Rapid Selection Research Articles

Study on aptamer based high throughput approach identifies natural ingredients against RGNNV.

Nervous necrosis virus (NNV) is one of the most destructive pathogens in marine fish aquaculture and is capable of infecting more than 50 fish species worldwide, which resulted in great economic losses. Effective drugs for managing NNV infection are urgently required. Medicinal plants have been known for thousands of years and benefit of medicinal plants against pathogens in aquaculture have emerged. Nowadays, the most commonly used method for detecting virus infection and assessing antiviral drugs efficacy is reverse transcription-quantitative real-time PCR. However, the application is limited on account of high reagent costs, complex time-consuming operations and long detection time. Aptamers have been widely applied in application of pathogens or diseases diagnosis and treatments because of high specificity, strong affinity, good stability, easy synthesized and low costs. This study aimed to establish an aptamer (GBN34)-based high-throughput screening (GBN34-AHTS) model for efficient selection and evaluation of natural ingredients against NNV infection. GBN34-AHTS is an expeditious rapid method for selecting natural ingredients against NNV, which is characterized with high-speed, dram, sensitive and accurate. AHTS strategy could reduce work intensity and experimental costs and shorten the whole screening cycle of effective ingredients. AHTS should be suitable for rapid selection of effective ingredients against other viruses, which is important for improving the prevention and controlling of aquatic diseases.

A highly selective probe for ratiometric imaging peroxynitrite in living cells and in vivo

In this study, we constructed and displayed a ratiometric fluorescent probe JQ-2 for detecting ONOO−. The probe JQ-2 showed a ratiometric signal for visualizing ONOO− with a rapid response and high selectivity over a panel of biological analytes. Moreover, the JQ-2 has near-infrared emission (657 nm), which provides an excellent basis for the practical application in biological systems. The probe JQ-2 possessed low cytotoxicity and excellent cell membrane permeability, which can specifically visualize the exogenous and endogenous ONOO− in vitro and vivo by emission in two channels. Meanwhile, JQ-2 can be used for diagnosing drug-induced liver injury by visualizing and monitoring the fluctuations of endogenous ONOO−. Therefore, JQ-2 provided a potential tool for precisely detecting the fluctuation of ONOO− in biological systems to understand physiological and pathological process.

The construction of a ratiometric dual-emitting amorphous europium-organic frameworks for sensitive detection of water in white spirit

It is very crucial to develop a simple and fast-responsive sensor for the detection of water content in organic solvents. Herein, three different europium-organic frameworks (Eu-MOFs): amorphous Eu-2-aminoterephthalic acid (amEu-NH2BDC), crystalline Eu-2-aminoterephthalic acid (cryEu-NH2BDC) and amorphous Eu-1,4-benzenedicarboxylic acid (amEu-BDC) are synthesized, respectively. amEu-NH2BDC is employed as fluorescence probe for the determination of water in ethanol. Results indicate that amEu-NH2BDC exhibits good performance due to its dual-wavelength fluorescence (FL) with the ligand (NH2BDC)-centered blue luminescence at 430 nm and Eu-centered red luminescence at 615 nm. With the increase of water contents in ethanol, the emission intensity at 430 nm increases dramatically, while that at 615 nm decreases under a single excitation at 270 nm. By measuring the ratio of emission intensity at 430 nm and 615 nm (I430/I615), a wide concentration range from 1% to 99.5% water in ethanol is obtained with a lower detection limit value of 0.024% (S/N = 3). Besides, in the presence of water in ethanol, the fluorescence color of amEu-NH2BDC changes from red to blue under 365 nm UV light. Thus, the visual device for the detection of water is realized by using amEu-NH2BDC. Combined with the fluorescence lifetime and density functional theory, it is shown that red luminescence weakens due to dynamic quenching and non-radiative vibration energy transfer, and blue luminescence is enhanced due to the hydrogen bonding between water and carboxyl groups in the ligand, which enhances the intramolecular charge transfer process. The proposed method has characteristics of rapid response, excellent stability, high sensitivity and selectivity. Importantly, it can be successfully applied to monitor the content of water in white spirit samples.

A highly selective and sensitive fluorescence probe based on Rhodol for imaging of endogenous peroxynitrite in living cells

Peroxynitrite (ONOO−), as one of the important reactive oxygen/nitrogen species, is associated with many physiological and pathological activities. Therefore, effective detection methods with high sensitivity and rapid response time are essential to reveal its functions. A novel Rhodol-based fluorescent probe RN is constructed and synthesized by introducing of 1,1-dimethylhydrazone as a recognition group. Due to the oxidative breakage of the CN bond by ONOO−, the probe has significant fluorescence enhancement change with excellent detection properties such as rapid response time, high sensitivity and selectivity. Moreover, probe RN show low cytotoxicity and is successfully capable of imaging endogenous and endogenous ONOO− in living cells. These properties indicate that RN has great application potential for imaging of ONOO− in biological systems.

A Turn-On Fluorescent Chemosensor for Cyanide Ion Detection in Real Water Samples.

We have designed and synthesized a novel simple colorimetric fluorescent probe with aggregation-induced emission (AIE) properties. Probe 5-(4-(diphenylamine)phenyl) thiophen-2-formaldehyde W exhibited a turn-on fluorescent response to cyanide ion (CN−), which induces distinct visual color changes. Probe W exhibited a highly selective and sensitive ratiometric fluorescence response for the detection of CN− over a wide pH range (4–11) and in the presence of common interferents. The linear detection of CN− over the concentration range of 4.00–38.00 µM (R 2 = 0.9916, RSD = 0.02) was monitored by UV-Vis absorption spectrometry (UV-Vis) with the limit of detection determined to be 0.48 µM. The linear detection of CN− over the concentration range of 8.00–38.00 µM was examined by fluorescence spectrophotometry (R 2 = 0.99086, RSD = 0.031), and the detection limit was found to be 68.00 nM. The sensing mechanisms were confirmed by 1H NMR spectroscopic titrations, X-ray crystallographic analysis, and HRMS. Importantly, probe W was found to show rapid response, high selectivity, and sensitivity for cyanide anions in real water samples, over the range of 100.17∼100.86% in artificial lake water and 100.54∼101.64% in running water by UV-Vis absorption spectrometry, and over the range of 99.42∼100.71% in artificial lake water and 100.59∼101.17% in running water by fluorescence spectrophotometry. Importantly, this work provides a simple and effective approach which uses an economically cheap and uncomplicated synthetic route for the selective, sensitive, and quantitative detection of CN− ions in systems relevant to the environment and health.

Efficient Method for Generating Point Mutations in the Vaccinia Virus Genome Using CRISPR/Cas9.

The vaccinia virus (VACV) was previously used as a vaccine for smallpox eradication. Nowadays, recombinant VACVs are developed as vaccine platforms for infectious disease prevention and cancer treatment. The conventional method for genome editing of the VACV is based on homologous recombination, which is poorly efficient. Recently, the use of CRISPR/Cas9 technology was shown to greatly improve the speed and efficiency of the production of recombinant VACV expressing a heterologous gene. However, the ability to rapidly recover viruses bearing single nucleotide substitutions is still challenging. Notwithstanding, ongoing studies on the VACV and its interaction with the host cell could benefit from viral gene targeted mutagenesis. Here, we present a modified version of the CRISPR/Cas9 system for the rapid selection of mutant VACV carrying point mutations. For this purpose, we introduced a silent mutation into the donor gene (which will replace the wildtype gene) that serves a double function: it is located in the PAM (NGG) sequence, which is essential for Cas9 cleavage, and it alters a restriction site. This silent mutation, once introduced into the VACV genome, allows for rapid selection and screening of mutant viruses carrying a mutation of interest in the targeted gene. As a proof of concept, we produced several recombinant VACVs, with mutations in the E9L gene, upon which, phenotypic analysis was performed.

A study on the optimal selection of similar materials for the physical simulation experiment based on rock mineral components

The physical model test can effectively solve the problems of great difficulty and the high cost of the field test. Similarity theory is an essential basis for model test design. Accurate and rapid selection, proportioning, and production of similar materials are of great significance in accurately simulating the project site's rock stratum geological situation. However, the existing preparation methods of similar materials have great blindness and a low success rate. Considering the critical influence of mineral composition on the physical and mechanical properties of rock, aiming at preparing rock sample materials in TBM tunneling simulation under high ground stress, this paper takes Sichuan white sandstone as the specific research object. We use mineral composition to guide similar materials' mix design and preparation. It is found that similar materials with the required geometric similarity ratio can be obtained by adjusting the water-cement ratio and cement content under the condition that the mineral composition is basically the same. The similar material sample of Sichuan white sandstone with a given geometric similarity ratio (a(a＞1.5)) can be designed with cement content (C) as C = 150a-1 and water cement ratio (Wrt) as Wrt = 0.072a + 0.108. We believe this method can also provide a reference for preparing similar materials for other rocks.

A novel protein-based supramolecular recognition approach for ratiometric fluorescence detection of fipronil

Fipronil, a wildly used pesticide, exhibits high toxicity to pollinators, aquatic organisms, farmland birds, as well as mammals that have GABA receptors, including humans. Thus, developing an effective method for fipronil determination is of great importance. Although the fluorescence assays have the features of easy-to-use, rapid response, high sensitivity and visualized readout, such assay for fipronil was rather difficult to design because fipronil was both chemically and photophysically stable. Herein, we utilized a chalcone dye bound into albumin inner-cavity to construct a novel supramolecular recognition system, for the first time, successfully realizing ratiometric fluorescence detection of fipronil. The sensing mechanism has been carefully verified by the molecular docking, fluorescent titration, DLS tests and fluorescence lifetime. This reported probe could detect fipronil with a rapid response (< 3 min), high sensitivity (LOD ~ 0.01 ppm) and selectivity, good anti-interference property, and significantly, ratiometric color change from green to yellow, which could be discriminated by naked eyes. Moreover, we demonstrated the application of this method in the ratiometric tracking of fipronil in plants (Arabidopsis thaliana). We anticipate this albumin-based supramolecular recognition system would inspire more probe design for environmental monitoring.

Novel magnetic covalent organic framework loaded ligand for rapid removal and selective detection of mercury(II) from water

The present work aimed to prepare a novel magnetic covalent organic framework loaded ligand (MCM), which is going to be utilized to obtain efficient adsorption, rapid removal, and detection with high selectivity and sensitivity for mercury(II) ions. The structure, morphology, adsorption behavior, and detection properties of MCM were characterized with FTIR, TG, XRD, BET, SEM, TEM, UV-VIS, and XPS. To optimize the effects of process parameters, the influence of pH, contact time, and initial concentration on the adsorption behavior, color optimization, and detection limit of the mercury(II) ions was investigated. Under the optimal condition, MCM showed the efficient adsorption ability, rapid removal rate, high selectivity, and detection with excellent sensitivity for the mercury(II) ions. The color of MCM changed from light grey to yellow by observing with naked-eye in the presence of the mercury (II) ions, and the corresponding detection limit was 0.01μg/L. MCM also has a high removal rate of mercury (II) ions in industrial wastewater, which can meet the industrial discharge standard (residual concentration of mercury(II) ions in the solution is 0.05 mg/L). What's more, MCM could be used in a portable column unit to separate mercury(II) ions from actual water samples, rapidly reducing the mercury(II) concentration from 10.0 mg/L to 10.0 μg/L within 15 s. Kinetic studies showed that the MCM adsorption behavior for mercury(II) ions was in good agreement with the pseudo-second-order model, and the MCM adsorption equilibrium data for mercury(II) ions belonged to the Langmuir model. Furthermore, the adsorption amount of MCM for mercury(II) ions after five adsorptions was 96.89% of the adsorption amount of MCM for the first adsorption of mercury(II) ions, which showed that MCM has good regeneration ability. Consequently, MCM provided an excellent alternative to mercury(II) ions adsorption or enrichment materials, and this method showed great potential in the routine monitoring of trace mercury(II) ions in liquid substrates.

PNGSeqR: An R Package for Rapid Candidate Gene Selection through Pooled Next-Generation Sequencing

Although bulked segregant analysis (BSA) has been used extensively in genetic mapping, user-friendly tools which can integrate current algorithms for researchers with no background in bioinformatics are scarce. To address this issue, we developed an R package, PNGSeqR, which takes single-nucleotide polymorphism (SNP) markers from next-generation sequencing (NGS) data in variant call format (VCF) as the input file, provides four BSA algorithms to indicate the magnitude of genome-wide signals, and rapidly defines the candidate region through the permutation test and fractile quantile. Users can choose the analysis methods according to their data and experimental design. In addition, it also supports differential expression gene analysis (DEG) and gene ontology analysis (GO) to prioritize the target gene. Once the analysis is completed, the plots can conveniently be exported.

Genetic and structural basis of the human anti-α-galactosyl antibody response

Humans lack the capacity to produce the Galα1-3Galβ1-4GlcNAc (α-gal) glycan, and produce anti-α-gal antibodies upon exposure to the carbohydrate on a diverse set of immunogens, including commensal gut bacteria, malaria parasites, cetuximab, and tick proteins. Here we use X-ray crystallographic analysis of antibodies from α-gal knockout mice and humans in complex with the glycan to reveal a common binding motif, centered on a germline-encoded tryptophan residue at Kabat position 33 (W33) of the complementarity-determining region of the variable heavy chain (CDRH1). Immunoglobulin sequencing of anti-α-gal B cells in healthy humans and tick-induced mammalian meat anaphylaxis patients revealed preferential use of heavy chain germline IGHV3-7, encoding W33, among an otherwise highly polyclonal antibody response. Antigen binding was critically dependent on the presence of the germline-encoded W33 residue for all of the analyzed antibodies; moreover, introduction of the W33 motif into naive IGHV3-23 antibody phage libraries enabled the rapid selection of α-gal binders. Our results outline structural and genetic factors that shape the human anti-α-galactosyl antibody response, and provide a framework for future therapeutics development.

Size-Controlling Cherry Rootstock Selection Based on Root Anatomical Characteristics

Over the past few decades, numerous studies investigated the vigor and productivity of fruit species depending on the rootstock on which they were grafted, but the exact size-controlling mechanism itself has not been fully elucidated, nor were the rapid rootstock selection methods defined. Thus, this study aimed to assess the root anatomical characteristics and their influence on the overall ‘Summit’ cherry tree vigor to confirm the size-controlling effect and establish an effective protocol for rapid rootstock selection. Plant material included three cherry species (Prunus cerasus, Prunus fruticosa, and Prunus mahaleb) and interspecific hybrid ‘Gisela 5′ (P. cerasus × Prunus canescens) as a control. The detailed anatomical analysis included root samples with the differentiated secondary structure taken from the sampling depth of 10–15 cm. Roots with percentages of vessels ≈40%, ≈50%, and ≈10% belonging to size-classes ˂700 µm2, 700–2000 μm2, and ˃2000 µm2 (respectively) are presumed to provide optimal amounts of water solution to the scion, without compromising plant vitality, drought tolerance, and size-controlling effect. Statistically significant correlations were determined between anatomical properties (the percentage of vessels, especially ˃2000 µm2, xylem porosity, and hydraulic conductivity, both per mm2 and total root) and vegetative growth in the juvenile vegetative phase, indicating direct vessel size influence on plant vigor and its employment in size-controlling cherry rootstock selection.

The early attentional pancake: Minimal selection in depth for rapid attentional cueing.

There have been conflicting findings on the degree to which rapidly deployed visual attention is selective for depth, and this issue has important implications for attention models. Previous findings have attempted to find depth-based cueing effects on such attention using reaction time (RT) measures for stimuli presented in stereo goggles with a display screen. Results stemming from such approaches have been mixed, depending on whether target/distractor discrimination was required. To help clarify the existence of such depth effects, we have developed a paradigm that measures accuracy rather than RT in an immersive virtual-reality environment, providing a more appropriate context of depth. Three modified Posner Cueing paradigms were run to test for depth-specific rapid attentional selectivity. Participants fixated a cross while attempting to identify a rapidly masked black letter preceded by a red cue that could be valid in depth, side, or both. In Experiment 1a, a potent cueing effect was found for lateral cueing validity, but a weak effect was found for depth despite an extreme difference in virtual depth (1 vs. 300 m). In Experiment 1b, a near-replication of 1a, the lateral effect replicated while the depth effect did not. Finally, in Experiment 2, to increase the depth cue's effectiveness, the letter matched the cue's color, and the presentation duration was increased; however, again only a minimal depth-based cueing effect - no greater than that of Experiment 1a - was observed. Thus, we conclude that rapidly deployed attention is driven largely by spatiotopic rather than depth-based information.

Biomass yield improvement in switchgrass through genomic prediction of flowering time

AbstractThe seasonal timing of the transition from vegetative to reproductive growth has a major impact on biomass accumulation in switchgrass. Late‐flowering switchgrass cultivars produce greater biomass, a critical trait for sustainable bioenergy production. Genomic prediction (GP) may allow rapid selection of late‐flowering individuals with reduced time and expense for field evaluations. To evaluate GP, two flowering time traits (heading date and anthesis date) were collected on 1532 genotypes from four breeding populations: Midwest, Gulf, Atlantic, and Hybrid. These were sequenced using genotype‐by‐sequencing (530,792 single‐nucleotide polymorphisms). Predictive ability of single‐trait and multi‐trait models were evaluated by cross‐validation, by prediction of a progeny trial (n = 122), and through prediction of yield performance in a parallel experiment (n = 52). Predictive ability was not improved by sharing information among breeding groups. Overall, multi‐trait models provided an advantage during cross‐validation, but a smaller advantage during progeny prediction. Within populations, GP resulted in lower per‐cycle progress than previously reported field evaluations (3.1 vs. 5.0 day−1 cycle−1). However, GP cycles are potentially much faster than field evaluations. When directly predicting biomass yield, the Hybrid training population had a predictive ability of 0.54–0.63. This reinforces the strong linkage between biomass yields in swards and flowering time. These results highlight the value of GP for rapid yield improvement in switchgrass, particularly in a breeding program designed to share information between biomass yield trials and low‐cost flowering time evaluations.

Infectious Bronchitis Coronavirus: Genome Evolution in Vaccinated and Non-Vaccinated SPF Chickens.

Infectious Bronchitis virus (IBV) continues to cause significant economic losses for the chicken industry despite the use of many live IBV vaccines around the world. Several authors have suggested that vaccine-induced partial protection may contribute to the emergence of new IBV strains. In order to study this hypothesis, three passages of a challenge IBV were made in SPF chickens sham inoculated or vaccinated at day of age using a live vaccine heterologous to the challenge virus. All birds that were challenged with vaccine heterologous virus were positive for viral RNA. NGS analysis of viral RNA in the unvaccinated group showed a rapid selection of seven genetic variants, finally modifying the consensus genome of the viral population. Among them, five were non-synonymous, modifying one position in NSP 8, one in NSP 13, and three in the Spike protein. In the vaccinated group, one genetic variant was selected over the three passages. This synonymous modification was absent from the unvaccinated group. Under these conditions, the genome population of an IBV challenge virus evolved rapidly in both heterologous vaccinated and non-vaccinated birds, while the genetic changes that were selected and the locations of these were very different between the two groups.

Abstract 4104: Low-cost, simple and rapid assay for single-molecule detection of gene fusions from RNA with ASPYRE technology

Abstract Introduction: The FDA has approved 11 therapies for non-small cell lung cancer (NSCLC) patients diagnosed with gene fusions. Importantly, these life-saving targeted therapies remain underutilized due to a lack of cost-effective, comprehensive, and simple-to-use technologies for gene fusion detection. Our previous work using ASPYRE technology (allele-specific pyrophosphorolysis reaction; Silva et al, 2020) demonstrated detection of somatic variants at single-molecule level from DNA extracted from tumor tissue. However, DNA-based fusion detection requires tiling across large introns, which limits sensitivity particularly if breakpoints are located in repetitive regions of the genome. Here, we show that ASPYRE technology can also be utilized for highly sensitive detection of RNA fusions, scaling to over 30 fusion targets covering the main 3’ fusion partners (ALK, ROS1, RET, NTRK1, NTRK3) and from a single reverse transcription PCR (RT-PCR) amplification reaction. Methods: The ASPYRE assay consists of four sequential enzymatic stages: RT-PCR; enzymatic clean-up; combined exonuclease digestion, hybridization, pyrophosphorolysis and ligation; rolling circle isothermal amplification and detection. All targets are detected across two reaction wells using four channels on a standard real-time PCR instrument. A panel of synthetic RNA oligonucleotides comprising gene fusion boundaries was spiked into a background pool of lung-RNA, quantified using digital PCR (dPCR) and compared against a commercial reference standard to validate its suitability for assessing assay performance. RNA was extracted from formalin-fixed paraffin-embedded (FFPE) lung tissue from healthy donors to determine assay specificity, and from FFPE lung tissue from NSCLC patients with known fusions to exemplify end-to-end sample analysis. Results: We demonstrate equivalence of our in-house reference samples to commercial reference standards. Using serial dilutions of input RNA standards, we show that detection by ASPYRE is consistent with detection of single molecules as input for common ROS1, ALK, RET and NTRK3 fusions. In addition, ASPYRE was able to detect previously confirmed ROS1 and ALK fusions and MET exon skipping variants in FFPE samples. ASPYRE had high specificity, correctly confirming the absence of RNA fusions and exon skipping variants in healthy control FFPE samples. Conclusions: ASPYRE approached single-molecule sensitivity for over 30 RNA fusion targets in a single reaction. The assay required only 1 ng RNA input, with a turn-around time of &amp;lt;4 hours from extracted RNA to result, opening the door to routine detection of RNA fusions from limited specimens. The simplicity and ease of use of ASPYRE, combined with its rapid turnaround time and analytical performance characteristics, provides a new tool for rapid and comprehensive treatment selection in NSCLC. Citation Format: Eleanor Gray, Justyna Mordaka, Efthimia Christoforou, Kristine von Bargen, Nicola Potts, Rebecca Palmer, Barnaby Balmforth. Low-cost, simple and rapid assay for single-molecule detection of gene fusions from RNA with ASPYRE technology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 4104.

Combined Technology Selection Model for Digital Transformation in Manufacturing: A Case Study From the Automotive Supplier Industry

Among generic technology management activities, rapid technology identification and selection stand as the significant determinants of technology adoption success in the digital transformation era. Especially for manufacturing SMEs in developing countries, rapid digital technologies are critical since they struggle to protect their competitiveness in global value chains threatened by digitalization. Previous studies introduce various multi-criteria decision-making model-based approaches to identify and select appropriate manufacturing technologies. However, these approaches were relatively rigid and required an advanced understanding of the technology for criteria and alternative settings and evaluation. Decision-makers need more flexible and scalable contextual frameworks for technology selection in digitalization. Since digital technologies offer both benefits and challenges, the decision-making models should reflect this dialectic nature of Industry 4.0 adoption and contextually optimize their decisions by combining multiple quantitative methods for technology identification and selection. Besides, case studies on digital technology selection are rare in manufacturing SMEs from developing country context in the literature. In this context, this study proposes a technology selection framework that utilizes the three dimensions (industry 4.0 technologies, benefits, and challenges) and combines AHP with a QFD-inspired intervention matrix and an optimization model by Mixed Integer Programming (MIP). The proposed model is validated with a case study from the automotive supplier industry in Turkey with the data provided from interviews and a Delphi survey with 11 experts from the digitalization value chain of the selected industry. Case study results revealed that the highest benefits of industry 4.0 lie in process/quality efficiency improvement and reduced inventory. At the same time, data analytics and sensor technologies occurred as the most critical tools. Significant challenges of digital technology adoption are insufficient expert know-how and budget constraints.

Genetic analysis of leaf trichome density using mixed major gene plus polygene inheritance model in Brassica rapa ssp. chinensis

ABSTRACT Leaf trichome is an important trait for stress-resistance breeding in Brassica crops, but its scientific evaluation and identification are rarely reported. To investigate the inheritance of leaf trichome in non-heading Chinese cabbage, two inbred lines, ‘14XS23W’ (glabrous) and ‘14XS23Y’ (hirsute), were used to generate F1 and F2 populations for joint analysis using a mixed major genes plus polygene inheritance model. The density of adaxial leaf trichomes was controlled by two pairs of additive-dominance-epistasis major genes plus additive-dominance polygenes. The broad-sense heritabilities of the major genes were 93.8% and 85.51%, respectively, for greenhouse and open-field conditions. However, the density of the abaxial leaf trichome was controlled by two pairs of additive-dominance-epistasis major genes plus additive dominance-epistasis polygenes. The broad-sense heritabilities of the major genes were 91.45% and 72.20%, while the polygenes were 0% and 14.5%, respectively, for greenhouse and open field environments. These results indicated that trichome density in non-heading Chinese cabbage was primarily controlled by two pairs of major genes with high heritability, implying that this trait is suitable for rapid selection in early generations.

Aptamer-Based High-Throughput Screening Model for Efficient Selection and Evaluation of Natural Ingredients against SGIV Infection

Singapore grouper iridovirus (SGIV) causes high economic losses in mariculture. Effective drugs for managing SGIV infection are urgently required. Medicinal plant resources are rich in China. Medicinal plants have a long history and significant curative effects in the treatment of many diseases. Reverse-transcription quantitative real-time PCR is the most commonly used method for detecting virus infection and assessing antiviral efficacy with high accuracy. However, their applications are limited due to high reagent costs and complex time-consuming operations. Aptamers have been applied in some biosensors to achieve the accurate detection of pathogens or diseases through signal amplification. This study aimed to establish an aptamer-based high-throughput screening (AHTS) model for the efficient selection and evaluation of medicinal plants components against SGIV infection. Q2-AHTS is an expeditious, rapid method for selecting medicinal plant drugs against SGIV, which was characterized as being dram, high-speed, sensitive, and accurate. AHTS strategy reduced work intensity and experimental costs and shortened the whole screening cycle for effective ingredients. AHTS should be suitable for the rapid selection of effective components against other viruses, thus further promoting the development of high-throughput screening technology.

Two novel enzyme-free colorimetric sensors for the detection of glyphosate in real samples

The intensive use and accumulation of glyphosate cause many environmental and ecological problems. There is an urgent need for an effective detection method for glyphosate. Many studies in the literature determine glyphosate by an enzyme-based method, requiring appropriate temperature, pH, substrate and ionic strength. In this study, we presented a simple enzyme-free reaction system. Two new simple Schiff base derivatives, 4-(((3-chloropyridin-4-yl)imino)methyl)-N,N-dimethylaniline (BNP-Cl) and 4-((3-((3-chloropyridin-4-yl)imino)propenyl)-N,N-dimethylaniline (CNP-Cl), were prepared by combining a pyridine moiety with a cinnamaldehyde or a benzaldehyde moiety. The pyridine moiety contains a chlorine atom, and this chlorine atom is replaced by the nitrogen or oxygen of glyphosate (Glyp) in ACN/Tris buffer (pH=8.14, 10 mM, v/v 2:1). This binding caused color changes ranging from colorless to yellow for BNP-Cl and yellow to orange for CNP-Cl. The sensors BNP-Cl and CNP-Cl have a rapid and excellent selectivity for glyphosate with low detection limits (1.78 µM for BNP-Cl and 1.60 µM for CNP-Cl). The sensors were also successfully applied to detect Glyp in real samples such as soil, tap water, and potatoes.