BioTeSys
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Background: (6S)-5-Methyltetrahydrofolate ((6S)-5-MethylTHF) is the physiological folate form in biological fluids. Salts of (6S)-5-MethylTHF may have advantages compared to folic acid and are increasingly used in foods and supplements. Objective and design: The present study describes the physicochemical properties of the (6S)-5-MethylTHF-2Chol salt as a source of methylfolate with respect to solubility, conductivity, and melting point. The pharmacokinetics of (6S)-5-MethylTHF-2Chol and folic acid were compared in a randomized controlled double-blind cross-over study using a single equimolar oral dose of each of the folate substances. Results: The solubility of the dicholine salt was very high (650 mg/mL in H2O and 40 mg/mL in H2O under acidic conditions). The incremental area under the curve (iAUC0-8h) was significantly higher after the administration of (6S)-5-MethylTHF-2Chol compared to folic acid ([1.64-fold, P < 0.0001] for total folate and 2.56-fold higher for (6S)-5-MethylTHF [P < 0.0001]). Discussion and conclusions: The bioavailability of (6S)-5-MethylTHF-2Chol is higher compared to folic acid. The crystalline structure of (6S)-5-MethylTHF-2Chol and its water solubility are advantageous in terms of stability in nutraceutical products and absorption in the gut. (6S)-5-MethylTHF-2Chol is the source of folate that may enable the development of new applications.

Genomic instability markers are important hallmarks of aging, as previously evidenced within the European study of biomarkers of human aging, MARK-AGE; however, establishing the specific metabolic determinants of vascular aging is challenging. The objective of the present study was to evaluate the impact of the susceptibility to oxidation of serum LDL particles (LDLox) and the plasma metabolization products of nitric oxide (NOx) on relevant genomic instability markers. The analysis was performed on a MARK-AGE cohort of 1326 subjects (635 men and 691 women, 35-75 years old) randomly recruited from the general population. The Inverse Probability of Treatment Weighting causal inference algorithm was implemented in order to assess the potential causal relationship between the LDLox and NOx octile-based thresholds and three genomic instability markers measured in mononuclear leukocytes: the percentage of telomeres shorter than 3 kb, the initial DNA integrity, and the DNA damage after irradiation with 3.8 Gy. The results showed statistically significant telomere shortening for LDLox, while NOx yielded a significant impact on DNA integrity. Overall, the effect on the genomic instability markers was higher than for the confirmed vascular aging determinants, such as low HDL cholesterol levels, indicating a meaningful impact even for small changes in LDLox and NOx values.

Osteoarthritis (OA) is a prevalent degenerative joint disease, often affecting the elderly, characterized by cartilage degradation and bone remodelling, leading to pain, stiffness, and impaired movement. This 12-week, randomized, double-blind, placebo-controlled pilot study investigated the efficacy of a formulation containing collagen type II, glucosamine hydrochloride and chondroitin sulfate, in alleviating knee OA symptoms in subjects with mild to moderate knee pain. Fifty-four participants were enrolled, with 52 completing the study. Participants were assessed using the Knee injury and Osteoarthritis Outcome Score (KOOS), performance-based physical function tests and global assessment tests. Results indicated significant improvements in the verum group compared to placebo. KOOS subscales showed improvements in symptoms (8.16%, p < 0.05), sport/recreation (25.25%, p < 0.01), and quality of life (27.66%, p < 0.001). Stiffness improved by 14.68% (p < 0.05) in the verum group. Both groups exhibited significant pain reduction (KOOS pain, p < 0.001 and p < 0.01 for verum and placebo, respectively). Performance-based measures, such as the Chair Stand test, improved significantly in both groups, though other metrics showed no significant differences. The verum group exhibited a linear improvement over 12 weeks, contrasting with the placebo group's initial response and subsequent plateau. SF-36 Physical Component scores improved in both groups (p < 0.05), but mental health scores remained unchanged. Global assessment favoured the verum group, with more participants reporting OA improvement and recommending the product (p < 0.01). Biomarkers hsCRP and COMP showed no significant changes, likely due to sample variability. While the study did not achieve its primary endpoint, the nutritional formulation demonstrated an excellent safety profile and encouraging improvements in patient-reported symptoms and quality of life, warranting larger confirmatory trials.

Heartburn and acid regurgitation are main symptoms of gastro-esophageal reflux, a widespread complaint with a significant impact on quality of life (QoL). Refluthin® for Heartburn chewable tablets (Refluthin) are a symptomatic treatment option containing a combination of an antacid (CaCO3, MgCO3) and a polysaccharide-rich extract with mucoprotective substances from Opuntia ficus-indica cladodes. To investigate performance and safety of Refluthin in the rapid and long lasting relief of heartburn and acid related gastric discomfort under practical use conditions, a prospective, clinical, uncontrolled, open-label study was conducted. Adults with symptomatic heartburn, acid regurgitation, and/or recurrent acid related gastric discomfort took one tablet Refluthin up to four times/day as needed for up to 4 weeks. Endpoints were both time to onset and duration of symptom relief; reflux symptom intensity/frequency; global assessments (QoL, performance, satisfaction, usability, tolerability); and safety. 81/100 participants (81%) responded with a first symptom relief within ≤ 20 min in ≥ 50% of the individual applications. Long lasting effects of > 120 min were reported by 83/100 (83%) participants. Significant reductions in heartburn event frequency and intensity were seen within the 4 weeks of intermittent use (p < 0.0001, respectively). Global assessment results and safety-relevant findings were also favorable. Results thus demonstrated a distinct rapid and long lasting symptom relief after intake of Refluthin, with a safe and easy use. The significant reductions in frequency and intensity of heartburn events over time indicate sustained effects under treatment. These long-term effects might be explainable by soothing and protection of the irritated mucous membrane by Refluthin.

It is widely recognized that dietary polyphenols have a strong tendency to interact with proteins, specifically those rich in proline residues. So far, the impact of a matrix rich in polyphenols, like coffee, on the bioavailability of amino acids (AAs) and bioactive peptides (BAPs) from collagen hydrolysates (CH) has not been elucidated. This study aimed to compare the bioavailability of the signature AAs and BAPs from bovine CH dissolved in coffee in healthy human volunteers. The clinical trial was carried out as randomized, cross-over design. CH were dissolved in water or coffee, respectively, and provided as single dose in a fasting state. The uptake of the CH signature AAs hydroxyproline (Hyp), glycine (Gly) and proline (Pro) and BAPs (Pro-Hyp, Hyp-Gly, Gly-Pro-Hyp) into the bloodstream was followed over a period of 360 min. Plasma concentrations were determined with UPLC-MS/MS. The peak concentrations of the free signature AAs after intake of CH dissolved in water occurred between 60 and 120 min. Notably, there were significant variations in the increase of plasma concentrations over time of Hyp and Pro when CH was dissolved in coffee. However, when considering the uptake of BAPs, the pharmacokinetic endpoints iAUC, ΔCmax, and Tmax demonstrated similarity between water and coffee. This study emphasizes the importance of understanding how different food matrices, such as water vs. coffee, affect the absorption of bioactive compounds from dietary sources, in this case from CH. Ongoing research aims to explore the connection between food matrices, BAPs and human health with the goal of optimizing dietary interventions for promoting overall well-being.

Background Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) offer many health benefits, yet the absorption from standard ethyl ester (EE) formulation requires concurrent intake of a fatty meal. This study aimed to evaluate the absorption of EPA and DHA from a novel omega-3 formulation, Phospholipids + EPA/DHA (PL+), which combines high phospholipdi krill oil (KO) and EE, in comparison to a standard EE product under a low-fat dietary condition in healthy adults. Methods In this randomized, cross-over, single-dose study, the pharmacokinetic profiles of EPA and DHA from PL+ and EE product was assessed. Each products contained approximately 1200 mg EPA+DHA. Participants consumed a single dose of the capsules under a low-fat diet regimen, with plasma samples collected at baseline and over a 72-hour period after dosing. Following a 14-day washout period, participants crossed over to the alternate treatment. Plasma concentrations of EPA, DHA, and combined EPA+DHA were analyzed. Results Twelve participants (six women and six men) were included, completed all study visits and were included in the analyses. The participants' mean age was 34.3 years (SD = 12.4), and the mean BMI was 22.6 kg/m² (SD = 3.2). The baseline-corrected incremental area under the curve (iAUC0–12h) for combined EPA+DHA was significantly higher for PL+ (255.5 [SD = 81.4] μg/mL*h) compared to EE (34.2 [SD = 33.4] μg/mL*h; P < 0.001). The treatment ratio of iAUC0–12h for PL+ relative to EE was 10.5 (95 % CI: 6.1 – 18.1; P < 0.001). Similar patterns were observed for iAUC0–24h and iAUC0–72h for combined EPA+DHA, as well as for EPA and DHA individually. Sensitivity analyses by adjusting the minimal difference on dose between products yielded consistent findings. Conclusions The results indicate that PL+ technology significantly enhances the absorption of EPA and DHA compared to standard EE alone in a low-fat dietary condition. These outcomes suggest that the absorption of EE formulations can be significantly improved through combination with high phospholipid krill oil, as evidenced by the performance of the PL+ formulation.

Background: Omega-3 fatty acids, including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are polyunsaturated fatty acids (PUFAs) with notable health benefits. Due to limited physiological production and insufficient dietary supply, external supplementation is important. Objective: This study aimed to compare the pharmacokinetics and bioavailability of EPA and DHA in AvailOm® omega-3-lysine salt (Lys-FFA) versus standard ethyl ester (EE) and triglyceride (TG) formulations after a single oral dose in healthy subjects. Design: A randomized, three-way crossover study was conducted with 21 healthy subjects. Results: Twenty-one subjects (10 men, 11 women) completed the study. The average age was 41.7 years, and the mean body mass index was 23.0 kg/m2. The Lys-FFA formulation showed significantly higher uptake of omega-3 fatty acids (EPA+DHA combined and each individually) compared to EE. Specifically, Lys-FFA had a 9.33-fold (0-12 h) and 8.09-fold (0-24 h) higher bioavailability of EPA+DHA than EE and a 1.57-fold (0-12 h) and 1.44-fold (0-24 h) higher bioavailability than TG. ΔCmax and Tmax also favored Lys-FFA over EE. Discussion: Under fasting conditions, the absorption of EPA and DHA from EE is limited due to the need for enzymatic cleavage before absorption. This requirement is bypassed with Lys-FFA, which does not need cleavage. Conclusions: The study demonstrates that EPA and DHA lysine salt (Lys-FFA) offers superior bioavailability compared to EE and triglyceride forms, presenting a more effective supplementation option.

The implication of Torquetenovirus (TTV) in ischemic heart disease (IHD) has not been thoroughly explored. This study investigated the association between TTV viremia, pro-inflammatory cytokines, and IHD risk in an aging population. This cross-sectional study included 900 non-IHD subjects and 86 individuals with IHD (aged 55-75 years) selected from the MARK-AGE project. Results were verified in another independent Report-Age cohort, including 94 inpatients with chronic IHD and 111 inpatients with non-IHD (aged 65-96 years). Multivariable logistic regression in the MARK-AGE cohort revealed that male sex, TTV viremia ≥4log, Cu/Zn ratio, diabetes, hypertension, and smoking were significant IHD predictors. Notably, TTV viremia ≥4log independently increased the IHD risk (odds ratio [OR]: 2.51, 95% confidence interval [CI]: 1.42-4.43), confirmed in the Report-Age cohort (OR: 4.90, 95% CI: 2.32-10.39). In a RASIG subgroup, individuals with TTV viremia ≥4 log, both with and without IHD, exhibited increased plasma pro-inflammatory cytokine levels (IFN-γ, IL-1β, IL-6, IL-10, IL-12p70, TNF-α) compared to those with TTV viremia <4 log. No significant difference in cytokine production was observed between IHD patients and non-IHD with TTV viremia ≥4 log. A positive correlation between TTV viremia and DNA methylation estimator of leukocyte telomere length was observed in Report-Age patients. Additionally, IHD Report-Age patients with TTV viremia ≥4 log displayed higher NLR and SIRI index than those with TTV viremia <4 log. In conclusion, a high TTV viremia is associated with an elevated IHD risk in the older population, potentially arising from an augmented pro-inflammatory response and immunosenescence.

Scope: A mixture of (6S)-5-methyltetrahydrofolate-calcium salt ((6S)-5-MTHF-Ca) and folic acid (FA) from multimicronutrient supplements may show a dose-dependent effect on serum folate concentrations. Methods and results: The study compares fasting concentrations of serum folate spices after 8 weeks of either 400 or 800 µg day-1 of 1:1 folate mixture in 172 nonpregnant women. Serum (6S)-5-MTHF concentrations raise from a mean (SD) of 19.1 (13.4) to 73.9 (19.6) nmol L-1 in the 800 µg group and from 17.5 (9.4) to 54.5 (21.1) nmol L-1 in the 400 µg group (p < 0.001 within-group changes). The raise in serum (6S)-5-MTHF is stronger in the 800 µg compared to the 400 µg group (p < 0.001 between-group). The prevalence of FA concentrations ≥0.20 nmol L-1 increases between baseline and week 8 in both groups, but is not different between the groups (p = 0.116). The mean percentage of (6S)-5-MTHF of total serum folate increases in both intervention groups, but is not different between the groups at 8 weeks (95.5 (4.1)% versus 94.4 (5.7)%, p = 0.309). Conclusions: Supplementation of multimicronutrients with 800 µg folate mix for 8 weeks causes higher serum (6S)-5-MTHF concentrations, but not a higher prevalence of detectable FA compares to 400 µg folate.

The predictive value of the susceptibility to oxidation of LDL particles (LDLox) in cardiometabolic risk assessment is incompletely understood. The main objective of the current study was to assess its relationship with other relevant biomarkers and cardiometabolic risk factors from MARK-AGE data. A cross-sectional observational study was carried out on 1089 subjects (528 men and 561 women), aged 40-75 years old, randomly recruited age- and sex-stratified individuals from the general population. A correlation analysis exploring the relationships between LDLox and relevant biomarkers was undertaken, as well as the development and validation of several machine learning algorithms, for estimating the risk of the combined status of high blood pressure and obesity for the MARK-AGE subjects. The machine learning models yielded Area Under the Receiver Operating Characteristic Curve Score ranging 0.783-0.839 for the internal validation, while the external validation resulted in an Under the Receiver Operating Characteristic Curve Score between 0.648 and 0.787, with the variables based on LDLox reaching significant importance within the obtained predictions. The current study offers novel insights regarding the combined effects of LDL oxidation and other ageing markers on cardiometabolic risk. Future studies might be extended on larger patient cohorts, in order to obtain reproducible clinical assessment models.