Cytotoxic effects of Euterpe oleraceae fruit oil (açaí) in rat liver and thyroid tissues

Abstract Euterpe oleracea Mart., Arecaceae, fruit (açaí) presents considerable potential for the development of new medicines due to its phytochemical composition and antioxidant activity. More recently, special attention has been given to the pharmacological potential of the fruit's oil. This study analysed the histological and histochemical effects of different dosages of açaí oil on rat's liver and thyroid cells, in order to evaluate its cytotoxic potential after administration for consecutive days. Male Wistar rats were treated with the açaí oil by gavage at doses of 30, 100 and 300 mg/kg, for 14 days, within a 24 h interval. Liver and thyroid fragments were collected for histology (hematoxylin and eosin) and histochemistry analysis (blue of Nilo (lipids), Baker (lipids), bromophenol blue (protein), PAS (polysaccharides)). The results showed that animals exposed to açaí oil presented alterations in the liver cells, where the integrity of the liver tissue was increasingly lost as the açaí oil doses increased. Nuclear pyknosis was observed in several hepatocytes, evidencing the occurrence of cell death. Alteration in the amount of lipids, polysaccharides, vacuoles in the cytoplasm, and proliferation of Kupffer cells were observed in histochemical analyzes. As for the thyroid of the treated rats, alterations were observed in the size of the follicular lumen and also in the connective tissue found between the follicles. Under the experimental conditions employed in the present study, the cytotoxicity observed in this work is worrying, specially considering the liver, when frequent or continuous damage could lead pathological disorders in this organ.

Evaluation and study of mebendazole polymorphs present in raw materials and tablets available in the Brazilian pharmaceutical market.

The dissolution of a drug can be compromised by the presence of different polymorphs, which may have different solubilities. Importantly, the pharmacopoeiamonographs,usually not have tests for the characterization of these polymorphic forms of a drug. Was performed a study of polymorphic forms of mebendazole present in raw materials and also pills available in the Brazilian pharmaceutical market through the techniques of infrared (FTIR), differential scanning calorimetry (DSC), dissolution , solubility and X-ray diffraction pattern (XRPD). Through the analysis of FTIR and DSC curves showed that there are three main polymorphic forms of mebendazole present in raw materials and tablets that compound. The data obtained in the dissolution and solubility tests showed that Form A is less soluble than Form B which is less soluble than the C form, when using a dissolution medium without added surfactant. It has been found that in some tablets mebendazole there is a mixture of polymorphic forms, and that the raw materials present two major polymorphic forms. Then it is suggested the need of quality control regarding the type of polymorph used in the production of mebendazole tablets to ensure greater therapeutic efficacy.

Development and Validation of RP-HPLC Method for the Determination of Related Compounds in Quetiapine Hemifumarate Raw Material and Tablets.

A simple and sensitive high performance liquid chromatographic method has been developed for the determination of assay quantitative of related compounds and quetiapine hemifumarate in raw material and tablets. Quetiapine hemifumarate is used for the treatment of schizophrenia and there are some generic medicines available in brazilian marketing pharmaceutical, it´s necessary evaluate the quality control of raw material used in the production. Efficient chromatographic separation was carry out on a C18 stationary phase with mobile phase consisting in a mixture of phosphate buffer pH 6.6:Acetonitrile:Methanol (45:40:15), flow rate of 1.0 mL min-1, injection volume of 20 μL, temperature of 25 °C and ultraviolet detection at 220 nm. All of the chromatographic parameters were attended, with resolution greater than 2.9 between quetiapine hemifumarate and impurities. The HPLC method was validated according ICH guidelines, evaluating selectivity, limits of detection and quantification, linearity, accuracy, precision and robustness. The relative retentions times were about 0.58, 0.69 and 0.88 to related compounds, piperazine, lactam and ethanol compound, respectively. Impurities were found < 0.1 % in samples and the assay of quetiapine hemifumarate was > 98.15%. The method can be used for the routine analysis of the impurities in Quetiapine hemihumarate (QH) without any interference.

Gingko biloba leaves extract on growth and morphology of trypanosomatids / Efecto del extracto de las hojas de Gingko biloba en el crecimiento y la morfología de tripanosomátidos

Gingko biloba has been one of the most used medicinal plants all over the world in the past years. In this study, our group has studied the effect of a hydroethanolic extract from the aerial parts of this plant on the growth and morphological differentiation of trypanosomatids. Herpetomonas samuelpessoai and Herpetomonas sp were used in this study. The extract was obtained in a Soxhlet apparatus (50 oC, 2 hours). This extract was aseptically added to Roitman’s medium in different concentrations (4, 20, 40, 60, 80 and 100 mg/ml). The growth rate was determined using a Newbauer chamber to count numbers of cells after the extract inoculation (24 and 72 hours later). Smears stained by the Panotic method was used to determine the percentages of pro, para and opisthomastigote forms. The extract inhibited Herpetomonas sp growth in concentrations higher than 20 mg/ml. H. samuelpessoai has been inhibited in doses higher than 40 mg/ml. No morphological differentiation was observed in Herpetomonas sp cell. However, morphological differentiations could be noticed in H. samuelpessoai cell using doses higher than 40 mg/ml. These alterations are probably related to the cell division process, since cells with 3 or 4 nucleus were observed. Also, cytoplasmatic expansions, representing unsuccessful process of cell division were frequently found out. Further ultrastructural analysis using a transmission electron microscope showed cells with homogeneous nucleus or the absence of it. Protozoan protein profile was also analyzed. It was possible to notice changes in both trypanosomatids used in this study. H. samuelpessoai has shown over expression and accumulation of proteins which its degradation is essential to continue the cell differentiation. Also, it is possible to suggest that this extract acts through the modulation of the genetic expression and may be harmful to human cells if not purified.

Gingko biloba es una de las plantas medicinales más utilizadas en todo el mundo en los últimos años. En este estudio, nuestro grupo ha estudiado el efecto de un extracto hidroetanólico de la parte aérea de esta planta sobre el crecimiento y la diferenciación morfológica de tripanosomátidos. Herpetomonas samuelpessoai y Herpetomonas sp se utilizaron en este estudio. El extracto se obtuvo en un aparato Soxhlet (50° C/2 horas). Este extracto se agregó asépticamente a medio Roitman en diferentes concentraciones (4, 20, 40, 60, 80 y 100 mg /ml). La tasa de crecimiento se determinó utilizando una cámara de Newbauer para contar el número de células después de la inoculación de extracto (24 y 72 horas más tarde). Frotis teñidos por el método Panotic se utilizó para determinar los porcentajes de pro, para y las formas opistomastigota. El extracto inhibió el crecimiento Herpetomonas sp en concentraciones superiores a 20 mg /ml. H. samuelpessoai se ha inhibido en dosis superiores a 40 mg /ml. No se observó diferenciación morfológica en la celda Herpetomonas sp. Sin embargo, las diferenciaciones morfológicas se pudo observar en la celda H. samuelpessoai con dosis superiores a 40 mg /ml. Estas alteraciones son probablemente relacionado con el proceso de división celular, ya que las células con 3 o 4 núcleos se observaron. Además, las expansiones citoplasmáticas, lo que representa el proceso fallido de la división celular se encontraron con frecuencia hacia fuera. Un análisis más detallado ultraestructural usando microscopio electrónico de transmisión mostró células con núcleo homogéneo o la ausencia de ella. El perfil de proteínas por Protozoarios también se ha analizado. Fue posible notar cambios tanto en tripanosomátidos utilizados en este estudio. H. samuelpessoai ha demostrado a lo largo de expresión y la acumulación de proteínas que su degradación es esencial para continuar con la diferenciación celular. Además, es posible sugerir que este extracto...

Anxiolytic and antidepressive effects of the homeopathic complex Homeo-pax® (pre-clinical study) / Anxiolytic and antidepressive effects of the homeopathic complex Homeo-pax® (pre-clinical study)

The homeopathic complex Homeo-Pax® has been used as an antidepressant and anxiolytic homeopathic medicine available in Brazil. It is a complex mixture prepared with Aconitum nap.6cH, Aurum met. 6cH, Phosphorus 6cH, Argentum nitricum 6cH, Arsenicum alb. 6cH, and Valeriana officinalis 3cH. This study had evaluated the behavior in rats after treatment with Homeo-Pax® in pre-clinical models of depression and anxiety. Elevated Plus Maze Test (EPM), Forced Swimming Test (FST), Open Field Test (OFT) and the Rota Rod Test (RRT) behavior assays were used to confirm its activity. In the EPM, the animals treated with Homeo-pax® on the 1st day and until the 20th day of treatment remained longer in the open arms of the maze than on 30th day. This result was statistically significant compared with the control group (p < 0.05). In the FST, the treatment with Homeo-pax® (0.5 ml, p.o) increased the swimming time, compared to the control group. This effect was dependent on treatment time, resulting in a similar effect to that presented by amfepramone (10 mg/kg, p.o). In the OFT, crossing by the animals was significantly increased by the treatment with amfepramone (10mg/kg, p.o), and also with the 30- day treatment with Homeo-pax® . In the RRT, the 30-day treatment with Homeo-pax® (0.5 ml, p.o) did not affect the animals? motor coordination, compared with the control group, which presented the same behavior. Based on the results obtained, it can be suggested that the homeopathic complex Homeo-pax® has anxiolytic and antidepressant properties without affecting motor coordination capacity.

O complexo homeopático Homeo-Pax® tem sido usado no Brasil como um medicamento homeopático de ação antidepressiva e ansiolítica. O Homeo-Pax® é um complexo preparado com Aconitum nap. 6cH, Aurum met. 6cH, Phosphorus 6cH, Argentum nitricum 6cH, Arsenicum alb. 6cH e Valeriana officinalis 3cH. Este estudo avaliou o comportamento de ratos após o tratamento com Homeo-Pax® em modelos pré-clínicos de depressão e ansiedade. Testes de labirinto em cruz elevado (EPM), nado forçado (FST), campo aberto (OFT) e Rotarod (RRT) foram usados para avaliar a atividade dos animais. No EPM, os animais tratados com Homeo-pax® permaneceram mais tempo nos braços abertos do labirinto, durante do 20 primeiros dias de tratamento, em relação ao 30º dia. Este resultado foi estatisticamente significativo quando comparado com o grupo controle (p < 0.05). No FST, o tratamento com Homeo-pax® (0.5 ml, p.o) aumentou o tempo de nado, comparado ao grupo controle. Este efeito foi dependente o tempo de tratamento, resultando similar ao efeito da amfepramona (10 mg/kg, p.o). No OFT, o movimento dos animais foi significativamente aumentado pelo tratamento com amfepramona (10mg/kg, p.o) e também no 30º dia de tratamento com Homeo-pax® . No RRT, o tratamento por 30 dias com Homeo-pax® (0.5 ml, p.o) não afetou a coordenação motora dos animais, em relação ao grupo controle. Baseado nesses resultados, pode ser sugerido que o complexo homoepático Homeo-pax® tem propriedades ansiolíticas e antidepressivas sem afetar a coordenação motora.

El complejo homeopático Homeo-pax® viene siendo usado en Brasil como un medicamento homeopático de acción antidepresiva y ansiolítica. El Homeo-pax® es un complejo preparado con Aconitum nap 6cH, Aurum Met 6cH, Phosphorus 6cH, Argentum Nitricum 6cH, Arsenicum Alb 6cH y Valeriana officinalis 3cH. Este estudio evaluó el comportamiento de camondongos después del tratamiento con Homeo-pax® en modelos preclinicos de depresión y ansiedad. Testes de laberinto en cruz elevado (EPM) nado forzado (FST), campo abierto (OFT) y Rotarod (RRT) fueron usados para evaluar la actividad de los animales. En el EPM los animales tratados com Homeo-pax® permanecieron mas tiempo en los brazos abiertos del laberinto durante los 20 primeros dias de tratamiento en relación al 30º dia. Este resultado fue estadísticamente significativo si comparado con el grupo control (p<0.05). En el FST, el tratamiento con Homeo-pax® (0.5 ml,p.o) aumentó el tiempo de nado, comparado al grupo control. Este efecto fue dependiente del tiempo de tratamiento, resultando similar al efecto de la anfepramona (10 mg/kg, p.o). En el OFT, el movimiento de los animales fue significativamente aumentado por el tratamiento con anfepramona (10mg/kg, p.o) y tambien en el 30º dia de tratamiento con Homeo-pax® . En el RRT el tratamiento por 30 dias con Homeo-pax® (0.5 ml, p.o) no afectó la coodinación motora de los animales, en relación al grupo control. Basado en esos resultados puede ser sugerido que el complejo homeopático Homeo-pax® tiene propiedades ansiolíticas y antidepresivas sin afectar la coordinación motora.

Study of high dilutions of copaiba oil on inflammatory process / Estudo de altas diluições homeopáticas do óleo de copaíba

The discovery of new drugs has led to a need to develop techniques to control the occurrence of toxic and collateral effects. This has enabled the advancement of homeopathic therapeutics as it presents major advantages against these effects. This study was designed to explore the effects of high dilutions of Copaifera (copaiba oil) on inflammation. This study considered the way the high dilutions were obtained (triturated form or mother-tincture-MT). The preparations were administered orally. The effects of the dilutions were tested using the rat paw edema induced by carrageenan; granuloumatous tissue induction and the edema induced by Croton oil. The high dilutions of copaiba oil obtained from both trituration and MT produced a statistically significant inhibitory effect of the carrageenan edematogenic process compared to control. The maximum effect was observed with dilution 30cH, with inhibition of edema by 73%, whereas indomethacin was 55%. Subcutaneous implantation of cotton pellets have induced a granulomatous tissue, evaluated 7 days after implantation. Daily treatment with dexamethasone produced 53% inhibition on the formation of granulomatous tissue. The 6cH dilution of copaiba oil inhibited in a statistically significant way the formation of granulomatous tissue compared to the control (18% and 16%, respectively). Edema in Croton-oil induced dermatitis was intense. Groups treated with dexamethasone and dilutions of copaiba oil presented similar responses, with inhibition by 57% and 48% respectively. Based on the results obtained in this study, it may be suggest that the Copaiba oil high dilutions possess an anti-inflammatory property supporting its use in the treatment of inflammatory disorders.

A descoberta de novas drogas tem gerado a necessidade de desenvolvimento de novas técnicas para controle da ocorrência de efeitos tóxicos e colaterais. Isto tem favorecido o uso da terapêutica homeopática uma vez que esta apresenta vantagens contra alguns efeitos adversos. Este trabalho foi proposto visando explorar os efeitos de Altas Diluições de Copaifera (óleo de copaíba) sobre inflamação. Foi considerada a maneira como as altas diluições foram obtidas (via trituração ou via tintura-mãe). As preparações foram administradas oralmente. Os efeitos das diluições foram avaliados usando um modelo de edema em pata de ratos, induzido por carragenina, indução de tecido granulomatoso e edema induzido pelo óleo de Croton. As altas diluições de óleo de Copaíba obtidas a partir de trituração e de tintura-mãe produziram efeitos inibitórios estatisticamente significativos para o processo endematogênico, quando comparado com o controle. O efeito máximo foi obtido com a diluição 30cH, com uma inibição de 73% do edema, enquanto a indometacina inibiu 55%. O implante subcutâneo de pellets de algodão induziram a granulomatose do tecido, avaliados 7 dias após o implante. Tratamento diário com dexametasona produziu 53% de inibição da formação de tecido granulomatoso. A diluição 6cH do óleo de copaíba inibiu de forma significativa a formação de tecido granulomatoso, comparado com o controle (18% e 16% respectivamente). Edema em dermatite induzida por óleo de Croton foi intensa. Grupos tratados com dexametasona e diluições de óleo de copaíba apresentaram respostas similares, com inibição de 57% e 48% respectivamente. Baseado nos resultados obtidos, pode-se concluir que altas diluições do óleo de copaíba apresentam efeitos anti-inflamatórios, sugerindo seu uso no tratamento de desordens inflamatórias.

Clastogenicity of Piper cubeba (Piperaceae) seed extract in an in vivo mammalian cell system

The plant Piper cubeba is widely distributed in tropical and subtropical regions and is used medically for various purposes but has not yet been evaluated for genotoxicity. We used male and female Swiss mice and Wistar rats and the comet assay and micronucleus test to investigate the mutagenic potential of a crude extract of P. cubeba seeds. The rodents were administered 0.5 g kg-1, 1.0 g kg-1 and 1.5 g kg-1 of the extract by gavage. For the Swiss mice, peripheral blood was collected 24 h after treatment for the comet assay, and at 48 and 72 h for the micronucleus test. For the Wistar rats, peripheral blood and hepatic cells were collected for the comet assay and bone marrow cells were collected for the micronucleus test 24 h after treatment. At 1.5 g kg-1, the highest dose tested, the extract induced a statistically significant increase in both the mean number of micronucleated polychromatic erythrocytes and the level of DNA damage in the rodent cell types analyzed. Under our experimental conditions, the P. cubeba seed extract was genotoxic in vivo when administered orally to mice and rats.

Effect of betamethasone and diclofenac sodium on serum and tissue concentration of amoxicillin: in vivo study in rats%Efeito da betametasona e do diclofenaco sódico na concentração sérica e tecidual da amoxicilina: estudo in vivo em ratos

OBJETIVO: A prescrição de antimicrobianos associados a antiinflamatórios é uma prática comum em odontologia, embora na maioria das vezes não haja justificativa para tal conduta. O objetivo deste trabalho foi avaliar, em um estudo in vivo em ratos, os efeitos da betametasona e do diclofenaco sódico nas concentrações sérica e tecidual da amoxicilina. MÉTODOS: Foram utilizados 48 ratos Wistar machos (6 grupos, n=8), com idade de 60 dias. Esponjas de PVC (policlorovinil) foram implantadas em quatro pontos no dorso de cada animal. Após sete dias, foram administrados por via intragástrica ou intramuscular: Grupo 1: amoxicilina (25 mg/kg); G2: diclofenaco sódico (2,5 mg/kg/i.m.); G3: betametasona (0,1 mg/kg/i.m.); G4: diclofenaco sódico e amoxicilina; G5: betametasona e amoxicilina; e G6: solução de cloreto de sódio a 0,9% (1,0 mL - grupo controle). Após 90 minutos, foram colhidos 2 tecidos granulomatosos e amostras séricas de cada animal e colocados em meios de cultura inoculados com 108 ufc/mL de Staphylococcus aureus ATCC 25923. Os halos de inibição foram medidos após 18 horas de incubação (37ºC), e através do teste microbiológico foram obtidas as concentrações séricas e teciduais da amoxicilina. RESULTADOS: Não foram observados halos de inibição para os grupos 2, 3 e 6. As concentrações séricas e teciduais de G1 (4,14µg/g e 2,06µg/mL, respectivamente) e G5 (3,87µg/g e 1,70µg/mL, respectivamente) demonstraram diferenças estatisticamente significantes (Kruskal-Wallis, p<0,05) em comparação a G4 (1,45µg/g e 0,41µg/mL, respectivamente). G1 e G5 não apresentaram diferença estatística (p>0,05). CONCLUSÃO: Considerando uma dose única, a betametasona não interferiu nas concentrações sérica e tecidual de amoxicilina, enquanto o diclofenaco sódico reduziu as concentrações sérica e tecidual de amoxicilina em ratos..

Avaliaçäo do potencial alergênico de um novo produto fermentado de soja / Evaluation of allergenic potential of a novel fermented soymilk product

O processo fermentativo induz uma reduçäo no pH do meio e, dependendo do cultivo bacteriano, observa-se uma açäo proteolítica com diminuiçäo da alergenicidade presente em algumas fraçöes protéicas da soja. No presente estudo, o objetivo foi avaliar o potencial alergênico induzido pela ingestäo oral de um novo produto fermentado de soja, buscando demonstrar a relaçäo existente entre o processo fermentativo e a alergenicidade. Trinta ratos Wistar, machos, adultos, foram separados em cinco grupos: 1. animais tratados com produto fermentado; 2. animais tratados com o produto näo fermentado (placebo); 3. animais sensibilizados com ovalbumina (controle positivo); 4. animais sem nenhum tratamento (somente soluçäo de Tyrode; controle negativo) e 5. animais estimulados pela açäo direta do produto fermentado, placebo e soluçäo de Tyrode. Os produtos fermentados e placebo foram obtidos de acordo com Rossi, et al. No exsudato peritoneal foi determinado o percentual de liberaçäo de histamina e concomitantemente, no mesmo animal, foi verificada a degranulaçäo de mastócitos. Os resultados mostraram que o produto fermentado apresentou um percentual de liberaçäo de histamina (12,6 ñ 2,1 por cento) e degranulaçäo de mastócitos (2,92 ñ 0,9 por cento) menores que o observado para o placebo (18,95 ñ 3,2 por cento e 5,5 ñ 2,3 por cento, respectivamente). Concluiu-se que o produto fermentado näo apresentou potencial alergênico e que o processo fermentativo reduziu favoravelmente a alergenicidade e que o processo fermentativo reduziu favoravelmente a alergenicidade das proteinas de soja.