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JOURNAL OF FLUORESCENCE · 2023 · IF 2.7

A ThT Derivative as Zn2+ Sensor Based on DNA G-quadruplex

Yuan Xinyu,Zhang Xiufeng,Hu Jinshan,Zhang Buyue,Ma Xiaoying,He Jinxing,Shi Lei

In this report, we developed a sensing strategy based on ThT-E (a ThT derivative) and DNA G-quadruplex for the label-free detection of Zn 2+ . In the absence of Zn 2+ , there was a fluorescence enhancement of ThT-E by interaction with human telomere sequence. On the addition of Zn 2+ , Zn 2+ induced a more compact antiparallel G-quadruplex to release ThT-E, resulting in fluorescence quenching. The detection limit was 0.6996 μM, and the fluorescence intensity showed a good linear relationship with the concentration of Zn 2+ in the range of 0-10 μM. This sensing strategy which only needs to mix two kinds of materials has the characteristics of label-feel, simple operation, short response time, economical and efficient.

CAS / MDL: 10196-18-6
研究领域: 分析化学 · 生物化学 · 材料科学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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ANALYST · 2022 ·Vol. 147 ·期 8 ·1631-1640 · IF 5.227

DNA hybridization-induced fluorescence variation in ThT: a new strategy of developing aqueous sensors for MO genes

Yunkang Ma,Chenhui Wu,Wenjie Yang,Zhigang Gao,Lihua Chen

Mycoplasma ovipneumoniae (MO), a disease-causing pathogen with some of the highest levels of morbidity and mortality, can spread silently at the herd level. A novel alternative nanoprobe for MO was prepared using porous metal–organic frameworks (MOFs) as the scaffold and hairpin DNA with specific sequences of MO and G4 segments as the probe. This preparation was based on the strong fluorescence emission by ThT (thioflavin T) in the limitation of G-quadruplexes with a cavity structure. The use of MOFs effectively limited the folding behavior of G4 as a part of the probe to improve the defect of the strong background signal caused by the free-state G4Probe in a buffer. The results from the selectivity experiment showed that only a trace amount of the target with lower ΔG could be the “key” to the highly efficient triggering of the release behavior of the G4Probe from MOFs and the subsequent change in the fluorescence behavior of ThT. The DNA targets could be determined by observing the change in the signal. More importantly, the probe showed a low detection limit and a good linear correlation between the concentration of target DNA ranging from 10−10 M to 10−6 M not only in buffer but also in natural complex media. Moreover, the operation involved in the whole strategy was simple and the total cost was low. These findings demonstrated the value of the probe in further clinical diagnosis. This study reports the successful construction of a ΔG-sensitive sensor for MO for the first time.

研究领域: 生物技术 · 微生物学 · 分析化学 · 纳米技术
✓ 已验证 DOI PubMed 高影响因子
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Analytical Methods · 2016 ·Vol. 8 ·期 48 ·8461-8465 · IF 1.915

Selection and characterization of thioflavin T aptamers for the development of light-up probes

Hongyan Wang,Jine Wang,Lijun Xu,Yajie Zhang,Yang Chen,Hongxia Chen,Renjun Pei

Aptamers are single-stranded oligonucleotides which are selected and are capable of binding certain targets tightly and specifically. Systematic Evolution of Ligands by Exponential Enrichment (SELEX) has been used to select aptamers for various targets such as proteins, metal ions, small molecules and so on. For the first time, we have used a modified SELEX technology to obtain DNA aptamers for a fluorogenic dye, thioflavin T (ThT). DNA aptamers with low micromolar dissociation constants were achieved after only 8 rounds of selection and the final minimized 33-mer aptamer showed 191-fold fluorescence enhancement in a saturated concentration. The truncated DNA aptamer showed excellent selectivity and specificity for ThT.

CAS / MDL: 2390-54-7
研究领域: 分子生物学 · 生物化学
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Forensic Chemistry · 2025 · IF 2.2

Forensic analysis of tetrahydrothiophene (THT) at gas explosion scenes: Adsorption-desorption dynamics on common substrates

Dong Han,Guan-Nan Zhang,Zhen-Wen Sun,Yao Liu

Tetrahydrothiophene (THT) is a common odorant in natural gas and plays a crucial role as key chemical evidence in post-explosion investigations. However, there has been limited research on the detection of THT in samples collected from gas explosion scenes. Headspace-solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) was employed to systematically investigate the adsorption and desorption behavior of THT on three common substrates: soil, sponge, and fabric. The results indicate that the physical and chemical properties of the substrate significantly influence the retention of THT. Polyurethane sponge and flax fiber exhibit the highest adsorption capacity, attributed to their larger specific surface area and porous structure. With soil as the representative substrate, humidity significantly reduces physical adsorption efficiency. Desorption kinetics exhibit significant temperature dependence: THT is released markedly faster as temperature rises, whereas storage under low-temperature conditions substantially prolongs its persistence on the substrate. The limit of detection (LOD) of the proposed method is 0.05   ng/g in soil, thus affording sufficient sensitivity for ultra-trace analysis. These findings provide a critical scientific basis for on-site evidence collection (prioritizing dry, porous materials), optimizing evidence preservation (immediate low-temperature storage), and enhancing the accuracy of forensic evidence interpretation in gas explosion cases.

CAS / MDL: 110-01-0
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ANALYTICAL SCIENCES · 2018 ·Vol. 34 ·期 2 ·149-153 · IF 1.355

A Thioflavin T-induced G-Quadruplex Fluorescent Biosensor for Target DNA Detection

Xiao Fang ZHANG,Hong Mei XU,Lei HAN,Nian Bing LI,Hong Qun LUO

The detection of disease-related DNA is of great significance for early and accurate diagnosis and therapy. In this work, we successfully achieved the sensitive detection of target DNA based on a thioflavin T (ThT)-induced G-quadruplex fluorescent biosensor. ThT, a water-soluble fluorescent dye, can induce G-rich sequences to form G-quadruplexes and obtain an obviously enhanced fluorescence. In this work, it was employed to construct a biosensor for the detection of HIV. When the target HIV existed, the hairpin DNA probes would be opened in succession and release the completely exposed G-rich sequence to combine with ThT. The simple and rapid biosensor performed satisfactory selectivity; it also exhibited sensitivity with a detection limit of 2.4 nM. With good performance in human serum, we believe that this optical biosensor has the potential to be applied to the practical detection of target DNA.

CAS / MDL: 77-86-1
研究领域: 生物化学 · 分子生物学 · 医学诊断
✓ 已验证 产品关联 DOI PubMed 高影响因子
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY · 2022 · IF 5.895

Lab-on-a-Molecule Probe: Multitarget Detection of Five Aromatic Pesticides Using a Supramolecular Probe under Single Wavelength Excitation

Kai-Ni Wei,Gui-Xian Song,Shu-Zhen Huang,Qing Tang,Jian-Hang Hu,Zhu Tao,Ying Huang

In order to prevent and control the effects of pesticide residues on human health and the ecological environment, the rapid, highly sensitive, and selective detection of multiple pesticide residues has become an urgent problem to be solved. Herein, a lab-on-a-molecule probe based on a host–guest complex (ThT@Q[8] probe) has been developed to simultaneously analyze multiple aromatic pesticides under single wavelength excitation, such as fuberidazole, thiabendazole, carbendazim, thidiazuron, and tricyclazole. The fluorescence titration spectra of the ThT@Q[8] probe with the five pesticides mentioned above showed that the fluorescence intensity exhibited a good linear correlation with the pesticide concentration and the limit of detection was as low as 10–7 M. Because the ThT@Q[8] probe exhibits diverse fluorescence color changes to the five pesticides studied under a 365 nm ultraviolet lamp, we fabricated a single probe used to detect multiple analytes in the RGB triple channel by extracting the RGB variations. Principal component analysis and linear discriminant analysis proved that the ThT@Q[8] probe can recognize and distinguish five pesticides and can be applied at different concentrations. In real samples, the ThT@Q[8] probe recognized and distinguished five pesticides in tap water and Huaxi River water. The 1H NMR spectra results proved that a charge-transfer complex of ThT and pesticides in the Q[8] cavity may be formed. Moreover, we selected a test strip as a carrier to detect pesticides. The results indicate it can be used to quickly and conveniently detect different pesticides due to the rapid color change. Besides, the ThT@Q[8] probe has good cell permeability and can be used to detect pesticide residues in living cells. This work has laid the foundation for the qualitative and quantitative multitarget detection of pesticide residues.

研究领域: 分析化学 · 环境科学 · 分子生物学
✓ 已验证 DOI 高影响因子
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MOLECULES · 2018 ·Vol. 23 ·期 10 ·2663 · IF 3.098

Design, Synthesis of N-phenethyl Cinnamide Derivatives and Their Biological Activities for the Treatment of Alzheimer’s Disease: Antioxidant, Beta-amyloid Disaggregating and Rescue Effects on Memory Loss

Tian Chai,Xiao-Bo Zhao,Wei-Feng Wang,Yin Qiang,Xiao-Yun Zhang,Jun-Li Yang

Gx-50 is a bioactive compound for the treatment of Alzheimer’s disease (AD) found in Sichuan pepper (Zanthoxylum bungeanum). In order to find a stronger anti-AD lead compound, 20 gx-50 (1–20) analogs have been designed and synthesized, and their molecular structures were determined based on nuclear magnetic resonance (NMR) and mass spectrometry (MS) analysis, as well as comparison with literature data. Compounds1–20were evaluated for their anti-AD potential by using DPPH radical scavenging assay for considering their anti-oxidant activity, thioflavin T (ThT) fluorescence assay for considering the inhibitory or disaggregate potency of Aβ, and transgenicDrosophilamodel assay for evaluating their rescue effect on memory loss. Finally, compound13was determined as a promising anti-AD candidate.Keywords:Alzheimer’s disease;gx-50;DPPH assay;ThT assay;transgenicDrosophilamodel assay

CAS / MDL: 19982-08-2
研究领域: 药理学 · 神经科学 · 化学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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TALANTA · 2015 · IF 3.545

New insights into side effect of solvents on the aggregation of human islet amyloid polypeptide 11–20

Yexuan Mao,Lanlan Yu,Ran Yang,Chuanguo Ma,Ling-bo Qu,Peter de B. Harrington

The formation of highly ordered fibrils for the human islet amyloid polypeptide (hIAPP) is considered as one of the precipitating factors of type 2 diabetes mellitus. In this study, an emerging new approach microscale thermophoresis and conventional ThT fluorescence assay were utilized to investigate the aggregation behavior of hIAPP 11–20 , giving a new insight of the solvent effect on the aggregation of hIAPP 11–20 . hIAPP 11–20 displayed different aggregation behaviors in various buffers, revealing that hIAPP 11–20 not only self-aggregates but also binds to solvent components. hIAPP 11–20 had a higher binding affinity for Tris than other selected buffers because multiple hydrogen bonds form, resulting in weaker self-aggregation of hIAPP 11–20 at the early stage of aggregation and prolonging the fibril formation process. hIAPP 11–20 displayed similar self-aggregation in both HEPES and pure water. Negatively charged phosphate ions in the PBS solution ‘neutralize’ the charges carried by hIAPP 11–20 itself to some extent, causing rapid aggregation of hIAPP 11–20 , and leading to a shorter fibrillation process of hIAPP 11–20 . These results revealed that solvents contribute to the aggregation of hIAPP 11–20 and demonstrated the affect of solvents on the activity of biomolecules. Additionally, as a new technique, microscale thermophoresis offers a powerful and promising approach to study the early stages of aggregation of peptides or proteins.

研究领域: 生物化学 · 分子生物学 · 糖尿病研究
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BIOSENSORS & BIOELECTRONICS · 2015 · IF 6.409

A label-free fluorescent biosensor for ultratrace detection of terbium (ш) based on structural conversion of G-quadruplex DNA mediated by ThT and terbium (ш)

Qiang Chen,Junfeng Zuo,Jinfeng Chen,Ping Tong,Xiujuan Mo,Lan Zhang,Jianrong Li

In this paper, a novel label-free fluorescent biosensor for terbium (ш) (Tb 3+ ) was proposed based on structural conversion of G-quadruplex DNA mediated by Thioflavin T (ThT) and Tb 3+ . In the presence of K + , ThT could bind to K + -stabilized parallel G-quadruplex, giving rise to high fluorescence intensity . Upon the addition of Tb 3+ , Tb 3+ could competitively bind to parallel G-quadruplex leading to the structural change, which resulted in fluorescence decrease. The change of fluorescence intensity (Δ F = F 0 – F ) showed a good linear response toward the concentration of Tb 3+ over the range from 1.0   pM to 10.0   µM with a limit of detection of 0.55   pM. This proposed biosensor was simple and cost-effective in design and in operation with ultrahigh sensitivity and selectivity . Thus, the proposed biosensor could be a promising candidate for monitoring ultratrace Tb 3+ in environment.

研究领域: 生物化学 · 环境科学
✓ 已验证 DOI 高影响因子
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MICROCHEMICAL JOURNAL · 2024 · IF 4.9

Redox-responsive degradation of DNA@MnO2 nanoprobe triggering DNAzyme recycling amplification for sensitive and label-free detection of glutathione

Xiuhua Zhang,Heqin Liu,Wanrong Hong,Yumin Wang,Jin-Wen Liu,Yingfeng Qin

In this work, a novel fluorescence nanoprobe based on MnO 2 nanosheets and DNAzyme cyclic amplification was developed for label-free detection of GSH. In our design, the DNA@MnO 2 nanoprobe was facilely prepared through the adsorption of DNA probes on the surface of MnO 2 nanosheets. Importantly, MnO 2 nanosheets in the DNA@MnO 2 nanoprobe can act as the recognizer of analytical target GSH. In the presence of GSH, the DNA@MnO 2 nanoprobe are decomposed owing to redox-responsive reaction between GSH and MnO 2 nanosheets, resulting in the generation of abundant Mn 2+ and the desorption of DNA probes. Subsequently, the Mn 2+ -dependent DNAzyme recycling amplification was rapidly initiated and a large number of the blocked G-rich sequences in DNA probes was released. The G-rich sequences were efficiently folded into G-quadruplex structures, which can remarkably light up the fluorescence intensity of thioflavin T (ThT). Experimental data demonstrated that the DNA@MnO 2 nanoprobes for GSH detection have a wide linear range from 0 to 2000 µM. Moreover, this fluorescence sensing strategy was successfully applied for the detection of GSH in complex biological media with satisfying results, providing a simple, facile and cost-efficient nanoplatform for GSH-related clinical disease diagnosis.

研究领域: 分析化学 · 纳米技术 · 生物医学工程 · 生物化学
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MICROCHIMICA ACTA · 2019 ·Vol. 186 ·期 3 ·1-8 · IF 5.479

Enzyme-free fluorometric assay for chloramphenicol based on double stirring bar-assisted dual signal amplification

Hong Feng,Lin Xiaoting,Wu Yongxiang,Dong Youren,Cao Yuting,Hu Futao,Gan Ning

An enzyme-free fluorometric assay is described that accomplishes dual signal amplification by making use of a two stirring bars. Two Y-shaped DNA probes were designed and placed on the bars. When the target (with chloramphenicol as model analyte) is added, it triggers target recycling and simultaneously catalyzes hairpin assembly (CHA). A large fraction of DNA primers is released by the analyte from the bar to the supernatant and open hairpins with G-quadruplex DNA sequence. The G-quadruplex can specifically bind thioflavin T (ThT) to emit fluorescence (with excitation/emission maxima at 445 and 485 nm) for quantification of chloramphenicol. An enzyme is not needed. ThT is added to the system as a fluorescent DNA probe. All this strongly reduces the cost for sensor construction and usage. The dual signal amplification steps occur simultaneously which reduces the detection time. The assay was successfully employed to the determination of CAP in spiked milk and fish samples within 60 min and with a 16 pM limit of detection (at S/ N  = 3). Schematic representation of a new method for the detection of chloramphenicol by using  two stirring bars. It is based on target recycling and catalyzed hairpin assembly amplification. CAP: chloramphenicol, ThT: thioflavin T, CHA: catalyzed hairpin assembly.

研究领域: 分析化学 · 生物化学 · 传感器与执行器
✓ 已验证 产品关联 DOI PubMed 高影响因子
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International Journal of Nanomedicine · 2024 · IF 6.6

Biosynthesis of Nanoparticles with Green Tea for Inhibition of β-Amyloid Fibrillation Coupled with Ligands Analysis

Mai Zhang,Yan Li,Chunli Han,Shiying Chu,Peng Yu,Wenbo Cheng

Background Inhibition of amyloid β protein fragment (Aβ) aggregation is considered to be one of the most effective strategies for the treatment of Alzheimer’s disease. (-)-Epigallocatechin-3-gallate (EGCG) has been found to be effective in this regard; however, owing to its low bioavailability, nanodelivery is recommended for practical applications. Compared to chemical reduction methods, biosynthesis avoids possible biotoxicity and cumbersome preparation processes.Materials and Methods The interaction between EGCG and Aβ42 was simulated by molecular docking, and green tea-conjugated gold nanoparticles (GT-Au NPs) and EGCG-Au NPs were synthesized using EGCG-enriched green tea and EGCG solutions, respectively. Surface active molecules of the particles were identified and analyzed using various liquid chromatography-tandem triple quadrupole mass spectrometry methods. ThT fluorescence assay, circular dichroism, and TEM were used to investigate the effect of synthesized particles on the inhibition of Aβ42 aggregation.Results EGCG as well as apigenin, quercetin, baicalin, and glutathione were identified as capping ligands stabilized on the surface of GT-Au NPs. They more or less inhibited Aβ42 aggregation or promoted fibril disaggregation, with EGCG being the most effective, which bound to Aβ42 through hydrogen bonding, hydrophobic interactions, etc. resulting in 39.86% and 88.50% inhibition of aggregation and disaggregation effects, respectively. EGCG-Au NPs were not as effective as free EGCG, whereas multiple thiols and polyphenols in green tea accelerated and optimized heavy metal detoxification. The synthesized GT-Au NPs conferred the efficacy of diverse ligands to the particles, with inhibition of aggregation and disaggregation effects of 54.69% and 88.75%, respectively, while increasing the yield, enhancing water solubility, and decreasing cost.Conclusion Biosynthesis of nanoparticles using green tea is a promising simple and economical drug-carrying approach to confer multiple pharmacophore molecules to Au NPs. This could be used to design new drug candidates to treat Alzheimer’s disease.

CAS / MDL: 16961-25-4 520-33-2
研究领域: 纳米技术 · 药理学 · 神经科学 · 材料科学 · 生物化学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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