The pro-protein convertase PC1 is induced in the transected sciatic nerve and is present in cultured Schwann cells: comparison with PC5, furin and PC7, implication in pro-BDNF processing

Emotional dissonance, or person-role conflict originating from the conflict between expressed and experienced emotions, was examined. The study was based on a reconceptualization of the emotional labor construct, with dissonance as a facet rather than a consequence of emotional labor. The effects of emotional dissonance on organizational criteria were isolated, thereby explaining some of the conflicting results of earlier studies. Empirically, job autonomy and negative affectivity as antecedents of emotional dissonance, and emotional exhaustion and job satisfaction as consequences of emotional dissonance, were explored. Self-monitoring and social support were tested as moderators of the emotional dissonance-job satisfaction relationship. Significant relationships with job autonomy, emotional exhaustion, and job satisfaction were found. Social support significantly moderated the emotional dissonance-job satisfaction relationship.     

Nerve growth factor promotes activation of the alpha, beta and gamma isoforms of protein kinase B in PC12 pheochromocytoma cells

The activation of phosphatidylinositol (PtdIns) 3-kinase is considered to be a key event occurring after stimulation of cells with growth factors. The proto-oncogenic protein kinase B (PKB; also known as RAC protein kinase or Akt) has recently been shown to be a downstream target of PtdIns 3-kinase and may be involved in cell survival. We therefore asked whether stimulation of neuronal cells with nerve growth factor (NGF), on which certain types of neurons are dependent for survival, causes activation of PKB. Stimulation of serum-starved PC12 rat pheochromocytoma cells with NGF caused an increase of up to 14-fold in PKB activity. This activation was detected within 1 min of stimulation and occurred at NGF concentrations that are consistent with TrkA-mediated signaling. PKB activation was accompanied by a decrease in electrophoretic mobility of the kinase, which is characteristic of phosphorylation. Both PKB activation and mobility changes were prevented by wortmannin, indicating the upstream involvement of PtdIns 3-kinase in these events. Analyses employing isoform-specific antibodies for immunoprecipitation suggested that all three isoforms of PKB (alpha, beta and gamma) are activated in response to NGF. G-protein-coupled-receptor agonists, lysophosphatidic acid (lyso-PtdH) and thrombin, which induce rapid neurite retraction, neither stimulated PKB activity, nor affected NGF-induced or insulin-induced kinase activation. Wortmannin treatment did not prevent neurite retraction induced by lyso-PtdH or thrombin. These data suggest that PtdIns 3-kinase and PKB are not involved in cytoskeletal changes mediated by the small GTPase Rho.     

Effects of growth rate and cell density on nerve growth factor secretion in cultures of vascular and bladder smooth muscle cells from hypertensive and hyperactive rats

Changes in action potential parameters by and inotropic responses to nicardipine, verapamil, ryanodine and cyclopiazonic acid were examined in isolated ventricular myocardial preparations from neonatal and adult mice. The action potential of both neonatal and adult mice had a unique configuration with little evidence of a plateau at depolarized membrane potential; the action potential duration was significantly larger in neonatal preparations. Nicardipine had no effect on action potential parameters in the adult while it significantly shortened the action potential duration at 50% repolarization in the neonate. Ryanodine significantly shortened the action potential duration at 80% repolarization at both ages: the shortening was significantly larger in the adult when compared with the neonate. The contraction of ventricular preparations from adult mice were relatively resistant to nicardipine and verapamil. Nicardipine or verapamil, even at 10(-5) M, only decreased the contractile force to 70% of control values; the decrease was much less than that reported in other experimental species such as chick, guinea pig or rabbit. In the neonate, 10(-5) M nicardipine or verapamil decreased the contractile force to 30% of control values. Ryanodine had a potent negative inotropic effect both in the neonate and adult; the effect was significantly larger in the adult. Cyclopiazonic acid produced a decrease in contractile force and prolongation of the time required for relaxation; both effects were significantly larger in the adult. These results suggest that the contraction of the adult mouse myocardium is highly dependent on SR function and less dependent on transsarcolemmal Ca2+ influx when compared with the myocardium of the neonatal mouse and that of other species.     

Phenotypic alterations of neuropeptide Y, vasoactive intestinal peptide and choline acetyltransferase in rat cultured chromaffin cells as effected by nerve growth factor and glucocorticoid

We assessed changes in neuropeptide Y (NPY), vasoactive intestinal peptide (VIP) and choline acetyltransferase (ChAT) immunoreactivities when neonatal rat chromaffin cells were cultured in a medium containing nerve growth factor (NGF), or the synthetic glucocorticoid dexamethasone (DEX), examining whether their expression was correlated with the morphological changes induced by NGF and DEX. All of the chromaffin cells in culture were tyrosine hydroxylase (TH)-immunopositive regardless of whether they extended processes. TH-immunoreactive materials of NGF-treated chromaffin cells were distributed in all the cytoplasmic processes, even at the tips of growth cones. The percentage of NPY-positive chromaffin cells did not change markedly when treated with NGF or DEX throughout the 14 days in culture. The proportion of VIP-positive chromaffin cells increased gradually in the NGF-treated group and that of ChAT-positive cells in the group was similar to VIP. The morphological alterations induced by NGF were not correlated with the changes in proportions of NPY-, VIP- or ChAT-positive chromaffin cells. The percentages of VIP- or ChAT-immunopositive chromaffin cells in the NGF-treated group showed much greater increases than those in the DEX-treated group. These findings suggest that NGF might modulate the phenotypic changes of neuropeptides and amines in rat chromaffin cells.     

Scanning electron microscopic studies of smooth muscle cells and their collagen fibrillar sheaths in empty, distended and contracted urinary bladders of the guinea pig

By employing various synthetic substrates, as well as soluble denatured protein substrate (TAP-lysozyme) and its derivatives, endopeptidase activity of cathepsin C, dipeptidyl aminopeptidase I [EC 3.4.14.1], from bovine spleen was investigated. Cathepsin C efficiently degraded Z-Phe-Arg-MCA, Pro-Phe-Arg-MCA, and Suc-Leu-Leu-Val-Tyr-MCA. This endopeptidase activity required sulfhydryl reagents and halide ions, as in the case of the dipeptidyl aminopeptidase (DAP) activity. We confirmed that this endopeptidase activity is due to cathepsin C itself based on the results on gel-filtration and anion-exchange chromatographies, comparative studies of the inhibitory effects of leupeptin and E-64 on this activity and those of cathepsins B and L, and further the competitive inhibitions by mutual substrates for the DAP and endopeptidase activities of cathepsin C. We also found that cathepsin C endopeptidase activity towards TAP-lysozyme and its N-alpha-acetylated tryptic peptides showed marked dependence on sulfhydryl reagents and chloride ion. Thus, we concluded that cathepsin C has endopeptidase activity as well as DAP activity. The binding energy between the enzyme and the amino acid side chains of the substrate may be as important for the endopeptidase activity as is the electrostatic interaction between the enzyme and the free alpha-amino group of the substrate for the DAP activity.