Alpha-terthienyl increases filamentous actin of Entamoeba histolytica.

This study aimed to know if alpha terthienyl (α-T) affects E. histolytica viability and to analyze its effect on the actin cytoskeleton. Trophozoites of E. histolytica HM1-IMSS were treated with α-T, then, cell viability and morphology were evaluated using tetrazolium salts and scanning electron microscopy, respectively; while actin filaments (F-actin) were stained with rhodamine-phalloidin, observed by confocal microscopy and quantified by fluorometry. Data showed that α-T inhibited cell viability of trophozoites (IC50, 19.43 µg / mL), affected the cell morphology, and increased the F-actin in a dose-dependent manner. Production of reactive oxygen species and RhoA-GTP levels remained normal in α-T-treated amebas. Two inhibitors that affect the organization of the trophozoites cytoskeleton, one that interacts directly with actin, Cytochalasin D (CD), and one that affects the Rho signaling pathway by inhibiting the downstream effector Rock, Y27632, were tested. Y27632 did not affect the increase of polymerized actin observed with α-T, this compound partially ameliorates the potent disrupting effects of CD on actin filaments. Docking results suggest that α-T could be an antagonist of CD for the same interaction zone in actin, however, more studies are needed to define the action mechanism of this compound.

A new germacranolide from Ageratina vernalis.

The aerial parts of Ageratina vernalis provided the new germacranolide 1,10-epoxydeltoidin A (3), together with the known pentacyclic triterpenoid hopane-6α,22-diol (1), and the also known germacranolides deltoidin A (2) and 15-hydroxydeltoidin A (4). In addition, pTsOH catalyzed cyclization of 2 afforded the new guaianolide 5. The absolute configuration of 2, 4, and 5 was assigned by vibrational circular dichroism spectroscopy, while the complete 1H and 13C NMR data assignments of 2-5 followed from 1 D- and 2 D-NMR experiments.

Chemical Characterization of Plant Extracts and Evaluation of their Nematicidal and Phytotoxic Potential.

Nacobbus aberrans ranks among the "top ten" plant-parasitic nematodes of phytosanitary importance. It causes significant losses in commercial interest crops in America and is a potential risk in the European Union. The nematicidal and phytotoxic activities of seven plant extracts against N. aberrans and Solanum lycopersicum were evaluated in vitro, respectively. The chemical nature of three nematicidal extracts (EC50,48h ≤ 113 µg mL-1) was studied through NMR analysis. Plant extracts showed nematicidal activity on second-stage juveniles (J2): (≥87%) at 1000 µg mL-1 after 72 h, and their EC50 values were 71.4-468.1 and 31.5-299.8 µg mL-1 after 24 and 48 h, respectively. Extracts with the best nematicidal potential (EC50,48h < 113 µg mL-1) were those from Adenophyllum aurantium, Alloispermum integrifolium, and Tournefortia densiflora, which inhibited L. esculentum seed growth by 100% at 20 µg mL-1. Stigmasterol (1), ß-sitosterol (2), and α-terthienyl (3) were identified from A. aurantium, while 1, 2, lutein (4), centaurin (5), patuletin-7-ß-O-glucoside (6), pendulin (7), and penduletin (8) were identified from A. integrifolium. From T. densiflora extract, allantoin (9), 9-O-angeloyl-retronecine (10), and its N-oxide (11) were identified. The present research is the first to report the effect of T. densiflora, A. integrifolium, and A. aurantium against N. aberrans and chemically characterized nematicidal extracts that may provide alternative sources of botanical nematicides.

Characterization of pectin extracted under mild conditions from tempesquistle (Sideroxylon palmeri) fruit at two maturity stages.

BACKGROUND: le tree (Sideroxylon palmeri) belongs to family Sapotaceae, and its fruits contain hydrophilic and hydrophobic gums. Aim of this study was to develop an extraction method for pectin, a hydrophilic gum, from green and ripe tempesquistle fruit. The extraction method was gentle to minimize potential structural changes in pectin, a maceration was used, and then pectin precipitation was induced with acetone at room temperature. Pectin characterization was carried out in two steps. The first step consisted of qualitative tests of Molisch, Fehling and Lugol. The second step consisted of quantitative tests to determine esterification degree (ED) by ATR-FTIR and D-galacturonic acid (D-GalA) content with a colorimetric assay. ATR-FTIR spectroscopic method revealed that green and ripe tempesquistle fruit have an ED of 0% and 30.4%, respectively. Both fruit have a low ED pectin. The content of D-GalA in green and ripe pectins was 18.8 ±2.7% and 20.2 ±0.6%, respectively. The yield obtained in green and ripe fruit samples was 1.6 ±0.2% and 3.0 ±0.1%, respectively. The extraction method allowed two types of pectins obtained in function of maturity stage of tempesquistle fruit distinguishable by. METHODS: istle tree (Sideroxylon palmeri) belongs to family Sapotaceae, and its fruits contain hydrophilic and hydrophobic gums. Aim of this study was to develop an extraction method for pectin, a hydrophilic gum, from green and ripe tempesquistle fruit. The extraction method was gentle to minimize potential structural changes in pectin, a maceration was used, and then pectin precipitation was induced with acetone at room temperature. Pectin characterization was carried out in two steps. The first step consisted of qualitative tests of Molisch, Fehling and Lugol. The second step consisted of quantitative tests to determine esterification degree (ED) by ATR-FTIR and D-galacturonic acid (D-GalA) content with a colorimetric assay. ATR-FTIR spectroscopic method revealed that green and ripe tempesquistle fruit have an ED of 0% and 30.4%, respectively. Both fruit have a low ED pectin. The content of D-GalA in green and ripe pectins was 18.8 ±2.7% and 20.2 ±0.6%, respectively. The yield obtained in green and ripe fruit samples was 1.6 ±0.2% and 3.0 ±0.1%, respectively. The extraction method allowed two types of pectins obtained in function of maturity stage of tempesquistle fruit distinguishable by ATR. RESULTS: istle tree (Sideroxylon palmeri) belongs to family Sapotaceae, and its fruits contain hydrophilic and hydrophobic gums. Aim of this study was to develop an extraction method for pectin, a hydrophilic gum, from green and ripe tempesquistle fruit. The extraction method was gentle to minimize potential structural changes in pectin, a maceration was used, and then pectin precipitation was induced with acetone at room temperature. Pectin characterization was carried out in two steps. The first step consisted of qualitative tests of Molisch, Fehling and Lugol. The second step consisted of quantitative tests to determine esterification degree (ED) by ATR-FTIR and D-galacturonic acid (D-GalA) content with a colorimetric assay. ATR-FTIR spectroscopic method revealed that green and ripe tempesquistle fruit have an ED of 0% and 30.4%, respectively. Both fruit have a low ED pectin. The content of D-GalA in green and ripe pectins was 18.8 ±2.7% and 20.2 ±0.6%, respectively. The yield obtained in green and ripe fruit samples was 1.6 ±0.2% and 3.0 ±0.1%, respectively. The extraction method allowed two types of pectins obtained in function of maturity stage of tempesquistle fruit distinguishable by ATR-FTIR.

Insecticidal and Nematicidal Contributions of Mexican Flora in the Search for Safer Biopesticides.

Plant metabolites have been used for many years to control pests in animals and to protect crops. Here, we reviewed the available literature, looking for the species of Mexican flora for which extracts and metabolites have shown activity against pest insects and parasitic nematodes of agricultural importance, as well as against nematodes that parasitize domestic cattle. From 1996 to 2018, the search for novel and eco-friendly biopesticides has resulted in the identification of 114 species belonging to 36 botanical families of Mexican plants with reported biological effects on 20 insect species and seven nematode species. Most plant species with detected pesticide properties belong to the families Asteraceae, Fabaceae, and Lamiaceae. Eighty-six metabolites have been identified as pesticidal active principles, and most have been terpenoids. Therefore, the continuation and intensification of this area of research is very important to contribute to the generation of new products that will provide alternatives to conventional pesticide agents. In addition, future studies will contribute to the recognition and dissemination of the importance of propagating plant species for their conservation and sustainable use.

Productivity and Antioxidant Activity of Wild, Reconstituted, and Hybrid Strains of the Pink Oyster Mushroom, Pleurotus djamor (Agaricomycetes), from Mexico.

The genus Pleurotus is the third most commonly produced edible fungi in the world. In addition, species of genus Pleurotus have functional properties such as anticancer, antiviral, antimicrobial, anti-inflammatory, and antioxidant activities, which are mainly attributed to phenolic compounds. For these reasons, this study evaluated the productivity and antioxidant activity (AA) of 2 wild strains (white and pink), 2 reconstituted strains (called "BB" and "RR"), and 4 hybrid strains (H1, H2, H3, and H4) of P. djamor from monokaryotic components (neohaplonts). The results showed that the white wild-type strain and the reconstituted strains exhibited the best production potential, expressed as biological efficiency and mycelial growth rate. The carpophores of hybrid strains H1 and H3 had the greatest AA, as evaluated with DPPH radical scavenging and reducing power assays, respectively. The H3 strain had the highest total phenol (TP) content. Pearson correlations led us to conclude that the mycelial growth rate has a regular inverse correlation with TP and a regular direct correlation with AA of methanolic extracts from carpophores and myce-lia. This is, to our knowledge, the first report in the literature about the effect of Pleurotus strain hybridization through a chemical de-dikaryotization process on TP content.

Natural antioxidant extracts as food preservatives.

The food industry is becoming more specialized and processing methods are continuously being developed to meet consumer needs. Consumers demand products that are safe and preferably free of synthetic additives. These additives are associated with health effects, in most cases without reasonable justification. Consequently, consumers are looking for clearly labelled products that guarantee the absence of synthetic additives. This has led to the need to search for natural additives, which the food industry claims arenatural antioxidant preservatives. The sources of natural antioxidants can be extremely varied, because practically all plants contain antioxidants that allow them to protect themselves from solar radiation and pests, as well as to regulate the production of chemical energy. However, the best alternatives for the food industry are fruits and spices, because they are already foods themselves. This article will describe fruits and spices considered as important sources of phenolic antioxidants. The main medicinal properties are related to phenolic compounds and their uses as additives, depending on their chemical structure.

EVALUATION OF ANTICANCER POTENTIAL OF EIGHT VEGETAL SPECIES FROM THE STATE OF OAXACA.

BACKGROUND: Eight plant species from Oaxaca, some of them used in traditional medicine, were subjected to screening of several biological activities to provide data regarding their anticancer potential, although no scientific information is available about their pharmacological effects. MATERIALS AND METHODS: Methanol extracts from stems or roots of the eight plants were tested for antioxidant activity by the DPPH- method. Antimicrobial activity was determined using the agar diffusion method and the minimal inhibitory concentration (MIC) was obtained by broth dilution method. Antitopoisomerase activity was assessed using mutant strains of Saccharomyces cerevisiae JN362a, JN394, JN394t-1, JN394t2.4 and JN394t2-5. The mutagenic activity was evaluated using the Ames test (Salmonella typhimurium TA1535). RESULTS: No extract showed significant antioxidant activity. The best antimicrobial activity was observed for Salpianthus arenarius (MIC 56.25 µg/mL) and Lantana achyranthifolia (MIC 78.12 µg/mL) against Staphylococcus aureus. Extracts of Acalypha cuspidata, Alloispermum integrifolium and L. achyranthifolia stems showed antitopoisomerase II activity with JN394t-1 growth of -30.88±0.0%, -38.11±4.95%, and -70.97±12.02% respectively. Galium mexicanum stem extract showed antitopoisomerase I activity with growth of 35.31±6.36% on the same mutant strain. All plant extracts were non-mutagenic. Fractionation of A. cuspidata extract led to identification of two subfractions with antitopoisomerase I and II activity at 154µg/mL (Positive controls 50 and 100µg/mL). CONCLUSION: Methanol extracts of A. cuspidata, A. integrifolium, G. mexicanum, and L. achyranthifolia stems showed antitopoisomerase and non-mutagenic activities, and consequently could be promising as a source of anticancer drugs.

Antiamoebic Activity of Adenophyllum aurantium (L.) Strother and Its Effect on the Actin Cytoskeleton of Entamoeba histolytica.

In Mexico, the Adenophyllum aurantium (L.) Strother plant is consumed as an infusion to treat intestinal diseases such as amoebiasis, which is an endemic health problem in Mexico and other countries. However, the effect of A. aurantium on Entamoeba histolytica, the causative agent of amoebiasis, is unknown. An aerial part methanolic extract (AaMeA), a root methanolic extract (AaMeR) and a root ethyl acetate extract (AaEaR) were tested on E. histolytica trophozoites. AaMeA and AaMeR did not show antiproliferative activity; however, AaEaR exhibited an in vitro GI50 of 230 µg/ml, and it was able to inhibit the differentiation of Entamoeba invadens trophozoites into cysts. The intraperitoneal administration of AaEaR (2.5 or 5 mg) to hamsters that were infected with E. histolytica inhibited the development of amoebic liver abscesses in 48.5 or 89.0% of the animals, respectively. Adhesion to fibronectin and erythrophagocytosis were 28.7 and 37.5% inhibited by AaEaR, respectively. An ultrastructure analysis of AaEaR-treated trophozoites shows a decrease in the number of vacuoles but no apparent cell damage. Moreover, this extract affected the actin cytoskeleton structuration, and it prevented the formation of contractile rings by mechanism(s) that were independent of reactive oxygen species and RhoA activation pathways. (13)C NMR data showed that the major compounds in the AaEaR extract are thiophenes. Our results suggest that AaEaR may be effective in treatments against amoebiasis, nevertheless, detailed toxicity studies on thiophenes, contained in AaEaR, are required to avoid misuse of this vegetal species.

Effect of crude plant extracts from some Oaxacan flora on two deleterious fungal phytopathogens and extract compatibility with a biofertilizer strain.

The antimicrobial activity of 12 plant extracts was tested against the phytopathogens Alternaria alternata and Fusarium solani. In addition, the compatibility of the extracts toward Bacillus liqueniformis, a biofertilizer and a non-target microorganism, was assessed. Plants tested belong to the Euphorbiaceae, Asteraceae, Crassulaceae, Rubiaceae, Convolvulaceae, Verbenaceae, Orchidaceae, Nyctaginaceae, Boraginaceae, and Tiliaceae families and were collected in the State of Oaxaca. The antifungal activity of the plant extracts (50-100 mg/mL) against A. alternata and F. solani, was determined by measuring the mycelium radial growth and obtaining the minimum inhibitory concentration (MIC) of fungal growth. In addition, with the aim of finding plant extracts which are compatible with a B. licheniformis biofertilizer strain and to test the non-toxic nature of the treatments, the toxicity of the extracts toward this strain was evaluated using the agar diffusion method. Azoxystrobin (12 µg) and chloramphenicol (30 µg) were used as positive controls for the pathogens and for the non-target bacteria, respectively. Plant extracts inhibited fungal growth in the ranges of 0.76-56.17% against F. solani and 2.02-69.07% against A. alternata. The extracts of Acalypha subviscida, Ipomoea murucoides, Tournefortia densiflora and Lantana achyranthifolia showed MIC values between 5.77-12.5 mg/mL for at least one of the fungal species. The best treatment, Adenophyllum aurantium, exhibited a maximum inhibition for both F. solani (56.17%, MIC = 7.78 mg/mL) and A. alternata (68.64% MIC = 7.78 mg/mL), and resulted innocuous toward B. licheniformis. Therefore, this plant has an outstanding potential for the agroecological control of fungal phytopathogens in industrial crops.

Complete 1H NMR assignments of pyrrolizidine alkaloids and a new eudesmanoid from Senecio polypodioides.

Chemical investigation of the aerial parts of Senecio polypodioides lead to the isolation of the new eudesmanoid 1ß-angeloyloxyeudesm-7-ene-4ß,9α-diol (1) and the known dirhamnosyl flavonoid lespidin (3), while from roots, the known 7ß-angeloyloxy-1-methylene-8α-pyrrolizidine (5) and sarracine N-oxide (6), as well as the new neosarracine N-oxide (8), were obtained. The structure of 1 and 8 was elucidated by spectral means. Complete assignments of the (1)H NMR data for 5, 6, sarracine (7), and 8 were made using one-dimensional and two-dimensional NMR experiments and by application of the iterative full spin analysis of the PERCH NMR software.

Bioactive saponins from Microsechium helleri and Sicyos bulbosus.

Eleven oleanane-type saponins (1-11) have been isolated from Microsechium helleri and Sicyos bulbosus roots and were evaluated for their antifeedant, nematicidal and phytotoxic activities. Saponins {3-O-ß-D-glucopyranosyl (1→3)-ß-D-glucopyranosyl-2ß,3ß,16α,23-tetrahydroxyolean-12-en-28-oic acid 28-O-α-L-rhamnopyranosyl-(1→3)-ß-D-xylopyranosyl-(1→4)-[ß-D-xylopyranosyl-(1→3)]-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranoside} (1), and {3-O-ß-D-glucopyranosyl-2ß,3ß,16α,23-tetrahydroxyolean-12-en-28-oic acid 28-O-α-L-rhamnopyranosyl-(1→3)-ß-D-xylopyranosyl-(1→4)-[ß-D-xylopyranosyl-(1→3)]-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranoside} (2) were also isolated from M. helleri roots together with the two known compounds 3 and 4. Seven known structurally related saponins (5-11) were isolated from S. bulbosus roots. The structures of these compounds were established as bayogenin and polygalacic glycosides using one- and two-dimensional NMR spectroscopy and mass spectrometry. Compounds 7, 10, bayogenin (12) and polygalacic acid (13) showed significant (p<0.05) postingestive effects on Spodoptera littoralis larvae, compounds 5-11 and 12 showed variable nematicidal effects on Meloydogyne javanica and all tested saponins had variable phytotoxic effects on several plant species (Lycopersicum esculentum, Lolium perenne and Lactuca sativa). These are promising results in the search for natural pesticides from the Cucurbitaceae family.

New saponins from Sechium mexicanum.

The chemical study of Sechium mexicanum roots led to the isolation of the two new saponins {3-O-beta-D-glucopyranosyl (1 --> 3)-beta-D-glucopyranosyl-2beta,3beta,16alpha,23-tetrahydroxyolean-12-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L-arabinopyranoside} (1) and {3-O-beta-D-glucopyranosyl (1 --> 3)-beta-D-glucopyranosyl-2beta,3beta,16alpha,23-tetrahydroxyolean-12-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-[beta-D-apiosyl-(1 --> 3)]-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L-arabinopyranoside} (2), together with the known compounds {3-O-beta-D-glucopyranosyl-(1 --> 3)-beta-D-glucopyranosyl-2beta,3beta,6beta,16alpha,23-pentahydroxyolean-12-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)-alpha-L-rhamnopyranosyl-(1 --> 2)-alpha-L-arabinopyranoside} (3), tacacosides A(1) (4) and B(3) (5). The structures of saponins 1 and 2 were elucidated using a combination of (1)H and (13)C 1D-NMR, COSY, TOCSY, gHMBC and gHSQC 2D-NMR, and FABMS of the natural compounds and their peracetylated derivates, as well as by chemical degradation. Compounds 1-3 are the first examples of saponins containing polygalacic and 16-hydroxyprotobasic acids found in the genus Sechium, while 4 and 5, which had been characterized partially by NMR, are now characterized in detail.

Profiling of the resin glycoside content of Mexican jalap roots with purgative activity.

Mexican Jalap roots, a prehispanic medicinal plant complex still considered to be a useful laxative, can be found as an ingredient in some over-the-counter products sold by herbalists in contemporary Mexico. The drug is prepared from the dried roots of several morning glories, all of which have been identified as members of the genus Ipomoea. Analysis of several commercial samples was assessed by generating HPLC and 13C NMR spectroscopic profiles of the glycosidic acids obtained through saponification of the resin glycoside contents. These profiles distinguish the three Mexican jalaps currently in frequent use and can serve as analytical tools for the authentication and quality control of these purgative herbal drugs. Ipomoea purga, the authentic "jalap root", yielded two new hexasaccharides of convolvulinic and jalapinolic acids, purgic acids A (1) and B (2), respectively. Scammonic acid A (3), a tetrasaccharide, was produced from Ipomoea orizabensis, the Mexican scammony or false jalap. Operculinic acid B (4), a pentasaccharide, was identified in Ipomoea stans. Semipreparative HPLC was performed to obtain pure samples of new compounds 1 and 2 in sufficient quantity to elucidate their structure by high-field NMR spectroscopy. Purgic acid A (1) was identified as (11S)-hydroxytetradecanoic acid 11-O-beta-D-quinovopyranosyl-(1-->2)-O-beta-D-glucopyranosyl-(1-->3)-O-[beta-D-fucopyranosyl-(1-->4)]-O-alpha-L-rhamnopyranosyl-(1-->2)-O-beta-D-glucopyranosyl-(1-->2)-O-beta-D-quinovopyranoside, while purgic acid B (2) was characterized with (11S)-hydroxyhexadecanoic acid as its aglycon but having the same glycosidation sequence in the oligosaccharide core.