RED CELL PRESERVATION

红细胞保存

• 批准号: 2221739
• 负责人: TIBOR J GREENWALT
• 金额: $ 12.73万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2221739
• 关键词: SDS polyacrylamide gel electrophoresis; acidity /alkalinity; adenine nucleotides; adenosine triphosphate; ammonium chloride; blood banks; blood donor; blood lipoprotein metabolism; blood preservation; carbohydrate metabolism; cell age; diphosphoglycerate; erythrocyte membrane; erythrocytes; glycolysis; hemolysis; human subject; oxidation; phosphates; phosphatidylinositols; phospholipids; radiotracer; solutions; thin layer chromatography

The long term goal is to extend the useful shelf life of human red blood cells. The specific aims are: l) to study the effect of ammonium (NH4+) and phosphate (Pi) on nucleotide and carbohydrate metabolism of RBCs; 2) to study membrane vesiculation and aminophospholipid flip flop during blood bank storage; 3) to identify correlated changes in the phosphinositides; 4) to ascertain the development of protein oxidation during storage; 5) to perform autologous in vivo viability studies. Units of packed red blood cells (PRBCs) will be stored with experimental additive solutions (EASs) in which the concentrations of NH4+ and Pi will be varied. The in vitro studies will include measurements at intervals up to 12 weeks of adenosine triphosphate 2,3-diphosphoglycerate, mean corpuscular volume, hemolysis, vesiculation and pH. 51Cr-labeled RBCs will be used in paired viability studies comparing selected EASs with a commercial additive solution, Adsol. For the metabolic studies, adenine- 8-14c will be used to measure the turnover of purine nucleotides and nucleosides. The levels of glucose-6-PO4 and fructose-6-PO4 will be used as indicators of phosphofructokinase activity. The ratios of lactate to pyruvate and the oxidized to the reduced form of nicotinamide adenine dinucleotide (NAD:NADH) will be used to gauge the activity of 3- phosphoglyceraldehyde dehydrogenase. Trinitrobenzenesulfonic acid will be used to quantitate the change in accessibility of phosphatidyl serine and ethanolamine during storage. Thin layer chromatography will be performed to separate and quantify the phosphinositides. Spectrin oxidation will be assessed using thiol disulfide exchange chromatography followed by gel electrophoresis. These studies will serve to enhance understanding of the metabolic changes affecting RBCs during blood bank storage. The resulting health benefits will be: 1) more effective operation of autologous donor programs. Longer effective shelf life of RBCs will make it possible to store more units preoperatively if needed and permit longer intervals between donations when the patient's clinical status makes this desirable; 2) the longer shelf life will make it possible to maintain the blood inventories of outlying hospitals more effectively with less outdating. Loss of blood by outdating still averages 10 percent in such situations.

长期目标是延长人类红血的有用货架 细胞。具体目的是：L）研究铵的作用（NH4+） RBC的核苷酸和碳水化合物代谢上的磷酸盐（PI）； 2） 研究膜囊泡和氨基磷脂触触 血库存储； 3）确定相关的变化 磷酸糖苷； 4）确定蛋白质氧化的发展 存储期间； 5）进行体内生存力研究。单位 填充的红细胞（PRBC）将使用实验存储 NH4+和PI浓度的添加溶液（EASS） 变化。体外研究将包括时间间隔 长达12周的三磷酸腺苷2,3-二磷酸，平均值 角球体积，溶血，囊泡和pH。 51 cr标记的RBC 将用于比较选定的EAS与A的配对生存力研究 商业添加剂解决方案，Adsol。对于代谢研究，腺嘌呤 - 8-14C将用于测量嘌呤核苷酸和 核苷。将使用葡萄糖-6-PO4和果糖-6 -PO4的水平 作为磷酸果糖激酶活性的指标。乳酸的比率 丙酮酸和氧化为烟酰胺腺嘌呤的还原形式 二核苷酸（NAD：NADH）将用于评估3-的活性 磷酸甘油醛脱氢酶。三硝基磺酸会 用于定量磷脂酰丝氨酸的可及性的变化 和储存过程中的乙醇胺。薄层色谱法将是 进行分离并量化磷酸糖苷。光谱 氧化将使用硫醇二硫化物色谱进行评估 然后是凝胶电泳。这些研究将有助于增强 了解影响血库中RBC的代谢变化 贮存。由此产生的健康益处将是：1）更有效 自动捐赠计划的操作。更长的有效货架寿命 RBC可以在需要时在术前存储更多单位 并允许在患者临床时捐赠之间更长的间隔 状态使这一理想； 2）较长的保质期会成功 可以维持更多的医院血液清单 有效的过时。静止的血液流失 在这种情况下，平均为10％。