Galanthamine and other Amaryllidaceae related alkaloids are inhibitors of α7, α4β2 and α3β4 nicotinic acetylcholine receptors

Abstract Galanthamine is an Amaryllidaceae-derived acetylcholinesterase inhibitor used to treat memory impairment in Alzheimer's disease and vascular dementia. There is evidence that galanthamine, in addition to its effects on acetylcholinesterase, may enhance or inhibit brain nicotinic acetylcholine receptors, which could increase or decrease the therapeutic efficacy of galanthamine, respectively. Here, we evaluated the effects of galanthamine and two others Amaryllidaceae acetylcholinesterase inhibitors (haemanthamine and tazettine) analyzed by gas chromatography-mass spectrometry and identified by comparing their mass fragmentation patterns with literature and database NIST vs.2.0 on the agonist responses of brain nicotinic acetylcholine receptors α7, α3β4, (α4)2(β2)3 and (α4)3(β2)2. Using nicotinic acetylcholine receptors expressed heterologously in Xenopus oocytes, in conjunction with two-electrode voltage clamping, we found that galanthamine inhibits the function of nicotinic acetylcholine receptors assayed through a mix competitive and non-competitevely. Nicotinic acetylcholine receptor α7 were significantly more sensitive to inhibition (17 ± 0.6 µM) than the heteromeric receptor, α3β4 (90 ± 3.4 µM). Neither haemanthamine nor tazettine were more potent than galanthamine.

Differential formononetin content in cultivars and experimental lines of red clover (Trifolium pratense L. ) plants affect the feeding behaviour of Hylastinus obscurus (Coleoptera: Curculionidae) / Contenido diferencial de formononetina en cultivares y líneas experimentales de plantas de trébol rosado (Trifolium pratense L. ) afecta la conducta alimenticia de Hylastinus obscurus (Coleoptera: Curculionidae)

Red clover (Trifolium pratense L.) is a perennial plant widely used as a forage resource for several animals. This plant is the exclusive host of Hylastinus obscurus (Marsham) which causes irreparable damages to the root system affecting their persistence. It has been reported that the presence of the isoflavonoid formononetin in roots of red clover could act as an antifeedant on H. obscurus. There are not studies related to the formononetin content in red clover roots to the antifeedant effect elicited by experimental lines and cultivar of red clover. Six red clover genotypes were investigated in both formononetin content and their respective antifeedant action. The results showed to Sabtoron High and Superqueli-INIA with both the highest formononetin content in red clover roots and antifeedant effect, allowing to suggest that this secondary metabolites could be used as a chemical factor for red clover plants. Moreover, a rapid methodology for searching red clover genotypes with high formononetin content is reported.

El trébol rosado (Trifolium pratense L.) es una planta perenne ampliamente utilizada como fuente de forraje de variados animales. Esta planta es el exclusivo hospedero de Hylastinus obscurus (Marsham) el cual causa irreparables daños al sistema radical afectando seriamente su persistencia. Se ha reportado que la presencia del isoflavonoide formononetina en raíces del trébol rosado podría actuar como antialimentario sobre H. obscurus. Actualmente no existen estudios que relacionen el contenido de formononetina en raíces de trébol rosado con el efecto antialimentario elicitado por líneas experimentales y cultivares de trébol rosado. Seis genotipos de esta leguminosa fueron evaluados en cuanto a su contenido de formononetina y actividad antialimentaria. Los resultados mostraron que los cultivares Sabtoron High y Superqueli-INIA presentaron altos niveles de formononetina en sus raíces y efecto antialimentario sobre H. obscurus, lo que permite sugerir que este metabolito secundario podría ser usado como factor químico para incrementar la persistencia de plantas de trébol rosado. Además, se informa una metodología rápida para la búsqueda de genotipos con altos contenidos de formononetina.

Rhodolirium andicola: a new renewable source of alkaloids with acetylcholinesterase inhibitory activity, a study from nature to molecular docking

ABSTRACT Acetylcholinesterase is an important target for control of neurodegenerative diseases causing cholinergic signaling deficit. Traditionally, galanthamine has been used as an Amaryllidaceae-derived acetylcholinesterase inhibitor, although new Amaryllidaceae plants could serve as source for better acetylcholinesterase inhibitors. Therefore, the objective of this study was to characterize the alkaloid composition from bulbs of Rhodolirium andicola (Poepp.) Traub, a native Chilean Amaryllidaceae specie, and assess their inhibitory activity on acetylcholinesterase by in vitro and in silico methodologies. Alkaloidal extracts from R. andicola exhibited an inhibitory activity with IC50 values between 11.25 ± 0.04 and 57.78 ± 1.92 µg/ml that included isolated alkaloid, galanthamine (2.3 ± 0.18 µg/ml), Additionally, 12 alkaloids were detected using gas chromatography-mass spectrometry and identified by comparing their mass fragmentation patterns with literature and database NIST vs.2.0. To better understand the bioactivity of isolated compounds and alkaloidal extracts against acetylcholinesterase, a molecular docking approach was performed. Results suggested that alkaloids such as lycoramine, norpluvine diacetate and 6α-deoxy-tazettine expand the list of potential acetylcholinesterase inhibitors to not only galanthamine. The role of R. andicola as a source for acetylcholinesterase inhibitors is further discussed in this study.

Identification of isoquinoline alkaloids from Berberis microphylla by HPLC ESI-MS/MS / Identificación de alcaloides isoquinolínicos en Berberis microphylla G. Forst mediante CLAE IES-MS/MS

Berberis microphylla (G. Forst) is a native plant growing in Patagonia. In recent years Patagonia Berberis are becoming important due to their interesting biological properties related to their alkaloids content. The aim of this study was determine the distribution and proportion of isoquinoline alkaloids in leaves, stems and roots of B. microphylla collected in two different climatic zones from Chilean Patagonia. Using by HPLC ESI-MS/MS isocorydine, jatrorrhizine, palmatine, reticuline, scoulerine, tetrahydroberberine and thalifendine were detected for the first time in this specie, and the presence of allocryptopine, berberine, calafatine and protopine, previously isolated in B. microphylla was corroborated. The alkaloids profile showed differences of compounds in samples collected in two climatic zones, where more compounds were detected in plants from Lago Deseado than Cerro Sombrero. Furthermore, a greater number of alkaloids were found in stem and root extracts and berberine and thalifendine were detected in higher proportion in these structures.

Berberis microphylla (G. Forst) es un arbusto nativo que crece en la Patagonia. Actualmente, esta planta ha sido foco de estudio dada las propiedades biológicas que presenta, atribuidas principalmente al contenido de alcaloides. El objetivo de este estudio fue determinar la distribución y proporción de alcaloides isoquinolínicos en hojas, tallos y raíces de B. microphylla colectadas en dos zonas climáticas de la Patagonia chilena. Mediante CLAE IES-MS/MS se informa por primera vez la presencia de isocoridina jatrorrizina, palmatina, reticulina, escoulerina, tetrahidroberberina y talifendina en esta especie y se confirma la presencia de allocriptopina, berberina, calafatina y protopina, identificados previamente en B. microphylla. El perfil de alcaloides mostró diferencias en la presencia de compuestos en las muestras colectas en las dos zonas climáticas, observándose un mayor número de compuestos en plantas provenientes de Lago Deseado. Además, un mayor número de compuestos se identificó en extractos de tallos y raíces donde berberina y talifendina fueron detectados en mayor proporción.

Novel antimicrobial activity of a dichloromethane extract obtained from red seaweed Ceramium rubrum (Hudson) (Rhodophyta: Florideophyceae) against Yersinia ruckeri and Saprolegnia parasitica, agents that cause diseases in salmonids

Background Enteric red mouth disease and Saprolegniasis, which are caused by the bacteria Yersinia ruckeri and the oomycete Saprolegnia parasitica, respectively, are important illnesses that affect salmonid farming. Sanitary problems in farms are addressed by the prevention of disease outbreaks or by the treatment of diseases with chemicals. Environmental and governmental restrictions, toxicity and high treatment costs limit the use of drugs. Marine organisms, such as algae, sponges and corals, have developed an antimicrobial defense strategy based on the production of bioactive metabolites. Among these organisms, seaweeds offer a particularly rich source of potential new drugs. Hence, many pharmacologically active substances have been isolated from seaweeds. In the Ceramium genus, Ceramium rubrum has been emphasized by several authors for its antimicrobial properties. Based on this background, the present study focused on the antimicrobial activity of a lipophilic extract of C. rubrum on Y. ruckeri and S. parasitica. Results The alga, collected from the Pacific coast of Chile, underwent an ethanol extraction, and the concentrated extract was partitioned between water and dichloromethane. From the dichloromethane extract, fatty acids, fatty acid esters, one hydrocarbon and phytol were identified by Gas Chromatography-Mass Spectrometry (GC/MS) analysis. The antimicrobial study showed that the whole extract was more active than the individual components, which suggests a strong synergistic effect among the components. Conclusions These results may constitute a basis for promising future applied research that could investigate the use of C. rubrum seaweed as a source of antimicrobial compounds against fish pathogens.

Chemical composition and antibacterial activity of Laureliopsis philippiana (Looser) essential oil / Composición química y actividad antibacteriana del aceite esencial de Laureliopsis philippiana (Looser)

Laureliopsis philippiana (Looser) is native evergreen specie from Chile and Argentina used in traditional medicine. In this study, chemical composition as well as its in vitro antibacterial activity against Escherichia coli, Enterobacter aerogenes, Staphylococcus epidermidis and Staphylococcus aureus of essential oil from leaves of this species was determinated. Chemical analysis by GC-MS resulted in the identification of 19 compounds representing 98.8 percent onoterpenes; linalool (32.3 percent) and eucalyptol (37.4 percent) were the main constituents. To evaluate the antibacterial activity disc diffusion method and broth dilution method were used. The essential oil exhibited inhibitory activity against Gram (-) and Gram (+) bacteria, whereas similar activity to essential oil was showed for linalool against E. aerogenes and S. epidermidis whereas linalool alone, achieves an inhibitory effect against E. aerogenes and S. epidermidis comparable to the essential oil.

Laureliopsis philippiana (Looser) es una especie siempre verde nativa de Chile y Argentina usado en medicina tradicional. En este estudio se determinó tanto la composición química del aceite esencial obtenido a partir de hojas de esta especie, así como su actividad antibacterial in vitro contra Escherichia coli, Enterobacter aerogenes, Staphylococcus epidermidis and Staphylococcus aureus. El análisis químico por GC-MS permitió la identificación de 19 compuestos, representando el 98,8 por ciento de la composición del aceite. Monoterpenos oxigenados, eucaliptol y linalol fueron los mayores constituyentes del aceite con un 37,4 por ciento y 32,3 por ciento respectivamente. Para evaluar la actividad antibacteriana se utilizaron los métodos de difusión en agar y dilución en caldo. El aceite esencial muestra actividad inhibitoria contra las bacterias Gram (-) y Gram (+) evaluadas, mientras que linalol por si solo logra un efecto inhibitorio comparable con el aceite esencial contra E. aerogenes y S. epidermidis mientras que el linalol por si solo, logra un efecto inhibitorio contra E. aerogenes y S. epidermidis comparable al del aceite esencial.

Differences in the fly-load of Haematobia irritans (Diptera: Muscidae) on cattle is modified by endophyte infection of pastures

Background: The horn fly, Haematobia irritans, is an obligate bloodsucking ectoparasite of pastured cattle and is a major pest of livestock production in North and South America and Europe. In this study, we investigated the potential to use cattle pastures, infected with non-toxic, "friendly" fungal-endophyte-infected (E+) tall fescue, Festuca arundinacea Schreb., as a strategy for reducing horn fly loads in cattle, and to evaluate the possible bioinsecticide effect on horn fly larvae. Results: When cattle grazed in E+ tall fescue, a decrease in fly-load was observed, compared with other pastures (endophyte-free (E-) pastures). The infestation of horn fly load decreased according to an increase in the percentage of endophyte present in the different pastures (0 to 100%). Moreover, two groups of animals with significant differences in the fly-load (high and low fly-load) in the same herd were observed (P < 0.05). Additionally, it was possible to determine a bioinsecticide effect of cattle dung, upon horn fly larvae (80%), from animals fed E+ tall fescue. Conclusions: These results constitute the first report on the potential for exploiting pasture management for controlling 1) horn fly-loads on cattle and 2) the normal development of horn fly larvae. In conclusion, this information provides preliminary understanding of the role of cattle pasture diet management for controlling horn fliesas part of an integrated pest management strategy for this major pest of farmed livestock.