Association of TALS developmental disorder with defect in minor splicing component U4atac snRNA

The spliceosome, a ribonucleoprotein complex that includes proteins and small nuclear RNAs (snRNAs), catalyzes RNA splicing through intron excision and exon ligation to produce mature messenger RNAs, which, in turn serve as templates for protein translation. We identified four point mutations in the U4atac snRNA component of the minor spliceosome in patients with brain and bone malformations and unexplained postnatal death [microcephalic osteodysplastic primordial dwarfism type 1 (MOPD 1) or Taybi-Linder syndrome (TALS); Mendelian Inheritance in Man ID no. 210710]. Expression of a subgroup of genes, possibly linked to the disease phenotype, and minor intron splicing were affected in cell lines derived from TALS patients. Our findings demonstrate a crucial role of the minor spliceosome component U4atac snRNA in early human development and postnatal survival.     

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.     

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.     

Application of denaturing high-performance liquid chromatography (DHPLC) for the identification of fish: a new way to determine the composition of processed food containing multiple species

The identification of fish species in transformed food products is difficult because the existing methods are not adapted to heat-processed products containing more than one species. Using a common to all vertebrates region of the cytochrome b gene, we have developed a denaturing high-performance liquid chromatography (DHPLC) fingerprinting method, which allowed us to identify most of the species in commercial crab sticks. Whole fish and fillets were used for the creation of a library of referent DHPLC profiles. Crab sticks generated complex DHPLC profiles in which the number of contained fish species can be estimated by the number of major fluorescence peaks. The identity of some of the species was predicted by comparison of the peaks with the referent profiles, and others were identified after collection of the peak fractions, reamplification, and sequencing. DHPLC appears to be a quick and efficient method to analyze the species composition of complex heat-processed fish products.     

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.     

Differential transfer of organic micropollutants through intestinal barrier using Caco-2 cell line

Food seems to be one of the main ways of animal and human contamination with polycyclic aromatic hydrocarbons (PAHs) and dioxins. In vivo studies showed a blood absorption of these xenobiotics after their ingestion. Our work aimed at studying the in vitro transfer of PAHs and dioxins through intestinal barrier. Caco-2 cells were cultivated on permeable filters to measure transepithelial permeability of (14)C labeled 2,3,7,8-tetrachlorodibenzo-p-dioxin, benzo[a]pyrene, pyrene, and phenanthrene, which differed in their physicochemical properties. The results showed that the molecules were able to cross intestinal cell layers. All the molecules were detected associated with cells, even if the dioxin was the less uptaken compound. Phenanthrene appeared in basal media faster, and its level after a 6-h exposure was respectively 1.1, 2, and 7 times higher than pyrene, benzo[a]pyrene, and 2,3,7,8-tetrachlorodibenzo-p-dioxin levels. These findings suggest that intestinal epithelium plays a key role in selective permeability and then in bioavailibility of micropollutants.     

Conservation value of variable connectivity: aquatic invertebrate assemblages of channel and floodplain habitats of a central Australian arid-zone river, Cooper Creek

Rapidly expanding water resource development in arid and semi-arid zones of Australia threatens the flow regime and ecological integrity of the few large dryland rivers and their immense floodplains. Efforts to manage and conserve the surface waters of these rivers are hampered by limited scientific data on the ecology of their flora and fauna and on their responses to the high natural variability of flow regime that typifies dryland rivers. Irregular floods connect channel and floodplain wetlands to differing degrees and for varying periods of time but the ecological significance of this connectivity is poorly understood. On Cooper Creek, a large dryland river in central Australia, we explored the degree to which assemblage composition varied with connectivity and hydrological regime. Shortly after protracted regional flooding, we sampled aquatic macroinvertebrate assemblages from the principal microhabitats in 12 channel and floodplain wetlands. Ephemeral and temporary lakes tended to have fewer taxa than semi-permanent channel or terminal lake habitats. Although hydrological connection had only recently been lost for some wetlands, there was already evidence of divergence in aquatic macroinvertebrate assemblage composition. Disruption of the natural variability in connectivity and hydrological regime by excessive water abstraction or river-flow regulation threatens the ecological integrity and aquatic macroinvertebrate biodiversity of dryland rivers. Preservation of the irregular flow regime and sporadic connectivity underpins conservation of the mosaic of floodplain wetlands that play such a crucial role in the ecosystem functioning of rivers such as Cooper Creek.     

Influence of various phosphopeptides of caseins on iron absorption

The influence of the origin and kind of caseinophosphopeptide (CPP) on iron absorption was assessed by comparing a commercially available CPP mixture (CPPs) and derived chromatographic fractions with the purified, chemically phosphopeptide of beta-casein [beta-CN(1-25)] using a perfused rat duodenal loop system; gluconate iron was used as control. Only iron complexed to beta-CN(1-25) displayed a better bioavailability than gluconate iron. The results obtained with various chromatographic fractions indicated that phosphopeptides of different origins (alpha(s)- versus beta-caseins) display specific effects. These findings contribute to the explanation of the discrepancy about the role of caseinophosphopeptides on mineral bioavailability in vivo.     

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.     

Molecular modeling of syringyl and p-hydroxyphenyl beta-O-4 dimers. Comparative study of the computed and experimental conformational properties of lignin beta-O-4 model compounds

As a new approach for the study of the ultrastructure of lignin, the conformational preferences of lignin beta-O-4 model compounds have been investigated by molecular modeling. The computed results have been compared with the experimental data (X-ray crystal structures and (3)J(H)(alpha)(H)(beta) NMR coupling constant values) reported in the literature. This comparison has led to an improved understanding of the influence of the structure, stereochemistry, and intramolecular H-bonding upon the conformational properties of the beta-O-4 dimers. A large number of low-energy conformations have been predicted for the structures. It has also appeared that the conformational features are predominantly governed by local steric interactions rather than by differences in the H-bonding interactions. The threo and erythro forms differ significantly in their conformational features, with a preferential extended overall shape for the threo form in which the bulky aromatic groups are distant from each other.