Cytochrome P450 endoplasmic reticulum-associated degradation (ERAD): therapeutic and pathophysiological implications.

The hepatic endoplasmic reticulum (ER)-anchored cytochromes P450 (P450s) are mixed-function oxidases engaged in the biotransformation of physiologically relevant endobiotics as well as of myriad xenobiotics of therapeutic and environmental relevance. P450 ER-content and hence function is regulated by their coordinated hemoprotein syntheses and proteolytic turnover. Such P450 proteolytic turnover occurs through a process known as ER-associated degradation (ERAD) that involves ubiquitin-dependent proteasomal degradation (UPD) and/or autophagic-lysosomal degradation (ALD). Herein, on the basis of available literature reports and our own recent findings of in vitro as well as in vivo experimental studies, we discuss the therapeutic and pathophysiological implications of altered P450 ERAD and its plausible clinical relevance. We specifically (i) describe the P450 ERAD-machinery and how it may be repurposed for the generation of antigenic P450 peptides involved in P450 autoantibody pathogenesis in drug-induced acute hypersensitivity reactions and liver injury, or viral hepatitis; (ii) discuss the relevance of accelerated or disrupted P450-ERAD to the pharmacological and/or toxicological effects of clinically relevant P450 drug substrates; and (iii) detail the pathophysiological consequences of disrupted P450 ERAD, contributing to non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH) under certain synergistic cellular conditions

Growth of superconducting MgB2 thin films via postannealing techniques

We report the effect of annealing on the superconductivity of MgB2 thin films as functions of the postannealing temperature in the range from 700 C to 950 C and of the postannealing time in the range from 30 min to 120 min. On annealing at 900 C for 30 min, we obtained the best-quality MgB2 films with a transition temperature of 39 K and a critical current density of ~ 10^7 A/cm^2. Using the scanning electron microscopy, we also investigated the film growth mechanism. The samples annealed at higher temperatures showed the larger grain sizes, well-aligned crystal structures with preferential orientations along the c-axis, and smooth surface morphologies. However, a longer annealing time prevented the alignment of grains and reduced the superconductivity, indicating a strong interfacial reaction between the substrate and the MgB2 film.Comment: 7 pages, 4 figures include

Hepatic cytochromes P450: structural degrons and barcodes, posttranslational modifications and cellular adapters in the ERAD-endgame.

The endoplasmic reticulum (ER)-anchored hepatic cytochromes P450 (P450s) are enzymes that metabolize endo- and xenobiotics i.e. drugs, carcinogens, toxins, natural and chemical products. These agents modulate liver P450 content through increased synthesis or reduction via inactivation and/or proteolytic degradation, resulting in clinically significant drug-drug interactions. P450 proteolytic degradation occurs via ER-associated degradation (ERAD) involving either of two distinct routes: Ubiquitin (Ub)-dependent 26S proteasomal degradation (ERAD/UPD) or autophagic lysosomal degradation (ERAD/ALD). CYP3A4, the major human liver/intestinal P450, and the fast-turnover CYP2E1 species are degraded via ERAD/UPD entailing multisite protein phosphorylation and subsequent ubiquitination by gp78 and CHIP E3 Ub-ligases. We are gaining insight into the nature of the structural determinants involved in CYP3A4 and CYP2E1 molecular recognition in ERAD/UPD [i.e. K48-linked polyUb chains and linear and/or "conformational" phosphodegrons consisting either of consecutive sequences on surface loops and/or disordered regions, or structurally-assembled surface clusters of negatively charged acidic (Asp/Glu) and phosphorylated (Ser/Thr) residues, within or vicinal to which, Lys-residues are targeted for ubiquitination]. Structural inspection of select human liver P450s reveals that such linear or conformational phosphodegrons may indeed be a common P450-ERAD/UPD feature. By contrast, although many P450s such as the slow-turnover CYP2E1 species and rat liver CYP2B1 and CYP2C11 are degraded via ERAD/ALD, little is known about the mechanism of their ALD-targeting. On the basis of our current knowledge of ALD-substrate targeting, we propose a tripartite conjunction of K63-linked Ub-chains, P450 structural "LIR" motifs and selective cellular "cargo receptors" as plausible P450-ALD determinants

Integrated Information System for Sustainable Urban Regeneration

Information systems are widely used in urban planning process for communication between different side actors. However, most of them have been implemented without providing possibilities for decision makers to participate together with urban planners in the process. This research aims to outline a framework where an interactive model for decision making plays a key role in creating a collaborative environment. The proposal regards the information representation as the main instrument for encouraging a constructive dialog between different actors. The focus is on the relationship between three elements of information representation: level of detail, type of visualization and interaction. Combining these elements, information can be provided in a dynamic way enabling more effective exploration and understanding. The proposed strategy implements a digital model that operates on different scales and levels in order to support the key stages of the planning process for sustainable urban regeneration in Bulgaria. Positional approach is used to define the functionality and decision making operation for the selected process. As a result research ideas about the use of the digital model are presented

Utilization of Laboratory Tests for Tuberculosis and Mycobacterial Disease in Korea

AbstractObjectivesIn Korea, a large portion of tuberculosis (TB) patients are diagnosed and treated in private institutes. Laboratory tests are crucial for TB control. There are many possible problems using laboratory tests in the private sector. In this study, we aimed to investigate the characteristics and trends of utilizing laboratory tests for TB and mycobacterial diseases in the private sector by analyzing the National Health Insurance (NHI) database.MethodsAfter selecting TB or other mycobacteria-related test items, we searched the number and cost of each item on the website of the Health Insurance Review and Assessment Service using the code of each test from 2007 to 2012.ResultsOur data revealed that the number and cost of tests drastically increased between 2007 and 2012. Culture and molecular tests primarily contributed to the tremendous increases. For each year, concentrated smearing and fluorochrome staining were more commonly used. The number of serologic tests for latent TB infection stagnated, despite the expansion of contact investigation.ConclusionThe NHI data could be considerably useful for understanding the utilization trends of laboratory tests for TB and mycobacterial diseases in Korea. Our data showed that TB laboratory systems have recently improved. In this study, many issues were noticed. Therefore, solutions to these issues are required and the continued monitoring of NHI data regarding laboratory diagnosis

Simultaneous deletion of floxed genes mediated by CaMKIIa-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Go-alfa

The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice generated by crossing CaMKIIα-Cre+/Cre mice with floxed ROSA26 lacZ reporter (Rosa26+/stop-lacZ) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26+/stop-lacZ::CaMKIIα-Cre+/Cre mice. Similarly, when double transgenic Gnao+/f::CaMKIIα-Cre+/Cre mice carrying a floxed Go-alpha gene (Gnaof/f) were backcrossed to wild-type females, approximately 22% of the offspring carried the disrupted allele (GnaoΔ) without inheriting the Cre transgene. The GnaoΔ/Δ mice closely resembled conventional Go-alpha knockout mice (Gnao−/−) with respect to impairment of their behavior. Thus, we conclude that CaMKIIα-Cre mice afford recombination for both tissue- and time-controlled inactivation of floxed target genes in the brain and for their permanent disruption. This work also emphasizes that extra caution should be exercised in utilizing CaMKIIα-Cre mice as breeding pairs.Fil: Choi, Chan-Il. Ajou University. School of Medicine; Corea del SurFil: Yoon, Sang-Phil. Ajou University. School of Medicine; Corea del SurFil: Choi, Jung-Mi. Ajou University. School of Medicine; Corea del SurFil: Kim, Sung-Soo. Ajou University. School of Medicine; Corea del SurFil: Lee, Young-Don. Ajou University. School of Medicine; Corea del SurFil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences; Estados Unidos. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Suh-Kim. Haeyoung. Ajou University. School of Medicine; Corea del Su