Difference of Chemical Bonds Between UDMA Bonding Agents with Ethanol Solvent and Acetone Solvent on Dentin Collagen

ABSTRACT Objective: To investigate the difference of chemical bonds between urethane dimethacrylate (UDMA) bonding agents with ethanol solvent and acetone solvent on dentin collagen. Material and Methods: This experimental comparison study used three groups: G1 (Control): UDMA and collagen; G2: UDMA, collagen and ethanol; and G3: UDMA, collagen and acetone. The groups were then pelleted and analysed with FTIR, then the peak value of carbonyl absorption band from each study group was calculated. The result of FTIR analysis and the peak of carbonyl absorption band (P) was calculated using the formula: P = (BC / AB) X 100; AB. BC is measured in centimeters. The study of chemical bond differences between ethanol-solvent UDMA agents compared with acetone-solvent on dentin collagen resulted in a graph of peak of carbonyl absorption bands of UDMA and dentin collagen groups. To determine the chemical bonds of UDMA from the top of the carbonyl ester absorption bands with wavenumber absorption in range 1700-1750 cm-1, the decreasing peak of the carbonyl absorption bands is assumed as more chemical bonds that formed. Data were analysed using Anova one way and Tukey HSD test. Results: There were significant differences between the three study groups (p<0.05). Conclusion: UDMA bonding agents' chemical bonds with acetone solvent are much higher than the chemical bonds between UDMA bonding agents with ethanol solvent on dentin collagen.

Efecto del extracto hidroalcohólico de la planta Lampaya medicinalis Phil. (Verbenaceae) sobre la esteatosis inducida in vitro en hepatocitos humanos / Effect of the hydroalcoholic extract of the plant Lampaya medicinalis Phil. (Verbenaceae) on in vitro-induced steatosis in human hepatocytes

INTRODUCCIÓN: La enfermedad del hígado graso no alcohólico (EHGNA) es la forma más común de enfermedad hepática. A nivel celular se caracteriza por la acumulación de triglicéridos (TG) en forma de gotas lipídicas (GL) dando lugar a esteatosis e inflamación. Entre los factores relevantes para la síntesis de TG se encuentran las enzimas DGAT1/2 que catalizan la etapa final de la síntesis de TG, y la proteína FABP4 que transporta lípidos intracelulares y se expresa en modelos de enfermedad hepática dependiente de obesidad. Por otra parte, TNF-α es una reconocida citoquina involucrada en el proceso inflamatorio en la EHGNA. La medicina popular del norte de Chile ha utilizado la planta Lampaya medicinalis Phil. (Verbenaceae) para el tratamiento de algunas enfermedades inflamatorias. OBJETIVO: Evaluar el efecto de un extracto hidroalcóholico de lampaya (EHL) sobre la esteatosis y expresión de marcadores de inflamación en hepatocitos tratados con ácidos grasos. Diseño experimental: Estudio in vitro en cultivos de la línea celular humana HepG2 tratadas con ácido oleico (AO) y ácido palmítico (AP). MÉTODOS: Se incubó hepatocitos HepG2 con AO/AP por 24 horas en presencia o no de EHL. Se evaluó la presencia de GL y el contenido de TG intracelulares por Oil Red O y Nile Red, respectivamente. La expresión de DGAT1/2, FABP4 y TNF-α fue evaluada por qPCR. RESULTADOS: Los hepatocitos tratados con AO/AP mostraron un aumento en las GL y TG, así como una mayor expresión de DGAT2 en comparación al control. El cotratamiento con EHL revirtió los efectos inducidos por AO/AP. CONCLUSIONES: EHL revierte el incremento en las GL, TG y en la expresión de DGAT2 inducido por AO/AP en células HepG2. Estos hallazgos sugieren un efecto hepatoprotector de la Lampaya contra la esteatosis, y apoyarían su uso complementario en el tratamiento de patologías con componente inflamatorio como la EHGNA.

Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease. At the cellular level, it is characterized by the accumulation of triglycerides (TG) in the form of lipid droplets (LD), which leads to steatosis and inflammation. Among relevant factors for TG synthesis are the enzymes DGAT1/2 catalyzing the final stage of TG synthesis, and the protein FABP4 which transports intracellular lipids and is expressed in cell models of obesity-dependent liver disease. Additionally, TNF-α is a cytokine involved in the inflammatory process associated to NAFDL. Lampaya medicinalis Phil. (Verbenaceae) is a plant used in folk medicine in northern Chile to treat some inflammatory diseases. OBJECTIVE: To evaluate the effect of the hydroalcoholic extract of lampaya (HEL) on steatosis and the expression of inflammatory markers in hepatocytes treated with fatty acids. Study design: In vitro study in cultures of the human HepG2 cell line treated with oleic acid (OA) and palmitic acid (PA). METHODS: HepG2 hepatocytes were incubated with OA/PA for 24 hours in the presence and absence of HEL. The formation of LD and the accumulation of intracellular TG were assessed by Oil Red O and Nile Red, respectively. The expression of DGAT1/2, FABP4 and TNF-α was assessed by qPCR. RESULTS: The treatment with OA/PA increased the levels of LD and TG as well as the expression of DGAT2 in HepG2 hepatocytes compared to control cells. HEL cotreatment counteracted OA/PA-induced effects. CONCLUSIONS: HEL prevents the increase in LD and TG levels and DGAT2 expression induced by OA/PA in HepG2 cells. These findings suggest that lampaya may have a protective effect against hepatic steatosis, which would support its complementary use in the treatment of pathologies associated with inflammation, such as NAFLD.

Influence of ethanol on dentin roughness, surface free energy, and interaction between AH Plus and root dentin

Abstract This study aimed to evaluate the influence of different ethanol concentrations on dentin roughness, surface free energy, and contact angle between AH Plus and the root canal dentin. One hundred human maxillary anterior teeth were split longitudinally and 200 dentin specimens were polished to make the surface flatter and smoother. An acrylic bar was positioned between two dentin specimens and impression material was added to create a block, simulating an instrumented root canal space. Specimens were removed from the mold and cleaned in an ultrasonic bath for 10 min. Thereafter, dentin specimens were divided into four groups (n = 50) according to the drying methods used: a) wet: vacuum only, b) paper points: vacuum + absorbent paper points, c) 70% alcohol: 70% alcohol (1 min) + vacuum + absorbent paper points, and d) 100% alcohol: 100% alcohol (1 min) + vacuum + absorbent paper points. A rugosimeter and a goniometer were used to verify the roughness (Ra) and to measure the surface free energy and the contact angle between the AH Plus sealer and the root canal dentin. ANOVA and Tukey tests (α = 0.05) were used for statistical analysis. The 70% and 100% ethanol groups showed significantly decreased roughness as well as increased surface free energy in the root canal dentin when compared to the wet and paper point groups. In addition, ethanol significantly reduced the contact angle between the AH Plus sealer and the root canal dentin. Ethanol solutions (70% and 100%) provide better wettability of AH Plus sealer on dentin surfaces.

In vitro effects of alcohol-containing mouthwashes on human enamel and restorative materials

Abstract The objective of this study was to evaluate the in vitro effects, including surface morphological characteristics and chemical elemental properties, of different mouthwash formulations on enamel and dental restorative materials, simulating up to 6 months of daily use. Human enamel samples, hydroxyapatite, composite resin, and ceramic surfaces were exposed to 3 different mouthwashes according to label directions — Listerine® Cool Mint®, Listerine® Total Care, and Listerine® Whitening — versus control (hydroalcohol solution) to simulate daily use for up to 6 months. The samples were analyzed using scanning electron microscopy (SEM), infrared spectrophotometry (µ-Fourier transform infrared microscopy), energy-dispersive X-ray (EDX) spectroscopy, and color analysis before and after exposure. No relevant changes were observed in the morphological characteristics of the surfaces using SEM techniques. The physical and chemical aspects of the enamel surfaces were evaluated using mid-infrared spectroscopy, and EDX fluorescence was used to evaluate the elemental aspects of each surface. There was no variation in the relative concentrations of calcium and phosphorus in enamel, silicon and barium in composite resin, and silicon and aluminum in the ceramic material before and after treatment. No relevant changes were detected in the biochemical and color properties of any specimen, except with Listerine® Whitening mouthwash, which demonstrated a whitening effect on enamel surfaces. Long-term exposure to low pH, alcohol-containing, and peroxide-containing mouthwash formulations caused no ultra-structural or chemical elemental changes in human enamel or dental restorative materials in vitro.

Influence of ethanol-wet dentin, adhesive mode of application, and aging on bond strength of universal adhesive

Abstract The aim of this study was to evaluate the effect of ethanol on the bond longevity of a universal adhesive system to bovine dentin, under different modes of adhesive application and artificial aging. Bovine dentin was exposed, and the smear layer was standardized by sandpaper polishing. Specimens were randomly divided into 2 groups: ethanol (E) and non-ethanol (N). Groups were subdivided according to adhesive mode of application into etch-and-rinse (Er) and self-etching (S). Resin blocks were built onto the treated surface, and the specimens were stored in deionized water at 37°C for 48 h. Half of the specimens (n = 10) were subjected to thermomechanical aging (A for aged and Na for non-aged). Resin/dentin beams were obtained and subjected to microtensile test in a universal testing machine. Data were analyzed using a three-way ANOVA and Tukey's tests (α = 5%). There was interaction among the three factors (p=0.0003). The use of ethanol resulted in higher values, except for the Er and Na groups (E_Er_Na = N_Er_Na). The mode of application was similar, except for the N and A groups (N_S_A > N_Er_A). For the A groups, the values were lower, except in the cases using ethanol, in which the results were not affected. The study concluded that the use of ethanol resulted in higher microtensile bond strength values, even after aging. The mode of adhesive application did not influence the results.

Influence of a hydrophobic monomer on the physical and mechanical properties of experimental surface sealants

Abstract This study evaluated the effect of adding the hydrophobic monomer 1,12 dodecanediol dimethacrylate (DDDMA) to experimental sealants with and without thermocycling on degree of conversion (DC), water sorption (WS), water solubility (WSB), color stability (ΔE), and micro-shear bond strength (μSBS). Five experimental and one commercially available sealant (Bisco - BIS) were tested. The experimental sealants were formulated by mixing different percentages of DDDMA monomers and urethane dimethacrylate (UDMA). The photoinitiator system was composed by camphorquinone (CQ) and tertiary amine 4-ethyl benzoate dimetilamiono (EDBA). Ethanol was used as a solvent. The experimental groups were named sequentially according to the monomeric content (DDDMA/UDMA): S40/40 (40/40), S50/30 (50/30), S60/20 (60/20), S70/10 (70/10) and S80/0 (80/0). Data were analyzed separately by one-way ANOVA, followed by Tukey's test (p<0.05). The values of DC ranged from 94.59% (S40/40) to 54.02% (S80/10). BIS showed the highest WS value (p<0.05) and S40/40, S50/30, S60/20 and S80/0 showed the lowest WS values of all tested sealants. WSB values ranged from 7.88 µg/mm3 (BIS) to 13.27 µg/mm3 (S70/10). The highest ΔE value was 11.05±2.88 for BIS and the highest μSBS value was found for S60/20. No significant difference was observed in bond strength between sealants and bovine enamel after thermocycling. Adding DDDMA to the composition of surface sealants can improve its performance, once the monomer increased the degree of conversion and the color stability.

Susceptibilidad de Cepas de Candida Oral a Extracto Etanólico del Propóleo Chileno de Olmué / Susceptibility of Oral Candida Strains to Ethanol Extract of Chilean Propolis of Olmué

RESUMEN: La infección por Candida albicans en la mucosa oral es conocida como Candidiasis oral (CO) y se diagnostica por el reconocimiento de cambios clínicos y la presencia de pseudohifas, hifas o levaduras en muestras obtenidas por citología exfoliativa o biopsia. Los agentes farmacológicos tópicos clásicos para el tratamiento de CO son Nistatina y Miconazol. Sin embargo, a pesar de las distintas terapias contra CO, existen formas de Candida resistentes al tratamiento convencional. El objetivo de este estudio fue determinar la susceptibilidad in vitro de Candida spp. a un extracto etanólico de propóleo de Olmué. Se realizó un estudio experimental descriptivo in vitro en donde se evaluó el efecto que presenta el uso de extracto etanólico de propóleo como antifúngico sobre cepas de Candida spp. obtenidas de la cavidad oral (mucosa palatina) de 31 individuos, con candidiasis oral diagnosticados con estomatitis subprotésica. El propóleo chileno utilizado fue obtenido de la zona geográfica de Olmué, quinta región. Se encontró que el 100 % de las muestras en rangos de concentración de propóleo de 0,1 µg/mL y 1,6 µg/mL presentaron un grado de inhibición en el crecimiento de Candida Oral y por otra parte el extracto etanólico de propóleo que generó inhibición en la mayor cantidad de muestras fue al 0,4 µg/mL (41,94 % de las muestras) y en segundo lugar la concentración al 0,2 µg/mL (35 % de las muestras). Se concluyó que el extracto etanólico de propóleo chileno obtenido de la zona de Olmué presenta la capacidad de inhibir el crecimiento de Candida spp. en agar Sabouraud in vitro de forma dosis dependiente.

ABSTRACT: Fungal (or yeast) infections; mycoses, occurring in the oral mucous membranes, of Candida species (mostly C. albicans, a normal component of the oral microbiota), also known as oral thrush or oral candidiasis (OC), can be diagnosed via the recognition of clinical changes and the presence of pseudohyphae, hyphae or yeasts in samples obtained by exfoliative cytology and/or biopsy. Topical pharmacological preparations and drugs such as Nystatin and Miconazole are used in the treatment of CO. However, there are forms of Candida with resistance to such conventional treatment approaches. Therefore, the aim of this study is to determine the in vitro susceptibility of Candida spp.; an ethanolic extract of propolis from Olmué. Hence, an experimental in vitro descriptive study was carried out in which the effect of an ethanolic extract of propolis used as antifungal on strains of Candida spp. obtained from the oral cavity (palatine mucosa) of 31 individuals, diagnosed with OC (subdenture stomatitis) is determined. Natural propolis was obtained from the Olmué area, in the 5th region of Chile. It was found that 100 % of the samples with propolis concentration ranging from 0.1 mg / mL to 1.6 mg / mL presented a degree of inhibition in the growth of OC. On the other hand, the ethanolic extract of propolis that generated inhibition in the largestnumber of samples was 0.4 mg / mL (41.94 % of the samples) followed by the concentration of 0.2 mg / mL (35 % of the samples). Therefore, it can be concluded that the ethanolic extract of Chilean propolis obtained from the Olmué area has the ability to inhibit the growth of Candida spp. in vitro in a dosage-dependent manner.

Screening of plant extracts and fractions on Aedes aegypti larvae found in the state of Mato Grosso do Sul (linnaeus, 1762) (culicidae)

ABSTRACT The constant use of chemical insecticides for Aedes aegypti control has caused resistance in the mosquito populations. Thus, the objective of this study was to analyze the larvicidal potential of extracts and fractions of plants on A. aegypti larvae. The analysis included sixty one extracts and twenty five fractions of fifty botanical species at concentrations of 0.25; 0.12; 0.06 to 0.03 mg mL-1; 4 replications and one negative control of dechlorinate water and 1% DMSO; and a positive control with rotenone. The toxicity index in descending order with LC50 for the most active of the extracts selected were ethanol extract of Ormosea arborea (0.111 mg mL-1) seeds and ethanol extracts of leaves such as Piper hispidum (0.169 mg mL-1), Solanum variabile (0.188 mg mL-1), O. arborea (0.238 mg mL-1), Turnera umifolia (0.242 mg mL-1) and Piper hispidum (0.567 mg mL-1). For plant fractions, the most active were chloroform (0.192 mg mL-1) and hexane (0.342 mg mL-1) P. aduncum leaves, hexane fraction (0.415 mg mL-1) and methanol extract (0.625 mg mL-1) of Spermacocea latifolia leaves. Regarding the extract of T. umifolia single species, there is no bibliographic report on their degree of efficiency as an insecticide.

Evaluation of cytogenotoxicity, antioxidant and hypoglycemiant activities of isolate compounds from Mansoa hirsuta D.C. (Bignoniaceae)

ABSTRACT Mansoa hirsuta (Bignoniaceae) is a native plant from caatinga in Brazilian semiarid. This plant has been locally used as antimicrobial and hypoglycemiant agents, but their action mechanisms and toxicity remain largely unknown. Therefore, we evaluated the composition and antioxidant, cytoprotective and hypoglycemiant effects of raw extract, fractions and compounds from leaves of M. hirsuta. The cytogenotoxic effects of ursolic and oleanolic acids, the main phytotherapic components of this plant, were assessed. The raw extract and fractions presented steroids, saponins, flavonols, flavanonols, flavanones, xanthones, phenols, tannins, anthocyanins, anthocyanidins and flavonoids. The ethyl acetate fraction inhibited efficiently the cascade of lipid peroxidation while the hydroalcoholic fraction was richer in total phenols and more efficient in capturing 2,2-diphenyl-1-picrylhydrazyl (·DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS·+) radicals. The isolated fraction of M. hirsuta also inhibited the α-amylase activity. Cytotoxic effects were absent in both raw extract and fractions while ursolic+oleanolic acids were efficient in protecting cells after exposure to hydrogen peroxide. Moreover, this mixture of acid shad no significant interference on the mitotic index and frequency of nuclear and/or chromosomal abnormalities in Allium cepa test. Therefore, M. hirsuta represents a potential source of phytochemicals against inflammatory and oxidative pathologies, including diabetes.

Molecular and morphological surface analysis: effect of filling pastes and cleaning agents on root dentin

Abstract The quality of the dentin root is the most important factor for restoration resin sealing and drives the outcome of endodontic treatment. Objective This study evaluated the effect of different filling pastes and cleaning agents on the root dentin of primary teeth using Fourier-transformed Raman spectroscopy (FT-Raman), micro energy-dispersive X-ray fluorescence (µ-EDXRF) and scanning electron microscopic (SEM) analysis. Material and Methods Eighty roots of primary teeth were endodontically prepared and distributed into 4 groups and filled according to the following filling pastes: Control-no filling (CP), Calen®+zinc oxide (CZ), Calcipex II® (CII), Vitapex® (V). After seven days, filling paste groups were distributed to 4 subgroups according to cleaning agents (n=5): Control-no cleaning (C), Ethanol (E), Tergenform® (T), 35% Phosphoric acid (PA). Then, the roots were sectioned and the dentin root sections were internally evaluated by FT-Raman, µ-EDXRF and SEM. Data was submitted to two-way ANOVA and Tukey tests (α=0.05). Results Regarding filling pastes, there was no significant difference in organic content. CP provided the lowest calcium values and, calcium/phosphoric ratio (Ca/P), and the highest phosphoric values. For cleaning agents there was no difference in organic content when compared to the C; however, T showed significantly higher calcium and Ca/P than PA. All groups showed similar results for phosphorus. The dentin smear layer was present after use of the cleaning agents, except PA. Conclusion The filling pastes changed the inorganic content, however they did not change the organic content. Cleaning agents did not alter the inorganic and organic content. PA cleaned and opened dentin tubules.

Chlorhexidine with or without alcohol against biofilm formation: efficacy, adverse events and taste preference

Abstract In recent years, different chlorhexidine formulations have been tested, including an alcohol-free alternative, but the effect of this solution on early biofilm formation is not clear. A crossover, randomized, double-blind clinical trial was conducted to evaluate the effect of two chlorhexidine solutions against supra- and subgingival biofilm formation (NCT#02656251). Thirty-five participants were randomized and asked to rinse twice daily with 15 ml of an alcohol-containing 0.12% chlorhexidine solution, an alcohol-free 0.12% chlorhexidine solution, or placebo. The study was conducted in three experimental periods of 4 days each, with a 10-day washout between the periods. All the experimental periods followed the same protocol, except that the solutions were switched. Biofilm distribution was evaluated every 24 hours by the Plaque-Free Zone Index, during 96 hours. Adverse events were self-reported and sensory evaluation was performed using a hedonic scale. Compared to the placebo, the chlorhexidine solutions resulted in a significantly higher number of surfaces free of plaque over 96 hours (p < 0.01), and were able to prevent subgingival biofilm formation (p < 0.01). The alcohol-free chlorhexidine solution was associated with a lower incidence of adverse events, compared with alcohol-containing chlorhexidine (p < 0.05); it also received better sensory evaluation and acceptance by trial participants, compared with the alcohol-containing chlorhexidine (p = 0.007), and had a similar inhibitory effect on the formation of supra- and subgingival biofilms.

Effect of different presentations of resveratrol on cell proliferation and epitelial thickness of the oral mucosa of wistar rats

Introduction: Grape is one of the most important fruit crops across the world and can be consumed in different ways. There has been a growing interest in the role of antioxidants such as resveratrol, which can be found in grape skin, in oral and dental tissues. Thus, the objective of this study was to analyze the effect of different presentations of resveratrol on cell proliferation and epithelial thickness of the oral mucosa of Wistar rats. Methods: Fifty male Wistar rats were randomly divided into five groups: water/control, red wine, grape juice, 12% alcoholic solution/ethanol and aqueous solution of resveratrol. Samples of palatal and tongue mucosa were collected for a histomorphometric analysis using hematoxylin-eosin staining and the argyrophilic nucleolar organizer region (AgNOR) technique for quantification of cell proliferation. Results: As to epithelial thickness, both the tongue and the palate showed a statistically significant difference between the control group and the other groups, with greater decrease in the resveratrol and the wine groups. In the suprabasal layer of both the tongue and the palate epithelium, red wine reduced the rate of cell proliferation, while ethanol increased it. In the basal layer of the tongue epithelium, there was a statistically significant difference between the control, the grape juice and the resveratrol groups and the ethanol group, with increased cell proliferation in the ethanol group. Conclusions: Wine does not interfere in the physiological renewal of the basal layer of the buccal epithelium and exerts a protective action by reducing the cell proliferation rate of the suprabasal layer (AU)

Efeito de adição de rodamina B e fluoresceína sódica a sistemas adesivos não simplificados: aspectos fotofísicos e físico-químicos / Effect of addition of rhodamine B and fluorescein to conventional etch-and-rinse and self-etching adhesive systems: photophysical and physical-chemical aspects

A adição de corantes fluorescentes a adesivos odontológicos possibilita a investigação da distribuição espacial desses materiais na interface dente-restauração, utilizando-se a microscopia confocal de varredura a laser (MCVL). A literatura indica falta de padronização na aplicação de agentes fluorescentes com tal finalidade. Esse estudo sistematizou estratégias para a adição de rodamina B (RB) e fluoresceína sódica (FS) a um sistema adesivo convencional de três passos, Adper Scotchbond Multi-Purpose (MP), e um autocondicionante de dois passos, Clearfil SE Bond (SE), considerados "padrão-ouro" na Odontologia. Os objetivos principais foram (a) determinar a menor faixa de concentrações de RB e FS necessária para produzir imagens satisfatórias da interface dentina-adesivo e (b) avaliar o efeito da adição desses corantes sobre algumas propriedades das resinas. Os adesivos foram marcados com RB ou FS em concentrações decrescentes (0,5, 0,1, 0,02 e 0,004 mg/mL) por meio de um método de dispersão semidireto. O comportamento fotofísico/ fluorescente dos adesivos marcados foi investigado por espectroscopia de fotoluminescência e MCVL. Paralelamente, avaliaram-se os adesivos quanto ao grau de conversão (GC) e ao ângulo de contato (AC). Tanto os resultados de GC como os de AC foram submetidos à análise de variância com dois fatores (adesivo e tratamento) com α = 0,05, seguida de teste post-hoc de Tukey. Os máximos comprimentos de onda de emissão e de excitação da RB e da FS foram influenciados pelo meio polimérico e pela concentração de corante de modo geral. A MCVL preliminar de amostras de adesivo polimerizado, realizada sob condições experimentais padronizadas, mostrou que o comportamento fluorescente da RB em MP e SE foi muito semelhante na mesma concentração de corante, mas o mesmo não pôde ser dito do comportamento da FS, que foi notavelmente inferior no adesivo autocondicionante, SE, na concentração mais alta. Em dentina, os adesivos preparados com RB nas concentrações-alvo de 0,1 e 0,02 mg/mL apresentaram fluorescência ótima; já aqueles preparados com 0,004 mg/mL produziram fraco sinal. Adesivos preparados com FS a 0,5 mg/mL apresentaram ótima fluorescência na interface de adesão, enquanto que concentração menor desse corante não produziu sinal suficiente. Padrões morfológicos aparentemente atípicos foram observados na interface de adesão, quando da associação do adesivo SE com o corante FS. A adição de RB e FS nas quatro concentrações indicadas aos adesivos MP e SE não afetou o GC nem o AC em comparação com os grupos de controle correspondentes. Em suma, a RB mostra-se um corante mais versátil que a FS na avaliação morfológica das interfaces dentina-MP e dentina-SE via MCVL. A menor faixa de concentrações de RB nos adesivos MP e SE, na qual é possível produzir imagens satisfatórias das interfaces, situa-se entre 0,1­0,02 mg/mL. Já o corante FS deve ser adicionado a esses adesivos a pelo menos 0,5 mg/mL para produzir níveis de fluorescência satisfatórios na interface de adesão. A não ocorrência de efeitos deletérios sobre a polimerização e a molhabilidade das resinas estabelece uma margem de segurança para a incorporação desses agentes fluorescentes (em concentração ≤ 0,5 mg/mL) nesses sistemas monoméricos.(AU)

The addition of fluorescent dyes to dental adhesives makes it possible to investigate the spatial distribution of such resin-based materials in the tooth-restoration interface, using confocal laser scanning microscopy (CLSM). Literature indicates a lack of standardization on the application of fluorescent agents for this purpose. This work presents strategies for adding rhodamine B (RB) and fluorescein sodium salt (FS) to a three-step etch-and-rinse adhesive system, Adper Scotchbond Multi-Purpose (MP), and a two-step self-etching one, Clearfil SE Bond (SE), both regarded as "gold standard" in restorative dentistry. The main objectives were (a) to determine the lowest range of RB and FS concentrations required to produce suitable images of the dentin-adhesive interface via CLSM and (b) to investigate potential effects of addition of these dyes on some resin properties. The adhesives were labeled with RB or FS at decreasing concentrations (0.5, 0.1, 0.02 and 0.004 mg/mL) by means of a semi-direct dispersion method. The photophysical/fluorescent behavior of the labeled resins was investigated by photoluminescence spectroscopy and by CLSM. The adhesives were also investigated with regards to the degree of conversion (DC) and contact angle (CA). A two-way ANOVA of "adhesive" and "treatment" was conducted on DC and CA separately, followed by Tukey's test. The maximum emission and excitation wavelengths of RB and FS were influenced by the host polymer and the dye concentration in general. The preliminary CLSM of cured adhesive samples, performed with standardized settings, showed that the fluorescent behavior of RB in MP and SE was very similar in the same dye concentration, unlike the behavior of FS, which was lower in the self-etching adhesive for the highest dye concentration. In dentin, the adhesives prepared with RB at the target concentrations of 0.1 and 0.02 mg/mL presented optimal fluorescence; those with 0.004 mg/mL produced poor signal. Adhesives prepared with FS at 0.5 mg/mL presented optimal fluorescence at the bonding interface, whereas lower concentrations of FS did not produce sufficient signal. Atypical morphological features were observed at the bonding interface, when adhesive SE was used with FS. The addition of RB and FS at the four decreasing concentrations to adhesives MP and SE did not affect DC or CA compared to the corresponding controls. In short, RB is more versatile than FS for the morphological characterization of dentin-MP and dentin-SE interfaces via MCVL. The lowest range of RB concentrations in adhesives MP and SE that can produce suitable images of the bonding interface lies between 0.1­0.02 mg/mL. The dye FS should be added to these adhesives at 0.5 mg/mL at least to produce satisfactory fluorescence levels at the bonding interface. Since negative effects on polymerization and wettability of the resins were not observed, the use of RB and FS (in concentration ≤ 0.5 mg/mL) together with MP and SE should be reliable in terms of resin properties.(AU)

Actividad antibacteriana in vitro de Curcuma longa (Zingiberaceae) frente a bacterias nosocomiales en Montería, Colombia / In vitro antibacterial activity of Curcuma longa (Zingiberaceae) against nosocomial bacteria in Montería, Colombia

ResumenLa resistencia bacteriana es un problema de salud creciente a nivel mundial que genera graves impactos económicos y sociales, comprometiendo la acción terapéutica de los antibióticos actuales. Por ello, la búsqueda de nuevos compuestos con propiedades antimicrobianas se hace más relevante en los estudios modernos, en especial frente a bacterias de interés clínico. En el presente estudio se evaluó la actividad antibacteriana in vitro del extracto etanólico y el aceite esencial de Curcuma longa L (Zingiberaceae), contra bacterias nosocomiales utilizando el método de microdilución. Se utilizaron cepas de Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus sp., Salmonella sp. y Bacillus sp., aisladas de infecciones nosocomiales en un centro hospitalario de la ciudad de Montería y cepas de referencia de S. aureus ATCC 43300, S. aureus ATCC 29213, S. aureus ATCC 25923, P. aeruginosa ATCC 27853, E. coli ATCC 25922 y K. pneumoniae ATCC 700603. El perfil antibacterial del extracto etanólico fue más evidente a las concentraciones más altas (1 000 ppm), obteniendo porcentajes de reducción significativos de más del 50 % frente a K. pneumoniae ATCC 700603 y un aislado clínico de E. coli, mientras que, frente al aislado clínico del género Bacillus fue más activo el aceite esencial. Para el resto de los microorganismos los porcentajes de reducción obtenidos a una concentración de 1 000 ppm variaron entre 17 y 42 % con el extracto etanólico y entre 8 y 43 % con el aceite esencial. A concentraciones de 100 y 500 ppm la actividad antibacteriana de los extractos fue menor. Los resultados obtenidos indican que el extracto etanólico y el aceite esencial de los rizomas de C. longa poseen compuestos activos con propiedades antibacterianas que podrían emplearse en investigaciones futuras, como una alternativa terapéutica para el tratamiento de infecciones producidas por patógenos nosocomiales.

Abstract:Bacterial resistance is a growing health problem worldwide that has serious economic and social impacts, compromising public health, and the therapeutic action of current antibiotics. Therefore, the search for new compounds with antimicrobial properties is relevant in modern studies, particularly against bacteria of clinical interest. In the present study, in vitro antibacterial activity of the ethanol extract and essential oil of Curcuma longa (Zingiberaceae) was evaluated against nosocomial bacteria, using the microdilution method. Escherichia coli strains, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus sp. were used, Salmonella sp. and Bacillus sp., isolated from nosocomial infections in a hospital in the city of Monteria and reference strains of S. aureus ATCC 43300, S. aureus ATCC 29213, S. aureus ATCC 25923, P. aeruginosa ATCC 27853, E. coli ATCC 25922 and K. pneumonia ATCC 700603. The ethanol extract antibacterial profile was more efficient at higher concentrations (1 000 ppm), obtaining significant percentages of reduction of more than 50 % against K. pneumoniae ATCC 700603 and a clinical isolate of E. coli; while compared to Bacillus clinical isolate, was more active than the essential oil. For the rest of microorganisms, the reduction percentages obtained at a concentration of 1 000 ppm varied between 17 and 42 % with ethanolic extract, and 8 to 43 % with essential oil. At concentrations of 100 and 500 ppm antibacterial activity of the extracts was lower. The results indicated that the ethanolic extract and essential oil of C. longa rhizomes have active compounds with antibacterial properties that could be used in future research as a therapeutic alternative for the treatment of infections caused by nosocomial pathogens. Rev. Biol. Trop. 64 (3): 1201-1208. Epub 2016 September 01.

Ethanol-wet bonding technique: 18-month clinical evaluation / Técnica de adhesión húmeda en etanol: evaluación clínica a los 18 meses

The objective of this randomized clinical trial was to evaluate the clinical performance up to 18 months of restorations placed using ethanol-wet bonding technique (EWBT) compared with the three-step etch-and-rinse (TSER) and one-step self-etching (OSSE) approaches. Ninety-three non-carious cervical lesions (31 for each group) were restored by one experienced operator in 17 patients under relatively dry conditions using gingival retraction cord, cotton rolls and saliva ejector. Each adhesive system was randomly allocated to one of randomized cervical lesions until the three groups were present in the same subject in equal amounts. The restorations were evaluated at baseline, 6, 12 and 18 months by two blinded and calibrated examiners using the modified US Public Health Service guidelines (USPHS) for the following outcomes: retention (kappa= 1.00), staining and marginal adaptation (kappa=0.81) and analyzed by Fisher's exact and Kruskal-Wallis tests, respectively. No significant differences were observed among groups after 18 months for any of the assessed criteria (p>0.05). The intra-group analysis performed by Cochran's test (for retention) and Wilcoxon test (for marginal adaptation/staining) revealed significant differences between the time intervals baseline/18 months in marginal adaptation (p= 0.0117) and retention (p= 0.0101) for OSSE and in marginal staining for TSER (0.0051) and EWBT (p= 0.0277) groups. The survival analysis for retention criteria and the overall clinical success were performed using a log-rank test and did not show significant differences among groups (p> 0.05). All three adhesives protocols presented similar clinical performance up to 18 months.

El propósito de este ensayo clínico aleatorizado fue evaluar, durante 18 meses, el éxito clínico de las restauraciones realizadas por la técnica de adhesión húmeda en etanol (TAHE) en comparación con las técnicas de grabado independiente de tres pasos (GTP) y de autograbado de un paso (AUP). Un solo operador, especializado en odontología restaurativa, realizó sobre 17 pacientes 93 restauraciones en lesiones cervicales no cariosas (31 para cada grupo). Los diferentes protocolos adhesivos se distribuyeron aleatoriamente en las lesiones cervicales hasta que los tres grupos estuviesen presentes en el mismo paciente y en cantidades iguales. Las restauraciones fueron evaluadas a los 6, 12 y 18 meses, por dos examinadores calibrados y que no participaron del procedimiento restaurador. Las directrices modificadas del Servicio de Salud Pública de Estados Unidos (SSPEU) fueron la base para las evaluaciones de las siguientes variables: retención (kappa= 1,00), adaptación y decoloración marginal (kappa= 0,81). Estas variables fueron analizadas mediante la prueba exacta de Fisher y Kruskal-Wallis, respectivamente. No se observó diferencia significativa entre los grupos después de 18 meses en las tres variables evaluadas (p >0,05). El análisis intra-grupo hecho por la prueba de Cochran (para la retención) y la prueba de Wilcoxon (para la adaptación y decoloración marginal) revelaron diferencias significativas entre los intervalos de tiempo de la línea de base / 18 meses para la retención (p= 0,0101) y adaptación marginal en el grupo AUP (p= 0,0117), y para la decoloración marginal en los grupos GTP (p= 0,0051) e TAHE (p= 0,0277). El análisis de supervivencia para la retención, así como la comparación del éxito clínico de los protocolos adhesivos, fueron realizados con la prueba de log-rank, y no hubo diferencias significativas (p >0,05) entre los grupos. No hubo diferencia en el éxito clínico de los tres protocolos adhesivos después de 18 meses.

Antimicrobial effect of ethanol extract of leaf neem (Azadirachta indica A. Juss) on Listeria monocytogenes

Listeria monocytogenes es un patógeno causante de enfermedades alimentarias. En la búsqueda de controlar su propagación utilizando sustancias naturales se planteó el objetivo de mostrar si el extracto etanólico foliar de neem (Azadirachta Indica A. Juss.) tiene efecto antimicrobiano sobre L. monocytogenes ICTA-12446. El extracto se obtuvo a partir de hojas de neem sometidas a secado por 8 días, se redujeron de tamaño mecánicamente, se sometieron a maceración en frío por 3 días usando etanol 96% en recipientes ámbar, se filtró y concentró en rota evaporador. Se estandarizó el concentrado con dimetilsulfóxido (DMSO) a una concentración de 60 mg/L. Listeria monocytogenes ICTA-12446, fue inoculado en caldo nutriente junto con soluciones del extracto a diferentes concentraciones (20, 30, 40, 50 y 60 mg/L), se emplearon tiempos de contacto (2.5, 5, 10 y 15 minutos). Cumplido cada tiempo se realizaron diluciones seriadas e inocularon en agar nutritivo por extensión durante 24 h a 37ºC. Se efectuó el recuento en Unidades Formadoras de Colonias UFC. Al comparar las concentraciones del extracto se evidencia entre 20 y 60 mg/mL diferencia significativa, mientras que en 30, 40 y 50 mg/mL un comportamiento similar. Al contrastar tiempos de contacto, se observa que entre el tiempo 2.5 min y los restantes un p=0,03. El tiempo mínimo donde existió inhibición fue 2.5 minutos, y concentración mínima inhibitoria de 20 mg/mL. Los cuatro tiempos de contacto arrojan porcentajes de inhibición microbiana de 100% al emplear 60mg/mL. Se concluye que el extracto etanólico foliar de neem posee un efecto inhibitorio sobre Listeria monocytogenes(AU)

Listeria monocytogenes is a pathogen causing foodborne illness. In seeking to control its spread using natural substances in order to show if the leaf ethanol extract of neem (Azadirachta indica A. Juss) has antimicrobial effect on L. monocytogenes ICTA-12446, was proposed. The extract was obtained from neem leaves, which was subjected to drying for 8 days. It was reduced in size mechanically, and subjected to cold soak for 3 days, using 96% ethanol in amber vessels, filtered and concentrated in rot evaporator. Concentrated was solubilized with dimethylsulfoxide (DMSO) and standarized to achieve a concentration of 60 mg/mL Listeria monocytogenes was inoculated in nutrient broth with extract solutions at different concentrations (20, 30, 40, 50 and 60mg/mL), four contact times (2.5, 5, 10 and 15 minutes) were used. Completed each time it was diluted and inoculated on nutrient agar by extension for 24h at 37ºC. The count of Colony Forming Units UFC was taking. Comparing the concentrations of the extract between 20 and 60mg /mL significant difference was appreciate, while 30, 40 and 50 mg/mL show a similar behavior. Contrasting contact times observed between time 2.5 min and the remaining p = 0.03. The minimum time where there was some kind of inhibition was 2.5 minutes, and minima inhibitory concentration of 20mg/mL. The four contact times yield microbial inhibition percentages of 100% by using 60mg/L. It is concluded that ethanol extract of neem leaf has an inhibitory effect on L. monocytogenes(AU)

In Vitro Scolicidal Effects of Salvadora persica Root Extract against Protoscolices of Echinococcus granulosus

It has been known that Arak, Salvadora persica, has a number of medicinal properties. We tried to investigate in vitro scolicidal effect of root extracts of this plant against protoscolices from hydatid cysts of Echinococcus granulosus. Protoscolices were aseptically collected from sheep livers containing hydatid cysts. S. persica root extract was used in 10, 30, and 50 mg/ml concentration for 10, 20, and 30 min. The viability of protoscolices was ascertained by 0.1% eosin staining. Scolicidal activity of S. persica extract at a concentration of 10 mg/ml was 36.3%, 50.3%, and 70.8% after 10, 20, and 30 min of exposure, respectively. The scolicidal effect of this extract at a concentration of 30 mg/ml was 52.9%, 86.7%, and 100% after 10, 20, and 30 min of exposure, respectively. S. persica extract at a concentration of 50 mg/ml, meanwhile, killed 81.4%, 100%, and 100% of protoscolices after 10, 20, and 30 min, respectively. Also, the cytotoxic potential of S. persica was assessed on human liver cells (HepG2) using trypan blue exclusion test. No cytotoxic effect was observed on HepG2 cell line. The present study confirmed for the first time that the ethanolic extract of S. persica has high scolicidal power in vitro. However, in vivo effect of this material remains to be studied for treatment of echinococcosis in humans and herbivorous animals.

Is the ethanol wet-bonding technique a promising one? / ¿Es prometedora la técnica de adhesión húmeda en etanol?

The ethanol wet-bonding technique (EWBT) was introduced in an attempt to overcome the problems caused by high hydrophilicity and/or incomplete penetration of most commercially available adhesive systems. This strategy provides better conditions for the inter-diffusion of hydrophobic dentin monomers. Today, there are many EWBT protocols, which yield bonding interfaces with minimal degradation and longer durability compared with commercial hydrophilic adhesive systems. The aim of this review is to discuss in greater detail the EWBT, focused on the following aspects: dentin saturation, hydrophobic primer preparation, inactivation of metalloproteinases (MMPs), dentin biomimetic remineralization and the clinical perspectives of this technique. The present review on the EWBT provides support for a better understanding of the behavior of dentin when exposed to dehydration and hydrophobic monomer interaction. Moreover, additional studies are suggested to investigate the long-term stability of this type of hybrid layer.

La técnica de la adhesión húmeda en etanol (TAHE) se introdujo en un intento de superar los problemas causados por la alta hidrofilicidad y/o la penetración incompleta de la mayoría de los sistemas adhesivos disponibles comercialmente. Esta estrategia ofrece mejores condiciones para la interdifusión de monómeros dentinarios hidrofóbicos. Hoy en día, hay muchos protocolos TAHE que producen las interfaces de unión con mínima degradación y mayor durabilidad en comparación con los sistemas adhesivos hidrofilicos comerciales. El objetivo de esta revisión es discutir con más detalle la TAHE, explicando los siguientes aspectos relacionados: la saturación de la dentina, la preparación del primer hidrofóbico, la inactivación de las metaloproteinasas (MMP's), remineralización biomimética de la dentina, y las perspectivas clínicas de esta técnica. La presente revisión sobre la TAHE proporciona soporte para una mejor comprensión del comportamiento de la dentina cuando es expuesto a la deshidratación y la interacción con monómero hidrófobo. Además, se sugieren estudios adicionales para investigar la estabilidad a largo plazo de este tipo de camada híbrida.

A simple and efficient method for extraction of Taq DNA polymerase

Background Thermostable DNA polymerase (Taq Pol ?) from Thermus aquaticus has been widely used in PCR, which was usually extracted with Pluthero's method. The method used ammonium sulfate to precipitate the enzyme, and it saved effort and money but not time. Moreover, we found that 30-40% activity of Taq Pol I was lost at the ammonium sulfate precipitation step, and the product contained a small amount of DNA. Results We provided a novel, simplified and low-cost method to purify the Taq Pol ? after overproduction of the enzyme in Escherichia coli, which used ethanol instead of ammonium sulfate to precipitate the enzyme. The precipitate can be directly dissolved in the storage buffer without dialysis. In addition, DNA and RNA contamination was removed with DNase I and RNase A before precipitation, and the extraction procedure was optimized. Our improvements increase recovery rate and specific activity of the enzyme, and save labor, time, and cost. Conclusions Our method uses ethanol, DNase I, and RNase A to purify the Taq Pol ?, and simplifies the operation, and increases the enzyme recovery rate and quality.

Effect of ethanolic extract of Coriandrum sativum L. on tacrine induced orofacial dyskinesia.

The effect of ethanolic extract of Coriandrum sativum L. seeds (100, 200 mg/kg) was studied on tacrine induced orofacial dyskinesia. Tacrine (2.5 mg/kg, i.p.) treated animals were observed for vacuous chewing movements (VCM), tongue protrusions (TP) and orofacial bursts (OB) for 1 h followed by observations for locomotor changes and cognitive dysfunction. Sub-chronic administration of Coriandrum sativum L. seed extract (E-CS) (100, 200 mg/kg, p.o., for 15 days significantly (P <0.05) decreased the tacrine induced VCM, TP and OB; and also significantly (P <0.05), increased locomotion and cognition compared to the tacrine treated group. Biochemical analysis revealed that tacrine administration significantly (P <0.05) decreased the levels of superoxide dismutase (SOD), Catalase (CAT), glutathione reductase (GSH) levels and also significantly (P <0.05) increased lipid peroxidation (LPO) as an index of oxidative stress, whereas sub-chronic administration of E-CS significantly (P <0.05) improved the antioxidant enzyme (i.e. SOD, CAT, and GSH) levels and also significantly (P <0.05) decreased lipid peroxidation (LPO). The results have demonstrated the protective role of ethanolic extract of Coriandrum sativum. L against tacrine induced orofacial dyskinesia.