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The aim of this study was to determine the effectiveness of a 7-day oral supplementation with branched-chain amino acids (BCAA) to prevent muscle damage during a marathon. Forty-six experienced runners were randomly divided into two groups, one with BCAA supplementation (n = 25, supplemented with 5 g day⁻¹of powdered 1:0.5:0.5 leucine:isoleucine:valine, during the 7 days prior to the competition) and the other as a control group (n = 21, supplemented with an isocaloric placebo). Before the marathon race and within 3 min of finishing, leg muscle power was measured with a maximal countermovement jump and a urine sample was obtained. During the race, running pace was measured by means of a time-chip. Myoglobin concentration was determined in the urine samples as an indirect marker of muscle damage. A visual analog scale (0–10 points) was used to assess leg muscle pain during the race. In the BCAA group, the mean running pace during the marathon was similar to the control group (3.3 ± 0.4 vs. 3.3 ± 0.5 m s⁻¹, respectively, 0.98). The pre- to post-race reduction in muscle power was similar in both BCAA and control groups (−23.0 ± 16.1 vs. −17.3 ± 13.8 %, P = 0.13). Post-race urine myoglobin concentration was similar in both BCAA and control groups (5.4 ± 7.5 vs. 4.5 ± 8.6 μg mL⁻¹, P = 0.70). Finally, there were no differences between groups in the perceived muscle pain during the race (6 ± 1 vs. 5 ± 1 points, P = 0.80). A 7-day supplementation of BCAA (5 g day⁻¹) did not increase the running performance during a marathon. Furthermore, BCAA supplementation was ineffective to prevent muscle power loss, muscle damage or perceived muscle pain during a marathon race.

Intensive physical exercise may cause muscular injury and increase oxidative stress. The purpose of this study was to examine the effect of an antioxidant, coenzyme Q10 (CoQ10), on muscular injury and oxidative stress during exercise training. Eighteen male students, all elite Japanese kendo athletes, were randomly assigned to either a CoQ10 group (n 10) or a placebo group (n 8) in a double-blind manner. Subjects in the CoQ10 group took 300 mg CoQ10 per d for 20 d, while subjects in the placebo group took the same dosage of a placebo. All subjects practised kendo 5·5 h per d for 6 d during the experimental period. Blood samples were taken 2 weeks before, during (1 d, 3 d, 5 d) and 1 week after the training. Serum creatine kinase (CK) activity and myoglobin (Mb) concentration significantly increased in both groups (at 3 d and 5 d). Serum CK (at 3 d), Mb (at 3 d) and lipid peroxide (at 3 d and 5 d) of the CoQ10 group were lower than those of the placebo group. The leucocyte counts in the placebo group significantly increased (at 3 d) and neutrophils significantly increased in both groups (at 3 d and 5 d). Serum scavenging activity against superoxide anion did not change in either group. These results indicate that CoQ10 supplementation reduced exercise-induced muscular injury in athletes.

The aim of the present study was to gain better insight into the mechanisms underpinning the sigmoid pattern of deoxy[Hb + Mb] during incremental exercise by assessing the changes in the profile following prior high-intensity exercise. Ten physically active students performed two incremental ramp (25 W min −1 ) exercises (AL and LL, respectively) preceded on one occasion by incremental arm (10 W min −1 ) and on another occasion by incremental leg exercise (25 W min −1 ), which served as the reference test (RT). Deoxy[Hb + Mb] was measured by means of near-infrared spectroscopy and surface EMG was recorded at the Vastus Lateralis throughout the exercises. Deoxy[Hb + Mb], integrated EMG and Median Power Frequency (MdPF) were expressed as a function of work rate (W) and compared between the exercises. During RT and AL deoxy[Hb + Mb] followed a sigmoid increase as a function of work rate. However, during LL deoxy[Hb + Mb] increased immediately from the onset of the ramp exercise and thus no longer followed a sigmoid pattern. This different pattern in deoxy[Hb + Mb] was accompanied by a steeper slope of the iEMG/ W -relationship below the GET (LL: 0.89 ± 0.11% W −1 ; RT: 0.74 ± 0.08% W −1 ; AL: 0.72 ± 0.10% W −1 ) and a more pronounced decrease in MdPF in LL (17.2 ± 4.5%) compared to RT (5.0 ± 2.1%) and AL (3.9 ± 3.2%). It was observed that the sigmoid pattern of deoxy[Hb + Mb] was disturbed when the ramp exercise was preceded by priming leg exercise. Since the differences in deoxy[Hb + Mb] were accompanied by differences in EMG it can be suggested that muscle fibre recruitment is an important underlying mechanism for the pattern of deoxy[Hb + Mb] during ramp exercise.

Enzymatic digestion for protein sequencing usually requires much time, and does not always result in high sequence coverage. Here we report the use of aqueous microdroplets to accelerate enzymatic reactions and, in particular, to improve protein sequencing. When a room temperature aqueous solution containing 10 µM myoglobin and 5 µg mL −1 trypsin is electrosonically sprayed (−3 kV) from a homemade setup to produce tiny (∼9 µm) microdroplets, we obtain 100% sequence coverage in less than 1 ms of digestion time, in sharp contrast to 60% coverage achieved by incubating the same solution at 37 °C for 14 h followed by analysis with a commercial electrospray ionization source that produces larger (∼60 µm) droplets. We also confirm the sequence of the therapeutic antibody trastuzumab (∼148 kDa), with a sequence coverage of 100% for light chains and 85% for heavy chains, demonstrating the practical utility of microdroplets in drug development. Mass spectrometry (MS)-based protein sequencing usually relies on in-solution proteolytic digestion, which is time-consuming and inefficient for certain proteins. Here, the authors achieve full protein sequence coverage in less than 1 ms by subjecting protein-protease mixtures to electrosonic spray ionization-MS.

Hemoglobin and myoglobin are widely responsible for oxygen transport and storage (see the Perspective by Rezende ). The ability of diving mammals to obtain enough oxygen to support extended dives and foraging is largely dependent on muscle myoglobin (Mb) content. Mirceta et al. (p. 1234192 ) found that in mammalian lineages with an aquatic or semiaquatic lifestyle, Mb net charge increases, which may represent an adaptation to inhibit self-association of Mb at high intracellular concentrations. Epistasis results from nonadditive genetic interactions and can affect phenotypic evolution. Natarajan et al. (p. 1324 ) found that epistatic interactions were able to explain the increased hemoglobin oxygen-binding affinity observed in deer mice populations at high altitude. In mammals, the offloading of oxygen from hemoglobin is facilitated by a reduction in the blood's pH, driven by metabolically produced CO 2 . However, in fish, a reduction in blood pH reduces oxygen carrying capacity of hemoglobin. Rummer et al. (p. 1327 ) implanted fiber optic oxygen sensors within the muscles of rainbow trout and found that elevated CO 2 levels in the water led to acidosis and elevated oxygen tensions. Increasing the number of charged amino acids allows for higher myoglobin concentrations in the muscles of diving mammals. [Also see Perspective by Rezende ] Extended breath-hold endurance enables the exploitation of the aquatic niche by numerous mammalian lineages and is accomplished by elevated body oxygen stores and adaptations that promote their economical use. However, little is known regarding the molecular and evolutionary underpinnings of the high muscle myoglobin concentration phenotype of divers. We used ancestral sequence reconstruction to trace the evolution of this oxygen-storing protein across a 130-species mammalian phylogeny and reveal an adaptive molecular signature of elevated myoglobin net surface charge in diving species that is mechanistically linked with maximal myoglobin concentration. This observation provides insights into the tempo and routes to enhanced dive capacity evolution within the ancestors of each major mammalian aquatic lineage and infers amphibious ancestries of echidnas, moles, hyraxes, and elephants, offering a fresh perspective on the evolution of this iconic respiratory pigment.

This study monitored the impact of seasonal feeding variability on meat quality traits in forage-fed Angus steers. A total of 38 steers were selected based on their diets during the last three months before slaughter: a hay-based winter diet (WIN), a spring fresh-forage-based diet (SPR), or a summer fresh-forage-based diet (SMM). Meat quality was assessed through fatty acid profiling, antioxidant capacity, myoglobin content, color stability, and lipid oxidation during refrigerated storage. Principal components and linear discriminant analysis were used to uncover, underlying patterns, and classify the observations based on the multivariate profile. Meat fatty acids showed limited variation across seasons, with differences restricted to minor polyunsaturated fatty acids. The n-6/n-3 ratio and other health-related lipid indices remained within recommended values despite the feeding season. Meat from SPR and SMM animals showed higher α - and γ -tocopherol levels and greater myoglobin content compared to WIN meat. Color parameters and lipid oxidation were moderately affected by diet and storage time, with better stability observed in meat from animals receiving fresh forage. Despite the subtle impact on individual quality traits, multivariate discriminant analysis effectively differentiated meat samples based on the feeding season, achieving 89.5% correct classification. The most discriminating variables included fatty acids (17:1 c 9 , 18:1 t 10, 22:0, 22:5 n-6), color attributes (b*), and oxidative markers (TBARS, metmyoglobin %). With the caution due to the nature of the study, these findings suggest that while single quality indicators may not strongly reflect dietary differences, their combination may provide a tool to trace seasonal feeding strategies in grass-fed beef production.

Water dispersible L-glutamic acid (Glu) functionalized cesium lead bromide perovskite quantum dots (CsPbBr 3 PQDs), namely CsPbBr 3 @Glu PQDs were synthesized and used for the fluorescence “turn-off” detection of myoglobin (Myo). The as-prepared CsPbBr 3 @Glu PQDs exhibited an exceptional photoluminescence quantum yield of 25% and displayed emission peak at 520 nm when excited at 380 nm. Interestingly, the fluorescence “turn-off” analytical approach was designed to detect Myo using CsPbBr 3 @Glu PQDs as a simple optical probe. The developed probe exhibited a wide linear range (0.1–25 µM) and a detection limit of 42.42 nM for Myo sensing. The CsPbBr 3 @Glu PQDs-based optical probe provides high ability to determine Myo in serum and plasma samples. Graphical Abstract

A label free electrochemical sensor based on pure titanium oxide and manganese (Mn)-doped titanium oxide (TiO2) nanoparticles are fabricated and characterized for the sensitive detection of myoglobin (Mb) levels to analyze the cardiovascular infarction. Pristine and Mn-doped TiO2 nanoparticles were synthesized via the sol-gel method and characterized in order to understand their structure, morphologies, composition and optical properties. The structural properties revealed that the pure- and doped-TiO2 nanoparticles possess different TiO2 planes. FTIR studies confirm the formation of metal oxide nanoparticles by exhibiting a well-defined peak in the range of 600–650 cm−1. The values of the optical band gap, estimated from UV-Vis spectroscopy, are decreased for the Mn-doped TiO2 nanoparticles. UV-Vis spectra in the presence of myoglobin (Mb) indicated interaction between the TiO2 nanoparticles and myoglobin. The SPE electrodes were then fabricated by printing powder film over the working electrode and tested for label-free electrochemical detection of myoglobin (Mb) in the concentration range of 0–15 nM Mb. The fabricated electrochemical sensor exhibited a high sensitivity of 100.40 μA-cm−2/nM with a lowest detection limit of 0.013 nM (0.22 ng/mL) and a response time of ≤10 ms for sample S3. An interference study with cyt-c and Human Serum Albumin (HSA) of the sensors show the selective response towards Mb in 1:1 mixture.

Forensic biochemistry has often relied on the vitreous humor as a matrix for toxicological investigations due to its stability and isolation from post-mortem redistribution processes. Recently, the scope of research has expanded to explore the vitreous humor as a medium reflecting systemic and pathological changes, particularly in its protein composition. This study delves into the detection and quantification of cardiac damage markers such as CK-MB and myoglobin in vitreous humor samples from 45 autopsy cases. For the first time, it demonstrates a statistically significant correlation between these markers and the perimortem agony interval (PAI), defined as the survival time before death. This discovery paves the way for innovative forensic applications, including the estimation of the PAI, a critical parameter for judicial and compensatory assessments. The findings underscore the potential of the vitreous humor as a diagnostic medium, opening new avenues for understanding the systemic dynamics of cardiac markers and the role of the blood–retinal barrier in post-mortem scenarios.

A nanohybrid mediated SERS substrate was prepared by in-situ synthesis and assembly of gold nanoparticles (AuNPs) on exfoliated nanosheets of tungsten disulfide (WS 2 ) to form plasmonic hotspots. The nanohybrid surface was functionalized with specific aptamers which imparted high selectivity for the cardiac marker myoglobin (Mb). The fabricated aptasensor was read by SERS using a 532 nm laser and demonstrated significant signal enhancement, and this allowed Mb to be determined in the 10 f. mL −1 to 0.1 μg mL −1 concentration range. The study presents an approach to synergistically exploit the unique chemical and electromagnetic properties of both WS 2 and AuNPs for many-fold enhancement of SERS signals. Graphical abstract Schematic presentation of a nanohybrid-mediated SERS substrate prepared by in-situ assembly of gold nanoparticles (AuNPs) reduced on exfoliated nanosheets of tungsten disulfide (WS 2 ) to form plasmonic hot spots. Specific aptamers immobilized on the SERS surface impart high sensitivity and selectivity for the cardiac marker myoglobin (Mb).