RESUMEN
Polyphenols from residual non-food grade Salicornia ramosissima have health-promoting effects in feed, food, or nutraceutical applications. Therefore, the isolation of polyphenols is of interest from a series of environmentally friendly isolation methods with recyclable solvents. The isolation of polyphenols from non-food grade S. ramosissima was investigated using sequential membrane filtration with and without acid pretreatment, liquid-liquid extraction, resin adsorption, and centrifugal partition chromatography (CPC); analyzed by the Folin-Ciocalteu assay for total polyphenols; and finally analyzed using UPLC-TQMS in negative ion-spray mode for detection of 14 polyphenols. Sequential membrane filtration and acid hydrolysis indicated the polyphenols forming complexes with other compounds, retaining the polyphenols in the retentate fraction of large molecular weight cut-off membrane sizes. Conventional liquid-liquid extraction using sequential ethyl acetate and n-butanol showed most polyphenols were extracted, apart from chlorogenic acids, indicating a low isolation efficiency of higher polarity polyphenols. Analysis of the extract after resin adsorption by Amberlite XAD-4 resin showed high efficiency for separation, with 100% of polyphenols adsorbed to the resin after 13 bed volumes and 96.7% eluted from the resin using ethanol. CPC fractionations were performed to fractionate the concentrated extract after resin adsorption. CPC fractionations of the 14 polyphenols were performed using an organic or aqueous phase as a mobile phase. Depending on the mobile phase, different compounds were isolated in a high concentration. Using these easily scalable methods, it was possible to comprehensively study the polyphenols of interest from S. ramosissima and their isolation mechanics. This study will potentially lead the way for the large-scale isolation of polyphenols from S. ramosissima and other complex halophytes. The compounds of the highest concentration after CPC fractionation were isoquercitrin and hyperoside (155.27 mg/g), chlorogenic acid (85.54 mg/g), cryptochlorogenic acid (101.50 mg/g), and protocatechuic acid (398.67 mg/g), and further isolation using CPC could potentially yield novel polyphenol nutraceuticals.
Asunto(s)
Chenopodiaceae , Polifenoles , Plantas Tolerantes a la Sal , Ácido Clorogénico , SolventesRESUMEN
PURPOSE: We assessed the accuracy of lens power calculation in highly myopic patients implanting plus and minus intraocular lenses (IOL). METHODS: We included 58 consecutive, myopic eyes with an axial length (AL) > 26.0 mm, undergoing phacoemulsification and IOL implantation following biometry using the IOLMaster 500. For lens power calculation, the Haigis formula was used in all cases. For comparison, refraction was back-calculated using the Barrett Universal II (Barrett), Holladay I, Hill-RBF (RBF) and SRK/T formulae. RESULTS: The mean axial length was 30.17 ± 2.67 mm. Barrett (80%), Haigis (87%) and RBF (82%) showed comparable numbers of IOLs within 1 diopter (D) of target refraction. Visual acuity (BSCVA) improved (p < 0.001) from 0.60 ± 0.35 to 0.29 ± 0.29 logMAR (> 28-days postsurgery). The median absolute error (MedAE) of Barrett 0.49 D, Haigis 0.38, RBF 0.44 and SRK/T 0.44 did not differ. The MedAE of Haigis was significantly smaller than Holladay (0.75 D; p = 0.01). All median postoperative refractive errors (MedRE) differed significantly with the exception of Haigis to SRK/T (p = 0.6): Barrett - 0.33 D, Haigis 0.25, Holladay 0.63, RBF 0.04 and SRK/T 0.13. Barrett, Haigis, Holladay and RBF showed a tendency for higher MedAEs in their minus compared to plus IOLs, which only reached significance for SRK/T (p = 0.001). Barrett (p < 0.001) and RBF (p = 0.04) showed myopic, SRK/T (p = 002) a hyperopic shift in their minus IOLs. CONCLUSIONS: In highly myopic patients, the accuracies of Barrett, Haigis and RBF were comparable with a tendency for higher MedAEs in minus IOLs. Barrett and RBF showed myopic, SRK/T a hyperopic shift in their minus IOLs.
Asunto(s)
Lentes Intraoculares , Miopía , Facoemulsificación , Biometría , Humanos , Interferometría , Implantación de Lentes Intraoculares , Miopía/diagnóstico , Óptica y Fotónica , Refracción Ocular , Estudios RetrospectivosRESUMEN
The ethyl acetate and n-butanolic subfractions of Agelanthus dodoneifolius were investigated for their antioxidant and antimyeloperoxidase (MPO) activities. The reactive oxygen species (ROS) generation was assessed by lucigenin-enhanced chemiluminescence (CL) and dichlorofluorescein- (DCF-) induced fluorescence techniques from phorbol myristate acetate- (PMA-) stimulated equine neutrophils and human myeloid cell line HL-60, respectively. In parallel, the effects of the tested subfractions were evaluated on the total MPO release by stimulated neutrophils and on the specific MPO activity by means of immunological assays. The results showed the potent activity of the butanolic subfraction, at least in respect of the chemiluminescence test (IC50 = 0.3 ± 0.1 µg/mL) and the ELISA and SIEFED assays (IC50 = 2.8 ± 1.2 µg/mL and 1.3 ± 1.0 µg/mL), respectively. However, the ethyl acetate subfraction was found to be the most potent in the DCF assay as at the highest concentration, DCF fluorescence intensity decreases of about 50%. Moreover, we demonstrated that the ethyl acetate subfraction was rich in catechin (16.51%) while it was not easy to identify the main compounds in the butanolic subfraction using the UPLC-MS/MS technique. Nevertheless, taken together, our results provide evidence that Agelanthus dodoneifolius subfractions may represent potential sources of natural antioxidants and of antimyeloperoxidase compounds.
RESUMEN
Agelanthus dodoneifolius DC Danser (Loranthaceae) is used for the treatment of various diseases including asthma. The aqueous and hydroalcoholic extracts have been reported to have anti-inflammatory, spasmolytic and bronchorelaxant activities. The present study investigates the effects of the aqueous decoction and the diethyl ether, ethyl acetate and butanolic fractions of Agelanthus dodoneifolius DC Danser (Loranthaceae) on reactive oxygen species (ROS) production and myeloperoxidase (MPO) release by phorbol 12-myristate 13-acetate (PMA)-stimulated equine neutrophils and on purified equine MPO activity. ROS production and MPO release by the PMA-stimulated neutrophils were measured by the lucigenin-enhanced chemiluminescence and ELISA assays, respectively. Specific immunological extraction followed by enzymatic detection (SIEFED) was used to specifically measure the equine MPO activity. Identification and quantification of the individual and total phenolic and flavonoid compounds were performed using UPLC-MS/MS equipment and colorimetric methods involving Folin-Ciocalteu and AlCl3, respectively. All the tested extracts displayed dose-dependent inhibitory effects on the oxidant activities of neutrophils; a stronger effect was observed with the organic fractions than the aqueous decoction. These findings could be correlated with a high content of phenolic and flavonoid compounds. The results confirm the previously shown anti-inflammatory effect of Agelanthus dodoneifolius and its potential use for the treatment of neutrophil-dependent inflammatory diseases.
Asunto(s)
Loranthaceae/química , Neutrófilos/efectos de los fármacos , Peroxidasa/metabolismo , Extractos Vegetales/farmacología , Animales , Antiinflamatorios/farmacología , Supervivencia Celular/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Flavonoides/química , Flavonoides/farmacología , Caballos , Activación Neutrófila/efectos de los fármacos , Neutrófilos/enzimología , Fenoles/química , Fenoles/farmacología , Extractos Vegetales/química , Hojas de la Planta/química , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Resveratrol is a polyphenolic antioxidant present in beverage and food known for its multiple protective effects. We report the inhibitory effects of resveratrol on equine myeloperoxidase (MPO), a hemic peroxidase present in the granules of the neutrophils involved in the inflammatory response. Resveratrol inhibited the production of reactive oxygen species (ROS) by stimulated equine neutrophils by acting as a direct scavenger of the ROS released by the cells but did not modify the degranulation of the stimulated neutrophils as the amounts of released MPO were unchanged. Resveratrol strongly inhibited the chlorination, oxidation, and nitration activities of MPO in a dose-dependent manner. By an original technique of specific immunological extraction followed by enzymatic detection (SIEFED), we demonstrated that resveratrol inhibited the peroxidasic activity of the MPO measured by a direct interaction such as the fixation of resveratrol on the enzyme. The observation of a decrease of the accumulation of compound II suggested that resveratrol acts as an electron donor for MPO reduction.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Caballos , Neutrófilos/enzimología , Peroxidasa/antagonistas & inhibidores , Estilbenos/farmacología , Animales , Peroxidasa/metabolismo , Especies Reactivas de Oxígeno/metabolismo , ResveratrolRESUMEN
We report here on the role of copper (II) salts on the acceleration of peroxynitrite (ONOO-) decomposition and ONOO- reaction with the anaesthetic agent propofol (2,6-diisopropylphenol) in alkaline medium. We observed a strong acceleration of the ONOO- decomposition in alkaline medium in the presence of copper (I and II) salts. After 18 h of ONOO- reaction with propofol, we observed nitrosated, nitrated, and oxidized (quinone and diphenylquinone) derivatives of propofol, but in the presence of Cu(II) (20% molar vs ONOO-), the yields of quinone and nitrosopropofol strongly increased. We also observed that the temperature and the atmosphere influenced the effects of Cu(II) on ONOO- reactions with propofol: low temperatures promoted nitrosation and high temperatures promoted oxidation; O2 atmosphere increased the general reactivity and the yield of nitrated and oxidized products. We highlighted the influence of Cu(II) salts on the radical character of the reaction by direct EPR technique. The exact mechanism of the Cu(II) catalysis remains unexplained, but we suggest the formation of a copper complex with propofol or, more probably, the oxidation of ONOO- into ONOO. by copper ions promoting the formation of quinone and nitrosopropofol according to a previously reported mechanism [M. Cudic, C. Ducrocq, Transformations of 2,6-diisopropylphenol by NO-derived nitrogen oxides, particularly peroxynitrite, Nitric Oxide 4 (2000) 147-156].
Asunto(s)
Álcalis/química , Cobre/química , Ácido Peroxinitroso/química , Propofol/química , Sales (Química)/química , Bicarbonatos/química , Dióxido de Carbono/química , Catálisis , Espectroscopía de Resonancia por Spin del Electrón , Concentración de Iones de Hidrógeno , Estructura Molecular , Nitrógeno/química , Dióxido de Nitrógeno/química , Oxígeno/química , Soluciones , TemperaturaRESUMEN
Chronic inflammation through foam cells and macrophages is important in atherosclerosis development, and can be considered as therapeutic targets. Cyclooxygenase and NADPH-oxidase were expressed within atherosclerotic lesions. Reactive oxygen species produced by NADPH oxidase were found to trigger the cyclooxygenase-2 expression. The effects of preferential COX-2 inhibitors on ROS produced by Chlamydia-primed human monocytes (THP-1 cells) were evaluated by fluorescence, chemiluminescence, oxymetry, and EPR spin trapping. Fluorescence assays showed an increased production of ROS with Chlamydia versus cells primed by 10(-8)M PMA. COX-2 inhibitors inhibited in a dose-dependent manner the luminol-enhanced CL while ibuprofen and diclofenac increased the chemiluminescence response. By EPR spin trapping, COX-2 inhibitors, ibuprofen, and diclofenac, exhibited a dose-dependent inhibiting effect (10 and 100muM) on the EPR signal appearance. Our cell model combining EPR, chemiluminescence, and oxymetry appeared relevant to study the modulating effects of preferential COX-2 inhibitors on the cell oxidant activity and chronic inflammatory diseases.
Asunto(s)
Chlamydophila pneumoniae/patogenicidad , Inhibidores de la Ciclooxigenasa/farmacología , Monocitos/metabolismo , Monocitos/microbiología , Especies Reactivas de Oxígeno/metabolismo , Arteriosclerosis/metabolismo , Arteriosclerosis/prevención & control , Diferenciación Celular , Línea Celular , Relación Dosis-Respuesta a Droga , Radicales Libres/metabolismo , Humanos , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Macrófagos/microbiología , Monocitos/citología , Monocitos/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Four analogues of Ebselen were synthesized and their glutathione peroxidase activity and antioxidant property evaluated and compared to Ebselen. Among the studied compounds, only diselenide [3] exhibited both glutathione peroxidase activity and radical-scavenging capability. Compounds [3] and [4] showed a strong inhibitory effect (53% and 43%, respectively) on the lipid peroxidation of linoleic acid compared to Ebselen and selenide derivatives ([1] and [2]) which were less active (28%, 26% and 18% inhibition, respectively). A concentration-dependent inhibitory effect was also found in the model of the formation of ABTS*+ radical cation: 65% and 89% inhibition for compound [3] at 10(-4) M and 5 x 10(-5) M, respectively, and 68% and 90% for compound [4], compared to 14% and 52% inhibition for Ebselen and the diselenides [1] and [2] (29%, 46% and 45%, 68%, respectively). By EPR spin trapping technique, the following inhibitory profile of the Ebselen analogues was observed towards the formation of thiyl radicals: Ebselen = [3]>[1]>[2]>[4]. Studies with compound [3] are in progress on oxidative stress cell models.
Asunto(s)
Antioxidantes/metabolismo , Azoles/metabolismo , Glutatión Peroxidasa/metabolismo , Compuestos de Organoselenio/metabolismo , Espectroscopía de Resonancia por Spin del Electrón , Técnicas In Vitro , Isoindoles , Peroxidación de LípidoRESUMEN
The reaction between the anaesthetic agent 2,6-diisopropylphenol (propofol, PPF) and singlet oxygen (1O2) has been investigated in aqueous solution by means of HPLC, GC, absorption spectroscopy and laser flash photolysis with infrared luminescence detection. The rate constants for the physical and chemical quenching of 1O2 by PPF (kPPF) are found to be 2.66 x 10(5) M(-1) s(-1) and approximately 3.2 x 10(6) M(-1) s(-1) in CD3OD and D2O-CD3OD (75:25 v/v), respectively. The reaction of propofol with singlet oxygen produced by light irradiation of Rose Bengal leads essentially to two reaction products, 2,6-diisopropyl-p-benzoquinone and 3,5,3',5'-tetraisopropyl-(4,4')-diphenoquinone that are unambiguously identified from comparison with authentic samples.
Asunto(s)
Anestésicos Intravenosos/química , Propofol/química , Oxígeno Singlete/química , Cromatografía de Gases , Cromatografía Líquida de Alta Presión , CinéticaRESUMEN
We reported here the reaction, in acidic conditions, of peroxynitrite (ONOO(-)) with the anaesthetic agent propofol (2,6-diisopropylphenol, PPF). The most interesting finding is that peroxynitrite is able to nitrate and to oxidize propofol leading to 4-nitropropofol, quinone, and diphenylquinone as the major products. More surprisingly, we also found that peroxynitrite is capable of halogenating propofol in the presence of halide ions, suggesting the formation of nitrosyl chloride (NOCl) or nitryl chloride (NO(2)Cl) from the reaction of peroxynitrite with chloride ions. A significant enhancement of the halogenation yield is observed with a simultaneous decrease of the yields of the other products in the presence of methanol or hydrogen peroxide. Increased halogenation of PPF probably results from the formation of peroxynitrate (O(2)NOO(-)), that further oxidizes chloride ions in hypochlorous acid (HOCl) or molecular chlorine (Cl(2)). Spontaneous decay of peroxynitrate is relatively slow in acidic medium, thus explaining the decrease of the yields of the other products. By direct EPR techniques, we also observed that this reaction occurs via a radical pathway.
Asunto(s)
Anestésicos Intravenosos/química , Ácido Peroxinitroso/química , Propofol/química , Sistema Nervioso Central/química , Sistema Nervioso Central/metabolismo , Radicales Libres/química , Halógenos/química , Humanos , Concentración de Iones de Hidrógeno , Óxidos de Nitrógeno/química , Oxidación-ReducciónRESUMEN
PURPOSE: To compare the surgically induced corneal astigmatism after unsutured temporal and nasal unsutured limbal tunnel incisions. SETTING: Departments of Ophthalmology, Marienhospital, Aachen, and Johann Wolfgang Goethe-University, Frankfurt am Main, Germany. METHODS: In a prospective clinical study, 42 eyes of 21 patients with a mean age of 75.1 years had phacoemulsification and implantation of a foldable hydrophobic acrylic intraocular lens using a 3.6 to 3.8 mm unsutured limbal tunnel incision. The right eye always received a temporal incision and the left eye, a nasal incision. Computerized videokeratography was performed preoperatively and 2 weeks and 6 months postoperatively. Surgically induced astigmatism was calculated by vector analysis using the Holladay-Cravy-Koch formula. The 2 groups were compared using a paired Wilcoxon test. RESULTS: The mean surgically induced corneal astigmatism in the temporal incision group was 0.62 diopters (D) +/- 0.48 (SD) 2 weeks postoperatively and 0.47 +/- 0.32 D at 6 months and in the nasal incision group, 1.55 +/- 0.84 D and 1.05 +/- 0.57 D, respectively. The difference between the groups was statistically significant (P <.05). CONCLUSIONS: There was a highly statistically significant difference in surgically induced corneal astigmatism after temporal and nasal unsutured limbal tunnel incisions. The degree of induced astigmatism and the difference between the temporal and the nasal incisions decreased over time. A nasal tunnel incision is not appropriate for astigmatism-neutral surgery.