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.     

Muon spin relaxation study of amorphous Hf2Co

Muon spin relaxation measurements were carried out in zero magnetic field on amorphous Hf₂Co. The results obtained and comparison with the behaviour of the muon in crystalline Hf₂Co (A. Baudry, P. Boyer, L.P. Ferreira, S.W. Harris, S. Miraglia and L. Pontonnier, J. Phys.: Condens. Matter, 4 (1992) 5025) indicate that muons occupy interstitial sites without Co atoms in first-neighbour positions and diffuse through those sites, suggesting the existence of some degree of short-range order in the amorphous structure. Muon diffusion cannot be described by a single thermally activated process and is faster in the amorphous alloy than in the crystalline alloy.     

Fatigue behaviour of a phenolic matrix composite

This paper presents results of a fatigue life investigations carried out in plate specimens of a fibre-glass-reinforced phenolic matrix composite. Tensile and Young's modulus data were obtained at four different testing temperatures (room temperature, 100, 150 and 200 °C). The fatigue S−N data were obtained at room temperature only and for two stress ratio values (R=0 and 0.4). Fatigue and tensile behaviour was assesesed in the composite with the fibres aligned in the longitudinal loading direction. The results were obtained for two values of volume fraction (0.28 and 0.42) and three different glass surface treatments. A detailed comparison of fatigue results is given taking into account several fatigue parameters and also the testing variables. Results of observations of SEM fracture surfaces are also presented.     

Analysis and Design of Robust Controllers Using the Interval Diophantine Equation

In this paper the design of pole placement feedback controllers for interval plants is addressed within an Interval Analysis framework. Three main topics connected by the concept of interval Diophantine equation are treated: a relatively straightforward investigation about robust coprimeness of interval polynomials, a reliable computing approach for the design of pole placement controllers in the presence of inaccuracies of interval type, and the design of robust controllers with regional pole placement specifications. After explicitly characterizing a convex subset of robust controllers, the problem of designing non-fragile controllers is formulated as a design centering problem, which is then solved by a global optimization algorithm. Numerical examples illustrate the main characteristics of the approach proposed.This paper has been partially supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico–Brazil.     

Optimal Locations of Groundwater Extractions in Coastal Aquifers

A regional water supply management model for coastal aquifers was developed. One of its outcomes is the definition of the optimized locations for groundwater withdrawal. Such a tool permits the analysis of alternative plans for groundwater extraction and the sustainable use of water resources in a coastal aquifer subject to saltwater intrusion. The principal components are the evolutionary optimization and the analytical/numerical simulation models. The optimization technique looks for the best well locations taking into consideration the economic results and the satisfaction of the societal water demand. However these two concerns are conditioned by trying to control the saltwater intrusion, i.e., preserving the environmental equilibrium. The simulation model uses the governing mathematical equations for groundwater movement to find the interface between freshwater and saltwater. Because of the non-linearity in the system and the possibility of a jumping interface, a security distance was defined. This is a controlling variable which can be set by the decision makers. The model was applied to a typical case with interesting results. For example, diagrams showing the relationship between the location of the wells and the security distance(s) are of importance to the managers. It was also crucial to have an understanding of the tradeoffs between groundwater withdrawals, positions of the wells from the coast line, and the security distance. The model was also applied to a real case in order to relate the extractions, distances and artificial recharge (not presented in this paper).