Medical treatment using high-voltage electric potential (HELP) devices to generate an electric field (EF) is an alternative therapy commonly used in Japan. However, the underlying mechanisms of the potential health benefits are not fully understood. To address this issue, we performed plasma lipidomics using liquid chromatography in combination with tandem mass spectrometry (LC-MS/MS). 9-Hydroxyoctadecadienoic acid (HODE), 13-HODE, and 13-hydroperoxy-octadecadienoic acid (HpODE) levels were significantly upregulated after HELP (18 kV, 30 min) exposure. However, there was no effect on HODE-related diol-metabolites, epoxidemetabolites, ketone-metabolites, or prostaglandins (PGs). We further examined the effect of HELP exposure on plasma concentrations of mediators using enzymelinked immunosorbent assay (ELISA)/enzyme immunoassay (EIA). Immunoreactive substance P (SP) and brain-derived neurotrophic factor (BDNF) levels were significantly upregulated after HELP exposure. Under these conditions, HELP exposure had no effect on immunoreactive levels of vasoactive intestinal peptide (VIP), bradykinin, calcitonin gene-related peptide (CGRP), or motilin. Our findings provide insight into the possible relationship between the pharmacological modulation of neuromediators and that of HODEs by EF exposure. They may also be important in the development of electroceuticals.

3-Hydroxybutyrate (beta-hydroxybutyrate or 3-HBA) is small lipid-derived molecule that serves as an energy carrier during fasting, and endogenous inhibitor of histone deacetylase (HDAC). Here we report the concentrations of 3-HBA in plasma samples obtained from healthy human subjects before and after exposure to a single 30-min high-voltage electric potential (HELP) exposure using quantitative capillary electrophoresis-time-of-flight mass spectrometry (CE-TOFMS). 2-and 3-HBA were significantly upregulated after HELP exposure (4.5 kV/electrode + 4.5 kV/electrode), but not after exposure to low-strength condition (2.25 kV/electrode + 2.25 kV/electrode). There was no effect on the concentrations of 2-hydroxybutanedioate, 2-hydroxytricarballylate, 2-oxopropanoate, or ATP. Because 3-HBA is known to induce histone acetylation at the forkhead box O3a (FoxO3a) promoter, we further confirmed that FOXO3A mRNA was upregulated by 3-HBA treatment in a human epithelial colorectal adenocarcinoma Caco-2 cell line. Under these conditions stated above, 3-HBA treatment had no effect on forkhead box P3 (FOXP3) or defensin beta 132 (DEFB132) mRNA expressions. Similar changes in FOXO3A mRNA expression were obtained using sodium 4-phenylbutyrate (4-PBA) or trichostatin A (TSA) instead of 3-HBA. Our findings provide a new insight into the epigenetic mechanisms of electric field (EF) therapy.

Medical treatment using high-voltage electric potential (HELP) devices to generate an electric field (EF) is an alternative therapy commonly used in Japan. However, the underlying mechanisms of the potential health benefits of this therapy are not fully understood. Therefore, we investigated the lysophosphatidylethanolamine (lysoPE) and lysophosphatidylinositol (lysoPI) levels using selected reaction monitoring (SRM) analysis in plasma samples obtained from healthy human subjects before and after exposure to a single HELP exposure (9 kV/electrode + 9 kV/electrode, 30 min). LysoPE-22:6 and lysoPE-20:4 were significantly upregulated after HELP exposure. However, there were no effects on the levels of phosphatidylethanolamine (PE) and phosphatidylinositol (PI), lysoPI, or other lysoPE species. LysoPE is known to activate G protein-coupled receptor 119 (GPR119). No X-ray crystal structure has been reported for GPR119; thus we examined the in silico docking simulation of lysoPE-22:6 with GPR119 using a homology model. LysoPE-22:6 showed strong interaction energy of -10.603 kcal/mol. Our findings provide new insight into the molecular mechanisms of the health benefits of EF therapy.

Although electric fields (EF) exert beneficial effects on animal wound healing, differentiation, cancers and rheumatoid arthritis, the molecular mechanisms of these effects have remained unclear about a half century. Therefore, we aimed to elucidate the molecular mechanisms underlying EF effects in Drosophila melanogaster as a genetic animal model. Here we show that the sleep quality of wild type (WT) flies was improved by exposure to a 50-Hz (35 kV/m) constant electric field during the day time, but not during the night time. The effect was undetectable in cryptochrome mutant (cryb) flies. Exposure to a 50-Hz electric field under low nutrient conditions elongated the lifespan of male and female WT flies by ~ 18%, but not of several cry mutants and cry RNAi strains. Metabolome analysis indicated that the adenosine triphosphate (ATP) content was higher in intact WT than cry gene mutant strains exposed to an electric field. A putative magnetoreceptor protein and UV-A/blue light photoreceptor, CRYPTOCHROME (CRY) is involved in electric field (EF) receptors in animals. The present findings constitute hitherto unknown genetic evidence of a CRY-based system that is electric field sensitive in animals.

Medical treatment using high-voltage electric potential (HELP) devices to generate an electric field (EF) is an alternative therapy commonly used in Japan. However, the mechanisms underlying potential health benefits of this therapy are still unclear. Therefore, we investigated the effect of HELP exposure (9 kV/electrode+9 kV/ electrode, 30 min) on several cytokines and hormones using enzyme-linked immunosorbent assays in plasma samples obtained from healthy human subjects before and after a single treatment session. Immunoreactive interleukin (IL)-1β and IL-6 levels were significantly downregulated following HELP exposure. Under these treatment conditions, HELP exposure did not exert on immunoreactive IL-10, IL-18, transforming growth factor-beta 1 (TGF-β1), tumor necrosis factor-alpha (TNF-α) adrenaline, serotonin, histamine, neuropeptide Y, somatostatin, insulin, or dehydroepiandrosterone sulfate (DHEAS) levels. The activation of transient receptor potential melastatin 8 (TRPM8) induces the suppression of the levels of inflammatory markers. Therefore, we further examined the in silico docking