Crude extract of the tropical tree Gallesia integrifolia (Phytolaccaceae) for the control of Aedes aegypti (Diptera: Culicidae) larvae / Extracto crudo del árbol tropical Gallesia integrifolia (Phytolaccaceae) para el control de larvas de Aedes aegypti (Diptera: Culicidae)

Introduction: Phytoinsecticides are alternatives to control insects in different stages, Gallesia integrifolia (Spreng.) Harms, Phytolacaceae family, popularly known as pau d'alho, garlic tree, and guararema in Brazil, is known due to its strong alliaceous odor because of the presence of sulfur molecules in the plant. This species presents biological activity and potential insecticide effect that is still unexploited. Objective: This study aimed to evaluate the biological activity of the ethanolic crude extract from G. integrifolia leaves, flowers, and fruits on the control of Aedes aegypti third-stage larvae and pupae. Methods: The botanical material was collected in city Umuarama, Paraná, Brazil at the coordinates (23º46'16" S & 53º19'38" WO), and altitude of 442 m, the fruits of G. integrifolia were collected in May and the leaves and flowers in December 2017. The crude extracts of G. integrifolia leaves, flowers, and fruits were prepared by dynamic maceration technique. The chemical composition of the extracts was determined by gas chromatography coupled to a mass spectrometry. The insecticidal activity of the crude extracts of G. integrifolia were carried out on larvae and pupae of A. aegypti in concentrations between 0.001 to 25 000 mg/mL, and afterwards the lethal concentrations that kill 50 % (LC50) and 99.9 % (LC99.9) were determined by probit analysis. Anticholinesterase activity was determined by bioautographic method at concentrations from 0.000095 to 50 mg/mL. Results: The yield of G. integrifolia crude extracts were 8.2, 9.1, and 17.3 % for flowers, fruits, and leaves, respectively. The chemical composition of G. integrifolia extracts was characterized by presence of fatty acid esters, phytosterols, vitamins, oxygenated diterpenes and organosulfur compounds. The flower extract presented the high amount of sulfur compounds (20.2 %) such as disulfide, bis (2-sulfhydryl ethyl) (11.9 %), 2,3,5-trithiahexane (6.2 %), 1,2,4-trithiolane (1.1 %), and 2,4-dithiapentane (1.1 %). Regarding the insecticidal activity, flower extract showed highly active with LC99.9 of 0.032 mg/mL and LC99.9 of 0.969 mg/mL on A. aegypti larvae and pupae, respectively, and the highest inhibition of acetylcholinesterase enzyme (0.00019 mg/mL) ex situ. The flower extract presented anticholinesterase and larvicide activity, respectively, 12.8 % and 35.6 % greater than the control temephos. Conclusions: This study opens new perspectives on the use of extracts from G. integrifolia as a bioinsecticide alternative for the control of A. aegypti larvae and pupae.

Introducción: Los fitoinsecticidas son alternativas para el control de insectos en diferentes etapas, Gallesia integrifolia (Spreng.) Harms, familia Phytolacaceae, conocida popularmente como pau d'alho, árbol de ajo y guararema en Brasil, es conocida por su fuerte olor aliáceo debido a la presencia de moléculas de azufre en la planta. Esta especie presenta actividad biológica y potencial efecto insecticida que aún no está explotado. Objetivo: El objetivo de este estudio fue evaluar la actividad biológica del extracto crudo etanólico de las hojas, flores y frutos de G. integrifolia en el control de las larvas y pupas de la tercera etapa de Aedes aegypti. Métodos: El material botánico fue recolectado en la ciudad de Umuarama, Paraná, Brasil (23º46'16" S & 53º19'38" W), a una altitud de 442 m y los frutos de G. integrifolia fueron recolectados en mayo de 2017 y las hojas y flores en diciembre del mismo año. Los extractos crudos de hojas, flores y frutos de G. integrifolia se prepararon por técnica de maceración dinámica. La composición química de los extractos se determinó por cromatografía de gases acoplada a espectrometría de masas. La actividad insecticida de los extractos crudos de G. integrifolia fue evaluada en larvas y pupas de A. aegypti en concentraciones entre 0.001 a 25 000 mg/mL, y las concentraciones letales que matan 50 % (CL50) y 99.9 % (CL99.9) de larvas fueron determinados por análisis de probit. La actividad anticolinesterasa fue evaluada por método bioautográfico en concentraciones de 0.000095 a 50 mg/mL. Resultados: El rendimiento de los extractos crudos de G. integrifolia fue de 8.2, 9.1 y 17.3 % para flores, frutos y hojas, respectivamente. La composición química de los extractos de G. integrifolia se caracterizó por la presencia de ésteres de ácidos grasos, fitosteroles, vitaminas, diterpenos oxigenados y compuestos organosulfurados. El extracto de las flores presentó alta cantidad de compuestos de azufre (20.2 %) como disulfuro, bis(2-sulfhidril etilo) (11.9 %), 2,3,5-tritiahexano (6.2 %), 1,2,4-tritiolano (1.1 %) y 2,4-ditiapentano (1.1 %). En relación con la actividad insecticida, el extracto de las flores mostró una gran actividad con CL99.9 de 0.032 mg/mL y CL99.9 de 0.969 mg/mL en larvas y pupas de A. aegypti, respectivamente, y la inhibición más alta de la enzima acetilcolinesterasa (0.00019 mg/mL) ex situ. El extracto de las flores presentó actividad anticolinesterasa y larvicida, 12.8 y 35.6 %, respectivamente, mayor que el control temephos. Conclusiones: Este estudio abre nuevas perspectivas sobre el uso de extractos de G. integrifolia como alternativa bioinsecticida para el control de larvas y pupas de A. aegypti.

Insecticidal activity of Gallesia integrifolia (Phytolaccaceae) essential oil / Actividad insecticida del aceite esencial de Gallesia integrifolia (Phytolaccaceae)

This study evaluated the insecticidal activity of Gallesia integrifolia essential oil from fruits, leaves, and flowers against Aedes aegyptilarvae and pupae. The essential oil was obtained by hydrodistillation and characterized by gas chromatography-mass spectrometry. Sulfur compounds represented 95 to 99% of the essential oil from fruits, leaves, and flowers. Essential oil major compounds were 2,8-dithianonane (52.6%) in fruits, 3,5-dithiahexanol-5,5-dioxide (38.9%) in leaves, and methionine ethyl ester (45.3%) in flowers. The essential oils showed high activity against larvae, and low for pupae withLC99.9of 5.87 and 1476.67µg/mL from fruits; 0.0096 and 348.33 µg/mL from leaves and 0.021and 342.84 µg/mL from flowers, respectively. The main compound with insecticide activity is probably n-ethyl-1,3-dithioisoindole, from isoindole organothiophosphate class, found in greater amount in flower and leaf essential oil. The great insecticide activity of G. integrifolia essential oil suggests that this product is a natural insecticide.

Este estudio evaluó la actividad insecticida del aceite esencial de frutos, hojas y flores de Gallesia integrifolia contra larvas y pupas de Aedes aegypti. El aceite esencial se obtuvo por hidrodestilación y se caracterizó por cromatografía de gases acoplada a espectrometría de masas. Los compuestos de azufre representaron del 95 al 99% del aceite esencial de frutas, hojas y flores. Los compuestos principales del aceite esencial fueron 2,8-ditianonano (52,6%) en frutas, 3,5-ditiahexanol-5,5-dióxido (38,9%) en hojas y éster etílico de metionina (45,3%) en flores. Los aceites esenciales mostraron alta actividad contra larvas y baja para pupas con LC99.9de 5.87 y 1476.67µg/mL de frutos; 0,0096 y 348,33 µg/mL de hojas y 0,021 y342,84 µg/mL de flores, respectivamente. El principal compuesto con actividad insecticida es probablemente el n-etil-1,3-ditioisoindol, de la clase de los organotiofosfatos de isoindol, que se encuentra en mayor cantidad en el aceite esencial de flores y hojas. La gran actividad insecticida del aceite esencial de G. integrifolia sugiere que este producto es un insecticida natural.

Efecto de Petiveria alliacea y drogas colinérgicas sobre la habituación / Effects of Petiveria alliacea and cholinergic drugs on the habituation cognitive behavior

Petiveria alliacea (PA) have anxiolytic, antidepressant and cognitive effects. In the present paper the effect of PA water infusion and cholinergic drugs on cognitive behavior were studied. For that, 40 male NMRI mice were divided in 4 groups: Control (n=10), Drug Control (n=10), PA (n=10) and PA plus Drug (n=10). PA 1% was administered orally (7.59±1.39 ml/day); while scopolamine (2 mg/Kg), galantamine (1 mg/Kg) and nicotine (0.1 mg/Kg) were administered intraperitoneally. Behavioral tests included: anxiety maze (AM), open field (OF) and marble burying (MB). Habituation cognitive behavior was evaluated in 4 sessions, one week each session. PA had anxiolytic and antidepressant effect effect in AM, combined with nicotine potentiated an anxiogenic effect in AM, galantamine favored habituation in OF. Scopolamine potentiated the habituation in LA and decreased the obsessive-compulsive behavior in OF. In conclusion; PA had an anxiolytic effect and favored deshabituation, combined with nicotine induced an anxiogenic effect, galantamine favored habituation and scopolamine decreased obsessive-compulsive behavior and favored motor habituation indicated a possible anxiolytic effect.

La Petiveria alliacea (PA) está relacionada con efectos ansiolíticos, antidepresivos y cognitivos. El presente trabajo estudió el efecto de la infusión de PA y drogas colinérgicas sobre la habituación. 40 ratones NMRI machos fueron divididos en 4 grupos: Control (n=10), Control Drogas (n=10), PA (n=10) y PA plus Drogas (n=10). La PA (1%) fue administrada vía oral (7.59±1.39 ml/día); escopolamina (2 mg/Kg), galantamina (1 mg/Kg) y nicotina (0.1 mg/Kg) fueron administrados vía intraperitoneal. Los ensayos conductuales incluyeron: laberinto de ansiedad (LA), campo abierto (CA) y enterramiento aversivo (EA). La habituación fue evaluada en 4 sesiones con duración de una semana cada una. PA mostró un efecto ansiolítico en el LA, combinada con nicotina potenció un efecto ansiogénico en el LA. Galantamina favoreció la habituación en CA, y escopolamina potenció el fenómeno de habituación en LA y disminuyó la conducta obsesivo-compulsiva en CA. En conclusión, la PA mostró un efecto ansiolítico y antidepresivo que potencia la deshabituación, combinada con nicotina indujo un efecto ansiogénico, galantamina favoreció la habituación y escopolamina disminuyó la conducta obsesivo­ compulsiva y favoreció la habituación motora indicando un posible efecto ansiolítico.

Antifungal activity of Gallesia integrifolia fruit essential oil

ABSTRACT Gallesia integrifolia (Phytolaccaceae) is native to Brazil and has a strong alliaceous odor. The objective of this study was to identify the chemical composition of G. integrifolia fruit essential oil and evaluate fungicidal activity against the main food-borne diseases and food spoilage fungi. The essential oil was extracted by hydrodistillation and identified by GC-MS. From 35 identified compounds, 68% belonged to the organosulfur class. The major compounds were dimethyl trisulfide (15.49%), 2,8-dithianonane (52.63%) and lenthionine (14.69%). The utilized fungi were Aspergillus fumigatus, Aspergillus niger, Aspergillus ochraceus, Aspergillus versicolor, Penicillium funiculosum, Penicillium ochrochloron, Penicillium verrucosum var. cyclopium, and Trichoderma viride. Minimal fungicidal concentration for the essential oil varied from 0.02 to 0.18 mg/mL and bifonazole and ketoconazole controls ranged from 0.20 to 3.50 mg/mL. The lower concentration of the essential oil was able to control P. ochrochloron, A. fumigatus, A. versicolor, A. ochraceus and T. viride. This study shows a high fungicidal activity of G. integrifolia fruit essential oil and can support future applications by reducing the use of synthetic fungicides.

Efeitos do sombreamento na anatomia foliar de Gallesia integrifolia (Spreng) Harms e Schinnus terebinthifolius Raddi / Effects of shading on leaf anatomy of Gallesia integrifolia (Spreng) Harms and Schinnus terebinthifolius Raddi

Realizou-se estudo com o objetivo de analisar as alterações na anatomia foliar de Gallesia integrifolia (Spreng) Harms e Schinnus terebinthifolius Raddi quando cultivadas em ambientes de sombra moderada ou densa, simulando as condições naturais encontradas em sistemas agroflorestais tradicionais do sul da Bahia, Brasil. Plantas das duas espécies, com aproximadamente um ano de idade, foram cultivadas em casa de vegetação sob quatro níveis de sombreamento (25%, 17%, 10% e 5%). Estudos anatômicos do limbo foliar foram realizados a partir de material incluído em parafina e seccionado em micrótomo rotativo. Os diferentes níveis de sombreamento ocasionaram alterações na estrutura do mesofilo de ambas as espécies, com diferenças significativas na espessura do parênquima paliçádico, limbo foliar, e densidade estomática. Nas condições em que o experimento foi realizado os resultados obtidos indicaram que G. integrifolia apresenta maior capacidade de aclimatação a ambientes de sombra moderada e densa do que S. terebinthifolius, sendo mais indicada para o cultivo em sistemas agroflorestais pré-estabelecidos.

This study was conducted with the aim of analyzing the changes in the leaf anatomy of Gallesia integrifolia (Spreng) Harms and Schinnus terebinthifolius Raddi when grown in environments with moderate to dense shade, simulating the natural conditions found in traditional agroforestry systems in southern Bahia, Brazil. Plants of both species, being approximately one year-old, were grown at four irradiance levels (25%, 17%, 10% and 5%) in a greenhouse. Leaf anatomical studies were made from material embedded in paraffin and sectioned on a rotary microtome. The different irradiance levels caused a shift in the mesophyll anatomy of both species, with significant differences in palisade parenchyma thickness and leaf stomatal density. In the conditions of this experiment, the results indicated that G. integrifolia has a higher capacity for acclimatization to moderate to dense shade than S. terebinthifolius, and it is thus more suitable for cultivation in pre-established agroforestry systems.

Alteraciones morfológicas en el tracto respiratorio de ratas Wistar inducidas por vapores de la raíz de hierba del zorrillo (Petiveria alliacea) del suroeste de México / Morphologic alterations in the respiratory tract of Wistar rats induced by steams of the root of hierba del zorrillo (Petiveria alliacea) from southwest of Mexico

Petiveria alliacea, es conocida con diferentes nombres según el lugar donde se le encuentre. Estudios con hojas, tallo, raíz o extractos describen múltiples usos medicinales. Sin embargo, son pocos los que describen efectos tóxicos. En este estudio se evaluó el efecto morfológico de los vapores de la raíz de P. alliacea sobre el tracto respiratorio de ratas Wistar. Se emplearon 15 ratas divididas en 5 grupos (n=3): control absoluto, 0, 5, 15 y 30 minutos post-exposición (grupos I-V, respectivamente). Las ratas se sacrificaron y se colectaron muestras representativas del tracto respiratorio que posteriormente se procesaron por la técnica histológica convencional, hasta su inclusión en bloques de parafina. Los cortes histológicos se tiñeron con H-E, tricrómico de Masson y azul de toluidina. En tráquea, bronquiolos y pulmón de las ratas de los grupos I y II se observó una histología normal. En la tráquea de los grupos III, IV y V se identificaron áreas variables de hiperplasia en el epitelio, zonas desprovistas de cilios, signos de aumento en la secreción de las células caliciformes y áreas desprovistas de epitelio que se incrementaron con el tiempo. En la lámina propia se observó congestión vascular e infiltrado mononuclear que incrementó con el tiempo. En los bronquiolos de los grupos III y IV se observó activación de las células de Clara, áreas desprovistas de epitelio, y células mononucleares en la luz bronquiolar. En el grupo V se observaron características histológicas normales. En pulmón de los grupos III y IV se identificó engrosamiento de tabiques alveolares, incremento de las fibras de colágena, congestión y extravasación capilar, además de exudado intralveolar. En el grupo V se observó aparente reversión de algunas alteraciones morfológicas de los grupos previos, aunque otras alteraciones persisten. No se observaron diferencias en el número de las células cebadas.

Petiveria alliacea, is known with different names depending of the place where it is found. Studies with leaves, stem, root or extracts, describe multiple medicinal uses. However, there are few reports that describe toxic effects. This study assessed the morphologic effect induced by steams of the root of P. alliacea on the respiratory tract of Wistar rats. We used 15 rats divided in 5 groups (n= 3): absolute control, 0, 5, 15 and 30 minutes post-exposure (I-V groups, respectively). The rats were sacrificed; representative samples of the respiratory tract were collected, and were processed by conventional histological technique until their inclusion in paraffin blocks. Histological sections were stained with H & E, Masson trichrome and toluidine blue stain. A normal histology was observed in trachea, bronchioles and lungs of rats in groups I and II. In trachea in groups III, IV and V, areas with variable hyperplasia in the epithelium, cells without cilia, increase in the secretion of goblet cells, and areas without epithelium were observed. These morphologic alterations were increased with time. In the lamina propia vascular congestion and mononuclear infiltrate were observed, also increasing with time. In bronchioles in groups III and IV, it was noted activation of Clara cells, areas without epithelium, and mononuclear cells in the bronchiolar light. In group V normal histology was observed. In lung in groups III and IV thickening of interalveolar septa, increase of collagen fibers, capillary congestion, extravasation, and exudates were present. Also was observed an apparent reversion of morphologic alterations of previous groups, but other alterations persist. No difference in mast cells number was observed.

In vitro antimicrobial activity of total extracts of the leaves of Petiveria alliacea L. (Anamu)

The antimicrobial activity of 13 total extracts was evaluated, 10 soft extracts (B) and 3 blended extracts (E) prepared from dry and fresh leaves of Petiveria alliacea L. Various solvents were used for their preparation: hydroalcoholic solution at 30%, 80% and isopropyl alcohol. The antimicrobial effect of the extracts was tested by means of the method of Kirby-Bauer, using four bacterial strains from the ATCC collection (Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Pseudomonas aeruginosa) and a leveduriform fungus (Candida albicans). The following quality control parameters were determined for most active extracts: physical, physical-chemical and chemical parameters. The results were: nine extracts showed antibacterial activity, being the most concentrated (B8 and E3), the ones with the highest activity in the presence of the bacteria tested; the effect of blended extracts (E1, E2 and E3) was greater in the presence of P. aeruginosa. Blended extracts are considered more potent and active than soft extracts. No antifungal activity was obtained for both types of extracts. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined for both extracts, with the following results: MIC-soft extracts (>100 mg/mL), blended extracts (>50 mg/mL); MBC-soft extracts (≥400 mg/mL), blended extracts (≥200 mg/mL) based on fresh leaves.

Avaliou-se a atividade antimicrobiana de 13 extratos totais, 10 brandos (B) e 3 batidos (E) elaborados a partir da folha seca e fresca da Petiveria alliacea L. Para sua preparação se empregaram vários solventes: solução hidroalcoólica 30%, 80% e álcool isopropílico. O efeito antimicrobiano dos extratos foi comprovado pelo método do Kirby-Bauer, usando para isso quatro cepas bacterianas da coleção ATCC (Escherichia coli, Staphylococcus aureus, Enterococcus faecalis e Pseudomonas aeruginosa) e um fungo leveduriforme (Candida albicans). Determinaram-se parâmetros de controle da qualidade físicos, físico-químicos e químicos dos extratos mais ativos. Obtiveram-se como resultados: que 9 extratos mostraram atividade antibacteriana, sendo os mais concentrados (B8 e E3) os de maior espectro de ação frente às bactérias provadas; o efeito dos extratos batidos (E1, E2 e E3) foi maior frente a P. aeruginosa; os extratos batidos se consideram mais ativos que os brandos e não se obteve atividade antifúngica caso algum. Determinou-se a Concentração Mínima Inhibitoria e Bactericida para ambos os extratos, resultando a primeira para os extratos brandos maior a 100 mg/mL e para as batidos maior a 50 mg/mL; a segunda para os extratos brandos maior ou igual a 400 mg/mL e para as batidos maior ou igual a 200 mg/mL, em relação à droga fresca.

Efecto protector de Petiveria alliacea L. (Anamú) sobre la inmunosupresión inducida por 5-fluoruracilo en ratones Balb/c / Protecting effect of Petiveria Alliacea (Anamu) on the immunosuppression induced by 5-fluoruracil in Balb/c mice

A preclinical study was carried out to determinate the protective properties of Petiveria alliacea Linn on 5-Fluoruracilo (5-FU)-immunosuppressed animals, a cytostatic drug often used in cancer treatment. Were use five groups of female Balb/c mice (5 mice/group).Two groups were treated with 400 and 1200 mg/kg of P. alliacea leaves and stems powder respectively, and a third group was treated with carboxymethyl cellulose as vehicle. Two additional control groups were set up: a 5-FU treated group, as immunosuppression control, and a NaCl solution (0.9 percent treated group. Animals were treated daily for five days and then a unique dose of 150 mg/kg of 5-FU was administered and the treatment continued for another four days. At termination blood and tissue samples were collected for leukocyte total count, analysis of bone marrow cellularity, thymus weight and total IgG antibody forming cells. Our results show that the group treated with the highest dose of P. alliacea, was less affected by 5-FU-induced immunosupresion compared with the other treated groups. The results derived from this study suggest that P. alliacea, a medicinal plant product, could be used in patients under antineoplasic regimens to avoid the deleterious adverse effects of the immunosuppressive drugs.

Se realizó un estudio preclínico para la determinación de las propiedades protectoras de la planta Petiveria alliacea Linn sobre la inmunosupresión inducida por la droga citostática 5- Fluoruracilo (5-FU), la cual se utiliza muy frecuentemente en la terapia contra el cáncer. Se utilizaron cinco grupos de ratones hembras Balb/c (5 ratones por grupo) que incluyeron dos grupos de tratamiento con dos niveles de dosis del polvo de las hojas y tallos de la planta: 400 y 1200 mg/kg, así como grupos controles con solución de NACl y con el vehículo (solución de carboximetil celulosa) por vía oral, aplicados durante 5 días, luego una administración única de 150 mg/kg de 5-FU y la continuación del tratamiento en los restantes 5 días. En las variables: conteo global y diferencial de leucocitos, celularidad de la médula ósea, peso del timo y Células Formadoras de Anticuerpos (CFA) IgG totales, se pudo observar que el grupo de mayor dosis de P. alliacea tuvo una menor afectación por la inmunosupresión inducida por 5-FU, en comparación con el resto de los grupos tratados. Estos resultados apoyan el uso de formulaciones de esta planta en pacientes que reciben tratamientos antineoplásicos para la protección contra la inmunosupresión.

Histopatologia da interação de Bacillus thuringiensis e extratos vegetais no intestino médio de Spodoptera frugiperda (Lepidoptera: Noctuidae) / Histopathology of the interaction of Bacillus thuringiensis and plant extracts in the midgut of Spodoptera frugiperda (Lepidoptera: Noctuidae)

Plantas são fontes naturais de substâncias inseticidas, já que podem ser produzidas pelo vegetal em resposta a ataques de insetos podendo representar uma alternativa no manejo de insetos-praga, como Spodoptera frugiperda. Dessa forma, o presente trabalho objetivou a análise histopatológica do intestino médio de lagartas de S. frugiperda, após a ingestão dos extratos obtidos por maceração e infusão de Petiveria alliacea. Zingiber officinale. Cymbopogon citratus. Malva silvestris. Baccharis genistelloides e Ruta graveolens, assim como a associação desses extratos com Bacillus thuringiensis aizawai. As lagartas foram tratadas in vivo com cada extrato e a bactéria, mais a associação desses extratos com B. thuringiensis e após, uma reação de cinética entre 3 e 27 horas, foram fixadas para o preparo de cortes histológicos, os quais foram corados e analisados comparativamente às testemunhas em microscopia óptica. Os resultados mostraram mudanças na histologia do intestino médio das lagartas de S. frugiperda, 3 horas após a aplicação dos tratamentos à base de alliacea. Z. officinale. C. citratus e M. silvestris, enquanto que para B. genistelloides e R. graveolens só foram observados alterações após 6h. Na interação dos extratos com B. thuringiensis observou-se alterações nas microvilosidades, desorganização do intestino médio e a hipertrofia das células epiteliais que projetaram-se para o lúmen. Os resultados desse trabalho mostram que o efeito histopatológico de Z. officinale. M. silvestris. R. graveolens e B. genistelloides, foram mais ativos quando comparados aos extratos de P. alliacea e C. citratus, os quais apresentaram uma interação positiva com B. thuringiensis.

The natural insecticidal substances produced by plants in response to insect attack may represent an alternative for the management of pest insects such as Spodoptera frugiperda. This study was therefore aimed to histopathologically analyze the midgut of larvae of S. frugiperda, after ingestion of extracts obtained by maceration and infusion of Petiveria alliacea. Zingiber officinale. Cymbopogon citratus. Malva silvestris. Baccharis genistelloides and Ruta graveolens, and the association of these extracts with Bacillus thuringiensis aizawai. The larvae were treated in vivo with each extract, B. thuringiensis, and the combination of these extracts with the bacteria, and after a reaction kinetics of between 3 to 27 hours they were fixed for the preparation of histological sections, which were stained and analyzed in comparison to the controls with light microscopy. The results showed changes in the histology of the midgut of the larvae of S. frugiperda, 3 hours after application of treatments based on P. alliacea. Z. officinale. C. citratus and M. silvestris, while for B. genistelloides and R. graveolens changes were observed only after 6h. In the interaction of extracts with B. thuringiensis the analysis revealed changes in microvilli, disorganization of the midgut and hypertrophy of epithelial cells that projected into the lumen. The findings show that the histopathological effects of Z. officinale. M. silvestris. R. graveolens and B. genistelloides were more active when compared to extracts of P. alliacea and C. citratus, which showed a positive interaction with B. thuringiensis.

Analysis of fetal and placental development in rats after administration of hydroalcoholic extract from the root of Petiveria alliacea L. (Phytolaccaceae) / Análisis del desarrollo fetal y placenta en ratas después de la administración de extracto hidroalcohólico de la raíz de Petiveria alliacea L. (Phytolaccaceae)

We investigated the effect of hydroalcoholic extract the Petiveria alliacea root on fetal and placental development female rats pregnancy. Thirty female rats were mated and divided in groups: I - rats treated with placebo and sacrificed on the 7th day; II - rats treated with hydroalcoholic extract and sacrificed on the 7th day; III - rats treated with placebo and sacrificed on the 14th day; IV - rats treated with hydroalcoholic extract and sacrificed on the 14th day; V - rats treated with placebo for analysis of the offspring; VI - rats treated with hydroalcoholic extract for analysis of the offspring. The extract was administered by gavage in the dosage of 18mg/kg body wt., in 5th day of pregnancy. The implantation sites and placenta were fixed in Bouin and processed for the embedding in paraffin. The offspring were weighed, measured and counted. The hydroalcoholic extract of the root of the P. alliacea causes significant reduction in the number of implantation sites, but do not cause histological alterations in these sites and placenta. No alterations in the number, length and weight have been observed in offspring. Our results suggest that the hydroalcoholic extract from the P. alliacea in the dosage of 18mg/kg body wt., administered to female rats, on the 5th day of pregnancy, only cause a retard in the implantation process.

Se investigaron los efectos de la extracto hidroalcohólico de la raíz Petiveria alliacea sobre el desarrollo fetal y placentario en ratas preñadas. Treinta ratas hembras fueron criadas y divididas en grupos: I- Ratas tratadas con placebo y sacrificado en el día 7; II- Ratas tratadas con el extracto hidroalcohólico y sacrificadas en el día 7; III- ratas tratadas con placebo y sacrificado el día 14; IV- ratas tratadas con el extracto hidroalcohólico y sacrificaron el día 14; V- ratas tratadas con placebo para el análisis de las crías; VI- ratas tratadas con el extracto hidroalcohólico para el análisis de las crías. El extracto se administró por sonda en dosis de 18mg/kg de peso corporal, en el 5to día de preñez. Los sítios de implantación y la placenta se fijaron en Bouin y se procesaron para la inclusión en parafina. Las crías fueron pesadas, medidas y contadas. El extracto hidroalcohólico de la raíz de P. alliacea provocó una reducción significativa en el número de sitios de implantación, pero no causaron alteraciones histológicas en estos sitios y la placenta. No se observaron alteraciones en el número, longitud y peso de las crías. Nuestros resultados sugieren que el extracto hidroalcohólico de P. alliacea en la dosis de 18mg/kg de peso corporal, administrado a ratas hembras en el 5to día de preñez, sólo causó retardo en el proceso de implantación.

Implications of dibenzyl trisulphide for disease treatment based on its mode of action / Implicaciones del trisulfuro de dibencilo para el tratamiento de enfermedades a partir de su modo de acción

Studies conducted on the secondary metabolite (natural product), dibenzyl trisulphide (DTS), which was isolated from the sub-tropical shrub Petiveria alliacea (guinea hen weed, anamu) [Phytolaccaceae] have shown tremendous pharmaceutical promise as a drug prototype. This is now reflected in the development of the broad spectrum anti-cancer molecule, fluorapacin (bis(4-fluorobenzyl) trisulphide) which has an excellent safety profile. The mode of action elucidated for DTS is the mitogen activated protein extracellular regulated kinases 1 and 2 (MAPKinases ERK 1 and ERK 2). The MAPKinase signal transduction biochemical pathways are important in the regulation of a wide range of cellular processes which are important in disease establishment. These processes include: cancer cell proliferation, nerve repair, memory enhancement, autoimmune diseases, which are linked to thymic cell involution and bone marrow functions, cerebrovascular and cardiovascular diseases. In addition to the MAPkinase signal transduction mode of action, DTS also prevents the denaturation of serum albumin which is a feature of nonsteroidal anti-inflammatory drugs, thus supporting the molecule's possible role in the treatment of inflammatory ageing diseases.

Los estudios realizados sobre el metabolito secundario (producto natural), trisulfuro de dibencilo (TSD), que fue aislado del arbusto subtropical Petiveria alliacea (hierba de guinea, anamú) [Phytolaccaceae] muestran que se trata de una tremenda promesa farmacéutica como prototipo de droga. Esto se refleja actualmente en el desarrollo de la molécula anticancerígena de amplio espectro, la fluorapacina (bis (4-fluorobencilo) trisulfuro) que posee un excelente perfil de seguridad. El modo de acción para el TSD se explica partiendo de las proteínas quinasas 1 y 2 activadas por mitógenos y reguladas por señales extracelulares (Quinasas MAP ERK 1 y ERK 2). Las vías bioquímicas de transducción de la señal de la quinasa MAP, son importantes en la regulación de una amplia variedad de procesos celulares, importantes a la hora de determinar una enfermedad. Dichos procesos comprenden: la proliferación de la célula cancerosa, la reparación de nervios, el mejoramiento de la memoria, y las enfermedades autoinmunes, vinculadas con la involución tímica y las funciones de la médula, las enfermedades cerebrovasculares y cardiovasculares. Además del modo de acción de las transducción de señales de la quinasa MAP, el TSD previene también la desnaturalización de la albúmina sérica, lo cual constituye una característica de las drogas anti-inflamatorias no esferoidales, apoyando así el posible papel de las moléculas en el tratamiento de las enfermedades inflamatorias en el proceso de envejecimiento.

Subchronic and acute preclinic toxicity and some pharmacological effects of the water extract from leaves of Petiveria alliacea (Phytolaccaceae)

We tested the effects of the aqueous extract of Petiveria alliacea leaves on acute and sub-chronic toxicity, hematocrit and blood glucose level and intestinal motility of male albino NGP mice of 20 to 25 g mean weight. Treatments were in all cases doses of 1,000 and 2,000 mg/kg animal weight and a control treatment with 0.5 ml distilled water, using 10 animals per treatment and administered orally every day (5 days per week). Experimental periods were 18 and 70 days for acute and sub chronic toxicity, respectively. No mortality nor any toxicity signs could be observed. A slight but significant increase in the glucose levels during the first three weeks was observed with the 1,000 mg/kg dose but not for the higher 2,000 mg/kg dose. After administering the doses once after a starving period of six hours, no significant differences in intestinal motility could be found.

Se evaluaron los efectos del estracto acuoso de las hojas de Petiveria alliacea, en la toxicidad aguda y toxicidad subcrónica, hematocritos, niveles de glucosa en la sangre y motilidad intestinal del ratón macho albino NGP, con un peso promedio de 20 a 25g. En todos los casos los tratamientos fueron dosis de 1 000 y 2 000 mg/kg de peso del animal y un tratamiento control con 0.5 ml de agua destilada, usando 10 animales por tratamiento y administrado oralmente cinco días por semana. Los períodos experimentales fueron de 18 y 70 días para toxicidad aguda y toxicidad subcrónica, respectivamente. No se observaron signos de mortalidad ni de toxidad en ambas pruebas. Con la dosis de 1 000 mg/kg hubo un leve pero significativo incremento en los niveles de glucosa durante las primeras tres semanas, pero no con la dosis más alta de 2 000 mg/ kg. Después de administrar las dosis luego de un período de hambre de seis horas, no se encontraron diferencias significativas en la motilidad intestinal.