Health practices of educators participating in an Idaho wellness conference: one year follow-up.

The Great Potato Health Conference (GPHC), which convenes annually in Idaho, is one of more than 25 statewide wellness conferences for K-12 school personnel. This follow-up study assessed the health practices of conference participants one year following the GPHC. A 25-item health behavior instrument was administered to 68 school district personnel prior to attending the GPHC and again three months, six months, and one year following the conference. Data were analyzed using a repeated measures one-way factorial analysis of variance to test for differences in the health practices of participants over time. Results indicated statistically significant main effects for both previous conference attendance and time of measurement. No statistically significant interaction effect occurred between previous conference attendance and time of measurement.

Desmin is present in proliferating rat muscle satellite cells but not in bovine muscle satellite cells.

The presence of desmin was characterized in cultured rat and bovine satellite cells and its potential usefulness as a marker for identifying satellite cells in vitro was evaluated. In primary cultures, positive immunohistochemical staining for desmin and skeletal muscle myosin was observed in rat and bovine myotubes. A small number of mononucleated cells (20% of rat satellite cells and 5% of bovine satellite cells) were myosin-positive, indicative of post-mitotic differentiated myocytes. In bovine satellite cell cultures 13% of the mononucleated cells were desmin-positive, while 84% of the mononucleated cells in rat satellite cell cultures were desmin-positive. Rat satellite cell mass cultures and bovine satellite cell clonal density cultures were pulsed with 3H-thymidine, and autoradiographic data revealed that greater than 94% of dividing rat cells were desmin-positive, suggesting that desmin is synthesized in proliferating rat satellite cells. However, no desmin was seen in cells that incorporated labeled thymidine in bovine satellite cell clones. Analysis of clonal density cultures revealed that only 14% of the mononucleated cells in bovine satellite cell colonies were desmin-positive, whereas 98% of the cells in rat satellite cell colonies were desmin-positive. Fibroblast colonies from both species were desmin-negative. In order to further examine the relationship between satellite cell differentiation and desmin expression, 5-bromo-2'-deoxyuridine (BrdU) was added to culture medium at the time of plating to inhibit differentiation. Fusion was inhibited in rat and bovine cultures, and cells continued to divide. Very few desmin-positive cells were found in bovine cultures, but greater than 90% of the cells in rat cultures stained positive for desmin. The presence of desmin and sarcomeric myosin was also evaluated in regenerating rat tibialis anterior five days after bupivacaine injection. In regenerating areas of the muscle many desmin-positive cells were present, and only a few cells stained positive for skeletal muscle myosin. Application of desmin staining to rat satellite cell growth assays indicated that rat satellite cells cultured in serum-containing medium were contaminated with fibroblasts at levels that ranged from approximately 5% in 24 hr cultures to 15% in mature cultures. In defined medium 4 day cultures contain approximately 95% to 98% desmin-positive satellite cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Calyculin-A increases the level of protein phosphorylation and changes the shape of 3T3 fibroblasts.

Calyculin-A, an inhibitor of type 1 and 2A phosphatases, was applied extracellularly to 3T3 fibroblasts. At 0.1 microM, calyculin-A caused a marked increase in protein phosphorylation in both the cytosolic and insoluble cellular fractions. This effect was independent of external Ca2+. An immunoprecipitate, formed with an antibody to myosin, contained several cytoskeletal components. Increased phosphorylation following treatment with calyculin-A was observed in vimentin, the 20-kD myosin light chain, and an unidentified 440-kD component. An enhanced level of vimentin phosphorylation was found in intermediate filament preparations from treated cells. Calyculin-A also caused marked shape changes of 3T3 cells. Within minutes after addition of calyculin-A (0.1 microM) cells became rounded and lost attachment to the substratum. Stress fibers, intermediate filaments, and microtubules, prominent in the attached control cells, were not evident in the rounded cells. Shape changes were reversible and after removal of calyculin-A the rounded cells attached to the substratum, resumed a flattened shape, and were active mitotically. In the cells treated with calyculin-A an unusual "ball-like" structure was observed with transmission electron microscopy. This unique structure was 2-3 microM in diameter and was located close to the nucleus. The use of calyculin-A adds further support to the idea that cell shape is controlled, at least in part, by concerted actions of a kinase-phosphatase couple.

Regulation of satellite cells during skeletal muscle growth and development.

Satellite cells are myogenic cells attributed with the role of postnatal growth and regeneration in skeletal muscle. Following proliferation and subsequent differentiation, these cells will fuse with one another or with the adjacent muscle fiber, thereby increasing myonuclei numbers for fiber growth and repair. The potential factors which could regulate this process are many, including exercise, trauma, passive stretch, innervation, and soluble growth factors. Three classes of growth factors in particular (fibroblast growth factor, insulin-like growth factor, and transforming growth factor-beta) have been studied extensively with respect to their effects on satellite cell proliferation and differentiation in culture. Fibroblast growth factor has been shown to stimulate proliferation but depress differentiation. Insulin-like growth factor stimulates both proliferation and differentiation, although the latter to a much greater degree. Transforming growth factor-beta slightly depresses proliferation but inhibits differentiation. When administered in combination, these factors can induce satellite cell activities in culture which mimic those typical of satellite cells found in vivo in growing, regenerating, or healthy mature muscle. Alterations in the concentrations of these growth factors in the muscle environment as well as alterations in the cell's sensitivity or responsiveness to these factors represent potential mechanisms for regulating satellite cell activity in situ.

Fatigability of rat hindlimb muscle: associations between electromyogram and force during a fatigue test.

1. An experimental protocol designed to assess fatigability in motor units (Burke, Levine, Tsairis & Zajac, 1973) has been applied to the whole muscles of anaesthetized adult rats, and the association between the electromyogram (EMG) and force was monitored over the course of the test. 2. Both test muscles (soleus and extensor digitorum longus) exhibited a wide range of fatigability, which was defined as the decline in isometric peak force at 6 min, such that the data could be separated into five levels of fatigability. Fatigue indices for each test muscle were distributed across three levels. 3. The EMG was quantified with four measures of amplitude, four of duration, and one interaction term (area). Correlation analyses indicated that the EMG was adequately represented by one measure of amplitude (absolute amplitude), one of duration (peak-to-peak duration) and area. The best single measure was area. 4. The EMG-force associations for soleus varied markedly among its three fatigability groups. In contrast, over the course of the test, all three extensor digitorum longus groups displayed qualitatively similar EMG-force associations. 5. Multiple regression analyses indicated that the EMG parameters were able to predict peak force better for extensor digitorum longus than for soleus. Furthermore, for both test muscle, the prediction was best for the most fatigable group. 6. The associations between EMG and force exhibited three patterns for the two test muscles and three levels of fatigability. These differences suggested variation in the mechanisms, related to both fibre-type composition and susceptibility to fatigue, that dictate the performance elicited by this particular stimulus regimen. The mechanisms seem to include both intracellular and transmission processes.

Coexistence of twitch potentiation and tetanic force decline in rat hindlimb muscle.

An experimental protocol designed to assess fatigability in motor units has been applied to two hindlimb muscles of anesthetized adult rats to study the effects of whole-muscle fatigue on the isometric twitch. Both soleus and extensor digitorum longus exhibited a linear relationship between fatigability (i.e., force decline after a 360-s fatigue test) and the magnitude of the twitch force following the fatigue test. Twitch force after the fatigue test was potentiated (i.e., greater than the value before the fatigue test) in many muscles, despite the development of considerable fatigue. This coexistence of fatigue and twitch potentiation was observed in 7% (5/70) of soleus and 48% (31/64) of extensor digitorum longus muscles. The coexistence was exhibited only by the least fatigable muscles of the fast-contracting extensor digitorum longus. The extensor digitorum longus muscles that did not exhibit twitch potentiation probably experienced a higher proportion of muscle-fiber inactivation, such as due to failure of neuromuscular propagation, that was induced by the fatigue regimen.

Fatigue-related changes in neuromuscular excitability of rat hindlimb muscles.

Brief repetitive trains of supramaximal nerve stimulation produce intermittent muscle activation and, in time, a progressive decline in force (i.e., neuromuscular fatigue) and depression of the electromyogram (EMG). These changes may include within-train reductions in EMG due to a failure of neuromuscular propagation. The aim of the present study was to investigate changes in EMG during a 360-second stimulus regimen designed to fatigue soleus and extensor digitorum longus muscles of anesthetized rats by activating the muscle with repetitive trains of 40 Hz stimuli. Measurements included peak force for each tetanus, variation of the within-train EMG (coefficient of variation for area), and magnitude of the first EMG waveform (area) of each train. Fatigue was characterized as the relative decline in force over the course of the test. The responses of the test muscles were categorized, based on an absolute scale of fatigability, into five groups: potentiated, nonfatigable, low fatigability, intermediate fatigability, and high fatigability. Fatigable muscles (low, intermediate, and high fatigability groups) demonstrated a decreased EMG magnitude and an increased EMG-area variation with repetitive activation. This increased variation, however, was nonmonotonically related to fatigability such that the least and most fatigable muscles had the smallest within-train EMG variation. We suggest that these data can be explained by considering the EMG (compound muscle action potential) as a stochastic process that represents a composite of single-fiber events (axonal to sarcolemmal transmission) with variable probabilities.

Diffusion characteristics of beta-aminopropionitrile in peripheral nerve.

beta-Aminopropionitrile (beta-APN), a lathyrogen, alters the physical characteristics of fibrous scar tissue and as such may have potential clinical use in treatment of injured spinal cord and peripheral nerve by reducing the physical barrier to axon regeneration. For beta-APN to exert its lathyrogenic effect, it must permeate the injury site and gain access to the developing collagenous scar. To investigate the diffusion characteristics, beta-[14C]APN solution was applied as an immersion bath to rat sciatic nerve using both in vivo and in vitro preparations for intervals of 15 to 90 min. The four experimental groups studied were (a) intact nerve, (b) hemisected nerve, (c) nerve with epineurium removed, and (d) nerve with both epineurium and perineurium removed. The isotope labeling index determined by autoradiography and scintillation counting indicated the perineurium as the primary barrier to significant diffusion of beta-APN in normal nerve. When perineurium was incised or removed, beta-APN entered the endoneurial matrix. beta-APN concentration in the epineurium and perineurium increased with increasing bathing time in vitro; but it decreased markedly after 15 min of in vivo bathing. These findings indicate that topical application of beta-APN to injured peripheral nerve would be a successful method of exposing fibrogenic intraneural tissue to the inhibitory effect on lysyl oxidase enzymes. Continuous application, however, will be necessary because of the rapid beta-APN removal documented in the vivo preparation.