RESUMEN
Bothrops leucurus is considered as a snake of medical interest in the State of Bahia, Brazil. However, so far, there are no studies that provide a refined mapping of the composition of this venom. The aim of this work was to better understand the protein composition of B. leucurus snake venom and to isolate and biologically characterize the most abundant toxin, a basic PLA2-like. Shotgun proteomics approach identified 137 protein hits in B. leucurus venom subdivided into 19 protein families. The new basic PLA2-like toxin identified was denominated Bleu-PLA2-like, it and other proteoforms represents about 25% of the total proteins in the venom of B. leucurus and induces myotoxicity, inflammation and muscle damage. Immunoreactivity assays demonstrated that B. leucurus venom is moderately recognized by bothropic and crotalic antivenoms, and on the other hand, Bleu-PLA2-like and its proteoforms are poorly recognized. Our findings open doors for future studies in order to assess the systemic effects caused by this snake venom in order to better understand the toxinological implications of this envenomation and, consequently, to assist in the clinical treatment of victims.
Asunto(s)
Bothrops , Venenos de Crotálidos , Animales , Antivenenos/farmacología , Bothrops/metabolismo , Venenos de Crotálidos/metabolismo , Venenos de Crotálidos/toxicidad , Fosfolipasas A2/metabolismo , Venenos de Serpiente/metabolismo , Venenos de Serpiente/toxicidadRESUMEN
Bothrops leucurus is considered as a snake of medical interest in the State of Bahia, Brazil. However, so far, there are no studies that provide a refined mapping of the composition of this venom. The aim of this work was to better understand the protein composition of B. leucurus snake venom and to isolate and biologically characterize the most abundant toxin, a basic PLA2-like. Shotgun proteomics approach identified 137 protein hits in B. leucurus venom subdivided into 19 protein families. The new basic PLA2-like toxin identified was denominated Bleu-PLA2-like, it and other proteoforms represents about 25% of the total proteins in the venom of B. leucurus and induces myotoxicity, inflammation and muscle damage. Immunoreactivity assays demonstrated that B. leucurus venom is moderately recognized by bothropic and crotalic antivenoms, and on the other hand, Bleu-PLA2-like and its proteoforms are poorly recognized. Our findings open doors for future studies in order to assess the systemic effects caused by this snake venom in order to better understand the toxinological implications of this envenomation and, consequently, to assist in the clinical treatment of victims.
RESUMEN
BACKGROUND: Active packaging containing natural flavonoid has recently emerged as a result of its potential to inhibit the oxidation of foods by interacting with it and/or its surrounding environment, with the aim of counteracting oxidation reactions and extending the shelf-life of foods. The plant Spondias purpurea L. is widely found in northeastern Brazil and is known to contain bioactive flavonoids. The present study aimed to obtain a flavonoid-rich fraction from the pulp of S. purpurea L. and incorporate it into the cellulose acetate film to obtain biodegradable films with antioxidant properties. RESULTS: The fractionation in SiO2 open-column chromatography of the S. purpurea pulp crude extract furnished an antioxidant active fraction containing the flavonols quercetin 3-O-rutinoside and kaempferol 3-O-rutinoside as the major compounds. This active fraction was incorporated (10, 20 and 30 g kg-1 ) into the substance produced with the casting method for cellulose acetate films. The films produced were characterized concerning mechanical properties, water vapor permeability (WVP) and antioxidant activity. CONCLUSION: The incorporation of the active flavonoid fraction from S. purpurea in the cellulose acetate films decreases WVP and elongation at break, at the same time as increasing antioxidant activity, tensile strength and elastic modulus. Thus, the S. purpurea pulps may be an alternative as a source of antioxidants for use in cellulose acetate films. © 2020 Society of Chemical Industry.
Asunto(s)
Anacardiaceae/química , Antioxidantes/química , Celulosa/análogos & derivados , Flavonoles/química , Manipulación de Alimentos/instrumentación , Extractos Vegetales/química , Brasil , Celulosa/química , Oxidación-Reducción , Permeabilidad , Dióxido de Silicio/química , Vapor/análisis , Resistencia a la TracciónRESUMEN
This study assessed the anthelmintic activity of plant-derived compounds against gastrointestinal nematodes of goats using the egg hatch and larval motility assays. The compounds tested were saponins (digitonin and aescin) and their respective sapogenins (aglycones), hecogenin acetate and flavonoids (catechin, hesperidin, isocordoin and a mixture of isocordoin and cordoin). Additionally, cytotoxicity of active substances was analysed on Vero cell through 3-4,5-dimethylthiazol-2-yl,2,5diphenyltetrazolium bromide (MTT) and propidium iodide (PI) tests. Significant reduction on the egg hatching (P 90%). Nevertheless, higher cytotoxicity was observed in the MTT assay, with IC50 of 0.20 mg mL-1 (aescin) and 0.0074 mg mL-1 (digitonin). Aescin and digitonin have a pronounced in vitro anthelmintic effect and the glycone portion of these saponins plays an important role in this activity.
Asunto(s)
Antinematodos/farmacología , Flavonoides/farmacología , Cabras/parasitología , Nematodos/efectos de los fármacos , Oocitos/efectos de los fármacos , Saponinas/farmacología , Animales , Supervivencia Celular/efectos de los fármacos , Chlorocebus aethiops , Digitonina/farmacología , Tracto Gastrointestinal/parasitología , Concentración 50 Inhibidora , Larva/efectos de los fármacos , Compuestos de Espiro/farmacología , Esteroides/farmacología , Células VeroRESUMEN
BACKGROUND: This study was designed to assess the in vitro anthelmintic activity of the fraction containing alkaloid from Prosopis juliflora pods on goat gastrointestinal nematodes using the egg hatch assay (EHA), larval migration inhibition assay (LMIA), and larval motility assay (LMA). MATERIALS AND METHODS: The alkaloid-rich fraction (AF) - content juliprosopine as major alkaloid - was obtained from ethyl acetate extract after fractionation in Sephadex LH-20 chromatography column and its characterization were made by nuclear magnetic resonance analysis together with literature data comparison. The concentrations tested were 4.0, 2.67, 1.78, 1.19, and 0.79 mg/mL (EHA) and 4 mg/mL (LMIA and LMA). The in vitro cytotoxicity on Vero cell cultures was determined with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue tests. RESULTS: High ovicidal activity was observed with IC50 and IC90 values at 1.1 and 1.43 mg/mL for AF. On the other hand, this fraction showed low larvicidal activity and high toxic effect. CONCLUSION: Thus, P. juliflora pod alkaloid rich-fraction has ovicidal activity in vitro against goat gastrointestinal nematodes and cytotoxic in Vero cell cultures. SUMMARY: Prosopis juliflora alkaloid-rich fraction (AF) showed in vitro anthelmintic effect against gastrointestinal nematodes of goatsThe AF was more effective against eggs than third larval stage (L3) of gastrointestinal nematodesThe AF showed cytotoxicity activity on Vero cell lineThe juliprosopine was the main alkaloid found in the AF from P. juliflora pods. Abbreviations used: AF: Alkaloid-rich fraction; DMSO: Dimethyl sulfoxide; EE: Ethyl acetate extract; EHA: Egg hatch assay; IC50: Inhibitory concentration 50%; IC90: Inhibitory concentration 90%; L3: Infective larvae; LMA: Larval motility assay; LMIA: Larval migration inhibition assay; MTT: Bromide 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NMR: Nuclear magnetic resonance; PBS: Phosphate buffered saline; RPMI: Roswell Park Memorial Institute médium; TLC: Thin Layer Chromatography.
RESUMEN
This study examined the in vitro effect of the Zizyphus joazeiro bark against gastrointestinal nematodes of goats and its cytotoxicity on Vero cells. The ovicidal activity of the crude hydroethanolic extract (CE), its partitioned hexane (HE) and aqueous extract (AE) and saponins fraction (SF), including betulinic acid (BA), a biogenetic compound from this plant found in HE, were investigated using the inhibition of egg hatch assay (EHA). Thereafter, the extracts and the SF were evaluated through the larval motility assay (LMA) and larval migration inhibition assay (LMIA). The AE and SF promoted a complete inhibition of the egg hatch, and the effective concentration to inhibit 50% (EC50) values was 1.9 and 1.3mg/mL, respectively. The highest percentages of inhibition in EHA observed after treatments with CE, HE and BA corresponded to 79, 48 and 17%, respectively. The extracts and SF did not show larvicidal activity in LMA and LMIA. The AE and SF demonstrated cytotoxic effects in 3-4,5-dimethylthiazol-2-yl, 2,5diphenyltetrazolium bromide (MTT) and trypan blue tests; however, SF was more toxic (50% inhibitory concentration, IC50=0.20mg/mL). The chemical characterization of the SF was made through Proton Nuclear Magnetic Resonance ((1)H NMR) and Electrospray Ionization Mass Spectrometry (ESI-MS) analyses, which led to the identification of two saponins known as Joazeiroside B and Lotoside A. The results obtained from the research of this saponin content provide important information about the biological activity, especially the anthelmintic effect present in the plant investigated. That also suggests the types of bioactive compounds that may be responsible for this antiparasitic activity exhibited by the plant extracts.
Asunto(s)
Antihelmínticos/uso terapéutico , Enfermedades de las Cabras/tratamiento farmacológico , Parasitosis Intestinales/veterinaria , Infecciones por Nematodos/veterinaria , Extractos Vegetales/uso terapéutico , Ziziphus/química , Animales , Antihelmínticos/farmacología , Antihelmínticos/toxicidad , Chlorocebus aethiops , Heces/parasitología , Enfermedades de las Cabras/parasitología , Cabras , Concentración 50 Inhibidora , Parasitosis Intestinales/tratamiento farmacológico , Larva/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Infecciones por Nematodos/tratamiento farmacológico , Óvulo/efectos de los fármacos , Corteza de la Planta/química , Extractos Vegetales/farmacología , Extractos Vegetales/toxicidad , Saponinas/química , Saponinas/aislamiento & purificación , Saponinas/farmacología , Espectrometría de Masa por Ionización de Electrospray , Células VeroRESUMEN
The anti-nociceptive activity of an extract of Tibouchina pereirae Aubl (AETP) was investigated using two models of chemically induced pain, viz. the acetic acid-induced writhing and the formalin test, respectively, with dipyrone and indomethacin as reference drugs, respectively. In the acetic acid-induced writhing test, AETP application (100 mg/kg) caused a significant reduction of writhing produced by acetic acid. In the formalin test, AETP reduced the formalin effects significantly only in the late phase. These findings thus indicate the involvement of AETP only in peripheral antinociceptive mechanisms. In addition, AETP exhibited good antioxidant activity (EC50 approx. 15 µg/mL) in the DPPH free radical scavenging assay.