Selenium-based electrocatalytic sensors for sensitive peroxynitrite detection in biological media: a bottom-up approach for functional interface design

用于生物介质中敏感过氧亚硝酸盐检测的硒基电催化传感器：功能界面设计的自下而上方法

• 批准号: 10799038
• 负责人: MEKKI BAYACHOU
• 金额: $ 10万
• 依托单位: CLEVELAND STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10799038
• 关键词: Administrative Supplement; Adult; Affect; Biochemical; Biocompatible Materials; Biological; Cardiovascular Diseases; Cell membrane; Cell physiology; Cells; Cessation of life; Chemicals; Chronic; Clinical; Collection; Coupled; DNA; Detection; Development; Devices; Disease; Electrodes; Electronics; Equipment; Film; Funding; Goals; Grant; Hot Spot; Image; Immune response; In Situ; Inflammation; Libraries; Link; Lipids; Liposomes; Maps; Measurement; Measures; Membrane Lipids; Methods; Microscope; Miniaturization; Modeling; Molecular; Monitor; Morbidity - disease rate; National Institute of General Medical Sciences; Natural graphite; Nitric Oxide; Oxidative Stress; Parents; Pathologic; Pathology; Performance; Peroxonitrite; Physiological; Physiological Processes; Process; Property; Proteins; Reaction; Reporting; Reproducibility; Research; Resolution; Role; Scanning; Scanning Probe Microscopes; Selenium; Sepsis; Signal Transduction; Source; Superoxides; Surface; System; Techniques; Testing; Thinness; Time; United States; United States National Institutes of Health; Vasodilation; Work; assault; biological systems; carbon fiber; cytotoxic; design; in vivo; instrument; interest; microsensor; miniaturize; miniaturized sensor; mortality; mortality statistics; oxidation; parent grant; sensor; stress reactivity; tool

Project Summary: Biological Background and the Challenge: Peroxynitrite (OONO-) is a potent cytotoxic compound that has been implicated in a host of pathophysiological conditions. Peroxynitrite is the primary product of the in vivo reaction of nitric oxide and superoxide anion-radical. The multifaceted physiologic reactions of this compound are directly implicated in a number of pathologies including, immune response, chronic inflammation, sepsis, and cardiovascular disease, to cite a few. just cardiovascular disease alone claims about 7 deaths every 4 minutes and sepsis affects 1.7 million adults in the United States each year and contributes to more than 250,000 deaths. The common thread that links peroxynitrite to all cited pathologies is its potent reactivity toward most cellular components including DNA, proteins, and lipids in cell membranes. Substantial oxidations and other transformations of proteins, DNA, and lipids contribute to the disruption of key cellular functions. The task of assessing peroxynitrite’s deleterious effects and examining hypotheses of its potential signaling roles is very difficult. The primary reason is that methods for measuring and monitoring accurately its concentration are inherently difficult due to low submicromolar concentrations under physiologic conditions coupled with its high reactivity. The electrochemical detection of peroxynitrite is a simpler and more convenient technique for application in biological settings. However, a systematic development of the right electrode interface that enhances the sensitivity and selectivity for this molecule is lacking. In the ongoing work of the parent grant we develop electrode interfaces decorated with organoselenides attached to the surface that serve as catalytic entities for sensitive and selective PON electrocatalytic determination. The request of this Administrative Supplement: Funds are requested to purchase an Electrochemical Scanning Probe Microscope (SECM) Equipment that is critical for the completion of aspects of specific Aims 2 and Specific Aim3 of the current NIH project. The equipment model is the ElProScan ELP electrochemical scanning microscope and related accessories. The SECM equipment is a unique scanning probe microscope system that uses micro- and ultramicrolectrodes as scanning probes in order to measure with high spatial resolution and accuracy local current measurement (activity/election exchange) over substrates of interest (in this case substrates in the form of liposomes of live single cells that generate peroxynitrite upon stimulation). The Request is to Support Work Well Within the Scope of the Current NIGMS-funded Grant: The requested equipment will be used in a manner to generate high-resolution topographic imaging in the form of electrochemical activity as a result of peroxynitrite release (see the accompanying 2-page Research Strategy). The instrument will be used in a variety of ways: 1) it will be used to test miniaturized sensors based on the most active selenides as developed in this work. 2) The equipment will also be used to test the most optimal sensors for PON measurement over live single cells. 3) We will also use the instrument over collections of cells. All of these ways will support aspects of Specific Aim2 and particularly Specific Aim3 as explained in Research Strategy section. The electrochemical scanning probe microscope that we request is compatible and will be mounted on our existing Olympus inverted microscope. This will allow us to easily and reproducibly validate the performance of our miniaturized selenide-based peroxynitrite sensors on biological sources of peroxynitrite. The setup will allow us to carry out scanning and current measurement at the same time and with high temporal and spatial resolution on biological materials such as live single cells and collections of cells as well lipid membranes of cell-like (NOS-Loaded Liposomes) as we described in our parent NIH project that is underway.

项目概要： 生物学背景和挑战：过氧亚硝酸盐 (OONO-) 是一种有效的细胞毒性化合物 过氧亚硝酸盐是多种病理生理状况的主要产物。 一氧化氮和超氧阴离子自由基的体内反应。 这种化合物直接涉及许多病理学，包括免疫反应、慢性病 炎症、败血症和心血管疾病，仅举几个心血管疾病的说法。 每 4 分钟约 7 人死亡，脓毒症每年影响美国 170 万成年人， 导致超过 250,000 人死亡的共同线索是将过氧亚硝酸盐与所有引用的内容联系起来。 其病理学特征在于它对大多数细胞成分（包括 DNA、蛋白质和脂质）具有强大的反应性。 蛋白质、DNA 和脂质的大量氧化和其他转化都有贡献。 破坏关键的细胞功能。 评估过氧亚硝酸盐的有害影响并检验其潜力的假设的任务 信号作用非常困难的主要原因是准确测量和监测的方法。 由于生理条件下亚微摩尔浓度较低，其浓度本质上是困难的 过氧亚硝酸盐的电化学检测更加简单且具有高反应性。 然而，在生物环境中应用更方便的技术还需要系统化的发展。 缺乏增强该分子灵敏度和选择性的正确电极界面。 在母公司资助的持续工作下，我们开发了用有机硒化物装饰的电极界面 到作为催化实体的表面，用于灵敏和选择性的 PON 电催化测定。 本行政补充的要求： 需要资金购买电化学扫描探针显微镜 (SECM) 设备 对于完成当前 NIH 项目的具体目标 2 和具体目标 3 的各个方面至关重要。 设备型号为ElProScan ELP电化学扫描显微镜及相关配件。 SECM 设备是一种独特的扫描探针显微镜系统，使用微和 超微电极作为扫描探针，以便以高空间分辨率和精度进行局部测量 对感兴趣的底物（在本例中为底物）的电流测量（活性/选举交换） 活单细胞的脂质体形式，在刺激时产生过氧亚硝酸盐）。 该请求旨在支持在当前 NIGMS 资助的拨款范围内做好工作： 所请求的设备将用于生成高分辨率地形成像 过氧亚硝酸盐释放导致的电化学活性形式（参见随附的 2 页 研究策略）。该仪器将以多种方式使用：1）它将用于测试小型化。 基于本工作中开发的最活跃的硒化物的传感器 2) 还将使用该设备。 通过实时单电池测试用于 PON 测量的最佳传感器 3) 我们还将使用 所有这些方法都将支持特定目标2的各个方面，尤其是细胞集合。 具体目标3如研究策略部分所述。 我们要求的电化学扫描探针显微镜是兼容的并将安装在我们的 现有的奥林巴斯倒置显微镜这将使我们能够轻松且可重复地验证。 我们的小型化硒基过氧亚硝酸盐传感器对生物来源的性能 该设置将使我们能够同时进行扫描和电流测量。 对生物材料（例如活的单细胞和集合）具有高时间和空间分辨率 正如我们在我们的母体 NIH 中所描述的，细胞以及细胞样脂质膜（NOS 负载脂质体） 正在进行的项目。

项目成果

Inducible Nitric Oxide Synthase Embedded in Alginate/Polyethyleneimine Hydrogel as a New Platform to Explore NO-Driven Modulation of Biological Function.

嵌入海藻酸盐/聚乙烯亚胺水凝胶中的诱导型一氧化氮合酶作为探索 NO 驱动的生物功能调节的新平台。

• 发表时间: 2023-02-07
• 作者: Maher, Shaimaa;Smith, Lauren A;El;Kalil, Haitham;Sossey;Bayachou, Mekki
• 通讯作者: Bayachou, Mekki

Microspheres with 2D rGO/Alginate Matrix for Unusual Prolonged Release of Cefotaxime.

具有 2D rGO/藻酸盐基质的微球可实现头孢噻肟异常长时间释放。

• 发表时间: 2023-05-01
• 作者: Gomaa, Islam;Emam, Merna H;Wassel, Ahmed R;Ashraf, Kholoud;Hussan, Sara;Kalil, Haitham;Bayachou, Mekki;Ibrahim, Medhat A
• 通讯作者: Ibrahim, Medhat A