Detection of hCG Responsive Expression of the Steroidogenic Acute Regulatory Protein in Mouse Leydig Cells

The steroidogenic acute regulatory (StAR) protein, a novel mitochondrial protein, is involved in the regulation of steroid hormone biosynthesis through its mediation of the intramitochondrial transport of the steroid substrate, cholesterol, to the cytochrome P450 cholesterol side chain cleavage (P450scc) enzyme. The expression of StAR protein is regulated by cAMP-dependent signaling in steroidogenic cells. During the course of our studies in mouse Leydig cells, we employ several methods for studying the regulation of StAR protein expression by human chorionic gonadotropin (hCG). A sensitive quantitative reverse transcription and polymerase chain reaction (RT-PCR) was utilized for determining StAR mRNA expression. Stimulation of mLTC-1 mouse Leydig tumor cells with hCG resulted in the coordinate regulation of StAR mRNA expression and progesterone accumulation in a time-response manner. The validity and accuracy of quantitative RT-PCR results in mLTC-1 cells were verified by a competitive PCR approach and were further confirmed in primary cultures of isolated mouse Leydig cells. Immunoblotting studies demonstrated an increase in the levels of the StAR protein in a concentration dependent manner following hCG stimulation in mLTC-1 cells. Northern hybridization analysis revealed three StAR transcripts, all of which were of sufficient size to encode functional StAR protein, and which were coordinately expressed in response to hCG. Collectively, the experimental approaches utilized in the present investigation allow for the demonstration and characterization of hCG mediated regulation of StAR mRNA and StAR protein expression in mouse Leydig cells

Advanced production routes for metal matrix composites

The use of metal matrix composites (MMCs) in a variety of products is significantly increasing with time due to the fact that their properties can be tailored and designed to suit specific applications. However, the future usage of MMC products is very much dependent on their beneficial aspects and hence it is critical to ensure in a robust repeatable manner the superior physical property advantages compared to conventional unreinforced monolithic metal counterparts. Although numerous routes are available for production of MMC products, each of them has their own advantages and disadvantages. This article provides an overview of advanced production routes for MMCs. The discussion also highlights challenges and presents a future prospectus for MMCs. Powder metallurgy and casting routes are still extensively used for production of MMCs. Aluminum alloys are today the most commonly used matrix materials in MMC products. Carbides (eg, SiC, TiC, and B4C), carbon allotropes (eg, CNTs and graphene), and alumina (Al2O3) are currently the most used reinforcement materials. Nevertheless, the use of nano and of hybrid reinforcements are seeing increased usage in niche applications. Additive manufacturing (AM) is discussed as a novel production route for MMC products. This process represents a promising method for the production of MMC products