γ-Aminobutyric Acid (GABA)-enriched Hemp Milk by Solid-state Co-fermentation and Germination Bioprocesses.

This study introduces the concept of developing a functional hemp drink enriched with γ-Aminobutyric acid (GABA) to enhance its nutritional value and functional properties utilizing Solid-State (SSF) co-Fermentation by Lactobacillus casei and Bacillus subtilis and germination bioprocesses. Bioprocesses may offer an alternative solution to challenges in hemp milk, such as product instability and the use of additives. Notably, the hemp milk produced through the germination for three days or co-fermentation processes yielded the highest GABA content of 79.84 and 102.45 mg/100 mL, respectively, compared to the untreated milk. These bioactive milk samples exhibited higher zeta potential and soluble protein content and also reduced solid particle sedimentation and droplet sizes (D4,3 and D3,2) compared to the untreated milk. Furthermore, the peptide, total phenolic content, and antioxidant activity of the produced GABA-enriched kinds of milk surpassed those of the untreated milk. Overall, the SSF and germination processes present a promising alternative for producing stable milk analogs with enhanced health-boosting properties.

Effect of Germination on Fatty Acid Composition in Cereal Grains.

Sprouted grains are gaining popularity as functional food ingredients. This study aimed to evaluate the lipid and fatty acid composition of eight sprouted grains (millet, amaranth, quinoa, wheat, rye, barley, buckwheat, and oat). The method used was germination for up to 72 h at temperatures ranging from 19-23 °C. In general, the lipid content increased in the various grains sprouted, providing a rich source of polyunsaturated fatty acids. The % oil yield ranged from 1.17 ± 0.02% in sprouted rye to 5.71 ± 0.26% in sprouted amaranth. Germinated oat showed the greatest increase in fat content, 54.3%, compared to the control. Polyunsaturated fatty acids were more prevalent in whole grains (46.9-75.6%) than saturated fatty acids (10.1-25.9%) and increased with sprouting. The primary fatty acids detected in the grains, in order of abundance, were linoleic, oleic, palmitic, linolenic, and stearic acids. Millet sprouts contained the lowest total saturated fatty acids and the highest polyunsaturated fatty acids. Amaranth had the highest amount of saturated fatty acids, while buckwheat contained the lowest quantity of polyunsaturated fatty acids. The lowest omega-6/omega-3 ratio was 7 to 1 in sprouted rye and 8 to 1 in sprouted barley.

Analysis of Fatty Acid Composition in Sprouted Grains.

A whole-grain diet is associated with the prevention of metabolic syndromes, including obesity, diabetes, and cardiovascular diseases. Sprouting improves the nutritional profile and bioactive properties of grains, which are important for use as raw ingredients in the food industry. The aim of this review was to examine the lipid and fatty acid composition of germinated grains. The methods discussed include germination and analytical procedures for determining fat and fatty acid contents of grains. The effects of sprouting on the fat content and storage stability of grains were also assessed. Lipid levels ranged from 1.43% to 6.66% in the sprouted grains. The individual fatty acid content of grains changed depending on the germination conditions (17-37 °C, 1-9 days). Limited findings showed that sprouting grains at higher temperatures (20-25 °C) and longer times generated a healthy balance of omega-6 and omega-3 fatty acids, which is beneficial to humans. Future studies are needed to determine the optimum incubation and germination periods specific to each grain to improve the omega-6/omega-3 ratio. Free fatty acids were produced more slowly and levels of oxidation products were lower in sprouted grains than in the raw ingredients when stored for a year. Additional studies are required to investigate the oxidative stability and shelf life of sprouted grains.

Matrix-Specific Effects on Caffeine and Chlorogenic Acid Complexation in a Novel Extract of Whole Coffea arabica Coffee Cherry by NMR Spectroscopy.

Coffee cherry is a rich source of caffeine and chlorogenic acids. In this study we investigate the structural analysis of caffeine-enriched whole coffee cherry extracts, CEWCCE by using 1H and 13C NMR spectroscopy. The changes in 1H chemical shift data in NMR spectra of CEWCCE compared to pure caffeine indicated the formation of complexes between caffeine and chlorogenic acids in aqueous solution. The effect of complexation on the peak position of caffeoylquinic acid and caffeine resonance with increasing addition of caffeine was investigated. 2D NOESY experiments show the presence of cross-peaks that are due to the proximity of chlorogenic acid and caffeine molecules in stable complexes in protic solvents. The quantification data of caffeine by 1H qNMR was found to be in close agreement with the data obtained by HPLC analysis.

Metabolite Profiling Analysis and the Correlation with Biological Activity of Betalain-Rich Portulaca grandiflora Hook. Extracts.

The aim of the study was to evaluate the possible correlation between the bioactivity and the phytochemical profile of four betalain-rich extracts from Portulaca grandiflora Hook. The HPLC-DAD-ESI-MS analysis indicated the presence of 19 betaxanthins and two betacyanins. The highest concentrations of betaxanthins (982 mg/100 g DE) and betacyanins (650 mg/100 g DE) were noticed in orange and purple flowers extracts, respectively. The HPLC-DAD-ESI-HRMS/MS analyses revealed the presence of a total of 71 compounds. Fifteen new betaxanthins and fifty other metabolites were identified for the first time. The antioxidant activity of the studied flower extracts increased in the sequence of yellow < orange < purple < red (0.066−0.176 mM TE/g DE). Betalains showed less effect on the antioxidant activity of extracts than other metabolites did. Extracts from yellow and orange flowers were more active against Gram-positive bacteria (MIC = 4−16 mg/L), whereas extracts from red and purple flowers were slightly more active against Gram-negative bacteria (MIC = 16−32 mg/L). All the extracts showed the same activity against yeasts (MIC = 32 mg/L). Betaxanthins were active against Gram-positive bacteria, whereas betacyanins were active against Gram-negative bacteria. Remaining metabolites also exhibited antimicrobial activities. The cytotoxicity assessment showed that the P. grandiflora extracts were non-toxic to normal VERO cells. No significant antiviral activity towards Human Herpesvirus type 1 was observed (62 µg/mL). Among the tested varieties, the purple one showed anticancer selectivity towards colon carcinoma cells (RKO).

Cranberry: Chemical Composition, Antioxidant Activity and Impact on Human Health: Overview.

Cranberries are a rich source of bioactive compounds that comprise a healthy diet. Cranberry is abundant in nutritional components and many bioactive compounds that have antioxidant properties. Both American (Vaccinium macrocarpon) and European (Vaccinium oxycoccus) cranberry species are rich in polyphenols such as phenolic acids, anthocyanins and flavonoids, and is one of the few fruits that is high in proanthocyanidins, which is linked to many health benefits. The review systematizes information on the chemical composition of cranberry, its antioxidant effect, and the beneficial impact on human health and disease prevention after cranberry consumption, and in particular, its effect against urinary tract inflammation with both adults and children, cardiovascular, oncology diseases, type 2 diabetes, metabolic syndrome, obesity, tooth decay and periodontitis, Helicobacter pylori bacteria in the stomach and other diseases. Additional research needs to study cranberry proteomics profiling, polyphenols interaction and synergism with other biologically active compounds from natural ingredients and what is important in formulation of new functional foods and supplements.

Dehydrogenation of Betacyanins in Heated Betalain-Rich Extracts of Red Beet (Beta vulgaris L.).

Betacyanins are a group of water-soluble red-violet compounds containing nitrogen in their structure. These are biosynthesized in red beetroot (Beta vulgaris L.), a widely consumed vegetable that contains significant amounts of nutritious and bioactive compounds which are also found in dietary supplements. This contribution presents results of betacyanin thermal oxidation (resulting in dehydrogenation) interrelated with decarboxylation in selected acetate/phosphate buffers at pH 3-8 and at 85 °C, which may be of particular significance for formulation and performance of foods. Most of the reaction products were detected at the highest concentrations in the acidic solutions (pH 3-4). The main dehydrogenation reaction pathways were monitored by LC-DAD-MS/MS and were associated with decarboxylation of the principal extract pigments, betanin/isobetanin and neobetanin, at carbon positions C-2 and C-17. Additional reactions are accompanied by the 2,15-decarboxylation processes at different dehydrogenation levels with 15-decarboxy-betanin and 2,15-bidecarboxy-betanin, structurally elucidated by NMR analysis, as the distinct indicators of this route type. For other novel pigments detected, 2,15-bidecarboxy-xanbetanin, 2,15-bidecarboxy-xanneobetanin and 2,15,17-tridecarboxy-neobetanin, additional high resolution mass spectrometric analyses were performed and confirmed their molecular formulas.

Quantification of Major Bioactive Constituents, Antioxidant Activity, and Enzyme Inhibitory Effects of Whole Coffee Cherries (Coffea arabica) and Their Extracts.

Coffee cherry is a rich source of chlorogenic acids (CGAs) and caffeine. In this study we examined the potential antioxidant activity and enzyme inhibitory effects of whole coffee cherries (WCC) and their two extracts on α-amylase, α-glucosidase and acetylcholinesterase (AChE) activities, which are targets for the control of diabetes and Alzheimer's diseases. Whole coffee cherry extract 40% (WCCE1) is rich in chlorogenic acid compounds, consisting of a minimum of 40% major isomers, namely 3-caffeoylquinic acids, 4-caffeoylquinic acids, 5-caffeoylquinic acids, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, 4-feruloylquinc acid, and 5-feruloylquinc acid. Whole coffee cherry extract 70% (WCCE2) is rich in caffeine, with a minimum of 70%. WCCE1 inhibited the activities of digestive enzymes α-amylase and α-glucosidase, and WCCE2 inhibited acetylcholinesterase activities with their IC50 values of 1.74, 2.42, and 0.09 mg/mL, respectively. Multiple antioxidant assays-including DPPH, ABTS, FRAP, ORAC, HORAC, NORAC, and SORAC-demonstrated that WCCE1 has strong antioxidant activity.

Extraction and Natural Bioactive Molecules Characterization in Spinach, Kale and Purslane: A Comparative Study.

Leafy green vegetables contain essential nutrients and are frequently consumed for their perceived health benefits. The purpose of this study was to profile the nutritional and natural bioactive phytochemical compounds extracted from freeze-dried spinach and kale and compare them with our previously published freeze-dried purslane results. Novel research suggests that these leafy greens contain an abundance of fatty acids, amino acids, organic acids, vitamins, and minerals, which can reduce the risk of many chronic diseases. LC-MS/MS screening identified 69 and 103 compounds in spinach and kale, respectively, including flavonoids, glucosinolates, and phenolic and organic acids. Out of a total of 26 flavonoids identified in the current study, only three were found in spinach. All three leafy greens showed nutritional and health benefits and the unique phytochemical compounds found only in purslane provide a strong basis to incorporate its distinct dietary benefits.

Identification of Phytochemical Compounds in Coffea arabica Whole Coffee Cherries and Their Extracts by LC-MS/MS.

The current work assessed the phytochemical contents of Arabica whole coffee cherry (WCC) and its two commercially available extracts: a minimum 40% chlorogenic acid (CGA; WCCE-1) and 70% caffeine (WCCE-2). Mass spectrometry analyses tentatively identified 219 phytochemicals in the three coffee samples, which is, so far, the largest number of identifications in a single study. A new group of CGA derivative namely caffeoylvaleroylquinic acid (CVQA) was identified in the three samples. Moreover, three 5-hydroxytryptamide derivatives (C20-5HT, C22-5HT, and C24-5HT) were identified in WCC but not in the extracts. Two groups of atractyligenin derivatives (carboxyatractyligenin and noncarboxyatractyligenin) were identified in the three samples. Furthermore, our results show that both extracts retained a large number of phenolic and other potentially bioactive compounds that were naturally present in whole coffee cherry (WCC).

Neurophysiological Effects of Whole Coffee Cherry Extract in Older Adults with Subjective Cognitive Impairment: A Randomized, Double-Blind, Placebo-Controlled, Cross-Over Pilot Study.

Bioactive plant-based compounds have shown promise as protective agents across multiple domains including improvements in neurological and psychological measures. Methodological challenges have limited our understanding of the neurophysiological changes associated with polyphenol-rich supplements such as whole coffee cherry extract (WCCE). In the current study, we (1) compared 100 mg of WCCE to a placebo using an acute, randomized, double-blind, within-subject, cross-over design, and we (2) conducted a phytochemical analysis of WCCE. The primary objective of the study was to determine the neurophysiological and behavioral changes that resulted from the acute administration of WCCE. We hypothesized that WCCE would increase brain-derived neurotrophic factor (BDNF) and glutamate levels while also increasing neurofunctional measures in cognitive brain regions. Furthermore, we expected there to be increased behavioral performance associated with WCCE, as measured by reaction time and accuracy. Participants underwent four neuroimaging scans (pre- and post-WCCE and placebo) to assess neurofunctional/metabolic outcomes using functional magnetic resonance imaging and magnetic resonance spectroscopy. The results suggest that polyphenol-rich WCCE is associated with decreased reaction time and may protect against cognitive errors on tasks of working memory and response inhibition. Behavioral findings were concomitant with neurofunctional changes in structures involved in decision-making and attention. Specifically, we found increased functional connectivity between the anterior cingulate and regions involved in sensory and decision-making networks. Additionally, we observed increased BDNF and an increased glutamate/gamma-aminobutyric acid (GABA) ratio following WCCE administration. These results suggest that WCCE is associated with acute neurophysiological changes supportive of faster reaction times and increased, sustained attention.

The Responses of Bioactive Betanin Pigment and Its Derivatives from a Red Beetroot (Beta vulgaris L.) Betalain-Rich Extract to Hypochlorous Acid.

Neutrophils produce hypochlorous acid (HOCl) as well as other reactive oxygen species as part of a natural innate immune response in the human body; however, excessive levels of HOCl can ultimately be detrimental to health. Recent reports suggest that betacyanin plant pigments can act as potent scavengers of inflammatory factors and are notably effective against HOCl. Comparison of the in vitro anti-hypochlorite activities of a novel betalain-rich red beetroot (Beta vulgaris L.) extract with its pure betalainic pigments revealed that the extract had the highest anti-hypochlorite activity, far exceeding the activity of all of the betalainic derivatives and selected reference antioxidants. This suggests that it may be an important food-based candidate for management of inflammatory conditions induced by excessive HOCl production. Among all pigments studied, betanidin exhibited the highest activity across the pH range.

Phytochemical composition and nutritional value of different plant parts in two cultivated and wild purslane (Portulaca oleracea L.) genotypes.

Purslane (Portulaca oleracea) is a weed naturally found in driveways, lawns, and fields and edible in many regions of Europe, Asia, the Middle East, Africa, and Australia. The purpose of this study was to compare the nutritional and phytochemical components of cultivated and wild purslane. Omega-3 contents of both purslane genotypes were comparable with 189.16 ± 25.52 mg/100 g dry weight and 188.48 ± 6.35 mg/100 g dry weight in cultivated and wild purslane leaves, respectively. Omega-6/omega-3 ratio (1:1-1:3) were low in both genotypes. However, high levels of oxalic acid were observed. Cultivated contained greater amounts of amino acids and vitamins than wild purslane. Of the 184 compounds identified in both genotypes by LC-MS/MS, including phenolic acids, organic acids, flavonoids, alkaloids, and betanin, more than 80 showed greater than two-fold abundance in the wild compared to cultivated purslane. Purslane has the potential to be cultivated as a food ingredient for nutraceutical applications.

Structural Study on Hypochlorous Acid-Mediated Chlorination of Betanin and Its Decarboxylated Derivatives from an Anti-Inflammatory Beta vulgaris L. Extract.

Hypochlorous acid (HOCl) produced by neutrophils is a part of the natural innate immune response system in the human body, but excessive levels of HOCl can ultimately be detrimental to health. Recent reports suggest that betacyanin plant pigments can act as potent scavengers of inflammatory factors and are notably effective against HOCl. In this contribution, chlorination mechanism and position of the electrophilic substitution in betacyanins was studied by high-resolution mass spectrometry and further structural analyses by NMR techniques, which completed the identification of the chlorinated betacyanins. For the study on the influence of the position of decarboxylation on the chlorination mechanism, a comparison of the chlorination position between betanin as well as 17-, and 2,17-decarboxylated betanins was performed. The structural study confirmed that the chlorination position in betanin occurs within the dihydropyridinic moiety at carbon C-18. Therefore, out of the aqueous free chlorine equilibrium species: HOCl, OCl-, Cl2, and Cl2O, the most potent chlorinating agents are HOCl and Cl2O postulated previously and the attack of the Cl⁺ ion on the carbon C-18 with a cyclic intermediate version is considered.

The Fructoborates: Part of a Family of Naturally Occurring Sugar-Borate Complexes-Biochemistry, Physiology, and Impact on Human Health: a Review.

Sugar-borates (SBs) are mono- or di-sugar-borate esters (SBEs) comprised of one or two monosaccharide molecules linked to a boron (B) atom. SBEs occur naturally in commonly consumed herbs, vegetables, fruits, seeds, and nuts and, other than greatly varying levels of B found in local drinking water, are the primary natural dietary sources of B-containing molecules in humans. To date, the most studied SBE is calcium fructoborate (CaFB). CaFB represents an important example of how organic B-containing molecules are significantly distinct from their inorganic counterparts. During these past two decades, CaFB has been researched for its physical and biochemical characteristics, safety, and clinical outcomes. Results of these researches are presented and discussed herein. CaFB has been characterized using Fourier-transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), high-performance thin-layer chromatography (HPTLC), nuclear magnetic resonance (NMR), liquid chromatography-multistage accurate mass spectrometry (LC-MSn), X-ray diffraction (XRD), Raman spectroscopy, and inductively coupled plasma (ICP) in non-biological and biological specimens. Potential health benefits of CaFB have been clinically investigated in pilot and efficacy studies demonstrating (i) significant reductions in knee discomfort and improved flexibility within 7, 14, and 90 days and (ii) significant effect on blood levels of inflammatory, cardiovascular, and other biomarkers. These studies support the use of CaFB as a dietary supplement for the management of joint discomfort. CaFB is presented here in order to illustrate how physiological benefits are imparted by distinct organic boron-containing molecules rather than solely by the element B itself. Considering recent National Health and Nutrition Examination Survey (NHANES) data reporting increases in age-related joint pain and an increasing elderly demographic, SBEs offer potential for safe, natural, and effective management of joint discomfort and improved mobility in human and animal health applications. Several of these studies may also open new opportunities for use of SBEs for health benefits beyond joint health.

Determination and Quantitation of Residual Solvents in Natural Food Ingredients Using a Static Headspace Gas Chromatography with Flame Ionization Detection and Mass Spectrometric Detection Method.

The determination of residual solvents is critical for quality control in food production and manufacturing processes. A gas chromatographic and mass spectrometric detection (MSD) method was developed and validated to complement the static headspace flame ionization detection (FID) technique for identification, confirmation, and quantitation of solvent residues in botanicals, using Coffeeberry extract and pomegranate powder as test samples. Relative standard deviations (RSDs) of less than 12% were obtained for residual solvents in water and dimethyl sulfoxide. Coffeeberry extract supplemented with10 µg/g of Residual Solvents Class 3 Mix showed MSD and FID recoveries mainly of 91 to 121% and 77 to 110%, respectively, while those samples supplemented with 100 µg/g of Residual Solvents Class 3 Mix displayed MSD and FID recoveries of 105 to 123% and 87 to 112%, respectively. For pomegranate samples supplemented with 10 µg/g of Residual Solvents Class 3 Mix, MSD and FID recoveries of 95 to 124% and 72 to 151% were observed, respectively. Those samples supplemented with 100 µg/g of Residual Solvents Class 3 Mix exhibited MSD and FID recoveries of 109 to 135% and 97 to 127%, respectively. Thus, the developed procedure was demonstrated to be suitable for the analysis of residual solvents in natural food ingredients and is expected to find numerous industrial applications (e.g., process control and impurity detection in final products).

Phytochemical and physical properties of blueberries, tart cherries, strawberries, and cranberries as affected by different drying methods.

The effects of hot-air drying (AD), freeze drying (FD) and Refractance Window drying (RWD) on the retention of anthocyanins, phenolics, flavonoids, vitamin C and B, and antioxidant capacity (ORAC) in blueberries, tart cherries, strawberries, and cranberries, as well as organic acids and proanthocyanins in cranberries and chlorogenic acid and catchins in blueberries were evaluated. Changes in color, glass transition, specific heat, and surface morphology of the dried fruits were also evaluated. The quality retention in the dried fruits was found to be product and drying method specific. The FD products exhibited a better retention of vitamin C, ORAC, anthocyanins, and phenolics than the AD and RWD fruits. The RWD produced samples with a less effective ORAC, anthocyanins, and phenolics retention, but a higher total vitamin B retention compared to the FD products. The AD-dried fruits displayed significantly lesser quality retention in most of the quality indexes measured in the study.

Effects of controlled germination on selected physicochemical and functional properties of whole-wheat flour and enhanced γ-aminobutyric acid accumulation by ultrasonication.

Using hard red spring (HRS), hard white (HW), and soft white (SW) wheat, this study examined how germination time affected the functionality of whole-wheat flour (WWF) and enhancement of γ-aminobutyric acid (GABA) content through ultrasonication. The falling number values significantly decreased and the glucose content increased by 227-357% after 15h of controlled germination. The setback value of WWF paste decreased from 654 to 6cP (HW), 690 to 9cP (SW), and 698 to 7cP (HRS), respectively, showing significant decreases of starch retrogradation in an aqueous system. The gluten quality and dough mixing performance of WWF after 5-15h of controlled germination was enhanced since gluten is less weakened during the dough heating stage of Mixolab testing. After a 72h germination, the GABA content increased by 339% of the non-sprouting counterpart. Furthermore, the GABA content in the ultrasound-treated SW sample was 30.7% higher than that without ultrasound treatment.

Enhancement of gama-aminobutyric acid (GABA) and other health-related metabolites in germinated red rice (Oryza sativa L.) by ultrasonication.

Red rice (Oryza sativa L.) that has a red (reddish brown) bran layer in de-hulled rice is known to contain rich biofunctional components. Germination is an effective technique to improve the nutritional quality, digestibility, and flavor of de-hulled rice. Ultrasonication, a form of physical stimulation, has been documented as a novel approach to improve the nutritional quality of plant-based food. This study was undertaken to test the use of ultrasound to enhance the nutritional value of red rice. Ultrasonication (5min, 16W/L) was applied to rice during soaking or after 66h germination. Changes of metabolites (amino acids, sugars, and organic acids) in red rice treated by ultrasonication were determined using a GC/MS plant primary metabolomics analysis platform. Differential expressed metabolites were identified through multivariate statistical analysis. Results showed that γ-aminobutyric acid (GABA) and riboflavin (vitamin B2) in red rice significantly increased after germination for 72h, and then experienced a further increase after treatment by ultrasound at different stages during germination. The metabolomics analysis showed that some plant metabolites, i.e. GABA, O-phosphoethanolamine, and glucose-6-phosphate were significantly increased after the ultrasonic treatment (VIP>1.5) in comparison with the untreated germinated rice. The findings of this study showed that controlled germination with ultrasonic stress is an effective method to enhance GABA and other health-promoted components in de-hulled rice.

Oxidative stress or redox signalling - new insights into the effects of a proprietary multifunctional botanical dietary supplement.

Recent interest has focused on maintenance of healthy levels of redox signalling and the related oxidants; these parameters are crucial for providing us with concrete nutritional targets that may help us to better understand and maintain "optimal health". Following the above hypothesis, we performed a pilot double-blind, crossover, placebo-controlled, single dose study to measure the dose-dependent effects of a proprietary plant-based dietary supplement labelled here as S7 (SPECTRA7), related to how it affected the cellular metabolic index (CMI) in healthy human participants (n = 8). We demonstrated using the electron spin resonance/electron paramagnetic resonance spectrometer NOXYSCAN that the administration S7 resulted in statistically significant, long-term, dose-dependent inhibition of mitochondrial and cellular reactive oxygen species generation by as much as 9.2 or 17.7% as well as 12.0 or 14.8% inhibition in extracellular nicotinamide-dinucleotide-phosphate oxidase system-dependent generation of O2•-, and 9.5 or 44.5% inhibition of extracellular H2O2 formation. This was reflected with dose-dependent 13.4 or 17.6% inhibition of tumour necrosis factor alpha induced cellular inflammatory resistance and also 1.7 or 2.3-times increases of bioavailable NO concentration. In this pilot study, we demonstrated the ability of a natural supplement to affect cellular redox signalling, which is considered by many researchers as oxidative stress. The design and activity of this proprietary plant-based material, in combination with the newly developed "CMI" test, demonstrates the potential of using dietary supplements to modulate redox signalling. This opens the door to future research into the use of S7 for modulation of inflammatory markers, for sports endurance or recovery applications.