Effect of decorin and selected glycosylation inhibitors on the adhesion of caruncular epithelial cells of pregnant cows—part I

Adhesion process ensures the formation of the appropriate connection between mother and foetus during placentation and further placental development, which determines physiological pregnancy course. Extracellular matrix of foetal membranes are a rich source of biologically active proteins, the synthesis of which is regulated by hormones. Depending on the stage of pregnancy, the protein profile of the placenta changes, thanks to which its remodelling is possible. The aim of the study was to evaluate the effect of decorin, as well as selected glycosylation inhibitors on the adhesion of caruncular epithelial cells derived from cows during pregnancy. Placental cells were isolated from healthy, pregnant (2nd and 4th month) cows after slaughter, which allowed for the establishment of 4 primary cell cultures without visible cells of fibroblast morphology. The presence of decorin in cell monolayer and cell lysates was determined by the use of immunocytochemistry and Western blotting, respectively. The viability of cells was evaluated by MTT assay. The adhesion of cells to fibronectin was measured spectrophotometrically. Protein N-glycosylation and O-glycosylation have a modulating effect on the adhesion and viability of placental cells during early–mid pregnancy. Decorin and tunicamycin were shown to have anti-adhesive properties with respect to caruncular cells of the pregnant bovine uterus.     

Development of testis and digestive tract in longnose gar (Lepisosteus osseus) at the onset of exogenous feeding of larvae and in juveniles

The aim of this study was to describe the ontogenetic development of the testis and the alimentary tract in longnose gar (Lepisosteus osseus) related to fish size and age at the onset of exogenous feeding and late ontogenesis. Using light microscopy, testes were first detected histologically by the appearance of primordial germ cells 9 days after the first exogenous feeding [31–31.5 mm total body length(TL)] and presumptive seminiferous tubules (maleness characteristic) in fish of 107 mm TL. The present histological studies indicated that the alimentary tract of lepisosteids is completely functional at the beginning of exogenous feeding, several days before the completion of yolk absorption. Based on these results, we have concluded that garfish larvae/juveniles can be effectively trained to consume formulated diets at early stages, after an initial feeding of live food for 2–3 days (23.5 mm TL). Our findings provide evidence of the first controlled rearing of longnose gar using live and formulated diets, providing the possibility of experimental work with this non‐teleost fish.     

Effect of caffeic acid on the color of red wine

The copigmentation effect of prefermentation additions of different doses of caffeic acid was investigated during the 1997 harvest. Microfermentation with the major red grape cultivars Listán negro and Negramoll, grown in the Canary Islands, was carried out with the same protocol. Visible and UV spectra were registered periodically. HPLC chromatograms were carried out. The color enhancement of cv. Negramoll wine varied between 13 and 75% (AU at 520 nm), and that of cv. Listán negro wine between 25 and 45% at the end of fermentation. During aging these values were enhanced to reach even >100% in some cases. An initial complex of the 1:1 type, where one molecule of caffeic acid associates with one molecule of anthocyanin, has been identified using the mathematical procedure of Brouillard et al. (J. Am. Chem. Soc. 1989, 111, 2604-2610). Caffeic acid seems to contribute to color stability and protection against oxidation. The importance of nonpigment composition in pigment extraction and color retention during and after fermentation is demonstrated.     

Characterization of anthocyanins from the fruits of baguaçu (Eugenia umbelliflora Berg)

Anthocyanin pigments in the berries of bagua?u (Eugenia umbelliflora Berg), a tropical fruit from Brazil, were extracted with 0.1% HCl in ethanol, and the crude anthocyanin extract was purified by Amberlite XAD-7 open-column chromatography. Six major anthocyanins were isolated by preparative HPLC, and their chemical structures were identified by spectroscopic methods (TLC, UV-vis, MS, and (1)H NMR). Delphinidin 3-O-beta-glucopyranoside, cyanidin 3-O-beta-glucopyranoside, petunidin 3-O-beta-glucopyranoside, pelargonidin 3-O-beta-glucopyranoside, peonidin 3-O-beta-glucopyranoside, and malvidin 3-O-beta-glucopyranoside were identified. On the basis of chromatographic data the total anthocyanin content was 342 mg/100 g of fresh bagua?u berries. Therefore, the concomitant presence of six anthocyanins in a single plant species makes this product promising as a new pigment source.     

Quantification of a Sulfated Marine-Inspired Antifouling Compound in Several Aqueous Matrices: Biodegradation Studies and Leaching Assays from Polydimethylsiloxane Coatings

The development of marine-inspired compounds as non-toxic antifouling (AF) agents has been pursued in the last years. Sulfur is the third most common element in seawater. Sulfur is present in oxygenated seawater as sulfate anion (SO₄²⁻), which is the most stable combination of sulfur in seawater, and several promising AF secondary metabolites with sulfate groups have been described. However, sulfated compounds proved to be an analytical challenge to quantify by HPLC. Taking these facts into consideration, this work presents the development and validation of a method for the quantification of gallic acid persulfate (GAP) in seawater and ultrapure water matrix, based on hydrophilic interaction liquid chromatography (HILIC). This method was used to evaluate GAP stability following several abiotic and biotic degradation assays, and to quantify its release in seawater from room-temperature-vulcanizing polydimethylsiloxane commercial coating. GAP was very stable in several water matrices, even at different pH values and in the presence/absence of marine microorganisms and presented a leaching value lower than 0.5%. This work discloses HILIC as an analytical method to overcome the difficulties in quantifying sulfated compounds in water matrices and highlights the potential of GAP as a promising long-lasting coating.     

Stacking HMW-GS transgenes in bread wheat: Combining subunit 1Dy10 gives improved mixing properties and dough functionality

We have determined the technological properties of four lines containing combinations of three HMW-GS transgenes, encoding HMW-GS 1Ax1, 1Dx5 and 1Dy10. These lines were produced by conventional crossing of three single transgenic lines of the bread wheat cultivar Anza that contains the endogenous HMW-GS pairs 1Dx2 + 1Dy12 and 1Bx7* + 1By8 and is null for the Glu-A1 locus. Consequently, the total number of HMW-GS ranged from 4 in the control line Anza to 7 in line T618 which contains all three HMW-GS transgenes. The lines were studied over two years using a range of widely used grain and dough testing methods. All lines with transgenic subunits showed higher levels of glutenin proteins than the Anza control, and these differences were highly significant for lines T616, T617 and T618, containing, respectively, the transgenes encoding HMW-GS 1Ax1 and 1Dy10, 1Dx5 and 1Dy10 and 1Ax1, 1Dx5 and 1Dy10. These increases in glutenin levels are compensated by lower levels of gliadins present in transgenic lines. These changes affected the ratio of polymeric to monomeric gluten proteins (poly:mono), the ratio of HMW-GS to LMW-GS (HMW:LMW) and the contents of individual 1Ax, 1Bx, 1By, 1Dx and 1Dy subunits. Transgenic lines expressing subunit 1Dy10 together with x-type subunits (T616, T617 and T618) were superior to line T606, which had only increases in x-type subunits. In particular, the combination of transgenic subunits 1Dx5 and 1Dy10 (line T617) gave better dough rheological properties than the other combinations of transgenic subunits. For example, dough development time and stability were increased by 3.5-fold and 8.5-fold, respectively, while the mixing tolerance index (MTI) was decreased by 3.3-fold in line T617 with respect to the control line. Alveograph analyses showed that all four transgenic combinations had increased P values compared to the Anza control but subunit 1Dx5 greatly reduced the extensibility (L). These results show that stacking HMW-GS transgenes by conventional crossing is a valid strategy for the improvement of wheat quality, with different effects being related to the different HMW-GS combinations.     

Histopathological and behaviour effects of formaldehyde treatment in juvenile mullet,Mugil liza (Valenciennes)

Considerable losses may be related to diseases caused by parasites in aquaculture. Formaldehyde (37% gas dissolved in water) is an effective chemotherapeutic, widely used to control ectoparasites in fish. However, for many species of fish used in aquaculture, the control of external parasites and the deleterious effects of formaldehyde in short‐term baths are still uncertain. In order to clarify some gaps, 1 h‐prophylactic baths (67.5, 135, 270, 405 and 540 mg L^?1 formaldehyde in triplicate) were performed in juvenile mullet, Mugil liza (Valenciennes). Results showed that prophylactic baths of 135 mg L^?1 formaldehyde by 1 h did not cause death in tested mullets. At this concentration, histopathologies were mild and fish survival was high. However, fish exposed to increasing concentrations of formaldehyde presented severe histopathologies in the gills besides behavioural and morphological alterations. The histopathological and behavioural implications of formaldehyde use in aquaculture are discussed.     

<Emphasis Type="Italic">Cornus mas</Emphasis> (L.) Fruit as a Potential Source of Natural Health-Promoting Compounds: Physico-Chemical Characterisation of Bioactive Components

Interest in new sources of anti-inflammatory and antioxidant compounds has recently become a major research issue, with the cornelian cherry (Cornus mas L.) receiving particular attention for its significant amounts of phenolic compounds and vitamins, which exhibit a wide range of biological and pharmacological properties. This study was aimed at increasing knowledge regarding the cornelian cherry in Italy through the analysis of biologically active substances in the locally available genotype “Chieri”. Spectrophotometric methods were applied to evaluate antioxidant activity, total anthocyanin content and total polyphenolic content. Identification and quantification of the main phytochemical compounds (polyphenols, monoterpenes, organic acids and vitamin C) was performed via high performance liquid chromatography coupled to a diode array detector. C. mas extracts showed high levels of total soluble solids and low acidity. High amounts of phenolic secondary metabolites were observed, with particular reference to anthocyanins (134.71 mg_C3G/100 g_FW), which confer remarkable nutraceutical properties to the analysed samples. These results highlight the potential of C. mas fruits as a good source of natural antioxidants, suggesting their use as a functional food. Future studies should focus on identifying other specific phytochemical compounds and the genetic traits of local varieties in order to improve cornelian cherry cultivars for food and medicine production.     

Protective effect of structurally diverse grape procyanidin fractions against UV-induced cell damage and death

UV radiation leads to the generation of reactive oxygen species (ROS). These molecules exert a variety of harmful effects by altering key cellular functions and may result in cell death. Several studies have demonstrated that human skin can be protected against UV radiation by using plant-derived antioxidants. Here we evaluated the in vitro capacity of several antioxidant polyphenolic fractions from grape, which differ in their degree of polymerization and percentage of galloylation, to protect HaCaT human keratinocytes against UV-induced oxidative damage. These fractions inhibited both basal and UVB- or UVA-induced intracellular ROS generation in this cell line. Consequently, the same fractions inhibited p38 and JNK1/2 activation induced by UVB or UVA radiation. The highest protective effect was for fractions rich in procyanidin oligomers and gallate esters. These encouraging in vitro results support further research and should be taken into consideration into the clinical pharmacology of plant-derived polyphenolic extracts as novel agents for skin photoprotection.