Enhanced ophthalmic bioavailability and stability of atropine sulfate via sustained release particles using polystyrene sulfonate resin.

Atropine sulfate (ATS) eye drops at low concentrations constitute a limited selection for myopia treatment, with challenges such as low ophthalmic bioavailability and inadequate stability. This study proposes a novel strategy by synthesizing ophthalmic sodium polystyrene sulfonate resin (SPSR) characterized by a spherical shape and uniform size for cationic exchange with ATS. The formulation of ATS@SPSR suspension eye drops incorporates xanthan gum and hydroxypropyl methylcellulose (HPMC) as suspending agents. In vitro studies demonstrated that ATS@SPSR suspension eye drops exhibited sustained release characteristics, and tropic acid, its degradation product, remained undetected for 30 days at 40 °C. The ATS levels in the tear fluids and aqueous humor of New Zealand rabbits indicated a significant increase in mean residence time (MRT) and area under the drug concentration-time curve (AUC0-12h) for ATS@SPSR suspension eye drops compared to conventional ATS eye drops. Moreover, safety assessment confirmed the non-irritating nature of ATS@SPSR suspension eye drops in rabbit eyes. In conclusion, the cation-responsive sustained-release ATS@SPSR suspension eye drops enhanced the bioavailability and stability of ATS, offering a promising avenue for myopia treatment.

Controlled release of vitamin A palmitate from crosslinked cyclodextrin organic framework for dry eye disease therapy.

Controlled release drug delivery systems of eye drops are a promising ophthalmic therapy with advantages of good patient compliance and low irritation. However, the lack of a suitable drug carrier for ophthalmic use limits the development of the aforementioned system. Herein, the crosslinked cyclodextrin organic framework (COF) with a cubic porous structure and a uniform particle size was synthesized and applied to solidify vitamin A palmitate (VAP) by using the solvent-free method. The VAP@COF suspension eye drops were formulated by screening co-solvents, suspending agents, and stabilizing agents to achieve a homogeneous state and improve stability. According to the in vitro release study, the VAP@COF suspension exhibited a controlled release of VAP within 12 h. Both the ex vivo corneal contact angle and in vivo fluorescence tracking indicated that the VAP@COF suspension prolonged the VAP residence time on the ocular surface. This suspension accelerated the recovery of the dry eye disease (DED) model in New Zealand rabbits. Furthermore, the suspension was non-cytotoxic to human corneal epithelial cells and non-irritation to rabbit eyes. In summary, the particulate COF is an eye-acceptable novel carrier that sustains release and prolongs the VAP residence time on the ocular surface for DED treatment.

Reverse design of haptens based on antigen spatial conformation to prepare anti-capsaicinoids&gingerols antibodies for monitoring of gutter cooking oil.

Rapid simultaneous detection of multi-component adulteration markers can improve the accuracy of identification of gutter cooking oil in edible oil, which is made possible by broad-spectrum antibody (bs-mAb). This study used capsaicinoids (CPCs) and gingerol derivatives (GDs) as adulteration markers, and two broad-spectrum haptens (bs-haptens) were designed and synthesized based on a reverse design strategy of molecular docking. Electrostatic potential (ESP) and monoclonal antibodies (mAbs) preparation verified the strategy's feasibility. To further investigate the recognition mechanism, five other reported antigens and mAbs were also used. Finally, the optimal combination (Hapten 5-OVA/1-F12) and key functional groups (f-groups) were determined. The half maximal inhibitory concentration (IC50) for CPCs-GDs was between 88.13 and 499.16 ng/mL. Meanwhile, a preliminary lateral flow immunoassay (LFIA) study made practical monitoring possible. The study provided a theoretical basis for the virtual screening of bs-haptens and simultaneous immunoassay of multiple exogenous markers to monitor gutter oil rapidly and accurately.

Silver/copper bimetallic nanoclusters integrating with cryonase-assisted target recycling amplification detection of Salmonella typhimurium.

An unsophisticated fluorescence-enabled strategy is brought forward to process the highly sensitive fluorescence detection of Salmonella typhimurium (S. typhimurium) which based on polyethyleneimine (PEI)-templated silver/copper nanoclusters (Ag/CuNCs) (λ excitation = 334 nm and λ emission = 466 nm) with cryonase-assisted target recycling amplification. The Ag/CuNCs nanoclusters are synthesized as fluorescent materials due to their strong and stable fluorescence characteristics and are modified with S. typhimurium aptamers to form aptamer-Ag/CuNCs probes. The probes can be adsorbed on the surface of quenching agents-polydopamine nanospheres (PDANSs), thereby inducing fluorescence quenching of the probes. Once the aptamers are bound to the target, the aptamers/targets complexes are separated from the PDANSs surface, and the Ag/CuNCs recover the fluorescence signal. The released complexes will immediately be transformed into a substrate digested by cryonase (an enzyme that can digest all types of nucleic acids), and the released targets are bound to another aptamers to initiate the next round of cleavage. This reaction will be repeated continuously until all relevant aptamers are consumed and all Ag/CuNCs are released, resulting in a significant amplification of the fluorescence signal and improved sensitivity. Using Ag/CuNCs as fluorescent probes combined with cryonase-assisted amplification strategy, the fluorescence aptasensor is constructed with detection limits as low as 3.8 CFU mL-1, which is tenfold better than without the cryonase assistance. The method developed has been applied to milk, orange juice, chicken, and egg white samples with excellent selectivity and accuracy providing an approach for the early and rapid detection of S. typhimurium in food.

Paper-based biosensors relying on core biological immune scaffolds for the detection of procymidone in vegetables.

In order to achieve a highly sensitive detection of procymidone in vegetables, three paper-based biosensors based on a core biological immune scaffold (CBIS) were developed, which were time-resolved fluorescence immunochromatography strips with Europium (III) oxide (Eu-TRFICS). Goat anti-mouse IgG and europium oxide time-resolved fluorescent microspheres formed secondary fluorescent probes. CBIS was formed by secondary fluorescent probes and procymidone monoclonal antibody (PCM-Ab). The first type of Eu-TRFICS (Eu-TRFICS-(1)) fixed secondary fluorescent probes on a conjugate pad, and PCM-Ab was mixed with a sample solution. The second type of Eu-TRFICS (Eu-TRFICS-(2)) fixed CBIS on the conjugate pad. The third type of Eu-TRFICS (Eu-TRFICS-(3)) was directly mixed CBIS with the sample solution. They solved the problems of steric hindrance of antibody labeling, insufficient exposure of antigen recognition region and easy loss of activity in traditional methods. They realized multi-dimensional labeling and directional coupling. They replaced the loss of antibody activity. And the three types of Eu-TRFICS were compared, among which Eu-TRFICS-(1) was the best detection choice. Antibody usage was reduced by 25% and sensitivity was increased by 3 times. Its detection range was 1-800 ng/mL, the limit of detection (LOD) was 0.12 ng/mL with the visible LOD (vLOD) of 5 ng/mL.

Label-free colorimetric apta-assay for detection of Escherichia coli based on gold nanoparticles with peroxidase-like amplification.

In this work, aptamers against E. coli with better performance were obtained via cell systematic evolution of ligands by exponential enrichment (cell-SELEX) and dissociation constants (Kd) of aptamers were estimated to range from 133.87 to 199.44 nM. Furthermore, the selected aptamer was employed for label-free colorimetric detection of E. coli using gold nanoparticles (AuNPs) with peroxidase-like activity to catalyze the oxidation of tetramethylbenzidine (TMB) by hydrogen peroxide (H2O2) to produce color development. This colorimetric apta-assay started with an aptamer-bacteria binding step, and the concentration of residual aptamers after binding depended on the amount of target bacteria. Then, the amount of separated residual aptamers determined the degree of cetyltrimethylammonium bromide (CTAB)-inhibited catalytic activity of AuNPs, which resulted in a color change from dark blue to light blue. Owing to the excellent peroxidase activity of AuNPs, they could emit strong visible color intensity in less than 1 minute to improve visual detection sensitivity. Under optimized conditions, the sensitivity of detection was 5 × 103 CFU mL-1 visually and 75 CFU mL-1 using the UV-vis spectrum with a linear range from 5 × 102 to 1 × 106 CFU mL-1. And it had shown a good recovery rate in real samples of water, juice and milk compared with classical counting methods.

Novel Dual-Signal SiO2-COOH@MIPs Electrochemical Sensor for Highly Sensitive Detection of Chloramphenicol in Milk.

In view of the great threat of chloramphenicol (CAP) to human health and the fact that a few producers have illegally used CAP in the food production process to seek economic benefits in disregard of laws and regulations and consumer health, we urgently need a detection method with convenient operation, rapid response, and high sensitivity capabilities to detect CAP in food to ensure people's health. Herein, a molecularly imprinted polymer (MIP) electrochemical sensor based on a dual-signal strategy was designed for the highly sensitive analysis of CAP in milk. The NiFe Prussian blue analog (NiFe-PBA) and SnS2 nanoflowers were modified successively on the electrode surface to obtain dual signals from [Fe(CN)6]3-/4- at 0.2 V and NiFe-PBA at 0.5 V. SiO2-COOH@MIPs that could specifically recognize CAP were synthesized via thermal polymerization using carboxylated silica microspheres (SiO2-COOH) as carriers. When the CAP was adsorbed by SiO2-COOH@MIPs, the above two oxidation peak currents decreased at the same time, allowing the double-signal analysis. The SiO2-COOH@MIPs/SnS2/NiFe-PBA/GCE sensor used for determining CAP was successfully prepared. The sensor utilized the interactions of various nanomaterials to achieve high-sensitivity dual-signal detection, which had certain innovative significance. At the same time, the MIPs were synthesized using a surface molecular imprinting technology, which could omit the time of polymerization and elution and met the requirements for rapid detection. After optimizing the experimental conditions, the detection range of the sensor was 10-8 g/L-10-2 g/L and the limit of detection reached 3.3 × 10-9 g/L (S/N = 3). The sensor had satisfactory specificity, reproducibility, and stability, and was successfully applied to the detection of real milk samples.

Isolation of Bacteria Aptamers with Non-SELEX for the Development of a Highly Sensitive Colorimetric Assay Based on Dual Signal Amplification.

In this work, an aptamer against Escherichia coli is isolated via non-SELEX, which executes efficient selection by employing repetitive cycles of centrifugation-based partitioning, and the binding site of the aptamer on E. coli cell surfaces is inferred to be a membrane protein. Moreover, truncated sequence 2-17-2 with a higher affinity (Kd = 101.76 nM) is employed for highly sensitive colorimetric detection of bacteria based on the dual signal amplification strategy. When targets exist, the release of DNA 1 from the polymer activates a hybridization chain reaction (HCR) between DNA 1 and DNA 2, thereby inducing the aggregation of probe 1. Subsequently, DNA 3 dissociated from probe 1 as a linker DNA further assembles probe 2/3. In this system, two types of DNA@gold nanoparticles (AuNPs) coexist and successively aggregate AuNPs based on divergent triggering mechanisms. Under optimal conditions, the dual signal amplification strategy presents excellent sensitivity (10 CFU mL-1) and specificity, as well as the realization of real sample analysis.

Dual-target electrochemical DNA sensor for detection of Pb2+ and Hg2+ simultaneously by exonuclease I-assisted recycling signal amplification.

With the development of exonuclease, the exonuclease has been used to construct a variety of aptasensor and to realize the signal amplification. Among them, based on silver nanoparticles (Ag NPs) and exonuclease I (Exo I)-assisted cycle signal amplification strategy, we designed a novel high-sensitivity dual-target electrochemical biosensor to detect Pb2+ or Hg2+ in water. In the presence of Hg2+, the Hg2+ was fixed to the aptamer chain by thymine-Hg2+-thymine (T-Hg2+-T), resulting in the decrease of signal. When Pb2+ was present, DNA single strand S2 dissociated and was bound to Pb2+, which automatically triggered Exo I to selectively cut the single chain from the recognition site to achieve the cyclic amplification of the electrochemical signal. The interaction between aptamer and Exo I was investigated by gel electrophoresis. Under the optimum conditions in the scan range -0.20 to 0.60 V, the biosensor had high sensitivity with a linear range of 100 pg/L to 10.0 mg/L, Pb2+ or Hg2+, and the detection limits were 17.0 pg/L (R2 = 0.993) and 12.0 pg/L (R2 = 0.993), respectively. The relative standard deviation (RSD) of the sensor was 0.5-2.6%, and the recovery of spiked standard solutions was between 98.3 and 110%. The cycle amplification strategy supported by this enzyme has promising applications in detection of the two metal ions in various fields.

The Effects of Different Antigen-Antibody Pairs on the Results of 20 Min ELISA and 8 Min Chromatographic Paper Test for Quantitative Detection of Acetamiprid in Vegetables.

To establish rapid, high-sensitive, quantitative detection of ACP residues in vegetables. A 1G2 cell clone was selected as the most sensitive for anti-ACP antibody production following secondary immunization, cell fusion, and screening. The affinity of the 1G2 antibody to each of the four coating agents (imidacloprid−bovine serum albumin [BSA], thiacloprid−BSA, imidaclothiz−BSA, and ACP-BSA) was determined using a 20 min enzyme-linked immunosorbent assay (ELISA). The half maximal inhibitory concentration (IC50) was 0.51−0.62 ng/mL, showing no significant difference in affinity to different antigens. However, we obtained IC50 values of 0.58 and 1.40 ng/mL on the linear regression lines for 1G2 anti-ACP antibody/imidacloprid−BSA and 1G2 anti-ACP antibody/thiacloprid−BSA, respectively, via quantum dot (QD)-based immunochromatography. That is, the 1G2 antibody/imidacloprid−BSA pair (the best combination) was about three times more sensitive than the 1G2 antibody/thiacloprid−BSA pair in immunochromatographic detection. The best combination was used for the development of an 8 min chromatographic paper test. With simple and convenient sample pretreatment, we achieved an average recovery of 75−117%. The coefficient of variation (CoV) was <25% for all concentrations tested, the false−positive rate was <5%, the false−negative rate was 0%, and the linear range of the method was 50−1800 µg/kg. These performance metrics met the ACP detection standards in China, the European Union (EU), and the United States (US). In summary, in this study, we established an 8 min QD-based immunochromatographic stripe for the rapid and accurate quantitative determination of ACP residues in vegetables.

Electrochemiluminescence biosensor for determination of lead(II) ions using signal amplification by Au@SiO2 and tripropylamine-endonuclease assisted cycling process.

MXene@Au as the base and Au@SiO2 as signal amplification factor were used for constructing an ultrasensitive "on-off" electrochemiluminescence (ECL) biosensor for the detection of Pb2+ in water. The use of MXene@Au composite provided a good interface environment for the loading of tris(2,2-bipyridyl)ruthenium(II) (Ru(bpy)32+) on the electrode. Based on resonance energy transfer, the Au (core) SiO2 (shell) (Au@SiO2) nanoparticles stimulate electron transport and promote tripropylamine (TPrA) oxidation. The luminescence effect of Au@SiO2 was five times that of AuNPs and SiO2 nanomaterials alone, and the ECL intensity was greatly improved. In addition, Pb2+ activated the aptamer to exert its endonuclease activity, which realized the signal cycle amplification in the process of Pb2+ detection. When Pb2+ was added, the ECL signal weakened, and the Pb2+ concentration was detected according to the decreased ECL intensity. Under optimized experimental conditions, this aptamer sensor for Pb2+ has a wide detection range (0.1 to 1 × 106 ng L-1) and a low detection limit (0.059 ng L-1). The relative standard deviation (RSD) of the sensor is 0.39-0.99%, and the recovery of spiked standard is between 90.00 and 125.70%. The sensor shows good selectivity and high sensitivity in actual water sample analysis. This signal amplification strategy possibly provides a new method for the detection of other heavy metal ions and small molecules.

Novel Electrochemical Aptasensor Based on Ordered Mesoporous Carbon/2D Ti3C2 MXene as Nanocarrier for Simultaneous Detection of Aminoglycoside Antibiotics in Milk.

Herein, a novel electrochemical aptasensor using a broad-spectrum aptamer as a biorecognition element was constructed based on a screen-printed carbon electrode (SPCE) for simultaneous detection of aminoglycoside antibiotics (AAs). The ordered mesoporous carbon (OMC) was firstly modified on 2D Ti3C2 MXene. The addition of OMC not only effectively improved the stability of the aptasensor, but also prevented the stacking of Ti3C2 sheets, which formed a good current passage for signal amplification. The prepared OMC@Ti3C2 MXene functioned as a nanocarrier to accommodate considerable aptamers. In the presence of AAs, the transport of electron charge on SPCE surface was influenced by the bio-chemical reactions of the aptamer and AAs, generating a significant decline in the differential pulse voltammetry (DPV) signals. The proposed aptasensor presented a wide linear range and the detection limit was 3.51 nM. Moreover, the aptasensor, with satisfactory stability, reproducibility and specificity, was successfully employed to detect the multi-residuals of AAs in milk. This work provided a novel strategy for monitoring AAs in milk.

Ultrasensitive electrochemiluminescence aptasensor based on ABEI reduced silver nanoparticles for the detection of profenofos.

An ultrasensitive electrochemiluminescence (ECL) aptasensor for detection of profenofos was constructed by the reducibility and chemiluminescence property of N-(aminobutyl)-N-(ethylisoluminol) (ABEI). ABEI was used to reduce silver nitrate (AgNO3) to silver nanoparticles (AgNPs), which could be adsorbed on the lattice of graphene oxide (GO) to form ABEI-AgNPs-GO complex. This compound could achieve excellent luminescence. The aptamer (Apt) modified (5') by sulfhydryl groups could be immobilized on AgNPs to capture profenofos. When profenofos was present, the ECL signal of the aptasensor would be weakened. To further demonstrate the successful construction of the aptasensor, cyclic voltammetry tests were performed on an electrochemical workstation and an ECL analyzer, respectively. The standard curve and specificity experiment both showed that the sensor had the advantages of low limit of detection (LOD) and good specificity. Under the optimal conditions, the aptasensor had a good linear response for profenofos in the range of 1 × 10-1-1 × 104 ng/mL. It also had a LOD of 6.7 × 10-2 ng/mL and a correlation coefficient (R2) of 0.9991. The aptasensor had been successfully applied to the detection of profenofos in vegetables. The recovery range of the proposed ECL aptasensor was 98 % ~ 107.4 %.

Construction of a dual-model aptasensor based on G-quadruplexes generated via rolling circle amplification for visual/sensitive detection of kanamycin.

A dual-model colorimetric and electrochemical aptasensor was designed using a large number of G-quadruplexes generated by rolling circle amplification (RCA). Specific binding between target and aptamer during RCA yielded large numbers of G-quadruplexes. A colorimetric sensor was fabricated based on the interaction between the G-quadruplex and hemin, which altered the 3,3',5,5'-Tetramethylbenzidine (TMB)-catalyzed color reaction and facilitated the visual and semi-quantitative detection of kanamycin. An electrochemical sensor was constructed based on the strong interaction between the G-quadruplex and the methylene blue electrical signal molecule. Combining nanocomposites multi-walled carbon nanotubes-chitosan/gold nanoparticles (MWCNTs-CS/AuNPs) and RCA realized double-amplified electrochemical signals. Under optimized conditions, a linear relationship was obtained as the logarithm of different concentrations of kanamycin (KAN). The colorimetric aptasensor had a linear range of 1 × 102 nM to 1 × 103 nM with a detection limit of 1.949 nM. The electrochemical aptasensor had wider a linear range from 1 × 10-3 nM to 2.5 × 103 nM and a lower detection limit of 0.333 pM. The sensor combined the advantages of simple colorimetric visualization with the ultra-precision of electrochemical methods. Aptasensor showed good specificity and prevented interference. Furthermore, the prepared dual-model aptasensor facilitated the practical monitoring of KAN in milk.

Novel sandwich-type electrochemiluminescence aptasensor based on luminol functionalized aptamer as signal probe for kanamycin detection.

A novel sandwich electrochemiluminescence (ECL) aptasensor was developed for highly sensitive detection of kanamycin using luminol-functionalized aptamer as a signal probe. The aptasensor used polyethyleneimine (PAMAM), molybdenum disulfide, and multi-walled carbon nanotubes as the substrate, which provided enough binding sites for aptamer1 (the aptamer which modified NH2) coupling. We found that kanamycin could be detected using the aptamer1 containing the same base sequence as aptamer2 (the aptamer which modified SH) on the electrode self-assembly. In addition, PAMAM nanocomposites can be used to effectively improve the ECL intensity by loading a high volume of luminol molecules and silver nanoparticles. In the presence of kanamycin, the sandwiched aptasensor was formed between aptamer1 and the probe of aptamer2 connecting silver nanoparticles, luminol, and PAMAM, resulting in a proportional increase of ECL intensity. Since the significantly enhanced loading of luminol by PAMAM accelerated the electron transfer, the sensitive aptasensor exhibited a wide linear range of detection from 1 × 10-3 to 1 × 103 ng/mL and a low detection limit of 0.21 pg/mL (S/N) for kanamycin. The fabricated aptasensor was successfully applied in quantitative analysis of kanamycin in milk samples.

Non-immobilized GO-SELEX of aptamers for label-free detection of thiamethoxam in vegetables.

Due to the massive use of thiamethoxam (TMX) pesticide and the accumulated potential hazards exposure, the detection of TMX is of great significance to food and ecological safety. In this study, aptamers with affinity for TMX were obtained through graphene oxide assisted systematic evolution of ligands by exponential enrichment (GO-SELEX). After 9 rounds of positive and counter selection, 5 candidate sequences were obtained, among which seq.20 had the highest affinity for TMX, and its dissociation constant (Kd) was 210.47 ± 79.37 nM. Then, the aptamer was further truncated based on structural analysis. The truncated aptamers (seq.20-1, seq.20-2) exhibited higher affinity (Kd = 118.34 ± 13.85 nM, Kd = 123.35 ± 29.80 nM), which seq.20-2 had only 37 bases. Furthermore, circular dichroism spectroscopy showed that TMX induced the conformation of aptamer from B-form structure to hairpin structure, and then formed a stable TMX-ssDNA complex. Finally, the truncated aptamer (seq.20-2) and the original aptamer (seq.20) were used as recognition elements to construct colorimetric aptasensors based on gold nanoparticles for the detection of TMX. It was found that the sensitivity of the former (LOD = 1.67 ± 0.12 nM, S/N = 3) was better than that of the latter (LOD = 3.33 ± 0.23 nM, S/N = 3). Feasibility of truncated aptamer as recognition element in the detection of TMX in vegetable samples was preliminarily verified.

Indirect signal amplification strategy with a universal probe-based lateral flow immunoassay for the rapid quantitative detection of fumonisin B1.

Fumonisin B1 (FB1) is a serious threat to the health of humans and animals. Herein, a lateral flow immunoassay based on universal detection probes (goat anti-mouse IgG@Eu) that could combine with any mouse monoclonal antibody was applied to detect FB1 in corn and feed. Compared with that based on direct monoclonal antibody labeling, this assay maintained bioactivity and saved consumption of monoclonal antibodies with the indirect signal amplification effect. The results indicated that this assay had higher sensitivity with a limit of detection (LOD) of 0.025 and 0.097 ng mL-1 (0.50 and 1.94 ng g-1 based on sample weight) in corn and feed, respectively. The detection range was about 1-50 ng mL-1 (20-1000 ng g-1 based on sample weight). In addition, the evaluation proved that it had good specificity, accuracy, precision, and applicability, and thus was suitable for the rapid and low-cost detection of fumonisin B1.

Recent advances in aptamer-based sensors for aminoglycoside antibiotics detection and their applications.

Aminoglycoside antibiotics (AAs) have been extensively applied in medical field and animal husbandry owing to desirable broad-spectrum antibacterial activity. Excessive AAs residues in the environment can be accumulated in human body through food chain and cause detrimental effect on human health. The establishment of highly specific, simple and sensitive detection methods for monitoring AAs residues is highly in demand. Aptasensor using aptamer as the biological recognition element is the efficient and promising sensing method for detection of AAs. In this review, we have made a summary of specific aptamers sequences against AAs. Subsequently, we provide a systematical and comprehensive overview of modern techniques in aptasensors for detection of AAs according to optical aptasensors as well as electrochemical aptasensors and further summarize their advantages and disadvantages to compare their applications. In addition, we present an overview of practical applications of aptasensors in sample detection of AAs. Moreover, the current challenges and future trends in this field are also included to reveal a promising perspective for developing novel aptasensors for AAs.

Improving lipid productivity by engineering a control-knob gene in the oleaginous microalga Nannochloropsis oceanica.

Nannochloropsis spp. are promising industrial microalgae for scalable oil production and the lipid production can be boosted by nutrient starvation and high irradiance. However, these stimuli halt growth, thereby decreasing overall productivity. In this study, we created transgenic N. oceanica where AtDXS gene encoding 1-deoxy-D-xylulose 5-phosphate synthase (DXS) derived from Arabidopsis thaliana was overexpressed in vivo. Compared with the wild type (WT), engineered Nannochloropsis showed a higher CO2 absorption capacity and produced more biomass, lipids, and carbohydrates with more robust growth in either preferred conditions or various stressed conditions (low light, high light, nitrogen starvation, and trace element depletion). Specifically, relative to the WT, lipid production increased by ~68.6% in nitrogen depletion (~1.08 â€‹g â€‹L-1) and ~110.6% in high light (~1.15 â€‹g â€‹L-1) in the transgenic strains. As for neutral lipid (triacylglycerol, TAG), the engineered strains produced ~93.2% more in nitrogen depletion (~0.77 â€‹g â€‹L-1) and ~148.6% more in high light (~0.80 â€‹g â€‹L-1) than the WT. These values exceed available records in engineered industrial microalgae. Therefore, engineering control-knob genes could modify multiple biological processes simultaneously and enable efficient carbon partitioning to lipid biosynthesis with elevated biomass productivity. It could be further exploited for simultaneous enhancement of growth property and oil productivity in more industrial microalgae.

Voltammetric determination of organophosphorus pesticides using a hairpin aptamer immobilized in a graphene oxide-chitosan composite.

An aptasensor is described for electrochemical determination of organophosphorus pesticides (OPPs), specifically of profenofos, phorate, isocarbophos, and omethoate. The method uses a hairpin aptamer as signalling donor. Its 5' and 3' ends were modified with amino groups and the redox probe ferrocene (Fc), respectively. A nanocomposite consisting of graphene oxide and chitosan (GO-chit) was used to immobilize the aptamer via formation of an amide link. Its good conductivity facilitates monitoring of the electrochemical responses. Upon addition of an OPP, it will be bound by the aptamer. This results in an opening of the hairpin structure. Thus, Fc is shifted away from the surface of the electrode. As a result, the impedance increases and the redox signal of Fc decreases. The electrochemical performance, binding capacity and response of the aptasensor for profenofos, phorate, isocarbophos and omethoate were studied. The limits of detection are as low as 0.01, 0.1, 0.01 and 0.1 nM, respectively. Graphical abstract Schematic representation of an electrochemical aptasensor prepared by immobilizing ferrocene (Fc) labeled hairpin aptamer (HP) on the surface of graphene oxide-chitosan (GO-chit) modified electrode, and its application to the determination of organophosphorus pesticides (OPPs) by voltammetry.