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Thousands of electronic cigarette refill fluids are commercially available. The concentrations of nicotine and the solvents, but not the flavor chemicals, are often disclosed on product labels. The purpose of this study was to identify and quantify flavor chemicals in 39 commercial refill fluids that were previously evaluated for toxicity. Twelve flavor chemicals were identified with concentrations ≥1 mg/ml: cinnamaldehyde, menthol, benzyl alcohol, vanillin, eugenol, p -anisaldehyde, ethyl cinnamate, maltol, ethyl maltol, triacetin, benzaldehyde, and menthone. Transfer of these flavor chemicals into aerosols made at 3V and 5V was efficient (mean transfer = 98%). We produced lab-made refill fluids containing authentic standards of each flavor chemical and analyzed the toxicity of their aerosols produced at 3V and 5V using a tank Box Mod device. Over 50% of the refill fluids in our sample contained high concentrations of flavor chemicals that transferred efficiently to aerosols at concentrations that produce cytotoxicity. When tested with two types of human lung cells, the aerosols made at 5V were generally more toxic than those made at 3V. These data will be valuable for consumers, physicians, public health officials, and regulatory agencies when discussing potential health concerns relating to flavor chemicals in electronic cigarette products.

Background This study examines the impact of titanium dioxide nanoparticles (TiO 2 NPs) on gene expression associated with menthol biosynthesis and selected biochemical parameters in peppermint plants ( Mentha piperita L.). Menthol, the active ingredient in peppermint, is synthesized through various pathways involving key genes like geranyl diphosphate synthase, menthone reductase, and menthofuran synthase. Seedlings were treated with different concentrations of TiO 2 NPs (50, 100, 200, and 300 ppm) via foliar spray. After three weeks of treatment, leaf samples were gathered and kept at -70 °C for analysis. Results According to our findings, there was a significant elevation ( P  ≤ 0.05) in proline content at concentrations of 200 and 300 ppm in comparison with the control. Specifically, the highest proline level was registered at 200 ppm, reaching 259.64 ± 33.33 µg/g FW. Additionally, hydrogen peroxide and malondialdehyde content exhibited a decreasing trend following nanoparticle treatments. Catalase activity was notably affected by varying TiO 2 NP concentrations, with a significant decrease observed at 200 and 300 ppm compared to the control ( P  ≤ 0.05). Conversely, at 100 ppm, catalase activity significantly increased (11.035 ± 1.12 units/mg of protein/min). Guaiacol peroxidase activity decreased across all nanoparticle concentrations. Furthermore, RT-qPCR analysis indicated increased expression of the studied genes at 300 ppm concentration. Conclusions Hence, it can be inferred that at the transcript level, this nanoparticle exhibited efficacy in influencing the biosynthetic pathway of menthol. Graphical Abstract

Two pure natural volatile compounds, carvacrol and menthone were investigated for repellent, insecticidal, ovipositional and egg hatching inhibition activities against pulse beetle, Callosobruchus chinensis (Coleoptera: Bruchidae). Carvacrol and menthone repelled bruchid adults in choice oviposition assay. Both compounds caused toxicity, reduced oviposition potential and viability of eggs significantly when fumigated. In chronic toxicity assay, both carvacrol and menthone reduced F1 progeny production and weight loss in cowpea seeds. Reduced in grain damage was probably occurred due to inhibition of oviposition and egg hatching. Acetylcholine esterase enzyme activity was inhibited in adults when fumigated with carvacrol and menthone showing neurotic mode of action. Findings of the present study suggest that application of carvacrol and menthone can be effective in the management of C. chinensis.

The chemical composition, anti-inflammatory, cytotoxic and antioxidant activities of essential oil from leaves of Mentha piperita (MEO) grown in China were investigated. Using GC-MS analysis, the chemical composition of MEO was characterized, showing that it was mainly composed of menthol, menthone and menthy acetate. MEO exhibited potent anti-inflammatory activities in a croton oil-induced mouse ear edema model. It could also effectively inhibit nitric oxide (NO) and prostaglandin E2 (PGE2) production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. The cytotoxic effect was assessed against four human cancer cells. MEO was found to be significantly active against human lung carcinoma SPC-A1, human leukemia K562 and human gastric cancer SGC-7901 cells, with an IC50 value of 10.89, 16.16 and 38.76 µg/ml, respectively. In addition, MEO had moderate antioxidant activity. The results of this study may provide an experimental basis for further systematic research, rational development and clinical utilization of peppermint resources.

Background: Extracts of several members of the monoterpene-rich Lamiaceae sub-family Nepetoideae, including those from the Salvia (sage), Melissa (Lemon balm) and Rosmarinus (rosemary) genera, evince cognitive and mood effects in humans that are potentially related to their effects on cholinergic and GABAergic neurotransmission. To date, despite promising in vitro properties, the cognitive and mood effects of the closely related Mentha spicata (spearmint) and Mentha piperita (peppermint) remain unexplored. This study therefore assessed the human cognitive/mood effects of the M. spicata/piperita essential oil with the most promising, brain-relevant in vitro properties according to pre-trial in vitro screening. Design: Organic spearmint and peppermint (Mentha spicata/piperita) essential oils were pre-screened for neurotransmitter receptor binding and acetylcholinesterase (AChE) inhibition. In a double-blind, placebo-controlled, balanced cross-over study, 24 participants (mean age 25.2 years) consumed single doses of encapsulated placebo and 50 µL and 100 µL of the most promising essential oil (peppermint with nicotinic/GABAA receptor binding and AChE inhibitory properties, that increased calcium influx in a CAD cell neuronal model). Psychological functioning was assessed with mood scales and a range of standardised, cognitively demanding tasks pre-dose and at 1, 3 and 6 h post-dose. Results: The highest (100 µL) dose of essential oil improved performance on the cognitively demanding Rapid Visual Information Processing task (RVIP) at 1 h and 3 h post-dose and both doses attenuated fatigue and improved performance of the Serial 3 s subtraction task at 3 h post-dose. Conclusion: Peppermint (Mentha piperita) essential oil with high levels of menthol/menthone and characteristic in vitro cholinergic inhibitory, calcium regulatory and GABAA/nicotinic receptor binding properties, beneficially modulated performance on demanding cognitive tasks and attenuated the increase in mental fatigue associated with extended cognitive task performance in healthy adults. Future investigations should consider investigating higher doses.

Symbiotic associations with endophytic fungi are ecologically important for medicinal and aromatic plants. Endophytic fungi highly affect the quantity and quality of herbal products. In this study, a pot experiment was carried out in the greenhouse to investigate the interactive effects of Piriformospora indica and arbuscular mycorrhizal (AMF) inoculation on the chlorophyll fluorescence, essential oil composition, and antioxidant enzymes of peppermint under saline condition. The results showed that Fo, YNPQ, YNO, and NPQ values were obviously increased under salinity conditions, while essential oil content, chlorophyll a and b, gs, Fm, Fv, ETR, ФPSII and Fv/Fm ratio decreased by increasing salinity. In addition, salt induced the excess Na + uptake, whereas the opposite trend was observed for P and K + . The synergistic association of P . indica and AMF caused a considerable increase in the antioxidant ability, essential oil content, Fv/Fm ratio, ФPSII, and amount of P and K + uptake in salt-stressed plants. The main peppermint oil constituents, menthol, menthone, and 1,8-cineole increased considerably in inoculated plants. Besides, the applied endophytic fungi positively enhanced the ability of peppermint to alleviate the negative effect of the salinity stress.

Aromatic plants had been used since ancient times for their preservative and medicinal properties, and to impart aroma and flavor to food. Also their secondary metabolites are economically important as drugs, flavor and fragrances, pharmaceuticals, agrochemicals, dye, and pigments, pesticides, cosmetics, food additives, other industrially biochemical, and also play a major role in the adaptation of plants to their environment. Indole acetıc acid-producing rhizobacteria inoculations increase in stomatal density and level of secondary metabolite and have a synergistic effect on monoterpene biosynthesis. Bacterial inoculation significantly affected and increased the chemical composition of essential oil, citronellol, and geraniol content in rose-scented geranium; essential oil composition and total phenolic content in marigold; density, number, and size of glandular trichomes in sweet wormwood and peppermint essential oil components such as geranyl acetate, limonene, and β–pinene in coriander; oil yield and content in calendula; yield of the herb in hyssop; oxygenated compounds, essential oil content and yield, anethol and changing the chemical composition in fennel; growth, number of glandular trichomes and essential oil yield, root branching and length, and total amount of essential oil, production of monoterpenes such as pulegone, menthol, menthone, menthofuran, and terpineol content, biosynthesis of secondary metabolites in peppermint; growth and essential oil yield in marjoram; glandular hair abundance, essential oil yield, and monoterpene biosynthesis in basil; phellandrene, limonene, borneol, and campor in rosemary; carvacrol, thymol, linalool, and borneol in oregano; and α-thujene, α-pinene, α-terpinene, p-simen, β–pinene, and γ-terpinene contents and essential oil yield in summer savory. Inoculation with IAA-producing bacteria medicinal roots increased the valerenic acid in valerian, essential oil and quality in vetiver, curcumin content in turmeric alkaloid and ginsenoside content in ginseng, and inulin content in Jerusalem artichoke.

Summary Schizonepeta tenuifolia Briq. (S. tenuifolia) is a traditional Chinese medicinal plant widely used to treat colds, fever and respiratory diseases. Volatile oils including the monoterpenoids limonene, pulegone and menthone are major constituents of S. tenuifolia that determine its medicinal value and consumer popularity. A better understanding of the molecular regulatory mechanisms underlying terpenoid accumulation is needed to improve the quality and market value of S. tenuifolia. Here, we demonstrate that limonene and pulegone accumulate in S. tenuifolia under high‐light conditions, whereas menthone accumulates in the dark. Transcriptome and proteome analyses revealed that light up‐regulated both the transcriptional and protein levels of limonene synthase (LS), whereas darkness elevated pulegone reductase (PR) expression and protein abundance. An N6‐methyladenosine (m6A) dot blot assay revealed that levels of m6A modification are higher in the dark than in the light. Silencing of individual m6A writer genes resulted in higher levels of monoterpenoids such as menthone, pulegone and limonene in S. tenuifolia. We further determined that m6A modulates the content of monoterpenoids in S. tenuifolia by affecting the stability of transcripts encoding PR and LS. This study reveals a vital role for post‐transcriptional regulation mediated by the m6A modification in light intensity‐dependent terpenoid biosynthesis in S. tenuifolia and provides a strategy for genetically improving the quality of this valuable plant.

Correspondence to Dr Jenny E Ozga; JennyOzga@westat.com Background Menthol tobacco products are associated with cooling sensations in the mouth or throat and the availability of fruit-flavoured or candy-flavoured nicotine products that have cooling properties (with menthone, peppermint or the synthetic coolant WS-23, eg) has grown internationally, particularly in electronic nicotine delivery systems (ENDS)1 2 and commercial oral nicotine products.3 In addition, non-menthol cigarettes with synthetic coolants4 5 and flavour capsule cigarettes6 7 are being sold in markets where flavoured cigarette sales are prohibited.8 Due to policies that include definitions of menthol that do not comprehensively capture additives that have similar properties or evoke similar sensory effects (eg, synthetic coolants and menthol analogues),9 introduction of these novel products and additives may allow tobacco companies to circumvent restrictions on menthol tobacco manufacturing and sales. [...]the group compiled existing self-report measures related to menthol, mint and cooling-flavoured nicotine and tobacco products from the PhenX Toolkit, population-level surveys, projects led by the US Tobacco Centers of Regulatory Science (TCORS) and published literature. Population-level surveys included the Population Assessment of Tobacco and Health (PATH) Study, the International Tobacco Control Survey (both adult and youth versions), the Behavioural Risk Factor Surveillance System, the National Youth Tobacco Survey, the National Health Interview Survey, the National Survey on Drug use and Health, the National Health and Nutrition Examination Survey and the Tobacco Use Supplement to the Current Population Survey. Q1: Tobacco-flavoured Menthol Mint, wintergreen, or spearmint Ice Fruit Fruit-mint or ice-fruit combinations (such as strawberry freeze or blueberry ice)† Clove or spice Chocolate An alcoholic drink (such as wine, cognac, margarita, or other cocktails) A non-alcoholic drink (such as coffee, soda, energy drinks or other beverages) Candy, desserts or other sweets Some other flavour (specify) Q2: Yes No Q1 left ‘as is’, though ‘fruit-mint or ice-fruit combinations’ is recommended as an additional response option given that it is unclear whether someone who uses fruit-cooling flavours would indicate that