Microalgae-Based Biohybrid Microrobot for Accelerated Diabetic Wound Healing.

A variety of wound healing platforms have been proposed to alleviate the hypoxic condition and/or to modulate the immune responses for the treatment of chronic wounds in diabetes. However, these platforms with the passive diffusion of therapeutic agents through the blood clot result in the relatively low delivery efficiency into the deep wound site. Here, a microalgae-based biohybrid microrobot for accelerated diabetic wound healing is developed. The biohybrid microrobot autonomously moves at velocity of 33.3 µm s-1 and generates oxygen for the alleviation of hypoxic condition. In addition, the microrobot efficiently bound with inflammatory chemokines of interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) for modulating the immune responses. The enhanced penetration of microrobot is corroborated by measuring fibrin clots in biomimetic wound using microfluidic devices and the enhanced retention of microrobot is confirmed in the real wounded mouse skin tissue. After deposition on the chronic wound in diabetic mice without wound dressing, the wounds treated with microrobots are completely healed after 9 days with the significant decrease of inflammatory cytokines below 31% of the control level and the upregulated angiogenesis above 20 times of CD31+ cells. These results confirm the feasibility of microrobots as a next-generation platform for diabetic wound healing.

Manganese and cobalt recovery by surface display of metal binding peptide on various loops of OmpC in Escherichia coli.

In a cell-surface display (CSD) system, successful display of a protein or peptide is highly dependent on the anchoring motif and the position of the display in that anchoring motif. In this study, a recombinant bacterial CSD system for manganese (Mn) and cobalt (Co) recovery was developed by employing OmpC as an anchoring motif on three different external loops. A portion of Cap43 protein (TRSRSHTSEG)3 was employed as a manganese and cobalt binding peptide (MCBP), which was fused with OmpC at three different external loops. The fusions were made at the loop 2 [fusion protein-2 (FP2)], loop 6 (FP6), and loop 8 (FP8) of OmpC, respectively. The efficacy of the three recombinant strains in the recovery of Mn and Co was evaluated by varying the concentration of the respective metal. Molecular modeling studies showed that the short trimeric repeats of peptide probably form a secondary structure with OmpC, thereby giving rise to a difference in metal recovery among the three recombinant strains. Among the three recombinant strains, FP6 showed increased metal recovery with both Mn and Co, at 1235.14 (1 mM) and 379.68 (0.2 mM) µmol/g dry cell weight (DCW), respectively.

Effects of Sparassis crispa in Medical Therapeutics: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

In this study, we investigated the therapeutic potential and medical applications of Sparassis crispa (S. crispa) by conducting a systematic review of the existing literature and performing a meta-analysis. The original efficacy treatment of the mushroom extract is considered primarily and searched in electronic databases. A total of 623 articles were assessed, 33 randomized controlled experiments were included after the manual screening, and some papers, review articles, or editorials that did not contain data were excluded. A comparative standard means difference (SMD) and a funnel plot between control and S. crispa groups were used as parameters to demonstrate the beneficial effects of S. crispa for diabetes and cancer treatment, as well as anti-inflammatory, anti-fungal and antioxidant activities. The meta-analysis was carried out using Review Manager 5.1 software. Although for therapeutic diabetes there was heterogeneity in the subgroup analysis (I² = 91.9%), the overall results showed statistically significant SMDs in major symptoms that decreased serum insulin levels (SMD = 1.92, 95% CI (1.10, 2.75), I² = 0%), wound rates (SMD = 3.55 (2.56, 4.54), I² = 40%) and contributions to an increase in nutrient intake content (SMD = 0.32 (-0.15, 0.78), I² = 0%). Simultaneously, the study confirmed the utility of S. crispa treatment in terms of not only anti-cancer activity (reduction of tumor activity and survival of cancer cells I² = 42 and 34%, respectively) but also anti-inflammatory, anti-fungal and antioxidant activities (I² = 50, 44, and 10%, respectively). Our findings suggest that S. crispa extracts are useful for prevention and treatment of human diseases and might be the best candidates for future medicines.

Lithographically Encrypted Inverse Opals for Anti-Counterfeiting Applications.

Colloidal photonic crystals possess inimitable optical properties of iridescent structural colors and unique spectral shape, which render them useful for security materials. This work reports a novel method to encrypt graphical and spectral codes in polymeric inverse opals to provide advanced security. To accomplish this, this study prepares lithographically featured micropatterns on the top surface of hydrophobic inverse opals, which serve as shadow masks against the surface modification of air cavities to achieve hydrophilicity. The resultant inverse opals allow rapid infiltration of aqueous solution into the hydrophilic cavities while retaining air in the hydrophobic cavities. Therefore, the structural color of inverse opals is regioselectively red-shifted, disclosing the encrypted graphical codes. The decoded inverse opals also deliver unique reflectance spectral codes originated from two distinct regions. The combinatorial code composed of graphical and optical codes is revealed only when the aqueous solution agreed in advance is used for decoding. In addition, the encrypted inverse opals are chemically stable, providing invariant codes with high reproducibility. In addition, high mechanical stability enables the transfer of the films onto any surfaces. This novel encryption technology will provide a new opportunity in a wide range of security applications.

Microfluidic Production of Semipermeable Microcapsules by Polymerization-Induced Phase Separation.

Semipermeable microcapsules are appealing for controlled release of drugs, study of cell-to-cell communication, and isolation of enzymes or artificial catalysts. Here, we report a microfluidic strategy for creating monodisperse microcapsules with size-selective permeability using polymerization-induced phase separation. Monodisperse water-in-oil-in-water (W/O/W) double-emulsion drops, whose ultrathin middle layer is composed of photocurable resin and inert oil, are generated in a capillary microfluidic device, and irradiated by UV light. Upon UV illumination, the monomers are photopolymerized, which leads to phase separation between the polymerized resin and the oil within the ultrathin shell. Subsequent dissolution of the oil leaves behind regular pores in the polymerized membrane that interconnect the interior and exterior of the microcapsules, thereby providing size-selective permeability. The degree of phase separation can be further tuned by adjusting the fraction of oil in the shell or the affinity of the oil to the monomers, thereby enabling the control of the cutoff value of permeation. High mechanical stability and chemical resistance of the microcapsules, as well as controllable permeability and high encapsulation efficiency, will provide new opportunity in a wide range of applications.

Aquatic ecotoxicity effect of engineered aminoclay nanoparticles.

In the present study the short term aquatic ecotoxicity of water-solubilized aminoclay nanoparticles (ANPs) of ~51±31 nm average hydrodynamic diameter was characterized. An ecotoxicological evaluation was carried out utilizing standard test organisms of different phyla and trophic levels namely the eukaryotic microalga Pseudokirchneriella subcapitata, the crustacean Daphnia magna and the bioluminescent marine bacteria Vibrio fisheri. The effective inhibitory concentration (EC50) with 95% confidence limits for the microalga was 1.29 mg/L (0.72-1.82) for the average growth rate and 0.26 mg/L (0.23-0.31) for the cell yield. The entrapping of algal cells in aggregates of ANP may play a major role in the growth inhibition of algae P. subcapitata. No inhibition was observed for V. fisheri up to 25,000 mg/L (no observed effect concentration; NOEC). For D. magna no immobilization was observed in a limit test with 100 mg/L in 24 h while in 48 h a single animal was immobilized (5% inhibition). Correspondingly, the NOEC of ANP in 24 h was 100 mg/L and the lowest observed effect concentration (LOEC) for 48 h was 100 mg/L. Therefore it can be considered to use ANP as an algal-inhibition agent at concentrations <100 mg/L without affecting or only mildly affecting other organisms including zooplanktons, but further studies on the environmental fate and chronic toxicity of ANP is needed to confirm this.

Mixotrophic cultivation of oleaginous Chlorella sp. KR-1 mediated by actual coal-fired flue gas for biodiesel production.

Flue gases mainly consist of CO2 that can be utilized to facilitate microalgal culture for bioenergy production. In the present study, to evaluate the feasibility of the utilization of flue gas from a coal-burning power plant, an indigenous and high-CO2-tolerant oleaginous microalga, Chlorella sp. KR-1, was cultivated under mixotrophic conditions, and the results were evaluated. When the culture was mediated by flue gas, highest biomass (0.8 g cells/L·d) and FAME (fatty acid methyl esters) productivity (121 mg/L·d) were achieved in the mixotrophic mode with 5 g/L glucose, 5 mM nitrate, and a flow rate of 0.2 vvm. By contrast, the photoautotrophic cultivation resulted in a lower biomass (0.45 g cells/L·d) and a lower FAME productivity (60.2 mg/L·d). In general, the fatty acid profiles of Chlorella sp. KR-1 revealed meaningful contents (>40 % of saturated and mono-unsaturated fatty acids) under the mixotrophic condition, which enables the obtainment of a better quality of biodiesel than is possible under the autotrophic condition. Conclusively then, it was established that a microalgal culture mediated by flue gas can be improved by adoption of mixotrophic cultivation systems.

Development of dual strain microalgae cultivation system for the direct carbon dioxide utilization of power plant flue gas.

This study aims to propose a biological system that allows for direct utilization of flue gas for carbon dioxide capture and utilization by microalgae. The strain Chlorella sp. ABC-001 is employed for its high growth rate as well as lipid and carbohydrate content. Toxicity tests showed that cell growth was unaffected by NO, but the presence of SO2 showed critical damage on cell growth. Hence, an extremophile alga, Galdieria sulphuraria 5587.1 was applied to build a dual-strain cultivation system to mitigate the effect of SO2 toxicity and increase CO2 capture efficiency. All SO2 was removed by Galdieria culture and the system exhibited stable growth from a simulated flue gas stream containing CO2, NO and SO2. Combined CO2 biofixation rate of 793 mg/L/d and lipid productivity of 113 mg/L/d was achieved. The results showed that this new cultivation system is a promising alternative for reducing CO2 emissions from power plants.

Biomass, lipid content, and fatty acid composition of freshwater Chlamydomonas mexicana and Scenedesmus obliquus grown under salt stress.

Two freshwater microalgae including Chlamydomonas mexicana and Scenedesmus obliquus were grown on Bold Basal Medium (BBM) with different levels of salinity up to 100 mM NaCl. The dry biomass and lipid content of microalgae were improved as the concentration of NaCl increased from 0 to 25 mM. Highest dry weight (0.8 and 0.65 g/L) and lipid content (37 and 34 %) of C. mexicana and S. obliquus, respectively, were obtained in BBM amended with 25 mM NaCl. The fatty acid composition of the investigated species was also improved by the increased NaCl concentration. At 50 mM, NaCl palmitic acid (35 %) and linoleic acid (41 %) were the dominant fatty acids in C. mexicana, while oleic acid (41 %) and α-linolenic acid (20 %) were the major fractions found in S. obliquus.

Simultaneous enhancement of lipid biosynthesis and solvent extraction of Chlorella using aminoclay nanoparticles.

Magnesium aminoclay nanoparticles (MgANs) exert opposing effects on photosynthetic microalgae by promoting carbon dioxide (CO2) uptake and inducing oxidative stress. This study explored the potential application of MgAN in the production of algal lipids under high CO2 concentrations. The impact of MgAN (0.05-1.0 g/L) on cell growth, lipid accumulation, and solvent extractability varied among three tested oleaginous Chlorella strains (N113, KR-1, and M082). Among them, only KR-1 exhibited significant improvement in both total lipid content (379.4 mg/g cell) and hexane lipid extraction efficiency (54.5%) in the presence of MgAN compared to those of controls (320.3 mg/g cell and 46.1%, respectively). This improvement was attributed to the increased biosynthesis of triacylglycerols and a thinner cell wall based on thin-layer chromatography and electronic microscopy, respectively. These findings suggest that using MgAN with robust algal strains can enhance the efficiency of cost-intensive extraction processes while simultaneously increasing the algal lipid content.

Cell disruption and astaxanthin extraction from Haematococcus pluvialis: Recent advances.

The green microalga Haematococcus pluvialis is an excellent source of astaxanthin, a powerful antioxidant widely used in cosmetics, aquaculture, health foods, and pharmaceuticals. This review explores recent developments in cell disruption and astaxanthin extraction techniques applied using H. pluvialis as a model species for large-scale algal biorefinery. Notably, this alga develops a unique cyst-like cell with a rigid three-layered cell wall during astaxanthin accumulation (∼4% of dry weight) under stress. The thick (∼2 µm), acetolysis-resistant cell wall forms the strongest barrier to astaxanthin extraction. Various physical, chemical, and biological cell disruption methods were discussed and compared based on theoretical mechanisms, biomass status (wet, dry, and live), cell-disruption efficacy, astaxanthin extractability, cost, scalability, synergistic combinations, and impact on the stress-sensitive astaxanthin content. The challenges and future prospects of the downstream processes for the sustainable and economic development of advanced H. pluvialis biorefineries are also outlined.

Cell disruption and lipid extraction from Chlorella species for biorefinery applications: Recent advances.

Chlorella is a promising microalga for CO2-neutral biorefinery that co-produces drop-in biofuels and multiple biochemicals. Cell disruption and selective lipid extraction steps are major technical bottlenecks in biorefinement because of the inherent robustness and complexity of algal cell walls. This review focuses on the state-of-the-art achievements in cell disruption and lipid extraction methods for Chlorella species within the last five years. Various chemical, physical, and biological approaches have been detailed theoretically, compared, and discussed in terms of the degree of cell wall disruption, lipid extractability, chemical toxicity, cost-effectiveness, energy use, scalability, customer preferences, environment friendliness, and synergistic combinations of different methods. Future challenges and prospects of environmental-friendly and efficient extraction technologies are also outlined for practical applications in sustainable Chlorella biorefineries. Given the diverse industrial applications of Chlorella, this review may provide useful information for downstream processing of the advanced biorefineries of other algae genera.

Rapid estimation of triacylglycerol content of Chlorella sp. by thermogravimetric analysis.

A simple and reliable method based on thermogravimetric analysis has been developed for determining triacylglycerol content in Chlorella sp. KR-1. There are two decomposing steps during pyrolysis of the microalgal cells and the second step of weight loss may be attributed to degradation and volatilization of triacylglycerols. The second peak height in the temperature derivatives of weight loss increased with the triacylglycerol content of the microalgal cells and the peak was around 390 °C regardless of the triacylglycerol contents. Based on these findings, a linear equation for determining triacylglycerol content was derived. The proposed method gives satisfactory results, showing small variance and a good interpolation capability.

Correlation of microbial community with salinity and nitrogen removal in an anammox-based denitrification system.

Anaerobic ammonium oxidation (anammox), a low-energy-consuming technology, can be used to remove nitrogen from industrial saline wastewater. However, high salinity inhibits anammox microbial activity. This study investigated the effect of salinity on nitrogen removal performance and microbial community structure. The experiment used an up-flow anammox reactor fed with synthetic wastewater with salinity increased from 0.5 to 2.5%. Results indicated that 80% nitrogen removal efficiency can be achieved at 2% salinity with a nitrogen loading rate of 2.0 kg-N/m3/d. Anammox performance significantly deteriorated at 2.5% salinity. High-throughput sequencing revealed that Planctomycetes (representative anammox bacteria) increased with salinity, replacing Proteobacteria (representative heterotrophic denitrifying bacteria) in the microbial community. qPCR analysis indicated that relative abundance of "Candidatus Kuenenia" within anammox bacteria increased from 3.96 to 83.41%, corresponding to salinity of 0.5-2.0%, and subsequently decreased to 63.27% at 2.5% salinity, correlating with nitrogen-removal performance. Thus, anammox has potential in nitrogen removal from wastewater with salinity up to 2%.

Enabling anoxic acetate assimilation by electrode-driven respiration in the obligate aerobe, Pseudomonas putida.

This study examined the obligate aerobe, Pseudomonas putida, using acetate as the sole carbon and energy source, and respiration via an anode as the terminal electron acceptor under anoxic conditions. P. putida showed significantly different acetate assimilation in a closed-circuit microbial fuel cell (CC-MFC) compared to an open circuit MFC (OC-MFC). More than 72% (2.6 mmol) of acetate was consumed during 84 hrs in the CC-MFC in contrast to the no acetate consumption observed in the OC-MFC. The CC-MFC produced 150 µA (87 C) from acetate metabolization. Electrode-based respiration reduced the NADH/NAD+ ratio anaerobically, which is similar to the aerobic condition. The CC-MFC showed significantly higher acetyl-CoA synthetase activity than the OC-MFC (0.028 vs. 0.001 µmol/min/mg), which was comparable to the aerobic condition (circa 60%). Overall, electrode-based respiration enables P. putida to metabolize acetate under anoxic conditions and provide a platform to regulate the bacterial redox balance without oxygen.

Photoautotrophic hydrogen production of Rhodobacter sphaeroides in a microbial electrosynthesis cell.

Conventional photoheterotrophic H2 production by purple sulfur bacteria requires additional organic substrates as the carbon and energy sources. This study examined the novel photoautotrophic H2 production of Rhodobacter sphaeroides with concomitant CO2 uptake in microbial electrosynthesis (MES). Under an applied potential of -0.9 V vs. Ag/AgCl to the cathode, Rhodobacter sphaeroides produced hydrogen with CO2 as the sole carbon source under illumination. The initial planktonic cells decreased rapidly in suspension, whereas biomass formation on the cathode surface increased gradually during MES operation. The electron and carbon flow under photoautotrophic conditions in MES were estimated. Glutamate, as the nitrogen source, enhanced hydrogen production significantly (328 mL/L/day) compared to NH4Cl (67 mL/L/day) during seven days of operation. The photoautotrophic condition with 6000 lx presented CO2 consumption and simultaneous biomass formation on the cathode electrode. MES-driven electron and proton transfer enabled the simultaneous production of hydrogen and CO2 uptake.

Recent developments and key barriers to microbial CO2 electrobiorefinery.

The electrochemical conversion of CO2 can include renewable surplus electricity storage and CO2 utilisation. This review focuses on the microbial CO2 electrobiorefinery based on microbial electrosynthesis (MES) which merges electrochemical and microbial conversion to produce biofuels and higher-value chemicals. In this review, recent developments are discussed about bioelectrochemical conversion of CO2 into biofuels and chemicals in MES via microbial CO2-fixation and electricity utilisation reactions. In addition, this review examines technical approaches to overcome the current limitations of MES including the following: engineering of the biocathode, application of electron mediators, and reactor optimisation, among others. An in-depth discussion of strategies for the CO2 electrobiorefinery is presented, including the integration of the biocathode with inorganic catalysts, screening of novel electroactive microorganisms, and metabolic engineering to improve target productivity from CO2.

Supply of proton enhances CO electrosynthesis for acetate and volatile fatty acid productions.

The microbial electrosynthesis is a platform to supply protons and electrons to improve the conversion efficiency and production rate for the valorization of C1 gas. This study examined proton migration and electron transfer of the electrode and microbe by using various external parameters in the electrosynthesis of CO. The CO electrosynthesis achieved almost double of coulombic efficiency than the conventional CO2 electrosynthesis. The maximum volumetric acetate production rate was 0.71 g/L/day in the BES, which was 2-6 times higher than reported elsewhere. These results show that the efficient proton migration and electron transfer can enhance the productivity and conversion efficiency of the biological CO conversion in a bioelectrochemical system.

Effects of Nitrogen Supplementation Status on CO2 Biofixation and Biofuel Production of the Promising Microalga Chlorella sp. ABC-001.

The use of microalgal biomass as feedstock for biofuels has been discussed for decades as it provides a sustainable approach to producing fuels for the future. Nonetheless, its feasibility has not been established yet and various aspects of biomass applications such as CO2 biofixation should also be explored. Therefore, in this study, the CO2 biofixation and lipid/carbohydrate production potential of Chlorella sp. ABC-001 were examined under various nitrogen concentrations. The highest biomass productivity and CO2 biofixation rate of 0.422 g/l/d and 0.683 g/l/d, respectively, were achieved under a nitrogen-rich condition (15 mM nitrate). Carbohydrate content was generally proportional to initial nitrate concentration and showed the highest value of 41.5% with 15 mM. However, lipid content showed an inverse relationship with nitrogen supplementation and showed the highest value of 47.4% with 2.5 mM. In consideration as feedstock for biofuels (bioethanol, biodiesel, and biogas), the sum of carbohydrate and lipid contents were examined and the highest value of 79.6% was achieved under low nitrogen condition (2.5 mM). For lipid-based biofuel production, low nitrogen supplementation should be pursued. However, considering the lower feasibility of biodiesel, pursuing CO2 biofixation and the production of carbohydrate-based fuels under nitrogenrich condition might be more rational. Thus, nitrogen status as a cultivation strategy must be optimized according to the objective, and this was confirmed with the promising alga Chlorella sp. ABC-001.

Recovery of Astaxanthin-Containing Oil from Haematococcus pluvialis by Nano-dispersion and Oil Partitioning.

The feasibilities of cell disruption by homogenization-assisted high-pressure nano-dispersion and recovery of astaxanthin-containing oil by oil partitioning in oil-acetone-water solution were examined. The total fatty acid content of Haematococcus pluvialis was 414.6 mg/g cell, and the astaxanthin content was 4.4% of oil. Extra oil was added to the solution in order to recover oil through instability of dispersion status instead of solvent evaporation. A total amount of energy of 0.34 kWh/L was consumed for acetone evaporation at 50 °C, whereas fully 1.86 kWh/L of energy for water evaporation was consumed. When soybean oil was added to the solution after partial acetone evaporation, the oil-recovery yield was 97.8%, while the yield after full evaporation was 97.6% in 10-g/L solution. However, the energy consumed for partial evaporation (0.29 kWh/L) was much lower than that for full evaporation (0.40 kWh/L). When H. pluvialis oil was added to the solution after partial evaporation, the oil-recovery yield decreased to 90.6% due to the impurity of crude H. pluvialis oil in 10-g/L solution. Methods such as refining of H. pluvialis oil, increase of microalgae dosage for cell disruption, and increase of the injection amount of extra oil can help to enhance oil recovery.