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The effects of the phorbol ester 12-O-tetradecanoyl phorbol 13-acetate (TPA), an activator of protein kinase C (PKC), and the PKC inhibitor staurosporine on GnRH secretion and mRNA levels were studied in GT1-7 hypothalamic neuronal cells. Dose-response and time-course studies revealed that TPA (10(-8) M) acutely increased GnRH secretion 3-fold at 3-6 h, which then declined to baseline at 24 h, while it progressively decreased GnRH mRNA levels by 50% and 70% at 6 and 24 h, respectively. To ensure that these effects were due to activation and not down-regulation of PKC, cells were treated for 30 min with TPA (10(-8) M). This brief exposure to TPA also resulted in a decrease (60%) in GnRH mRNA levels at 6 h, with a 1.5- to 2-fold increase in GnRH secretion compared to control values, suggesting that activation of PKC decreases the pretranslational expression of GnRH while increasing GnRH secretion. Additional studies measured PKC activity and documented a shift from a cytosolic to a membrane fraction after incubation with TPA, again supporting PKC activation. Exposure of GT1-7 cells to staurosporine (10(-8) M), a PKC inhibitor, resulted in no change in the level of GnRH mRNA or secretion at 6 h. However, incubation with both TPA and staurosporine prevented the decrease in GnRH mRNA levels and partially blocked the increase in GnRH secretion induced by TPA. We conclude that TPA, by activating the PKC pathway, acutely increases GnRH secretion, but dramatically decreases GnRH gene expression. The exact mechanism of these divergent effects on the synthesis and secretion of GnRH remain to be elucidated. 相似文献
63.
OBJECTIVE: Few studies exist on gait adaptation caused by knee osteoarthritis (OA), and those have only explored adaptations of the kinematics and kinetics of the knee joint itself. We characterize ankle, knee, hip, and low back mechanical energy expenditures (MEE) and compensations (MEC) during gait in patients with knee OA. METHODS: Thirteen elderly patients with unilateral knee OA and 10 matched healthy elderly controls were studied during preferred and paced speed gait. Gait speed, step length, and lower extremity and low back joint MEE and MEC were compared between groups. RESULTS: Patients with knee OA had lower, but not significantly different, walking speed and step length compared to the controls, and had significantly different joint kinetic profiles. Patients had reduced ankle power at terminal stance, lacked a second positive peak in knee power, and had increased power absorption at the hip. Abnormal knee kinematics were exaggerated when walking at a paced speed, but hip kinetics normalized among patients with OA. CONCLUSION: Reduced ankle plantar-flexion power in patients with knee OA was probably due to disrupted transfer of energy through the knee. Lack of concentric knee power supports prior studies' conclusions that patients with knee OA avoid using their quadriceps to stabilize the knee, probably to reduce articular loads. Patients with knee OA increase eccentric hip power due to increased hip extension caused by abnormal knee kinematics, potentially increasing hip articular forces. This passive mechanism, however, may assist in the advancement of the leg into swing phase. 相似文献
64.
E A Kuenzel E G Krebs 《Proceedings of the National Academy of Sciences of the United States of America》1985,82(3):737-741
A synthetic peptide having the sequence Arg-Arg-Arg-Glu-Glu-Thr-Glu-Glu-Glu was found to serve as a convenient substrate for the protein kinase generally referred to as casein kinase II. The enzyme exhibited an apparent Km of 500 microM for the peptide, as compared to an apparent Km of 50 microM for casein. The maximum velocities for phosphorylation of the peptide and of casein were similar. The peptide was not phosphorylated by any of eight other protein kinases, all of which were shown to be active toward their known substrates. The peptide was used to monitor activity during steps in the purification of casein kinase II from bovine liver. These experiments demonstrated that with this peptide it is now possible to obtain specific measurements of casein kinase II activity in crude enzyme preparations. 相似文献
65.
Lutz M Rosenberg M Kiessling F Eckstein V Heger T Krebs J Ho AD Katus HA Frey N 《Cardiovascular research》2008,77(1):143-150
AIMS: Recent studies have shown that stem cell therapy may alleviate the detrimental effects of myocardial infarction. Yet, most of these reports observed only modest effects on cardiac function, suggesting that there still is need for improvement before widespread clinical use. One potential approach would be to increase migration of stem cells to the heart. We therefore tested whether local administration of stem cell factor (SCF) improves myocardial homing of intravenously infused lin-/c-kit+ stem cells after myocardial infarction. METHODS AND RESULTS: Myocardial infarction was induced in mice via ligation of the left anterior descending artery and 2.5 microg of SCF were injected into the peri-infarct zone. Sham-operated mice and animals with intramyocardial injection of phosphate-buffered saline (PBS) served as controls. Twenty-four hours after myocardial infarction, lin-/c-kit+ stem cells were separated from murine bone marrow by magnetic cell sorting, labelled with the green fluorescent cell tracker CFDA or 111 Indium, and subsequently 750 000 labelled cells were systemically infused via the tail vein. Another 24 or 72 h later, respectively (i.e. 48 and 96 h after myocardial infarction), hearts were removed and analysed for myocardial homing of stem cells. Green fluorescent stem cells were exclusively detected in the peri-infarct zone of animals having prior SCF treatment. Radioactive measurements revealed that an intramyocardial SCF injection significantly amplified myocardial homing of lin-/c-kit+ stem cells compared to animals with PBS injections (3.58 +/- 0.53 vs. 2.28 +/- 0.23 cpm/mg/10(6)cpm, +60%, P < 0.05) and sham-operated mice without myocardial infarction (3.58 +/- 0.53 vs. 1.95 +/- 0.22 cpm/mg/10(6)cpm, +85%, P < 0.01). Similar results were obtained 72 h after stem cell injection. CONCLUSION: We demonstrate that intramyocardial administration of SCF sustainably directs more lin-/c-kit+ stem cells to the heart. Future studies will have to show whether higher levels of myocardial SCF (i.e. by virus-mediated gene transfer) can further improve homing of systemically delivered c-kit+ stem cells and thus favourably influence cardiac remodelling following myocardial infarction. 相似文献
66.
Meenakshi Hegde Malini Mukherjee Zakaria Grada Antonella Pignata Daniel Landi Shoba A. Navai Amanda Wakefield Kristen Fousek Kevin Bielamowicz Kevin K.H. Chow Vita S. Brawley Tiara T. Byrd Simone Krebs Stephen Gottschalk Winfried S. Wels Matthew L. Baker Gianpietro Dotti Maksim Mamonkin Malcolm K. Brenner Jordan S. Orange Nabil Ahmed 《The Journal of clinical investigation》2021,131(13)
67.
Eva Bundgaard Martin Helgesen Jon E. Carlé Frederik C. Krebs Mikkel Jørgensen 《Macromolecular chemistry and physics.》2013,214(14):1546-1558
The development of new advanced polymers for improving the stability of OPV is reviewed. Two main degradation pathways for the OPV active layer are identified: photochemically initiated reactions primarily starting in the side chains and morphological changes that degrade the important nanostructure. Chemical units can be introduced that impart an increased stability. Similarly, the morphological degradation of the optimal nanostructure can be reduced. Active polymers and blends with acceptor material are used to create nanoparticle links with controlled size. Most of these advanced polymers and processing methods have only been utilized in small‐scale devices prepared by standard techniques such as spin coating, but a few cases of roll‐to‐roll processed solar cells with heat‐cleaved side chains are discussed.
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69.
Synthetic hexapeptide substrates and inhibitors of 3'':5''-cyclic AMP-dependent protein kinase. 总被引:11,自引:6,他引:11 下载免费PDF全文
B E Kemp E Benjamini E G Krebs 《Proceedings of the National Academy of Sciences of the United States of America》1976,73(4):1038-1042
The substrate specificity of the catalytic subunit of rabbit skeletal muscle 3': 5'-cyclic AMP-dependent protein kinase (EC 2.7.1.37; ATP: protein phosphotransferase) has been studied using the synthetic peptide Arg-Gly-Tyr-Ser-Leu-Gly corresponding to the sequence around serine 24, a phosphorylation site in reduced, carboxymethylated, maleylated (RCMM) chicken egg white lysozyme. This peptide served as a substrate for the enzyme and exhibited a 6-fold higher Vmax and a 100-fold higher Km than RCMM-lysozyme. Replacement of the arginine with glycine, histidine, or lysine resulted in a dramatic reduction in the Vmax. These results support the concept that arginine is an important residue in determining the substrate specificity of the protein kinase, predominantly influencing the Vmax of the phosphorylation reaction. Two synthetic peptides in which serine was replaced by an alanine acted as competitive inhibitors of phosphorylation of the synthetic peptide substrate and RCMM-lysozyme. 相似文献
70.