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Inhibitory effects of spices on growth and toxin production of toxigenic fungi.

The inhibitory effects of 29 commercial powdered spices on the growth and toxin production of three species of toxigenic Aspergillus were observed by introducing these materials into culture media for mycotoxin production. Of the 29 samples tested, cloves, star anise seeds, and allspice completely inhibited the fungal growth, whereas most of the others inhibited only the toxin production. Eugenol extracted from cloves and thymol from thyme caused complete inhibition of the growth of both Aspergillus flavus and Aspergillus versicolor at 0.4 mg/ml or less. At a concentration of 2 mg/ml, anethol extracted from star anise seeds inhibited the growth of all the strains.

Aflatoxins↗

Inactivation of nocardiophages phi C and phi EC by extracts of bacteriophage-attachable cells.

Cultures of several species of Nocardia, including N. erythropolis Mat-Ce and Mat-cE mating strains, were extracted with solvents in an attempt to isolate an inactivating complex for nocardiophages phiC and phiEC. Ethanol was the only solvent found effective in solubilizing an inhibitory substance. Inactivating extracts were obtained from the cells of all species to which the phage were able to attach. After extraction of whole cells or cell wall preparations, the phage could not effectively attach to them. Both phages phiC and phiEC were inactivated by the same complex. However, phage phiEC inactivation was 10-fold greater than phiC inactivation. The velocity of inactivation was about 4.1 x 10(2) plaque-forming units per microgram per minute for phiC and 1.1 x 10(3) plaque-forming units per microgram per minute for phage phiEC. The cell extracts required divalent cations for phage inactivation. The inhibitory capacity of the cell extracts was reduced or lost by the activity of proteolytic enzymes, Tween 80, 2-mercaptoethanol, thymol, and sodium lauryl sulfate. Boiling the extract for 10 min did not alter its activity. The inactivating substance was postulated to be a lipoprotein of considerable complexity, unique in the ease with which it is solubilized from host cells by ethanol.

Adsorption↗

Study of the effect of Bronchipret on the lung function of five Austrian saddle horses suffering recurrent airway obstruction (heaves).

The effects of an oral preparation containing an extract of thyme and primula (Bronchipret; Bionorica) on the lung function of five horses suffering heaves were determined in a longitudinal study. The horses accepted the product well. The plasma concentrations of the marker substance, thymol, indicated that at least one of the substances in the extract had been absorbed from the gastrointestinal tract. The compliance, pulmonary pressure and airway resistance of the horses' lungs were all significantly improved after one month of treatment However, the severity of their clinical signs and their arterial oxygen partial pressure had not improved significantly.

Administration, Oral↗

Acute rat lung injury: feasibility of assessment with micro-CT.

PURPOSE: To evaluate the feasibility of micro-computed tomography (CT) for analysis of the lung fine structure and its alterations during endotoxin-induced lung injury. MATERIALS AND METHODS: Intravital perfusion-fixed rat lungs with (n = 5) and without (n = 5) endotoxin perfusion were scanned with micro-CT. Three imaging modalities (conventional histology, intravital microscopy, and electron microscopy) were used to document the effect of endotoxin and the in vivo application of contrast agent (a mixture of barium sulfate, gelatin, and thymol). The effect of endotoxin on structural changes of the lung was evaluated with analysis of variance. RESULTS: Intravital microscopy, conventional histology, and electron microscopy demonstrated capillary perfusion of contrast agent, inflated alveoli, and no extravasation of barium sulfate in the extravascular space. Systemic application of endotoxin led to a significant increase in the soft-tissue volume of the lungs (ie, tissue edema) (58.09 microm(3)+/- 4.6 [standard error of the mean] vs 8.31 microm(3)+/- 1.63, P <.001) and significant thickening of the alveolar walls (34.01 microm +/- 4.5 vs 14.83 microm +/- 2.5, P <.001) at micro-CT. Simultaneously, endotoxin-treated rat lungs showed a significant reduction in total air space (49.74 microm(3)+/- 1.72 vs 100.99 microm(3)+/- 1.16, P <.001). CONCLUSION: These findings indicate that micro-CT is feasible for structural evaluation of the lung fine structure and its alterations during endotoxin-induced lung injury.

Animals↗

Calcium-dependent resistance to stretch and stress relaxation in resting smooth muscles.

Mechanical responses to stretch and length-tension relations were examined in rabbit taenia coli, mesenteric vein, aorta, and myometrium and in guinea pig taenia coli made atonic by incubation in Krebs-bicarbonate solution at 20-22 degrees C. When stretched 10% of the length at which maximum active tension is observed (Lo) in 0.5 s, the muscles showed a transient large force (resistance to stretch) that decayed to a new constant level within minutes (stress relaxation). The resistance to stretch decreased markedly in Ca2+-free [disodium ethylene glycolbis-(beta-aminoethylether)-N,N-tetraacetic acid (EGTA)] Krebs but was restored in normal Krebs solution. Calcium removal did not affect the passive length-tension curve. The absence of Ca2+ did not change the steady-state force maintained by the muscle; thus stretch resistance was not due to tone. Blockade of Ca2+ influx associated with electrical activity with 5-[3,4-dimethoxyphenethyl)methylamino]-2-(3,4,5-trimethoxyphenyl-2-isopropylvaleronitrile (D-600) and of Ca2+ release from intracellular sites with thymol (1 mM) completely blocked contraction but did not alter the responses to stretch, thus dissociating the responses to stretch from these processes and tension development. The Ca2+-dependent stress relaxation showed a dependence on muscle length similar to that for active tension development. Except at long muscle lengths, where connective tissue markedly affects length-tension relations, most of the "viscoelasticity" of these smooth muscles is dependent on calcium and may be largely due to the straining of crossbridges that are attached, but not generating a net force, in the resting state.

Animals↗

Effect of chlorhexidine varnish and gel on mutans streptococci in margins of restorations in adults.

The aim of this study was to compare the effect of two different chlorhexidine regimens, varnish and gel, on mutans streptococci (MS) in the margins of restorations and in saliva. Eighteen subjects (mean age 44 years) with >250,000 MS/ml saliva and with many restored teeth were assigned to three groups, each comprising 6 individuals. They were treated with: (1) Cervitec varnish, containing 1% chlorhexidine and 1% thymol; (2) placebo varnish, or (3) Corsodyl gel, containing 1% chlorhexidine. The varnish was applied on two separate occasions to all tooth surfaces at an interval of 3-4 days. The gel was applied for 3 x 5 min in individual applicators on 2 consecutive days. Plaque samples from 8-9 margins of restorations per subject, mainly in the premolar and molar region, and stimulated saliva samples were collected at baseline and after 1, 4, 8 and 12 weeks. The general finding was that the Corsodyl gel gave the most pronounced reduction of MS followed by the Cervitec varnish. According to repeated measures ANOVA, no significant differences could be demonstrated either for MS in margins or in saliva among the three groups (p = 0.06 for margins and p = 0.07 for saliva). However, within the groups, the reduction of MS over time was statistically significant, both in margins and in saliva (p < 0.001 and p < 0.05, respectively). Student-Newman-Keuls test showed that MS in margins were significantly reduced at most sampling occasions in both chlorhexidine groups: the reduction was more pronounced in the Corsodyl gel group. The conclusion from this study is that the 1% chlorhexidine gel Corsodyl, administered 6 times during 2 days, is more effective in suppressing MS in the margins of restorations and in saliva than the 1% chlorhexidine varnish Cervitec, administered twice within 3-4 days.

Adult↗

Suppression of caries-related microorganisms in dentine lesions after short-term chlorhexidine or antibiotic treatment.

This study investigated the efficiency of a chlorhexidine varnish and an antibiotic paste in suppressing the cultivable microflora of deep dentine cavities in a stepwise excavation procedure. Subsequent to enamel preparation and removal of the central biomass, infected dentine was sampled from the cavity floor. Ten cavities each were either covered with the 1% chlorhexidine- and 1% thymol-containing varnish Cervitec (CE), the demeclocycline hydrocortisone-containing ointment Ledermix (LE) or received no treatment as control (CO). A compomer composite was used as intermediate restorative. Cavities were reassessed after 6 weeks and again dentine samples were microbiologically investigated for total viable counts, mutans streptococci and lactobacilli. After 6 weeks a significant reduction of the total viable counts was observed in the LE group (p = 0.011) compared to the control, whereas no differences were found in the CE group (p > 0.05). Mutans streptococci were rarely recovered at baseline and after 6 weeks. Compared to the CO group counts of lactobacilli were significantly reduced in the CE and LE groups (p < 0.05). Lactobacillus species were frequently recovered at baseline and after 6 weeks of observation. Lactobacillus rhamnosus was the predominant species in all samples investigated. Application of CE or LE resulted in reduced counts of lactobacilli after a period of 6 weeks. Although none of the materials completely eliminated the viable microorganisms, the use of LE was more effective than CE in reducing the total anaerobic microorganisms associated with carious dentine.

Actinomyces↗

Effect of an antimicrobial-containing varnish on root demineralisation in situ.

The effect of an antimicrobial-containing varnish on root demineralisation was investigated. The in situ demineralisation effect on the varnish, with or without the active ingredients chlorhexidine and thymol, was measured by means of microradiography in comparison with no application after a 2-week period in vivo. Furthermore, the effect of one or two applications was investigated by the 10 participants who carried sound intact roots in an appliance for four consecutive 2-week periods. In each period, different roots were mounted in the buccal flanges of a lower prosthesis, the experiment being of a randomized cross-over design. The results show that: (1) A single varnish treatment with active ingredients reduced lesion depth and mineral loss by about 77 and 82%, respectively. (2) The control varnish had no effect, and it can be concluded that antimicrobial-releasing varnishes are promising materials for root caries prevention.

Administration, Topical↗

Effect of chlorhexidine-containing varnish (Cervitec) on microbial vitality and accumulation of supragingival dental plaque in humans.

The effect on local plaque formation of a varnish containing 1% chlorhexidine and 1% thymol (Cervitec) was evaluated. Ten volunteers with clinically healthy oral conditions were asked to refrain from any kind of oral hygiene measures for three periods of three days. Undisturbed plaque formation was recorded during the first experimental period. At the beginning of the second period, the varnish was applied to six vestibular enamel surfaces and removed after 1 h. The third experimental period was initiated 12 weeks after varnish application to assess a potential long-term effect. During each period of plaque formation, samples were collected from the vestibular surfaces after 24 h (from teeth 15/25), after 48 h (from teeth 14/24) and after 72 h (from teeth 13/23), respectively, and evaluated for total microscopic bacterial counts (BC) and colony forming units (CFU). Microbial vitality was assessed by the plating efficiency [PE = (CFU/BC) x 100] and directly by a vital fluorescence (VF) technique. VF of 48- and 72-hour plaque was significantly reduced after Cervitec application. An inhibitory effect by Cervitec could not be discerned 12 weeks after varnish treatment.

Adult↗

Effect of an antibacterial varnish on mutans streptococci in plaque from enamel adjacent to orthodontic appliances.

The effect of an antibacterial varnish (Cervitec) on the levels of mutans streptococci in plaque adjacent to bonded orthodontic brackets was evaluated in 18 children using a split-mouth technique with a placebo varnish control. The test varnish contained 1% chlorhexidine and 1% thymol as active ingredients. Both varnishes were applied on four occasions during a 3-month period, and plaque was subsequently collected between 1 week and 6 months after the onset of treatment. All teeth involved in the study were carefully examined and clinically assessed for enamel demineralization prior to onset of the fixed appliances and immediately after debonding. The results showed a more frequent growth of mutans streptococci in the dental plaque collected from placebo-treated quadrants as compared with the test quadrants on all sampling occasions. The proportion of mutans streptococci within the plaque microflora was significantly (p < 0.05-0.01) lower on the test sides than on the opposite sides at the 1-week and 1-month examinations. The incidence of incipient enamel lesions around the brackets and along the gingival margin was generally low, and no differences were found between the test and placebo varnish treated quandrants at the time of debonding. The results suggest that mutans streptococci in plaque from orthodontic patients can be suppressed effectively by topical applications of an antibacterial varnish.

Adolescent↗

Protection of bovine enamel and dentine by chlorhexidine and fluoride varnishes in a bacterial demineralization model.

In an in vitro demineralization model, the protective effect of two chlorhexidine varnishes, Cervitec (1% w/w chlorhexidine diacetate and 1% w/w thymol) and EC40 (40% w/w chlorhexidine diacetate), was compared with that of Fluor Protector, a varnish containing 0.1% w/w F. The demineralization model comprised an acidogenic Streptococcus mutans suspension in agarose placed on enamel or dentine specimens. The experiments extended over three serial 22-hour demineralization periods with fresh S. mutans suspensions for each period. To determine whether the varnishes released enough demineralization-inhibiting compounds, approximately 10 microliters of the varnishes was applied adjacent to the enamel and dentine specimens just before the first application of the S. mutans suspensions and left during the serial experiments (release study). In a separate series of experiments, the effect of the pretreatment of the enamal and dentine specimens with the various varnishes was tested (pretreatment study). In the release study the protective effect for enamel decreased in the order: EC40 = Fluor Protector >> Cervitec = no treatment. For dentine this order was: EC40 >> Fluor Protector = Cervitec > no treatment. In the pretreatment study, the enamel specimens were best protected by Fluor Protector (Fluor Protector >> Cervitec = EC40 > no treatment), while the dentine specimens were best protected by the chlorhexidine treatments (Cervitec = EC40 > Fluor Protector > no treatment). A 1:1 mixture of Cervitec and Fluor Protector was as effective as the most effective component alone. It is concluded that a varnish containing both fluoride and chlorhexidine may be useful, since it could give optimal protection to both enamel and dentine.

Animals↗

Effect of catalase on endotoxin-induced acute lung injury in unanesthetized sheep.

Administration of endotoxin intravenously to unanesthetized sheep causes an acute lung injury characterized by increased microvascular barrier permeability and subsequent pulmonary edema. Endotoxin-induced sheep lung injury can be attenuated by leukocyte depletion, and may be mediated by toxic metabolites of oxygen. We studied effects of administering catalase, which catalyzes conversion of hydrogen peroxide to oxygen and water, to sheep subsequently infused with endotoxin to test the hypothesis that hydrogen peroxide plays a role in the pathogenesis of lung injury. We found that infusions of endotoxin (1 microgram/kg) into untreated sheep caused the expected biphasic response, a transient, early, marked pulmonary arterial hypertension followed by a prolonged increase in protein-rich lung lymph flow characteristic of increased microvascular permeability filtration in the lungs. Intraperitoneal injections of catalase (50 mg/kg) prior to infusing endotoxin in these same sheep resulted in substantial catalase activity in plasma and in lung lymph, and attenuated the expected changes in pulmonary arterial pressure, lung lymph flow, and arterial leukocyte counts and oxygen tension after endotoxin infusions. Furthermore, mechanical elevation of hydrostatic pressure in the lungs of a catalase-treated sheep infused with endotoxin resulted in increased lung lymph flow with a decreased protein concentration, indicating that the microvascular barrier to fluid and protein was functionally intact. Administration of catalase that was inactivated by reaction with hydrogen peroxide in the presence of aminotriazole or administration of the catalase vehicle, thymol, had no effects on the sheep responses to endotoxin. We conclude that hydrogen peroxide plays a role in the pathogenesis of endotoxin-induced acute lung injury in sheep.

Acute Disease↗

Pharmacological studies on the anti-inflammatory action of phenolic compounds.

The mechanism of the anti-inflammatory action of phenolic compounds was examined using neutrophil chemotaxis. Chemotactic activity of guinea pig peritoneal neutrophils to N-formylmethionyl-leucylphenylalanine (FMLP) was suppressed in a concentration-dependent manner. The order of drug potency in inhibiting the neutrophil chemotaxis was eugenol much greater than thymol greater than guaiacol much greater than phenol. The concentrations of phenolic compounds used in these experiments did not induce lactate dehydrogenase (LDH) release and did not affect neutrophil viability. There was a consistent positive relation between the ID50 of superoxide anion generation in neutrophils and the inhibitory dose for neutrophil chemotaxis by phenolic compounds. A free phenolic hydroxyl group is essential for scavenging oxygen free-radicals and is also essential for inhibiting leukocyte chemotaxis, as was demonstrated in these experiments. These findings suggest that inhibition of leukocyte chemotaxis may be involved in the anti-inflammatory action of phenolic compounds, and that one of the anti-inflammatory actions of phenolic compounds is the prevention of the production of oxygen free-radicals by leukocytes.

Animals↗

Fatal large-volume mouthwash ingestion in an adult: a review and the possible role of phenolic compound toxicity.

OBJECTIVE: To describe a case of fatal mouthwash ingestion and review possible sources of toxicity. DESIGN: Case report. SETTING: Veterans Administration Medical Center. PATIENT: Single patient with massive mouthwash ingestion. MAIN RESULTS: This patient was a 45-year-old man who developed cardiovascular collapse and multiorgan system failure following a massive ingestion of mouthwash (almost 3 liters). His presentation was remarkable for a profound anion-gap metabolic acidosis and a significant osmolar gap. No other co-ingestants were identified, and he expired despite full supportive care including dialysis and mechanical ventilation. An autopsy failed to identify any other cause of death. Nonalcoholic ingredients of this mouthwash are phenolic compounds (eucalyptol, menthol, and thymol), and large-volume mouthwash ingestion will produce exposure in the reported toxic range of these ingredients. CONCLUSIONS: When ingested in large quantities, the phenolic compounds in mouthwash may contribute to a severe anion-gap metabolic acidosis and osmolar gap, multiorgan system failure, and death. These compounds, in addition to alcohol, may account for the adverse effects associated with massive mouthwash ingestion.

Acidosis↗

Cytasters induced within unfertilized sea-urchin eggs.

Conditions that induce the formation of asters in unfertilized sea-urchin eggs have been investigated. Monasters were formed by treatment of eggs with acidic or basic sea-water, or procaine- or thymol-containing sea-water. A second treatment step, incubation with D2O-containing, ethanol-containing or hypertonic sea-water induced multiple cytasters. The number and size of cytasters varied according to the concentration of agents and duration of the first and second treatments, and also upon the species of eggs and the season in which the eggs were obtained. Generally, a longer second treatment or a higher concentration of the second medium resulted in a higher number of cytasters per egg. Asters were isolated and then examined by light and electron microscopy. Isolated monasters apparently lacked centrioles, whereas cytasters obtained from eggs undergoing the two-step treatment contained one or more centrioles. Up to eight centrioles were seen in a single aster; the centrioles appeared to have been produced during the second incubation. Centrospheres prepared from isolated asters retained the capacity to nucleate the formation of microtubules in vitro as assayed by light and electron microscopy. Many microtubules radiated from the centre of isolated asters, whether they contained centrioles or not. This observation is consistent with many other reports that microtubule-organizing centres need not contain centrioles.

Animals↗

New diacylated 2-hydroxythymol derivatives from Melampodium divaricatum.

Melampodium divaricatum is a medicinal plant, which occurs in Central America. In a recent paper we reported the occurrence of acylated 2-hydroxy thymol glycosides as main constituents in this plant. This paper deals with the isolation of two new 2,5-dihydroxythymol ester derivatives. The formerly reported sesquiterpene lactone mikanokryptin was not found in our plant material.

Acylation↗

In vitro antiplaque effects of antiseptic phenols.

Several phenols known to be antiseptics were tested in vitro for their ability to adsorb to saliva-coated enamel and subsequently inhibit plaque formation. 3,5,4'-tribromosalicylanilide was found to be effective against growth and plaque formation of A viscosus, A naeslundii, S mutans and S sanguis. Dibromsalicil was effective against A viscosus. The other phenols (hexylresorcinol, thymol, phenylphenol and zinc phenolsulfonate) did not inhibit in vitro growth or plaque formation.

Animals↗

A simple and rapid in situ preconcentration method for trace ammonia nitrogen in environmental water samples using a solid-phase extraction followed by spectrophotometric determination.

A simple and rapid in situ preconcentration method for the spectrophotometric determination of trace ammonia nitrogen in environmental water samples has been developed based on solid-phase extraction using a small column packed with octadecyl group-bonded silica gel (Sep-Pak C18 cartridge). A water sample was taken into a graduated syringe for easy and simple operation and prevention of contamination immediately after sample collection. Ammonia in the sample was reacted with hypochlorite and thymol to be converted into indothymol blue; then the formed indothymol blue was collected as an ion pair between indothymol blue and tetrabutylammonium ion on a Sep-Pak C18 cartridge. The indothymol blue on the cartridge was stable for 4 days. The retained indothymol blue was easily eluted with a mixture of methanol and 0.01 mol/l sodium hydroxide solution. The color intensity due to the indothymol blue was spectrophotometrically measured at 725 nm. The proposed method was successfully applied to environmental water samples such as river water.

Ammonia↗