Lipid nanoparticle delivery of siRNA targeting Cyp2e1 gene attenuates subacute alcoholic liver injury in mice / 浙江大学学报·医学版

OBJECTIVES@#To investigate the effect and mechanism of lipid nanoparticle (LNP) delivery of small interfering RNA (siRNA) targeting Cyp2e1 gene on subacute alcoholic liver injury in mice.@*METHODS@#siRNA targeting Cyp2e1 gene was encapsulated in LNP (si-Cyp2e1 LNP) by microfluidic technique and the resulting LNPs were characterized. The optimal dose of si-Cyp2e1 LNP administration was screened. Forty female C57BL/6N mice were randomly divided into blank control group, model control group, si-Cyp2e1 LNP group, LNP control group and metadoxine group. The subacute alcoholic liver injury mouse model was induced by ethanol feeding for 10 d plus ethanol gavage for the last 3 d. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and the superoxide dismutase (SOD) activity as well as malondialdehyde, reactive oxygen species, glutathione, triacylglycerol, total cholesterol contents in liver tissue were measured in each group, and liver index was calculated. The expression of genes related to oxidative stress, lipid synthesis and inflammation in each group of mice were measured by realtime RT-PCR.@*RESULTS@#Compared with the model control group, the levels of liver index, serum ALT, AST activities, malondialdehyde, reactive oxygen species, triacylglycerol, total cholesterol contents in liver tissue decreased, but the SOD activity as well as glutathione increased in the si-Cyp2e1 LNP group (all P<0.01). Hematoxylin-eosin staining result showed disorganized hepatocytes with sparse cytoplasm and a large number of fat vacuoles and necrosis in the model control group, while the si-Cyp2e1 LNP group had uniformly sized and arranged hepatocytes with normal liver tissue morphology and structure. Oil red O staining result showed si-Cyp2e1 LNP group had lower fat content of the liver compared to the model control group (P<0.01), and no fat droplets accumulated. Anti-F4/80 monoclonal antibody fluorescence immunohistochemistry showed that the si-Cyp2e1 LNP group had lower cumulative optical density values compared to the model control group (P<0.01) and no significant inflammatory reaction. Compared with the model control group, the expression of catalytic genes P47phox, P67phox and Gp91phox were reduced (all P<0.01), while the expression of the antioxidant enzyme genes Sod1, Gsh-rd and Gsh-px were increased (all P<0.01). The mRNA expression of the lipid metabolism genes Pgc-1α and Cpt1 were increased (all P<0.01) and the lipid synthesis-related genes Srebp1c, Acc and Fasn were decreased (all P<0.01); the expression of liver inflammation-related genes Tgf-β, Tnf-α and Il-6 were decreased (all P<0.01).@*CONCLUSIONS@#The si-Cyp2e1 LNP may attenuate subacute alcoholic liver injury in mice mainly by reducing reactive oxygen levels, increasing antioxidant activity, blocking oxidative stress pathways and reducing ethanol-induced steatosis and inflammation.

Study on the effect of small alcohol on lipid hydration and liposome formation / 生物医学工程学杂志

Liposomes with precisely controlled composition are usually used as membrane model systems to investigate the fundamental interactions of membrane components under well-defined conditions. Hydration method is the most common method for liposome formation which is found to be influenced by composition of the medium. In this paper, the effects of small alcohol (ethanol) on the hydration of lipid molecules and the formation of liposomes were investigated, as well as its coexistence with sodium chloride. It was found that ethanol showed the opposite effect to that of sodium chloride on the hydration of lipid molecules and the formation of liposomes. The presence of ethanol promoted the formation of liposomes within a certain range of ethanol content, but that of sodium chloride suppressed the liposome formation. By investigating the fluorescence intensity and continuity of the swelled membranes as a function of contents of ethanol and sodium chloride, it was found that sodium chloride and ethanol showed the additive effect on the hydration of lipid molecules when they coexisted in the medium. The results may provide some reference for the efficient preparation of liposomes.

Intrauterine alcohol exposure disturbs trabecular morphology in the Sprague Dawley rat humerus epiphysis up to 3- weeks postnatally / La exposición al alcohol intrauterino altera la morfología trabecular en la epífisis del húmero de la rata Sprague Dawley hasta 3 semanas después del nacimiento

SUMMARY: Gestational alcohol exposure inhibits neurological as well as bone growth and development both in fetal and postnatal life. Stunted stature, osteoporosis and fractures in adult life are some of the adverse effects. While the impact of intrauterine alcohol on the brain has been extensively investigated, studies on the effects on bone are relatively few. Therefore, our study aimed to examine the impact of prenatal alcohol exposure on bone microarchitecture in 3-week-old rats using Micro-focus X-Ray Computed Tomography (Micro CT). Time mated pregnant Sprague Dawley dams (13) were randomly placed into 3 groups: ethanol (n=5), saline control (n=5) and untreated control (n=3). The former 2 groups received treatment with 0.015ml/g of 25.2 % ethanol and 0.9 % saline, respectively, for the first 19 days of gestation. The untreated group received no treatment. The pups remained with their dams until termination at 21 days of age. From each dam, 2 pups were collected resulting in: ethanol (n=10), saline controls (n= 10) and untreated controls (n = 6). The humeri of the pups were dissected and scanned using a 3D-μCT scanner (Nikon XTH 225L) at 15μm resolution. Trabecular and cortical parameters were analysed using Volume Graphics Studio® software following reconstruction. Results showed a decrease in trabecular size, spaces, thickness, and volume. There was a decrease in cortical bone area in the ethanol group compared to the controls. These findings may suggest that osteoporosis and fractures seen as gestational alcohol effects may be due to compromised trabecular structure.

RESUMEN: La exposición al alcohol durante la gestación inhibe el crecimiento y desarrollo neurológico y óseo tanto en la vida fetal como posnatal. Algunos de los efectos adversos incluyen la estatura atrofiada, osteoporosis y fracturas en la vida adulta. Si bien se ha estudiado el impacto del alcohol intrauterino en el cerebro, los estudios sobre los efectos en los huesos son escasos. Por lo tanto, nuestro estudio tuvo como objetivo examinar el impacto de la exposición prenatal al alcohol en la microarquitectura ósea en ratas de 3 semanas de edad utilizando Tomografía Computarizada de Rayos X Micro-focus (Micro CT). Las hembras de Sprague Dawley preñadas con apareamiento temporal (13) se colocaron aleatoriamente en 3 grupos: etanol (n = 5), control de solución salina (n = 5) y control sin tratar (n = 3). Los primeros 2 grupos recibieron tratamiento con 0,015 ml /g de etanol al 25,2 % y solución salina al 0,9 %, respectivamente, durante los primeros 19 días de gestación. El grupo no tratado no recibió tratamiento. Las crías permanecieron con sus madres hasta la terminación a los 21 días de edad. De cada madre, se recolectaron 2 crías que dieron como resultado: etanol (n = 10), controles salinos (n = 10) y controles no tratados (n = 6). Se diseccionaron y escanearon los húmero de las crías usando un escáner 3D-μCT (Nikon XTH 225L) a una resolución de 15 μm. Los parámetros trabeculares y corticales se analizaron utilizando el software Volume Graphics Studio® después de la reconstrucción. Los resultados mostraron una disminución en el tamaño trabecular, los espacios, el grosor y el volumen. Hubo una disminución en el área del hueso cortical en el grupo de etanol en comparación con los controles. Estos hallazgos pueden sugerir que la osteoporosis y las fracturas por causa de los efectos del alcohol gestacional se pueden deber a una estructura trabecular comprometida.

Actividad antibacteriana de un extracto etanólico de propóleo peruano frente a Streptococcus mutans / Antibacterial activity of a Peruvian propolis ethanolic extract against Streptococcus mutans

Introducción: Los derivados del propóleo poseen propiedades antimicrobianas importantes y presentan un potencial uso para la prevención y tratamiento de la caries dental. Objetivo: Evaluar la actividad antibacteriana de un extracto etanólico de propóleo peruano frente a Streptococcus mutans. Métodos: Se obtuvo el extracto etanólico de propóleo por maceración en alcohol al 70 por ciento durante 15 días. El extracto etanólico de propóleo fue diluido con agua destilada para obtener concentraciones de 75 por ciento, 50 por ciento y 25 por ciento. La actividad antibacteriana se evaluó mediante la prueba de difusión en disco sobre medio agar cerebro-corazón inoculado con S. mutans ATCC® 25175™, se empleó clorhexidina (CHX) al 0,12 por ciento como control. Las placas de Petri fueron incubadas por 48 horas a 37 ºC en condiciones de microaerofilia. Posteriormente se realizó la medición de los halos de inhibición con un compás Vernier. Resultados: Todas las concentraciones del extracto etanólico de propóleo presentaron actividad antibacteriana frente al S. mutans (25 por ciento = 17,582 ± 2,578 mm; 50 por ciento = 16,906 ± 1,892 mm; 75 por ciento = 16,881 ± 2,013 mm; 100 por ciento = 17,201 ± 1,305 mm). Sin embargo, fueron menores que la CHX al 0,12 por ciento (24,543 ± 2,486 mm) (p < 0,05). Según la escala de Duraffourd, S. mutans fue sensible (+) y muy sensible (++) para todas las concentraciones del extracto etanólico de propóleo, mientras que para CHX al 0,12 % fue sumamente sensible (+++) (p < 0,05). Conclusiones: El extracto etanólico de propóleo peruano presenta actividad antibacteriana significativa considerada como sensible y muy sensible frente a S. mutans(AU)

Introduction: Due to their important antimicrobial properties, propolis by-products are potentially useful for the prevention and treatment of dental caries. Objective: Evaluate the antibacterial activity of a Peruvian propolis ethanolic extract against Streptococcus mutans. Methods: The propolis ethanolic extract was obtained by maceration in 70 percent alcohol for 15 days. The extract was diluted in distilled water to obtain concentrations of 75 percent , 50 percent and 25 percent . Antibacterial activity was evaluated by the disk diffusion test in brain heart agar medium inoculated with S. mutans ATCC® 25175™. Chlorhexidine (CHX) 0.12 percent was used as control. The Petri plates were incubated for 48 hours at 37ºC in microaerophilic conditions. The inhibition haloes were then measured with a Vernier caliper. Results: All the concentrations of the propolis ethanolic extract displayed antibacterial activity against S. mutans: 25 percent = 17.582 ± 2.578 mm; 50 percent = 16.906 ± 1.892 mm; 75 percent = 16.881 ± 2.013 mm; 100 percent = 17.201 ± 1.305 mm. However, values were lower than those of 0.12 percent CHX: 24.543 ± 2.486 mm (p < 0.05). According to the Duraffourd scale, S. mutans was sensitive (+) and very sensitive (++) to all propolis ethanolic extract concentrations, and highly sensitive to 0.12 percent chlorhexidine (+++) (p < 0.05). Conclusions: The Peruvian propolis ethanolic extract displays significant antibacterial activity against S. mutans. Such activity was evaluated as sensitive and very sensitive(AU)

Alcohol (70%) versus alcoholic chlorhexidine solution (0. 5%) in skin antisepsis for neuraxial blocks: a randomized clinical trial / Álcool 70% versus solução alcoólica de clorexidina 0, 5% na antissepsia da pele para bloqueios do neuroeixo: ensaio clínico randomizado

ABSTRACT Objective: to compare the use of 0.5% alcoholic chlorhexidine and 70% alcohol in skin antisepsis for neuraxial blocks. Method: this is a non-inferiority randomized clinical trial, with two parallel arms. Seventy patients who were candidates for neuraxial block were randomly allocated to group A (n = 35), in whom antisepsis was performed with 0.5% alcoholic chlorhexidine, or to group B (n = 35), in whom we used 70% hydrated ethyl alcohol. Swabs were harvested for culture at three times: before antisepsis, two minutes after application of the antiseptic, and immediately after puncture. The samples were sown in three culture media and the number of colony forming units (CFU) per cm² was counted. Results: there was no difference between the groups regarding age, sex, body mass index, time to perform the block or type of block. There were no differences between groups in the CFU/cm² counts before antisepsis. There was less bacterial growth in group B two minutes after application of the antiseptic (p = 0.048), but there was no difference between the groups regarding the number of CFU/cm² at the end of the puncture. Conclusion: 70% alcohol was more effective in reducing the number of CFU/cm² after two minutes, and there was no difference between the two groups regarding skin colonization at the end of the procedure. These results suggest that 70% alcohol may be an option for skin antisepsis before neuraxial blocks. Trial registration: ClinicalTrials.gov, NCT02833376.

RESUMO Objetivo: comparar o uso de solução alcoólica de clorexidina 0,5% e de álcool 70% na antissepsia da pele para bloqueios do neuroeixo. Método: ensaio clínico randomizado de não inferioridade, com dois braços paralelos. Foram selecionados 70 pacientes candidatos à bloqueio do neuroeixo, randomicamente alocados para o grupo A (n=35), em que a antissepsia foi realizada com clorexidina alcoólica 0,5%, ou para o grupo B (n=35), em que se utilizou álcool etílico hidratado 70%. Foram coletadas, com swab, amostras para cultura em três momentos: antes da antissepsia, dois minutos após aplicação do antisséptico, e imediatamente após a punção. As amostras foram semeadas em três meios de cultura e foi contabilizado o número de unidades formadoras de colônias (UFC) por cm². Resultados: não houve diferença entre os grupos quanto à idade, ao sexo, ao índice de massa corporal, ao tempo para realização do bloqueio ou tipo de bloqueio. Também não houve diferenças entre os grupos na contagem de UFC/cm² antes da antissepsia. Constatou-se menor crescimento bacteriano no grupo B dois minutos após aplicação do antisséptico (p=0,048), mas não houve diferença entre os grupos quanto ao número de UFC/cm² ao final da punção. Conclusão: o álcool 70% mostrou-se mais efetivo em reduzir o número de UFC/cm² após dois minutos, e não houve diferença entre os dois grupos quanto à colonização da pele ao final do procedimento. Esses resultados sugerem que o álcool 70% pode ser opção para antissepsia da pele antes de bloqueios do neuroeixo. Registro ensaio clínico: ClinicalTrials.gov, NCT02833376.

Brain structure morphology after being fixated with ethanol on electron microscope / Morfología de la estructura cerebral después de la fijación con etanol en microscopio electrónico

Fixation is one of the processes in preparing histology and pathology. The common material for fixation is buffered formalin including paraformaldehyde. However, the effect of the damaged cells, which is fixed for a long time, causes the research for other fixation materials to become necessary. In addition, paraformaldehyde is also harmful to human body and natural environment. Ethanol is one of the alternative fixation materials, which has been used for two hundred years. It has been used for many purposes, both in routine staining and immunohistochemistry. Nonetheless, no research confirms its effect on the electron microscope. The authors studied the effect of 50 % of ethanol on the cell membrane, organelles, and nucleus of Purkinje cells (Neuron purkinjense) observed on a light microscope and Transmitted Electron Microscope (TEM). Then it was compared to buffered formalin. In the light microscope, it shows that both of fixations have no different effects of the morphology of the cell membrane, cytoplasm, the nucleus of Purkinje cells and the neutrophils. We assume that our 50 % of ethanol concentration is almost the same as BF 10 % in the ability of hardening tissue and color absorption based on the previous study. In TEM, the structure of the cell membrane, organelles, and cytoplasm of Purkinje cell look broken in the cerebellum of 50 % of ethanol except for the nucleus. There was no significant difference diameter of the nucleus. It happened in general because of the shrinkage effect of ethanol. However, the authors recommend using 50 % of ethanol for routine staining.

La fijación es uno de los procesos en la preparación de muestras para histología y patología. El material más común para la fijación es la formalina tamponada. Sin embargo, el daño a las células que se mantienen en formalina durante mucho tiempo, hace necesario buscar otros materiales de fijación. Además, el paraformaldehido también es perjudicial para el cuerpo humano y el medio ambiente natural. El etanol es uno de los materiales de fijación alternativos que se ha utilizado durante muchos años, con diversos objetivos, tanto en la tinción de rutina como en la inmunohistoquímica. Sin embargo no se ha confirmdo su efecto con microscopio electrónico. Los autores estudiaron el efecto del 50 % de etanol sobre la membrana celular, los orgánulos y el núcleo de las células de Purkinje observados en un microscopio óptico y un microscopio de transmisión electrónico (TEM). Luego se comparó con la formalina tamponada. En el microscopio óptico se observó que ambas fijaciones no tienen efectos diferentes a la morfología de la membrana celular, el citoplasma, el núcleo de las células de Purkinje y los neutrófilos. Suponemos que nuestra concentración de 50 % de etanol es casi la misma que BF 10 % en la capacidad de endurecer el tejido y la absorción de color según el estudio anterior. En TEM, la estructura de la membrana celular, los orgánulos y el citoplasma de la célula de Purkinje presentaban daño en el cerebelo con un 50 % de etanol, a excepción del núcleo. No hubo diferencia significativa en el diámetro del núcleo. En general lo anterior se debió al efecto de contracción del etanol. En conclusión los autores recomiendan usar 50% de etanol para la tinción de rutina.

Effect of ethanol extracts of Antrodia cinnamomea on head and neck squamous cell carcinoma cell line

Head and neck squamous cell carcinoma (HNSCC) is one of the most common malignant tumors. Ethanol extract of Antrodia cinnamomea (EEA) has been widely studied for its health benefits including anticancer effects. The purpose of this study was to assess the effects of EEA on HNSCC. Cell proliferation, transwell, and wound healing assays were performed. The impact of EEA on tumor growth was investigated using a xenograft model. Expressions of migration-related proteins (MMP-2, MMP-9, TIMP-1, and TIMP-2) and apoptosis-related proteins (cleaved caspase-9 and cleaved PARP) were determined using western blot analysis. The results indicated that EEA significantly inhibited the capacities of proliferation, invasion, and migration of HNSCC cells in a dose-dependent manner. Cleaved caspase-9 and cleaved PARP expressions were increased in cells treated with an increasing concentration of EEA, which suggested that EEA induced apoptosis of HNSCC. MMP-2 and MMP-9 were downregulated when cells were administered EEA, while TIMP-1 and TIMP-2 were not affected, which uncovered the mechanisms mediating the EEA-induced inhibition on cell invasion and migration. The animal experiment also suggested that EEA inhibited tumor growth. Our study confirmed the inhibitive effects of EEA on cell proliferation, invasion, and migration of HNSCC in vitro and in vivo, providing the basis for further study of the application of EEA as an effective candidate for cancer treatment.

Effect of ethanol-wet bonding on porosity and retention of fiberglass post to root dentin

Abstract: This in vitro study aimed to assess the effects of different pretreatments used to adapt glass-fiber posts cemented to root canals with different resin cements, regarding porosity percentage and bond strength. Twelve bovine incisor roots were prepared with Largo drills. After post space preparation, the specimens were randomized into two types of pretreatment groups (n = 3): water-wet bonding and ethanol-wet bonding. After the post were cemented, the roots were stored in 100% humidity at room temperature for 7 days. The samples were scanned by microcomputed tomography (micro-CT). Images were reconstructed by NRecon software, and CTAn software was used to analyze the porosity percentage (%) at the luting interface. Evaluation of the push-out bond strength was performed by serially cutting the roots, and submitting the slices to testing. Additionally, the resin cement post-gel shrinkage values (%) were measured using the strain-gauge method (n = 10). Data were analyzed by two-way ANOVA, Tukey's test and Student's t test (a = 0.05). The roots prepared with ethanol-wet bonding using RelyX™ U200 had significantly lower porosity in the coronal and apical thirds (p < 0.05). The group prepared with ethanol-wet bonding using RelyX™ ARC presented better bond strength results in the coronal and apical thirds (p < 0.05). RelyX™ ARC (0.97%) produced a higher post-gel shrinkage value than RelyX™ U200 (0.77%). Canals pretreated with ethanol-wet bonding presented better outcomes in regard to porosity percentage and push-out bond strength.

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.

Reduction of the DNA damages, Hepatoprotective Effect and Antioxidant Potential of the Coconut Water, ascorbic and Caffeic Acids in Oxidative Stress Mediated by Ethanol

ABSTRACT Hepatic disorders such as steatosis and alcoholic steatohepatitis are common diseases that affect thousands of people around the globe. This study aims to identify the main phenol compounds using a new HPLC-ESI+-MS/MS method, to evaluate some oxidative stress parameters and the hepatoprotective action of green dwarf coconut water, caffeic and ascorbic acids on the liver and serum of rats treated with ethanol. The results showed five polyphenols in the lyophilized coconut water spiked with standards: chlorogenic acid (0.18 µM), caffeic acid (1.1 µM), methyl caffeate (0.03 µM), quercetin (0.08 µM) and ferulic acid (0.02 µM) isomers. In the animals, the activity of the serum γ-glutamyltranspeptidase (γ-GT) was reduced to 1.8 I.U/L in the coconut water group, 3.6 I.U/L in the ascorbic acid group and 2.9 I.U/L in the caffeic acid groups, when compared with the ethanol group (5.1 I.U/L, p<0.05). Still in liver, the DNA analysis demonstrated a decrease of oxidized bases compared to ethanol group of 36.2% and 48.0% for pretreated and post treated coconut water group respectively, 42.5% for the caffeic acid group, and 34.5% for the ascorbic acid group. The ascorbic acid was efficient in inhibiting the thiobarbituric acid reactive substances (TBARS) in the liver by 16.5% in comparison with the ethanol group. These data indicate that the green dwarf coconut water, caffeic and ascorbic acids have antioxidant, hepatoprotective and reduced DNA damage properties, thus decreasing the oxidative stress induced by ethanol metabolism.

Draft genome sequence of alcohol-tolerant bacteria Pediococcus acidilactici strain K3

Abstract Pediococcus acidilactici strain K3 is an alcohol-tolerant lactic acid bacterium isolated from nuruk, which is a traditional Korean fermentation starter for makgeolli brewing. Draft genome of this strain was approximately 1,991,399 bp (G+C content, 42.1%) with 1525 protein-coding sequences (CDS), of which 44% were assigned to recognized functional genes. This draft genome sequence data of the strain K3 will provide insights into the genetic basis of its alcohol-tolerance.

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.

Calcium hydroxide associated with a new vehicle: Psidium cattleianum leaf extracts. Tissue response evaluation

Abstract The aim of this study was to evaluate edemogenic activity and subcutaneous inflammatory reaction induced by Psidium cattleianum leaf extracts associated with Ca(OH)2. Thirty male Wistar rats, split equally into three groups [aqueous extract + Ca(OH)2; ethanolic extract + Ca(OH)2; and propylene glycol + Ca(OH)2], were assessed every 3 h or 6 h (five animals in each period). Under general anesthesia, 0.2 mL of 1% Evans blue per 100 g of body weight was injected into the penile vein and each combination to be evaluated was subcutaneously injected into the dorsal region 30 min thereafter. Edemogenic activity was analyzed by spectrophotometry (λ=630 nm). For inflammatory reaction analysis, 50 rats received four polyethylene tubes (three experimental groups) and an empty tube (control group). The assessments were made at 7, 15, 30, 60, and 90 days, followed by hematoxylin-eosin staining and by the assignment of scores for evaluation of tissue response intensity. Ethanolic extract + Ca(OH)2 yielded the largest edemogenic activity at 3 h. Intergroup differences at 6 h were not significant. The histological analysis showed progressive repair over time (p<0.05) and aqueous and ethanolic extracts produced similar responses to those of the control and Ca(OH)2 + propylene glycol groups. Psidium cattleianum leaf extracts used as Ca(OH)2 vehicles evoked similar tissue response when compared to Ca(OH)2 associated with propylene glycol.

Morphological and immunohistochemical analysis of apoptosis in the cerebellum of rats subjected to focal cerebral ischemia with or without alcoholism model

ABSTRACT PURPOSE: To evaluated histopathological changes, morphometric and expression of proteins CASPASE-3, BCL-2 and XIAP related to apoptosis in the cerebellum after induction of temporary focal cerebral ischemia followed by reperfusion, with or without a model of chronic alcoholism. METHODS: Fifty Wistar rats were used and divided into: control group (C), sham group (S), ischemic group (I), alcoholic group (A), and ischemic and alcoholic group (IA). The cerebellum samples collected were stained for histopathological and morphometric analysis and immunohistochemistry study. RESULTS: Histopathological changes were observed a greater degree in animals in groups A and IA. The morphometric study showed no difference in the amount of cells in the granular layer of the cerebellum between the groups. The expression of CASPASE-3 was higher than BCL-2 and XIAP in the groups A and IA. CONCLUSION: We observed correlation between histopathological changes and the occurrence of apoptosis in cerebellar cortex.

Anti-inflammatory effects of Salvia plebeia R. Br extract in vitro and in ovalbumin-induced mouse model

BACKGROUND: Asthma is an increasing global health problem, and novel strategies to prevent or ameliorate the condition are needed. Here, the effects of 80 % ethanol extracts of Salvia plebeia R. Br. (SE) on an induced inflammatory response were investigated RESULTS: Salvia plebeia R. Br. inhibited production of pro-inflammatory cytokines, such as TNF-α and IL-6, as well as nitric oxide (NO) in LPS-stimulated RAW 264.7 cells. NO and pro-inflammatory cytokine production was suppressed more effectively by SE of the aerial parts (SE-A) than of the roots (SE-R) of S. plebeia. In BEAS-2B cells, both SE-A and SE-R inhibited the increase in production of the inflammatory cytokines IL-6 and IL-8. We also investigated the antiasthmatic effects of SE in an ovalbumin (OVA)-induced BALB/c mouse model. SE-A treatment significantly reduced the number of airway eosinophils, IL-4 and IL-13 levels, mucus production, and inflammatory infiltration, as compared with the corresponding levels in the untreated, OVA-induced mice, and had similar effects to dexamethasone CONCLUSIONS: Salvia plebeia ethanol extract ameliorated the induced inflammatory response in RAW 264.7 and BEAS-2B cells, with more effective inhibition noted for SE-A than for SE-R. SE-A treatment was effective in improving the histopathological changes in the lungs of asthma model mice via modulation of eosinophils and Th2 cytokines. These results suggest that SE-A can be considered as a therapeutic agent that can potentially relieve asthma

Effects of ethanol consumption and alcohol detoxification on the biomechanics and morphology the bone in rat femurs / Efeitos do consumo de etanol e da desintoxicação alcóolica sobre a biomecânica e morfologia óssea em fêmur de ratos

Abstract The objective of this study was to verify the effects of ethanol consumption and alcohol detoxification on the biomechanics, area and thickness of cortical and trabecular bone in rat femur. This was an experimental study in which 18 male Wistar rats were used, with 40 days of age, weighing 179±2.5 g. The rats were divided into three groups (n=06): CT (control), AC (chronic alcoholic), DT (detoxification). After experimental procedures, the animals were euthanized by an overdose of the anesthetic and their femurs were collected for mechanical testing and histological processing. All animals did not present malnutrition or dehydration during experimentation period. Morphometric analysis of cortical and trabecular bones in rat femurs demonstrated that AC animals showed inferior dimensions and alcohol detoxification (DT) allowed an enhancement in area and thickness of cortical and trabecular bone. Material and structural properties data of AC group highlighted the harmful effects of ethanol on bone mechanical properties. The results of this study demonstrated that chronic alcoholic rats (AC) presented major bone damage in all analyzed variables. Those findings suggested that alcohol detoxification is highly suggested in pre-operative planning and this corroborates to the success of bone surgery and bone tissue repair. Thanks to the financial support offered by PROBIC – UNIFENAS.

Resumo O objetivo deste estudo foi verificar os efeitos do consumo de etanol e da desintoxicação alcoólica sobre a biomecânica, área e espessura do osso cortical e trabecular em fêmur de ratos. Este foi um estudo experimental no qual foram utilizados 18 ratos Wistar machos, com 40 dias de vida, pesando 179±2,5 g. Os ratos foram divididos em três grupos (n=06): CT (controle), AC (alcoolista crônico), e DT (desintoxicado). Após os procedimentos experimentais os animais foram eutanaziados por uma overdose de anestésico e os fêmures coletados para os testes mecânicos e processamento histológico. Todos os animais não apresentaram desnutrição ou desidratação durante o período de experimentação. As análises morfométrica do osso cortical e trabecular demonstraram que os animais do grupo AC apresentavam dimensões inferiores, enquanto nos animais do grupo DT observou-se um aumento na área e espessura do osso cortical e trabecular. Dados dos materiais e das propriedades estruturais óssea do grupo AC destacam os efeitos nocivos do etanol sobre as propriedades mecânicas do osso. Os resultados deste estudo demonstraram que os ratos do grupo AC apresentaram danos significativos no osso em todas as variáveis ​​analisadas. Esses resultados sugerem que a desintoxicação alcoólica é recomendada no planejamento pré-operatório e isso corrobora para o sucesso de cirurgias e reparação no tecido ósseo. Agradecemos ao apoio financeiro oferecido pelo PROBIC – UNIFENAS.

A review on alcohol: from the central action mechanism to chemical dependency / Uma revisão sobre o álcool: do mecanismo de ação central à dependência química

SummaryIntroduction:alcohol is a psychotropic depressant of the central nervous system (CNS) that promotes simultaneous changes in several neuronal pathways, exerting a profound neurological impact that leads to various behavioral and biological alterations.Objectives:to describe the effects of alcohol on the CNS, identifying the signaling pathways that are modified and the biological effects resulting from its consumption.Methods:a literature review was conducted and articles published in different languages over the last 15 years were retrieved.Results:the studies reviewed describe the direct effect of alcohol on several neurotransmitter receptors (gamma-aminobutyric acid [GABA], glutamate, endocannabinoids AEA and 2-AG, among others), the indirect effect of alcohol on the limbic and opioid systems, and the effect on calcium and potassium channels and on proteins regulated by GABA in the hippocampus.Discussion and conclusion:the multiple actions of alcohol on the CNS result in a general effect of psychomotor depression, difficulties in information storage and logical reasoning and motor incoordination, in addition to stimulating the reward system, a fact that may explain the development of addiction. Knowledge on the neuronal signaling pathways that are altered by alcohol allows the identification of effectors which could reduce its central action, thus, offering new therapeutic perspectives for the rehabilitation of alcohol addicts.

ResumoIntrodução:o álcool é uma substância psicotrópica depressora do sistema nervoso central (SNC), que promove alteração simultânea de inúmeras vias neuronais, gerando profundo impacto neurológico e traduzindo-se em diversas alterações biológicas e comportamentais.Objetivos:descrever as ações do álcool sobre o SNC, identificando as vias de sinalização modificadas e os efeitos biológicos gerados pelo seu consumo.Métodos:revisão bibliográfica, priorizando trabalhos multilinguísticos publicados nos últimos 15 anos.Resultados:são descritas ação direta do álcool em inúmeros receptores de neurotransmissores (ácido gama-aminobutírico – GABA, glutamato, endocanabinoides AEA e 2-AG, entre outros), ação indireta do álcool no sistema límbico e opioide, ação sobre canais de cálcio, potássio e proteínas reguladas por GABA no hipocampo, além de ações centrais mediadas pela deficiência de vitamina B1.Conclusão:a ação multifocal do álcool sobre o SNC resulta em efeito geral de depressão psicomotora, dificuldades no armazenamento de informações e no raciocínio lógico, incoordenação motora, além da estimulação do sistema de recompensa, o que pode explicar o desenvolvimento da dependência química. O conhecimento das vias de sinalização neuronais alteradas pelo álcool permite reconhecer a descrição de efetores que possam reduzir sua ação central e, assim, vislumbrar novas perspectivas terapêuticas para a reabilitação de adictos a essa substância.

Modulation of the erythromycin resistance in Staphylococcus aureus by ethanolic extracts of Ximenia americana L and Schinopsis brasiliensis Engl / Modulación de la resistencia a la eritromicina en Staphylococcus aureus por extractos etanólicos de Ximeniaamericana L y Schinopsis brasiliensis Engl

It was evaluated the in vitro efficacy of ethanolic extract of leaves and bark of Ximenia americana L and Schinopsis brasiliensis Engl. alone and in association with erythromycin as modulators of microbial resistance against six clinical isolates of Staphylococcus aureus resistant to erythromycin (SA1-SA6) and S. aureus ATCC 25923 by the microdilution method. The extracts were also subjected to bioassay with Artemia salina. The ethanolic extract of barks of X. americana showed a synergistic effect with erythromycin against SA01, SA03 and SA04. The leaf extract of S. brasiliensis exerted synergistic effect against SA03 and the bark extract showed against SA01 and S03. The results suggest that extracts from S.brasiliensis and X. americana have potential as modulator agents of bacterial resistance, which could be used as adjuvants in the treatment of infections by S. aureus resistant to erythromycin, with previous studies of toxicity.

Se evaluó la eficacia in vitro de los extractos etanólicos de hojas y corteza de Ximenia americana L y Schinopsis brasiliensis Engl solos y en asociación con eritromicina como moduladores de la resistencia microbiana frente a seis aislados clínicos de Staphylococcus aureus resistentes a Eritromicina (SA1-SA6) y S. aureus ATCC 25923, por el método de microdilución. Además se determinó la actividad tóxica de los extractos contra Artemia salina. Solo el extracto etanólico de la corteza de X. americana mostró un efecto sinérgico con la eritromicina frente a SA01, SA03 y SA04. El extracto de las hojas de S. brasiliensis ejerció efecto sinérgico contra SA03 y el extracto de corteza, contra SA01 y S03. Los resultados sugieren que S. brasiliensis y X. americana tienen potencial como agentes moduladores de la resistencia bacteriana, que podrían ser utilizados como adyuvantes en el tratamiento de infecciones por S. aureus resistentes a eritromicina, con estudios previos de toxicidad.

Repeated binge-like ethanol administration during adolescence cause decreased C-Fos immunoreactivity in amygdala and arcuate nucleus in adult Sprague-Dawley rats / La administración repetida de etanol, tipo atracón, durante la adolescencia provoca descenso en la expresión de C-Fos en la amígdala y núcleo arqueado de ratas Sprague-Dawley adultas

Binge alcohol drinking during adolescence has been associated with neurotoxicity and increased risk for the development of alcohol use disorders. There is evidence that acute and chronic ethanol administration alters c-fos expression, an indirect index of cellular activity, in different brain regions in adult rats. We evaluate here if a binge-like pattern of ethanol exposure during adolescence has a relevant impact on basal and/or ethanol-stimulated regional c-fos activity during adulthood. For that aim, Sprague-Dawley rats PND 25 were saline pre-treated, (SP group) or binge-ethanol pre-treated (BEP group) for two­consecutive days, at 48-h intervals, over a 14-day period (PND 25 to PND 38). At adult stage (PND 63) and following 25 ethanol-free days, we evaluated c-fos immunoreactivity in response to saline or acute ethanol (1.5 or 3.0 g/kg) in the hypothalamus and amygdala. We found that acute ethanol administration dose-dependently increased c-fos activity in the the Paraventricular nucleus of the hypothalamus (PVN). Interestingly, binge-ethanol exposure during adolescence significantly reduced basal c-fos activity during adulthood in the Central nucleus of the amygdala (CeA) and the Arcuate nucleus of hypothalamus (Arc). We conclude that binge-like ethanol administration during adolescence causes long-term disturbances in basal neural activity in brain areas critically involved with ethanol consumption.

El consumo en atracón durante la adolescencia está asociado con neurotoxicidad y con el riesgo de desarrollar un trastorno en el uso de alcohol. Diversos estudios muestran que la administración aguda y crónica de alcohol en ratas adultas altera la expresión de c-fos, un marcador indirecto de actividad celular, en diferentes áreas cerebrales. Nosotros evaluamos si el patrón de consumo de alcohol en atracón durante la adolescencia tiene un impacto en la actividad basal de c-fos en esas regiones activadas por el alcohol. Utilizamos ratas Sprague-Dawley en su día post-natal 25 (PND25) tratadas con suero salino (grupo SP) o con etanol tipo atracón (grupo BEP) durante dos días consecutivos, en intervalos de 48 h, durante 14 días (PND25- PND38). En la edad adulta (PND63) y después de 25 días sin etanol, evaluamos la inmunorreactividad para c-fos en respuesta a una administración aguda de suero salino o etanol (1,5 ó 3,0 g/kg) en diferentes regiones cerebrales. La administración de alcohol incrementó de manera dosis-dependiente la actividad de c-fos en el núcleo paraventricular del hipotálamo. Además la exposición a etanol tipo atracón durante la adolescencia disminuyó la actividad basal de c-fos en la adultez en el núcleo central de la amígdala y en el núcleo arqueado del hipotálamo. Concluimos que el consumo de alcohol en atracón durante la adolescencia causa problemas a largo plazo en la actividad basal de regiones cerebrales implicadas en el consumo de alcohol.

Homeopathic drugs complementary, antidotal and inimical to Nux vomica produce stronger anti-alcoholic effect on toads than Nux

In homeopathy some drugs are known to act as complementary, antidotal or inimical to a particular drug. Practitioners can follow this rule when they apply one drug following another. Potentized Nux vomica can reduce acute hypnotic effect of alcohol on toads. Sulphur and Sepia are reported to be complementary to Nux-vom, while Coffea cruda and Zincum met are antidotal and inimical to Nux, respectively [...] Methods: Five batches of toads, each comprising 20 individuals, were treated by partial immersion in a drug diluted with distilled water 1:500 for 20 min. The control consisted of 90% ethanol diluted with distilled water 1:500. The drugs were Nux vom 200 CH, Sulphur 200 CH, Sepia 200 CH, Coffea 200 CH and Zincum 200 CH. Toads of each batch were separately exposed to 260mM ethanol solution and tested every 10 min to see if they had lost their righting reflex (RR)[...] Results: Toads treated with the five drugs took significantly longer time (P<0.01, one-way ANOVA) to lose RR than those treated with the control. The time taken to lose RR was significantly longer (P<0.01, one-way ANOVA) with Sulphur 200 CH, Sepia 200 CH, Coffea 200 CH and Zincum 200 CH than with Nux vom 200 CH alone. The situation was same when Nux treatment was followed by each of the four drugs. Of the five drugs Coffea showed the strongest anti-hypnotic effect. Conclusion: 1. Drugs complementary, antidotal and inimical to Nux vom showed the same anti-alcoholic effect as Nux in terms of increased tolerance to alcohol anesthesia. 2. The anti-alcoholic effect of Nux vom was markedly superseded by the above four drugs. 3. Of all the drugs tested Coffea showed the strongest anti-alcoholic effect. 4. It appears that the above four drugs produced their individual dominant effect cancelling the individual effect of Nux vom.