Definición de variables

Here, we report the results of the first integrated study using differentiated HL-60 cells as a model system for investigating whether changes in ultra-weak photon emission are correlated with changes in the metabolic profile. Specifically, HL-60 cells were differentiated into neutrophil-like cells and induced to undergo respiratory burst. The temporal UPE profile was recorded during respiratory burst, and cationic metabolites were measured in cell extracts and used to create a metabolomics profile. Our results revealed that several metabolites are significantly correlated with changes in UPE. Although these findings suggest that the methionine pathway plays a role in UPE, additional experiments are needed in order to elucidate the precise role of methionine metabolism in UPE. Therefore, future

Tracking biochemical changes correlated with UPE using metabolomics

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studies should use analytical platforms that include additional metabolites, including metabolites that are associated with oxidative stress, oxylipins, and cellular energy levels, thereby providing a more comprehensive metabolic profile.

In summary, we provide the first experimental evidence that measuring UPE can be used to study metabolic profiles. Specifically, we found that the concentration profiles of putrescine, glutathione, sarcosine, creatine, and β-alanine were significantly correlated with the UPE signal, suggesting that the methionine pathway plays a key role in UPE. These results pave the way for future research regarding metabolic pathways that underlie UPE, and they provide important information regarding the potential use of UPE as a diagnostic tool. ACKNOWLEDGMENTS

We thank Ondřej Kučera and Cristiano de Mello Gallep for valuable comments and detailed feedback regarding the manuscript. R.C.R.B. was supported by the Brazilian Scholarship Program “Science without Borders” of the Brazilian National Council for Scientific and Technological Development (Conselho Nacional de Desenvolvimento Científico e Tecnológico, fellowship no. 230827/2012-8). K.Č. and M.C. were supported by the Czech Science Foundation (grant no. GP13-29294S).

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