Submitochondrial distribution of three key steroidogenic proteins (steroidogenic acute regulatory protein and cytochrome p450scc and 3beta-hydroxysteroid dehydrogenase isomerase enzymes) upon stimulation by intracellular calcium in adrenal glomerulosa cells

In adrenal glomerulosa cells, angiotensin II (Ang II) and potassium stimulate aldosterone synthesis through activation of the calcium messenger system. The rate-limiting step in steroidogenesis is the transfer of cholesterol to the inner mitochondrial membrane. This transfer is believed to depend upon the presence of the steroidogenic acute regulatory (StAR) protein. The aim of this study was 1) to examine the effect of changes in cytosolic free calcium concentration and of Ang II on intramitochondrial cholesterol and 2) to study the distribution of StAR protein in submitochondrial fractions during activation by Ca2+ and Ang II. To this end, freshly prepared bovine zona glomerulosa cells were submitted to a high cytosolic Ca2+ clamp (600 nM) or stimulated with Ang II (10 nM) for 2 h. Mitochondria were isolated and subfractionated into outer membranes, inner membranes (IM), and contact sites (CS). Stimulation of intact cells with Ca2+ or Ang II led to a marked, cycloheximide-sensitive increase in cholesterol in CS (to 143 +/- 3. 2 and 151.1 +/- 18.1% of controls, respectively) and in IM (to 119 +/- 5.1 and 124.5 +/- 6.5% of controls, respectively). Western blot analysis revealed a cycloheximide-sensitive increase in StAR protein in mitochondrial extracts of Ca2+-clamped glomerulosa cells (to 159 +/- 23% of controls). In submitochondrial fractions, there was a selective accumulation of StAR protein in IM following stimulation with Ca2+ (228 +/- 50%). Similarly, Ang II increased StAR protein in IM, and this effect was prevented by cycloheximide. In contrast, neither Ca2+ nor Ang II had any effect on the submitochondrial distribution of cytochrome P450scc and 3beta-hydroxysteroid dehydrogenase isomerase. The intramitochondrial presence of the latter enzyme was further confirmed by immunogold staining in rat adrenal fasciculata cells and by immunoblot analysis in MA-10 mouse testicular Leydig cells. These findings demonstrate that under acute stimulation with Ca2+-mobilizing agents, newly synthesized StAR protein accumulates in IM after transiting through CS. Moreover, our results suggest that the import of StAR protein into IM may be associated with cholesterol transfer, thus promoting precursor supply to the two first enzymes of the steroidogenic cascade within the mitochondria and thereby activating mineralocorticoid synthesis.     

Calcium stimulates intramitochondrial cholesterol transfer in bovine adrenal glomerulosa cells

In adrenal glomerulosa cells, angiotensin II (Ang II) stimulates aldosterone synthesis through rises of cytosolic calcium ([Ca2+]c). The rate-limiting step in this process is the transfer of cholesterol to the inner mitochondrial membrane, where it is converted to pregnenolone by the P450 side chain cleavage enzyme. The aim of the present study was to examine the effect of changes in [Ca2+]c and of Ang II on intramitochondrial cholesterol distribution. Freshly prepared bovine zona glomerulosa cells were submitted to a cytosolic Ca2+ clamp (600 nM) or stimulated with Ang II (10 nM). Mitochondria were isolated and subfractionated into outer membranes (OM), inner membranes (IM), and contact sites (CS). Cholesterol content was determined by the cholesterol oxidase assay. Stimulation of intact cells with Ca2+ led to a marked decrease in cholesterol content of OM (to 54 +/- 24% of controls, n = 5) and to a concomitant increase of cholesterol in CS and IM (to 145 +/- 14%, n = 5). When glomerulosa cells were exposed to Ang II, a marked increase of cholesterol in CS occurred (to 172 +/- 16% of controls, n = 5). No significant changes were detected in OM cholesterol, suggesting a stimulation of cholesterol supply to the mitochondria in response to Ang II. Cycloheximide specifically and significantly reduced Ca2+-activated cholesterol transfer to CS and IM. In conclusion, our data indicate that one of the main functions of the Ca2+ messenger is to increase cholesterol supply to the P450 side chain cleavage enzyme by enhancing endogenous intermembrane cholesterol transfer to a mitochondrial site containing the enzymes responsible for the initial steps of the steroidogenic cascade.     

Parallel c-means algorithm for image segmentation on a reconfigurable mesh computer

In this paper, we propose a parallel algorithm for data classification, and its application for Magnetic Resonance Images (MRI) segmentation. The studied classification method is the well-known c-means method. The use of the parallel architecture in the classification domain is introduced in order to improve the complexities of the corresponding algorithms, so that they will be considered as a pre-processing procedure. The proposed algorithm is assigned to be implemented on a parallel machine, which is the reconfigurable mesh computer (RMC). The image of size (m × n) to be processed must be stored on the RMC of the same size, one pixel per processing element (PE).     

GPU fuzzy c-means algorithm implementations: performance analysis on medical image segmentation

Multimedia Tools and Applications - Image segmentation in the medical imagery such as MRI, is an essential step to the sensitive analysis of human tissues lesions with the objective to improve the...     

The performances of iterative type-2 fuzzy C-mean on GPU for image segmentation

The Journal of Supercomputing - Fuzzy C-mean (FCM) is an algorithm for data segmentation and classification, robust and very popular within the scientific community. It is used in several fields...     

Impact of IGBT behavior on design optimization of soft switchinginverter topologies

IGBTs operated under a zero voltage switching scheme exhibit significantly different operating characteristics from specified parameters in manufacturer data sheets. Important differences include an elevated tail current, dynamic saturation and resulting snubber dump, and a significantly higher conduction loss under moderate to high di/dt conditions. Soft switching inverter topologies of various kinds have been reported in the literature including resonant DC and AC link inverters, resonant pole inverters and many variations thereof. Loss characterization of soft switching inverter circuits reported to date is based on manufacturer data sheets, and do not reflect actual losses incurred in the circuit. Including the actual device behavior into the circuit loss calculations has a big impact on the total losses, and more importantly the trade-offs which impact the viability of a given topology at a specified operating point. This paper presents a detailed analysis of the losses in soft switching inverters and the impact on topology choice. A detailed design of a 50 kVA IGBT-based resonant DC link inverter is used as a design example, and it is seen that optimal design points and choices are significantly different from those that have been reported earlier