The Role of Liquid Ink Transport in the Direct Placement of Quantum Dot Emitters onto Sub-Micrometer Antennas by Dip-Pen Nanolithography.

Dip-pen nanolithography (DPN) is used to precisely position core/thick-shell ("giant") quantum dots (gQDs; ≥10 nm in diameter) exclusively on top of silicon nanodisk antennas (≈500 nm diameter pillars with a height of ≈200 nm), resulting in periodic arrays of hybrid nanostructures and demonstrating a facile integration strategy toward next-generation quantum light sources. A three-step reading-inking-writing approach is employed, where atomic force microscopy (AFM) images of the pre-patterned substrate topography are used as maps to direct accurate placement of nanocrystals. The DPN "ink" comprises gQDs suspended in a non-aqueous carrier solvent, o-dichlorobenzene. Systematic analyses of factors influencing deposition rate for this non-conventional DPN ink are described for flat substrates and used to establish the conditions required to achieve small (sub-500 nm) feature sizes, namely: dwell time, ink-substrate contact angle and ink volume. Finally, it is shown that the rate of solvent transport controls the feature size in which gQDs are found on the substrate, but also that the number and consistency of nanocrystals deposited depends on the stability of the gQD suspension. Overall, the results lay the groundwork for expanded use of nanocrystal liquid inks and DPN for fabrication of multi-component nanostructures that are challenging to create using traditional lithographic techniques.

Template-Based Orbital Wall Fracture Treatment Using Statistical Shape Analysis.

PURPOSE: Aim of this study was to investigate whether a mold generated from a statistical shape model of the orbit could be generated to provide a cost-efficient means for the treatment of orbital fractures. MATERIALS AND METHODS: A statistical shape model was created from 131 computed tomographic (CT) scans of unaffected adult middle European human orbits. To generate the model, CT scans were segmented in Brainlab software, preregistered using anatomic landmarks, trimmed to an identical size, and definitely registered. Then, the model was created using the global master algorithm. Based on this model, a mold consisting of a male part and a female part was constructed and printed using a rapid prototyping technique. RESULTS: A statistical shape model of the human orbit was generated from 125 CT scans. Six scans (4.5%) presented major anatomic deviations and were discarded. A solid mold based on this model was printed. Using this mold, flat titanium mesh could be successfully deformed to serve as an orbital implant. CONCLUSION: A mold based on the statistical orbital shape could serve as a cost-effective means for the treatment of orbital fractures. It allows the anatomic preformation of titanium or resorbable implant material for orbital reconstruction. Because these materials could be cut from larger sheets, the use of a mold would be a cost-effective treatment alternative.

Correction of B 0-induced geometric distortion variations in prospective motion correction for 7T MRI.

OBJECTIVE: Prospective motion correction can effectively fix the imaging volume of interest. For large motion, this can lead to relative motion of coil sensitivities, distortions associated with imaging gradients and B 0 field variations. This work accounts for the B 0 field change due to subject movement, and proposes a method for correcting tissue magnetic susceptibility-related distortion in prospective motion correction. MATERIALS AND METHODS: The B 0 field shifts at the different head orientations were characterized. A volunteer performed large motion with prospective motion correction enabled. The acquired data were divided into multiple groups according to the object positions. The correction of B 0-related distortion was applied to each group of data individually via augmented sensitivity encoding with additionally integrated gradient nonlinearity correction. RESULTS: The relative motion of the gradients, B 0 field and coil sensitivities in prospective motion correction results in residual spatial distortion, blurring, and coil artifacts. These errors can be mitigated by the proposed method. Moreover, iterative conjugate gradient optimization with regularization provided superior results with smaller RMSE in comparison to standard conjugate gradient. CONCLUSION: The combined correction of B 0-related distortion and gradient nonlinearity leads to a reduction of residual motion artifacts in prospective motion correction data.

Correction of gradient nonlinearity artifacts in prospective motion correction for 7T MRI.

PURPOSE: To demonstrate the effect of gradient nonlinearity and develop a method for correction of gradient nonlinearity artifacts in prospective motion correction (Mo-Co). METHODS: Nonlinear gradients can induce geometric distortions in magnetic resonance imaging, leading to pixel shifts with errors of up to several millimeters, thereby interfering with precise localization of anatomical structures. Prospective Mo-Co has been extended by conventional gradient warp correction applied to individual phase encoding steps/groups during the reconstruction. The gradient-related displacements are approximated using spherical harmonic functions. In addition, the combination of this method with a retrospective correction of the changes in the coil sensitivity profiles relative to the object (augmented sensitivity encoding (SENSE) reconstruction) was evaluated in simulation and experimental data. RESULTS: Prospective Mo-Co under gradient fields and coils sensitivity inconsistencies results in residual blurring, spatial distortion, and coil sensitivity mismatch artifacts. These errors can be considerably mitigated by the proposed method. High image quality with very little remaining artifacts was achieved after a few iterations. The relative image errors decreased from 25.7% to below 17.3% after 10 iterations. CONCLUSION: The combined correction of gradient nonlinearity and sensitivity map variation leads to a pronounced reduction of residual motion artifacts in prospectively motion-corrected data.

Shock Hugoniot equations of state for binary ideal (toluene/fluorobenzene) and nonideal (ethanol/water) liquid mixtures.

Laser shock Hugoniot data were obtained using ultrafast dynamic ellipsometry (UDE) for both nonideal (ethanol/water solutions with mole percent χ(ethanol) = 0%, 3.4%, 5.4%, 7.5%, 9.7%, 11%, 18%, 33%, 56%, 100%) and ideal liquid mixtures (toluene/fluorobenzene solutions with mole percent χ(toluene) = 0%, 26.0%, 49.1%, 74.9%, 100%). The shock and particle velocities obtained from the UDE data were compared to the universal liquid Hugoniot (ULH) and to literature shock (plate impact) data where available. It was found that the water UDE data fit to a ULH-form equation suggests an intercept of 1.32 km/s, lower than the literature ambient sound speed in water of 1.495 km/s (Mijakovic et al. J. Mol. Liq. 2011, 164, 66-73). Similarly, the ethanol UDE data fit to a ULH-form equation suggests an intercept of 1.45 km/s, which lies above the literature ambient sound speed in ethanol of 1.14 km/s. Both the literature plate impact and UDE Hugoniot data lie below the ULH for water. Likewise, the literature plate impact and UDE Hugoniot data lie above the ULH for ethanol. The UDE Hugoniot data for the mixtures of water and ethanol cross the predictions of the ULH near the same concentration where the sound speed reaches a maximum. In contrast, the UDE data from the ideal liquids and their mixtures are well behaved and agree with ULH predictions across the concentration range. The deviations of the nonideal ethanol/water data from the ULH suggest that complex hydrogen bonding networks in ethanol/water mixtures alter the compressibility of the mixture.

Growth-coupled anaerobic production of isobutanol from glucose in minimal medium with Escherichia coli.

BACKGROUND: The microbial production of isobutanol holds promise to become a sustainable alternative to fossil-based synthesis routes for this important chemical. Escherichia coli has been considered as one production host, however, due to redox imbalance, growth-coupled anaerobic production of isobutanol from glucose in E. coli is only possible if complex media additives or small amounts of oxygen are provided. These strategies have a negative impact on product yield, productivity, reproducibility, and production costs. RESULTS: In this study, we propose a strategy based on acetate as co-substrate for resolving the redox imbalance. We constructed the E. coli background strain SB001 (ΔldhA ΔfrdA ΔpflB) with blocked pathways from glucose to alternative fermentation products but with an enabled pathway for acetate uptake and subsequent conversion to ethanol via acetyl-CoA. This strain, if equipped with the isobutanol production plasmid pIBA4, showed robust exponential growth (µ = 0.05 h-1) under anaerobic conditions in minimal glucose medium supplemented with small amounts of acetate. In small-scale batch cultivations, the strain reached a glucose uptake rate of 4.8 mmol gDW-1 h-1, a titer of 74 mM and 89% of the theoretical maximal isobutanol/glucose yield, while secreting only small amounts of ethanol synthesized from acetate. Furthermore, we show that the strain keeps a high metabolic activity also in a pulsed fed-batch bioreactor cultivation, even if cell growth is impaired by the accumulation of isobutanol in the medium. CONCLUSIONS: This study showcases the beneficial utilization of acetate as a co-substrate and redox sink to facilitate growth-coupled production of isobutanol under anaerobic conditions. This approach holds potential for other applications with different production hosts and/or substrate-product combinations.

Nannochloropsis oceanica harvested using electrocoagulation with alternative electrodes - An innovative approach on potential biomass applications.

Electrocoagulation is a promising technology to harvest microalgal biomass. However, the commonly used aluminum electrodes release undesired salts that decrease biomass value. In this study, alternative iron, zinc, and magnesium electrodes and operational parameters pH, time and current density were studied to harvest Nannochloropsis oceanica. For recovery efficiency and concentration factor the initial pH was most important using iron electrodes, while time and current density were more relevant using zinc and magnesium electrodes. Optimal parameters resulted in biomass recovery efficiencies > 95%, biomass was concentrated 2.8-7.2 times and contained 15.7-29.1% ashes. Elemental analysis revealed metal salts in harvested biomass resulting from electrode corrosion. Finally, ash contents could be reduced by 65% using EDTA as a chelating agent. The electrocoagulation harvested microalgal biomass enriched in essential metals may be a promising bioresource for agricultural growth inducers, or functional ingredients for feed.

Flashing lights affect the photophysiology and expression of carotenoid and lipid synthesis genes in Nannochloropsis gaditana.

Nannochloropsis gaditana is a promising microalga for biotechnology. One of the strategies to stimulate its full potential in metabolite production is exposure to flashing lights. Here, we report how N. gaditana adapts to different flashing light regimes (5, 50, and 500 Hz) by changing its cellular physiology and the relative expression of genes related to critical cellular functions. We analyzed the differential mRNA abundance of genes related to photosynthesis, nitrogen assimilation and biosynthesis of chlorophyll, carotenoids, lipids, fatty acids and starch. Analysis of photosynthetic efficiency and high mRNA abundance of photoprotection genes supported the inference that excess excitation energy provided by light absorbance during photosynthesis was produced under low frequency flashing lights and was dissipated by photopigments via the xanthophyll-cycle. Increased relative expression levels of genes related to the synthesis of carotenoids and chlorophyll confirmed the accumulation of photopigments previously observed at low frequency flashing lights. Higher differential mRNA abundance of genes related to the triacylglycerol biosynthesis were observed at lower frequency flashing lights, possibly triggered by a poor nitrogen assimilation caused by low mRNA abundance of a nitrate reductase gene. This study advances a new understanding of algal physiology and metabolism leading to improved cellular performance and metabolite production.

Drying Microalgae Using an Industrial Solar Dryer: A Biomass Quality Assessment.

Microalgae are considered a promising resource of proteins, lipids, carbohydrates, and other functional biomolecules for food and feed markets. Competitive drying solutions are required to meet future demands for high-quality algal biomass while ensuring proper preservation at reduced costs. Since often used drying methods, such as freeze or spray drying, are energy and time consuming, more sustainable processes remain to be developed. This study tested an indirect and hybrid solar dryer as an alternative to conventional freeze drying of industrially produced Tetraselmis chui and Nannochloropsis oceanica wet paste. The effects of the drying method on biomass quality parameters, including biochemical profiles, functional properties, and microbial safety, were assessed. No significant differences were found between the applied drying technologies for total proteins, carbohydrates, lipids, and fatty acid profiles. On the other hand, some pigments showed significant differences, displaying up to 44.5% higher contents in freeze-dried samples. Minor differences were also registered in the mineral profiles (<10%). Analyses of microbial safety and functional properties of the solar-dried biomass appear adequate for food and feed products. In conclusion, industrial solar drying is a sustainable technology with a high potential to preserve high-quality microalgal biomass for various markets at expected lower costs.

Algae as Food in Europe: An Overview of Species Diversity and Their Application.

Algae have been consumed for millennia in several parts of the world as food, food supplements, and additives, due to their unique organoleptic properties and nutritional and health benefits. Algae are sustainable sources of proteins, minerals, and fiber, with well-balanced essential amino acids, pigments, and fatty acids, among other relevant metabolites for human nutrition. This review covers the historical consumption of algae in Europe, developments in the current European market, challenges when introducing new species to the market, bottlenecks in production technology, consumer acceptance, and legislation. The current algae species that are consumed and commercialized in Europe were investigated, according to their status under the European Union (EU) Novel Food legislation, along with the market perspectives in terms of the current research and development initiatives, while evaluating the interest and potential in the European market. The regular consumption of more than 150 algae species was identified, of which only 20% are approved under the EU Novel Food legislation, which demonstrates that the current legislation is not broad enough and requires an urgent update. Finally, the potential of the European algae market growth was indicated by the analysis of the trends in research, technological advances, and market initiatives to promote algae commercialization and consumption.

Carotenoid biosynthetic gene expression, pigment and n-3 fatty acid contents in carotenoid-rich Tetraselmis striata CTP4 strains under heat stress combined with high light.

In this study, two carotenoid-rich strains of the euryhaline microalga Tetraselmis striata CTP4 were isolated by random mutagenesis combined with selection via fluorescence activated cell sorting and growth on norflurazon. Both strains, ED5 and B11, showed an up to 1.5-fold increase in carotenoid contents as compared with the wildtype, independent of the growth conditions. More specifically, violaxanthin, ß-carotene and lutein contents reached as high as 1.63, 4.20 and 3.81 mg g-1 DW, respectively. Genes coding for phytoene synthase, phytoene desaturase, lycopene-ß-cyclase and ε-ring hydroxylase involved in carotenoid biosynthesis were found to be upregulated in ED5 and B11 cells as compared to the wildtype. Both strains showed higher contents of eicosapentaenoic acid as compared with those of the wildtype, reaching up to 4.41 and 2.88 mg g-1 DW, respectively. Overall, these results highlight the complexity of changes in carotenoid biosynthesis regulation that are required to improve pigment contents in microalgae.

Flashing light emitting diodes (LEDs) induce proteins, polyunsaturated fatty acids and pigments in three microalgae.

As the periodic emission of light pulses by light emitting diodes (LEDs) is known to stimulate growth or induce high value biocompounds in microalgae, this flashing light regime was tested on growth and biochemical composition of the microalgae Nannochloropsis gaditana, Koliella antarctica and Tetraselmis chui. At low flashing light frequencies (e.g., 5 and 50 Hz, Duty cycle = 0.05), a strain-dependent growth inhibition and an accumulation of protein, polyunsaturated fatty acids, chlorophyll or carotenoids (lutein, ß-carotene, violaxanthin and neoxanthin) was observed. In addition, a 4-day application of low-frequency flashing light to concentrated cultures increased productivities of eicosapentaenoic acid (EPA) and specific carotenoids up to three-fold compared to continuous or high frequency flashing light (500 Hz, Duty cycle = 0.05). Therefore, applying low-frequency flashing light as finishing step in industrial production can increase protein, polyunsaturated fatty acids or pigment contents in biomass, leading to high-value algal products.

Navigator accuracy requirements for prospective motion correction.

Prospective motion correction in MRI is becoming increasingly popular to prevent the image artifacts that result from subject motion. Navigator information is used to update the position of the imaging volume before every spin excitation so that lines of acquired k-space data are consistent. Errors in the navigator information, however, result in residual errors in each k-space line. This paper presents an analysis linking noise in the tracking system to the power of the resulting image artifacts. An expression is formulated for the required navigator accuracy based on the properties of the imaged object and the desired resolution. Analytical results are compared with computer simulations and experimental data.

Nanoparticles Based on Hydrophobic Polysaccharide Derivatives-Formation Principles, Characterization Techniques, and Biomedical Applications.

Polysaccharide (PS) nanoparticles (NP) are fascinating materials that combine huge application potential with the unique beneficial features of natural biopolymers. Different types of PS-NP can be distinguished depending on the basic preparation principles (top-down vs bottom-up vs coating of nanomaterials) and the material from which they are obtained (native PS vs chemically modified PS derivatives vs nanocomposites). This review provides a comprehensive overview of an approach towards PS-NP that has gained rapidly increasing interest within the last decade; the nanoself-assembling of hydrophobic PS derivatives. This facile process is easy to perform and offers a broad structural diversity in terms of the PS backbone and the additional functionalities that can be introduced. Fundamental principles of different NP preparation techniques along with useful characterization methods are presented in this work. A comprehensive summary of PS-NP prepared by different techniques and with various PS backbones and types/amounts of hydrophobic substituents is given. The intention is to demonstrate how different parameters determine the size, size distribution, and zeta-potential of the particles. Moreover, application trends in biomedical areas are highlighted in which tailored functional PS-NP are evaluated and constantly developed further.

Temperature-Dependent Lipid Accumulation in the Polar Marine Microalga Chlamydomonas malina RCC2488.

The exploration of cold-adapted microalgae offers a wide range of biotechnological applications that can be used for human, animal, and environmental benefits in colder climates. Previously, when the polar marine microalga Chlamydomonas malina RCC2488 was cultivated under both nitrogen replete and depleted conditions at 8°C, it accumulated lipids and carbohydrates (up to 32 and 49%, respectively), while protein synthesis decreased (up to 15%). We hypothesized that the cultivation temperature had a more significant impact on lipid accumulation than the nitrogen availability in C. malina. Lipid accumulation was tested at three different temperatures, 4, 8, and 15°C, under nitrogen replete and depleted conditions. At 4°C under the nitrogen replete condition C. malina had the maximal biomass productivity (701.6 mg L-1 day-1). At this condition, protein content was higher than lipids and carbohydrates. The lipid fraction was mainly composed of polyunsaturated fatty acids (PUFA) in the polar lipid portion, achieving the highest PUFA productivity (122.5 mg L-1 day-1). At this temperature, under nitrogen deficiency, the accumulation of carbohydrates and neutral lipids was stimulated. At 8 and 15°C, under both nitrogen replete and depleted conditions, the lipid and carbohydrate content were higher than at 4°C, and the nitrogen stress condition did not affect the algal biochemical composition. These results suggest that C. malina is a polar marine microalga with a favorable growth temperature at 4°C and is stressed at temperatures ≥8°C, which directs the metabolism to the synthesis of lipids and carbohydrates. Nevertheless, C. malina RCC2488 is a microalga suitable for PUFA production at low temperatures with biomass productivities comparable with mesophilic strains.

Scale-up and large-scale production of Tetraselmis sp. CTP4 (Chlorophyta) for CO2 mitigation: from an agar plate to 100-m3 industrial photobioreactors.

Industrial production of novel microalgal isolates is key to improving the current portfolio of available strains that are able to grow in large-scale production systems for different biotechnological applications, including carbon mitigation. In this context, Tetraselmis sp. CTP4 was successfully scaled up from an agar plate to 35- and 100-m3 industrial scale tubular photobioreactors (PBR). Growth was performed semi-continuously for 60 days in the autumn-winter season (17th October - 14th December). Optimisation of tubular PBR operations showed that improved productivities were obtained at a culture velocity of 0.65-1.35 m s-1 and a pH set-point for CO2 injection of 8.0. Highest volumetric (0.08 ± 0.01 g L-1 d-1) and areal (20.3 ± 3.2 g m-2 d-1) biomass productivities were attained in the 100-m3 PBR compared to those of the 35-m3 PBR (0.05 ± 0.02 g L-1 d-1 and 13.5 ± 4.3 g m-2 d-1, respectively). Lipid contents were similar in both PBRs (9-10% of ash free dry weight). CO2 sequestration was followed in the 100-m3 PBR, revealing a mean CO2 mitigation efficiency of 65% and a biomass to carbon ratio of 1.80. Tetraselmis sp. CTP4 is thus a robust candidate for industrial-scale production with promising biomass productivities and photosynthetic efficiencies up to 3.5% of total solar irradiance.

Flashing LEDs for Microalgal Production.

Flashing lights are next-generation tools to mitigate light attenuation and increase the photosynthetic efficiency of microalgal cultivation systems illuminated by light-emitting diodes (LEDs). Optimal flashing light conditions depend on the reaction kinetics and properties of the linear electron transfer chain, energy dissipation, and storage mechanisms of a phototroph. In particular, extremely short and intense light flashes potentially mitigate light attenuation in photobioreactors without impairing photosynthesis. Intelligently controlling flashing light units and selecting electronic components can maximize light emission and energy efficiency. We discuss the biological, physical, and technical properties of flashing lights for algal production. We combine recent findings about photosynthetic pathways, self-shading in photobioreactors, and developments in solid-state technology towards the biotechnological application of LEDs to microalgal production.

Urban wastewater treatment by Tetraselmis sp. CTP4 (Chlorophyta).

The ability of a recent isolate, Tetraselmis sp. CTP4, for nutrient removal from sewage effluents before and after the nitrification process under batch and continuous cultivation was studied. Biomass productivities in both wastewaters were similar under continuous conditions (0.343±0.053gL-1d-1) and nutrient uptake rates were maximal 31.4±0.4mgNL-1d-1 and 6.66±1.57mgP-PO43-L-1d-1 in WW before nitrification when cultivated in batch. Among batch treatments, cellular protein, carbohydrate and lipid levels shifted with aging cultures from 71.7±6.3 to 29.2±1.2%, 17.4±7.2 to 57.2±3.9% and 10.9±1.7 to 13.7±4.7%, respectively. In contrast, CTP4 cultivated continuously in Algal medium (control) showed lower biomass productivities (0.282gVSSL-1d-1) although improved lipid content (up to 20% lipids) in batch cultivation. Overall, Tetraselmis sp. CTP4 is promising for WW treatment as a replacement of the costly nitrification process, fixating more nutrients and providing a protein and carbohydrate-rich biomass as by-product.

Advanced process for precipitation of lignin from ethanol organosolv spent liquors.

An advanced process for lignin precipitation from organosolv spent liquors based on ethanol evaporation was developed. The process avoids lignin incrustations in the reactor, enhances filterability of the precipitated lignin particles and significantly reduces the liquor mass in downstream processes. Initially, lignin solubility and softening properties were understood, quantified and exploited to design an improved precipitation process. Lignin incrustations were avoided by targeted precipitation of solid lignin at specific conditions (e.g. 100 mbar evaporation pressure, 43°C and 10%wt. of ethanol in lignin dispersion) in fed-batch operation at lab and pilot scale. As result of evaporation the mass of spent liquor was reduced by about 50%wt., thus avoiding large process streams. By controlled droplet coalescence the mean lignin particle size increased from below 10 µm to sizes larger than 10 µm improving the significantly filterability.

Isolation of a euryhaline microalgal strain, Tetraselmis sp. CTP4, as a robust feedstock for biodiesel production.

Bioprospecting for novel microalgal strains is key to improving the feasibility of microalgae-derived biodiesel production. Tetraselmis sp. CTP4 (Chlorophyta, Chlorodendrophyceae) was isolated using fluorescence activated cell sorting (FACS) in order to screen novel lipid-rich microalgae. CTP4 is a robust, euryhaline strain able to grow in seawater growth medium as well as in non-sterile urban wastewater. Because of its large cell size (9-22 µm), CTP4 settles down after a six-hour sedimentation step. This leads to a medium removal efficiency of 80%, allowing a significant decrease of biomass dewatering costs. Using a two-stage system, a 3-fold increase in lipid content (up to 33% of DW) and a 2-fold enhancement in lipid productivity (up to 52.1 mg L-1 d-1) were observed upon exposure to nutrient depletion for 7 days. The biodiesel synthesized from the lipids of CTP4 contained high levels of oleic acid (25.67% of total fatty acids content) and minor amounts of polyunsaturated fatty acids with ≥4 double bonds (<1%). As a result, this biofuel complies with most of the European (EN14214) and American (ASTM D6751) specifications, which commonly used microalgal feedstocks are usually unable to meet. In conclusion, Tetraselmis sp. CTP4 displays promising features as feedstock with lower downstream processing costs for biomass dewatering and biodiesel refining.