Novel lines of Pax6-/- embryonic stem cells exhibit reduced neurogenic capacity without loss of viability

<p>Abstract</p> <p>Background</p> <p>Embryonic stem (ES) cells can differentiate into all cell types and have been used extensively to study factors affecting neuronal differentiation. ES cells containing mutations in known genes have the potential to provide useful in vitro models for the study of gene function during neuronal differentiation. Recently, mouse ES cell lines lacking the neurogenic transcription factor Pax6 were reported; neurons derived from these <it>Pax6</it>^-/-ES cells died rapidly after neuronal differentiation in vitro.</p> <p>Results</p> <p>Here we report the derivation of new lines of <it>Pax6</it>^-/-ES cells and the assessment of their ability to survive and differentiate both in vitro and in vivo. Neurons derived from our new <it>Pax6</it>^-/-lines were viable and continued to elaborate processes in culture under conditions that resulted in the death of neurons derived from previously reported <it>Pax6</it>^-/-ES cell lines. The new lines of <it>Pax6</it>^-/-ES cells showed reduced neurogenic potential, mimicking the effects of loss of Pax6 in vivo. We used our new lines to generate <it>Pax6</it>^-/-↔ <it>Pax6</it>^+/+chimeras in which the mutant cells survived and displayed the same phenotypes as <it>Pax6</it>^-/-cells in <it>Pax6</it>^-/-↔ <it>Pax6</it>^+/+chimeras made by embryo aggregation.</p> <p>Conclusions</p> <p>We suggest that loss of Pax6 from ES cells reduces their neurogenic capacity but does not necessarily result in the death of derived neurons. We offer these new lines as additional tools for those interested in the generation of chimeras and the analysis of in vitro ES cell models of Pax6 function during neuronal differentiation, embryonic and postnatal development.</p

Epilepsy Caused by an Abnormal Alternative Splicing with Dosage Effect of the SV2A Gene in a Chicken Model

Photosensitive reflex epilepsy is caused by the combination of an individual's enhanced sensitivity with relevant light stimuli, such as stroboscopic lights or video games. This is the most common reflex epilepsy in humans; it is characterized by the photoparoxysmal response, which is an abnormal electroencephalographic reaction, and seizures triggered by intermittent light stimulation. Here, by using genetic mapping, sequencing and functional analyses, we report that a mutation in the acceptor site of the second intron of SV2A (the gene encoding synaptic vesicle glycoprotein 2A) is causing photosensitive reflex epilepsy in a unique vertebrate model, the Fepi chicken strain, a spontaneous model where the neurological disorder is inherited as an autosomal recessive mutation. This mutation causes an aberrant splicing event and significantly reduces the level of SV2A mRNA in homozygous carriers. Levetiracetam, a second generation antiepileptic drug, is known to bind SV2A, and SV2A knock-out mice develop seizures soon after birth and usually die within three weeks. The Fepi chicken survives to adulthood and responds to levetiracetam, suggesting that the low-level expression of SV2A in these animals is sufficient to allow survival, but does not protect against seizures. Thus, the Fepi chicken model shows that the role of the SV2A pathway in the brain is conserved between birds and mammals, in spite of a large phylogenetic distance. The Fepi model appears particularly useful for further studies of physiopathology of reflex epilepsy, in comparison with induced models of epilepsy in rodents. Consequently, SV2A is a very attractive candidate gene for analysis in the context of both mono- and polygenic generalized epilepsies in humans

Integrating anaerobic digestion and pyrolysis for treating digestates derived from sewage sludge and fat wastes

P. 32603-32614The coupling of biological and thermal technologies allows for the complete conversion of wastes into energy and biochar eliminating the problem of sludge disposal. The valorisation of fatty residues as co-substrate in a mesophilic digester of a wastewater treatment plant was studied considering an integrated approach of co-digestion and pyrolysis. Four digested samples obtained from co-digestion of sewage sludge and butcher’s fat waste were studied by thermogravimetric analysis. The activation energy corresponding to the sludge pyrolysis was calculated by a non-isothermal kinetic. Arrhenius activation energy was lower for the pyrolysis of a digested grease sample (92 kJ mol−1 obtained by OFW and 86 kJ mol−1 obtained by Vyazovkin) than for the pyrolysis of sewage sludge and its blends (164–190 kJ mol−1 obtained by OFW and 162–190 kJ mol−1 obtained by Vyazovkin). The analysis of the integrated approach of anaerobic co-digestion and pyrolysis of digestates demonstrated that the addition of 3% (w/v) of fat to the feeding sludge results in a 25% increase in the electricity obtained from biogas (if a combined heat and power unit is considered for biogas valorisation) and increasing the fat content to 15% allows for covering all thermal needs for drying of digestate and more than doubles (2.4 times) the electricity production when the scenario of digestion and pyrolysis is contemplated.S

MULTILAYER HYDROREPELLENT SYSTEMS WITH CONTROLLED RELEASE OF ACTIVE AGENTS IN WATER AND SOIL

The invention relates to formulations for controlled release both in water and in soils, and to the components and processes that enable effective coating of cores in various forms. The technology used for implementing the invention, i.e., multilayer coating of cores with the addition of various additives and active ingredients, enables modulation of release of the active ingredients in the desired times and modes. Release is regulated according to the desired targets. The technology does not use any organic solvent, but is based upon a thermodynamic technology that uses the phase transitions through control of the heat introduced into or removed from the system. The invention uses low-cost compounds, mostly of natural derivation and in any case biodegradable. Hence, the materials and technology necessary for implementing the present invention have a low environmental impact

Automated image analysis and hyperspectral imagery with enhanced dark field microscopy applied to biochars produced at different temperatures

Biochar from agricultural biomasses and solid wastes represents a win-win solution for a rational waste management. Its sustainable usage requires the identification and standardization of biochar characteristics. The aim of this work was to identify the physical-chemical and spatial characteristics of biochars from pruning residues (PR), poultry litter (PL), and anaerobic cattle digestate (CD) at two pyrolysis temperatures (350 °C and 550 °C). The biochar characterization was carried out by applying emerging imaging techniques, 2D automated optical image analysis and hyperspectral enhanced dark-field microscopy (EDFM), and by SEM analysis. As predictable, the feedstock composition and the pyrolysis temperature strongly influence the physical structures of the biochar samples. Irrespective of charring temperature, PR biochar was mainly characterized by a broken and fragmented structure with an irregular and rough particle surface, completely different from the original PR wood cell. The EDFM imaging analysis evidenced the thermal degradation of PR vegetal products, composed primarily of hemicellulose, cellulose and lignin. On the contrary, small and regular particles with a smooth surface were produced by the PL pyrolysis, especially at 550 °C, due to the lower PL morphological homogeneity in comparison with the other biomasses. Finally, CD charring at both temperatures was characterized by changes in chemical composition, suggested by a lower pixel intensity. In conclusion, the emerging imaging techniques used in this study proved to be very effective in analyzing some properties of biochars, and can, therefore be considered as promising experimental strategies for detecting the feedstock and pyrolysis temperature of biochar

Identification de la mutation responsable de l'épilepsie réflexe photosensible chez le poulet

"Chantier qualité spécifique "Auteurs Externes" département de Génétique animale : uniquement liaison auteur au référentiel HR-Access "absen