Analytical Performance/Clinical Utility Of Laboratory Tests For Atherothrombosis

动脉粥样硬化血栓形成实验室测试的分析性能/临床实用性

• 批准号: 7593108
• 负责人: Gyorgy Csako
• 金额: $ 2.6万
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7593108
• 关键词: Bacterial Adhesion; Blood; Brain natriuretic peptide; C-reactive protein; Cardiac; Cardiovascular system; Cell Count; Cell Survival; Cells; Centrifugation; Chemistry; Cholesterol; Chorionic Gonadotropin, beta Subunit; Clinical; Collection; Coronary Arteriosclerosis; Corticotropin; Creatine; Defense Mechanisms; Diagnosis; Edetic Acid; Electrophoresis; Excision; Failure; Fibrinogen; Fibronectins; Freezing; Gel; Goals; Hemostatic Agents; Heparin; High Density Lipoprotein Cholesterol; Human; Human Chorionic Gonadotropin; Hydrocortisone; Hypertriglyceridemia; Immunoassay; Infection; Inflammatory; Isoenzymes; LDL Cholesterol Lipoproteins; Laboratories; Lipids; Lipoproteins; Major Depressive Disorder; Mammalian Cell; Mediating; Methods; Minority; Monitor; Mus; Nature; Nitric Oxide; Patients; Performance; Plasma; Play; Population; Potassium; Preparation; Process; Proteins; Protocols documentation; RNA Splicing; Rehabilitation therapy; Rheumatoid Arthritis; Risk; Risk Factors; Rodent; Role; Sampling; Sepsis; Serum; Serum amyloid A protein; Source; Specimen; Speed; Stem cells; Temperature; Testing; Therapeutic Intervention; Thyrotropin; Triglycerides; Troponin I; Troponin T; Tube; Variant; Woman; apolipoprotein Lp(a+); atherothrombosis; cell type; cold temperature; cryogel; cysteine rich protein; disorder risk; high density lipoprotein receptor; lipoprotein cholesterol; lipoprotein triglyceride; receptor; response; scavenger receptor

Atherothrombosis is a multifactorial disease and risk factors for atherothrombosis are now estimated to be in the hundreds. In addition to classical risk factors like serum lipids (total cholesterol and triglycerides), lipoproteins, and apolipoproteins, a variety of inflammatory molecules such as C-reactive protein and serum amyloid A, hemostatic factors and various specific cell types have been also considered for risk and/or require management. Studying the cardiovascular lipid profile of patients with rheumatoid arthritis (RA), we observed diagnostically important laboratory interferences and studied their nature and source. Initially, we attempted to quantitate lipoprotein-cholesterol fractions in previously frozen heparinized plasma of these patients using an electrophoretic enzymatic cholesterol method (REP Vis Cholesterol, Helena). However, our attempt has failed because of incomplete separation of lipoproteins in all 54 specimens tested, apparently due to the phenomenon of cryogelation. Cryogel is a co-precipitate of extra domain A-containing (cellular) fibronectin, plasma fibronectin, fibrinogen and heparin. Cryogelation has been described to occur in the presence of heparin at low temperatures and has been found to be common in patients with RA. Our failure with the electrophoretic lipoprotein-cholesterol test prompted studying possible interference of cryogelation with other laboratory tests. Serum and heparinized plasma were prepared from 6 RA patients using different blood collection and blood processing/storage protocols. These protocols included preparation of serum from separator gel tube and preparation of heparinized and EDTA plasma at room temperature and at 37 oC and storage of the specimens in refrigerator or frozen at -70 oC prior to laboratory testing. A panel of 20 routine chemistry tests and tests for total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were performed using the Synchron LX-20 automated chemistry analyzer (Beckman Coulter). Select sera and plasmas were also analyzed for troponin I, thyroid-stimulating hormone (TSH), adrenocorticotropic hormone or ACTH and cortisol (Immulite 2500, DPC/Siemens), troponin T, pro-brain natriuretic peptide (BNP), creatine kine-MB or CK-MB isoenzyme, and beta-human chorionic gonadotropin or beta-HCG (Elecsys 2010, Beckman Coulter), brain natriuretic peptide or BNP (Axsym, Abbott), high-sensitivity C-reactive protein or hsCRP and rheumotaid factor or RF (Immage, Beckman Coulter), serum amyloid A or SAA (BN-II, Dade-Behring), and electrophoretic protein fractions (Hydragel, Sebia). The presence of visible cryogel in heparinized plasma specimens from the heparinized plasma specimens necessitated high-speed centrifugation prior to testing. Apart from expected differences between serum and plasma for potassium and total protein, the results for most other routine chemistry tests, TC, TG, various immunoassays and protein electrophoresis were insignificantly different, either statistically or clinically, among various protocols. Compared to the reference serum specimens (serum processed and stored at 4 oC), all refrigerated or frozen heparinized plasma specimens gave higher (8%) HDL-C results and tended to give lower (4%) LDL-C results; with more pronounced effects for heparinized plasma specimens stored and processed at 37oC. It appears that, apart from lipoprotein fraction testing, even refrigerated or previously frozen heparinized plasma samples are suitable for most routine and specialized chemistry tests after removal of the cryogel that may cause sampling problems. In a collaborative study, we found that cardiac rehabilitation in patients receiving stable statin therapy and with low-density lipoprotein cholesterol at goal increases endothelial progenitor cell number, endothelial progenitor cell survival, and endothelial differentiation potential, associated with increased nitric oxide in the blood. Although this response was observed in most patients, a significant minority showed neither endothelial progenitor cell mobilization nor increased nitric oxide in the blood. In another collaborative study, we found that both serum SAA and CRP are significantly elevated in unmedicated, remitted women with major depressive disorder (MDD) versus controls. These findings suggested a sustained low-grade pro-inflammatory state in women with MDD that may be related to the increased coronary artery disease (CAD) risk observed previously in this population. In still another collaborative study, we studied the ability of the rodent (mouse) and human scavenger receptors SR-BI (called CLA-1 in humans) and its splicing variant SRB-II (called CLA-2 in humans) to mediate bacterial adhesion and cytosolic bacterial invasion in mammalian cells. Our results indicate that these receptors, in addition to their already established function as high-density lipoprotein receptors, also are involved in the defense mechanism of mammalian host cells and, by facilitating bacterial adhesion and cytosolic invasion, they may play an important role in infection and sepsis.

动脉粥样硬化血栓形成是一种多因素疾病，目前估计动脉粥样硬化血栓形成的危险因素有数百种。除了经典的风险因素如血清脂质（总胆固醇和甘油三酯）、脂蛋白和载脂蛋白外，各种炎症分子如C-反应蛋白和血清淀粉样蛋白A、止血因子和各种特定细胞类型也被认为是风险和/或需要管理。研究类风湿性关节炎（RA）患者的心血管血脂谱，我们观察到诊断重要的实验室干扰，并研究其性质和来源。最初，我们尝试使用电泳酶促胆固醇法（REP维斯胆固醇，Helena）定量这些患者先前冷冻的肝素化血浆中的脂蛋白-胆固醇组分。然而，我们的尝试失败了，因为在所有54个样本中的脂蛋白分离不完全，显然是由于冷冻现象。Cryogel是含有（细胞）纤连蛋白、血浆纤连蛋白、纤维蛋白原和肝素的额外结构域A的共沉淀物。据描述，在肝素存在的情况下，在低温下会发生冷冻，并且在RA患者中很常见。我们在电泳脂蛋白胆固醇试验中的失败促使我们研究冷冻疗法对其他实验室检查的可能干扰。使用不同的血液采集和血液处理/储存方案从6名RA患者制备血清和肝素化血浆。这些方案包括从分离凝胶管制备血清，在室温和37 ℃下制备肝素化和EDTA血浆，并在实验室检测前将标本储存在冰箱中或在-70 ℃下冷冻。使用Synchron LX-20自动化学分析仪（Beckman Coulter）进行一组20项常规化学检查和总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白胆固醇（HDL-C）和低密度脂蛋白胆固醇（LDL-C）检查。选择血清和血浆进行肌钙蛋白I，促甲状腺激素（TSH），促肾上腺皮质激素或ACTH和皮质醇的分析（Immulite 2500，DPC/Siemens）、肌钙蛋白T、脑钠肽前体（BNP）、肌酸激酶-MB或CK-MB同工酶和β-人绒毛膜促性腺激素或β-HCG（Elecsys 2010，Beckman Coulter）、脑钠肽或BNP（Axsym，Abbott）、高敏C反应蛋白或hsCRP和类风湿因子或RF（Immage，Beckman Coulter）、血清淀粉样蛋白A或SAA（BN-II，Dade-Behring）和电泳蛋白级分（Hydragel，Sebia）。由于肝素化血浆标本中存在可见的冻凝胶，因此需要在检测前进行高速离心。除了钾和总蛋白的血清和血浆之间的预期差异外，大多数其他常规化学检查、TC、TG、各种免疫测定和蛋白电泳的结果在各种方案之间无统计学或临床差异。与参考血清标本（4 ℃下处理和储存的血清）相比，所有冷藏或冷冻肝素化血浆标本的HDL-C结果均较高（8%），LDL-C结果往往较低（4%）;在37 ℃下储存和处理的肝素化血浆标本的影响更明显。似乎除了脂蛋白部分检测外，即使是冷藏或先前冷冻的肝素化血浆样本，在去除可能导致采样问题的冷冻凝胶后，也适用于大多数常规和专业化学检测。 在一项合作研究中，我们发现接受稳定他汀类药物治疗并以低密度脂蛋白胆固醇为目标的患者的心脏康复增加了内皮祖细胞数量，内皮祖细胞存活率和内皮分化潜能，与血液中一氧化氮的增加有关。虽然在大多数患者中观察到这种反应，但少数患者既没有内皮祖细胞动员，也没有增加血液中的一氧化氮。在另一项合作研究中，我们发现与对照组相比，未服药、缓解的重度抑郁症（MDD）女性的血清SAA和CRP均显着升高。这些发现表明，MDD女性患者持续的低级别促炎状态可能与先前在该人群中观察到的冠状动脉疾病（CAD）风险增加有关。在另一项合作研究中，我们研究了啮齿动物（小鼠）和人类清道夫受体SR-BI（在人类中称为CLA-1）及其剪接变体SRB-II（在人类中称为CLA-2）介导哺乳动物细胞中细菌粘附和胞质细菌入侵的能力。我们的研究结果表明，这些受体，除了他们已经建立的高密度脂蛋白受体的功能，也参与了哺乳动物宿主细胞的防御机制，通过促进细菌粘附和胞质入侵，他们可能在感染和败血症中发挥重要作用。