ShEEP Request for Biotek Cytation 5 Imaging System

ShEEP 请求 Biotek Cytation 5 成像系统

• 批准号: 10175798
• 负责人: KARIN N. WESTLUND-HIGH
• 金额: --
• 依托单位: ALBUQUERQUE VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10175798
• 关键词: Acute Renal Failure with Renal Papillary Necrosis; Address; Americas; Analgesics; Anesthesiology; Anions; Antineoplastic Agents; Anxiety; Apoptosis; Autophagocytosis; Award; Bacterial Infections; Behavior; Biochemical; Biological Assay; Biological Markers; Blood Pressure; Blood Vessels; Brain; Brightfield Microscopy; Cancer cell line; Carrier Proteins; Cells; Chlorides; Chronic; Chronic Kidney Failure; Complex; Computer software; Critical Care; Cultured Cells; Data; Development; Diabetes Mellitus; Diagnostic; Disease; Distal; Diuretics; Doctor of Medicine; Doctor of Philosophy; Dose; Doxazosin; Drug Kinetics; Duct (organ) structure; End stage renal failure; Environment; Epithelial Cells; Equipment; Fluorescence; Fluorescence Microscopy; Fluorescence Polarization; Funding; Gene Expression Profile; Genetically Engineered Mouse; Goals; Growth; Health; Health Sciences; Healthcare Systems; Heart Diseases; Homeostasis; Hospitals; Human; Image; In Vitro; Incubators; Irritable Bowel Syndrome; Kidney; Kidney Diseases; Knockout Mice; Lead; Liquid substance; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Maximum Tolerated Dose; Measures; Medical; Medicine; Mental Depression; Milk Thistle; Milk thistle extract; Minor; Mitochondria; Modeling; Molecular; Monitor; Mus; Mycoses; Nephrology; Nephrons; Nerve; Neuraxis; New Mexico; Obesity; Oncology; Opioid; Oral Administration; Pain; Patients; Persian Gulf Syndrome; Pharmaceutical Preparations; Pharmacodynamics; Pharmacologic Substance; Physicians; Play; Post-Traumatic Stress Disorders; Pre-Clinical Model; Prediabetes syndrome; Procyanidins; Property; RNA; Rattus; Reader; Reporting; Research; Research Personnel; Robotics; Role; Safety; Savings; Scientist; Sheep; Silymarin; Sleep; Sodium Chloride; Standardization; Structure of parenchyma of lung; Systemic blood pressure; Technology; Testing; Time; Toxic effect; Translational Research; Treatment Efficacy; Trigeminal System; United States; Universities; Veterans; Veterans Health Administration; Water; Work; Xenograft Model; absorption; anti-cancer; arm; blood pressure regulation; bronchial epithelium; cancer therapy; cellular imaging; chronic pain; constriction; design; dietary salt; digital; dysbiosis; efficacy testing; experimental study; gastrointestinal; grape seed; histological image; histological specimens; imaging system; improved; in vivo; instrument; live cell imaging; luminescence; lung cancer cell; lung tumorigenesis; medical schools; metabolic abnormality assessment; microscopic imaging; mouse model; non-opioid analgesic; opioid overdose; opioid use disorder; pain model; painful neuropathy; preclinical study; professor; protein expression; response; salt intake; sex; skin disorder; symptomatology; therapeutic target; treatment response; user-friendly; wasting

Many veterans are burdened with chronic and complex medical conditions ranging from post-traumatic stress disorder, chronic pain, cancer, obesity, pre-diabetes mellitus, frank diabetes, acute kidney injury, chronic kidney disease, end-stage renal disease, to a host of sclerotic disorders. This application is for a Cytation 5 Cell Imaging Multi-Mode Reader Imaging System with connected BioSpa 8 Automated Incubator and Robotic Arm that will be used in experiments addressing the following Aims of Major Users toward combating these health issues: 1. Karin Westlund High, Ph.D. “Modulating Pain Generators of Chronic Trigeminal Neuropathic Pain”25% Test efficacy of the extended release NAα1R antagonist doxazosin for reduction of chronic pain/anxiety/ depression related behaviors. Chronic pain has poor therapeutic response to analgesics, especially opioids, and remains a significant challenge. Veteran patients report more severe chronic pain than non-veteran patients and are twice as likely to die of an opioid overdose in attempts to find relief. There is an urgent need to identify chronic pain induced brain RNA/protein expression signatures to assist in the development of non-opioid pharmaceutical compounds. The proposed studies are to determine efficacy of the extended release NAα1R antagonist doxazosin when tested in weeks 8-10. Automated imaging of histological samples with the requested imaging system will assess cellular and biochemical changes along the pain neuraxis and significantly facilitate experimental progress. 2. Jenny Mao, M.D. “Combinations of Grape Seed and Milk Thistle Extracts Against Lung Cancer” 25% Determine the maximum tolerated dose (MTD), PK/PD, and anti-cancer effects of leucoselect phytosome (LP) with siliphos in murine models. Historically, the VA has emphasized oncology research, particularly finding new treatments. Goal is to systematically determine the synergistic, antineoplastic properties of combinations of a standardized grape seed procyanidin extract (GSE), leucoselect phytosome (LP), and milk thistle silymarin extract (MTE), siliphos, against lung cancer using both in vitro and in vivo preclinical models. We hypothesize that oral administration of LP and siliphos will synergistically inhibit cancer growth and induce apoptosis in human lung cancer xenograft models, via favorable modulations of mechanisms associated with lung tumorigenesis/promotion. Effects of varying doses on proliferation and apoptosis in co-cultured cells over time will be determined and correlated to levels of GSE, MTE, and metabolites. The Cytation 5 Imaging System will enable automated repeated imaging of cultured cells with different treatments for PK/PD studies. 3. Manoocher Soleimani, M.D. “The Role of Collecting Duct Chloride Transporters in Salt Absorption and Blood Pressure Momeostasis” 25%. Ascertain the role of chloride transporter KBAT in salt absorption in the collecting duct in pathophysiologic states. Many veterans are admitted to the hospital with altered systemic blood pressure or excess fluid. The kidney’s distal nephron of the kidney plays a central role in vascular volume homeostasis and blood pressure regulation by regulating the absorption of salt via specialized molecules. We hypothesize that kidney brain anion transporter (KBAT) plays an important role in salt absorption in the entire collecting duct following increased delivery of salt consequent to the inactivation/inhibition of salt absorbing transporter proteins or in response to increased dietary salt intake. Using a combination of genetically engineered mouse models, metabolic studies, blood pressure recording, and molecular studies, we will determine the role of KBAT in salt absorption in pathophysiologic states. The requested imaging system will accelerate acquisition and analysis of transporter protein expression in kidney sections under different experimental conditions. Projects of Minor Users will monitor mitophagy through colocalization of mitochondrial probes with autophagy biomarkers, quantify cellular and molecular mechanisms that lead to gastrointestinal dysbiosis, hospital-acquired fungal infections, chronic kidney disease and opioid use disorder using the Cytation 5 Imaging System.

许多退伍军人都背负着慢性和复杂的医疗条件，从创伤后压力， 疾病、慢性疼痛、癌症、肥胖、糖尿病前期、明确的糖尿病、急性肾损伤、慢性肾损伤 疾病，终末期肾病，到一系列的神经系统疾病。此应用程序适用于Cytation 5细胞成像 连接BioSpa 8自动化培养箱和机械臂的多模式读取器成像系统， 用于解决主要用户对抗这些健康问题的以下目标的实验： 1. Karin Westlund高中博士“调节慢性三叉神经痛的疼痛发生器“25% 缓释NAα 1 R拮抗剂多沙唑嗪减轻慢性疼痛/焦虑/抑郁的疗效试验 抑郁症相关行为慢性疼痛对镇痛药，特别是阿片类药物的治疗反应较差， 并且仍然是一个重大的挑战。退伍军人患者比非退伍军人患者报告更严重的慢性疼痛 并且在试图寻求缓解时死于阿片类药物过量的可能性是其他人的两倍。迫切需要查明 慢性疼痛诱导的脑RNA/蛋白质表达特征有助于非阿片样物质的发展 药物化合物。拟定研究旨在确定缓释NAα 1 R的疗效 拮抗剂多沙唑嗪时，测试在第8-10周。组织学样本的自动成像， 成像系统将评估沿着疼痛神经轴的细胞和生化变化， 实验进展。 2. Jenny Mao，医学博士“葡萄籽和牛奶蓟提取物的组合对肺癌”25% 确定leucoselect phytosome的最大耐受剂量（MTD）、PK/PD和抗癌作用 (LP)在小鼠模型中进行了实验。从历史上看，VA一直强调肿瘤学研究，特别是 寻找新的治疗方法目的是系统地确定协同，协同性能， 标准化葡萄籽原花青素提取物（GSE）、白细胞选择植物体（LP）和牛奶的组合 蓟水飞蓟素提取物（MTE），siliphos，使用体外和体内临床前模型对抗肺癌。 我们假设LP和硅磷的口服给药将协同抑制癌症生长并诱导肿瘤细胞凋亡。 人肺癌异种移植模型中的细胞凋亡，通过与 肺肿瘤发生/促进。不同剂量对体外共培养细胞增殖和凋亡的影响 将确定时间并将其与GSE、MTE和代谢物的水平相关联。Cytation 5成像系统 将使培养细胞的自动重复成像与不同的治疗PK/PD研究。 3. Manoocher Soleimani，医学博士“收集管氯化物转运蛋白在盐吸收和 血压稳定”25%。确定氯离子转运蛋白KBAT在盐吸收中的作用 病理生理状态下的集合管。许多退伍军人被送进医院， 全身血压或过量液体。肾脏的远端肾单位在血管生成中起着中心作用。 通过特殊分子调节盐的吸收来调节容量稳态和血压调节。 我们假设肾脑阴离子转运蛋白（KBAT）在整个脑组织的盐吸收中起着重要作用。 由于盐吸收的失活/抑制而增加的盐输送后的集合管 转运蛋白或响应增加的膳食盐摄入。利用基因工程和 小鼠模型、代谢研究、血压记录和分子研究，我们将确定 KBAT在盐吸收中的作用。所要求的成像系统将加速采集 以及在不同实验条件下肾切片中转运蛋白表达的分析。 小用户项目将通过线粒体探针与自噬的共定位来监测线粒体自噬 生物标志物，量化导致胃肠道生态失调的细胞和分子机制， 真菌感染、慢性肾脏疾病和阿片类药物使用障碍。