Selective cytotoxic effect of Annona muricata L. in HCC1954 (HER2+) breast cancer cells / Efecto citotóxico selectivo de Annona muricata L. en células HCC1954 (HER2+) de cáncer de mama

Annona muricata Linn. (Annonaceae) is a tropical plant with multiple beneficial health effects including anticancer properties. In breast cancer patients, overexpression of the HER2 oncoprotein corresponds to a poor prognosis, thus the main purpose of this study was to evaluate the cytotoxicity of ethanolic extracts from dried and fresh leaf of A. muricata on HER2+ breast cancer cells. MTT assays were performed and IC50 determined in HCC1954 (HER2+) cells, as well as in MCF7 (HER-) and peripheral blood mononuclear cells (PBMC) used as controls. Total polyphenol content evaluation and phytochemical screening were also performed. The cytotoxic effect of A. muricata extracts (125-1000 µg/mL) was dose-dependent and cell-type specific. The extracts exhibited higher cytotoxicity against HCC1954 than MCF7 cells, but weak toxicity against PBMC. This is the first report of the cytotoxic effect of A. muricata on HCC1954 cells, highlighting its potential for treating anti-estrogen-resistant breast cancers and low toxicity against PBMC.

Annona muricata Linn. (Annonaceae) es una planta tropical con múltiples efectos benéficos en la salud incluyendo propiedades antitumorales. En pacientes con cáncer de mama la sobreexpresión del oncogen HER2 corresponde a un mal pronóstico, por lo que el objetivo principal de este estudio fue evaluar la citotoxicidad de extractos etanólicos de hojas secas y frescas de A. muricata en células tumorales de mama HER2+. Se aplicaron pruebas de MTT y se determinaron IC50en células HCC1954 (HER2+); se utilizaron células MCF7 (HER-) y células mononucleares de sangre periférica (PBMC) como control. Se valoró también el contenido en polifenoles totales, y se realizó un tamizaje fitoquímico. El efecto citotóxico de los extractos de A. muricata (125-1000 µg/mL) fue dosis-dependiente y específico para cada tipo celular. Los extractos presentaron mayor actividad citotóxica contra HCC1954 en comparación con MCF7 y baja toxicidad contra PBMC. Este es el primer reporte del efecto citotóxico de A. muricata en HCC1954 y destaca su potencial terapéutico para tratamiento de cáncer de mama resistentes a antiestrógeno y baja citotoxicidad contra PBMC.

Actividad biológica de los aceites de semillas de Annona muricatay A. cherimola / Biological activity of Annona muricata and A. cherimola seed oils

Vegetable extracts have become important raw materials for food, pharmaceutical and cosmetic industries because of their biological potential. The objective of this study was to assess the biological activity of vegetable oils (VOs) extracted from Annona muricata and A. cherimola. Antibacterial activity was determined by plaque microdilution. The assessment of hemolytic inhibition and morphological alterations was performed in erythrocyte cultures by spectrophotometry and microscopy, respectively. Neutrophils were used to analyze both cytotoxicity by the trypan blue exclusion method and the effect on gelatinase granule release (MMP9) via zymography. Whereas VOs showed a mild antibacterial activity (900 µL/mL) on five ATCC bacterial strains, they had no effect on multi-resistant bacteria. In addition, VOs inhibited hydrogen peroxide induced hemolysis and did not cause erythrocyte cell abnormalities. Cytotoxicity was not detected in neutrophils and VOs were able to stimulate MMP9 release. These results support their potential use by the food and cosmetic industries due to their antioxidant, non-cytotoxic, and slight antibacterial capacities.

Los extractos vegetales adquieren importancia en la industria alimentaria, farmacéutica y cosmética, por su potencial biológico. El objetivo de este estudio fue evaluar actividad biológica de los Aceites Vegetales (AV) de semillas de Annona muricata y A. cherimola. La actividad antibacteriana se determinó mediante microdilución en placa; en cultivo de eritrocitos se evaluó inhibición hemolítica por espectrofotometría y alteraciones morfológicas por recuento microscópico; en neutrófilos se evaluó citotoxicidad por método de exclusión con azul de tripán, y el efecto sobre liberación de gránulos de gelatinasa (MMP9) mediante zimografía. Los AV presentaron actividad antibacteriana leve (900 µL/mL) en cinco cepas ATCC, pero no en bacterias multirresistentes; inhibieron la hemolisis inducida por peróxido de hidrogeno; no generaron deformaciones eritrocitarias; no se evidenció citotoxicidad en neutrófilos y estimularon la liberación de MMP9. Los resultados podrían sustentar el uso potencial de estos AV en la industria alimenticia o cosmética, gracias a su capacidad antioxidante, no citotóxica y levemente antibacteriana.

Inheritance of seedlessness and the molecular characterization of the INO gene in Annonaceae / Herança da ausência de sementes e a caracterização molecular do gene INO em Annonaceae

The inheritance of the seedless fruit characteristic of Annona squamosa has not yet been explained. Molecular techniques may aid breeding programs, mainly in the assisted selection of the target gene. The INO gene may be related to seed development in these fruits. The objective of the present paper was to investigate the inheritance of seedlessness in the 'Brazilian seedless' sugar apple and INO gene conservation in Annona squamosa and Annona cherimola x Annona squamosa genotypes by assessing their homology with the INO database genes. The F1 generation was obtained by crossing the mutant 'Brazilian seedless' (male genitor) (P1) with the wild-type A. squamosa with seeds (M1 and M2, female genitors). The INO gene was studied in mutant and wild-type A. squamosa (P1, M1, M2 and M3) and in the Gefner atemoya (A. cherimola x A. squamosa) (M4) cultivar. The DNA was extracted from young leaves, and four sets of specific primers flanking the INO gene were amplified. The seedless characteristic was identified as stenospermatic in the fruits of parental P1, suggesting monogenic inheritance with complete dominance. High sequence similarity of the INO gene amplifications in the sugar apple accessions (M1, M2, M3) and the atemoya cultivar Gefner (M4) reinforces the hypothesis of their conservation.

A herança da característica de fruto sem sementes de Annona squamosa ainda não foi esclarecida. Técnicas moleculares podem auxiliar em programas de melhoramento, principalmente na seleção assistida do gene de interesse. O gene INO pode estar relacionado ao desenvolvimento da semente dessas frutas. O objetivo foi investigar a herança da ausência de sementes em Annona squamosa e a conservação do gene INO nos genótipos Annona squamosa e Annona cherimola x Annona squamosa avaliando sua homologia com banco de dados de genes INO. A geração F1 foi obtida pelo cruzamento do mutante 'Brazilian seedless' (genitor masculino) (P1) com o tipo selvagem com sementes (A. squamosa) (M1 e M2, genitores femininos). O gene INO foi estudado em A. squamosa, mutante e selvagem (P1, M1, M2 e M3) e na cultivar Gefner atemoya (A. cherimola x A. squamosa) (M4). O DNA foi extraído de folhas jovens, e quatro conjuntos de primers específicos flanqueando o gene INO foram amplificados. A característica sem sementes foi identificada como estenospermática nos frutos do parental P1, sugerindo herança monogênica com dominância completa. A alta similaridade de sequência das amplificações do gene INO nos acessos de pinha (M1, M2, M3) e na cultivar de atemóia Gefner (M4) reforça a hipótese de sua conservação.

Inheritance of seedlessness and the molecular characterization of the INO gene in Annonaceae / Herança da ausência de sementes e a caracterização molecular do gene INO em Annonaceae

Abstract The inheritance of the seedless fruit characteristic of Annona squamosa has not yet been explained. Molecular techniques may aid breeding programs, mainly in the assisted selection of the target gene. The INO gene may be related to seed development in these fruits. The objective of the present paper was to investigate the inheritance of seedlessness in the 'Brazilian seedless' sugar apple and INO gene conservation in Annona squamosa and Annona cherimola x Annona squamosa genotypes by assessing their homology with the INO database genes. The F1 generation was obtained by crossing the mutant 'Brazilian seedless' (male genitor) (P1) with the wild-type A. squamosa with seeds (M1 and M2, female genitors). The INO gene was studied in mutant and wild-type A. squamosa (P1, M1, M2 and M3) and in the Gefner atemoya (A. cherimola x A. squamosa) (M4) cultivar. The DNA was extracted from young leaves, and four sets of specific primers flanking the INO gene were amplified. The seedless characteristic was identified as stenospermatic in the fruits of parental P1, suggesting monogenic inheritance with complete dominance. High sequence similarity of the INO gene amplifications in the sugar apple accessions (M1, M2, M3) and the atemoya cultivar Gefner (M4) reinforces the hypothesis of their conservation.

Resumo A herança da característica de fruto sem sementes de Annona squamosa ainda não foi esclarecida. Técnicas moleculares podem auxiliar em programas de melhoramento, principalmente na seleção assistida do gene de interesse. O gene INO pode estar relacionado ao desenvolvimento da semente dessas frutas. O objetivo foi investigar a herança da ausência de sementes em Annona squamosa e a conservação do gene INO nos genótipos Annona squamosa e Annona cherimola x Annona squamosa avaliando sua homologia com banco de dados de genes INO. A geração F1 foi obtida pelo cruzamento do mutante 'Brazilian seedless' (genitor masculino) (P1) com o tipo selvagem com sementes (A. squamosa) (M1 e M2, genitores femininos). O gene INO foi estudado em A. squamosa, mutante e selvagem (P1, M1, M2 e M3) e na cultivar Gefner atemoya (A. cherimola x A. squamosa) (M4). O DNA foi extraído de folhas jovens, e quatro conjuntos de primers específicos flanqueando o gene INO foram amplificados. A característica sem sementes foi identificada como estenospermática nos frutos do parental P1, sugerindo herança monogênica com dominância completa. A alta similaridade de sequência das amplificações do gene INO nos acessos de pinha (M1, M2, M3) e na cultivar de atemóia Gefner (M4) reforça a hipótese de sua conservação.

Natural parasitism in fruit fly (Diptera: Tephritidae) and interaction with wild hosts surrounding apple orchards adjacent to Atlantic Forest fragments in Paraná State, Brazil / Parasitismo natural de mosca-das-frutas (Diptera: Tephritidae) e interação com hospedeiros silvestres em pomares de macieira adjacentes a fragmentos de Mata Atlântica no Estado do Paraná, Brasil

The South American fruit fly, Anastrepha fraterculus (Wiedemann, 1830) (Diptera: Tephritidae), is an important pest in the subtropical region of Brazil. This insect has tritrophic relation between wild fruits and parasitoids and is associated with apple (Malus domestica Borkh.) orchards adjacent to the Atlantic Forest in Paraná. We thus investigated the degree of infestation of the fruit fly and natural parasitism in wild and cultivated fruits surrounding apple orchards. For this purpose, we collected fruits of Acca sellowiana (Berg.) Burret, Campomanesia xanthocarpa (Mart), Eugenia uniflora L., Eugenia pyriformis Cambessèdes, Psidium cattleianum Sabine, Psidium guajava (L.), Annona neosericea Rainer and Eriobotrya japonica (Thumb) in apple orchards adjacent to the Atlantic Forest located in Campo do Tenente, Lapa and Porto Amazonas counties. In total, we collected 18,289 fruits during four growing years. The occurrence of A. fraterculus depends on the susceptible period of apple fruits. A. sellowiana and P. cattleianum were considered primary fruit fly multipliers and P. guajava was secondary, all occurring after the apple harvest (IS period). The group of parasitoids with A. fraterculus was Aganaspis pelleranoi (Brèthes, 1924) (Hymenoptera: Figitidae), Opius bellus (Gahan, 1930), Doryctobracon areolatus (Szépligeti, 1911) and Doryctobracon brasiliensis (Szépligeti, 1911) (Hymenoptera: Braconidae) all of which are first records in the Atlantic Forest in Paraná. First record of O. bellus occurring in the State of Paraná, as well as, first record of the tritrophic association between host plant A. neosericea, parasitoids D. areolatus and O. bellus and fruit fly A. fraterculus. The host P. cattleianum stood out among the Myrtaceae species in regard to the high diversity of parasitoid species (81% of parasitoids). The total number of Figitidae species (76.5%) was higher [...].

Mosca-das-frutas sul-americana, Anastrepha fraterculus (Wiedemann, 1830) (Diptera: Tephritidae), é uma importante praga da região subtropical do Brasil. Este inseto tem relação tritrófico entre frutos silvestres e parasitoides e está associado a pomares de macieiras (Malus domestica Borkh.) adjacentes à Mata Atlântica no Paraná. Assim, investigamos o grau de infestação da mosca-das-frutas e o parasitismo natural em frutas silvestres e cultivadas ao redor de pomares de maçã. Para tanto, foram coletados frutos de Acca sellowiana (Berg.) Burret, Campomanesia xanthocarpa (Mart), Eugenia uniflora L., Eugenia pyriformis Cambessèdes, Psidium cattleianum Sabine, Psidium guajava (L.), Annona neosericea Rainer e Eriobotrya japonica (Thumb) em pomares de maçã adjacentes à Mata Atlântica localizados nos municípios de Campo do Tenente, Lapa e Porto Amazonas. No total, coletamos 18.289 frutos durante quatro anos de cultivo. A ocorrência de A. fraterculus depende do período de suscetibilidade dos frutos da maçã. A. sellowiana e P. cattleianum foram considerados multiplicadores primários de mosca-das-frutas e P. guajava foi secundário, todos ocorrendo após a colheita da maçã (período IS). Os parasitóides a associados a A. fraterculus foram Aganaspis pelleranoi (Brèthes, 1924) (Hymenoptera: Figitidae), Opius bellus (Gahan, 1930), Doryctobracon areolatus (Szépligeti, 1911) e Doryctobracon brasiliensis (Szépligeti, 1911) (Hymenoptera: Braconidae), todos os quais são primeiros registros na Mata Atlântica no Paraná. Primeiro registro de O. bellus ocorrendo no Estado do Paraná, assim como, primeiro registro da associação tritrófica entre o hospedeiro A. neosericea, parasitoides D. areolatus e O. bellus e mosca-das-frutas A. fraterculus. O hospedeiro P. cattleianum se destacou entre as espécies de Myrtaceae pela alta diversidade de parasitóides associados (81% dos parasitóides). O número total de espécies de Figitidae (76,5%) foi superior [...].

Artificial pollination and use of growth regulators in atemoya 'Gefner' fruits

The objective was to study the effectiveness of the growth regulator (ANA + GA3) associated or not to the application of adjuvant and artificial pollination in 'Gefner' atemoya. The experiment was conducted in the experimental orchard at Florida's Tropical Research and Education Center. The experimental design was in a randomized block, with 14 treatments, 10 repetitions and 3 flowers per plot. The highest percentages of fixed fruits were obtained with hand pollination ­ HP and 450 NAA + 1250 GA3 mg L-1 + adjuvant and HP. The use of hand pollination for 'Gefner' atemoya tree proved to be the most efficient method so far. Applying growth regulator without artificial pollination produces parthenocarpic fruits, however with high rate of abortions, and small fruits. Growth regulators together with hand pollination produces small and uneven fruits, and cause reduction in the fruits' titratable acidity. The use of adjuvant caused low fixation and toxicity to fruits, and its use is not recommended.

Alternativas de controle do diabetes / Alternatives for diabetes control / Alternativas para el control de la diabetes

O Diabetes desde a antiguidade tem sido uma das maiores causas de morte entre as populações do globo, e segundo a Organização Mundial da Saúde continua assolando nos nossos dias. Apesar das descobertas de tratamentos mais eficazes, a doença vem avançando em progressões assustadoras atualmente, com projeções preocupantes para a saúde pública. Como estratégia de acompanhamento terapêutico, estatístico direcionado a portadores de diabetes, o Governo Federal lançou o programa HIPERDIA (Hipertensos e Diabéticos), que faz o acompanhamento da evolução da doença e das complicações dos pacientes. E neste sentido, também são utilizadas terapêuticas mais acessíveis como as plantas medicinais. O objetivo desta pesquisa consiste em realizar uma revisão bibliográfica abordando as opções de terapias de controle do diabetes oferecidas no Sistema Único de Saúde e pesquisar fitoterápicos com potencial hipoglicêmico aprovados pela Anvisa. Através de levantamento bibliográfico, foram identificadas oito espécies vegetais utilizadas pela medicina popular no controle do diabetes, sendo estas: Bauhinia Forficata, Syzygium Cumini, Annona Muricata, Cynara Scolymus, Momordica Charantia, Eugenia Uniflora e Baccharis Trimera. Essas plantas do programa, embora tenham comprovação de seu efeito hipoglicêmico e redutores dos sintomas diabéticos, pelas suas propriedades antioxidantes e antiinflamatórias, colabora para uma melhor qualidade de vida aos pacientes.

Since antiquity, Diabetes has been one of the biggest causes of death amon-g populations around the globe, and according to the World Health Organization, it continues to plague our days. Despite discoveries of more effective treatments, the disease is currently advancing in frightening progressions, with worrying projections for public health. As a therapeutic, statistical follow-up strategy aimed at people with diabetes, the Federal Government launched the HIPERDIA (Hypertensive and Diabetic) program, which monitors the evolution of the disease and the complications of patients. And in this sense, more accessible therapies such as medicinal plants are also used. The objective of this research is to carry out a literature review addressing the options for diabetes control therapies offered in the Unified Health System and to search for herbal medicines with hypoglycemic potential approved by Anvisa. Through a bibliographical survey, eight plant species used by folk medicine to control diabetes were identified, namely: Bauhinia Forficata, Syzygium Cumini, Annona Muricata, Cynara Scolymus, Momordica Charantia, Eugenia Uniflora and Bacharis Trimera. These plants in the program, although they have evidence of their hypoglycemic effect and reduce diabetic symptoms, due to their antioxidant and anti-inflammatory properties, contribute to a better quality of life for patients.

La diabetes ha sido desde la antigüedad una de las principales causas de muerte entre las poblaciones del planeta, y según la Organización Mundial de la Salud sigue haciendo estragos en nuestros días. A pesar de los descubrimientos de tratamientos más eficaces, la enfermedad avanza actualmente con una progresión aterradora, con proyecciones preocupantes para la salud pública. Como estrategia de seguimiento terapéutico, estadísticamente dirigida a las personas con diabetes, el Gobierno Federal puso en marcha el programa HIPERDIA (Hipertensión y Diabetes), que controla la evolución de la enfermedad y las complicaciones de los pacientes. En este sentido, también se utilizan terapias más accesibles, como las plantas medicinales. El objetivo de esta investigación es realizar una revisión bibliográfica que aborde las opciones de terapias para el control de la diabetes ofrecidas en el Sistema Único de Salud y buscar fitoterapias con potencial hipoglucemiante aprobadas por Anvisa. Mediante un estudio bibliográfico, se identificaron ocho especies vegetales utilizadas por la medicina popular en el control de la diabetes, a saber: Bauhinia Forficata, Syzygium Cumini, Annona Muricata, Cynara Scolymus, Momordica Charantia, Eugenia Uniflora y Baccharis Trimera. Estas plantas del programa, aunque han demostrado su efecto hipoglucemiante y reductor de los síntomas diabéticos, por sus propiedades antioxidantes y antiinflamatorias, colaboran a una mejor calidad de vida para los pacientes.

Relationship between the toxicity of raw Annona muricata leaves extract and 7, 12-dimethylbenzathracene (DMBA) with histopathological changes in wistar rats / Relación entre la toxicidad del extracto de hojas crudas de Annona muricata y el 7, 12-dimetilbenz antraceno (DMBA) con cambios histopatológicos en ratas Wistar

Abstract Aims: To identify the histopathological alterations in organs of Wistar rats to evaluate toxic effects of use of Annonamuricata Raw Leaf Extract (AMRLE) alone or in association with DMBA. Settings and Design: Sixty female Wistar rats were used, separated into groups and treated with a single dose of 65 mg/kg of DMBA and/or with 50; 100 and 200 mg/kg of AMRLE. Hematoxylin-Eosin (HE) and 1% methylene blue stains were used in the histopathological analysis and quantification of Aberrant Crypts (ACs) and Aberrant Crypt Focus (ACF). Fischer and Kruskal Wallis tests were used in the statistical analysis. Results: The administration of 65 mg/kg of DMBA and/or 50, 100 and 200 mg/kg of AMRLE did not influence weight development. Some histopathological alterations (hepatic steatosis; inflammatory foci in the liver, kidney and lung; pulmonary lymphoid hyperplasia, ectasia and hyperplasia in mammary gland epithelium) and the development of ACs and ACF in the intestinal colon were observed in all groups, except in the group negative control, with no statistical difference between analysed groups. Conclusions: Histopathological alterations and the formation of ACs and ACF did not show a statistically significant difference between the groups analysed. However, although AMRLE has antioxidant effects due to the presence of phenolic components, there was still the formation of some pathological processes that may be related to the isolated toxic action of DMBA and/or associated with other components of AMRLE, since these changes were not seen in the negative control group.

Resumen Objetivos: Identificar las alteraciones histopatológicas en órganos de ratas Wistar para evaluar los efectos tóxicos del uso del Extracto de Hoja Cruda de Annona muricata (AMRLE) solo o en asociación con DMBA. Configuración y diseño: Se utilizaron sesenta ratas hembras Wistar, se separaron en grupos y se trataron con una dosis única de 65 mg/kg de DMBA y/o con 50, 100 y 200 mg/kg de AMRLE. Se utilizaron tinciones de hematoxilina-eosina (HE) y azul de metileno al 1% en el análisis histopatológico y la cuantificación de criptas aberrantes (CA) y focus de criptas aberrantes (FCA). En el análisis estadístico se utilizaron las pruebas de Fischer y Kruskal Wallis. Resultados: La administración de 65 mg/kg de DMBA y/o 50, 100 y 200 mg/kg de AMRLE no influyó en el desarrollo del peso. Se observaron algunas alteraciones histopatológicas (esteatosis hepática; focus inflamatórios en el hígado, riñón y pulmón; hiperplasia, ectasia en epitelio de la glándula mamaria e hiperplasia linfoide pulmonar) y el desarrollo de CA y FCA en el colon intestinal en todos los grupos, excepto en el grupo control negativo, sin diferencias estadísticas entre los grupos analizados. Conclusiones: Las alteraciones histopatológicas y la formación de CA y FCA no mostraron diferencias estadísticamente significativas entre los grupos analizados. Sin embargo, aunque AMRLE tiene efectos antioxidantes debido a la presencia de componentes fenólicos, aún existe la formación de algunos procesos patológicos que pueden estar relacionados con la acción tóxica aislada del DMBA y/o asociados con otros componentes de AMRLE, ya que estos cambios no fueron observados en el grupo control negativo.

Chemical composition, larvicidal and cytotoxic activity of Annona salzmannii (Annonaceae) seed oil

The seed oil of Annona salzmannii A. DC. was analyzed by GC-MS and 1H qNMR, revealing a mixture of unsaturated (80.5%) and saturated (18.7%) fatty acids. Linoleic (45.3%) and oleic (33.5%) acid were the major unsaturated fatty acids identified, while palmitic acid (14.3%) was the major saturated fatty acid. The larvicidal effects of A. salzmannii seed oil were evaluated against third-instar larvae of Aedes aegypti (Linn.). The oil exhibited moderate larvicidal activity, with a LC50 of 569.77 ppm (95% CI = 408.11 to 825.88 ppm). However, when the cytotoxic effects of the oil were evaluated, no expressive antiproliferative effects were observed in tumor cell lines B16-F10 (mouse melanoma), HepG2 (human hepatocellular carcinoma), K562 (human chronic myelocytic leukemia), HL-60 (human promyelocytic leukemia), and non-tumor cell line PBMC (peripheral blood mononuclear cells), with IC50 values > 50 µg·mL-1. This is the first study to evaluate the chemical composition, larvicidal and cytotoxic activity of A. salzmannii seed oil

Isolation of Polyphenols from Soursop Annona muricata L Leaves Using Green Chemistry Techniques and their Anticancer Effect

HIGHLIGHTS Isolate, fractionate and characterize extracts obtained from soursop leaves. Use of emerging green technologies such as microwave-ultrasound hybridization. The extracts contain kaempferol, procyanidins, catechin, and quercetin. The total ethanolic extract demonstrates cytotoxic effect on HeLa cells.

Abstract Cervical cancer is classified as the fourth most common malignancy in women. Natural compounds are a therapeutic alternative in cancer therapy. The aim of the study is to isolate, fractionate, and characterize extracts obtained from soursop leaves (Annona muricata L.) and determine their cytotoxic effect against HeLa cervical cancer cells and non-carcinogenic fibroblast 3T3 cells. The phytochemicals of soursop leaves were extracted through emerging green technologies such as the novel use of microwave-ultrasound hybridization and the use of environmentally friendly solvents (water and ethanol), in addition to the purification of extracts enriched in polyphenols by liquid chromatography with Amberlite XAD-16. Total aqueous and ethanolic extract were purified, as well as the fraction one of each extract. The extracts recovered from soursop leaves contained kaempferol and its isomers, procyanidins, catechin, and quercetin. The viability of the cells was determined with the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. HeLa and 3T3 cells were exposed to concentrations of 25, 50, 75, 100, 150, 200, and 250 ppm of a solution of soursop leaf extract powder. The MTT assay showed that soursop leaf extracts were toxic to both cell lines in general, however, the ethanolic extract at 25 and 50 ppm demonstrated inhibition in cell viability against the HeLa cancer line and low cytotoxicity for 3T3 fibroblast cells. In conclusion, the novel microwave-ultrasound hybridization technology allows the extraction of polyphenols that may have a potential cytotoxic effect on cancer cells.

Interaction of Bephratelloides cubensis (Hymenoptera: Eurytomidae) with Annona macroprophyllata fruits in orchards of Chiapas, Mexico / Interacción de Bephratelloides cubensis (Hymenoptera: Eurytomidae) con frutos de Annona macroprophyllata en huertos de Chiapas, México

Introduction: Annona macroprophyllata Donn. Smith. (Annonaceae) (syn. Annona diversifolia Saff.) is a valued fruit tree species known as papausa. In Mexico and Central America, this fruit has become an important crop because of its tasty flavor and high pulp content. Its fruits are frequently damaged by the incidence of wasps of the genus Bephratelloides Girault (Hymenoptera: Eurytomidae), which develop inside the seeds. Objective: To report the interaction of Bephratelloides cubensis Ashmead during its life cycle in fruits of A. macroprophyllata. Methods: We periodically collected fruits in different states of growth recording a) oviposition, b) the moment of evident infection, c) the development of the wasps inside the seeds, and d) their emergence as adults. We also determined the proportion of damaged fruits and seeds. Results: The data indicate that wasps preferred to oviposit on fruits with a diameter of less than 8 cm, oviposition was more frequent between 11:00 am and 03:00 pm., and there was 26 % infestation of fruits, and 9 % of seeds. Conclusion: It is an obligatory interaction for the wasp, the highest proportion of attack on fruits was in the early stages of fruit development and control actions should focus on the protection of these early stages.

Introducción: Annona macroprophyllata Donn. Smith. (Annonaceae) es una especie de árbol frutal, conocida como papausa. En México y América Central, esta fruta se ha convertido en un cultivo importante y valorado debido a su delicioso sabor y la cremosa textura de su pulpa. Sus frutos son frecuentemente dañados por la incidencia de avispas del género Bephratelloides Girault (Hymenoptera: Eurytomidae), que se desarrollan dentro de las semillas. Objetivo: Describir la interacción de Bephratelloides cubensis Ashmead durante su ciclo de vida en frutos de A. macroprophyllata. Método: Recolectamos frutos en diferentes estados de crecimiento registrando a) la ovoposición, b) el momento de la infección evidente, c) el desarrollo de las avispas dentro de las semillas, y d) su emergencia como adultos. También determinamos la proporción de frutas y semillas dañadas. Resultados: Los datos indican que las avispas preferían ovipositar en frutas con un diámetro de menos de 8 cm, la oviposición era más frecuente entre las 11:00 a.m. y las 3:00 p.m., y que había un 26 % de la infestación en las frutas y un 9 % en semillas. Conclusión: Es una interacción obligada para la avispa y la mayor proporción de ataque a los frutos fue en las etapas tempranas de desarrollo de frutos y las acciones de control se deberían enfocar a la protección de estas etapas.

Chemical characterization, antimicrobial and antioxidant activity of sugar-apple (Annona squamosa L. ) pulp extract / Caracterización química, actividad antimicrobiana y antioxidante del extracto de pulpa de manzana de azúcar (Annona squamosa L. )

Synthetic preservatives are widely present in processed foods, but most of them have carcinogenic potential, requiring the development of new natural alternatives such as fruit extracts, for microbial control. The objective of the study was to evaluate the chemical characterization, antioxidant, and antimicrobial activity of the sugar apple pulp (Annona squamosa L.). Physicochemical characteristics were evaluated, an extract was prepared, and its antioxidant activity by DPPH method and antimicrobial by disk diffusion. Minimal inhibitory concentration and minimum bactericidal concentration against strains of Salmonella typhimurium, Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus were evaluated. The physicochemical analysis revealed that sugar apple pulp had 75.0% moisture, 3.0% ash, 4.0% protein, 0.2% lipids, 3.3% fibers, and 14.5% carbohydrates. The antioxidant activity of the extract by the DPPH method was 20.6%. The pulp extract from the sugar apple had inhibition zone for Staphylococcus aureus, satisfactory inhibitory effect against Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, and Salmonella Typhimurium, but did not present a bactericidal effect. Sugar apple pulp presents adequate levels of nutrients and potential for food application due to its microbiological activity and antioxidant properties.

Los conservantes sintéticos están ampliamente presentes en los alimentos procesados, pero la mayoría tienen potencial carcinogénico, lo que requiere el desarrollo de nuevas alternativas naturales para el control microbiano, como los extractos de frutas. El objetivo del estudio fue evaluar la caracterización química, la actividad antioxidante y antimicrobiana de la pulpa de manzana de azúcar (Annona squamosa L.). Se evaluaron las características fisicoquímicas, y se evaluó su actividad antioxidante mediante el método DPPH y antimicrobiano por difusión en disco, concentración inhibitoria mínima y concentración bactericida mínima contra cepas de Salmonella Typhimurium, Escherichia coli, Listeria monocytogenes y Staphylococcus aureus. El análisis fisicoquímico reveló que la pulpa de manzana de azúcar tiene 75.0% de humedad, 3.0% de cenizas, 4.0% de proteínas, 0.2% de lípidos, 3.3% de fibras y 14.5% de carbohidratos. La actividad antioxidante del extracto por el método DPPH fue del 20.6%. El extracto de pulpa de la manzana de azúcar tenía zona de inhibición para Staphylococcus aureus, efecto inhibidor satisfactorio contra Staphylococcus aureus, Escherichia coli, Listeria monocytogenes y Salmonella Typhimurium, pero no presenta efecto bactericida. La pulpa de manzana de azúcar presenta niveles adecuados de nutrientes y potencial para la aplicación de alimentos debido a su actividad microbiológica y propiedades antioxidantes.

Estructura poblacional y hábitat de un árbol tropical con frutos comestibles, Annona purpurea (Annonaceae), en el occidente de México / Population structure and habitat of a tropical tree with edible fruits, Annona purpurea (Annonaceae), in Western Mexico

Introducción: Annona purpurea es un árbol Mesoamericano, distribuido por la vertiente pacífica y atlántica de México hasta Sudamérica. Diferentes partes de la planta son de utilidad y los frutos son comestibles. Objetivo: Describir la estructura poblacional, el hábitat y los factores ambientales que influyen en la distribución y abundancia de A. purpurea en el occidente de México. Métodos: De abril a noviembre 2015, se establecieron 24 unidades de muestreo de 500 m2 cada una, con presencia de A. purpurea. En cada sitio se midieron e identificaron todas las especies leñosas con diámetro a la altura de pecho (dap) ≥ 2.5 cm y se registró información ambiental, geográfica y climática. Se obtuvo información sobre la repoblación de la especie. Se estimó la estructura de A. purpurea y se examinaron sus relaciones con las variables ambientales. El índice de asociación de Whittaker se determinó utilizando la matriz de índice de valor de importancia (IVI). Con la matriz de variables ambientales y la de IVI, se realizó un Análisis de Correspondencia Canónica para determinar la influencia de las variables ambientales sobre la especie y visualizar la distribución de las especies en el espacio multidimensional. Resultados: A. purpurea presentó 1 108 tallos en 1.2 ha, con el 85 % de ellos concentrados en las tres primeras categorías diamétricas. Su densidad presentó una relación positiva con la presencia de tocones, mientras que el área basal y el IVI se relacionaron de manera similar con la incidencia de incendios. Hubo poca o nula regeneración de A. purpurea debajo de su dosel. Los mayores IVI en la comunidad se registraron para A. purpurea, Tabebuia rosea, Quercus magnoliifolia y Enterolobium cyclocarpum y la prueba de perfiles de semejanza separó a A. purpurea y Guazuma ulmifolia como un grupo diferente. Las variables ambientales con mayor influencia en la distribución y abundancia de A. purpurea fueron la precipitación anual, incidencia de incendios, elevación, temperatura media anual, pedregosidad y pastoreo. Conclusión: A. purpurea presenta los mayores atributos estructurales en la comunidad, sus poblaciones se ven favorecidas en sitios con mayor precipitación y temperatura, con poca pedregosidad, donde hay incidencia de perturbaciones por fuego y ganado que generan claros grandes y su mayor asociación la presenta con G. ulmifolia.

Introduction: Annona purpurea is a Mesoamerican tree, distributed from the Pacific and Atlantic slopes of Mexico to South America. Different parts of the plant are of utility and the fruits are edible. Objective: To describe the population structure, habitat and environmental factors that influence the distribution and abundance of A. purpurea in Western Mexico. Methods: From April to November 2015, 24 sampling units were established, each of which were 500 m2 in area and had a presence of A. purpurea. At each site, all woody species with a diameter at breast height (dap) ≥ 2.5 cm were measured and identified, and environmental, geographic and climatic information recorded. Information regarding the regeneration of the species was obtained. The structure of A. purpurea was estimated and its relationships with environmental variables examined. The Whittaker association index was determined using the importance value index matrix (IVI). With the matrix of environmental variables and that of IVI, a Canonical Correspondence Analysis was performed to determine the influence of environmental variables on the species and to visualize the distribution of the species in multidimensional space. Results: A. purpurea presented 1 108 stems in 1.2 ha, with 85 % of these concentrated in the first three diametric categories. Its density presented a positive relationship with the presence of stumps, while the basal area and IVI were similarly related to the incidence of fire. There was little or no regeneration of A. purpurea under its canopy. The highest IVI in the community were found for A. purpurea, Tabebuia rosea, Quercus magnoliifolia and Enterolobium cyclocarpum, and the similarity profiles test separated A. purpurea and Guazuma ulmifolia as a distinct group. The environmental variables with the greatest influence on the distribution and abundance of A. purpurea were annual precipitation, fire incidence, elevation, mean annual temperature, stoniness and livestock. Conclusion: A. purpurea presents the greatest structural attributes in the community, its populations are favored in places with greater precipitation and temperature, with little stoniness and with incidence of disturbances by fire and livestock that generate large gaps and its greatest association is with G. ulmifolia.

In vivo antileishmanial activity of Annona mucosa extracts

Abstract INTRODUCTION: Leishmaniasis, a disease caused by a parasite endemic to large areas of tropical and subtropical countries, is a growing public health problem. METHODS: Male BALB/c mice were infected with Leishmania amazonensis and treated with extracts isolated from Annona mucosa. RESULTS: Treated groups had significantly reduced footpad swelling. The group treated intraperitoneally with hexane extract showed footpad swelling similar to groups treated with Pentamidine® and Glucantime®. Groups treated with dichloromethane extract and hexane extract presented the recovering phenotype associated with reduced parasite levels. CONCLUSIONS: Extracts of A. mucosa are promising sources of novel antileishmanial compounds.

Efeito da concentração de polpa de graviola (Annona muricata L.) sobre a fermentação de mosto ácido de mel suplementado com extrato de feijão caupi (Vigna unguiculatana L. Walp.) para a produção de hidromel / Effect of graviola pulp concentration (Annona muricata L.) on fermentation of honey acid wort supplemented with caupi bean extract (Vigna unguiculatana L. Walp.) for the production of mead

O hidromel é uma bebida oriunda da fermentação alcoólica de mosto de mel e alguns adjuntos, como as frutas, para o fornecimento de nutrientes essenciais à fermentação. A graviola e o feijão caupi são importantes matérias-primas vegetais brasileiras, que constituem importantes fontes de carboidratos, proteínas, sais minerais e outros compostos. Assim, o presente trabalho teve como objetivo avaliar o efeito da suplementação de mostos de mel, com concentração fixa de feijão caupi (30 g.L-1) e diferentes concentrações de graviola (0, 10 e 30% m/m), sobre a fermentação dos mesmos, avaliando os perfis de concentração celular e dos teores de sólidos solúveis totais e etanol. Diante dos resultados obtidos foi possível observar que a graviola pode ser considerada um bom suplemento para o estímulo da fermentação de mostos de mel por leveduras.

Avaliação sensorial e intenção de compra de doce em massa de graviola / Sensory evaluation and intention to purchase of candy in graviola mass

Objetivou-se desenvolver e avaliar sensorialmente doces em massa de graviola com substituição parcial da sacarose por xarope de glicose. As formulações de doces foram elaboradas variando a concentração de xarope de glicose em substituição a sacarose. A análise sensorial foi aprovada pelo comitê de ética em pesquisa da Universidade Estadual da Paraíba. Os doces apresentaram cor, aroma e sabor característicos do fruto. Percebeu-se que a aceitação sensorial apresentou semelhança significativas entre as formulações e boa impressão global, ficando com média que remete a “gostei moderadamente”. Todas as formulações alcançaram valores satisfatórios para a avaliação sensorial, sendo a formulação com substituição parcial de 19,70% a que apresentou maior aceitação.

Eight new cytotoxic annonaceous acetogenins from the seeds of Annona squamosa / 中国天然药物

Eight new annonaceous acetogenins, squamotin A-D (1-4), annosquatin IV-V (5 and 6), muricin O (7) and squamosten B (8), together with four known ones (9-12) were isolated from the seeds of Annona squamosa. Their structures were elucidated by chemical methods and spectral data. The inhibitory activities of compound 1-9 against three multidrug resistance cell lines were evaluated. All tested compounds showed strong cytotoxicity.

Formation of soursop seedlings irrigated using waters with different salinity levels and nitrogen fertilization / Formação de mudas de gravioleira irrigadas com águas de distintas salinidades e adubação nitrogenada

Using saline waters in agriculture has become common in many regions worldwide, but some techniques have been developed to enable the use of these waters in order not to harm the crop. Thus, this study aimed to evaluate the growth of soursop seedlings, cv. 'Morada Nova', under interaction between salt stress and nitrogen (N) fertilization. The experiment was conducted in a greenhouse in randomized block design, in 5 x 4 factorial scheme, with 4 replicates, formed by the combination of five levels of irrigation water electrical conductivity - ECw (0.3; 1.1; 1.9; 2.4 and 3.5 dS m-¹) and four N doses (70, 100, 130 and 160%). The dose relative to 100% corresponded to 100 mg of N dm-³ of soil. The interaction between N doses and water salinity levels did not affect the seedling production stage of soursop, cv. 'Morada Nova'. The growth of 'Morada Nova' soursop seedlings subjected to different water salinity levels was less affected in the initial stage (45 days after treatment application). Water with ECw of 2.0 dS m-¹ can be used to produce soursop seedlings, because it leads to an acceptable mean growth reduction of 10%. N doses higher than 70 mg dm-³ do not either attenuate salt stress or promote higher growth of soursop seedlings, cv. 'Morada Nova'.

O uso de águas salinas na agricultura tornou-se um fato corriqueiro em diversas regiões do mundo. Contudo algumas técnicas têm sido desenvolvidas para viabilizar o uso dessas águas de modo a não prejudicar a cultura. Neste sentido, objetivou-se avaliar o crescimento de mudas de gravioleira cv. Morada Nova sob interação entre a salinidade da água de irrigação e adubação nitrogenada. Desenvolvida em casa de vegetação em delineamento de blocos casualizados em esquema fatorial 5 x 4, com quatro repetições, constituído pela combinação de cinco condutividade elétrica da água de irrigação CEa (0.3; 1.1; 1.9; 2.4 e 3.5 dS m-¹) e quatro doses (70, 100, 130 e 160% de nitrogênio). Sendo a dose referente a 100%, correspondente a 100 mg de N por dm-³ de solo. A interação entre os fatores doses de nitrogênio e níveis de salinidade da água não afetaram a fase de produção de mudas de gravioleira cv. Morada Nova. O crescimento das mudas de gravioleira cv. Morada Nova, submetidas a diferentes níveis de salinidade da água foi menos comprometido pela salinidade na fase inicial (45 Dias após aplicação dos tratamentos). Na produção de mudas de gravioleira pode-se usar água de CEa de até 2.0 dS m-¹ pois proporciona redução média aceitável de 10% no crescimento. Doses de nitrogênio superior a 70 mg de N dm-³ de solo não atenuam o estresse salino nem promovem maior crescimento de mudas de gravioleira cv. Morada Nova.

Physical-chemical characterization of powder soursop obtained by foam-mat drying / Caracterização físico-química de pó de graviola obtido por secagem em leito de espuma

The development and improvement of fruit processing technologies in powder form is a alternative to add value to the product and increase the income of the producers, being required the knowledge of physicalchemical properties of the product. The aim of this study was to determine the physical-chemical properties of soursop powder obtained by foam-mat drying. To the foam formation, albumin was added to the pulp at concentration of 7.43% in mass and subjected in mixer for 15 min; then, it was spread onto trays forming a thin layer about 5.0 mm thick, and the drying conditions were: 40, 50, 60, 70 and 80 °C, 5.6 m s-1 and 60%. Moisture content, water activity, titratable total acidity and hydrogenic potential of soursop powder indicate good stability of the product obtained by foam-mat drying, without the need of addition of preservatives for its conservation. Reducing sugars increased with increasing drying temperature due to the concentration of the compounds caused by moisture content reduction. In addition, the color was the physical-chemical property that presented greater sensitivity to drying, mainly by darkening index. Temperatures lower than 60 °C indicate higher preservation of initial characteristics for soursop powder. Besides that, bulk density, true density, porosity and repose angle increased subtly with drying temperature.

O desenvolvimento e aperfeiçoamento de tecnologias de processamentos de frutos na forma de pó é uma alternativa para agregar valor ao produto e aumentar a renda dos produtores, sendo necessário para isso o conhecimento das propriedades físico-químicas do produto. Assim objetivou-se determinar as propriedades físicoquímicas do pó de graviola obtido pelo método de secagem em leito de espuma. Para a formação da espuma foi adicionada, à polpa, albumina, na concentração de 7,43% em massa e submetida à agitação em batedeira doméstica, durante 15 min; em seguida, esta foi espalhada sobre bandejas formando uma camada fina de cerca de 5,0 mm de espessura cujas condições de secagem foram: de 40, 50, 60, 70 e 80 °C, 5,6 m s-1 e 60%. O teor de água, atividade de água, acidez total titulável e o potencial hidrogeniônico do pó de graviola indicam boa estabilidade do produto obtido pelo método de secagem em leito de espuma, não necessitando portanto, da adição de conservantes para sua conservação. Os açúcares aumentaram com o incremento da temperatura de secagem devido a concentração dos compostos provocada pela redução do teor de água. Adicionalmente, a cor foi a propriedade físico-química que apresentou maior sensibilidade à secagem, principalmente pelo índice de escurecimento. Temperaturas menores que 60 °C indicam maior preservação das características iniciais para o pó de graviola. Além disso, a massa específica aparente e unitária, a porosidade e o ângulo de repouso aumentaram sutilmente com a temperatura de secagem.

Population fluctuations in the pink hibiscus mealybug and its natural enemies in Annona squamosa (Annonaceae) in Roraima, Brazil / Flutuação populacional da cochonilha-rosada e seus inimigos naturais em Annona squamosa (Annonaceae) em Roraima, Brasil

Maconellicoccus hirsutus (Hemiptera, Pseudococcidae), a species of economic interest, especially for fruit plants, is expanding on the South American continent. Information about the population dynamics of this pest associated with control by natural enemies and cultural practices is fundamental for its management. Our objective was to study the population fluctuations in M. hirsutus and its natural enemies in a sugar-apple (Annona squamosa) orchard in Roraima, northern Brazil. Trees were evaluated monthly over a 12-month period. Infestation rates by M. hirsutus and its parasitism were also estimated for potential host plants around the study area. Highest infestation occurred in August and February-March. Alternative hosts were infested during the off-season, mainly fruit. Lacewings and the parasitoid Anagyrus kamali (Hymenoptera, Encyrtidae) were abundant natural enemies. Average parasitism by A. kamali in fruits was 50%, with highest rates in periods of greatest infestation by M. hirsutus. Fruitification pruning reduced M. hirsutus populations.

Maconellicoccus hirsutus (Hemiptera, Pseudococcidae), praga de inúmeras espécies, especialmente frutíferas, está se expandindo no continente sul-americano. Informações sobre sua dinâmica, inimigos naturais e práticas culturais são importantes para seu manejo. Nosso objetivo foi estudar as flutuações da população em M. hirsutus e seus inimigos naturais em um pomar de ata (Annona squamosa) em Roraima, Brasil. As árvores foram avaliadas mensalmente ao longo de um período de 12 meses e as taxas de infestação por M. hirsutus e parasitismo foram estimadas. A maior infestação ocorreu em agosto de 2014 e entre fevereiro e março de 2015. Hospedeiros alternativos foram infestados durante a entressafra da ata. Bicho-lixeiro e o parasitoide Anagyrus kamali (Hymenoptera, Encyrtidae) foram os inimigos naturais mais importantes. O parasitismo médio por A. kamali nos frutos foi de 50%, com maiores taxas nos períodos de maior infestação por M. hirsutus. A poda de frutificação reduziu as populações de M. hirsutus.