The effects of interelectrode distance on electromyographic amplitude and mean power frequency during isokinetic and isometric muscle actions of the biceps brachii.

The purpose of this study was to examine the effects of interelectrode distance (IED) on the absolute and normalized electromyographic (EMG) amplitude and mean power frequency (MPF) versus isokinetic and isometric torque relationships for the biceps brachii muscle. Ten adults [mean+/-SD age=22.0+/-3.4 years] performed submaximal to maximal, isokinetic and isometric muscle actions of the dominant forearm flexors. Following determination of isokinetic peak torque (PT) and the isometric maximum voluntary contraction (MVC), the subjects performed randomly ordered, submaximal step muscle actions in 10% increments from 10% to 90% PT and MVC. Surface EMG signals were recorded simultaneously from bipolar electrode arrangements placed over the biceps brachii muscle with IEDs of 20, 40, and 60mm. Absolute and normalized EMG amplitude (muVrms and %max) increased linearly with torque during the isokinetic and isometric muscle actions (r(2) range=0.988-0.998), but there were no significant changes for absolute or normalized EMG MPF (Hz or %max) from 10% to 100% PT and MVC. In some cases, there were significant (p<0.05) differences among the three IED arrangements for absolute EMG amplitude and MPF values, but not for the normalized values. These findings suggested that for the biceps brachii muscle, IEDs between 20 and 60mm resulted in similar patterns for the EMG amplitude or MPF versus dynamic and isometric torque relationships. Furthermore, unlike the absolute EMG amplitude and MPF values, the normalized EMG data were not influenced by changes in IED between 20 and 60mm. Thus, normalized EMG data can be compared among previous studies that have utilized different IED arrangements.     

Lost in translation: The search for an in vitro screen for spermatogenic toxicity

The last two decades have seen an increasing search for in vitro models that can replace the use of animals for safety testing. We adapted the methods from a recent nonquantitative report of spermatogenesis occurring in ex vivo mouse testis explants and tried to develop them into a screening assay. The model consisted of small pieces of neonatal mouse testis (testis “chunks”), explanted and placed on pillars of agarose or chamber inserts, and cultured at the air–liquid interface. A peripheral torus‐shaped zone in these explants would often contain tubules showing spermatogenesis, while the middle of each chunk was often necrotic, depending on the thickness of the tissue. The endpoint was histology: what proportion of tubules in the “permissive torus” actually contained healthy pachytene spermatocytes or spermatids? Extensive statistical modeling revealed that a useful predictive model required more than 60% of these tubules to show spermatogenesis. Separately, the logistics of running this as a predictive assay require that the controls consistently produce ≥ 60% tubules with pachytenes and round spermatids, and achieving this level of spermatogenesis reliably and consistently every week proved ultimately not possible. Extensive trials with various media additions and amendments proved incapable of maintaining the frequency of spermatogenic tubules at consistently ≥ 60%. Congruent with Schooler's “decline effect”; generally, the more often we ran these cultures, the worse the performance became. We hope that future efforts in this area may use our experience as a starting point on the way to a fully productive in vitro model of spermatogenesis.     

Effect of incubation of guinea-pig taenia coli in potassium-free media on arachidonate release and lipid metabolism

We studied the effects of immersion of guinea-pig taenia coli strips in potassium-free media on arachidonate stores and other lipid fractions. Control studies obtained with the strips in Krebs solution showed that greater than 97% of arachidonate was found esterified in phospholipid with the following distribution: phosphatidylethanolamine greater than phosphatidylcholine greater than phosphatidylserine plus phosphatidylinositol. 30 min incubation of the strips with [3H]arachidonate complexed to albumin resulted in incorporation of this isotope into phospholipid and neutral lipid fractions, phosphatidylcholine greater than neutral lipid greater than phosphatidylserine plus phosphatidylinositol greater than phosphatidylethanolamine. 30 min incubations with 32PO4(2-)-resulted in an isotope incorporation into phospholipids, phosphatidylcholine greater than phosphatidylserine plus phosphatidylinositol greater than phosphatidylethanolamine. After 'loading' with [3H]arachidonate and 32P, placing the strips in potassium-free media caused the following: there was an increased release of [3H]arachidonate from the tissue into the bathing solution. [3H]Arachidonate and 32P radioactivity in phosphatidylinositol fell without a change in phosphatidylinositol content. [3H]Arachidonate and 32P radioactivity in other phospholipid fractions was unchanged. Arachidonate specific activity fell and arachidonate content increased in the phosphatidylserine plus phosphatidylinositol fraction. [3]Arachidonate in neutral lipid did not change significantly. We conclude that exposure of taenia coli to potassium-free media activates turnover of phosphatidylinositol, which results in release of arachidonate.     

Attractiveness of the male Acheta domesticus calling song to females

1. Most crickets first demonstrated positive phonotaxis to 65 dB CSs having a 53-62 ms SP by day 3 following the imaginal molt (Fig. 3B). The onset of copulatory readiness occurred on average at 3.2 days. 2. The attractive range of SPs for most females became progressively broader as they aged (Fig. 4). Three to 4-day-old females were attracted to a smaller number of CS SPs than were 20-21 day old females (Fig. 4). 3. Older, less selective females did not typically respond to the same range of CS SPs (Fig. 6). However, they were more likely to respond to some SPs (especially 50 ms) than to others (Fig. 7). 4. The phonotactic threshold decreased from 95 dB or greater on day 0 to a mean of 55 dB by day 3, during a period of increasing JHIII biosynthesis, and thereafter remained at that level (Fig. 8). 5. During a period of maximal JHIII production, 3-5 day-old females usually responded to 4 of the 7 SPs presented (Fig. 8). Females older than 12 days were unselective for CS SP, and JHIII synthesis remained at a level below the peak production on day 4 (Fig. 8). 6. Older females, that were unselective for CS SP, became as selective as 3 to 5-day-old females within 4 days of topical application of JHIII (Figs. 9-11).     

ADP-ribosylation by cholera toxin: functional analysis of a cellular system that stimulates the enzymic activity of cholera toxin fragment A1

We have clarified relationships between cholera toxin, cholera toxin substrates, a membrane protein S that is required for toxin activity, and a soluble protein CF that is needed for the function of S. The toxin has little intrinsic ability to catalyze ADP-ribosylations unless it encounters the active form of the S protein, which is S liganded to GTP or to a GTP analogue. In the presence of CF, S.GTP forms readily, though reversibly, but a more permanent active species, S-guanosine 5'-O-(3-thiotriphosphate) (S.GTP gamma S), forms over a period of 10-15 min at 37 degrees C. Both guanosine 5'-O-(2-thiodiphosphate) and GTP block this quasi-permanent activation. Some S.GTP gamma S forms in membranes that are exposed to CF alone and then to GTP gamma S, with a wash in between, and it is possible that CF facilitates a G nucleotide exchange. S.GTP gamma S dissolved by nonionic detergents persists in solution and can be used to support the ADP-ribosylation of nucleotide-free substrates. In this circumstance, added guanyl nucleotides have no further effect. This active form of S is unstable, especially when heated, but the thermal inactivation above 45 degrees C is decreased by GTP gamma S. Active S is required equally for the ADP-ribosylation of all of cholera toxin's protein substrates, regardless of whether they bind GTP or not. We suggest that active S interacts directly with the enzymic A1 fragment of cholera toxin and not with any toxin substrate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Identification of pyridoxine 3-sulfate, pyridoxal 3-sulfate, and N-methylpyridoxine as major urinary metabolites of vitamin B6 in domestic cats

Preliminary studies of vitamin B6 metabolism in three adult domestic cats detected very little pyridoxic acid in the urine. At oral doses of 49 to 490 mumol of [14C]pyridoxine hydrochloride, 50% of the excreted dose occurred as pyridoxine 3-sulfate and 25% as N-methylpyridoxine. The identity of these two metabolites was confirmed by isolation from urine and comparison with known compounds. A third compound was identified as pyridoxal 3-sulfate on the basis of chromatographic behavior and fluorescent properties before and after hydrolysis. At pyridoxine intakes of 0.97 mumol/day, the concentration of pyridoxal 3-sulfate in the urine sometimes exceeded the concentration of pyridoxine 3-sulfate. Pyridoxic acid remained a minor urinary metabolite at pyridoxine intakes ranging from 0.97 to 490 mumol/day. Although sulfation of phenol groups and methylation of the ring nitrogen are well-known detoxication reactions, this appears to be the first time such reactions have been observed in normal metabolism of vitamin B6. These observations provide further evidence of the diversity of vitamin B6 metabolism between species. While such diversity complicates the extrapolation of data from animal studies to humans, it does provide a variety of models for examining the influences of various factors on vitamin B6 metabolism.     

Effect of four different starting stances on sprint time in collegiate volleyball players

Starting stance plays an important role in influencing short-distance sprint speed and, therefore, the ability to reach a ball during sport play. The purpose of this study was to evaluate 4 different starting stances on sprint time. Twenty-six male and female collegiate volleyball players volunteered to participate in 1 testing session. Each subject performed 3 15-ft sprint trials at each of 4 different starting stances (P-parallel, FS-false step, S-staggered, and SFS-staggered false step) in random order. Analysis of variance revealed that there was no significant interaction of sex by stance, but there were main effects for sex (men were faster than women) and stance. The FS (1.18 ± 0.10 seconds), S (1.16 ± 0.07 seconds), and SFS (1.14 ± 0.06 seconds) stances were faster than the P (1.25 ± 0.09 seconds) stance, and the SFS stance was faster than the FS stance. This indicates that starting with a staggered stance (regardless of stepping back) produced the greatest sprinting velocity over the initial 15 feet. Although taking a staggered stance seems counterproductive, the resultant stretch-shortening cycle action and forward body lean likely increase force production of the push-off phase and place the total body center of mass ahead of the contacting foot, thereby, decreasing sprint time.     

Responses of human endothelial cells to pathogenic and non-pathogenic Leptospira species

Leptospirosis is a widespread zoonotic infection that primarily affects residents of tropical regions, but causes infections in animals and humans in temperate regions as well. The agents of leptospirosis comprise several members of the genus Leptospira, which also includes non-pathogenic, saprophytic species. Leptospirosis can vary in severity from a mild, non-specific illness to severe disease that includes multi-organ failure and widespread endothelial damage and hemorrhage. To begin to investigate how pathogenic leptospires affect endothelial cells, we compared the responses of two endothelial cell lines to infection by pathogenic versus non-pathogenic leptospires. Microarray analyses suggested that pathogenic L. interrogans and non-pathogenic L. biflexa triggered changes in expression of genes whose products are involved in cellular architecture and interactions with the matrix, but that the changes were in opposite directions, with infection by L. biflexa primarily predicted to increase or maintain cell layer integrity, while L. interrogans lead primarily to changes predicted to disrupt cell layer integrity. Neither bacterial strain caused necrosis or apoptosis of the cells even after prolonged incubation. The pathogenic L. interrogans, however, did result in significant disruption of endothelial cell layers as assessed by microscopy and the ability of the bacteria to cross the cell layers. This disruption of endothelial layer integrity was abrogated by addition of the endothelial protective drug lisinopril at physiologically relevant concentrations. These results suggest that, through adhesion of L. interrogans to endothelial cells, the bacteria may disrupt endothelial barrier function, promoting dissemination of the bacteria and contributing to severe disease manifestations. In addition, supplementing antibiotic therapy with lisinopril or derivatives with endothelial protective activities may decrease the severity of leptospirosis.