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Huanglongbing (HLB) and citrus canker, arising from Candidatus Liberibacter asiaticus (CaLas) and Xanthomonas citri pv. Citri (Xcc), respectively, have been imposing tremendous losses to the global citrus industry. Systemic acquired resistance (SAR) has been shown to be crucial for priming defense against pathogen in citrus. Salicylic acid (SA) binding protein 2 (SABP2), which is responsible for converting methyl salicylate (MeSA) to SA, is essential for full SAR establishment. Here, we characterized the functions of four citrus SABP2 genes (CsSABP2-1, CsSABP2-1V18A, CsSABP2-2 and CsSABP2-3) against HLB and citrus canker. In vitro enzymatic assay revealed that all four proteins had MeSA esterase activities, and CsSABP2-1 and CsSABP2-1V18A has the strongest activity. Their activities were inhibited by SA except for CsSABP2-1V18A. Four genes controlled by a strong promoter 35S were induced into Wanjincheng orange (Citrus sinensis Osbeck) to generate transgenic plants overexpressing CsSABP2. Overexpressing CsSABP2 increased SA and MeSA content and CsSABP2-1V18A had the strongest action on SA. Resistance evaluation demonstrated that only CsSABP2-1V18A had significantly enhanced tolerance to HLB, although all four CsSABP2s had increased tolerance to citrus canker. The data suggested the amino acid Val-18 in the active site of CsSABP2 plays a key role in protein function. Our study emphasized that balancing the levels of SA and MeSA is crucial for regulating SAR and conferring broad-spectrum resistance to HLB and citrus canker. This finding offers valuable insights for enhancing resistance through SAR engineering. [ABSTRACT FROM AUTHOR]

AbstractIntroductionMethodsResultsDiscussionConclusionsThe toxicokinetics of methyl salicylate after unintentional or intentional ingestion of medicinal oil containing methyl salicylate has not been well studied. We aimed to characterize the trajectory of serum salicylate concentrations and to evaluate factors associated with the peak serum salicylate concentration and the time from ingestion to peak concentration.This was a retrospective cohort study of consecutive patients reported to the Hong Kong Poison Control Centre for laboratory-confirmed methyl salicylate poisoning by all local public emergency departments between 1 July 2008 and 30 June 2023. We analyzed cases with at least three serum salicylate concentrations. Multivariable generalized linear regression was used to identify factors significantly associated with the peak serum concentration and the time from ingestion to peak concentration.We included 41 patients (median age 81.0 years; 32 women and nine men). The median time from ingestion to the first peak serum salicylate concentration was 5.6 h (IQR: 3.2–10.8 h). Multiple regression showed that gastric aspiration (adjusted regression coefficient [β] − 2.50; 95% CI: −3.93 to −1.08; P = 0.001) and single-dose activated charcoal (adjusted β − 1.22; 95% CI: −2.02 to −0.42; P = 0.003) were significantly associated with a lower peak concentration, after adjusting for patient age, sex, exposure due to intentional self-harm, reported ingested dose, time from ingestion to emergency department presentation, vomiting, concurrent use of aspirin (acetylsalicylic acid) and other medications that affect gastric emptying or gastric acid secretion, blood pH, serum albumin concentration, and creatinine clearance.The serum salicylate concentration did not peak as quickly as generally believed, highlighting the importance of continued monitoring. Gastric aspiration and single-dose activated charcoal may help reduce gastrointestinal absorption, but their impact on clinical outcomes remains unclear.Given the median time of 5.6 h (IQR: 3.2–10.8 h) from ingestion to the peak salicylate concentration, gastric aspiration and single-dose activated charcoal can be considered in patients up to a few hours after medicinal oil ingestion when the airway is protected. [ABSTRACT FROM AUTHOR]

SUMMARY: Plants synthesize an array of volatile compounds, many of which serve ecological roles in attracting pollinators, deterring herbivores, and communicating with their surroundings. Methyl anthranilate (MeAA) is an anti‐herbivory defensive volatile responsible for grape aroma that is emitted by several agriculturally relevant plants, including citrus, grapes, and maize. Unlike maize, which uses a one‐step anthranilate methyltransferase (AAMT), grapes have been thought to use a two‐step pathway for MeAA biosynthesis. By mining available transcriptomics data, we identified two AAMTs in Vitis vinifera (wine grape), as well as one ortholog in "Concord" grape. Many angiosperms methylate the plant hormone salicylic acid (SA) to produce methyl salicylate, which acts as a plant‐to‐plant communication molecule. Because the Citrus sinensis (sweet orange) SA methyltransferase can methylate both anthranilate (AA) and SA, we used this enzyme to examine the molecular basis of AA activity by introducing rational mutations, which identified several active site residues that increase activity with AA. Reversing this approach, we introduced mutations that imparted activity with SA in the maize AAMT, which uncovered different active site residues from those in the citrus enzyme. Sequence and phylogenetic analysis revealed that one of the Vitis AAMTs shares an ancestor with jasmonic acid methyltransferases, similar to the AAMT from strawberry (Frageria sp.). Collectively, these data demonstrate the molecular mechanisms underpinning AA activity across methyltransferases and identify one‐step enzymes by which grapes synthesize MeAA. Significance Statement: While the two‐step pathway responsible for the biosynthesis of the grape aroma molecule, methyl anthranilate, has remained incomplete in Vitis spp., we identified two one‐step anthranilate methyltransferases in wine and one in "Concord" grapes that can methylate the tryptophan pathway intermediate anthranilate. Tracing the molecular basis of anthranilate activity in the maize and sweet orange methyltransferases uncovered distinct active site amino acids that impart substrate specificity. [ABSTRACT FROM AUTHOR]

Introduction: Oral non-steroidal anti-inflammatory drugs (NSAIDs) are effective in managing musculoskeletal pain, but often are associated with side effects. Therefore, topical anti-inflammatory drugs are considered a suitable alternative for certain patients. Objective: To determine the skin penetration capacity and anti-inflammatory efficacy of a methyl salicylate gel (MS) with turpentine essence, camphor and menthol (GelSMETAM). Methods: Human skin explants were used (three biological replicates with 3 technical replicates each were performed). To assess penetration capacity, methyl salicylate (MS) levels were quantified at different intervals using liquid chromatography with tandem mass spectrometry (LC-MS/MS). For the anti-inflammatory effect, the gene expression of major pro-inflammatory factors was quantified 30 minutes, 8 hours, and 12 hours after an impact; comparing GelSMETAM application with untreated controls. Results: The results showed that approximately 0.8% of the total applied methyl salicylate penetrated the skin with a time-dependent concentration increase. The anti-inflammatory effect demonstrated a significant reduction in the gene expression of TNFα, IL1α, IL6, IL8, and NFKβ at 30 minutes (between -39% and -53.6% compared to control), which was maintained at 8 hours except for TNFα (between -59% and -92.6%) and persisted even at 12 hours (-69.4%) for IL1α. Conclusion: The results support the hypothesis that GelSMETAM provides an effective and prolonged anti-inflammatory response, positioning it as a suitable option for the management of musculoskeletal pain. [ABSTRACT FROM AUTHOR]

The sweetpotato whitefly, Bemisia tabaci MEAM1, is one of the most devastating pests of row-crop vegetables worldwide, damaging crops directly through feeding and indirectly through the transmission of many different viruses, including the geminivirus Tomato yellow leaf curl virus (TYLCV). Y-tube olfactometer tests were conducted at different stages of TYLCV infection in tomatoes to understand how TYLCV affects B. tabaci behavior. We also recorded changes in tomato hosts' color and volatile profiles using color spectrophotometry and gas chromatography-mass spectrometry (GC-MS). We found that the infection status of B. tabaci and the infection stage of TYLCV influenced host selection, with uninfected whiteflies showing a preference for TYLCV-infected hosts, especially during the late stages of infection. Viruliferous B. tabaci attraction to visual targets significantly differed from non-viruliferous B. tabaci. Late-stage infected hosts had larger surface areas reflecting yellow-green wavelengths and higher emissions of methyl salicylate in their volatile profiles. These findings shed new light on several critical mechanisms involved in the viral manipulation of an insect vector and its economically important host. [ABSTRACT FROM AUTHOR]

An RP-HPLC method with a UV detector was developed for the simultaneous quantification of diclofenac diethylamine, methyl salicylate, and capsaicin in a pharmaceutical formulation and rabbit skin samples. The separation was achieved using a Thermo Scientific ACCLAIMTM 120 C18 column (Waltham, MA, USA, 4.6 mm × 150 mm, 5 µm). The optimized elution phase consisted of deionized water adjusted to pH = 3 using phosphoric acid mixed with acetonitrile in a 35:65% (v/v) ratio with isocratic elution. The flow rate was set at 0.7 mL/min, and the detection was performed at 205 nm and 25 °C. The method exhibits good linearity for capsaicin (0.05–70.0 µg/mL), methyl salicylate (0.05–100.0 µg/mL), and diclofenac diethylamine (0.05–100.0 µg/mL), with low LOD values (0.0249, 0.0271, and 0.0038 for capsaicin, methyl salicylate, and diclofenac diethylamine, respectively). The RSD% values were below 3.0%, indicating good precision. The overall greenness score of the method was 0.61, reflecting its environmentally friendly nature. The developed RP-HPLC method was successfully applied to analyze Omni Hot Gel® pharmaceutical formulation and rabbit skin permeation samples. [ABSTRACT FROM AUTHOR]

Non-polar clearing agents have the ability to dissolve breast adipose tissue. While fat dissolution occurs during the clearing process, careful monitoring of adipose cell membranes mis crucial due to their impact on the behavior of breast cancer cells and disease progression. Xylol is a widely employed clearing agent. However, its toxicity and prolonged clearing time necessitate the exploration of alternatives. In this context, methyl salicylate emerges as a viable substitute. It is non-polar nature facilitates efficient fat dissolution, leading to quicker tissues clarification. This study adopted a quasi-experimental method, utilizing 24 pieces of breast tissue as the sample. These tissue sample were divided into six groups according to the treatment regimen. The treatment groups involved breast tissue clearing using xylol for 2x60 minutes at room temperature, methyl salicylate for 2x30 minutes at room temperature, and methyl salicylate at 60°C for 20 minutes, 30 minutes, 2x20 minutes, and 2x30 minutes. The quality of breast tissue slides was assessed by analyzing the color contrast between the nucleus and cytoplasm using ImageJ software, along with the clarity of adipose cell cell membrane using microscopy. The results indicated that prolonged exposure at high temperature resulted in poor quality breast tissue slides. Conversely, tissue clearing with methyl salicylate for 2x30 minutes at room temperature exhibited excellent contrast between the nucleus and cytoplasm, as well as clear adipose cell membranes. Further study is warranted to explore the applicability of methyl salicylate as a clearing agent in tissues with lower fat content.

Yellow pitahaya is a tropical fruit that has gained popularity in recent years. Natural elicitors are compounds that can stimulate the resistance and quality of fruits. The objective of this study was to evaluate the effects of natural elicitors, methyl salicylate (MeSa), methyl jasmonate (JaMe), salicylic acid (SA) and oxalic acid (OA) at concentrations of 0.1 mM (MeSa and JaMe) and 5 mM (SA and OA), applied to the yellow pitahaya fruits under greenhouse conditions. After full blossom, four applications were made with a frequency of 15 days. At the time of harvest and after storage, the following variables were evaluated: firmness (whole fruit), total soluble solids (TSS), total acidity (TA), phenolics and carotenoids (in the pulp), while phenolics, carotenoids, macronutrients and micronutrients were determined in the peel. The results showed MeSa advanced the fruit maturation, according to higher TSS, lower TA and firmness than MeJa-treated fruits, for which a delayed ripening process was shown. All treatments induced a higher polyphenolic concentration during storage. Regarding the alternative use of the peel as a by-product, the application of natural elicitors significantly increased the content of polyphenols, carotenoids, macronutrients and micronutrients in the peel, especially MeSa, which can be used as a bioactive compound in the food industry. In conclusion, the results indicate that natural elicitors can be an alternative to improve the quality and shelf life of yellow pitahaya fruits. [ABSTRACT FROM AUTHOR]

Yellow pitahaya (Selenicereus megalanthus Haw.) is an exotic fruit with great potential for exportation in Ecuador. The research was carried out with the objective of evaluating the fruit growth and ripening as affected by four elicitors: salicylic acid (SA), methyl salicylate (MeSa), methyl jasmonate (MeJa), and oxalic acid (OA), all of them at 1, 5, and 10 mM concentration, compared with untreated plants (control). For each elicitor, nine plants were selected, and on each plant, three fruits were marked to follow up the growth by measuring polar and equatorial diameters. At harvest, yield (kg plant−1 and number of fruits plant−1), fruit weight, percentage of pulp and skin, total soluble solids (TSS), titratable acidity (TA), and firmness were determined. Treated plants enhanced fruit size, crop yield, and fruit weight compared with control fruits, although results depended on the elicitor tested and applied doses. The highest and lowest TSS were found in 10 mM MeSa and 5 mM MeJa-treated fruit, respectively, while the highest TA content was shown in 5 mM SA. Firmness was only enhanced in MeJa-treated fruits. Overall, results suggest that preharvest use of elicitors could modulate the pitahaya ripening and could improve quality attributes at harvest. [ABSTRACT FROM AUTHOR]

Opuntia ficus-indica has always interacted with many phytophagous insects; two of them are Dactylopius coccus and D. opuntiae. Fine cochineal (D. coccus) is produced to extract carminic acid, and D. opuntiae, or wild cochineal, is an invasive pest of O. ficus-indica in more than 20 countries around the world. Despite the economic and environmental relevance of this cactus, D. opuntiae, and D. coccus, there are few studies that have explored volatile organic compounds (VOCs) derived from the plant–insect interaction. The aim of this work was to determine the VOCs produced by D. coccus and D. opuntiae and to identify different VOCs in cladodes infested by each Dactylopius species. The VOCs (essential oils) were obtained by hydrodistillation and identified by GC-MS. A total of 66 VOCs from both Dactylopius species were identified, and 125 from the Esmeralda and Rojo Pelón cultivars infested by D. coccus and D. opuntiae, respectively, were determined. Differential VOC production due to infestation by each Dactylopius species was also found. Some changes in methyl salicylate, terpenes such as linalool, or the alcohol p-vinylguaiacol were related to Dactylopius feeding on the cladodes of their respective cultivars. Changes in these VOCs and their probable role in plant defense mechanisms should receive more attention because this knowledge could improve D. coccus rearing or its inclusion in breeding programs for D. opuntiae control in regions where it is a key pest of O. ficus-indica. [ABSTRACT FROM AUTHOR]

Simple Summary: Pest thrips species damage fruits, vegetables, field crops, and ornamental plants. They are difficult to control effectively using only insecticides. Semiochemicals produced by plants or animals are useful in integrated pest management (IPM) to reduce insecticide use in crop production. These chemicals attract arthropod pests and/or natural enemies of arthropod pests into cropping systems. Herbivore-induced plant volatiles (HIPV) or plant-produced semiochemicals such as methyl salicylate (MS) attract natural enemies and can be used as a pest management tactic. This research, conducted in Tennessee in 2018 and 2019, focused on investigating the efficacy of MS and the aggregation pheromone neryl (S)-2-methylbutanoate (NMB), an attractant of western flower thrips Frankliniella occidentalis, a severe pest of many crops, as a thrips management technique when compared with a no-lure control using sweet pepper as the crop plant. Our results show that the key species of thrips attracted to both lure treatments and the no-lure control was Frankliniellatritici, a significant pest of young cotton plants, which can also reduce the pest pressure from F. occidentalis by increasing its numbers in crop plants, banker plants, or trap crops. We discuss the possibility of using MS lure as a herbivorous thrips management practice in crop production. Herbivorous thrips that damage fruits, vegetables, field crops, and ornamentals are challenging to control using insecticides and need an integrated approach (IPM) for their management. Herbivore-induced plant volatiles (HIPVs) are semiochemical plants produced to attract natural enemies (NEs) of arthropod herbivores. Sex pheromones are animal-based semiochemicals that can attract males or females of conspecifics. The HIPV methyl salicylate (MS) is used in IPM to attract NEs. We conducted field experiments in 2018–2019 in Tennessee to study the efficacy of MS and the aggregation pheromone neryl (S)-2-methylbutanoate (NMB), which attracts Frankliniella occidentalis (FO), a dominant pest of many crops, in attracting thrips using sweet peppers. We found a significantly higher number of thrips in traps baited with MS than in the traps containing NMB when compared with a no-lure control. All treatments caught only one thrips species, Frankliniella tritici (FT), a significant pest of young cotton. It can also lower the abundance of FO in other crops. Our findings show that although FO was not found in the study location in Tennessee, traps baited with MS are suitable for managing FT and reducing FO in susceptible crops by increasing FT and attracting NEs to crop productions that use IPM-based management practices. [ABSTRACT FROM AUTHOR]

The sweetpotato whitefly, Bemisia tabaci MEAM1, is one of the most devastating pests of row-crop vegetables worldwide, damaging crops directly through feeding and indirectly through the transmission of many different viruses, including the geminivirus Tomato yellow leaf curl virus (TYLCV). Y-tube olfactometer tests were conducted at different stages of TYLCV infection in tomatoes to understand how TYLCV affects B. tabaci behavior. We also recorded changes in tomato hosts’ color and volatile profiles using color spectrophotometry and gas chromatography-mass spectrometry (GC-MS). We found that the infection status of B. tabaci and the infection stage of TYLCV influenced host selection, with uninfected whiteflies showing a preference for TYLCV-infected hosts, especially during the late stages of infection. Viruliferous B. tabaci attraction to visual targets significantly differed from non-viruliferous B. tabaci. Late-stage infected hosts had larger surface areas reflecting yellow-green wavelengths and higher emissions of methyl salicylate in their volatile profiles. These findings shed new light on several critical mechanisms involved in the viral manipulation of an insect vector and its economically important host.

The insect repellent methyl salicylate elicits excitatory responses upon interaction with CquiOR32, an odorant receptor (OR) from the southern house mosquito, Culex quinquefasciatus. By contrast, eucalyptol binds to CquiOR32 to generate electrophysiological and behavioral inhibitory responses. In an attempt to identify CquiOR32 variants displaying more robust inhibitory responses for more accurate current-voltage analysis, we sequenced 31 CquiOR32 clones. In the Xenopus oocyte recording system, CquiOR32V2/CquiOrco-expressing oocytes yielded eucalyptol-elicited outward (inhibitory) currents relatively larger than methyl salicylate-generated inward (excitatory) currents. Rescuing experiments showed that two of the amino acid substitutions in CquiOR32V2 located in a predicted transmembrane helix of the receptor are determinants of the outward/inward ratios. These findings, along with co-stimulus assays, suggest that odorant and inhibitor may bind to the same binding pocket. Current-voltage relationships obtained with standard perfusion buffer and those devoid of Na+ or Cl- indicated that both excitatory and inhibitory currents are mediated, at least in part, by cation. We then concluded that eucalyptol is an inverse agonist, which shifts the open ⇔ closed equilibrium of the receptor toward the closed conformation, thus reducing the spontaneous activity. By contrast, the binding of methyl salicylate shifts the equilibrium towards the open conformation and, consequently, leads to an increase in cation influx.

Introduction: Despite the increasing number of essential oils being reported on their potential therapeutic effects, some remain relatively unknown on their biological properties. That is the case of the essential oils obtained from copaiba (Copaifera officinalis L.), wintergreen (Gaultheria fragrantissima Wall.), everlasting (Helichrysum italicum (Roth) G.Don) and clove (Syzygium aromaticum (L.) Merr. & L.M.Perry), commonly labelled as being useful on the amelioration of conditions with an inflammatory background. Methods: To further broaden the current knowledge on the four essential oils, commercially available samples were approached on their effects upon a series of mediators that are involved on the inflammatory and oxidative response, both through in vitro cell-free and cell-based assays (5-lipoxygenase activity, lipid peroxidation, free radical and nitric oxide radical scavenging properties or tyrosinase inhibition). Results: The four oils proved to be active at some of the concentrations tested in most of the performed assays. Significant differences were found between the essential oils, S. aromaticum proving to tbe the most active, followed by G. fragrantissima against 5-lipoxygenase (5-LOX) and linoleic acid peroxidation, proving their potential use as antioxidants and anti-inflammatory agents. In fact, the IC50 value of S. aromaticum in the 5-LOX assay was 62.30 μg mL-1. Besides S. aromaticum efficiently scavenged superoxide radicals generated by xanthine/xanthine oxidase, displaying an IC50 value of 135.26 μg mL-1. The essential oil obtained from H. italicum exhibited a significant decrease in the nitric oxide levels on BV-2 cells, showing its potential as a cytoprotective agent against toxic damage. Copaiba oil ranked first as the most potent tyrosinase inhibitor, exhibiting an IC50 98.22 μg mL-1. Conclusion: More studies are needed to describe the essential oils properties, but these results confirm the potential of these essential oils as anti-inflammatory and antioxidant agents. [ABSTRACT FROM AUTHOR]

Chimaphila umbellata has been studied for almost two centuries now, with the first paper exploring the phytochemistry of the plant published in 1860. Almost all contemporary studies focus on the biotechnological advances of C. umbellata including its utilization as a natural alternative in the cosmetic, food, biofuel, and healthcare industry, with a special focus on its therapeutic uses. This literature review critically investigates the significance and applications of secondary metabolites extracted from the plant and presses on the biotechnological approaches to improve its utilization. C. umbellata is home to many industrially and medicinally important phytochemicals, the majority of which belong to phenolics, sterols, and triterpenoids. Other important compounds include 5-hydroxymethylfurfural, isohomoarbutin, and methyl salicylate (the only essential oil of the plant). Chimaphilin is the characteristic phytochemical of the plant. This review focuses on the phytochemistry of C. umbellata and digs into their chemical structures and attributes. It further discusses the challenges of working with C. umbellata including its alarming conservation status, problems with in-vitro cultivation, and research and development issues. This review concludes with recommendations based on biotechnology, bioinformatics, and their crucial interface. [ABSTRACT FROM AUTHOR]

There are many methods reported to identify and quantify volatile compounds. Many of them are based on chromatographic techniques, such as high‐performance liquid chromatography (HPLC), gas chromatography with flame ionization detector (GC‐FID), and GC coupled with mass spectrometry (GC–MS). But among all GC‐FID and GC–MS are majorly used for the detection and quantification of volatile compounds. A new gas chromatographic method has been developed for simultaneous determination of six flavouring agents [menthol, benzyl alcohol, methyl salicylate, anethole, menthol carboxamide (WS‐3) and eugenol] in herbal freshness gel toothpaste. All six flavours were separated using a DB‐WAX (Agilent) capillary GC column (30 m × 0.25 mm × 0.25 μm) and flame ionization detector (FID). This method was validated for various parameters such as precision, linearity, accuracy, stability, robustness, limit of detection, and limit of quantification as per ICH guidelines, AOAC internationals Appendix K. All the validation parameters were found to be within acceptable limits. The method was successfully applied for the quantification of all flavouring agents in toothpaste formulations. [ABSTRACT FROM AUTHOR]

Mass spectrometry experiment is an important course in instrumental chemistry. Based on our long-term practical exploration, we chose Methyl salicylate (methyl o-hydroxy Methyl benzoate) as the experimental object for mass spectrometry analysis, and compared it with its isomers 3-Hydroxybenzoic acid methyl ester, nipagin methyl ester (methyl 4-Hydroxybenzoic acid), vanillin (3-nenenebe methoxy 4-hydroxy Benzaldehyde), 2,4-dihydroxy Acetophenone and 3,4-dihydroxy Acetophenone. Through this experiment, students will master the mass spectrum pyrolysis characteristics of benzene ring, carbonyl group, Methoxy group, aldehyde group, phenols etc., and become familiar with the molecular rearrangement phenomenon of "Alpha effect of aromatic ring". This experiment is practical, interesting with moderate difficulty. It can help students to consolidate theoretical knowledge and exercise thinking, improving practical application ability. It is suitable for students major in pharmacy, chemistry, food or other related ones, and also can be used as experimental training materials for graduate students. [ABSTRACT FROM AUTHOR]

SUMMARY: Citrus production is severely threatened by the devastating Huanglongbing (HLB) disease globally. By studying and analyzing the defensive behaviors of an HLB‐tolerant citrus cultivar 'Shatangju', we discovered that citrus can sense Candidatus Liberibacter asiaticus (CLas) infection and induce immune responses against HLB, which can be further strengthened by both endogenously produced and exogenously applied methyl salicylate (MeSA). This immune circuit is turned on by an miR2977‐SAMT (encoding a citrus Salicylate [SA] O‐methyltransferase) cascade, by which CLas infection leads to more in planta MeSA production and aerial emission. We provided both transgenic and multi‐year trail evidences that MeSA is an effective community immune signal. Ambient MeSA accumulation and foliage application can effectively induce defense gene expression and significantly boost citrus performance. We also found that miRNAs are battle fields between citrus and CLas, and about 30% of the differential gene expression upon CLas infection are regulated by miRNAs. Furthermore, CLas hijacks host key processes by manipulating key citrus miRNAs, and citrus employs miRNAs that coordinately regulate defense‐related genes. Based on our results, we proposed that miRNAs and associated components are key targets for engineering or breeding resistant citrus varieties. We anticipate that MeSA‐based management, either induced expression or external application, would be a promising tool for HLB control. Significance Statement: The Great Wall not only served as a defensive barrier to ancient China, but also an intelligence transmission system. Foreign invasion alert at remote borders was signaled by rising smoke on the beacon towers and was relayed by the neighboring towers along the Great Wall. Recently researchers found a 'Great Wall' signaling system in citrus. Huanglongbing (HLB) is a devastating citrus disease causing billions of dollar losses annually. Research shows that citrus can produce and emit more methyl salicylate (MeSA) after being infected by CLas (the associated agent with HLB). MeSA is a violate that can spread through citrus growing groves and 'alarm' the uninfected citrus for the emerging threatens. This research also points out that microRNA (miRNA) is a key immune regulator in citrus. Image by: Hongwei Zhao. Due to our limited understanding of CLas pathogenicity and citrus immunity, citrus HLB causes enormous global economic losses yearly. In this investigation, we demonstrated that citrus can mount effective immune responses upon CLas infection, which is characterized by emitted community immune signal (MeSA) and citrus miRNA‐coordinated massive gene reprogramming. These two discoveries are of particular importance in the citrus industry. We could develop MeSA‐based HLB management strategies that would prime uninfected trees in the grove. In the future, we could also breed or engineer tolerant or even resistant citrus varieties and rootstocks relying on modifying critical citrus miRNAs. Our findings revealed not only citrus defensive tool kits against the HLB but also the potential of utilizing these weapons practically. [ABSTRACT FROM AUTHOR]

Avian species were historically thought to have a very limited sense of smell. However, recent studies suggest that odor plays an important role in many areas of avian life, including foraging and navigation. It has been shown that insectivorous birds can interact in a tritrophic fashion with plants and their herbivores by cuing in on herbivore-induced plant volatiles (HIPVs), important signaling compounds released by plants in response to herbivorous damage. This phenomenon has been observed in a number of plant species, including complex HIPV blends in com (Zea mays). However, in other species, simpler single-compound HIPVs can act as phytohormones, causing the release of additional HIPVs, and have also been shown to be attractive to birds. Here we test if 2 single-compound general HIPVs, methyl jasmonate and methyl salicylate, attract avian predators to insect prey on com plants using clay caterpillar models. We observed appreciable levels of bird attacks on our models across 10 experimental sites. However, we did not observe any difference between HIPV and control treatments. Interestingly, we did find that several weather variables, including wind speed and temperature, were significant or near significant predictors of avian predation activity, respectively. Together with previous studies, our results add to a nuanced picture of how birds may be using HIPVs to locate insect prey. Additionally, the potential for birds to act as a natural form of biocontrol in agricultural settings may be affected by physical location and local climate. Key words: agriculture, biocontrol, herbivory, HIPV, temperature, wind speed. On pensait historiquement que les especes aviaires avaient une olfaction tres limitee. Cependant, des etudes recentes suggerent que l'odeur joue un role important dans de nombreux domaines de la vie aviaire, y compris la recherche de nourriture et la navigation. Les oiseaux insectivores peuvent interagir a travers trois niveaux trophiques avec les plantes et leurs herbivores par les signaux des substances volatiles vegetales induites par les herbivores (HIPV), les composes de signalisation importants liberes par les plantes en reponse aux dommages causes par les herbivores. Ce phenomene a etc observe chez plusieurs especes vegetales, comme le mais (Zea mays) et ses melanges complexes d'HIPV. Chez d'autres especes, les HIPV plus simples et mono-composes peuvent agir comme des phytohormones, provoquant l'emission d'autres HIPV, qui se sont egalement averes attrayants pour les oiseaux. Ici, nous testons si deux HIPV a un seul compose, le jasmonate de methyle et le salicylate de methyle, attirent les predateurs aviaires vers les insectes proies sur les plants de mais en utilisant des modeles de chenilles en argile. Nous observames des niveaux appreciates d'attaques d'oiseaux sur nos modeles dans dix sites experimentaux. Cependant, nous ne trouvames aucune difference entre les traitements d'HIPV et les temoins. Il est interessant de noter que nous constatames que plusieurs variables meteorologiques, notamment la vitesse du vent et la temperature, etaient des predicteurs significatifs ou presque significatifs de l'activite de predation aviaire, respectivement. Nos resultats, combines a des etudes precedentes, decrivent comment les oiseaux peuvent utiliser les HIPV pour localiser les proies d'insectes. De plus, le potentiel des oiseaux a agir comme une forme naturelle de lutte biologique dans les milieux agricoles peut etre affecte par l'environnement physique et le climat local. Mots cles: agriculture, herbivorie, HIPV, lutte biologique, temperature, vitesse du vent., The role of olfaction in avian ecology has historically been ignored (Caro and Balthazart 2010, Caro et al. 2015). Only recently have odor cues been shown to play an important [...]

Stressed plants emit a variety of chemicals into the environment, leading to increased pest resistance in neighbouring plants but the genetic and molecular mechanisms of the emissions remain obscure. Recently, Gong et al. identified novel methyl salicylate (MeSA)-mediated airborne defence that confers resistance to neighbouring plants against aphids and viruses. [ABSTRACT FROM AUTHOR]

An RP-HPLC method with a UV detector was developed for the simultaneous quantification of diclofenac diethylamine, methyl salicylate, and capsaicin in a pharmaceutical formulation and rabbit skin samples. The separation was achieved using a Thermo Scientific ACCLAIMTM 120 C18 column (Waltham, MA, USA, 4.6 mm × 150 mm, 5 µm). The optimized elution phase consisted of deionized water adjusted to pH = 3 using phosphoric acid mixed with acetonitrile in a 35:65% (v/v) ratio with isocratic elution. The flow rate was set at 0.7 mL/min, and the detection was performed at 205 nm and 25 °C. The method exhibits good linearity for capsaicin (0.05–70.0 µg/mL), methyl salicylate (0.05–100.0 µg/mL), and diclofenac diethylamine (0.05–100.0 µg/mL), with low LOD values (0.0249, 0.0271, and 0.0038 for capsaicin, methyl salicylate, and diclofenac diethylamine, respectively). The RSD% values were below 3.0%, indicating good precision. The overall greenness score of the method was 0.61, reflecting its environmentally friendly nature. The developed RP-HPLC method was successfully applied to analyze Omni Hot Gel® pharmaceutical formulation and rabbit skin permeation samples.

Introduction: Despite the increasing number of essential oils being reported on their potential therapeutic effects, some remain relatively unknown on their biological properties. That is the case of the essential oils obtained from copaiba (Copaifera officinalis L.), wintergreen (Gaultheria fragrantissima Wall.), everlasting (Helichrysum italicum (Roth) G.Don) and clove (Syzygium aromaticum (L.) Merr. & L.M.Perry), commonly labelled as being useful on the amelioration of conditions with an inflammatory background.Methods: To further broaden the current knowledge on the four essential oils, commercially available samples were approached on their effects upon a series of mediators that are involved on the inflammatory and oxidative response, both through in vitro cell-free and cell-based assays (5-lipoxygenase activity, lipid peroxidation, free radical and nitric oxide radical scavenging properties or tyrosinase inhibition).Results: The four oils proved to be active at some of the concentrations tested in most of the performed assays. Significant differences were found between the essential oils, S. aromaticum proving to tbe the most active, followed by G. fragrantissima against 5-lipoxygenase (5-LOX) and linoleic acid peroxidation, proving their potential use as antioxidants and anti-inflammatory agents. In fact, the IC50 value of S. aromaticum in the 5-LOX assay was 62.30 μg mL−1. Besides S. aromaticum efficiently scavenged superoxide radicals generated by xanthine/xanthine oxidase, displaying an IC50 value of 135.26 μg mL−1. The essential oil obtained from H. italicum exhibited a significant decrease in the nitric oxide levels on BV-2 cells, showing its potential as a cytoprotective agent against toxic damage. Copaiba oil ranked first as the most potent tyrosinase inhibitor, exhibiting an IC50 98.22 μg mL−1.Conclusion: More studies are needed to describe the essential oils properties, but these results confirm the potential of these essential oils as anti-inflammatory and antioxidant agents.

Olfaction plays a crucial role for arthropods in foraging, mating, and oviposition. The odorant‐binding protein (OBP) gene is considered one of the most important olfactory genes. However, little is known about its functions in predatory mites. Here, we used Neoseiulus barkeri, an important commercialized natural pest control, to explore the chemosensory characteristics of OBP. In this study, N. barkeri was attracted by methyl salicylate (MeSA) and showed higher crawling speeds under MeSA treatment. Then, we identified and cloned an OBP gene named Nbarobp2 and analyzed its expression profiles in the predatory mite. Nbarobp2 was 663 bp, was highly expressed in larval and nymphal stages, and was significantly upregulated in N. barkeri under MeSA treatment. Nbarobp2 encoded 202 amino acid residues with a molecular weight of 23 kDa (after removing the signal peptide). Sequence comparisons revealed that the OBPs in Arachnida shared 6 conserved cysteine sites, but were distinguishable from the OBPs of Insecta on the phylogenetic tree. RNA interference, Western blotting, and binding affinity assays further proved that Nbarobp2 was involved in volatile perception in predatory mites. This study shed light on the functional characteristics of OBPs in predatory mites, providing a new insight for better biological control. [ABSTRACT FROM AUTHOR]

Objective: Immunohistochemical investigation of archival histological material is a serious problem, since long-term storage of biological tissues, most often in formalin, leads to a loss of antigenic properties. However, the biological material can also be stored in the clearing agent methyl salicylate. The aim of this study was to assess the antigenicity of the human choroid plexus after extra long-term storage in methyl salicylate. Material and Method: The study was performed on samples of fixed human choroid plexus (occasionally with attached neighboring pineal gland) stored in either methyl salicylate or paraffin blocks for 25 years. Chromogenic and fluorescence immunohistochemistry of vimentin, GFAP, type IV collagen, β-catenin, α-smooth muscle actin, von Willebrand factor, CD68, mast cell tryptase, TMEM119, and synaptophysin was carried out. Results: The storage of human choroid plexus in methyl salicylate for 25 years does not impair its histomorphology and preserves the properties of all the antigens assessed, which makes their immunohistochemical visualization possible using both light and fluorescence microscopy. Additionally, we found that long-term storage of human choroid plexus in methyl salicylate does not cause an increase in autofluorescence. Conclusion: Methyl salicylate can be recommended as a medium for long-term storage of biological tissue, as it provides excellent brain tissue preservation and retains its antigenic properties for up to 25 years.

Yellow pitahaya (Selenicereus megalanthus Haw.) is an exotic fruit with great potential for exportation in Ecuador. The research was carried out with the objective of evaluating the fruit growth and ripening as affected by four elicitors: salicylic acid (SA), methyl salicylate (MeSa), methyl jasmonate (MeJa), and oxalic acid (OA), all of them at 1, 5, and 10 mM concentration, compared with untreated plants (control). For each elicitor, nine plants were selected, and on each plant, three fruits were marked to follow up the growth by measuring polar and equatorial diameters. At harvest, yield (kg plant−1 and number of fruits plant−1), fruit weight, percentage of pulp and skin, total soluble solids (TSS), titratable acidity (TA), and firmness were determined. Treated plants enhanced fruit size, crop yield, and fruit weight compared with control fruits, although results depended on the elicitor tested and applied doses. The highest and lowest TSS were found in 10 mM MeSa and 5 mM MeJa-treated fruit, respectively, while the highest TA content was shown in 5 mM SA. Firmness was only enhanced in MeJa-treated fruits. Overall, results suggest that preharvest use of elicitors could modulate the pitahaya ripening and could improve quality attributes at harvest.

Yellow pitahaya is a tropical fruit that has gained popularity in recent years. Natural elicitors are compounds that can stimulate the resistance and quality of fruits. The objective of this study was to evaluate the effects of natural elicitors, methyl salicylate (MeSa), methyl jasmonate (JaMe), salicylic acid (SA) and oxalic acid (OA) at concentrations of 0.1 mM (MeSa and JaMe) and 5 mM (SA and OA), applied to the yellow pitahaya fruits under greenhouse conditions. After full blossom, four applications were made with a frequency of 15 days. At the time of harvest and after storage, the following variables were evaluated: firmness (whole fruit), total soluble solids (TSS), total acidity (TA), phenolics and carotenoids (in the pulp), while phenolics, carotenoids, macronutrients and micronutrients were determined in the peel. The results showed MeSa advanced the fruit maturation, according to higher TSS, lower TA and firmness than MeJa-treated fruits, for which a delayed ripening process was shown. All treatments induced a higher polyphenolic concentration during storage. Regarding the alternative use of the peel as a by-product, the application of natural elicitors significantly increased the content of polyphenols, carotenoids, macronutrients and micronutrients in the peel, especially MeSa, which can be used as a bioactive compound in the food industry. In conclusion, the results indicate that natural elicitors can be an alternative to improve the quality and shelf life of yellow pitahaya fruits.

Opuntia ficus-indica has always interacted with many phytophagous insects; two of them are Dactylopius coccus and D. opuntiae. Fine cochineal (D. coccus) is produced to extract carminic acid, and D. opuntiae, or wild cochineal, is an invasive pest of O. ficus-indica in more than 20 countries around the world. Despite the economic and environmental relevance of this cactus, D. opuntiae, and D. coccus, there are few studies that have explored volatile organic compounds (VOCs) derived from the plant–insect interaction. The aim of this work was to determine the VOCs produced by D. coccus and D. opuntiae and to identify different VOCs in cladodes infested by each Dactylopius species. The VOCs (essential oils) were obtained by hydrodistillation and identified by GC-MS. A total of 66 VOCs from both Dactylopius species were identified, and 125 from the Esmeralda and Rojo Pelón cultivars infested by D. coccus and D. opuntiae, respectively, were determined. Differential VOC production due to infestation by each Dactylopius species was also found. Some changes in methyl salicylate, terpenes such as linalool, or the alcohol p-vinylguaiacol were related to Dactylopius feeding on the cladodes of their respective cultivars. Changes in these VOCs and their probable role in plant defense mechanisms should receive more attention because this knowledge could improve D. coccus rearing or its inclusion in breeding programs for D. opuntiae control in regions where it is a key pest of O. ficus-indica.

The presence of esterase enzymes in human skin and their role in drug metabolism has been reported, but their distribution in the various skin layers and the relative contributions of those layers to metabolism is poorly defined. To gain further insight into esterase distribution, we performed in vitro skin permeation of a commercial 28.3% methyl salicylate (MeSA) cream (Metsal™) in Franz diffusion cells, using a range of human skin membranes, all from the same donor. The membranes were viable epidermis separated by a dispase II enzymatic method, heat separated epidermis, dermatomed skin, and dermis separated by a dispase II enzymatic method. Methyl salicylate and its metabolite, salicylic acid (SA), were measured by high-performance liquid chromatography. Alpha naphthyl acetate and Hematoxylin and Eosin staining provided qualitative estimations of esterase distribution in these membranes. The permeation of methyl salicylate after 24 h was similar across all membranes. Salicylic acid formation and permeation were found to be similar in dermatomed skin and dermis, suggesting dermal esterase activity. These results were supported by the staining studies, which showed strong esterase activity in the dermal–epidermal junction region of the dermis. In contrast with high staining of esterase activity in the stratum corneum and viable epidermis, minimal stained and functional esterase activity was found in heat-separated and dispase II-prepared epidermal membranes. The results are consistent with dispase II digesting hemidesmosomes, penetrating the epidermis, and affecting epidermal esterases but not those in the dermis. Accordingly, whilst the resulting dispase II-generated dermal membranes may be used for in vitro permeation tests (IVPT) involving esterase-based metabolic studies, the dispase II-generated epidermal membranes are not suitable for this purpose. [ABSTRACT FROM AUTHOR]

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Objective: Immunohistochemical investigation of archival histological material is a serious problem, since long-term storage of biological tissues, most often in formalin, leads to a loss of antigenic properties. However, the biological material can also be stored in the clearing agent methyl salicylate. The aim of this study was to assess the antigenicity of the human choroid plexus after extra long-term storage in methyl salicylate. Material and Method: The study was performed on samples of fixed human choroid plexus (occasionally with attached neighboring pineal gland) stored in either methyl salicylate or paraffin blocks for 25 years. Chromogenic and fluorescence immunohistochemistry of vimentin, GFAP, type IV collagen, β-catenin, α-smooth muscle actin, von Willebrand factor, CD68, mast cell tryptase, TMEM119, and synaptophysin was carried out. Results: The storage of human choroid plexus in methyl salicylate for 25 years does not impair its histomorphology and preserves the properties of all the antigens assessed, which makes their immunohistochemical visualization possible using both light and fluorescence microscopy. Additionally, we found that long-term storage of human choroid plexus in methyl salicylate does not cause an increase in autofluorescence. Conclusion: Methyl salicylate can be recommended as a medium for long-term storage of biological tissue, as it provides excellent brain tissue preservation and retains its antigenic properties for up to 25 years. [ABSTRACT FROM AUTHOR]

Plants use a variety of secondary metabolites to defend themselves against herbivore insects. Methyl salicylate (MeSA) is a natural plant-derived compound that has been used as a plant defence elicitor and a herbivore repellent on several crop plants. The aim of this study was to investigate the effect of MeSA treatment of Brassica rapa subsp. chinensis ('Hanakan' pak choi) on its interactions with peach potato aphids, Myzus persicae, and their natural enemy, Diaeretiella rapae. For this, we selected two concentrations of MeSA (75 mg/L and 100 mg/L). Our results showed that aphid performance was significantly reduced on plants treated with MeSA (100 mg/L). In a cage bioassay, the MeSA (100 mg/L)-treated plants showed lower adult survival and larviposition. Similarly, the MeSA (100 mg/L)-treated plants had a significantly lower aphid settlement in a settlement bioassay. In contrast, the M. persicae aphids did not show any significant difference between the MeSA (75 mg/L)-treated and control plants. In a parasitoid foraging bioassay, the parasitoid D. rapae also did not show any significant difference in the time spent on MeSA-treated and control plants. A volatile analysis showed that the MeSA treatment induced a significant change in volatile emissions, as high numbers of volatile compounds were detected from the MeSA-treated plants. Our results showed that MeSA has potential to induce defence in Brassica against M. persicae and can be utilised in developing sustainable approaches for the management of peach potato aphids. [ABSTRACT FROM AUTHOR]

Spectroscopic analysis of lawang root oil identified methyl salicylate as the major component. This result contrasts with the known composition of bark-derived lawang oil, in which eugenol is the major component. The validity of the compositional analysis was confirmed by extraction of an authentic sample of plant root tissue, and comparison with the commercially available essential oil. Lawang oil is a traditional medicine that is applied topically to reduce soreness associated with swollen joints. Spectroscopic analysis of commercial root-derived lawang oil identified methyl salicylate as the major component. This contrasts with the previous studies on the composition of bark-derived lawang oil. [ABSTRACT FROM AUTHOR]

A method for the determination of methyl salicylate in 6 kinds of cosmetics bases by gas chromatography (GC) was established, and gas chromatography-mass spectrometry (GC-MS) was used to analyze for qualitative verification. The wax-based samples were extracted with ethyl acetate-absolute ethanol, and the other bases were extracted with anhydrous ethanol, and then were determined by GC equipped with hydrogen flame ionization detector (FID). GCMS adopt full SCAN and selective ion monitoring (SIM) methods to confirm the results. Linear relationship between the mass concentration of methyl salicylate and its peak area is kept in the range of 5-500 mg/L (r=0.999 9), with the detection limit of 4 µg/g, and the quantification limit of 100 µg/g. The average recovery rate of the six bases spiked at high, medium and low concentrations are 96.9%-108.2%, and the RSDs (n=6) are all less than 2.9%. The method is used to analyze different commercially available samples. The content of methyl salicylate in one batch of the lip balm is 0.083%, which exceeds the maximum concentration recommended by Scientific Committee on Consumer Safety (SCCS). The method is fast and accurate, suitable for the qualitative and quantitative analysis of methyl salicylate in different cosmetic bases, and provides technical support for the determination and screening of methyl salicylate in cosmetics. [ABSTRACT FROM AUTHOR]