Effects of posture on cardiovascular responses to lower body positive pressure at rest and during dynamic exercise

We tested thehypothesis that cardiovascular responses to lower body positivepressure (LBPP) would be dependent on the posture of the subject andalso on the background condition (rest or exercise). We measured heartrate (HR), mean arterial blood pressure (MAP), and cardiac strokevolume in eight subjects at rest and during cycle ergometer exercise(76 ± 3 W) with and without LBPP (25, 50, and 75 mmHg) inthe supine and upright positions. At rest, the increase in MAP wasproportional to the increase in LBPP and was greater in the supine (6 ± 2, 15 ± 3, and 26 ± 3 mmHg) than in the upright (2 ± 3, 9 ± 3, and 17 ± 3 mmHg) position. During dynamic exercise,the increases in MAP evoked by 25, 50, and 75 mmHg LBPP were greater inthe supine (13 ± 2, 28 ± 3, and 40 ± 3 mmHg) than in theupright (7 ± 3, 12 ± 3, and 25 ± 3 mmHg)position. We conclude that the systemic pressure response to LBPP isclearly dependent on the body position, with the larger pressureresponses being associated with the supine position both at rest andduring dynamic leg exercise.

     

Histopathology of Chlamydia trachomatis salpingitis after primary and repeated reinfections in the monkey subcutaneous pocket model

Monkeys with subcutaneously autotransplanted salpingeal fimbrial tissues were subjected to primary and repeated infections with Chlamydia trachomatis. The inflammatory response after primary inoculation was characterized by infiltration with polymorphonuclear leucocytes in the acute phase and mononuclear cells in the chronic phase. However, the inflammatory response after repeated infections was dominated by a mononuclear cell infiltration with a conspicuous absence of the initial phase of polymorphonuclear leucocyte infiltration. The remarkable findings of repeated infections were plasma cell infiltration, lymphoid follicle formation, and increased fibroblast activity resulting in extensive fibrosis. These findings are similar to those described for monkeys inoculated directly into the oviducts with C. trachomatis and support our original hypothesis that, after chlamydial infection, the tissue damage is provoked by immune-mediated mechanisms.     

Multiple solution conformations and internal rotations of the decapeptide gramicidin S.

The conformations of every C alpha H-C beta H2 moiety of the peptide gramicidin S are reported. Internal rotation occurs, but distinct preferences for one side chain rotamer, greater than 80%, are found for the D-phenylalanine and ornithine residues. Leucine and valine exhibit more extensive averaging while proline is shown to be at least 90% in the Ramachandran B conformation. The data are consistent with the coexistence of many tertiary conformations of gramicidin S; the statistical weights of the twelve major tertiary conformations consistent with the rotamer populations are reported. The relative statistical weights of the tertiary conformers depend upon temperature and solvent. A comparison of the conclusions from this publication and conformations derived by energy minimization procedures is made. Partial agreement was found, but the calculations have not yet predicted the wealth of coexisting tertiary conformations nor accounted for the subtle effects of solvent. It is proposed that a more complete picture of the conformational dynamics of gramicidin S and other peptides will result from calculations which use as a basis the extensive data reported here.     

Chemical trapping of complexes of dihydroxyacetone phosphate with muscle fructose-1,6-bisphosphate aldolase

Dihydroxyacetone phosphate (DHAP) in equilibrium with FDP aldolase of muscle is present in the form of two major covalent complexes. One, representing approximately 60% of total bound substrate, decomposes to Pi and methylglyoxal upon acid denaturation of the enzyme as first reported by Grazi and Trombetta [Grazi, E., & Trombetta, G. (1979) Biochem. J. 175, 361-365]. This is now shown to be the enzyme-eneamine phosphate reaction intermediate since Pi formation is prevented if the acid denaturation is done in the presence of potassium ferricyanide, an oxidant of the eneamine. The enzyme-eneamine aldehyde X Pi 6, presumed to be an intermediate of the slow methylglyoxal synthetase reaction of aldolase, must not be a significant source of the Pi produced upon denaturation and is probably not a significant component of the equilibrium. The oxidation product, the enzyme-imine of phosphopyruvaldehyde, is sufficiently stable in 1 N HCl, t1/2 = 76 min at 0 degree C, to be isolated with the trichloroacetic acid precipitated protein. A second covalent complex, approximately 20-24% of bound dihydroxyacetone [32P]phosphate, remains with the protein during acid denaturation and centrifugation. This acid-stable complex is formed rapidly and is chased rapidly by unlabeled substrate. Its stability in 1 N HCl is similar to that of the ferricyanide-oxidized derivative mentioned above. From this and its reactivity with cyanoborohydride in acid, this complex is thought to be the imine adduct of DHAP with aldolase 4 and/or the carbinolamine complex 3 present in the initial equilibrium. D-Glyceraldehyde 3-phosphate in the carbonyl form also forms an acid-precipitable complex with aldolase.(ABSTRACT TRUNCATED AT 250 WORDS)     

Recovery of staphylococcal enterotoxin from foods by affinity chromatography.

Extraction, concentration, and serological detection of staphylococcal enterotoxins from foods are laborious and time consuming. By exposing food extracts to an insoluble matrix tagged with specific anti-enterotoxin B, we have been able to recover the toxin from foods in a sensitive and rapid way. After mixing the reagents for 2 h at room temperature, immunoglobulin G antibodies were attached to CNBr-activated Sepharose 4B at pH 8.5 (0.1 M carbonate buffer with 0.5 M NaCl). Sepharose-antibody complex (1 ml) specifically recovered 0.1 to 30 mug of enterotoxin B from 400 ml of food extract (100 g of food) after mixing for 2 h at 4 C. The Sepharose-antibody-toxin complex was washed with 0.02 M phosphate-buffered saline at pH 7.2, and the toxin was dissociated by 2 to 4 ml of 0.2 M HCl-glycine plus 0.5 M NaCl buffer at pH 2.8. The recovered enterotoxin was free of interfering food components and could be detected serologically. Work to couple antibodies A, B, C, D, and E to Sepharose to recover all five toxins in one step is under study.     

Antileukemic agent alkyllysophospholipid regulates phosphorylation of distinct proteins in HL60 and K562 cells and differentiation of HL60 cells promoted by phorbol ester

The tumor-promoting 12-0-tetradecanoylphorbol-13-acetate (TPA) stimulated phosphorylation of several proteins in block I (including protein Ia) and protein 3 in HL60 cells. The antileukemic agent alkyllysophospholipid (ALP) inhibited the TPA-stimulated phosphorylation of these proteins and the TPA-induced differentiation of the cells. In comparison, TPA only stimulated phosphorylation of protein 3 in K562 cells which, in contrast, were not induced to differentiate by TPA and lacked protein Ia and had a very high basal phosphorylation of protein B. ALP inhibited phosphorylation of protein 3 as well as protein B in K562 cells. The data suggest that the presence of distinct phosphoproteins and regulation of their phosphorylation may be related to the selective susceptibility of the two leukemia cell lines to the maturating effect of TPA and cytotoxicity of ALP.