Basal, but not overload-induced, myonuclear addition is attenuated by NG-nitro-l-arginine methyl ester (l-NAME) administration

2007 ◽  
Vol 85 (6) ◽  
pp. 646-651 ◽  
Author(s):  
Scott E. Gordon ◽  
Christopher M. Westerkamp ◽  
Kathleen J. Savage ◽  
Robert C. Hickner ◽  
Sarah C. George ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Methyl Ester ◽  
Protein Content ◽  
Body Mass ◽  
Total Protein ◽  
Total Protein Content ◽  
Sham Operation ◽  
Sprague Dawley ◽  
Surgical Ablation ◽  
Cross Sectional

The purpose of this study was to examine the effect of blocking nitric oxide synthase (NOS) activity via NG-nitro-l-arginine methyl ester (l-NAME) on myonuclear addition in skeletal muscle under basal and overloaded conditions. Female Sprague–Dawley rats (approx. 220 g) were placed into 1 of the following 4 groups (n = 7–9/group): 7-day skeletal muscle overload (O), sham operation (S), skeletal muscle overload with l-NAME treatment (OLN), and sham operation with l-NAME treatment (SLN). Plantaris muscles were overloaded via bilateral surgical ablation of the gastrocnemius muscles and l-NAME (0.75 mg/mL) was administered in the animals’ daily drinking water starting 2 days prior to surgery and continued until sacrifice. Myonuclear addition was assessed as subsarcolemmal incorporation of nuclei labeled with 5-bromo-2′-deoxyuridine (approx. 25 mg·(kg body mass)–1·day–1) delivered via osmotic pump during the overload period. As expected, muscle wet mass, total protein content, fiber cross-sectional area, and myonuclear addition were significantly higher (p ≤ 0.05) in O vs. S; however, only the increase in wet mass and total protein content (per body mass) were attenuated by l-NAME administration. Interestingly, l-NAME significantly reduced myonuclear addition by 75% in nonoverloaded muscles (SLN vs. S). Muscle hepatocyte growth factor protein content increased with overload, but was unaffected by l-NAME in either loading state. These data indicate that NOS inhibition in rat plantaris muscle attenuates myonuclear addition under basal, but not overloaded, conditions.

scholarly journals Accessory lymph sacs and body fluid partitioning in the lizard, Sauromalus hispidus

1986 ◽  
Vol 121 (1) ◽  
pp. 165-177
Author(s):  
A. W. Smits
Keyword(s):  
Protein Content ◽  
Body Mass ◽  
Total Protein ◽  
Body Fluid ◽  
Extracellular Fluid ◽  
Body Fluids ◽  
Body Water ◽  
Extracellular Fluid Volume ◽  
Total Protein Content ◽  
Fluid Shifts

Chuckwalla lizards (genus Sauromalus) may accumulate substantial quantities of body fluid in extracoelomic, lateral abdominal spaces called accessory lymph sacs. The lymph sac fluid (LSF) of S. hispidus is similar to that of serum in Na+, K+ and Cl- concentrations, but the total protein content (3.58 +/− 0.20 g dl-1) is only half that measured in serum (7.05 +/− 0.26 g dl-1). These analyses confirm that LSF is an extravascular form of extracellular fluid, similar in composition to true lymph. Measurements of body fluid partitioning by dilution analysis indicate that Sauromalus hispidus Stejnejer possesses a comparatively large (38.9% body mass) and labile extracellular fluid volume (ECFV), and that the volume of LSF is dependent on the ECFV. Expansion of the ECFV (and subsequent accumulation of LSF) is observed following large, intercompartmental fluid shifts from intracellular to extracellular locations when lizards are kept inactive in simulated hibernation, are injected with KCl in amounts similar to those found in their field diet, and are hydrated with NaCl that is isotonic to their body fluids. These data collectively suggest that the lymph sacs of chuckwallas facilitate expansion of the ECFV, and may be adaptive not only as a means to store body water, but to accommodate transient shifts in body fluid from intracellular to extracellular locations.

Comparative analysis of protein content in rat mesenteric tissue, peritoneal fluid, and plasma

1990 ◽  
Vol 258 (5) ◽  
pp. G714-G718 ◽  
Author(s):  
B. J. Barber ◽  
T. J. Schultz ◽  
D. L. Randlett
Keyword(s):  
Protein Content ◽  
Total Protein ◽  
Peritoneal Fluid ◽  
Protein Concentration ◽  
Total Protein Content ◽  
Sprague Dawley ◽  
Tissue Water ◽  
Tissue Samples ◽  
The Matrix ◽  
Lowry Assay

Albumin, transferrin, and total protein concentrations were measured in the mesenteric tissue, peritoneal fluid, and plasma of 12 ketamine-Nembutal-anesthetized Sprague-Dawley rats. Tissue samples were obtained with an 8-mm trephine; tissue water content was determined by a microgravimetric method to be 5.2 +/- 0.3 microgram water/microgram dry wt. Peritoneal fluid was collected by capillary action in hematocrit tubes, and blood samples were taken from a femoral artery catheter. Total protein concentrations of plasma (5.8 +/- 0.3 g/dl) and peritoneal fluid (2.6 +/- 0.1 g/dl) were determined by Lowry assay. Ratios of peritoneal fluid and tissue densitogram areas to plasma area were used to calculate total protein content of peritoneal fluid (2.5 +/- 0.1 g/dl) and tissue (1.8 +/- 0.2 g/dl). Albumin concentrations were 1.1 +/- 0.1 g/dl for tissue, 1.4 +/- 0.1 g/dl for peritoneal fluid, and 2.8 +/- 0.1 g/dl for plasma. Transferrin concentrations were 0.09 +/- 0.01 g/dl for tissue, 0.13 +/- 0.01 g/dl for peritoneal fluid, and 0.28 +/- 0.01 g/dl for plasma. Peritoneal fluid protein concentrations were similar to values found for lymph in previous studies. Protein concentration in the tissue buttons was significantly less than that of peritoneal fluid. This contradicts the widely held assumption that the protein concentration of fluid outside the matrix is representative of a well-mixed interstitial matrix fluid protein concentration.

Whole-body composition of Xenopus laevis larvae: implications for lean body mass during development.

1998 ◽  
Vol 201 (7) ◽  
pp. 1013-1022
Author(s):  
P R Territo ◽  
A W Smits
Keyword(s):  
Body Composition ◽  
Xenopus Laevis ◽  
Protein Content ◽  
Lean Body Mass ◽  
Body Mass ◽  
Total Protein ◽  
Whole Body ◽  
Nucleic Acid Content ◽  
Total Protein Content ◽  
Physiological Measures

Body composition in developing animals has been extensively investigated in fish larvae and bird embryos. However, no studies to date have attempted to determine whole-animal body composition or lean body mass (MLB) in developing amphibians. The present study investigates how body composition changes during development in Xenopus laevis and the potential implications of MLB for substrate turnover, energy stores, oxygen consumption and other physiological measures. Whole-animal composition was determined during development from eggs (NF stage 1) to 2 weeks post-feeding (NF 50-51), which represents two-thirds of the developmental period. Wet and dry masses were found to be highly correlated, with water content remaining constant at 93 % of wet mass. Whole-animal nucleic acid content was linearly correlated with both wet and dry masses, and declined relative to mass as development progressed. Similarly, total protein content was linearly correlated with wet and dry masses; however, total protein content increased with developmental stage. Amounts of individual neutral lipids were variable although, overall, total neutral lipid content declined progressively with development. The stoichiometric energy balance paralleled the changes seen in mass-specific .MO2, with the energy primarily from lipids fueling respiration up to NF 44-45. Quantification of total body composition revealed that lipid stores greatly influenced the calculations of MLB and therefore had profound underestimating effects on the mass-specific expression of numerous physiological measures through development.

Sequential changes in in vivo muscle and liver protein synthesis and plasma and tissue glutamine levels in sepsis in the rat

2001 ◽  
Vol 101 (3) ◽  
pp. 295-304 ◽  
Author(s):  
Michael J. O'LEARY ◽  
Colin N. FERGUSON ◽  
Michael J. RENNIE ◽  
Charles J. HINDS ◽  
John H. COAKLEY ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Total Protein ◽  
Muscle Protein ◽  
Muscle Protein Synthesis ◽  
Total Protein Content ◽  
Sham Operation ◽  
Liver Protein ◽  
Male Wistar Rats

We have investigated sequential changes in skeletal muscle and hepatic protein synthesis following sepsis, and their relationship to changes in circulating and tissue glutamine concentrations. Male Wistar rats underwent caecal ligation and puncture (CLP) or sham operation, with starvation, and were killed 24, 72 or 96 h later. A group of non-operated animals were killed at the time of surgery. Protein synthesis was determined using a flooding dose of l-[4-3H] phenylalanine, and glutamine concentrations were measured by an enzymic fluorimetric assay. Protein synthesis in gastrocnemius muscle fell in all groups. Gastrocnemius total protein content was reduced after CLP and at 72 and 96 h after sham operation. After CLP, protein synthesis was lower at 24 h, and total protein content was lower at 72 and 96 h, than in sham-operated animals. CLP was associated with increased liver protein synthesis at all time points, whereas there was no change after sham operation. Liver protein content did not change after CLP, but was lower at 72 and 96 h after sham operation than in non-operated animals. Plasma glutamine concentrations were reduced at 24 h after sham operation, and at 72 and 96 h after CLP. Muscle glutamine concentrations were reduced in all groups, with the decrease being greater following CLP than after sham operation. In the liver, glutamine concentrations were unchanged after CLP, but increased after sham operation. In rats with sepsis, decreases in muscle protein synthesis and content are associated with markedly reduced muscle glutamine concentrations. Plasma glutamine concentrations are initially maintained, but fall later. In liver, protein synthesis is increased, while glutamine concentrations are preserved. These results support a peripheral-to-splanchnic glutamine flux in sepsis.

Prediction of Human Acute Toxicity by the Hep G2/24-hour/Total Protein Assay, with Protein Measurement by the CBQCA Method

2005 ◽  
Vol 33 (3) ◽  
pp. 207-213 ◽  
Author(s):  
Paul J. Dierickx
Keyword(s):  
Protein Content ◽  
Total Protein ◽  
Total Protein Content ◽  
Human Toxicity ◽  
Hep G2 ◽  
Vitro Assay ◽  
Protein Assay ◽  
Lowry Method ◽  
Total Protein Assay

In our previously described Hep G2/24-hour/total protein assay, protein levels were measured by using the Lowry method. This assay was the best acute in vitro assay for the prediction of human toxicity within the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) study. In order to increase the MEIC data-base with a wider range of chemicals, we were interested in introducing the more practical 3-(4-carboxybenzoyl)-quinoline-2-carboxaldehyde (CBQCA) method for the quantification of the total protein content. Therefore, we investigated whether the same good results for the prediction of acute human toxicity would be obtained with the CBQCA method. The cells were treated for 24 hours, then cytotoxicity was determined by measuring the total protein content with CBQCA. The results were quantified by using the PI50c: the concentration (in mM) of test compound required to reduce the total protein content measured with the CBQCA-method by 50% as compared to the control cells. The results were compared with the PI50, the corresponding value when the Lowry method was used. A relatively low correlation was observed between PI50 and PI50c, reflecting the large and unexpected, differences when using the two protein assays. However, when comparing the log PI50c with the human toxicity, a correlation coefficient of r2 = 0.761 ( n = 44) was obtained for exactly the same series of MEIC chemicals. This value is clearly higher than that for the Lowry method ( r2 = 0.695). Compared to the Lowry method originally used, the Hep G2/24-hour/CBQCA total protein assay has the additional important advantage that it can be very easily adapted for large-scale analyses with robotic systems, including the on-line calculation of the results.

Total protein content and protein synthesis within pre-elongation stage bovine embryos

1998 ◽  
Vol 50 (2) ◽  
pp. 139-145 ◽  
Author(s):  
J.G. Thompson ◽  
A.N.M. Sherman ◽  
N.W. Allen ◽  
L.T. McGowan ◽  
H.R. Tervit
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Total Protein ◽  
Total Protein Content ◽  
Bovine Embryos

scholarly journals Surface topography of hydroxyapatite affects ROS17/2.8 cells response

2002 ◽  
Vol 16 (3) ◽  
pp. 209-215 ◽  
Author(s):  
Adalberto Luiz Rosa ◽  
Márcio Mateus Beloti ◽  
Richard van Noort ◽  
Paul Vincent Hatton ◽  
Anne Jane Devlin
Keyword(s):  
Cell Proliferation ◽  
Protein Content ◽  
Surface Topography ◽  
Cell Morphology ◽  
Total Protein ◽  
Cell Attachment ◽  
Maxillofacial Surgery ◽  
Total Protein Content ◽  
Alp Activity ◽  
Powder Pressing

Hydroxyapatite (HA) has been used in orthopedic, dental, and maxillofacial surgery as a bone substitute. The aim of this investigation was to study the effect of surface topography produced by the presence of microporosity on cell response, evaluating: cell attachment, cell morphology, cell proliferation, total protein content, and alkaline phosphatase (ALP) activity. HA discs with different percentages of microporosity (< 5%, 15%, and 30%) were confected by means of the combination of uniaxial powder pressing and different sintering conditions. ROS17/2.8 cells were cultured on HA discs. For the evaluation of attachment, cells were cultured for two hours. Cell morphology was evaluated after seven days. After seven and fourteen days, cell proliferation, total protein content, and ALP activity were measured. Data were compared by means of ANOVA and Duncan’s multiple range test, when appropriate. Cell attachment (p = 0.11) and total protein content (p = 0.31) were not affected by surface topography. Proliferation after 7 and 14 days (p = 0.0007 and p = 0.003, respectively), and ALP activity (p = 0.0007) were both significantly decreased by the most irregular surface (HA30). These results suggest that initial cell events were not affected by surface topography, while surfaces with more regular topography, as those present in HA with 15% or less of microporosity, favored intermediary and final events such as cell proliferation and ALP activity.

Development and regression of exercise-induced cardiac hypertrophy in rats

1979 ◽  
Vol 236 (2) ◽  
pp. H268-H272 ◽  
Author(s):  
R. C. Hickson ◽  
G. T. Hammons ◽  
J. O. Holoszy
Keyword(s):  
Cytochrome C ◽  
Protein Content ◽  
Total Protein ◽  
Female Rats ◽  
Heart Weight ◽  
Total Protein Content ◽  
Mitochondrial Marker ◽  
Time Intervals ◽  
Control Value ◽  
Exercise Induced

Adult female rats were exercised by daily swimming. All the increase in heart weight induced by the exercise occurred within 14 days and averaged 30%. The half times of the increases in heart weight and total protein content were about 4.5 days, whereas that of cytochrome c, which was used as a mitochondrial marker, was 6.5 days. The total amounts of DNA and of hydroxyproline in the heart, which were used to evaluate the degree of connective tissue hyperplasia, increased only slightly (8% and 10%, respectively). Other animals were subjected to the same swimming program for 21 days. Groups of rats were killed at various time intervals after stopping exercise. Heart weight, total protein content, and total cytochrome c content decreased rapidly initially, with 60% of the total regression of hypertrophy occurring during the first week. Thereafter, heart weight fell more gradually toward the sedentary control value. The hydroxyproline content of the heart, which was increased 10%, did not decrease during the regression of the hypertrophy.

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