In silico screening of Annona Muricata L. Leaf constituents and derivatives towards selected targets of Type 2 Diabetes Mellitus

BACKGROUND@#Type 2 diabetes mellitus, or T2DM, is one of the world's most chronic health problems that is linked to numerous deaths and high health care expenses. 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), protein-tyrosine phosphatase 1B (PTP1B) and mono-ADP-ribosyl transferase sirtuin-6 (SIRT6) were among the novel proteins and focus targets of diabetes research. Annona muricata is a commonly used natural remedy for several illnesses, including type 2 diabetes mellitus. However, most of these traditional claims have received few molecular evaluations.@*OBJECTIVES@#This investigated the phytoconstituents and derivatives of the leaves of A. muricata by evaluating their binding profiles towards selected novel T2DM-related protein targets through in silico methods.@*METHODOLOGY@#This study screened the potential lead compounds from the leaves of A. muricata by evaluating the binding energies of the parent compounds and derivatives with the targets compared to the native ligands and known substrates through molecular docking simulations. Additionally, pharmacokinetic, physicochemical properties, and binding interactions were also assessed using several software programs and online databases.@*RESULTS@#Out of the 8 selected parent compounds of Annona muricata, a total of 672 derivatives were designed, tested, and compared against the controls for at least one of the three protein targets. Among these, 280 derivatives exhibited more negative binding energies than controls in each protein target.@*CONCLUSION@#The designed derivatives can be synthesized and further investigated for potential biological effects towards 11β-HSD1, PTP1B, and SIRT6 through in vitro and in vivo experiments.

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

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.

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.

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.

Annona muricata Linn Stem Bark Protects against Uterine Proliferative Disorder Induced by Estradiol Benzoate in Female Rat

Aim: The opening of mitochondrial permeability transition (mPT) pore is an important event in the execution of mitochondrial-mediated apoptosis. Some bioactive compounds elicit their chemotherapeutic effects against tumor/cancer cells via the induction of mitochondrial-mediated apoptosis. Annona muricata, a medicinal plant, is folklorically used in the treatment of tumors and cancers. This study therefore aimed at investigating the effect of methanol stem bark extract of Annona muricata (MEAM) on apoptosis via mPT pore and estradiol benzoate (EB)-induced proliferative disorder using female Wistar rats. Methodology: Mitochondria were isolated using differential centrifugation. The mPT pore opening, cytochrome c release and mitochondrial ATPase activity were determined spectrophotometrically. The levels of estrogen (E2), luteinizing hormone (LH), follicle stimulating hormone (FSH), malondialdehyde (MDA) and activities of superoxide dismutase (SOD), glutathione peroxidase (GSH), were determined using ELISA technique. Histological and histochemical assessments of the uterine sections were carried out using standard methods. Phytochemical constituents of MEAM were determined using Gas Chromatography-Mass Spectroscopy (GC-MS). Results: The in vitro results showed a significant induction of mPT pore opening, release of cytochrome c and enhancement of mitochondrial ATPase (mATPase) activity in a concentration-dependent manner. However, oral administration of MEAM did not induce rat uterine mPT pore opening, neither any significant release of cytochrome c nor enhancement of mATPase activity at the dosages used. Interestingly, MEAM reversed the EB-induced increase in E2, LH and FSH. The MEAM also improved the antioxidant milleu by reducing MDA level and increasing the SOD and GSH-Px activities in the treatment groups. Administration of EB induced endometrial hyperplasia in the model group which was mitigated by MEAM in the treatment group. The GC-MS analysis of MEAM revealed the presence of some important phytochemicals that are pharmacological relevant in cancer treatment. Conclusions: This study suggests that the methanol stem bark extract of Annona muricata contains bioactive compounds that protect against EB-induced uterine proliferative disorder in female Wistar rats.

Soursop (Annona muricata L.) Fruit Peels as Source of Phenolic Constituents and Annonacin with Biological Activities

Aims: In this study, chemical constituents and biological activities of the Annona muricata L. fruit peels were evaluated using methanol extract (MEAM) and hexane (HFAM), dichloromethane (DFAM), ethyl acetate (EFAM), and butanol (BFAM) fractions. Place and Duration of Study: All the experiments were done in the Department of Pharmaceutical Sciences and Department of Biochemistry, Federal University of Juiz de Fora, Juiz de Fora, Minas Gerais, 36026-900, Brazil, between January 2012 and July 2016. Methodology: Phytochemical screening (specific chemical reactions), total phenolic and flavonoid contents (Spectrophotometric methods) and chemical compounds were assessed (High performance liquid chromatography analysis). The antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), beta-carotene, and thiobarbituric acid assays. The inhibitory effect against digestive enzymes (lipase, ?-amylase and ?-glucosidase) was measured by spectrophotometric assays and and toxicity by the brine shrimp lethality bioassay. Results: Tannins, flavonoids, coumarins, terpenes and steroids, saponins, and alkaloids were detected. EFAM had the highest values of total phenolic and flavonoids, while a similar compound to annonacin was found in MEAM by HPLC. EFAM was also more active in DPPH and FRAP assays, and HFAM was effective in inhibiting the linoleic acid oxidation and the malondialdehyde. MEAM and fractions blocked lipase, ?-amylase and ?-glucosidase, while HFAM and DFAM were toxic against Artemia salina. Conclusion: The results showed that the A. muricata fruit peels have important biological effects, which can bring great benefits to human and animal health.

Phytochemical Screening of Aqueous and ethanolic, stem extract of Annona squamosa Linn.

Annona squamosa is an evergreen shrub belonging to the family Annonaceae. These plants have medicinal importance, widely used in preparation of anti head like medicines. It has anticancerous activity, used as antioxidants. The Phytochemical present in these plants also shows the antimicrobial, larvicidal and insecticidal activity. Plant chemical also inhibit the growth of pathogenic bacteria and fungi. In present study, we focused on the Phytochemicals in stem bark of A. squamosa plant. For their extraction simple soxhlet extraction (Clevenger apparatus) was carried out by using different solvent that is aqueous and ethanol. In qualitative analysis alkaloids, proteins, carbohydrates, steroids, tannins, oxalate, quinine, phenols, amino acid were found in ethanolic extract except saponin and flavonoid while in aqueous extract all Phytochemicals were present but phenol and tannins are absent.

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.

Cytotoxic effect of hydroalcoholic extract of Annona Muricata against a human cell line of gastric adenocarcinoma / Efecto citotóxico del extracto hidroalcoholico de la Annona Muricata frente a la línea celular humana de adenocarcinoma gástrico

Background: Therapeutic advances against cancer have not been as successful as expected and have adverse effects that patients rarely tolerate. A study in Peru identified favorable anticancer effects of Annona muricata (AM), a medicinal plant known as soursop, in C-678 mouse gastric adenocarcinoma. However, to date, no results have been reported in human cells. Objective: The objective of this study was to determine the cytotoxic effect of AM extract against a human gastric adenocarcinoma cell line (AGS). Methodology: Experimental in vitro analytical study using a hydroalcoholic extract of AM (AMOH) leaves collected in the Amazonas. Chemical functional groups were identified by phytochemical screening. To obtain the cytotoxic effect, different dilutions of extract were added to the plates containing the cell lines and the data were extrapolated to GraphPad employing an observation card. Finally, the cytotoxic effect was expressed as the half-maximal inhibitory concentration (IC50) and nonlinear regression analysis was performed to determine the growth inhibition of cancer cells. Results: Phytochemical screening showed the presence of reducing carbohydrates, alkaloids, phenols, tannins, triterpenes, steroids, saponins, flavonoids, proteins, cardiac glycosides, and anthocyanins. A calibration curve with gallic acid was used to determine the total phenol content and, quercetin was used to identify the flavonoid content. The AGS cell line showed cytotoxic activity with AMOH with an IC50 at 24 hours of 45.81 µg/mL and 19.05 µg/mL at 48 hours. Conclusion: Several chemical functional groups of AM were identified. In addition, the AMOH showed a cytotoxic effect against the AGS cell line

Antecedente: Los avances terapéuticos frente al cáncer no han tenido el éxito esperado y presentan efectos adversos pocas veces tolerados por el paciente. Un estudio en Perú identificó el efecto anticancerígeno de la Annona muricata (AM), planta medicinal conocida como guanábana, en adenocarcinoma gástrico de ratón C-678 con resultados favorables, sin embargo, no se ha encontrado evidencia previa en células humanas. Objetivo: El objetivo de este estudio fue determinar el efecto citotóxico del extracto de AM frente a la línea celular de adenocarcinoma gástrico humano (AGS). Materiales y métodos: Estudio experimental in vitro tipo analítico con extracto hidroalcohólico de hojas de AM (AMOH) recolectadas en Amazonas. Mediante screening fitoquímico se identificaron los grupos funcionales químicos. Para obtener el efecto citotóxico, se añadieron diferentes diluciones de extracto a las placas que contienen las líneas celulares y mediante una ficha de observación los datos fueron extrapolados a GraphPad. Finalmente se expresó como la concentración inhibitoria media máxima (IC50) y se hizo un análisis de regresión no lineal con la finalidad de encontrar la cantidad de inhibición de crecimiento de células oncológicas. Resultados: En el screening fitoquímico se pudo identificar la presencia de carbohidratos reductores, alcaloides, fenoles, taninos, tritérpenos y esteroides, saponinas, flavonoides, proteínas, glicósidos cardiotónicos y antocianinas. Para identificar el contenido total de fenoles se utilizó la curva de calibración con ácido gálico el cual nos comprobó la presencia de una buena cantidad de estos metabolitos. Adicionalmente se utilizó quercetina para identificar el contenido de flavonoides, obteniendo resultados favorables ya que se hizo evidente su presencia. La línea celular AGS mostró una actividad citotóxica frente al AMOH con un IC50 a las 24 horas de 45.81ug/mL y 19.05ug/mL a las 48 horas. Conclusión: Se identifica a los grupos funcionales de la AM. Además, AMOH demostró un efecto citotóxico contra la línea celular AGS

Control de larvas de Aedes aegypti con extractos de Allium sativum y Annona muricata como larvicidas / Control of larvae Aedes aegypti with extracts of Allium sativum and Annona muricata as larvicides

RESUMEN Objetivo Evaluar el efecto larvicida en éter de petróleo de los extractos de Allium sativum (ajo) y Annona muricata (guanábana) sobre larvas en IV estadio de Aedes aegypti en condiciones de laboratorio. Métodos Se realizaron diferentes bioensayos (tratamientos) en 6 concentraciones para Annona muricata y 7 concentraciones de Allium sativum, con cuatro repeticiones y un control. Se tuvo lecturas de mortalidad a las 2, 12, 24, 36 y 48 horas. Se validaron los datos obtenidos estadísticamente (corrección de Abbott y Análisis ANOVA). Además, se determinaron las concentraciones y tiempos letales para ambos extractos con un análisis Probit. Resultados Se obtuvo que, en un periodo de 48 horas, el tratamiento de 500 ppm del extracto de Annona muricata logró una mortalidad del 97%, mientras que el tratamiento de 2000 ppm con Allium sativum logró alcanzar una mortalidad del 85%. El tiempo letal 50 (50% de mortalidad) para Annona muricata, se obtuvo en el tratamiento de 200 ppm antes de 24 horas, para el caso de Allium sativum fue en el tratamiento de 1200 ppm antes de 48 horas. Para el tiempo letal 90 (90% de mortalidad) para Annona muricata, se obtuvo en el tratamiento de 400 ppm antes de 40 horas. Para el caso de Allium sativum, el tiempo letal 90 no fue posible obtenerlo experimentalmente. Se determinó por medio de un modelo matemático lineal, que dio como resultado 51 horas. Conclusión Ambas especies poseen efecto larvicida. Sin embargo, el extracto más eficiente y efectivo como larvicida es el de Annona muricata, lo que permite dar una alternativa natural, viable, económica y biodegradable para el control de larvas de esta especie.

ABSTRACT Objective To evaluate the larvicidal effect in petroleum ether of the extracts of Allium sativum (garlic) and Annona muricata (soursop) on larvae in stage IV of Aedes aegypti under laboratory conditions. Methods Different bioassays (treatments) were performed in 6 factors for Annona muricata and 7 concentrations of Allium sativum, with four replications and one control. Mortality readings were taken at 2, 12, 24, 36 and 48 hours. The data obtained statistically (Abbott correction and ANOVA analysis) were validated, in addition, the concentrations and lethal times for both extracts were determined with a Probit analysis. Results It was obtained that, in a 48 hour period, the treatment of 500 ppm of the extract of Annona muricata resulted in a mortality of 97%, while the treatment of 2000 ppm with Allium sativum reached a mortality of 85%. The lethal time 50 (50% mortality) for Annona muricata, was obtained in the treatment of 200 ppm within 24 hours, in the case of Allium sativum it was in the treatment of 1200 ppm before 48 hours. For the lethal time 90 for Annona muricata, obtain the treatment of 400 ppm before 40 hours, for the case of Allium sativum, the lethal time 90 (90% mortality) could not be obtained experimentally, it was determined by means of a linear mathematical model, resulting in 51 hours. Conclusion Both species affected larvicidal effect. However, the most efficient and effective extract as a larvicide is that of Annona muricata, which allows giving a natural, viable, economical and biodegradable alternative for the control of larvae of this species.

Efecto antibacteriano del extracto etanólico de Annona muricata sobre microorganismos de importancia clínica / Antibacterial effects of Annona muricata ethanolic extract on clinically important microorganisms

El incremento de cepas patógenas resistentes a fármacos convencionales ha limitado las opciones de tratamiento médico. Ante ello surge la necesidad de buscar alternativas terapéuticas. Muchas especies vegetales poseen enorme potencial antimicrobiano que puede ser de gran utilidad. Objetivo: determinar el efecto antibacteriano in vitro del extracto etanólico de Annona muricata L. sobre Staphylococcus aureus, Streptococcus B - hemolíticos y Escherichia coli. Métodos: se evaluaron 135 unidades experimentales conformadas por 3 cepas de Staphylococcus aureus, Streptococcus B-hemolíticos y Escherichia coli, además de 5 concentraciones del extracto y 3 repeticiones del experimento. Para determinar el efecto antibacteriano in vitro se emplearon los métodos de disco difusión en agar y macrodilución en caldo. Se utilizó el extracto etanólico a concentraciones de 125, 250, 500, 750, 1000 mg/ml y solución salina fisiológica estéril como control negativo. Resultados: el extracto inhibió el crecimiento in vitro de Staphylococcus aureus y Streptococcus B-hemolíticos. La mayor inhibición se observó a 1 000 mg/ml con halos inhibitorios de 14,6 mm y 12,33 mm de diámetro, respectivamente. Para Escherichia coli no se observó la formación de halos inhibitorios. Las cepas de Streptococcus B-hemolíticos y Staphylococcus aureus presentaron una concentración mínima inhibitoria de 250 y 500 mg/ml, respectivamente. Conclusión: el efecto antibacteriano in vitro fue directamente proporcional a cada concentración empleada sobre Staphylococcus aureus y Streptococcus B-hemolíticos. En el caso Escherichia coli no se observó inhibición de crecimiento.

The increase in pathogenic strains resistant to conventional drugs has limited medical treatment options. Given this, the need to seek therapeutic alternatives arises. Many plant species have enormous antimicrobial potential that can be very useful. Objective: to determine in vitro the antibacterial effect of the ethanolic extract of Annona muricata L. on Staphylococcus aureus, Streptococcus B - hemolytic and Escherichia coli. Methods: 135 experimental units were evaluated, consisting of 3 strains of Staphylococcus aureus, Streptococcus B-hemolytic and Escherichia coli, in addition to 5 concentrations of the extract and 3 repetitions of the experiment. To determine the antibacterial effect in vitro, the agar diffusion disk and broth macrodilution methods were used. The ethanolic extract was used at concentrations of 125, 250, 500, 750, 1000 mg / ml and sterile physiological saline solution as negative control. Results: the extract inhibited in vitro growth of Staphylococcus aureus and Streptococcus B-hemolytic. The greatest inhibition was observed at 1000 mg / ml with inhibitory halos of 14.6 mm and 12.33 mm in diameter, respectively. For Escherichia coli the formation of inhibitory halos was not observed. The Streptococcus B-hemolytic and Staphylococcus aureus strains presented a minimum inhibitory concentration of 250 and 500 mg / ml, respectively. Conclusion: the in vitro antibacterial effect was directly proportional to each concentration used on the Staphylococcus aureus and Streptococcus B-hemolytic. In the case of Escherichia coli, no growth inhibition was observed.

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

Development, Characterization and Microbiological Evaluation of Silver Nanoparticles, Obtained by Green Synthesis, from the Extract of Annona muricata L

Abstract Green synthesis is an efficient method, frequently applied in nanobiotechnology, as it does not use toxic reagents or solvents. Biological organisms, including medicinal plants, have been used successfully for manufacturing of different types of metallic nanoparticles. The aim of this work was to synthesize, characterize and evaluate the antimicrobial activity of silver nanoparticles (AgNPs) obtained from the extract of the pulp, seed and leaves of Annona muricata L. The particle size, polydispersity index, zeta potential, as well as morphological aspects of AgNPs were evaluated. With the data obtained from the analyses, we concluded that the nanoparticles were successfully obtained by a simple and green method using the aqueous extract of the pulp, seeds and leaves of A. muricata. AgNPs obtained by A. muricata pulp extract without exposure to artificial light showed lowest average of hydrodynamic diameter and smallest size at field emission scanning electron microscope (FESEM). In addition, these nanoparticles showed the best polydispersity index (PDI), and zeta potential of - 29.6, which indicates good stability. AgNPs obtained from the extract of the pulp, seed and leaves showed antimicrobial activity, against strains of gram positive and gram negative bacteria, and antifungal activity, compared to the pure extract.

Potential Role Of Annona Squamosa Linn Extract And N-Acetyl Cysteine In Management Of Rifampicin-Induced Hepatoxicity In Rats

Objective: Rifampicin (RIF) could be a recognized therapeutic and preventive agent against tuberculosis. Still, high rates of many side effects and symptoms related to hepatotoxicity have identified during treatment. So, the current study was aimed to evaluate the antioxidant and hepatoprotective activity of methanolic extract of Annona Squamosa Linn (MEAS) and N-Acetyl Cysteine (NAC) against RIF induced hepatic injury in male rats. Methods: The hepatoprotective effects of MEAS (500 mg/kg b.wt.) and NAC (100 mg/kg b.wt.) or co-treatment were assessed in a model of hepatotoxicity by RIF (300 mg/kg b. wt.) in male rats daily for 21 d. Moreover, bilirubin, total protein, albumin, ALT, AST, ALP, GGT, MDA, and GSH were estimated. In addition, the levels of IL-6, IL-10, 8(OH)dG, and Bcl2 were evaluated. Results: The oral administration of MEAS and NAC or their combination resulted in significant reductions in the levels of bilirubin, albumin, hepato-specific markers namely ALT, AST, ALP, GGT, and MDA as compared to the RIF group. Furthermore, MEAS and NAC or the combination of MEAS and NAC treatment significantly up-regulated the levels of total protein, glutathione reductase with concomitant decrease in inflammatory marker level IL-6 and apoptotic marker level 8(OH)dG as well as increment the level of anti-inflammatory marker IL-10 and anti-apoptotic marker Bcl2 as compared to the RIF group. Histological examination of the liver tissue indicated that co-treatment with MEAS and NAC completely abolished the inflammation and degeneration in hepatocytes and restore the liver tissue to its normal structure. Conclusion: The present findings demonstrated that a co-treatment of MEAS and NAC seems to be more productive and curative than alone MEAS or NAC treatment and strongly compensated the liver damage induced by RIF.

Antioxidant Activity And Total Phenol Content Of Different Plant Parts Of Lebanese Annona Squamosa Linn.

Objective: The aim of this work was to characterize the antioxidant properties and to evaluate the total phenol content of leaves, bark, pericarp, and pulp extracts of Lebanese Annona squamosa Linn. (A. squamosa),, as well as a total screening of secondary metabolites present in the various plant parts studied. Methods: Two solvent systems were used for extraction: ethanol 80 % and methanol 80 %. The antioxidant activity of different extracts was investigated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The Total Phenol Content (TPC) of the different plant parts are determined and compared via Folin-Ciocalteu method. The results were presented as the mean of three separate experiments and error bars were used to illustrate standard deviation. Results: The phenolic content was found to be highest in the A. squamosa leaves methanolic and ethanolic extracts (117.2 mg and 112.92 gallic acid extract/g, respectively). The results showed that A. squamosa leaves methanolic and ethanolic extracts display the highest antioxidant activities than the bark, pulp and pericarp extracts, with half-maximal inhibitory concentration values 13.61 and 15.97 μg. ml-1 respectively. Ethanol 80 % and methanol 80 % were found to be efficient for the extraction of phenolic compounds. Conclusion: Results of this study indicate the presence of promising compounds in Lebanese A. squamosa that are able to act as antioxidants and free radical scavengers.

RECK and TIMP-2 mediate inhibition of MMP-2 and MMP-9 by Annona muricata

Up-regulation of MMP-2 and MMP-9 plays a significant role in promoting cancer progression by degradingthe components of the extracellular matrix, thereby enhancing the migration of tumor cells. Although the antiproliferative and apoptotic effect of Annona muricata is well established, its effect on MMP-2 and MMP-9, amajor target in several types of cancers, has not been studied. Powdered samples of various parts of A.muricata like fruit, stem, seed, and twig extracted using aqueous methanol showed significant dose-dependentinhibition of MMP-2 and MMP-9 in a highly metastatic fibrosarcoma cell line, HT1080. Additionally, theseextracts also up-regulated the expression of several endogenous inhibitors of MMP-2 and MMP-9 likeREversion-inducing Cysteine-rich protein with Kazal motifs (RECK) and Tissue Inhibitor of Metalloproteinase-2 (TIMP-2). Furthermore, primary cells developed from tumor tissues obtained from patients notexposed to chemotherapy, also exhibited similar results. Remarkably, the inhibition of MMP-2 and MMP-9observed was tumor specific, with the A. muricata fruit extract showing only 2% inhibition in cells obtainedfrom normal tissues, when compared to 60% inhibition observed in cells obtained from tumor samples. Thepresent study elucidates a novel mechanism by which A. muricata extracts selectively exhibit their anti-canceractivity in tumor cells by down-regulating MMP-2 and MMP-9 that are important biomarkers in cancer.

In Vitro Antidiabetic Activity By Glucose Uptake Of Yeast Cell Assay And Antioxidant Potential Of Annona Reticulata L. Leaf Extracts

The undesirable adverse effects of present available synthetic drugs endorse the modern medicine to search the superior choice for the treatment of metabolic diseases. Herbal medicine turns out to be a hopeful therapy for the effective treatment of diabetes in foreseeable future. In an effort to render a scientific evidence for the antidiabetic potential of Annona reticulata L., the present research, with the objective to evaluate the ability of A. reticulata leaf extracts on antihyperglycemic property under in vitro using yeast cell model was performed. Besides, as the diabetes and its complications are highly associated with the oxidative stress, the current study was also focussed on antioxidant property of A. reticulata leaf extracts. The scavenging ability of plant extracts on free radical 2, 2 – diphenyl- 1- picrylhydrazyl (DPPH), ferric reducing power assay and total antioxidant activity were performed to establish the antioxidant potential of A. reticulata leaf extracts. The in vitro antidiabetic ability of A. reticulata leaves was evaluated by glucose uptake method using yeast cell model. Among the four chosen solvent extracts- aqueous, methanol, ethyl acetate and n-hexane, methanol extract (MeE) exhibited high antioxidant potential. MeE exhibited 62.28% of DPPH inhibition at 200µg/ml, with total antioxidant activity 164.72 ± 2.63µg/ml and higher absorbance in reducing power assay (1.15 ± 0.03). In vitro antidiabetic activity by glucose uptake of yeast cell assay was evaluated and observed dose dependant rise in % of glucose uptake in methanol, ethylacetate and aqueous extracts of A. reticulata. MeE showed 48.55% of glucose uptake at 500µg/ml concentration. Hence the present study could be concluded as A.reticulata leaves possess potent antioxidant activity and antidiabetic activity under in vitro conditions. With the outcome of the present initial study, research work could be extended further; thereby the exact pharmacological action of the plant compounds could be discovered.

Nutrient and Bioactive Components of Annona muricata and Fagara zanthoxyloide of South-Southern Nigeria

Introduction:Prediabetes is associated with dysglycemia, endothelial dysfunction,obesity and inflammation, placing them at an increased risk of cardiovascular events.Aims: The present study aimed to investigate the risk of cardiovascular disease associated with prediabetes by estimation of serum interleukin-6, myeloperoxidase and urine microalbumin and their correlation with fasting plasma glucose and anthropometric measurements.Study Design:Cross sectional study.Place and Duration of Study: Study was conducted at Department of Biochemistry, Kasturba Medical College Hospitals, Mangaluru between 2014 and 2015.Methodology:Eighty subjects were categorised into prediabetes and healthy controls based on their fasting plasma glucose values. Anthropometric data (weight, body mass index, waist circumference, hip circumference and waist-to-hip ratio from all subjects were recorded. Interleukin-6 & myeloperoxidase were estimated in serum sample whereas microalbumin was estimated in random urine sample Results:The mean anthropometric measurements and cardiovascular disease risk markers (interleukin-6, myeloperoxidase and urine microalbumin) were found to be significantly higher (p < 0.05) in prediabetes group. Myeloperoxidase had significant correlation with fasting plasma glucose (r-0.388) in the prediabetes group. Interleukin-6 and myeloperoxidase also showed a positive correlation with body mass index (r -0.339, r -0.327), waist circumference (r -484, r -0.493) and waist-to-hip ratio (r -0.430, r -0.493) while urine microalbumin did not correlate with fasting plasma glucose and anthropometric measurements in prediabetes group.Conclusion:This study suggests that prediabetes is associated with central adiposity and have an increased risk for cardiovascular disease.

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.