Biological Amplification Research Articles

An NMR immunosensor based on streptavidin-biotin signal amplification for rapid detection of Salmonella in milk

Sensitive and accurate detection of harmful pathogenic bacteria in food is of great significance to public health. In this paper, a nuclear magnetic resonance (NMR) sensor based on biological signal amplification was constructed. The carboxymethyl dextran (CMD) was used as both the probe skeleton and the chelating agent to realize the fast chelating adsorption of the signal source gadolinium ion. Finally, the magnetic complex and biotinylated antibody formed an immune probe through the biological bridge to achieve rapid capture of Salmonella. In addition, non-target bacteria such as Proteus vulgaris and Listeria monocytogenes were used to test the specificity of the constructed sensor. The sensor can detect Salmonella within 1.5 h under optimal conditions, the detection limit in milk is 7.4 × 103 CFU mL−1, and the RSD (Relative standard deviation) of reproducibility experiment is 2.35%, which has good reproducibility.

Toxicological assessment of cadmium exposure through Hyphantria cunea larvae on the predation fitness of Arma chinensis

Heavy metals are defined as an abiotic factor that affects the efficiency of biological pest control. This study constructed a cadmium (Cd)-polluted artificial diets–Hyphantria cunea–Arma chinensis food chain to analyze the effects of Cd exposure on the ability of A. chinensis to control H. cunea. The results revealed that Cd was transferred through the artificial diet to H. cunea larvae and A. chinensis nymphs via a biological amplification effect. After feeding on Cd-accumulated H. cunea larvae, the body weight of A. chinensis nymphs reduced, mortality increased, developmental duration prolonged, and the expression of growth regulatory genes (EX, cycE, and MER) decreased. Cd activated the antioxidant defense system of the nymphs, accompanied by a significant enhancement in the contents of H2O2 and MDA, marked damage to the midgut sub-microstructure, and a remarkable induction in the expression of genes crucial for the mitochondrial pathway/ER stress–apoptosis pathway. Cd significantly diminished the contents of total amino acids, glucose, free fatty acids, and expression of the genes (HK2, PFK, IDH1, and IDH2) essential for the TCA cycle and glycolysis in the nymphs. The preference of the A. chinensis nymphs to Cd-treated H. cunea larvae was evidently reduced. Cd diminished the search-ability, food intake, instantaneous attack rate, and maximum theoretical daily food intake but prolonged the feeding time of the nymphs. Taken together, Cd exposure reduces the ability of A. chinensis nymphs to control H. cunea and provides a new challenge for the efficiency of insect pest control using natural enemies. These findings have important reference value for optimizing pest control strategies in heavy metal polluted areas.

Capillary force-driven reverse-Tesla valve structure for microfluidic bioassays.

Biological assays involve the lysis of biological particles, enzyme reactions, and gene amplification, and require a certain amount of time for completion. Microfluidic chips are regarded as powerful devices for biological assays and in vitro diagnostics; however, they cannot achieve a high mixing efficiency, particularly in some time-consuming biological reactions. Herein, we introduce a microfluidic reverse-Tesla (reTesla) valve structure in which the fluid is affected by vortices and branch flow convergence, resulting in flow retardation and a high degree of mixing. The reTesla is passively operated by a microfluidic capillary force without any pumping facility. Compared with our previously developed micromixers, this innovative pumpless microfluidic chip exhibited high performance, with a mixing efficiency of more than 93%. The versatility of our reTesla chip will play a pivotal role in the study of various biological and chemical reactions.

Glutathione conjugation and protein modification resulting from metabolic activation of pesticide metalaxyl in vitro and in vivo

Metalaxyl (MTL), a germicidal agent, is widely used in agriculture. Due to the biological amplification effect, MTL entering the ecological environment would result in a threat to human health through the food chain. MTL is reportedly accumulated in liver. The objectives of the study included investigating the metabolic activation of MTL in liver and defining the mechanisms participating in the hepatotoxicity of MTL. The corresponding glutathione (GSH), N-acetylcysteine (NAC) conjugate, and cysteine conjugates were observed in liver microsomes, prepared from liver tissues of mice, containing MTL and GSH, NAC or cysteine. These conjugates were also detected in urine and bile of rats receiving MTL. Apparently, MTL was biotransformed to a quinone imine intermediate dose-dependently attacking the thiols and cysteine residues of protein. The bioactivation of MTL required cytochrome P450 enzymes, and CYP3A dominated the bio-activation of MTL.

Smart biocathodic double signal amplification design empowered self-powered bioplatform ultrasensitive detection of tumor biomarker

MicroRNA-21 (miRNA-21) is widely expressed in various organs and tissues of mammals, including the heart, lungs, brain, pancreas, small intestine, colon, breast, and kidneys, showing high sensitivity and specificity in the diagnosis of multiple types of tumors. It can serve as a new clinical early diagnosis and prognostic biomarker for tumors. This study proposes a self-powered bioplatform, which employs a biological cathode double signal amplification design to construct a high-performance enzyme biofuel cell for the ultra-sensitive detection of tumor biomarker miRNA-21. The biocathodic double signal mechanism involves the catalyzed hairpin self-assembly introduction signal I bilirubin oxidase and a tetrahedral DNA nanostructure that produces a double-helical DNA chain to adsorb signal II electron acceptor [Ru(NH3)6]3+. This approach not only improves the stability of the sensing system, but also synergistically enhances the open circuit voltage. The developed assay exhibits a wide linear range from 0.1 fM to 1000 pM, with a low detection limit of 0.04 fM (S/N = 3). This study provides a strategy for designing biosensors with improved sensitivity and potential application in medical diagnosis field.

Cadmium (Cd) exposure through Hyphantria cunea pupae reduces the parasitic fitness of Chouioia cunea: A potential risk to its biocontrol efficiency

Heavy metal contamination has been regarded as an environmental variable that affects the efficiency of pest biological control, but the parasitic fitness of parasitoids under heavy metal stress is poorly understood. Herein, the effect of Cd exposure through the host pupa of Hyphantria cunea on the parasitic fitness of Chouioia cunea was investigated, and the mechanism by which Cd exposure affects the interaction between H. cunea and C. cunea from the perspective of innate immunity in host insect and the oxidative status in the parasitoid offspring was explored. Our results indicated that Cd can be transferred from the H. cunea pupae to the parasitoid offspring, and the transfer coefficient reflected biological amplification. There were no significant differences in the rates of parasitism success and offspring emergence between the untreated and Cd-treated groups. However, after parasitizing Cd-accumulated pupae, the parasitic fitness of offspring wasps (e.g., the number, individual size and life span) decreased significantly. Under Cd exposure, the cellular and humoral immunity of H. cunea pupae decreased significantly. Compared with the untreated group, the H2O2 content of parasitoid offspring in the Cd-treated group was significantly increased. Cd exposure significantly inhibited superoxide dismutase activity in parasitoid offspring, but the contents of ascorbic acid and glutathione were significantly increased by Cd stress. Taken together, these results indicate that Cd exposure reduces the cyclic utilization efficiency of C. cunea on H. cunea pupae. The oxidative status of parasitoid offspring triggered by Cd exposure could be responsible for the reduced parasitic fitness of C. cunea on Cd-accumulated H. cunea pupae.

Biological redox cycling amplification in a self-powered photoelectrochemical sensor based on TiO2/CdIn2S4/ g-C3N4-WO3 photoanode for sensitive detection of Hg2+

A photoelectrochemical (PEC) sensor is proposed with a TiO2/CdIn2S4 co-sensitive structure and a g-C3N4-WO3 heterojunction as the photoanode to form a self-powered system. The photogenerated hole-induced biological redox cycle of TiO2/CdIn2S4/g-C3N4-WO3 composites is used as a signal amplification strategy for Hg2+ detection. In the test solution, ascorbic acid is first oxidized by the photogenerated hole of the TiO2/CdIn2S4/g-C3N4-WO3 photoanode, which triggers the ascorbic acid－glutathione cycle to achieve signal amplification and increase the photocurrent. However, in the presence of Hg2+, glutathione forms a complex with Hg2+, which destroys the biological cycle and leads to a decreased of photocurrent, thus achieving detection of Hg2+. Under optimal conditions, the proposed PEC sensor has a wider range (from 0.1 pM to 100 nM), and lower limit of Hg2+ detection (0.44 fM) than most other Hg2+ detection methods. In addition, the developed PEC sensor can be used to detect of real samples.

Impact of Microplastics and Nanoplastics on Livestock Health: An Emerging Risk for Reproductive Efficiency.

Pollution due to microplastics and nanoplastics is one of the major environmental issues of the last decade and represents a growing threat to human and animal health. In aquatic species, there is a large amount of information regarding the perturbation of marine organisms; instead, there are only a few studies focusing on the pathophysiological consequences of an acute and chronic exposure to micro- and nanoplastics in mammalian systems, especially on the reproductive system. There are several studies that have described the damage caused by plastic particles, including oxidative stress, apoptosis, inflammatory response, dysregulation of the endocrine system and accumulation in various organs. In addition to this, microplastics have recently been found to influence the evolution of microbial communities and increase the gene exchange, including antibiotic and metal resistance genes. Special attention must be paid to farm animals, because they produce food such as milk, eggs and meat, with the consequent risk of biological amplification along the food chain. The results of several studies indicate that there is an accumulation of microplastics and nanoplastics in human and animal tissues, with several negative effects, but all the effects in the body have not been ascertained, especially considering the long-term consequences. This review provides an overview of the possible adverse effects of the exposure of livestock to micro- and nanoplastics and assesses the potential risks for the disruption of reproductive physiological functions.

Effect of weak alternating magnetic fields on neutrophilic granulocytes. An analytical review

The article discusses the main achievements in recent years in studying the biological effects of weak and superweak low-frequency magnetic fields, either variable or combined with constant ones. Considered are neutrophil granulocytes activated by chemical stimulants or intact when the magnetic fields affect isolated cells, blood, and whole organisms. The methods include recording changes in ROS concentration levels (the most noticeable effect of exposure to a weak magnetic field), priming index, calcium homeostasis, proliferative activity, immune status, as well as the influence of various chemical agents on these indicators. The leading methods in this field are fluorescence spectrometry and chemiluminescence analysis. The experimental results indicate the biological effectiveness of this physical factor, the specific effect of which depends on the type of biosystem, its functional status, the environment, and the parameters of the fields themselves. The data obtained can have applied significance in magnetotherapy, immune response optimization in various diseases, acceleration of tissue regeneration and repair, and increasing the body's resistance to infections. They also can have academic significance since they help to identify the primary field acceptors and magnetic targets and their localization in the cell, study relationships with signal cascades, build models of biological signal amplification pathways, and find biologically significant frequencies and field amplitudes.

Chemical Cyclic Amplification: Hydroxylamine Boosts the Fenton Reaction for Versatile and Scalable Biosensing

Nucleic acid detection is undoubtedly one of the most important research fields to meet the medical needs of genetic disease diagnosis, cancer treatment, and infectious disease prevention. However, the practical detection methods based on biological amplification are complex and time-consuming and require highly trained operators. Herein, we report a simple, rapid, and sensitive method for the nucleic acid assay by fluorescence or naked eye using chemical cyclic amplification. The addition of hydroxylamine (HA) during the Fenton reaction can continuously generate hydroxyl radicals (•OH) via Fe3+/Fe2+ cycle, termed as "hydroxylamine boosts the Fenton reaction (Fenton-HA system)". Meanwhile, the reducing substances, such as terephthalic acid or o-phenylenediamine, react with •OH to generate oxidized substances that can be recognized by the naked eye or detected by fluorescence so as to realize the detection of Fe3+. The concentration of Fe3+ has a good linear relationship with fluorescence intensity in the range of 0.1 to 100 nM, and the limit of detection is calculated to be 0.03 nM (S/N = 3). Subsequently, Fe was introduced into the nucleic acid hybridization system after the Fe source was transformed into Fe3+, and the nucleic acids were indirectly determined by this method. This Fenton-HA system was used for sensing HIV-DNA and miRNA-21 to verify the validity of this method in nucleic acid detection. The detection limits were as low as 2.5 pM for HIV-DNA and 3 pM for miRNA-21. We believe that our work has unlocked an efficient signal amplification strategy, which is expected to develop a new generation of highly sensitive chemical biosensors.

Bioaccumulation of organophosphorus flame retardants in marine organisms in Liaodong Bay and their potential ecological risks based on species sensitivity distribution

Although organophosphorus flame retardants (OPFRs) in aquatic environments have received increasing concern, little information is available on their bioaccumulation and trophic transfer in marine food webs. Consequently, the risks of OPFRs to marine ecosystems are unknown. In this study, seven OPFR compounds in marine biological samples collected from Liaodong Bay, Bohai Sea, were analyzed to evaluate their level and biological amplification effect in the marine food web. The total OPFRs of marine organisms in Liaodong Bay ranged from 2.60 to 776 ng/g ww, and lipids were critical factors affecting the concentration of OPFRs in marine species. Tris (2-ethylhexyl) phosphate (TEHP) and tris(1-chloro-2-propyl) phosphate (TCIPP) were the OPFRs most frequently detected in marine species. Still, tris(2-chloroethyl) phosphate (TCEP) was dominant in most marine species (16/24), and the content of chlorinated OPFRs was highest. At the same time, alkyl OPFRs and aryl OPFRs accounted for the same proportion. No correlation between OPFR concentration and the trophic level was observed in marine organisms from Liaodong Bay. It was shown in the results of the species sensitivity distribution that TCIPP in Chinese seawater does not pose a potential ecological risk to marine species. However, much work remains to be done on accumulating information and the ecological risks of OPFRs in different marine food webs.

An Ecological Approach to Control Pathogens of Lycopersicon esculentum L. by Slow Release of Mancozeb from Biopolymeric Conjugated Nanoparticles.

To control insects, weeds, and infections in crops, old-fashioned pesticide formulations (with massive quantities of heavy metals and a variety of chemicals) are used. By biological amplification via the food chain, many of these established pesticide formulations have accumulated in living systems and caused environmental pollution. To form a nanoparticulate matrix with a diameter ranging from 322.2 ± 0.9 to 403.7 ± 0.7 nm, mancozeb was embedded in chitosan-gum acacia (CSGA) biopolymers and loadings were confirmed via TEM and FTIR. Differential scanning calorimetry analyses were carried out as part of the investigation. Inhibition of Alternaria alternata by nanoparticles (NPs) with 1.0 mg/mL mancozeb (CSGA-1.0) was 85.2 ± 0.7 % at 0.5 ppm, whereas for Stemphylium lycopersici it was 62.1 ± 0.7% in the mycelium inhibition method. NPs demonstrated antimicrobial action in pot house environments. After ten hours, the mancozeb was liberated from the nanoformulations due to polymer matrix diffusion and relaxation, compared to 2 h for commercial mancozeb. Even while drug-loaded conjugated nanoparticles have equivalent antifungal activities, they have a lower release rate and, hence, reduced toxicology compared to commercial mancozeb. Therefore, this method can be employed to implement sustainable farming techniques in the future.

Trends and Health Risk of Trace Metals in Fishes in Liaodong Bay, China, From 2015 to 2020

Due to their toxicity, non-biodegradability, and biological amplification in the food chain, heavy metal accumulation in humans via the consumption of fishes has become a problem. In this study, we analyzed the concentrations of six trace metals (Cd, Cr, Cu, Ni, Pb, Zn) in 12 marine fish species collected from Liaodong Bay, China, from 2015 to 2020, to understand the pollution status of the bay and evaluate the impact of fish consumption on human health. In addition, 5 fish species with the potential to serve as bioindicators of metal contamination were identified. In general, the average concentrations were Zn > Pb ≈ Cr > Ni ≈ Cu > Cd. There were significant differences in the concentrations in the muscle tissues of fish with different feeding strategies. The total target hazard quotient values of some fish were greater than 1 in 2015, suggesting that people were susceptible to high health risks. The year 2017 had the lowest values, which suggests that metal pollution in Liaodong Bay may have been decreasing at that time. Monitoring of metal concentrations in marine fish from Liaodong Bay warrants more study.

Visual detection of aflatoxin B1 based on specific aptamer recognition combining with triple amplification strategy

A highly sensitive and specific visual detection method for aflatoxin B1 (AFB1) based on the target specificity of aptamer, rolling circle amplification (RCA) and enzyme catalysis biological amplification effect has been established. In this work, AFB1 aptamer immobilized on the surface of magnetic beads (MB) serves as a molecular recognition probe. In the absence of AFB1, the aptamer and auxiliary linking probe (LP) maintain a double stranded state due to partial base pair complementarities. By contrast, in the presence of AFB1, the aptamer preferentially binds to AFB1 specifically, and the LP later restores to a single stranded state. Subsequently, the RCA reaction is triggered by above-mentioned single stranded LP to generate long DNA strands, which are employed to capture amounts of signal probes (SP) and horse radish peroxidases (HRP). Finally, amounts of HRP catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) by H2O2 and leads to a dramatic color change of the solution from colorlessness to deep blue as a signal indicator, obtaining a high sensitivity, high specificity and visual detection of AFB1. Under optimal conditions, a good linear detection range (0.5–40 pg·mL−1) was achieved, and the limit of detection (LOD) was 0.13 pg·mL−1. Besides, the proposed aptasensor showed excellent specificity for AFB1 compared with five other mycotoxins. More than that, all reactions occur on the surface of the magnetic beads, which not only facilitates the detection operation process including the efficient isolation and collection of AFB1 from sample matrix, but also gets better selectivity and stronger resistibility to target analyte in complex sample matrix, adequately indicating its potential application in AFB1 practical detection.

Efficacy of Phytochemicals Against Mosquito Larvae: An Update to Integrated Mosquito Management

Mosquitoes are considered a most common human enemy; causes mortality, morbidity, monetary and social disruption. It plays a significant role in endemic conditions for Malaria, Japanese encephalitis (JE), Dengue, Yellow fever, Chikungunya, and Filariasis etc. The prolonged use of synthetic insecticides raises the risk of unintended harmful or fatal consequences such as biological amplification, mosquito resistance development, and negative impacts on environmental quality and non-target species, including human health. So that there is urgent need to find natural and environmentally safe phytochemicals as an alternative to synthetic pesticides from indigenous plant sources that are accessible to the whole local community. Phytochemicals are botanicals that are naturally occurring pesticides produced from floral resources, and they are a viable alternative to synthetic insecticides in Integrated Mosquito Management (IMM). Plant based insecticides are less toxic, delay the development of resistance because of its structure and easily biodegradable. In this article, we have summarized the recent studies published in 2020 and 2021 on the biopesticide effect of botanicals with their suitable solvents which possess strong potential against mosquitoes larva. We also focused on their mode of action on the target population, variance in larvicidal efficacy among Culex, Aedes, and Anopheles mosquitoes, polarity of solvents employed during extraction, active component, and prospective improvements in biological control.

Carbon Materials Advancing Microorganisms in Driving Soil Organic Carbon Regulation.

Carbon emission from soil is not only one of the major sources of greenhouse gases but also threatens biological diversity, agricultural productivity, and food security. Regulation and control of the soil carbon pool are political practices in many countries around the globe. Carbon pool management in engineering sense is much bigger and beyond laws and monitoring, as it has to contain proactive elements to restore active carbon. Biogeochemistry teaches us that soil microorganisms are crucial to manage the carbon content effectively. Adding carbon materials to soil is thereby not directly sequestration, as interaction of appropriately designed materials with the soil microbiome can result in both: metabolization and thereby nonsustainable use of the added carbon, or—more favorably—a biological amplification of human efforts and sequestration of extra CO2 by microbial growth. We review here potential approaches to govern soil carbon, with a special focus set on the emerging practice of adding manufactured carbon materials to control soil carbon and its biological dynamics. Notably, research on so-called “biochar” is already relatively mature, while the role of artificial humic substance (A-HS) in microbial carbon sequestration is still in the developing stage. However, it is shown that the preparation and application of A-HS are large biological levers, as they directly interact with the environment and community building of the biological soil system. We believe that A-HS can play a central role in stabilizing carbon pools in soil.

Touch DNA recovery from vehicle surfaces using different swabs.

Several methods of DNA collection are used in places or objects related to crimes, the most common being the use of swabs. However, it is known that the efficacy of touch DNA recovery can be affected by collection devices and surfaces. The aim of this work was to evaluate the efficiency of three different types of swab in recovering touch DNA collected from different parts of a vehicle. The following swabs were tested: PurFlock® swab (Puritan, USA), 4N6FLOQSwabs™ (Copan S.p.A., Italy), and cotton swab (Labor Import). The experiments were carried out in the same vehicle, using the gearshift knob, the parking brake lever, and the steering wheel as support for the collection of touch DNA. Swabs showed significant differences in the amount of DNA recovered (Hc = 53.52; p < 0.05) and in the rate of allele amplification (Hc = 24.3; p < 0.05). The results indicated a greater DNA recovery efficiency by PurFlock® swab, followed by cotton, and then 4N6FLOQSwabs™. However, there was no significant difference among the surfaces analyzed. PurFlock® swab was more efficient for recovering donor alleles than the others (cotton and 4N6FLOQSwabs™), especially for small DNA amounts. This swab was, therefore, suitable for collections in vehicles involved in crime. Furthermore, this study highlights the need to assess different materials and methods of collection of biological samples, considering collection, extraction, and amplification.

A universal array platform for ultrasensitive, high-throughput and microvolume detection of heavy metal, nucleic acid and bacteria based on photonic crystals combined with DNA nanomachine

The development of a universal, sensitive, and rapid assay platform to achieve detections of heavy metal, nucleic acid and bacteria is of great significance but it also faces a thorny challenge. Herein, a novel and universal array platform was developed by combining photonic crystals (PCs) and DNA nanomachine. The developed array platform integrated the physical and biological signal amplification ability of PCs and DNA nanomachine, resulting in ultrasensitive detections of Hg2+, DNA, and Shigella sonnei with limits of detection (LODs) of 22.1 ppt, 31.6 fM, and 9 CFU/mL, respectively. More importantly, by utilizing a microplate reader as signal output device, the array achieved high-throughput scanning (96 samples/3 min) with only 2 μL loading sample, which is advantageous for the detection of infectious dangerous targets. In addition, the PCs array could be obtained easily and rapidly based on self-assembly of colloidal nanospheres, and the DNA nanomachine was operated with enzyme-free and time-saving features. Benefiting from these merits, the proposed PCs array offered a powerful universal platform for large-scale detection of various analytes in the fields of pollution monitoring, epidemic control, and public health.

Roadmap on Universal Photonic Biosensors for Real-Time Detection of Emerging Pathogens

The COVID-19 pandemic has made it abundantly clear that the state-of-the-art biosensors may not be adequate for providing a tool for rapid mass testing and population screening in response to newly emerging pathogens. The main limitations of the conventional techniques are their dependency on virus-specific receptors and reagents that need to be custom-developed for each recently-emerged pathogen, the time required for this development as well as for sample preparation and detection, the need for biological amplification, which can increase false positive outcomes, and the cost and size of the necessary equipment. Thus, new platform technologies that can be readily modified as soon as new pathogens are detected, sequenced, and characterized are needed to enable rapid deployment and mass distribution of biosensors. This need can be addressed by the development of adaptive, multiplexed, and affordable sensing technologies that can avoid the conventional biological amplification step, make use of the optical and/or electrical signal amplification, and shorten both the preliminary development and the point-of-care testing time frames. We provide a comparative review of the existing and emergent photonic biosensing techniques by matching them to the above criteria and capabilities of preventing the spread of the next global pandemic.

The role of exosomes derived miRNAs in cancer.

Exosomes are 20-150nm cell secreting nano-bodies that helps in the transportation of various biomolecules, including micro ribonucleic acid (miRNA) in the human body during both normal and diseased conditions. The current review was planned to summarise the role of miRNA carried by circulatory exosomes in cancer. miRNA is responsible for contribution in cancer, regulation of gene expression, interfering in biological pathways, gene silencing or amplification, and also has a role in cancer resistance. (miRNA) plays a dynamic role in this process by regulating the genes related to drug resistance, cell proliferation, cell cycle and apoptosis through a tissue-specific fashion. Owing to its significances, micro ribonucleic acid has been reported to be the key regulator of cancer, metastasis and also a factor in cancer resistance, and is a better source of possible potential diagnostic biomarkers. Though many studies have explored the biological roles of RNAs in cancer, many facts are needed to be investigated for clinical applications.