“High in omega-3 fatty acids” Bologna type sausages stabilized with an aqueous-ethanol extract of Melissa officinalis.

A new formulation of bologna-type sausage enriched in ω-3 polyunsaturated fatty acids (PUFA) (8.75% linseed oil) was developed, using a lyophilized aqueous-ethanolic extract of Melissa officinalis. A comparison with the effectiveness of butylhydroxy anisole (BHA) synthetic antioxidant to decrease the oxidation of PUFAs was performed. The formulation increased the ω-3 PUFAs content, especially α-linolenic acid, decreasing significantly the ω-6/ω-3 ratio from 17.3 to 1.9, and also the Atherogenic Index and Thrombogenic Index (0.38–0.31 and 1.03–0.54, respectively). Modified sausages with BHA and Melissa extract showed significantly lower peroxides value (2.62 and 6.11 meqO2/kg) and thiobarbituric acid value (0.26 and 0.27 mg malondialdehyde/kg) and higher antioxidant capacity (hydrophilic fraction ABTS: 0.45 and 0.74 meq Trolox/g product; lipofilic fraction ABTS: 0.44 and 0.37 meq Trolox/g product) than those without these ingredients (16.49 meq O2/kg, 2.08 mg malondialdehyde /kg, 0.26 and 0.27 meq Trolox/g product, respectively). Sensorial tests showed that acceptability of the new formulations was similar to control products

Genome-wide microarray expression and genomic alterations by array-CGH analysis in neuroblastoma stem-like cells.

Neuroblastoma has a very diverse clinical behaviour: from spontaneous regression to a very aggressive malignant progression and resistance to chemotherapy. This heterogeneous clinical behaviour might be due to the existence of Cancer Stem Cells (CSC), a subpopulation within the tumor with stem-like cell properties: a significant proliferation capacity, a unique self-renewal capacity, and therefore, a higher ability to form new tumors. We enriched the CSC-like cell population content of two commercial neuroblastoma cell lines by the use of conditioned cell culture media for neurospheres, and compared genomic gains and losses and genome expression by array-CGH and microarray analysis, respectively (in CSC-like versus standard tumor cells culture). Despite the array-CGH did not show significant differences between standard and CSC-like in both analyzed cell lines, the microarray expression analysis highlighted some of the most relevant biological processes and molecular functions that might be responsible for the CSC-like phenotype. Some signalling pathways detected seem to be involved in self-renewal of normal tissues (Wnt, Notch, Hh and TGF-β) and contribute to CSC phenotype. We focused on the aberrant activation of TGF-β and Hh signalling pathways, confirming the inhibition of repressors of TGF-β pathway, as SMAD6 and SMAD7 by RT-qPCR. The analysis of the Sonic Hedgehog pathway showed overexpression of PTCH1, GLI1 and SMO. We found overexpression of CD133 and CD15 in SIMA neurospheres, confirming that this cell line was particularly enriched in stem-like cells. This work shows a cross-talk among different pathways in neuroblastoma and its importance in CSC-like cells

“High in omega-3 fatty acids” Bologna type sausages stabilized with an aqueous-ethanol extract of Melissa officinalis.

A new formulation of bologna-type sausage enriched in ω-3 polyunsaturated fatty acids (PUFA) (8.75% linseed oil) was developed, using a lyophilized aqueous-ethanolic extract of Melissa officinalis. A comparison with the effectiveness of butylhydroxy anisole (BHA) synthetic antioxidant to decrease the oxidation of PUFAs was performed. The formulation increased the ω-3 PUFAs content, especially α-linolenic acid, decreasing significantly the ω-6/ω-3 ratio from 17.3 to 1.9, and also the Atherogenic Index and Thrombogenic Index (0.38–0.31 and 1.03–0.54, respectively). Modified sausages with BHA and Melissa extract showed significantly lower peroxides value (2.62 and 6.11 meqO2/kg) and thiobarbituric acid value (0.26 and 0.27 mg malondialdehyde/kg) and higher antioxidant capacity (hydrophilic fraction ABTS: 0.45 and 0.74 meq Trolox/g product; lipofilic fraction ABTS: 0.44 and 0.37 meq Trolox/g product) than those without these ingredients (16.49 meq O2/kg, 2.08 mg malondialdehyde /kg, 0.26 and 0.27 meq Trolox/g product, respectively). Sensorial tests showed that acceptability of the new formulations was similar to control products

PANTHER classification by signalling pathway.

<p>The differentially expressed genes in both SK-N-DZ and SIMA CSC-like cells were classified by PANTHER and graphed. The percentage represents the number of genes altered against total number of genes involved in each pathway.</p

RT-qPCR for Sonic Hedgehog (A) and TGF-β pathways analysis (B) in SIMA CSC-like cells.

<p>The graphs represent the 2^−ΔΔCt values obtained by RT-qPCR for neurospheres. The dotted line indicates the 2^−ΔΔCt control (cell line) value equal to 1. Significance against control: p<0.05 (*); p<0.01 (**); p<0.001 (***) and p<0.0001 (****). Selected genes cover the most relevant components of each pathway. Results confirmed aberrant activation of Hh (A) and TGF-β (B) signalling pathways in CSC-like cells.</p

MicroRNA identified in array-CGH analysis.

<p>MicroRNA identified in array-CGH analysis.</p

Gene expression by RT-qPCR.

<p>The graphs represent the 2^−ΔΔCt values obtained by RT-qPCR for neurospheres. The dotted line indicates the 2^−ΔΔCt control (cell line) value equal to 1. Significance against control: p<0.05 (*); p<0.01 (**); p<0.001 (***) and p<0.0001 (****). <b>Corroboration of expression array (A).</b> Both SK-N-DZ (▪) and SIMA (▴) CSC-like cells showed a similar expression profile. <b>Stem Cell Markers expression (B).</b> Results confirmed a high overexpression of CSC markers in SIMA cell line after the neurosphere formation assay.</p

Upregulated (UR, red) and downregulated (DR, green) genes (>1 fold-change) in neurospheres compared to standard cells lines SK-N-DZ and SIMA (A), represented with MAplots, were the Y-axis represents the red/green intensity ratio “M” and the X-axis the average intensity “A”.

<p>Venn diagrams illustrating the number of differentially expressed genes in SK-N-DZ and SIMA neurospheres compared to standard cell lines (B). The overlapping area represents the set of genes altered in both cell lines.</p

Whole chromosome plots.

<p>Array-CGH from SK-N-DZ (A) and SIMA (B) cell lines. The X-axis represents the chromosomes while the Y-axis represent the normalize log2 ratio fluorescence intensity thresholds −1 (loss) and 1 (gain). The results show gains and losses of small chromosomal regions.</p