Fundamental Development of In-vivo NMR Technology to Understand Environmental Stress

目前的环境政策主要基于死亡、移动和生殖等最重要的终点，但很少提供关于亚致死影响、毒性机制或协同效应的信息。暴露于低水平污染物混合物是常态而不是例外，微妙的亚致死效应对动物、植物和人类群体可能更危险，因为它们往往只有在身体症状广泛传播后才被发现。医学领域最近确定环境暴露是许多神经退行性疾病的主要原因，包括与重金属和农药的联系，但确切的环境原因尚不清楚。 传统的自上而下的方法涉及识别所有污染物及其降解产物，然后在单个化合物的基础上评估毒性，鉴于我们环境的复杂性，这种方法极具挑战性。相反，自下而上的方法侧重于生物体本身，并提出以下问题：种群是否面临压力？;造成压力的压力源是什么？哪些生化途径会受到影响核磁共振（NMR）是现代研究中最强大的工具之一，可以在体内应用。因此，活生物体成为实时响应其环境的“终极生物传感器”，而NMR光谱仪解释生化变化，提供在分子水平上解释亚致死毒性的信息。最终目标是了解和分类复杂介质（例如工业废水，废水）和自然水生环境中的多方面应激反应，以便有一天，这些分子指纹可用于识别天然生物体中环境应激的确切原因。反过来，了解影响环境和人类健康的最成问题的压力因素，对于改善监测、有针对性的补救、预防和政策至关重要。 水生新生动物和卵特别容易受到毒素的影响，如果受到损害，整个种群和生态系统都可能受到影响。然而，由于其低灵敏度，目前的NMR方法需要100个新生儿/卵，使得常规筛查不切实际。 在这个提议中，微线圈NMR，微线圈阵列，新的有针对性的实验和应用将扩展到单个水生新生儿和鸡蛋在体内的NMR。由于信号增强和多重多个样品的能力，实验时间可以从数月缩短到小时/分钟。据我们所知，没有其他技术接近于提供如此丰富的体内分子信息。这一点，沿着科学问题变得越来越复杂，生物相关性始终处于最前沿的事实，很明显，这里开发的高分辨率体内NMR方法将成为下一代科学研究的关键。

Present environmental policies are based largely on apical endpoints such as death, movement and reproduction, but provide little information on sub-lethal impacts, toxic mechanisms or synergistic effects. Exposure to low-level contaminant mixtures is the norm rather than the exception, and subtle sub-lethal effects, are arguably more hazardous, to animal, plant and human populations, as they are often only detected, after physical symptoms become widespread. The medical field have recently identified environmental exposure as the predominant cause of many neurodegenerative diseases including links to heavy metals and pesticides, but the exact environmental causes are not yet known. A traditional top down approach, which involves identifying all contaminants, their degradation products, and then assessing toxicities on an individual compound basis is extremely challenging given the complexity of our environment. Conversely, a bottom-up approach focuses on the organisms themselves and asks questions such as; is a population stressed?; what are the stressor/stressors causing the stress?; and which biochemical pathways are impacted? Nuclear Magnetic Resonance (NMR) is one of the most powerful tools in modern research and can be applied in-vivo. As such the living organism becomes the “ultimate biosensor” responding in real-time to its environment, while the NMR spectrometer interprets the biochemical changes, providing information explaining sub-lethal toxicity at the molecular-level. The ultimate goal is to understand and categorize multifaceted stress responses in complex media (e.g. industrial effluent, waste water) and the natural aquatic environment, such that, one day, these molecular fingerprints can be used to identify the exact cause of environmental stress in native organisms. In turn, the knowledge as to the most problematic stressors impacting environmental and human health is essential to improving monitoring, targeted remediation, prevention and policies. Aquatic neonates and eggs are especially susceptible to toxins, and if compromised, entire populations and ecosystems can be impacted. However, due to its low sensitivity, current NMR approaches require 100's of neonates/eggs, making routine screening impractical. In this proposal, micro-coil NMR, micro-coil arrays, novel targeted experiments and applications will be developed to extend in-vivo NMR to single aquatic neonates and eggs. Due to the increased signal and ability to multiplex many samples, the experiment time can be reduced from months down to hrs/mins. To our knowledge no other techniques come close to providing such a wealth of molecular information in-vivo. This, along with the fact that scientific questions are becoming increasingly complex and biological relevance is always at the forefront, it is clear the high resolution in-vivo NMR methods developed here, will be key to the next generation of scientific research in general.