Hydroxytyrosol-rich olive mill wastewater extract protects brain cells in vitro and ex vivo

Elevated oxidative and nitrosative stress both impair the integrity and functioning of brain tissue, especially in aging. As long-term intake of plant foods rich in antioxidant phenolics, such as extra virgin olive oil, positively modulates surrogate markers of many human pathological alterations, the interest in cheap and abundant sources of such phenolics is rapidly growing. Olive mill wastewater is particularly rich in hydroxytyrosol, an o-diphenol with powerful antioxidant, anti-inflammatory, and antithrombotic activities. Due to the deleterious effect of oxidative stress on brain cell survival, the efficacy of a hydroxytyrosol-rich extract to attenuate Fe2+- and nitric oxide (NO)-induced cytotoxicity in murine-dissociated brain cells was investigated. The addition of either Fe2+ or SNP (an NO donor) caused both a severe loss of cellular ATP and a markedly depolarized mitochondrial membrane potential. Preincubation with hydroxytyrosol significantly attenuated the cytotoxic effect of both stressors, although with different efficiencies. Mice feeding studies were performed to assess the brain bioactivity of hydroxytyrosol ex vivo. Subchronic, but not acute, administration of 100 mg of hydroxytyrosol per kilogram body weight for 12 days enhanced resistance of dissociated brain cells to oxidative stress, as shown by reduced basal and stress-induced lipid peroxidation. Also, basal mitochondrial membrane potential was moderately hyperpolarized (P < 0.05), an effect suggestive of cytoprotection. In synthesis, the ex vivo data provide the first evidence of neuroprotective effects of oral hydroxytyrosol intake. Keywords: Hydroxytyrosol; olive mill wastewater; dissociated brain cells; oxidative stress; brain; Mediterranean diet.     

Evaluation of antiradical capacity by H2O2-hemin-induced luminol chemiluminescence

This paper describes a screening method for antioxidant potential determination based on luminol/hemin/hydrogen peroxide chemiluminescence. The emission depletion, caused by an antiradical compound added during the chemiluminescence decay, is proportional to the number of reactive species trapped. Therefore, the difference between the areas of the emission decay curves, obtained in the absence and in the presence of the potential antioxidant, is a measure for the antiradical capacity of the sample. The technique has been applied to measure the antiradical capacity of pure compounds and complex mixtures from natural origin, providing reliable results that indicate the method's feasibility.     

The frequency and geographic distribution of Giardia infections in ruminants in Switzerland

In various districts (Cantons) of Switzerland 815 calves (a few days up to 6 months old), 382 lambs and 20 young goats (both groups 1-6 months old) were selected randomly for a single coprological examination (flotation method) for Giardia infection. In average 26.6% of the calves, 29.8% of the lambs and 4 of 20 young goats excreted Giardia cysts. In 9 Cantons the percentages of cyst excretors among the calves varied between 15 and 32, but these differences were not significant. Further there were no significant differences in the frequency of cyst excretion between calves of 3 different breeds and 2 age groups (up to 3 months old and 3 to 6 months). The intensity of cyst excretion was high and varied in random samples between 4.1 x 10(3) and 3.0 x 10(5) cysts per g of faeces in calves and between 2.2 x 10(3) and 1.6 x 10(5) in lambs. In 5 of 7 farms where calves and lambs were maintained simultaneously both animal species were Giardia infected. As expected, trophozoites of Giardia isolates from calves and lambs belonged to the Giardia duodenalis type and could not be differentiated on a morphological basis. Giardia cysts from calves (average measurements: 13.7 x 9.1 microns) and lambs (13.8 x 9.2 microns) were indistinguishable both morphologically and morphometrically. The results indicate that Giardia infections are frequent and geographically widely distributed in calves and lambs in Switzerland.     

Reactivity of gluten detecting monoclonal antibodies to a gliadin reference material

People affected by coeliac disease need to adhere to a life-long gluten-free diet to avoid symptoms. ELISA-tests are seen as the mainstay for the detection of gluten in gluten-free food because of their sensitivity. They can, however, yield different gluten amounts depending on the antibody and reference material used. We compared the reactivity of three prominent mouse anti-gliadin-antibodies to a reference gliadin isolated from 28 common bred European wheat varieties. The reference material proteins were labelled with fluorescent dye Cy3. They were then separated by 2DE and transferred by Western blot onto low fluorescent PVDF-membranes, followed by incubation with the three primary anti-gliadin antibodies one by one. Detection of the reacting proteins used anti-mouse antibody which was labelled with fluorescent dye Cy5. The use of this technique made it possible to co-detect the 2DE-image of the reference material proteins (Cy3) and proteins reacting with the respective antibody (Cy5). The three investigated antibodies had dissimilar reactivities with different proteins of the reference gliadin. Antibodies R5 and PN3 reacted mainly with gliadin fractions, antibody 401.21 mainly with high molecular weight glutenins. The results confirm the individual specificity of these antibodies and demonstrate the importance of validating immunochemical methods for gluten detection.     

糖甜菜定域育种初步研究

试验结果表明，甜菜单粒种采种母根主、侧花枝种子的块根产量、含糖率不同。单粒种根头周边芽所形成的花枝种子比根头中央部芽的花枝种子所产生的块根含糖率高0．79-2．45度，表现趋势较一致；但主、侧花枝种子的块根产量高低表现趋势不一致。为提高含糖率，此方法可在甜菜单粒型品系育种中应用。     

Efficiencies of ryegrass and white clover herbage utilization in mixtures continuously grazed by sheep

A method for measuring the growth, senescence and defoliation fluxes in a mixture consisting of alternate rows of perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) was used to calculate the actual and potential efficiencies of grass and clover utilization under continuous sheep grazing. White clover contributed relatively more to the growth than to the herbage mass of the mixture and its relative growth rate was usually significantly greater than that of its companion grass. The primary reason for the greater potential efficiency of herbage use of the white clover component was, however, its lower rate of senescence. Greater potential efficiency was not reflected in a greater actual efficiency because the contribution of white clover to the total herbage removed was always smaller than its contribution to the growth flux of the mixture. Despite the commonly assumed sheep preference for white clover, the legume was usually the least defoliated species, presumably because of the lower surface height of white clover compared with grass, which resulted in a lower vertical availability of white clover, thereby restricting its defoliation rate. The positive net herbage accumulation observed with white clover at a constant sward surface height occurred partly through stolon and bud development and partly through the growth of undefoliated leaves from axillary growing points and contributed to a net herbage accumulation per unit dry weight in white clover greater than that in ryegrass. This resulted in an increase in the white clover content of the sward, which was observed independently by destructive measurements.     

Efficacy of Chondroprotective Food Supplements Based on Collagen Hydrolysate and Compounds Isolated from Marine Organisms

Osteoarthritis belongs to the most common joint diseases in humans and animals and shows increased incidence in older patients. The bioactivities of collagen hydrolysates, sulfated glucosamine and a special fatty acid enriched dog-food were tested in a dog patient study of 52 dogs as potential therapeutic treatment options in early osteoarthritis. Biophysical, biochemical, cell biological and molecular modeling methods support that these well-defined substances may act as effective nutraceuticals. Importantly, the applied collagen hydrolysates as well as sulfated glucosamine residues from marine organisms were strongly supported by both an animal model and molecular modeling of intermolecular interactions. Molecular modeling of predicted interaction dynamics was evaluated for the receptor proteins MMP-3 and ADAMTS-5. These proteins play a prominent role in the maintenance of cartilage health as well as innate and adapted immunity. Nutraceutical data were generated in a veterinary clinical study focusing on mobility and agility. Specifically, key clinical parameter (MMP-3 and TIMP-1) were obtained from blood probes of German shepherd dogs with early osteoarthritis symptoms fed with collagen hydrolysates. Collagen hydrolysate, a chondroprotective food supplement was examined by high resolution NMR experiments. Molecular modeling simulations were used to further characterize the interaction potency of collagen fragments and glucosamines with protein receptor structures. Potential beneficial effects of collagen hydrolysates, sulfated glycans (i.e., sulfated glucosamine from crabs and mussels) and lipids, especially, eicosapentaenoic acid (extracted from fish oil) on biochemical and physiological processes are discussed here in the context of human and veterinary medicine.     

Sensitivity of Penicillium digitatum and P. italicum to Postharvest Citrus Fungicides in California

ABSTRACT Penicillium digitatum isolates (326), collected in California citrus groves and packinghouses, were assayed qualitatively for their sensitivity to imazalil, thiabendazole, and o-phenylphenol. Eighteen typical triple-resistant isolates, acquired in each of 3 years (1988, 1990, and 1994), were assayed quantitatively for their sensitivity to each of the three fungicides. No significant differences were found in the mean sensitivity of the isolates collected in different years. However, the proportion of isolates that were resistant to all three fungicides increased from 43% in 1988 to 77% in 1990 and 74% in 1994. Imazalil-resistant biotypes of P. digitatum were isolated frequently in California packinghouses, while resistant P. italicum was rare. No fungicide-resistant biotypes of either species were collected from citrus groves. Wild-type P. italicum was slightly less sensitive than wild-type P. digitatum to all three fungicides. The concentration of imazalil producing 50% growth inhibition (EC(50)) was three times greater when the age of the P. digitatum assay inoculum was increased from 12 to 24 h. Activity of imazalil increased with pH of the assay medium in the range pH 5.1 to 5.9, reflecting the greater concentration of dissociated imazalil at the higher pH value.     

Evidence from the use of isozyme markers of competition in swards between short-leaved and long-leaved perennial ryegrass

Intraspecific competition in Lolium perenne was studied using a replacement series of a short-leaved and a long-leaved genotype that was managed under different cutting frequencies and nitrogen supplies. Leaf length, tiller density and yield were recorded 6 months after sowing. Since the two competitors were labelled at the PG1/2 locus, it was possible to identify unambiguously the origin of tillers in the mixtures and to estimate the average length of lamina of each genotype. The difference in leaf length between the two competitors was maintained under particular types of management and generated contrasting competitive abilities leading to evolution of the genotypic composition of the mixtures. Thus, under infrequent cutting, competition for light resulted in the domination of the long-leaved genotype and in a concomitant intragenotypic selection for leaf length. Supplying nitrogen fertilizer emphasized the phenotypic plasticity of the genotypes, particularly of the dominated one. Frequent cutting reduced competition for light and the change towards the short-leaved genotype was far less rapid. The yielding and competitive abilities of ryegrass genotypes and the implications for breeding are briefly discussed.