Short-term impact of dry olive mill residue addition to soil on the resident microbiota

The short-term response of the resident soil bacterial and fungal communities to the addition of 5% (w/w) of either dry olive mill residue (DOR), DOR treated with Phlebia sp. (PTDOR) or DOR previously extracted with water (WEDOR) was investigated. As opposed to bacteria, the diversity of fungi increased upon the amendments as assessed by denaturing gradient gel electrophoresis of 18S rDNA. Over the first 30 days, phospholipid fatty acids analyses indicated a gradual decrease in the relative abundances of Gram⁺ bacteria (from 44.8% to 37.9%) and a concomitant increase of Gram⁻ bacteria (from 37.3% to 51.2%) in DOR-amended soil. A considerable increase in the fungal/bacterial ratio was observed after 7 days in DOR, WEDOR and PTDOR-amended soils with respect to the control (0.316, 0.165 and 0.265, respectively, vs. 0.011). The overall microbial activity was stimulated by the amendments as indicated by the higher activity levels of both dehydrogenase and fluorescein diacetate hydrolase. These results indicate that DOR at the application level examined is not toxic on soil microorganisms.     

Solubilization of peroxidase (EC 1.11.1.7) from the rat intestine using cations

A rat intestine M-L fraction was exposed to several agents in order to obtain the solubilization of peroxidase. These included inorganic cations, detergents, hydrolytic enzymes and membrane depolarizing drugs. Inorganic cations and cationic detergents were effective in solubilizing the enzyme activity from the particulate fraction.     

Tracking the evolution of epialleles during neural differentiation and brain development: D-Aspartate oxidase as a model gene

We performed ultra-deep methylation analysis at single molecule level of the promoter region of developmentally regulated D-Aspartate oxidase (Ddo), as a model gene, during brain development and embryonic stem cell neural differentiation. Single molecule methylation analysis enabled us to establish the effective epiallele composition within mixed or pure brain cell populations. In this framework, an epiallele is defined as a specific combination of methylated CpG within Ddo locus and can represent the epigenetic haplotype revealing a cell-to-cell methylation heterogeneity. Using this approach, we found a high degree of polymorphism of methylated alleles (epipolymorphism) evolving in a remarkably conserved fashion during brain development. The different sets of epialleles mark stage, brain areas, and cell type and unravel the possible role of specific CpGs in favoring or inhibiting local methylation. Undifferentiated embryonic stem cells showed non-organized distribution of epialleles that apparently originated by stochastic methylation events on individual CpGs. Upon neural differentiation, despite detecting no changes in average methylation, we observed that the epiallele distribution was profoundly different, gradually shifting toward organized patterns specific to the glial or neuronal cell types. Our findings provide a deep view of gene methylation heterogeneity in brain cell populations promising to furnish innovative ways to unravel mechanisms underlying methylation patterns generation and alteration in brain diseases.     

Purification of intestinal peroxidase (EC 1.11.1.7) in the rat

Rat intestine peroxidase was solubilized from a M-L fraction with BaCl2 and purified by gel filtration. The peroxidase preparation obtained by this procedure was purified 150 folds as compared to the activity present in the crude homogenate.