw local materials. The presence of gehlenite, pyroxenes, and hematite and the incomplete decomposition of clay minerals indicate that firing for all different terracotta parts occurred in a thermal range between 800 and 950 °C, mostly in oxidizing conditions.

Description: In this work, the successful coupling of enzymatic oxidation and aldol addition reactions for the synthesis of a Cbz-aminopolyol from a Cbz-amino alcohol was achieved for the first time in a multienzymatic one-pot system. The two-step cascade reaction consisted of the oxidation of Cbz-ethanolamine to Cbz-glycinal catalyzed by chloroperoxidase from the fungus Caldariomyces fumago and aldol addition of dihydroxyacetone phosphate to Cbz-glycinal catalyzed by rhamnulose-1-phosphate aldolase expressed as a recombinant enzyme in Escherichia coli , yielding (3 R ,4 S )-5-{[(benzyloxy)carbonyl]amino}-5-deoxy-1- O -phosphonopent-2-ulose. Tools of enzymatic immobilization, reactor configurations, and modification of the reaction medium were applied to highly increase the production of the target compound. While the use of soluble enzymes yielded only 23.6 % of Cbz-aminopolyol due to rapid enzyme inactivation, the use of immobilized ones permitted an almost complete consumption of Cbz-ethanolamine, reaching Cbz-aminopolyol yields of 69.1 and 71.9 % in the stirred-tank and packed-bed reactor, respectively. Furthermore, the reaction production was 18-fold improved when it was catalyzed by immobilized enzymes in the presence of 5 % ( v / v ) dioxane, reaching a value of 86.6 mM of Cbz-aminopoliol (31 g/L).

Description: The complete genome of Acinetobacter oleivorans DR1 contains AqsR and AqsI genes, which are LuxR and LuxI homolog, respectively. In a previous study, we demonstrated that quorum sensing (QS) signals play an important role in biofilm formation and hexadecane biodegradation. However, the regulation of genes controlled by the QS system in DR1 remains unexplored. We constructed an aqsR mutant and performed RNA sequencing analysis to understand the QS system. A total of 353 genes were differentially expressed during the stationary phase of wild-type cells compared to that of the aqsR mutant. AqsR appears to be an exceptionally important regulator because knockout of aqsR affected global gene expression. Genes involved in posttranslational modification, chaperones, cell wall structure, secondary metabolites biosynthesis, and stress defense were highly upregulated only in the wild type. Among upregulated genes, both the AOLE_03905 (putative surface adhesion protein) and the AOLE_11355 (L-asparaginase) genes have putative LuxR binding sites at their promoter regions. Soluble AqsR proteins were successfully purified in Escherichia coli harboring both aqsR and aqsI . Comparison of QS signals in an AqsI–AqsR co-overexpression strain with N -acyl homoserine lactone standards showed that the cognate N -acyl homoserine lactone binding to AqsR might be 3OH C12HSL. Our electrophoretic mobility shift assays with purified AqsR revealed direct binding of AqsR to those promoter regions. Our data showed that AqsR functions as an important regulator and is associated with several phenotypes, such as hexadecane utilization, biofilm formation, and sensitivity to cumene hydroperoxide.

Description: A membrane-bound, pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenase (ADH) was purified from Frateuria aurantia LMG 1558 T . Although F . aurantia belongs to a group of γ-Proteobacteria, the characteristics of its PQQ-ADH were similar to the enzyme characteristics of the typical high-acetic acid-resistant bacterium Gluconacetobacter europaeus from the group of α-Proteobacteria. The PQQ-dependent ADH was solubilized from the membranes and purified after anionic, cationic, and affinity chromatography with specific activity of 117 U/mg. The purified enzyme was estimated to be composed of two subunits of ca. 72 and 45 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The purified enzyme had maximum activity at pH 4.5 and showed the highest substrate specificity to ethanol, isoamyl alcohol, 1-butanol, and 1-propanol. The deduced sequences of cloned genes adhA and adhB encoding subunits I and II of PQQ-ADH showed 80 % amino acid (AA) identity to AdhA and 68 % AA identity to AdhB of Ga . europaeus V3 (LMG 18494). Because of the high similarity between genes encoding subunits I and II of PQQ-ADH and its homologous genes found in a distantly related taxonomic group of acetic acid bacteria, the results suggest the possibility of horizontal gene transfer between these two groups of genera.

Description: Enantioselective oxidation of racemic phenyl-1,2-ethanediol into ( R )-(−)-mandelic acid by a newly isolated Brevibacterium lutescens CCZU12-1 was demonstrated. It was found that optically active ( R )-(−)-mandelic acid ( e.e.p  〉 99.9 %) is produced leaving the other enantiomer ( S )-(+)-phenyl-1,2-ethanediol intact. Using fed-batch method, a total of 172.9 mM ( R )-(−)-mandelic acid accumulated in the reaction mixture after the seventh feed. Moreover, oxidation of phenyl-1,2-ethanediol using calcium alginate-entrapped resting cells was carried out in the aqueous system, and efficient biocatalyst recycling was achieved as a result of cell immobilization in calcium alginate, with a product-to-biocatalyst ratio of 27.94 g ( R )-(−)-mandelic acid g −1 dry cell weight cell after 16 cycles of repeated use.

Description: The bacterium Bacillus amyloliquefaciens anti-CA isolated from mangrove system was found to be able to actively kill Candida albicans isolated from clinic. The bacterial strain anti-CA could produce high level of bioactive substance, amylase and protease in the cheap medium containing 2.0 % soybean meal, 2.0 % wheat flour, pH 6.5 within 26 h. After purification, the main bioactive substance was confirmed to be a cyclic lipopeptide containing a heptapeptide, L-Asp→L-Leu→L-Leu→L-Val→L-Val→L-Glu→L-Leu and a 3-OH fatty acid (15 carbons). In addition to C. albicans , the purified lipopeptide can also kill many yeast strains including Metschnikowia bicuspidata , Candida tropicalis , Yarrowia lipolytica and Saccharomyces cerevisiae. After treated by the purified lipopeptide, both the whole cells and protoplasts of C. albicans were destroyed.

Description: Optical and biomedical properties of diamond-like carbon (DLC) films of various sp 2 , sp 3 bonds were studied. The layers were prepared by pulsed laser deposition (PLD) for laser energy densities from 4 J cm −2 to 14 J cm −2 . The percentage of sp 2 and sp 3 bonds was calculated using X-ray photoelectron spectroscopy (XPS). In dependence on density the films contained up to 70 % of sp 3 bonds. Optical properties were measured using spectroscopic ellipsometry in region from 250 nm to 1000 nm ( n =2.6–2.7; k =0.07–0.25) and by transmission measurement (from 200 nm to 1100 nm). The adhesion and growth of human fibroblasts and keratinocytes of DLC films were tested in vitro.

Description: Bubbles generated in water by focusing femtosecond and picosecond laser pulses in the presence of 100 nm gold nanoparticles have been investigated in the fluence range usually used for efficient cell transfection (100–200 mJ/cm 2 ). Since resulting bubbles are at the nanoscale, direct observation using optical microscopy is not possible. An optical in-situ method has been developed to monitor the time-resolved variation in the extinction cross-section of an irradiated nanoparticle solution sample. This method is used to measure the bubbles lifetime and deduce their average diameter. We show that bubbles generated with femtosecond pulses (40–500 fs) last two times longer and are larger in average than those generated with picosecond pulses (0.5–5 ps). Controlling those bubble properties is necessary for optimizing off-resonance plasmonic enhanced ultrafast laser cell transfection.

Description: To determine experimental conditions suitable for isotope analysis, we studied the plume dynamics of uranium. A uranium oxide sample was ablated by 2nd harmonic radiation from a Nd:YAG laser at a fluence of 0.5 J/cm 2 . The temporal evolution of the ablation plume was investigated in vacuum and helium environments. In vacuum, the flow velocity perpendicular to the sample surface was determined to be 2.7 km/s for neutral atoms and 4.0 km/s for singly charged atoms. These velocities are about 20 % lower than those of cerium measured under similar conditions. From the evolution of the plume in helium, we found that an observation time of 3–5 μs and an observation height of about 2.5 mm are most suited for obtaining higher sensitivity. Observation times less than 3 μs were unsuitable for precise isotope analysis since the spectral modifications arising from the Doppler splitting effect are different between the two uranium isotopes. Using the established conditions, we evaluated the calibration curve linearity, limit of detection, and precision for three samples having different abundances of  235 U.

Description: Nondiffracting Bessel beams have been recently applied to single shot ultrafast laser processing of high aspect ratio nanochannels. We review the principles and benefits of nondiffracting Bessel beams for material processing, and we compare experimental results with numerical simulations. We show how the stationarity of nonlinear propagation of ultrafast laser pulses allows high aspect ratio material processing.

Description: Electrical conductivity, Hall effect and magnetoresistance of In 2 O 3 :Sn thin films deposited on a glass substrates at different temperatures and oxygen pressures, have been investigated in the temperature range 4.2–300 K. The observed temperature dependences of resistivity for films deposited at 230 °C as well as at nominally room temperatures were typical for metallic transport of electrons except temperature dependence of resistivity of the In 2 O 3 :Sn film deposited in the oxygen deficient atmosphere. The electrical measurements were accompanied by AFM and SEM studies of structural properties, as well as by XPS analysis. It is established that changes of morphology and crystallinity of ITO films modify the low-temperature behavior of resistivity, which still remains typical for metallic transport. This is not the case for the oxygen deficient ITO layer. XPS analysis shows that grown in situ oxygen deficient ITO films have enhanced DOS between the Fermi level and the valence band edge. The extra localized states behave as acceptors leading to a compensation of n -type ITO. That can explain lower n -type conductivity in this material crossing over to a Mott-type hopping at low temperatures. Results for the low temperature measurements of stoichiometric ITO layers indicate that they do not show any trace of metal-to-insulator transition even at 4.2 K. We conclude that, although ITO is considered as a highly doped wide-band gap semiconductor, its low-temperature properties are very different from those of conventional highly doped semiconductors.

Description: The epothilones, compounds with anticancer mechanisms similar to that of paclitaxel (Taxol), are produced by strains of the myxobacterium Sorangium cellulosum , and the gene cluster responsible for epothilone biosynthesis is organised as a large operon. In this work, we showed that the 440-bp promoter regions of the operons from eight S. cellulosum strains have 94.27 % DNA sequence identity and 50 % variability in promoter activity in Escherichia coli . A primer extension analysis revealed two transcriptional start sites (TSSs) at 246 (TSS1) and 193 bp (TSS2) upstream of the translation start site (TLS), respectively. Promoter truncation determined that the basal promoter from the So0157-2 strain is located within a 264-bp region containing weak promoter activity; whereas in the 38-bp region upstream, the 264-bp promoter was required for the strong promoter activity, which was dramatically increased by 11-fold in average. There was a conserved stem–loop structure between TSS2 and the TLS, which was identified in E. coli as a negative regulatory element. In addition, the upstream non-conserved 357-bp non-coding region contributes to the promoter activity, increasing it by 1.5-fold. In conclusion, the expression of the epothilone operon non-coding region in E. coli is regulated by a double promoter (with −35 and −10 regions and two distinct TSSs), a stem–loop structure, and a distal non-coding region.

Description: Bacterial community compositions were characterized using denaturing gradient gel electrophoresis analysis of bacterial 16S rRNA gene in the sediments of the Pearl River estuary. Sequencing analyses of the excised bands indicated that Gram-negative bacteria, especially Gammaproteobacteria , were dominant in the Pearl River estuary. The diversity of polycyclic aromatic hydrocarbon ring-hydroxylating dioxygenase (PAH-RHD) gene in this estuary was then assessed by clone library analysis. The phylogenetic analyses showed that all PAH-RHD gene sequences of Gram-negative bacteria (PAH-RHD [GN] ) were closely related to the nagAc gene described for Ralstonia sp. U2 or nahAc gene for Pseudomonas sp. 9816–4, while the PAH-RHD gene sequences of Gram-positive bacteria (PAH-RHD [GP] ) at sampling site A1 showed high sequence similarity to the nidA gene from Mycobacterium species. Meanwhile, molecular diversity of the two functional genes was higher at the upstream of this region, while lower at the downstream. Redundancy analysis indicated that environmental factors, such as NH 4 -N, ∑PAHs, pH, SiO 3 -Si, and water depth, affected the distribution of the PAH-RHD [GN] gene in the Pearl River estuary.

Description: In order to provide a more suitable response to public health concerns, we improved the detection of infectious human adenoviruses in water by optimising the commonly used integrated cell culture–PCR method. Risk evaluation studies seek for rapid detection of infectious adenoviruses, including the enteric types 40 and 41 that are considered as the second most common agents of gastroenteritis in children next to rotaviruses. The here-employed 293A cell line used for infectious status assessment showed its ability to multiply adenoviruses including type 41. Two modifications were moreover applied to the workflow for viral detection. The first occurred at the nucleic acid extraction step performed directly on all infected cells, while the second was the application of real-time quantitative PCR as detection tool. All adaptations led to a 3-day reduction of the response delay and an improved sensitivity especially for the enteric adenoviral types. The infectious status of laboratory strain types 2 and 41 was demonstrated by a more than 2-log 10 increase in genome quantity. These conclusions were confirmed and reinforced by the analysis of water samples applying the improved assay. Naturally occurring infectious adenoviruses were detected in wastewater and river water, within 2 days. Types belonging to the species human adenoviruses C and type 31 were observed, but the most frequently identified type was 41 (71 % of identified sequences, n  = 34). This highlights the usefulness of our method for a wide range of types, and especially for the most prevalent and public health-relevant enteric adenoviruses.

Description: Proteins with a Zn(II) 2 Cys 6 domain, Cys-X 2 -Cys-X 6 -Cys-X 5-12 -Cys-X 2 -Cys-X 6-9 -Cys (hereafter, referred to as the C6 domain), form a subclass of zinc finger proteins found exclusively in fungi and yeast. Genome sequence databases of Saccharomyces cerevisiae and Candida albicans have provided an overview of this family of genes. Annotation of this gene family in most fungal genomes is still far from perfect and refined bioinformatic algorithms are urgently needed. Aspergillus flavus is a saprophytic soil fungus that can produce the carcinogenic aflatoxin. It is the second leading causative agent of invasive aspergillosis. The 37-Mb genome of A . flavus is predicted to encode 12,000 proteins. Two and a half percent of the total proteins are estimated to contain the C6 domain, more than twofold greater than those estimated for yeast, which is about 1 %. The variability in the spacing between cysteines, C 3 -C 4 and C 5 -C 6 , in the zinc cluster enables classification of the domains into distinct subgroups, which are also well conserved in Aspergillus nidulans . Sixty-six percent (202/306) of the A . flavus C6 proteins contain a specific transcription factor domain, and 7 % contain a domain of unknown function, DUF3468. Two A . nidulans C6 proteins containing the DUF3468 are involved in asexual conidiation and another two in sexual differentiation. In the anamorphic A . flavus , a homolog of the latter lacks the C6 domain. A . flavus being heterothallic and reproducing mainly through conidiation appears to have lost some components involved in homothallic sexual development. Of the 55 predicted gene clusters thought to be involved in production of secondary metabolites, only about half have a C6-encoding gene in or near the gene clusters. The features revealed by the A . flavus C6 proteins likely are common for other ascomycete fungi.

Description: A range of isolates of Pseudomonas aeruginosa from widely different environmental sources were examined for their ability to synthesise rhamnolipid biosurfactants. No significant differences in the quantity or composition of the rhamnolipid congeners could be produced by manipulating the growth conditions. Sequences for the rhamnolipid genes indicated low levels of strain variation, and the majority of polymorphisms did lead to amino acid sequence changes that had no evident phenotypic effect. Expression of the rhlB and rhlC rhamnosyltransferase genes showed a fixed sequential expression pattern during growth, and no significant up-regulation could be induced by varying producer strains or growth media. The results indicated that rhamnolipids are highly conserved molecules and that their gene expression has a rather stringent control. This leaves little opportunity to manipulate and greatly increase the yield of rhamnolipids from strains of P. aeruginosa for biotechnological applications.

Description: Streptococcus alactolyticus strain FGM, isolated from chicken cecum, was used to increase the extract yield of polysaccharides during Astragalus membranaceus fermentation. It was previously demonstrated that polysaccharides from fermented A . membranaceus by S . alactolyticus had some similar properties to those from A . membranaceus in terms of its ability to help heal hepatic fibrosis in rats and modulate immunopotentiation of broiler chicken. However, methods to increase the yield of the polysaccharides during fermentation of A . membranaceus are not well understood. In this paper, we investigated the involvement of uridine diphosphate (UDP)-glucose 4-epimerase ( galE ) and glucan-1,6-α-glucosidase ( dexB ) during A . membranaceus fermentation through real-time reverse transcription quantitative PCR. The galE and dexB genes of S . alactolyticus were cloned by homology-based cloning and the genome walking method for the first time, and the 3D structure of dexB was analyzed by Swiss-PdbViewer 4.0.1 software. The expression of both the galE and dexB genes in A . membranaceus fermentation was studied using the determined ideal reference gene ldh for transcript normalization. The results showed that these two genes were both highly induced and peaked after 12 h of fermentation. The expression level of galE was stepwise increased from 48 to 72 h, while dexB transcripts were markedly increased at 60 h and decreased by 72 h. These data suggested that dexB and galE of S . alactolyticus strain FGM were involved in the regulation of A . membranaceus fermentation and they might play some roles in the increase of polysaccharides.

Description: Ion implantation was used to locally modify the surface of silica glass to create periodic plasmonic microstructures with Cu nanoparticles. Nanoparticles were synthesized by Cu-ion irradiation of the silica glass at the ion energy of 40 keV, dose of 5×10 16  ions/cm 2 and current density of 5 μA/cm 2 . This procedure involves low-energy ion implantation into the glass through a mask placed at the surface. Formation of nanoparticles was observed by optical spectroscopy and atomic force microscopy. The presented results clearly demonstrate how the low-energy ions can be used for the fabrication of photonic microstructures on dielectric surfaces in a single-step process.

Description: Au/Nb:SrTiO 3 /Ti structures were fabricated by depositing Au and Ti electrodes on a single crystal 0.5 wt% Nb:SrTiO 3 (NSTO) using rf-magnetron sputtering technique. Resistive switching properties at different temperature were investigated. The Ti/NSTO interface was ohmic contact, which indicated that the resistive switching behavior was attributed to Au/NSTO interface. The resistive switching behavior happened only at the temperature above 180 K, which was possibly caused by the increase of Schottky barrier height with the increase of temperature. The structure showed a semiconductor behavior at high-resistance state (HRS) and a metallic behavior at low-resistance state (LRS). The switching conduction mechanism of Au/NSTO/Ti device is primarily described as space-charge-limited conduction (SCLC) according to the electrical transport properties measurement.

Description: The ZnO nanowire (NW) array/TiO 2 nanoparticle (NP) composite photoelectrode with controllable NW aspect ratio has been grown from aqueous solutions for the fabrication of dye-sensitized solar cells (DSSCs), which combines the advantages of the rapid electron transport in ZnO NW array and the high surface area of TiO 2 NPs. The results indicate that the composite photoelectrode achieves higher overall photoelectrical conversion efficiency ( η ) than the ZnO NW alone. As a result, DSSCs based on the ZnO NW array/TiO 2 NP composite photoelectrodes get the enhanced photoelectrical conversion efficiency, and the highest η is also achieved by rational tuning the aspect ratio of ZnO NWs. With the proper aspect ratio (ca. 6) of ZnO NW, the ZnO NW array/TiO 2 NP composite DSSC exhibits the highest conversion efficiency (5.5 %). It is elucidated by the dye adsorption amount and interfacial electron transport of DSSCs with the ZnO NW array/TiO 2 NP composite photoelectrode, which is quantitatively characterized using the UV-Vis absorption spectra and electrochemical impedance spectra. It is evident that the DSSC with the proper aspect ratio of ZnO NW displays the high dye adsorption amount and fastest interfacial electron transfer.

Description: We report the photoluminescence eigenmodes of the ZnO semiconductor microcavity on a Ag/Si substrate at room temperature. The experiment results show that the photoluminescence modes do not depend on the excitation intensity. The eigenmode in the microcavity is considered to be the perpendicular Fabry–Perot modes, where the effect of the strong coupling between plasmon and exciton is observed with the reducing of the effective refractive index from original 2.0 to 1.67 at the wavelength of 585 nm. Meanwhile, there is also a coupling between the plasmon and the exciton-polariton in the band-edge region, and the effective refractive index 1.92 is in good agreement between the experimental and the theoretical results.

Description: Variations in the switching threshold voltage of memristive devices present significant challenges for their integration into large-scale circuits. In this paper, we propose to address this problem by adding a device exhibiting S-type (N-type) negative differential resistance (NDR) in series (parallel) with memristive devices. The main effect comes from the transition between low- and high-conductivity branches of the NDR device, which leads to a redistribution of the voltage drop inside the device stack, and, as a result, the effective lowering of variations in the switching threshold. The idea is checked experimentally using a TiO 2− x memristive device connected in parallel with a tunnel GaAs diode.

Description: The detailed chemical analysis of fossils has the potential to reveal great insight to the composition, preservation and biochemistry of ancient life. Such analyses would ideally identify, quantify, and spatially resolve the chemical composition of preserved bone and soft tissue structures, but also the embedding matrix. Mapping the chemistry of a fossil in situ can place constraints on mass transfer between the enclosing matrix and the preserved organism(s), and therefore aid in distinguishing taphonomic processes from original chemical zonation remnant within the fossils themselves. Conventional analytical methods, such as scanning electron microscopy (SEM) and pyrolysis–gas chromatography/mass spectrometry (Py-GC/MS) have serious limitations in this case, primarily, an inability to provide large (i.e., decimeter) scale chemical maps. Additionally, vacuum chamber size and the need for destructive sampling preclude analysis of large and precious fossil specimens. However, the recent development of Synchrotron Rapid Scanning X-ray Fluorescence (SRS-XRF) at the Stanford Synchrotron Radiation Lightsource (SSRL) allows the non-destructive chemical analysis and imaging of major, minor, and trace element concentrations of large paleontological and archeological specimens in rapid scanning times. Here we present elemental maps of a fossil reptile produced using the new SRS-XRF method. Our results unequivocally show that preserved biological structures are not simply impressions or carbonized remains, but possess a remnant of the original organismal biochemistry. We show that SRS-XRF is a powerful new tool for the study of paleontological and archaeological samples.

Description: Here for the first time we describe the use of high resolution nanoprobe X-ray fluorescence (XRF) mapping for the analysis of artists’ paints, hierarchically complex materials typically composed of binder, pigments, fillers, and other additives. The work undertaken at the nanoprobe sought to obtain highly spatially resolved, highly sensitive mapping of metal impurities (Pb, Cd, Fe, and other metals) in submicron particles of zinc oxide pigments used in early 20th century artists’ tube paints and enamel paints, with particular emphasis on Ripolin, a popular brand of French house paint used extensively by Pablo Picasso and some of his contemporaries. Analysis revealed that the Zn oxide particles only contain a little Fe, proving that the highest quality Zn oxide pigment, free of Pb and Cd, was used for Ripolin house paints as well as artists’ paints. Nanoprobe XRF mapping also demonstrated that artists’ tube paints generally have more abundant fillers and additional whites (based on Pb, Ti, Ca) than Ripolin paints, which contain mostly pure zinc oxide. The chemical characterization of paints at the nanoscale opens the path to a better understanding of their fabrication and chemical reactivity.

Description: We fabricated sulfur-doped black silicon by metal-assist chemical etching (MCE) and ion implanting. The morphologies of silicon nanowire (SiNW) arrays and the concentration of sulfur in black silicon were analyzed by scanning electron microscope (SEM). Sulfur-doped black silicon shows higher absorption in entire 0.3–2.5 μm wavelength range as compared to undoped SiNW arrays and flat silicon. The changes in the absorption spectra of black silicon with different etching durations and annealing temperature are also shown. Upon annealing, the absorption decreases significantly in 2–2.5 μm wavelength region. The novel results clearly indicate that sulfur implanting could produce below band gap absorption in the silicon substrate.

Description: Rhodococcus erythropolis WZ010 was capable of producing optically pure (2 S ,3 S )-2,3-butanediol in alcoholic fermentation. The gene encoding an acetoin(diacetyl) reductase from R. erythropolis WZ010 (ReADR) was cloned, overexpressed in Escherichia coli , and subsequently purified by Ni-affinity chromatography. ReADR in the native form appeared to be a homodimer with a calculated subunit size of 26,864, belonging to the family of the short-chain dehydrogenase/reductases. The enzyme accepted a broad range of substrates including aliphatic and aryl alcohols, aldehydes, and ketones. It exhibited remarkable tolerance to dimethyl sulfoxide (DMSO) and retained 53.6 % of the initial activity after 4 h incubation with 30 % ( v / v ) DMSO. The enzyme displayed absolute stereospecificity in the reduction of diacetyl to (2 S ,3 S )-2,3-butanediol via ( S )-acetoin. The optimal pH and temperature for diacetyl reduction were pH 7.0 and 30 °C, whereas those for (2 S ,3 S )-2,3-butanediol oxidation were pH 9.5 and 25 °C. Under the optimized conditions, the activity of diacetyl reduction was 11.9-fold higher than that of (2 S ,3 S )-2,3-butanediol oxidation. Kinetic parameters of the enzyme showed lower K m values and higher catalytic efficiency for diacetyl and NADH in comparison to those for (2 S ,3 S )-2,3-butanediol and NAD + , suggesting its physiological role in favor of (2 S ,3 S )-2,3-butanediol formation. Interestingly, the enzyme showed higher catalytic efficiency for ( S )-1-phenylethanol oxidation than that for acetophenone reduction. ReADR-catalyzed asymmetric reduction of diacetyl was coupled with stereoselective oxidation of 1-phenylethanol, which simultaneously formed both (2 S ,3 S )-2,3-butanediol and ( R )-1-phenylethanol in great conversions and enantiomeric excess values.

Description: The wettability of solid surface is a very important property of materials, which depends on both surface chemical composition and the geometry of the solid surface. We herein report a simple and inexpensive method to fabricate superhydrophobic surfaces with the use of mullite whisks (MWs) for the first time. Binary micro-/nano-structural surfaces were created by the incorporation of hydrophilic needlelike MWs with poly( γ -methacryloxypropyltrimethoxysilane) to exhibit hydrophobic behavior, which further showed superhydrophobicity with water contact angle of 152.4±2 ∘ after modified with stearic acid. The wettability variation from hydrophilicity to hydrophobicity and superhydrophobicity during the assembly process are discussed based on the scanning electron microscopy, X-ray diffraction, thermogravimetric analysis, and contact-angle system. The superhydrophobicity of the surface could be attributed to the hierarchical microstructure of the rough surface induced by MWs and low surface energy of stearic acid.

Description: Luminescent properties of ZnO nanorods covered with Ag nanoparticles are examined. Nanorods were synthesized on AAO templates using Atomic Layer Deposition (ALD) technique. Two types of the samples were prepared with different arrangement of ZnO nanorods and doping conditions. Nanorods of the second type were codoped with Al, to stimulate defect-related emissions. The ZnO material fills heterogeneously the interior of the AAO nanopores and has hexagonal, wurtzite structure. Both types of structures exhibit a broad defect-related emission at about 440 nm, most probably related to recombination at zinc interstitial (Zn i ) defects. This emission in samples with a random distribution of ZnO:Al nanorods and finer Ag nanoparticles is enhanced by factor of ∼2.5 upon Ag deposition. The so-obtained material is interesting from the point of view of its application in blue range emitting diodes.

Description: The separation mechanism of opaque and transparent model micro-particles, graphite and polystyrene copolymer spheres, respectively, from polymethyl methacrylate (PMMA) substrates were investigated employing a ns-pulse laser radiating at 532 nm. The particles transparent in the visible wavelength range could be removed from PMMA efficiently in a very narrow fluence range between 1 and 2 J/cm 2 according to a simple 1D thermal expansion model. Above this fluence region, with single pulses, the transparent microspheres caused local ablation of the PMMA substrate in the optical microlens nearfield. This process led to removal of the particles themselves due to the expansion of the ablation plasma. The irregularly shaped graphite particles shaded the underlying substrate from the incoming radiation so that no optical nearfield damage mechanism could be observed. Therefore, a substantial cleaning window between 0.5 and more than 16 J/cm 2 was provided. The graphite data suggest an ablation mechanism of the particulates themselves due to a high optical absorption coefficient.

Description: Regioselective glycosylation of flavonoids cannot be easily achieved due to the presence of several hydroxyl groups in flavonoids. This hurdle could be overcome by employing uridine diphosphate-dependent glycosyltransferases (UGTs), which use nucleotide sugars as sugar donors and diverse compounds including flavonoids as sugar acceptors. Quercetin rhamnosides contain antiviral activity. Two quercetin diglycosides, quercetin 3- O -glucoside-7- O -rhamnoside and quercetin 3,7- O -bisrhamnoside, were synthesized using Escherichia coli expressing two UGTs. For the synthesis of quercetin 3- O -glucoside-7- O -rhamnoside, AtUGT78D2, which transfers glucose from UDP-glucose to the 3-hydroxyl group of quercetin, and AtUGT89C1, which transfers rhamnose from UDP-rhamnose to the 7-hydroxyl group of quercetin 3- O -glucoside, were transformed into E. coli . Using this approach, 67 mg/L of quercetin 3- O -glucoside-7- O -rhamnoside was synthesized. For the synthesis of quercetin 3,7- O -bisrhamnoside, AtUGT78D1, which transfers rhamnose to the 3-hydroxy group of quercetin, and AtUGT89C1 were used. The RHM2 gene from Arabidopsis thaliana was coexpressed to supply the sugar donor, UDP-rhamnose. E. coli expressing AtUGT78D1 , AtUGT89C1 , and RHM2 was used to obtain 67.4 mg/L of quercetin 3,7- O -bisrhamnoside.

Description: Biological soil disinfestations (BSDs) were developed separately in Japan and in The Netherlands as an alternative to chemical fumigations. In Japan, it was developed based on the knowledge of irrigated paddy rice and upland crop rotation system that was rather tolerant of soil-borne disease development. The methods consist of application of easily decomposable organic matter, irrigation, and covering the soil surface with plastic film, thereby inducing anaerobic (reductive) soil conditions and suppressing many soil-borne pests including fungi, bacteria, nematodes, and weeds. The methods are widely used by organic farmers in the area where residences and agricultural fields are intermingled. To note one advantage of these methods, maintenance of soil suppressiveness to Fusarium wilt of tomato was suggested, while soil treated with chloropicrin became conducive to the disease. Suppression of soil-borne fungal pathogens by BSDs might be attributed to anaerobicity and high temperature, organic acids generated, and metal ions released into soil water. Contributions of respective factors to suppression of respective pathogens might be diverse. Presumably, these factors might vary on the fungal community structure in BSD-treated soil. These factors also work in paddy fields. Therefore, the BSDs developed in Japan are probably a method to raise the efficacy of paddy–upland rotation through intensive organic matter application and through maintenance of a strongly anaerobic (reductive) soil condition.

Description: The human host cell line, F2N78, is a new somatic hybrid cell line designed for therapeutic antibody production. To verify its potential as a human host cell line, recombinant F2N78 cells that produce antibody against rabies virus (rF2N78) were cultivated at different culture pH (6.8, 7.0, 7.2, 7.4, and 7.6) and temperatures (33.0 °C and 37.0 °C). Regardless of the culture temperature, the highest specific growth rate was obtained at a pH of 7.0–7.4. Lowering the culture temperature from 37.0 °C to 33.0 °C suppressed cell growth while allowing maintenance of high cell viability for a longer period. However, it did not enhance antibody production because specific antibody productivity did not increase at 33.0 °C. The highest maximum antibody concentration was obtained at 37.0 °C and pH 6.8. The N-linked glycosylation of the antibody was affected by the culture pH rather than the temperature. Nevertheless, G1F was dominant and G2F occupied a larger portion than G0F in all culture conditions. Compared to the same antibody produced from recombinant CHO cells, the antibody produced from rF2N78 cells has more galactose capping and was more similar to human plasma IgG. Taken together, the results obtained here demonstrate the potential of F2N78 as an alternative human host cell line for therapeutic antibody production.

Description: Microbial enhanced oil recovery (MEOR) refers to the process of using bacterial activities for more oil recovery from oil reservoirs mainly by interfacial tension reduction and wettability alteration mechanisms. Investigating the impact of these two mechanisms on enhanced oil recovery during MEOR process is the main objective of this work. Different analytical methods such as oil spreading and surface activity measurements were utilized to screen the biosurfactant-producing bacteria isolated from the brine of a specific oil reservoir located in the southwest of Iran. The isolates identified by 16S rDNA and biochemical analysis as Enterobacter cloacae (Persian Type Culture Collection (PTCC) 1798) and Enterobacter hormaechei (PTCC 1799) produce 1.53 g/l of biosurfactant. The produced biosurfactant caused substantial surface tension reduction of the growth medium and interfacial tension reduction between oil and brine to 31 and 3.2 mN/m from the original value of 72 and 29 mN/m, respectively. A novel set of core flooding tests, including in situ and ex situ scenarios, was designed to explore the potential of the isolated consortium as an agent for MEOR process. Besides, the individual effects of wettability alteration and IFT reduction on oil recovery efficiency by this process were investigated. The results show that the wettability alteration of the reservoir rock toward neutrally wet condition in the course of the adsorption of bacteria cells and biofilm formation are the dominant mechanisms on the improvement of oil recovery efficiency.

Description: Bioaugmentation with degrading bacteria is an effective method to improve the treatment of refractory industrial wastewater; nevertheless there were controversial opinions about the fate of inoculated bacteria and microbial community dynamics. In this study, two lab-scale sequencing batch reactors filled with modified zeolite were used to treat a coking wastewater with pyridine and quinoline shock load, and a bacterial consortium containing three degrading strains was added in one reactor for bioaugmentation. During 120-day operation, the bioaugmented reactor removed over 99 % pyridine, 99 % quinoline, 85 % TOC, 65 % COD, and 95 % NO 3 − -N with higher resistance to the shock load than the non-bioaugmented reactor. Based on the terminal restriction fragment length polymorphism of 16S rDNA, bacterial community diversity increased in the bioaugmented reactor. Principal component analysis revealed that, to cope with the shock load, the indigenous bacterial community recovered to the initial structure by acclimatizing itself constantly to the inhospitable environment; but bioaugmentation accelerated the shift of whole bacterial community, resulting in a far different structure from the initial one. Canonical correspondence analysis indicated that the environmental parameters of pyridine, quinoline, TOC, and NO 3 − -N had close negative correlations with bioaugmentation; and NH 3 -N and COD were the main parameters to impact on the bacterial community changes and treatment efficiency.

Description: Pseudomonads are metabolically versatile microbes that employ complex regulatory networks to control gene expression, particularly with respect to carbon and nitrogen metabolism. The aim of this study was to characterise the regulatory networks that control pyrimidine metabolism (hydantoin-hydrolysing activity) in Pseudomonas putida strain RU-KM3 S , focussing on transcriptional activation of dihydropyrimidinase (Dhp) and β-ureidopropionase (Bup), encoding dhp and bup , respectively. The two genes are arranged divergently on the chromosome and are separated by ORF1, encoding a putative transporter, which lies upstream of and in the same orientation as bup . The results from this study reveal that pyrimidine metabolism, as a function of Bup and Dhp activity in P . putida RU-KM3 S , is controlled by a complex regulatory network including several global pathways in addition to induction by the substrate. Three major control pathways act at the level of transcriptional and include: (1) induction of transcriptional activation in the presence of hydantoin, (2) carbon catabolite repression mediated via a pathway independent of Crc and (3) quorum sensing that does not require a putative lux box located upstream of the dhp transcriptional start. Finally, the data suggest a minor role for the global regulators Anr, Vfr and Crc, likely through regulation of the activity of transcription factors interacting directly with the bup /ORF1– dhp promoter.

Description: Bacillus subtilis ferments pyruvate to 2,3-butanediol via α-acetolactate synthase, α-acetolactate decarboxylase, and butanediol dehydrogenase (BDH), encoded by the alsSD operon and the unlinked monocistronic bdhA gene, respectively. Upstream and divergent from alsSD is the alsR gene that encodes AlsR, a member of the LysR-type transcriptional regulator family. AlsR directly stimulates alsSD transcription by binding to characteristic sites preceding the alsS promoter, but its effect on bdhA expression was unknown. The effect of AlsR on bdhA expression was assessed in a wild-type strain and a congenic strain carrying an alsR::spc knockout mutation by measuring: (a) expression of a transcriptional bdhA-lacZ fusion; (b) bdhA mRNA steady-state levels by quantitative reverse transcriptase PCR; and (c) expression of BDH enzymatic activity. Activation of bdhA expression occurred in early stationary phase, and expression was lowered, but not abolished, in the alsR::spc mutant. Mapping the transcriptional start site of bdhA by primer extension revealed a 268-nucleotide 5′-untranslated region preceding the bdhA initiation methionine codon. Transcription initiation was not reduced in the alsR::spc mutant, and by electrophoretic mobility shift assay, purified AlsR protein did not bind to the bdhA promoter region, suggesting that bdhA expression is indirectly under AlsR transcriptional control.