Evidence for the involvement of protein kinase C isoforms in alpha-1 adrenergic activation of phospholipase A2 in FRTL-5 thyroid cells

BACKGROUND: FRTL-5 thyroid cells are a cell line extensively used for the investigation of thyroid functions. Activation of alpha-1 adrenergic receptors stimulates both arachidonic acid (AA) release and cytosolic Ca2+ increase in this cell line. Cytosolic Ca2+ and arachidonic acid are known to be important second messengers regulating a variety of thyroid functions. The generation of these messengers is regulated primarily by two different types of phospholipases, phospholipase C (PLC) and phospholipase A2 (PLA2). METHODS: Norepinephrine (NE, 10 mumol/L) was used as an alpha-1 adrenergic activator, and cytosolic-free Ca2+ concentration ([Ca2+]i) was determined using the fluorescent dye indo-1. Arachidonic acid release was measured as an indicator of PLA2 activation, and protein kinase C (PKC) activity determination and isoforms identification were performed using commercial kits. RESULTS: Norepinephrine increased [Ca2+]i and AA release. Prevention of NE-induced cytosolic Ca2+ influx, either by removal of extracellular Ca2+ or by use of Ca2+ channel blockers, NiCl2 or CoCl2, inhibited AA generation entirely. Inhibition of NE-induced increase in [Ca2+]i by the Ca2+ chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), also significantly suppressed NE-induced AA release. Inhibition of PKC activity by PKC inhibitors (H-7 or staurosporine) or downregulation induced by prolonged treatment with phorbol 12-myristate 13-acetate (PMA) or thyleametoxin (TX) significantly blocked the NE-induced AA release, which indicates PKC is involved in mediating NE-induced AA release. Protein kinase C activity measurement indicated that NE induced an activation of PKC in 5 minutes. To further characterize the role of PKC or Ca2+ in regulation of AA release, we identified PKC isoforms by immunoblotting with specific antibodies against 8 different Protein kinase C isoforms. PKC-alpha, -beta I, -beta II, -gamma, delta, -epsilon, -zeta, and -eta isoforms were identified. Norepinephrine induced translocation of PKC-alpha, -beta I, -beta II, -gamma, -delta, and -epsilon isoforms but not -zeta and -eta from cytosol to membrane. Chelation of intracellular Ca2+, prevention of Ca2+ influx, or prolonged treatment with thymeleatoxin (TX) completely blocked the NE-induced translocation of PKC-alpha. CONCLUSIONS: These results, taken together with data obtained from AA experiments, suggest that PKC plays a critical role in alpha-1 adrenergic receptor mediated PLA2 activation and subsequent AA release. Extracellular Ca2+ influx is a prerequisite for both PKC-alpha translocation and AA release. Whether Ca2+ acts directly upon the PLA2, or via PKC-alpha, to regulate AA generation is an intriguing question that remains to be clarified.     

A 1000 A/20 kV/25 kHz-500 kHz Volt-Ampere-Wattmeter for loads withpower factors from 0.001 to 1.00

Many industrial processes and scientific experiments utilize large amounts of ac power at frequencies from 3 kHz to 500 kHz. The phase angle and the impedances of these loads often vary over a wide range. This paper describes an instrument to provide an accurate measurement of currents (1 A to 1000 A), voltages (100 V to 20 kV), and powers (100 W to 20 MW) over the frequency range from 25 kHz to 500 kHz. It deals with loads having power factors down to nearly zero and with load impedances from 10 Ω to 20 kΩ. The paper contains practical details of both the design and the calibration of the front-end voltage and current transducers. The characteristics of a nearly ideal broadband current transducer are presented. Overall instrument calibration, verification and traceability problems are considered in detail     

New and used temporal models: An issue of time

TheSpecial Issue on Applications of Temporal Models raises many issues of time: What are the important properties of time? How can time be best represented? How can one reason about time-dependent properties? What are the important directions of temporal research? This introductory piece very briefly surveys the current wide variety of temporal models, temporal reasoning methods, and applications to time-varying phenomena. Promising areas of investigation such as the verification of concurrent systems, knowledge-base representation methods, and dealing with theFrame Problem pass in fleeting review. Brief introductions to each of the works in the volume close the section.     

Laser turn-on delay and chirp noise effects in Gb/sintensity-modulated direct-detection systems

This paper describes a new IM/DD optical transmission system simulation which incorporates the impact of laser diode stochastic turn-on effects on system performance. Using this simulation, experimentally observed error rate floors, due to inappropriate laser diode biasing, can be theoretically modelled and predicted, for the first time     

Magnetic resonance staging of neoplasms of the uterus

The treatment of patients with uterine neoplasms may be significantly altered by the stage of disease at the time of diagnosis. A noninvasive and accurate means of staging these tumors is therefore desirable. This article discusses the magnetic resonance imaging techniques and findings that are essential for the accurate staging of uterine neoplasms. The imaging findings are presented following a discussion of the histopathologic findings, clinical presentation, diagnosis and staging, pathways of tumor spread, and treatment of each neoplasm. A comparison of magnetic resonance and other imaging techniques is also provided.