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psbA mutation (Asn266 to Thr) in Senecio vulgaris L. confers resistance to several PS II-inhibiting herbicides.

DNA sequence analysis of the psbA gene encoding the D1 protein of photosystem II (PS II), the target site of PS II-inhibiting herbicides, identified a point mutation (Asn266 to Thr) in a bromoxynil-resistant Senecio vulgaris L. population collected from peppermint fields in Oregon. Although this mutation has been previously reported in Synechocystis, this is the first report of this particular point mutation in a higher plant exhibiting resistance to PS II-inhibiting herbicides. The resistant population displayed high-level resistance to bromoxynil and terbacil (R/S ratio 10.1 and 9.3, respectively) and low-level resistance to metribuzin and hexazinone (R/S ratio 4.2 and 2.6, respectively) when compared with the susceptible population. However, the population was not resistant to the triazine herbicides atrazine and simazine or to the urea herbicide diuron. A chlorophyll fluorescence assay confirmed the resistance levels and patterns of cross-resistance of the whole-plant studies. The resistant S. vulgaris plants produced fewer seeds. Differences in cross-resistance patterns to PS II-inhibiting herbicides and the difference in fitness cost could be exploited in a weed management program.

DNA Mutational Analysis↗

Jasmine absolute (Jasminum grandiflora L.) and its mode of action on guinea-pig ileum in vitro.

Jasmine (Jasminum grandiflorum L.) is used in aromatherapy as a holistic treatment for apathy, hysteria, uterine disorders and childbirth, muscle relaxation and coughs. Its stimulant nature, on inhalation, has been shown both in animals and man. Jasmine has a spasmolytic activity on guinea-pig ileum and rat uterus in vitro. The mechanism of action of the spasmolytic activity, studied in vitro using a guinea-pig ileum smooth muscle preparation, was postsynaptic and not atropine-like. The spasmolytic effect of jasmine absolute was most likely to be mediated through cAMP, and not through cGMP. The mode of action in vitro resembled that of geranium, lavender and peppermint oils. The contradictory effect in vitro and in vivo is probably due to the solely physiological effects of jasmine absolute in vitro (producing a relaxation) compared with that in vivo, where it has a strong psychological input, producing a stimulant effect in man and enhanced movement in animals.

Animals↗

Simultaneous determination of the inhibitory potency of herbal extracts on the activity of six major cytochrome P450 enzymes using liquid chromatography/mass spectrometry and automated online extraction.

Here we describe a liquid chromatography/mass spectrometry (LC/MS) method with automated online extraction (LC/LC/MS) to simultaneously determine the in vitro inhibitory potency of herbal extracts on six major human drug-metabolising cytochrome P450 enzymes. Substrates were incubated with a commercially available mixture of CYP1A2/2C8/2C9/2C19/2D6 and 3A4 from baculovirus-infected insect cells and the resulting metabolites were quantified with LC/LC/MS using electrospray ionisation in the selected ion monitoring mode. Consistent inhibitory activities were obtained for known inhibitors and plant extracts using the enzyme/substrate cocktail and the individual enzymes/substrates. Popular herbal remedies including devil's claw root (Harpagophytum procumbens), feverfew herb (Tanacetum parthenium), fo-ti root (Polygonum multiflorum), kava-kava root (Piper methysticum), peppermint oil (Mentha piperita), eucalyptus oil (Eucalyptus globulus), red clover blossom (Trifolium pratense) and grapefruit juice (GJ; Citrus paradisi) could be identified as inhibitors of the applied CYP enzymes with IC(50) values between 20 and 1000 microg/mL.

Chemical Fractionation↗

Procedures for the isolation and quantification of the intermediates of the mevalonic acid pathway.

Procedures for the isolation and analysis of all 11 intermediates of the mevalonic acid pathway from acetyl-CoA through geranylgeranyl pyrophosphate were developed. Both acid-labile and base-labile metabolites are simultaneously extracted with good recoveries into 7 M urea at neutral pH and low temperature to minimize hydrolytic and degradative enzyme losses, and the extract is partially purified by adsorption and desorption in high yield from an anion-exchange membrane. With the use of internal standards, the pathway intermediates are subsequently separated and quantified by reversed-phase ion-pair HPLC with on-line radiodetection. Permeable secretory cells specialized for monoterpene biosynthesis were isolated from peppermint (Mentha x piperita) leaves and employed as a model system to test the analytical protocols by examining the incorporation of [14C]pyruvate, [14C]mevalonate, and [3H]isopentenyl pyrophosphate as precursors. This simple new method should be readily adaptable to a wide range of cell and tissue types that can be administered basic metabolic precursors, and should allow measurements of both flux and steady-state levels of the intermediates of the mevalonic acid pathway.

Acetyl Coenzyme A↗

Evidence that hormesis represents an "overcompensation" response to a disruption in homeostasis.

The hypothesis that growth hormesis represents an overcompensation to a disruption in homeostasis was supported in experiments assessing the effects of the synthetic plant growth inhibitor phosfon (2,4-dichlorobenzyl tributyl phosphonium chloride) on peppermint. While dose-dependent decreases in growth occurred initially in all treatment groups substantial compensation growth subsequently occurred across treatment groups. The low-dose treatment groups more than fully compensated for the initial growth decrement (20-25%), displaying a net stimulatory response by 5 weeks, whereas the high-dose treatment groups never fully compensated for the initial growth decrement (50-75%). These findings, within the context of other similar reported observations, are believed to be widely generalizable and may have important implications for biomedical fields including the design and interpretation of the bioassay especially within the context of the hazard/risk assessment process.

Dose-Response Relationship, Drug↗

Olfactory bulb cells generated in adult male golden hamsters are specifically activated by exposure to estrous females.

Two experiments were carried out to test whether cells which are born in adulthood and migrate to the olfactory bulb of adult male golden hamsters are activated during sexual behaviors, to determine the time course over which such responsiveness appears, and to ask whether activation is specific to sexual cues. In the first experiment, adult male hamsters were injected with 5'-bromodeoxyuridine (BrdU, 50mg/kg b.w.) 3 times over the course of one week in order to mark dividing cells. Ten days, three weeks, or seven weeks after the first BrdU injection, the animals were allowed to mate with an estrous female for half an hour before being sacrificed. Confocal analysis of fluorescent immunostaining of BrdU and c-Fos first revealed dual labeled cells in the olfactory bulb 3 weeks after injection of the thymidine analog. In order to determine whether the activation of these newly generated cells is specific to sexual cues, we next compared the incidence of c-Fos expression in newborn (BrdU positive) cells among male hamsters exposed to an estrous female, an aggressive male, a cotton swab containing vaginal secretion from an estrous female hamster (FHVS), a cotton swab containing peppermint, or a cotton swab containing distilled water. In the mitral and glomerular layers of the accessory olfactory bulb, animals exposed to an estrous female had significantly more double labeled cells than did those given other treatments (p < 0.01). In the mitral layer of the main bulb, animals exposed to an estrous female had a significantly higher percentage of double labeled cells than those of other groups, except those exposed to an aggressive male (p < 0.05). No double labeled cells were seen in medial preoptic area (MPOA), medial nucleus of the amygdala (Me), the bed nucleus of the stria terminalis (BNST), or the hypothalamus. Our results indicate that cells born in adulthood are more responsive to cues arising from estrous females than other stimuli, and thus may participate in sociosexual behaviors.

Animals↗

Olfactory conditioning impairment following posttraining NMDA receptor blockade in neonatal rats.

Six-day-old Sprague-Dawley rat pups were exposed to peppermint odor paired with tactile stimulation (stroking the skin with a paint brush) for twenty 10-s conditioning trials, and their olfactory preference was tested the next day. In Experiment 1, pups that had received an injection of the noncompetitive N-methyl-D-aspartate receptor antagonist MK-801 (0.1 mg/kg, i.p.) either 30 min before or immediately after conditioning spent less time over the conditioned odor than saline-treated controls. In Experiment 2, pups received an injection of either MK-801 or saline 0, 30, or 60 min after the training period. There was a reduction in the preference for the conditioned odor in the animals receiving MK-801 immediately following training, but treatment with the drug at the other intervals did not produce a performance impairment. The impairment following immediate posttraining injection occurred with either 0.05 or 0.1, but not with 0.01 mg/kg of MK-801 (Experiment 3). Experiment 4 provided control data to confirm that pups that had experienced the procedures used in Experiments 1-3 showed greater preference for the conditioned odor than did naive pups or those receiving exposure to the odor without stroking. The data indicate that immediate posttraining activation of N-methyl-D-aspartate receptors is required for normal olfactory learning in neonatal rats.

Animals↗

Inhibition of aflatoxin formation by some spices.

The effects of black pepper, cinnamon, peppermint, cumin, ginger and clove on growth and aflatoxin formation of Aspergillus flavus were studied in rice powdercorn steep (RC) medium. The effects of the first five spices were judged to be inhibition of aflatoxin formation rather than of mycelial growth. Clove completely inhibited both mycelial growth and aflatoxin formation at a concentration above 0.1%. No aflatoxin was produced when cumin and mint levels of 5% and 10% were used. Black pepper and ginger levels of 10% decreased aflatoxin formation by 100%. Higher concentrations of cinnamon, mint, cumin and ginger stimulated mycelial growth.

Aflatoxins↗

Effects of antireflux therapies on salivary function in normal humans.

Saliva has received recent attention because of its potential role in esophageal clearance and neutralization of refluxed gastric contents. Gastric antisecretory drugs or promotility drugs used to treat reflux have not been studied for their ability to affect salivary function. We conducted a double-blind randomized study of four antisecretory drugs, two promotility drugs, and placebo (PL) on 12 healthy volunteers on seven different days. Following a 6-hr fast, accumulated saliva was expectorated at least once per minute. The initial 10-min sample was discarded and a baseline 20-min sample collected. One of the following was given per os: PL, pirenzepine (PIR), 50 mg; propantheline (PRO), 30 mg; cimetidine (CIM), 300 mg; ranitidine (RAN), 150 mg; bethanechol (BET), 25 mg; or metoclopramide (MET), 10 mg. Saliva was collected for 60 min. Saliva was then stimulated (STIM) by sucking a peppermint lozenge for 30 min. Specimens were collected under oil, kept on ice, and analyzed within 30 min. Saliva flow in milliliters per minute and capacity for acid neutralization (CAN) in microequivalents acid per milliliter saliva were analyzed on all samples. The known effect of PRO to inhibit both basal saliva flow and STIM flow and CAN was seen. In contrast, the selective antimuscarinic PIR did not significantly decrease saliva flow or CAN. CIM, RAN, and MET did not significantly effect salivary function, but both CIM and RAN showed a tendency to increase CAN. Oral BET had no detectable effect on salivary function.

Adult↗

Reversible hepatic veno-occlusive disease in an infant after consumption of pyrrolizidine-containing herbal tea.

UNLABELLED: Veno-occlusive disease was diagnosed in an 18-month-old boy who had regularly consumed a herbal tea mixture since the 3rd month of life. The boy developed portal hypertension with severe ascites. Histology of the liver showed centrilobular sinusoidal congestion with perivenular bleeding and parenchymal necrosis without cirrhosis. The tea contained peppermint and what the mother thought was coltsfoot (Tussilago farfara). The parents believed the tea aided the healthy development of their child. Pharmacological analysis of the tea compounds revealed high amounts of pyrrolizidine alkaloids. Seneciphylline and the corresponding N-oxide were identified as the major components by thin-layer chromatography, mass spectrometry and NMR spectroscopy. We calculated that the child had consumed at least 60 micrograms/kg body weight per day of the toxic pyrrolizidine alkaloid mixture over 15 months. Macroscopic and microscopic analysis of the leaf material indicated that Adenostyles alliariae (Alpendost) had been erroneously gathered by the parents in place of coltsfoot. The two plants can easily be confused especially after the flowering period. The child was given conservative treatment only and recovered completely within 2 months. CONCLUSION: In all cases of veno-occlusive disease pyrrolizidine alkaloids ingestion should be excluded. The identity of collected plant material should be verified by pharmaceutically trained experts and information of composition, dosage and mode of administration should be included in guidelines for herbal preparations.

Hepatic Veno-Occlusive Disease↗

Natural occurrence of mycotoxins in different spices in Egypt.

A total of 120 different samples belonging to 24 kinds of species collected from different places at Assiut Governorate (Egypt) were examined for the natural occurrence of mycotoxins. TLC analysis of spice extracts revealed the presence of aflatoxins (8-35 micrograms/kg) in 16 samples of anise, black pepper, caraway, black cumin, fennel, peppermint, coriander and marjoram, sterigmatocystin (10-23 micrograms/kg) in ten samples of red pepper, caraway, cumin and marjoram and citrinin (8-12 micrograms/kg) in two samples of black cumin, while ochratoxin A and zearalenone could not be detected.

Aflatoxins↗

Stimulative and sedative effects of essential oils upon inhalation in mice.

This study investigated the stimulative or sedative effects of inhaling fragrant essential oils (EOs) by using a forced swimming test (FST) with mice. This behavioral test is commonly used to measure the effects of antidepressant drugs. The inhalation by mice of EOs, such as ginger oil (p<0.05), thyme oil (p<0.05), peppermint oil (p<0.05), and cypress oil (p<0.01) resulted in 5% to 22% reduction of immobility. The same results were achieved when over-agitation was artificially induced in the mice by an intraperitoneal injection of caffeine (a psycho-stimulant). In contrast, inhalation of some EOs by the mice resulted in increased immobility. To evaluate more correctly the sedative effects of EOs, the immobility of over-agitated mice induced with caffeine was ascertained after the inhalation of various EOs. Inhalation of lavender oil (p<0.01) and hyssop oil (p<0.01) increased the immobile state in mice that were treated with caffeine. The results of this study indicate that the inhalation of essential oils may induce stimulative or sedative effects in mice.

Administration, Inhalation↗

[Plant-induced toxic and allergic dermatitis (phytodermatitis)].

Herbal products are being used increasingly for medical or cosmetic purposes. Many cosmetics contain plant extracts for fragrance. Sensitizing plants in cosmetics are tea tree oil, arnica, chamomile, yarrow, citrus extracts, common ivy, aloe, lavender, peppermint, and others. However, the sensitizing potential of these plants varies. Most of the sensitizing substances are sesquiterpene lactones or terpenes. The present paper reviews the various forms of phytodermatitis, including irritant plant dermatitis, phototoxic and photo-allergic dermatitis, allergic dermatitis, and airborne contact dermatitis.

Dermatitis, Allergic Contact↗

Speciation of Mg, Mn and Zn in extracts of medicinal plants.

Aqueous extracts of birch leaves ( folium Betulae), peppermint leaves ( folium Menthae), sage leaves ( folium Salviae), valerian roots ( radix Valerianae), and dandelion roots ( radix Taraxaci) are analysed for the three essential elements magnesium, manganese and zinc. Ultrafiltration reveals that 60-100% of these metals are present as low molecular weight species (<5000 Da). Further characterisation of the low molecular weight fraction is done by using size exclusion chromatography and different detectors, namely element specific AAS detection, diode array UV-VIS detection and electrochemical detection. The similarities and differences of the plant extracts are well reflected by the respective chromatograms, and typical plant constituents can be identified by their spectral and electrochemical properties (e.g. flavonoids in Betula). Mg and Mn species are selectively detected by AAS in closely neighbouring chromatographic regions for all five samples. However, there are significant differences between the samples investigated. In all cases a good correlation exists between detection of these metals (Mg, Mn) and pulsed amperometric detection (PAD), which is selective for carbohydrates. The respective molecular weight of carbohydrate species is in the range of approximately 300-600 Da. The distribution of zinc species (detectable only in the Betula extract) is totally different from that of Mg and Mn species. For zinc, many more species are detected, and there is no correlation to carbohydrates; instead (poly)phenols are involved in zinc complexation.

Magnesium↗

Expression of a soluble flavone synthase allows the biosynthesis of phytoestrogen derivatives in Escherichia coli.

Flavones are plant secondary metabolites with potent pharmacological properties. We report the functional expression of FSI, a flavonoid 2-oxoglutarate-dependent dioxygenase-encoding flavone synthase from parsley in Escherichia coli. This expression allows the biosynthesis of various flavones from phenylpropanoid acids in recombinant E. coli strains simultaneously expressing five plant-specific flavone biosynthetic genes. The gene ensemble consists of 4CL-2 (4-coumarate:CoA ligase) and FSI (flavone synthase I) from parsley, chsA (chalcone synthase) and chiA (chalcone isomerase) from Petunia hybrida, and OMT1A (7-O-methyltransferase) from peppermint. After a 24-h cultivation, the recombinant E. coli produces significant amounts of apigenin (415 microg/l), luteolin (10 microg/l), and genkwanin (208 microg/l). The majority of the flavone products are excreted in the culture media; however, 25% is contained within the cells. The metabolic engineering strategy presented demonstrates that plant-specific flavones are successfully produced in E. coli for the first time by incorporating a soluble flavone synthase confined only in Apiaceae.

Escherichia coli↗

A new potato virus in a new lineage of picorna-like viruses.

On constructing a cDNA library for potato, 'contaminating' sequences with a significant identity to Apple latent spherical virus (ALSV) were found. Determination of the remaining genome sequence indicated the presence of a bipartite virus with an RNA1 and 2 of 7034 and 3315 nucleotides, respectively, excluding a poly(A)tail. RNA1 encodes a single polyprotein (233 kDa) and shares highest amino acid identity with ALSV at 65%. Conserved amino acid motifs typical for helicase, protease and RNA-dependent polymerase (RdRp) functions are present. RNA2 encodes a single polyprotein (106 kDa) with amino acid identities to the flat apple isolate of Cherry rasp leaf virus (CRLV-FA) (97%) and ALSV (70%), suggesting this is a potato strain of CRLV (CRLV-pot). Phylogenetic analysis using the RdRp region shows that this virus falls within a group separate from the Comoviridae that includes members of the Sequiviridae and the taxonomically unassigned viruses ALSV, Strawberry mottle virus, Satsuma dwarf virus and Navel orange infectious mottling virus. Other regions of the genome have highest identities with both plant and animal infecting members of the picorna-like virus superfamily. The evolutionary context of CRLV-pot and related viruses is discussed. Similar viral sequences from an EST library of peppermint are also analysed.

5' Untranslated Regions↗

Screening of the antibacterial effects of a variety of essential oils on respiratory tract pathogens, using a modified dilution assay method.

The purpose of this study was to examine the antibacterial effects of a wide variety of essential oils on major respiratory tract pathogens. The antibacterial activity of 14 essential oils and their major components was evaluated by agar-plate dilution assay under sealed conditions, with agar used as a stabilizer for homogeneous dispersion. Of the selected strains of four major bacteria causing respiratory tract infection, Haemophilus influenzae was most susceptible to the essential oils, followed by Streptococcus pneumoniae and Streptococcus pyogenes. Staphylococcus aureus was less susceptible. No cross-resistance was observed between penicillin-sensitive and penicillin-resistant S. pneumoniae. Escherichia coli, used as a control bacterium, showed the lowest susceptibility. Essential oils containing aldehyde or phenol as a major component showed the highest antibacterial activity, followed by the essential oils containing terpene alcohols. Other essential oils, containing terpene ketone, or ether, had much weaker activity, and an oil containing terpene hydrocarbon was inactive. Based on these findings, thyme (wild, red, and geraniol types), cinnamon bark, lemongrass, perilla, and peppermint oils were selected for further evaluation of their effects on respiratory tract infection.

Agar↗

Demonstration that limonene is the first cyclic intermediate in the biosynthesis of oxygenated p-menthane monoterpenes in Mentha piperita and other Mentha species.

The volatile oil of mature Mentha piperita (peppermint) leaves contains as major components the oxygenated p-menthane monoterpenes l-menthol (47%) and l-menthone (24%) as well as very low levels of the monoterpene olefins limonene (1%) and terpinolene (0.1%), which are considered to be probable precursors of the oxygenated derivatives. Immature leaves, which are actively synthesizing monoterpenes, produce an oil with comparatively higher levels of limonene (approximately 3%), and isolation of the pure olefin showed this compound to consist of approximately 80% of the l-(4S)-enantiomer and approximately 20% of the d-(4R)-enantiomer. The time course of incorporation of [U-14C]sucrose into the monoterpenes of M. piperita shoot tips was consistent with the initial formation of limonene and its subsequent conversion to menthone via pulegone. d,l-[9-3H]Limonene and [9,10-3H]terpinolene were prepared and tested directly as precursors of oxygenated p-menthane monoterpenes in M. piperita shoot tips. Limonene was readily incorporated into pulegone, menthone, and other oxygenated derivatives, whereas terpinolene was not appreciably incorporated into these compounds. Similarly, d,l-[9-3H]limonene was specifically incorporated into pulegone in Mentha pulegium and into the C-2-oxygenated derivative carvone in Mentha spicata, confirming the role of this olefin as the essential precursor of oxygenated p-menthane monoterpenes. Soluble enzyme preparations from the epidermis of immature M. piperita leaves converted the acyclic terpenoid precursor [1-3H]geranyl pyrophosphate to limonene as the major cyclic product, providing a further indication that this olefin plays a central role in the formation of oxygenated monoterpenes in Mentha. No free intermediates were detected in the cyclization of geranyl pyrophosphate to limonene, suggesting that the olefin is the first cyclic intermediate to arise in the pathway, and resolution of the biosynthetic limonene, by crystallization of the derived d- and l-carvoximes, indicated an enantiomer mixture nearly identical to that isolated from the leaf oil.

Cell-Free System↗