Extraction, identification and antitumor activity in cell cultures of cubebin isolated from dry seeds Piper cubeba (Piperaceae).

The Pipercubeba, it is one spice, widely consumed in Europe, which has several bioactive molecules, between those a lignan named cubebin. Cubebin has several known biological activities, such as analgesic activity and anti-inflammatory, trypanocidal activity, leishmanicidal and antitumor activity. The objective of this study was to evaluate the antiproliferative activity "in vitro" cubebin in eight different human tumor cell lines. It was fully characterized by IR analysis, NMR, mass spectrometry, DSC, TGA, residual solvent and elemental analysis. The antitumor activity of cubebin was evaluated "in vitro" on eight different human tumor cell lineages. Cubebin showed GI50≤30µg/mL for lineage cell U251 (glioma CNS), 786-0 (kidney), PC-3 (prostate), HT-29 (colon rectum). For K562 cells (leukemia), cubebin presented GI50≤to 4.0mg/mL. For the other lineages cells, MCF-7 (breast) and NCI-H460 to cubebin can be considered inactive because of GI50>250mg/mL. Analyzing the selectivity index for cubebin, it can be observed that high selectivity of cubebin to K562 lineage cells (leukemia). Analyzing the cytotoxic potential of cubebin was observed that probably acts cubebin altering metabolism, inhibiting cell growth - a cytostatic effect, showing no cytocidal effect on any lineage cell.

Pracaxi oil affects xenobiotic metabolisms, cellular proliferation, and oxidative stress without cytotogenotoxic effects in HepG2/C3A cells.

Pentaclethra macroloba (Willd.) Kuntze seeds oil has been used as a topical healing agent, applied mainly to parturients and snake bites. The objective was to investigate the effects of pracaxi oil (POP) on HepG2/C3A cells under cytogenotoxicity, cell cycle and apoptosis influence, and expression of metabolism and other related cell types proliferation genes. Cytotoxicity was analyzed by MTT test and apoptosis and cell cycle interferences by flow cytometry. To identify genotoxicity were used comet and micronucleus tests. RT-qPCR investigated gene expression. PO chemical characterization has shown two significant triterpenes, identified as oleanolic acid and hederagenin. The results showed that the PO did not reduce cell viability at concentrations ranging from 31 to 500 µg/ml. Comet and micronucleus assays revealed the absence of genotoxic effects, and flow cytometry showed no cell cycle or apoptosis disturbance. RT-qPCR indicated that PO up-regulated genes related to metabolism (CYP3A4, CYP1A2, CYP1A1), cell proliferation (mTOR), and oxidative stress (GPX1). The data indicate that PO has no cytogenotoxic effects and suggest that it activated the PI3/AKT/mTOR cascade of cell growth and proliferation. Inside the cells, the PO activated xenobiotic metabolizing genes, responsible for reactive oxygen species (ROS) generation, can neutralize ROS with increased GPX1 gene expression without genetic damage, interruption of the cell cycle, or induction of apoptosis.

In vivo toxicogenic potential of Salix alba (Salicaceae) bark extract.

Salix alba (white willow) bark extract is widely used for conditions associated with inflammation, fever, microbial infection or pain. Exposure of human cultured leukocytes to S. alba in vitro noted a genotoxic response. However, data regarding the influence of this bark extract on DNA damage in vivo are lacking. The main goal of this study was to examine the potential of S.alba bark extract to induce DNA damage and chromosome aberrations in an in vivo model using cells obtained from male Swiss albino mice administered the compound orally. The extract was administered by oral gavage daily for 7 days at doses of 500, 1000, or 2000 mg/kg b.w. Genotoxicity analysis was performed using the comet assay on peripheral blood leukocytes, as well as liver, bone marrow, heart, and testicular cells collected 4 hr after the last treatment and the micronucleus (MN) test on bone marrow cells. In essence cells were collected 28 hr after the penultimate treatment Data demonstrated that S. alba bark extract did not induce significant DNA damage in any cell types examined, or clastogenic/aneugenic effects as detected by the MN test at the three tested doses. Under these experimental conditions, evidence indicates that S.alba bark extract did not initiate genotoxic or chromosome aberrations in various mouse cells investigated.

Genotoxicity induced by nerol, an essential oil present in citric plants using human peripheral blood mononuclear cells (PBMC) and HepG2/C3A cells as a model.

Nerol (cis-3,7-dimethyl-2,6-octadien-1-ol) is a monoterpene widely used in cosmetic products, household detergents and cleaners, as well as a flavoring in several food products. Despite the high level of human exposure to nerol, an absence of studies regarding potential genetic toxicity in human cells exists. The aim of this investigation was to examine the cytotoxic and genotoxic potential of this monoterpene on human peripheral blood mononuclear cells as well as hepatic metabolizing HepG2/C3A human cell line. Cytotoxicity was assessed using trypan blue staining and MTT assay while genotoxicity was determined utilizing the comet and micronucleus test. Cytotoxicity tests showed cell viability greater than 70% for concentrations between 2.5 and 500 µg/ml. Both cell types exhibited significant DNA damage and chromosomal mutations after medium and high concentration incubation with nerol indicating that the safety of use of this monoterpene in various formulations to which humans are exposed needs to be monitored and requires more comprehensive investigations.

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.

Salix alba (white willow) medicinal plant presents genotoxic effects in human cultured leukocytes.

Salix alba (SA), commonly known as white willow, is a plant used in folk medicine for the treatment of chronic and acute inflammation, infection, pain, and fever. The phytochemical characterization of the bark extract of this plant indicated that its main component is salicin, a precursor of the anti-inflammatory agent acetylsalicylic acid. Considering the lack of studies evaluating the genetic toxicity and cytotoxic action of SA bark extract on human cells, as well as the chemical characterization of its major phenolic compounds, the present study was designed to (1) investigate the cytotoxic and genotoxic potential of SA bark extract on human peripheral leukocyte cells and human hepatoma cell line HepG2, and (2) characterize its major phenolic constituents. The phenolic compounds found were salicylic acid, salicin, salidroside, saligenin, tremulodin, salicoylsalicin, salicortin, and tremulacin. The results using trypan blue staining test showed viability decreases (viability less than 70%) for concentrations of SA extract equal and higher to 200 µg/ml. Low genotoxic activity (comet assay) was exhibited for 50 and 100 µg/ml SA extract in human leukocytes. SA did not exert a marked clastogenic/aneugenic effect on leukocytes and HepG2 human cells. Data suggest that the genotoxic effects of SA bark extract occur when it is not metabolized by liver enzymes.

The genotoxic effects of fruit extract of Crataegus oxyacantha (hawthorn) in mice.

Crataegus oxyacantha L. (Rosaceae) is a medicinal plant with a long history of use in European, Chinese, and American. The majority of pharmacological activities associated with fruit extracts of C. oxyacantha L. are related to cardio-stimulant properties utilized in the treatment of atherosclerosis, hypertension with myocardic insufficiency, angina pectoris, cardiac rhythm alterations, and heart failure. Some other therapeutic uses for renal calculi, dyspnea, as well as a diuretic, sedative, and anxiolytic were also reported. Due to the beneficial potential of C. oxyacantha fruits extract but evidence in vitro of genetic toxicity, the aim of the present study was to examine the genotoxic potential of plant extract in vivo in mice. The extract was administered orally, daily by gavage at doses of 50, 100, and 200 mg/kg body weight for seven days. Data demonstrated that C. oxyacantha extract did not markedly induce DNA damage in leukocytes and bone marrow cells by the comet assay; however, the extract produced a significant rise in micronucleated polychromatic erythrocytes (PCE) at all tested doses in a non-dose dependent manner as evidenced by the micronucleus test. The PCE/normochromatic erythrocytes (NCE) ratio indicated no significant cytotoxicity. Under our experimental conditions, C. oxyacantha fruits extract exhibited weak clastogenic and/or aneugenic effects in bone marrow cells of male mice, confirming our previous in vitro findings that this plant extract induced genotoxicity suggesting that prolonged or high dose use needs to be undertaken with caution.

Fruit extract of the medicinal plant Crataegus oxyacantha exerts genotoxic and mutagenic effects in cultured cells.

Crataegus oxyacantha, a plant of the Rosaceae family also known "English hawthorn, haw, maybush, or whitethorn," has long been used for medicinal purposes such as digestive disorders, hyperlipidemia, dyspnea, inducing diuresis, and preventing kidney stones. However, the predominant use of this plant has been to treat cardiovascular disorders. Due to a lack of studies on the genotoxicity of C. oxyacantha, this investigation was undertaken to determine whether its fruit extract exerts cytotoxic, genotoxic, or clastogenic/aneugenic effects in leukocytes and HepG2 (liver hepatocellular carcinoma) cultured human cells, or mutagenic effects in TA100 and TA98 strains of Salmonella typhimurium bacterium. Genotoxicity analysis showed that the extract produced no marked genotoxic effects at concentrations of 2.5 or 5 µg/ml in either cell type; however, at concentrations of 10 µg/ml or higher significant DNA damage was detected. The micronucleus test also demonstrated that concentrations of 10 µg/ml or higher produced clastogenic/aneugenic responses. In the Ames test, the extract induced mutagenic effects in TA98 strain of S. typhimurium with metabolic activation at all tested concentrations (2.5 to 500 µg/ml). Data indicate that, under certain experimental conditions, the fruit extract of C. oxyacantha exerts genotoxic and clastogenic/aneugenic effects in cultured human cells, and with metabolism mutagenicity occurs in bacteria cells.

Hematological and hepatic effects of vascular epidermal growth factor (VEGF) used to stimulate hair growth in an animal model.

BACKGROUND: Alopecia areata is the hair loss usually reversible, in sharply defined areas. The treatment of alopecia using growth factors shows interesting activity in promoting hair growth. In this concept, VEGF (vascular endothelial growth factor) is a marker of angiogenesis, stimulating hair growth by facilitating the supply of nutrients to the hair follicle, increasing follicular diameter. The aim of this study was the evaluation of a topical gel enriched with VEGF liposomes on the hair growth stimulation and its toxicological aspects. METHODS: Mesocricetus auratus were randomly divided into three groups. Control group was treated with Aristoflex® gel, 1% group with the same gel but added 1% VEGF and 3% group with 3% VEGF. Biochemical, hematological and histological analyses were done. RESULTS: At the end of the experiment (15th day of VEGF treatment) efficacy was determined macroscopically by hair density dermatoscopy analysis, and microscopically by hair diameter analysis. They both demonstrated that hair of the VEGF group increased faster and thicker than control. On the other hand, biochemical and hematological results had shown that VEGF was not 100% inert. CONCLUSIONS: VEGF increased hair follicle area, but more studies are necessary to confirm its toxicity.

Evaluation of antimicrobial and antimalarial activities of crude extract, fractions and 4-nerolidylcathecol from the aerial parts of Piper umbellata L. (Piperaceae).

Presently, natural products, such as Piper umbellata L. (Piperaceae), have been evaluated as sources of antimicrobial agents with efficacies against microorganisms. The in vitro antimicrobial activity was performed by broth micro-dilution susceptibility assay, according to the protocols of the National Committee for Clinical Laboratory Standards, and described the antibacterial and antifungal activities of crude ethanolic extract and fractions obtained by partitions sequentially among water-methanol, methylene chloride and ethyl acetate, as well as the major constituent, 4-nerolidylcatechol from the aerial parts of P. umbellata L. Amphotericin B and ciprofloxacin were used as controls. Among the microorganism cultures, hydromethanol fraction demonstrated the pre-eminent antifungal activity. 4-Nerolidylcathecol was the only tested plant component that exhibited activity against all the selected microorganisms, suggesting its great potential as a source for the development of new drugs. In order to estimate the antimalarial activity of P. umbellata L., a micro-dilution method protocol, parasite lactate dehydrogenase assay, with a Plasmodium falciparum Sierra Leone (D6) clone was utilised. The antimalarial agent artemisinin was used as control. 4-Nerolidylcathecol exhibited the best antimalarial activity; however, it was not significant when compared with control. These in vitro results do not justify the use of P. umbellata L. in malaria patients. However, there is a possibility of 4-nerolidylcathecol, after biotransformation, exhibiting a significant antimalarial activity in in vivo assays. However, 4-nerolidylcathecol demonstrated to possess a broad antimicrobial activity which is, in fact, a promising source for the development of new therapeutic agents.

Dragon's blood Croton palanostigma induces genotoxic effects in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Dragon's blood is a dark-red sap produced by species from the genus Croton (Euphorbiaceae), which has been used as a famous traditional medicine since ancient times in many countries, with scarce data about its safe use in humans. In this research, we studied genotoxicity and clastogenicity of Croton palanostigma sap using the comet assay and micronucleus test in cells of mice submitted to acute treatment. MATERIAL AND METHODS: HPLC analysis was performed to identify the main components of the sap. The sap was administered by oral gavage at doses of 300 mg/kg, 1,000 mg/kg and 2,000 mg/kg. For the analysis, the comet assay was performed on the leukocytes and liver cells collected 24h after treatment, and the micronucleus test (MN) on bone marrow cells. Cytotoxicity was assessed by scoring 200 consecutive polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio). RESULTS AND CONCLUSION: The alkaloid taspine was the main compound indentified in the crude sap of Croton palanostigma. The results of the genotoxicity assessment show that all sap doses tested produced genotoxic effects in leukocytes and liver cells and also produced clastogenic/aneugenic effects in bone marrow cells of mice at the two higher doses tested. The PCE/NCE ratio indicated no cytotoxicity. The data obtained suggest caution in the use of Croton palanostigma sap by humans considering its risk of carcinogenesis.

Genotoxicity assessment of Garcinia achachairu Rusby (Clusiaceae) extract in mammalian cells in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: Garcinia achachairu Rusby (Clusiaceae) is popularly known as "achachairu", and is used in Bolivian folk medicine for its healing, digestive, and laxative properties, and in the treatment of gastritis, rheumatism and inflammation. Despite its widespread therapeutic use, there is a lack of data regarding its in vivo genotoxic effects. Therefore, in this study, we used the comet assay and the micronucleus test, respectively, to evaluate the possible genotoxic and clastogenic effects of Garcinia achachairu seed extract (GAE) on different cells of mice. MATERIAL AND METHODS: The GAE was administered by oral gavage at doses of 500, 1000 and 2000 mg/kg. For the analysis, the comet assay was performed on the leukocytes (collected 4 and 24 h after treatment), liver, bone marrow and testicular cells (collected 24 h after treatment), and the micronucleus test (MN) on bone marrow cells. Cytotoxicity was assessed by scoring 200 consecutive polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio). RESULTS AND CONCLUSION: The results showed that GAE did not induce significant DNA damage in leukocytes (4 h and 24 h samples), liver, bone marrow and testicular cells (24 h samples). GAE also did not show any significant increase in micronucleated polychromatic erythrocytes (MNPCEs) at the three tested doses. The PCE/NCE ratio indicated no cytotoxicity. Under our experimental conditions, the data obtained suggest that a single oral administration of G. achachairu extract does not cause genotoxicity and clastogenicity in different cells of mice.

Genotoxicity assessment of the antimalarial compound artesunate in somatic cells of mice.

Artesunate is a derivate of artemisinin that is both an antimalarial agent and acts cytotoxically on tumor cells. Despite its therapeutic use, its in vivo genotoxic potential has still not been evaluated. This study, therefore, was an investigation into the effects of a single oral administration of artesunate with an in vivo comet assay that analyzed leukocytes from peripheral blood and liver cells, and a micronucleus (MN) assay of bone marrow cells from male Swiss mice. The artesunate was administered by oral gavage at doses of 5, 50 and 100 mg/kg. Cytotoxicity was assessed by scoring 200 consecutive polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio). The results demonstrate that artesunate induced significant DNA damage only in liver cells and that high doses of artesunate caused an increase in the mean number of micronucleated polychromatic erythrocytes (MNPCE). Under our experimental conditions, artesunate showed weak genotoxic effects at low doses and clastogenic effects at high doses. The PCE/NCE ratio indicated no cytotoxicity. The data obtained suggest caution about either continuous or high-dose use of artesunate by humans.

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.(AU)

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.(AU)

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.(AU)

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...

Genotoxic effects of (-)-cubebin in somatic cells of mice.

(-)-Cubebin belongs to the dibenzylbutyrolactone lignan group, which is widely distributed in the plant kingdom. Because this compound shows interesting biological activities, it is extremely important to evaluate its possible genotoxic activity to allow its safe use in humans. Thus, the present study was performed to investigate the genotoxicity potential activity of (-)-cubebin assessed by two assays: micronucleus in bone marrow cells and comet test in peripheral blood leukocytes of Swiss mice. In the (-)-cubebin dose range-finding assays, the maximum tolerated dose was greater than 2000 mg kg(-1) . The compound was administered by an oral route at single doses of 250, 500 and 2000 mg kg(-1) body weight. Cytotoxicity was assessed by scoring 200 consecutive total polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio). Under our experimental conditions, micronucleus and comet assays, respectively, showed that (-)-cubebin caused dose-related clastogenic and genotoxic effects in the somatic cells investigated. PCE/NCE ratio showed no cytotoxicity for the three doses of the compound. The data suggest caution in the ingestion of (-)-cubebin by humans, especially at high doses.

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.

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.(AU)

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.(AU)