Use of agricultural by-products in the development of an agro-energy chain: A case study from the Umbria region.

Use of agricultural and livestock by-products for anaerobic digestion (AD), in total or partial substitution of the maize silage was evaluated from an environmental and economical point of view. The evaluation process included three methodological interdependent and consequential steps: the chemical stage at laboratory and plant level, the environmental and economic steps developing the Life Cycle Assessment and Life Cycle Costing jointly. The laboratory test showed that the two mixtures prepared with by-products, in partial (MIX A) and total (MIX B) substitution of maize silage, did not show differences in bio-methane production compared to a reference mixture with the 33% of maize silage. All mixtures tested at full-scale plant, showed the same performances, resulting in a similar energy production. Environmentally, MIX B increased greenhouse gas credits derived from the avoided production of mineral fertiliser for the energetic crops, resulting also in better economic performances. The break-even transport distances follow the positive environmental pattern result, in contrast to what was found for the break-even transport distances from the economic point of view.

Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment.

Lineage commitment during embryonic stem cell (ESC) differentiation is controlled not only by a gamut of transcription factors but also by epigenetic events, mainly histone deacetylation and promoter DNA methylation. The DNA demethylation agent 5'-aza-2'-deoxycytidine (AzadC) has been widely described as an effective promoter of cardiomyogenic differentiation in various stem cell types. However, its toxicity and instability complicate its use. Therefore, the purpose of this study was to examine the effects of zebularine (1-(ß-D-ribofuranosyl)-1,2-dihydropyrimidin-2-1), a stable and non-toxic DNA cytosine methylation inhibitor, on mouse ESC (mESC) differentiation. Herein, we report that treating embryoid bodies, generated from mESCs, with 30 µM zebularine for 7 days led to greater cell differentiation and induced the expression of several cardiac-specific markers that were detected using reverse transcription-polymerase chain reaction (RT-PCR), real-time PCR, immunostaining and flow cytometry. Zebularine enhanced the expression of cardiac markers and the appearance of beating cells that responded to cardiac drugs, including ion channel blockers (diltiazem) and ß-adrenergic stimulators (isoproterenol). Gene promoter methylation status was assessed using methylation-specific PCR (MSP) and validated by bisulfite sequencing analysis. Global gene expression profiling using microarrays showed that zebularine-differentiated cells are distinct from control ESCs. Pathway analysis revealed an enhancement of cellular processes such as embryonic development, cardiovascular system development and function. In addition, the whole-cell proteins exhibited different profiles as analyzed by two-dimensional differential-in-gel-electrophoresis. Our results indicate that zebularine regulates mesodermal differentiation of mESCs, controls promoter methylation of crucial cardiac genes and may help to improve cardiomyogenic differentiation.