Antimicrobial and toxicological evaluation of Origanum vulgare: an in vivo study / Avaliação antimicrobiana e toxicológica de Origanum vulgare: um estudo in vivo

Abstract Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin alongwith marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases.

Resumo Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

Bioactive phenolic compounds and organic acids in the decoction of fruits and leaves of Schinus areira L / Compuestos fenólicos bioactivos y ácidos orgánicos en la decocción de frutos y hojas de Schinus areira L

Leaf and fruit decoctions of Schinus areira L. from northwest Argentina were investigated here. Phenolic compounds and organic acids were analyzed by HPLC. Antioxidant capacity and α-glucosidase inhibition were determined by using in vitro tests. The general toxicity was assessed against Artemia salina nauplii. Hyperoside and 3 O-caffeoylquinic acid in leaf decoctions; gallic acid and catechin in fruit decoction were the major phenolic compounds. Malic and citric acids were the main organic acid quantified in the leaf and fruit decoctions, respectively. Fruit decoction had a relatively important content of shikimic acid, precursor of Tamiflu. Leaf decoction presents a greater richness in bioactive compounds with antiradical activity against DPPH●, O2●-and ●NO radicals. S. areira leaves and fruits had α-glucosidase inhibitory activity comparable to hyperoside and acarbose. Fruit decoction was not eco-toxic; leaf decoction showed significant eco-toxic activity and could be chosen for the search of other bioactive compounds with pharmacological activity.

Se investigaron decocciones de hojas y frutos de Schinus areira L. del noroeste de Argentina. Compuestos fenólicos y ácidos orgánicos se analizaron mediante HPLC. Capacidad antioxidante e inhibición de α-glucosidasa se determinaron in vitro. Se evaluó toxicidad general con Artemia salina. Los principales compuestos fenólicos fueron hiperósido y ácido 3 O-cafeoilquínico en hojas y ácido gálico y catequina en frutos. Los principales ácidos orgánicos cuantificados fueron málico en hojas y cítrico en frutos. Ácido shikímico, precursor del Tamiflu está presente en decocción de frutos con un contenido relativamente importante. La de hojas presenta una mayor riqueza en compuestos bioactivos con actividad antirradicalaria frente a DPPH●, O2●-y ●NO. Las hojas y frutos de S. areira tenían una actividad inhibidora de la α-glucosidasa comparable a la de hiperósido y acarbosa. La decocción de frutas no fue eco-tóxica, pero sí la de hojas que podría ser fuente de compuestos bioactivos con actividad farmacológica.

Extracts and purified substances of Cabralea canjerana inhibit hatching and extracts of Schinus terebinthifolius kill juveniles of meloidogyne incognita

Nematicidal substances have been identified from plants and are potentially useful for the management of plant-parasitic nematodes. Cabralea canjerana, (Meliaceae) and Schinus terebinthifolius (Anacardiaceae) produce bioactive compounds during their secondary metabolism and little is known about the effect of such substances on plant-parasitic nematodes. In the present study, we assessed the effect of aqueous and ethanolic extracts of C. canjerana and S. terebinthifolius at 1% (m:v) and purified substances from C. canjerana (gedunin, ocotillone, cabraleadiol, a mixture of ocotillone + cabraleadiol and a mixture of shoreic acid + eichlerianic acid) on hatching and mortality of Meloidogyne incognita juveniles. Aqueous extracts of C. canjerana fruits and seeds reduced hatching by 70.3 to 95.7%. Aqueous extracts of S. terebinthifolius fruits killed 42.8 to 77.1% of juveniles. The purified substances of C. canjerana inhibited the hatching of M. incognita from 57 to 90% and did not increase the mortality of juveniles. Therefore, C. canjerana extracts and its purified substances reduce M. incognita hatching and aqueous extracts of S. terebinthifolius kill J2 of this nematode.

Phytotoxicity of plant extracts of Vismia japurensis cultivated in vivo and in vitro / Fitotoxicidade de extratos de plantas de Vismia japurensis cultivadas in vivo e in vitro

Plants that produce secondary metabolites with allelopathic activity or phytotoxicity can be biotechnologically important, serving as sources of allelochemicals, and thus contributing to the agroindustrial sector. Vismia japurensis (Hypericaceae) is an Amazonian species that grows in clumps called vismiais, from which most other plants are absent. Accordingly, the objective of this study was to identify possible phytotoxicity effects of hexane and methanol extracts of Vismia japurensis leaves and branches in vivo and from seedlings grown in vitro on Lactuca sativa. In addition, fresh and dry leaves were assayed by the sandwich method in order to determine their ability to release allelochemicals. The hexanic extract from in vitro seedlings reduced germination by 10%, while the methanol extract produced a 16% reduction in germination speed. Root growth of Lactuca sativa was inhibited by 64.7% when subjected to hexane leaf extract, by 39.3% under the influence of hexane branch extract, and by 96.09% for in vitro seedling hexanic extract. When analysed by thin layer chromatography and 1H nuclear magnetic resonance, extracts showed evidence of terpenes, anthraquinones and flavonoids, with greater intensity of signals in the aromatic region of in vitro seedling hexanic extract. Clearly, Vismia japurensis has a high biotechnological potential in terms of the production of substances of low polarity with capacity to interfere in plant development.

Plantas que produzem metabólitos secundários com atividade alelopática ou fitotóxica podem ser biotecnologicamente importantes, servindo como fontes de aleloquímicos e, assim, contribuindo para o setor agroindustrial. Vismia japurensis (Hypericaceae) é uma espécie amazônica que cresce em grupos, formando vismiais. Assim, o objetivo deste estudo foi identificar possíveis efeitos fitotóxicos de extratos hexânicos e metanólicos de folhas e ramos de Vismia japurensis in vivo e de plântulas cultivadas in vitro sobre Lactuca sativa. Além disso, folhas frescas e secas foram analisadas pelo método sanduíche, a fim de determinar sua capacidade de liberação de aleloquímicos. O extrato hexânico de plântulas in vitro reduziu a germinação em 10% e o extrato metanólico promoveu uma redução de 16% na velocidade de germinação. O crescimento radicular de Lactuca sativa foi inibido em 64,7% quando submetido ao extrato hexânico das folhas, em 39,3% sob influência do extrato hexânico dos galhos e em 96,09% para o extrato de hexânico das plântulas in vitro. Quando analisados por cromatografia em camada delgada e ressonância magnética nuclear de 1H, os extratos mostraram evidências de terpenos, antraquinonas e flavonoides, com maior intensidade de sinais na região aromática do extrato hexânico das plântulas in vitro. Assim, Vismia japurensis possui elevado potencial biotecnológico em termos de produção de substâncias de baixa polaridade com capacidade de interferência no desenvolvimento de plantas.

Cytotoxic effect of Vernonanthura polyanthes leaves aqueous extracts / Efeito citotóxico do extrato aquoso de folhas de Vernonanthura polyanthes

Abstract Vernonanthura polyanthes, popularly known as assa-peixe, is a medicinal plant that has been widely used by Brazilian Cerrado population for treatment of diseases without a detailed evaluation of their effectiveness, toxicity, and proper dosage. Thus, more studies investigating the safety of V. polyanthes aqueous extract before the use are needed. The purpose of this study was to evaluate the toxicity, cytotoxicity and genotoxicity of V. polyanthes leaves aqueous extract using the Artemia salina and Allium cepa assays. For the A. salina assay, three groups of 10 larvae were exposed to V. polyanthes leaves aqueous extract at the concentrations of 5, 10, 20, 40, and 80 mg/ml. For the A. cepa assay, 5 onion bulbs were exposed to V. polyanthes leaves aqueous extract at 10, 20, and 40 mg/ml, and then submitted to macroscopic and microscopic analysis. As result it was identified a toxicity and cytotoxicity of V. polyanthes dependent on the extract concentration. The A. salina assay suggests that the concentration of 24 mg/ml of the V. polyanthes extract is able to kill 50% of naupllis; while the A. cepa assay suggests that V. polyanthes leaves aqueous extract is toxic at concentrations higher than 20 mg/ml; however the cytotoxic effect in A. cepa root cells was observed at 40 mg/ml of the extract. It is important to say that the V. polyanthes leaves aqueous extract concentration commonly used in popular medicine is 20 mg/ml. Thus, the popular concentration used is very close to toxicity limit in A. salina model (24 mg/ml) and is the concentration which showed toxic effect in A. cepa root cells (20 mg/ml). No genotoxic activity of V. polyantes leaves aqueous extract was observed in the conditions used in this study. Because of the antiproliferative action and no genotoxic activity, V. polyanthes leaves aqueous extract may present compounds with potential use for human medicine. However more detailed studies need to be performed to confirm this potential.

Resumo Vernonanthura polyanthes, popularmente conhecida como assa-peixe, é uma planta medicinal amplamente utilizada pela população brasileira do Cerrado para o tratamento doenças, sem uma avaliação detalhada de sua eficácia, toxicidade e dosagem adequada. Dessa forma, são necessários estudos para investigar a segurança do uso do extrato aquoso de V. polyanthes. O objetivo deste estudo foi avaliar a toxicidade, citotoxicidade e genotoxicidade do extrato aquoso de folhas de V. polyanthes utilizando os ensaios de Artemia salina e Allium cepa. Para o ensaio de A. salina, três grupos de 10 larvas foram expostos ao extrato aquoso de folhas de V. polyanthes nas concentrações de 5, 10, 20, 40 e 80 mg/ml. Para o ensaio de A. cepa, 5 bulbos de cebola foram expostas ao extrato aquoso de folhas de V. polyanthes nas concentrações de 10, 20 e 40 mg/ml, e então submetidos a análise macroscópica e microscópica. O ensaio de A. salina sugere que a concentração de 24 mg/ml do extrato de V. polyanthes é capaz de matar 50% dos náuplios; enquanto o ensaio de A. cepa sugere que o extrato aquoso das folhas de V. polyanthes é tóxico em concentrações superiores a 20 mg/ml. O efeito citotóxico nas células da raiz de A. cepa foi observado apenas na concentração de 40 mg/ml. É importante dizer que a concentração de extrato aquoso de folhas de V. polyanthes comumente usada na medicina popular é de 20 mg/ml. Assim, a concentração popular utilizada está muito próxima do limite de toxicidade no modelo de A. salina (24 mg/ml) e é a mesma concentração que apresentou efeito tóxico nas células da raiz de A. cepa (20 mg/ml). Não foi observada atividade genotóxica do extrato aquoso de folhas de V. polyantes nas condições utilizadas neste trabalho. Por causa da ação antiproliferativa e ausência de atividade genotóxica, o extrato aquoso de folhas de V. polyanthes pode ser uma boa fonte natural de compostos antitumorais e pode apresentar potencial para uso na medicina. No entanto, estudos mais detalhados precisam ser realizados para confirmar esse potencial.

Phytochemical composition, antifungal activity, in vitro and in vivo toxicity of Syzygium cumini (L) Skeels leaves extract / Composición fitoquímica, actividad antifúngica, toxicidad in vitro e in vivo del extracto de hojas de Syzygium cumini (L) Skeels

This study determined phytochemical composition, antifungal activity and toxicity in vitro and in vivo of Syzygium cumini leaves extract (Sc). Thus, was characterized by gas chromatography coupled to mass spectrometry and submitted to determination of Minimum Inhibitory (MIC) and Fungicidal concentrations (MFC) on reference and clinical strains of Candida spp. and by growth kinetics assays. Toxicity was verified using in vitro assays of hemolysis, osmotic fragility, oxidant and antioxidant activity in human erythrocytes and by in vivo acute systemic toxicity in Galleria mellonella larvae. Fourteen different compounds were identified in Sc, which showed antifungal activity (MIC between 31.25-125µg/mL) with fungistatic effect on Candida. At antifungal concentrations, it demonstrated low cytotoxicity, antioxidant activity and neglible in vivotoxicity. Thus, Sc demonstrated a promising antifungal potential, with low toxicity, indicating that this extract can be a safe and effective alternative antifungal agent.

Este estudio determinó la composición fitoquímica, la actividad antifúngica y la toxicidad in vitro e in vivo del extracto de hojas de Syzygium cumini (Sc). Así, se caracterizó mediante cromatografía de gases acoplada a espectrometría de masas y se sometió a determinación de Concentraciones Mínimas Inhibitorias (CMI) y Fungicidas (MFC) sobre cepas de referencia y clínicas de Candida spp. y mediante ensayos de cinética de crecimiento. La toxicidad se verificó mediante ensayos in vitro de hemólisis, fragilidad osmótica, actividad oxidante y antioxidante en eritrocitos humanos y por toxicidad sistémica aguda in vivo en larvas de Galleria mellonella. Se identificaron catorce compuestos diferentes en Sc, que mostraron actividad antifúngica (CMI entre 31.25-125 µg/mL) con efecto fungistático sobre Candida. En concentraciones antifúngicas, demostró baja citotoxicidad, actividad antioxidante y toxicidad in vivo insignificante. Por lo tanto, Sc demostró un potencial antifúngico prometedor, con baja toxicidad, lo que indica que este extracto puede ser un agente antifúngico alternativo seguro y eficaz.

Inhibition of α-glucosidase, pancreatic lipase, and antioxidant property of Myrcia hatschbachii D. Legrand containing gallic and ellagic acids / Inhibición de la α-glucosidasa, la lipasa pancreática y la propiedad antioxidante de Myrcia hatschbachii D. Legrand que contiene ácidos gálico y elágico

Several species of the Myrcia genus have been used in folk medicine to treat diabetes. Therefore, the aim of this work was to investigate the inhibitory activity of α-glucosidase and pancreatic lipase in the crude extract (EBF) and in the ethyl acetate fraction (FFA) of Myrcia hatschbachii, as well as to identify isolated phenolic compounds and to evaluate the antioxidant property and preliminary in vitro toxicity against Artemia salina. EBF (IC50: 3.21 µg/mL) and FFA (IC50: 1.14 µg/mL) showed inhibitory activity superior to acarbose (IC50: 193.65 µg/mL). In addition, they showed inhibitory effects of pancreatic lipase (IC50: 556.58 µg/mL for EBF and 532.68 µg/mL for FFA), antioxidant potential, absence of preliminary toxicity and presence of gallic andellagic acids in FFA. The relevant results in the inhibition of α-glucosidase and pancreatic lipase motivate new studies for the development of herbal medicines that assist in the treatment of diabetic patients.

Varias especies del género Myrcia se han utilizado en la medicina popular para tratar la diabetes. Por lo tanto, el objetivo de este trabajo fue investigar la actividad inhibitoria de la α-glucosidasa y la lipasa pancreática en el extracto crudo (EBF) y en la fracción de acetato de etilo (FFA) de Myrcia hatschbachii, así como identificar compuestos fenólicos aislados y evaluar la propiedad antioxidante y toxicidad in vitro preliminar contra Artemia salina. EBF (IC50: 3.21 µg/mL) y FFA (IC50: 1.14 µg/mL) mostraron una actividad inhibitoria superior a la acarbosa (IC50: 193.65 µg/mL). Además, mostraron efectos inhibitorios de la lipasa pancreática (IC50: 556.58 µg/mL para EBF y 532.68 µg/mL para FFA), potencial antioxidante, ausencia de toxicidad preliminar y presencia de ácidos gálico y elágico en FFA. Los resultados relevantes en la inhibición de la α-glucosidasa y la lipasa pancreática motivan nuevos estudios para el desarrollo de medicamentos a base de hierbas que ayudan en el tratamiento de pacientes diabéticos.

Toxicity evaluation of water extract of tissue-cultured Taraxacum formosanum by acute, subacute administration, and Ames test

BACKGROUND: Taraxacum species (commonly known as dandelion) used as herbal medicine have been reported to exhibit an antiproliferative effect on hepatoma cells and antitumor activity in non-small-cell lung cancer cells. Although several investigations have demonstrated the safety of Taraxacum officinale, the safety of tissue-cultured plants of T. formosanum has not been assessed so far. Therefore, the present study examines the safety of the water extract of the entire plant of tissue cultured T. formosanum based on acute and subacute toxicity tests in rats, as well as the Ames tests. RESULTS: No death or toxicity symptoms were observed in the acute and subacute tests. The results of the acute test revealed that the LD50 (50% of lethal dose) value of the T. formosanum water extract for rats exceeded 5 g/kg bw. No abnormal changes in the body weight, weekly food consumption, organ weight, or hematological, biochemical, and morphological parameters were observed in the subacute toxicity test. Thus, the no observed adverse effect level (NOAEL) of T. formosanum water extract was estimated to be higher than 2.0 g/kg. Finally, the results of the Ames test revealed that T. formosanum water extract was not genotoxic at any tested concentration to any of five Salmonella strains. CONCLUSIONS: The water extract of tissue-cultured T. formosanum was non-toxic to rats in acute and subacute tests and exhibited no genotoxicity to five Salmonella strains.

Use of erythrocytes in cytotoxicity and toxicity assays of medicinal plant extracts: analysis of their application and bibliometric study / Utilización de eritrocitos en ensayos de citotoxicidad y toxidez de extractos de plantas medicinales: análisis de su aplicación y estudio bibliométrico

Plant species have been used for therapeutic purposes since ancient times and are still in use today since these products represent a source of raw material for the production of phytotherapeutic formulations. Screening and investigation of plants with pharmacological potential require the evaluation of characteristics related to their action, efficacy and safety in different steps. Among these steps, pre- clinical trials are used to evaluate the properties of the test product in in vitro experiments, such as cytotoxicity assays. Within this context, this study consists of a bibliometric analysis of some in vitro cytotoxicity and toxicity assays in erythrocytes used during bioprospecting of medicinal plants. The results demonstrated the wide application of erythrocytes to evaluate the biological effects of medicinal plant extracts. The methods were found to be valid and effective for the preliminary investigation of the in vitro cytotoxicity and toxicity of plant products.

El uso de especies vegetales para fines terapéuticos es una práctica histórica y todavía bastante actual, ya que estos productos pueden representar una fuente de materia prima para la producción de formulaciones fitoterápicas. En investigación de plantas con potencial farmacológico requiere la evaluación de su acción, eficacia y seguridad, a través de diferentes etapas. Entre estas, en los ensayos preclínicos se evalúan las propiedades del producto-prueba en experimentos in vitro, tales como ensayos de citotoxicidad, entre otros. En este aspecto, el presente estudio consiste en un análisis bibliométrico acerca de algunas pruebas de citotoxicidad y toxicidad in vitro en eritrocitos realizados en los ensayos de bioprospección de plantas medicinales. Los resultados evidencian la amplia utilización de eritrocitos para la evaluación de los efectos biológicos de extractos de plantas medicinales, apuntándolos como métodos válidos y eficaces para la investigación preliminar de la citotoxicidad y toxicidad in vitro de productos vegetales.

Mechanisms of Toxicity and Cardiotoxicity of Alcohol Extract from Root, Stem and Leaf of Chloranthus Serratus / 法医学杂志

Objective To compare the differences of cardiotoxicity of alcohol extract from root, stem and leaf of Chloranthus serratus in the rats, and discuss preliminarily its mechanism of toxicity. Methods Rats were randomly divided into four groups: blank, root alcohol, stem alcohol and leaf alcohol, with 8 in each group. After 14 days of continuous intragastric administration, the body mass change curves were drawn. The cardiac coefficient was calculated. The contents of creatine kinase （CK）, creatine kinase isoenzyme （CK-MB）, lactate dehydrogenase （LDH） and α-hydroxybutyrate dehydrogenase （α-HBDH） as well as the content changes of oxidative stress indexes - total superoxide dismutase （T-SOD） and malondialdehyde （MDA） in the serum of rats were detected. The cardiac pathomorphology changes in the rats were observed. The expression of intercellular adhesion molecule （ICAM-1） and heme oxygenase （HO-1） in myocardial tissue was detected. Results Body mass growth rate： stem alcohol group was the smallest, followed by leaf alcohol group. The difference of cardiac coefficient of every group had no statistical significance （P>0.05）. The myocardial tissues of stem alcohol group suffered the most serious damage, followed by the leaf alcohol group. The contents of CK, CK-MB, LDH and α-HBDH in stem alcohol group increased （P<0.05）. The increase of MDA content and decrease of T-SOD content in stem alcohol group had statistical significance compared with the blank group and root alcohol group, while the leaf alcohol group only had statistical significance in the decrease of T-SOD content compared with the blank group （P<0.05）. The positive expression of ICAM-1 enhanced and the expression of HO-1 protein decreased in every group after the intervention of different extracts. The change trend was stem alcohol > leaf alcohol > root alcohol group. Conclusion The alcohol extract from the stem has the highest cardiotoxicity, followed by the leaf extract, and its mechanism of toxicity may be related to oxidative stress.

Efecto analgésico, caracterización fitoquímica y análisis toxicológico del extracto etanólico de hojas de pereskia lychnidiflora / Analgesic effect, phytochemical characterization and toxicological analysis of ethanolic extract of pereskia lychnidiflora leaves

RESUMEN Objetivo Evaluar el efecto analgésico del extracto etanólico de las hojas de Pereskia lychnidiflora, la prospección de metabolitos secundarios y el análisis toxicológico. Materiales y métodos La actividad analgésica fue evaluada mediante la prueba del ácido acético y la formalina en ratones NIH a una concentración de 30, 50 y 100 mg/kg de peso corporal, utilizando como control Ibuprofeno a 200 mg/kg y agua destilada como blanco. La prospección de metabolitos secundarios se realizó por el método de cromatografía de capa fina y la toxicidad del extracto fue evaluada in vivo según la dosis máxima de 2000 mg/kg de peso corporal. Resultados La prospección fitoquímica determinó la presencia de alcaloides, taninos, triterpenos y esteroles como mayores constituyentes químicos. Se determinó que el extracto etanólico de Pereskia lychnidiflora posee una actividad analgésica similar al Ibuprofeno. No se observaron signos de toxicidad en los ratones de experimentación y se clasifica el extracto como no tóxico con una DL50 mayor de 2000 mg/kg. Conclusión El extracto etanólico de Pereskia lychnidiflora tiene un efecto analgésico antiinflamatorio que podría estar condicionado por la presencia de alcaloides, taninos y esteroles (terpenoides) presentes en esta especie vegetal y puede ser clasificado como no tóxico.

ABSTRACT Objective To evaluate the analgesic effect of the ethanolic extract of the leaves of Pereskia lychnidiflora, the prospection of secondary metabolites and the toxicologic analysis. Materials and Methods Analgesic activity was evaluated by testing acetic acid and formalin in NIH mice at a concentration of 30, 50 and 100 mg/kg body weight, using Ibuprofen control at 200 mg/kg and distilled water as the target. Secondary metabolites were prospected using the thin layer chromatography method and the toxicity of the extract was evaluated in vivo according to the maximum dose of 2,000 mg/kg body weight. Results Phytochemical prospecting determined the presence of alkaloids, tannins, triterpenes, and sterols as major chemical constituents. The ethanolic extract of Pereskia lychnidiflora was found to have an analgesic activity similar to ibuprofen. No signs of toxicity were observed in the experimental mice and the extract is classified as non-toxic with a DL50 greater than 2,000 mg/kg. Conclusions The ethanolic extract of Pereskia lychnidiflora has an anti- inflammatory analgesic effect that could be conditioned by the presence of alkaloids, tannins, and sterols (terpenoids) present in this species and can be classified as non-toxic.

Effects of Leucaena leucocephala (lamk. ) shoot tips plus young leaf extract containing mimosine on reproductive system of male rats / Efectos de los brotes de Leucaena leucocephala (lamk. ) y del extracto de hojas jóvenes que contiene mimosina en el sistema reproductivo de ratas macho

This study aimed to determine the mimosine level and examine the male reproductive toxicity effects of Leucaena leucocephala (LL) shoot tips plus young leaf extract. Mimosine level in LL extract was determined by thin layer chromatography before administration in animals. Male rats were divided into control and LL (1,500 mg/KgBW) groups (n = 6). After 60 days of experiment, serum sex hormones, sperm quality, and testicular histopathology were assayed and observed. Malondialdehyde (MDA) level and expressions of steroidogenic acute regulatory (StAR) and phosphorylated proteins in testicular lysate were examined by western blotting. Results showed that mimosine levels in LL extract was 17.35 ± 1.12 % of dry weight. LL significantly decreased FSH & LH levels, sperm qualities, and seminiferous tubule diameter compared to the control (p<0.05). Seminiferous tubular atrophies, germ cell sloughing, and degenerations were observed in LL group. In addition, testicular MDA level and StAR protein expression were significantly decreased in LL group. LL extract could increase the expression of a 50 kDa phohorylated protein in testicular lysate. In conclusion, LL extract has mimosine and reproductive toxicity effects on males.

Este trabajo tuvo como objetivo determinar el nivel de mimosina y examinar los efectos de la toxicidad reproductiva de los brotes de Leucaena leucocephala (LL), más el extracto de hojas jóvenes, en ratas macho. El nivel de mimosina en el extracto de LL se determinó mediante cromatografía en capa fina antes de la administración en animales. Las ratas se dividieron en grupos de control y LL (1,500 mg / kgBW) (n = 6). Después de 60 días, se analizaron y observaron las hormonas sexuales séricas, la calidad de los espermatozoides y la histopatología testicular. A través de Western Blot se examinaron el nivel de malondialdehído (MDA), las expresiones de reguladores agudos esteroidogénicos (StAR) y las proteínas fosforiladas en el lisado testicular. Los resultados mostraron que los niveles de mimosina en el extracto de LL fueron 17.35 ± 1.12 % del peso seco. LL disminuyó significativamente los niveles de FSH y LH, la calidad de los espermatozoides y el diámetro de los túbulos seminíferos en comparación con el control (p <0,05). Se observaron atrofias en los túbulos seminíferos, desprendimiento de células germinales y degeneraciones en el grupo LL. Además, el nivel de MDA testicular y la expresión de la proteína StAR se redujeron significativamente en el grupo LL. El extracto de LL podría aumentar la expresión de la proteína fosforilada de 50 kDa en el lisado testicular. En conclusión, el extracto de LL tiene mimosina y efectos de toxicidad reproductiva en los hombres.

Catha edulis forsk mediates embryotoxic effects in rats: an experimental study / Catha edulis forsk modera los efectos embriotóxicos en ratas: un estudio experimental

Catha edulis Forsk leaves (Khat) is a flowering plant. A high proportion of the adult population in the Arabian Peninsula and the Horn of Africa chews it for its mild stimulant effect. The aim of the current study was to investigate the embryotoxic and teratogenic effects of the Khat extract using 60 female pregnant rats. These were divided to a Khat extract-treated group and a control group. Methanolic extract of Khat was orally given to the treated group 4 days before mating and up to day 16 of pregnancy with a dose of 100 mg/kg. Our results showed that significant number of embryos of the Khat-treated mothers were malformed and different in size and shape compared to embryos from the mothers of the control group. At day 8 of pregnancy, malformed embryos had ill developed primitive layers. By day 10 of pregnancy, neural tube and the somite were not formed compared to the control embryos. At later stages of pregnancy, embryos of the Khat-treated mothers appeared severely abnormal with opened neural groove and visceral pouches. Disrupted normal neural tube development, undifferentiated brain vesicles, incomplete closure of the brain flexures were also observed in these embryos. Highly significant increase in the number of the resorbed embryos of the Khat-treated mothers were observed (P < 0.01). The resorbed embryos appeared as a cellular collection in their placenta with some of their decidua had no visible embryonic tissues. In conclusions, Khat induced embryotoxic effects as well as severely affected the early normal embryonic development in rat.

Catha edulis (Khat) es una planta floreciente. Una alta proporción de la población adulta en la Península Arábiga y el Cuerno de África la mastica por su efecto estimulante. El objetivo del presente estudio fue investigar los efectos embriotóxicos y teratogénicos del extracto de Khat utilizando 60 ratas hembras preñadas. Estas se dividieron en un grupo tratado con extracto de Khat y un grupo control. El extracto metanólico de Khat se administró por vía oral al grupo tratado 4 días antes del apareamiento y hasta el día 16 de preñez con una dosis de 100 mg / kg. Los resultados mostraron que una cantidad significativa de embriones de las madres tratadas con Khat tenían malformaciones y eran diferentes en tamaño y forma en comparación con los embriones de las madres del grupo control. En el día 8 de preñez, los embriones malformados tenían capas primitivas mal desarrolladas. Para el día 10 de preñez, el tubo neural y el somito no se formaron en comparación con los embriones del grupo control. En etapas posteriores de la preñez, los embriones de las madres tratadas con Khat parecían severamente anormales con surcos neurales abiertos y bolsas viscerales. También se observaron alteraciones en el desarrollo normal del tubo neural, vesículas cerebrales indiferenciadas y el cierre incompleto de las flexiones cerebrales en estos embriones. Se observó un aumento altamente significativo en el número de embriones reabsorbidos de las madres tratadas con Khat (P <0,01). Los embriones reabsorbidos aparecieron como una colección celular en su placenta con algunas de sus deciduas sin tejidos embrionarios visibles. Khat indujo efectos embriotóxicos y afectó severamente el desarrollo embrionario normal temprano en la rata.

Avaliação citotóxica, genotóxica e mutagênica do extrato de Morinda citrifolia em diferentes concentrações sobre o teste Allium Cepa / Cytotoxic, genotoxic and mutagenic evaluation of Morinda citrifolia extract in different concentrations about the test Allium Cepa

Introdução: o uso de Morinda citrifolia (noni) realizado com várias finalidades, no entanto, sua eficácia ainda não é, plenamente, comprovada. Segundo a Agência Nacional de Vigilância Sanitária (2007), as publicações científicas sobre o suco de noni têm trazido muita controvérsia sobre sua segurança como alimento. Objetivos: o objetivo do presente trabalho foi avaliar quais concentrações de Morinda citrifolia não apresentam efeitos citotóxicos, genotóxicos e mutagênicos, possibilitando seu uso em futuras formas farmacêuticas. Metodologia: os frutos foram picados e desidratados em estufa. Em seguida o material foi pulverizado, obtendo-se o extrato seco. Foram utilizados bulbos de Alium cepa para testar as seguintes concentrações: controle negativo (água filtrada), 1 mg/mL (Tratamento 1), 1,5 mg/mL (Tratamento 2), 2 mg/mL (Tratamento 3), controle positivo (paracetamol 90 mg/mL). Resultados: os resultados encontrados na análise dos dados do extrato aquoso, demonstram que as três concentrações testadas de Morinda citrifolia apresenta atividade tóxica pela inibição do comprimento e pela diminuição do ciclo celular das raízes. Além disso, a Morinda citrifolia apresenta atividade citotóxica, devido à redução do índice mitótico, em todas as concentrações analisadas. Bem como, apresenta atividade genotóxica, nas duas maiores concentrações do extrato (1,5 mg/mL e 2,0 mg/mL). Conclusão: o presente estudo demonstrou que os extratos de Morinda citrifolia apresenta atividade citotóxica e genotóxica em todas as concentrações analisadas. É necessário realizar outros trabalhos para a avaliação da Morinda citrifolia em concentrações menores, para assim se estabelecer quais são as concentrações seguras de utilização do suco desse fruto

Mutagenicity and antimutagenicity of Salacia crassifolia (mart. Ex. Schult.) G. Don. evaluated by Ames test / Mutagenicitade e antimutagenicidade de Salacia crassifolia (mart. Ex. Schult.) G. Don. avaliadas pelo teste de Ames

Abstract Salacia crassifolia (Mart. Ex. Schult.) G. Don. is a bush which belongs to Celastraceae family and occurs specially in Brazilian Cerrado. Its leaves, stem, seeds and fruits are popularly used for several medicinal purposes, such as antitumoral, antirheumatic, anti-inflammatory and antimicrobial. In this study, the mutagenic and antimutagenic activities of S. crassifolia stem bark fractions (hexane, ethyl acetate and hydroalcoholic) were evaluated by the Ames mutagenicity assay in Salmonella typhimurium TA98 and TA100 strains. By the obtained results, all S. crassifolia fractions did not significantly increase the number of prototrophic revertants for histidine (His+) in both S. typhimurium strains tested (p > 0.05), suggesting absence of mutagenicity. Regarding antimutagenicity, the fractions ethyl acetate and hydroalcoholic significantly decreased the number of His+ revertants colonies induced by positive control for strain TA98 (p < 0.05), demonstrating protection against mutagenicity induced by 4-nitroquinolile1-oxide, whereas the hexane fraction did not show antimutagenic effect in this strain. In the TA100 strain, all fractions of S. crassifolia protected DNA against the harmful action of sodium azide, and the hexane fraction exhibited the greatest protection in this work. Thus, it's possible conclude that the fractions of S. crassifolia tested in this study could be used in chemoprevention.

Resumo Salacia crassifolia (Mart. Ex. Schult.) G. Don. é uma árvore que pertence à família Celastraceae e ocorre especialmente no Cerrado Brasileiro. Suas folhas, caule, sementes e frutos são popularmente utilizados para vários fins medicinais, tais como antitumoral, antirreumático, anti-inflamatório e antimicrobiano. Neste estudo, nós avaliamos as atividades mutagênica e antimutagênica de frações da casca do caule de S. crassifolia (hexânica, acetato de etila e hidroalcoólica) pelo ensaio de mutagenicidade de Ames em Salmonella typhimurium, cepas TA98 e TA100. Pelos resultados obtidos todas as frações de S. crassifolia não aumentaram significativamente o número de revertentes prototróficas para histidina (His+) em ambas as cepas de S. typhimurium testadas (p > 0.05), sugerindo ausência de mutagenicidade. Em relação à antimutagenicidade, as frações acetate de etila e hidroalcoólica reduziram significativamente o número de colônias revertentes His+ induzidas pelo controle positive para a cepa TA98 (p < 0.05), demonstrando sua ação protetora contra a mutagenicidade induzida por 4-nitroquinolile1-oxide, enquanto a fração hexânica não demonstrou efeito antimutagênico nesta cepa. Na cepa TA100, todas as frações de S. crassifolia protegeram o DNA contra a ação lesiva de azida sódica, e a fração hexânica exibiu a maior proteção desse trabalho. Assim, concluímos que as frações de S. crassifolia testadas neste estudo poderiam ser utilizadas em quimioprevenção.

Assessment of the antibacterial, cytotoxic and antioxidant activities of Morus nigra L. (Moraceae) / Avaliação das atividades antibacteriana, citotóxica e antioxidante de Morus nigra L. (Moraceae)

Abstract This study was carried out to assess the antibacterial, cytotoxic and antioxidant activities of extracts of Morus nigra L. HPLC was used to determine the fingerprint chromatogram of the crude ethanolic extract (Mn-EtOH). The antibacterial effect was assessed through the method of microdilution. The cytotoxicity was tested against human tumour cell lines using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The total phenolic and flavonoid contents were also assessed through the Folin-Ciocalteu and aluminum chloride methods, respectively. Antioxidant activities of the extracts were evaluated by using 2,2-diphenyl-1-picrylhydrazil (DPPH) radical scavenging and β-carotene-linoleic acid bleaching methods. The presence of phenolic compounds in Mn-EtOH was confirmed using HPLC. The extracts showed activity against most microorganisms tested. The extracts did not show any expressive antiproliferative effect in the assessment of cytotoxicity. The most significant total phenolic content was 153.00 ± 11.34 mg of gallic acid equivalent/g to the ethyl acetate extract (AcOEt). The total flavonoid content was 292.50 ± 70.34 mg of catechin equivalent/g to the AcOEt extract, which presented the best antioxidant activity (IC50 50.40 ± 1.16 μg/mL) for DPPH scavenging. We can conclude that this species shows strong antibacterial and antioxidant activities, as well as weak cytotoxic effects.

Resumo Este estudo foi realizado para avaliar as atividades antibacteriana, citotóxica e antioxidante de extratos de Morus nigra L. HPLC foi utilizado para determinar o perfil de compostos fenólicos do extrato etanólico bruto (Mn-EtOH). O efeito antibacteriano foi avaliado através do método de microdiluição. A citotoxicidade foi testada contra linhagens celulares de tumores humanos utilizando o ensaio do brometo de 3-(4,5-dimetil-2-tiazolil)-2,5-difenil-2H-tetrazólio (MTT). O conteúdo total de compostos fenólicos e flavonoides também foi avaliado por meio dos métodos de Folin-Ciocalteu e cloreto de alumínio, respectivamente. A atividade antioxidante dos extratos foi avaliada por meio do sequestro do radical livre 2,2-difenil-1-picrilhidrazil (DPPH) e co-oxidação do sistema β-caroteno-ácido linoleico. A presença de compostos fenólicos em Mn-EtOH foi confirmada utilizando HPLC. Os extratos mostraram atividade contra a maioria dos microrganismos testados. Os extratos não mostraram qualquer efeito antiproliferativo expressivo na avaliação da citotoxicidade. O conteúdo fenólico total mais significativo foi de 153,00 ± 11,34 mg de equivalente de ácido gálico/g para o extrato acetato de etila (AcOEt). O conteúdo de flavonoides totais foi de 292,50 ± 70,34 mg de equivalente de catequina/g para o extrato AcOEt, que apresentou a melhor atividade antioxidante (IC50 50,40 ± 1,16 mg/mL) para o sequestro do DPPH. Podemos concluir que esta espécie apresenta forte atividade antibacteriana e antioxidante, bem como fraca atividade citotóxica.

Evaluación de la toxicidad subcrónica de la variedad colombiana de Smallanthus sonchifolius (Yacón) en ratas hembra / Subchronic toxicity evaluation of colombian variety of Smallanthus sonchifolius (Yacón) in female rats

Smallanthus sonchifolius (Yacón) es una planta usada comúnmente por largos periodos de tiempo con el fin de ayudar en el control de la diabetes y otros desordenes metabólicos, por lo que con el propósito de evaluar la toxicidad subcrónica de la variedad colombiana de esta planta, fueron tratadas 30 ratas hembra de 8 semanas de edad dividas en 6 grupos. A cada uno de ellos se administró durante 28 días una de las siguientes dosis de infusión acuosa liofilizada (500, 250 y 125 mg/kg de peso), evaluando paralelamente grupos control (positivo y negativo) e incluyendo entre ellos grupos con y sin dieta hipercalórica. Para el seguimiento del perfil metabólico de los animales, se tomaron muestras de sangre periódicamente durante el ensayo y se evaluaron los niveles séricos de: glucemia, triglicéridos, colesterol total y HDL. Además, también se realizó el control del peso, así como estudios comportamentales que incluyeron el Test de Irwin y el Test Hipocrático. Al final de estudio (28 días), se realizó el análisis anatomopatológico e histológico comparativo con el fin de detectar posibles daños tisulares. Como resultado pudo observase que el liofilizado, si bien puede tener un efecto antihiperglucemiante, no modificó significativamente el perfil lipídico. Además, a pesar de que la administración se hizo durante 28 días, no se observaron cambios comportamentales que evidencien toxicidad, pero sí pudieron observarse cambios histológicos en el tejido cardiaco como hialinización, separación y redondeo de fibras.

Abstract. Smallanthus sonchifolius (Yacón) is a plant commonly used over long periods of time to help control diabetes and other metabolic disorders. To assess the sub-chronic toxicity of the Colombia variety of this plant, it was tested on 30 eight-week-old female rats, divided into six groups. For 28 days each group was administered with the following doses: three groups with lyophi­lized aqueous infusion (500 mg, 250 mg and 125 mg per kg of weight), two control groups (positive and negative) being assessed in parallel; this groups receiving hyper-caloric diet, and the last group was the general control or normal control. To monitor the animals' metabolic profile, blood samples were taken from time to time during the test period, and the serum levels of glycemia, triglycerides, total cholesterol and HDL were measured. Weight tracking was also carried out, as well as behavioral studies, including the Irwin Test and the Hippocratic Test. At the end of the study (28 days), comparative anatomo-pathological and histological analyses were performed to detect possible tissue damage. The results showed that, although the lyophilized infusion could have an antihyperglycemic effect, it did not significantly change the lipid profile. Moreover, though the infusion was administered during 28 days, it was found that it did not lead to any behavioral changes indicating toxicity, but did produce in heart tissue histological changes such as hyalinization, separation and rounding of fibers.

Autophagy protects against neural cell death induced by piperidine alkaloids present in Prosopis juliflora (Mesquite)

ABSTRACT Prosopis juliflora is a shrub that has been used to feed animals and humans. However, a synergistic action of piperidine alkaloids has been suggested to be responsible for neurotoxic damage observed in animals. We investigated the involvement of programmed cell death (PCD) and autophagy on the mechanism of cell death induced by a total extract (TAE) of alkaloids and fraction (F32) from P. juliflora leaves composed majoritary of juliprosopine in a model of neuron/glial cell co-culture. We saw that TAE (30 µg/mL) and F32 (7.5 µg/mL) induced reduction in ATP levels and changes in mitochondrial membrane potential at 12 h exposure. Moreover, TAE and F32 induced caspase-9 activation, nuclear condensation and neuronal death at 16 h exposure. After 4 h, they induced autophagy characterized by decreases of P62 protein level, increase of LC3II expression and increase in number of GFP-LC3 cells. Interestingly, we demonstrated that inhibition of autophagy by bafilomycin and vinblastine increased the cell death induced by TAE and autophagy induced by serum deprivation and rapamycin reduced cell death induced by F32 at 24 h. These results indicate that the mechanism neural cell death induced by these alkaloids involves PCD via caspase-9 activation and autophagy, which seems to be an important protective mechanism.

Evaluation of the toxicity and hypoglycemic effect of the aqueous extracts of Cnidoscolus quercifolius Pohl

Diabetes mellitus is one of the most common chronic degenerative diseases, and it is estimated to increase worldwide to around 415 million and to impact 642 million in 2040. Research shows that some plants are sources of bioactive compounds against diabetes. Thus, the objective of this work was to evaluate the oral toxicity and the hypoglycemic effect of the aqueous extract of the leaves of Cnidoscolus quercifolius Pohl. Diabetes was induced in Swiss mice with streptozotocin and the mice were treated with an aqueous extract of C. quercifolius leaves for a period of 30 days. Phytochemical analysis showed that the extract was rich in flavonoids, catechins and triterpenoid, which did not show any mortality and behavioral alterations in mice treated with 200, 1000, and 2000 mg/kg body weight of the extract for 14 days. Histopathological analysis of organs (kidney, pancreas, liver) from mice treated with the 2000 mg/kg extract revealed no architectural change. In the present study, we found a 29% reduction in glucose levels in animals receiving 200 mg/kg body weight. These results are very promising because they showed that C. quercifolius had a hypoglycemic effect and did not present oral toxicity, thus being a new source of compounds for the control of diabetes.

Evaluation of the antiplasmodial and leishmanicidal potential of Myrciaria dubia (Myrtaceae) extract

Abstract INTRODUCTION: Malaria and leishmaniasis are prevalent in tropical regions, which have environmental characteristics that are highly favorable to protozoa and vectors of these diseases; the transmission of these infections in sub-tropical regions, although recognized, represents only a small fraction of cases. Plants are constantly being used in the search for and acquisition of new drugs, and many compounds derived from them have been used to combat various diseases. In this study, we evaluated the action of the dichloromethanolic extract of Myrciaria dubia leaves against the protozoa Plasmodium falciparum, Leishmania amazonensis, Leishmania braziliensis, and Leishmania chagasi through bioassays. METHODS The extract from M. dubia was tested for its anti-P. falciparum activity in an anti-histidine-rich protein II immunosorbent assay. The antileishmanial assays were performed using the resazurin method, while cytotoxicity against human hepatoma (HepG2) strain was determined using the colorimetric MTT [3-(4, 5-dimethyl-2- thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide] method. RESULTS The M. dubia extract presented a half-maximal inhibitory concentration equal to 2.35 (1.05)μg/mL for P. falciparum, 190.73 (6.41) μg/mL for L. amazonensis, and greater than equal to 200µg/mL for L. chagasi and L. braziliensis strains. The cytotoxic concentration for 50% of the cells was above 500μg/mL for HepG2, indicating no toxicity and greater selectivity against parasites. CONCLUSIONS The results obtained indicate the presence of antiplasmodial and leishmanicidal bioactive compounds in the dichloromethanolic extracts of M. dubia leaves, and point towards future studies to elucidate the mechanism of action for each physiological effect.