Differing Effects of Mechanical Dough Development and Sheeting Development Methods on Aggregated Glutenin Proteins

Dough development using sheeting and mechanical dough development (MDD) were compared with respect to the effect the mixing method had on the molecular size distribution and degree of protein thiol exposure of the aggregated glutenin proteins. Although sheeting imparts a lower rate of work input on doughs than does MDD mixing, changes in protein aggregation patterns during mixing were similar for both methods of dough development, indicating that protein disaggregation was important in the process of dough development. In both systems, a reduced rate of change in the protein aggregation patterns was associated with optimum dough development. The MDD mixing was characterized by increasing exposure of the thiol groups on the SDS‐insoluble glutenin during mixing while the sheeting process resulted in fewer exposed thiol groups on both SDS‐soluble and SDS‐insoluble glutenin proteins. This suggested that disulfide bond rupture may not be a required process in dough development and that high effective stresses per se may not be required to develop doughs. This is consistent with a model for dough development that does not require extensive covalent bond rupture but instead involves mainly rupture and reformation of noncovalent interactions such as hydrophobic bonds and hydrogen bonds between protein chains.     

Effect of host resistance on genetic structure of core and accessory chromosomes in Irish <Emphasis Type="Italic">Zymoseptoria tritici</Emphasis> populations

In agricultural pathosystems resistant cultivars are typically only temporarily effective, as widespread growth of said cultivars drives selection for pathogen genotypes capable of infecting them. A gene-for-gene interaction between Z. tritici and wheat has been demonstrated for one cultivar; however results of studies into the relevance of these interactions in the field remain inconsistent. Because genetic drift does not appear to occur between Z. tritici populations that are not widely geographically separated, according to neutral genetic theory if adaptation to different host cultivars is occurring, reduced genetic variation, and some differentiation between populations sourced from different cultivars should be observed. Selectively neutral microsatellite markers were used to genotype 260 isolates of Z. tritici taken from two naturally infected randomized block trials of four different cultivars, representing a spectrum of resistance to Z. tritici from susceptible to resistant. By calculating genetic parameters such as overall heterozygosity and F _ST from this genotypic data, the presented study aimed to determine if genetic drift or host selection is impacting on the genetic structure of the Irish Z. tritici population. Results indicated that diversity was distributed almost entirely within, rather than among populations, with little or no differentiation, and almost no clone isolates were present in the dataset. However this result was not reflected in the accessory chromosomes, where evidence of minor but significant genetic structure was found. This lack of structure in the core chromosomes and weak structure in the accessory chromosomes confirms that forces of genetic drift and selection are minor compared to sexual reproduction, in concurrence with multiple previous studies on other populations worldwide.     

Identification of a Pro-Angiogenic Potential and Cellular Uptake Mechanism of a LMW Highly Sulfated Fraction of Fucoidan from Ascophyllum nodosum

Herein we investigate the structure/function relationships of fucoidans from Ascophyllum nodosum to analyze their pro-angiogenic effect and cellular uptake in native and glycosaminoglycan-free (GAG-free) human endothelial cells (HUVECs). Fucoidans are marine sulfated polysaccharides, which act as glycosaminoglycans mimetics. We hypothesized that the size and sulfation rate of fucoidans influence their ability to induce pro-angiogenic processes independently of GAGs. We collected two fractions of fucoidans, Low and Medium Molecular Weight Fucoidan (LMWF and MMWF, respectively) by size exclusion chromatography and characterized their composition (sulfate, fucose and uronic acid) by colorimetric measurement and Raman and FT-IR spectroscopy. The high affinities of fractionated fucoidans to heparin binding proteins were confirmed by Surface Plasmon Resonance. We evidenced that LMWF has a higher pro-angiogenic (2D-angiogenesis on Matrigel) and pro-migratory (Boyden chamber) potential on HUVECs, compared to MMWF. Interestingly, in a GAG-free HUVECs model, LMWF kept a pro-angiogenic potential. Finally, to evaluate the association of LMWF-induced biological effects and its cellular uptake, we analyzed by confocal microscopy the GAGs involvement in the internalization of a fluorescent LMWF. The fluorescent LMWF was mainly internalized through HUVEC clathrin-dependent endocytosis in which GAGs were partially involved. In conclusion, a better characterization of the relationships between the fucoidan structure and its pro-angiogenic potential in GAG-free endothelial cells was required to identify an adapted fucoidan to enhance vascular repair in ischemia.     

Heparanase and Syndecan-4 Are Involved in Low Molecular Weight Fucoidan-Induced Angiogenesis

Induction of angiogenesis is a potential treatment for chronic ischemia. Low molecular weight fucoidan (LMWF), the sulfated polysaccharide from brown seaweeds, has been shown to promote revascularization in a rat limb ischemia, increasing angiogenesis in vivo. We investigated the potential role of two heparan sulfate (HS) metabolism enzymes, exostosin-2 (EXT2) and heparanase (HPSE), and of two HS-membrane proteoglycans, syndecan-1 and -4 (SDC-1 and SDC-4), in LMWF induced angiogenesis. Our results showed that LMWF increases human vascular endothelial cell (HUVEC) migration and angiogenesis in vitro. We report that the expression and activity of the HS-degrading HPSE was increased after LMWF treatment. The phenotypic tests of LMWF-treated and EXT2- or HPSE-siRNA-transfected cells indicated that EXT2 or HPSE expression significantly affect the proangiogenic potential of LMWF. In addition, LMWF increased SDC-1, but decreased SDC-4 expressions. The effect of LMWF depends on SDC-4 expression. Silencing EXT2 or HPSE leads to an increased expression of SDC-4, providing the evidence that EXT2 and HPSE regulate the SDC-4 expression. Altogether, these data indicate that EXT2, HPSE, and SDC-4 are involved in the proangiogenic effects of LMWF, suggesting that the HS metabolism changes linked to LMWF-induced angiogenesis offer the opportunity for new therapeutic strategies of ischemic diseases.     

Weak effects of habitat type on susceptibility to invasive freshwater species: an Italian case study

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Nomenclature and genetic relationships of apples and pears from Terceira Island

Heritage apple (Malus domestica Borkh. hybrids) and pear (Pyrus communis L. hybrid) trees grow in villages throughout Terceira Island, Azores, Portugal. Some of these pears have different names but similar morphology. The objective of this study was to determine synonymy, homology, and phylogeny of apples and pears from Terceira and to examine potential relationships of the island pears with standard apples and pears of Portuguese or American descent. Nine apple microsatellite markers were used to determine genetic relationships. Distance- and parsimony-based cluster analysis grouped these genotypes into separate apple and pear clades. The Terceira apples were divided into two clades: the maçā and the reineta-reinette. Among the 17 heritage apple genotypes, seven unique accessions were identified and four groups of synonyms, or possibly clones, were detected including: ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares; ‘Reineta Castanha’ and ‘Reineta Verde Miuda’; ‘Maçā Pêra,’ ‘Maçā Calhau’, ‘Pêro Branco’ from Salga and from Terra-Chā and ‘Maçā Marmelo’; and the five genotypes ‘Maçā Sao Joao’, ‘Malápio Rosa’, ‘Maçā Gaspar’, ‘Maçā Branca’ and ‘Maçā Pato’. In addition, two homonyms were detected. ‘Pêro Vermelho’ from Terra Chā was a separate genotype from a tree from Doze Ribeiras of the same name, but Pêro Branco from Terra Chā appears to be a clone that can be distinguished by an additional allele at CH1F07a from a tree with that name from Salga. One pair of apple clones, ‘Reineta Agosto’ and ‘Reineta Verde’ from Altares appear to be derived from an unreduced gamete of ‘Golden Delicious.’ Another apple genotype ‘Maçā Acida’ could be a sibling of the ‘Maçā Pêra’ clonal group. Other tested standard apples from the US genebank were unrelated to Terceira genotypes. Of the seven heritage pears, five unique genotypes and one pair of synonyms were detected. ‘Pêra Papo Pintassilgo’ from Raminho and ‘Pêra Vermelha’ from the nursery of Serviço de Desenvolvimento Agario da Terceira (SDAT) were synonyms. ‘Passans du Portugal’ was related to ‘Pêra Cabaca’ but other standard pears from the US genebank were unrelated to Terceira genotypes. Future studies will include additional apple and pear cultivars from other Islands of the Azores and continental Portugal, and wild Asian species to further explore genetic relationships.     

Do meadow birds profit from agri-environment schemes in Dutch agricultural landscapes?

Since 1992 the European Union helps member states to reverse the loss of biodiversity in agricultural landscapes by the financial support of agri-environment schemes. Long-term studies investigating the effects of these schemes are an essential prerequisite for the development of an effective policy to restore biodiversity on farmland. In Dutch meadow landscapes almost all agri-environment schemes focus on the restoration of meadow bird populations by postponement of the mowing date. Between 1990 and 2002 we measured long-term changes in meadow bird densities in areas with and without agri-environment schemes in the Netherlands, both before and after the start of the contract. During these years bird territories were surveyed during five field visits between 15 March and 15 June. Densities of black-tailed godwit (Limosa limosa), and redshank (Tringa totanus) were higher in the areas with management agreements, but these differences were already present before the start of the contracts. After the start of the management contracts densities of black-tailed godwit and oystercatcher (Haematopus ostralegus) did not increase, while those of lapwing (Vanellus vanellus) and redshank even declined relative to the control areas. It is concluded that the current agri-environment schemes are not sufficient to restore meadow bird populations in Dutch agricultural landscapes. In addition to the prescribed postponement of the mowing date, it is probably necessary to raise groundwater levels and to reduce fertilization to allow for the development of an open vegetation structure that will increase chick survival to sufficiently high levels.