Synthesis of salicylic acid from wintergreen oil by green chemistry overcomes its cytotoxicity in keratinocyte cells and teratogenicity in zebrafish embryos.

Salicylic acid topical is used to treat variety of skin conditions. However, salicylic acid in these products is generated through industrial synthesis and has been shown to negatively impact fetal development and cause congenital abnormalities. We hypothesized that teratogenic effects reported in salicylic acid can be prevented by naturally synthesizing salicylic acid from wintergreen oil using green chemistry method. For this purpose, we investigated the effects of natural salicylic acid (NSA) synthesized from wintergreen oil using green chemistry and synthetic salicylic acid (SSA) on keratinocyte cell (HaCaT) proliferation and zebrafish embryo development. NSA structures were analyzed by 1H NMR, 13C NMR, and GC/MS methods. Percentage inhibition against HaCaT cell was determined by MTS assay. xCelligence system was used for cellular activities. Zebrafish embryos were exposed to NSA and SSA for 72 h post-fertilization. Lipid peroxidation, nitric oxide, sialic acid, glutathione-S-transferase, catalase, and superoxide dismutase were evaluated using biochemical methods. Expressions of nqO1, gfap, bdnf, vtg, egr, cyp1a, and igf2 were determined by RT-PCR as developmental indicators. MTS and RT-cell analysis showed increased cell viability by NSA, whereas SSA decreased cell viability. NSA beneficially affected zebrafish embryo development while SSA exerted deleterious effects through oxidant-antioxidant status, inflammation, and development. Results of our study showed for the first time that synthesis of salicylic acid from wintergreen oil by green chemistry overcomes its cytotoxicity in keratinocyte cells and teratogenicity in zebrafish embryos. This finding is important for drug research on safe topical applications during pregnancy, when preventing exposure to drug and chemical-derived teratogens is vital.

The combined supplementation of melatonin and salicylic acid effectively detoxifies arsenic toxicity by modulating phytochelatins and nitrogen metabolism in pepper plants.

The main objective of the study was to assess if joint application of melatonin (MT, 0.1 mM) and salicylic acid (SA 0.5 mM) could improve tolerance of pepper plants to arsenic (As) as sodium hydrogen arsenate heptahydrate (0.05 mM). The imposition of arsenic stress led to accumulation of As in roots and leaves, and increased contents of leaf proline, phytochelatins, malondialdehyde (MDA) and H2O2, but it reduced plant biomass, chlorophylls (Chl), PSII maximum efficiency (Fv/Fm) and leaf water potential. Melatonin and SA applied jointly or alone enhanced nitrogen metabolism by triggering the activities of glutamate synthase, glutamine synthetase, and nitrite reductases and nitrate. In comparison with a single treatment of MT or SA, the joint treatment of MT and SA had better impact on enhancing growth and key biological events and decreasing tissue As content. This clearly shows a cooperative function of both agents in enhancing tolerance to As-toxicity in pepper plants.

Salicylic acid antagonizes selenium phytotoxicity in rice: selenium homeostasis, oxidative stress metabolism and methylglyoxal detoxification.

We investigated the mechanistic consequences of selenium (Se)-toxicity, and its possible mitigation using salicylic acid (SA) in rice. In comparison with control, sodium selenate-exposed 'Se1' (0.5 mM) and 'Se2' (1.0 mM) plants showed accumulation of Se by 190.63 and 288.00 % in roots, 2359.42 and 2054.35 % in leaf sheaths, and 7869.91 and 9063.72 % in leaves, respectively, resulting in severe toxicity symptoms, such as growth inhibition, chlorosis, burning of leaves, and oxidative stress. In contrast, SA addition to Se-stressed plants significantly alleviated the Se-toxicity symptoms, and radically improved shoot height (28.88 %), dry biomass (34.00 %), total chlorophyll (37.51 %), soluble sugar (17.31 %) and leaf water contents (22.31 %) in 'SA + Se2' plants over 'Se2' plants. Notably, SA maintained Se-homeostasis, and decreased 'Se2'-induced oxidative stress by enhancing ascorbate level (67.75 %) and the activities of antioxidant enzymes like superoxide dismutase (20.99 %), catalase (40.97 %), glutathione peroxidase (12.26 %), and glutathione reductase (32.58 %) relative to that in 'Se2' plants. Additionally, SA protected rice plants from the deleterious effects of methylglyoxal by stimulating the activities of glyoxalase enzymes. Furthermore, SA upregulated several genes associated with reactive oxygen species (e.g. OsCuZnSOD1, OsCATB, OsGPX1 and OsAPX2) and methylglyoxal (e.g. OsGLYI-1) detoxifications. These findings unravel a decisive role of SA in alleviating Se-phytotoxicity in rice.

Establishment of auditory discrimination and detection of tinnitus induced by salicylic acid and intense tone exposure in the rat.

Rats were trained in a two lever food reinforced operant procedure to discriminate a 8000 Hz pure tone stimulus from its absence. Responding on one lever was reinforced in the presence of the tone and responding on the other lever was reinforced when the tone was absent. Frequency generalization testing yielded an inverted U-shaped function, whereas sound pressure level generalization testing yielded a continuous decrease in responding on the tone associated lever with decreasing sound pressure levels. The administration of sodium salicylic acid (150-450 mg/kg) generated responding on the tone associated lever suggesting that salicylic acid induced an experience that had commonalities with the percept of the training tone stimulus. After exposure to intense sound, responding consistent with the presence of tinnitus was achieved and Lidocaine failed to reduce tinnitus behavior. The use of a two choice design helped avoid confounding factors induced by drug induced side effects. Further, since no auditory cues were employed in the test situation the model achieves resistance to potential bias due to hearing impairment and hyperacusis. We propose that this model may be useful in detecting tinnitus.