Identification of novel CSF biomarkers for neurodegeneration and their validation by a high-throughput multiplexed targeted proteomic assay

BACKGROUND: Currently there are no effective treatments for many neurodegenerative diseases. Reliable biomarkers for identifying and stratifying these diseases will be important in the development of future novel therapies. Lewy Body Dementia (LBD) is considered an under diagnosed form of dementia for which markers are needed to discriminate LBD from other forms of dementia such as Alzheimer's Disease (AD). This work describes a Label-Free proteomic profiling analysis of cerebral spinal fluid (CSF) from non-neurodegenerative controls and patients with LBD. Using this technology we identified several potential novel markers for LBD. These were then combined with other biomarkers from previously published studies, to create a 10 min multiplexed targeted and translational MRM-LC-MS/MS assay. This test was used to validate our new assay in a larger cohort of samples including controls and the other neurodegenerative conditions of Alzheimer's and Parkinson's disease (PD). RESULTS: Thirty eight proteins showed significantly (p < 0.05) altered expression in LBD CSF by proteomic profiling. The targeted MRM-LC-MS/MS assay revealed 4 proteins that were specific for the identification of AD from LBD: ectonucleotide pyrophosphatase/phosphodiesterase 2 (p < 0.0001), lysosome-associated membrane protein 1 (p < 0.0001), pro-orexin (p < 0.0017) and transthyretin (p < 0.0001). Nineteen proteins were elevated significantly in both AD and LBD versus the control group of which 4 proteins are novel (malate dehydrogenase 1, serum amyloid A4, GM2-activator protein, and prosaposin). Protein-DJ1 was only elevated significantly in the PD group and not in either LBD or AD samples. Correlations with Alzheimer-associated amyloid β-42 levels, determined by ELISA, were observed for transthyretin, GM2 activator protein and IGF2 in the AD disease group (r(2) ≥ 0.39, p ≤ 0.012). Cystatin C, ubiquitin and osteopontin showed a strong significant linear relationship (r(2) ≥ 0.4, p ≤ 0.03) with phosphorylated-tau levels in all groups, whilst malate dehydrogenase and apolipoprotein E demonstrated a linear relationship with phosphorylated-tau and total-tau levels in only AD and LBD disease groups. CONCLUSIONS: Using proteomics we have identified several potential and novel markers of neurodegeneration and subsequently validated them using a rapid, multiplexed mass spectral test. This targeted proteomic platform can measure common markers of neurodegeneration that correlate with existing diagnostic makers as well as some that have potential to show changes between AD from LBD

The relationship between intracranial pressure and brain oxygenation following traumatic brain injury in sheep

BackgroundWhile it is understood that raised intracranial pressure (ICP) after traumatic brain injury (TBI) may negatively impact on brain tissue oxygenation (PbtO2), few studies have characterized the inter-relationship between these two variables, particularly in a large animal model that replicates the human gyrencephalic brain. The current study uses an ovine model to examine the dynamics of ICP and PbtO2 after TBI.Materials and methodsFive 2-year-old male Merino sheep were anesthetized with isoflurane and impacted in the left temporal region using a humane stunner. ICP and PbtO2 were then monitored over the following 4 h using a Codman ICP Express monitoring system and a LICOX brain tissue oxygen monitoring system, respectively. Two additional sheep were anesthetized and monitored as sham (uninjured) controls.FindingsMean ICP 60 min following TBI was over 25 mmHg (p ConclusionsOur results suggest that TBI results in early changes in ICP that are associated with profound declines in PbtO2, and may indicate the need for earlier management of ICP after TBI.Robert Vink, Kartik D. Bahtia, Peter L. Reill

Geochemical and Nd isotope compositions of detrital sediments on the north margin of the South China Sea: provenance and tectonic implications

A major re‐organization of regional drainages in eastern Tibet and south‐western China took place in the Cenozoic as deformation from the growing Himalayas and Tibetan Plateau affected an increasingly wider area. The effects of these changes on the regional sediment routing systems is not well constrained. This study examines the geochemical and Nd signatures of sedimentary rocks from the Ying‐Qiong and Nanxiong basins on the northern margin of the South China Sea to constrain and identify any significant changes in sediment source. Upper Cretaceous to Lower Eocene sedimentary rocks in the Nanxiong Basin show higher Th/Sc, La/Sc, Th/Cr and Th/Co ratios and lower Eu/Eu* ratios than PAAS (post‐Archaean Australian Shale), which indicates that Palaeozoic sedimentary rocks of the South China Block were the main basin sediment source. In contrast, Oligocene to Pleistocene sedimentary rocks of the Ying‐Qiong Basin show an abrupt change in these trace‐element ratios between 16·3 and 10·4 Ma, indicating a mid‐Miocene shift in provenance. ɛNd values from the Ying‐Qiong Basin (range = −11·1 to −2·1) reinforce this, with pre‐13·8 Ma sedimentary rocks having average ɛNd of −5·6 (range = −2·1 to −7·4), and post‐13·8 Ma sedimentary rocks having average ɛNd of −9·3 (range = −8·7 to −11·1). During the Oligocene, the centre of rifting transferred south and basins on the north margin of the South China Sea experienced rapid subsidence. Further uplift and erosion then exposed Mesozoic and Cenozoic granites that supplied large amounts of granitic detritus, especially to the Ying‐Qiong Basin. Then a change occurred at ca 13 Ma resulting in less input from local sources (i.e. the fault blocks formed by Mesozoic‐Cenozoic tectonics and magmatism) to an increasing contribution of older continental material, mostly from Indochina to the west of the South China Sea

Photochemical generation of radicals from alkyl electrophiles using a nucleophilic organic catalyst

Chemists extensively use free radical reactivity for applications in organic synthesis, materials science, and life science. Traditionally, generating radicals requires strategies that exploit the bond dissociation energy or the redox properties of the precursors. Here, we disclose a photochemical catalytic approach that harnesses different physical properties of the substrate to form carbon radicals. We use a nucleophilic dithiocarbamate anion catalyst, adorned with a well-tailored chromophoric unit, to activate alkyl electrophiles via an S N 2 pathway. The resulting photon-absorbing intermediate affords radicals upon homolytic cleavage induced by visible light. This catalytic S N 2-based strategy, which exploits a fundamental mechanistic process of ionic chemistry, grants access to open-shell intermediates from a variety of substrates that would be incompatible with or inert to classical radical-generating strategies. We also describe how the method’s mild reaction conditions and high functional group tolerance could be advantageous for developing C–C bond-forming reactions, for streamlining the preparation of a marketed drug, for the late-stage elaboration of biorelevant compounds and for enantioselective radical catalysis