The influence of drag on human locomotion in water.

Propulsion in water requires a propulsive force to overcome drag. Male subjects were measured for cycle frequency, energy cost and drag (D) as a function of velocity (V), up to maximal V, for fin and front crawl swimming, kayaking and rowing. The locomotion with the largest propulsive arms and longest hulls traveled the greatest distance per cycle (d/c) and reached higher maximal V. D while locomotoring increased as a function of V, with lower levels for kayaking and rowing at lower Vs. For Vs below 1 m/s, pressure D dominated, while friction D dominated up to 3 m/s, after which wave D dominated total D. Sport training reduced the D, increased d/c, and thus lowered C and increased maximal V. Maximal powers and responses to training were similar in all types of locomotion. To minimize C or maximize V, D has to be minimized by tailoring D type (friction, pressure or wave) to the form of locomotion and velocity.

An energy balance of front crawl.

With the aim of computing a complete energy balance of front crawl, the energy cost per unit distance (C = Ev(-1), where E is the metabolic power and v is the speed) and the overall efficiency (eta(o) = W(tot)/C, where W(tot) is the mechanical work per unit distance) were calculated for subjects swimming with and without fins. In aquatic locomotion W(tot) is given by the sum of: (1) W(int), the internal work, which was calculated from video analysis, (2) W(d), the work to overcome hydrodynamic resistance, which was calculated from measures of active drag, and (3) W(k), calculated from measures of Froude efficiency (eta(F)). In turn, eta(F) = W(d)/(W(d) + W(k)) and was calculated by modelling the arm movement as that of a paddle wheel. When swimming at speeds from 1.0 to 1.4 m s(-1), eta(F) is about 0.5, power to overcome water resistance (active body drag x v) and power to give water kinetic energy increase from 50 to 100 W, and internal mechanical power from 10 to 30 W. In the same range of speeds E increases from 600 to 1,200 W and C from 600 to 800 J m(-1). The use of fins decreases total mechanical power and C by the same amount (10-15%) so that eta(o) (overall efficiency) is the same when swimming with or without fins [0.20 (0.03)]. The values of eta(o) are higher than previously reported for the front crawl, essentially because of the larger values of W(tot) calculated in this study. This is so because the contribution of W(int) to W(tot )was taken into account, and because eta(F) was computed by also taking into account the contribution of the legs to forward propulsion.

Energy balance of human locomotion in water.

In this paper a complete energy balance for water locomotion is attempted with the aim of comparing different modes of transport in the aquatic environment (swimming underwater with SCUBA diving equipment, swimming at the surface: leg kicking and front crawl, kayaking and rowing). On the basis of the values of metabolic power (E), of the power needed to overcome water resistance (Wd) and of propelling efficiency (etaP=Wd/Wtot, where Wtot is the total mechanical power) as reported in the literature for each of these forms of locomotion, the energy cost per unit distance (C=E/v, where v is the velocity), the drag (performance) efficiency (etad=Wd/E) and the overall efficiency (etao=Wtot/E=etad/etaP) were calculated. As previously found for human locomotion on land, for a given metabolic power (e.g. 0.5 kW=1.43 l.min(-1) VO2) the decrease in C (from 0.88 kJ.m(-1) in SCUBA diving to 0.22 kJ.m(-1) in rowing) is associated with an increase in the speed of locomotion (from 0.6 m.s(-1) in SCUBA diving to 2.4 m.s(-1) in rowing). At variance with locomotion on land, however, the decrease in C is associated with an increase, rather than a decrease, of the total mechanical work per unit distance (Wtot, kJ.m(-1)). This is made possible by the increase of the overall efficiency of locomotion (etao=Wtot/E=Wtot/C) from the slow speeds (and loads) of swimming to the high speeds (and loads) attainable with hulls and boats (from 0.10 in SCUBA diving to 0.29 in rowing).

Synthesis of leukotrienes by freshly harvested endothelial cells.

Endothelial cells produce endothelin, a powerful vasoconstrictor. We report the release of additional vasoconstrictor material in conditional filtrate from freshly harvested cells, which we identified as leukotrienes by radioimmunoassay (RIA) and by high pressure liquid chromatography (HPLC). The material was collected in cell free filtrates by superfusion of freshly harvested bovine endothelial cells attached to cytodex-3 microcarrier beads. Cells and beads form a dense network on filter paper permitting collection of cell free filtrate. The amount of leukotrienes in conditioned filtrate was 158 +/- 21 picograms/million cells. The calcium ionophore A23187 stimulated the release of leukotrienes (392.0 +/- 47.6). The peak of leukotriene production occurred within an hour after incubation of cells slowly declining thereafter. Conditioned filtrate to which indomethacin had been added caused coronary vasoconstriction in the perfused rat heart preparation, as did synthetic leukotrienes C4, D4 and E4. It was found by RIA and HPLC that some of the constrictor effect of conditioned filtrate derived from leukotrienes.

Membrane function and vascular reactivity.

This communication examines the possibility that nitric oxide (NO) production by endothelial cells results from changes in cell membrane fluidity. Lysophosphatidylcholine (LPC) alters fluidity of the endothelial cell membranes causing vascular relaxation. Through membrane alterations LPC influences function of a number of membrane receptors and modulates enzyme activity. As a result of detergent action, lysophosphatidylcholine (LPC) causes activation of guanylate cyclase, stimulates sialyltransferase and regulates protein kinase C activity. It has already been demonstrated that ionic detergents, such as Triton X-100 also cause vascular relaxation, possibly induced by NO production from endothelial cells. It is postulated that production of nitric oxide results from changes in membrane viscosity; this may represent a mechanism for its regulation in biological systems.

The production of coronary vasoconstrictor substances by freshly harvested endothelial cells.

The effects of cell free superfusates from freshly harvested bovine endothelial cells attached to microcarrier beads on the isolated rabbit and rat heart and on superfused rabbit jugular veins were observed. Cell free conditioned filtrates from freshly harvested cells caused marked diminution in coronary flow and cardiac output in the isolated rabbit heart; in the perfused rat heart an increase in coronary perfusion pressure and a decline in left ventricular systolic tension and maximal left ventricular contractility (dP/dt) were recorded. Marked differences were found between changes induced by conditioned filtrate as compared to synthetic endothelin. Endothelin as present in conditioned filtrate could not account for the pronounced effect on coronary perfusion pressure, dp/dt and cardiac output induced by conditioned filtrate; more than one hundred times that of synthetic endothelin was needed to achieve comparable cardiodynamic effects. This suggested that additional non-prostanoid vasoconstrictor substance or substances are produced by freshly harvested endothelial cells. This conclusion was supported by the observation that BQ-123, a specific inhibitor of endothelin A (ETA) receptor significantly prevented contractions by endothelin, while failing to inhibit those induced by freshly harvested endothelial cells. These constrictor substances may be leukotrienes.

Changes in myosin heavy-chain isoform synthesis of chronically stimulated rat fast-twitch muscle.

Chronic low-frequency stimulation was used for studying the adaptive potential of rat fast-twitch muscle to increased neuromuscular activity. The sequential exchange of myosin heavy chain isoforms HCIIb with HCIId and HCIIa was studied at the translational level using an in-vivo-labeling technique with [35S]methionine. Alterations in heavy chain isoform synthesis, i.e. a decrease in the labeling of HCIIb concomitant with an enhanced labeling of HCIId/IIa, were detectable already two days after the onset of stimulation. This time course corresponds to the previously observed alterations in the amounts of HCIIb and HCIIa mRNAs. However, significant changes in the relative protein amounts of HCIIb and HCIId/IIa were recorded only after an 8-day stimulation period. This delay at the protein level was interpreted to relate to the slow turnover of HCIIb which was estimated from its decay in long-term stimulated muscles with an approximate value of 14.7 days. Therefore, protein degradation seems to be an important post-translational regulatory step in the remodeling process of the thick filament.

A simplified method for the determination of nitric oxide in biological solutions.

A new simplified procedure for determination of nitric oxide (NO) in biological solutions is described utilizing a new reducing system of nitric oxide prior to chemiluminescence. Advantages of the new method makes heating of the reducing solution unnecessary and avoids cooling and condensation of generated vapors. Only traces of acid with a high boiling point are used. The method permits analysis of small sample volumes (200 microL). The basal production of nitric oxide by freshly harvested endothelial cells ranged from 100 to 880 picomoles.

Myosin heavy-chain-based isomyosins in developing, adult fast-twitch and slow-twitch muscles.

A modified method of electrophoresis under nondenaturing conditions made it possible to separate rat muscle extracts of defined myosin heavy chain (HC) and light chain (LC) composition into subsets of developmental, fast and slow myosin heavy-chain-based isomyosins. The fastest migrating isomyosins were the neonatal isomyosins (nM1, nM2, nM3), followed by the slightly slower migrating embryonic isomyosins (eM1, eM2, eM3, eM4). Of the nine adult fast isomyosins, the HCIIb-based isomyosins (FM1b, FM2b, FM3b) were the fastest migrating. These were followed by the HCIId-based isomyosins (FM1d, FM2d, FM3d). The HCIIa-based isomyosins (FM1a, FM2a, FM3a) were the slowest. Our results suggest that FM3a is identical with the so-called intermediate isomyosin (IM) described in the literature. The slow myosin heavy-chain-based isomyosins (SM1, SM2, SM3) migrated far behind the fast isomyosins. Whereas the gross electrophoretic mobilities of each of these isomyosin triplets is determined by the specific heavy chain complement, the different mobilities of the bands within each triplet result from different alkali light chain combinations. Thus, the fastest triplet bands of the neonatal (nM1) and adult fast isomyosins (FM1b, FM1d, FM1a) represent the LC3f homodimers, the slowest (nM3, FM3b, FM3d, FM3a) the LC1f homodimers, and the intermediate bands (nM2, FM2b, FM2d, FM2a) the LC1f/LC3f heterodimers. Different proportions of the adult fast isomyosin triplet bands indicate that the affinity for LC3f decreases in the order HCIIb, HCIId, HCIIa. The three slow isomyosins represent LC1sa (SM1) and LC1sb (SM3) homodimers and a LC1sa/LC1sb heterodimer (SM2). Circumstantial evidence suggests an inverse order in rabbit muscle where SM1 and SM3 most likely represent LC1sb and LC1sa homodimers, respectively.

Electrophoretic separation by an improved method of fast myosin HCIIb-, HCIId-, and HCIIa-based isomyosins with specific alkali light chain combinations.

An improved method of electrophoresis under nondenaturing conditions separated three electrophoretically distinct isomyosin triplets when applied to rat fast-twitch muscles displaying a predominance of one of the fast myosin heavy chain isoforms HCIIb, HCIId or HCIIa. The three isomyosin triplets, named FM1b-FM3b, FM1d-FM3d, FM1a-FM3a, corresponded to the three possible alkali light chain (LC) combinations (LC1f homodimer, LC1f/LC3f heterodimer, and LC3f homodimer) with each fast HC isoform. Different proportions of these various isomyosins suggested specific affinities of light chains LC1f and LC3f for the fast heavy chain isoforms.

Antagonistic effects of chronic low frequency stimulation and thyroid hormone on myosin expression in rat fast-twitch muscle.

This study investigates effects of chronic low frequency stimulation (CLFS) on myosin heavy (MHC) and light chain (MLC) expression in fast-twitch muscles in hypothyroid, euthyroid, and hyperthyroid rats. The changes at both the mRNA and protein level indicated antagonistic effects of thyroid hormone and CLFS: under euthyroid conditions, CLFS mainly elicited a MHCIIb----MCHIId----MHCIIa transition. Whereas CLFS did not induce the slow MHCI in the euthyroid state, this isoform was present in the hypothyroid state and was further enhanced with CLFS indicating the suppressive effect of thyroid hormone to be stronger than the inductive influence of CLFS. Hyperthyroidism alone suppressed the expression MHCIIa and enhanced a MHCIId to MHCIIb transition. This shift to the faster MHC isoforms was only partially counteracted by CLFS. Thus, it appeared that thyroid hormone had a graded suppressive effect on the expression of MHC isoforms in the order MHCIId less than MHCIIa less than MHCI. Elevated neuromuscular activity partially counteracted these hormone effects. Changes in MLC mRNAs were consistent with those in the MHC pattern, i.e. increases or decreases in MHCIIb led to corresponding changes in the expression of MLC3f. A similar relationship existed for the slow MHCI and the slow MLC isoforms.

Chronic stimulation-induced effects point to a coordinated expression of carbonic anhydrase III and slow myosin heavy chain in skeletal muscle.

Chronic low-frequency stimulation of rat fast-twitch muscle induces 3.7-fold elevations in cytochrome c oxidase activity, but remains without effect on carbonic anhydrase III (CAIII) mRNA and protein. This is in contrast with the situation in the rabbit where chronic stimulation elicits more than 10-fold elevations in CAIII activity and mRNA content which coincide with an enhanced expression of the slow myosin heavy chain (HCI). Since chronic stimulation of rat muscle does not enhance the expression of HCI, we conclude that CAIII is expressed in parallel with HCI and, therefore, is present only in type I and C fibers.

Changes in myosin heavy chain isoforms during chronic low-frequency stimulation of rat fast hindlimb muscles. A single-fiber study.

Fast-twitch rat muscles contain three fast myosin heavy chains (HC) which can be separated by density gradient gel electrophoresis. Their mobility increases in the order of HCIIa less than HCIId less than HCIIb. In contrast to the rabbit, where chronic low-frequency nerve stimulation induces a fast-to-slow conversion, stimulation for up to 56 days does not lead to appreciable increases in the relative concentration of the slow myosin heavy chain HCI in rat fast-twitch muscles. However, chronic stimulation of rat fast-twitch muscle does evoke a rearrangement of the fast myosin heavy chain isoform pattern with a progressive decrease in HCIIb and progressive increases in HCIIa and HCIId. As judged from the time course and extent of these transitions, it appears that HCIId is an intermediate form between HCIIb and HCIIa. Single-fiber analyses of normal muscles make it possible to assign these heavy chain isoforms to histochemically defined fiber types IIB, IID, and IIA. The stimulation-induced fiber transformations produce numerous hybrid fibers displaying more than one myosin heavy chain isoform. Some transforming fibers contain up to four different myosin heavy chain isoforms.

Myosin heavy chain isoforms in histochemically defined fiber types of rat muscle.

Combined histochemical and biochemical analyses were performed on rat skeletal muscles in order to determine the myosin heavy chain patterns in specific fiber types. Four myosin heavy chain isoforms were separated by gradient polyacrylamide gel electrophoresis of extracts from single fibers and whole muscle homogenates. Their electrophoretic mobility increased in the order HCIIa, HCIIb, and HCI. HCIIa, HCIIb and HCI were present as unique isoforms in histochemically defined fiber types IIA, IIB and I, respectively. The isoforms HCI and HCIIa coexisted at variable ratios in type IC and IIC fibers. An additional fast myosin heavy chain isoform with an electrophoretic mobility between HCIIa and HCIIb was designated as HCIId because of its abundance in fast fibers of large diameter in the diaphragm. With the exception of slight differences in mATPase staining intensity after acid preincubation, these fibers were almost indistinguishable from type IIB fibers. In view of their specific myosin heavy chain composition (HCIId), these fibers were named type IID. In the extensor digitorum longus muscle, type IID fibers were of smaller size than type IIB and differed from the latter by higher NADH tetrazolium reductase activities. Circumstantial evidence suggests that type IID fibers are identical with the 2X fibers, previously described by Schiaffino et al. (1986).