Sportomics method to assess acute phase proteins in Olympic level athletes using dried blood spots and multiplex assay.

Sportomics is a subject-centered holistic method similar to metabolomics focusing on sports as the metabolic challenge. Dried blood spot is emerging as a technique due to its simplicity and reproducibility. In addition, mass spectrometry and integrative computational biology enhance our ability to understand exercise-induced modifications. We studied inflammatory blood proteins (Alpha-1-acid glycoprotein-A1AG1; Albumin; Cystatin C; C-reactive protein-CRP; Hemoglobin-HBA; Haptoglobin-HPT; Insulin-like growth factor 1; Lipopolysaccharide binding protein-LBP; Mannose-binding lectin-MBL2; Myeloperoxidase-PERM and Serum amyloid A1-SAA1), in 687 samples from 97 World-class and Olympic athletes across 16 sports in nine states. Data were analyzed with Spearman's rank-order correlation. Major correlations with CRP, LBP; MBL2; A1AG1, and SAA1 were found. The pairs CRP-SAA1 and CRP-LBP appeared with a robust positive correlation. Other pairs, LBP-SAA1; A1AG1-CRP; A1AG1-SAA1; A1AG1-MBL, and A1AG1-LBP, showed a broader correlation across the sports. The protein-protein interaction map revealed 1500 interactions with 44 core proteins, 30 of them linked to immune system processing. We propose that the inflammation follow-up in exercise can provide knowledge for internal cargo management in training, competition, recovery, doping control, and a deeper understanding of health and disease.

Sportomics suggests that albuminuria is a sensitive biomarker of hydration in cross combat.

We have been using sportomics to understand hypermetabolic stress. Cross Combat (CCombat) has recently been initiated as a high-intensity functional training method inspired by CrossFit. We used a CCombat session to induce metabolic stress and evaluated its effects on hydration and kidney function. Blood samples were collected from 16 elite-level professional male athletes engaged in training sessions over a 96-h protocol. Blood myoglobin increased by ~ 3.5-fold (119 ± 21 to 369 ± 62 nmol/L; p = .001) in response to the protocol, returning to the pre-exercise level within 48 h. Furthermore, D-dimer levels increased from 6.5 ± 0.6 to 79.4 ± 21.3 µmol/L (p < .001) in response to exercise decreasing during recovery with high variability among the studied athletes. Albuminemia and creatininemia increased ~ 10% and cystatin C increased ~ 240% (1.7 ± 0.1 to 5.7 ± 0.5 mg/L; p < .001; effect size = 2.4) in response to the protocol. We measured albuminuria (HuA) to assess kidney permeability to albumin caused by exercise. HuA increased ~ 16-fold (0.16 ± 0.03 to 2.47 ± 0.41 µmol/L; p < .001; effect size = 1.4) in response to exercise, dropping and reaching basal levels during 48 h. Here, we suggest that microalbuminuria can be used as an early, sensitive, easy, and inexpensive biomarker to evaluate hydration status changes during intensive exercise, decreasing chronic impairment in renal function.

Antimicrobial, antioxidant, volatile and phenolic profiles of cabbage-stalk and pineapple-crown flour revealed by GC-MS and UPLC-MSE.

Unconventional parts of vegetables represent a rich source of health-promoting phytochemicals. The phenolic profile of cabbage-stalk flour (CSF), pineapple-crown flour (PCF), and their essential oils were characterized via UPLC-ESI-QTOF-MSE and GC-FID/MS. Antimicrobial activity was tested against five strains, and antioxidant activities were determined in free and bound extracts. Globally, 177 phenolics were tentatively identified in PCF (major p-coumaric acid, ferulic acid, and 4-hydroxybenzaldehyde) and 56 in CSF (major chlorogenicacid, quercetin 3-O-glucuronide, and p-coumaric acid). PCF exhibited a distinguished profile (lignans, stilbenes) and antioxidant capacity, especially in bound extracts (1.3 g GAE.100 g-1; 0.6 g catechin eq.100 g-1; DPPH: 244.7; ABTS: 467.8; FRAP: 762.6 µg TE.g-1, ORAC: 40.9 mg TE.g-1). The main classes of volatile compounds were fatty acids, their esters, and terpenes in CSF (30) and PCF (41). A comprehensive metabolomic approach revealed CSF and PCF as a promising source of PC, showing great antioxidant and discrete antimicrobial activities.

Precision multiparameter tracking of inflammation on timescales of hours to years using serial dried blood spots.

Aim: High-frequency longitudinal tracking of inflammation using dried blood microsamples provides a new window for personalized monitoring of infections, chronic inflammatory disease and clinical trials of anti-inflammatory drugs. Results/methodology: Using 1662 dried blood spot samples collected by 16 subjects over periods of weeks to years, we studied the behavior of 12 acute phase response and related proteins in inflammation events correlated with infection, vaccination, surgery, intense exercise and Crohn's disease. Proteins were measured using SISCAPA mass spectrometry and normalized to constant plasma volume using low-variance proteins, generating high precision within-person biomarker trajectories with well-characterized personal baselines. Discussion/conclusion: The results shed new light on the dynamic regulation of APR responses, offering a new approach to visualization of multidimensional inflammation trajectories.

Byproduct Generated During the Elaboration Process of Isotonic Beverage as a Natural Source of Bioactive Compounds.

Agro-industrial byproducts are considered good sources of macronutrients and phytochemicals. Fruit and vegetable residues (FVR), obtained after the production of an isotonic beverage, have previously been characterized containing 80% insoluble dietary fibers from total fibers (48.4%), 26% available carbohydrates, 9.5% proteins and 5% lipids. Nevertheless, fruit and vegetables provide phytochemicals which have been related to human health such as phenolic compounds. The loss of specific compounds over the production process is related to their partitioning between fruit and vegetables and byproducts. However, phenolic profile of FVR remains unknown. This work is focused on the evaluation of FVR as a natural source of these bioactive compounds. For this purpose, pressurized liquid extraction (PLE) has been proposed as extraction technique for recovering phenolic compounds from FVR. The experimental variables were temperature and percentage of solvent (ethanol and water). Phenolic compounds extracts were characterized by UPLC-ESI-Q-TOF-MS and a discussion about phenolic and macronutrient interactions was established. Globally, 88 compounds were tentatively identified: phenolic acids (28), flavonoids (32), and other polyphenols (28). The PLE conditions applied yielded different breaking matrix-analyte interactions leading to an increase in the number of compounds. The highest phenolic acids content was achieved with high temperature while lower temperatures were more efficient in extracting flavonoid. By establishing the phenolics profile in food byproducts such as FVR, it is possible to more effectively apply these byproducts as nutraceutical, food or pharmaceutical ingredients. PRACTICAL APPLICATION: Flow diagram of bioactive compounds recovering from isotonic beverage byproduct is proposed using pressurized liquid extraction. The plant-bioactives mechanism relies on fruit and vegetable byproducts changes under different extraction conditions. The obtained extracts can most effectively be applied as nutraceuticals or as ingredients in food or pharmaceutical inputs.

Comparative metabolomic responses to gibberellic acid and 6-benzylaminopurine in Cunila menthoides Benth. (Lamiaceae): a contribution to understand the metabolic pathways.

KEY MESSAGE: Gibberellic acid elicited synthesis of many phenols from different classes and enhanced production of sesquiterpenoids, polyterpenoids, steroids and monoterpenoids compared to control and 6-benzylaminopurine. Little is known about the effects of 6-benzylaminopurine (BA) and gibberellic acid (GA3) on the synthesis of secondary metabolites in species of Lamiaceae. In this study, for the first time, the profile of secondary metabolites in plantlets of Cunila menthoides was characterized, using UPLC-ESI-Qq-oaTOF-MS. Ninety metabolites were identified, including polyphenols and terpenes. BA down-regulated most of the identified molecules in relation to GA3 and MS0 (control). The results showed that GA3 elicited synthesis of many phenols from different classes, and seemed to play a major role in the shikimate pathway in relation to BA. GA3 enhanced production of sesquiterpenoids, polyterpenoids, steroids and monoterpenoids compared to MS0 and BA, and also seemed to positively influence the MEP/DOXP and MVA pathways. These data show the most comprehensive metabolomic profile of Cunila menthoides to date, and the effects of BA and GA3 on the synthesis of secondary metabolites, modulating quantitative aspects of metabolism in Lamiaceae.

Sportomics: building a new concept in metabolic studies and exercise science.

For more than a decade, we have used alternative approaches to understand metabolic responses to physical stress. In addition to classic laboratory studies (cell and animal models), we have used elite athletes and sports to examine metabolic stress. Our central question involves the ability of the body to protect the central nervous system from high and toxic ammonemia during acute and chronic exercise. Information about this problem can aid in understanding important signaling pathways, which may yield better ways to protect people who suffer from diseases that lead to hyperammonemia, such as liver failure, or to hypermetabolic states, such as cancer or thermal injury. We proposed a Sportomics approach to mimic the real challenges and conditions that are faced during sports training and competition. Sportomics is non-hypothesis-driven research on an individual's metabolite changes during sports and exercise. It is similar to metabolomics and other "-omics" approaches, but Sportomics focuses on sports as a metabolic challenge. Our study is holistic and top-down; we treat the data systematically and have generated a large computer-searchable database. We also propose that in-field metabolic analyses are important for understanding, supporting and training elite athletes. In this review, we discuss Sportomics history, problems, benefits and results. We included different weather conditions, such as temperature, wind and humidity, and diverse metabolic responses due to uneven sleep and eating behaviors near the time of the experiment. We are currently generating databases as well as data-mining principles and procedures to improve metabolomics and proteomics studies as well as adding genomics and transcriptomics studies to the Sportomics approach. We believe that this approach can fill a methodological gap between systems biology and translational medicine similar as a bench to the field approach.

Enhanced transduction of photonic crystal dye lasers for gas sensing via swelling polymer film.

We present the enhanced transduction of a photonic crystal dye laser for gas sensing via deposition of an additional swelling polymer film. Device operation involves swelling of the polymer film during exposure to specific gases, leading to a change in total effective refractive index. Experimental results show an enhancement of 16.09 dB in sensing ethanol vapor after deposition of a polystyrene film. We verify different responses of the polystyrene film when exposed to either ethanol vapor or increased humidity, indicating selectivity. The concept is generic and, in principle, straightforward in its application to other intracavity-based detection schemes to enable gas sensing.

Chemical-genetic inhibition of a sensitized mutant myosin Vb demonstrates a role in peripheral-pericentriolar membrane traffic.

Selective, in situ inhibition of individual unconventional myosins is a powerful approach to determine their specific physiological functions. Here, we report the engineering of a myosin Vb mutant that still hydrolyzes ATP, yet is selectively sensitized to an N(6)-substituted ADP analog that inhibits its activity, causing it to remain tightly bound to actin. Inhibition of the sensitized mutant causes inhibition of accumulation of transferrin in the cytoplasm and increases levels of plasma-membrane transferrin receptor, suggesting that myosin Vb functions in traffic between peripheral and pericentrosomal compartments.

Dimethyl sulphoxide enhances the effects of P(i) in myofibrils and inhibits the activity of rabbit skeletal muscle contractile proteins.

In the catalytic cycle of skeletal muscle, myosin alternates between strongly and weakly bound cross-bridges, with the latter contributing little to sustained tension. Here we describe the action of DMSO, an organic solvent that appears to increase the population of weakly bound cross-bridges that accumulate after the binding of ATP, but before P(i) release. DMSO (5-30%, v/v) reversibly inhibits tension and ATP hydrolysis in vertebrate skeletal muscle myofibrils, and decreases the speed of unregulated F-actin in an in vitro motility assay with heavy meromyosin. In solution, controls for enzyme activity and intrinsic tryptophan fluorescence of myosin subfragment 1 (S1) in the presence of different cations indicate that structural changes attributable to DMSO are small and reversible, and do not involve unfolding. Since DMSO depresses S1 and acto-S1 MgATPase activities in the same proportions, without altering acto-S1 affinity, the principal DMSO target apparently lies within the catalytic cycle rather than with actin-myosin binding. Inhibition by DMSO in myofibrils is the same in the presence or the absence of Ca(2+) and regulatory proteins, in contrast with the effects of ethylene glycol, and the Ca(2+) sensitivity of isometric tension is slightly decreased by DMSO. The apparent affinity for P(i) is enhanced markedly by DMSO (and to a lesser extent by ethylene glycol) in skinned fibres, suggesting that DMSO stabilizes cross-bridges that have ADP.P(i) or ATP bound to them.

Calcium-induced quenching of intrinsic fluorescence in brain myosin V is linked to dissociation of calmodulin light chains.

Myosin V isolated from chick brain (BM V) is a multimeric protein of about 640 kDa consisting of two intertwined heavy chains of 212 kDa and multiple light chains of 10 to 20 kDa. A distinctive feature of the heavy chain is an extended neck region with six consensus IQ sites for the binding of calmodulin (CaM) and myosin light chains. The actin-activated MgATPase has been shown to require >/=1 microM Ca2+ for full activity, and evidence points to a myosin-linked regulatory system where the CaM light chains participate as modulators for the Ca2+ signal. Still, the precise mechanism of Ca2+ regulation remains unknown. In the present study we have used the intrinsic tryptophan fluorescence of native BM V to monitor conformational changes of BM V induced by Ca2+, and we relate these changes to CaM dissociation from the BM V molecule. The fluorescence intensity decreases approximately 17% upon addition of sub-micromolar concentrations of Ca2+ (K0.5 = 0.038 microM). This decrease in fluorescence, which is dominated by a conformational change in the heavy chain, can be reversed by addition of 1, 2-di(2-aminoethoxy)ethane-N,N,N',N'tetraacetic acid (EGTA) followed by an excess of CaM, but not by addition of EGTA alone. Gel filtration of native BM V using HPLC shows that CaM is partially dissociated from the heavy chain in EGTA and dissociates further upon addition of sub-micromolar concentrations of Ca2+. These observations suggest that the affinity of CaM for at least one of the IQ sites on the BM V heavy chain decreases with Ca2+ and that the Ca2+ concentration required for this effect is lower than that needed to activate acto-BM V. Using a cosedimentation assay in the presence of actin, we also observe partial dissociation of CaM when Ca2+ is absent, but now the addition of Ca2+ has a biphasic effect: sub-micromolar Ca2+ concentrations lead to reassociation of CaM with the heavy chain, followed by dissociation when Ca2+ exceeds 5-10 microM. Thus, the binding of CaM to BM V is affected by both actin and Ca2+.