Sportomics Analyses of the Exercise-Induced Impact on Amino Acid Metabolism and Acute-Phase Protein Kinetics in Female Olympic Athletes.

BACKGROUND: Exercise can be used as a model to understand immunometabolism. Biological data on elite athletes are limited, especially for female athletes, including relevant data on acute-phase proteins and amino acid metabolism. METHODS: We analyzed acute-phase proteins and amino acids collected at South American, Pan-American, and Olympic Games for 16 Olympic sports. We compared female and male elite athletes (447 vs. 990 samples) across four states (fasting, pre-exercise, post-exercise, and resting) to understand sex-specific immunometabolic responses in elite athletes. RESULTS: Considering all states and sports, we found that elite female athletes exhibited higher concentrations of C-reactive protein, lipopolysaccharide-binding protein, myeloperoxidase, haptoglobin, and IGF1, with ratios ranging from 1.2 to 2.0 (p < 0.001). Women exhibited lower concentrations of most amino acids, except for glutamate and alanine. Although almost 30% lower in women, branched-chain amino acids (BCAAs) showed a similar pattern in all states (ρ ≥ 0.9; p < 0.001), while aromatic amino acids (AAAs) showed higher consumption during exercise in women. CONCLUSION: We established sex dimorphism in elite athletes' metabolic and inflammatory responses during training and competition. Our data suggest that female athletes present a lower amino acid response towards central fatigue development than male athletes. Understanding these differences can lead to insights into sex-related immuno-metabolic responses in sports or other inflammatory conditions.

The Modulation of Septic Shock: A Proteomic Approach.

Sepsis poses a significant challenge due its lethality, involving multiple organ dysfunction and impaired immune responses. Among several factors affecting sepsis, monocytes play a crucial role; however, their phenotype, proteomic profile, and function in septic shock remain unclear. Our aim was to fully characterize the subpopulations and proteomic profiles of monocytes seen in septic shock cases and discuss their possible impact on the disease. Peripheral blood monocyte subpopulations were phenotype based on CD14/CD16 expression by flow cytometry, and proteins were extracted from the monocytes of individuals with septic shock and healthy controls to identify changes in the global protein expression in these cells. Analysis using 2D-nanoUPLC-UDMSE identified 67 differentially expressed proteins in shock patients compared to controls, in which 44 were upregulated and 23 downregulated. These proteins are involved in monocyte reprogramming, immune dysfunction, severe hypotension, hypo-responsiveness to vasoconstrictors, vasodilation, endothelial dysfunction, vascular injury, and blood clotting, elucidating the disease severity and therapeutic challenges of septic shock. This study identified critical biological targets in monocytes that could serve as potential biomarkers for the diagnosis, prognosis, and treatment of septic shock, providing new insights into the pathophysiology of the disease.

Caffeine Boosts Weight-Lifting Performance in Rats: A Pilot Study.

Caffeine is a well-described ergogenic aid used to enhance athletic performance. Using animal models can greatly increase our understanding of caffeine's mechanisms in performance. Here, we adapted an animal weight-lifting exercise model to demonstrate caffeine's ergogenic effect in rats. Male Wistar rats (315 ± 35 g) were randomly divided into two groups: one group received 5 mg·kg-1 of caffeine (0.5 mL; CEx; n = 5) and the other 0.9% NaCl (0.5 mL; PEx; n = 4) through an orogastric probe (gavage) one hour before exercise. Weight-lifting exercise sessions were performed over three subsequent days, and the number of complete squats performed was counted. Analyses of the area under the curve in all three experiments showed that the CEx group responded more to stimuli, performing more squats (1.7-, 2.0-, and 1.6-fold; p < 0.05) than the control group did. These three days' data were analyzed to better understand the cumulative effect of this exercise, and a hyperbolic curve was fitted to these data. Data fitting from the caffeine-supplemented group, CEx, also showed larger Smax and Kd (2.3-fold and 1.6-fold, respectively) than the PEx group did. Our study demonstrated an acute ergogenic effect of caffeine in an animal weight-lifting exercise model for the first time, suggesting potential avenues for future research.

From Microcosm to Macrocosm: The -Omics, Multiomics, and Sportomics Approaches in Exercise and Sports.

This article explores the progressive integration of -omics methods, including genomics, metabolomics, and proteomics, into sports research, highlighting the development of the concept of "sportomics." We discuss how sportomics can be used to comprehend the multilevel metabolism during exercise in real-life conditions faced by athletes, enabling potential personalized interventions to improve performance and recovery and reduce injuries, all with a minimally invasive approach and reduced time. Sportomics may also support highly personalized investigations, including the implementation of n-of-1 clinical trials and the curation of extensive datasets through long-term follow-up of athletes, enabling tailored interventions for athletes based on their unique physiological responses to different conditions. Beyond its immediate sport-related applications, we delve into the potential of utilizing the sportomics approach to translate Big Data regarding top-level athletes into studying different human diseases, especially with nontargeted analysis. Furthermore, we present how the amalgamation of bioinformatics, artificial intelligence, and integrative computational analysis aids in investigating biochemical pathways, and facilitates the search for various biomarkers. We also highlight how sportomics can offer relevant information about doping control analysis. Overall, sportomics offers a comprehensive approach providing novel insights into human metabolism during metabolic stress, leveraging cutting-edge systems science techniques and technologies.

A sportomics soccer investigation unveils an exercise-induced shift in tyrosine metabolism leading to hawkinsinuria.

Tyrosine metabolism has an intense role in the synthesis of neurotransmitters. Our study used an untargeted, sportomics-based analysis of urine samples to investigate changes in metabolism during a soccer match in 30 male junior professional soccer players. Samples were collected before and after the match and analyzed using liquid chromatography and mass spectrometry. Results showed significant changes in tyrosine metabolism. Exercise caused a downregulation of the homogentisate metabolites 4-maleylacetoacetate and succinylacetone to 20% (p = 4.69E-5) and 16% (p = 4.25E-14), respectively. 4-Hydroxyphenylpyruvate, a homogentisate precursor, was found to be upregulated by 26% (p = 7.20E-3). The concentration of hawkinsin and its metabolite 4-hydroxycyclohexyl acetate increased ~six-fold (p = 1.49E-6 and p = 9.81E-6, respectively). Different DOPA metabolism pathways were also affected by exercise. DOPA and dopaquinone increased four-to six-fold (p = 5.62E-14 and p = 4.98E-13, respectively). 3-Methoxytyrosine, indole-5,6-quinone, and melanin were downregulated from 1 to 25%, as were dopamine and tyramine (decreasing to up to 5% or 80%; p= 5.62E-14 and p = 2.47E-2, respectively). Blood TCO2 decreased as well as urinary glutathione and glutamate (40% and 10% respectively) associated with a two-fold increase in pyroglutamate. Our study found unexpected similarities between exercise-induced changes in metabolism and the inherited disorder Hawkinsinuria, suggesting a possible transient condition called exercise-induced hawkinsinuria (EIh). Additionally, our research suggests changes in DOPA pathways may be involved. Our findings suggest that soccer exercise could be used as a model to search for potential countermeasures in Hawkinsinuria and other tyrosine metabolism disorders.

A burst of fenoterol excretion during the recovery of a weight loss protocol.

Fenoterol is a sympathomimetic ß2 receptor agonist primarily used as a bronchodilator. Due to its sympathomimetic actions, the World Anti-Doping Agency (WADA) has banned it. Multiple acute weight loss protocols (WLP) are used by Olympic athletes for sports that segregate athletes by weight; these generally involve caloric and water deprivation combined with heat exposure. Athletes use WLP before weigh-in, then transition to different body acute weight regain protocols (WRP) before competitions. Here, we studied the pharmacokinetics of fenoterol under WLP conditions: energetic dietary restriction, decreased water intake, and exposure to a dry sauna (80 ± 2 °C), followed by a WRP. Five elite-level female judo athletes participated in the study. Four received fenoterol (200 µg; n = 2 or 400 µg; n = 2), while one was a control receiving placebo under identical conditions. We measured excretion of the fenoterol parent molecule and presented qualitative data of its sulfated metabolite using QqQ tandem quadrupole mass spectrometry for 118 h. The fenoterol parent appeared earlier in urine than did its conjugated metabolite; excretion profiles were similar among all subjects. The centers of mass for fenoterol parent curves were (time, fenoterol): athlete A (10.9, 7.3); athlete B (9.2, 27.3); athlete C (8.5, 6.9); athlete D (9.7, 5.0). After initiating WRP, we observed a burst in urinary fenoterol excretion once in complete decay. This trend was observed for all four athletes who received fenoterol. Our results suggest that during hypohydration, some of the unmetabolized fenoterol accumulates in tissues, then is released during rehydration. These findings can be important for detecting fenoterol use in athletes.

Dietary higenamine from Annonaceae family fruits as a possible source of unintentional doping.

Members of the genus Aconitum have been used for millennia, both as poisons and medicines, in Eastern culture. Higenamine has non-selective beta-agonist effects, activating both ß1 and ß2 adrenoreceptors, and is present in a variety of plants. The World Anti-Doping Agency has banned Higenamine both in competition and out of competition. Due to the common uses of higenamine in Brazilian culture, both as medicine and food, we studied the urinary concentrations of higenamine after the consumption of fruits of the Annona genus. We evaluated whether the ingestion of these fruits has the potential to cause anti-doping code violations. We measured higenamine concentrations for a 72 h period in the urine of ten healthy, physically active males (age 20-30; weight 70-80 kg; not consuming supplements or medications) after eating a unique meal containing fruits. Fruit consumption ranges were: Carica papaya (control) 348 ± 98 g; A. muricata 450 ± 282 g; and A. squamosa 314 ± 60 g. (all mean± SD). Higenamine was measured using ultra-performance liquid chromatography coupled with electrospray-tandem mass spectrometry. The appearance of urinary higenamine occurred within the first 12 h after eating A. muricata (n = 3), and the maximum concentration found was 1.9 ng/mL. The ingestion of A. squamosa has also been shown to cause higenamine urinary excretion. The elimination kinetics of the subjects who ingested A. squamosa (n = 4) were different from each other. After ingestion of the control fruit, C. papaya, we detected no higenamine in the urine of any participants (n = 3). Although the kinetics varied by individuals and fruits, A. muricata ingestion produced higher higenamine excretion; however, the A. squamosa portion weighed ∼66 % of the A. muricata portion. We conclude that eating Annonaceae family fruits cause detectable higenamine excretion. Conversely, single ingestion did not reach the WADA's threshold to cause adverse analytical findings.

Metabolomic analysis and ecofriendly enrichment of sunflower meal extract.

BACKGROUND: The presence of phenolic compounds in sunflower is well reported in the literature; however, knowledge is scarce when it comes to the composition of other secondary metabolites in this species and their by-products. This work evaluated, for the first time, the phytochemical composition of sunflower meal produced in Brazil. A combination of mixture design and central composite rotatable design 23 models was then applied to maximize the recovery of bioactive compounds using ecologically friendly solvents and concentrating by applying activated carbon, a sustainable adsorbent. The product of this extraction-concentration was also evaluated by an untargeted metabolomic approach using ultra-performance liquid chromatography coupled to mass spectrometry. RESULTS: A diverse and abundant profile of phenolic compounds was obtained from Brazilian sunflower meal: in total, 51 natural products were tentatively identified, 35 of which for the first time in sunflower. The sorption capacity of the activated charcoal, in the optimized process conditions, was effective in the separation and concentration of minority secondary metabolites. The ecofriendly extract proved to be enriched in plumberoside, p-coumaric acid, and alkaloids. CONCLUSIONS: Investigation of the phytochemical profile of sunflower meal produced in Brazil pointed to several secondary metabolites reported for the first time in sunflower samples, including phenolic compounds, alkaloids, and terpenes. The use of activated charcoal in an alkaline medium as an adsorbent for the concentration of these phytochemicals, from an aqueous extract, generated a potentially cost-effective, ecofriendly extract, enriched in minor metabolites, indicating a possible innovative way to selectively obtain these compounds from sunflower meal. © 2022 Society of Chemical Industry.

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.

Effect of extrusion and turmeric addition on phenolic compounds and kafirin properties in tannin and tannin-free sorghum.

Sorghum is a potential substitute for corn/wheat in cereal-based extruded products. Despite agronomic advantages and its rich diversity of phenolic compounds, sorghum kafirins group together and form complex with tannins, leading to a low digestibility. Phenolic content/profile by UPLC-ESI-QTOF-MSE and kafirins polymerization by SE-HPLC were evaluated in wholemeal sorghum extrudates; tannin-rich (#SC319) and tannin-free (#BRS330) genotypes with/without turmeric powder. Total phenolic, proantocyanidin and flavonoid contents were strongly correlated with antioxidant capacity (r > 0.9, p < 0.05). Extrusion increased free (+60%) and decreased bound phenolics (-40%) in #SC319, but reduced both (-40%; -90%, respectively) in #BRS330, which presented lower abundance after extrusion. Turmeric addition did not significantly impact antioxidant activity, phenolic content and profile and kafirins profile. Tannins presence/absence impacted phenolic profiles and polymerization of kafirins which appears related to the thermoplastic process. The extrusion improved proteins solubility and can positively enhance their digestibility (phenolic compounds-proteins interactions), making more accessible to proteolysis in sorghum extrudates.

NanoUPLC-MSE reveals differential abundance of gluten proteins in wheat flours of different technological qualities.

Gluten proteins contribute to the rheological properties of dough. Mass spectrometric techniques help to understand the contribution of these proteins to the quality of the end product. This work aimed to apply modern proteomic techniques to characterize and provide a better understanding of gluten proteins in wheat flours of different technological qualities. Nine Brazilian wheat flours (Triticum aestivum) classified by rheological gluten force were used to extract the proteins. Extracts were pooled together by technological qualities in low (LW), medium (MD), and superior (SP). Peptides were analyzed by nanoUPLC and mass spectrometry multiplex method (MSE). Collectively, 3545 peptides and 1297 proteins were identified, and 116 proteins were found differentially abundant. Low molecular weight glutenin subunits (LMW-GS) were found up-regulated only in SP samples. Proteins related to wheat grain hardness, such as puroindoline-A, were found in significant concentration in LW samples. After domain prediction, LW presented a different pattern with a lower abundance of functional domains, and SP presented chaperones, known to be involved in adequate folding of the storage proteins. NanoUPLC-MSE was efficient in analyzing and distinguishing the proteomic pattern of wheat flours from different qualities, pointing out the differentially abundant gluten proteins and providing a better understanding of wheat flour quality. SIGNIFICANCE: Common wheat is one of the most important staple food sources in the world. The improvement and comprehension of wheat quality has been a major objective of plant breeders and cereal chemists. Our findings highlighted the application of a modern proteomic approach to obtain a better understanding of the impact of gluten proteins on the technological quality of different wheat flours. The obtained data revealed different abundances of wheat quality-related proteins in superior quality flours when compared with samples of low rheological properties. In addition, multivariate statistical analysis clearly distinguished the flours of different qualities. This work contributes to the consolidation of research in the field of wheat technological quality.

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.

Keto analogues and amino acid supplementation and its effects on ammonaemia during extenuating endurance exercise in ketogenic diet-fed rats.

Keto analogues and amino acids (KAAA) supplementation can reduce blood ammonia concentrations in athletes undergoing high-intensity exercise under both ketogenic and thermoneutral conditions. This study evaluated the acute effects of KAAA supplementation on ammonia metabolism during extenuating endurance exercise in rats fed a ketogenic diet. In all, eighty male Fischer rats at 90 d of age were divided into eight groups, and some were trained using a swimming endurance protocol. A ketogenic diet supplemented with keto analogues was administered for 10 d. Administration of the ketogenic diet ended 3 d before the exhaustion test (extenuating endurance exercise). A ketogenic diet plus KAAA supplementation and extenuating endurance exercise (trained ketogenic diet supplemented with KAAA (TKKa)) increased blood ammonia concentrations by approximately 50 % compared with the control diet (trained control diet supplemented with KAAA (TCKa)) and similar training (effect size=1·33; statistical power=0·50). The KAAA supplementation reduced blood urea concentrations by 4 and 18 % in the control and ketogenic diet groups, respectively, compared with the groups fed the same diets without supplementation. The trained groups had 60 % lower blood urate concentrations after TCKa treatment than after TKKa treatment. Our results suggest that KAAA supplementation can reduce blood ammonia concentrations after extenuating endurance exercise in rats fed a balanced diet but not in rats fed a ketogenic diet.

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.

Differential expression of albumins and globulins of wheat flours of different technological qualities revealed by nanoUPLC-UDMSE.

Soluble proteins were extracted from common wheat flour obtained from nine cultivars of different qualities and were analyzed by nanoUPLC and Ultra Definition Mass Spectrometry (UDMSE) label-free quantitative approach. Collectively, 5894 proteins were identified and quantified with 8 peptides/protein. A total of 414 proteins were found differentially expressed with 85% proteins not yet described in the literature, according to their biological function. Quality-related proteins, such as puroindolines and chaperones, notably involved in the gluten-protein folding process, were up-regulated in superior (SP) and medium (MD) flours qualities and down-regulated in the low (LW) ones. Domains related to the chaperones activity were also found up-regulated in SP and MD and down-regulated in LW. In this work, nanoUPLC-UDMSE analysis was effective to identify and quantify metabolic proteins, allowing a clear distinction of the wheat flours and providing new perspectives for evaluating the technological quality based on the low abundant proteins of wheat endosperm.

Nanoscale Plasmonic V-Groove Waveguides for the Interrogation of Single Fluorescent Bacterial Cells.

We experimentally demonstrate the interrogation of an individual Escherichia coli cell using a nanoscale plasmonic V-groove waveguide. Several different configurations were studied. The first involved the excitation of the cell in a liquid environment because it flows on top of the waveguide nanocoupler, while the obtained fluorescence is coupled into the waveguide and collected at the other nanocoupler. The other two configurations involved the positioning of the bacterium within the nanoscale waveguide and its excitation in a dry environment either directly from the top or through waveguide modes. This is achieved by taking advantage of the waveguide properties not only for light guiding but also as a mechanical tool for trapping the bacteria within the V-grooves. The obtained results are supported by a set of numerical simulations, shedding more light on the mechanism of excitation. This demonstration paves the way for the construction of an efficient bioplasmonic chip for diverse cell-based sensing applications.