Isolation, physicochemical properties, and the macromolecular composition of the vitelline and fertilization envelopes from Xenopus laevis eggs

As a step toward defining in molecular terms the sperm-triggered block to polyspermy reaction established by the egg at fertilization, vitelline (VE) and fertilization (FE) envelopes were isolated from eggs of the Sounth African clawed toad Xenopus laevis and some of their physicochemical properties determined. Envelopes were isolated after lysis of the fertilized or unfertilized eggs by sieving techniques; isolated envelopes retained their in situ morphology as determined by electron microscopy. The isolated envelopes had different solubility properties and, in general, VE was more readily dissolved by aqueous solvents than FE, although both could be completely dissolved by detergents or chaotropic agents. Changes in envelope solubility correlated with the progression of the cortical reaction implicating a role for cortical granule material in modifying the solubility properties of the envelope. The VE and FE were composed of protein and carbohydrate with no lipid components detected. As determined by immunodiffusion experiments, the FE contained the same antigens as the VE plus components derived from the cortical granules and the innermost jelly layer, J. The macromolecular composition of the envelopes was determined by sodium dodecyl sulfate gel electrophoresis. The VE contained at least 11 glycoproteins with molecular weights ranging from 125 000 to less than 16 000 with two components (40 000 and 33 000) accounting for almost two-thirds of the total stainable material. The FE contained ten glycoproteins that had the same molecular weights as those in the VE. One glycoprotein component underwent a reduction in molecular weight from 77 000 to 67 500 when the VE was converted to the FE. This molecular weight change was interpreted as the probable result of limited proteolysis. In addition, the FE gel electrophoresis patterns contained macromolecular components derived from the cortical granules and jelly layer, J, consistent with the immunodiffusion experiments. These components were absent when the FE was prepared in the absence of Ca2+, suggesting a role for Ca2+ in binding the VE, cortical granules, and J components together. We concluded that the conversion of the glycoproteinaceous VE to FE at fertilization is caused by interaction of the VE with components from the cortical granules and jelly layer J. These interactions are of both a chemical and physical nature.     

Cranial cruciate ligament repair in a calf

A 1-month-old calf was unable to use its right pelvic limb because of rupture of the cranial cruciate ligament. The ligament was replaced with a wedge of bone from the cranial one-third of the patella and the attached portion of the middle patellar tendon. Postoperative complications limited function of the joint.     

Inhibition of Bacterial Pathogens in Medium and on Spinach Leaf Surfaces using Plant‐Derived Antimicrobials Loaded in Surfactant Micelles

Encapsulation of hydrophobic plant essential oil components (EOC) into surfactant micelles can assist the decontamination of fresh produce surfaces from bacterial pathogens during postharvest washing. Loading of eugenol and carvacrol into surfactant micelles of polysorbate 20 (Tween 20), Surfynol® 485W, sodium dodecyl sulfate (SDS), and CytoGuard® LA 20 (CG20) was determined by identification of the EOC/surfactant‐specific maximum additive concentration (MAC). Rheological behavior of dilute EOC‐containing micelles was then tested to determine micelle tolerance to shearing. Antimicrobial efficacy of EOC micelles against Escherichia coli O157:H7 and Salmonella enterica serotype Saintpaul was first evaluated by the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Pathogen‐inoculated spinach was treated with eugenol‐containing micelles applied via spraying or immersion methods. SDS micelles produced the highest MACs for EOCs, while Tween 20 loaded the lowest amount of EOCs. Micelles demonstrated Newtonian behavior in response to shearing. SDS and CG20‐derived micelles containing EOCs produced the lowest MICs and MBCs for pathogens. E. coli O157:H7 and S. Saintpaul were reduced on spinach surfaces by application of eugenol micelles, though no differences in numbers of surviving pathogens were observed when methods of antimicrobial micelle application (spraying, immersion) was compared (P ≥ 0.05). Data suggest eugenol in SDS and CG20 micelles may be useful for produce surface decontamination from bacterial pathogens during postharvest washing.     

Protective effects of phenolic constituents from Cytisus multiflorus,Lamium album L. and Thymus citriodorus on liver cells

The present study investigated the antioxidant and cytoprotective effects of purified ethanolic extracts of Cytisus multiflorus, Lamium album L. and Thymus citriodorus plants. These extracts showed high antioxidant activity in DPPH and reducing power assays. Using a model of chemical stress induced by potassium dichromate (DK) in human hepatoblastoma HepG2 cells, 50 μg/mL of C. multiflorus, L. album and T. citriodorus extracts decreased the rate of reactive oxygen species (ROS) production by 35%, 26% and 20%, respectively, when exposed to 25 μM of DK. This effect was also observed for the treatment of cells with individual polyphenolic compounds determined in the extracts, or with mixtures prepared with individual polyphenolic compounds simulating the phenolic composition of the extracts. Additionally, the purified ethanolic extracts and the prepared polyphenolic mixtures showed a cytoprotective effect against DK-induced toxicity. The overall results emphasize the contribution of polyphenols in antioxidant and cytoprotective properties of the studied plants.     

Energy‐dependent effects of resveratrol in Saccharomyces cerevisiae

The metabolic effects induced by resveratrol have been associated mainly with the consumption of high‐calorie diets; however, its effects with standard or low‐calorie diets remain unclear. To better understand the interactions between resveratrol and cellular energy levels, we used Saccharomyces cerevisiae as a model. Herein it is shown that resveratrol: (a) decreased cell viability in an energy‐dependent manner; (b) lessening of cell viability occurred specifically when cells were under cellular respiration; and (c) inhibition of oxygen consumption in state 4 occurred at low and standard energy levels, whereas at high energy levels oxygen consumption was promoted. These findings indicate that the effects of resveratrol are dependent on the cellular energy status and linked to metabolic respiration. Importantly, our study also revealed that S. cerevisiae is a suitable and useful model to elucidate the molecular targets of resveratrol under different nutritional statuses. Copyright © 2016 John Wiley & Sons, Ltd.     

Antioxidant properties,radical scavenging activity and biomolecule protection capacity of flavonoid naringenin and its glycoside naringin: a comparative study

BACKGROUND: This study was designed to evaluate and compare antioxidant capacity and radical scavenging activity of naringin and its aglycone by different in vitro assays. The effects of flavanones on lipid peroxidation, glutathione (GSH) oxidation and DNA cleavage were also assessed. RESULTS: The results showed that naringenin exhibited higher antioxidant capacity and hydroxyl and superoxide radical scavenger efficiency than naringin. Our results evidenced that glycosylation attenuated the efficiency in inhibiting the enzyme xanthine oxidase and the aglycone could act like a more active chelator of metallic ions than the glycoside. Additionally, naringenin showed a greater effectiveness in the protection against oxidative damage to lipids in a dose‐dependent manner. Both flavanones were equally effective in reducing DNA damage. However, they show no protective effect on oxidation of GSH. CONCLUSION: The data obtained support the importance of characterizing the ratio naringin/naringenin in foods when they are evaluated for their health benefits. Copyright © 2010 Society of Chemical Industry     

The role of K+ and H+ transport systems during glucose‐ and H2O2‐induced cell death in Saccharomyces cerevisiae

Glucose, in the absence of additional nutrients, induces programmed cell death in yeast. This phenomenon is independent of yeast metacaspase (Mca1/Yca1) and of calcineurin, requires ROS production and it is concomitant with loss of cellular K⁺ and vacuolar collapse. K⁺ is a key nutrient protecting the cells and this effect depends on the Trk1 uptake system and is associated with reduced ROS production. Mutants with decreased activity of plasma membrane H⁺‐ATPase are more tolerant to glucose‐induced cell death and exhibit less ROS production. A triple mutant ena1‐4 tok1 nha1, devoid of K⁺ efflux systems, is more tolerant to both glucose‐ and H₂O₂‐induced cell death. We hypothesize that ROS production, activated by glucose and H⁺‐ATPase and inhibited by K⁺ uptake, triggers leakage of K⁺, a process favoured by K⁺ efflux systems. Loss of cytosolic K⁺ probably causes osmotic lysis of vacuoles. The nature of the ROS‐producing system sensitive to K⁺ and H⁺ transport is unknown. Copyright © 2010 John Wiley & Sons, Ltd.     

Authentication of Green Asparagus Varieties by Near-Infrared Reflectance Spectroscopy

ABSTRACT: Near-infrared reflectance spectroscopy (NIRS) was used for the authentication of 2 green asparagus varieties (Taxara and UC-157), grown in Huetor-Tajar (Granada, Spain) protected by the Quality Specific Appellation "Espárragos de Huétor-Tájar". To develop the prediction model, the method chosen was modified partial least square (MPLS) regression. Two sample sets (N = 219 and N2 = 145 samples, respectively) were used to obtain the calibration equations. The standard error of cross-validation (SECV) and the r2 value were 0.082 and 0.97, respectively, for the 1st calibration set and 0.077 and 0.97 for the 2nd calibration set. The 2nd chemometric model obtained was tested with independent validation sample set (N3 = 74 samples), and the resulting values for standard error of prediction (SEP) and for r2 were 0.07 and 0.96, respectively. These results prove that NIRS is an accurate technology for identification and authentication of asparagus varieties and easily implemented in industry.     

Use of immunological techniques for detecting species substitution in raw and smoked fish

 Immunological techniques based on double immunodiffusion and immunodotting have been designed to detect the substitution of halibut (Hippoglossus hippoglossus) for sole (Solea solea) fillets. An immunodotting technique has been also developed to differentiate between smoked cod (Gadus morhua) and smoked eel (Anguilla anguilla) fillets. Antisera raised against water-soluble extracts of raw halibut and smoked cod were employed. Antiserum to cod proteins did not show cross-reaction with eel proteins, whereas antiserum to halibut proteins cross-reacted with sole proteins. Species-specific antiserum to halibut proteins was achieved by adsorption on a polymer of sole proteins. These methods are very easy to perform and provide the basis for the development of rapid tests for detecting species substitution in fish products. Received: 18 April 1996/Revised version: 5 July 1996     

Structure and degree of polymerisation of fructooligosaccharides present in roots and leaves of Stevia rebaudiana (Bert.) Bertoni

Fructooligosaccharides have been isolated from roots and leaves of Stevia rebaudiana, by hot aqueous extraction, followed by precipitation with ethanol. Their structure has been determined using methylation and NMR analysis, MALDI-TOF, and ESI-MS. Fructooligosaccharides contained almost exclusively (2→1)-linked β-fructofuranosyl, with terminal α-glucopyranosyl and β-fructofuranosyl units. MALDI-TOF and ESI-MS analyses showed the wide range of degree of polymerisation (DP) present in various extracts. From roots and leaves, three different fractions gave profiles of homologous series, with DPs ranging up to 17 with MALDI-TOF and 19 using ESI-MS. These inulin-type fructooligosaccharides were the major component of extracts from S. rebaudiana roots and significant components from the leaves.