Relevant essential oil components: a minireview on increasing applications and potential toxicity.

Phenolic compounds carvacrol, thymol, eugenol, and vanillin are four of the most thoroughly investigated essential oil components given their relevant biological properties. These compounds are generally considered safe for consumption and have been used in a wide range of food and non-food applications. Significant biological properties, including antimicrobial, antioxidant, analgesic, anti-inflammatory, anti-mutagenic, or anti-carcinogenic activity, have been described for these components. They are versatile molecules with wide-ranging potential applications whose use may substantially increase in forthcoming years. However, some in vitro and in vivo studies, and several case reports, have indicated that carvacrol, thymol, and eugenol may have potential toxicological effects. Oxidative stress has been described as the main mechanism underlying their cytotoxic behavior, and mutagenic and genotoxic effects have been occasionally observed. In vivo studies show adverse effects after acute and prolonged carvacrol and thymol exposure in mice, rats, and rabbits, and eugenol has caused pulmonary and renal damage in exposed frogs. In humans, exposure to these three compounds may cause different adverse reactions, including skin irritation, inflammation, ulcer formation, dermatitis, or slow healing. Toxicological vanillin effects have been less reported, although reduced cell viability after exposure to high concentrations has been described. In this context, the possible risks deriving from increased exposure to these components for human health and the environment should be thoroughly revised.

Prospective randomized controlled blinded clinical trial evaluating biomarkers of acute kidney injury following 6% hydroxyethyl starch 130/0.4 or Hartmann's solution in dogs.

OBJECTIVE: To evaluate the effect of 6% hydroxyethyl starch (HES) 130/0.4, compared with a Hartmann's solution control (CRYST), on urine biomarkers of acute kidney injury (AKI) in dogs prescribed a fluid bolus. DESIGN: Randomized, controlled, blinded clinical trial January 2018 to February 2019. SETTING: University teaching hospital. ANIMALS: Forty client-owned dogs. INTERVENTIONS: Dogs prescribed a fluid bolus were randomized to receive at least 10 mL/kg of HES or CRYST with clinicians and investigators blinded to fluid type. Study fluid was used for further boluses as required in the following 24 hours, to a limit of 40 mL/kg total, after which fluid administration was open-label. MEASUREMENTS AND MAIN RESULTS: Urine was collected prior to and 6, 12, and 24 hours after the first study fluid bolus. Urine concentrations of AKI biomarkers: neutrophil gelatinase-associated lipocalin (NGAL), cystatin C, kidney injury molecule-1 (KIM), clusterin, and osteopontin were measured using a magnetic bead multiplexed assay. Osmolality-indexed biomarker concentrations were compared between groups over time with linear mixed-effects models, with P < 0.05 considered significant. The mean volume of study fluid administered was not significantly different between groups (HES: 23.1 mL/kg, CRYST: 25.9 mL/kg; P = 0.47, t-test). There were no significant differences between groups in change over time of osmolality-indexed urine concentrations of NGAL (P = 0.91), cystatin C (P = 0.95), KIM (P = 0.77), clusterin (P = 0.63), or osteopontin (P = 0.91). The maximum Veterinary Acute Kidney Injury (VAKI) score up to 7 days during hospitalization (P = 1.0) and in-hospital mortality (P = 0.49) were not significantly different between groups, as compared by Fisher's exact test. CONCLUSIONS: There were no differences in change over time of urine AKI biomarkers in dogs treated with 10 - 40 mL/kg HES or CRYST over 24 hours. Larger clinical trials with patient-centered outcomes are required to investigate the safety of HES in dogs.

Carvacrol and Thymol Attenuate Cytotoxicity Induced by Amyloid ß25-35 via Activating Protein Kinase C and Inhibiting Oxidative Stress in PC12 Cells.

Background: Our previous findings indicated that carvacrol and thymol alleviate cognitive impairments caused by Aß in rodent models of Alzheimer's disease (AD). In this study, the neuroprotective effects of carvacrol and thymol against Aß25-35-induced cytotoxicity were evaluated, and the potential mechanisms were determined. Methods: PC12 cells were pretreated with Aß25-35 for 2 h, followed by incubation with carvacrol or thymol for additional 48 h. Cell viability was measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. A flurospectrophotometer was employed to observe the intracellular reactive oxygen species (ROS) production. Protein kinase C (PKC) activity was analyzed using ELISA. Results: Our results indicated that carvacrol and thymol could significantly protect PC12 cells against Aß25-35-induced cytotoxicity. Furthermore, Aß25-35 could induce intracellular ROS production, while carvacrol and thymol could reverse this effect. Moreover, our findings showed that carvacrol and thymol elevate PKC activity similar to Bryostatin-1, as a PKC activator. Conclusion: This study provided the evidence regarding the protective effects of carvacrol and thymol against Aß25­35-induced cytotoxicity in PC12 cells. The results suggested that the neuroprotective effects of these compounds against Aß25-35 might be through attenuating oxidative damage and increasing the activity of PKC as a memory-related protein. Thus, carvacrol and thymol were found to have therapeutic potential in preventing or modulating AD.

Impact of sublethal exposure to synthetic and natural acaricides on honey bee (Apis mellifera) memory and expression of genes related to memory.

Acaricides are used by beekeepers in honey bee (Apis mellifera L.) colonies to control parasitic mites, but may also have adverse effects to honey bees. In this study, five commonly used acaricides were tested for their sublethal effects on memory and expression of neural-related genes in honey bees. Memory measured with the proboscis extension reflex (PER) assay was significantly reduced by topical treatment of bees with a single LD05 dose of formic acid at 2 and 24 h post treatment (hpt). However, tau-fluvalinate, amitraz, coumaphos, and formic acid, but not thymol, resulted in memory loss at 48 hpt. The LD05 doses of the acraricides did not affect expression of neuroligin-1, related to memory, or expression of major royal jelly protein-1, related to both memory and development, although expression of both genes was affected at LD50 doses. The LD05 doses of thymol, formic acid, amitraz and coumaphos increased defensin-1 expression, which is related to both memory and immunity. The effect of thymol, however, may have been due to its impact on the immune response rather than memory. This study demonstrates that acaricides vary in their effects on bee's memory, and that the widely used acaricide, formic acid, is particularly damaging.

Ayapana triplinervis Essential Oil and Its Main Component Thymohydroquinone Dimethyl Ether Inhibit Zika Virus at Doses Devoid of Toxicity in Zebrafish.

Zika virus (ZIKV) is an emerging mosquito-borne virus of medical concern. ZIKV infection may represent a serious disease, causing neonatal microcephaly and neurological disorders. Nowadays, there is no approved antiviral against ZIKV. Several indigenous or endemic medicinal plants from Mascarene archipelago in Indian Ocean have been found able to inhibit ZIKV infection. The purpose of our study was to determine whether essential oil (EO) from Reunion Island medicinal plant Ayapana triplinervis, whose thymohydroquinone dimethyl ether (THQ) is the main component has the potential to prevent ZIKV infection in human cells. Virological assays were performed on human epithelial A549 cells infected with either GFP reporter ZIKV or epidemic viral strain. Zebrafish assay was employed to evaluate the acute toxicity of THQ in vivo. We showed that both EO and THQ inhibit ZIKV infection in human cells with IC50 values of 38 and 45 µg/mL, respectively. At the noncytotoxic concentrations, EO and THQ reduced virus progeny production by 3-log. Time-of-drug-addition assays revealed that THQ could act as viral entry inhibitor. At the antiviral effective concentration, THQ injection in zebrafish does not lead to any signs of stress and does not impact fish survival, demonstrating the absence of acute toxicity for THQ. From our data, we propose that THQ is a new potent antiviral phytocompound against ZIKV, supporting the potential use of medicinal plants from Reunion Island as a source of natural and safe antiviral substances against medically important mosquito-borne viruses.

Thymol Affects Congruency Between Olfactory and Gustatory Stimuli in Bees.

Honey bees learn to associate sugars with odorants in controlled laboratory conditions and during foraging. The memory of these associations can be impaired after exposure to contaminants such as pesticides. The sub-lethal effects of acaricides such as 5-methyl-2-(propan-2-yl)-phenol (thymol) introduced into colonies to control varroa mites are of particular concern to beekeeping, due to detrimental effects of some acaricides on bees. Here we assess whether various odorant/sugar pairs are identically memorized in a differential appetitive olfactory conditioning experiment and whether this learning is affected by thymol exposure. Responses to odorants in retrieval tests varied according to the sugar they were paired with, a property called congruency. Interestingly, congruency was altered by pre-exposure to some thymol concentrations during retrieval tests, although electroantennography recordings showed it left odorant detection intact. This highlights the importance of taking into account subtle effects such as odor/sugar congruency in the study of the effect of pesticides on non-target insects, in addition to the simpler question of memory impairment.

Long-term dynamics of honey bee colonies following exposure to chemical stress.

Pesticide residues have been linked to reduced bee health and increased honey bee colony failure. Most research to date has investigated the role of pesticides on individual honey bees, and it is still unclear how trace levels of pesticides change colony viability and productivity over seasonal time scales. To address this question we exposed standard bee colonies to chemical stressors known to have negative effects on individual bees, and measured the productivity of bee colonies across a whole year in two environments: near Tucson Arizona and Sydney Australia. We exposed hives to a trace amount of the neonicotinoid imidacloprid and to the acaricide thymol, and measured capped brood, bee and honey production, as well as the temperature and foraging force of the colonies. The effect of imidacloprid on colony dynamics differed between the two environments. In Tucson we recorded a positive effect of imidacloprid treatment on bee and brood numbers. Thymol was associated with short-term negative effects on bee numbers at both locations, and may have affected colony survival at one location. The overall benefits of thymol for the colonies were unclear. We conclude that long-term and colony-level measures of the effects of agrochemicals are needed to properly understand risks to bees.

Comparison of the Potential Discoloration Effect of Bioaggregate, Biodentine, and White Mineral Trioxide Aggregate on Bovine Teeth: In Vitro Research.

INTRODUCTION: Tricalcium silicate cements can be used for pulp capping, pulpotomies, apical barrier formation in teeth with open apices, repair of root perforations, regenerative endodontics, and root canal filling. The aim of this study was to evaluate and compare the discoloration potential of 3 different tricalcium cements using a bovine tooth model. METHODS: Forty bovine anterior teeth have been used for the study. Crowns separated from the roots were randomly divided into 4 groups: the BioAggregate (IBC, Vancouver, Canada) group, the Biodentine (Septodont, Saint Maur des Fosses, France) group, the mineral trioxide aggregate Angelus (Angelus, Londrina, PR, Brazil) group, and the only blood group. Materials have been placed to the standardized cavities on the lingual surfaces of the crowns, and their contact with blood has been provided. The color values of the samples were measured with a digital tooth shade determinator (VITA Easyshade; VITA Zahnfabrik, Bad Sackingen, Germany) before the placement of the materials, after the placement of the materials, in the 24th hour, in the first week, in the first month, in the third month, and in the first year. The mean value of all groups was compared using the Tukey multiple comparison test (α = 0.05). RESULTS: All groups displayed increasing discoloration during a period of the first year. The "only blood group" showed the highest color change values, and it was followed as BioAggregate, mineral trioxide aggregate Angelus, and Biodentine, respectively. Statistically significant differences were found for Biodentine when compared with the only blood and BioAggregate groups (P < .05). CONCLUSIONS: Considering the results of the study, Biodentine is found to have the least discoloration potential among the tested materials.

Effect of a thymol application on olfactory memory and gene expression levels in the brain of the honeybee Apis mellifera.

Essential oils are used by beekeepers to control the Varroa mites that infest honeybee colonies. So, bees can be exposed to thymol formulations in the hive. The effects of the monoterpenoid thymol were explored on olfactory memory and gene expression in the brain of the honeybee. In bees previously exposed to thymol (10 or 100 ng/bee), the specificity of the response to the conditioned stimulus (CS) was lost 24 h after learning. Besides, the octopamine receptor OA1 gene Amoa1 showed a significant decrease of expression 3 h after exposure with 10 or 100 ng/bee of thymol. With the same doses, expression of Rdl gene, coding for a GABA receptor subunit, was not significantly modified but the trpl gene was upregulated 1 and 24 h after exposure to thymol. These data indicated that the genes coding for the cellular targets of thymol could be rapidly regulated after exposure to this molecule. Memory and sensory processes should be investigated in bees after chronic exposure in the hive to thymol-based preparations.

[Specific features of materials for initial pulpitis treatment].

The aim of the current study was to assess antibacterial and cytotoxic properties of Biodentine (Septodont), Rootdent (TehnoDent) and adhesive Futurabond НР (Voco). Two lines of experiments were carried out using cements water solutions and firm tablet-like samples (made by means of special pattern). Citotoxic activity was tested on NCTC L929 mice line fibroblasts culture. All the examined materials showed antibacterial activity against E. coli, S. aureus, C. albiсans, St. faecalis, mostly evident in Futurabond and the poorest in Biodentine samples. As for cytotoxic properties, Biodentine proved not to suppress metabolic activity stimulating odontotropic impact. The results confirm the analyzed materials to be a useful tool for deep caries lesions and initial pulpitis treatment.

Superficial skin necrosis and neurological complications following administration of local anaesthetic: a case report.

Local anaesthesia is used routinely throughout dentistry. Complications are, however, relatively uncommon. A unique case of superficial skin necrosis and neurological symptoms following administration of local anaesthesia for dental treatment is reported and the possible mechanisms behind this unusual presentation are discussed. Awareness of this possible complication and its mechanism helps clinicians diagnose and manage patients with similar problems appropriately.

Effect of endodontic sealers on tooth color.

OBJECTIVES: One of the goals of endodontic treatment is the adequate filling of the root canal,which is often done using gutta-percha and sealer. It has been reported that sealer remnants in the coronary pulp chamber cause tooth color changes. Therefore, this study was designed to examine the effect of endodontic sealer remnants on tooth color, testing the hypothesis that sealers cause coronal color changes. METHODS: Forty single-rooted human teeth were endodontically treated leaving excess sealer material in the coronary pulp chamber. The specimens were divided into four groups (n = 10) according to the endodontic sealer used (AH, AH Plus; EF, Endofill; EN,endome´ thasoneN; and S26, Sealer 26). Teeth were stored at 37 8C moist environment.Color coordinates (L*a*b*) were measured with a spectrophotometer before endodontic treatment(baseline-control), 24 h and 6 months after treatment. L*a*b* values were used to calculate color changes (DE). Data were statistically analyzed using Kruskal­Wallis and Mann­Whitney-U tests. RESULTS: Color changes were observed for all groups with S26 and EN producing the greatest mean DE values after 6 months. CONCLUSION: Endodontic sealer remnants affect tooth color confirming the experimental hypothesis. CLINICAL SIGNIFICANCE: This study examined the effect of endodontic sealer remnants on tooth color, and observed that after 6 months, the sealers produced unacceptable color changes.

Synthesis and evaluation of mutual prodrugs of ibuprofen with menthol, thymol and eugenol.

The present works deals with simple and efficient method of improving therapeutic efficacy of racemic ibuprofen by retarding gastrointestinal side effects through masking of carboxylic group chemically. This is achieved by synthesis and evaluation of ester derivatives of ibuprofen as mutual prodrugs with naturally occurring phenolic and alcoholic compounds. Promoieties like menthol; thymol and eugenol were selected with the aim of getting synergistic effect as these are natural analgesic having traditional medicinal values. Prodrugs are found to be highly lipophilic as compared to parent drug. All the prodrugs are found to be highly stable at acidic pH while undergoes hydrolysis at neutral and alkaline pH as indicated by their t(1/2) values. Synthesized prodrugs derivatives show increased anti-inflammatory activity that might be attributed to synergistic effect as ibuprofen conjugates to natural analgesics. Ulcer index shows much reduction in gastric ulceration compared to ibuprofen concluding the successful masking of acidic group.

In vitro antifungal activity of terpinen-4-ol, eugenol, carvone, 1,8-cineole (eucalyptol) and thymol against mycotoxigenic plant pathogens.

The aim of this study was to examine the effect of five naturally occurring compounds from essential oils on 10 different species of mycotoxigenic fungi involved in several plant diseases. The antifungal activities of terpinen-4-ol, eugenol, carvone, 1,8-cineole (eucalyptol) and thymol were observed in vitro on Fusarium subglutinans, Fusarium cerealis, Fusarium verticillioides, Fusarium proliferatum, Fusarium oxysporum, Fusarium sporotrichioides, Aspergillus tubingensis, Aspergillus carbonarius, Alternaria alternata and Penicillium sp. The naturally occurring compounds tested showed toxic effects on in vitro mycelium growth of all fungal species but with different level of potency. The results are encouraging for further investigations of in planta antifungal activities of these essential oils components.

Tissue reaction to Endométhasone sealer in root canal fillings short of or beyond the apical foramen.

OBJECTIVE: This study evaluated the response of periapical tissues to the endodontic sealer Endométhasone in root canal fillings short of or beyond the apical foramen. MATERIAL AND METHODS: Twenty root canals of premolars and incisors of 2 mongrel dogs were used. After coronal access and pulp extirpation, the canals were instrumented up to a size 55 K-file and the apical cemental barrier was penetrated with a size 15 K-file to obtain a main apical foramen, which was widened to a size 25 K-file. The canals were irrigated with saline at each change of file. The root canals were obturated either short of or beyond the apical foramen by the lateral condensation of gutta-percha and Endométhasone, originating 2 experimental groups: G1: Endométhasone/short of the apical foramen; G2: Endométhasone/beyond the apical foramen. The animals were killed by anesthetic overdose 90 days after endodontic treatment. The individual roots were obtained and serial histological sections were prepared for histomorphological analysis (H&E and Brown & Brenn techniques) under light microscopy. The following parameters were examined: closure of the apical foramen of the main root canal and apical opening of accessory canals, apical cementum resorption, intensity of the inflammatory infiltrate, presence of giant cells and thickness and organization of the apical periodontal ligament. Each parameter was scored 1 to 4, 1 being the best result and 4 the worst. Data were analyzed statistically by the Wilcoxon nonparametric tests (p=0.05). RESULTS: Comparing the 2 groups, the best result (p<0.05) was obtained with root canal filling with Endométhasone short of the apical foramen but a chronic inflammatory infiltrate was present in all specimens. CONCLUSIONS: Limiting the filling material to the root canal space apically is important to determine the best treatment outcome when Endométhasone is used as sealer.

Tissue reaction to Endométhasone sealer in root canal fillings short of or beyond the apical foramen

OBJECTIVE: This study evaluated the response of periapical tissues to the endodontic sealer Endométhasone in root canal fillings short of or beyond the apical foramen. MATERIAL AND METHODS: Twenty root canals of premolars and incisors of 2 mongrel dogs were used. After coronal access and pulp extirpation, the canals were instrumented up to a size 55 K-file and the apical cemental barrier was penetrated with a size 15 K-file to obtain a main apical foramen, which was widened to a size 25 K-file. The canals were irrigated with saline at each change of file. The root canals were obturated either short of or beyond the apical foramen by the lateral condensation of gutta-percha and Endométhasone, originating 2 experimental groups: G1: Endométhasone/short of the apical foramen; G2: Endométhasone/beyond the apical foramen. The animals were killed by anesthetic overdose 90 days after endodontic treatment. The individual roots were obtained and serial histological sections were prepared for histomorphological analysis (H&E and Brown & Brenn techniques) under light microscopy. The following parameters were examined: closure of the apical foramen of the main root canal and apical opening of accessory canals, apical cementum resorption, intensity of the inflammatory infiltrate, presence of giant cells and thickness and organization of the apical periodontal ligament. Each parameter was scored 1 to 4, 1 being the best result and 4 the worst. Data were analyzed statistically by the Wilcoxon nonparametric tests (p=0.05). RESULTS: Comparing the 2 groups, the best result (p<0.05) was obtained with root canal filling with Endométhasone short of the apical foramen but a chronic inflammatory infiltrate was present in all specimens. CONCLUSIONS: Limiting the filling material to the root canal space apically is important to determine the best treatment outcome when Endométhasone is used as sealer.

Age-related changes in the behavioural response of honeybees to Apiguard®, a thymol-based treatment used to control the mite Varroa destructor.

The parasitic mite Varroa destructor is responsible for heavy losses in honey bee colonies and represents a major threat to the beekeeping industry. Essential oils offer an attractive alternative to the use of synthetic chemicals for the control of varroa. Amongst them, thymol appears to be particularly promising. However, treatments using thymol as their active substance, such as the gel formulation Apiguard(®), are suspected to have adverse effects on honey bee colonies. In this study, laboratory assays are used to investigate the effects of Apiguard(®) exposure on honey bee behaviour. Our results reveal that honey bee responses to this anti-varroa treatment change with honey bee age. While 2-day-old bees respond neutrally to Apiguard(®), older bees generally avoid the Apiguard(®) gel. Responses of forager bees were particularly striking. Foragers appear to be repelled by Apiguard(®). Touching their antennae with Apiguard(®) induces robust fanning behaviour. Our data suggest, however, that forager bees exposed to Apiguard(®) in the hive can become habituated to this treatment. These results offer interesting new perspectives on the effects of Apiguard(®) on honey bee behaviour and serve to highlight age-related changes in honey bee responses to gustatory, as well as olfactory cues.

Contact dermatitis to Vicks VapoRub.

Vicks VapoRub (VVR) is a commonly used inhalant ointment that helps relieve symptoms of upper respiratory tract infections. It contains several plant substances, including turpentine oil, eucalyptus oil, and cedar leaf oil, which can potentially irritate or sensitize the skin, as well as camphor, menthol, nutmeg oil, and thymol. Although many reports describe allergic contact dermatitis (ACD) to the various constituents in VVR ointment, there are no cases of VVR directly causing ACD. We present a case of a patient who developed an ACD secondary to application of her VVR.

[Oxytocin and syndrome of inappropriate secretion of antidiuretic neonatal hormone. Case report of early severe hyponatremia and literature review]. / Oxitocina y sindrome de secreción inapropiada de hormona antidiurética neonatal. Reporte de un caso de hiponatremia severa temprana y revisión de la bibliografia.

This is a clinical case presentation of a full term newborn infant who suffered severe hyponatremia and early seizures, associated with maternal fluid overload with electrolyte free solutions and high doses of oxytocin for labor augmentation. Although this condition has been recognized since the 1960's with isolated reports, this particular case has features that needs further investigation, not only for the unsually severe hyponatremia, but most importantly we think, for the prominent signs of fluid retention, the infant had, that suggest excessive antidiuretic activity probably due to oxytocin. These findings are consistent with syndrome of inappropriate secretion of antidiuretic hormone. Although until now there is no proof that oxytocin by itself produces this syndrome. We think the association is possible in certain clinical circumstances, such as those found in this case. We also, briefly discussed the pathophysiology of perinatal hyponatremia, the neonatal treatment of this condition and the current guidelines for the women in labor. Hyponatremia should not be considered a benign condition, since in the neonate, it may affect brain function.

Surgical approach to iatrogenic complications of endodontic therapy: a report of 2 cases.

Iatrogenic complications in endodontics are not irrelevant. One of the most seen is overfilling of the root canal, which has a negative effect on prognosis for endodontically treated teeth. Although more than a half of the overfilled teeth heal satisfactorily after proper endodontic therapy, in some cases, such as injury of the inferior alveolar nerve or presence of filling material in soft tissues or sinus spaces, surgical procedure is more suitable and effective. The purpose of this article is to present 2 clinical cases of iatrogenic complications of endodontically treated teeth and the approach to them by surgical procedure. In the first case, extruded endodontic filling material was present in the periapical region of lower second premolar tooth causing anesthesia of the mental nerve; in the second case, extruded material was found to have perforated the cortical bone of maxilla and passed through the mucoperiosteum to the muscle fibers.