The Surfaces of the Ceratonia siliqua L. (Carob) Leaflet: Insights from Physics and Chemistry.

The production of superhydrophobic coatings inspired by the surface of plant leaves is a challenging goal. Such coatings hold a bright technological future in niche markets of the aeronautical, space, naval, building, automobile, and biomedical sectors. This work is focused on the adaxial (top) and abaxial (bottom) surfaces of the leaflet of the Ceratonia silique L. (carob), a high-commercial-value Mediterranean tree cultivated in many regions of the world. The adaxial and abaxial surfaces feature hydrophobic and superhydrophobic behaviors, respectively. Their chemical composition is very simple: monopalmitin ester and palmitic acid are protuberant in the epicuticular and intracuticular wax layers of the adaxial surface, respectively, whereas 1-octacosanol dominates in the abaxial wax layers. In both surfaces, epicuticular wax is organized along a randomly oriented and intricate network of nanometer-thick and micrometer-long plates, whose density and degree of interconnection are significantly higher in the abaxial surface. The measured tilting angles for the abaxial surface (12-70°) reveal unusual variable density and water adhesion of the nanostructured plate-based texture. Optical measurements demonstrate that light reflectance/absorbance of the glaucous abaxial surface is significantly higher/lower than that of the nonglaucous adaxial surface. In both surfaces, diffuse reflectance is dominant, and the absorbance is weakly dependent on the light incidence angle. We show that the highly dense nanostructured platelike texture of the epicuticular abaxial layer of the C. siliqua leaflet works as a sophisticated light and water management system: it reflects solar radiation diffusely to lower the surface temperature, and it has superhydrophobic character to keep the surface dry. Such attributes enable efficient gas exchange (photosynthesis and respiration), transpiration, and evaporation. To mimic for the first time the abaxial surface, a templation approach was adopted using poly(dimethylsiloxane) (PDMS)/poly(methylphenylsiloxane) (PMPS) positive/negative replicas and a soft polymer/siloxane negative replica produced by the sol-gel process. Because high topographical variations of the biotemplate and wax adhesion to the biohybrid film affected the replication quality, the reproduction of the wax texture via the synthesis of 1-octacosanol-grafted siloxane-based hybrid materials is proposed as a suitable route to duplicate the abaxial surface with high fidelity. The natural chemical/physical strategy adopted by the C. siliqua leaflet to face the harsh Mediterranean climate is a powerful source of bioinspiration for the development of diffuse reflecting and superhydrophobic material systems with foreseen applications as dual-functional antiglare and water-repelling coatings.

Phytochemical characterization of Borago officinalis L. and Centaurea cyanus L. during flower development.

The present work details the nutritional and chemical compositions of borage and centaurea, at three flowering stages. Water was the main constituent, followed by total dietary fiber. Both flowers showed statistically different (p < 0.05) nutritional and chemical profiles, although in both, polyunsaturated fatty acids (PUFAs) (mainly linoleic and α-linolenic acids), free sugars (3.9-28.9% dw as fructose, glucose, and sucrose), tocopherols (with the major contribution of α-tocopherol from 1.24 to 2.75 mg/100 g dw), carotenoids (0.2-181.4 mg/100 g dw, mainly as lutein), and organic acids (6.1-14.4 g/100 g dw, mainly malic, succinic, and citric acids) were quantified. Concerning flowering, significant differences (p < 0.05) were found for some components, particularly carotenoids; however, no specific trend was observed in either of the two flower species. Thus, the present study shows that each flower species, as well as their flowering stages, may have different phytochemical and nutritional compositions.

Olive cultivar and maturation process on the oviposition preference of Bactrocera oleae (Rossi) (Diptera: Tephritidae).

The olive fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), is a key-pest in the main olives producing areas worldwide, and displays distinct preference to different olive cultivars. The present work intended to study oviposition preference towards three Portuguese cultivars (Cobrançosa, Madural, and Verdeal Transmontana) at different maturation indexes. Multiple oviposition bioassays (multiple-choice and no-choice) were conducted to assess cultivar preference. No-choice bioassays were conducted to assess the influence of different maturation indexes (MI 2; MI 3, and MI 4) in single cultivars. The longevity of olive fly adults according to the cultivar in which its larvae developed was also evaluated through survival assays.Cultivar and maturation are crucial aspects in olive fly preference. Field and laboratory assays revealed a preference towards cv. Verdeal Transmontana olives and a lower susceptibility to cv. Cobrançosa olives. A higher preference was observed for olives at MI 2 and MI 3. The slower maturation process in cv. Verdeal Transmontana (still green while the other cultivars are reddish or at black stage) seems to have an attractive effect on olive fly females, thus increasing its infestation levels. Olive fly adults from both sexes live longer if emerged from pupae developed from cv. Verdeal Transmontana fruits and live less if emerged from cv. Cobrançosa. Therefore, olive cultivar and maturation process are crucial aspects in olive fly preference, also influencing the longevity of adults.

Fatty acids derived from a food frequency questionnaire and measured in the erythrocyte membrane in relation to adiponectin and leptin concentrations.

BACKGROUND/OBJECTIVES: Evidence on the association between fatty acids and adiponectin and leptin concentrations is scarce and inconsistent, which may in part be due to limitations of dietary reporting methods. We aimed to estimate the association of fatty acids, derived from a food frequency questionnaire (FFQ) and measured in the erythrocyte membrane, with adiponectin and leptin concentrations. SUBJECTS/METHODS: We studied 330 non-institutionalized inhabitants of Porto (52.4% women; age range: 26-64 years) evaluated in 2010-2011, as part of the EPIPorto cohort study. Fatty acids were derived from a validated semiquantitative FFQ and measured in the erythrocyte membrane by gas chromatography. Serum concentrations of adiponectin and leptin were determined through radioimmunoassay. Regression coefficients (ß) and 95% confidence intervals (95% CI) were obtained from linear regression models, after controlling for gender, age, education, leisure time physical activity and total body fat percentage (obtained from dual energy X-ray absorptiometry). RESULTS: Fatty acids measured by FFQ showed no significant associations with both adipokines. Lauric and linoleic acids, measured in the erythrocyte membrane, were significantly and positively associated with adiponectin (ß=0.292, 95% CI: 0.168-0.416; ß=0.150, 95% CI: 0.020-0.280) and leptin (ß=0.071, 95% CI: 0.003-0.138; ß=0.071, 95% CI: 0.002-0.140), whereas total n-3, eicosapentaenoic and docosahexaenoic acids were significantly but negatively associated with adiponectin (ß=-0.289, 95% CI: -0.420 to -0.159; ß=-0.174, 95% CI -0.307 to -0.040; ß=-0.253, 95% CI -0.383 to -0.124) and leptin (ß=-0.151, 95% CI: -0.220 to -0.083; ß=-0.080, 95% CI: -0.151 to -0.009; ß=-0.146, 95% CI: -0.214 to -0.078). Positive significant associations of palmitic and trans-fatty acids with adiponectin were also observed. CONCLUSIONS: A positive association of lauric and linoleic acids and a negative association of total n-3 fatty acids with both adipokines were observed only with fatty acids measured in the erythrocyte membrane.

Commercial squids: characterization, assessment of potential health benefits/risks and discrimination based on mineral, lipid and vitamin E concentrations.

The most consumed squid species worldwide were characterized regarding their concentrations of minerals, fatty acids, cholesterol and vitamin E. Interspecific comparisons were assessed among species and geographical origin. The health benefits derived from squid consumption were assessed based on daily minerals intake and on nutritional lipid quality indexes. Squids contribute significantly to daily intake of several macro (Na, K, Mg and P) and micronutrients (Cu, Zn and Ni). Despite their low fat concentration, they are rich in long-chain omega-3 fatty acids, particularly docosahexaenoic (DHA) and eicosapentanoic (EPA) acids, with highly favorable ω-3/ω-6 ratios (from 5.7 to 17.7), reducing the significance of their high cholesterol concentration (140-549 mg/100g ww). Assessment of potential health risks based on minerals intake, non-carcinogenic and carcinogenic risks indicated that Loligo gahi (from Atlantic Ocean), Loligo opalescens (from Pacific Ocean) and Loligo duvaucelii (from Indic Ocean) should be eaten with moderation due to the high concentrations of Cu and/or Cd. Canonical discriminant analysis identified the major fatty acids (C14:0, C18:0, C18:1, C18:3ω-3, C20:4ω-6 and C22:5ω-6), P, K, Cu and vitamin E as chemical discriminators for the selected species. These elements and compounds exhibited the potential to prove authenticity of the commercially relevant squid species.

Blood antioxidant and oxidative stress biomarkers acute responses to a 1000-m kayak sprint in elite male kayakers.

AIM: This study aimed to investigate the response of blood antioxidants and biomarkers of lipid peroxidation, muscle damage and inflammation to a 1000m kayak trial in elite male kayakers. METHODS: Enzymatic (superoxide dismutase [SOD], glutathione reductase [Gr] and glutathione peroxidase [GPx] activities) and non-enzymatic (total antioxidant status [TAS], uric acid, α-tocopherol, α-carotene, ß-carotene, lycopene and lutein and zeaxanthin) antioxidants, thiobarbituric acid reactive substances (TBARS), creatine kinase (CK), interleukin-6 (IL-6) and cortisol were determined in 15 elite male kayakers before and 15 min after a 1000-m kayak simulated race. RESULTS: Both enzymatic and non-enzymatic antioxidants were unaffected by exercise, with the exception of α-carotene which decreased (P=0.013). Uric acid levels were incremented following exercise (P=0.016). The acute exercise resulted in a significant decrease in TAS (P=0.001) and in an increase in CK (P=0.023), TBARS (P<0.001) and IL-6 (P=0.028). CONCLUSION: Our study suggests that a 1000-m kayak simulated race induces oxidative stress and damage in highly-trained kayakers.

Fat-soluble vitamin (A, D, E, and ß-carotene) contents from a Portuguese autochthonous cow breed--Minhota.

In Portugal, commercial milk is obtained almost entirely from cows of the Holstein breed. However, other breeds may also show dairy aptitude, and produce milk of good quality. The aim of this study was to investigate the effect of the breed factor in the vitamin contents of milks from Holstein and the autochthonous Portuguese breed Minhota, as well as possible variations due to season. Milk samples were collected from 15 cows of each breed. Milk from Minhota cows showed higher contents of retinol, retinyl palmitate, α-tocopherol, and ß-carotene. No differences were verified between breeds regarding vitamin D(3) content, but Holstein cows presented higher amounts of provitamin D(3). Seasonal variations were not observed for milk samples from Holsteins, and only a very small variation was observed for milk samples from Minhota cows, mainly in retinol, α-tocopherol, and provitamin D(3) levels. This study revealed that breed has a pronounced effect on milk fat-soluble vitamins, which is an important observation regarding future genetic selection plans. Also, the increased vitamin amounts found in milk from Minhota cows can be regarded as an important nutritional parameter that can potentiate the economical value of this breed.

Fatty acid profile of human milk of Portuguese lactating women: prospective study from the 1st to the 16th week of lactation.

BACKGROUND/AIMS: Fatty acid (FA) composition varies over the course of the day and during lactation. The aim of this study was to evaluate FA composition and its compositional stability in human milk, from day 7 to week 16 of lactation. METHODS: Human milk was collected from all feedings over 24 h at day 7 and weeks 4, 8, 12 and 16 of lactation in 31 lactating women. FAs were analyzed through gas chromatography. Comparisons were made with analysis of variance. RESULTS: Total monounsaturated FAs decreased from 33.04 +/- 2.58% wt/wt at day 7 to 31.48 +/- 3.32% wt/wt at week 16 of lactation, much at the expenses of the decrease in the major monounsaturated FA found in human milk, oleic acid. Main polyunsaturated FAs n-6 and n-3 showed fluctuations from day 7 up to week 16 of lactation, but with no statistical significance. Arachidonic acid significantly decreased from transitional to mature milk. CONCLUSIONS: The FA profile obtained throughout the study time points presented very low levels of oleic acid and very high linoleic acid/alpha-linoleic acid ratios which reflect recent changes in Portuguese women's food patterns. Despite this, the arachidonic acid/docosahexaenoic acid ratio [corrected] remained constant during the study, suggesting a protective metabolic mechanism.

Analysis of heterocyclic aromatic amines in foods by gas chromatography-mass spectrometry as their tert.-butyldimethylsilyl derivatives.

A derivatization method for the analysis of 12 heterocyclic aromatic amines (HAs) in food, by gas chromatography-electron impact mass spectrometry, was developed. The amines are derivatized in a one-step reaction with N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide. The derivatives are characterized by easy-to-interpret mass spectra due to the prominent ion [M-57]+ by loss of a tert.-butyldimethylsilyl group, allowing quantification in the selected-ion monitoring mode at the picogram level. The effect of temperature, time, and reagents on the formation of the derivatives was monitored in detail. Quality parameters were evaluated in the optimum working conditions. This derivatization method is not applicable to the pyridoimidazoles Glu-P-1 and Glu-P-2 and to the beta-carboline harman due to incompletely derivatization. The instability of the imidazolquinoline and imidazoquinoxaline derivatives, requiring their injection on the same working day, is a further drawback. This simple, rapid and accurate derivatization procedure is suitable for routine analysis, as illustrated by the analysis of some common foods.

Gas chromatographic-mass spectrometric quantification of 4-(5-)methylimidazole in roasted coffee after ion-pair extraction.

A GC-MS method is described for quantification of 4-(5-)methylimidazole (4MI) in coffee. Although tested, GC-flame ionization detection proved inadequate for this purpose due to the complexity of the coffee matrix. The developed method was based on ion-pair extraction with bis-2-ethylhexylphosphate and derivatization with isobutylchloroformate. Quantification was carried out by the standard addition method using 2-ethylimidazole as internal standard. Reproducibility data from the complete procedure are presented. Mean recoveries were higher than 98%. The method was applied to green and roasted coffee samples from the two most important varieties, arabica and robusta, and to commercial "torrefacto" coffee blends. 4MI was not detected in the green coffee samples analysed and ranged from 0.307 to 1.241 mg/kg in roasted samples.

Influence of olive storage period on oil quality of three Portuguese cultivars of Olea europea, Cobrançosa, Madural, and Verdeal Transmontana.

Olives (Olea europaea cv. Cobrançosa, Madural, and Verdeal Transmontana) used for oil production were stored, in plastic containers, at 5 +/- 2 degrees C (70% relative humidity) for three different periods before oil extraction: 0, 7, and 14 days (T(0), T(7), and T(14), respectively). In the crop year 1997/1998 this procedure was done only for cv. Cobrançosa and in 1998/1999 for the three cultivars. After storage, the oils were extracted from the fruits, and the acidity, peroxide value, coefficients of specific extinction at 232 and 270 nm, stability, color, p-anisidine value, fatty acids, and tocopherols compositions were determined. The results confirm that storage of fruits produces losses in the olive oil quality. Acidity and stability to oxidation indicate a progressive deterioration of oil quality as fruit is stored. The storage time affects the total tocopherols contents, namely, alpha-tocopherol, which clearly decreased during fruit storage. The oil quality of the Verdeal Transmontana cultivar deteriorated more rapidly than that of Cobrançosa and Madural cultivars. This study also shows that cv. Cobrançosa, the main cultivar in the region, is a good choice in terms of final olive oil quality.

Simultaneous determination of retinol, beta-carotene and alpha-tocopherol in adipose tissue by high-performance liquid chromatography.

A method is described for evaluation of fat-soluble vitamin in human adipose tissue with the aim to obtain, accurately and within the shortest analysis time, a time-integrated measure of exposure to vitamins from the diet. Fat tissue was deproteinized with ethanol and extracted with n-hexane. Normal-phase HPLC was performed in a Lichrosorb Si60 column with a gradient of n-hexane-2-propanol at 1 ml/min. Detection was accomplished using a diode-array system (for retinol and beta-carotene) in series with a fluorescence detector (alpha-tocopherol). The method was validated and applied to human adipose tissue in a total of 140 subjects. The mean contents found were 0.43, 0.84, 240.3 microg/g for retinol, beta-carotene and alpha-tocopherol, respectively. The method is sensitive enough for detecting the compounds in 1.6 mg of adipose tissue considering the lowest concentration found.

Discriminate analysis of roasted coffee varieties for trigonelline, nicotinic acid, and caffeine content.

Arabica and robusta roasted coffees from several geographical origins, in a total of 29 samples, were characterized for their contents in caffeine, trigonelline, and nicotinic acid by a recently developed HPLC/diode-array detector method. All samples were subjected to the same roasting procedure in order to eliminate the variations due to this process. Characterization was achieved by applying multivariate and nonparametric analysis to the chromatographic results. The two coffee varieties were clearly separated by their trigonelline and caffeine contents. Nicotinic acid could not be used as a variety discriminate factor. There was no association with the geographical origin of the samples.

In vivo induction of a high-avidity, high-frequency cytotoxic T-lymphocyte response is associated with antiviral protective immunity.

Many approaches are currently being developed to deliver exogenous antigen into the major histocompatibility complex class I-restricted antigen pathway, leading to in vivo priming of CD8(+) cytotoxic T cells. One attractive possibility consists of targeting the antigen to phagocytic or macropinocytic antigen-presenting cells. In this study, we demonstrate that strong CD8(+) class I-restricted cytotoxic responses are induced upon intraperitoneal immunization of mice with different peptides, characterized as CD8(+) T-cell epitopes, bound to 1-microm synthetic latex microspheres and injected in the absence of adjuvant. The cytotoxic response induced against a lymphocytic choriomeningitis virus (LCMV) peptide linked to these microspheres was compared to the cytotoxic T-lymphocyte (CTL) response obtained upon immunization with the nonreplicative porcine parvovirus-like particles (PPV:VLP) carrying the same peptide (PPV:VLP-LCMV) previously described (C. Sedlik, M. F. Saron, J. Sarraseca, I. Casal, and C. Leclerc, Proc. Natl. Acad. Sci. USA 94:7503-7508, 1997). We show that the induction of specific CTL activity by peptides bound to microspheres requires CD4(+) T-cell help in contrast to the CTL response obtained with the peptide delivered by viral pseudoparticles. Furthermore, PPV:VLP are 100-fold more efficient than microspheres in generating a strong CTL response characterized by a high frequency of specific T cells of high avidity. Moreover, PPV:VLP-LCMV are able to protect mice against a lethal LCMV challenge whereas microspheres carrying the LCMV epitope fail to confer such protection. This study demonstrates the crucial involvement of the frequency and avidity of CTLs in conferring antiviral protective immunity and highlights the importance of considering these parameters when developing new vaccine strategies.

Gas chromatographic quantification of amino acid enantiomers in food matrices by their N(O,S)-ethoxycarbonyl heptafluorobutyl ester derivatives.

Several amino acid enantiomer derivatives were prepared with different chloroformates and analysed by gas chromatography (GC) on a Chirasil-L-Val GC column, at a temperature below 200 degrees C. Among them the N(O,S)-ethoxycarbonyl heptafluorobutyl esters presented the best compromise between short retention times, high yield responses and good resolution for almost all the tested amino acids. These derivatives proved to be suited for quantification of amino acids in aqueous media, with L-p-chlorophenylalanine as internal standard. The developed procedure was applied to several food samples for determination of their free amino acid profiles.

Maternal education and maternal responsiveness as predictors of play competence in low birth weight, premature infants: a preliminary report.

Videotaped play interactions of 182 low birth weight, premature toddlers and their mothers were used to investigate whether a model including maternal responsiveness together with level of maternal education was a better predictor of play maturity scores of the children than a model consisting of education level alone. The videotapes were coded using the responsiveness subscale of the Maternal Behavior Rating Scale. Regression analyses revealed that maternal education alone significantly predicted play scores, accounting for 9% of the variance. However, the model that included both maternal responsiveness and maternal education together also significantly predicted play scores and accounted for 20% of the variance. Including responsiveness together with maternal education significantly increased the proportion of variance accounted for by maternal education alone. Implications of these findings are discussed.