Effects of type of added salt and ionic strength on physicochemical and functional properties of casein isolates produced by electroacidification

A procedure developed for soybean protein precipitation which was based on electrodialysis was tested for the production of acid casein from reconstituted skim milk. In a previous paper, the performance of bipolar membrane electroacidification (BMEA) was evaluated under different conditions of ionic strength (micro(added) = 0, 0.25, 0.5, or 1.0 M) and added salt (CaCl(2), NaCl, or KCl) (1). The aim of this study, which is the complement of the work on evaluation of BMEA performance, was to evaluate the functionality of the protein isolates produced by BMEA and to compare the BMEA isolates to commercial isolates and an isolate produced by chemical acidification. It was not possible to show differences between the functional properties of isolates produced by BMEA, except at 1 M CaCl(2) micro(added), due to the variability of the isolates. However, the results showed that it is possible to obtain isolates similar to commercial isolates and that the addition of salt during the process does not induce variations in functional properties. From results on mineral concentrations, it appeared that the addition of monovalent cations did not influence the retention of monovalent or divalent cations in the BMEA isolates, while addition of divalent cations (CaCl(2)) influenced the retention of magnesium. According to previous results on evaluation of BMEA performances under different conditions of ionic strength and added salt, the difference observed for the BMEA isolate produced at 1.0 M CaCl(2) was confirmed.     

Adapting ecotoxicological tests based on earthworm behavior to assess the potential effectiveness of forest soil liming

Two laboratory experiments were carried out to determine the potential effects of lime application on the behavior of earthworms inoculated in an acidified forest soil from Vosges Mountains. Several field soils were studied: (i) a non-limed soil that had received decades of atmospheric acid depositions $({\text{pH}}_{{\text{H}}_2\text{O}}=3.9)$, (ii) an in situ limed soil that had been limed 6 years before at 2.5 t ha^?1 $({\text{pH}}_{{\text{H}}_2\text{O}}=4.2)$ and (iii) in vitro limed soils composed of non-limed soil mixed with several lime amounts in the laboratory corresponding to field rates of 1, 2.5, 5, 10 and 20 t ha^?1. Firstly, we adapted the earthworm avoidance test (ISO, 2006) by using Eisenia fetida as the model organism and Lumbricus castaneus, a local species, to determine earthworm preference between non-limed and limed soils. Secondly, a cast production (CP) study was performed according to Capowiez et al. (2009) with Lumbricus terrestris, in seven different treatments (non-limed soil, in situ limed and five in vitro limed soils with pH from 5 to 6.3). Both species avoided the non-limed soil in favour of in situ and in vitro limed soils for both species. Results of the CP bioassay showed that the bioturbation behavior of L. terrestris significantly increased with the increase of soil pH, following a dose–response curve. However, in the long term (>6 years), the application of lime was insufficient to significantly improve soil conditions for enhancing earthworm activity. In conclusion, we may recommend this kind of earthworm laboratory tests with field soils to assess any changes in soil quality over time due to liming application.     

Cross-sectional study on acrylamide hemoglobin adducts in subpopulations from the European Prospective Investigation into Cancer and Nutrition (EPIC) Study

Acrylamide exposure was investigated in subgroups of the EPIC study population (510 subjects from 9 European countries, randomly selected and stratified by age, gender, and smoking status) using hemoglobin adducts of acrylamide (HbAA) and its primary metabolite glycidamide (HbGA). Blood samples were analyzed for HbAA and HbGA by HPLC/MS/MS. Statistical models for HbAA and HbGA were developed including body mass index (BMI), educational level, and physical activity. A large variability in acrylamide exposure and metabolism between individuals and country groups was observed with HbAA and HbGA values ranging between 15-623 and 8-377 pmol/g of Hb, respectively. Both adducts differed significantly by country, sex, and smoking status. HbGA values were significantly lower in high alcohol consumers than in moderate consumers. With increasing BMI, HbGA in nonsmokers and HbAA in smokers decreased significantly. In the assessment of potential health effects related to acrylamide exposure, country of origin, BMI, alcohol consumption, sex, and smoking status should be considered.     

In vitro digestibility of α-casozepine, a benzodiazepine-like peptide from bovine casein, and biological activity of its main proteolytic fragment

α-Casozepine is a peptide, corresponding to the sequence 91-100 of the bovine α(s1)-casein, displaying anxiolytic activity in the rat. The α(s1)-casein tryptic hydrolysate containing this peptide decreases stress effects after oral administration in various species including man. Therefore, the stability of this peptide toward gastric and pancreatic proteases has been assessed by using pepsin, chymotrypsin/trypsin, Corolase PP, pepsin followed by chymotrypsin/trypsin or pepsin followed by Corolase PP. α-Casozepine was slowly degraded by chymotrypsin, much more sensitive to pepsin and Corolase PP but not completely destroyed after 4 h kinetics. The bonds in the region 91 to 95 of the α-casozepine were totally resistant to hydrolysis by all studied proteases. Surprisingly, a fragment, corresponding to the sequence 91-97 and found in all the hydrolysis media in significant amount, possessed an anxiolytic activity in three behavioral tests measuring this parameter. This peptide could participate in the in vivo activity of α-casozepine.     

Antioxidant properties of trans-epsilon-viniferin as compared to stilbene derivatives in aqueous and nonaqueous media

trans-Epsilon-viniferin, the dimer of resveratrol, extracted from Vitis vinifera, has been evaluated for its antioxidant capacity. Its properties have been compared to those of resveratrol and synthetic stilbenic derivatives (4-hydroxystilbene, 4,4'-dihydroxystilbene, 3,5-dihydroxystilbene, and trimethylresveratrol), in regard to their liposolubility using two media with different polarity. The bleaching of beta-carotene by lipoperoxyl (LOO.) radicals in an oil/water (O/W) emulsion and the scavenging of superoxide anions (O(-)(2) in dimethyl sulfoxide (DMSO) using 5,5-dimethyl-1-pyrroline-N-oxide as a spin trap were followed using UV-visible and electron paramagnetic resonance, respectively. Epsilon-viniferin exhibits the best antioxidant capacity in the DMSO/O(-)(2) polar system (IC(50) = 0.14 mM) while 4,4'-dihydroxystilbene presents the highest antioxidant capacity in the O/W/LOO. system (inhibition of beta-carotene bleaching, 82%). Partition coefficients and kinetics of partition between 1-octanol and water were measured to discuss the antioxidant efficiency of the compounds in relation with their chemical structure.     

Impact of casein gel microstructure on self-diffusion coefficient of molecular probes measured by 1H PFG-NMR

The translational dynamics of poly(ethylene glycol) (PEG) polymers with molecular weights (Mw) varying from 6x10(2) to 5x10(5) were investigated by pulsed field gradient NMR in casein suspensions and in gels induced by acidification, enzyme action, and a combination of both. For molecules with Mwor=8000, there was strong dependence of diffusion on PEG size and on the casein network structure as revealed by scanning electron microscopy images. The diffusion coefficients of the two largest PEGs were increased after coagulation by amounts that depended on the internal structure of the gel. In addition, the 527,000 g/mol PEG was found to deviate from Gaussian diffusion behavior to greater or lesser extents according to the casein concentration and the sample microstructure. The results are discussed in terms of network rearrangements.     

Species composition of arbuscular mycorrhizal fungi in two mountain meadows with differing management types and levels of plant biodiversity

Species composition of arbuscular mycorrhizal fungi (AMF) was analysed in two differently managed mountain grasslands in Thuringia (Germany). Arbuscular mycorrhizal fungi were studied in the roots of 18 dominant plant species from a total of 56 (32%). Additionally, spores of AMF were isolated from soil samples. Arbuscular mycorrhizal fungi species composition was analysed based on 96 sequences of the internal transcribed spacer of the nuclear ribosomal DNA, 72 originated from mycorrhizal roots, and 24 originated from AMF spores. Phylogenetic analyses revealed a total of 19 AMF species representing all genera of the Glomeromycota except Scutellospora and Pacispora. Despite a different farming intensity, resulting in remarkable differences concerning their plant species diversity (27 against 43 plant species), the diversity of AMF was found to be similar with 11 species on the intensively farmed meadow and ten species on the extensively farmed one. Nevertheless, species composition between both sites was clearly different. It thus seems likely that the AMF species composition, but not necessarily the species number, is related to above ground plant biodiversity in the system under study.     

Geographical distribution of Alzheimer's disease cases at birth and the geochemical profile of Saguenay-Lac-Saint-Jean/Québec,Canada (image project)

Accepting as founded the hypothesis of a multifactorial origin for the Alzheimer's disease (AD), we explore the possible links between the geographical distribution of AD cases according to their birth place and the geochemical profile of the Saguenay-Lac-Saint-Jean (SLSJ) territory in the province of Québec. The method used for factor analysis of correspondances (FAC) and another one based on the differences between the residential and municipal geochemical concentrations (DRMC) have been used for this purpose. The geochemical matrix of SLSJ relates the highest concentrations of nickel, copper and zinc to the sector surrounding the Jonquière municipality. Moreover, high concentrations of iron, lead and manganese were found in the northern sector of the Lac Saint-Jean. The DRMC method corroborates this strong correlation between iron, lead and manganese. Furthermore, these concentrations do not differ from the north-american geochemical standards. No single geochemical element seems to be associate with the spatial distribution of cases. It is, however, still possible that a certain synergistic effect between two or several elements could be implied in the development of the disease. According to the results of the soil analysis, there does not seem to be any association between the spatial distribution of cases and the concentration of aluminum, a chemical element that has been thought to be implied in the etiology of AD. These results are compared with those obtained by other workers in the micronesian islands, where abnormally high frequencies of amyotrophic lateral sclerosis and parkinsonism dementia have already been observed.     

Discrimination of recombinant and pituitary-derived bovine and porcine growth hormones by peptide mass mapping

Somatotropins, which are used in cattle for growth and lactating performances, are difficult to reliably detect because no direct method exists. Reversed-phase high-performance liquid chromatography (RP-HLC) coupled to electrospray ionization quadrupole mass spectrometry (ESI/MS) has been developed to separate and characterize the N-terminal peptides resulting from tryptic cleavage of natural and recombinant growth hormones from different species (bovine, porcine, and human) and suppliers. Conditions for tryptic digestion were optimized. This technique was found to be optimal to cleave efficiently the N-terminal peptide of the proteins without releasing too much noise from the matrix. Characterization of the peptides through ESI(+)-MS allowed natural and recombinant growth hormones from bovine and porcine species with N-terminal amino acid sequences differing from one amino acid residue to be discriminated. However, the studied human growth hormones had similar primary sequences that did not permit any discrimination between recombinant and natural forms, thus confirming the known identity of these hormones. Protein digestions with pepsin and chymotrypsin were also compared but were not conclusive due to the too small N-terminal peptides released after proteolysis.