Cytokine concentrations^* in plasma and spleen of pigs fed T1 diet (sunflower meal) or T2 diet (camelina oil cakes).

<p>*Concentration of cytokines was measured by ELISA in samples of spleen and plasma collected at the end of the experiment, using R&D Systems kits (according to the manufacturer’s instructions). Results were expressed as picograms (pg) of cytokine/mg of spleen protein or/ml of plasma. Data are means ± SEM (n = 12).</p>a,b,c<p> = Means with different superscripts within a row are significantly different (P<0.0).</p><p>Cytokine concentrations^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0110186#nt110" target="_blank">*</a>} in plasma and spleen of pigs fed T1 diet (sunflower meal) or T2 diet (camelina oil cakes).</p

Phospho-p65 NF-κB expression in protein spleen lysate.

<p>The level of p65-NF-kB phosphorylation in spleen of pigs fed or not with camelina oil-cakes was determined by western blot and expressed as the ratio between phospho-p65 NF-κB and β-actin band intensities respectively. For each group of animals the mean values ± SEM were calculated and presented as histogram. Statistical analysis was performed using one-way ANOVA followed by Fisher test (*<i>P</i><0.05, T1 diet-control group (white column) versus T2 diet -Camelina group (grey column).</p

Effect of camelina oil-cakes on antioxidant enzymes expression.

<p>Spleen samples were taken at the end of the trial on day 33 and were analyzed for SOD, CAT and GPx mRNA expression by quantitative RT-PCR. Results are expressed as fold change after normalization of the expression of target gene to the mean of 2 internally reference genes expression. Values are the means ± SEM, from two replicates. Statistical analysis was performed using one-way ANOVA followed by Fisher test (*<i>P</i><0.05, T1 diet-control group (white column) versus T2 diet -Camelina group (grey column).</p

Effects of T1 diet (sunflower meal) or T2 diet (camelina oil cakes) on selected blood biochemical parameters^*.

<p>*Pigs received two different dietary fat treatments: T1 diet (12% sunflower meal) and T2 (12% camelina oil cakes) diet for 33d. At the end of the experiment plasma from 12 pigs/group was used to measure the blood biochemical parameters. Data are means ± standard error of the mean (SEM).</p>a,b<p> = Mean values within a row with unlike superscript letters were significantly different (P<0·05).</p><p>Effects of T1 diet (sunflower meal) or T2 diet (camelina oil cakes) on selected blood biochemical parameters^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0110186#nt106" target="_blank">*</a>}.</p

Effect of camelina oil-cakes on the antioxidant capacity in spleen.

<p>The antioxidant level in spleen samples derived from pigs fed with camelina oil-cakes or control was measured as antioxidant capacity (TAC) kit (QuantiChrom – BioAssay Systems, USA). Results are expressed as Trolox equivalent antioxidant capacity. Data are means ± SEM. ANOVA one-way test followed by Fisher test was performed to analyze the effect of the different treatments on TEAC level (*<i>P</i><0.05, T1 diet-control group (white column) versus T2 diet -Camelina group (gray column).</p

Effect of camelina oil-cakes on NO production in spleen.

<p>Synthesis of NO was determined by measuring the nitric oxide level in spleen of pigs fed or not with camelina oil-cakes using the Griess assay. Nitrite absorbance was measured at 550 using a microplate reader (Tecan Infinite 200Pro, Austria) and a NaNO₂ standard curve ranging from 0 to 100 µM. Concentrations were calculated based on the NaNO₂ range, expressed as µmole/L NO₂⁻. Values are the means ± SEM, from two replicates. Statistical analysis was performed using one-way ANOVA followed by Fisher test (*<i>P</i><0.05, T1 diet-control group (white column) versus T2 diet -Camelina group (grey column).</p

Fatty acid composition (g/100 g of fatty acids) of sunflower meal and camelina oil cakes.

<p>Fatty acid composition (g/100 g of fatty acids) of sunflower meal and camelina oil cakes.</p

Correlations between gene expressions in spleen.

<p>Pearson correlation coefficient procedure was used to establish relationships between gene expression of nuclear receptors PPARγ, NF-κB and signalling p38-MAPK, inflammation-related molecules and antioxidant defense enzymes in spleen of pigs received dietary camelina cakes. The red and green colour gradient from dark to light shows the degree of positive or negative correlations respectively in spleen of pigs treated or not with camelina diet.</p

Effect of camelina oil-cakes on signaling molecules expression in spleen.

<p>Spleen samples were taken at the end of the trial on day 33 and were analyzed for PPAR-γ, NF-κB, MAPK-p-38α and Nrf2 mRNA expression by quantitative RT-PCR. Results are expressed as change after normalization of the expression of target gene to the mean of 2 internally reference genes expression. Values are the means ± SEM, from two replicates. Statistical analysis was performed using one-way ANOVA followed by Fisher test (*<i>P</i><0.05, T1 diet-control group (white column) versus T2 diet -Camelina group (grey column).</p

Fatty acid composition of experimental diets (g/100 g of total fatty acids).

1<p>BW range 68.45 to 98.0 kg.</p><p>T1: diet containing 12% sunflower meal.</p><p>T2: experimental diet including 12% camelina oil-cakes.</p><p>Fatty acid composition of experimental diets (g/100 g of total fatty acids).</p