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Antibacterial and antioxidant activities of essential oils isolated from Thymbra capitata L. (Cav.) andOriganum vulgare L.

Antilisterial activities of Thymbra capitata and Origanum vulgare essential oils were tested against 41 strains of Listeria monocytogenes. The oil of T. capitata was mainly constituted by one component, carvacrol (79%), whereas for O. vulgare three components constituted 70% of the oil, namely, thymol (33%), gamma-terpinene (26%), and p-cymene (11%). T. capitata essential oil had a significantly higher antilisterial activity in comparison to O. vulgare oil and chloramphenicol. No significant differences in L. monocytogenes susceptibilities to the essential oils tested were registered. The minimum inhibitory concentration values of T. capitata essential oil and of carvacrol were quite similar, ranging between 0.05 and 0.2 microL/mL. Antioxidant activity was also tested, the essential oil of T. capitata showing significantly higher antioxidant activity than that of O. vulgare. Use of T. capitata and O. vulgare essential oils can constitute a powerful tool in the control of L. monocytogenes in food and other industries.

Anti-Bacterial Agents↗

Antioxidant and biocidal activities of Carum nigrum (seed) essential oil, oleoresin, and their selected components.

In the present study, chemical constituents of the essential oil and oleoresin of the seed from Carum nigrum obtained by hydrodistillation and Soxhlet extraction using acetone, respectively, have been studied by GC and GC-MS techniques. The major component was dillapiole (29.9%) followed by germacrene B (21.4%), beta-caryophyllene (7.8%), beta-selinene (7.1%), and nothoapiole (5.8%) along with many other components in minor amounts. Seventeen components were identified in the oleoresin (Table 2) with dillapiole as a major component (30.7%). It also contains thymol (19.1%), nothoapiole (15.2.3%), and gamma-elemene (8.0%). The antioxidant activity of both the essential oil and oleoresin was evaluated in mustard oil by monitoring peroxide, thiobarbituric acid, and total carbonyl and p-anisidine values of the oil substrate. The results showed that both the essential oil and oleoresin were able to reduce the oxidation rate of the mustard oil in the accelerated condition at 60 degrees C in comparison with synthetic antioxidants such as butylated hydroxyanisole and butylated hydroxytoluene at 0.02%. In addition, individual antioxidant assays such as linoleic acid assay, DPPH scavenging activity, reducing power, hydroxyl radical scavenging, and chelating effects have been used. The C. nigrum seed essential oil exhibited complete inhibition against Bacillus cereus and Pseudomonas aeruginosa at 2000 and 3000 ppm, respectively, by agar well diffusion method. Antifungal activity was determined against a panel of foodborne fungi such as Aspergillus niger, Penicillium purpurogenum, Penicillium madriti, Acrophialophora fusispora, Penicillium viridicatum, and Aspergillus flavus. The fruit essential oil showed 100% mycelial zone inhibition against P. purpurogenum and A. fusispora at 3000 ppm in the poison food method. Hence, both oil and oleoresin could be used as an additive in food and pharmaceutical preparations after screening.

Allyl Compounds↗

Effect of thyme essential oils (Thymus hyemalis and Thymus zygis) and monensin on in vitro ruminal degradation and volatile fatty acid production.

The effect of the essential oils of thyme on the in vitro ruminal degradability of a barley seed/alfalfa hay substrate was studied. Two essential oils were used, one from Thymus hyemalis (TH), rich in carvacrol, and the other from Thymus zygis (TZ), rich in thymol. Four experimental treatments of in vitro degradability, using the Daisy II(200/220) incubator, were conducted including a negative control (CO), a positive control at 7.5 microg/mL of monensin (MO), and two treatments with essential oils (TH or TZ) at 1.35 microL/mL. The material was incubated at 39.5 degrees C for various lengths of time. At each time, the disappearance of dry matter, crude protein, and neutral detergent fiber was measured. Volatile fatty acids (VFAs) were determined after 48 h of incubation. CO and MO provided (p < 0.01) higher values of potential degradability (a + b) of DM than the TH and TZ treatments (72.6 and 70.8 vs 53.2 and 48.2%, respectively). Also, crude protein degradability was lowest in the essential oil treatments. The CO treatment showed the highest potential degradability of NDF. The values of VFA production obtained (p < 0.001) with CO and MO treatments were higher than those obtained with TH and TZ treatments (21.0 and 19.1 vs 11.2 and 10.1 mM). The essential oils decreased the molar proportion of propionate, increasing the acetate/propionate ratio. In conclusion, the effects of essential oils at assayed doses would not be nutritionally beneficial to the ruminal energetic metabolism.

Animals↗

Floral Origin Markers of Chestnut and Lime Tree Honeys.

The apiculture industry is more and more interested in finding typical markers to authenticate floral origin of honeys. With this aim, some reliable volatile compounds were proposed to identify origin of lime tree and chestnut samples. A dichloromethane extraction followed by a Likens-Nickerson simultaneous steam distillation/solvent extraction led to representative honey extracts. About 400 volatile compounds were separated by gas chromatography, but only a few authenticated the floral origin of honeys. Chestnut honeys are distinguishable from other origins by high concentrations of acetophenone, 1-phenylethanol (>88 ppb), and 2-aminoacetophenone (>154 ppb). Lime tree honeys are characterized by enhanced amounts of shikimate pathway derivatives (ethylmethylphenol isomer (>31 ppb), 4-tert-butylphenol, estragole (>51 ppb), and p-methylacetophenone but also by high concentrations of monoterpene-derived compounds (menthol, thymol, 8-p-menthene-1,2-diol, and carvacrol (>76 ppb)) and methyl(1-methylethenyl)benzene.

Journal Article↗

The main citral-geraniol and carvacrol chemotypes of the essential oil of thymus pulegioides L. growing wild in vilnius district (Lithuania).

Thymus pulegioides L. with lemon and carvacrol odor form the major part of plants growing wild in all 10 investigated localities during 1995-1997. The main components of the citral-geraniol chemotype of lemon-scented essential oil are the following (%): geraniol (14.9-30. 8), geranial (trans-citral, 9.7-19.7), beta-caryophylene (6.0-11.4), nerol (4.1-11.8), and neral (cis-citral, 0.1-9.5). The essential oil of carvacrol chemotypes contain more compounds that are characteristic of the thyme genus (%): carvacrol (16.0-22.2), beta-bisabolene (11.1-20.2), beta-caryophyllene (11.1-19.1), gamma-terpinene (5.8-16.2), p-cimene (5.5-10.4), thymol (3.3-9.8), and carvacrol methyl ether (5.6-8.6). The correlation between the odor and composition of the essential oil will help the users of wild thyme to choose the necessary chemotype for their purposes.

Acyclic Monoterpenes↗

Sesquiterpenes and monoterpenes from the bark of Inula macrophylla.

Eleven new sesquiterpenes (1--11) and two thymol derivatives (12, 13), along with 12 known sesquiterpenes and monoterpenes, were isolated from the bark of Inula macrophylla. Their structures were determined on the basis of spectral evidence (especially by HREIMS and 2D NMR) as well as chemical transformations. The structure of macrophyllic acid A (1) was confirmed by X-ray analysis, and the absolute configuration of 1 was determined on the basis of the appropriate chemical conversions and the application of a modified Mosher's method.

Crystallography, X-Ray↗

Isolation and structure elucidation of radical scavengers from Thymus vulgaris leaves.

2,2-Diphenyl-1-picrylhydrazyl radical (DPPH*) scavenging activity-guided fractionation of a leaf extract of Thymus vulgaris led to the isolation of the radical scavengers rosmarinic acid 1, eriodictyol, taxifolin, luteolin 7-glucuronide, p-cymene 2,3-diol, p-cymene 2,3-diol 6-6'-dimer, carvacrol, thymol, and a new compound, 2. The fractionation was considerably facilitated by using an on-line HPLC detector for radical scavenging activity. In this detector activity is monitored as the disappearance of the color of a postcolumn added stable radical after reacting with radical scavengers in a reaction coil. Compound 2, which consists of rosmarinic and caffeic acid moieties linked via a C-3'-C-8' ' ether bridge, was mainly elucidated by various NMR techniques and CD. Phenylpropanoid trimer 2 was a weaker and stronger radical scavenger than rosmarinic acid 1 in off-line TEAC and DPPH* assays, respectively.

Biphenyl Compounds↗

Bioactivity of the essential oil of Bupleurum fruticosum.

The essential oil of Bupleurum fruticosum was investigated qualitatively and quantitatively together with the anti-inflammatory activity of the whole essential oil and its major components. In addition, antispasmodic activity was determined in rat uterus preparations using acetylcholine and oxytocin as agonists. The anti-inflammatory activity shown by the essential oil can be attributed in part to the two major components, alpha-pinene and beta-pinene, although the presence of thymol and carvacrol, minor components capable of potentiating the action of these hydrocarbons, was also confirmed.

Acetylcholine↗

Molecular structure influence in the recognition of phthaleins by the electrogenic organic anion carrier at the sinusoidal plasma membrane level in the liver.

The hepatocytic uptake of cholephilic organic anions occurs by carrier-mediated mechanisms. Electrogenic and electroneutral transport systems have been described. The aim of this study was to determine the dissociation constant (Kd) of the Electrogenic Carrier System(s) (ECS) for tetrabromosulfophthalein (BSP), dibromosulfophthalein (DBSP), tetrabromophthalein (TBP), tetrabromosulfonephthalein (TBS) and thymol blue (TB). Kd (uM) values for ECS-organic anion complexes were: ECS-BSP = 3.61 +/- 0.18; ECS-DBSP = 11.61 +/- 1.32; ECS-TBP = 0.51 +/- 0.08; ECS-TBS = 1.31 +/- 0.25; ECS-TB = 9.44 +/- 1.80. From these data, it is possible to conclude that molecular characteristics of the organic anions are important factors in determining the dissociation constant for the electrogenic hepatic carrier(s). In this sense, the addition of two sulphonic groups on the phenolic ring; the presence of a sulphonic on the benzenic ring and the absence of two or four bromines on the molecule confers a lower affinity for ECS.

Animals↗

Mechanism of transport enhancement of LHRH through porcine epidermis by terpenes and iontophoresis: permeability and lipid extraction studies.

PURPOSE: The purpose of this study was to investigate the effect of 5% terpenes (i.e., limonene, carvone, thymol, and cineole)/ethanol (EtOH) and iontophoresis on the in vitro permeability of luteinizing hormone releasing hormone (LHRH) through the porcine epidermis and biophysical changes in the stratum comeum (SC) lipids by fourier transform infrared (FT-IR) spectroscopy. Methods. The porcine epidermis was pretreated with enhancer for 2 h. The permeability measurement system included Franz diffusion cells, Ag/AgCl electrodes, and SCEPTOR iontophoretic power source. FT-IR spectroscopy was performed to assess the possible contribution of lipid extraction to the transport enhancement of LHRH. RESULTS: Terpenes in combination with EtOH significantly (p < 0.05) increased the flux of LHRH in comparison with the control (epidermis which was not enhancer treated). Iontophoresis further enhanced (p < 0.05) the flux of LHRH through terpenes/EtOH treated epidermis in comparison with their passive permeability. Reversibility studies showed that the post-recovery passive flux of LHRH through 5% limonene in EtOH/iontophoresis treated epidermis was significantly (p < 0.05) decreased but did not significantly recover to the baseline flux (i.e., flux through control epidermis). The SC treated with terpenes/ EtOH showed a decrease in peak heights and areas for both asymmetric and symmetric C-H stretching absorbances in comparison to untreated SC. A greater percent decrease in peak heights and areas was obtained by limonene/EtOH. However, treatment of the SC with terpenes/EtOH followed by iontophoresis did not further decrease the percentage of peak height and area over and above terpene/EtOH suggesting that iontophoresis alone does not cause SC lipid extraction. CONCLUSIONS: Terpenes/EtOH increased LHRH permeability by enhancing the extraction of the SC lipids. Iontophoresis synergistically enhanced the permeability of LHRH through terpenes/EtOH treated epidermis. Thus, terpenes can be used as chemical enhancers in combination with iontophoresis to enhance the transdermal delivery of peptides such as LHRH.

Administration, Cutaneous↗

Antifungal activities of origanum oil against Candida albicans.

The antimicrobial properties of volatile aromatic oils from medicinal as well as other edible plants has been recognized since antiquity. Origanum oil, which is used as a food flavoring agent, possesses a broad spectrum of in vitro antimicrobial activities attributed to the high content of phenolic derivatives such as carvacrol and thymol. In the present study, antifungal properties of origanum oil were examined both in vitro and in vivo. Using Candida albicans in broth cultures and a micro dilution method, comparative efficacy of origanum oil, carvacrol, nystatin and amphotericin B were examined in vitro. Origanum oil at 0.25 mg/ml was found to completely inhibit the growth of C. albicans in culture. Growth inhibitions of 75% and >50% were observed at 0.125 mg/ml and 0.0625 mg/ml level, respectively. In addition, both the germination and the mycelial growth of C. albicans were found to be inhibited by origanum oil and carvacrol in a dose-dependent manner. Furthermore, the therapeutic efficacy of origanum oil was examined in an experimental murine systemic candidiasis model. Groups of mice (n = 6) infected with C. albicans (5 x LD50) were fed varying amounts of origanum oil in a final vol. of 0.1 ml of olive oil (vehicle). The daily administration of 8.6 mg of origanum oil in 100 microl of olive oil/kg body weight for 30 days resulted in 80% survivability, with no renal burden of C. albicans as opposed to the group of mice fed olive oil alone, who died within 10 days. Similar results were obtained with carvacrol. However, mice fed origanum oil exhibited cosmetically better clinical appearance compared to those cured with carvacrol. The results from our study encourage examination of the efficacy of origanum oil in other forms of systemic and superficial fungal infections and exploration of its broad spectrum effect against other pathogenic manifestations including malignancy.

Animals↗

Copper and dyes enhance laccase production in gamma-proteobacterium JB.

Laccase production in gamma-proteobacterium JB was enhanced 13-fold by adding 0.1 mM CuSO(4) 24 h after the onset of growth. Ethidium bromide (2.5 microM), Malachite Green, Phenol Red and Thymol Blue (10 microM each) enhanced laccase production 17-, 19-, 4- and 2-fold, respectively. Among the fourteen aromatic/organic compounds tried, p-aminobenzoic acid and an industrial effluent, from where the organism was isolated, showed 1.2- and 1.26-fold increases in production.

Coloring Agents↗

A calcium-binding protein with similarity to serum albumin localized to the ER-Golgi network and cell walls of spinach (Spinacia oleracea).

Using polyclonal antibodies raised against human serum albumin (HSA), a 70-kDa microsomal protein with an isoelectric point of approximately 6.5 was detected in spinach (Spinacia oleracea L.). The protein was purified by selective ammonium sulfate precipitation and anion exchange HPLC. The protein shared 100% identity with the first 15 amino acids at the NH2 terminus of HSA, including the X-X-H amino acid region, which was identified in HSA as being responsible for binding of copper, zinc, indole derivatives and calcium. Blue staining of the protein with the cationic carbocyanine dye 'Stains-all' and 45Ca overlay following SDS-PAGE also suggest that the 70-kDa plant protein binds calcium. The protein reacted positively with carbohydrate specific thymol stain, and the carbohydrates associated with the protein were identified by gas chromatography-mass spectrometry (GC-MS) as galactose and galacturonic acid. The 70-kDa plant protein was present in the detergent-poor phase following Triton X-114 extraction of the microsomal proteins. Cell fractionation using continuous sucrose gradients showed that the protein is present in membrane fractions with high activity of endoplasmic reticulum (ER) and Golgi marker enzymes. Using nitrocellulose tissue prints probed with anti-HSA antibodies, we demonstrated that the protein is present in the apoplastic space of petioles, suggesting that the protein is secreted to the apoplast of cortex cells in plants. Localization and binding properties suggest that the plant protein identified in the present study may participate in secretion processes, possibly involved with the transport of precursors required for cell-wall synthesis.

Journal Article↗

Two-parameter determination in vinegar by a flow injection-pervaporation system.

A flow injection method (FI) for the sequential determination of ethanol and acetic acid in vinegar is reported. The determination of ethanol is based on the oxidation of the pervaporated ethanol by K2Cr2O7. The acetic acid is determined by an acid-base reaction with Thymol Blue as the indicator. Both reactions are monitored photometrically at 600 nm using a single detector. Optimisation studies and assessment of the sequential Fl method are also reported. The linear determination range is 0-12% v/v for ethanol and 0-10% (grams of acetic acid in 100 ml) for acetic acid. The sample throughput of the sequential manifold is seven per hour. The new method was applied to vinegar samples and the results obtained were in excellent agreement with those from reference methods used in Spain.

Acetic Acid↗

The effect of repeated bonding on the shear bond strength of a composite resin orthodontic adhesive.

One of the problems clinicians face during treatment is bracket failure. This is usually the result either of the patient's accidentally applying inappropriate forces to the bracket or of a poor bonding technique. As a result, a significant number of teeth have to be rebonded in a busy orthodontic practice. The purpose of this study was to evaluate the effect of repeated bonding on the shear bond strength of orthodontic brackets. Fifteen freshly extracted human molars were collected and stored in a solution of 0.1% (wt/vol) thymol. The teeth were cleaned, polished, and etched with a 37% phosphoric acid gel. The brackets were bonded with the adhesive and light cured for 20 seconds. The teeth were sequentially bonded and debonded 3 times with the same composite orthodontic adhesive. At each time, all 15 teeth were debonded within a half hour after bonding to simulate the clinical condition at which a newly bonded bracket is attached to the arch wire. The results of the analysis of variance comparing the shear bond strength at the 3 debonding attempts indicated the presence of no significant differences among the 3 groups (P = .104). However, when the overall change in shear bond strength within each tooth was evaluated between debonding sequences 1 and 3, 10 teeth had a significant (P = .001) decrease (mean +/- SD, -4.6+/-2.5 MPa) in bond strength, whereas 5 teeth had a significant (P = .02) increase (mean +/- SD, 2.8+/-1.6 MPa). The present findings indicated that in general, the highest values for shear bond strength were obtained after the initial bonding. Rebonded teeth have significantly lower and inconsistent shear bond strength; ie, bond strength may further decrease or increase after the second debonding, and the changes in bond strength may be related to the changes in the morphologic characteristics of the etched enamel surface as a result of the presence of adhesive remnants.

Analysis of Variance↗

Effects of alternating and direct electrical current application on the odontoblastic layer in human teeth: an in vitro study.

AIM: The aim of this study was to investigate the influence of a low intensity alternating current on the odontoblasts and odontoblast layer and compare this with the effects of a direct current. METHODOLOGY: Teeth extracted for orthodontic reasons were immersed in physiological saline stabilized with thymol crystals. Within 1 h of extraction, an alternating or direct current was applied on the crown in the direction of the apex of the tooth for 120-360 s. The current doses were 12, 30, 60, 600, 1800, 3600, 7200, 24,000 and 144,000 microC. The teeth were fixed in Bouin or Baker fluids, the pulps removed, dehydrated and immersed in paraffin, then sectioned, stained with haematoxylin and eosin, and studied under a light microscope. RESULTS: Neither direct nor alternating current, similar to that applied in electrical caries diagnosis caused histological changes in the odontoblasts. CONCLUSIONS: There was no difference between direct and low intensity alternating current in the response of the odontoblast.

Adolescent↗

Accuracy of a new apex locator: an in vitro study.

AIM: The purpose of this study was to test in an in vitro model the accuracy of a Bingo 1020 electronic apex locator, to compare the results to those of a well known apex locator, Root ZX, as well as to those of the radiographic method of tooth length determination. METHODOLOGY: A total of 120 extracted teeth, preserved in Thymol solution and kept refrigerated, was used for the study. The experiment was performed on single-rooted teeth and on one-root canal, chosen randomly, in multirooted teeth. The teeth were randomly divided into 12 groups of 10 teeth each. After access preparation, the actual length (AL) was measured. The teeth were embedded in an alginate model specially developed for testing apex locators. Electronic tooth length measurements (EL) were carried out prior to root canal preparation using the two electronic apex locators (EAL) - Root ZX and Bingo 1020; three measurements were taken and an average computed. After the third measurement, the file was left in the root canal and a periapical radiograph was taken. The radiographic length (RL) was recorded by measuring the file length from the coronal reference point to the tip of the file. Each root canal was then prepared to a no. 40 K-file diameter using a standardized technique; saline was used for irrigation. Upon completion of the root canal preparation, EL measurements were taken by each EAL in dry conditions and with different irrigation solutions. Each measurement was repeated three times. The RL was recorded according to the last EL measurement. Results were subjected to statistical analysis. RESULTS: In all parameters tested, a significant statistical difference was found between Bingo 1020 and the Root ZX. Measurements obtained using the Bingo 1020 were consistently closer to the AL (0.08 mm) than those obtained using the Root ZX. Both EALs measured the tooth length with great accuracy and a positive correlation of 0.76 (P = 0.00) existed between the two devices. No significant difference was found between the two apex locators when measurements were taken with the different irrigants (P = 0.34) and the content of the root canal did not affect the accuracy of the measurements. Lengths obtained by calculations from the radiographs were longer than the AL as well as the length obtained by both EALs (P = 0.00). CONCLUSIONS: The Bingo 1020 proved to be as reliable as Root ZX and was user friendly. Under the experimental conditions, electronic measurements were more reliable than radiographs in the process of root length determination.

Analysis of Variance↗

The influence of three Erbium:YAG laser energies on the in vitro microleakage of Class V compomer resin restorations.

OBJECTIVES: To investigate the influence of three Erbium:YAG pulse energies (200 mJ, 240 mJ, and 300 mJ with a 100 mJ dentine finish) on the microleakage of Class V compomer restorations (Compoglass). DESIGN: In vitro study. SAMPLE AND METHODS: Sixty-one extracted human premolar teeth were randomly allocated to three groups (according to pulse energy). Each tooth hosted one test cavity prepared with one of the three Er:YAG pulse energies using a pulse repetition rate of 5 Hz and a pulse duration of 250 microseconds, and one control cavity prepared with a conventional diamond bur in a high-speed hand piece. Both cavities were placed at the cervical margin of the tooth and were restored and finished according to the manufacturer's instructions. Each tooth was then stored in 0.12% thymol solution at 37 degrees C for three months, before being thermocycled through water baths between 5 degrees C and 55 degrees C for 240 cycles. Microleakage was assessed using a methylene blue dye penetration technique and was quantified using a score 0 (none) to 4 (to and into the axial wall). The data was analysed using the Kruskal-Wallis and Mann-Whitney U-tests. RESULTS: Leakage was seen in all groups at both the dentine and enamel margins. There were no statistically significant differences in leakage (P < 0.05) at either the enamel or dentine margins, except for the 240 mJ lased enamel margins. This group performed significantly better than the enamel margins prepared with 200 mJ (P = 0.03) and the 300/100 mJ (P = 0.01) laser energies as well as the conventionally prepared enamel margins (P < 0.001). CONCLUSION: Cavity preparation with this Erbium:YAG laser did not influence the microleakage of Compoglass restorations adversely. Different pulse energies were required for optimum cavity sealing at the enamel and dentine margins and for different materials.

Bicuspid↗