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.

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.

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.

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.

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.

Niosomes and liposomes as promising carriers for dermal delivery of Annona squamosa extract

The leaf extract of Annona squamosa L. has antibacterial, antidiabetic, antioxidant, and anticancerous activities. The present work aims to compare between liposomes and niosomes as carriers for A. squamosa extract to improve its transdermal bioavailability. Physical characterization for niosomes and liposomes was performed using: transmission electron microscope (TEM), scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR). In addition, the encapsulation efficiency for A. squamosa in both carriers was evaluated and in-vitro drug release experiments were performed. The results proved the potential of both carriers to penetrate the outer layer of the skin (stratum corneum) which is considered as a strong barrier against the diffusion of many compounds through the skin. Moreover, the results pointed out that niosomes and liposomes lasted long time through the skin, which ensures the presence of antioxidant extract in the skin for prolonged periods. This would have a benefit of targeting free radicals in the skin. The encapsulation efficiency of liposomes for A. squamosa extract exceeded that of niosomes, however, niosomes demonstrated longer time of drug release through the skin. In conclusion, niosomes and liposomes are promising carriers for dermal delivery of the antioxidant extract Annona squamosa.

Gibberellic acid combined with hand pollination increases 'Red' and 'Lessard Thai' sugar apple fruit quality and produced parthenocarpic 'Gefner' atemoya fruits / Ácido giberélico associado com a polinização manual incrementa a qualidade dos frutos em anonáceas e produz frutos partenocárpicos em atemoieira 'Gefner'

ABSTRACT: The present research aimed to evaluate the effect of applying gibberellic acid (GA3) with hand (HP) or natural pollination (NP) on fruit set and the fruit quality of 'Red' and 'Lessard Thai' sugar apple and 'Gefner' atemoya fruits.This study was performed in an experimental orchard located in Homestead, Florida, USA. The experimental design included randomized blocks, with eight treatments, four replicates and four plants per plot. Treatments included: (1) HP; (2) HP + 10 mg L-1 GA3; (3) HP + 100 mg L-1 GA3; (4) HP + 1,000 mg L-1 GA3; (5) NP; (6) NP + 10 mg L-1 GA3; (7) NP + 100 mg L-1 GA3 and (8) NP + 1,000 mg L-1 GA3. The HP plus 1,000 mg L-1 GA3 promoted fruit setting above 90% over the 14 weeks for all genotypes evaluated. Significant increments for length and total fruit weight were observed. 'Red' sugar apple and atemoya had a reduced number of seeds per fruit. The NP plus GA3 (1,000 mg L-1) was effective in producing high quality seedless 'Gefner' atemoya fruits. This investigationdemonstratedthat GA3 plus hand pollination produced high quality sugar apple and atemoya seeded fruits and in association with natural pollination promoted seedless 'Gefner'atemoya fruits.

RESUMO: O presente trabalho teve como objetivo avaliar o efeito da aplicação de ácido giberélico (GA3) associado com e sem polinização artificial, no pegamento e na qualidade de frutos das pinheiras 'Red' e 'Lessard Thai' e da atemoieira'Gefner'. O experimento foi realizado em pomar localizado em Homestead, EUA. O delineamento experimental foi em blocos casualizados, com oito tratamentos, quatro repetições e quatro plantas por parcela. Os tratamentos utilizados neste experimento foram: 1) Polinização Artificial (PA); 2) PA + 10 mg L-1 GA3; 3) PA + 100 mg L-1 GA3; 4) PA + 1000 mg L-1 GA3; 5) Polinização natural (PN); 6) PN + 10 mg L-1 GA3; 7) PN + 100 mg L-1 GA3; e 8) PN + 1000 mg L-1 GA3. A PA combinada com a concentração de 1000 mg L-1 de GA3 promoveu o estabelecimento de frutos acima de 90% durante as 14 semanas de avaliação para todos os genótipos avaliados. Incrementos significativos para o comprimento e peso dos frutos também foram observados a variedade de pinha "Red" e a atemoia, em que apresentaram redução no número total de sementes.A PN em associação com GA3 (1000 mg L-1) foi eficaz na produção de frutos sem sementes de alta qualidade em atemóia 'Gefner. Estes dados mostram que o GA3, combinado com a polinização manual, produziu frutos com semente de alta qualidade e, em associação com a polinização natural, promoveu frutos sem sementes em atemóia 'Gefner'.

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.

Annonaceous acetogenins from seeds of Annona squamosa / 中草药

Objective To study the chemical composition from the seeds of Annona squamosa. Methods The components of annonaceous acetogenins from the seeds of A. squamosa were isolated and purified by extracting and chromatography separation. The structures of the compounds were identified by their physicochemical properties, UV, NMR, and mass spectrometry data. Results Seven annonaceous acetogenins were isolated from the ethyl acetate extract of the seeds from A.squamosa. They are 3-[7-[5- [1,4-dihydroxy-4-[5-(1-hydroxytridecyl)-2-furanyl] butyl] tetrahydro-2-furanyl]-2-hydroxyheptyl]-5-methyl-2(5H)-furanone (1), 3-[9-[5-(1-hydroxy-4-heptadeceneyl) tetrahydro-2-furanyl] hydroxynonyl]-5-methyl-2 (5H)-furanone (2), bullatalicin (3), squamostatin A (4), squamostatin D (5), bullatacin (6), and 10-hydroxyasimicin (7). Conclusion Compounds 1 and 2 are two new compounds, named as trilobalicin I (1) and D20-solamin B (2), respectively.

PCA and TOPSIS analysis and determination of amino acids and nucleosides in seeds of Annona squamosa from different producing areas by QTRAP-UFLC-MS/MS / 中草药

Objective To establish a QTRAP-UFLC-MS/MS method for the determination of 15 kinds of amino acids and 10 kinds of nucleosides in the seeds of Annona squamosa from different producing areas, toanalyze and investigate the difference of the 25 components in samples from five habitats, and provide scientific basis for its comprehensive utilization. Methods The analysis was carried out on a Waters XBridge Amide column (100 mm × 2.1 mm, 3.5 μm) with elution by mobile phase of 0.2% formic acid in water (A)-0.2% formic acid in acetonitrile (B) at a flow rate of 0.6 mL/min. The column temperature was maintained at 30 ℃. The target compounds were analyzed by the positive ion multiple reaction monitoring (MRM) mode. The comprehensive evaluation of the seeds from A. squamosa in five habitats was carried out by PCA and TOPSIS analysis. Results All 15 kinds of amino acids and 10 kinds of nucleosides showed good linearity (r2 > 0.994 7). The RSDs of the precision, repeatability, and stability tests were less than 3.87%. The average recovery rates were in the range from 94.90% to 98.78%, RSDs were in the range from 0.84% to 3.01%. There are differences in the composition and content of amino acids and nucleosides in the seeds of A. squamosa from different habitats, among which Hainan produced the highest comprehensive quality. Conclusion The method is simple, accurate, and reliable, which can provide the reliable basis for quality evaluation of the seeds from A. squamosa, and samples from different habitats can be optimized according to the target component.

Cytotoxicity and bacterial membrane destabilization induced by Annona squamosa L. extracts

ABSTRACT This study aimed to further investigate the cytotoxicity against tumor cell lines and several bacterial strains of Annona squamosa and its mode of action. Methanol extracts of A. squamosa leaves (ASL) and seeds (ASS) were used. ASL showed significant antibacterial activity against S. aureus, K. pneumoniae and E. faecalis with MIC values of 78, 78 and 39 µg/mL respectively. Moreover, ASL exhibited significant biofilm disruption, rapid time dependent kinetics of bacterial killing, increased membrane permeability and significantly reduced the cell numbers and viability. Regarding the cytotoxicity against tumor cell lines, ASS was more active against Jurkat and MCF-7 cells, with CI50 1.1 and 2.1 µg/mL, respectively. ASL showed promising activity against Jurkat and HL60, with CI50 4.2 and 6.4 µg/mL, respectively. Both extracts showed lower activity against VERO cells and reduced the clonogenic survival at higher concentrations (IC90) to MCF-7 and HCT-116 lineages. The alkaloids anonaine, asimilobine, corypalmine, liriodenine nornuciferine and reticuline were identified in extracts by UPLC-ESI-MS/MS analysis. This study reinforced that A. squamosa presents a remarkable phytomedicinal potential and revealed that its antimicrobial mechanism of action is related to bacterial membrane destabilization.

Two new annonaceous acetogenins from seeds of Annona squamosa / 中草药

Objective: To study the chemical constituents from the seeds of Annona squamosa. Methods: Modern methods such as silica gel column and preparation liquid phase were used to isolate and purify the components of annonaceous acetogenins from the seeds of A. squamosa. The structures of the compounds were identified by their physico-chemical properties, UV, NMR and mass spectrometry data. Results: Ten adjacent bistetrahydrofuran annonaceous acetogenins were isolated from CO2 supercritical extract of the seeds from A. squamosa. They were 3-((13R)-13-((2R,2'R,5R,5'R)-5'-((1S)-1,5-dihydroxynonyl) octahydro-[2,2'-bifuran]-5-yl)-13-hydroxytridecyl)-5-methylfuran-2 (5H)-one (1), 3-((2R,9R)-2,9-dihydroxy-9-((2R,2'R,5R,5'R)-5'-((S)-1-hydroxypentadecyl) octahydro-[2,2'-bifuran]-5-yl) nonyl)-5-methylfuran-2 (5H)-one (2), 6-hydroxy-desacetyl-uvaricin (3), 6-hydroxy-4-deoxy-squamotacin (4), bullanin (5), 10-hydroxy-asimicin (6), folianin A (7), annosquacin I (8), annosquacin C (9), and squamocin I (10). Conclusion: Compounds 1 and 2 are two new compounds named as 28-hydroxy-squamocin L (1) and 4-hydroxy-squamocin Y (2), respectively. Compounds 3-7 are isolated from A. squamosa for the first time.

Avaliação da atividade anti-Leishmania e anti-Trypanosoma do extrato etanólico das folhas de Annona squamosa L / Evaluation of the anti-leishmania and anti-trypanosoma activity of ethanolic extract from the leaves of Annona squamosa L

As doenças parasitárias, também chamadas de “doenças negligenciadas”, continuam sendo uma grande dificuldade para o desenvolvimento social e econômico dos países mais pobres. Podemos citar como exemplo dessas doenças, a leishmaniose e a doença de Chagas. A leishmaniose é causada por parasitas do gênero Leishmania e afeta cerca de 12 milhões de pessoas. A doença de Chagas, causada pelo protozoário Trypanosoma cruzi, causa aproximadamente 50.000 mortes por ano. Os fármacos disponíveis para o tratamento dessas doenças são altamente tóxicos, sendo este um dos motivos que leva à busca por drogas eficazes e seguras para seus tratamentos. As folhas da Annona squamosa, espécie da família Annonaceae, já foram descritas na literatura por suas atividades hepatoprotetora, antiparasitária, pesticida e antimicrobiana. Nesse estudo avaliamos a atividade anti-leishmania e tripanocida do extrato etanólico das folhas de Annona squamosa L. (EEAS) em formas promastigota do parasita Leishmania braziliensis e Leishmania infantum e epimastigota de Trypanosoma cruzi, além de avaliar a atividade citotóxica em fibroblasto. Os resultados demonstram que o extrato apresentou uma melhor atividade contra Leishmania infantum e Leishmania brasiliensis quando comparados com Trypanosoma cruzi; e que apresentou uma maior toxicidade nas concentrações de 500 e 1000 μg/ml, com mortalidade dos fibroblastos de aproximadamente 85% e 100%, respectivamente. Esse estudo aponta para uma perspectiva terapêutica alternativa que se mostrou eficaz frente aos parasitas aqui estudados, exceto a forma epimastigota de Trypanosoma cruzi. Com relação aos testes de citotoxicidade fazem-se necessários novos testes, uma vez que apresentou um alto nível de toxicidade, viabilizando assim futuros ensaios in vivo.

The parasitic diseases, also calls by “neglected diseases”, continue being a major difficulty for the social and economic development of the poorest countries. We can cite as an example of these diseases, the leishmaniasis and the Chagas disease. Leishmaniasis is caused by parasites of the genus Leishmania and affects about 12 million people. The Chagas disease, caused by the protozoan Trypanosoma cruzi, causes approximately 50,000 deaths per year. The drugs available for the treatment of these diseases are highly toxic, being this one of the reasons that leads to the search for effective and safe drugs for their treatments. The leaves of the Annona squamosa, species of the family Annonaceae, have already been described in the literature by their hepatoprotective activities, antiparasitic, pesticide and antimicrobial. In this study we assessed the activity tripanocidal and antileishmania of ethanolic extract from the leaves of Annona squamosa L. (EEAS) in promastigota forms of the parasite Leishmania braziliensis and Leishmania infantum and epimastigota of Trypanosoma cruzi, in addition to evaluating the cytotoxic activity in fibroblasts. The results demonstrate that the extract presented a better activity against Leishmania infantum and Leishmania brasiliensis when compared with Trypanosoma cruzi; and which presented a greater toxicity at concentrations of 500 and 1000 μg/ml, with mortality of fibroblasts of approximately 85% and 100%, respectively. This study points to an alternative therapeutic perspective that showed effective against the parasites here studied, except the epimastigota form of Trypanosoma cruzi. With relation to cytotoxicity tests are required new tests, once presented a high level of toxicity, thus enabling future in vivo assays.

Ocular toxicity by seeds of Annona squamosa (custard apple).

Custard apple seeds have been used in native medicine from time immemorial for the management of head lice and skin exfoliation. We report six consecutive patients who developed toxic keratoconjunctivitis within 6–12 h of ocular exposure to custard apple seeds. The use of topical steroid worsens the toxicity and predisposes to the development of microbial keratitis in such cases. Patients showed a good response to primary treatment with topical fortified antibiotics and lubricants. This case series highlights the need to educate the patients regarding the potential toxic effects of the custard seeds and the treating physicians about possible deleterious effects on using topical steroid.

Neuroprotective effect of Annona squamosa & (-) Anonaine in decreased GABA receptor of epileptic rats.

Annona squamosa and its active alkaloid content (-) Anonaine are known for antiepileptic activity on the basis of literature review. The objective of this study was to establish neuroprotection by Annona squamosa and its active alkaloid content (-) Anonaine as an antiepileptic agent by acting on GABA receptor. For neuroprotective assessment, variations of GABA, GABAA, GABAB receptors in the cortex region of the brain in epileptic rats and potential applications of Annona squamosa and its isolated chemical constituent (Anonaine) were investigated by using confocal microscopy. Nootropic activity in epileptic rats was also studied by radial and Y-maze model. GABA receptor binding analysis in the cortex region of epileptic rats showed significant decrease in Bmax (p<0.001) compared to control. Microscopic (confocal) study reveals confirmed decreased GABA receptors in epileptic rats. The extract of Annona squamosa leaves and its isolated constituent- anonaine showed memory boosting and memory regaining effects in radial arm and y-maze model. From the above data and findings, we may conclude that extract of Annona squamosa leaves and its alkaloidal constituent- anonaine improves significant changes in epileptic rat’s behavior and their decreased GABA receptors. Both extract and its isolated component in future may be evaluated in further studies.

Bioatividade dos extratos aquosos de plantas às larvas da mosca-das-frutas, Ceratitis capitata (Wied. ) / Bioactivity of aqueous extracts of plants to the fruit fly larvae, Ceratitis capitata (Wied. )

O objetivo deste trabalho foi avaliar a toxicidade dos extratos aquosos de plantas às larvas recém-eclodidas de Ceratitis capitata. Folhas de algodão-de-seda (Calotropis procera), citronela (Cymbopogon nardus), juazeiro (Ziziphus joazeiro), pinha (Annona squamosa),pinhão-manso (Jatropha curcas), noni (Morinda citrifolia) e tingui (Magoniapubescens) foram coletadas, secas e moídas em moinho de facas até a obtenção de um pó. Tais pós foram adiciona dos em água destilada para a preparação dos extratos aquosos (10%, p/v). As suspensões contendo os compostos hidrossolúveis foram filtradas, a fim de se conseguir os extratos que foram adicionados à dieta artificial das larvas da mosca. À dieta controle, adicionou-se somente água destilada. Os ensaios foram realizados em DIC, com quatro tratamentos e um controle. Todos os tratamentos foram constituídos por 10 repetições, cada uma com 10 larvas recém-eclodidas de C. capitata. Foram avaliadas a mortalidade larval e a eficiência de controle (E%) dos extratos aquosos às larvas. Os extratos de juazeiro, tingui e pinhão-manso são os mais tóxicos e eficientes no controle das larvas de C. capitata, causando elevadas mortalidades.(AU)

The objective of this study was to evaluate the toxicity of aqueous plant extracts to neonate larvae of Ceratitis capitata. Silk cotton (Calotropis procera), citronella grass (Cymbopogon nardus), joazeiro (Ziziphus joazeiro),sugar apple (Annona squamosa), jatropha (Jatropha curcas), noni (Morinda citrifolia) and tingui (Magonia pubescens) were collected, dried, and milled in a knife mill in order to obtain a powder. These powders were added in distilled water to prepare aqueous extracts (10%, w/v). The suspension containing the hydrosoluble compounds was filtered to get the extracts, which were added to the artificial diet of fruit fly larvae. Only distilled water was added in the control diet. Assays were carried out in DIC, with four treatments (aqueous extracts) and one control. All treatments consisted of 10 repetitions, each containing 10 newly hatched larvae ofC. capitata. Larval mortality and control efficiency (E%) of aqueous extracts to the larvae were evaluated. The joazeiro, tingui and jatropha extracts are the most toxic and effective in controlling larvae of C. capitata, causing high mortality.(AU)

Comparative Evaluation of Antioxidant Properties of Methanolic Extract of Red and Green Custard Apple Fruits.

Annona squamosa (custard apple) or also called Sugar Apple. It is native to the tropical Americas and West Indies, but the exact origin is unknown. It is now the most widely cultivated of all the species of Annona, being grown for its fruit throughout the tropics and warmer subtropics, such as Indonesia, Thailand, and Taiwan; it was introduced to southern Asia before 1590. There are two varieties of custard apple which were available in the local Malaysian market. In the present study we aimed to carry out comparative antioxidant activity of methanolic extract of both custard apple fruits by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) method. The results showed that the methanolic extract of red custard apple showed better antioxidant activity than methanolic extract of green custard apple. However, the standard ascorbic acid showed better antioxidant activity than both the extracts. In conclusion, the red custard apple is good candidate for further investigation.