36 UPCYCLED INGREDIENTS
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Energising properties: Carbonylation levels of mitochondrial proteins Carbonylated mitochondrial proteins were definitively labelled with a specific fluorescent probe. Following the labelling process, these proteins were efficiently resolved on a 4-20% gradient SDS-PAGE. The proteins were then fixed to the gel, allowing for the clear detection of carbonylated proteins through fluorescence scanning at an emission wavelength of 647 nm. To visualise total proteins, the gel was post-
Figure 3: Quantification of Filaggrin levels. The levels of Filaggrin of each experimental group are expressed as relative values (% vs Control) and shown as mean +/- S.D. ***, p<0.001 – one-way ANOVA and Dunnett’s post-hoc test for multi-comparisons vs Stress group (alpha=0.05)
CONTROL STRESS (UV-A)
0.1% CHESTNUT BARK EXTRACT + STRESS
stained with SyproRuby™ (Life Technologies, USA), and high-quality digital images were captured by differential fluorescence at 595 nm. The staining of total proteins was consistently performed with the SyproRuby fluorescent reagent. The digital image acquisition for both carbonyl and total proteins was executed using the iBright system (Thermofisher Scientific). The signal from the carbonylated proteins was normalised to the total protein signal for each sample, resulting in a reliable Carbonyl Score.
Carbonyl Score (Sample X)
=
(Fluorescence Signal, Sample X) Prot. Totals
Carbonylated Prot. (Fluorescence Signal, Sample X)
0.03% WHITE TEA EXTRACT + STRESS
0.03% LIQUORICE LEAVES EXTRACT + STRESS
0.01% ALMOND SHELLS EXTRACT + STRESS
The Carbonyl Score was obtained for each sample by densitometric analysis of carbonylated proteins signal (fluorescence units) then normalised concerning the signal obtained with the total proteins and presented in Figure 1 as the average value and standard deviation from the mean per group. The Stress induced a significant increase in
0.08% GRAPEVINE WOOD EXTRACT + STRESS
carbonylation on mitochondrial proteins. The presence of the tested products significantly contrasted the stress-induced increase of carbonylation on mitochondrial proteins. Except for the grapevine extract, all the extracts show significant differences with the stressed condition.
IL-1A ■ Nuclear labelling (DAPI) ■
Figure 4: In situ visualisation of IL-1α levels by epifluorescence microscopy. The specific signal of IL-1α (red) labelling is visualised and superposed to the cellular nuclei (DAPI, in cyan)
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Hydration level: Filaggrin In situ detection of filaggrin levels was performed by epifluorescence microscopy, as shown in Figure 2. The Filaggrin signal is visualised in a colour range (low levels in dark, high levels in bright colours) and superposed to the cellular nuclei (DAPI, in cyan). As expected, the stress (UV-A), induced a decrease in Filaggrin levels. The presence of all extracts counteracted the stress-induced decrease in filaggrin levels. The filaggrin levels (%) are reported in a bar
graph representation as mean values +/- SD per experimental group, normalised to the control (Figure 3). The presence of all extracts significantly counteracted the stress-mediated decrease of filaggrin levels.
Figure 5: Quantification of IL-1α levels. The levels of IL-1α of each experimental group are expressed as relative values (% vs Control) and shown as mean +/- S.D. ***, p<0.001 – one-way ANOVA and Dunnett’s post-hoc test for multi-comparisons vs Stress group (alpha=0.05)
PERSONAL CARE September 2025
Anti-inflammatory properties: IL-1α In situ detection of IL-1α levels (visualised in red) was performed by epifluorescence microscopy, as seen in Figure 4. As expected, the stress (UV-A) increased the IL-1α levels. The presence of every extract counteracted the stress-induced decrease of IL-1a levels. Similar effects were observed with the reference 0.08% grapevine wood extract. The IL-1α levels (%) are reported in the bar graph representation as mean values +/- SD per
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IL-1a levels (RFU/surface, % of control)
Filaggrin levels (RFU/surface, % of control)
Control Stress (UV-A) 0.1% Chestnuts Bark Extract + Stress
0.03% White Tea Leaves Extract + Stress 0.03% Liquorice Leaves Extract + Stress 0.01% Almond Shells Extract + Stress 0.08% Grapevine Wood Extract + Stress
Control Stress (UV-A) 0.1% Chestnuts Bark Extract + Stress
0.03% White Tea Leaves Extract + Stress 0.03% Liquorice Leaves Extract + Stress 0.01% Almond Shells Extract + Stress 0.08% Grapevine Wood Extract + Stress
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