## 4 Discussion
\begin{table}
\begin{tabular}{c c c c c c c} \hline  & & \begin{tabular}{c} **Wet bDLE** \\ **(0.4 mg/mL)** \\ \end{tabular} & \begin{tabular}{c} **Wet bDLE** \\ **(0.4 mg/mL)** +** \\ **LPS (2 mg/mL)** \\ \end{tabular} & \begin{tabular}{c} **Dry bDLE** \\ **(0.4 mg/mL)** \\ \end{tabular} & \begin{tabular}{c} **Dry bDLE** \\ **(0.4 mg/mL)** \\ \end{tabular} & \begin{tabular}{c} **Dry bDLE** \\ **(0.4 mg/mL)** \\ \end{tabular} & \begin{tabular}{c} **Dry bDLE** \\ **(0.4 mg/mL)** \\ \end{tabular} & \
\begin{tabular}{c} **Dry bDLE** \\ **(0.4 mg/mL)** \\ \end{tabular} \\ \hline IL-12p70 (pg/mL) & 20.7 & 89.5 * & 20.7 & 28 & 29 & 29.1 \\ TNF-\(\alpha\) (pg/mL) & 49.5 & 8530 * & 1078 & 7.8 & 1050 & 0 \\ IFN-\(\gamma\) (pg/mL) & 4.8 & 2.9 & 3 & 3 & 3 & 3 \\ MCP-1 (pg/mL) & 70.2 & 1089 * & 103.2 & 77.4 & 89.6 & 90.1 \\ IL-10 (pg/mL) & 50.5 & 75.9 & 55.9 & 67.3 & 67.3 & 388.1 \\ IL-6 (pg/mL) & 21 & 14640 * & 1026 & 19.8 & 13.8 & 12 \\ \hline \end{tabular}
\begin{tabular}{c} Murine macrophages (\(5\times 10^{5}\)) were stimulated with LPS (2 mg/mL) and simultaneously treated with wet and dry bDLE. Cytokine levels were determined in the cell culture supernatant with the cytometric bead array mouse inflammation kit by flow cytometry. Statistical significance was determined by one-way ANOVA (* \(p<0.05\)).  
\end{table} 
Table 5: Cytokine levels in LPS stimulated murine macrophages treated with wet and dry bDLE. structural determination of the molecules present in both extracts, since these structures are determined by the intramolecular and intermolecular hydrogen bonds. The zeta potential values were slightly higher for wet bDLE; however, the values were positive at pH 4, 6, 7 and 10 for both extracts, which indicates high stability in solution and that the particles are dispersed (there is repulsion between the charged molecules and the ions found in the solution) [18]. Once we determined that there are structural differences between the wet and dry bDLE, we proceeded to evaluate whether there were any differences in their biological properties. Both extracts exhibited a MTT and ABTS radical cation scavenging ability, which indicates that the antioxidant components have not been inactivated by the heating process, and therefore maintain their therapeutic potential for the prevention and treatment of disorders where the oxidative free radicals are involved. The anti-inflammatory activity was evidenced by the decrease in free radicals (NO and ROS) and modulation of cytokines in cells stimulated with LPS and wet or dry bDLE. Previous results have demonstrated that wet bDLE induces cytokine expression in murine macrophages [19] and PBMC [20] and, when challenged with LPS, has the capacity to modulate inflammatory cytokines and mediators, including TNF-\(\alpha\) through a NF-\(\kappa\)B dependent mechanism [20]. bDLE also has an antioxidant capacity that has been previously reported [20], which can affect the inflammation process, as can be seen when comparing TNF-\(\alpha\) levels of wet bDLE and wet bDLE + LPS, against LPS alone. Complementary to this, our results show that the dry bDLE contains higher amounts of proteins, and it has been reported that the bioactive peptides present in food from vegetable and animal origin, including bovine milk, possess antioxidant and anti-inflammatory properties [21]. Furthermore, it has been proven that the processing method can significantly alter the biological activity, and that the activity of some peptides can be increased by heat through denaturation, while others could completely lose their activities after heating [22]. The anti-inflammatory mechanism for wet bDLE is seen for occurs through the regulation of Nrf and NF-\(\kappa\)B transcription factors [4; 19]. It remains to be elucidated whether dry bDLE shares the same action mechanism. We found selective cytotoxic activity against cancer cells for both wet and dry bDLE.