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Biomedical subjects

R B Young

Publications and source records attributed to R B Young.

At least 37 records · Page 2Linked to original sources
PubMed1984

Leucine pools in normal and dystrophic chicken skeletal muscle cells in culture.

The specific radioactivity of [3H]Leu in the extracellular, intracellular, and Leu-tRNA pools of normal (white leghorn) and dystrophic (line 307) embryonic chick breast muscle cultures was analyzed as a function of equilibration time and extracellular Leu concentration (0.05-5 mM). The primary results were the following 1) [3H]Leu equilibrated to a constant specific radioactivity in the intracellular and Leu-tRNA pools within 2 min after addition to both normal and dystrophic cultures. 2) After equilibration, the extracellular [3H] Leu specific radioactivity in dystrophic cell culture medium was lower than that of medium exposed to normal cells (especially at low Leu concentrations), probably because of increased release of unlabeled Leu from the dystrophic cells as a result of faster protein breakdown. Accordingly, the specific radioactivities in the intracellular and the Leu-tRNA pools were also lower in dystrophic cells. 3) At 5 mM extracellular Leu, the specific radioactivity in the Leu-tRNA pool was approximately 40% lower than the specific radioactivity in the intracellular pool in both normal and dystrophic cells. Thus, high concentrations of extracellular Leu cannot be used to "flood out" reutilization of unlabeled Leu (released by protein degradation) during protein synthesis. 4) At 5.0 mM extracellular Leu, the specific radioactivity of [3H]Leu in the intracellular pool was comparable to that in the extracellular pool in normal and dystrophic cells; however, the specific radioactivity of Leu-tRNA (i.e. the immediate precursor to protein synthesis) was only 55-65% of the extracellular specific radioactivity in normal and dystrophic cells. In conclusion, reutilization of Leu from protein degradation is higher in dystrophic muscle cell cultures than in normal muscle cell cultures, and accurate rates of protein synthesis in cell cultures can only be obtained if specific radioactivity of amino acid in tRNA is measured.

Animals↗
PubMed1984

Elevated ammonia release from dystrophic chicken muscle cell and fibroblast cultures.

The activity of adenosine monophosphate (AMP)-aminohydrolase, the major NH3-producing enzyme in skeletal muscle, was approximately 35% lower in 7-day dystrophic muscle cell cultures than in normal muscle cell cultures. However, the release rate of NH3 from dystrophic muscle cells was 45% higher than that from normal muscle cells. The reasons for this apparent discrepancy are not clear. To determine indirectly if deamination of amino acids from protein degradation contributed to NH3 release, cells were incubated with 100 micrograms/ml of the protease inhibitor, leupeptin. Leupeptin reduced the rate of NH3 release by only 18.8% in normal muscle cells and 16% in dystrophic muscle cells. The release of NH3 was also higher from dystrophic chicken fibroblast cultures.

Adenosine Deaminase↗
PubMed1984

Myosin heavy chain concentration, synthesis rate and degradation rate in normal and dystrophic chicken muscle cells in culture.

Myosin heavy chain concentrations, synthesis rates and degradation rates were studied in muscle cell cultures prepared from the breast muscle of 13-day normal (white leghorn) and dystrophic (line 307) chicken embryos. Muscle cells were studied after 7 days in culture, at which time they had reached a steady state with respect to myofibrillar protein synthesis and degradation. The quantity of myosin heavy chain was 10.5 +/- 0.9 micrograms/culture (n = 32) in normal cells and 8.10 +/- 1.2 micrograms/culture (n = 31) in dystrophic cells. However, the myosin heavy chain synthesis rate was 23 500 molecules/min/nucleus in normal cells and 39 900 molecules/min/nucleus in dystrophic cells, as determined by pulse labeling with [3H]leucine and measurement of the specific radioactivity of tRNA precursor pools. Myosin heavy chain half-lives rates were calculated to be 30.6 h in normal cells and 15.6 h in dystrophic cells when corrections were made for reutilization of [3H]leucine. Thus, dystrophic muscle cultures accumulate less myosin heavy chain, despite their faster synthesis rate, because of faster degradation of myosin heavy chain.

Animals↗
PubMed1983

Control of respiratory pattern in conscious dog: effects of heat and CO2.

We measured tidal volume (VT) and inspiratory (TI) and expiratory (TE) durations in five conscious tracheostomized dogs breathing air or 5% CO2 in air either at normal (20 degrees C) or elevated (30 degrees C) ambient temperatures. Respiratory frequency ranged between 16 and 333/min due to changes in both TI and TE. During panting TI exceeded TE. During air inhalation instantaneous ventilation (V) spontaneously ranged from 100 to 1,600 ml . kg-1 . min-1. Hypercapnia, heat stress, or both, increased this range of V by increasing maximum V, primarily due to increases in mean inspiratory flow. Under these conditions, changes in TI accounted for more of the spontaneous changes in breath duration. During inhalation of air and 5% CO2, a positive correlation between VT and TI was obtained for TI between 0.13 and 1.05 s; above 1.05 s VT decreased. Heat stress increased VT at a given TI. We suggest that either the decay rate or position of the inspiratory off-switch threshold curve (Clark and von Euler, J. Physiol. London 222: 267, 1972) varies in conscious dogs. Shifts in either the reset (onset) value or decay rate of the curve yield a positive correlation between VT and TI. This modification to the Clark-von Euler model implies that the primary effect of anesthesia on respiratory control is fixation of the inspiratory off-switch threshold curve.

Animals↗
PubMed1982

Myocardial pH during regional ischemia: evaluation of a fiber-optic photometric probe.

The relationship between the decrease in intramyocardial extracellular pH and the degree of stenosis of the left anterior descending (LAD) coronary artery was studied in eight dogs pretreated with propranolol. Intramyocardial pH was measured with a miniature glass pH electrode and with a new photometric pH probe that uses fiber-optic filaments to measure the color change of an indicator substance in a small permeable chamber. The LAD was cannulated and perfused from the axillary artery. Cannula flow was measured with an electromagnetic flow probe, and regional myocardial blood flow (RMBF) was measured with radioactive microspheres before and at the end of a period of critical stenosis, 2/3 reduction of flow, or total occlusion of the LAD cannula. In the region of the glass electrode, the mean RMBF (+/-SE) decreased by 16.3 +/- 3.3, 52.7 +/- 7.3, and 84.8 +/- 6.5% during the three levels of stenosis, and the pH correspondingly decreased by 0.05 +/- 0.01, 0.29 +/- 0.10, and 0.94 +/- 0.17 units. In the region of the photometric probe, the RMBF decreased 19.1 +/- 1.3, 47.2 +/- 6.7, and 84.3 +/- 6.0%, and the pH decreased by 0.05 +/- 0.02, 0.14 +/- 0.04, and 0.76 +/- 0.18 units. There was no statistically significant difference between the two types of pH sensor.

Animals↗
PubMed1982

Cell differentiation, protein synthesis rate and protein accumulation in muscle cell cultures isolated from embryos of layer and broiler chickens.

Muscle cell cultures were prepared from the leg muscle of 12-d layer and broiler chicken embryos. Cultures were then compared over a 10-d period from their capacity to differentiate into multinucleated myotubes and to synthesize and accumulate protein. Differentiation was qualitatively similar in the two cell types as evidenced by myoblast fusion that occurred rapidly during the first 2 d and remained essentially constant between d 3 and 10. However, several quantitative differences were observed. Even though the number of myotubes per culture was comparable between layers and broilers throughout development, layer muscle cultures usually exhibited a higher percentage fusion and more myonuclei per culture than broiler muscle cultures. Additionally, the nuclear density (i.e., the number of nuclei per myotube segment) was approximately 25% greater in layer cultures than in broilers between d 2 and 10 in culture. The rate of incorporation of 3H-leucine into total protein during pulse labeling experiments was comparable in muscle cultures of layers and broilers; however, broiler muscle cells accumulated approximately 40% more total protein per nucleus between d 6 and 10. Myosin heavy chain synthesis rate was higher in layer than in broiler muscle cultures, but broiler muscle cultures accumulated approximately 30% more myosin heavy chain than layers between d 6 and 10. The half-life of myosin heavy chain was 45 h in layer muscle cultures and 103 h in broiler muscle cultures. Thus, the capacity of broiler cells to accumulate more muscle protein was primarily due to a drastically slower protein breakdown rate.

Animals↗
PubMed1981

Metabolism of myosin heavy chain in steady-state chick skeletal muscle cultures.

Synthesis, accumulation and breakdown of the 200000-mol.wt. heavy subunit of myosin were analysed over an 11 day period in muscle cell cultures isolated from the leg muscle of 12-day chick embryos. Muscle cells accumulated myosin heavy chain rapidly from days 2 to 5 and maintained a maximum, constant myosin-heavy-chain concentration between days 7 and 11. Myosin-heavy-chain content and breakdown rate were compared in steady-state muscle cultures grown either in the presence of an optimum batch of horse serum (control) or in the presence of horse serum that had been pre-selected for its ability to inhibit several-fold the rate of synthesis of myosin heavy chain (inhibitory). The quantity of myosin heavy chain in the inhibited cultures was decreased in direct proportion to the decrease in the rate of synthesis of myosin heavy chain; however, the half-lives of myosin heavy chain (control, 17.7h; inhibitory, 17.0h) were virtually identical. In contrast, the absolute rate of breakdown of myosin heavy chain, expressed as molecules/min per nucleus, was approx. 5-fold lower in the inhibited cultures (4.3 X 10(3) molecules/min per nucleus) than in the control cultures (21.7 X 10(3) molecules/min per nucleus). Thus, inhibition of myosin-heavy-chain synthesis in this case was accompanied by diminished myosin-heavy-chain concentration and absolute breakdown rate at the altered steady state, but relative myosin-heavy-chain breakdown rates were unchanged.

Animals↗
PubMed1981

Stimulation of myosin heavy chain synthesis in steady-state muscle cultures by the ionophore, A23187, requires transcription of messenger RNA.

After approximately one week in culture, embryonic chick skeletal muscle cells are at a steady state with respect to myosin heavy chain (MHC) concentration and synthesis rate. Muscle cells normally synthesize MHC at a maximum rate of 2.3 X 10 4 MHC/min/nucleus and contain approximately 3 X 10 7 MHC/nucleus. These cells also contain approximately 3500 copies/nucleus of MHC mRNA associated with polysomes and 1600 copies/nucleus of MHC mRNA localized in the nonpolysomal fraction. To determine if nonpolysomal MHC mRNA in mature muscle cultures could be recruited into active translation complexes when MHC synthesis was stimulated, muscle cultures were treated with the Ca 2+ ionophore, A23187 (0-1 micro M). The MHC synthesis rate was stimulated by 25 to 50% relative to stimulation of the rate of total protein synthesis in the presence of A23187, but this stimulation was blocked when 10 microgram/ml actinomycin D was also present. These results suggest that even though 30% of MHC mRNA is not actively engaged in MHC synthesis in mature muscle cultures, stimulation of MHC synthesis by A23187 results from transcription of new MHC mRNA rather than from utilization of pre-existing mRNA.

Animals↗
PubMed1979

Myosin synthesis in embryonic chicken fibroblasts.

The rate of constitutive myosin synthesis was measured in cultures of replicating embryonic chicken skin fibroblasts by pulse labeling with [3H]leucine. These cells synthesized the 200,000-dalton heavy chain of myosin (MHC) at a rate of 3.2 x 10(3) molecules/cell/min. Additionally, an independent estimate of the MHC synthesis rate needed to maintain a constant level of constitutive MHC/cell was calculated from total protein content, percentage MHC, fibroblast doubling time, and MHC half-life. This calculated rate of approximately 2.9 x 10(3) molecules/cell/min was in close agreement with the measured rate. By comparison, the synthesis rate of myofibrillar MHC in fully activated muscle cell cultures was approximately 2.9 x 10(4) molecules/nucleus/min.

Animals↗