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We have recently observed that one-year use of high fat diet (HFD) that induced mild ketonemia lead to better learning and memory, larger hippocampi volumes without any changes to cortical volumes, as well as higher concentrations of total NAA (tNAA: N-acetylaspartate and N-acetylaspartateglutame; marker of neuronal viability), total Cho (tCho: Glycerophosphocholine +Phosphocholine, which are believed to be primarily involved in cell membrane breakdown and synthesis) and total Cr (tCr: creatine + phospo-creatine – involved in cell bioenergetics). However, the spectroscopic results may have been driven by specific processing procedures used by LC Model, thus we needed to use a different software to assure the obtained results are independent of processing procedure. TARQUIN (Wilson et al, 2011) [1] is an open source alternative that was demonstrated to work comparably well to LCModelTM with wide range of 1.5T and 3.0T proton spectra. However, it has not been used to process proton, animal spectra acquired at 7.0T. Here, we 1) created basis sets for TARQUIN to work with spectra obtained at 7T, 2) reanalyzed the data, and finally 3) compared performance of TARQUIN and LCModelTM for single voxel hippocampal and anterior cingulate cortex spectra obtained from 50 onemonth Wistar rats at 7T, and later, when they were one year old (n=47). Two different basis sets were proposed: one based on basis set used by LCModelTM, and the other one adapted from a basis set established for 16.1T. Given the intrinsic differences in processing between LCModelTM and TARQUIN, we evaluated quality of fit (Q) and performed Bland-Altman analysis to estimate the agreement between the methods. Moreover, we calculated mean baseline and mean fit for 50 one-month old rats to identify potential systematic errors in fits. Finally, results from an exemplary experiment obtained with LCModelTM were reproduced with TARQUIN. Bland Altman plots indicate that there is an acceptable agreement between LCModelTM and Tarquin with adjacent basis set for total N-Acetylo-aspartate (tNAA), total-choline (tCho), total-creatine (tCr) and glutamine/glutamate (Glx) (95% confidence interval of agreement below 20%). However, for both basis sets, Tarquin gave significantly more variable results in myo-Inositol comparing to LCModel. In conclusion, despite some potential biases to the results, spectra were successfully processed with Tarquin and they yielded similar results to those obtained with LCModel. Correspondence to: Bartosz Kossowski, Nencki Institute of Experimental Biology, ul. Pasteura 6, 02-661 Warsaw, Poland, Email: bartosz.kossowski@ gmail.com Tel: +48-22-6852-633

biology, engineering and medicine

Follow-up analyses on the effects of long-term use of high fat diet on hippocampal metabolite concentrations in Wistar rats: Comparing Tarquin quantification of 7.0T rat metabolites to LCModel