Repeated dose 90-day oral toxicity study of Vitex megapotamica (Spreng.) Moldenke.

Vitex megapotamica (Spreng.) Moldenke is a plant with medicinal properties popularly used in Brazil to treat diabetes and obesity. Despite the promising use of this plant, are still incipient toxicology studies on prolonged periods of treatment. The aim of this study was to evaluate the repeated dose 90-day oral toxicity study of V. megapotamica methanolic extract (VMME) in male and female Wistar rats. Different groups of rats (n = 8) were treated daily with three different doses of VMME (100, 300, and 900 mg/kg) or vehicle (filtered water). Body weight, water, and feed consumption, and clinical and behavioral changes were monitored daily. At the end of the experimental period, blood samples were obtained for hematological and biochemical analyzes. After euthanasia, the vital organs were removed for the determination of relative weight and for histopathological analysis. No clinical signs of toxicity or mortality were found during the experimental period. Treatment with VMME did not induce any change in body weight gain, eating patterns, and behavior. We found no statistically significant changes in the different hematological and biochemical parameters evaluated. The relative weight of the organs and histopathological analysis did not show any significant change when compared to animals treated with the vehicle. The data obtained in this study allow us to conclude that the VMME obtained from V. megapotamica is safe after a repeated-dose 90-day oral toxicity study in male and female Wistar rats.

Genotoxicity Study of Vitex megapotamica (Spreng.) Moldenke.

Vitex megapotamica (Spreng) Moldenke is commonly known as tarumã, it is an important medicinal and edible fruit plant. It is native to regions of tropical and subtropical climate in greater proportion than temperate zones and widely distributed in Central America, South America, Asia, and Africa. In Brazil, it is present in the Atlantic Forest and Cerrado biomes. Despite its widespread use, there are no minimum standards for quality control or information on genotoxicity. Therefore, the aim of this study was to present a detailed description of the short-term genotoxicity assays of V. megapotamica and to provide parameters of a preparation routinely used in traditional folk medicine. For genotoxicity assays, five groups were used with eight wistar rats in each group. For this, three doses of the V. megapotamica extract in doses (100, 300, and 900 mg/kg) or negative control (filtered water) were administered orally and positive control cyclophosphamide monohydrate (20 mg/kg; Sigma-Aldrich®) was applied by the intraperitoneal route after 24 h. At the end, whole blood was collected in a tube containing EDTA for the comet test and later the animals were euthanized. For the micronucleus test, femurs were removed, and bone marrow was collected. In the comet assay, V. megapotamica crude extract did not show significant DNA damage at all doses tested. The micronucleus assay showed no significant increase in the frequency of inducing micronuclei at any dose examined. It can be concluded that the safety parameters in genotoxicity studies reveal that V. megapotamica has no toxicity, which characterizes the important quality control of this plant species.

Safety assessment of MEFAS: an innovative hybrid salt of mefloquine and artesunate for malaria treatment.

Malaria is a global public health problem that causes approximately 445 000 deaths annually worldwide, especially in underdeveloped countries. Because of the high prevalence and mortality of the disease, new and less toxic therapeutic agents need to be developed, such as MEFAS, a low-cost hybrid salt that consists of artesunate and mefloquine. However, the efficacy of MEFAS has been systematically demonstrated, its safety requires further investigation. This study investigated the acute toxicity of MEFAS and its precursors, artesunate, and mefloquine. A total of 42 female Swiss mice were divided into seven groups (n = 6/group) that were treated orally by gavage with vehicle (filtered water, negative control), MEFAS (50, 500, and 1000 mg/kg), and 1:1 concentrations of artesunate + mefloquine (50, 500, and 1000 mg/kg). Clinical signs of toxicity were observed for 14 d after treatment. On day 15, the animals were weighed, deeply anesthetized with isoflurane, and euthanized for subsequent collection of the liver, spleen, and kidneys. The relative organ weights were determined, followed by histopathological analysis. Artesunate + mefloquine produced toxic effects compared with the negative control group, reflected by changes in clinical signs, relative organ weights, and histopathological alterations. In MEFAS-treated animals, no changes were observed compared with the negative control group. These findings demonstrate that MEFAS is safer than artesunate + mefloquine after acute administration in mice.

Testicular features of semi-confined Nellore bullssubjected to immunocastration / Características testiculares de touros Nelore semiconfinadossubmetidos a imunocastração

This study aimed to evaluate the effect of immunocastration on the scrotal circumference, as well as in the macroscopic and microscopic testicular features in Nellore bulls bred under a semi-confining system for 91 days. Sixty animals were divided into two groups: 30 intact animals (non-vaccinated) and 30 immunocastrated animals. The immunocastrated animals were treated with two anti-GnRH vaccine doses (BoprivaTM - Zoetis Australia Pty Ltd). The scrotal circumference (SC) was measured on days 0 and 56. Testicular parameters: biometry traits (length, width, and height ­ cm; volume ­ cm3) and weight (g) was measured on day 91. Three testicles from each group were histologically processed to find the seminiferous tubule diameter and epithelium thickness (µm). Data were analyzed in the Minitab® 19 statistical software. The macroscopic features (SC, and testes weight, volume, length, width, and height) and the microscopic features (seminiferous tubule diameter and epithelium thickness) were compared between the two groups through Student's t-test and Mann-Whitney test. All the values were lower (p<0.05) in animals immunized against GnRH, except for the right testis width and seminiferous tubule diameter. Results indicate that immunization against GnRH affected testicular development.(AU)

O objetivo do presente estudo foi avaliar o efeito da imunocastração no perímetro escrotal, assim como nas características testiculares macroscópicas e microscópicas de touros Nelore criados em um sistema de semiconfinamento por 91 dias. Sessenta animais foram divididos em dois grupos: 30 animais inteiros (não vacinados) e 30 imunocastrados. Os animais imunocastrados receberam duas doses de uma vacina anti-GnRH (Bopriva® - Zoetis Ltda). O perímetro escrotal (PE) foi mensurado nos dias 0 e 56. Parâmetros testiculares: atributos biométricos (comprimento, largura e altura ­ cm; volume cm3) e peso (g) foram medidos no dia 91. Três testículos de cada grupo foram processados para histologia clássica a fim de avaliar o diâmetro e a espessura do epitélio dos túbulos seminíferos (µm). Os dados foram analisados pelo software estatístico Minitab® 19. As médias das características macroscópicas (PE, peso, volume, comprimento, largura e altura dos testículos) e das microscópicas (diâmetro e espessura do epitélio dos túbulos seminíferos) foram comparadas entre os dois grupos pelos testes t de Student e Mann-Whitney. Todos os valores foram menores (p<0,05) em animais imunizados contra o GnRH, exceto a altura média do testículo direito e o diâmetro do túbulo seminífero. Os resultados indicam que a imunização contra o GnRH afeta o desenvolvimento testicular.(AU)

Heart-Protective Effects of Echinodorus grandiflorus in Rabbits That Are Fed a High-cholesterol Diet.

Excess weight and dyslipidemia are among the most serious health problems in Western societies. These conditions enhance the risk of cardiac disease and have been linked with a higher prevalence of cardiac arrhythmias and sudden death. The present study investigated the cardioprotective effects of Echinodorus grandiflorus on ventricular remodeling in rabbits that were fed a 1% cholesterol-rich diet. We first obtained an ethanol-soluble fraction of E. grandiflorus and performed a detailed phytochemical study by liquid chromatography-DAD/ESI-MS. For 60 days, male rabbits were fed the cholesterol-rich diet or a diet without the addition of cholesterol. After 30 days, different groups of rabbits were treated with the ethanol-soluble fraction of E. grandiflorus (10, 30, and 100 mg/kg, p. o.), simvastatin (2.5 mg/kg), or vehicle once daily for 30 days. At the end of 60 days, the serum lipoprotein ratio, electrocardiographic profile, histopathological alterations, and the cardiac antioxidant defense system were investigated. Echocardiographic analysis showed morphological and functional alterations in cholesterol-rich diet-fed animals, indicating left ventricle hypertrophy. The total cholesterol/high-density lipoprotein ratio and low-density lipoprotein/high-density lipoprotein ratio were significantly higher in cholesterol-rich diet-fed rabbits. Myocardial flaccidity, fatty degeneration, and concentric left ventricular hypertrophy were observed. An increase in lipid peroxidation levels, a decrease in superoxide dismutase activity, and a decrease in reduced glutathione levels were observed in the myocardium of all cholesterol-rich diet-fed rabbits. Treatment with the ethanol-soluble fraction of E. grandiflorus, especially the highest dose, significantly reduced all of these alterations, thus demonstrating the cardioprotective effect of the ethanol-soluble fraction of E. grandiflorus on cardiac changes that are induced by a cholesterol-rich diet.

Atheroprotective effects of Cuphea carthagenensis (Jacq.) J. F. Macbr. in New Zealand rabbits fed with cholesterol-rich diet.

ETHNOPHARMACOLOGICAL RELEVANCE: Although Cuphea carthagenensis (Jacq.) J. F. Macbr. is used in Brazilian folk medicine in the treatment of atherosclerosis and circulatory disorders, no study evaluating these effects has been conducted. The aim of this study was to evaluate the possible hypolipemiant and antiatherogenic activity of the ethanol soluble fraction obtained from C. carthagenensis (ES-CC) in an experimental atherosclerosis model using New Zealand (NZ) rabbits undergoing cholesterol-rich diet (CRD). MATERIAL AND METHODS: Dyslipidemia and atherogenesis were induced by administration of standard commercial diet increased of 1% cholesterol (CRD) for 8 weeks. ES-CC was orally administered at doses of 10, 30 and 100mg/kg, once daily for four weeks, starting from the 4th week of CRD diet. Body weight measurements were weekly carried out from the beginning of experiments for 8 weeks. Serum levels of triglyceride (TG), total cholesterol (TC) and their fractions (LDL-C, VLDL-C and HDL-C) were measured at the beginning of experiments and at weeks four and eight. After euthanasia of rabbits, aorta segments (aortic arc, thoracic, abdominal and iliac segments) were macroscopically and microscopically evaluated and the intima and media layers of the arteries were measured. Additionally, the antioxidant activity of ES-CC and its influence on the functioning of hepatic antioxidant enzymes were also determined. RESULTS: CRD induced dyslipidemia and major structural changes in the aortic wall. In addition, an increase in lipid peroxidation and a reduction of hepatic glutathione and serum nitrite levels were observed. Treatment with ES-CC was able to prevent the increase in TC, LDL-C, VLDL-C levels and triglycerides and promoted an increase in HDL-C levels in NZ rabbits. These effects were accompanied by a significant reduction in oxidative stress and modulation of the catalase and superoxide dismutase function. Moreover, the intima and media layers of the arterial segments were significantly reduced by ES-CC treatment. CONCLUSIONS: This study demonstrated that ES-CC reduces serum lipids and hepatic oxidative stress when orally administered to NZ rabbits. In addition, it was able to prevent the development of CRD-induced atherosclerosis.