Jun-Hyung Tak

Comparative and synergistic activity of Rosmarinus officinalis L. essential oil constituents against the larvae and an ovarian cell line of the cabbage looper, Trichoplusia ni (Lepidoptera: Noctuidae)

BACKGROUND Plant essential oils are usually complex mixtures, and many factors can affect their chemical composition. To identify relationships between the composition and bioactivity of the constituents, comparative and synergistic interactions of the major constituents of rosemary essential oil were evaluated against third-instar larvae and an ovarian cell line of the cabbage looper, Trichoplusia ni, via different methods of application. RESULTS The major constituents of the rosemary oil we used were 1,8-cineole, (±)-camphor, (+)-α-pinene and camphene. Via topical application to larvae, 1,8-cineole was identified as the major active compound, whereas via fumigation, 1,8-cineole and (±)-camphor, and in a cytotoxicity assay, (+)-α-pinene, were determined to be the major active principles. Several combinations of these constituents exhibited synergistic insecticidal activities when topically applied, particularly among combinations of three major constituents, (±)-camphor, (+)-α-pinene and camphene. A binary mixture of 1,8-cineole and (±)-camphor showed enhanced activity, with a synergy ratio of 1.72. CONCLUSION Based on our results, the insecticidal activity of rosemary oil appears to be a consequence of the synergistic interaction between 1,8-cineole and (±)-camphor, and (±)-camphor should be considered a promising synergizing agent.

Contact, fumigant, and cytotoxic activities of thyme and lemongrass essential oils against larvae and an ovarian cell line of the cabbage looper, Trichoplusia ni

The chemical composition of a plant essential oil can be affected by many environmental and biological factors. Understanding the role of individual constituents as well as their interactions to the overall insecticidal bioactivity is prerequisite to the use of essential oils as an alternative to conventional insecticides. In the present study, the chemical compositions of plant essential oils obtained from Thymus vulgaris (thyme) and Cymbopogon citratus (lemongrass) were analyzed by gas chromatography/mass spectrometry, and the insecticidal and cytotoxic activities of individual constituents were evaluated against third instar larvae and an ovarian cell line of Trichoplusia ni. Thymol was the most abundant compound in thyme oil and the primary active constituent in contact and cytotoxicity tests, whereas p-cymene was the most effective compound for fumigant toxicity. In lemongrass oil, citral was identified as the major active and most abundant constituent. A weak correlation between insecticidal activity and cytotoxicity was observed, indicating limitation of the latter as a screening tool for novel insecticides. Although the evaporation of thymol was enhanced at higher temperatures, its contribution to fumigant activity was limited.

Enhanced cuticular penetration as the mechanism for synergy of insecticidal constituents of rosemary essential oil in Trichoplusia ni

Synergistic interactions between constituents of essential oils have been reported for several areas of research. In the present study, mechanisms that could explain the synergistic action of the two major insecticidal constituents of rosemary oil, 1,8-cineole and camphor against the cabbage looper, Trichoplusia ni were investigated. 1,8-Cineole was more toxic than camphor when applied topically to larvae, and when coadministered in their ratio naturally occurring in rosemary oil, the binary mixture was synergistic. However, when injected directly into larvae, camphor was more toxic than 1,8-cineole. GC-MS analyses showed that penetration of topically-applied camphor was significantly enhanced when it was mixed with 1,8-cineole in the natural ratio. A bioassay combining injection and topical application methods confirmed the increased penetration of both compounds when mixed, showing the same bioactivity as seen for higher amounts applied individually. Lowered surface tension as well as increased solubility of camphor by 1,8-cineole, along with the interaction between 1,8-cineole and the lipid layer of the insect’s cuticle may explain the enhanced penetration of camphor. Because of the similarities in biological function of animal and microbial membranes, our finding has potential for application in other fields of study.

Metabolism of citral, the major constituent of lemongrass oil, in the cabbage looper, Trichoplusia ni, and effects of enzyme inhibitors on toxicity and metabolism

Although screening for new and reliable sources of botanical insecticides remains important, finding ways to improve the efficacy of those already in use through better understanding of their modes-of-action or metabolic pathways, or by improving formulations, deserves greater attention as the latter may present lesser regulation hurdles. Metabolic processing of citral (a combination of the stereoisomers geranial and neral), a main constituent of lemongrass (Cymbopogon citratus) essential oil has not been previously examined in insects. To address this, we investigated insecticidal activities of lemongrass oil and citral, as well as the metabolism of citral in larvae of the cabbage looper, Trichoplusia ni, in associations with well-known enzyme inhibitors. Among the inhibitors tested, piperonyl butoxide showed the highest increase in toxicity followed by triphenyl phosphate, but no synergistic interaction between the inhibitors was observed. Topical application of citral to fifth instar larvae produced mild reductions in food consumption, and frass analysis after 24h revealed geranic acid (99.7%) and neric acid (98.8%) as major metabolites of citral. Neither citral nor any other metabolites were found following in vivo analysis of larvae after 24h, and no significant effect of enzyme inhibitors was observed on diet consumption or citral metabolism.

Penetration-enhancement underlies synergy of plant essential oil terpenoids as insecticides in the cabbage looper, Trichoplusia ni

Many plant essential oils and their terpenoid constituents possess bioactivities including insecticidal activity, and they sometimes act synergistically when mixed. Although several hypotheses for this have been proposed, the underlying mechanism has not been fully elucidated thus far. In the present study, we report that in larvae of the cabbage looper, Trichoplusia ni, most synergistic or antagonistic insecticidal activities among mixtures of plant essential oil constituents are pharmacokinetic effects, owing to changes in solubility as well as spreadability on a wax layer. Among the major constituents of rosemary (Rosmarinus officinalis) oil, in vitro analysis revealed up to a 19-fold increase in penetration of camphor in a binary mixture with 1,8-cineole through the larval integument, suggesting increased penetration as the major mechanism for synergy. A total of 138 synergistic or antagonistic interactions among 39 compounds were identified in binary mixtures via topical application, and these were highly correlated to changes in surface tension as measured by contact angle of the mixtures on a beeswax layer. Among compounds tested, trans-anethole alone showed evidence of internal synergy, whereas most of remaining synergistic or antagonistic combinations among the three most active compounds were identified as penetration-related interactions, confirmed via a divided-application bioassay.

Effects of rosemary, thyme and lemongrass oils and their major constituents on detoxifying enzyme activity and insecticidal activity in Trichoplusia ni

Although there have been many reports on the synergistic interactions among the major constituents of plant essential oils regarding insecticidal activity, their underlying mechanism of synergy is poorly understood. In our previous studies, we found each of the two most abundant constituents of rosemary (Rosmarinus officinalis L.), thyme (Thymus vulgaris L.) and lemongrass (Cymbopogon citratus Stapf.) essential oils can be synergistic against the larvae of the cabbage looper, Trichoplusia ni at their natural proportion or equivalent blending ratios. In the present study, we investigated whether the enhanced toxicity between the major constituents could be the result of inhibited enzyme activity of cytochrome P450s, general esterases or glutathione S-transferases which are highly related to the development of insecticide resistance. Overall, although some combinations showed mild inhibitory activity, at least for these essential oils and their major constituents, inhibition of detoxication enzyme activity is unlikely to be a direct cause of increased toxicity in the cabbage looper. The results point to other factors, such as multiple modes-of-action or enhanced penetration through the cuticular layer, playing important roles in the elevated insecticidal activity. Moreover, application of enzyme inhibitors sometimes resulted in decreased activity when mixed with the target compounds, but these antagonistic interactions disappeared when they were applied separately, suggesting that the enzyme inhibitors can sometimes influence the penetrations of toxicants.

Identification of a Chrysanthemic Ester as an Apolipoprotein E Inducer in Astrocytes.

The apolipoprotein E (APOE) gene is the most highly associated susceptibility locus for late onset Alzheimer’s Disease (AD), and augmenting the beneficial physiological functions of apoE is a proposed therapeutic strategy. In a high throughput phenotypic screen for small molecules that enhance apoE secretion from human CCF-STTG1 astrocytoma cells, we show the chrysanthemic ester 82879 robustly increases expressed apoE up to 9.4-fold and secreted apoE up to 6-fold and is associated with increased total cholesterol in conditioned media. Compound 82879 is unique as structural analogues, including pyrethroid esters, show no effect on apoE expression or secretion. 82879 also stimulates liver x receptor (LXR) target genes including ATP binding cassette A1 (ABCA1), LXRα and inducible degrader of low density lipoprotein receptor (IDOL) at both mRNA and protein levels. In particular, the lipid transporter ABCA1 was increased by up to 10.6-fold upon 82879 treatment. The findings from CCF-STTG1 cells were confirmed in primary human astrocytes from three donors, where increased apoE and ABCA1 was observed along with elevated secretion of high-density lipoprotein (HDL)-like apoE particles. Nuclear receptor transactivation assays revealed modest direct LXR agonism by compound 82879, yet 10 μM of 82879 significantly upregulated apoE mRNA in mouse embryonic fibroblasts (MEFs) depleted of both LXRα and LXRβ, demonstrating that 82879 can also induce apoE expression independent of LXR transactivation. By contrast, deletion of LXRs in MEFs completely blocked mRNA changes in ABCA1 even at 10 μM of 82879, indicating the ability of 82879 to stimulate ABCA1 expression is entirely dependent on LXR transactivation. Taken together, compound 82879 is a novel chrysanthemic ester capable of modulating apoE secretion as well as apoE-associated lipid metabolic pathways in astrocytes, which is structurally and mechanistically distinct from known LXR agonists.

Essential Oils for Arthropod Pest Management in Agricultural Production Systems

Abstract Essential oils are volatile compounds extracted from mostly leaves and nonwoody parts of plants. Extracted essential oils possess scent-producing compounds including monoterpenes, sesquiterpenes, phenols, oxides, esters, aldehydes, and ketones. Plants use these compounds for protection from various plant-feeding arthropods (e.g., insects and mites). Modes of action of essential oils vary include neurotoxicity, regulation of insect growth, deterioration of the waxy layers of the insect cuticle, impeding digestive enzymes, and inhibition of glutathione-S-transferase. Currently, only a few essential oil-based pesticides have been commercialized for agricultural use although many essential oils have been found to be very effective as pesticides. Because essential oils are environmentally friendly they can play an important role as pesticides in integrated pest management and organic farming. In this chapter, modes of action of essential oils, essential oils for arthropod pest management, and advantages and disadvantages of essential oils as pesticides are presented and discussed.