## 3 Results
### Chemical Comparison and Mock Treatment Are Inaccurate Markers of Cellular Responses
Mock treatment reduced viability and stimulated mitochondrial O\({}_{2}\)\({}^{-}\) production above the untreated control only at H\({}_{2}\)O\({}_{2}\) concentrations above 100 \(\upmu\)M. (**d**) There were no changes in ecto-CRT responses to mock treatments. The mock treatment experiments were performed once in triplicate. The data represent the mean and standard deviation. Our results demonstrated that, for Jurkats, mock treatment was not a good indicator of the role plasma components have in triggering cellular responses. The H\({}_{2}\)O\({}_{2}\) concentrations produced by both devices should trigger stronger cellular responses. The reduced toxicity of H\({}_{2}\)O\({}_{2}\) during plasma treatment was likely due to secondary reactions in the medium itself. Components in the medium and the cells themselves that offer oxidizable targets for plasma-generated species (both short and long-lived) altered the concentrations of individual species. Several investigations focus on the reactions between individual amino acids and other organic compounds, such as reactive sulfur species with plasma-generated RONS [51; 52; 53]. These compounds reduce the amount of plasma-generated species in the liquid by reacting with them. However, subsequent reactions can lead to increases in the overall inventory of RONS in the solution. [PERSON] et al. showed that cysteine alone could reduce H\({}_{2}\)O\({}_{2}\) produced by the kINPen while increasing reactive nitrogen species (RNS) [52]. As [PERSON] et al. showed through electroparamagnetic spectroscopy, there are also increases in the concentrations of short-lived species after plasma treatment of cell culture medium [45], which are precursors to these long-lived species [44]. The biological effects of the short-lived species on cellular responses are well-documented in plasma literature [54; 35; 15]. These initiating molecules are absent during mock treatments, which is why we did not observe similar cellular responses and GSH modifications to plasma treatment. Without the short-lived species and the organic compounds they modify, our mock treatment with exogenous H\({}_{2}\)O\({}_{2}\) and NO\({}_{2}\)\({}^{-}\) is not representative of the actual contribution these long-lived species have during plasma treatment. GSH Modifications Unique to Plasma Source Indicate the Stressors and the Potential for Cellular Response Present During Plasma Treatment  
The data presented thus far indicated a lack of correlation between the relatively easy-to-measure long-lived species and cellular responses. The results from mock treatments demonstrated that using the measured RONS as an indicator of biological responses to plasma treatment was not sufficient. Therefore, there was a need for a method that is a true reflection of plasma parameters and the ensuing biological outcomes. In subsequent experiments, we expanded on the use of GSH as an indicator of how plasma might impact the cellular redox system, as proposed by [PERSON] et al. [23], and related our measured biological responses to measured GSH modifications. GSH modifications after plasma or mock treatment were analyzed to address their correlation with the observed biological responses. The specific modifications to GSH exposed to plasma treatment elucidated the possible reactive species near the molecule. Similar to previous reports on microsecond pulsed DBD-treated GSH [23], nspDBD plasma treatment caused a decrease in the intensity of the peak at 2560 cm\({}^{-1}\) and an increase of the 1045 cm\({}^{-1}\) peak when compared to the untreated GSH (Figure 7). These wavenumbers corresponded to the S-H (v (S-H)) and the S=O (v (S=O)) vibrational stretching modes, respectively, and indicated that S-H bonds were oxidized. Molecular dynamic (MD) simulations have previously shown that H\({}_{2}\)O\({}_{2}\) did not initialize the oxidation of glutathione [23].