Nutrients 2023 , 15 , x FOR PEER REVIEW
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Nutrients 2023 , 15 , 3466
Figure 4. Paired associate learning (PAL): Mean (SEM) change from baseline to week 12 in total errors ( A ) and mean errors to success ( B ). Within-group di ff erences compared to baseline * p < 0.05 (Wilcoxon Signed-Rank Test) and between-group di ff erences * p < 0.05 (mixed two-way between– within groups ANOVA, outliers removed). 3.5. Blood Biomarkers Blood biomarkers were analyzed at baseline and after 12 weeks of intervention. The tryptophan metabolism was evaluated by measuring the tryptophan and kynurenine lev- els. No signi fi cant changes were observed for the LPHEAL9 group, while both tryptophan and kynurenine levels increased signi fi cantly from baseline to week 12 in the placebo group (Table 2). For tryptophan, the increase over time was signi fi cantly higher in the placebo group compared to the LPHEAL9 group. Di ff erent in fl ammation markers were also measured. The CRP level was below the detection limit (0.3 mg/L) for 10% of the sub- jects. Excluding these subjects, as well as subjects with increased CRP levels (>10 mg/L), indicating an ongoing infection/other non-healthy state, showed that the CRP level did not change over time. Non-signi fi cant changes over time were also observed for brain- derived neurotrophic factor and galectin-3 for both groups (Table S5). The pro-in fl amma- tory marker fractalkine increased in the placebo group ( p = 0.09), while a small decrease was observed in the LPHEAL9 group (54.3 pg/mL vs. − 0.78 pg/mL; p = 0.13 between groups). The anti-in fl ammatory marker TGF- β decreased signi fi cantly in the placebo group ( p = 0.03), but the level was unchanged in the LPHEAL9 group ( p = 0.176 between groups). Table 2. Kynurenine and tryptophan levels in plasma at baseline, week 12, and change over time in the ITT population (median, range). LPHEAL9 ( n = 64); placebo ( n = 62). Baseline 12 Weeks Change p -Value # Kynurenine (ng/mL) LPHEAL9 439.8 (210.3–1276) 450.8 (187.7–1295) − 3.9 ( − 603.3–413.4) Placebo 438.1 (176.7–1695) 497.5 (213.7–1173) 32.0 ( − 521.2–397.8) * 0.234 Tryptophan (µg/mL) LPHEAL9 9.80 (4.81–16.08) 10.11 (4.84–16.35) − 0.67 ( − 8.90–9.36) Placebo 9.90 (4.84–19.53) 11.53 (4.75–20.39) 2.31 ( − 11.92–12.06) *** 0.003 # Between-group di ff erences: independent samples from Mann–Whitney U Test. * p < 0.05; *** p < 0.001. Within-group: Wilcoxon Signed-Rank Test. 3.6. Intake of Study Product and Safety Compliance was overall good in the study and in the ITT population; the mean intake of the LPHEAL9 product was 98.5%, while the mean intake was 99.0% for the placebo product during the 12 week intervention period. Thirty-nine adverse events were reported during the study, 20 in the LPHEAL9 group and 19 in the placebo group. Two adverse events were possibly related to the intake Figure4. Paired associate learning (PAL): Mean (SEM) change from baseline to week 12 in total errors ( A ) and mean errors to success ( B ). Within-group differences compared to baseline * p < 0.05 (Wilcoxon Signed-Rank Test) and between-group differences * p < 0.05 (mixed two-way between–within groups ANOVA, outliers removed). 3.5. Blood Biomarkers Blood biomarkers were analyzed at baseline and after 12 weeks of intervention. The tryptophan metabolism was evaluated by measuring the tryptophan and kynurenine levels. No significant changes were observed for the LPHEAL9 group, while both tryptophan and kynurenine levels increased significantly from baseline to week 12 in the placebo group (Table 2). For tryptophan, the increase over time was significantly higher in the placebo group compared to the LPHEAL9 group. Different inflammation markers were also measured. The CRP level was below the detection limit (0.3 mg/L) for 10% of the subjects. Excluding these subjects, as well as subjects with increased CRP levels (>10 mg/L), indicating an ongoing infection/other non-healthy state, showed that the CRP level did not change over time. Non-significant changes over time were also observed for brain-derived neurotrophic factor and galectin-3 for both groups (Table S5). The pro-inflammatory marker fractalkine increased in the placebo group ( p = 0.09), while a small decrease was observed in the LPHEAL9 group (54.3 pg/mL vs. − 0.78 pg/mL; p = 0.13 between groups). The anti-inflammatory marker TGF- β decreased significantly in the placebo group ( p =0.03), but the level was unchanged in the LPHEAL9 group ( p = 0.176 between groups). Table2. Kynurenine and tryptophan levels in plasma at baseline, week 12, and change over time in the ITT population (median, range). LPHEAL9 ( n = 64); placebo ( n =62). Baseline 12Weeks Change p -Value # 439.8 (210.3–1276) 438.1 (176.7–1695) 450.8 (187.7–1295) 497.5 (213.7–1173) − 3.9 ( − 603.3–413.4) 32.0 ( − 521.2–397.8) * − 0.67 ( − 8.90–9.36) 2.31 ( − 11.92–12.06) *** 0.234 9.80 (4.81–16.08) 9.90 (4.84–19.53) 10.11 (4.84–16.35) 11.53 (4.75–20.39) 0.003 # Between-group differences: independent samples from Mann–Whitney U Test. * p <0.05; *** p < 0.001. Within- group: Wilcoxon Signed-Rank Test. 3.6. Intake of Study Product and Safety Compliance was overall good in the study and in the ITT population; the mean intake of the LPHEAL9 product was 98.5%, while the mean intake was 99.0% for the placebo product during the 12 week intervention period.
Kynurenine (ng/mL) LPHEAL9 Tryptophan ( µ g/mL) LPHEAL9 Placebo
Placebo
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