Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice / Eugenol e seu nanocarreador à base de lipossoma reduzem a ansiedade ao inibir a expressão de glioxalase 1 em camundongos

Abstract The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

Resumo A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.

Eugenol and its liposome-based nano carrier reduce anxiety by inhibiting glyoxylase-1 expression in mice / Eugenol e seu nanocarreador à base de lipossoma reduzem a ansiedade ao inibir a expressão de glioxalase 1 em camundongos

The most common form of psycho-social dysfunction is anxiety with depression being related closely without any age bar. They are present with combined state of sadness, confusion, stress, fear etc. Glyoxalase system contains enzyme named glyoxalase 1 (GLO1).It is a metabolic pathway which detoxifies alpha-oxo-aldehydes, particularly methylglyoxal (MG). Methylglyoxal is mainly made by the breakdown of the glycolytic intermediates, glyceraldehyde-3-phosphates and dihydroxyacetone phosphate. Glyoxylase-1 expression is also related with anxiety behavior. A casual role or GLO-1 in anxiety behavior by using viral vectors for over expression in the anterior cingulate cortex was found and it was found that local GLO-1 over expression increased anxiety behavior. The present study deals with the molecular mechanism of protective activity of eugenol against anxiolytic disorder. A pre-clinical animal study was performed on 42 BALB/c mice. Animals were given stress through conventional restrain model. The mRNA expression of GLO-1 was analyzed by real time RT-PCR. Moreover, the GLO-1 protein expression was also examined by immunohistochemistry in whole brain and mean density was calculated. The mRNA and protein expressions were found to be increased in animals given anxiety as compared to the normal control. Whereas, the expressions were decreased in the animals treated with eugenol and its liposome-based nanocarriers in a dose dependent manner. However, the results were better in animals treated with nanocarriers as compared to the compound alone. It is concluded that the eugenol and its liposome-based nanocarriers exert anxiolytic activity by down-regulating GLO-1 protein expression in mice.

A forma mais comum de disfunção psicossocial é a ansiedade intimamente relacionada com a depressão, sem qualquer barreira de idade. Elas estão presentes em um estado combinado de tristeza, confusão, estresse, medo etc. O sistema de glioxalase contém uma enzima chamada glioxalase 1 (GLO1). É uma via metabólica que desintoxica alfa-oxo-aldeídos, particularmente metilglioxal (MG). O metilglioxal é produzido principalmente pela quebra dos intermediários glicolíticos, gliceraldeído-3-fosfatos e fosfato de diidroxiacetona. A expressão da glioxalase 1 também está relacionada ao comportamento de ansiedade. Um papel casual ou GLO1 no comportamento de ansiedade usando vetores virais para superexpressão no córtex cingulado anterior foi encontrado e descobriu-se que a superexpressão local de GLO1 aumentava o comportamento de ansiedade. O presente estudo trata do mecanismo molecular da atividade protetora do eugenol contra o transtorno ansiolítico. Um estudo pré-clínico em animais foi realizado em 42 camundongos BALB / c. Os animais foram submetidos ao estresse por meio do modelo de contenção convencional. A expressão de mRNA de GLO1 foi analisada por RT-PCR em tempo real. Além disso, a expressão da proteína GLO1 também foi examinada por imuno-histoquímica em todo o cérebro e a densidade média foi calculada. Verificou-se que as expressões de mRNA e proteínas estavam aumentadas em animais que receberam ansiedade em comparação com o controle normal. Considerando que as expressões foram diminuídas nos animais tratados com eugenol e seus nanocarreadores baseados em lipossomas de forma dependente da dose. No entanto, os resultados foram melhores em animais tratados com nanocarreadores em comparação com o composto sozinho. Conclui-se que o eugenol e seus nanocarreadores baseados em lipossomas exercem atividade ansiolítica por regulação negativa da expressão da proteína GLO1 em camundongos.

Chemical composition and larvicidal activity of the essential oil of Pimenta dioica leaves / Composición química y actividad larvicida del aceite esencial de hojas de Pimenta dioica

In this study, we investigated the main constituent, the predominant class and biological activity of the essential oil extracted from the leaves of Pimenta dioica and the pattern of the major constituent against larvae in the third stage of Aedes aegypti. For this reason, we extracted the oil by hydrodistillation, identified its components by gas chromatography coupled with mass spectrometry (GC/MS) and calculated the lethal concentration (LC50) of the larvicidal activity using the Reed-Muench method. The results show that the oil consists mainly of eugenol, in which the phenylpropanoid class predominated and the lethal concentration, LC50, was 38.86 µg mL-1at a confidence level of 2.25 µg mL-1, while the eugenol standard presented LC5079.75 µg mL-1at a confidence level of 2.10 µg mL-1. Given the facts, we conclude that the oil is more active than the standard and that it has the potential to replace chemical larvicides.

En este estudio, investigamos el constituyente principal, la clase predominante y la actividad biológica del aceite esencial extraído de las hojas de Pimenta dioica y el patrón del constituyente principal contra las larvas en la tercera etapa de Aedes aegypti. Por este motivo, extrajimos el aceite por hidrodestilación, identificamos sus componentes mediante cromatografía de gases acoplada a espectrometría de masas (GC/MS) y calculamos la concentración letal (CL50) de la actividad larvicida mediante el método Reed-Muench. Los resultados muestran que el aceite está constituido principalmente por eugenol, en el que predominó la clase fenilpropanoide y la concentración letal, CL50, fue de 38,86 µg.mL-1 a un nivel de confianza de 2,25 µg.mL-1, mientras que el estándar de eugenol presentó CL50 79,75 µg.mL -1 a un nivel de confianza de 2,10 µg.mL-1. Dados los hechos, concluimos que el aceite es más activo que el estándar y que tiene el potencial de reemplazar los larvicidas químicos.

In vitro and in silico evaluation of the schistosomicidal activity of eugenol derivatives using biochemical, molecular, and morphological tools

Background Eugenol shows both antibacterial and antiparasitic activities, suggesting that it might be evaluated as an option for the treatment of praziquantel-resistant schistosome. Methods The in vitro activities of three eugenol derivatives (FB1, FB4 and FB9) on adult worms from Schistosoma mansoni were examined by fluorescence and scanning electron microscopy to analyze effects on the excretory system and integument damage, respectively. Biochemical tests with verapamil (a calcium channel antagonist) and ouabain (a Na+/K+-ATPase pump inhibitor) were used to characterize eugenol derivative interactions with calcium channels and the Na+/K+-ATPase, while in silico analysis identified potential Na+/K+-ATPase binding sites. Results The compounds showed effective doses (ED50) of 0.324 mM (FB1), 0.167 mM (FB4), and 0.340 mM (FB9). In addition, FB4 (0.322 mM), which showed the lowest ED50, ED90 and ED100 (p < 0.05), caused the most damage to the excretory system and integument, according to both fluorescence and scanning electron microscopy analysis. The death of adult worms was delayed by ouabain treatment plus FB1 (192 versus 72 hours) and FB9 (192 versus 168 hours), but the response to FB4 was the same in the presence or absence of ouabain. Besides, no changes were noted when all of the eugenol derivatives were combined with verapamil. Moreover, FB1 and FB9 inhibited Na+/K+-ATPase activity according to in silico analysis but FB4 did not show a time-dependent relationship and may act on targets other than the parasite Na+/K+-ATPase. Conclusion Eugenol derivatives, mainly FB4 when compared to FB1 and FB9, seem to act more effectively on the integument of adult S. mansoni worms.(AU)

Syzygium aromaticum como candidato a bactericida: efectos en la microbiota oral / Syzygium aromaticum as a bactericidal candidate: effects on the oral microbiota

El cavo de olor (Syzygium aromaticum) es un árbol, originario de Indonesia, con altura variable, pero que sobre pasa los 10 metros de altura, perteneciente a la familia de las Myrtaceae y cuyas flores que no han abierto, se convierten en botones, que al secar son los mencionados clavos de olor. Poseen como componente principal el Eugenol, entre otros compuestos orgánicos. Por sus características bioquímicas y organolépticas, le proporcionan varios beneficios para la salud, por actuar como estimulantes, antioxidante, con acción antibacterial, antiespasmódicas, además de su marcada acción analgésica y anestésica. Por su parte, la microbiota oral, está conformada por un amplio conjunto de microorganismos pertenecientes al ecosistema bucal y que a través del equilibrio de los mismos, se logrará un adecuado funcionamiento y desarrollo de las funciones fisiológicas en pro de la salud bucal del individuo. La presente investigación tiene como objetivo examinar los datos específicos en el uso del clavo de olor como agente bactericida en las afecciones bucodentales, encontrándose que si puede ser usado como agente bactericida por su marcado efecto sobre la microbiota oral a nivel de eliminar los microorganismos nocivos presentes en la misma, ya que actúa inhibiendo la recomposición de las proteínas, los ácidos nucleicos y la membrana de la pared celular, cambiando la permeabilidad de las células de los microorganismos, favoreciendo su muerte y a su ves favoreciendo el adecuado equilibrio de la microbiota oral, necesario para la adecuada salud bucodental(AU)

The clove (Syzygium aromaticum) is a tree, native to Indonesia, with variable height, but that exceeds 10 meters in height, belonging to the Myrtaceae family and whose flowers that have not opened, become buttons, that when drying are the aforementioned cloves. Their main component is Eugenol, among other organic compounds. Due to their biochemical and organoleptic characteristics, they provide several health benefits, for acting as stimulants, antioxidant, with antibacterial, antispasmodic action, in addition to their marked analgesic and anesthetic action. For its part, the oral microbiota is made up of a wide set of microorganisms belonging to the oral ecosystem and that through their balance, an adequate functioning and development of physiological functions will be achieved in favor of the oral health of the individual. The objective of this research is to examine the specific data on the use of cloves as a bactericidal agent in oral conditions, finding that it can be used as a bactericidal agent due to its marked effect on the oral microbiota at the level of eliminating harmful microorganisms present in it, since it acts by inhibiting the recomposition of proteins, nucleic acids and the cell wall membrane, changing the permeability of the cells of microorganisms, favoring their death and in turn favoring the proper balance of the oral microbiota, necessary for proper oral health(AU)

Eugenol and tricaine methanesulfonate as anhestesics for the pearl cichlid

Anesthesia reduces the handling process duration and prevent fish injuries. The anesthetic effect and ideal concentrations of eugenol and tricaine methanesulfonate (MS-222) were tested for pearl cichlid (Geophagus brasiliensis) juveniles with an average weight of 4.4 g in water at 24ºC. The criterion for determining the optimal dose considered an induction time of one minute. Experiment 1 tested the concentrations of 25, 75, 150 and 300 mg L-1 of eugenol. The best results were obtained at doses of 150 and 300 mg L-1. Experiment 2 aimed to establish a more accurate result by testing the concentrations of 180, 210, 240 and 270 mg L-1, and led to an estimation of 217 mg L-1 of eugenol to induce anesthesia in one minute. Experiment 3 evaluated 200, 300, 400, 500 and 600 mg L-1 of tricaine, of which the concentration of 294 mg L-1 was estimated to induce anesthesia in one minute. No significant differences were observed for recovery times when using either of the anesthetics. No mortality was observed within 24 hours after the experiments for any concentration of the anesthetics. The present study recommends 217 mg L-1 of eugenol or 394 mg L-1 of tricaine for anesthesia of the pearl cichlid.

Infiltration by Streptococcus mutans in provisional crowns fixed by two types of cementing agents ­ pilot double-blind randomized clinical trial / Infiltração por Streptococcus mutans em coroas dentárias provisórias fixadas por dois tipos de agentes cimentantes ­ estudo clínico randomizado duplo-cego piloto

Objective: When provisional acrylic crowns are used for a long time, they become more susceptible to marginal leakage by cariogenic bacteria. The objectives of this pilot clinical study were to compare cement based on zinc oxide-eugenol and calcium hydroxide by contamination with Streptococcus mutans, and calculate the sample size for the continuation of this study. Methods: Individuals receiving provisional crowns and following inclusion/exclusion criteria, were randomly distributed into 2 groups: zinc oxide-eugenol (n=8); calcium hydroxide (n=9). The temporary crowns were made by a blind researcher and cemented by another. Patients were also blinded by the cement used inside their crowns. After 2 months, a cement sample from the crowns' peripheral inner face was collected, placed in a tube containing 1 mL of sterile saline, serially diluted, plated on Mitis Salivarius Bacitracin agar, and incubated for 48 hours. Colony-forming units (CFU/mL) were counted. A statistical power analysis was performed to calculate sample size (1-ß=80%) and the Mann Whitney test to compare both cements (α=0.05). Results: Both cements were contaminated with S. mutans, with an average of 166.6 x 102 CFU/mL for calcium hydroxide and 435.3 x 102 CFU/mL for zinc oxide-eugenol, with no significant difference (p=0.311). The sample size calculated for this study was 36 per group. Conclusion: This pilot study suggests that there is important contamination inside provisional crowns used for two months, independent of the cement. The continuation of this study is needed, with a bigger sample size, to enable a comparison between the cements.

Objetivo: Quando coroas dentais provisórias são utilizadas por um longo período, elas se tornam susceptíveis à infiltração marginal por bactérias cariogênicas. O objetivo deste estudo clínico piloto foi comparar os cimentos a base de óxido de zinco e eugenol e hidróxido de cálcio pela contaminação com Streptococcus mutans e calcular o tamanho amostral para continuação deste estudo. Métodos: Indivíduos recebendo coroas provisórias e seguindo critérios de inclusão/exclusão, foram distribuídos aleatoriamente em dois grupos: óxido de zinco e eugenol (n=8); hidróxido de cálcio (n=9). As coroas provisórias foram feitas por um pesquisador cego e cimentadas por outro. Os pacientes também foram cegos quanto ao cimento utilizado dentro de suas coroas. Depois de 2 meses, amostras de cimentos foram coletadas da face interna periférica das coroas, colocadas em um tubo contendo 1 mL de solução salina estéril, diluídas de forma seriada, plaqueadas em ágar Mitis Salivarius Bacitracina e incubadas por 48 horas. Unidades formadoras de colônias (UFC/mL) foram contadas. Um teste de poder estatístico foi realizado para calcular o tamanho amostral (1-ß=80%) e o teste de Mann Whitney para comparar os dois cimentos (α=0.05). Resultados: Os dois cimentos foram contaminados com S. mutans, com uma média de 166.6 x 102 UFC/mL para o hidróxido de cálcio e 435.3 x 102 UFC/mL para o óxido de zinco e eugenol, com nenhuma diferença significativa (p=0.311). O cálculo amostral para este estudo foi 36 indivíduos por grupo. Conclusão: Este estudo piloto sugere que existe importante contaminação dentro de coroas provisórias utilizadas por 2 meses, independente do cimento. A continuação deste estudo é necessária, com maior tamanho amostral, para possibilitar a comparação entre os cimentos.

Effect of methyl eugenol on hypoxia/reoxygenation injury of human renal tubular epithelial cells and its mechanism / 中国中药杂志

This study aimed to investigate the effect of methyl eugenol(ME) on hypoxia/reoxygenation(H/R)-induced injury of human renal tubular epithelial HK-2 cells and its mechanism. The viability of HK-2 cells cultured with different concentrations of ME and exposed to H/R was detected by cell counting kit-8(CCK-8) assay. The effect of ME on the morphology of HK-2 cells was observed under an inverted microscope. The content of intracellular reactive oxygen species in different groups was detected after 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA) fluorescence staining. Cell apoptosis was determined by flow cytometry. Changes in mitochondrial membrane potential were monitored by JC-1 dye. The concentrations of nuclear factor erythroid 2 related factor 2(Nrf2), heme oxygenase-1(HO-1), and nicotinamide adenine dinucleotide phosphatase oxidase 4(Nox4) were measured by Western blot, followed by the assay of Nrf2 concentration changes in cytoplasm and nucleus by confocal fluorescence staining. The results showed that when the concentration of ME was 0-40 μmol·L~(-1), the activity of HK-2 cells was not affected. Compared with the model group, ME enhanced the activity of HK-2 cells and the cell morphology was normal. As revealed by further experiments, ME inhibited the release of reactive oxygen species and the decline in mitochondrial membrane potential of HK-2 cells after H/R injury, promoted Nrf2/HO-1 expression and Nrf2 translocation to the nucleus, and down-regulated the expression of Nox4, thereby significantly reducing apoptosis. This protective effect of ME could be reversed by the specific Nrf2 inhibitor ML385. These findings have preliminarily proved that ME effectively protected HK-2 cells against H/R injury, which might be related to its promotion of Nrf2/HO-1 signaling pathway and inhibition of Nox4. Such exploration on the possible mechanism of ME in the treatment of renal ischemia-reperfusion injury(IRI) and protection of organ function from the perspective of antioxidant stress has provided reference for related research on the treatment of acute kidney injury with traditional Chinese medicine.

Efeito Cardiodepressor do Acetato de Eugenil em Coração de Roedor / Cardiodepressive Effect of Eugenyl Acetate in Rodent Heart

Resumo No presente trabalho investigou-se o efeito inotrópico do acetato de eugenil (AE), bem como sua ação sobre a corrente de Ca2+ do tipo L (ICa,L). Os experimentos de contratilidade foram realizados em átrio esquerdo isolado de cobaia exposto às concentrações crescentes da droga (1 a 5.000μM). O AE reduziu a força de contração atrial (IC50=558±24,06μM) de modo dependente de concentração. O efeito do AE sobre a ICa,L também foi avaliado em cardiomiócitos ventriculares isolados de camundongos, utilizando-se a técnica de "patch-clamp". O AE apresentou um efeito inibitório (IC50=1.337±221μM) sobre os canais de Ca2+ sensíveis à voltagem (CaV1.2). Em conclusão, o AE apesenta efeito cardiodepressor que se deve, pelo menos em parte, à diminuição da entrada de Ca2+ nos cardiomiócitos.

Eugenol as an efficient anesthetic for neotropical fish Prochilodus nigricans (Teleostei, Prochilodontidae) / [Eugenol como um anestésico eficaz para o peixe neotropical Prochilodus nigricans (Teleostei, Prochilodontidae)]

The use of anesthetics in fish farming is essential to reduce stress during management. The present study proposes to evaluate the effect of eugenol as an anesthetic for the management of curimatã (Prochilodus nigricans). Fifty specimens were used (221.34±9.0 g; 25.8 ±1.1 cm), submitted to five treatments with concentrations of 0, 25, 50, 100 and 200 mgL−1 eugenol for 10 min. The times required to reach each anesthetic stage and the recovery of each fish were recorded. To check bloodglucose levels individual blood samples were taken from the fish before immersion and after ten minutes of anesthetic exposure. Evaluation of the anesthetic effect ongills was performed by histopathological analysis. The concentration of 50 mgL−1 was within the ideal limit of three minutes recommended for surgical anesthesia induction and did not significantly increase glucose levels when compared to the control group (35.7±19.4 mg dL−1), besides not causing gill injuries. At this concentration the fish presented controlled blood glucose levels within the basal levels, besides not taking the risk to have later problems concernig mortality, because this concentration ensures the good health of the gills and good recovery of the animals.(AU)

O uso de anestésicos em pisciculturas é indispensável para reduzir o estresse durante o manejo. O presente trabalho propôs avaliar o efeito do eugenol como anestésico para manejo do curimatã (Prochilodus nigricans). Foram utilizados 50 espécimes (221,34±9,0 g; 25,8 ±1,1 cm), submetidos a cinco tratamentos com concentrações de 0, 25, 50, 100 e 200 mg L−1 de eugenol em exposição de 10 minutos. Foram registrados os tempos necessários para atingir cada estágio anestésico e a recuperação de cada peixe. Para verificação dos níveis de glicose sanguínea, foram realizadas coletas individuais de sangue dos peixes antes da imersão e após os 10 minutos de exposição ao anestésico. A avaliação do efeito do anestésico sobre as brânquias foi realizada por meio de análises histopatológicas. A concentração de 50 mg L−1 ficou dentro do limite ideal de três minutos preconizado para indução à anestesia cirúrgica e não elevou significativamente os níveis de glicose quando comparada ao grupo controle (35,7±19,4 mg dL−1), além de não provocar lesões branquiais. Nessa concentração, os peixes apresentaram níveis de glicose sanguínea controlada, dentro dos teores basais, além de não correrem o risco de apresentarem problemas posteriores em relação à mortalidade, pois essa concentração garante a boa saúde das brânquias e uma boa recuperação dos animais.(AU)

Preparation of polymer nanoparticles loaded with Syzygium aromaticum essential oil: an oral potential application / Preparación de nanoparticulas polimericas cargadas con aceite esencial de Syzygium aromaticum: una potencial aplicación oral

Due to the biological activities of Syzygium aromaticum essential oil, its incorporation in methacrylate polymeric (Eudragit E100) nanoparticles (NP), physical characterization, and antimicrobial essays were evaluated. The clove bears great potential for applications in dentistry. The oil was obtained by hydrodistillation and oil loaded NP using the nanoprecipitation method. Particle size and polydispersity index were determined by photon correlation spectroscopy, and physical morphology by electron microscopy. Loading capacity and in vitro eugenol release were evaluated by gas mass chromatography, and the antimicrobial activity of oil loaded-NP was calculated against Streptococcus mutans. Different chemical ingredients were characterized, and eugenol was the principal compound with 51.55%. Polymer content was directly related to NP homogenous size, which was around 150 nm with spherical morphology. A 73.2% loading capacity of eugenol was obtained. Oil loaded NP presented a fickian-type release mechanism of eugenol. Antimicrobial activity to 300 µg/mL was obtained after 24 h.

Debido a las actividades biológicas del aceite esencial de Syzygium aromaticum, se evaluó su incorporación en nanopartículas (NP) de metacrilato polimérico (Eudragit E100), su caracterización y ensayos antimicrobianos. El clavo tiene un gran potencial para aplicaciones en odontología. El aceite se obtuvo por hidrodestilación y las NP cargado de aceite utilizando el método de nanoprecipitación. El tamaño de partícula y el índice de polidispersidad se determinaron mediante espectroscopia de correlación fotónica y su morfología por microscopía electrónica. La capacidad de carga y la liberación de eugenol in vitro se evaluaron mediante cromatografía de gases en masa, y la actividad antimicrobiana se evaluó contra Streptococcus mutans. Se caracterizaron diferentes ingredientes químicos, siendo el eugenol el principal compuesto con 51.55%. El contenido de polímero se relacionó directamente con el tamaño homogéneo de NP, que fue de alrededor de 150 nm con morfología esférica. Se obtuvo un 73,2% de capacidad de carga de eugenol. El aceite cargado en NP presentó un mecanismo de liberación de eugenol de tipo fickiano. La actividad antimicrobiana a 300 µg/mL se obtuvo después de 24 h.

Endodontic sealers and post-endodontic waiting time affect the bond strength of the fiber posts / Cimentos endodônticos e tempo de espera após endodontia afeta a resistência de união de pinos de fibra

Introduction: The resistance adhesive of a fiber post can be affected by several factors, such as the endodontic sealer and post-endodontic waiting time. Objective: The aim of this study was to evaluate the effect of different endodontic sealers and two different post-endodontic waiting times on the bond strength of fiber posts. Material and method: Seventy-two bovine teeth were endodontically treated and filled using three endodontic sealers: eugenol-based, epoxy resin-based, or mineral trioxide aggregate-based. The specimens were stored at 37°C for 24 hours or for 30 months. After the respective storage times, the root canals were prepared for luting fiber posts using RelyX U200. Push-out tests and analysis of failures were performed. The push-out data were analyzed by two-way analysis of variance to compare the effects of the endodontic sealer and with the t-test to compare the effects of post-endodontic waiting time. Result: The AH Plus sealer yielded the highest bond strength values at 30 months post-endodontics (11.26 Mpa) (p < 0.05), however no had difference with Endofill sealer at the same time. Endofill and MTA Fillapex sealers did not differ significantly in their effects, irrespective of the post-endodontic waiting time. Conclusion: In conclusion, the endodontic sealer used and post-endodontic waiting time affect the adhesive resistance of fiber posts. The adhesion increases significantly when the fiber post is cemented 30 months after the root canal filling, while the adhesion is reduced when cementing immediately after root canal treatment, in particular for eugenol-based endodontic sealers.

Introdução: A resistência de união de um pino de fibra pode ser afetada por vários fatores, como o cimento endodôntico e o tempo de espera pós-endodontia. Objetivo: Avaliar o efeito de diferentes cimentos endodônticos e dois tempos de espera pós-endodontia na resistência de união de pinos de fibra. Material e método: Setenta e dois dentes bovinos foram tratados endodonticamente e obturados usando três cimentos endodônticos: à base de eugenol, à base de resina epóxia ou à base de mineral trióxido agregado. Os espécimes foram armazenados a 37 ° C por 24 horas ou por 30 meses. Após, os canais radiculares foram preparados para cimentação dos pinos de fibra usando o RelyX U200. Foram realizados testes de push-out e análise de falhas. Os dados foram analisados por análise de variância bidirecional e com o teste t. Resultado: O cimento AH Plus obteve os maiores valores de resistência de união aos 30 meses pós-endodontia (11,26 Mpa) (p <0,05), no entanto, não houve diferença com o cimento Endofill no mesmo tempo. Os cimentos Endofill e MTA Fillapex não diferiram significativamente em seus efeitos, independentemente do tempo de espera pós-endodontia. Conclusão: O cimento endodôntico utilizado e o tempo de espera pós-endodontia afetam a resistência adesiva dos pinos de fibra. A adesão aumenta significativamente quando o pino de fibra é cimentado 30 meses após a obturação do canal radicular, enquanto a adesão é reduzida ao cimentar imediatamente após o tratamento do canal radicular, principalmente para cimentos endodônticos à base de eugenol.

Propriedades físico-químicas de cimentos endodônticos à base de óxido de zinco e eugenol / Physicochemical properties of zinc oxide and eugenol endodontic cements

Objetivo: Analisar as propriedades físico-químicas, escoamento, tempo de presa, solubilidade e alteração dimensional, dos cimentos endodônticos à base de óxido de zinco e eugenol: Endomethasone N, Endofill e Grosmann, de acordo com a especificação número 57 da American Dental Association. Métodos: Realizou-se os testes nas condições ambientais de 23° ± 2°C e 50 ± 5% de umidade relativa do ar, mantidas 48 h antes dos procedimentos, repetindo-se cinco vezes e obtendo-se uma média aritmética. Resultados: Os cimentos à base de óxido de zinco e eugenol analisados apresentaram tempo de endurecimento elevados, para o teste de escoamento, o Endofill e Grossman apresentaram-se acima do estabelecido. Em relação à solubilidade, o Endomethasone N esteve dentro do padrão e os demais cimentos excederam. Conclusão:Todos os cimentos analisados obtiveram valores diferentes do recomendado para alteração dimensional. Nenhum dos cimentos enquadrou-se completamente nas normatizações.

Aim: To analyze the physical-chemical properties, flow, setting time, solubility and dimensional change of endodontic cements, based on zinc oxide and eugenol: Endomethasone N, Endo fill, and Grosmann, according to specification number 57 of the American Dental Association. Methods: The tests were conducted under environmental conditions of 23° ± 2°C and 50 ± 5% relative air humidity, maintained for 48 h prior to the procedures, repeating five times and obtaining an arithmetic mean. Results: The cements based on zinc oxide and eugenol analyzed in this study showed high hardening times. For the flow test, Endo fill and Grossman were above the established parameters. Regarding solubility, Endomethasone N was within the standard, while the other cements exceeded this standard. Conclusion: All analyzed cements obtained values that proved to be different from those recommended for dimensional changes. None of the cements were completely compliant with the regulations.

The effectiveness of eugenol against cisplatin-induced ototoxicity / A eficácia do eugenol contra a ototoxicidade induzida pela cisplatina

Abstract Introduction: Ototoxicity refers to cellular damage or function impairment developing in the inner ear in association with any therapeutic agent or chemical substance, and still represents the principal side-effect restricting the use of cisplatin. Objective: The aim of this study was to perform a biochemical, functional and histopathological investigation of the potential protective effect of eugenol against cisplatin-induced ototoxicity. Methods: The study was performed with 24 female Sprague Dawley rats. Distortion product otoacoustic emissions tests were performed on all animals, which were randomized into four equal groups. A single intraperitoneal dose of 15 mg/kg cisplatin was administered to cisplatin group, while the eugenol group received 100 mg/kg eugenol intraperitoneal for five consecutive days. 100 mg/kg eugenol was administered to cisplatin + eugenol group for 5 days. On the third day, these rats were received a single dose of 15 mg/kg cisplatin. The control group was given 8 mL/kg/day intraperitoneal saline solution for five days. The distortion product otoacoustic emissions test was repeated 24 h after the final drug administration. All animals were sacrificed, and the cochleas were subsequently used for biochemical and histopathological examinations. Results: Cisplatin caused oxidative stress in the cochlea, impaired the cochlear structure and significantly reduced signal noise ratio levels. Administration of eugenol together with cisplatin reversed these effects and provided functional, biochemical and histopathological protection. Conclusion: The study findings represent the first indication in the literature that eugenol may protect against ototoxicity by raising levels of antioxidant enzymes and lowering those of oxidant parameters.

Resumo Introdução: A ototoxicidade refere-se ao dano celular ou comprometimento da função da orelha interna associado a qualquer agente terapêutico ou substância química e ainda representa o principal efeito colateral que restringe o uso da cisplatina. Objetivo: O objetivo deste estudo foi realizar uma investigação bioquímica, funcional e histopatológica do potencial efeito protetor do eugenol contra a ototoxicidade induzida pela cisplatina. Método: O estudo foi realizado com 24 ratos fêmeas Sprague Dawley. Testes de emissões otoacústicas por produto de distorção foram realizados em todos os animais, os quais foram randomizados em quatro grupos iguais. Uma única dose intraperitoneal de 15 mg/kg de cisplatina foi administrada ao grupo cisplatina, enquanto o grupo eugenol recebeu 100 mg/kg de eugenol intraperitoneal por cinco dias consecutivos. Foram administrados 100 mg/kg de eugenol ao grupo cisplatina + eugenol durante 5 dias. No terceiro dia, estes ratos receberam uma dose única de 15 mg/kg de cisplatina. O grupo controle recebeu 8 mL/kg/dia de solução salina intraperitoneal por cinco dias. O teste de emissões otoacústicas por produto de distorção foi repetido 24 horas após a administração final do medicamento. Todos os animais foram sacrificados e as cócleas foram posteriormente utilizadas para exames bioquímicos e histopatológicos. Resultados: A cisplatina causou estresse oxidativo na cóclea, prejudicou a estrutura coclear e reduziu significativamente os níveis da relação sinal/ruído. A administração de eugenol juntamente com a cisplatina reverteu esses efeitos e forneceu proteção funcional, bioquímica e histopatológica. Conclusão: Os achados do estudo representam a primeira indicação na literatura de que o eugenol pode proteger contra a ototoxicidade, eleva os níveis de enzimas antioxidantes e diminui os níveis dos parâmetros oxidantes.

Use of eugenol for anesthesia of lesser guitarfish Zapteryx brevirostris (Rhinobatidae) / Uso de eugenol para anestesia de raia-viola Zapteryx brevirostris (Rhinobatidae)

Abstract Anesthesia can be utilized as a non-lethal procedure to allow easy handling of teleosts and elasmobranchs in captivity or in the wild. For this, anesthetic protocols need to be established according to the species. The aim of this study was to determine the ideal concentration of eugenol for anesthesia of Zapteryx brevirostris. Four concentrations were tested: 21.25, 42.50, 85.00 and 170.00 mg L-1 (ratio of 1:5 with absolute ethanol). The perfect concentration of eugenol for this species was 85.0 mg L-1 , which enabled up to 300 seconds of work on the fish, without any response to handling.

Resumo A anestesia pode ser utilizada como um procedimento não letal que facilita o manejo de teleósteos e elasmobrânquios de cativeiro e de vida livre. Para isso, protocolos anestésicos devem ser estabelecidos de acordo com a espécie. O objetivo deste trabalho foi determinar a concentração ideal do eugenol para anestesiar Zapteryx brevirostris. Foram testadas as concentrações de 21,25; 42,50; 85,00 e 170,00 mg L-1 (1:5 de álcool absoluto). A concentração de 85,0 mg L-1 de eugenol foi a mais adequada para a espécie, permitindo uma janela de trabalho de até trezentos segundos com o animal não respondendo ao manejo.

Antibacterial activity and effects of aromatic derivatives on denture base polymerization

This study assessed the effect of copolymerization on the microbiota of the oral cavity. The plant extraction was converted into aromatic derivatives, which were added to methyl methacrylate monomer. Fourier transform infrared spectroscopy spectra showed no negative effects of these additives on the polymerization process. All the assayed derivatives displayed some degree of antibacterial activity.

Adaptação de Escherichia coli enterotoxigênica ao eugenol / Adaptation of enterotoxigenic Escherichia coli to eugenol

O objetivo do trabalho foi verificar a capacidade de adaptação de ETEC- ATCC 35401 ao antimicrobiano eugenol. Para isso, a Concentração Mínima Bactericida (CMB) de eugenol foi determinada e em seguida,as células de ETEC foram expostas a concentrações subletais de eugenol (CMB/4, CMB/8 e CMB/16). Posteriormente testadas frente a diferentes concentrações do composto (CMB/2; CMB; 1,2CMB; 1,4CMB; 1,6CMB; 1,8CMB e 2CMB), estas foram incubadas e plaqueadas em TSA (Ágar Triptona de Soja) empregando a técnica de microgotas. As células de ETEC foram classificadas como capazes de se adaptarem quando essas cresceram em placas após cultivo em presença do componente em concentração igual ou maior que a CMB do eugenol (1,0% (v/v)). As células de ETEC apresentaram a capacidade de adaptação por crescerem na CMB, após exposta a CMB/16. Os resultados demonstram a necessidade de obtenção da concentração de uso adequado de eugenol a fim de evitar a adaptação.

Direct biodegradation of eugenol to coniferyl aldehyde and other higher value-added products by Gibberella fujikuroi ZH-34

BACKGROUND: Eugenol is an economically favorable substrate for the microbial biotransformation of aromatic compounds. Coniferyl aldehyde is one kind of aromatic compound that is widely used in condiment and medical industries; it is also an important raw material for producing other valuable products such as vanillin and protocatechuic acid. However, in most eugenol biotransformation processes, only a trace amount of coniferyl aldehyde is detected, thus making these processes economically unattractive. As a result, an investigation of new strains with the capability of producing more coniferyl aldehyde from eugenol is required. RESULTS: We screened a novel strain of Gibberella fujikuroi, labeled as ZH-34, which was capable of transforming eugenol to coniferyl aldehyde. The metabolic pathway was analyzed by high-performance liquid chromatography­mass spectrometry and transformation kinetics. The culture medium and biotransformation conditions were optimized. At a 6 h time interval of eugenol fed-batch strategy, 3.76 ± 0.22 g/L coniferyl aldehyde was obtained, with the corresponding yield of 57.3%. CONCLUSIONS: This work improves the yield of coniferyl aldehyde with a biotechnological approach. Moreover, the fed-batch strategy offers possibility for controlling the target product and accumulating different metabolites

Chemical characterization of the essential oil from leaves of basil genotypes cultivated in different seasons / Caracterización química del aceite esencial de hojas genotipos de albahaca cultivados en diferentes estaciones del año

The aim of this study was to evaluate the concentration and chemical composition of the essential oil the leaves of basil cultivars and hybrids cultivated in different cropping seasons: dry season and rainy season. The variables evaluated were the content and composition of essential oils in the two seasons. The essential oil content ranged from 0.66% to 3.21% in the dry season and from 0.80% to 4.20% in the rainy season. The major compounds found among the genotypes were linalool, methyl chavicol, neral, geranial, eugenol, and methyl (E)- cinnamate, defining the formation of five groups in each season, classified in the following chemotypes: methyl chavicol (Group 1), citral (neral+geranial) (Group 2), methyl cinnamate (Group 3), linalool (Group 4), and intermediate linalool (Group 5). All the traits evaluated had heritability (h ) greater than 95% and high CVg/CVe ratio values. The cropping season affected the content and chemical compositions of basil essential oil.

El objetivo de este estudio fue evaluar la concentración y composición química del aceite esencial las hojas de cultivares de albahaca e híbridos cultivados en diferentes temporadas de cultivo: estación seca y estación lluviosa. Las variables evaluadas fueron la contenido y la composición de los aceites esenciales en las dos estaciones. La contenido de aceite esencial varió de 0.66% a 3.21% en la estación seca y de 0.80% a 4.20% en la estación lluviosa. Los principales compuestos encontrados entre los genotipos fueron linalool, metilchavicol, neral, geranial, eugenol y metil (E)-cinamato, definiendo la formación de cinco grupos en cada estación, clasificados en los siguientes quimiotipos: metil chavicol (Grupo 1), citral (neral + geranial) (Grupo 2), cinamato de metilo (Grupo 3), linalool (Grupo 4) y linalol intermedio (Grupo 5). Todos los rasgos evaluados mostraron una heredabilidad (h ) mayor que el 95% y altos valores de relación CVg/CVe. La temporada de cultivo afectó la contenido y las composiciones químicas del aceite esencial de albahaca.

Antinociceptive local activity of 4-allyl-1-hydroxy-2-methoxybenzene (eugenol) by the formalin test: an anti-inflammatory effect

Eugenol has been employed for decades as a condiment, an antimycotic, an antibacterial, an antiviral, and an antioxidant, and it is one of the natural analgesics most frequently utilized for pain and inflammation. Our objective was to determine the analgesic/anti-inflammatory effect of eugenol compared with diclofenac, naproxen, and tramadol using the formalin test. The formalin method was used in 6- to 10-week-old Wistar rats (weighing 250 g each) divided into six groups: saline (0.9%); formalin (5%); diclofenac (250 µg/kg); naproxen (400 µg/kg); tramadol (500 µg/kg), and eugenol (1,400 µg/kg), in the intraplantar part of the hind-end trunk of the rats, with n = 5 per group. Eugenol diminished 44.4% of nociceptive behavior in phase 1 and 48% in phase 2 (p ≤0.05 vs formalin). Eugenol was shown to be 1.14 times more effective than diclofenac, but 1.62 and 1.75 times less effective than naproxen and tramadol, respectively, in phase 1 and 1.45 times less effective than diclofenac and naproxen and 1.66 less effective than tramadol in phase 2 (p ≤0.05). These data suggest that eugenol possesses moderate activity in the acute pain phase and greater activity in inflammatory-type pain, and both effects are comparable to those produced by diclofenac and are less than the effects produced by naproxen and tramadol in the formalin test