Bound fatty acids modulate the sensitivity of bovine β-lactoglobulin to chemical and physical denaturation

Fatty acids are the natural ligands associated with the bovine milk lipocalin, β-lactoglobulin (BLG), and were identified by means of mass spectrometry. The naturally bound ligands were found to contribute to the stability of the proteins toward denaturation by both temperature and chaotropes. To assess the nature of the structural regions involved in this stabilization, the thermodynamic and kinetic aspects of the stability of various structural regions of the proteins were studied in the presence of bound palmitate, which is the most abundant natural ligand. Binding of a single palmitate molecule was found to affect not only the stability of the calyx region, where palmitate is bound, but also that of the region at the hydrophobic interface between the barrel itself and the long helix in the protein structure, where the thiol group of Cys121 is buried. This region is known to be essential for the stability of the BLG dimer and is relevant to the generation of "reactive monomers" that are involved in covalent and noncovalent polymerization of BLG and in the formation of covalent adducts with other milk proteins.     

Hydraulic characteristics of composted pig manure,perlite, and mixtures of them,and their impact on cucumber grown on bags

Composted pig manure (CPM), unmixed or in mixture with perlite, was evaluated as growing medium based on both physical characteristics and agronomic performance. The physical properties were determined in CPM, perlite (0.5–2.5 mm), and their 75:25, 50:50 and 25:75 mixtures by volume. It was found that unmixed CPM has a higher bulk density (BD) than perlite, a slightly lower air space at a suction of 10 cm, and a markedly higher content in easily available water (EAW), although the water content at a suction of 10 cm is the same in the two media. Mixing CPM with perlite by 50:50 seems to increase the actual water content and decrease the actual air content at container capacity, while the BD and the EAW score intermediate to those in the unmixed constituents. Simulation of the relative hydraulic conductivity (K_r) indicates a sharp decrease in the unsaturated hydraulic conductivity (K) of CPM, perlite and their mixtures as the media suction increases. The differences in the decrease rate of K_r between the tested substrates are rather small, with the most and least steep decrease being observed in perlite and the mixture by 50:50, respectively. The agronomic performance of CPM was assessed by growing greenhouse cucumber in bags filled with perlite up to 20 cm in height, CPM up to 10 or 20 cm in height, and their mixture by 50:50 up to the same heights as CPM. A media height of 10 cm in the bag resulted in appreciably higher yields than a height of 20 cm, regardless of the type of the substrate, while the latter had no significant effect on yield. This effect of media height was ascribed to the steep decrease in K, which was minimized in all media at higher suction levels than 10 cm, thereby severely restricting water availability in their upper layers (10–20 cm) when their height in the bag was 20 cm.     

Evaluation of Mobility and Dissipation of Mefenoxam and Pendimethalin by Application of CSTR Model and Field Experiments Using Bare and Tobacco Tilled Soil Columns

The soil mobility and dissipation of two pesticides with different physicochemical properties, namely mefenoxam, a systemic fungicide, and pendimethalin a selective herbicide, were determined in bare and tobacco tilled soil columns, which were installed in field conditions for over 125 days. Soil samples were collected at specific time intervals for a 125-day period and the rate of pesticide dissipation and leaching through the soil column was studied. The dissipation half-lives of mefenoxam from the top soil layer in tilled and bare soil columns were estimated at 10.3 and 13.1 days, respectively, while the corresponding half-lives for pendimethalin were 26.7 and 27.5 days, respectively. The dissipation of mefenoxam and pendimethalin from the top soil in tobacco cultivation was faster in comparison with bare soil; however, 120 days after their application, both pesticide residues were detected in the soil. Maximum concentrations of mefenoxam and pendimethalin were observed on the 15^th and 33^rd day, respectively, in the soil layer of 5–10 cm depth and on the 30^th day and 63^rd day, respectively, in the soil layer of 10–15 cm depth. Higher concentrations were observed in bare soil columns. The leaching of both pesticides was simulated with the continuous stirred tank reactor (CSTR) in series model. The simulated peak concentration and peak time for both pesticides fitted reasonably well to the experimental values.     

Genotype,ultraviolet irradiation,and harvesting time interaction effects on secondary metabolites of whole lettuce and browning of fresh-cut product

This study evaluated the importance of three factors (genotype, ultraviolet [UV] radiation and harvest time) and their interaction on phenolics, tannins, flavonoids and antioxidant activity of ‘Paris Island’ (romaine type) and ‘Sanguine’ (red leaf type) lettuce plants as well as on the severity of browning of their fresh-cut product. Ten genotypes had significant effect on secondary metabolite accumulation and higher levels were found in ‘Sanguine’, while no interaction effect was found among all the studied factors. Therefore, a new analysis of variance was performed for each genotype separately. The results showed that in ‘Paris Island’ the time of harvest had a significant effect on all measured parameters with higher levels for plants from the night harvest, while UV radiation had little effect on the plants’ biochemistry. On the contrary, in ‘Sanguine’, UV radiation significantly affected the concentration of secondary metabolites and antioxidant capacity, with higher levels for plants grown under UV-open film. Browning evaluation of fresh-cut product derived from the above plants showed that factors affecting secondary metabolite concentration in whole plants at harvest did not have a similar effect. The findings indicate that there are appropriate treatments, for each genotype, that could achieve high production of secondary metabolites and improve their antioxidant properties.     

Mediterranean diet and overall mortality differences in the European Union

OBJECTIVE: To assess whether the Mediterranean diet contributes to overall mortality differences and trends between Mediterranean and non-Mediterranean European Union (EU) countries.Design: Routinely recorded adjusted overall mortality and food availability data in Mediterranean and non-Mediterranean EU countries. A Mediterranean diet score designed a priori was used as instrument. SETTING: Fifteen EU countries in the 1960s and the 1990s. SUBJECTS: The general population in the 15 EU countries. RESULTS: The difference between Mediterranean and non-Mediterranean EU countries in a 7-point Mediterranean diet score was reduced from 2.9 in the 1960s to 1.6 in the 1990s. This reduction may underlie the reduction in the difference in general mortality between these countries, from about 100 deaths per 100,000 person-years in the early 1970s to about 50 deaths per 100,000 person-years in the 1990s. CONCLUSIONS: The decline in overall mortality in the 15 EU countries over the last 25 years is probably unrelated to diet. However, the gradual loss of the survival advantage of Mediterranean EU citizens, compared with other EU citizens, may be linked to the gradual abandonment by the former of their dietary traditions.     

Geographic distribution of chestnut feeding insects in Greece

Greece has been traditionally considered as a chestnut-producing country; yet, chestnut production has experienced a dramatic decline in the last decades, mainly due to chestnut blight Cryphonectria parasitica and chestnut feeding pests. Though chestnut blight is partially under control, little attention has been granted on chestnut feeding pests in Greece. Two species are described as major pests, i.e., Cydia splendana and Curculio elephas, without any information regarding their distribution. This study aimed to identify species diversity that feed on chestnuts trees using genetic markers and further to obtain insight into their distribution. Besides C. splendana and C. elephas, three additional species were determined namely Cydia fagiglandana, Curculio glandium as well as a Curculio sp. that has never been genetically determined before. The distribution of these species was inhomogeneous in Greece. These results demonstrate clearly the need for a carefully planned pest management approach adapted to the local mixture of chestnut feeding pests of each area rather than one general approach.     

The Paleozoic origin of enzymatic lignin decomposition reconstructed from 31 fungal genomes

Wood is a major pool of organic carbon that is highly resistant to decay, owing largely to the presence of lignin. The only organisms capable of substantial lignin decay are white rot fungi in the Agaricomycetes, which also contains non-lignin-degrading brown rot and ectomycorrhizal species. Comparative analyses of 31 fungal genomes (12 generated for this study) suggest that lignin-degrading peroxidases expanded in the lineage leading to the ancestor of the Agaricomycetes, which is reconstructed as a white rot species, and then contracted in parallel lineages leading to brown rot and mycorrhizal species. Molecular clock analyses suggest that the origin of lignin degradation might have coincided with the sharp decrease in the rate of organic carbon burial around the end of the Carboniferous period.     

A High Multi-drug Resistance to Chemically Unrelated Oomycete Fungicides in Phytophthora infestans

Mutants of Phytophthora infestans with high resistance to the amidocarbamates iprovalicarb and benthiavalicarb and to the cyanoimidazole cyazofamid were isolated after UV-mutagenesis and selection on media containing one of the above fungicides. In vitro fungitoxicity tests showed that all resistant strains presented a highly reduced sensitivity to both cyazofamid and to the amidocarbamates. Cross-resistance studies with other oomycete fungicides from different chemical groups showed that the mutation(s) for resistance to iprovalicarb (IPV), benthiavalicarb (BVC) and cyazofamid (CZF) also greatly reduced the sensitivity of mutant strains to the phenylamide metalaxyl, acetamide cymoxanil, morpholine dimethomorph, benzamide zoxamide and to chlorothalonil. A lower reduction of sensitivity of mutant strains to the strobilurins azoxystrobin, kresoxim-methyl, pyraclostrobin and trifloxystrobin, azolones famoxadone and fenamidone and to antimycin A was observed. A resistance correlation was not apparent for the dithiocarbamate propineb and phenylpyridinamine fluazinam. Studies of fitness parameters in the wild-type and mutant strains of P. infestans showed that most resistant isolates had significantly reduced sporulation and sporangial germination, but not in the differentiation of sporangia into zoospores. Pathogenicity tests on tomato seedlings showed that most resistant isolates were significantly less pathogenic compared to the wild-type parent strain. However, experiments on the stability of the resistant phenotypes did not show a reduction in resistance when the mutants were grown for more than eleven generations on inhibitor-free medium. This is believed to be the first report of high level multi-drug resistance in fungal pathogens to chemically unrelated fungicides inhibiting different sites of cellular pathway.