scholarly journals Metabolomic analysis of skeletal muscle before and after strenuous exercise to fatigue

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hajime Ohmura ◽  
Kazutaka Mukai ◽  
Yuji Takahashi ◽  
Toshiyuki Takahashi
Keyword(s):  
Skeletal Muscle ◽  
Tricarboxylic Acid Cycle ◽  
Maximal Oxygen Consumption ◽  
Incremental Exercise ◽  
Mass Spectrometry Analysis ◽  
Strenuous Exercise ◽  
Metabolomic Analysis ◽  
Exercise Tests ◽  
Spectrometry Analysis ◽  
Before And After

AbstractThoroughbreds have high maximal oxygen consumption and show hypoxemia and hypercapnia during intense exercise, suggesting that the peripheral environment in skeletal muscle may be severe. Changes in metabolites following extreme alterations in the muscle environment in horses after exercise may provide useful evidence. We compared the muscle metabolites before and after supramaximal exercise to fatigue in horses. Six well-trained horses ran until exhaustion in incremental exercise tests. Biopsy samples were obtained from the gluteus medius muscle before and immediately after exercise for capillary electrophoresis–mass spectrometry analysis. In the incremental exercise test, the total running time and speed of the last step were 10.4 ± 1.3 (mean ± standard deviation) min and 12.7 ± 0.5 m/s, respectively. Of 73 metabolites, 18 and 11 were significantly increased and decreased after exercise, respectively. The heat map of the hierarchical cluster analysis of muscle metabolites showed that changes in metabolites were clearly distinguishable before and after exercise. Strenuous exercise increased many metabolites in the glycolytic pathway and the tricarboxylic acid cycle in skeletal muscle. Targeted metabolomic analysis of skeletal muscle may clarify the intramuscular environment caused by exercise and explain the response of working muscles to strenuous exercise that induces hypoxemia and hypercapnia in Thoroughbred horses.

scholarly journals Novel Phototransformable Fluorescent Protein SAASoti with Unique Photochemical Properties

2019 ◽  
Vol 20 (14) ◽  
pp. 3399 ◽  
Author(s):  
Ilya D. Solovyev ◽  
Alexandra V. Gavshina ◽  
Alexander P. Savitsky
Keyword(s):  
Fluorescent Protein ◽  
Light Exposure ◽  
Mass Spectrometry Analysis ◽  
Fast Recovery ◽  
Spectrometry Analysis ◽  
Before And After ◽  
In The Wild ◽  
Photochemical Properties ◽  
Type Gene ◽  
Red Form

SAASoti is a unique fluorescent protein (FP) that combines properties of green-to-red photoconversion and reversible photoswitching (in its green state), without any amino acid substitutions in the wild type gene. In the present work, we investigated its ability to photoswitch between fluorescent red (‘on’) and dark (‘off’) states. Surprisingly, generated by 400 nm exposure, the red form of SAASoti (R1) does not exhibit any reversible photoswitching behavior under 550 nm illumination, while a combination of prior 470 nm and subsequent 400 nm irradiation led to the appearance of another—R2—form that can be partially photoswitched (550 nm) to the dark state, with a very fast recovery time. The phenomenon might be explained by chemical modification in the chromophore microenvironment during prior 470 nm exposure, and the resulting R2 SAASoti differs chemically from the R1 form. The suggestion is supported by the mass spectrometry analysis of the tryptic peptides before and after 470 nm light exposure, that revealed Met164 oxidation, as proceeds in another dual phototransformable FP, IrisFP.

Effect of short-term exercise training on insulin-stimulated PI 3-kinase activity in human skeletal muscle

1999 ◽  
Vol 277 (6) ◽  
pp. E1055-E1060 ◽  
Author(s):  
Joseph A. Houmard ◽  
Christopher D. Shaw ◽  
Matthew S. Hickey ◽  
Charles J. Tanner
Keyword(s):  
Skeletal Muscle ◽  
Insulin Sensitivity ◽  
Exercise Training ◽  
Kinase Activity ◽  
Vastus Lateralis ◽  
Maximal Oxygen Consumption ◽  
Exercise Bout ◽  
Phosphatidylinositol 3 Kinase ◽  
Insulin Signal Transduction ◽  
Before And After

The purpose of this study was to determine if the improvement in insulin sensitivity with exercise training is associated with enhanced phosphatidylinositol 3-kinase (PI 3-kinase) activity. Nine sedentary men were studied before and after 7 days of exercise training (1 h/day, ≈75% maximal oxygen consumption). Insulin sensitivity was determined with a euglycemic-hyperinsulinemic glucose clamp in the sedentary state and 15–17 h after the final exercise bout. PI 3-kinase activity was determined from samples (vastus lateralis) obtained in the fasted condition and after 60 min of submaximal insulin stimulation during the clamp. After exercise, glucose infusion rate increased ( P < 0.05) significantly (means ± SE, 7.8 ± 0.5 vs. 9.8 ± 0.8 mg ⋅ kg−1 ⋅ min−1), indicating improved insulin sensitivity. Insulin-stimulated (insulin stimulated/fasting) phosphotyrosine immunoprecipitable PI 3-kinase activity also increased significantly ( P < 0.05) with exercise (3.1 ± 0.8-fold) compared with the sedentary condition (1.3 ± 0.1-fold). There was no change in fasting PI 3-kinase activity. These data suggest that an enhancement of insulin signal transduction in skeletal muscle may contribute to the improvement in insulin action with exercise.

scholarly journals NMR-Based Metabolomic Analysis and Microbial Composition of Soil Supporting Burkea africana Growth

Metabolites ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 402
Author(s):  
Lufuno Ethel Nemadodzi ◽  
Jacques Vervoort ◽  
Gerhard Prinsloo
Keyword(s):  
Mass Spectrometry Analysis ◽  
Metabolomic Analysis ◽  
Microbial Composition ◽  
Soil Microbial Diversity ◽  
Penicillium Species ◽  
Bacterial Composition ◽  
Spectrometry Analysis ◽  
Soil Microbial ◽  
Natural Establishment ◽  
Survival Of Seedlings

Burkea africana is a leguminous tree used for medicinal purposes, growing in clusters, on soils impoverished from most nutrients. The study aimed to determine the factors responsible for successful reproduction and establishment of the B. africana trees in nature, as all efforts for commercial production has been proven unsuccessful. An investigation was carried out to determine the metabolomic profile, chemical composition, and microbial composition of the soils where B. africana grows (Burkea soil) versus the soil where it does not grow (non-Burkea soil). 1H-NMR metabolomic analysis showed different metabolites in the respective soils. Trehalose and betaine, as well as a choline-like and carnitine-like compound, were found to be in higher concentration in Burkea soils, whereas, acetate, lactate, and formate were concentrated in non-Burkea soils. Liquid Chromatography-Mass Spectrometry analysis revealed the presence of numerous amino acids such as aspartic acid and glutamine to be higher in Burkea soils. Since it was previously suggested that the soil microbial diversity is the major driver for establishment and survival of seedlings in nature, Deoxyribonucleic acid (DNA) was extracted and a BLAST analysis conducted for species identification. Penicillium species was found to be highly prevalent and discriminant between the two soils, associated with the Burkea soils. No differences in the bacterial composition of Burkea and non-Burkea soils were observed. The variances in fungal composition suggests that species supremacy play a role in development of B. africana trees and is responsible for creating a supporting environment for natural establishment and survival of seedlings.

scholarly journals Preferent Diaphragmatic Involvement in TK2 Deficiency: An Autopsy Case Study

2021 ◽  
Vol 22 (11) ◽  
pp. 5598
Author(s):  
Sara Laine-Menéndez ◽  
Cristina Domínguez-González ◽  
Alberto Blázquez ◽  
Aitor Delmiro ◽  
Inés García-Consuegra ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Late Onset ◽  
Protein Profile ◽  
Mass Spectrometry Analysis ◽  
Fructose Bisphosphate ◽  
Catalytic Subunits ◽  
Peroxiredoxin 6 ◽  
Spectrometry Analysis ◽  
Fructose Bisphosphate Aldolase

Our goal was to analyze postmortem tissues of an adult patient with late-onset thymidine kinase 2 (TK2) deficiency who died of respiratory failure. Compared with control tissues, we found a low mtDNA content in the patient’s skeletal muscle, liver, kidney, small intestine, and particularly in the diaphragm, whereas heart and brain tissue showed normal mtDNA levels. mtDNA deletions were present in skeletal muscle and diaphragm. All tissues showed a low content of OXPHOS subunits, and this was especially evident in diaphragm, which also exhibited an abnormal protein profile, expression of non-muscular β-actin and loss of GAPDH and α-actin. MALDI-TOF/TOF mass spectrometry analysis demonstrated the loss of the enzyme fructose-bisphosphate aldolase, and enrichment for serum albumin in the patient’s diaphragm tissue. The TK2-deficient patient’s diaphragm showed a more profound loss of OXPHOS proteins, with lower levels of catalase, peroxiredoxin 6, cytosolic superoxide dismutase, p62 and the catalytic subunits of proteasome than diaphragms of ventilated controls. Strong overexpression of TK1 was observed in all tissues of the patient with diaphragm showing the highest levels. TK2 deficiency induces a more profound dysfunction of the diaphragm than of other tissues, which manifests as loss of OXPHOS and glycolytic proteins, sarcomeric components, antioxidants and overactivation of the TK1 salvage pathway that is not attributed to mechanical ventilation.

scholarly journals Transcriptome Analysis of Agmatine and Putrescine Catabolism in Pseudomonas aeruginosa PAO1

2008 ◽  
Vol 190 (6) ◽  
pp. 1966-1975 ◽  
Author(s):  
Han Ting Chou ◽  
Dong-Hyeon Kwon ◽  
Mohamed Hegazy ◽  
Chung-Dar Lu
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Wide Spectrum ◽  
Tricarboxylic Acid Cycle ◽  
Mass Spectrometry Analysis ◽  
Growth Defect ◽  
Succinic Semialdehyde ◽  
Succinic Semialdehyde Dehydrogenase ◽  
Spectrometry Analysis ◽  
Semialdehyde Dehydrogenase

ABSTRACT Polyamines (putrescine, spermidine, and spermine) are major organic polycations essential for a wide spectrum of cellular processes. The cells require mechanisms to maintain homeostasis of intracellular polyamines to prevent otherwise severe adverse effects. We performed a detailed transcriptome profile analysis of Pseudomonas aeruginosa in response to agmatine and putrescine with an emphasis in polyamine catabolism. Agmatine serves as the precursor compound for putrescine (and hence spermidine and spermine), which was proposed to convert into 4-aminobutyrate (GABA) and succinate before entering the tricarboxylic acid cycle in support of cell growth, as the sole source of carbon and nitrogen. Two acetylpolyamine amidohydrolases, AphA and AphB, were found to be involved in the conversion of agmatine into putrescine. Enzymatic products of AphA were confirmed by mass spectrometry analysis. Interestingly, the alanine-pyruvate cycle was shown to be indispensable for polyamine utilization. The newly identified dadRAX locus encoding the regulator alanine transaminase and racemase coupled with SpuC, the major putrescine-pyruvate transaminase, were key components to maintaining alanine homeostasis. Corresponding mutant strains were severely hampered in polyamine utilization. On the other hand, an alternative γ-glutamylation pathway for the conversion of putrescine into GABA is present in some organisms. Subsequently, GabD, GabT, and PA5313 were identified for GABA utilization. The growth defect of the PA5313 gabT double mutant in GABA suggested the importance of these two transaminases. The succinic-semialdehyde dehydrogenase activity of GabD and its induction by GABA were also demonstrated in vitro. Polyamine utilization in general was proven to be independent of the PhoPQ two-component system, even though a modest induction of this operon was induced by polyamines. Multiple potent catabolic pathways, as depicted in this study, could serve pivotal roles in the control of intracellular polyamine levels.

Creatine kinase isoenzyme MM variants in skeletal muscle and plasma from marathon runners.

1986 ◽  
Vol 32 (1) ◽  
pp. 41-44 ◽  
Author(s):  
F S Apple ◽  
M A Rogers ◽  
J L Ivy
Keyword(s):  
Skeletal Muscle ◽  
Creatine Kinase ◽  
Strenuous Exercise ◽  
Variant Form ◽  
Distance Runners ◽  
Long Distance ◽  
Marathon Runners ◽  
Creatine Kinase Isoenzyme ◽  
Before And After

Abstract We investigated the patterns of variants of creatine kinase isoenzyme MM (CK-MM) in gastrocnemius muscle and plasma sampled from male and female long-distance runners before and after a marathon race. The proportions of CK-MM variants MM1 (pI 6.90) and MM2 (pI 6.62), identified in the skeletal muscle from both sexes, did not differ significantly from those in skeletal muscle from nonrunning controls or from heart muscle. CK-MM1 was the major (84-85% of total CK-MM) variant form. Patterns of CK-MM in plasma collected from male runners 24, 48, 72, and 96 h after the race were similar to those for female runners, but we detected two new additional variants, which we designate MM1B (pI 6.76) and MM2B (pI 6.49). For both sexes the total CK-MM activities in plasma were significantly (p less than 0.05) greater after the race, but the women's total CK-MM activities were significantly (p less than 0.05) less than the men's. The rates of disappearance of MM1, MM2, and MM3 from plasma after the race differed significantly (p less than 0.05) between men and women, MM1 clearing the fastest. Determination of the CK-MM variants in plasma after strenuous exercise may be of help in assessing CK release from injured skeletal muscle.

Epinephrine increases tricarboxylic acid cycle intermediates in human skeletal muscle

1991 ◽  
Vol 260 (3) ◽  
pp. E436-E439 ◽  
Author(s):  
M. K. Spencer ◽  
A. Katz ◽  
I. Raz
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Adenine Nucleotides ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acid Cycle ◽  
Quadriceps Femoris Muscle ◽  
Before And After ◽  
Tricarboxylic Acid Cycle Intermediates ◽  
Femoris Muscle

The effects of epinephrine (E) and insulin infusions on the contents of tricarboxylic acid cycle intermediates (TCAI), adenine nucleotides and their catabolites, and amino acids in skeletal muscle have been investigated. Eight men were studied on two separate occasions: 1) during 120 min of euglycemic hyperinsulinemia (UH, approximately 5 mM; 40 mU.m-2.min-1) and 2) during UH while E was infused (UHE, 0.05 microgram.kg-1.min-1). Biopsies were taken from the quadriceps femoris muscle before and after each clamp. The sum of citrate, malate, and fumarate in muscle did not change significantly during UH (P greater than 0.05) but doubled during UHE (P less than 0.001). There were no significant changes in any of the adenine nucleotides, their catabolites (including inosine monophosphate), or aspartate during UH and UHE (P greater than 0.05); nor were there any significant changes in pyruvate or alanine contents during UH (P greater than 0.05). On the other hand, there were significant increases in pyruvate and alanine contents during UHE (P less than 0.01 and 0.05, respectively), suggesting that there was increased production of 2-oxoglutarate (a TCAI) via the alanine aminotransferase (ALT) reaction. It is concluded that E infusion increases the contents of TCAI in human skeletal muscle, and it is likely that at least part of the increase is attributable to increased flux through the ALT reaction.

scholarly journals Multi-element effects on arsenate accumulation in a geochemical matrix determined using µ-XRF, µ-XANES and spatial statistics

2019 ◽  
Vol 26 (6) ◽  
pp. 1967-1979 ◽  
Author(s):  
Aakriti Sharma ◽  
Amanda Muyskens ◽  
Joseph Guinness ◽  
Matthew L. Polizzotto ◽  
Montserrat Fuentes ◽  
...  
Keyword(s):  
Secondary Ion Mass Spectrometry ◽  
Spatial Regression ◽  
Model Systems ◽  
Mass Spectrometry Analysis ◽  
Soil Matrix ◽  
Edge Structure ◽  
X Ray ◽  
Spectrometry Analysis ◽  
Before And After ◽  
Partial Correlations

Soils regulate the environmental impacts of trace elements, but direct measurements of reaction mechanisms in these complex, multi-component systems can be challenging. The objective of this work was to develop approaches for assessing effects of co-localized geochemical matrix elements on the accumulation and chemical speciation of arsenate applied to a soil matrix. Synchrotron X-ray fluorescence microprobe (µ-XRF) images collected across 100 µm × 100 µm and 10 µm × 10 µm regions of a naturally weathered soil sand-grain coating before and after treatment with As(V) solution showed strong positive partial correlations (r′ = 0.77 and 0.64, respectively) between accumulated As and soil Fe, with weaker partial correlations (r′ > 0.1) between As and Ca, and As and Zn in the larger image. Spatial and non-spatial regression models revealed a dominant contribution of Fe and minor contributions of Ca and Ti in predicting accumulated As, depending on the size of the sample area analyzed. Time-of-flight secondary ion mass spectrometry analysis of an area of the sand grain showed a significant correlation (r = 0.51) between Fe and Al, so effects of Fe versus Al (hydr)oxides on accumulated As could not be separated. Fitting results from 25 As K-edge microscale X-ray absorption near-edge structure (µ-XANES) spectra collected across a separate 10 µm × 10 µm region showed ∼60% variation in proportions of Fe(III) and Al(III)-bound As(V) standards, and fits to µ-XANES spectra collected across the 100 µm × 100 µm region were more variable. Consistent with insights from studies on model systems, the results obtained here indicate a dominance of Fe and possibly Al (hydr)oxides in controlling As(V) accumulation within microsites of the soil matrix analyzed, but the analyses inferred minor augmentation from co-localized Ti, Ca and possibly Zn.

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