scholarly journals Direct measurement of the lumped constant for 2-deoxy-[1-14C]glucose in vivo in human skeletal muscle

2000 ◽  
Vol 279 (1) ◽  
pp. E228-E233 ◽  
Author(s):  
Tapio Utriainen ◽  
Stefania Lovisatti ◽  
Sari Mäkimattila ◽  
Alessandra Bertoldo ◽  
Susan Weintraub ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Insulin Infusion ◽  
Normal Subjects ◽  
Normal Male ◽  
Wide Range ◽  
Lumped Constant ◽  
Male Subjects ◽  
Forearm Vein

The lumped constant (LC) is used to convert the clearance rate of 2-deoxy-d-glucose (2-DGcr) to that of glucose (Glccr). There are currently no data to validate the widely used assumption of an LC of 1.0 for human skeletal muscle. We determined the LC for 2-deoxy-[1-14C]glucose (2-DG) in 18 normal male subjects (age, 29 ± 2 yr; body mass index, 24.8 ± 0.8 kg/m2) after an overnight fast and during physiological (1 mU · kg−1· min−1insulin infusion for 180 min) and supraphysiological (5 mU · kg−1· min−1insulin infusion for 180 min) hyperinsulinemic conditions. Normoglycemia was maintained with the euglycemic clamp technique. The LC was measured directly with the use of a novel triple tracer-based method. [3-3H]glucose, 2-[1-14C]DG, and [12C]mannitol (Man) were injected as a bolus into the brachial artery. The concentrations of [3-3H]glucose and 2-[1-14C]DG (dpm/ml plasma) and of Man (μmol/l) were determined in 50 blood samples withdrawn from the ipsilateral deep forearm vein over 15 min after the bolus injection. The LC was calculated by a formula involving blood flow calculated from Man and the Glccrand 2-DGcr. The LC averaged 1.26 ± 0.08 (range 1.06–1.43), 1.15 ± 0.05 (0.99–1.39), and 1.18 ± 0.05 (0.97–1.37) under fasting conditions and during the 1 and 5 mU · kg−1· min−1insulin infusions (not significant between the different insulin concentrations, mean LC = 1.2, P < 0.01 vs. 1.0). We conclude that, in normal subjects, the LC for 2-DG in human skeletal muscle is constant over a wide range of insulin concentrations and averages 1.2.

Regulation of hexokinase II and glycogen synthase mRNA, protein, and activity in human muscle

1995 ◽  
Vol 269 (4) ◽  
pp. E701-E708 ◽  
Author(s):  
L. J. Mandarino ◽  
R. L. Printz ◽  
K. A. Cusi ◽  
P. Kinchington ◽  
R. M. O'Doherty ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Insulin Infusion ◽  
Glycogen Synthase ◽  
Normal Subjects ◽  
Hexokinase Ii ◽  
Glut 4 ◽  
Before And After ◽  
Muscle Biopsies

Insulin regulates the activity of key enzymes of glucose metabolism in skeletal muscle by altering transcription or translation or by producing activity-altering modifications of preexisting enzyme molecules. Because of the small size of percutaneous muscle biopsies, these phenomena have been difficult to study in humans. This study was performed to determine how physiological hyperinsulinemia regulates the activities of hexokinase (HK), glycogen synthase (GS), and GLUT-4 in human skeletal muscle in vivo. We determined mRNA abundance, protein content, and activities for these proteins in muscle biopsies before and after a hyperinsulinemic clamp in normal subjects. HK I, HK II, GS, and GLUT-4 were expressed in muscle. HK II accounted for 80% of total HK activity and was increased by insulin from a basal value of 2.11 +/- 0.26 to 3.35 +/- 0.47 pmol.min-1.mg protein-1 (P < 0.05); HK I activity was unaffected. Insulin increased GS activity from 3.85 +/- 0.82 to 6.06 +/- 0.49 nmol.min-1.mg-1 (P < 0.01). HK II mRNA was increased 3.3 +/- 1.3-fold (P < 0.05) by insulin infusion. HK I, GS, and GLUT-4 mRNA and protein were unaffected. Because insulin infusion increased HK II but not GS mRNA, we conclude that HK II and GS may be regulated by insulin by different mechanisms in human skeletal muscle.

Effect of insulin on intracellular pH and phosphate metabolism in human skeletal muscle in vivo

1991 ◽  
Vol 81 (1) ◽  
pp. 123-128 ◽  
Author(s):  
D. J. Taylor ◽  
S. W. Coppack ◽  
T. A. D. Cadoux-Hudson ◽  
G. J. Kemp ◽  
G. K. Radda ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Intracellular Ph ◽  
Inorganic Phosphate ◽  
Human Skeletal Muscle ◽  
Muscle Metabolism ◽  
Normal Subjects ◽  
Phosphate Concentration ◽  
Resonance Spectroscopy ◽  
Phosphorus Containing

1. 31P nuclear magnetic resonance spectroscopy and the hyperinsulinaemic-euglycaemic clamp were used simultaneously to assess the effect of insulin on intracellular pH and the major phosphorus-containing metabolites of normal human skeletal muscle in vivo in four normal subjects. 2. Insulin and glucose were infused for 120 min. Plasma insulin increased approximately 10-fold over pre-clamp levels (5.6 ± 0.9 m-units/l pre-clamp and 54 ± 5 m-units/l over the last hour of infusion; mean ± sem, n = 4). Plasma glucose concentration did not change significantly (5.4 ± 0.2 mmol/l pre-clamp and 5.5 ± 0.1 mmol/l over the last hour of infusion). 3. Insulin and glucose infusion resulted in a decline in the intracellular pH of forearm muscle of 0.027 ± 0.007 unit/h (P < 0.01), whereas in control studies of the same subjects, pH rose by 0.046 ± 0.005 unit/h (P < 0.001). 4. In the clamp studies, intracellular inorganic phosphate concentration rose by 18%/h, whereas ATP, phosphocreatine and phosphomonoester concentrations did not change. In plasma, inorganic phosphate concentration was 1.16 ± 0.05 mmol/l before infusion, and this decreased by a mean rate of 0.14 mmol h−1 l−1. No change was observed in any of these intracellular metabolites in the control studies. 5. The results show that, under physiological conditions, insulin does not raise intracellular pH in human muscle, and thus cannot influence muscle metabolism by this mechanism. The results also suggest that insulin causes a primary increase in the next flux of inorganic phosphate across the muscle cell membrane.

scholarly journals Novel Method for Detection of Reactive Oxygen Species In Vivo in Human Skeletal Muscle

2014 ◽  
pp. 387-392 ◽  
Author(s):  
J. D. LA FAVOR ◽  
E. J. ANDERSON ◽  
R. C. HICKNER
Keyword(s):  
Reactive Oxygen Species ◽  
Skeletal Muscle ◽  
Human Skeletal Muscle ◽  
Vastus Lateralis ◽  
Reactive Oxygen ◽  
High Reactivity ◽  
Oxygen Species ◽  
Microdialysis Probe ◽  
Wide Range

Excessive production of reactive oxygen species (ROS) are implicated in the pathogenesis of numerous disease states. However, direct measurement of in vivo ROS in humans has remained elusive due to limited access to appropriate tissue beds and the inherently short half-lives and high reactivity of ROS. Herein, we describe a novel technique by which to measure in vivo ROS in human skeletal muscle. Microdialysis probes were inserted into the vastus lateralis of eight healthy volunteers. Amplex Ultrared, a highly specific fluorogenic substrate for hydrogen peroxide (H2O2), and horseradish peroxidase (HRP), were perfused through microdialysis probes, and outflowing dialysate was collected and fluorescence was measured. Extracellular H2O2 that crossed the microdialysis membrane was measured via fluorescence of the dialysate. Superoxide dismutase (SOD) was then added to the inflowing perfusion media to convert any superoxide crossing the microdialysis membrane to H2O2 within the microdialysis probe. Fluorescence significantly increased (P=0.005) upon SOD addition. These data demonstrate the feasibility of measuring both in vivo H2O2 and superoxide in the extracellular environment of human skeletal muscle, providing a technique with a potential application to a wide range of circulatory and metabolic studies of oxidative stress.

Evaluation of 99mTc-Label led Vitamin K4 as an Agent for Testicular Imaging

1991 ◽  
Vol 30 (01) ◽  
pp. 35-39 ◽  
Author(s):  
H. S. Durak ◽  
M. Kitapgi ◽  
B. E. Caner ◽  
R. Senekowitsch ◽  
M. T. Ercan
Keyword(s):  
Soft Tissue ◽  
Room Temperature ◽  
Normal Subjects ◽  
Left Testis ◽  
Wide Range ◽  
Activity Ratios ◽  
The Right ◽  
Radioactivity Levels

Vitamin K4 was labelled with 99mTc with an efficiency higher than 97%. The compound was stable up to 24 h at room temperature, and its biodistribution in NMRI mice indicated its in vivo stability. Blood radioactivity levels were high over a wide range. 10% of the injected activity remained in blood after 24 h. Excretion was mostly via kidneys. Only the liver and kidneys concentrated appreciable amounts of radioactivity. Testis/soft tissue ratios were 1.4 and 1.57 at 6 and 24 h, respectively. Testis/blood ratios were lower than 1. In vitro studies with mouse blood indicated that 33.9 ±9.6% of the radioactivity was associated with RBCs; it was washed out almost completely with saline. Protein binding was 28.7 ±6.3% as determined by TCA precipitation. Blood clearance of 99mTc-l<4 in normal subjects showed a slow decrease of radioactivity, reaching a plateau after 16 h at 20% of the injected activity. In scintigraphic images in men the testes could be well visualized. The right/left testis ratio was 1.08 ±0.13. Testis/soft tissue and testis/blood activity ratios were highest at 3 h. These ratios were higher than those obtained with pertechnetate at 20 min post injection.99mTc-l<4 appears to be a promising radiopharmaceutical for the scintigraphic visualization of testes.

scholarly journals Autophagy Deficiency by Atg4B Loss Leads to Metabolomic Alterations in Mice

Metabolites ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 481
Author(s):  
Gemma G. Martínez-García ◽  
Raúl F. Pérez ◽  
Álvaro F. Fernández ◽  
Sylvere Durand ◽  
Guido Kroemer ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Amino Acids ◽  
Mammalian Cells ◽  
Tissue Homeostasis ◽  
In Vivo Studies ◽  
Protective Mechanism ◽  
Cardiac Damage ◽  
Wide Range ◽  
First Time

Autophagy is an essential protective mechanism that allows mammalian cells to cope with a variety of stressors and contributes to maintaining cellular and tissue homeostasis. Due to these crucial roles and also to the fact that autophagy malfunction has been described in a wide range of pathologies, an increasing number of in vivo studies involving animal models targeting autophagy genes have been developed. In mammals, total autophagy inactivation is lethal, and constitutive knockout models lacking effectors of this route are not viable, which has hindered so far the analysis of the consequences of a systemic autophagy decline. Here, we take advantage of atg4b−/− mice, an autophagy-deficient model with only partial disruption of the process, to assess the effects of systemic reduction of autophagy on the metabolome. We describe for the first time the metabolic footprint of systemic autophagy decline, showing that impaired autophagy results in highly tissue-dependent alterations that are more accentuated in the skeletal muscle and plasma. These changes, which include changes in the levels of amino-acids, lipids, or nucleosides, sometimes resemble those that are frequently described in conditions like aging, obesity, or cardiac damage. We also discuss different hypotheses on how impaired autophagy may affect the metabolism of several tissues in mammals.

Fatty Acid Oxidation in Mitochondria from Needle Biopsy Samples of Human Skeletal Muscle

1984 ◽  
Vol 66 (2) ◽  
pp. 173-178 ◽  
Author(s):  
K. Gohil ◽  
D. A. Jones ◽  
R. H. T. Edwards
Keyword(s):  
Skeletal Muscle ◽  
Cytochrome C ◽  
Needle Biopsy ◽  
Human Skeletal Muscle ◽  
Normal Subjects ◽  
Normal Activity ◽  
Carnitine Palmitoyltransferase ◽  
Acid Oxidation ◽  
Palmitoyl Carnitine ◽  
Mitochondrial Abnormality

1. Activities for the oxidation of palmitoyl-carnitine, of palmitoyl-CoA and of carnitine palmitoyltransferase were measured in mitochondria prepared from needle biopsy samples of human skeletal muscle. Results are presented for nine normal subjects and 18 patients in whom there was evidence of mitochondrial abnormality. 2. Palmitoylcarnitine and palmitoyl-CoA oxidation were measured spectrophotometrically by following the reduction of added cytochrome c in the presence of cyanide. 3. Because of large variations in the activities between subjects it was essential to express the three activities per unit of cytochrome c oxidase activity to demonstrate unambiguous specific alterations in the activities. 4. In most of the patients the order of the three activities was similar to that in the normal subjects. However, in five cases the activity for palmitoylcarnitine oxidation was less than 4% of the mean normal value. In two of these patients, the low activity could be accounted for by very low (<10% normal) activity of carnitine palmitoyltransferase (CPT). In another two patients the activity of CPT was normal but that of palmitoyl-CoA dehydrogenase (a measure of β-oxidation) was very low.

Physical exercise increases autophagic signaling through ULK1 in human skeletal muscle

2015 ◽  
Vol 118 (8) ◽  
pp. 971-979 ◽  
Author(s):  
Andreas Buch Møller ◽  
Mikkel Holm Vendelbo ◽  
Britt Christensen ◽  
Berthil Forrest Clasen ◽  
Ann Mosegaard Bak ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Physical Exercise ◽  
Light Chain ◽  
Human Skeletal Muscle ◽  
Transgenic Animal ◽  
Dependent Manner ◽  
Healthy Humans ◽  
Transgenic Animal Models ◽  
Exercise Induced

Data from transgenic animal models suggest that exercise-induced autophagy is critical for adaptation to physical training, and that Unc-51 like kinase-1 (ULK1) serves as an important regulator of autophagy. Phosphorylation of ULK1 at Ser555 stimulates autophagy, whereas phosphorylation at Ser757 is inhibitory. To determine whether exercise regulates ULK1 phosphorylation in humans in vivo in a nutrient-dependent manner, we examined skeletal muscle biopsies from healthy humans after 1-h cycling exercise at 50% maximal O2 uptake on two occasions: 1) during a 36-h fast, and 2) during continuous glucose infusion at 0.2 kg/h. Physical exercise increased ULK1 phosphorylation at Ser555 and decreased lipidation of light chain 3B. ULK1 phosphorylation at Ser555 correlated positively with AMP-activated protein kinase-α Thr172 phosphorylation and negatively with light chain 3B lipidation. ULK1 phosphorylation at Ser757 was not affected by exercise. Fasting increased ULK1 and p62 protein expression, but did not affect exercise-induced ULK1 phosphorylation. These data demonstrate that autophagy signaling is activated in human skeletal muscle after 60 min of exercise, independently of nutritional status, and suggest that initiation of autophagy constitutes an important physiological response to exercise in humans.

In vivo quantification of pulmonary beta-adrenoceptor density in humans with (S)-[11C]CGP-12177 and PET

1993 ◽  
Vol 75 (2) ◽  
pp. 559-565 ◽  
Author(s):  
J. Ueki ◽  
C. G. Rhodes ◽  
J. M. Hughes ◽  
R. De Silva ◽  
D. C. Lefroy ◽  
...  
Keyword(s):  
Hypertrophic Cardiomyopathy ◽  
Specific Activity ◽  
Regional Distribution ◽  
High Specific Activity ◽  
Normal Subjects ◽  
Normal Male ◽  
Tissue Density ◽  
Beta Adrenoceptor ◽  
Cgp 12177

The in vivo regional distribution of pulmonary beta-adrenoceptors was imaged and quantified in humans with the hydrophilic beta-adrenoceptor antagonist (S)-CGP-12177 labeled with carbon-11 [(S)-[11C]CGP-12177] and positron emission tomography (PET). Six normal male volunteers and eight patients with hypertrophic cardiomyopathy were studied. PET scanning consisted of transmission (tissue density), C15O (blood volume), and (S)-[11C]CGP-12177 (beta-adrenoceptor) emission scans. High-specific-activity (S)-[11C]-CGP-12177 (7.1 +/- 2.0 micrograms, 6.5 +/- 2.1 GBq/mumol) was given intravenously followed by a low-specific-activity (S)-[11C]CGP-12177 injection (34.0 +/- 4.8 micrograms, 2.3 +/- 0.8 GBq/mumol). Binding capacity (Bmax) was calculated in each region of interest as picomoles per gram by normalizing it to the local extravascular tissue density. In normal subjects, average Bmax for all regions of interest was 14.8 +/- 1.6 (SD) pmol/g, which is similar to previously reported in vitro values. In both groups there were no differences in beta-adrenoceptor density between peripheral and central regions nor between right and left lungs. In patients with hypertrophic cardiomyopathy, extravascular tissue density was 24% higher than in normal subjects; Bmax per milliliter thoracic volume was correspondingly higher but was not different from that in normal subjects when expressed per gram tissue (15.8 +/- 2.6 pmol/g). These data suggest that in vivo beta-adrenoceptor density may be quantifiable in humans with the use of PET. This should offer a means to study physiological regulation through repeat measurements.

Pro- and macroglycogenolysis: relationship with exercise intensity and duration

2001 ◽  
Vol 90 (3) ◽  
pp. 873-879 ◽  
Author(s):  
T. E. Graham ◽  
K. B. Adamo ◽  
J. Shearer ◽  
I. Marchand ◽  
B. Saltin
Keyword(s):  
Skeletal Muscle ◽  
Metabolic Regulation ◽  
Human Skeletal Muscle ◽  
Exercise Period ◽  
Group 3 ◽  
Group 2 ◽  
Male Subjects ◽  
Muscle Biopsies ◽  
Over Time ◽  
Group 1

We examined the net catabolism of two pools of glycogen, proglycogen (PG) and macroglycogen (MG), in human skeletal muscle during exercise. Male subjects ( n = 21) were assigned to one of three groups. Group 1 exercised 45 min at 70% maximal O2 uptake (V˙o 2 max) and had muscle biopsies at rest, 15 min, and 45 min. Group 2 exercised at 85%V˙o 2 max to exhaustion (45.4 ± 3.4 min) and had biopsies at rest, 10 min, and exhaustion. Group 3 performed three 3-min bouts of exercise at 100%V˙o 2 max separated by 6 min of rest. Biopsies were taken at rest and after each bout. Group 1 had small MG and PG net glycogenolysis rates (ranging from 3.8 ± 1.0 to 2.4 ± 0.6 mmol glucosyl units · kg−1 · min−1) that did not change over time. In group 2, the MG glycogenolysis rate remained low and unchanged over time, whereas the PG rate was initially elevated (11.3 ± 2.3 mmol glucosyl units · kg−1 · min−1) and declined ( P ≤ 0.05) with time. During the first 10 min, PG concentration ([PG]) declined ( P ≤ 0.05), whereas MG concentration ([MG]) did not. Similarly, in group 3, in both the first and the second bouts of exercise [PG] declined ( P ≤ 0.05) and [MG] did not, although by the end of the second exercise period the [MG] was lower ( P ≤ 0.05) than the rest level. The net catabolic rates for PG in the first two exercises were 22.6 ± 6.8 and 21.8 ± 8.2 mmol glucosyl units · kg−1 · min−1, whereas the corresponding values for MG were 17.6 ± 6.0 and 10.8 ± 5.6. The MG pool appeared to be more resistant to mobilization, and, when activated, its catabolism was inhibited more rapidly than that of PG. This suggests that the metabolic regulation of the two pools must be different.

scholarly journals Comparison of the Effects of Intravenous Papaverine Hydrochloride and Oral Pavabid HP Capsulets on Regional Cerebral Blood Flow in Normal Individuals

1983 ◽  
Vol 3 (4) ◽  
pp. 442-447 ◽  
Author(s):  
Lawrence C. McHenry ◽  
David A. Stump ◽  
George Howard ◽  
Thomas T. Novack ◽  
Don H. Bivins ◽  
...  
Keyword(s):  
Blood Flow ◽  
Cerebral Blood Flow ◽  
Regional Cerebral Blood Flow ◽  
Normal Subjects ◽  
Normal Male ◽  
Regional Cerebral Blood ◽  
Normal Individuals ◽  
Left And Right ◽  
Male Subjects ◽  
Papaverine Hydrochloride

A single-blind study was conducted in 13 right-handed normal male subjects to compare the effects of oral and i.v. papaverine on regional cerebral blood flow (rCBF). Six xenon-133 inhalation rCBF measurements were performed on each subject; three tests—baseline, placebo, and drug evaluations—were carried out on each of two separate days. The oral and i.v. drugs were randomized for first-day administration. rCBF, measured as flow gray (FG), increased significantly (p ≤ 0.001) from baseline with both drug forms. Increases of 10.53% and 13.94% (left and right hemispheres, respectively) were demonstrated 90 min after a single 600-mg dose of oral papaverine. Increases of 5.09% and 8.69%, respectively, were recorded immediately after a single 100-mg dose of i. v. papaverine. FG also increased significantly (p ≤ 0.001) for both drug forms when compared to that of placebo. Placebo produced only a slight increase (not significant) with both the oral and i.v. groups. The data show that both oral and i.v. papaverine are equally effective in increasing rCBF in normal subjects.

Sign in / Sign up

Export Citation Format

Share Document