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Simultaneous detection of ovarian cancer related miRNA biomarkers with carboxylated graphene oxide modified electrochemical biosensor platform 利用羧基氧化石墨烯修饰的电化学生物传感器平台同时检测卵巢癌相关 miRNA 生物标记物
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-09-05 DOI: 10.1016/j.bioelechem.2024.108806

Ovarian cancer, known as “silent killer”, is a gynocological cancer with high mortality that usually diagnosed in the late stages. Gold standard immunoassay technique is evaluation of CA-125 levels which is not merely specific to ovarian cancer. Therefore, there is a need for sensitive determination of more specific biomarkers. miR-200 family is RNA nucleic acids that known to be upregulated in the presence of ovarian cancer. Since diagnosis based on a single biomarker is prone to generate misleading results, it is important to develop point-of-care systems that allow diagnosis of multiple miRNAs. Herein, an electrochemical nanobiosensor platform was developed for the multiplexed and simultaneous detection of miR-200c and miR-141. Biorecognition part was constitutued of methylene blue and ferrocene labeled hairpin DNA probes immobilized onto carboxylated graphene oxide modified pencil graphite electrodes. Their hybridization with miRNAs were examined upon “signal-off” approach using Square Wave Voltammetry. The platform demonstrated a linear detection range of 0.1 pM to 10 nM for both miR-141 and miR-200c, with low detection limits of 0.029 pM and 0.026 pM, respectively. We assume that the developed biosensor platform may pave the way in early diagnosis of the disease and the development of more effective treatment strategies.

卵巢癌被称为 "无声杀手",是一种死亡率很高的妇科癌症,通常在晚期才被诊断出来。金标准免疫测定技术是对 CA-125 水平进行评估,但这并不仅仅针对卵巢癌。miR-200 家族是一种 RNA 核酸,已知在卵巢癌发生时会上调。由于基于单一生物标志物的诊断容易产生误导性结果,因此开发可诊断多种 miRNA 的床旁系统非常重要。在此,我们开发了一种电化学纳米生物传感器平台,用于同时多重检测 miR-200c 和 miR-141。生物识别部分由固定在羧基氧化石墨烯修饰的铅笔石墨电极上的亚甲基蓝和二茂铁标记的发夹式 DNA 探针组成。探针与 miRNA 的杂交采用方波伏安法进行检测。该平台对 miR-141 和 miR-200c 的线性检测范围为 0.1 pM 至 10 nM,检测限分别为 0.029 pM 和 0.026 pM。我们认为,所开发的生物传感器平台可为疾病的早期诊断和开发更有效的治疗策略铺平道路。
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引用次数: 0
Unlocking the electrochemical performance of glassy carbon electrodes by surface engineered, sustainable chitosan membranes 通过表面工程化的可持续壳聚糖膜释放玻璃碳电极的电化学性能
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-09-03 DOI: 10.1016/j.bioelechem.2024.108804

Chitosan coatings, derived from crustacean shell waste, possess inherent biocompatibility and biodegradability, rendering them suitable for various biomedical and environmental applications, including electrochemical biosensing. Its amine and hydroxyl functional groups offer abundant sites for chemical modifications to boost the charge transfer kinetics and provide excellent adhesion, enabling the construction of robust electrode-coating interfaces for electroanalysis. This study explores the role of electrostatically-driven chemical interactions and crosslinking density originating from different chitosan (Cs) and glutaraldehyde (Ga) concentrations in this aspect. Studying anionic ([Fe(CN)6]3−/4−), neutral (FcDM0/+), and cationic ([Ru(NH3)6]2+/3+) redox probes highlights the influence of Coulombic interactions with chitosan chains containing positively-charged pathways, calculated by DFT analysis. Our study reveals how a proper Ch-to-Ga ratio has a superior influence on the cross-linking efficacy and resultant charge transfer kinetics, which is primarily boosted by up to 20× analyte preconcentration increase, due to electrostatically-driven migration of negatively charged ferrocyanide ions toward positively charged chitosan hydrogel. Notably the surface engineering approach allows for a two-orders of magnitude enhancement in [Fe(CN)6]4− limit of detection, from 0.1 µM for bare GCE down to even 0.2 nM upon an adequate hydrogel modification.

壳聚糖涂层提取自甲壳类动物的外壳废物,具有固有的生物相容性和生物可降解性,因此适用于各种生物医学和环境应用,包括电化学生物传感。其胺和羟基官能团提供了丰富的化学修饰位点,可促进电荷转移动力学并提供出色的附着力,从而构建用于电分析的坚固的电极涂层界面。本研究探讨了不同壳聚糖(Cs)和戊二醛(Ga)浓度所产生的静电驱动化学相互作用和交联密度在这方面的作用。通过对阴离子([Fe(CN)6]3-/4-)、中性(FcDM0/+)和阳离子([Ru(NH3)6]2+/3+)氧化还原探针的研究,凸显了通过 DFT 分析计算出的与含有正电荷途径的壳聚糖链之间库仑相互作用的影响。我们的研究揭示了适当的 Ch-Ga 比如何对交联效果和由此产生的电荷转移动力学产生卓越的影响,这主要是由于带负电荷的亚铁离子在静电驱动下向带正电荷的壳聚糖水凝胶迁移,从而使分析物的预浓度提高了 20 倍。值得注意的是,通过表面工程方法,[Fe(CN)6]4- 的检测限提高了两个数量级,从裸 GCE 的 0.1 µM 降低到适当修饰水凝胶后的 0.2 nM。
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引用次数: 0
Anti-tumor effects of nanosecond pulsed electric fields in a murine model of pancreatic cancer 纳秒脉冲电场在小鼠胰腺癌模型中的抗肿瘤作用
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-31 DOI: 10.1016/j.bioelechem.2024.108803

Nanosecond Pulsed Electric Fields (nsPEFs) treatment has demonstrated anti-tumor effects on various cancer cell lines. However, the use of this treatment in pancreatic cancer is limited. This study demonstrated that nsPEFs treatment effectively suppressed the proliferation and metastasis of pancreatic cancer cells, while also inducing DNA damage. Meanwhile, animal experiments have shown that nsPEFs effectively suppressed the growth of pancreatic cancer, even in cases where the tumor volume exceeded 500–600 mm3 at the initiation of treatment. Notably, a single treatment session was found to significantly inhibit tumor growth, while also showing no adverse effects on the main organs of the mice. RNA sequencing and bioinformatics revealed that seven key genes (CDK1, CENPA, UBE2C, CCNB2, PLK1, CCNA2, and CCNB14) were significantly correlated with the overall survival rate of patients with pancreatic cancer. Through the application of the competing endogenous RNA (ceRNA) hypothesis, two miRNAs (has-let-7b-5p and hsa-miR-193b-3p) and four lncRNAs (MIR4435-2HG, ZNF436-AS1, LINC01089, and MIR4435-2HG) were identified as significantly impacting the overall survival of pancreatic cancer patients. We have effectively developed an mRNA-miRNA-lncRNA network that has the potential to stimulate further investigation into the underlying mechanisms of nsPEFs on pancreatic cancer.

纳秒脉冲电场(nsPEFs)治疗已证明对多种癌细胞株有抗肿瘤作用。然而,这种疗法在胰腺癌中的应用还很有限。这项研究表明,nsPEFs 治疗能有效抑制胰腺癌细胞的增殖和转移,同时还能诱导 DNA 损伤。同时,动物实验表明,nsPEFs 能有效抑制胰腺癌的生长,即使治疗开始时肿瘤体积超过 500-600 立方毫米。值得注意的是,单次治疗就能显著抑制肿瘤生长,同时对小鼠的主要器官也没有不良影响。RNA测序和生物信息学发现,七个关键基因(CDK1、CENPA、UBE2C、CCNB2、PLK1、CCNA2和CCNB14)与胰腺癌患者的总生存率显著相关。通过应用竞争性内源性 RNA(ceRNA)假说,我们发现两个 miRNA(has-let-7b-5p 和 hsa-miR-193b-3p)和四个 lncRNA(MIR4435-2HG、ZNF436-AS1、LINC01089 和 MIR4435-2HG)对胰腺癌患者的总生存率有显著影响。我们有效地建立了一个 mRNA-miRNA-lncRNA 网络,该网络有可能激发人们进一步研究 nsPEFs 对胰腺癌的潜在影响机制。
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引用次数: 0
Selective detection of alpha synuclein amyloid fibrils by faradaic and non-faradaic electrochemical impedance spectroscopic approaches 通过法拉第和非法拉第电化学阻抗光谱法选择性检测α-突触核蛋白淀粉样纤维。
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-30 DOI: 10.1016/j.bioelechem.2024.108800

This study utilized faradaic and non-faradaic electrochemical impedance spectroscopy to detect alpha synuclein amyloid fibrils on gold interdigitated tetraelectrodes (AuIDTE), providing valuable insights into electrochemical reactions for clinical use. AuIDE was purchased, modified with zinc oxide for increased hydrophobicity. Functionalization was conducted with hexacyanidoferrate and carbonyldiimidazole. Faradaic electrochemical impedance spectroscopy has been extensively explored in clinical diagnostics and biomedical research, providing information on the performance and stability of electrochemical biosensors. This understanding can help develop more sensitive, selective, and reliable biosensing platforms for the detection of clinically relevant analytes like biomarkers, proteins, and nucleic acids. Non-faradaic electrochemical impedance spectroscopy measures the interfacial capacitance at the electrode–electrolyte interface, eliminating the need for redox-active species and simplifying experimental setups. It has practical implications in clinical settings, like real-time detection and monitoring of biomolecules and biomarkers by tracking changes in interfacial capacitance. The limit of detection (LOD) for normal alpha synuclein in faradaic mode is 2.39-fM, The LOD for aggregated alpha synuclein detection is 1.82-fM. The LOD for non-faradaic detection of normal alpha synuclein is 2.22-fM, and the LOD for nonfaradaic detection of aggregated alpha synuclein is 2.40-fM. The proposed EIS-based AuIDTEs sensor detects alpha synuclein amyloid fibrils and it is highly sensitive.

本研究利用法拉第和非法拉第电化学阻抗光谱法检测金穿插四电极(AuIDTE)上的α-突触核蛋白淀粉样纤维,为临床使用电化学反应提供有价值的见解。AuIDE 是购买的,用氧化锌修饰以增加疏水性。采用六氰基铁酸酯和羰基二咪唑进行了功能化。法拉第电化学阻抗光谱已在临床诊断和生物医学研究中得到广泛应用,为电化学生物传感器的性能和稳定性提供了信息。这种认识有助于开发灵敏度更高、选择性更强和更可靠的生物传感平台,用于检测生物标记物、蛋白质和核酸等临床相关分析物。非法拉第电化学阻抗光谱法测量电极-电解质界面的界面电容,无需氧化还原活性物种,简化了实验设置。它对临床具有实际意义,例如通过跟踪界面电容的变化来实时检测和监测生物分子和生物标记物。法拉第模式下正常α突触核蛋白的检测限(LOD)为 2.39-fM,聚集α突触核蛋白的检测限为 1.82-fM。非远缘检测正常α突触核蛋白的 LOD 为 2.22-fM,非远缘检测聚集α突触核蛋白的 LOD 为 2.40-fM。所提出的基于 EIS 的 AuIDTEs 传感器可检测α-突触核蛋白淀粉样纤维,而且灵敏度很高。
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引用次数: 0
Long-term changes in transmembrane voltage after electroporation are governed by the interplay between nonselective leak current and ion channel activation 电穿孔后跨膜电压的长期变化受非选择性泄漏电流和离子通道激活之间相互作用的影响。
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-30 DOI: 10.1016/j.bioelechem.2024.108802

Electroporation causes a temporal increase in cell membrane permeability and leads to prolonged changes in transmembrane voltage (TMV) in both excitable and non-excitable cells. However, the mechanisms of these TMV changes remain to be fully elucidated. To this end, we monitored TMV over 30 min after exposing two different cell lines to a single 100 µs electroporation pulse using the FLIPR Membrane Potential dye. In CHO-K1 cells, which express very low levels of endogenous ion channels, membrane depolarization following pulse exposure could be explained by nonselective leak current, which persists until the membrane reseals, enabling the cells to recover their resting TMV. In U-87 MG cells, which express many different ion channels, we unexpectedly observed membrane hyperpolarization following the initial depolarization phase, but only at 33 °C and not at 25 °C. We developed a theoretical model, supported by experiments with ion channel inhibitors, which indicated that hyperpolarization could largely be attributed to the activation of calcium-activated potassium channels. Ion channel activation, coupled with changes in TMV and intracellular calcium, participates in various physiological processes, including cell proliferation, differentiation, migration, and apoptosis. Therefore, our study suggests that ion channels could present a potential target for influencing the biological response after electroporation.

电穿孔会引起细胞膜通透性的时间性增加,并导致可兴奋细胞和不可兴奋细胞的跨膜电压(TMV)发生长期变化。然而,这些跨膜电压变化的机制仍有待全面阐明。为此,我们使用 FLIPR 膜电位染料对两种不同的细胞系进行单次 100 µs 电穿孔脉冲后的 30 分钟内的跨膜电压进行了监测。在表达内源性离子通道水平很低的 CHO-K1 细胞中,脉冲暴露后的膜去极化可以用非选择性漏电流来解释,这种漏电流一直持续到膜重新闭合,使细胞恢复其静息 TMV。在表达多种不同离子通道的 U-87 MG 细胞中,我们意外地观察到膜在初始去极化阶段后出现超极化,但只有在 33 ℃ 时才出现,25 ℃ 时则没有。我们建立了一个理论模型,并用离子通道抑制剂进行了实验,结果表明超极化在很大程度上可归因于钙激活钾通道的激活。离子通道的激活与 TMV 和细胞内钙的变化共同参与了各种生理过程,包括细胞增殖、分化、迁移和凋亡。因此,我们的研究表明,离子通道可能是影响电穿孔后生物反应的潜在靶点。
{"title":"Long-term changes in transmembrane voltage after electroporation are governed by the interplay between nonselective leak current and ion channel activation","authors":"","doi":"10.1016/j.bioelechem.2024.108802","DOIUrl":"10.1016/j.bioelechem.2024.108802","url":null,"abstract":"<div><p>Electroporation causes a temporal increase in cell membrane permeability and leads to prolonged changes in transmembrane voltage (TMV) in both excitable and non-excitable cells. However, the mechanisms of these TMV changes remain to be fully elucidated. To this end, we monitored TMV over 30 min after exposing two different cell lines to a single 100 µs electroporation pulse using the FLIPR Membrane Potential dye. In CHO-K1 cells, which express very low levels of endogenous ion channels, membrane depolarization following pulse exposure could be explained by nonselective leak current, which persists until the membrane reseals, enabling the cells to recover their resting TMV. In U-87 MG cells, which express many different ion channels, we unexpectedly observed membrane hyperpolarization following the initial depolarization phase, but only at 33 °C and not at 25 °C. We developed a theoretical model, supported by experiments with ion channel inhibitors, which indicated that hyperpolarization could largely be attributed to the activation of calcium-activated potassium channels. Ion channel activation, coupled with changes in TMV and intracellular calcium, participates in various physiological processes, including cell proliferation, differentiation, migration, and apoptosis. Therefore, our study suggests that ion channels could present a potential target for influencing the biological response after electroporation.</p></div>","PeriodicalId":252,"journal":{"name":"Bioelectrochemistry","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1567539424001646/pdfft?md5=9b60750e97ae48ebdbe7c9b36e0ee95d&pid=1-s2.0-S1567539424001646-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142144534","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Disposable Zirconium trisulfide-Reduced graphene oxide modified conducting thread based electrochemical biosensor for lung cancer diagnosis 基于电化学生物传感器的一次性三硫化锆-还原氧化石墨烯修饰导电线用于肺癌诊断
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-23 DOI: 10.1016/j.bioelechem.2024.108801

Flexible technology in sensors have received much attention in monitoring of human health through various physiological indicators. Thus, it drawn a lot of interest in the development of flexible substrate for the diagnosis of various diseases via analysis of analytes. Present work focusses on the development of ecofriendly, portable, flexible, conducting thread (Th) and used as smart substrate for fabrication of biosensor towards ultrasensitive detection of the lung cancer biomarker (cytoskeleton-associated protein 4; CKAP4). The zirconium trisulfide-reduced graphene oxide nanocomposite and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) modified cotton thread based biosensor was fabricated via dip coating method. Next, successive immobilization of monoclonal antibodies of CKAP4 (anti-CKAP4) and bovine serum albumin (BSA) was performed via drop cast approach using fabricated electrode [nZrS3@rGO/PEDOT:PSS/Th]. The response of fabricated electrode (BSA/anti-CKAP4/ZrS3@rGO/PEDOT:PSS/Th) was recorded electrochemically versus CKAP4 concentration via chronoamperometry (CA). The results showed wider linear detection range of 6.25-800 pg mL−1, excellent sensitivity of 85.2 µA[log(pg mL−1)]-1cm−2 with good stability up to 42 days. The response of fabricated biosensor was supported by investigating response of CKAP4 biomarker present in patients of lung cancer (concentration as determined through enzyme-linked immunosorbent assay) and obtained results exhibited excellent correlation with that of standard samples.

传感器中的柔性技术在通过各种生理指标监测人体健康方面备受关注。因此,人们对开发柔性基底以通过分析物诊断各种疾病产生了浓厚的兴趣。目前的工作重点是开发环保、便携、灵活的导电线(Th),并将其用作制造生物传感器的智能基底,以实现对肺癌生物标志物(细胞骨架相关蛋白 4;CKAP4)的超灵敏检测。通过浸涂法,制备了三硫化锆还原氧化石墨烯纳米复合材料和聚(3,4-亚乙二氧基噻吩):聚(苯乙烯磺酸)(PEDOT:PSS)改性棉线生物传感器。接着,通过滴注法,将 CKAP4(抗 CKAP4)和牛血清白蛋白(BSA)的单克隆抗体连续固定在制成的电极[nZrS3@rGO/PEDOT:PSS/Th]上。通过时标法(CA)记录了制备电极(BSA/抗 CKAP4/ZrS3@rGO/PEDOT:PSS/Th)的电化学响应与 CKAP4 浓度的关系。结果表明,该传感器的线性检测范围为 6.25-800 pg mL-1,灵敏度高达 85.2 µA[log(pg mL-1)]-1cm-2,稳定性良好,可持续 42 天。通过研究肺癌患者体内 CKAP4 生物标志物的反应(通过酶联免疫吸附测定法确定浓度),证实了所制造的生物传感器的反应,所获得的结果与标准样品的结果具有极好的相关性。
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引用次数: 0
Bioimpedance formalism: A new approach for accessing the health status of cell and tissues 生物阻抗形式主义:获取细胞和组织健康状况的新方法
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-19 DOI: 10.1016/j.bioelechem.2024.108799

This manuscript describes a novel methodology for studying relaxation dynamics in tissues and cells using characteristic frequency of bioimpedance spectroscopy measurements. The Bioimpedance Formalism allows for the simultaneous study of bioelectrical parameters in the frequency and time domains, providing insight into possible relaxation processes occurring in the tissue or cell of interest. Results from the Cole-Cole analysis showed no multiple relaxation processes associated with heterogeneity, with a visible age group separation in males compared with females. The study of the relaxation dynamic in the time domain revealed that the β parameter can be used to analyse the charge carriers in tissues, cells, or cancer cells, potentially leading to new diagnostic and therapeutic approaches for cancer and other diseases. Overall, this approach presents a promising area of research for gaining insights into the electrical properties of tissues and cells using bioimpedance methods.

本手稿介绍了一种利用生物阻抗光谱测量的特征频率研究组织和细胞弛豫动力学的新方法。生物阻抗形式主义允许同时研究频域和时域中的生物电参数,从而深入了解相关组织或细胞中可能发生的弛豫过程。科尔-科尔分析的结果表明,没有与异质性相关的多重弛豫过程,男性与女性相比有明显的年龄组差异。对时域弛豫动态的研究表明,β 参数可用于分析组织、细胞或癌细胞中的电荷载体,有可能为癌症和其他疾病带来新的诊断和治疗方法。总之,这种方法为利用生物阻抗方法深入了解组织和细胞的电特性提供了一个前景广阔的研究领域。
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引用次数: 0
Advancement in SARS-CoV-2 diagnosis: A new and stable electrochemical biosensor for genomic RNA detection SARS-CoV-2 诊断的进展:用于基因组 RNA 检测的新型稳定电化学生物传感器
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-17 DOI: 10.1016/j.bioelechem.2024.108798

Coronavirus disease (COVID-19) is caused by infection with the SARS-CoV-2 virus, having already caused more than seven million deaths worldwide. Conventional techniques for SARS-CoV-2 detection have limitations, as high cost, low specificity, and longer analysis time, among others. Biosensors emerge as a necessary alternative to overcome the difficulties of current diagnostics. This paper reports a sensor platform where silver-doped zinc oxide nanomaterial (Ag:ZnONp) was used onto carbon screen-printed electrode and ethidium bromide as indicator for development of a specific electrochemical genosensor for COVID-19. This genosensor demonstrated good linearity between the concentrations of 5.62 × 104–5.62 copies/mL and a detection limit of 5 copies/mL with gRNA in patient’s samples, with a response time within 30 min. Molecular modeling and morphological analysis are in agreement with obtained electrochemical results. Additional techniques such as AFM, SEM, and EIS were conducted to characterize the morphological and electrochemical properties of the biosensor’s surface. The biosensor was also capable of detecting the target presence in spiked samples and demonstrated a stability of 60 days, higher than other similar biosensors for SARS-CoV-2.

冠状病毒病(COVID-19)是由 SARS-CoV-2 病毒感染引起的,目前已造成全球 700 多万人死亡。传统的 SARS-CoV-2 检测技术存在成本高、特异性低、分析时间长等局限性。生物传感器的出现是克服现有诊断技术困难的必要选择。本文报告了一种传感器平台,即在碳丝网印刷电极上使用掺银氧化锌纳米材料(Ag:ZnONp),并以溴化乙锭作为指示剂,开发出一种针对 COVID-19 的特异性电化学基因传感器。该基因传感器在患者样本中 gRNA 的浓度为 5.62 × 104-5.62 拷贝/毫升和检测限为 5 拷贝/毫升之间表现出良好的线性关系,响应时间在 30 分钟之内。分子建模和形态分析与电化学结果一致。为了表征生物传感器表面的形态和电化学特性,还采用了原子力显微镜(AFM)、扫描电子显微镜(SEM)和电致发光显微镜(EIS)等其他技术。该生物传感器还能检测加标样品中目标物的存在,并显示出 60 天的稳定性,高于其他类似的 SARS-CoV-2 生物传感器。
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引用次数: 0
A sensitive tobramycin electrochemical aptasensor based on multiple signal amplification cascades 基于多重信号放大级联的灵敏妥布霉素电化学适配传感器
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-15 DOI: 10.1016/j.bioelechem.2024.108797

The residue of tobramycin, a broad spectrum antibiotic commonly used in animal husbandry, has evitable impact on human health, which may cause kidney damage, respiratory paralysis, neuromuscular blockade and cross-allergy in humans. Sensitive monitoring of tobramycin in animal-derived food products is therefore of great importance. Herein, a new aptamer electrochemical biosensor for sensing tobramycin with high sensitivity is demonstrated via exonuclease III (Exo III) and metal ion-dependent DNAzyme recycling and hybridization chain reaction (HCR) signal amplification cascades. Tobramycin analyte binds aptamer-containing hairpin probe to switch its conformation to expose the toehold sequence, which triggers Exo III-based catalytic digestion of the secondary hairpin to release many DNAzyme strands. The substrate hairpins immobilized on the Au electrode (AuE) are then cyclically cleaved by the DNAzymes to form ssDNAs, which further initiate HCR formation of lots of long methylene blue (MB)-tagged dsDNA polymers on the AuE. Subsequently electro-oxidation of these MB labels thus exhibit highly enhanced currents for sensing tobramycin within the 5–1000 nM concentration range with an impressive detection limit of 3.51 nM. Furthermore, this strategy has high selectivity for detecting tobramycin in milk and shows promising potential for detect other antibiotics for food safety monitoring.

妥布霉素是一种常用于畜牧业的广谱抗生素,其残留对人体健康有一定的影响,可能会对人体造成肾脏损害、呼吸麻痹、神经肌肉阻滞和交叉过敏。因此,对动物源性食品中托布霉素的灵敏监测至关重要。本文通过外切酶 III(Exo III)和金属离子依赖性 DNA 酶循环和杂交链反应(HCR)信号放大级联,展示了一种新型适配体电化学生物传感器,可高灵敏度地检测妥布霉素。妥布霉素分析物与含aptamer的发夹探针结合,改变其构象以暴露趾hold序列,从而触发基于Exo III的二级发夹催化消化,释放出许多DNA酶链。然后,固定在金电极(AuE)上的底物发夹会被 DNA 酶循环裂解,形成 ssDNA,从而进一步在 AuE 上启动 HCR,形成大量亚甲基蓝(MB)标记的长 dsDNA 聚合物。随后,这些 MB 标签发生电氧化,从而显示出高度增强的电流,可在 5-1000 nM 浓度范围内感测妥布霉素,检测限为 3.51 nM,令人印象深刻。此外,这种方法对检测牛奶中的妥布霉素具有很高的选择性,并有望用于检测食品安全监控中的其他抗生素。
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引用次数: 0
Respiration suppress and apoptosis-like cell death of Escherichia coli in direct current therapy mediated by polypyrrole conductive hydrogel 在聚吡咯导电水凝胶介导的直流治疗中抑制大肠杆菌的呼吸并使其细胞凋亡
IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-15 DOI: 10.1016/j.bioelechem.2024.108796

Targeting oxidative phosphorylation of bacteria is a novel antibiotic strategy leading to rapid cell death as a result of respiration suppress. Herein, a conductive polymer termed polypyrrole (PPy) is used to short-circuit the electron transfer chain (ETC) of bacteria cells owing to its higher electron affinity to electrons than all of the electron carriers on ETC. A hydrogel is fabricated using PPy which is anticipated to seize electrons from ETC and inhibit respiration of bacteria cells. The results show that the prepared PPy hydrogel can mediate an effective direct current (DC) antibacterial therapy which greatly enhances intracellular reactive oxygen species (ROS) level of Escherichia coli (E. coli), suppresses respiration, induces apoptosis-like cell death of E. coli accompanied by chromosomal condensation and loss of structural integrity, and rapidly cleared E. coli infection in vivo. Taken into the photothermal property of PPy, a combined direct current-photothermal therapy is developed which can enhance bacteria-killing effects with the assistance of an 808 nm laser. Our findings provide a new antibiotic strategy with metabolic pathway as a target.

以细菌的氧化磷酸化为靶标是一种新型抗生素策略,可抑制呼吸作用,导致细胞迅速死亡。在本文中,一种被称为聚吡咯(PPy)的导电聚合物被用于短路细菌细胞的电子传递链(ETC),因为它对电子的亲和力高于 ETC 上的所有电子载体。利用 PPy 制备的水凝胶有望从 ETC 中夺取电子并抑制细菌细胞的呼吸作用。结果表明,所制备的 PPy 水凝胶可作为一种有效的直流电(DC)抗菌疗法,大大提高大肠杆菌(E. coli)细胞内活性氧(ROS)水平,抑制呼吸,诱导大肠杆菌细胞凋亡样死亡,同时伴有染色体凝结和结构完整性丧失,并迅速清除体内大肠杆菌感染。考虑到 PPy 的光热特性,我们开发了一种直流-光热联合疗法,在 808 纳米激光的辅助下增强了杀灭细菌的效果。我们的研究结果提供了一种以代谢途径为靶点的新型抗生素策略。
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引用次数: 0
期刊
Bioelectrochemistry
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