A Capsicum annuum L. seed extract exerts anti-neuroexcitotoxicity in HT22 hippocampal neurons.

The hippocampal memory deficit stands out as a primary symptom in neurodegenerative diseases, including Alzheimer's disease. While numerous therapeutic candidates have been proposed, they primarily serve to delay disease progression. Given the irreversible brain atrophy or injury associated with these conditions, current research efforts are concentrated on preventive medicine strategies. Herein, we investigated whether the extracts of Capsicum annuum L. seeds (CSE) and Capsicum annuum L. pulp (CPE) have preventive properties against glutamate-induced neuroexcitotoxicity (one of the main causes of Alzheimer's disease) in HT22 hippocampal neuronal cells. Pretreatment with CSE demonstrated significant anti-neuroexcitotoxic activity, whereas CPE did not exhibit such effects. Specifically, CSE pretreatment dose-dependently inhibited the elevation of excitotoxic elements (intracellular calcium influx and reactive oxygen species; ROS) and apoptotic elements (p53 and cleaved caspase-3). In addition, the glutamate-induced alterations of neuronal activity indicators (brain-derived neurotrophic factor; BDNF and cAMP response element-binding protein phosphorylation; CREB) were significantly attenuated by CSE treatment. We also found that luteolin is the main bioactive compound corresponding to the anti-neuroexcitotoxic effects of CSE. Our results strongly suggest that Capsicum annuum L. seeds (but not its pulp) could be candidates for neuro-protective resources especially under conditions of neuroexcitotoxicity. Its underlying mechanisms may involve the amelioration of ROS-mediated cell death and BDNF-related neuronal inactivity and luteolin would be an active compound.

Impact of induced mutation-derived genetic variability, genotype and varieties for quantitative and qualitative traits in Mentha species.

PURPOSE: The genus Mentha spp. is an aromatic herb from the family 'Lamiaceae'. It is extensively predominant in temperate and sub-temperate regions of the world. The essential oil of this species is enriched with broad aroma constituents extensively utilized in food, beverages, flavor, cosmetics, perfumery, and pharmaceutical enterprises. With the global menthol market size estimated to be worth USD 765 million in 2022, India (accompanied by China and Brazil) is the world's primary manufacturer, consumer, and exporter of Mentha oil. Despite prominent global demand, the crucial bottleneck in mint cultivation is the need for more superior commercial cultivars. Predominant vegetative propagation mode with difficulties in manual emasculation, differential blooming times, sterile/sub-sterile hybrids, and low seed viability are the primary containment in creating genetic variability by classical breeding approaches. Therefore, genetic complications encountered in conventional breeding have led the breeders to apply mutation breeding as an alternative crop improvement approach in Mentha spp. These attempts at mutation breeding have produced some distinctive mutants as genetic pools for plant breeding programs, and some novel mutant mint cultivars have been made available for commercial cultivation. CONCLUSIONS: The prime strategy in mutation-based breeding has proven an adept means of encouraging the expression of recessive genes and producing new genetic variations. The present review comprises a significant contribution of mutation breeding approaches in the development of mutant mint species and its effects on physiological variation, photosynthetic pigment, essential oil content and composition, phytochemical-mediated defense response, pathogen resistivity, and differential expression of genes related to terpenoid biogenesis. Development and diversification have led to the release of varieties, namely Todd's Mitcham, Murray Mitcham, Pranjal, Tushar, and Kukrail in M. piperita L., Mukta, and Pratik in M. cardiaca Baker, Neera in M. spicata L., Kiran in M. citrata Ehrh., and Rose mint in M. arvensis L. that have revolutionized and uplifted mint cultivation leading to economic gain by the farmers and entrepreneurs.

Ten weeks of Capsicum annuum L. extract supplementation did not change adipose tissue-derived hormones, appetite, body composition, and muscle strength when combined with resistance training in healthy untrained men: A clinical trial study.

Capsiate (CAP) is a nonpungent capsaicin analog (Capsicum annuum L. extract) that has been studied as a potential antiobesity agent. However, the interaction between chronic CAP supplementation and resistance training is not clear. The purpose of this study was to examine the changes in adipose tissue-derived hormones, body composition, appetite, and muscle strength after 10 weeks of resistance training, combined with chronic CAP supplementation in healthy untrained men. We hypothesized that CAP could induce higher benefits when combined with resistance training after 10 weeks of intervention compared to resistance training alone. Twenty-four young men (age, 22.0 ± 2.9) were randomized to either capsiate supplementation (CAP = 12 mg/day) or placebo (PL), and both groups were assigned to resistance training. Body composition, leptin and adiponectin concentrations, subjective ratings of appetite, energy intake, and exercise performance were assessed at before and after 10 weeks of progressive resistance training. There was a significant increase in body mass (P < .001), fat-free mass (CAP: 58.0 ± 7.1 vs. post, 59.7 ± 7.1 kg; PL: pre, 58.4 ± 7.3 vs. post, 59.8 ± 7.1 kg; P < .001), resting metabolic rate (CAP: pre, 1782.9 ± 160.6 vs. post, 1796.3 ± 162.0 kcal; PL: pre, 1733.0 ± 148.9 vs. post, 1750.5 ± 149.8 kcal; P < .001), maximal strength at 45 leg press (P < .001) and bench press (P < .001) in both groups, but no significant (P > .05) supplementation by training period interaction nor fat mass was observed. For subjective ratings of appetite, energy intake, leptin, and adiponectin, no significant effect of supplementation by training period interaction was observed (P > .05). In conclusion, 10 weeks of resistance training increased total body weight, muscle mass, and maximum strength in healthy untrained men; however, CAP supplementation (12 mg, 7 days per week) failed to change adipose tissue-derived hormones, appetite, body composition and muscle strength in this population. Registered under Brazilian Registry of Clinical Trials (RBR-8cz9kfq).

A contrast of emerging contaminants rac- and l-menthol toxicities to Microcystis aeruginosa through biochemical, physiological, and morphological investigations.

Fragrances rac- and l-menthol extracted from peppermint are widely used and considered as emerging contaminants recently, which are persistent in the environment. Menthol has always been considered as a safe chemical for humans, but its potential adverse ecological effects on aquatic organisms and the toxic mechanisms have not yet been fully understood. The present study aims to investigate the physiological response of Microcystis aeruginosa after exposure to the two menthol isomers, and to explore the toxic mechanisms and ecological risks of these two chemicals. Results showed that rac-menthol exhibited a hormesis effect on the cell growth, chlorophyll a and protein contents; while l-menthol showed an inhibition effect. Adenosine triphosphate (ATP) content increased significantly at day 3 and then decreased markedly at day 6 after exposure to the two chemicals. Compared with rac-menthol, l-menthol can cause damage to the antioxidant system and plasmalemma more severely, promote the production and release of microcystins-LR (MC-LR) more dramatically, upregulate the expression of MC-transportation-related gene mcyH, and induce higher apoptosis rates. Overall results revealed that the toxic effects of l-menthol on cyanobacteria were significantly greater than those of rac-menthol. The significant increase in the malondialdehyde (MDA) content and the ultrastructural characteristics of the cells indicated that the plasma membranes were damaged. Thus, further attention should be paid to the scientific use, ecological and environmental risk assessment of chiral menthol. This study will also provide a scientific basis for future water quality criteria establishment on emerging contaminants such as fragrances.

Menthol causes mitochondrial Ca2+-influx, affects structure-function relationship and cools mitochondria.

Menthol is a small bioactive compound able to cause several physiological changes and has multiple molecular targets. Therefore, cellular response against menthol is complex, and still poorly understood. In this work, we used a human osteosarcoma cell line (Saos-2) and analysed the effect of menthol, especially in terms of cellular, subcellular and molecular aspects. We demonstrate that menthol causes increased mitochondrial Ca2+ in a complex manner, which is mainly contributed by intracellular sources, including ER. Menthol also changes the Ca2+-load of individual mitochondrial particles in different conditions. Menthol increases ER-mito contact points, causes mitochondrial morphological changes, and increases mitochondrial ATP, cardiolipin, mitochondrial ROS and reduces mitochondrial membrane potential (ΔΨm). Menthol also prevents the mitochondrial quality damaged by sub-lethal and lethal doses of CCCP. In addition, menthol lowers the mitochondrial temperature within cell and also serves as a cooling agent for the isolated mitochondria in a cell free system too. Notably, menthol-induced reduction of mitochondrial temperature is observed in diverse types of cells, including neuronal, immune and cancer cells. As the higher mitochondrial temperature is a hallmark of several inflammatory, metabolic, disease and age-related disorders, we propose that menthol can serve as an active anti-aging compound against all these disorders. These findings may have relevance in case of several pharmacological and clinical applications of menthol. SIGNIFICANCE STATEMENT: Menthol is a plant-derived bioactive compound that is widely used for several physiological, behavioural, addictive, and medicinal purposes. It is a well-established "cooling and analgesic agent". However, the exact cellular and sub-cellular responses of menthol is poorly understood. In this work, we have characterized the effects of menthol on mitochondrial metabolism. Menthol regulates mitochondrial Ca2+, ATP, superoxides, cardiolipin, membrane-potential, and ER-mito contact sites. Moreover, the cooling agent menthol also cools down mitochondria and protects mitochondrial damage by certain toxins. These findings may promote use of menthol as a useful supplementary agent for anti-aging, anti-cancer, anti-inflammatory purposes where higher mitochondrial temperature is prevalent.

Beneficial Effects of Fermentation of Red Chili Pepper Using Lactococcus lactis subs. Cremoris RPG-HL-0136 in High-Fat Diet-Induced Obese Mice.

Red chili pepper is a beneficial natural spicy food that has antiobesity and antitype II diabetes effects, but it is not conducive to in-depth research as a dietary strategy to treat obesity. This study aims to investigate the beneficial effects of red chili pepper, fermented with a novel Lactococcus lactis subs. cremoris RPG-HL-0136. LC-MS/MS analysis is conducted to detect the content of capsaicin and dihydrocapsaicin, and no significant difference is observed between the nonfermented red chili pepper (NFP) (W/W) and the prepared L. lactis subs. cremoris RPG-HL-0136-fermented chili mixture (LFP). After establishing a high-fat diet-induced obese type II diabetic mouse model, the effects on weight gain, weight loss of liver and testicular fat, total cholesterol, triglyceride, fasting glucose, insulin, and homeostatic model assessment for insulin resistance in LFP were evaluated to be better than those in NFP following 10 weeks of interventions. All animal experiments were approved by the Institutional Animal Care and Use Committee of Xinxiang medical university. NFP and LFP could increase the expression of transient receptor potential vanilloid subfamily 1, peroxisome proliferator-activated receptor-alpha and caspase-2 in the high-fat mice. Compared with unfermented red chili pepper, the fermented red chili pepper complex significantly reduced LPS, tumor necrosis factor-alpha, and interleukin-6 in serum (P < .05). Intake of LFP significantly increased the expression of claudin-1 and occludin in the colon of the high-fat mice (P < .05), and there was no damage to the stomach and colon. This study provides scientific evidence that red chili pepper, fermented with L. lactis subs. cremoris RPG-HL-0136, may be beneficial for future treatment of obesity and accompanying diabetes. (IACUC.No.XYLL-20200019).

Phospholipid and menthol based nanovesicle impregnated transdermal patch for nutraceutical delivery to diminish folate and iron deficiency.

Adequate micronutrient availability is particularly important in women, children and infants. Micronutrient deficiencies are the major cause of maternal and neonatal morbidity. To overcome this, WHO recommends the use of folic acid and iron supplements for reducing anaemia and improving the health of the mother and infants. Oral intake of supplements for nutritional deficiencies are associated with gastric irritation, nausea, constipation and non-patient compliance due to associated taste. In case of absorption deficiency nutrients administered orally pass-through digestive tract unabsorbed. In the present study, we propose transdermal delivery of nutraceuticals to avoid the limitations associated with oral intake. Transdermal delivery has limited use because of the closely packed barrier of the stratum corneum that limits the permeability of molecules across skin. Here, we have used biomimetic nanovesicles impregnated in transdermal patches for delivery of folic acid and iron. Nanovesicles are prepared using an abundant component of cell membrane, phosphatidyl choline and a permeation enhancer. Further these nanovesicles are impregnated onto polyacrylate based transdermal patch.In vitrostudies have shown the ability of nanovesicles to fluidise skin lipids and penetrate into deeper skin.In vivoapplication of transdermal patches gradually increased the systemic concentration of nutraceuticals. Post application of the patch, five-fold increase in plasma folic acid concentration and 1.5-fold increase in plasma iron concertation was achieved in 6 h. Developed nanovesicles were compatible with keratinocytes and fibroblasts as testedin vitroand have the potential to enhance the cellular uptake of molecules. Skin irritation studies on human volunteers have confirmed the safety of nutraceutical loaded nanovesicles. Thus, the developed nutraceutical loaded transdermal patches provide a potential, easy to use platform for micronutrient delivery in infants and mothers.

The impact of menthol essential oil against inflammation, immunosuppression, and histopathological alterations induced by chlorpyrifos in Nile tilapia.

Chlorpyrifos (CPF) is one of the predominant water pollutants associated with inflammation and immunodepression in aquatic animals. In this study, menthol oil (MNT) impacted the immunity, antioxidative, and anti-inflammatory responses against CPF toxicity in Nile tilapia. Fish fed two diets with or without MNT and placed in four groups (control, CPF, MNT, and CPF/MNT). After 30 days, fish fed MNT displayed higher growth performance and lower FCR than CPF-intoxicated fish without feeding MNT (P < 0.05). The survival rate of fish was reduced in the CPF group without MNT feeding (P < 0.05). Blood Hb, PCV, RBCs, and WBCs were decreased in fish by CPF toxicity, while the highest Hb, PCV, RBCs, and WBCs were observed in fish fed MNT followed by those fed the control without CPF toxicity (P < 0.05). Fish fed MNT had the highest total protein, albumin, and globulin, as well as the lowest urea, bilirubin, and creatinine after 15 and 30 days. However, fish under CPF toxicity had the most inferior total protein, albumin, and globulin, as well as the highest urea, bilirubin, and creatinine among the groups (P < 0.05). The enzyme activities of ALP and ALT displayed low levels by MNT with or without CPF exposure than fish fed without MNT with or without CPF exposure after 15 and 30 days (P < 0.05). The lysozyme and phagocytic activities displayed reduced levels by CPF without MNT feeding after 15 and 30 days, while increased activities were noticed by MNT feeding without CPF toxicity followed by fish fed MNT with CPF toxicity (P < 0.05). The transcription of CAT and GPX genes displayed upregulated levels in tilapia fed MNT and exposed to CPF (P < 0.05). Also, CPF toxicity increased the transcription of the IFN-γ gene but decreased the IL-8 and IL-1ß genes. The transcription of HSP70 displayed lower levels (P < 0.05) by CPF without supplementing MNT than fish fed MNT and exposed to CPF. Histopathological analysis revealed that inflammation existed in the liver, gills, and intestine of tilapia due to CPF toxicity while MNT protected tissues from inflammation. To conclude, MNT activated the immunity, antioxidative, and anti-inflammatory responses of Nile tilapia under CPF toxicity.

Peppermint and its respective active component in diets of broiler chickens: growth performance, viability, economics, meat physicochemical properties, and carcass characteristics.

This study was conducted to investigate effects of dietary peppermint leaves and menthol supplementations on performance, survivability rate, cost economics, meat physicochemical properties, and carcass characteristics of broiler chicks. A total of 384 1-day-old, unsexed Ross 308 broiler chicks, were divided into 2 experiments. Each experiment included 192 birds that were assigned to 4 dietary treatments that included peppermint leaves at concentrations of 0, 5, 10, or 15 g/kg in the first experiment or menthol at concentrations of 0, 26, 52, or 78 mg/kg in the second experiment. Each treatment had 6 replicate pens with 8 birds. The experiments lasted for 35 D. The peppermint leaves contained 1.48% essential oil that contained 35.1% menthol, and the levels of menthol were selected based on the concentrations in peppermint leaf levels. Body weight and body-weight gain increased with the increase in dietary peppermint leaves (linear, P < 0.01) and menthol concentrations (linear, quadratic, P < 0.01) during the trial periods. In addition, the feed intake linearly increased (P < 0.01) with increasing peppermint leaves or menthol levels and, in turn, caused linear improvements (P < 0.01) in feed conversion values. Interestingly, a lower mortality rate was recorded in the supplementation groups and, therefore, a higher net return was observed. However, pH values and drip loss percentage of breast and leg muscles were not affected by either dietary peppermint levels or menthol levels. Increasing peppermint or menthol levels decreased (P < 0.001) cook-loss percentage of breast and leg muscles. On the other hand, dietary supplementation of peppermint leaves or menthol had no effect (P ≥ 0.05) on the relative weights of dressing, breast, leg, liver, heart, gizzard, spleen, or pancreas. Interestingly, abdominal fat percentage was decreased by either supplemental peppermint or menthol. Hence, the present investigation demonstrates that peppermint leaves can be used as an effective novel nutritional bio-agent up to 15 g/kg to improve the performance of broiler chicks, mainly due to its active component.

Facile formulation of near-infrared light-triggered hollow mesoporous silica nanoparticles based on mitochondria targeting for on-demand chemo/photothermal/photodynamic therapy.

The incorporation of chemo/photothermal/photodynamic therapy in subcellular organelles such as mitochondria has attracted extensive attention recently. Here, we designed mitochondria-targeted hollow mesoporous silica nanoparticles (THMSNs) loaded biocompatible phase-change material L-menthol (LM) via a facile method. Meanwhile, antitumor drug doxorubicin (DOX) and near-infrared (NIR) dye indocyanine green (ICG) approved by FDA were simultaneously encapsulated into THMSNs, denoted as THMSNs@LMDI, which showed NIR radiation triggered capacity for cancer treatment. With the mitochondria-targeted ability of triphenylphosphine, the resulting THMSNs@LMDI showed evidently improved cellular internalization and specific accumulation in mitochondria. Under NIR irradiation, the versatile ICG would be bound to simultaneously produce photodynamic and photothermal therapy. Meanwhile, in view of the solid-liquid phase transition feature of gatekeeper LM, THMSNs@LMDI provided a platform for NIR-mediated temperature-responsive DOX release. As a matter of course, these smart subcellular organelle-THMSNs could serve as an effective drug delivery platform for site-specific on-demand chemo/photothermal/photodynamic therapy of cancer.

Real-time monitoring of exhaled volatiles using atmospheric pressure chemical ionization on a compact mass spectrometer.

AIM: Breath analyses have potential to detect early signs of disease onset. Ambient ionization allows direct combination of breath gases with MS for fast, on-line analysis. Portable MS systems would facilitate field/clinic-based breath analyses. Results & methodology: Volunteers ingested peppermint oil capsules and exhaled volatile compounds were monitored over 10 h using a compact mass spectrometer. A rise and fall in exhaled menthone was observed, peaking at 60-120 min. Real-time analysis showed a gradual rise in exhaled menthone postingestion. Sensitivity was comparable to established methods, with detection in the parts per trillion range. CONCLUSION: Breath volatiles were readily analyzed on a portable mass spectrometer through a simple inlet modification. Induced changes in exhaled profiles were detectable with high sensitivity and measurable in real-time.

In vitro induction of α-pinene, pulegone, menthol, menthone and limonene in cell suspension culture of pennyroyal (Mentha pulegium).

Medicinal plants are known as important sources of secondary metabolites. Because of the economic value of pennyroyal [Mentha pulegium L. (Lamiaceae)] in food industries, propagation of this valuable plant has special importance. Plant cell suspension culture can increase some produced components. The aim of this research was performing cell culture for induction of some secondary metabolites of M. pulegium and compares it with native one. The MS medium was used for suspension culture. To investigate quantitative materials, 4 levels of yeast extract elicitor (20, 40, 60 and 80 mg/L) and salicylic acid in 4 levels (2, 4, 6 and 8 mg/L) were used. Obtained extracts were analyzed by GC-MS. Statistical analysis showed that the amount of limonene, menthone, menthol and α-pinene were more than mentioned compounds in natural plant as control. The maximum amount of this metabolites were obtained as limonene (in 60 mg/l yeast extract), menthone (in 40 mg/l yeast extract and 2 mg/l salicylic acid), menthol (in 6 mg/l salicylic acid) and α-pinene (in 4 mg/l salicylic acid) in the M. pulegium cell culture. The Pulegone was fond more in natural plants than cell culture mass. The most important secondary metabolites were increased by cell culture containing of salicylic acid and yeast extract elicitors in M. pulegume.

Glandular trichomes and essential oil characteristics of in vitro propagated Micromeria pulegium (Rochel) Benth. (Lamiaceae).

MAIN CONCLUSION: In vitro conditions and benzyladenine influenced both content and composition of micropropagated Micromeria pulegium essential oils, with pulegone and menthone being the main essential oil components. The content and chemical composition of Micromeria pulegium (Rochel) Benth. essential oils were studied in native plant material at vegetative stage and in micropropagated plants, obtained from nodal segments cultured on solid MS medium supplemented with N(6)-benzyladenine (BA) or kinetin at different concentrations, alone or in combination with indole-3-acetic acid. Shoot proliferation was achieved in all treatments, but the highest biomass production was obtained after treatment with 10 µM BA. Phytochemical analysis identified up to 21 compounds in the essential oils of wild-growing and in vitro cultivated plants, both showing very high percentages of total monoterpenoids dominated by oxygenated monoterpenes of the menthane type. Pulegone and menthone were the main essential oil components detected in both wild-growing plants (60.07 and 26.85 %, respectively) and micropropagated plants grown on either plant growth regulator-free medium (44.57 and 29.14 %, respectively) or BA-supplemented medium (50.77 and 14.45 %, respectively). The percentage of total sesquiterpenoids increased in vitro, particularly owing to sesquiterpene hydrocarbons that were not found in wild-growing plants. Differences in both content and the composition of the essential oils obtained from different samples indicated that in vitro culture conditions and plant growth regulators significantly influence the essential oils properties. In addition, the morphology and structure of M. pulegium glandular trichomes in relation to the secretory process were characterized for the first time using SEM and light microscopy, and their secretion was histochemically analyzed.

An endophyte of Picrorhiza kurroa Royle ex. Benth, producing menthol, phenylethyl alcohol and 3-hydroxypropionic acid, and other volatile organic compounds.

An endophytic fungus, PR4 was found in nature associated with the rhizome of Picrorhiza kurroa, a high altitude medicinal plant of Kashmir Himalayas. The fungus was found to inhibit the growth of several phyto-pathogens by virtue of its volatile organic compounds (VOCs). Molecular phylogeny, based on its ITS1-5.8S-ITS2 ribosomal gene sequence, revealed the identity of the fungus as Phomopsis/Diaporthe sp. This endophyte was found to produce a unique array of VOCs, particularly, menthol, phenylethyl alcohol, (+)-isomenthol, ß-phellandrene, ß-bisabolene, limonene, 3-pentanone and 1-pentanol. The purification of compounds from the culture broth of PR4 led to the isolation of 3-hydroxypropionic acid (3-HPA) as a major metabolite. This is the first report of a fungal culture producing a combination of biologically and industrially important metabolites­menthol, phenylethyl alcohol, and 3-HPA. The investigation into the monoterpene biosynthetic pathway of PR4 led to the partial characterization of isopiperitenone reductase (ipr) gene, which seems to be significantly distinct from the plant homologue. The biosynthesis of plant-like-metabolites, such as menthol, is of significant academic and industrial significance. This study indicates that PR4 is a potential candidate for upscaling of menthol, phenylethyl alcohol, and 3-HPA, as well as for understanding the menthol/monoterpene biosynthetic pathway in fungi.

Effect of gibberellic acid and calliterpenone on plant growth attributes, trichomes, essential oil biosynthesis and pathway gene expression in differential manner in Mentha arvensis L.

Extensive research is going on throughout the world to find out new molecules from natural sources to be used as plant growth promoter. Mentha arvensis L. is the main source of menthol rich essential oil used commercially in various food, pharmaceutical and other preparations. Experiments were conducted on field grown plants for understanding the effect of calliterpenone (CA), a stereo-isomer of abbeokutone, in comparison to gibberellic acid (GA3) on growth attributes, trichomes, essential oil biosynthesis and expression of some oil biosynthetic pathway genes. The exogenous application of CA (1 µM, 10 µM and 100 µM) was found to be better in improving plant biomass and stolon yield, leaf area, branching and leaf stem ratio than with counterpart GA3 at the same concentrations. CA treated plants showed higher glandular trichome number, density and diameter and also correlated with enhanced oil biogenetic capacity as revealed by feeding labeled (14)C-sucrose for 72 h to excised shoots. Semi-quantitative PCR analysis of key pathway genes revealed differential up regulation under CA treatments. Transcript level of menthol dehydrogenase/menthone reductase was found highly up regulated in CA treated plants with increased content of menthone and menthol in oil. These findings demonstrate that CA positively regulated the yields by enhanced branching and higher density of trichomes resulting into higher accumulation of essential oil. The results suggest CA as a novel plant derived diterpenoid with growth promoting action and opens up new possibilities for improving the crop yields and essential oil biosynthesis in qualitative and quantitative manner.

A natural plant growth promoter calliterpenone from a plant Callicarpa macrophylla Vahl improves the plant growth promoting effects of plant growth promoting rhizobacteria (PGPRs).

Experiments were conducted to evaluate the efficacy of calliterpenone, a natural plant growth promoter from a shrub Callicarpa macrophylla Vahl., in enhancing the growth and yield promoting effects of plant growth promoting rhizobacteria (PGPRs), in menthol mint (Mentha arvensis L).This study is based on our previous results indicating the microbial growth promotion by calliterpenone and assumption that application of calliterpenone along with PGPRs will improve the population of PGPRs resulting in higher impacts on plant growth and yield. Of the 15 PGPRs (identified as potent ones in our laboratory), 25 µl of 0.01 mM calliterpenone (8.0 µg/100 ml) was found to be useful in improving the population of nine PGPRs in culture media. The five selected strains of PGPRs exhibiting synergy with calliterpenone in enhancing growth of maize compared to PGPR or calliterpenone alone were selected and tested on two cultivars (cvs. Kosi and Kushal) of M. arvensis. Of the five strains, Bacillus subtilis P-20 (16S rDNA sequence homologous to Accession No NR027552) and B. subtilis Daz-26 (16SrDNA sequence homologuos to Accession No GU998816) were found to be highly effective in improving the herb and essential oil yield in the cultivars Kushal and Kosi respectively when co-treated with calliterpenone. The results open up the possibilities of using a natural growth promoter along with PGPRs as a bio-agri input for sustainable and organic agriculture.

Menthol and geraniol biotransformation and glycosylation capacity of Levisticum officinale hairy roots.

The biotransformation capacity of Levisticum officinale W.D.J. Koch hairy root cultures was studied by evaluating the effect of the addition of 25 mg/L menthol or geraniol on morphology, growth, and volatiles production. L. officinale hairy root cultures were maintained for 7 weeks in SH medium, in darkness at 24 degrees C and 80 r.p.m., and the substrates were added 15 days after inoculation. Growth was evaluated by measuring fresh and dry weight and by using the dissimilation method. Volatiles composition was analyzed by GC and GC-MS. Hairy roots morphology and growth were not influenced by substrate addition. No new volatiles were detected after menthol addition and, as was also the case with the control cultures, volatiles of these hairy roots were dominated by (Z)-falcarinol (1-45%), N-octanal (3-8%), palmitic acid (3-10%), and (Z)-ligustilide (2-9%). The addition of geraniol induced the production of six new volatiles: nerol/citronellol/neral (traces-15%), alpha-terpineol (0.2-3%), linalool (0.1-1.2%), and geranyl acetate (traces-2%). The relative amounts of the substrates and some of their biotransformation products decreased during the course of the experiment. Following the addition of beta-glycosidase to the remaining distillation water, analysis of the extracted volatiles showed that lovage hairy roots were able to convert both substrates and their biotransformation products into glycosidic forms. GC:gas chromatography GC-MS:gas chromatography-mass spectrometry SH:Schenk and Hildebrandt (1972) culture medium.

Ex vivo skin absorption of terpenes from Vicks VapoRub ointment.

BACKGROUND: The pharmaceutical market offers a wide range of inhalant drug products applied on the skin that contain essential oils and/or their isolated compounds, i.e. terpenes. Because there are few data concerning the skin penetration of terpenes, especially from complex carriers, the goal of this study was to determine the ex vivo skin absorption kinetics of chosen terpenes, namely eucalyptol, menthol, camphor, alpha-pinene, and beta-pinene, from the product Vicks VapoRub. MATERIAL/METHODS: Human cadaver skin was placed in a flow-through diffusion chamber and the product was applied for 15, 30, and 60 min. After the application time the skin was separated into layers using a tape-stripping technique: three fractions of stratum corneum and epidermis with dermis, and terpenes amounts in the samples were determined by gas-chromatography. RESULTS: The investigated terpenes showed different absorption characteristics related to their physicochemical properties and did not permeate through the skin into the acceptor fluid. Eucalyptol had the largest total accumulation in the stratum corneum and in the epidermis with dermis, while alpha-pinene penetrated into the skin in the smallest amount. CONCLUSIONS: The short time in which saturation of the stratum corneum with the terpenes occurred and the high accumulation of most of the investigated terpenes in the skin layers proved that these compounds easily penetrate and permeate the stratum corneum and that in vivo they may easily penetrate into the blood circulation.

The inhibition of bone resorption in rats treated with (-)-menthol is due to its metabolites.

(-)-Menthol, a monoterpene from Mentha species (Lamiaceae), has been shown to inhibit bone resorption in vivo by an unknown mechanism. In the present study, plasma and urine profiling in rats determined by GC/MS demonstrate that (-)-menthol is extensively metabolized, mainly by hydroxylation and carboxylation, and excreted in the urine, in part as glucuronides. In plasma, very low concentrations of (-)-menthol metabolites were detected after a single dose of (-)-menthol, whereas after repeated treatment, several times higher concentrations and long residence times were measured. In contrast, the elimination of unchanged (-)-menthol was increased by repeated treatment. (-)-Menthol, at concentrations found in plasma, did not inhibit bone resorption in cultured mouse calvaria (skull). However, the neutral metabolites of (-)-menthol, extracted from urine of rats fed with (-)-menthol, inhibited bone resorption in vitro, the concentrations being at plasma level or higher. These results suggest that not (-)-menthol itself, but one or several of its neutral metabolites inhibit the bone resorbing cells in vivo.

Optimization of water and nitrogen application to menthol mint (Mentha arvensis L.) through sugarcane trash mulch in a sandy loam soil of semi-arid subtropical climate.

Studies were carried out to optimize the use of water and nutrients by the crop with three moisture regimes [0.9, 1.2 and 1.5 irrigation water:cumulative pan evaporation (IW:CPE) ratios], two variables of organic mulch (control and sugarcane trash at 7 t/ha) and three levels of nitrogen (0, 100 and 200 kg/ha). Soil moisture regimes maintained at 1.2 IW:CPE ratio significantly increased the crop growth and herb and essential oil yields as compared with that of 0.9 IW:CPE ratio. The increase in herb yield due to 1.5 and 1.2 IW:CPE ratios was recorded to be 28.5% and 19%, respectively, over the irrigation given at 0.9 IW:CPE ratio, with the corresponding increase in essential oil yield to the extent of 23.5% and 15.5%. Interaction effect of moisture regimes and nitrogen rates indicated that increasing levels of irrigation at the highest level of N (200 kg/ha) improved essential oil yield of the crop. Application of N at 200 kg/ha in the mulched plots significantly enhanced the N uptake by the crop and essential oil yield over the control and 100 kg N/ha applied in the mulched/or unmulched plots and 200 kg N/ha applied in the unmulched plots. Application of organic mulch and nitrogen at 200 kg/ha improved the water use efficiency (WUE) in menthol mint crop. Higher moisture regimes maintained up to 1.2 IW:CPE ratio increased the WUE. The quality of essential oil in terms of its major constituent, menthol, improved slightly with 1.2 IW:CPE ratio as compared to 0.9 and 1.5 IW:CPE ratios at first and second harvests of the crop. It is recommended that menthol mint crop could be grown profitably by providing 16 irrigations, that is 80 cm water (based on 1.2 IW:CPE ratio) and nitrogen at 200 kg/ha in the sugarcane trash mulched plots, which could give a highest benefit:cost ratio from menthol mint cropping.