ADVANCED FUNCTIONAL MATERIALS
·
2024
·
IF 18.5
Zhenzhen Xu,Zhigang Wang,Dingmeng Hu,Huiqin Chen,Yufei Yan,Yuan Li,Mengjing Tu,Qiaofeng Shen,Xuanyi Liu,Rui Li,Cheng Lu,Fei Xue,Congying Xie,Gen Yang
5-Hydroxymethylcytosine in genomic DNA (5hmC-DNA), a predominant epigenetic mark, plays key roles in a broad range of bioprocesses such as cancer progression. However, efficient detection of 5hmC-DNA, especially from highly complex biological specimens, remains a harsh challenge due to its low abundance as well as the serious disturbance of non-targets. Leveraging MXene as the signal booster, an electrochemical biosensing technique is presented here for ultrasensitive detection of 5hmC-DNA from complex biological samples. The developed technique integrates enzyme-catalyzed transform of the hydroxyl group of 5hmC, sequence-specific recognition of the 5hmC-DNA targets, and facile electrochemical reaction. Remarkably, nanosized Ti 3 C 2 T x MXene is introduced as a highly conductive agent, managing to greatly boost the signals. Exhibiting outstanding consistency and robustness, the technique not only achieves excellent linear responses in a broad range of 5hmC-DNA (1.0 × 10 −13 –1.0 × 10 −9 m ) with the lowest detection limit of 59 fM but also possesses excellent selectivity against 5hmC analogs of including cytosine (C)- and 5-methylcytosine (5mC). More importantly, a proof-of-concept validation demonstrates the extraordinary ability of this technique for directly detecting 5hmC-DNA from complex backgrounds including cellular genomic extracts and serum of irradiated or non-irradiated mice, suggesting its highly promising potential for diverse applications such as early cancer screening.
研究领域:
分子生物学
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分析化学
·
医学
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DOI
高影响因子
JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY
·
2017
·
IF 2.673
Tian-Fu He,Zhi-Hong Zhang,Xin-An Zeng,Lang-Hong Wang,Charles S. Brennan
This work was aimed to investigate the antibacterial action of cinnamaldehyde (CIN) against Escherichia coli ATCC 8735 ( E. coli ) based on membrane fatty acid composition analysis, alterations of permeability and cell morphology as well as interaction with genomic DNA . Analysis of membrane fatty acids using gas chromatography–mass spectrometry (GC–MS) revealed that the proportion of unsaturated fatty acids (UFA) and saturated fatty acids (SFA) were the major fatty acids in plasmic membrane, and their levels were significantly changed after exposure of E. coli to CIN at low concentrations. For example, the proportion of UFA decreased from 39.97% to 20.98%, while the relative content of SFA increased from 50.14% to 67.80% as E. coli was grown in increasing concentrations of CIN (from 0 to 0.88 mM). Scanning electron microscopy (SEM) showed that the morphology of E. coli cells to be wrinkled, distorted and even lysed after exposure to CIN, which therefore decreased the cell viability . The binding of CIN to genomic DNA was probed using fluorescence, UV–Visible absorption spectra, circular dichroism, molecular modeling and atomic force microscopy (AFM). Results indicated that CIN likely bound to the minor groove of genomic DNA, and changed the secondary structure and morphology of this biomacromolecule. Therefore, CIN can be deem as a kind of natural antimicrobial agents, which influence both cell membrane and genomic DNA.
研究领域:
微生物学
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分子生物学
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药理学
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DOI
PubMed
高影响因子
Scientific Reports
·
2017
·Vol. 7
·期 1
·1-9
·
IF 4.259
Li Lanhui,Jin Mingliang,Sun Chenglong,Wang Xiaoxue,Xie Shuting,Zhou Guofu,van den Berg Albert,Eijkel Jan C. T.,Shui Lingling
DNA fragmentation down to a precise fragment size is important for biomedical applications, disease determination, gene therapy and shotgun sequencing. In this work, a cheap, easy to operate and high efficiency DNA fragmentation method is demonstrated based on hydrodynamic shearing in a bubbling system. We expect that hydrodynamic forces generated during the bubbling process shear the DNA molecules, extending and breaking them at the points where shearing forces are larger than the strength of the phosphate backbone. Factors of applied pressure, bubbling time and temperature have been investigated. Genomic DNA could be fragmented down to controllable 1–10 Kbp fragment lengths with a yield of 75.30–91.60%. We demonstrate that the ends of the genomic DNAs generated from hydrodynamic shearing can be ligated by T4 ligase and the fragmented DNAs can be used as templates for polymerase chain reaction. Therefore, in the bubbling system, DNAs could be hydrodynamically sheared to achieve smaller pieces in dsDNAs available for further processes. It could potentially serve as a DNA sample pretreatment technique in the future.
研究领域:
生物技术
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分子生物学
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DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
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2021
·
IF 4.952
Hui Li,Mao Zhang,Keren Agyekumwaa Addo,Yigang Yu,Xinglong Xiao
The antibacterial mechanisms of cuminaldehyde ( p -Isopropylbenzaldehyde, CUM) against Staphylococcus aureus ATCC 6538 ( S. aureus ) were investigated from the aspects of membrane damage, and interaction with genomic DNA. The results demonstrated that the MIC of CUM against S. aureus was 0.8 μL/mL. Approximately 6 log CFU/mL of S. aureus could be killed after treatment with MIC of CUM after 8 h. Additionally, CUM increased the permeability of the cell membrane by inducing visible changes in cell morphology. The results of gel electrophoresis, spectroscopic analysis, and molecule docking indicated that CUM could bind to DNA through groove mode and change the secondary structure of DNA, and ultimately lead to the inactivation of cells. Compared with untreated samples, application of CUM (2 or 4 MIC) on sauced beef caused an obvious decrease in S. aureus counts and effectively inhibited the growth of S. aureus . Moreover, CUM at a concentration of 2 MIC had no significant effect on the taste and aroma score of sauced beef. Therefore, CUM could play a dual antibacterial effect by influencing cell membrane and genomic DNA, and controlling the growth of S. aureus in the food industry.
研究领域:
食品科学
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微生物学
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DOI
高影响因子
EMBO REPORTS
·
2025
·
IF 6.2
LiJunlong,SongYihao,GuoXiao,HeZheng-Guo
With the increasing prevalence of drug-resistant bacteria, antimicrobial resistance emerges as a global public health threat. Mycobacteriophages show exciting prospects for the treatment of drug-resistant bacterial infections. However, the molecular mechanism through which they escape host bacterial defenses remains unclear. Here, we report that the gene gp48 of the mycobacteriophage A10ZJ24, which encodes a metallophosphoesterase-like protein, is essential for killing Mycobacterium tuberculosis. Gp48 is expressed during early stages of phage infection, and the Gp48 protein efficiently disrupts mycobacterial genomic DNA integrity, thereby silencing the expression of multiple anti-phage defense genes. While gp48-deletion phages infect and inject their DNA normally into M. tuberculosis cells, they are not able to impair the activation of the bacterial anti-phage genes which inhibit the replication of the genomic DNA of the mutant phage. This study thus identifies a phage metallophosphoesterase as a novel tool for subverting host bacterial antiviral immunity and killing M. tuberculosis. Our work fills a critical gap in the current knowledge on the arms race between mycobacteriophages and M. tuberculosis.
研究领域:
微生物学
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分子生物学
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免疫学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2016
·Vol. 409
·期 6
·1615-1625
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IF 3.125
Wang Lang-Hong,Zhang Zhi-Hong,Zeng Xin-An,Gong De-Ming,Wang Man-Sheng
Thymol (2-isopropyl-5-methylphenol) is a natural ingredient used as flavor or preservative agent in food products. The antibacterial mechanism of thymol against Gram-positive, Staphylococcus aureus was investigated in this work. A total of 15 membrane fatty acids were identified in S. aureus cells by gas chromatography–mass spectrometry. Exposure to thymol at low concentrations induced obvious alterations in membrane fatty acid composition, such as decreasing the proportion of branched 12-methyltetradecanoic acid and 14-methylhexadecanoic acid (from 22.4 and 17.3% to 7.9 and 10.3%, respectively). Membrane permeability assay and morphological image showed that thymol at higher concentrations disrupted S. aureus cell membrane integrity, which may decrease cell viability. Moreover, the interaction of thymol with genomic DNA was also investigated using multi-spectroscopic techniques, docking and atomic force microscopy. The results indicated that thymol bound to the minor groove of DNA with binding constant ( K
a ) value of (1.22 ± 0.14) × 10 4 M −1 , and this binding interaction induced a mild destabilization in the DNA secondary structure, and made DNA molecules to be aggregated.
Thymol exerts its antibacterial effect throught destruction of bacterial cell membrane and binding directly to genomic DNA
研究领域:
微生物学
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分子生物学
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药理学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
·
2023
·
IF 5.4
Yufang Li,Xiaozhao Tang,Zushun Yang,Jinze He,Ni Ma,Aixiang Huang,Yanan Shi
The combination of natural antimicrobial peptide BCp12/phenyllatic acid (BCp12/PLA) presents a more efficient antibacterial effect, but its antibacterial mechanism remains unclear. This study studied the synergistic antibacterial mechanism of BCp12 and PLA against S. aureus . The results demonstrated that the BCp12/PLA combination presented a synergistic antibacterial effect against S. aureus , with a fractional inhibitory concentration of 0.05. Furthermore, flow cytometry and scanning electron microscope analysis revealed that BCp12 and PLA synergistically promoted cell membrane disruption compared with the group treated only with one compound, inducing structural cell damage and cytoplasmic leakage. In addition, fluorescence spectroscopy analysis suggested that BCp12 and PLA synergistically influenced genomic DNA. BCp12 and PLA targeted enzymes related to peptidoglycan and DNA synthesis and interacted by hydrogen bonding and hydrophobic interactions with mur enzymes (murC, murD, murE, murF, and murG), dihydrofolate reductase, and DNA gyrase. Additionally, the combined treatment successfully inhibited microbial reproduction in the storage of pasteurized milk, indicating that the combination of BCp12 and PLA can be used as a new preservative strategy in food systems. Overall, this study could provide potential strategies for preventing and controlling foodborne pathogens.
研究领域:
微生物学
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食品科学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Integrative Biology
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2017
·Vol. 9
·期 10
·820-829
·
IF 3.252
Wang Lang-Hong,Wang Man-Sheng,Zeng Xin-An,Xu Xi-Ming,Brennan Charles S.
The antimicrobial mechanism of naringenin, one of the citrus antibacterial flavonoids against food-borne Staphylococcus aureus ATCC 6538, was investigated in this study. Analysis of gas chromatography-mass spectrometry (GC-MS) and fluorescence showed that relatively low concentrations of naringenin caused perturbations in the membrane fatty acid composition and the conformation of membrane proteins through changing the microenvironment of phenylalanine, tyrosine, and tryptophan residues. Exposure of naringenin at higher levels significantly increased membrane permeability and changed the morphology of S. aureus cells. The genomic DNA-binding of naringenin was also quantitatively monitored using UV-vis spectra in combination with multivariate curve resolution-alternating least squares (MCR-ALS) analysis, and the concentration and pure spectra profiles for the three reaction species (DNA, naringenin, and DNA–naringenin) were obtained. Moreover, the thermal behavior of DNA and docking studies revealed that naringenin preferentially bound to the A–T base pair regions of genomic DNA via groove binding, and atomic force microscopy and circular dichroism showed that naringenin induced mild secondary structure and obvious morphological variations of this biomacromolecule. These results suggested that naringenin exerting its antibacterial effects might be connected with disruption of the cytoplasmic membrane and DNA targeting effects in Staphylococcus aureus.
研究领域:
微生物学
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分子生物学
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食品科学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
·
2021
·Vol. 56
·期 11
·6039-6051
·
IF 3.713
Ruifei Wang,Meimei Fang,Xinyi Hu,Yigang Yu,Xinglong Xiao
Summary The development of effective natural antibacterial agents is important due to the insecurity of synthetic antimicrobial agents and consumer preferences. In this study, kojic acid (KA) and tea polyphenols (TP) were found to exhibit synergistic inhibitory effect against Escherichia coli O157:H7 with a fractional inhibitory concentration index of 0.25. KA combined with TP at 25% of their minimum inhibitory concentrations (MICs) completely inactivated E. coli O157:H7 within 4 h. Subsequently, propidium iodide uptake tests, genomic DNA interaction analysis, molecular docking, field emission scanning electron microscopy and biofilm formation assay were conducted to understand the synergistic antibacterial mechanism of KA-TP. The results demonstrated that KA-TP attacked the cell membrane cooperatively, thus disturbing membrane integrity and cell structure, and KA could bind to the genomic DNA to affect the biofilm formation of E. coli O157:H7. Moreover, the contamination of E. coli O157:H7 on raw salmon fillets was eliminated to varying degrees after soaking treatments with KA and/or TP for 30 min. The depuration effect was further enhanced when KA combined with TP, and more than 2-log reduction of E. coli O157:H7 was obtained under the treatment of KA-TP at 2 MIC, suggesting that KA-TP may serve as a natural compound disinfectant for ready-to-eat aquatic products.
研究领域:
食品科学
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微生物学
·
药理学
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DOI
高影响因子
Frontiers in Microbiology
·
2022
·
IF 6.064
Wang Meng,Deng Zifeng,Li Yanmei,Xu Keyong,Ma Yi,Yang Shang-Tian,Wang Jufang
Pseudomonas aeruginosa (P. aeruginosa), an opportunistic pathogen, is often associated with difficulties in treating hospital-acquired infections. Biofilms formed by P. aeruginosa significantly improve its resistance to antimicrobial agents, thereby, posing a great challenge to the combat of P. aeruginosa infection. Antimicrobial peptides (AMPs) have recently emerged as promising antibiofilm agents and increasingly attracting the attention of scientists worldwide. However, current knowledge of their antibiofilm behavior is limited and their underlying mechanism remains unclear. In this study, a novel AMP, named PEW300, with three-point mutations (E9H, D17K, and T33A) from Cecropin A was used to investigate its antibiofilm property and antibiofilm pathway against P. aeruginosa. PEW300 displayed strong antibacterial and antibiofilm activity against P. aeruginosa with no significant hemolysis or cytotoxicity to mouse erythrocyte and human embryonic kidney 293 cells. Besides, the antibiofilm pathway results showed that PEW300 preferentially dispersed the mature biofilm, leading to the biofilm-encapsulated bacteria exposure and death. Meanwhile, we also found that the extracellular DNA was a critical target of PEW300 against the mature biofilm of P. aeruginosa. In addition, multiple actions of PEW300 including destroying the cell membrane integrity, inducing high levels of intracellular reactive oxygen species, and interacting with genomic DNA were adopted to exert its antibacterial activity. Moreover, PEW300 could dramatically reduce the virulence of P. aeruginosa. Taken together, PEW300 might be served as a promising antibiofilm candidate to combat P. aeruginosa biofilms.
研究领域:
微生物学
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分子生物学
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传染病学
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药学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2017
·
IF 7.78
Wenjing Jiang,Linni Wu,Junling Duan,Huanshun Yin,Shiyun Ai
An ultrasensitive sandwiched electrochemiluminescence (ECL) immunosensor was developed for 5-hydroxymethylcytosine (5hmC) detection in genomic DNA by using Fe 3 O 4 @SiO 2 core–shell magnetic nanomaterial as a immobilization matrix for anti-5hmC antibody, PAMAM conjugated avidin and Ru(bpy) 2 (phen-5-NH 2 )(PF 6 ) 2 as signal amplification unit. Importantly, Fe 3 O 4 @SiO 2 nanoparticles were verified to not only possess enormous surface for loading antibody by amido link, but also exhibit excellent bioactivity . With the dual signal amplification strategy, the ECL immunosensor showed wide detection range from 0.1 to 30 nM with low detection limit of 0.047 nM (S/N = 3). Based on the specific immunoreaction, the developed method also illustrated excellent detection selectivity . The fabricated immunosensor was also applied to detect the 5hmC in genomic DNA of cancer tissue, which indicated that the immunosensor possess potential applications in clinical detection.
研究领域:
分析化学
·
生物化学
·
分子生物学
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2021
·Vol. 11
·期 6
·3324-3332
·
IF 3.361
Jia Xu,Dan Chen,Yuan Yang,Hongjian Gong,Wenqi Gao,Han Xiao
A simple and efficient approach for the rapid extraction of genomic DNA from blood using various amino-modified magnetic nanoparticles (AMNPs) has been described. The salmon sperm DNA was isolated from aqueous solution based on electrostatic interaction between the positively charged amino-groups of AMNPs and the negatively charged phosphate groups of the DNA. The results of ultraviolet-visible (UV-Vis) spectrometry showed that increasing number of amino groups on the AMNPs surface resulted in an improvement in DNA adsorption efficiency. Several variables including the extraction pH, adsorption time, ionic strength and quantity of AMNPs were optimized to achieve the best extraction efficiency with the proposed method. Acceptable adsorption efficiency of 92% and recovery of 91% were achieved using multi-amino modified MNPs (mAMNPs) with an extraction time of 10 min and an overall processing time of 30 min. The mAMNPs enabled genomic DNA capture from human whole blood, and the resulting mAMNP/DNA complexes could be directly used as templates for PCR amplification without the need for complex and time-consuming DNA elution and purification steps. Our results imply that this method can be used as an effective strategy for genomic DNA extraction and may be extended to other types of biological samples.
研究领域:
分子生物学
·
纳米技术
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生物技术
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）