Ontogenesis of Resin Ducts and Secretory Process in Protium spruceanum (Burseraceae) Stems.

The objective of this work was to characterize the ontogenesis of Protium spruceanum secretory ducts, to evaluate the effects of seasonality on that process, and to characterize the chemical nature of the resin. Morphometric, anatomical, micromorphometric, histochemical, and ultrastructural evaluations of shoot apexes and chemical analyses of the resin were performed. The ducts of schizolysigenous origin are distributed in the primary and secondary phloem. The subsecretory tissue is meristematic and can restore the secretory epithelium. Secretory epithelial cells have wall thickening resembling that of the Casparian strip that regulates secretion reflux. The main resin compounds are pentacyclic triterpenoids, α- and ß-amyrins, and α- and ß-amyrenones, which are reported here for the first time for this species. The presence of electron-dense and electron-opaque structures, in the secretory epithelial cells, are compatible with the triterpenes and mucilage identified in the resin. Rising temperatures, rainfall, and increasing day length induce the formation of ducts in the vascular cambium throughout Spring/Summer. The abundant production of resin rich in pentacyclic triterpenes indicates the potential use of the species for medicinal and cosmetic purposes. The understanding that secretory processes are concentrated during the Spring/Summer seasons will contribute to the definition of resin extraction management strategies.

Phytochemicals of Avocado Residues as Potential Acetylcholinesterase Inhibitors, Antioxidants, and Neuroprotective Agents.

Avocado (Persea americana) is a widely consumed fruit and a rich source of nutrients and phytochemicals. Its industrial processing generates peels and seeds which represent 30% of the fruit. Environmental issues related to these wastes are rapidly increasing and likely to double, according to expected avocado production. Therefore, this work aimed to evaluate the potential of hexane and ethanolic peel (PEL-H, PEL-ET) and seed (SED-H, SED-ET) extracts from avocado as sources of neuroprotective compounds. Minerals, total phenol (TPC), total flavonoid (TF), and lipid contents were determined by absorption spectroscopy and gas chromatography. In addition, phytochemicals were putatively identified by paper spray mass spectrometry (PSMS). The extracts were good sources of Ca, Mg, Fe, Zn, ω-6 linoleic acid, and flavonoids. Moreover, fifty-five metabolites were detected in the extracts, consisting mainly of phenolic acids, flavonoids, and alkaloids. The in vitro antioxidant capacity (FRAP and DPPH), acetylcholinesterase inhibition, and in vivo neuroprotective capacity were evaluated. PEL-ET was the best acetylcholinesterase inhibitor, with no significant difference (p > 0.05) compared to the control eserine, and it showed neither preventive nor regenerative effect in the neuroprotection assay. SED-ET demonstrated a significant protective effect compared to the control, suggesting neuroprotection against rotenone-induced neurological damage.

Antiviral fungal metabolites and some insights into their contribution to the current COVID-19 pandemic.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak, which started in late 2019, drove the scientific community to conduct innovative research to contain the spread of the pandemic and to care for those already affected. Since then, the search for new drugs that are effective against the virus has been strengthened. Featuring a relatively low cost of production under well-defined methods of cultivation, fungi have been providing a diversity of antiviral metabolites with unprecedented chemical structures. In this review, we present viral RNA infections highlighting SARS-CoV-2 morphogenesis and the infectious cycle, the targets of known antiviral drugs, and current developments in this area such as drug repurposing. We also explored the metabolic adaptability of fungi during fermentation to produce metabolites active against RNA viruses, along with their chemical structures, and mechanisms of action. Finally, the state of the art of research on SARS-CoV-2 inhibitors of fungal origin is reported, highlighting the metabolites selected by docking studies.

Matteucinol, isolated from Miconia chamissois, induces apoptosis in human glioblastoma lines via the intrinsic pathway and inhibits angiogenesis and tumor growth in vivo.

Gliomas account for nearly 70% of the central nervous system tumors and present a median survival of approximately 12-17 months. Studies have shown that administration of novel natural antineoplastic agents is been highly effective for treating gliomas. This study was conducted to investigate the antitumor potential (in vitro and in vivo) of Miconia chamissois Naudin for treating glioblastomas. We investigated the cytotoxicity of the chloroform partition and its sub-fraction in glioblastoma cell lines (GAMG and U251MG) and one normal cell line of astrocytes. The fraction showed cytotoxicity and was selective for tumor cells. Characterization of this fraction revealed a single compound, Matteucinol, which was first identified in the species M. chamissois. Matteucinol promoted cell death via intrinsic apoptosis in the adult glioblastoma lines. In addition, Matteucinol significantly reduced the migration, invasion, and clonogenicity of the tumor cells. Notably, it also reduced tumor growth and angiogenesis in vivo. Moreover, this agent showed synergistic effects with temozolomide, a chemotherapeutic agent commonly used in clinical practice. Our study demonstrates that Matteucinol from M chamissois is a promising compound for the treatment of glioblastomas and may be used along with the existing chemotherapeutic agents for more effective treatment.

Anti-inflammatory and antinociceptive properties of the hydroalcoholic fractions from the leaves of Annona crassiflora Mart. in mice.

BACKGROUND: Annona crassiflora Mart., popularly known as "Araticum", is a native tree of the Brazilian Cerrado used in folk medicine for treatment of pain and inflammatory diseases. We proposed to analyze analgesic and anti-inflammatory properties of the filtrate (F1) and the precipitate (F2) of the hydroalcoholic fraction from the leaves of Annona crassiflora Mart. in mice. MATERIALS AND METHODS: Swiss mice were submitted to formalin-induced nociception test and tail-flick reflex test, to assess antinociceptive properties, and to the rota-rod test, for motor performance analyses. To evaluate anti-inflammatory properties, F1 and F2 were orally administered 1 h prior to the intrathoracic injection of carrageenan, zymosan, LPS, CXCL8, or vehicle in Balb/c mice and neutrophil infiltration was evaluated 4 h after injection. RESULTS: F1 and F2 reduced the licking time in the second phase of formalin-induced nociception test, but only F2 showed a dose-dependent response. Neither F1 nor F2 reduced the latency time in the tail-flick reflex test. In addition, motor performance alteration was not observed in F1- or F2-treated mice. F2 treatment significantly inhibited the neutrophilia induced by carrageenan, LPS, or CXCL8, but not zymosan. CONCLUSIONS: The experimental data demonstrated that hydroalcoholic fractions of Annona crassiflora Mart. leaves have remarkable anti-inflammatory and antinociceptive activities.

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.

Cuticular wax composition contributes to different strategies of foliar water uptake in six plant species from foggy rupestrian grassland in tropical mountains.

The cuticle is the outermost region of the epidermal cell wall of plant aerial organs. The cuticle acts as a two-way lipid barrier for water diffusion; therefore, it plays a vital role in foliar water uptake (FWU). We hypothesised that the chemical composition of the cuticular waxes influences the FWU strategy that plants adopt in a foggy tropical ecosystem. We analysed the leaf cuticular waxes of six plant species known by their different FWU strategies, in both qualitative and quantitative approaches, to test this hypothesis. We also investigated the fine structure of the plant cuticle by scanning electron microscopy. Neither the total wax loads nor the amounts of single wax compound classes correlated to the FWU. In contrast, the qualitative chemical composition of the cuticular waxes was related to the water absorption speed but not to the maximum water absorbed. The presence of wax crystals might interfere with the FWU. Our findings suggest that a complex three-dimensional network of the cuticular compounds contributes to different strategies of FWU in six plant species from foggy tropical mountaintops.

Metabolic Profile Changes in Mangalarga Marchador Horses Subjected to A Hypercaloric Diet Evaluated by Proton NMR Spectroscopy.

The incidence of equine metabolic syndrome, a condition that results in endocrinopathic laminitis, is increasing worldwide. Although it is well known that the development of this syndrome depends on imbalances in energy metabolism and genetic traits, its pathophysiology remains unclear. Hydrogen nuclear magnetic resonance (1H NMR) is a powerful tool used to compare metabolic profiles and to discriminate metabolites in living beings. The aim was to apply this technology to detect blood biomarkers candidates in horses that were previously demonstrated to developed metabolic changes when subjected to induced obesity. Nine Mangalarga Marchador horses received a hypercaloric diet for 5 months and serum metabolomic analysis was performed before, during, and after the diet period. The 1H NMR results were subjected to multivariate analysis and NMR analysis allowed to identify six compounds (alanine, threonine, choline, α-glucose, ß-glucose, and creatinine), and observe the increasing choline level over the assessment period in four animals. A hypercaloric diet altered the metabolic profile of horses, with an individual bias in the time at which these changes occurred. This study is the first to describe metabolomic compounds in Mangalarga Marchador horses subjected to a hypercaloric diet rich in non-structural carbohydrates.

Cellulose acetate nanofibers loaded with crude annatto extract: Preparation, characterization, and in vivo evaluation for potential wound healing applications.

In this study we prepared annatto-loaded cellulose acetate nanofiber scaffolds and evaluated both in vitro cytotoxicity and potential for wound healing in a rat model. Annatto extract, which has been used to accelerate wound healing, was added to cellulose acetate polymer and the resulting material was used to produce nanofiber scaffolds via electrospinning. Physicochemical, and thermal evaluation of the resulting nanofiber mats showed that incorporating annatto did not significantly affect the thermal or chemical stability of the polymer. Annatto extract did not demonstrate cytotoxicity in the HET-CAM assay or MTT assay for fibroblast culture. Scanning electron microscopy of the fibroblasts confirmed that cells spread and penetrated into the nanofiber. In vivo experiments confirmed that cellulose acetate retained its biocompatibility when associated with crude annatto extract, and suggested that dose/response modulation occurs between the annatto-functionalized nanofibers and mast cells, indicating the potential of this material for wound healing applications.

Ketamine can be produced by Pochonia chlamydosporia: an old molecule and a new anthelmintic?

BACKGROUND: Infection by nematodes is a problem for human health, livestock, and agriculture, as it causes deficits in host health, increases production costs, and incurs a reduced food supply. The control of these parasites is usually done using anthelmintics, which, in most cases, have not been fully effective. Therefore, the search for new molecules with anthelmintic potential is necessary. METHODS: In the present study, we isolated and characterized molecules from the nematophagous fungus Pochonia chlamydosporia and tested these compounds on three nematodes: Caenorhabditis elegans; Ancylostoma ceylanicum; and Ascaris suum. RESULTS: The ethyl acetate extract showed nematicidal activity on the nematode model C. elegans. We identified the major substance present in two sub-fractions of this extract as ketamine. Then, we tested this compound on C. elegans and the parasites A. ceylanicum and A. suum using hamsters and mice as hosts, respectively. We did not find a difference between the animal groups when considering the number of worms recovered from the intestines of animals treated with ketamine (6 mg) and albendazole (P > 0.05). The parasite burden of larvae recovered from the lungs of mice treated with ketamine was similar to those treated with ivermectin. CONCLUSIONS: The results presented here demonstrate the nematicidal activity of ketamine in vitro and in vivo, thus confirming the nematicidal potential of the molecule present in the fungus P. chlamydosporia may consist of a new method of controlling parasites.

Nematicidal activity of Annona crassiflora leaf extract on Caenorhabditis elegans.

BACKGROUND: The aim of this work was to investigate the potential nematicidal activity of Annona crassiflora leaf extract against Caenorhabditis elegans. METHODS: The hydroalcoholic leaf extract and its fractions (dichloromethane, ethyl acetate, methanol and water) were submitted to mobility assay against the roundworm Caenorhabditis elegans. GC-MS and NMR analysis were performed in order to identify metabolites. RESULTS: The dichloromethane and ethyl acetate fractions showed to be the most active among the hydroalcoholic leaf extracts and its four fractions. The percentages of C. elegans larvae immobility were 98.13 and 89.66%, respectively, at a concentration of 1000 µg.mL(-1). Besides some amino acids, palmitic acid methyl ester, 2-isopropyl-5-methylcyclohexanol, oleic acid methyl esther, stearic acid methyl ester, quercetin and kaempferol were also identified in these fractions. CONCLUSION: The results indicated that of A. crassiflora leaf ethanolic extract has a good potential as a source for natural nematicide.

The first report on flavonoid isolation from Annona crassiflora Mart.

Annona crassiflora, a native tree from Brazilian Cerrado, is reported to possess several ethnomedical uses. Here, we report the isolation and unambiguous characterisation of the flavonoids quercetin-3-O-ß-D-glucopyranosil(1 â†’ 6)-O-α-L-arabinoside (1), known as peltatoside, kaempferol-3-O-ß-D-galactopyranoside (2), quercetin-3-O-ß-D-galactopyranoside (3), quercetin-3-O-ß-L-arabinopiranoside (4) and the ( - )-epicatechin (5) from the hydroalcoholic portion of the leaf ethanolic extract. Their structures were elucidated by using 1D and 2D NMR, ESI-MS, UV/Vis spectroscopy, optical rotation analysis and literature data comparison. The leaf ethanolic extract and its isolated compounds were evaluated by using antimicrobial, antioxidant and larvicidal assays, expressing antimicrobial and antioxidant activities. This is the first report on flavonoid isolation from A. crassiflora.

In vivo antimalarial efficacy of acetogenins, alkaloids and flavonoids enriched fractions from Annona crassiflora Mart.

Annona crassiflora and Annonaceae plants are known to be used to treat malaria by traditional healers. In this work, the antimalarial efficacy of different fractions of A. crassiflora, particularly acetogenin, alkaloids and flavonoid-rich fractions, was determined in vivo using Plasmodium berghei-infected mice model and toxicity was accessed by brine shrimp assay. The A. crassiflora fractions were administered at doses of 12.5 mg/kg/day in a 4-day test protocol. The results showed that some fractions from woods were rich in acetogenins, alkaloids and terpenes, and other fractions from leaves were rich in alkaloids and flavonoids. The parasitaemia was significantly (p < 0.05, p < 0.001) reduced (57-75%) with flavonoid and alkaloid-rich leaf fractions, which also increased mean survival time of mice after treatment. Our results confirm the usage of this plant in folk medicine as an antimalarial remedy.

In vitro antifungal activity of fatty acid methyl esters of the seeds of Annona cornifolia A.St.-Hil. (Annonaceae) against pathogenic fungus Paracoccidioides brasiliensis.

INTRODUCTION: Fatty acids are abundant in vegetable oils. They are known to have antibacterial and antifungal properties. METHODS: Antifungal susceptibility was evaluated by broth microdilution assay following CLSI (formerly the NCCLS) guidelines against 16 fungal strains of clinical interest. RESULTS: In this work, fatty acid methyl esters (FAME) was able to inhibit 12 clinical strains of the pathogenic fungus Paracoccidioides brasiliensis and were also active in the bioautographic assay against Cladosporium sphaerospermum. CONCLUSIONS: FAME was a more potent antifungal than trimethoprim-sulfamethoxazole against P. brasiliensis under the experimental conditions tested.

In vitro antifungal activity of fatty acid methyl esters of the seeds of Annona cornifolia A.St.-Hil. (Annonaceae) against pathogenic fungus Paracoccidioides brasiliensis / Atividade antifúngica in vitro dos ésteres metílicos de ácidos graxos das sementes de Annona cornifolia A.St.-Hil. (Annonaceae) sobre o fungo patogênico Paracoccidioides brasiliensis

INTRODUCTION: Fatty acids are abundant in vegetable oils. They are known to have antibacterial and antifungal properties. METHODS: Antifungal susceptibility was evaluated by broth microdilution assay following CLSI (formerly the NCCLS) guidelines against 16 fungal strains of clinical interest. RESULTS: In this work, fatty acid methyl esters (FAME) was able to inhibit 12 clinical strains of the pathogenic fungus Paracoccidioides brasiliensis and were also active in the bioautographic assay against Cladosporium sphaerospermum. CONCLUSIONS: FAME was a more potent antifungal than trimethoprim-sulfamethoxazole against P. brasiliensis under the experimental conditions tested.

INTRODUÇÃO: Os ácidos graxos são abundantes em óleos vegetais. Eles são conhecidos por suas propriedades antibacterianas e antifúngicas. MÉTODOS: A suscetibilidade a antifúngicos foi avaliada pelo ensaio de microdiluição em caldo de acordo com CLSI (anteriormente NCCLS) sobre 16 isolados de interesse clínico. RESULTADOS: Nesse trabalho, os ésteres metílicos de ácidos graxos (FAME) inibiram doze isolados clínicos do fungo patogênico Paracoccidioides brasiliensis, e também foi muito ativo no ensaio de bioautografia sobre o fungo Cladosporium sphaerospermum. CONCLUSÕES: FAME foi um antifúngico mais potente do que sulfametoxazol-trimetoprim contra P. brasiliensis, nas condições utilizadas no presente trabalho.