Multiplex PCR system to track authorized and unauthorized genetically modified soybean events in food and feed

A diverse range of genetic elements has been used to develop genetically modified organisms (GMOs) over the last 18 years. Screening methods that target few elements, such as the Cauliflower Mosaic Virus 35S promoter (P-35S) and Agrobacterium tumefaciens nopaline terminator (T-nos), are not sufficient to screen GMOs. In the present study, a multiplex PCR system for all globally commercialized GM soybean events was developed to easily trace the events. For this purpose, screening elements of 24 GM soybean events were investigated and 9 screening targets were selected and divided into three individual triplex PCR systems: P-35S, ribulose-1,5-bisphosphate carboxylase small subunit promoter of Arabidopsis thaliana, T-nos, T-35S, pea E9 terminator, open reading frame 23 terminator of A. tumefaciens, proteinase inhibitor II terminator of potato, acetohydroxy acid synthase large subunit terminator of A. thaliana, and the revealed 3′ flanking sequences of DP-305423-1. The specificity of the assays was confirmed using thirteen GM soybean events as the respective positive/negative controls. The limit of detection of each multiplex set, as determined using certified reference materials of specific GM events, ranged from 0.03 to 0.5%, depending upon target. Furthermore, 26 food samples that contained soybean ingredients, which were purchased from the USA, China, Japan, and Korea, were analyzed, 17 of which contained one or more GM soybean events. These results suggest that the developed screening method can be used to efficiently track and identify 24 GM soybean events in food and feed.     

Acquisition of amino acids by Lactobacillus delbrueckii subsp. bulgaricus 2038 when grown in the presence of casein

The acquisition of amino acids by Lactobacillus delbrueckii subsp. bulgaricus 2038 (Lb. bulgaricus 2038) when grown in the presence of bovine casein, the major protein in bovine milk, was investigated by examining the expression of genes related to proteolysis and amino acid biosynthesis. To support the growth on bovine casein, Lb. bulgaricus 2038 has to synthesise five kinds of amino acids de novo, as proteolysis from casein does not provide these. The incomplete hydrolysis in combination with amino acids biosynthesis may explain the slow growth of Lb. bulgaricus 2038 in a casein environment. Meanwhile, it was determined that Lb. bulgaricus 2038 uses different intracellular peptidases when grown in casein or whey medium, and initially yields the important amino acid glutamate from the C-terminal or N-terminal end of peptides imported into the cell.     

Improvement of sediment microbial fuel cell performances by design and application of power management systems

One of the ways to generate clean and non-destructive energy is to use the energy stored in the biomass resources by the microbial fuel cells (MFCs). Sediment Microbial Fuel Cells (SMFCs) are a special type of MFCs that use organic materials in aquifers sediment to generate electricity. In this research, the effects of an increase in the electrode surface are investigated. The results showed that the increase in cathode electrode surface had better efficiency than the multi-cathode mode (maximum power generated for a 3-cathode electrode (27 cm³) and 1-cathode electrode (27 cm³) was 526 mW/cm² and 800 mW/cm², respectively. Another parameter affecting the performance of these systems is temperature. In the next step, the power generation rate was measured in different step currents and at different sample times. In the final stage, a power management system (PMS) was designed to optimally utilize the output energy of the improved SMFC, leading to an increase in the output voltage to 3.3 V.     

Improvement in energy recovery from Chlorella sp. biomass by integrated dark-photo biohydrogen production and dark fermentation-anaerobic digestion processes

Chlorella sp. biomass was used as the sole substrate for the production of hydrogen and methane through integrated dark fermentation (DF) and photo-fermentation (PF), and DF and anaerobic digestion (AD) processes. Prior to use in fermentations, the biomass was pretreated by acid-hydrothermal method, which yielded a maximum reducing sugar yield of 162.9 ± 4.2 mg g-biomass⁻¹. The use of the microalgal hydrolysate to produce hydrogen by DF gave a hydrogen yield (HY) of 47.2 ± 1.1 mL g-volatile-solids⁻¹ (VS). The subsequent use of the hydrogenic effluent in PF gave a HY of 125.0 ± 1.5 mL g-VS⁻¹, while AD of the hydrogenic effluent gave a methane yield of 152.8 ± 1.3 mL g-VS⁻¹. The total energy yield attained by the use of DF alone, the integrated DF-PF, and DF-AD processes were 0.51, 1.86 and 5.98 kJ g-VS⁻¹, respectively. These results indicate that the integrated DF-AD process was effective in recovering energy from Chlorella sp. biomass. However, an energy balance analysis indicated that the process was not energetically feasible due to the high energy demand for the acid-hydrothermal pretreatment.     

Improvement of biohythane production from Chlorella sp. TISTR 8411 biomass by co-digestion with organic wastes in a two-stage fermentation

The suitability of molasses, Napier grass (Pennisetum purpureum), empty fruit bunches (EFB), palm oil mill effluent (POME), and glycerol waste as a co-substrate with Chlorella sp. TISTR 8411 biomass for biohythane production was investigated. Mono-digestion of Chlorella biomass had hydrogen and methane yield of 23–35 and 164–177 mL gVS⁻¹, respectively. Co-digestion of Chlorella biomass with 2–6% TS of organic wastes was optimized for biohythane production with hydrogen and methane yield of 17–75 and 214–577 mL gVS⁻¹, respectively. The hydrogen and methane yield from co-digestion of Chlorella biomass with molasses, POME, and glycerol waste was increased by 8–100% and 80–264%, respectively. The biohythane production of co-digestion of Chlorella was 6–11 L L-mixed waste⁻¹ with an optimal C/N ratio range of 19–41 and H₂/CH₄ ratio range of 0.06–0.3. Co-digestion of Chlorella biomass was significantly improved biohythane production in term of yield, production rate, and kinetics.     

Enhanced bioactivity of Zanthoxylum schinifolium fermented extract: Anti-inflammatory,anti-bacterial,and anti-melanogenic activity

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Effect of substituents of twisted benzodiperylenediimides on non-fullerene solar cells

Twisted benzodiperylenediimides (TBDPDI) with large rigid conjugated core and strong absorption is regarded as an excellent acceptor in non-fullerene solar cells. Since side chains of semiconductors play a crucial role in the solar cells, TBDPDI acceptors with different side chains (1-ethylpropyl, C5; 2-ethylhexyl, C8; 1-pentylhexyl, C11; 2-octyldodecyl, C20; 1-undecyldodecyl, C23) were synthesized. In solution, TBDPDI compounds (C5, C11, and C23) with alkyl chains branched at 1-position show significantly different absorption profiles and fluorescence intensity with those (C8 and C20) branched at 2-position, due to stronger aggregation of the latter. Nevertheless, alkyl chains have little effect on the molecular orbital energy levels and optical band gaps, as verified by cyclic voltammetry and solid state absorption. Due to their complementary absorption and matchable energy levels with donor of PCE10, these acceptors and PCE10 were used together to fabricate bulk heterojunction (BHJ) solar cells. Because of inferior phase separation with large domain size around 100 nm and bulky insulated side chains, acceptors (C20 and C23) with long alkyl chains have the low electron mobility (μ_e) around 10⁻⁸ cm² V⁻¹ s⁻¹ and the low power conversion efficiency (PCE) of solar cells. TBDPDI (C11) with 1-pentylhexyl gives the highest PCE of 5.0% under the optimized condition, which is attributed to proper phase separation with domain size around 20 nm and highest μ_e of 10⁻⁶ cm² V⁻¹ s⁻¹.     

Surface coating with poly(trifluoroethyl methacrylate) through rapid expansion of supercritical CO2 solutions

Rapid expansion of supercritical solutions (RESS) of poly(trifluoroethyl methacrylate), poly(TFEMA), was performed to produce ultrafine particles for spray coating application to improve the hydrophobicity of moisture-sensitive biodegradable materials. Carbon dioxide (CO₂) was used as the RESS solvent. Thermoplastic starch/poly(butylene adipate-co-terephthalate) (TPS/PBAT, 60:40 wt/wt) blend was used as the coating substrate. The objectives of this work were to determine the capacity of the RESS process for coating TPS-based material with poly(TFEMA), and to investigate the effect of RESS parameters – i.e. pre-expansion pressure and temperature (P_pre, T_pre) and poly(TFEMA) concentration – on the surface morphology and hydrophobicity of the coated materials. It was found that RESS produced poly(TFEMA) particles precipitated onto the surface of the TPS/PBAT substrate, with particle sizes ranging from 30 nm to several microns, depending on processing parameters. Rapid expansion of fluoropolymer solutions (0.3–1.0 wt%) with P_pre of 331 bar initiated from unsaturated conditions produced nanoparticles with a narrow size distribution of ∼30–70 nm; whereas larger particles with broader size distributions and a lower degree of agglomeration were obtained when supersaturated solutions were expanded with P_pre of 172 bar, especially at T_pre (80 °C) – higher than the glass transition temperature (73 °C) of poly(TFEMA). The surface coverage by the fluoropolymer increased with increasing P_pre and poly(TFEMA) concentration, but decreased with increasing T_pre. In addition, the hydrophobicity of the coated substrate, determined by water contact angle and water vapor transmission rate measurements, increased with increasing surface coverage.     

Vegetable waste as substrate and source of suitable microflora for bio-hydrogen production

Self-fermentation of cellulosic substrates to produce biohydrogen without inoculum addition nor pretreatments was investigated. Dark fermentation of two different substrates made of leaf-shaped vegetable refuses (V) and leaf-shaped vegetable refuses plus potato peels (VP), was taken in consideration. Batch experiments were carried out, under two mesophilic anaerobic conditions (28 and 37 °C), in order to isolate and to identify potential H₂-producing bacterial strains contained in the vegetable extracts. The effect of initial glucose concentration (at 1, 5 and 10 g/L) on fermentative H₂ production by the isolates was also evaluated.H₂ production from self-fermentation of both biomasses was found to be feasible, without methane evolution, showing the highest yield for V biomass at 28 °C (24 L/kg VS). The pH control of the culture medium proved to be a critical parameter. The isolates had sequence similarities ≥98% with already known strains, belonging to the family Enterobacteriaceae (γ-proteobacteria) and Streptococcaceae (Firmicutes). Four genera found in the samples, namely Pectobacterium, Raoultella, Rahnella and Lactococcus have not been previously described for H₂ production from glucose. The isolates showed higher yield (1.6–2.2 mol H₂/mol glucose_added) at low glucose concentration (1 g/L), while the maximum H₂ production ranged from 410 to 1016 mL/L and was obtained at a substrate concentration of 10 g/L. The results suggested that vegetable waste can be effectively used as both, substrate and source of suitable microflora for bio-hydrogen production.