Sequence similarity between alpha 2-macroglobulin from the horseshoe crab, Limulus polyphemus, and proteins of the alpha 2-macroglobulin family from mammals.

1. Purified alpha 2-macroglobulin (alpha 2M) from the American horseshoe crab, Limulus polyphemus was cleaved with trypsin and 20 of the tryptic peptides were sequenced and compared with the sequences of human alpha 2M, rat alpha 1M, alpha 2M, and alpha 1-inhibitor 3, and human complement proteins C3 and C4. 2. Ten of the peptides (233 residues), including that containing the thiol ester site, could be aligned unambiguously with stretches in mammalian alpha 2M, with a degree of identity greater than 30%. 3. The 12-residue thiol ester-containing peptide of Limulus alpha 2M showed 67% identity with the same stretch of human alpha 2M.     

Longitudinal changes in the kinetic response to heavy-intensity exercise in children.

The purpose of this study was to investigate longitudinal changes with age in the kinetic response to cycling at heavy-intensity exercise in boys and girls. Twenty-two prepubertal children (13 male, 9 female) carried out a series of exercise tests on two test occasions with a 2-yr interval. On each test occasion, the subject completed multiple transitions from baseline to 40% of the difference between their previously determined V-slope and peak O(2) uptake (Vo(2)) for 9 min on an electronically braked cycle ergometer. Each subject's breath-by-breath responses were interpolated to 1-s intervals, time aligned, and averaged. The data after phase 1 were fit with 1) a double exponential model and 2) a single exponential model within a fitting window that was previously identified to exclude the slow component. There were no significant differences in the parameters of the primary component between each model. Subsequent analysis was carried out using model 2. The Vo(2) slow component was computed as the difference between the amplitude of the primary component and the end-exercise Vo(2) and was expressed as the percent contribution to the total change in Vo(2). Over the 2-yr period, the primary time constant (boys 16.8 +/- 5.3 and 21.7 +/- 5.3 s, girls 21.1 +/- 8.1 and 26.4 +/- 8.4 s, first and second occasion, respectively) and the relative amplitude of the slow component (boys 9.4 +/- 4.6 and 13.8 +/- 5.3%, girls 10.3 +/- 2.4 and 15.5 +/- 2.8%, first and second occasion, respectively) significantly increased with no sex differences. The data demonstrate that children do display a slow-component response to exercise and are consistent with an age-dependent change in the muscles' potential for O(2) utilization.     

Evidence for the Occurrence of Specific Iron (III)-Binding Compounds in Near-Shore Marine Ecosystems

Aqueous extracts of samples from representative types of near-shore marine ecosystems were examined for growth-promoting activity for the siderochrome auxotroph, Arthrobacter JG-9. Blue-green algal mat and sea grass samples gave strongly positive responses. Relative to Desferal (Ciba Pharmaceutical Co., Summit, N.J.), concentrations up to 110 ng/g (wet weight) of sample were found. In contrast, extracts of anaerobic muds or macroalgae gave no detectable activity. The hydroxamate type of iron chelator may be important in providing a form of iron in sea water readily utilized by the microalgae and marine higher plants.     

Effects of part sequences of human growth hormone on in vivo hepatic glycogen metabolism in the rat

Acute effects of two part sequences of human growth hormone on the in vivo activity levels of hepatic glycogen synthase and glycogen phosphorylase were examined. The peptide corresponding to residues 6 to 13 of the hormone (hGH 6–13) decreased the percentage of phosphorylase in the active form without affecting synthase activity. This action was indirect and dependent upon insulin. The peptide hGH 177–191 decreased the level of the active form of synthase without affecting phosphorylase activity. This effect was also observed with analogous peptides containing the sequence hGH 178–191 (i.e., hGH 172–191 and hGH 178–191), whereas the peptide hGH 179–191 was inert.The onset of these effects was rapid, and maximum changes in activity were produced in 5 min by both peptides. The effect for hGH 177–191 was short-lived, and synthase activity had returned to normal levels by 15 min, whereas the action of hGH 6–13 was of longer duration and was still quite marked at 60 min. Both peptides showed a linear dependence of response to the log dose of peptide injected over the range 0.1–250 μg hGH 6–13 per kg body weight and 0.05–25 gmg hGH 177–191 per kg body weight. Hepatic 3′,5′-cyclicadenylic acid levels were not affected by either peptide. Incorporation of glycerol carbon liver glycogen was increased by hGH 6–13 and decreased by hGH 177–191. This discussed in terms of a futile cycle between glycogen and hexone phosphate in the liver, as the basis for a control mechanism for hepatic glycogen metabolism. The present observations are consistent with other in vivo and in vitro actions of these and related peptides.     

A new solid-state microelectrode for measuring intracellular chloride activities.

Solid-state microelectrodes from measuring intracellular Cl activity (alphaiCl) were made by sealing the tips of tapered glass capillaries (tip diameter 0.3 mum), coating them under vacuum with a 0.2-0.3 mum thick layer of spectrscopic grade silver, and sealing them (except for the terminal 2-5 mum of the tip) inside tapered glass shields. 106 microelectrodes had an average slope of 55.0+/- 0.6 m V (S,E,) per decade c hange in alphaCl. Tip resistance was (77.1+/- 3.1) x 10(9) omega(n=30). Electrode response was rapid (10-20 s), was unaffected by HCO3, H2PO4, HPO42 or protein, and remained essentially unchanged over a 24-h period. AlphaiCl in frog sartorius muscle fibers and epithelial cells of bullfrog small intestine was measured in vitro. In both tissues, alphaiCl significantly exceeded the value corresponding to equlibrium ditribution of Cl across the cell membrane.     

Insulin enhances luteinizing hormone-stimulated steroidogenesis by porcine theca cells

It has been shown recently that insulin enhances differentiation of rat, pig, and human granulosa cells. The present studies were done to determine if insulin also plays a role in the regulation of theca cell steroidogenesis. Theca cells were obtained from prepubertal gilts and cultured under serum-free conditions for 48 h. Theca cell androstenedione production under basal and luteinizing hormone (LH)-stimulated conditions was significantly increased by adding insulin (1 microgram/ml) to the culture medium. Treatment of basal and LH-stimulated cultures with increasing concentrations of insulin (0.001-10 micrograms/ml) caused dose- and time-dependent increments in androstenedione production, but the effect was independent of the dose of LH employed. The ability of insulin to enhance thecal cell androstenedione production was mimicked by somatomedin C, but not by relaxin. Studies to determine the mechanism(s) of action of insulin showed that insulin action is exerted, at least in part, at a site(s) proximal to cyclic adenosine 3'5'-monophosphate (cAMP) generation, since insulin enhanced both basal and LH-stimulated accumulation of extracellular cAMP in addition to increasing androstenedione production. This effect was further enhanced by 3-isobutyl-1-methyl xanthine, an inhibitor of phosphodiesterase activity. Insulin treatment also caused dose-dependent increments in forskolin- and prostaglandin E2-stimulated accumulation of extracellular cAMP and androstenedione. Insulin also increased both the basal and LH-stimulated production of progesterone and its precursor pregnenolone, in addition to the increases in androstenedione.(ABSTRACT TRUNCATED AT 250 WORDS)