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.     

Antimicrobial potential of egg yolk ovoinhibitor, a multidomain Kazal-like inhibitor of chicken egg

Chicken egg ovoinhibitor is a multidomain Kazal-type serine protease inhibitor with unknown function. Comparison of expression between different tissues indicated that ovoinhibitor is highly expressed in the magnum and liver followed by the uterus, which secrete egg white, egg yolk, and eggshell precursors, respectively. The results also revealed that ovoinhibitor expression is increased in the liver during sexual maturation followed by a subsequent decrease in mature hens. Ovoinhibitor was purified from the egg yolk plasma from nonfertilized eggs using two consecutive affinity chromatographies and gel filtration. Purified egg yolk ovoinhibitor was shown to inhibit trypsin and subtilisin. It was shown that purified egg yolk ovoinhibitor exhibited antimicrobial activities against Bacillus thuringiensis . The results suggest that this anti-protease plays a significant role in antibacterial egg defense against Bacillus spp., preventing contamination of table eggs (nonfertilized eggs) and protecting the chick embryo (fertilized eggs).     

Effect of cis/trans isomerism of beta-carotene on the ratios of volatile compounds produced during oxidative degradation

beta-Carotene is, when cleaved, an important source of flavor and aroma compounds in fruits and flowers. Among these aroma compounds, the main degradation products are beta-ionone, 5,6-epoxy-beta-ionone, and dihydroactinidiolide (DHA), which are associated by flavorists and perfumers with fruity, floral, and woody notes. These three species can be produced by degradation of beta-carotene through an attack by enzyme-generated free radicals and a cleavage at the C9-C10 bond. This study investigated the influence of cis/trans isomerism at the C9-C10 bond on the production of beta-carotene degradation compounds, first with a predictive approach and then experimentally with different isomer mixtures. beta-Carotene solutions containing high ratios of 9-cis-isomers produced more DHA, suggesting a different pathway than for the transformation of all-trans-beta-carotene to ionone and DHA. These results are important in the search for financially viable processes to produce natural carotene-derived aroma compounds.     

Comparison of the chemical composition of extracts from Scutellaria lateriflora using accelerated solvent extraction and supercritical fluid extraction versus standard hot water or 70% ethanol extraction

The aqueous extract of American skullcap (Scutellaria lateriflora L. (S. lateriflora), Lamiaceae) has been traditionally used by North American Indians as a nerve tonic and for its sedative and diuretic properties. Recent reports stated that flavonoids and possibly amino acids are responsible for the anxiolytic activity. As a part of our search for environmentally friendly solvents to extract the active components from medicinal plants, we used S. lateriflora in a comparison of accelerated solvent extraction (ASE) using water, and supercritical fluid extraction (SFE) using CO2 and 10% EtOH as modifier, at different temperatures. Flavonoids and amino acids were quantified by HPLC-UV and HPLC-MS, respectively. The flavonoid content was compared with conventional extraction methods (hot water extraction and 70% ethanol). The use of ASE at 85 degrees C with water as solvent gave the best results for flavonoid glycosides and amino acids, whereas SFE gave higher yields of flavonoid aglycones. However, the results obtained for total flavonoids were not significatively superior to hot water extraction or 70% aqueous EtOH extract.     

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.     

Detailed physicochemical characterization of the 2S storage protein from rape (Brassica napus L.)

Chromatographic, chemical, and spectroscopic techniques were used to characterize the physicochemical properties of napin purified by preparative chromatography. The molar extinction coefficient was determined (epsilon = 0.56), and static and dynamic light scattering measurements enabled the average molecular weight (M(w) = 13919), the second virial coefficient (A(2) = 23.95 x 10(-)(5) mol cm(3) g(-)(2)), and the hydrodynamic radius (R(H) = 1.98 nm) to be determined. No conformational changes were observed by fluorescence and circular dichroism measurements in different buffers at pH 3, 4.6, 7, and 12, confirming the high pH stability of this protein. From MALDI-TOF analysis and after enzymatic digestion, it was found that this purified sample, extracted from the rapeseed variety Express, contained mainly isoform 2SS3_BRANA.     

Fatty acid distribution in muscle,liver, and gonads of rays (Dasyatis marmorata,Rhinobatos cemiculus,and Rhinoptera marginata) from the East Tropical Atlantic Ocean

If a great number of rays are fished in the Tropical East Atlantic Ocean for their caudal fins, only a small amount of ray flesh is processed. Among them, three species of rays, Dasyatis marmorata, Rhinobatos cemiculus, and Rhinoptera marginata, from the Mauritanian coast have been investigated for the fatty acid composition of their lipids. Gas chromatography and gas chromatography-mass spectrometry allowed identification of 50 molecules from muscles, livers, and gonads of these fishes. Principal component analysis, starting from >50 samples, reveals significant differences in various fatty acid distributions, related to the species and sex of the sampled fish. Some of them are preferentially present in one sex or in both species, whereas the occurrence of others characterizes the male and female of one or two species. The results show that rays are potential resources of polyunsaturated fatty acids (PUFA) and should be used in the diet of local populations. The lipidic fractions contained a high amount of PUFA (up to 30% of the total), mainly composed of docosa-4,7,10,13,16,19-hexaenoic acid, eicosa-5,8,11,14-tetraenoic acid, and eicosa-5,8,11,14,17-pentaenoic acid.     

Pharmaceutical and nutraceutical management of canine osteoarthritis: present and future perspectives

Osteoarthritis (OA) is one of the most common chronic musculoskeletal diseases and causes of lameness in the dogs. The osteoarthritic disease process involves the entire synovial joint, encompassing the synovium, cartilage and underlying bone. Joint failure results from an abnormal mechanical strain causing damage to normal tissue or failure of pathologically impaired articular cartilage and bone under the influence of normal physiological strain or a combination of both. In both cases, the end point is cartilage loss and joint impairment. Osteoarthritic chondrocytes show an altered phenotype characterised by an excess production of catabolic factors, including metalloproteinases and reactive oxygen species. These factors constitute potential therapeutic targets and some new drugs and nutraceuticals have been proposed to promote the return to homeostasis. Until now, the therapeutic management of OA in dogs has been dominated by nonsteroidal anti-inflammatory drugs, but some new compounds, including diacerhein, with potential structure-modifying effects, are already used to treat OA in humans and could be helpful to manage OA in the dog. In addition, novel nutraceuticals, such as avocado/soybean unsaponifiable substances, have shown symptomatic effects in knee OA in humans, and could offer an alternative to prevent OA progression. This paper provides an overview of recent discoveries in the pathophysiology and in the therapeutic management of osteoarthritis in dogs.     

Molecular epidemiology of white pine blister rust: recombination and spatial distribution

ABSTRACT Multilocus haplotypes (MLHs) were derived for the spermogonial (monokaryotic haploid) stage of Cronartium ribicola, the causal agent of white pine blister rust. Six random amplified polymorphic DNA loci and three single-strand conformational polymorphism markers were analyzed for 246 rust samples collected from two heavily infected white pine plantations. All cankers sampled were spatially located within the plantations. The hypothesis that spores are not locally disseminated was supported by the absence of any spatial clustering in the distribution of the MLHs. A large number of MLHs was found at both sites and the haplotypic diversity was close to the maximum (one) in both populations. All measures of recombination were not different from expectations under a scenario of sexual recombination. Genetic differentiation between the two sites was very low (theta = 0.023), yet it was significantly different from zero (P < 0.01). This analysis is in agreement with a scenario of extensive sexual recombination followed by some long-distance dispersal.     

Molecular characterization of <Emphasis Type="Italic">Pythium</Emphasis> group F isolates by ribosomal-and intermicrosatellite-DNA regions analysis

Pythium group F is a ubiquitous, though minor, pathogen in several soilless and soil cultures; investigations were carried out to analyze different regions of the DNA and better understand the nature of this group. Fourty-two isolates were obtained from a variety of plants (cucumber, lettuce, tomato) grown in soil or soilless cultures collected in various countries (Canada, Denmark, France, Norway, Sweden and United Kingdom). All Pythium group F isolates displayed amplified ITS1-5,8S-ITS2 ribosomal DNA region (rDNA) of similar length, whereas polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) revealed that, among the seven enzymes used, polymorphism was only identified with Hin6I. After cloning of ITS1-5,8S-ITS2 rDNA region from Pythium group F isolates that displayed restriction polymorphism patterns with Hin6I, comparisons of sequence and restriction mapping data showed a slight variation consisting in a single base change. Inter Simple Sequence Repeat (ISSR)-PCR method was also used to obtain data related to the entire genome and not only to a single DNA region. It identified repeated motifs in the genome of Pythium group F isolates. Two primers (CAC)₅ and (CCA)₅ detected polymorphism, and isolates were classified among 11 molecular clusters. The genetic diversity of this group was not correlated with the geographical locations or the host plants from which the isolates originated. Polymorphism of Pythium group F isolates pointed out by ISSR is discussed