31P NMR quantitation of phosphorus metabolites in rat heart and skeletal muscle in vivo

2001 ◽  
Vol 281 (2) ◽  
pp. H882-H887 ◽  
Author(s):  
Sam Hitchins ◽  
Julie M. Cieslar ◽  
Geoffrey P. Dobson
Keyword(s):  
Skeletal Muscle ◽  
Rat Heart ◽  
Signal Amplitude ◽  
Phosphorus Compounds ◽  
External Reference ◽  
Significant Difference ◽  
External Standard ◽  
Phosphorus Metabolites ◽  
Tissue Phosphorus

The aim of this study was to examine two methods of 31P NMR quantitation of phosphocreatine (PCr), ATP, and Pi in rat heart and skeletal muscle in vivo. The first method employed an external standard of phenylphosphonic acid (PPA; 10 mM), and the second method used an enzymatic measurement of tissue ATP equated to the area under the βATP peak. With the use of the external standard, the concentrations of ATP, PCr, and Pi in the rat heart were 4.48 ± 0.33, 9.21 ± 0.65, and 2.25 ± 0.16 μmol/g wet wt, respectively. With the use of the internal ATP standard, measured on the same tissue, the contents (means ± SE) were 4.78 ± 0.19, 9.83 ± 0.18, and 2.51 ± 0.33 μmol/g wet wt, respectively ( n = 7). In skeletal muscle, ATP, PCr, and Pi were 6.09 ± 0.19, 23.44 ± 0.88, and 1.81 ± 0.18 μmol/g wet wt using the PPA standard and 6.03 ± 0.19, 23.30 ± 1.30, and 1.82 ± 0.19 μmol/g wet wt using the internal ATP standard ( n = 6). There was no significant difference for each metabolite as measured by the two methods of quantification in heart or skeletal muscle. The results validate the use of an external reference positioned symmetrically above the coil and imply that each has similar NMR sensitivities (similar signal amplitude per mole of 31P between PPA and tissue phosphorus compounds). We conclude that PCr, ATP, and Pi are nearly 100% visible in the normoxic heart and nonworking skeletal muscle given the errors of measurement.

Tissue spaces in rat heart, liver, and skeletal muscle in vivo

1998 ◽  
Vol 275 (5) ◽  
pp. R1530-R1536 ◽  
Author(s):  
Julie Cieslar ◽  
Ming-Ta Huang ◽  
Geoffrey P. Dobson
Keyword(s):  
Skeletal Muscle ◽  
Rat Heart ◽  
Gastrocnemius Muscle ◽  
Interstitial Space ◽  
Glyceric Acid ◽  
Sprague Dawley ◽  
Tissue Water ◽  
Intracellular Space ◽  
Total Tissue

Tissue spaces were determined in rat heart, liver, and skeletal muscle in vivo using isotopically labeled [14C]inulin. Tracer was injected into the jugular vein of pentobarbital-anesthetized male Sprague-Dawley rats. After a 30-min equilibration period, a blood sample was taken, and heart, liver, and gastrocnemius muscle were excised and immediately freeze clamped at liquid nitrogen temperatures. The extracellular inulin space was 0.209 ± 0.006 ( n = 13), 0.203 ± 0.080 ( n = 7), and 0.124 ± 0.006 (SE) ml/g wet wt tissue ( n = 8) for heart, liver, and skeletal muscle, respectively. Total tissue water was 0.791 ± 0.005 ( n = 9), 0.732 ± 0.002 ( n = 9), and 0.755 ± 0.005 ml/g wet wt tissue ( n = 10) for heart, liver, and skeletal muscle, respectively. Expressed as a percentage of total tissue water, the intracellular space was 73.6, 72.2, and 83.7% for heart, liver, and skeletal muscle, respectively. With use of 2,3-diphospho-d-glyceric acid as a vascular marker, the interstitial space was calculated by subtracting the counts in tissue due to whole blood from total tissue counts and dividing by plasma counts. The interstitial space was 18.8, 22.4, and 14.5% of total tissue water, with accompanying plasma spaces of 7.7, 5.3, and 1.8% for heart, liver, and gastrocnemius muscle, respectively. The tracer method used in this study provides a quantitative assessment of water distribution in tissues of nonnephrectomized rats that has applications for calculation of tissue ion and metabolite concentrations, gradients, and fluxes under normal and pathophysiological conditions.

Steroid effects on permeability of rat thymic lymphocytes to α-aminoisobutyrate

1963 ◽  
Vol 205 (3) ◽  
pp. 446-452 ◽  
Author(s):  
Melvin Blecher
Keyword(s):  
Skeletal Muscle ◽  
Extracellular Fluid ◽  
In Vivo Studies ◽  
Metabolic Inhibitors ◽  
Active Process ◽  
Low Concentrations ◽  
Significant Difference ◽  
Adrenalectomized Rats

In vitro studies of the flux of α-aminoisobutyrate-1-C14 (AIB) between rat thymic lymphocytes and extracellular fluid have revealed that: a) the amino acid enters cells but is not further metabolized; b) at low concentrations, similar to those of amino acids in plasma, the net influx and efflux of AIB exhibit properties of an active process; and c) influx of AIB is inhibited, and efflux stimulated, by deoxycorticosterone (DOC), by metabolic inhibitors, and by other specific steroids. In vivo studies of the distribution of AIB between serum and tissue demonstrated that administration of DOC to adrenalectomized rats inhibited concentration of AIB by thymus, diaphragm, and skeletal muscle, augmented uptake by liver, and increased the serum level of AIB. Prior adrenalectomy of donor rats resulted in no change from normal in the in vitro capacity of thymic lymphocytes to take up AIB. There was no significant difference from normal in the in vivo concentration of AIB by thymus, liver, and skeletal muscle of adrenalectomized rats, although uptake by diaphragm was decreased compared to normal control animals.

In vivo insulin signaling through PI3-kinase is impaired in skeletal muscle of adult rat offspring exposed to ethanol in utero

2005 ◽  
Vol 99 (2) ◽  
pp. 528-534 ◽  
Author(s):  
Li Chen ◽  
Xing-Hai Yao ◽  
B. L. G. Nyomba
Keyword(s):  
Insulin Resistance ◽  
Skeletal Muscle ◽  
Insulin Receptor ◽  
Insulin Signaling ◽  
In Utero ◽  
Pi3 Kinase ◽  
Adult Rat ◽  
Rat Offspring ◽  
Significant Difference

It is now known that prenatal ethanol (EtOH) exposure is associated with impaired glucose tolerance and insulin resistance in rat offspring, but the underlying mechanism(s) is not known. To test the hypothesis that in vivo insulin signaling through phosphatidylinositol 3 (PI3)-kinase is reduced in skeletal muscle of adult rat offspring exposed to EtOH in utero, we gave insulin intravenously to these rats and probed steps in the PI3-kinase insulin signaling pathway. After insulin treatment, EtOH-exposed rats had decreased tyrosine phosphorylation of the insulin receptor β-subunit and of insulin receptor substrate-1 (IRS-1), as well as reduced IRS-1-associated PI3-kinase in the gastrocnemius muscle compared with control rats. There was no significant difference in basal or insulin-stimulated Akt activity between EtOH-exposed rats and controls. Insulin-stimulated PKC isoform ζ phosphorylation and membrane association were reduced in EtOH-exposed rats compared with controls. Muscle insulin binding and peptide contents of insulin receptor, IRS-1, p85 subunit of PI3-kinase, Akt/PKB, and atypical PKC isoform ζ were not different between EtOH-exposed rats and controls. Thus insulin resistance in rat offspring exposed to EtOH in utero may be explained, at least in part, by impaired insulin signaling through the PI3-kinase pathway in skeletal muscle.

Phase-modulated rotating-frame imaging of phosphorus metabolites in skeletal muscle in vivo

1987 ◽  
Vol 15 (5) ◽  
pp. 869-870
Author(s):  
MARTIN J. BLACKLEDGE ◽  
R. A. JOHN CHALLISS ◽  
GEORGE K. RADDA
Keyword(s):  
Skeletal Muscle ◽  
Rotating Frame ◽  
Phosphorus Metabolites

scholarly journals Production of monoclonal antibodies against Streptococcus mutans antigens

2006 ◽  
Vol 20 (4) ◽  
pp. 297-302 ◽  
Author(s):  
Antonio Carlos Victor Canettieri ◽  
Fujiko Yamasiro Kretchetoff ◽  
Cristiane Yumi Koga-Ito ◽  
Daniella Moreira ◽  
Fabio José Condino Fujarra ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Monoclonal Antibodies ◽  
Streptococcus Mutans ◽  
Cardiac Tissue ◽  
Cross Reactivity ◽  
Oral Streptococci ◽  
Significant Difference ◽  
Common Antigens ◽  
Human Cardiac Tissue

Several studies have been conducted in the last decades aiming to obtain an anti-caries vaccine, however some studies have demonstrated cross reactivity between Streptococcus mutans surface antigens and the human cardiac tissue. In this work, the reactivity of five anti-Streptococcus mutans monoclonal antibodies (MoAb) (24A, 56G, C8, E8 and F6) was tested against oral streptococci, cardiac antigens and skeletal and cardiac myosins, aiming to evaluate the specificity of these MoAb. The hybrid producers of immunoglobulins of the IgG2b class were cloned by limit dilution and expanded in vivo. MoAb were tested by ELISA. The hybrid 24A reacted with S. mutans CCT 1910, S. salivarius CCT 0365 and S. pyogenes T23. No reactivity difference was observed among the tested species. Cross reactivity with heart and cardiac myosin was not confirmed and only reaction with myosin of skeletal muscle was observed (p = 0.0381). The hybrid 56G reacted with all the tested microorganisms and there was statistically significant difference between S. mutans and S. pyogenes T23 (p < 0.001). This hybrid also reacted with myosin of skeletal muscle (p = 0.0095). C8, E8 and F6 presented low reactivity against oral streptococci strains and no reactivity against cardiac antigens. The data of this study showed that the 24A and 56G anti-S. mutans MoAb presented reactivity with S. pyogenes and S. salivarius, reinforcing the occurrence of common antigens between these species. The tested MoAb presented low cross-reactivity with myosin of skeletal muscle, but anti-heart activity could not be confirmed.

scholarly journals Sex differences in mitochondrial Ca2+ handling in mouse fast-twitch skeletal muscle in vivo

2020 ◽  
Vol 128 (2) ◽  
pp. 241-251 ◽  
Author(s):  
Daiki Watanabe ◽  
Koji Hatakeyama ◽  
Ryo Ikegami ◽  
Hiroaki Eshima ◽  
Kazuyoshi Yagishita ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Sex Differences ◽  
Cyclopiazonic Acid ◽  
Atpase Inhibitor ◽  
Fiber Volume ◽  
Male And Female ◽  
Significant Difference ◽  
Mitochondrial Volume ◽  
Fast Twitch

We investigated sex differences in mitochondrial Ca2+ handling properties in mouse fast-twitch skeletal muscle. Changes in cytoplasmic Ca2+ concentration ([Ca2+]cyto) were measured in vivo using tibialis anterior muscles from male and female mice. The muscles were exposed to increasing concentrations of cyclopiazonic acid [CPA; sarcoplasmic reticulum (SR) Ca2+-ATPase inhibitor] (from 10 to 30 to 50 μM at 10 min intervals). Thirty minutes after treatment, [Ca2+]cyto was increased by 31.6 ± 2.0% and 13.5 ± 4.5% of initial [Ca2+]cyto in male and female muscles, respectively, and there was a significant difference between sexes. However, muscle preincubation for 5 min with 10 μM carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (an inhibitor of mitochondria Ca2+ uptake) eradicated this difference between sexes with respect to the CPA-induced [Ca2+]cyto increase. Both intermyofibrillar mitochondrial number and volume, assessed in longitudinal fiber sections, were higher in females compared with males (mitochondria number: 13.1 ± 1.0 in males vs. 19.9 ± 2.3 in females; mitochondrial volume: 0.034 ± 0.004 μm3/μm3 fiber volume in males vs. 0.066 ± 0.008 μm3/μm3 fiber volume in females, both P < 0.05). There were no sex differences in the content of SR Ca2+-ATPase, mitochondrial Ca2+ uniporter, mitofusin (Mfn) 1, or Mfn2. These results suggest that 1) mitochondrial Ca2+ uptake ability is greater in female than male myocytes, and 2) this superior Ca2+ uptake ability of female myocytes is due, partly, to the higher intermyofibrillar mitochondrial content but not to the expression of mitochondrial proteins related to mitochondrial Ca2+ uptake. NEW & NOTEWORTHY This investigation presents evidence that female versus male fast-twitch muscle exhibits a greater mitochondrial calcium ion uptake capability that is partly conferred by the higher intermyofibrillar mitochondrial volume density.

scholarly journals Comparison of in vivo postexercise phosphocreatine recovery and resting ATP synthesis flux for the assessment of skeletal muscle mitochondrial function

2010 ◽  
Vol 299 (5) ◽  
pp. C1136-C1143 ◽  
Author(s):  
N. M. A. van den Broek ◽  
J. Ciapaite ◽  
K. Nicolay ◽  
J. J. Prompers
Keyword(s):  
Skeletal Muscle ◽  
Oxygen Consumption ◽  
Mitochondrial Dysfunction ◽  
Mitochondrial Function ◽  
Atp Synthesis ◽  
Resonance Spectroscopy ◽  
Isolated Mitochondria ◽  
Significant Difference

31P magnetic resonance spectroscopy (MRS) has been used to assess skeletal muscle mitochondrial function in vivo by measuring 1) phosphocreatine (PCr) recovery after exercise or 2) resting ATP synthesis flux with saturation transfer (ST). In this study, we compared both parameters in a rat model of mitochondrial dysfunction with the aim of establishing the most appropriate method for the assessment of in vivo muscle mitochondrial function. Mitochondrial dysfunction was induced in adult Wistar rats by daily subcutaneous injections with the complex I inhibitor diphenyleneiodonium (DPI) for 2 wk. In vivo 31P MRS measurements were supplemented by in vitro measurements of oxygen consumption in isolated mitochondria. Two weeks of DPI treatment induced mitochondrial dysfunction, as evidenced by a 20% lower maximal ADP-stimulated oxygen consumption rate in isolated mitochondria from DPI-treated rats oxidizing pyruvate plus malate. This was paralleled by a 46% decrease in in vivo oxidative capacity, determined from postexercise PCr recovery. Interestingly, no significant difference in resting, ST-based ATP synthesis flux was observed between DPI-treated rats and controls. These results show that PCr recovery after exercise has a more direct relationship with skeletal muscle mitochondrial function than the ATP synthesis flux measured with 31P ST MRS in the resting state.

In-vivo evaluation of the effect of cyanoacrylate on prosthetic vascular graft infection – does cyanoacrylate increase the severity of infection?

VASA ◽  
2020 ◽  
Vol 49 (4) ◽  
pp. 281-284
Author(s):  
Atıf Yolgosteren ◽  
Gencehan Kumtepe ◽  
Melda Payaslioglu ◽  
Cuneyt Ozakin
Keyword(s):  
Vascular Graft ◽  
Graft Infection ◽  
Vascular Graft Infection ◽  
Group 4 ◽  
Significant Difference ◽  
Group 3 ◽  
Group 2 ◽  
Prosthetic Vascular Graft Infection ◽  
Group 1

Summary. Background: Prosthetic vascular graft infection (PVGI) is a complication with high mortality. Cyanoacrylate (CA) is an adhesive which has been used in a number of surgical procedures. In this in-vivo study, we aimed to evaluate the relationship between PVGI and CA. Materials and methods: Thirty-two rats were equally divided into four groups. Pouch was formed on back of rats until deep fascia. In group 1, vascular graft with polyethyleneterephthalate (PET) was placed into pouch. In group 2, MRSA strain with a density of 1 ml 0.5 MacFarland was injected into pouch. In group 3, 1 cm 2 vascular graft with PET piece was placed into pouch and MRSA strain with a density of 1 ml 0.5 MacFarland was injected. In group 4, 1 cm 2 vascular graft with PET piece impregnated with N-butyl cyanoacrylate-based adhesive was placed and MRSA strain with a density of 1 ml 0.5 MacFarland was injected. All rats were scarified in 96th hour, culture samples were taken where intervention was performed and were evaluated microbiologically. Bacteria reproducing in each group were numerically evaluated based on colony-forming unit (CFU/ml) and compared by taking their average. Results: MRSA reproduction of 0 CFU/ml in group 1, of 1410 CFU/ml in group 2, of 180 200 CFU/ml in group 3 and of 625 300 CFU/ml in group 4 was present. A statistically significant difference was present between group 1 and group 4 (p < 0.01), between group 2 and group 4 (p < 0.01), between group 3 and group 4 (p < 0.05). In terms of reproduction, no statistically significant difference was found in group 1, group 2, group 3 in themselves. Conclusions: We observed that the rate of infection increased in the cyanoacyrylate group where cyanoacrylate was used. We think that surgeon should be more careful in using CA in vascular surgery.

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