Metabolic Profiling of a Porcine Combat Trauma-Injury Model Using NMR and Multi-Mode LC-MS Metabolomics-A Preliminary Study.

Profiles of combat injuries worldwide have shown that penetrating trauma is one of the most common injuries sustained during battle. This is usually accompanied by severe bleeding or hemorrhage. If the soldier does not bleed to death, he may eventually succumb to complications arising from trauma hemorrhagic shock (THS). THS occurs when there is a deficiency of oxygen reaching the organs due to excessive blood loss. It can trigger massive metabolic derangements and an overwhelming inflammatory response, which can subsequently lead to the failure of organs and possibly death. A better understanding of the acute metabolic changes occurring after THS can help in the development of interventional strategies, as well as lead to the identification of potential biomarkers for rapid diagnosis of hemorrhagic shock and organ failure. In this preliminary study, a metabolomic approach using the complementary platforms of nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography coupled with mass spectrometry (LC-MS) was used to determine the metabolic changes occurring in a porcine model of combat trauma injury comprising of penetrating trauma to a limb with hemorrhagic shock. Several metabolites associated with the acute-phase reaction, inflammation, energy depletion, oxidative stress, and possible renal dysfunction were identified to be significantly changed after a thirty-minute shock period.

Rapid detection of botulinum neurotoxins A, B, E, and F by optical immunoassay.

Botulism is caused by the neurotoxins (BoNTs) produced from Clostridium botulinum. These neurotxins often lead to fatal neuroparalytic disease which is regarded a major threat to the public health. For this reason, rapid and reliable diagnosis of BoNTs in field settings and peripheral care centers is highly valuable. Here, we describe a multiplexed and sensitive optical immunoassay (OIA) for the rapid detection of four medically important BoNTs (A, B, E, and F). The assay is based on detection of physical changes in the thickness of molecular thin film resulting from specific immunobinding events on an optical silicon chip. The immunocomplex causes destructive interference of a particular wavelength of reflected white light from gold to purple-blue on an optical surface depending on the concentration of the analyte. This test allows simultaneous characterization of toxin type and semi-quantitative assessment of toxin level. The assay was four times more sensitive than ELISA when performed with the same reagents. The limit of detection (LOD) for the BoNTs was, respectively, 2.5-5 ng/mL, 5-10 ng/mL, and 10-20 ng/mL in experimentally spiked buffer, water and food matrices. The less logistic load and visual read-out of this method promises potential applicability of this assay in the field as well as in a clinical settings.