BIOSENSORS & BIOELECTRONICS
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2024
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IF 10.7
Mei Zhao,Jingyi Guo,Zilin Chen,Fang Wang
5-formylcytosine (5 fC) and 5-carboxylcytosine (5caC) serve as key intermediates in DNA demethylation process with significant implications for gene regulation and disease progression . In this study, we introduce a novel electrochemical sensing platform specifically designed for the sensitive and selective detection of 5 fC and 5caC in DNA . Protein A-modified magnetic beads (ProtA-MBs) coupled with specific antibodies facilitate the immunorecognition and enrichment of these modified bases. Signal amplification is achieved through several chemical reactions involving the interaction between N 3 -kethonaxl and guanine , copper-free click chemistry for the attachment of dibenzocyclooctyne (DBCO)-Biotin, and the subsequent recognition by streptavidin-conjugated horseradish peroxidase (SA-HRP). The assay's readout is performed on a disposable laser-induced graphene (LIG) electrode, modified with the bead-antibody-DNA complex in a magnetic field , and analyzed using differential pulse voltammetry in a system employing hydroquinone (HQ) as the redox mediator and H 2 O 2 as the substrate. This immunosensor displayed excellent sensitivity, with detection limits of 14.8 fM for 5 fC across a 0.1–1000 pM linear range and 87.4 fM for 5caC across a 0.5–5000 pM linear range, and maintained high selectivity even in the presence of interferences from other DNA modifications . Successful application in quantifying 5 fC and 5caC in genomic DNA from cell extracts, with recovery rates between 97.7% to 102.9%, underscores its potential for clinical diagnostics. N 3 -kethoxal was used for the first time in an electrochemical sensor. This work not only broadens the toolkit for detecting DNA modifications but also provides a fresh impetus for the development of point-of-care testing (POCT) technologies.
研究领域:
分子生物学
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
NUCLEIC ACIDS RESEARCH
·
2019
·Vol. 47
·期 19
·e119
·
IF 11.147
Chen Danping,Wang Yang,Mo Mingming,Zhang Junjie,Zhang Yanfei,Xu Yuzhi,Liu Si-Yang,Chen Jun,Ma Yingjun,Zhang Li,Dai Zong,Cai Chun,Zou Xiaoyong
The current methods for quantifying genome-wide 5-methylcytosine (5mC) oxides are still scarce, mostly restricted with two limitations: assay sensitivity is seriously compromised with cost, assay time and sample input; epigenetic information is irreproducible during polymerase chain reaction (PCR) amplification without bisulfite pretreatment. Here, we propose a novel Polymerization Retardation Isothermal Amplification (PRIA) strategy to directly amplify the minute differences between epigenetic bases and others by arranging DNA polymerase to repetitively pass large electron-withdrawing groups tagged 5mC-oxides. We demonstrate that low abundant 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC) and 5-carboxycytosine (5caC) in genomic DNA can be accurately quantified within 10 h with 100 ng sample input on a laboratory real-time quantitative PCR instrument, and even multiple samples can be analyzed simultaneously in microplates. The global levels of 5hmC and 5fC in mouse and human brain tissues, rat hippocampal neuronal tissue, mouse kidney tissue and mouse embryonic stem cells were quantified and the observations not only confirm the widespread presence of 5hmC and 5fC but also indicate their significant variation in different tissues and cells. The strategy is easily performed in almost all research and medical laboratories, and would provide the potential capability to other candidate modifications in nucleotides.
研究领域:
分子生物学
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表观遗传学
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基因组学
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生物化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
ELECTROANALYSIS
·
2013
·Vol. 25
·期 7
·1697-1705
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IF 2.817
Xiaoxia Zheng,Lishi Wang
Direct electrocatalytic oxidation and simultaneous determination of 5-methylcytosine (5-mC) and cytosine(C) were realized in alkaline solutions by differential pulse voltammetry (DPV) based on an electrochemically reduced graphene oxide (er-GO) modified glassy carbon electrode (er-GO/GCE). The as-prepared er-GO/GCE exhibited good electrocatalytic activity towards the oxidation of 5-mC and C. Under optimum conditions, the er-GO/GCE was applied to the simultaneous determination of 5-mC and C with a significantly improved peak potential resolution (about 150 mV), and a linear relationship can be obtained in the range of 6–200.0 µmol/L and 8–250.0 µmol/L, respectively. In addition, the proposed method was further successfully applied to the analysis of methylation status in short CpG oligonucleotides with satisfactory results.
研究领域:
分析化学
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电化学
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生物化学
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产品关联
DOI
高影响因子
ANALYST
·
2018
·Vol. 143
·期 9
·2051-2056
·
IF 3.864
Yin Yang,Guoqing Yang,Hongfei Chen,Hui Zhang,Jiu-Ju Feng,Chenxin Cai
The levels of 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) in DNA (5-mC-DNA and 5-hmC-DNA) are strongly correlated with cancer occurrence and development. The ability to distinguish and quantitatively detect them is important for cancer research. We have developed a hybridization chain reaction (HCR)-based electrochemical assay for the signal-amplified detection of the relative contents of 5-mC-DNA and 5-hmC-DNA. The DNA duplexes (containing 5-mC-DNA and 5-hmC-DNA with different percentages) were modified on a gold electrode. Electroactive [Ru(NH3)6]3+ (RuHex) was used as the signal reporter, because it binds to DNA double strands. The duplexes can be cleaved by MspJI endonuclease without HCR, and result in a small peak current. However, the cleavage can be blocked after the 5-hmC-DNA duplex is converted to β-glucosyl-5-hydroxymethylcytosine (β-glu-5-hmC) by T4 β-glucosyltransferase (T4 β-GT), and with the addition of helper DNA, a long double-helix DNA was formed through HCR. A significantly amplified peak current can be achieved due to the adsorption of numerous RuHex. The electrochemical signal of RuHex is correlated to the content of 5-hmC-DNA. Upon fixing the total quantity of 5-mC-DNA and 5-hmC-DNA on the electrode, the signals increase with the increase in the percentage of 5-hmC-DNA for the HCR. With this assay, a detection limit of 0.05% for 5-hmC-DNA was achieved.
研究领域:
生物化学
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分子生物学
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癌症研究
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电化学
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产品关联
DOI
PubMed
高影响因子
CRYSTENGCOMM
·
2022
·Vol. 24
·期 15
·2777-2790
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IF 3.756
Fan-Zhi Bu,Yue-Ming Yu,Yu-Li Shen,Lu Liu,Cui-Wei Yan,Zhi-Yong Wu,Yan-Tuan Li
The current research takes advantage of the low solubility and hepatoprotective activity of the phenolic acid nutrient vanillic acid (VAA) to reduce the solubility and dissolution rate of the antifungal drug 5-fluorocytosine (FCY) via a cocrystallization strategy, thus gaining new insights into surmounting the dose-limited hepatotoxicity caused by the rapid and almost complete absorption of FCY. An FCY-phenolic acid nutraceutical cocrystal, namely, FCY–VAA–H2O, was directionally self-assembled and structurally characterized. The single-crystal X-ray structural analysis showed that water and VAA molecules occupy the polar sites of FCY in situ through strong hydrogen-bonding interplays, establishing a key plane structure (KPS) with hydrophobicity, which down-regulates the solubility and dissolution rate of the cocrystal as compared with FCY itself. The conclusion is supported by systematic theoretical studies. Molecular electrostatic potential (MEP) calculations revealed a polarity decrease in KPS in the cocrystal relative to free FCY, in accordance with the polarity diminution induced by intensive hydrogen-bonding energy, through the reduced density gradient (RDG) analysis, implying an obvious potentiation in hydrophobicity. Thereby, the theoretical calculations achieved positive correlations with the experimental measurements.
研究领域:
药学
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分子生物学
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理论化学
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产品关联
DOI
高影响因子
Clinics
·
2025
·
IF 2.4
Shan Xu,Xuming Lin,Fengling Jia
Objective Retinoblastoma (RB) is a type of intraocular tumor in childhood with a high lethality rate. N4-acetylcytosine (ac 4 C) modification is known to regulate multiple cancers, which is mediated by the only known ac 4 C writer N-Acetyltransferase 10 (NAT10). In this study, the authors aimed to reveal the mechanism of RB progression regulated by ac 4 C modification. Method Phenotypically, dot blot assay and quantitative real-time PCR were used to detect the ac 4 C levels and NAT10 expression in clinical samples and RB cell lines. Then, NAT10 was knocked down to assess its effect on glycolysis. Mechanically, RNA immunoprecipitation assay, immunofluorescence assay, and dual luciferase report were used to explore the mRNA modified by NAT10-mediated ac 4 C modification. Mice xenograft model was used to determine the effect of NAT10 on tumor growth in vivo . Results The present results demonstrated that the levels of ac 4 C and NAT10 were increased in cancer tissues and RB cell lines. Furthermore, NAT10 knockdown inhibited the glycolysis in RB cell lines. Moreover, the authors revealed that NAT10 knockdown decreased the ac 4 C modification and mRNA stability of HK1, while the inhibition of glycolysis by NAT10 knockdown was reversed by HK1 overexpression. Finally, NAT10 knockdown relieved the growth of tumors in mice models. Conclusion The authors illustrated that NAT10 plays an important role in the progression of RB by regulating the ac 4 C modification on HK1 mRNA and affects its stability, which may provide a novel theoretical basis for the treatment of RB.
研究领域:
肿瘤学
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分子生物学
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医学遗传学
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产品关联
DOI
PubMed
高影响因子
POLYMER INTERNATIONAL
·
2023
·Vol. 73
·期 3
·213-222
·
IF 3.2
Wangping Qin,Ruike Zhang,Yan Fu,Jie Chang
In order to reduce environmental pollution and improve material safety, biobased flame-retardant polymers are gradually becoming a new trend. We synthesized a fully biobased flame retardant called PACY by reacting phytic acid (PA) and cytosine (CY) in water, and then mixed the flame retardant with biodegradable poly(lactic acid) (PLA) to obtain composites, characterizing the performance of the composites by microscopic morphology, thermal properties, flame-retardant performance and so on. The test results showed that the PLA composite with 2 wt% PACY (PLA/2PACY) achieved UL-94V-1 and the PLA composite with 4 wt% PACY (PLA/4PACY) had a limiting oxygen index of 23.6%. Compared to pure PLA, PLA/4PACY had good flame retardancy with 12% less peak heat release and 19% less total heat release. Analysis of the flame-retardant mechanism showed that PACY could promote the formation of a graphitized carbon layer after burning PLA, therefore enhancing the flame-retardant properties of PLA. Through a thermal stability study, it was found that the temperature required for PACY mass loss of 5% was 275 °C and the residual weight reached 38% at 750 °C, which indicated high thermal stability and residual carbon rate. The addition of PACY also improved the thermal conductivity of PLA, with that of PLA/4PACY being 3.84 times higher than that of pure PLA. In addition, the tensile strength, flexural modulus, maximum force during impact and maximum energy received during fracture of PLA/4PACY increased by 3.3%, 8.1%, 29.2% and 17.5% respectively over pure PLA, effectively improving the mechanical properties of PLA. Overall, a fully biobased flame retardant was synthesized for improving the fire resistance of PLA degradable polymers. © 2023 Society of Industrial Chemistry.
研究领域:
材料科学
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环境科学
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产品关联
DOI
高影响因子
ANALYST
·
2020
·Vol. 146
·期 2
·597-604
·
IF 3.978
Zhourui Xiao,Han Meng,Xuefei Qin,Xueqing Sang,Yun Zhang,Yali Yuan
Herein, a novel electrochemical biosensor was constructed for the highly efficient detection of silver ions. A porous platform constructed with functionalized gold nanoparticles (AuPP) was electropolymerized on the gold electrode surface. The obtained polymer, analogous to a metal–organic framework, was used as the sensing platform together with cytosine–Ag+–cytosine interaction for dual signal amplification. The scanning electron microscopy (SEM) image of the AuPP platform exhibited a porous structure and considerable binding sites for C-riched single stranded DNA, leading to predictable silver ion preconcentration. Based on this strategy, the biosensor showed that the peak current in differential pulse voltammetry rose linearly as the concentration of silver ion increased from 0.005 to 3 μM with a detection limit of 1.3 nM. In addition, in the presence of other metal ions, such as Pb2+, Mn2+, Ni2+, Co2+, Cu2+, Zn2+, Na+, Ca2+, and Cd2+, at the same concentration, the current signal remained almost unchanged, manifesting high selectivity for Ag+. This proposed sensor might exhibit a novel fabrication method for metal ion detection with the aid of multiple AuPP materials by designing ligands with different functional groups.
研究领域:
分析化学
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纳米技术
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电化学
·
生物传感器
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2012
·
IF 5.856
Yang Tang,Xiang-Dong Gao,Yinsheng Wang,Bi-Feng Yuan,Yu-Qi Feng
DNA methylation is one of the major epigenetic modifications and has been involved in a number of biological processes in mammalian cells. Yeast is widely used as a model organism for studying cell metabolism, cell cycle regulation, and signal transduction. However, it remains controversial whether methylated cytosine (5-methylcytosine, 5mC) exists in the yeast genome. In the current study, we developed a highly sensitive method based on gas chromatography/mass spectrometry (GC/MS) and systematically examined the incidence of 5mC in 19 yeast strains, which represent 16 yeast species. Our results showed that DNA methylation is widespread in yeast and the genome-wide DNA methylation of the studied yeast strains ranged from 0.014 to 0.364%, which were 1 to 2 orders of magnitude lower than that in mammalian cells (i.e., 3–8%). Furthermore, we found that the 5mC content in yeast varied considerably at different growth stages and DNA methylation inhibitor 5-azacytidine could induce a decrease in genome-wide DNA methylation as that in mammalian cells. The demonstration of the universal presence of DNA cytosine methylation in yeast constituted the first and essential step toward understanding the functions of this methylation in yeast.
研究领域:
分子生物学
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遗传学
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微生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR STRUCTURE
·
2022
·
IF 3.841
Fan-Zhi Bu,Yue-Ming Yu,Yu-Li Shen,Zhi-Yong Wu,Yan-Tuan Li
The present investigation aims to fully exploit the structural dominance and liver protective efficacy of phenolic acid nutraceutical ferulic acid (FUA) towards decreasing antifungal drug 5-fluorocytosine (FTS) in solubility and dissolution rate through a cocrystallization stratagem, thereby achieving some fire-new understandings for addressing the side effects generated by the fast and inappropriate absorption from FTS. Toward this end, the cocrystalline complex of FTS with FUA, viz . FTS-FUA, as the first cocrystal of FTS with phenolic acid nutrient of hydroxycinnamic acid subgroup is directionally assembled and fully characterized. The crystal structure determined by the single-crystal X-ray diffraction method demonstrates that FUA molecules in the lattice seize the polar sites in situ of FTS founded on the compact hydrogen-bond interaction, therefore leading to the formation of a rigid S-shaped module (RSM) with hydrophobic potentials, which induces a simultaneous down-regulation of the cocrystal in solubility and intrinsic dissolution rate versus raw drug FTS itself. The experimental outcomes are strongly verified by relevant theoretical investigations. The molecular electrostatic potential and the mutual penetration distance authenticate that the RSM's polarity in the cocrystal is lower than that of FTS per se, which complies with the abatement in polarity attributable to the reinforced hydrogen-bonds energy as per topology analysis with atoms in molecules theory, thus giving a remarkable improvement in hydrophobicity for the cocrystal. As a result, the current contribution not only provides a new crystallization form for FTS, but also exemplifies the feasibility of cocrystallization clue from integrated experimental with a systematic theoretical approach to overcome intractable adverse effects of the antifungal agent.
研究领域:
药物化学
·
晶体学
·
理论计算
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Jie Xiang,Junyi Zhang,Lei Liao,Bingying Jiang,Ruo Yuan,Yun Xiang
The sensitive detection of ten-eleven translocation (TET) dioxygenase is of significance for understanding the demethylation mechanism of 5-methylocytosine (5mC), which is responsible for a wide range of biological functions that can affect gene expression in eukaryotic species. Here, a non-label and sensitive fluorescence biosensing method for TET assay using TET1 as the model target molecule is established on the basis of target-triggered Mg 2+ -dependent DNAzyme and catalytic hairpin assembly (CHA)-mediated multiple signal amplification cascades. 5mC sites in the hairpin DNA probe are first oxidized by TET1 into 5-carboxycytosine, which are further reduced by pyridine borane into dihydrouracil , followed by its recognition and cleavage by the USER enzyme to liberate active DNAzyme and G-quadruplex sequences from the probe. The DNAzyme further cyclically cleaves the substrate hairpins to trigger subsequent CHA reaction and DNAzyme cleavage cycles for yielding many G-quadruplex strands. Thioflavin T dye then intercalates into G-quadruplexes to cause a magnificent increase of fluorescence for high sensitivity assay of TET1 with 47 fM detection limit. And, application of this method for TET1 monitoring in diluted serum has also been confirmed.
研究领域:
生物化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
POLYMER DEGRADATION AND STABILITY
·
2020
·
IF 4.032
Wei Tao,Xu Hu,Jinhao Sun,Lijun Qian,Juan Li
Two phosphorus-nitrogen flame retardants were synthesized by using chlodiphenylphosphine (CDP) or diphenylphosphinyl chloride (DPP) and cytosine (Cy), respectively. The products are marked as CCDP and CDPP, and characterized by using Fourier transform infrared spectra (FT-IR), 31 P and 1 H nuclear magnetic resonance and thermogravimetric analysis (TGA). The CCDP and CDPP were used to modify the flame retardancy of polyamide 6 (PA6). The results indicated that 6 wt% CCDP and CDPP both make PA6 achieve the UL-94 V-0 rating and improve its LOI value. The degradation behavior was studied by TG-IR, melt volume-flow rate, etc. The results revealed that both CCDP and CDPP promote the degradation of PA6 and improve its flame retardancy by melting away. The CCDP tends to work in the gas phase and the CDPP in the condensed phase. In addition, in order to reduce the effect of flame retardant on the mechanical properties of PA6, CDPP was coated with epoxy resin . The results showed that the flame retardancy decrease after coating. Finally, melamine cyanurate was blended with the CDPP coated by epoxy to adjust the comprehensive properties of PA6 composite, and excellent flame retardancy and mechanical properties are obtained.
研究领域:
材料科学
·
高分子化学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）