Determination of phenols, flavones, and lignans in virgin olive oils by solid-phase extraction and high-performance liquid chromatography with diode array ultraviolet detection

A simple analytical method for the quantitative determination of phenols, flavones, and lignans in virgin olive oils was developed. The polar fraction was isolated from small amounts of oil sample (2.5 g) by solid-phase extraction (SPE) using diol-phase cartridges, and the extract was analyzed by reversed-phase HPLC coupled with diode array UV detection. Chromatographic separation of pinoresinol, cinnamic acid, and 1-acetoxypinoresinol was achieved. Repeatability (RSD < 6.5%), recovery (> 90%), and response factors for each identified component were determined. SPE on amino-phase cartridges was used for isolating acidic phenols and as an aid for phenol identification. For the first time, 2-(4-hydroxyphenyl)ethyl acetate was detected in olive oils. The aldehydic structure of the ligstroside aglycon was confirmed by NMR spectroscopy. The colorimetric determination of total o-diphenolic compounds by reaction with molybdate was consistent with their HPLC determination. Differences between results obtained by liquid-liquid extraction and SPE were not statistically significant.     

Relationships between oxidative stability, triacylglycerol composition, and antioxidant content in olive oil matrices

In olive oils, relationships between oxidative stability, glyceridic composition, and antioxidant content were investigated. Lipid matrices, obtained by purification of olive and high-oleic sunflower oils, were spiked with hydroxytyrosol, alpha-tocopherol, and mixtures of them and then subjected to oxidation in a Rancimat apparatus at 100 degrees C. At the same concentration of antioxidants, induction time (IT) decreased as the unsaturation rate of the matrix increased, but only fair correlations were found with fatty acid composition. Oxidative susceptibility (OS(TAG)) was calculated as a function of the relative oxidation rate of the triacylglycerols, and a linear relationship-IT (h) = (a + b)OS(TAG)-between induction time and this parameter showed a good correlation coefficient (r > 0.990, p < 0.001). In the case of matrices with a single antioxidant, origin ordinate (a) and slope (b) can be calculated as a function of the antioxidant concentration. In matrices spiked with mixtures of hydroxytyrosol and alpha-tocopherol, a simple relationship between the coefficients a and b and the concentration of antioxidants cannot be established because additive and subtractive effects occur depending on the relative concentrations of both antioxidants. However, approximate values for these coefficients can be obtained, allowing the estimation of the oil stability. In various olive oils, an acceptable agreement was found between the IT experimentally determined and that calculated from the oil composition. These results confirmed that the Rancimat stability of olive oils mainly depends on triacylglycerol composition and concentrations of o-diphenols and alpha-tocopherol.     

The composition of grain and forage from glyphosate tolerant wheat MON 71800 is equivalent to that of conventional wheat (Triticum aestivum L.)

Glyphosate tolerant wheat MON 71800, simply referred to as MON 71800, contains a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) protein from Agrobacterium sp. strain CP4 (CP4 EPSPS) that has a reduced affinity for glyphosate as compared to the endogenous plant EPSPS enzyme. The purpose of this work was to evaluate the compositional equivalence of MON 71800 to its nontransgenic parent as well as to conventional wheat varieties. The compositional assessment evaluated the levels of proximates, amino acids, fatty acids, minerals, vitamins, secondary metabolites, and antinutrients in wheat forage and grain grown during two field seasons across a total of eight sites in the United States and Canada. These data demonstrated that with respect to these important nutritional components, the forage and grain from MON 71800 were equivalent to those of its nontransgenic parent and commercial wheat varieties. These data, together with the previously established safety of the CP4 EPSPS protein, support the conclusion that glyphosate tolerant wheat MON 71800 is as safe and nutritious as commercial wheat varieties.     

Anomalous polarization profiles in sunspots: possible origin of umbral flashes

We present time-series spectropolarimetric observations of sunspots in the Ca II infrared triplet lines, which show a periodic occurrence of anomalous, asymmetric, circular polarization profiles in the umbral chromosphere. The profiles may be caused by the periodic development of an unresolved atmospheric component in a downward flowing magnetized environment. This active component with upward directed velocities as high as 10 kilometers per second is connected to the umbral flash (UF) phenomenon. We can explain the observations with a semiempirical model of the chromospheric oscillation and of the sunspot magnetized atmospheric plasma during a UF event.     

Effects of ultra-high-pressure homogenization treatment on the lipolysis and lipid oxidation of milk during refrigerated storage

Free fatty acid (FFA) release and quantification and lipid oxidation extent of ultra-high-pressure homogenized (UHPH) milk samples were evaluated to assess the effect of UHPH on the susceptibility of milk lipids to lipolysis and oxidation. Milk was UHPH-treated at 200 and 300 MPa with inlet temperatures of 30 and 40 degrees C. UHPH-treated samples were compared to high-pasteurized milk (PA; 90 degrees C, 15 s). Results showed that all FFA increased significantly during storage only in 200 MPa samples. Lipid oxidation was measured as an accumulation of lipid hydroperoxides as the primary oxidation product and malondialdehyde and hexanal as the secondary oxidation products. Samples treated at 300 MPa presented higher malondialdehyde and hexanal content compared to 200 MPa treated-samples and to PA milk.     

New lipophilic tyrosyl esters. Comparative antioxidant evaluation with hydroxytyrosyl esters

New lipophilic esters of tyrosol, a naturally occurring phenol with interesting biological properties, have been synthesized in good yields by a chemoselective procedure, using lipase from Candida antarctica or p-toluenesulfonic acid as catalysts. Their antioxidant activities have been evaluated by the Rancimat test in lipophilic food matrices, as well as by FRAP and ABTS assays in methanolic solutions, and compared with those of previously synthesized hydroxytyrosyl esters. Free tyrosol, hydroxytyrosol, butylhydroxytoluene, and alpha-tocopherol were used as standards. All methods used for the antioxidant activity evaluation emphasized the high influence of the ortho-diphenolic structure on the antioxidant capacity, tyrosol and its derivatives being less active than hydroxytyrosol and its analogues and even less than BHT and alpha-tocopherol. In addition, the Rancimat test revealed a lower activity for ester derivatives than for their respective reference compounds (HTy or Ty), in agreement with the polar paradox. On the other hand, FRAP and ABTS methods reported an opposite behavior between the synthetic esters and their respective references. Thus, hydroxytyrosyl esters were more active than HTy, whereas tyrosyl esters were less active than Ty. The length and nature of the acyl side chain did not seem to play an important role in the antioxidant activity of either the hydroxytyrosyl or tyrosyl ester series, since no significant differences were observed among them.     

Oestrus Synchronization Treatment Induces Histomorphological Changes on the Uterine Tube Epithelium of the Gilt

The aim of this study was to determine the histomorphological changes that occurred in response to two treatments for oestrus synchronization in three different regions of the gilt's uterine tubes epithelium: the ampulla (AMP), ampulla–isthmic junction (AIJ) and isthmus (IST). Nine prepuberal gilts were divided into three groups (n = 3): (1) eCG 400 IU and hCG 200 IU (eCG/hCG), (2) progesterone agonist (P4) and (3) control group. The number of secretory cells (stained with periodic acid–Schiff reaction or PAS‐positive cells) decreased in the AMP in the P4 treated group when compared to the control group, whereas, no difference was observed in the number of PAS‐negative cells in the AMP of the three groups. A significant decrease in the number of PAS‐positive cells was observed in the AIJ and IST of the P4 treated group when compared to the eCG/hCG and control groups. An increase in the number of PAS‐negative cells was observed in the AIJ and IST in the P4 treated group. The epithelium height in the AMP and AIJ was increased in the eCG/hCG group when compared to the control and P4 groups. In this last group, we observed a reduced height compared with the other two groups for the AIJ. In the IST, there were no significant changes in the epithelium height of the control or the other two groups (eCG/hCG and P4). The epithelial cells of the P4 treated group had the least amount of cytoplasmic granules and the lowest intensity of PAS staining in the AMP, AIJ and IST. Animals treated with eCG/hCG showed an intermediate number of cytoplasmic granules and intensity in all regions evaluated. These data show that P4 treatment for synchronization induces a significant (P < 0.001) decrease of PAS‐positive cells and staining intensity of cytoplasmic granules in the different regions studied and an increased number of PAS‐negative cells in the AIJ and IST epithelium. Moreover, eCG/hCG treatment increased the height of the epithelium in the AMP and AIJ, while in this last region, the P4 treatment decreased the epithelium height. These results show that synchronization treatments with P4 and in a smaller proportion with eCG/hCG can modify the amount of PAS‐positive and PAS‐negative cells, and the epithelium height. This has influence in the secretory activity of the epithelium and possibly alters the fluid microenvironment of the gilt's uterine tube. The biological impact of regional variations in the epithelial cells of the gilt′s uterine tube needs further investigation to understand the implications that the reproductive processes can have in the uterine tube.     

Pinus chiapensis, a keystone species: Genetics,ecology, and conservation

We present an overview of recent studies carried out on Pinus chiapensis (Mart.) Andresen (Pinus strobus var. chiapensis Mart.) and provide management and conservation recommendations. Because of its wood quality, and being commonly used by 12 ethnic groups, this pine is an outstanding forest resource at mid-altitude humid mountains of southern Mexico and Guatemala. P. chiapensis appears to be a distinctive species, closely related with North American white pines, and a potential valuable resource for establishing breeding programs with such species. P. chiapensis is the most abundant tree species in early successional stands of the tropical montane cloud forest playing a key role in ecosystem regeneration particularly in areas managed under slash-and-burn practices. However, many natural stands of this pine are severely reduced. Molecular studies based on isozymes and DNA markers reveal low genetic diversity, the lowest compared with its closest relatives (P. ayacahuite, P. monticola and P. strobus). Heterozygosity and seed viability increase significantly with population size, and inbreeding depression appears to significantly decrease seed viability, suggesting the involvement of genetic factors on population decline. Low population size is associated with both lack of perturbation in well-preserved habitats and high deforestation rates in severely disturbed habitats. Conservation and management practices require preserving and restoring connections between suitable habitats to enhance gene flow between populations, and on careful programs that monitor and control slash-and-burn practices. Restoration practices should use seeds from as many tree sources as possible to reduce inbreeding risks. Spline climate models predict significant increases in temperature, decreases in precipitation and consequently an increase of aridity along the range of P. chiapensis. Thus, assisted migration would be needed to match present genotypes to forecasted climate changes.     

A new method for the determination of carbonyl compounds in wines by headspace solid-phase microextraction coupled to gas chromatography-ion trap mass spectrometry

A new analytical method for the determination of 18 carbonyl compounds [2,3-pentadione, hexanal, (E)-2-hexen-1-al, octanal, acetoin, (E)-2-octenal, furfural, decanal, (E)-2-nonenal, benzaldehyde, 5-methylfurfural, (E,E)-2-cis-6-nonadienal, β-damascenone, phenylacetaldehyde, acetophenone, (E,E)-2,4-decadienal, benzophenone, and vanillin] in wines using automated headspace solid-phase microextraction (HS/SPME) coupled to gas chromatography-ion trap mass spectrometry (GC-ITMS) was developed. Five fibers with different polarities were tested, and a study of the influence of various factors such as time and extraction temperature, desorption time and temperature, pH, and ionic strength and content in tannins, anthocyans, sucrose, SO(2), and alcoholic degree was conducted. These factors were optimized using a synthetic wine doped with the different analytes. The proposed method affords wide ranges of linearity, good linearity (r(2) > 0.998), values of repeatability and reproducibility lower than 5.5% of RSD, and detection limits ranging from 0.62 μg/L for β-damascenone to 129.2 μg/L for acetoin. Therefore, the optimized method was applied to the quantitative analysis of the aforementioned analytes in real samples of wines.     

Inhibition of morphine tolerance and dependence by the NMDA receptor antagonist MK-801.

The N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor is an important mediator of several forms of neural and behavioral plasticity. The present studies examined whether NMDA receptors might be involved in the development of opiate tolerance and dependence, two examples of behavioral plasticity. The noncompetitive NMDA receptor antagonist MK-801 attenuated the development of tolerance to the analgesic effect of morphine without affecting acute morphine analgesia. In addition, MK-801 attenuated the development of morphine dependence as assessed by naloxone-precipitated withdrawal. These results suggest that NMDA receptors may be important in the development of opiate tolerance and dependence.