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Bioactivity of selected plant essential oils against Listeria monocytogenes.

Ninety-three different commercial essential oils were screened for activity against 20 Listeria monocytogenes strains in vitro and the results correlated against the actual chemical composition of each oil. There was a substantial difference in the activity between different essential oils as expected, but there was also a difference in activity between different samples of the same essential oil. Strong anti-Listeria activity was often correlated with essential oils containing a high percentage of monoterpenes, eugenol, cinnamaldehyde, thymol, and sometimes with citronellol, limonene and geraniol. However, as there was often no correlation between the anti-Listeria activity and the main chemical components, it is possible that either there is a more complex relationship with the chemical composition (which includes the minor components) or that substantial adulteration had occurred in some essential oil samples.

Listeria monocytogenes↗

Combined action of S-carvone and mild heat treatment on Listeria monocytogenes Scott A.

The combined action of the plant-derived volatile, S-carvone, and mild heat treatment on the food-borne pathogen, Listeria monocytogenes, was evaluated. The viability of exponential phase cultures grown at 8 degrees C could be reduced by 1.3 log units after exposure to S-carvone (5 mmol l-1) for 30 min at 45 degrees C, while individual treatment with S-carvone or exposure to 45 degrees C for 30 min did not result in a loss in viability. Other plant-derived volatiles, namely carvacrol, cinnamaldehyde, thymol and decanal, were also found to reduce the viability of L. monocytogenes in combination with the same mild heat treatment at concentrations of 1.75 mmol l-1, 2.5 mmol l-1, 1.5 mmol l-1 and 2 mmol l-1, respectively. These findings show that essential oil compounds can play an important role in minimally processed foods, and can be used in the concept of Hurdle Technology to reduce the intensity of heat treatment or other individual hurdles.

Colony Count, Microbial↗

Antifungal activity of essential oils evaluated by two different application techniques against rye bread spoilage fungi.

AIMS: To study how antifungal activity of natural essential oils depends on the assay method used. METHODS AND RESULTS: Oils of bay, cinnamon leaf, clove, lemongrass, mustard, orange, sage, thyme and two rosemary oils were tested by two methods: (1) a rye bread-based agar medium was supplemented with 100 and 250 microl l-1 essential oil and (2) real rye bread was exposed to 136 and 272 microl l-1 volatile oil in air. Rye bread spoilage fungi were used for testing. Method 1 proved thyme oil to be the overall best growth inhibitor, followed by clove and cinnamon. On the contrary, orange, sage and rosemary oils had very limited effects. Mustard and lemongrass were the most effective oils by the volatile method, and orange, sage and one rosemary showed some effects. Oil compositions were analysed by gas chromatography-mass spectrography. CONCLUSIONS: Antifungal effects of the essential oils depended on the application method. Larger phenolic compounds such as thymol and eugenol (thyme, cinnamon and clove) had best effect applied directly to medium, whereas smaller compounds such as allyl isothiocyanate and citral (mustard and lemongrass) were most efficient when added as volatiles. SIGNIFICANCE AND IMPACT OF THE STUDY: This study proves that the method used for screening essential oils as potential antimicrobials should correspond with the application sought.

Antifungal Agents↗

Chemical analysis and antifungal activity of the essential oil of Calea clematidea.

The chemical composition of the essential oils of Calea clematidea Baker obtained by hydrodistillation of the leaves and flowers was analysed by GC and GC/MS and the oils were assayed for their antifungal activities. The essential oil of the leaves showed a high content of a new natural epoxy terpenoid, named clemateol (ca. 70 %), with minor amounts of o-vanillin (6.5 %), spathulenol (4.2 %), alpha-terpinene (4.0 %), germacrene B (2.9 %), yomogi alcohol (1.8 %), ( E)-caryophylene (1.7 %), m-cymenene (1.6 %), and alpha-gurjunene (1.5 %), while the essential oil of the flowers was characterized by a higher content of thymol methyl ether (ca. 80 %), with minor amounts of clemateol (4.8 %) and o-cymene (4.7 %). The antimicrobial activity of the oils was also evaluated against dermatophytes for their possible use in pharmaceutical preparations for topical applications. The oil of the leaves (MIC > 3.57 mg/ml), clemateol (MIC > 1.52 mg/ml), and the alcohol 2 (MIC > 2.82 mg/ml) showed a moderate antifungal activity against Trichophyton tonsurans, Trichophyton rubrum, Trichophyton menthagrophytes var. i nterdigitale, Epidermophyton floccosum, Microsporum gypseum, Microsporum canis and Microsporum nanum.

Antifungal Agents↗

[Micro-computed tomography of the vasculature in parenchymal organs and lung alveoli].

Micro-CT has become a powerful technique in non-destructive 3D imaging and morphometric analysis. First results were limited to the investigation of osteoporosis in cancellous bone. But the availability of systems with almost microscopic resolution and sufficient soft tissue contrast has opened up entirely new applications for laboratory investigation of blood vessels and soft tissues. This article gives an overview of micro-CT technology and the potential of three-dimensional imaging of the vessel wall and soft-tissue architecture imaging in different organs using different contrast perfusion and staining techniques. Micro-CT provides quantitative information on human plaque morphology equivalent to histomorphometric analysis. Based on differences in grey-scale attenuations, micro-CT also correctly identifies atherosclerotic lesions that are histologically classified as fibrous plaques, calcified lesions, fibroatheroma, and lipid rich lesions. Micro-CT is a promising method to visualize the architecture of the renal vasculature and, importantly, to separate cortex and medulla for the visualization of glomeruli and their afferent and efferent arterioles. Micro-CT can determine the vascular surface in a defined placental volume. Combining of micro-CT data and total placental volume enables an estimation of the approximate surface of the placental vasculature. The diameter of opacified vessels in the investigated samples ranged from 2 mm (chorion plate artery) to 14 micro m (smallest vessel diameter, terminal loop). Recognizing that lung parenchyma can only be visualized if the alveoli are completely expanded and the contrast of the thin alveolar walls is enhanced, we tested two preparation methods: (1) fixation of lung tissue with formalin vapour and staining with silver nitrate, and (2) intravenous injection of a barium sulfate-gelatine-thymol mixture in vivo in the anesthetized animal. We evaluated the ability of this mixture to enter the pulmonary microcirculation and the technical feasibility of micro-CT to assess lung micro-architecture.

Animals↗

Antifungal activities of essential oils from Thymus quinquecostatus and T. magnus.

The antifungal activities of essential oils from Thymus quinquecostatus and T. magnus, which are species native to Korea, were evaluated against seven common pathogenic fungi. Additionally, the effects of the oils together with ketoconazole were tested by the checkerboard titer test. Both of the two Thymus oils showed significant inhibition of the tested fungi, with minimal inhibitor concentrations (MICs) and minimal fungicidal concentrations (MFCs) in the range of 0.04-0.39 mg/mL and 0.19-0.78 mg/mL, respectively. The two Thymus oils, and thymol as well, exhibited synergism with ketoconazole against Trichophyton rubrum, which showed the highest susceptibility to these oils.

Antifungal Agents↗

In vivo antimalarial activity of essential oils from Cymbopogon citratus and Ocimum gratissimum on mice infected with Plasmodium berghei.

The essential oils obtained by hydrodistillation from fresh leaves of Cymbopogon citratus and Ocimum gratissimum growing in Cameroon were analyzed by GC and GC/MS. The main constituents of the oil of Ocimum gratissimum were gamma-terpinene (21.9 %), beta-phellandrene (21.1 %), limonene (11.4 %) and thymol (11.2 %), while the oil of Cymbopogon citratus contained geranial (32.8 %), neral (29.0 %), myrcene (16.2 %) and beta-pinene (10.5 %). The effects of these oils on the growth of Plasmodium berghei were investigated. Both oils showed significant antimalarial activities in the four-day suppressive in vivo test in mice. At concentrations of 200, 300 and 500 mg/kg of mouse per day, the essential oil of C. citratus produced the highest activity with the respective percentages of suppression of parasitaemia: 62.1 %, 81.7 % and 86.6 %. The corresponding values for the oil of O. gratissimum at the same concentrations were 55.0 %, 75.2 % and 77.8 %, respectively. Chloroquine (10 mg/kg of mouse, positive control) had a suppressive activity of 100 %.

Animals↗

Biological and pharmacological activities and further constituents of Hyptis verticillata.

Several extracts of Hyptis verticillata and isolated compounds were evaluated for their anti-inflammatory, antibacterial, antisecretory, and cytotoxic properties. The aerial parts yielded (R)-5-hydroxypyrrolidin-2-one and essential oil with the main components alpha-pinene, beta-pinene, and thymol. Spectroscopic methods (UV, IR, 1H-NMR, 13C-NMR, mass, CD) fully characterized (R)-5-hydroxypyrrolidin-2-one, as it was isolated by a bioassay guided fractionation. The essential oil, (R)-5-hydroxypyrrolidin-2-one, as well as the previously isolated rosmarinic acid and dehydropodophyllotoxine contributed to the antibacterial effects of H. verticillata. Furthermore, rosmarinic acid showed significant capillary stabilizing effects. Sideritoflavone inhibited prostaglandin synthase to a significant extent and had antisecretory effects comparable to those of NPPB. The cytotoxicity of the aqueous extract, as demonstrated using KB and HT 29 cell lines, may be of toxicological relevance in cases of internal application.

Animals↗

Effect of altering the type of enamel conditioner on the shear bond strength of a resin-reinforced glass ionomer adhesive.

The purpose of this study was to determine the effects of changing the type of enamel conditioner on the shear bond strength of a resin-reinforced glass ionomer within half an hour after bonding the bracket to the tooth. Freshly extracted human molars were collected and stored in a solution of 0.1% (weight/volume) thymol. The teeth were cleaned and polished. The teeth were randomly separated into 4 groups according to the enamel conditioner/etchant and adhesive used: group I, teeth were conditioned with 10% polyacrylic acid and brackets were bonded with a resin-reinforced glass ionomer adhesive; group II, teeth were conditioned with 20% polyacrylic acid and brackets were bonded with a resin-reinforced glass ionomer adhesive; group III, teeth were etched with 37% phosphoric acid and the brackets were bonded with a resin-reinforced glass ionomer adhesive; group IV, teeth were etched with 37% phosphoric acid and the brackets were bonded with a composite adhesive. The results of the analysis of variance comparing the 4 experimental groups (F = 24.87) indicated the presence of significant differences between the groups (P =.0001). In general, the shear bond strengths were significantly greater in the 2 groups etched with 37% phosphoric acid. This was true for both the resin-reinforced glass ionomer (X = 6.1 +/- 2.7 MPa) and the composite (X = 5.2 +/- 2.9 MPa) adhesives. On the other hand, the shear bond strengths were significantly lower in the two groups conditioned with polyacrylic acid. The bond strength of the resin-reinforced glass ionomer adhesive conditioned with 10% polyacrylic acid (X = 0.4 +/- 1.0 MPa) was significantly lower than the group conditioned with 20% polyacrylic acid (&xmacr; = 3.3 +/- 2.6 MPa). The present findings indicated that the bond strength of the resin-reinforced glass ionomer adhesive can be significantly increased in the initial half hour after bonding if the enamel is etched with 37% phosphoric acid instead of being conditioned with either 10% or 20% polyacrylic acid. The clinician needs to take these properties into consideration when ligating the initial archwires.

Acid Etching, Dental↗

Effect of changing enamel conditioner concentration on the shear bond strength of a resin-modified glass ionomer adhesive.

The purpose of this study was to determine the effects on shear bond strength of changing the concentration of the enamel conditioner used with resin-reinforced glass ionomer. Shear bond strength was measured within 30 minutes after bonding. Forty-six freshly extracted human molars were collected and stored in a solution of 0. 1% (weight/volume) thymol. The teeth were cleaned and polished, then randomly separated into 2 groups. In group I, enamel was conditioned with a 10% polyacrylic acid solution before bonding. In group II, the enamel was conditioned with a 20% polyacrylic acid solution. The results of t test comparisons of the 2 experimental groups (t = 4.9) indicate significant differences (P =.001). Shear bond strength was significantly greater in the group conditioned with the 20% polyacrylic acid concentration (X = 3.3 +/- 2.6 MPa) than in the group conditioned with the 10% concentration (X = 0.4 +/- 1.0 MPa). The present findings indicated that the lower initial shear bond strength of resin-reinforced glass ionomer adhesive can be improved more than 8-fold when the concentration of the polyacrylic acid enamel conditioner is increased from 10% to 20%. The clinician needs to take these properties into consideration when ligating the initial archwires.

Acid Etching, Dental↗

Effect of time on the shear bond strength of cyanoacrylate and composite orthodontic adhesives.

The purpose of this study was to compare the effects of time on the shear bond strength of a cyanoacrylate system and a composite adhesive system (1) within half an hour after bonding the bracket to the tooth, and (2) after 24 hours from the time of bonding when the adhesive had achieved most of its bond strength. Eighty freshly extracted molars were collected and stored in a solution of 0.1% (weight/volume) thymol. The teeth were cleansed, polished, and randomly separated into 4 groups: (I), cyanoacrylate adhesive debonded within one-half hour from initial bonding; (II), cyanoacrylate adhesive debonded after 24 hours immersion in deionized water at 37 degrees C; (III), composite adhesive debonded within one-half hour from initial bonding; (IV), composite adhesive debonded after 24 hours immersion in deionized water at 37 degrees C. The results of the analysis of variance comparing the 4 experimental groups (F = 12.68) indicated significant differences between them (P =.0001). The composite adhesive at 24 hours had significantly greater shear bond strength than did the other 3 groups. In general, the shear bond strengths were greater in the 2 groups debonded after 24 hours. This was true for both the cyanoacrylate ((-)x 7.1 +/- 3.3 megaPascals [MPa]) and the composite ((-)x 10.4 +/- 2.8 MPa) adhesives. On the other hand, the shear bond strength was significantly lower in the 2 groups debonded within one-half hour of their initial bonding. The bond strength of the cyanoacrylate adhesive ((-)x 5.8 +/- 2.4 MPa) was not significantly different from that of the composite ((-)x 5.2 +/- 2.9 MPa) adhesive. Our findings indicated that the cyanoacrylate and the composite adhesives tested have clinically adequate shear bond strengths at half an hour and at 24 hours after initial bonding. The clinician needs to consider the properties of each adhesive: eg, the need to use a curing light and the ability to have more working time with the composite adhesive versus no light but only a 5-second working time before the cyanoacrylate adhesive starts to set.

Analysis of Variance↗

Singlet oxygen and peroxyl radicals regulate carotenoid biosynthesis in Phaffia rhodozyma.

Carotenoids have recently received considerable interest because of their potential in delaying or preventing degenerative diseases such as arteriosclerosis, cancer, and aging. In this study we show that the active oxygen species singlet oxygen (1O2) and peroxyl radicals differently affect carotenoid composition and biosynthesis in the yeast Phaffia rhodozyma. Photochemical generation of 1O2 with rose bengal or alpha-terthienyl induced carotenoid accumulation. In contrast, peroxyl radicals derived from t-butylhydroperoxide (tBOOH) or H2O2 decreased the content of astaxanthin and increased beta-carotene by approximately 4-fold, suggesting end product feedback regulation by astaxanthin or inhibition of biosynthetic enzymes. 14C labeling of carotenoids during oxidative stress supported the possibility of end product regulation. Carotenoids were bleached by 8 mM tBOOH within 6 h when carotenogenesis was inhibited by thymol. When treated with peroxides, a previously unreported pigment in P. rhodozyma was formed. The carotenoid had a mass of 580 Da and a molecular formula of C40H52O3. Chemical derivatizations combined with mass and absorbance spectroscopy tentatively identified the carotenoid as dehydroflexixanthin (3,1'-dihydroxy-2,3,3',4'-tetradehydro-1',2'-dihydro-beta,psi-caro tene-4-one). This study provides the first report of induction of astaxanthin biosynthesis by 1O2, probable feedback control by astaxanthin, and the oxidative degradation of astaxanthin to novel pigments in P. rhodozyma.

Carotenoids↗

Development of a paw paw herbal shampoo for the removal of head lice.

The development and clinical testing of an herbal lice removal shampoo, containing a standardized extract of paw paw, thymol, and tea tree oil, are described. All of these ingredients were selected for their ability to deplete adenosine triphosphate (ATP) levels and, thus, prevent ATP-dependent pesticide resistance. Optimum concentrations of the ingredients, treatment times, and dosing schedules were established through in vitro tests with head lice. In addition to pilot studies involving 21 participants, a final clinical trial, using the optimum shampoo formulation in 16 participants, demonstrated 100% effectiveness in removing head lice and nits.

Adult↗

Respiratory toxicity of fabric softener emissions.

To determine whether there is any biological basis for complaints that fabric softener emissions can cause acute adverse effects in certain individuals, screening tests were performed in which groups of mice were exposed to the emissions of 5 commercial fabric softener products (antistatic pads used in laundry dryers) for 90 min. Pneumotachographs and a computerized version of ASTM test method E-981 were used to measure acute changes in several respiratory cycle parameters, especially the pause after inspiration, the pause after expiration, and the midexpiratory airflow velocity. From these changes, sensory irritation (SI), pulmonary irritation (PI), and airflow limitation (AFL) of differing intensities were measured with each of the five brands tested. At the peak effect, SI ranged from 21 to 58% of the breaths, PI ranged from 4 to 23% of the breaths, and AFL ranged from 6 to 32% of the breaths. After three exposures, histopathology revealed mild inflammation of interalveolar septae of the lungs. Gas chromatography/ mass spectroscopy (GC/MS) analysis of the emissions of one pad identified several known irritants (isopropylbenzene, styrene, trimethylbenzene, phenol, and thymol). Laundry that had been dried with one the fabric softener pads emitted sufficient chemicals to elicit SI in 49% of breaths at the peak effect Placing one fabric softener pad in a small room overnight resulted in an atmosphere that caused marked SI (61% of breaths). These results demonstrate that some commercial fabric softeners emit mixtures of chemicals that can cause SI, PI, and reduce midexpiratory airflow velocity in normal mice. The results provide a toxicological basis to explain some of the human complaints of adverse reactions to fabric softener emissions.

Air Pollutants↗

A method for quantitative analysis of flavor-tainting alkylphenols and aromatic thiols in fish.

Simultaneous steam distillation-extraction (SDE) of fortified rainbow trout tissue resulted in greater than 95% recovery of 2-isopropylphenol, 3-isopropylphenol, 4-isopropylphenol, 2,4-diisopropylphenol, 2,5-diisopropylphenol, 3,5-diisopropylphenol, carvacrol and thymol. Lower recoveries were obtained for 2,6-diisopropylphenol (81%), thiophenol (55%), and thiocresol (85%). Analysis of concentrated extracts by gas chromatography/mass spectrometry operated in the selected ion monitoring mode allowed quantitative detection of these compounds down to 0.5 ppb based on 20 g of initial sample.

Animals↗

Comprehensive study on vitamin C equivalent antioxidant capacity (VCEAC) of various polyphenolics in scavenging a free radical and its structural relationship.

Antioxidant capacity for a wide range of natural or synthetic polyphenolics was comprehensively evaluated by vitamin C equivalent antioxidant capacity (VCEAC) assay using free blue/green ABTS radicals. The polyphenolics tested are grouped into the following categories: vitamins (beta-carotene, alpha-tocopherol, vitamin A, and vitamin C), phenolic acids (benzoic acid, phenylacetic acid, cinnamic acid, and their derivatives), flavonoids (anthocyanidin, flavanol, chalcone, flavanone, flavone, flavonol, isoflavone, and their derivatives), synthetic food additives (BHA, BHT, TBHQ, and PG), and other miscellaneous polyphenolics (ellagic acid, sesamol, eugenol, thymol, etc.). A positive linear relationship between VCEAC and the number of free OH groups around the flavonoid framework was found, whereas, for phenolic acids, the linear relationship was not as good as with the flavonoid aglycones. Groups of chemicals having comparable structures generally showed similar trends. Polyphenolics commonly showed a higher VCEAC compared to monophenolics. Compounds like gallic acid with 3 vicinal hydroxy substitutions on the aromatic ring in phenolic acids or like epigallocatechin with 3 vicinal hydroxy substitutions on the B ring in flavonoids showed the highest antioxidant capcity among the groups. In the flavonoids, 2 characteristic chemical structures were very important, the catechol moiety in the B ring and the 3-OH functional group in a chroman ring. Glycosylated flavonoids showed less potent antioxidant capacity than their aglycone alone. Synthetic antioxidant food additives (BHA, TBHQ, and BHT) conventionally used in the food industry were less effective antioxidants than ascorbic acid. Other naturally occurring polyphenolics tested followed the expected general trends of phenolic acids and flavonoids.

Antioxidants↗

GC-MS analysis and bioactivity testing of the volatile oil from the leaves of the toothbrush tree Salvadora persica L.

Upon GC-MS analysis of the volatile oil extracted from Salvadora persica L. leaves, we were able to identify benzyl nitrile, eugenol, thymol, isothymol, eucalyptol, isoterpinolene, and beta-caryophyllene. Toxicity of the aroma was evaluated using brine shrimp lethality test which gave an LC50 > 1,000 ppm. Using Disc Diffusion Test, it was found that the extract of the leaves has a considerable antibacterial effect on several different oral aerobic bacteria with comparable results to known antibiotics. The extract can be used effectively as a natural tool for teeth cleaning and as a natural analgesic for the disturbing toothache.

Analgesics↗