Investigation of actin in Tetrahymena cells. A comparison with skeletal muscle actin by a devised two-dimensional gel electrophoresis method

Total protein constituents of Tetrahymena thermophila strain B1868 III were studied by two-dimensional agarose-polyacrylamide gel electrophoresis to detect actin among the constituents. In the attempts to prepare a whole-cell extract of Tetrahymena, it was found that protease activity in the extract was so high that high molecular components were quickly digested with the endogenous protease into small peptides unless the homogenization and heat-treatment in a sodium dodecylsulfate solution were performed within 5 s. It was eventually found that employment of 8 M guanidine hydrochloride (HCl) in the homogenization of cells perfectly prevented the degradation of protein components, even through a long preparation procedure. A devised two-dimensional agarose-polyacrylamide gel electrophoresis of the guanidine HCl extract gave a 'protein map' on which most proteins were located in their respective positions, including proteins with more than 200,000 mol. wt. Addition of rabbit skeletal muscle actin on the protein map revealed that no protein with isoelectric point and molecular weight identical with those of the actin was contained in the whole Tetrahymena extract, suggesting that Tetrahymena actin may have characteristics far different from those of skeletal muscle actin.     

Relationships among the muscle strength properties as assessed through various tests and variables

We tested the hypotheses that the individual strength properties depend on the applied test and the variable extracted, rather than on the muscle group tested. Flexor and extensor muscles acting in the knee and elbow joint were tested in 58 participants. The standard strength test (SST; based on sustained maximum contraction) and alternating consecutive maximum contractions (ACMCs; alternating contractions of antagonistic muscles) performed under static conditions were separately applied to provide the maximum force (F) and the rate of force development (RFD) of each tested muscle. The principal component analysis applied on all 16 variables revealed three factors that explained 85.5% of the total variance. Contrary to our hypotheses, the individual factors were loaded with the variables recorded from individual muscles, rather than with either the particular variables or tests. The present findings suggest that recording both F and RFD in routine strength testing procedures could be redundant since they may assess the same strength property of the tested muscle. In addition, ACMC may be a feasible alternative to SST since it could assess the same strength properties from two antagonist muscles through a single trial, while being based on relatively low and transient forces.     

The difference in output properties between dominant and nondominant limbs as measured by various muscle function tests

Recently, right and left output properties exerted from specific muscle groups have been evaluated using special measurement devices such as an isokinetic dynamometer. However, it remains unclear whether the coach can properly evaluate muscle function corresponding to lateral specificity in athletes. This study aimed to examine the different output properties between the dominant (D) and nondominant (ND) upper limbs as measured by muscle function tests with various muscle contraction patterns. Fifteen right-handed young men participated in this study. Each subject carried out isometric, isokinetic, and isotonic muscle power tests by elbow flexion with right and left arms. When calculating the laterality index, the laterality of the isotonic test (1.17) was the highest. In all tests, significant correlations were found between the measurements of the D and ND limbs. The isometric test was the highest (r = 0.93), followed by the isokinetic test (r = 0.66-0.83) and the isotonic test (r = 0.55). To examine the ratio of the laterality of measurements provided by each muscle function test, each measurement was converted to a standard score (Z-score). There were significant differences between D and ND limbs in the isometric (D:ND = 55.0:45.0) and the isotonic (54.1:45.9) tests but not in the isokinetic test (60°·s?1, 51.4:48.6; 180°·s?1, 50.7:49.2; 300°·s?1, 51.8:48.2). Particularly, the D (right) limb exerted greater muscle power in the isometric and the isotonic tests than in the isokinetic test. Occupational therapists or strength and conditioning professionals should understand that the D-ND differences shown by these muscle function tests may differ because of measurement conditions.     

DNA repair assays as tests for environmental mutagens: A report of the U.S. EPA gene-tox program

A literature review was undertaken to determine the usefulness of DNA repair assays, other than unscheduled DNA synthesis, as screening techniques for mutagenic carcinogens. 92 reports were found to contain useful data for 49 chemicals using 6 techniques, namely, (1) cesium chloride equilibrium density gradients to study repair replication, (2) benzoylated naphthoylated diethylaminoethyl cellulose columns to study repair replication, (3) 313-nm irradiation of DNA containing bromodeoxyuridine to study repair replication, (4) alkaline elution to study repair of single-strand breaks and crosslinks, (5) alkaline sucrose gradients to study repair of single-strand breaks, and (6) direct assays for removal of adducts from DNA. Almost all of the 49 chemicals studied were known mutagens or carcinogens and/or known inducers of DNA repair. 9 compounds failed to elicit DNA repair by at least 1 assay technique, and at least 3 of these were not tested by the most appropriate and sensitive method. Nevertheless, although valid for studying repair phenomena in eukaryotic cells, these assays are not considered useful for screening. They are time-consuming, expensive, and/or require highly specialized skills. Despite the high frequency of positive reports, it is obvious from the literature that repair assays will fail to detect, or will detect with low efficiency, those agents whose main action is either intercalation or induction of strand breaks. For these and other reasons, DNA repair as a basis for screening for mutagenic carcinogens is not considered to be a useful concept.     

Amphotericin B-induced ion flux is markedly attenuated in phosphatidylglycerol membrane as evidenced by a newly devised fluorometric method

The effect of phospholipid head group on the membrane-permeabilizing activity of amphotericin B (AmB) was examined using 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) liposomes and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG) liposomes. The activity of AmB was evaluated as K⁺ influx measured as pH change inside liposomes by fluorescent measurements of 2′,7′-bis(carboxyethyl)-4 or 5-carboxyfluorescein (BCECF). AmB showed prominent permeability in POPC liposomes, whereas hardly inducing ion flux in POPG membrane. POPC added to POPG liposomes as a minor constituent markedly enhanced membrane permeability, indicating the importance of a phosphonocholine group of PC for the drug’s activity.     

Extensibility of the myofilaments in vertebrate skeletal muscle as revealed by stretching rigor muscle fibers

The extensibility of the myofilaments in vertebrate skeletal muscle was studied by stretching glycerinated rabbit psoas muscle fibers in rigor state and examining the resulting extension of sarcomere structures under an electron microscope. Although stretches applied to rigor fibers produced a successive yielding of the weakest sarcomeres, the length of the remaining intact sarcomeres in many myofibrils was fairly uniform, being definitely longer than the sarcomeres in the control, nonstretched part of rigor fibers. The stretch-induced increase in sarcomere length was found to be taken up by the extension of the H zone and the I band, whereas the amount of overlap between the thick and thin filaments did not change appreciably with stretches of 10-20%. The thick filament extension in the H zone was localized in the bare regions, whereas the thin filament extension in the I band appeared to take place uniformly along the filament length. No marked increase in the Z-line width was observed even with stretches of 20-30%. These results clearly demonstrate the extensibility of the thick and thin filaments. The possible contribution of the myofilament compliance to the series elastic component (SEC) in vertebrate skeletal muscle fibers is discussed on the basis of the electron microscopic data and the force-extension curve of the SEC in rigor fibers.     

Isotonic and isoelectric endurance tests for the upper trapezius muscle

As an alternative to a conventional endurance electromyogram (EMG) test for assessment of muscle capacity in the upper trapezius muscle (isotonic test, IT), an isoelectric test (IE) has been investigated. Nine subjects performed the two endurance tests. The tests were performed with a straight horizontal arm in the plane of the scapula. In IE, EMG amplitude was fed back to the subject and the subject was instructed to maintain a constant EMG activity equal to the level with the arm unsupported. Subjective ratings of local fatigue were obtained during the experiments. The EMG recordings from both tests were analysed for the root mean square value as well as according to the mean power frequency (MPF) technique. All the subjects endured maximal 15 min IE while in IT the median endurance was 11.9 min. Average subjective ratings of perceived fatigue increased more in IT compared to IE. The average normalised MPF curve from IT increased while that from IE was unaffected. There was a significant difference between the MPF results at 6 and 8 min. It is noteworthy that the MPF was higher in IT in spite of a higher accumulated biomechanical load. It was concluded from these studies that the interpretation of decreased MPF as an indicator of increased local muscle fatigue is doubtful at low contraction levels. It is suggested that an IE is a more appropriate method for the functional evaluation of low threshold motor units of the upper trapezius muscle in research into occupational disorders. Accepted: 15 October 1996     

Nodular sarcoid myositis of skeletal muscle diagnosed by fine needle aspiration biopsy. A case report

BACKGROUND: Symptomatic striated muscle involvement in sarcoidosis is rare. Muscle biopsy is usually required for the diagnosis. Fine needle aspiration biopsy (FNAB) has been successfully used in diagnosing soft tissue lesions. To the best of our knowledge, FNAB of sarcoid myositis has not been reported. CASE: A 31-year-old, black female with a history of sarcoidosis presented with an enlarging, painful, left calf mass. Infected thrombi were suspected. FNAB showed numerous loosely arranged epithelioid histiocytes, multinucleated giant cells and skeletal muscle cells. The overall cytologic picture was that of granulomatous myositis. The cytologic features coupled with the patient's history and magnetic resonance imaging findings suggested sarcoid myositis. Subsequent muscle biopsy showing noncaseating granulomata and negative stains for organisms confirmed the diagnosis of nodular sarcoid myositis. CONCLUSION: Nodular sarcoid myositis can be suggested by FNAB cytology in a patient with a past history of sarcoidosis.