Chronic Intermittent Hypoxia and Hyperalgesic Priming - Administrative Supplement

慢性间歇性缺氧和痛觉过敏启动 - 行政补充

• 批准号: 10844191
• 负责人: NATHANIEL Aaron JESKE
• 金额: $ 1.6万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10844191
• 关键词: 18 year old; Abate; Administrative Supplement; Affect; Afferent Neurons; American; Anti-Inflammatory Agents; Applications Grants; Behavior; Behavioral; Biochemical; Biological Assay; Cell Separation; Chemicals; Chronic; Clinical; Clinical Research; Complement; Continuous Positive Airway Pressure; DNA; DNA Binding; Dangerousness; Densitometry; Development; Diagnosis; Environment; Extramural Activities; Film; Fixatives; Friends; Funding; Future; Gel; Genetic Screening; Health; Hypoxia; Image; Immune; In Vitro; Inflammatory; Knowledge; Laboratories; Life; Link; Location; Macrophage; Maintenance; Measures; Membrane; Mission; Modeling; Molecular Genetics; Mus; National Institute of Neurological Disorders and Stroke; Neuroimmune; Neuronal Plasticity; Neurons; Nociceptors; Obstructive Sleep Apnea; Oxidative Stress; Pain; Pathogenicity; Patients; Pattern; Peripheral; Persistent pain; Personal Satisfaction; Pharmacology Study; Pharmacotherapy; Phase; Phenotype; Physiological; Pilot Projects; Population; Proteins; Proteomics; Public Health; Pump; Quality of life; RNA; Recurrence; Reproducibility; Research; Rodent; Roentgen Rays; Role; Sampling; Scanning; Services; Signal Pathway; Sleep; Sleep Disorders; Sleep disturbances; System; Technology; Testing; Tissue Sample; Tissues; Transcript; Translating; United States National Institutes of Health; Up-Regulation; Visualization; Western Blotting; Work; X-Ray Film; chronic pain; clinical diagnosis; comorbidity; cost; digital; digital imaging; experimental study; health determinants; hypoxia inducible factor 1; image processing; imager; imaging system; improved; in vivo; in vivo imaging; inducible Cre; innovation; molecular marker; multidisciplinary; nanoGold; nanoparticle drug; new technology; nocturnal Hypoxemia; pain behavior; pain chronification; pain sensation; preclinical study; protein biomarkers; protein expression

Our current extramural funding, through NINDS, R01 NS128574, entitled Chronic Intermittent Hypoxia and Hyperalgesic Priming, incorporates multiple proteomic experiments that depend on the visualization and quantification of protein expression patterns across multiple in vitro and in vivo tissue samples. Furthermore, Western blotting experiments are utilized throughout the grant proposal to support key experiments that track peripheral neuron and macrophage protein marker expression profiles. Unfortunately, our main piece of machinery for processing and visualizing Western blot membranes has come under a state of irreversible disrepair, and we require immediate replacement to continue experiments proposed in the application. We currently house an 18-year-old MiniMedical Services X-ray developer and processor to visualize Western blot membranes. This developer requires monthly service to clean and replace reservoirs with environmentally- dangerous developer and fixative solutions to produce films that are then digitally scanned and quantified for band densitometry by NIH Image 1.62 to determine protein expression patterns. Several months ago, 2 of the three necessary reservoir pumps failed. The cost to replace the broken pumps are estimated by service technicians to cost more than a brand-new machine, thereby forcing us to discontinue its use in our laboratory for proteomic research. We have since solicited bids to determine how we might replace the broken developer with a chemiluminescence imager. We received several quotes from multiple companies and submit the attached quote for your consideration. The ThermoFisher Life Technologies iBright CL750 Digital Imaging System is an economical replacement for the outdated MiniMed X-ray developer. This imaging system allows for imaging of chemiluminescent Western blots, protein gels, and DNA gels, and supersedes the requirement for multi- instrumental image processing (X-ray film to digital scanner) by integrating the image and sample analysis in one machine. This is expected to significantly improve experimental rigor and reproducibility by reducing sampling variables that can be inadvertently introduced during handling between multiple machines. Furthermore, this technology is much more environmentally friendly by avoiding the need for dangerous chemical disposal. We respectfully submit this request for Administrative Supplemental Funding to continue NINDS-funded research with new technology that will reduce our environmental footprint and increase rigor and reproducibility to conduct work vital to the completion of the proposed studies.

我们目前的校外资金，通过NINDS，R 01 NS 128574，题为慢性间歇性缺氧和 痛觉过敏引发，结合了多个蛋白质组学实验，依赖于可视化和 在多个体外和体内组织样品中定量蛋白质表达模式。此外，委员会认为， 在整个拨款申请中，蛋白质印迹实验被用来支持跟踪 外周神经元和巨噬细胞蛋白标志物表达谱。不幸的是，我们的主要部分 用于处理和可视化蛋白质印迹膜的机器已经处于不可逆的状态， 年久失修，我们要求立即更换，以继续申请中提出的实验。 我们目前拥有一个18岁的MiniMedical Services X射线显影器和处理器，用于可视化Western 印迹膜。该开发商要求每月提供服务，以清洁和更换水库与环保- 危险的显影剂和固定剂溶液来生产胶片，然后对胶片进行数字扫描和量化， 通过NIH Image 1.62进行条带密度测定以确定蛋白质表达模式。几个月前，两名 三个必要的蓄水池水泵失灵了。更换损坏的泵的费用按服务估算 技术人员花费超过一个全新的机器，从而迫使我们停止在我们的实验室使用它 用于蛋白质组学研究。 我们已经征求投标，以确定我们如何可能取代破碎的显影液与化学发光 成像仪。我们收到了多家公司的报价，并提交了附件中的报价给您。 考虑. ThermoFisher Life Technologies iBright CL 750数字成像系统是一种经济型 替代过时的MiniMed X射线显影剂。该成像系统允许对 荧光免疫印迹，蛋白质凝胶和DNA凝胶，并取代了多- 仪器图像处理（X射线胶片到数字扫描仪），将图像和样品分析集成在 一台机器。预计这将通过减少 在多台机器之间的处理过程中可能无意中引入的采样变量。 此外，该技术通过避免对危险化学品的需要而更加环保 处置 我们恭敬地提交这份行政补充资金的请求，以继续NINDS资助的研究 新技术，将减少我们的环境足迹，并提高严格性和可重复性， 这对完成拟议的研究至关重要。