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 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

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

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

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

Composition and calculated nutrient content of experimental diets (%).

1<p>BW range 68.45 to 98.0 kg.</p>2<p>mineral-vitamin premix (1%) supplied per kg diet as follows: vit. A 6000 IU, vit. D3 800 IU, vit. E 20 IU, vit. K1 1.0 mg, vit. B₁ 1.0 mg, vit. B₂ 3.0 mg, d-pantothenic acid 6.3 mg, niacin 10 mg, biotin 30 µg. vit. B₁₂ 20 µg, folic acid 0.3 mg, vit. B₆ 1.5 mg, Fe 80 mg, Zn 25 mg, Mn 30 mg, I 0.22 mg. Se 0.22 mg, Co 0.3 mg, antioxidants 60 mg. and maize starch as carrier.</p><p>Composition and calculated nutrient content of experimental diets (%).</p

Effect of camelina oil-cakes on inflammatory markers expression in spleen.

<p>Spleen samples were taken at the end of the trial on day 33 and were analyzed for COX-2, iNOS and eNOS 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