Annona Crassiflora Research Articles

Stephalagine, an alkaloid with pancreatic lipase inhibitory activity isolated from the fruit peel of Annona crassiflora Mart.

Annona crassiflora Mart., a member of the Annonaceae family found in the Brazilian Savannah, is acknowledged as a plant rich in a variety of isoquinoline alkaloids. The isoquinoline skeleton-derivative alkaloids have been reported as pancreatic lipase (PL) inhibitors. However, the presence of these bioactive compounds in the fruit peel of A. crassiflora remains unclear. This study was conducted to find a bioactive alkaloid not yet described in the fruit peel of A. crassiflora and evaluate its capacity to inhibit the PL activity through in vitro kinetic assay using p-nitrophenyl palmitate as substrate. The ethanolic extract (EtOH) of the peels of A. crassiflora fruits was subjected to liquid−liquid extraction, which yielded an alkaloidal fraction. After a high performance liquid chromatography (HPLC) separation, the purified alkaloid was subjected to structural characterization by high-resolution mass spectrometry with electrospray ionization and electron impact, and nuclear magnetic resonance (NMR) spectroscopy using 1H, 13C, correlation spectroscopy (COSY), heteronuclear single-quantum correlation spectroscopy (HSQC), heteronuclear multiple-bond correlation spectroscopy (HMBC) and HMBC-N analyses. The metabolite stephalagine (1,2-methylenedioxy-3-methoxyaporphine), an aporphine alkaloid, was identified. The purified stephalagine was further evaluated as a PL inhibitor. Stephalagine showed high PL inhibitory activity (IC50 of 8.35μgmL−1) with low cytotoxicity, suggesting a potential anti-obesity agent. This is the first report of the isolation of stephalagine in Annonaceae and its inhibitory effect on pancreatic lipase activity.

Peel of araticum fruit (Annona crassiflora Mart.) as a source of antioxidant compounds with α-amylase, α-glucosidase and glycation inhibitory activities

Annona crassiflora Mart., whose fruit is popularly known as araticum, is a member of the Annonaceae family found in the Brazilian Cerrado. Although this plant has several medicinal uses, its bioactive molecules are not fully understood. A bioguided assay was performed to identify the main bioactive compounds of A. crassiflora fruit peel from the ethanol extract fractions with antioxidant capacity and α-amylase, α-glucosidase and glycation inhibitory activities. Ethyl acetate and n-butanol fractions showed, respectively, higher antioxidant capacity (DPPH IC50 1.5±0.1 and 0.8±0.1μgmL−1, ORAC 3355±164 and 2714±79μmoltroloxeq/g, and FRAP 888±16 and 921±9μmoltroloxeq/g) and inhibitory activities against α-amylase (IC50 4.5±0.8 and 1.7±0.3μgmL−1), α-glucosidase (IC50 554.5±158.6 and 787.8±140.6μgmL−1) and glycation (IC50 14.3±3.3 and 16.0±4.2μgmL−1), and lower cytotoxicity, compared to the other fractions and crude ethanol extract. The HPLC-ESI-MS/MS analysis identified various biomolecules known as potent antioxidants, such as chlorogenic acid, (epi)catechin, procyanidins, caffeoyl-hexosides, quercetin-glucosides and kaempferol. The fruit peel of A. crassiflora, a specie from Cerrado, the Brazilian Savanna, provided a source of antioxidant compounds with properties to block carbohydrate digestive enzymes and formation of glycation products. Thus, there is potential to use the by-products of araticum in order to identify and isolate phytochemicals for application in nutraceutical supplements, food additives and pharmaceuticals products.

196 - Profile of Antioxidant Activity of Fruits of the Savana Brazilian

Araca ( Psidium guineense Swartz .), Baru ( Dipteryx alata Vogel ), Guavira ( Campomanesia pubescens (DC.) O. Berg), Mangaba ( Hancornia speciosa Gomez ), Marolo ( Annona crassiflora Mart .) and Pequi ( Caryocar brasiliense Camb .), are typical fruits found in the biome of Savana Brazilian, known and consumed in different ways, mainly by the native population. This study investigated the profile of the antioxidant activity “in vitro” the pulp of the fruits above described. The radical scavenging activity (antioxidant activity) was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH), at different concentrations (µg/mL) from the lyophilized pulp extracts subjected to extraction with water for 24 h. Ascorbic acid was used as a standard positive control for comparison of results. The extracts of the investigated fruit pulps showed radical capture percentage (DPPH) represented by (IC 50 ) inhibitory concentration of 50% (Fig. 1). The extracts of the fruit pulps demonstrated extraordinary antioxidant profile in DPPH of manner concentration dependent, especially for fruits Araca (IC 50= 218.2±5.0 µg/mL) and Guavira (223,2±1.72 µg/mL). The growing profile is maintained as concentrations, this way the Pequi, the Marolo, the Mangaba and the Baru (IC 50= 979,9±2.29 µg/mL, IC 50= 1.955±1.32 µg/mL, IC 50= 2.021,7±2.13 µg/mL and IC 50= 3.592±0.93 µg/mL), respectively. The results show that the pulps of the fruits of the Brazilian Savana have antioxidant potential and possibly their bioactive properties can attenuate damage caused by oxidative stress. The use of fruits, obtained through diet or herbal medicine, accessible to the world's population, become a source of natural antioxidants. Figure 1: Profile of antioxidant activity of fruits of the Savana Brazilian. Support by: FUNDECT (Fundacao de Apoio ao Desenvolvimento do Ensino, Ciencia e Tecnologia do Estado de Mato Grosso do Sul) and CNPq (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico).

Peltatoside Isolated from Annona crassiflora Induces Peripheral Antinociception by Activation of the Cannabinoid System.

Peltatoside is a natural compound isolated from leaves of Annona crassiflora Mart., a plant widely used in folk medicine. This substance is an analogue of quercetin, a flavonoid extensively studied because of its diverse biological activities, including analgesic effects. Besides, a previous study suggested, by computer structure analyses, a possible quercetin-CB1 cannabinoid receptor interaction. Thus, the aim of this work was to assess the antinociceptive effect of peltatoside and analyze the cannabinoid system involvement in this action. The mouse paw pressure test was used and hyperalgesia was induced by intraplantar injection of carrageenan (200 µg/paw). All used drugs were administered by intraplantar administration in Swiss male mice (n = 6). Peltatoside (100 µg/paw) elicited a local inhibition of hyperalgesia. The peripheral antinociceptive action of peltatoside was antagonized by the CB1 cannabinoid antagonist AM251 (160 µg/paw), but not by CB2 cannabinoid antagonist AM630 (100 µg/paw). In order to assess the role of endocannabinoids in this peripheral antinociceptive effect, we used (i) [5Z,8Z,11Z,14Z]-5,8,11,14-eicosatetraenyl-methyl ester phosphonofluoridic acid, an inhibitor of anandamide amidase; (ii) JZL184, an inhibitor for monoacylglycerol lipase, the primary enzyme responsible for degrading the endocannabinoid 2-arachidonoylglycerol; and (iii) VDM11, an endocannabinoid reuptake inhibitor. MAFP, JZL184, and VDM11 did not induce antinociception, respectively, at the doses 0.5, 3.8, and 2.5 µg/paw, however, these three drugs were able to potentiate the peripheral antinociceptive effect of peltatoside at an intermediary dose (50 µg/paw). Our results suggest that this natural substance is capable of inducing analgesia through the activation of peripheral CB1 receptors, involving endocannabinoids in this process.

Genetic Diversity and Structure of Populations of Annona crassiflora Mart. of Brazilian Savanna and Its Association with Chemical Variability

Annona crassiflora Mart. is a native tree from Brazilian savanna. Isoquinoline alkaloids are characteristic of species of Annonaceae. This work aimed to assess the magnitude of genetic diversity among different populations of A. crassiflora using AFLP markers, and verify the existence of any correlation between the AFLP data and previous reported alkaloid composition. A. crassiflora from eight populations in the states of São Paulo, Goiás, Minas Gerais, and Distrito Federal were analyzed. The data suggest a low, moderate, and high level of genetic diversity from different populations of A. crassiflora. Concentration of alkaloids was significantly correlated with AFLP data, suggesting interaction between chemical and molecular markers in A. crassiflora. The data of association between the chemical and genetic differentiation of A. crassiflora may be useful to establish cultivation areas allowing the definition of strategies to preserve their genetic diversity with an interest in specific chemotypes for genetic improvement programs focused on sustainable utilization of this specie.

Climatic drivers of leaf traits and genetic divergence in the tree Annona crassiflora: a broad spatial survey in the Brazilian savannas.

The Cerrado is the largest South American savanna and encompasses substantial species diversity and environmental variation. Nevertheless, little is known regarding the influence of the environment on population divergence of Cerrado species. Here, we searched for climatic drivers of genetic (nuclear microsatellites) and leaf trait divergence in Annona crassiflora, a widespread tree in the Cerrado. The sampling encompassed all phytogeographic provinces of the continuous area of the Cerrado and included 397 individuals belonging to 21 populations. Populations showed substantial genetic and leaf trait divergence across the species' range. Our data revealed three spatially defined genetic groups (eastern, western and southern) and two morphologically distinct groups (eastern and western only). The east-west split in both the morphological and genetic data closely mirrors previously described phylogeographic patterns of Cerrado species. Generalized linear mixed effects models and multiple regression analyses revealed several climatic factors associated with both genetic and leaf trait divergence among populations of A.crassiflora. Isolation by environment (IBE) was mainly due to temperature seasonality and precipitation of the warmest quarter. Populations that experienced lower precipitation summers and hotter winters had heavier leaves and lower specific leaf area. The southwestern area of the Cerrado had the highest genetic diversity of A.crassiflora, suggesting that this region may have been climatically stable. Overall, we demonstrate that a combination of current climate and past climatic changes have shaped the population divergence and spatial structure of A.crassiflora. However, the genetic structure of A.crassiflora reflects the biogeographic history of the species more strongly than leaf traits, which are more related to current climate.

Characterization of chemical and mineral composition of marolo (Annona crassiflora Mart) during physiological development

Maroleiro (Annona crassiflora Mart), belonging to the family of Annonaceae is a typical specie of the Cerrado and has occurred in most states. For fruit characteristics “sui generis”, with significant nutritional value. This study aimed to characterize the marolo (Anonna crasiflora Mart) during fruit development, through the evaluation of its proximal and mineral composition. The fruits 2012/2013 season, were picked to 12Km from the city of Itumirim, southern Minas Gerais, in intervals of 20 days from anthesis to maturity, and the first fruits were harvested 60 days after anthesis. The flowering marolo began in late September 2012, with fruiting initially observed at the end of October, with peak production at the end of February and beginning of March 2013 The fruits were harvested in the morning and carried to the laboratory, where followed up the moisture, ether extract, protein, ash, fiber and high sugar fraction beyond mineral quantification of them. During development reductions in moisture content and protein increments of ether extract, crude fiber and high sugar fraction and an oscillation in the mineral along the developmental stages was observed.

Species-specific phylogeographical patterns and Pleistocene east-west divergence inAnnona(Annonaceae) in the Brazilian Cerrado

The Brazilian Cerrado is the most diverse savanna in the world. Phylogeographical patterns for Cerrado tree species are relatively congruent; however, the causes of these patterns are not clear. Ours aims were to investigate phylogeographical patterns in two co-distributed Annona spp. in the Cerrado and identify the factors responsible for their genetic structure. We sequenced non-coding plastid DNA regions and estimated divergence times using a Bayesian Markov chain Monte Carlo method implemented in BEAST. Generalized linear mixed models (GLMMs) were used to relate genetic divergence with spatial, climatic and phytogeographic data. Annona crassiflora possessed only one haplotype, whereas A. coriacea possessed nine haplotypes constituting two groups: western and eastern. Haplotype diversification first occurred in the Pliocene; the split between eastern and western groups was dated to the Early Pleistocene. GLMMs detected a weak association of genetic distance with current climatic factors, represented mainly by precipitation in the warmest months. In A. coriacea, the north of central Cerrado (around Serra Geral de Goias) is suggested as a putative Pleistocene refugium. The current climate and dispersal and local adaptation processes probably contribute to maintaining the east–west genetic structure in A. coriacea. Both Annona spp. show idiosyncratic population histories, reminiscent of the complex biogeography of the Cerrado.

Effect of adding flours from marolo fruit (Annona crassiflora Mart) and jerivá fruit (Syagrus romanzoffiana Cham Glassm) on the physicals and sensory characteristics of food bars

The marolo (Annona crassiflora Mart.) and jeriva (Syagrus romanzoffiana Cham Glassm) fruits grow in the Cerrado biome, and have important sensory and nutritional characteristics. These fruits are eaten fresh or processed and embedded in ice cream, candy, juices, and liquors. However, their use is very limited and is localized exclusively to their native region. The aim of this study was to evaluate the use of these fruits in food consumer products to improve the nutritional patterns of consumers and extend knowledge thereof. To do this, different levels (0%, 5%, 10%, 15%, and 20%) of flour derived from the marolo and jeriva fruits were incorporated into food bars, which were then characterized in terms of their physical of the texture (TPA), porosity, color and sensorial properties. The food bars enriched with marolo and jeriva flours had an intense yellow color and increased hardness relative to the control bars; the porosity of the bars progressively decreased as the amount of flour added increased. The sensorial analysis in the snack bars developed added with the fruits flour no was influence by incorporation this component, showed good averages for the scores evaluated.

English

Researches have shown that marolo (Annona crassiflora Mart.) contains many biologically active phytochemical classes with antidiabetic properties. Therefore, in order to obtain scientific data that contribute to a better insight of the species, integrating both safety and efficacy to the practice of using the plant as medicine, the study of A. crassiflora Mart. leaves was chosen. To evaluate the possible effect of A. crassiflora Mart. in diabetic rats, blood glucose levels, concentrations of fructosamine, urea, creatinine, ASL and ALT was determined in serum samples from animals treated with 300 mg of hydroethanolic extract 70% (v/v) per kg of animal weight. Glucose levels were determined by Trinder reaction. Dosages of urea and creatinine were held using the Jaffe method for creatinine and the enzyme method colorimetric for urea in automated apparatus (HumaStar 80, Brazil/Alfenas). The concentration of fructosamine was measured by a commercial kit kinetic method (analyzes ®) and the reading was held in Bioplus® (Brazil- Alfenas). Based on the results obtained in this study, it can be concluded that administrating the extract, according to the adopted protocol, did not change the biochemical parameters that were analyzed in the normal and diabetic rats’ serums. Key words: Medicinal plants, phytochemical, diabetes mellitus, pharmacological, hydroethanolic extract.

Population structure of Annona crassiflora: an endemic plant species of the Brazilian Cerrado.

Habitat fragmentation has numerous consequences, particularly to endemic species, and has a negative impact on the genetic diversity of neglected species, leading to genetic drift. Annona crassiflora Mart. is a species that is endemic to Brazil, and its incidence in the Cerrado biome has decreased. The identification and characterization of its remaining diversity is necessary for its conservation. Our aim was to study the population structure of A. crassiflora populations from different Cerrado regions in Minas Gerais State, Brazil (Corinto, Curvelo, Carmo da Mata, Boa Esperança, and Paraguaçu) using inter-simple sequence repeat (ISSR) markers and DNA content. Nuclear DNA content was estimated by flow cytometry using 10 individuals from each population. ISSR markers were used for genotyping accessions in order to study their genetic diversity and population structures. We found considerable genetic variation among populations, with the highest variability observed in the Curvelo population. There was a significant positive correlation between DNA content and latitude (r = 0.46, P = 0. 0003). A Bayesian-based cluster analysis grouped the populations into three clusters, which followed their geographical origins. There was some level of genetic diversity and differentiation among the populations, suggesting the need for a conservation plan for this species. The ISSR markers and DNA content analysis were effective in studying the genetic diversity and population structure of A. crassiflora.

Development and sensory evaluation of products containing the Brazilian Savannah fruits araticum (Annona crassiflora Mart.) and cagaita (Eugenia dysenterica Mart.)

Summary The objective of this study was to use the pulp of the Brazilian Savannah fruits araticum and cagaita to develop new products, and to evaluate their sensory acceptance by children from 7 to 9 years of age. The products developed were: three milk caramel formulations: DP (standard), DA (araticum pulp) and DB (araticum pulp and grated coconut); three cagaita refresher formulations: RP (cagaita pulp), RA (cagaita pulp and orange juice) and RB (cagaita pulp and carrot juice); and a cagaita jam formulation. It was found that the three formulations of milk caramels were equally accepted (p>0.05), whereas the RA refresher was more accepted (p<0.05) than the others (RP and RB). In addition, the cagaita jam received high sensory acceptance scores. All the products developed were well accepted since their acceptability indexes were greater than 70%. There was no influence (p>0.05) of gender on the sensory acceptability of the products evaluated. It was concluded that the products developed showed good acceptability from a sensory point of view, which suggests high market potential.

Fenologia reprodutiva do araticum e suas implicações no potencial produtivo

The araticum (Annona crassiflora Mart.) is a native fruit from the Cerrado biome, with a great economic and social potential. Studies on Cerrado native plants phenology are scarce and can be promising for a better rational and sustainable exploitation of its potentialities. Thus, the objective of this study was to characterize araticum reproductive phenology in the Cerrado. The phenophases evaluations were carried out from August 2008 to August 2010, in Goiania, Goias, Brazil, through counting of flower buds, flowers in anthesis and fruits formed. The production capacity was estimated through the effective fruiting rate. For flower emission, a biannual pattern was observed for araticum.. The effective fruiting rate observed was 3.29% and the araticum’s reproductive phenophases were concentrated during the rainy season. The effective fruiting rate is considered very low and depends on biotic and abiotic factors.

Português

The biopanning of new molecules is of great importance due to the limitation on the efficacy or toxicit of the main therapeutic agents available for the treatment of diseases. In recent years, the indiscriminate use of antibiotics has led microorganisms develop resistance, making it necessary new technologies. Thus, the native medicinal plants have been a key means for the pursuit of bioactive substances. This study aimed to evaluate the antimicrobial activity of extracts Aspidosperma nitidum, Annona crassiflora and Annona mucosa, against strains Staphylococcus aureus ATCC® 25923, Escherichia coli ATCC® 25922 and Pseudomonas aeruginosa ATCC® 27853. The extract was diluted in each plant concentrations of 1:2, 1:4, 1:8, 1:16, 1:32. Of each bacterial strain were selected 3 to 4 colonies, which were transferred to test tubes containing NaCl solution, and subjected to serial dilution to obtain a microorganism load UFC/ml 10-8, and incubated at 35°C for 5 hours. Each of the three mi cro-organism has been subjected to the three extracts and extract five treatment with 5 replicates for each concentration of the extracts. With the aid of a caliper measurements were performed halos of inhibition of bacterial growth. Whereas inhibition of microbial activity the solutions presented diameters greater than 8 mm. The extract of the three plants presented satisfactory results in the control of the three agencies. The extract of Annona mucosa presented in all organisms analyzed average inhibitory values greater than the other extracts. All statements used in this study semonstraram great potential antimicrobials.

Biological activities of Annonaceae species extracts from Cerrado

Annonaceae is one of the richest families of the wood components of the Brazilian savanna (Cerrado). Annona coriacea Mart., Annona crassiflora Mart., Duguetia furfuracea (A. St.-Hil.) Saff. and Xylopia aromatica L. are common species in this region. The main goal of our study was to determine biological activities of these species. Therefore, we evaluated phytotoxic effects of ethanolic extracts of leaves and stems of these species in the laboratory. In addition, toxicity to animal targets was also evaluated. Lettuce was the less affected target species, with discreet effects over germination rate and initial root development observed using the highest concentration (0.8 mg mL−1) of extracts of A. coriacea. Notwithstanding, tomato was the most affected target species by these annonaceous extracts, with an expressive decrease in germination rate, shoot and root growth even with the lowest extract concentrations. Onion germination was affected mainly by X. aromatica and A. coriacea leaf extracts, with significant results for all concentrations. Effects on shoot and root growth were also observed for this species. In general, the strongest phytotoxic activities were detected for the extracts of A. coriacea and X. aromatica. These extracts also inhibited growth of Urochloadecumbens (Stapf) R.D. Webster. The extracts of A. coriacea were also very toxic to brine shrimp nauplii. No antibacterial or antifungal effects were detected with the extracts of these four species of Annonaceae. Among the four species, X. aromatica and A. coriacea exhibited strong biological activities and the results could be useful in agricultural fields in Brazil. These extracts were also promising for isolation and identification of commercial valuable metabolites.

Bioactive Compounds Found in Brazilian Cerrado Fruits.

Functional foods include any natural product that presents health-promoting effects, thereby reducing the risk of chronic diseases. Cerrado fruits are considered a source of bioactive substances, mainly phenolic compounds, making them important functional foods. Despite this, the losses of natural vegetation in the Cerrado are progressive. Hence, the knowledge propagation about the importance of the species found in Cerrado could contribute to the preservation of this biome. This review provides information about Cerrado fruits and highlights the structures and pharmacologic potential of functional compounds found in these fruits. Compounds detected in Caryocar brasiliense Camb. (pequi), Dipteryx alata Vog. (baru), Eugenia dysenterica DC. (cagaita), Eugenia uniflora L. (pitanga), Genipa americana L. (jenipapo), Hancornia speciosa Gomes (mangaba), Mauritia flexuosa L.f. (buriti), Myrciaria cauliflora (DC) Berg (jabuticaba), Psidium guajava L. (goiaba), Psidium spp. (araçá), Solanum lycocarpum St. Hill (lobeira), Spondias mombin L. (cajá), Annona crassiflora Mart. (araticum), among others are reported here.

English

The Brazilian cerrado has several fruit species with great potential for agro-industrial use including the production of milk drinks. Thus, the objective of this study was to evaluate the sensory profile and purchase intent of fermented milk drinks added in increasing levels of fruits from the Brazilian cerrado. Four formulations of milk drinks were processed with concentrations of 4, 8, 12 and 16% of Araca (Psidium cattleianum), Araticum (Annona crassiflora Mart), Mangaba (Hancornia speciosa Gomes), Passion fruit (Passiflora edulis f. flavicarpa) and Pequi pulps (Caryocar brasiliense Camb.). The sensory profile of products was characterized by affective test evaluating acceptance using the hedonic scale and purchase intent. Fruits used in flavoring, especially Pequi, are very appreciated by the local population. As demonstrated in acceptability tests, it is believed that this factor contributed to the high acceptance of Pequi. All milk drinks showed positive purchase intent value. Key words: Fermented milk, acceptance test, purchase intent.

Analysis of the anti-inflammatory and chemopreventive potential and description of the antimutagenic mode of action of the Annona crassiflora methanolic extract

Context: Annona crassiflora Mart. (Annonaceae) is a medicinal plant that is widely used in folk medicine, which leads to its investigation as a potential source of new pharmacological principles.Objective: This study describes the anti-inflammatory, antiallodynic, and antimutagenic/chemopreventive activities of the leaves A. crassiflora methanolic extract. Its antimutagenic mode of action was analyzed in a plant or animal experimental model.Materials and methods: Total flavonoids were quantified by spectrophotometry at 415 nm and its composition was analyzed by 1H NMR spectra. Animals received orally, 30, 100, and 300 mg/kg of extract in both tests, carrageenan-induced paw edema and myeloperoxidase activity. Animals were treated with 100 and 300 mg/kg, in all the analyzed tests, pleural cell migration and protein exudation, carrageenan-induced cell migration into the pouch, induction of joint inflammation and carrageenan-induced allodynia response in the mouse paw. To evaluate the antimutagenic/chemopreventive activity through the Allium cepa test, we used 5, 10, and 15 mg/L of extract, and for the micronucleus test in the peripheral blood, we used the dose of 15 mg/kg.Results: The fractionation of the ethyl acetate (EA) fraction, resulting from the partition of the methanol extract of the A. crassiflora, afforded through chromatographic methods resulted in the isolation of kaempferol 3-O-β-glucoside and kaempferol 3-O-β-diglucoside. Oral treatment with 100 and 300 mg/kg of extract significantly inhibited the carrageenan-induced edema formation, with inhibitions of 53 ± 7% and 47 ± 10%; in MPO activity, the observed inhibitions were 60 ± 7% for 100 mg/kg treatment and 63 ± 7% for 300 mg/kg. The ACME reduced significantly the total leukocytes (an inhibition of 78 ± 9% with 100 mg/kg and 90 ± 7% with 300 mg/kg) and protein levels (approximately 100% inhibition with both doses) in the pleurisy model. In carrageenan-induced leukocyte migration into the pouch, the extract inhibited leukocyte migration only when administered 300 mg/kg per dose (the reduction was 43 ± 5%). Pretreatment with extract failed to reduce the zymosan-induced edema formation and did not inhibit the carrageenan-induced mechanical allodynia. Damage reduction in Allium cepa tested with different concentrations (5, 10, and 15 mg/L) was 66.17, 75.75, and 69.19% for the pre-treatment; 72.72, 33.33, and 22.22% for the simple simultaneous treatment; 100.50, 93.93, and 102.52% for the simultaneous treatment with pre-incubation; 89.39, 79.79, and 84.34%; for the post-treatment, and 86.36, 81.31, and 93.43% for the continuous treatment. The antimutagenic evaluation in the micronucleous test showed a damage reduction of 75.00 and 64.58% for the pre-treatment and simultaneous protocols, respectively. The post-treatment protocol increased the cyclophosphamide effects in 45.83%.Conclusion: These results suggest that this medicinal plant has chemopreventive and anti-inflammatory therapeutic potential.

In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae.

The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.

English

One hundred and twelve (112) endophytic fungi was isolated from leaf fragments of Annona crassiflora. Extracts were prepared by fermentation in Czapek broth. The extract of endophytic fungus Rhizoctonia sp. was selected as the most promising from an antimicrobial screening and its ability to produce metabolites with antimicrobial and antiproliferative activity was evaluated. Antimicrobial activity was evaluated through agar diffusion and broth microdilution. In the screening, the extract was able to inhibit Staphylococcus aureus, Escherichia coli and Candida albicans with inhibition zone diameters ranging from 27 to 40, but its antimicrobial activity was most prominent against S. aureus, with a minimum inhibitory concentration (MIC) of 500 to 1000 μg/ml. Regarding anti-proliferative activity, the results were promising. The extract displayed potent anti-proliferative activity against all tumor cell lines tested, suggesting the potential capacity to diminish the growth of human tumor cells, while also demonstrating selectivity by displaying reduced activity against non-tumor cell lines. Key words: Endophytic fungi, Annona crassiflora, fermentation process, bioactive compounds, biological assays.