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In vitro antimicrobial and antioxidant activities of the essential oils and various extracts of Thymus eigii M. Zohary et P.H. Davis.

This study was designed to examine the in vitro antimicrobial and antioxidant activities of the essential oil and various extracts obtained from aerial parts of Thymus eigii. The essential oil was particularly found to possess stronger antimicrobial activity, whereas other nonpolar extracts and subfractions showed moderate activity and polar extracts remained almost inactive. GC-MS analysis of the oil resulted in the identification of 39 compounds, representing 93.7% of the oil; thymol (30.6%), carvacrol (26.1%), and p-cymene (13.0%) were the main components. The samples were also subjected to a screening for their possible antioxidant activity by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and beta-carotene-linoleic acid assays. In the former case, the polar subfraction of the methanol extract was found to be superior to all extracts tested, only 16.8 microg/mL of which provided 50% inhibition, whereas all extracts, particularly the polar ones, seem to inhibit the oxidation of linoleic acid in the latter case. These data were further supported by total phenolics analysis, indicating that the antioxidative potential of the extracts was closely related to their phenolic constituents.

Anti-Bacterial Agents↗

Microbiological and clinical effects of an antiseptic dental varnish after mechanical periodontal therapy.

The aim of this study was to explore the microbiological and clinical effects of an antiseptic dental varnish when applied to periodontally diseased teeth after mechanical therapy. 20 subjects participated in this placebo controlled, double blind prospective longitudinal study. 2 experimental sites with a pocket probing depth > or =5 mm were chosen in each subject. The control varnish, consisting of ethanol, ethylacetate and polyvinylbutyral, was applied to one of the selected teeth and the test varnish, containing 1% chlorhexidine and 1% thymol in addition, was applied to the other one. Clinical parameters were assessed, and microbiological samples were obtained from the two study sites at baseline (6-10 weeks after completion of conventional periodontal therapy), and 2, 4 and 12 weeks thereafter. The mean PLI at baseline was very low and, therefore, only a minimal potential for a further improvement existed. During the 12-week observation period, the mean PLI increased significantly at sites treated with the placebo varnish, while no similar trend for an increase in PLI was detected in the test sites. The bleeding tendency seemed to remain unaffected by the application of the varnish. On the microbiological level, no relevant differences could be detected between placebo and test sites at baseline, or during the follow-up period. In conclusion, the application of a dental varnish with antimicrobial properties after mechanical periodontal therapy had little effect in subjects with good oral hygiene.

Adult↗

Effects of fluoride- and chlorhexidine-containing varnishes on plaque composition and on demineralization of dentinal grooves in situ.

The aim of the study was to compare the effects of Cervitec, containing 1% chlorhexidine (CHX) and 1% thymol, Fluor Protector, containing 0.1% fluoride, their 1:1 mixture, and a placebo varnish on the percentage of mutans streptococci and lactobacilli in plaque and on the underlying dentin demineralization, as assessed by microradiography. Bovine dentine discs, fitted with three parallel grooves, received one of the varnish treatments into the first groove and on the adjacent part of the dentin surface. Volunteers (n = 23) wore the discs fixed to their partial dentures for four consecutive 3 wk periods. Microbiological analysis of plaque accumulated in the grooves showed no difference between groups. Fluoride varnishes (Fluor Protector and mixed varnish) had a significantly larger inhibitory effect on mineral loss in the treated groove than Cervitec or placebo. All treatment varnishes had more pronounced effect in panelists (n = 14) with higher degree of demineralization (mineral loss in placebo group > or = 1,200 vol% x microm). In these panelists, CHX-containing varnishes showed an inhibitory effect on demineralization in all grooves, also in the two non-varnished grooves. As fluoride varnishes had the largest localized effect on demineralization, and CHX varnishes were showing a peripheral effect, a combined treatment could be the preferred method to obtain an optimal caries preventive effect in caries-prone individuals.

Aged↗

The effects of calcium on protein turnover in skeletal muscles of the rat.

Several experimental procedures were used to increase the intracellular concentration of Ca2+ and determine its effects on protein turnover in isolated extensor digitorum longus and soleus muscle. These methods included the use of ionophore A23187, caffeine, dibucaine, thymol and procaine, all agents known to induce the release of calcium by acting either on the sarcolemma and/or on the sarcoplasmic reticulum. Another approach involved varying the external concentration of Ca2+ in the media in which the muscles were incubated. The changes in muscle Ca2+ concentrations after exposure to the various calcium-releasing agents were in keeping with accepted modes of action of these agents on muscle membranes. The findings suggest that increasing the sarcoplasmic concentration of Ca2+ inhibits protein synthesis and enhances protein breakdown. These catabolic effects of Ca2+ are compared with the changes induced in muscle protein turnover after exposure to insulin or cyclic nucleotides, and in myopathic muscle and situations of work overload. Attention is also drawn to some of the difficulties involved in definitively implicating Ca2+ as a factor involved in the normal regulation of protein turnover.

Animals↗

Effect of a chlorhexidine/thymol-containing varnish on prostaglandin E2 levels in gingival crevicular fluid.

The aim was to study the effect of a chlorhexidine/thymol-containing varnish (Cervitec) on the levels of prostaglandin E2 (PGE2) in gingival crevicular fluid (GCF). The material consisted of 25 adolescents and young adults with fixed orthodontic appliances exhibiting gingival inflammation. Four buccal sites, adjacent to bands and brackets, were selected on each patient and randomly treated with either a varnish containing chlorhexidine diacetate (1% w/w) and thymol (1% w/w) or a placebo varnish without active ingredients. After baseline registration, the varnishes were applied twice within 3 d. Follow-up examinations were performed after 3, 8 and 30 d. The gingival inflammation was assessed by bleeding on probing, volume of GCF with a Periotron 8000 and PGE2 level in GCF by using a radioimmuno assay. Compared with baseline, a statistically significant reduction in the volume of GCF was recorded at the chlorhexidine/thymol treated sites in contrast to the placebo. The mean PGE2 levels were significantly reduced after the test varnish treatment compared with baseline and differed significantly from placebo after 8 d. The findings suggest that treatments with the antibacterial varnish result in reduced gingival inflammation and may thus be beneficial for patients with fixed orthodontic appliances.

Adolescent↗

Susceptibility testing: inoculum size dependency of inhibition using the Colworth MIC technique.

The minimum inhibitory concentration, MIC, is an accepted and well used criterion for measuring the susceptibility of organisms to inhibitors. Many factors influence the MIC value obtained, including temperature, inoculum size and type of organism. A modification of the method developed in this laboratory to obtain inhibition profiles of antimicrobials was used to examine the effect of inoculum size on the degree of inhibition observed with respect to inhibitor concentration. The data obtained enabled the production of an empirical model of inhibition, based on a Gompertz function, relating the level of growth observed to both the inoculum size and concentration of the inhibitor. The inoculum size dependencies of phenethyl alcohol, phenoxyethanol, p-chloro-m-cresol, trichloro-phenol, thymol and dodecyltrimethylammmonium bromide against Staphylococcus aureus were obtained.

Anti-Bacterial Agents↗

In-vitro antimicrobial activity and chemical composition of Sardinian Thymus essential oils.

Essential oils and their components are becoming increasingly popular as naturally occurring antimicrobial agents. In this work the chemical composition and the antimicrobial properties of Thymus essential oils and of their main components were determined. Three essential oils obtained from different species of Thymus growing wild in Sardinia and a commercial sample of Thymus capitatus oil were analysed. The essential oil components were identified by GC/MS analysis. The antimicrobial activity of the oils and components was determined against a panel of standard reference strains and multiple strains of food-derived spoilage and pathogenic bacteria, using a broth microdilution method. The GC/MS analysis showed that the major constituents of the oils were monoterpene hydrocarbons and phenolic monoterpenes, but the concentration of these compounds varied greatly among the oils examined. The results of the antimicrobial assay showed that essential oils extracted from Sardinian Thymus species have an antimicrobial activity comparable to the one observed in other thyme oils. It seems also confirmed that the antimicrobial properties of thyme essential oils are mainly related to their high phenolic content. Among the single compounds tested carvacrol and thymol turned out to be the most efficient against both reference strains and food-derived bacteria. The results of this study confirmed the possibility of using thyme essential oils or some of their components in food systems to prevent the growth of foodborne bacteria and extend the shelf-life of processed foods.

Anti-Bacterial Agents↗

Effect of thermocycling on the shear bond strength of a cyanoacrylate orthodontic adhesive.

The purpose of this study was to evaluate the effects of thermocycling on the shear bond strength of a cyanoacrylate adhesive system, specifically 24 hours after bonding when the adhesive has achieved most of its bond strength and after thermocycling. Forty freshly extracted human molars were collected and stored in a solution of 0.1% (weight/volume) thymol. The teeth were cleaned, polished, and randomly separated into 2 groups: group I, cyanoacrylate adhesive debonded after 24 hours immersion in deionized water at 37 degrees C; and group II, cyanoacrylate adhesive debonded after thermocycling at 5 degrees C and 55 degrees C. The results of the t test comparing the 2 groups (t = 6.84) indicated significant differences between them (P =.0001). The cyanoacrylate adhesive at 24 hours had significantly greater shear bond strength (macro x = 7.1 +/- 3.3 MPa) than after thermocycling 500 times between 5 degrees C and 55 degrees C (macro x = 1.5 +/- 1.4 MPa). The findings indicated that the cyanoacrylate adhesive tested has clinically adequate shear bond strength at 24 hours after initial bonding but loses about 80% of its strength after thermocycling. The clinician should consider all properties of the adhesive, including no need for a curing light, working time of 5 seconds before the adhesive starts to set, and the significant decrease in bond strength after thermocycling.

Cyanoacrylates↗

Inhibition of hormone-sensitive adenyl cyclase by phenothiazines.

Chlorpromazine (3 x 10(-4)M) prevents the stimulation of adenyl cyclase activity in thyroid membranes produced by thyrotropin and prostaglandin, ACTH stimulation of adenyl cyclase in adrenal tissue, and glucagon- and epinephrine-stimulation of adenyl cyclase activity in liver. Baseline activity is unaffected. Parathyroid hormone stimulation of kidney preparations was not inhibited under these conditions. At chlorpromazine concentrations >3 x 10(-4)M F(-)-stimulated cyclase activity of thyroid and adrenal tissue was increased. Other phenothiazines, trifluoperazine, and prochlorperazine, have similar effects on thyrotropin and F(-)-stimulated cyclase activity of thyroid. Na(+)- K(+)-dependent ATPase of thyroid is also inhibited by chlorpromazine. Since thymol causes a similar dissociation of hormone- and F(-)-stimulated adenyl cyclase, it is concluded that the surface properties of these agents best account for their effects on adenyl cyclase.

Adenine Nucleotides↗

Effect of an antibacterial dental varnish on the levels of prostanoids, leukotriene B4, and interleukin-1 beta in gingival crevicular fluid.

The aim of this study was to investigate the effects of a chlorhexidine/thymol-containing dental varnish on the levels of prostaglandin E2 (PGE2), prostaglandin I2 (PGI2), leukotriene B4 (LTB4), and interleukin-1 beta (IL-1 beta) in gingival crevicular fluid (GCF). The material consisted of 15 adolescents undergoing treatment with fixed orthodontic appliances. Four buccal sites adjacent to bands or brackets and exhibiting a mild chronic gingival inflammation were selected in the upper quadrants of each patient. According to a split-mouth technique, the first and second quadrants were randomly treated with either a varnish (Cervitec) containing 1% chlorhexidine diacetate and thymol (CHX/thymol) or a placebo varnish without active ingredients. The varnishes were applied immediately after the baseline registration, and follow-up examinations were carried out after 3, 8, and 30 days. GCF was sampled with the aid of a paper strip and the volume was determined using a Periotron 8000. The concentrations of PGE2, PGI2, LTB4, and IL-1 beta in GCF were assessed using radioimmunoassay and ELISA techniques. The results unveiled statistically significant reductions of PGE2, PGI2, and LTB4 levels in GCF following the active varnish treatment when compared to baseline values. A slight drop in IL-1 beta levels was registered after both active and placebo varnish applications, but the differences were not significant. The results suggest that treatment with an antibacterial varnish decreases the levels of inflammatory mediators PGE2, PGI2, and LTB4 in gingival crevicular fluid and further support the concept that topical application of a CHX/thymol-containing varnish is beneficial in patients with chronic gingival inflammation.

Adolescent↗

A new method for the preparation of beta-amylase from sweet potato.

A simple method for the preparation of sweet potato beta-amylase by thymol amylose adsorption is described. The method is far more efficient and gives higher recovery of the enzyme. The crystalline enzyme thus obtained is found to be homogeneous by gel chromatography, polyacrylamide gel electrophoresis.

Adsorption↗

Effects of rapid cooling on the mechanical and electrical activities of smooth muscles of guinea pig stomach and taenia coli.

The effects of rapid cooling on the mechanical and electrical activities of the guinea pig taenia coli and circular muscle of the stomach were investigated. Lowering the temperature from 32 degrees to 10 degrees C (cold shock) depolarized the membrane and increased the membrane resistance in both tissues. However, in the taenia coli, an initial reduction of membrane resistance was observed. In both tissues, contracture evoked by cold shock and rapid relaxation after rewarming, preceded the changes of membrane properties. Displacements of the membrane potential did not modify the amplitude of contracture under cold shock. Caffeine and thymol modified the membrane properties, but the effects of cold shock were still observed. The effects of cold shock were also observed on K-induced contracture. It was postulated that at least two different sites of sequestered bound Ca are located in these smooth muscles and are responsible for evoking the mechanical response. One component possesses a close relation to membrane and the other component is presumably sequestered within the muscle.

Animals↗

Identification of susceptibility to malignant hyperpyrexia in swine.

In vitro muscle contracture responses in swine susceptible to malignant hyperpyrexia (MH) were similar to those found in muscle from humans susceptible to this anaesthetic complication, confirming the suitability of the pig as an animal model for studying MH. The results suggest that there are different degrees of susceptibility to MH. Whichever drug was used, there was some overlap in the contracture responses between susceptible animals and controls, suggesting that the most accurate way of identifying susceptibility to MH is to use a variety of chemical agents, the best of which seem to be halothane, caffeine, suxamethonium and potassium chloride. Thymol, which is used as a preservative in commercial preparations of halothane, potentiates halothane contractures, but it is not known if this is significant clinically.

Animals↗

Cytotoxic and nongenotoxic effects of phenolic compounds in human pulp cell cultures.

Phenolic compounds are widely used in clinical dentistry as sedatives for the dental pulp, as disinfectants for caries, and as root canal medications. The pathobiological effects of various phenolic compounds on human dental pulp fibroblasts were investigated with Hoechst 33258 fluorescence assay and DNA precipitation assay. All phenolic compounds showed cytotoxicity in Hoechst 33258 fluorescence assay by inhibiting cellular DNA in a concentration-dependent manner. The 50% inhibition concentrations required to decrease the cellular DNA contents by guaiacol, phenol, eugenol, and thymol were 9.8, 4.5, 0.9, and 0.5 mM, respectively. However these phenolic compounds did not cause DNA single-strand breaks in cultured human pulp fibroblasts. These results indicate that phenolic compounds are cytotoxic agents but are without genotoxic effects on human pulp fibroblasts in vitro. However care should be taken to reduce the possibility of pulpal as well as periapical irritations from inadvertent extrusion of these substances in clinical usage.

Bisbenzimidazole↗

Antifungal activity of the essential oil of Thymus pulegioides on Candida, Aspergillus and dermatophyte species.

The composition of the essential oil of Thymus pulegioides and its antifungal activity on Candida, Aspergillus and dermatophyte fungal strains were studied. Essential oil from the aerial parts of the plant was obtained by hydrodistillation and analysed by GC and GC-MS. The oil showed high contents of carvacrol and thymol. The MIC and minimal lethal concentration were used to evaluate the antifungal activity against Candida (seven clinical isolates and four ATCC type strains), Aspergillus [five clinical isolates, and two Colección Española de Cultivos Tipo (CECT) and two ATCC type strains] and five clinical dermatophyte strains. Antifungal activity was evaluated for the essential oil and for its main components. To clarify its mechanism of action on yeasts and filamentous fungi, flow-cytometric studies of cytoplasmic membrane integrity were performed, and the effect on the amount of ergosterol was investigated. Results showed that T. pulegioides essential oil exhibited a significant activity against clinically relevant fungi, mainly due to lesion formation in the cytoplasmic membrane and a considerable reduction of the ergosterol content. The present study indicates that T. pulegioides essential oil has considerable antifungal activity, deserving further investigation for clinical applications.

Antifungal Agents↗

Studies on the relationship between serum colloidal reactions (ZTT and TTT) and IgG subclasses, especially IgG1 and IgG2.

The relationship between zinc sulfate turbidity test (ZTT and thymol turbidity test (TTT), and IgG subclasses, especially IgG1 and IgG2, was studied. Serum colloidal reactions of specimens from patients with IgG-myeloma usually show abnormally high values in ZTT and low values in TTT. But in some cases, values in ZTT and TTT are both low, and in a few cases both values are abnormally high. In order to see the reason why IgG-myelomas are classified into these three groups according to serum colloidal reactions. IgG subclasses of monoclonal proteins (MPs) isolated by zone electrophoresis were determined by immunodiffusion with anti IgG-subclasses. Results revealed that IgG1-myeloma showed high ZTT and low TTT and IgG2-myeloma showed low ZTT and TTT. Only two out of our 29 cases with IgG-myeloma showed high ZTT and TTT. MP from one of them belonged to IgG1-kappa and that from the other to IgG2-lambda. We checked the relationship between turbidity tests and electrophoretic mobility of MP and also that between these tests and L-chains' type of MP, but did not find any relationship between them. These findings suggest that IgG1 innately reacts with ZTT but not with TTT, while IgG2 does not react with any of these tests. MP from cases showing high values in both ZTT and TTT may have a special idiotype.

Antibodies, Monoclonal↗

Mutans streptococci in saliva and interdental spaces after topical applications of an antibacterial varnish in schoolchildren.

The effect of a chlorhexidine-containing varnish on the levels of mutans streptococci in saliva and in interdental spaces was investigated in 33 15-year-old schoolchildren. Each child was treated with an active (1% chlorhexidine; 1% thymol) and a placebo varnish preparation following the split-mouth technique. The varnishes were applied twice in small amounts into upper interdental areas with a 2-d interval. Mutans streptococci in saliva and interdental plaque was sampled and enumerated during 3 months. The results showed an immediate reduction of the number of interdental mutans streptococci on both test and placebo side after the varnish applications. The levels, however, were significantly lower in the test quadrants compared with the placebo-treated sides after 8, 30 and 90 d. Thus, the findings indicate a slower recolonization in interdental spaces treated with the active preparation. The levels of mutans streptococci in saliva were significantly reduced 1 and 3 months after varnish treatment, suggesting a long-term effect of the antibacterial varnish.

Adolescent↗

The antimicrobial activity of essential oils and essential oil components towards oral bacteria.

A method for reproducibly determining minimal inhibitory concentrations and minimal bactericidal concentrations of plant extracts towards fastidiously and facultatively anaerobic oral bacteria, predicated upon measurements of optical densities in microtitre plate wells, was devised. The antimicrobial properties of some botanical oils were surveyed; of these, Australian tea tree oil, peppermint oil, and sage oil proved to be the most potent essential oils, whereas thymol and eugenol were potent essential oil components.

Actinomyces viscosus↗