Metabolomics is a rapidly advancing field, and much of our understanding of the subject has come from research on cell lines. However, the results and interpretation of such studies depend on appropriate normalization of the data; ineffective or poorly chosen normalization methods can lead to frankly erroneous conclusions. That is a recurrent challenge because robust, reliable methods for normalization of data from cells have not been established. In this study, we have compared several methods for normalization of metabolomic data from cell extracts. Total protein concentration, cell count, and DNA concentration exhibited strong linear correlations with seeded cell number, but DNA concentration was found to be the most generally useful method for the following reasons: 1) DNA concentration showed the greatest consistency across a range of cell numbers; 2) DNA concentration was the closest to proportional with cell number; 3) DNA samples could be collected from the same dish as the metabolites; and 4) cell lines that grew in clumps were difficult to count accurately. We therefore conclude that DNA concentration is a widely applicable method for normalizing metabolomic data from adherent cell lines.

Objectives To evaluate student impressions of learning anatomy with mixed-reality and compare long-term information retention of female breast anatomy between students who learned with a mixed-reality supplement and their classmates who dissected cadavers. Methods In Part 1, 38 first-year medical student volunteers, randomly divided into two groups, completed a mixed-reality module and cadaveric dissection on the female breast in a counterbalanced design. Participants also completed post-quizzes and surveys. Part 2 was a non-randomized controlled trial, 8-months after completing Part 1 and 6-months after a final exam on this content. The performance of twenty-two Part 1 participants and 129 of their classmates, who only dissected, was compared on a delayed post-quiz. Wilcoxon signed-rank test, Mann-Whitney U test, and 95% confidence intervals were used to analyze the data. Results In Part 1, the Wilcoxon signed-rank test determined that participants expressed significantly more positive responses to mixed-reality and found mixed-reality easier for learning and teamwork. In Part 2, the Mann-Whitney U test found mixed-reality participants scored significantly higher on a delayed-post quiz than their classmates who only dissected (U = 928, p < .009). Conclusions This study suggests that medical students may prefer mixed-reality and that it may be an effective modality for learning breast anatomy while facilitating teamwork. Results also suggest that supplementing cadaveric dissection with mixed-reality may improve long-term retention for at least one anatomical topic. It is recommended that similar studies evaluate a larger sample and additional anatomical regions to determine the generalizability of these findings.

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