RESUMO
Physic nut Jatropha curcas cake/meal obtained after oil extraction has a high protein content, however, the presence of antinutrients (trypsin inhibitor, lectin and phytate) and toxic compounds (phorbol esters) limit their use as an alternative feedstuff. Thus, the detoxification process in cake/meal is necessary to allow their inclusion in fish diets. The present study aimed to evaluate the effects of solvent and extrusion-treated jatropha cake (SETJC) in Nile tilapia (Oreochromis niloticus) diets on growth, body composition, nutrient utilization, metabolic and hematological responses, and digestibility of experimental diets. Five experimental diets were formulated to be isonitrogenous (28.50% digestible protein) and isoenergetic (13.39 MJ/kg digestible energy) with graded levels of SETJC (0, 3, 6, 9, and 12%). The experimental design was completely randomized with five treatments and four replicates. The detoxification treatments reduced the phorbol esters (PE) of jatropha cake by 96% (0.58 mg/g of PE before and 0.023 mg/g of PE after treatments). Increased levels of SETJC depressed growth, feed efficiency, and protein digestibility. A similar trend was observed for hematological and biochemistry parameters. Aspartate and alanine aminotransferase, as well as phosphorus and magnesium concentrations in the fillets, increased at the highest levels of SETJC. Thus, the data of the present study suggests that the residual content, different structural forms of phorbol ester and its biological activity, as well as some antinutritional factors, can influence negatively the growth, metabolism and digestibility of experimental diets for Nile tilapia.
Assuntos
Ciclídeos , Jatropha , Animais , Jatropha/química , Jatropha/metabolismo , Ração Animal/análise , Solventes/análise , Dieta/veterinária , Ésteres de Forbol/farmacologia , Ésteres de Forbol/análise , Ésteres de Forbol/metabolismo , Sementes/química , Sementes/metabolismoAssuntos
Abrasão Química , Croton , Óleo de Cróton , Glicolatos , Humanos , Fenol , Óleo de Gergelim , Ácido TricloroacéticoRESUMO
BACKGROUND: Casbene synthase (CS) is responsible for the first committed step in the biosynthesis of phorbol esters (PE) in the Euphorbiaceae. PE are abundant in the seeds of the biofuel crop Jatropha curcas and its toxicity precludes the use of the protein-rich cake obtained after oil extraction as an animal feed and the toxicity of the fumes derived from burning PE containing biofuel is also a matter of concern. This toxicity is a major hindrance to exploit the potential of this crop as a source of raw material to produce biodiesel. For this reason, the current research on J. curcas is mainly focused on the understanding of the biosynthesis and site of synthesis of PE, as an avenue for the development of genotypes unable to synthesize PE in its seeds. RESULTS: Here, we present targeted proteomics assays (SRM and PRM) to detect and quantify CS in leaves, endosperm, and roots of two J. curcas genotypes with contrasting levels of PE. These assays were based on the use of reference isotopic labeled synthetic peptides (ILSP) predicted from 12 gene models of CS from the J. curcas genome. CONCLUSION: Our targeted proteomics methods were able to detect and quantify, for the first time, CS gene products and demonstrate the distribution of CS isoforms only in roots from J. curcas genotypes with a high and low concentration of PE. These methods can be expanded to monitor CS, at the protein level, in different tissues and genotypes of J. curcas.
RESUMO
Today, a great interest in Jatropha-based products exists worldwide, mainly for the production of biofuel.However, the oil obtained from this plant is known to be toxic due to contained curcins andphorbol esters. Bioassays, including plant cytogenetic assays based on cell cycle observation, are useful for determining the toxicity of J. curcas oil. Hence, the aim of this study was to describe the mechanism of action of J. curcas oil by cell cycle analysis using Lactuca sativa as plant testing model. A decrease in root growth was observed, closely related to the reduction in mitotic index, along with an increase in condensed nuclei. J. curcas chemicals act both as aneugenic agents, leading to the formation of lagged, sticky chromosomes and c-metaphase cells, as well as clastogenic agents, inducing the formation of chromosome bridges and fragments. The cytotoxicity and genotoxicity of phorbol esters and other chemical components of J. curcas oil was determined and discussed.
Um grande interesse mundial existe em produtos à base de pinhão manso, principalmente para a produção de biocombustíveis. No entanto, o óleo obtido a partir desta planta é conhecidamente tóxico por conter curcina e ésteres de forbol. Bioensaios, incluindo ensaios citogenéticos em plantas-modelo com base na observação do ciclo celular, são úteis para determinar a toxicidade do óleo de J. curcas. Assim, o objetivo deste estudo foi descrever o mecanismo de ação do óleo de J. curcas por análise do ciclo celular usando Lactuca sativa como modelo de teste em plantas. Foi observada uma redução no crescimento das raízes, intimamente relacionada com a redução do índice mitótico e com um aumento de núcleos condensados. Os constituintes químicos de J. curcas atuam simultaneamente como agentes aneugênicos, levando à formação de cromossomos perdidos e pegajosos e células em c-metáfase, bem como agentes clastogênicos, induzindo a formação de pontes e fragmentos cromossômicos. A citotoxicidade e genotoxicidade do éster de forbol e outros componentes químicos do óleo de J. curcas foram determinados e discutidos.
Assuntos
Ciclo Celular , Aneugênicos , Jatropha , Toxicidade , Índice MitóticoRESUMO
In this study, a molecularly imprinted polymer-coated probe electrospray ionization mass spectrometry (MIPCPESI-MS) method was developed for detection of phorbol esters (PEs) and deoxyphorbol metabolites in Jatropha curcas leaves. Such an approach was established by sticking on a metallic needle a molecularly imprinted polymer to particularly design a MIP-coated probe for selective sampling and ionization of PEs and deoxyphorbol metabolites. By a subsequent application of a high voltage and methanol, as spray solvent, ESI was generated for direct and rapid analysis under ambient and open-air conditions. MIP-coated probe exhibited a high sampling capacity of the PEs and its metabolites in methanolic extracts of J. curcas leaves compared with the non-imprinted polymer (NIP)-coated probe. MIPCPESI-MS allowed the detection of phorbol 12,13-diacetate (PDA) from J. curcas leaves with minimal sample preparation, and with detection limit and quantification reaching 0.28 µg/mL and 0.92 µg/mL, respectively. Also, good linearity was obtained with R2 > 0.99 and precision and accuracy values between 4.06-13.49% and - 1.60 to - 15.26%, respectively. The current method was successfully applied to screening methanolic extracts of six different J. curcas leaf genotypes (three toxic and three non-toxic). PDA and three PE deoxyphorbol metabolites were identified only from toxic genotypes, in which PDA was determined with concentration ranging from 222.19 ± 23.55 to 528.23 ± 19.72 µg/g. All these findings support that the MIPCPESI-MS method developed here has a high potential for the analysis of PEs in plant extracts enabling differentiation of toxic and non-toxic genotypes earlier in the leaves.
Assuntos
Jatropha/química , Impressão Molecular/métodos , Ésteres de Forbol/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Limite de Detecção , Modelos Lineares , Compostos Fitoquímicos/química , Polímeros , Reprodutibilidade dos TestesRESUMO
The analyses of drugs and metabolites in complex matrices have been widely studied in recent years. However, due to high levels endogenous compounds and matrix complexity, these analyses require a sample pre-treatment step. To this aim, two lab-made extractive phases were integrated to probe electrospray ionization mass spectrometry (PESI-MS) technique for direct analysis of illicit drugs in biological fluids and phorbol esters in Jatropha curcas extract. The polypyrrole (PPy) phase was electropolymerized onto a platinum wire surface by cyclic voltammetry. The molecularly imprinted polymer (MIP) was synthesized and adhered onto a stainless-steel needle with epoxy resin. The PPy-PESI-MS method showed to be linear in a concentration range from 1 to 500⯵g L-1, with accuracy values between -2.1 and 14%, and precision values between 0.8 and 10.8%. The MIP-PESI-MS method showed to be linear in a concentration range from 0.9 to 30â¯mgâ¯L-1, with accuracy values between -1.6 and -15.3%, and precision values between 4.1 and 13.5%.
Assuntos
Impressão Molecular/métodos , Preparações Farmacêuticas/análise , Preparações Farmacêuticas/isolamento & purificação , Polímeros/química , Pirróis/química , Microextração em Fase Sólida/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Cocaína/análise , Cocaína/isolamento & purificação , Voluntários Saudáveis , Humanos , Jatropha/química , Dietilamida do Ácido Lisérgico/análise , Dietilamida do Ácido Lisérgico/isolamento & purificação , Metanfetamina/análise , Metanfetamina/isolamento & purificação , N-Metil-3,4-Metilenodioxianfetamina/análise , N-Metil-3,4-Metilenodioxianfetamina/isolamento & purificação , Ésteres de Forbol/análise , Ésteres de Forbol/isolamento & purificação , Extratos Vegetais/análise , Extratos Vegetais/isolamento & purificação , Saliva/metabolismo , Aço Inoxidável/química , UrináliseRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Jatropha curcas L. is a plant with high cultural significance for quilombola communities of Oriximiná (Pará State, Brazil). Although the plant is highly toxic, its seeds are used in these communities to treat tuberculosis and related diseases and symptoms. AIM OF THE STUDY: This study was designed to provide a scientific rationale for the traditional detoxification method and use of J. curcas seeds in quilombola communities of Oriximiná. MATERIALS AND METHODS: J. curcas seeds were manually separated into testa, tegmen, endosperm, and embryo, and then methanolic extracts of each sample were prepared. The traditional preparation of J. curcas seeds consists of a water extract of endosperms that is known as "milk of pinhão-branco". The content of phorbol esters (PEs) in the extracts was analyzed by High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD). The cytotoxic activity was evaluated in human monocytic cell line THP-1 by Resazurin Reduction Assay, and antimycobacterial activity was assessed by determining Minimal Inhibitory Concentration (MIC) values against H37Rv and BCG strains using the Resazurin Microtiter Assay (REMA). RESULTS: The content analysis revealed that the distribution of PEs within the seeds is not homogeneous. High contents were found in tegmens (4.22⯱â¯0.25-15.52⯱â¯0.06â¯mg/g) and endosperms (1.61⯱â¯0.07-5.00⯱â¯0.42â¯mg/g), while concentrations found in testas and embryos were all below 0.5â¯mg/g. The traditional preparation derived from the endosperm of J. curcas contained significantly less PEs than the endosperms (0.01⯱â¯0.005â¯mg/g). Against THP-1â¯cells, all the parts of the seed showed cytotoxic activity, while the traditional preparation was considered non-cytotoxic. Nevertheless, only the tegmen and endosperm of J. curcas were considered active against M. tuberculosis and M. bovis (MICâ¯=â¯200⯵g/mL). CONCLUSION: The results of this study indicated that the traditional processing performed by the quilombola people from Oriximiná is effective in reducing the toxicity of J. curcas seeds. Although inactive against mycobacteria, the extensive use of the traditional preparation and its low toxicity encourage further studies to investigate other biological activities.
Assuntos
Jatropha , Medicina Tradicional , Ésteres de Forbol , Extratos Vegetais , Sementes/química , Antibacterianos/análise , Antibacterianos/farmacologia , Antibacterianos/toxicidade , Brasil , Sobrevivência Celular/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Mycobacterium bovis/efeitos dos fármacos , Mycobacterium bovis/crescimento & desenvolvimento , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/crescimento & desenvolvimento , Ésteres de Forbol/análise , Ésteres de Forbol/farmacologia , Ésteres de Forbol/toxicidade , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Extratos Vegetais/toxicidade , Células THP-1RESUMO
Abstract The Tehuacán-Cuicatlán Biosphere Reserve in México holds genetic richness and endemism for the genus Jatropha, but the specific use of the plants and the seed chemical profiles are still unknown. We recorded the traditional forms of use of Jatropha species, and analyzed the chemical composition of Jatropha spp. seeds. For this, a semi-structured survey was conducted in 15 towns, and 20 interviews were applied in each one between May and August 2012 to estimate the Significant Use Level and Relative Use Value per species. Besides, seeds from J. neopauciflora, J. rufescens, and J. rzedowskii were collected between August and October 2013, and to seed flour samples we determined total lipids by the Soxhlet method, crude protein by the Kjendahl method, and ashes according to AOAC methods; fatty acid profile and phorbol esters were determined by gas chromatography and by HPLC, respectively. The species J. neopauciflora, J. oaxacana, J. rufescens, J. ciliata, and J. rzedowskii, are used as traditional medicine, food, and ornaments, except for J. ciliata, for which none use was recorded. The Significant Use Level as food was found not significant, but for medicinal purposes, J. neopauciflora obtained the highest Relative Use Value (9.0 %). The latex is used to treat 13 disorders, including dental problems and oral diseases with a Significant Use Level of 32.9 %; it is also used as a hemostatic. The protein content among species varied from 23.37 to 26.06 %, and total lipids from 34.79 to 36.60 %. The principal unsaturated fatty acids were oleic (25.08 to 30.09 %) and linoleic (44.55 to 48.46 %), and the saturated fatty acids were palmitic (10.11 to 16.50 %) and stearic (9.47 to 11.15 %). Phorbol esters, the main cause of seed's toxicity, were absent in J. neopauciflora, J. rufescens, and J. rzedowskii. In conclusion, the Significant Use Level of Jatropha species studied was low, with little cultural acceptance and sporadic utilization. The Relative Use Value was important for medicinal purposes, especially for J. neopauciflora. The dehulled seeds of J. neopauciflora, J. rufescens, and J. rzedowskii are potentially useful as food, having high protein contents, and unsaturated oleic and linoleic fatty acids. With this study we report three non-toxic Jatropha species and recommend to scientifically validate the antimycotic use of J. neopauciflora latex. Rev. Biol. Trop. 66(1): 266-279. Epub 2018 March 01.
Resumen La Reserva de la Biosfera Tehuacán-Cuicatlán, México contiene riqueza genética y endemismo para el género Jatropha, pero se desconoce el uso específico de las plantas y la composición química de la semilla. Documentamos las formas tradicionales de uso de las especies de Jatropha y, analizamos la composición química de las semillas de las especies de Jatropha. En 15 comunidades se aplicaron 20 entrevistas semiestructuradas entre mayo y agosto de 2012 para estimar el Nivel de Uso Significativo y el Valor de Uso Relativo por especie. Se colectó semilla solo de J. neopauciflora, J. rufescens, y J. rzedowskii entre agosto y octubre de 2013 para determinar mediante los métodos de la AOAC, el contenido total de lípidos en una muestra de 3 g de harina mediante el método de Soxhlet, proteína cruda en una muestra de harina de 0.4 g por el método Kjendahl y ceniza en una muestra de 5 g de harina. El perfil de ácidos grasos se determinó por cromatografía de gases y el contenido de ésteres de forbol mediante HPLC. Las especies registradas fueron J. neopauciflora, J. oaxacana, J. rufescens, J. rzedowskii y J. ciliata, que se usan en medicina tradicional, alimento y como ornamental, excepto J. ciliata, para la cual no se reportó ningún uso. El Nivel de Uso Significativo como alimento no fue importante, pero para propósitos medicinales J. neopauciflora tuvo el mayor Valor de Uso Relativo (9.0 %). El látex es usado para tratar 13 trastornos, incluyendo problemas dentales y enfermedades bucales con un Nivel Significativo de Uso de 32.9 %; y también es usada como hemostático. El contenido de proteína entre especies varió de 23.37 a 26.06 % y los lípidos totales variaron de 34.79 a 36.60 %. Los principales ácidos grasos insaturados fueron oleico (25.08 a 30.09 %) y linoleico (44.55 a 48.46 %); los ácidos grasos saturados fueron palmítico (10.11 a 16.50 %) y esteárico (9.47 a 11.15 %). No se detectó la presencia de ésteres de forbol (la causa principal de la toxicidad de la semilla) en J. neopauciflora, J. rufescens, y J. rzedowskii. En conclusión, el Nivel de Uso Significativo de las especies de Jatropha fue bajo, con poca aceptación cultural y utilización esporádica. El Valor de Uso Relativo fue importante para propósitos medicinales, especialmente para J. neopauciflora. Las semillas sin testa de J. neopauciflora, J. rufescens, y J. rzedowskii son potencialmente útiles como alimento por su alto contenido de proteínas, ácidos grasos insaturados oleico y linoleico. Se identificaron tres especies no toxicas de Jatropha. Se recomienda validar científicamente el uso antimicótico del látex de J. neopauciflora.
RESUMO
BACKGROUND: Jatropha curcas L. (Jatropha) is believed to have originated from Mexico and Central America. So far, characterization efforts have focused on Asia, Africa and Mexico. Non-toxic, low phorbol ester (PE) varieties have been found only in Mexico. Differences in PE content in seeds and its structural components, crude oil and cake from Jatropha provenances cultivated in Central and South America were evaluated. Seeds were dehulled, and kernels were separated into tegmen, cotyledons and embryo for PE quantitation by RP-HPLC. Crude oil and cake PE content was also measured. RESULTS: No phenotypic departures in seed size and structure were observed among Jatropha cultivated in Central and South America compared to provenances from Mexico, Asia and Africa. Cotyledons comprised 96.2-97.5 %, tegmen 1.6-2.4 % and embryo represented 0.9-1.4 % of dehulled kernel. Total PE content of all nine provenances categorized them as toxic. Significant differences in kernel PE content were observed among provenances from Mexico, Central and South America (P < 0.01), being Mexican the highest (7.6 mg/g) and Cabo Verde the lowest (2.57 mg/g). All accessions had >95 % of PEs concentrated in cotyledons, 0.5-3 % in the tegmen and 0.5-1 % in the embryo. Over 60 % of total PE in dehulled kernels accumulated in the crude oil, while 35-40 % remained in the cake after extraction. CONCLUSIONS: Low phenotypic variability in seed physical, structural traits and PE content was observed among provenances from Latin America. Very high-PE provenances with potential as biopesticide were found in Central America. No PE-free, edible Jatropha was found among provenances currently cultivated in Central America and Brazil that could be used for human consumption and feedstock. Furthermore, dehulled kernel structural parts as well as its crude oil and cake contained toxic PE levels.
RESUMO
The latex of Euphorbia papillosa A.St.-Hil. and Sapium glandulosum (L.) Morong was phytochemically investigated by liquid chromatography/mass spectrometry. The results reveal the presence of tigliane-type diterpenoid compounds, particularly 12-deoxyphorbol esters, in both species. In addition, ingenane-type diterpene esters were found in the latex of E. papillosa. The latex of S. glandulosum showed only tigliane-type diterpenes, confirming the literature data on this genus. According to mass data, we are proposing the presence of 10 compounds derived from 12-deoxyphorbol monoesters and diesters, from one 12-deoxyphorbaldehyde, from two ingenol derivatives and from one nitrogen-containing phorbol ester in the latex of the analysed species. Considering that 12-deoxy tigliane diterpenes are described as antitumour and antiviral agents, these results indicate a pharmacological potential for these two Euphorbiaceae species.
Assuntos
Cromatografia Líquida/métodos , Diterpenos/análise , Euphorbiaceae/química , Látex/análise , Espectrometria de Massas por Ionização por Electrospray/métodosRESUMO
The crude latex of "Crown-of-Thorns" (Euphorbia milii var hislopii, syn E.splendens) is a potent plant molluscicide. For this reason, toxicological studies have been performed to evaluate the health risks posed by its use in schistosomiasis control programs. The present study is part of a more comprehensive immunotoxicological evaluation of this molluscicide. Here, we investigated the effects of E. milii latex on the proliferation of human lymphocytes in vitro. Lyophilized latex of E. milii (0, 0.5, 5, 25 and 50 µg/ml) was incubated with whole blood in the presence of proliferation stimulators, i.e. lectins (phytohemagglutinin, concanavalin A and pokeweed mitogen), as well as with human monoclonal antibody against CD3 and tetanus toxoid. Cell proliferation was measured by ³H-thymidine incorporation, and the effects of latex on mitogen-induced cell proliferation were compared to the effects of 10 ng/ml of 12-O-tetradecanoylphorbol-13-acetate (TPA). Results showed that mitogen-induced cell proliferation was markedly enhanced by E. milii latex. This synergistic effect of latex on mitogen-induced lymphocyte proliferation may be due to the presence of TPA-like phorbol esters and/or to mitogenic plant lectins.
O látexbrutoda "Coroa de Cristo" (Euphorbia miliivarhislopii, syn E.splendens) é um potente moluscicidavegetal. Neste sentido, são necessários estudos toxicológicosque visemavaliar possíveis riscos à saúdeassociados ao uso em larga escala desta espécie em áreas endêmicas para esquistossomose. O presente estudo é parte deuma avaliação mais abrangentesobre o potencial tóxico destemoluscicida. Foram investigados in vitro osefeitos dolátex da E.miliisobre a proliferação delinfócitoshumanos. O látexliofilizado (0; 0,5;5;25 e 50 µg/ml)foi incubado comsangue totalna presençade agentes mitogênicos, tais como lectinas(fitohemaglutinina, concanavalina Ae pokeweed), anticorpomonoclonalhumano anti-CD3etoxóide tetânico. A proliferação celularfoi quantificada atravésincorporaçãode ³H-timidina eos efeitos do látexnaproliferação celular induzida por agentes mitogênicosforam comparados comos efeitos de10 ng/mlde12-O-tetradecanoilforbol-13-acetato (TPA). Os resultados demonstram quea proliferação celular induzida poragentes mitogênicosfoimarcadamenteaumentada na presença do látex daE.milii.Oefeito sinérgico observado pode ser devidoà presença deésteres de forbol, como o TPA, e/oude lectinas com ação mitogênica presentes nesta espécie vegetal.
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Humanos , Masculino , Feminino , Euphorbia/metabolismo , Látex/análise , Plantas Medicinais/classificação , Ésteres de Forbol/classificação , Linfócitos/metabolismoRESUMO
Introducción: Partes aéreas de la planta Caesalpinia pulcherrima L. (Swartz) han sido usadas en medicina tradicional al sur del Departamento de Cundinamarca para el tratamiento de afecciones inflamatorias. Objetivo: Evaluar la actividad antiinflamatoria de flores, hojas y frutos verdes de Caesalpinia pulcherrima para cuantificar su actividad antiinflamatoria en modelos murinos de inflamación aguda y subcrónica. Metodología: Este estudio cuantificó la actividad antiinflamatoria de diferentes extractos de tejidos aéreos de esta especie encontrada en Colombia por dos modelos de inflamación aguda, el edema auricular inducido por TPA (acetato de 12-O-tetradecanoil-forbol) y el edema plantar inducido por carragenina; así como el modelo de inflamación sub-crónico de granuloma inducido por pellet de algodón. Resultados: Los extractos de flores mostraron la mayor actividad antiinflamatoria en el modelo del TPA, en tanto que las hojas fueron más efectivas en disminuir el granuloma, en el modelo del pellet de algodón. No se observó ninguna actividad antiinflamatoria de ningún extracto en el modelo de carragenina. Los frutos verdes no mostraron actividad en ningún modelo. Conclusión: Los resultados demostraron la efectividad que el uso etnobotánico le atribuye a esta planta. Los extractos activos obtenidos mostraron el potencial uso de esta planta en la fabricación fitoterapéuticos efectivos. Salud UIS 2011; 43 (3): 281-287.
Introduction: Aerial parts of Caesalpinia pulcherrima L. (Swartz) have been used in traditional medicine in southern Cundinamarca Department for the treatment of inflammatory diseases. Objective: Evaluate the antiinflammatory activity of flowers, leaves and green fruits of the plant to quantify inflammatory activity of acute and subchronic murine inflammation models. Methodology: This study quantified the anti-inflammatory activity of different extracts of aerial tissues of this species for two models of acute inflammation, the ear edema induced by TPA (12-O-tetradecanoyl-phorbol-13-acetate) and plantar edema induced by carrageenan, as well as the sub-chronic inflammation model of granuloma induced by cotton pellet. Results: The flower extracts proved to have the largest anti-inflammatory activity in the TPA model, while the leaves were more effective in reducing the granuloma in the cotton pellet model. There was no anti-inflammatory activity of any extract carrageenan model. The green fruits showed no activity in any model. Conclusion: The results demonstrated the effectiveness of the ethnobotanical use attributed to this plant. The active extracts obtained showed the potential use of this plant in making safe and effective phytomedicines. Salud UIS 2011; 43 (3): 281-287.