RESUMEN
Dietary rumen-protected polyunsaturated fatty acids (PUFAs) rich in linoleic acid (LA) may affect embryo yield, and LA can modulate the molecular mechanisms of lipid uptake in bovine blastocysts produced in vitro. In embryos, membrane lipids, such as phosphatidylcholines (PCs) and sphingomyelins (SMs), affect cryopreservation success. The aim of the present study was to evaluate embryonic developmental rates after the IVF of oocytes retrieved from Nellore heifers fed for approximately 90 days with rumen-protected PUFAs rich in LA. In addition, we evaluated embryo cryotolerance and the membrane structure lipid composition using matrix-assisted laser desorption ionisation mass spectrometry of fresh and vitrified embryos. Embryo development to the blastocyst stage (mean 43.2%) and embryo survival after vitrification and warming (mean 79.3%) were unaffected by diet. The relative abundance of one lipid species (PC ether (PCe; 38:2, which means that this lipid has 38 carbon atoms and 2 double bonds in the fatty acyl residues) was increased after PUFAs supplementation. However, 10 ions were affected by cryopreservation; ions consistent with PC 32:0, PC 34:1, SM 24:1, PC 40:6 or PC 42:9, PC plasmalogen (PCp) 44:10 or PC 42:7, triacylglycerol (TAG) 54:9 and a not assigned ion (m/z 833.2) were lower in blastocysts that survived to the cryopreservation process compared with fresh blastocysts, whereas the abundance of the ions PC 36:3 or PC 34:0, PCe 38:2 or PC 36:6 and PC 36:5 or PCe 38:1 were increased after cryopreservation. Thus, the results demonstrate that the mass spectrometry profiles of PC, SM and TAG species differ significantly in bovine blastocysts upon cryopreservation. Because the lipid ion abundances of fresh and vitrified-warmed embryos were distinct, they can be used as potential markers of post-cryopreservation embryonic survival.
Asunto(s)
Criopreservación/veterinaria , Grasas Insaturadas en la Dieta/administración & dosificación , Ectogénesis , Embrión de Mamíferos/metabolismo , Fenómenos Fisiologicos Nutricionales Maternos , Lípidos de la Membrana/metabolismo , Oocistos/metabolismo , Animales , Animales Endogámicos , Blastocisto , Brasil , Bovinos , Estudios Cruzados , Embrión de Mamíferos/citología , Femenino , Fertilización In Vitro/veterinaria , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Ácido Linoleico/administración & dosificación , Ácido Linoleico/metabolismo , Masculino , Lípidos de la Membrana/química , Oocistos/citología , Oocistos/aislamiento & purificación , Recuperación del Oocito/veterinaria , Plasmalógenos/química , Plasmalógenos/metabolismo , Preservación de Semen/veterinaria , VitrificaciónRESUMEN
This study aimed to evaluate the impact of vitrification on membrane lipid profile obtained by mass spectrometry (MS) of in vitro-produced bovine embryos. Matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) has been used to obtain individual embryo membrane lipid profiles. Due to conditions of analysis, mainly membrane lipids, most favorably phosphatidylcholines (PCs) and sphingomyelins (SMs) have been detected. The following ions described by their mass-to-charge ratio (m/z) and respective attribution presented increased relative abundance (1.2-20×) in the vitrified group: 703.5 [SM (16:0) + H]+; 722.5 [PC (40:3) + Na]+; 758.5 [PC (34:2) + H]+; 762.5 [PC (34:0) + H]+; 790.5 [PC (36:0) + H]+ and 810.5 [PC (38:4) + H]+ and/or [PC (36:1) + Na]+. The ion with a m/z 744.5 [PCp (34:1) and/or PCe (34:2)] was 3.4-fold more abundant in the fresh group. Interestingly, ions with m/z 722.5 or 744.5 indicate the presence of lipid species, which are more resistant to enzymatic degradation as they contain fatty acyl residues linked through ether type bonds (alkyl ether or plasmalogens, indicated by the lowercase 'e' and 'p', respectively) to the glycerol structure. The results indicate that cryopreservation impacts the membrane lipid profile, and that these alterations can be properly monitored by MALDI-MS. Membrane lipids can therefore be evaluated by MALDI-MS to monitor the effect of cryopreservation on membrane lipids, and to investigate changes in lipid profile that may reflect the metabolic response to the cryopreservation stress or changes in the environmental conditions.
Asunto(s)
Biomarcadores/análisis , Embrión de Mamíferos/metabolismo , Fertilización In Vitro/veterinaria , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Lípidos de la Membrana/análisis , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/veterinaria , Vitrificación , Animales , Bovinos , Embrión de Mamíferos/citología , FemeninoRESUMEN
Commercial lecithins are composed mainly of phospholipids and triacylglycerols. The analysis of the commercial lecithins, including their fraction of phospholipids, normally involves laborious and expensive protocols. Easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is shown to be an efficient technique for the analysis of lipids. Samples of commercial lecithins including standards, refined, deoiled and modified soy lecithin were tested. Characteristic profiles of phosphatidylcholines and triacylglycerols are detected by EASI(+)-MS, whereas EASI(-)-MS provided phosphatidylethanolamines, glycophospholipids and free fatty acids profiles. Acetylated lecithins also displayed characteristic acetylated derivatives. EASI-MS data was also compared to MALDI-MS, and found to display richer compositional information. The industrial process applied to lecithin fabrication was also characterised via typical EASI-MS profiles. EASI-MS both in its positive and negative ion modes offers a direct, fast and efficient technique able to characterise commercial lecithin.
Asunto(s)
Lecitinas/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos , Lecitinas/economía , Espectrometría de Masa por Ionización de Electrospray/instrumentaciónRESUMEN
Venturi easy ambient sonic spray ionization mass spectrometry in both its liquid (V(L) -EASI-MS) and solid sample modes (V(S) -EASI-MS) is shown to provide nearly immediate and secure typification of woods, as demonstrated for Mahogany, an endangered and most valuable type of tropical wood. This reddish wood displays unique phytochemical markers (phragmalin-type limonoids) which are rapidly detected from the wood surface by V(S) -EASI-MS or from a simple methanol extract of a tiny wood chip by V(L) -EASI-MS. Unique profiles were obtained for Mahogany (Swietenia macrophylla) whereas genuine samples of six other similar types of woods, which are commonly falsified by artificial coloring and commercialized as Mahogany, display also typical but dissimilar pythochemical profiles as compared to that of the authentic wood. Variable and atypical chemical profiles were observed for artificially colored woods. Secure chemical characterization via V(S) -EASI-MS or V(s) -EASI-MS fingerprints of Mahogany and other types of woods with similar appearance should help to control the illegal logging and trade of this and other endangered woods and their falsification, and to create certified standards.
Asunto(s)
Meliaceae/química , Espectrometría de Masa por Ionización de Electrospray/métodos , Madera/clasificación , Fraccionamiento Químico , Especies en Peligro de Extinción , Limoninas/análisis , Metanol , Árboles , Madera/análisis , Madera/químicaRESUMEN
BACKGROUND: To overcome the problem of increasing drug resistance, traditional medicines are an important source for potential new anti-malarials. Caesalpinia pluviosa, commonly named "sibipiruna", originates from Brazil and possess multiple therapeutic properties, including anti-malarial activity. METHODS: Crude extract (CE) was obtained from stem bark by purification using different solvents, resulting in seven fractions. An MTT assay was performed to evaluate cytotoxicity in MCF-7 cells. The CE and its fractions were tested in vitro against chloroquine-sensitive (3D7) and -resistant (S20) strains of Plasmodium falciparum and in vivo in Plasmodium chabaudi-infected mice. In vitro interaction with artesunate and the active C. pluviosa fractions was assessed, and mass spectrometry analyses were conducted. RESULTS: At non-toxic concentrations, the 100% ethanolic (F4) and 50% methanolic (F5) fractions possessed significant anti-malarial activity against both 3D7 and S20 strains. Drug interaction assays with artesunate showed a synergistic interaction with the F4. Four days of treatment with this fraction significantly inhibited parasitaemia in mice in a dose-dependent manner. Mass spectrometry analyses revealed the presence of an ion corresponding to m/z 303.0450, suggesting the presence of quercetin. However, a second set of analyses, with a quercetin standard, showed distinct ions of m/z 137 and 153. CONCLUSIONS: The findings show that the F4 fraction of C. pluviosa exhibits anti-malarial activity in vitro at non-toxic concentrations, which was potentiated in the presence of artesunate. Moreover, this anti-malarial activity was also sustained in vivo after treatment of infected mice. Finally, mass spectrometry analyses suggest that a new compound, most likely an isomer of quercetin, is responsible for the anti-malarial activity of the F4.
Asunto(s)
Antimaláricos/administración & dosificación , Antimaláricos/farmacología , Caesalpinia/química , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacología , Animales , Antimaláricos/aislamiento & purificación , Antimaláricos/toxicidad , Artemisininas/farmacología , Artesunato , Brasil , Línea Celular , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Sinergismo Farmacológico , Humanos , Malaria/tratamiento farmacológico , Malaria/parasitología , Ratones , Ratones Endogámicos C57BL , Corteza de la Planta/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/toxicidad , Plantas Medicinales/química , Plasmodium chabaudi/efectos de los fármacos , Plasmodium falciparum/efectos de los fármacos , Quercetina/administración & dosificación , Quercetina/aislamiento & purificación , Quercetina/farmacología , Quercetina/toxicidad , Enfermedades de los Roedores/tratamiento farmacológico , Enfermedades de los Roedores/parasitologíaRESUMEN
Although the production of patulin in apple fruits is mainly by Penicillium expansum, there is no information on the ability of heat resistant moulds that may survive pasteurization to produce this mycotoxin in juice packages during storage and distribution. In this study, the production of patulin by Byssochlamys spp (Byssochlamys nivea FRR 4421, B. nivea ATCC 24008 and Byssochlamys fulva IOC 4518) in cloudy and clarified apple juices packaged in laminated paperboard packages or in polyethylene terephthalate bottles (PET) and stored at both 21 degrees C and 30 degrees C, was investigated. The three Byssochlamys strains were able to produce patulin in both cloudy and clarified apple juices. Overall, the lower the storage temperature, the lower the patulin levels and mycelium dry weight in the apple juices (p<0.05). The greatest variations in pH and degrees Brix were observed in the juices from which the greatest mycelium dry weights were recovered. The maximum levels of patulin recovered from the juices were ca. 150 microg/kg at 21 degrees C and 220 microg/kg at 30 degrees C. HPLC-UV, HPCL-DAD and mass spectrometry analyses confirmed the ability of B. fulva IOC 4518 to produce patulin. Due to the heat resistance of B. nivea and B. fulva and their ability to produce patulin either in PET bottles or in laminated paperboard packages, the control of contamination and the incidence of these fungi should be a matter of concern for food safety. Control measures taken by juice industries must also focus on controlling the ascospores of heat resistant moulds.
Asunto(s)
Bebidas/microbiología , Byssochlamys/metabolismo , Embalaje de Alimentos , Malus/microbiología , Patulina/metabolismo , Contaminación de Alimentos/análisis , Patulina/análisis , TemperaturaRESUMEN
Phytochemical investigation of the bark of Guatteria hispida afforded three new alkaloids, 9-methoxy-O-methylmoschatoline (1), 9-methoxyisomoschatoline (2), and isocerasonine (3), along with 10 known alkaloids, 8-oxopseudopalmatine (4), O-methylmoschatoline (5), lysicamine (6), liriodenine (7), 10-methoxyliriodenine (8), nornuciferine (9), anonaine (10), xylopine (11), coreximine (12), and isocoreximine (13). The major compounds, 2, 6, 12, and 13, showed significant antioxidant capacity in the ORAC(FL) assay. Compounds 5, 6, and 7 were active against S. epidermidis and C. dubliniensis, with MIC values in the range 12.5-100 microg mL(-1).
Asunto(s)
Alcaloides/aislamiento & purificación , Alcaloides/farmacología , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Antifúngicos/aislamiento & purificación , Antifúngicos/farmacología , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Guatteria/química , Plantas Medicinales/química , Alcaloides/química , Antibacterianos/química , Antifúngicos/química , Antioxidantes/química , Aporfinas/química , Brasil , Candida/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Corteza de la Planta/química , Pseudomonas aeruginosa/efectos de los fármacos , Staphylococcus/efectos de los fármacosRESUMEN
A fast and reliable method is presented for the analysis of vegetable oils. Easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is shown to efficiently desorb and ionize the main oil constituents from an inert surface under ambient conditions and to provide comprehensive triacylglyceride (TAG) and free fatty acid (FFA) profiles detected mainly as either [TAG + Na](+) or [FFA-H](-) ions. EASI(+/-)-MS analysis is simple, easily implemented, requires just a tiny droplet of the oil and is performed without any pre-separation or chemical manipulation. It also causes no fragmentation of TAG ions hence diacylglyceride (DAG) and monoacylglyceride (MAG) profiles and contents can also be measured. The EASI(+/-)-MS profiles of TAG and FFA permit authentication and quality control and can be used, for instance, to access levels of adulteration, acidity, oxidation or hydrolysis of vegetable oils in general.
Asunto(s)
Ácidos Grasos/química , Aceites de Plantas/química , Espectrometría de Masa por Ionización de Electrospray/instrumentación , Triglicéridos/química , Espectrometría de Masa por Ionización de Electrospray/métodos , UltrasonidoRESUMEN
Two new lignans, magnovatins A (1) and B (2), along with nine known compounds, were isolated from the leaves of Magnolia ovata. The known compounds were identified as acuminatin (3), licarin A (4), kadsurenin M, 4-O-demethylkadsurenin M, oleiferin A, oleiferin C, spathulenol, parthenolide, and 11,13-dehydrocompressanolide. In addition, compounds 1 and 2 yielded four new derivatives (1a, 1b, 2a, and 2b). The structures of the new compounds were established on the basis of spectrometric data evaluation. Free-radical scavenging and antimicrobial activities of the major compunds 1, 3, and 4 were investigated.
Asunto(s)
Antibacterianos/aislamiento & purificación , Depuradores de Radicales Libres/aislamiento & purificación , Magnolia/química , Plantas Medicinales/química , Antibacterianos/química , Antibacterianos/farmacología , Brasil , Escherichia coli/efectos de los fármacos , Flavonoides/aislamiento & purificación , Flavonoides/farmacología , Depuradores de Radicales Libres/química , Depuradores de Radicales Libres/farmacología , Glicósidos/aislamiento & purificación , Glicósidos/farmacología , Lignanos/aislamiento & purificación , Lignanos/farmacología , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Hojas de la Planta/química , Pseudomonas aeruginosa/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Staphylococcus epidermidis/efectos de los fármacos , Levaduras/efectos de los fármacosRESUMEN
Amazonian oils and fats display unique triacylglycerol (TAG) profiles and, because of their economic importance as renewable raw materials and use by the cosmetic and food industries, are often subject to adulteration and forgery. Representative samples of these oils (andiroba, Brazil nut, buriti, and passion fruit) and fats (cupuaçu, murumuru, and ucuúba) were characterized without pre-separation or derivatization via dry (solvent-free) matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Characteristic profiles of TAG were obtained for each oil and fat. Dry MALDI-TOF MS provides typification and direct and detailed information, via TAG profiles, of their variable combinations of fatty acids. A database from spectra could be developed and may be used for their fast and reliable typification, application screening, and quality control.