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.     

The employment of strontium to activate mouse oocytes: effects on spermatid-injection outcome

The present research investigated the effects of various strontium concentrations, in combination with different incubation periods, on mouse parthenogentic oocyte activation and blastocyst development. The results for blastocyst development showed a trend indicating that 10 mM strontium for 3 h was the optimal strontium protocol. Ethanol, an agent that incites oocyte activation via a monotonic rise in calcium, was employed as a control. The outcome of blastocyst formation arising from parthenogenic ethanol activation was significantly less (P < 0.001) than that achieved by the optimal strontium protocol. To assess the impact of strontium oocyte activation on embryo viability following fertilization with immature germ cells, the protocol of 10 mM strontium for 3 h was applied to oocytes injected with round spermatids and then compared with other protocols. The results indicate that following round-spermatid injection the benefits derived from strontium artificial oocyte activation are evident during both pre- and post-implantation development. However, in order to adjust the protocol to the most effective round-spermatid injection in relation to the oocyte cell cycle, injection was done 1.5 h after strontium activation followed by another 1.5 h activation in strontium. The implementation of round-spermatid injection in combination with this oocyte-activation protocol led to live-birth outcomes not significantly different to those outcomes obtained by mature spermatozoa.     

Global Shutter CMOS Image Sensor With Wide Dynamic Range

A novel concept for global shutter CMOS image sensors with wide dynamic range (WDR) implementation is presented. The proposed imager is based on the multisampling WDR approach and it allows an efficient global shutter pixel implementation achieving small pixel size and high fill factor. The proposed imager provides wide DR by applying adaptive exposure time to each pixel, according to the local illumination intensity level. Two pixel configurations, employing different kinds of a 1-bit in-pixel memory were implemented. An imager, including two different pixels was designed and simulated in 0.18-mum CMOS technology. System architecture and operation are discussed and simulation results are presented.     

An APS with 2-D winner-take-all selection employing adaptive spatial filtering and false alarm reduction

An active pixel sensor (APS) with two-dimensional winner-take-all (WTA) detection is presented. This system-on-a-chip employs adaptive spatial filtering of the processed image, with bad pixel elimination and false alarm reduction in case of a missing object. The circuit has a unique ability of adaptive spatial filtering that allows removal of the background from the image, one stage before it is transferred to the WTA detection circuit. A test chip of a 64/sup */64 array has been implemented in 0.5-/spl mu/m CMOS technology, has a 49% fill factor, is operated by a 3.3-V supply, and dissipates 36 mW at video rate. System architecture and operation are discussed and measurements from the prototype chip are presented.     

Ethane dimethane sulfonate and NNN'N'-tetrakis-(2-pyridylmethyl)ethylenediamine inhibit steroidogenic acute regulatory (StAR) protein expression in MA-10 Leydig cells and rat Sertoli cells

Apoptosis inhibits steroid biosynthesis, but it is not clear how the Steroidogenic Acute Regulatory (StAR) protein, is affected. To characterize StAR expression during apoptosis, mouse MA-10 Leydig tumor cells were treated with ethane dimethane sulfonate (EDS), an inducer of apoptosis, and the metal ion chelator NNN'N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN), an inducer of cell death. Both chemicals induced cell death and similarly inhibited dbcAMP-stimulated steroidogenesis and accumulation of the 30 kDa form of StAR. Utilizing the dye JC-1, it was found that TPEN and EDS also impaired the mitochondrial electrochemical potential (delta psi). In Sertoli cells, which also express StAR, EDS induced cell death and attenuated StAR expression. We conclude 1) steroidogenesis and accumulation of mature StAR protein are inhibited as a consequence of the induction of apoptosis; 2) reduced levels of StAR may be partially attributed to inhibition of import because of the loss of delta psi; 3) loss of steroidogenesis is probably due to loss of StAR synthesis and disruption of delta psi.     

Coralline Skeleton Biomaterial Reduces Phagocytosis in Mouse Blood in vitro

Inflammatory and immunogenic response to foreign bodies presents a challenge in the use of biomaterials as implants for tissue restoration. Therefore, there is a need to understand the interactions between such implants and the blood. One such material, currently in clinical use for bone replacement in humans, is the skeleton of corals, in the form of crystalline aragonite. This biomaterial has been shown to impart a protective and supportive influence on several types of cells ex vivo. The carbonate skeleton activates secretion in phagocytes in vitro, however its effects on these cells in the blood, and on the process of phagocytosis itself, remain unknown. Using 1–500 μm particles of coral skeleton, we show that these particles bind blood proteins and alter the leukocyte population, reducing the proportion of granulocytes by more than 3-fold with no effect on the proportion of monocytes. In addition, the presence of coral skeleton in the blood causes a reduction in phagocytosis. Specifically, we observed a decrease in the percentage of phagocytic cells by 27 % in the granulocytes and by 73 % in monocyte family, as well as a 41.6 % reduction in the MFI of granulocytes, but with no such effect on monocytes. Taken together, the results suggest that the coral skeleton biomaterial may act as a strong, promotive scaffold for tissue regeneration due to its ability to reduce its rejection by inflammatory reactions such as phagocytosis.     

Transformational leadership and group interaction as climate antecedents: A social network analysis.

In order to test the social mechanisms through which organizational climate emerges, this article introduces a model that combines transformational leadership and social interaction as antecedents of climate strength (i.e., the degree of within-unit agreement about climate perceptions). Despite their longstanding status as primary variables, both antecedents have received limited empirical research. The sample consisted of 45 platoons of infantry soldiers from 5 different brigades, using safety climate as the exemplar. Results indicate a partially mediated model between transformational leadership and climate strength, with density of group communication network as the mediating variable. In addition, the results showed independent effects for group centralization of the communication and friendship networks, which exerted incremental effects on climate strength over transformational leadership. Whereas centralization of the communication network was found to be negatively related to climate strength, centralization of the friendship network was positively related to it. Theoretical and practical implications are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Borders: An Efficient Algorithm for Association Generation in Dynamic Databases

We consider the problem of finding association rules in a database with binary attributes. Most algorithms for finding such rules assume that all the data is available at the start of the data mining session. In practice, the data in the database may change over time, with records being added and deleted. At any given time, the rules for the current set of data are of interest. The naive, and highly inefficient, solution would be to rerun the association generation algorithm from scratch following the arrival of each new batch of data. This paper describes the Borders algorithm, which provides an efficient method for generating associations incrementally, from dynamically changing databases. Experimental results show an improved performance of the new algorithm when compared with previous solutions to the problem.