Generation and characterization of a mitotane-resistant adrenocortical cell line

Mitotane is the only drug approved for the therapy of adrenocortical carcinoma (ACC). Its clinical use is limited by the occurrence of relapse during therapy. To investigate the underlying mechanisms in vitro, we here generated mitotane-resistant cell lines. After long-term pulsed treatment of HAC-15 human adrenocortical carcinoma cells with 70 µM mitotane, we isolated monoclonal cell populations of treated cells and controls and assessed their respective mitotane sensitivities by MTT assay. We performed exome sequencing and electron microscopy, conducted gene expression microarray analysis and determined intracellular lipid concentrations in the presence and absence of mitotane. Clonal cell lines established after pulsed treatment were resistant to mitotane (IC50 of 102.2 ± 7.3 µM (n = 12) vs 39.4 ± 6.2 µM (n = 6) in controls (biological replicates, mean ± s.d., P = 0.0001)). Unlike nonresistant clones, resistant clones maintained normal mitochondrial and nucleolar morphology during mitotane treatment. Resistant clones largely shared structural and single nucleotide variants, suggesting a common cell of origin. Resistance depended, in part, on extracellular lipoproteins and was associated with alterations in intracellular lipid homeostasis, including levels of free cholesterol, as well as decreased steroid production. By gene expression analysis, resistant cells showed profound alterations in pathways including steroid metabolism and transport, apoptosis, cell growth and Wnt signaling. These studies establish an in vitro model of mitotane resistance in ACC and point to underlying molecular mechanisms. They may enable future studies to overcome resistance in vitro and improve ACC treatment in vivo

Evaluation of a Sodium–Water Reaction Event Caused by Steam Generator Tubes Break in the Prototype Generation IV Sodium-cooled Fast Reactor

AbstractThe prototype generation IV sodium-cooled fast reactor (PGSFR) has been developed by the Korea Atomic Energy Research Institute. This reactor uses sodium as a reactor coolant to transfer the core heat energy to the turbine. Sodium has chemical characteristics that allow it to violently react with materials such as a water or steam. When a sodium–water reaction (SWR) occurs due to leakage or breakage of steam generator tubes, high-pressure waves and corrosive reaction products are produced, which threaten the structural integrity of the components of the intermediate heat-transfer system (IHTS) and the safety of the primary heat-transfer system (PHTS). In the PGSFR, SWR events are included in the design-basis event. This event should be analyzed from the viewpoint of the integrities of the IHTS and fuel rods. To evaluate the integrity of the IHTS based on the consequences of the SWR, the behaviors of the generated high-pressure waves are analyzed at the major positions of a failed IHTS loop using a sodium–water advanced analysis method-II code. The integrity of the fuel rods must be consistently maintained below the safety acceptance criteria to avoid the consequences of the SWR. The integrity of the PHTS is evaluated using the multidimensional analysis of reactor safety-liquid metal reactor code to model the whole plant

Temperature Dependence of Conductance and Plateau Length for Single-Molecule Junctions Formed with Silver Electrodes

The temperature dependence of the conductance and plateau length for the single-molecule junctions formed with Ag electrodes was investigated by the scanning tunneling microscope-based break-junction technique. The electrode–electrode gap distance (GD) was measured by varying temperature as soon as the Ag metal contact was ruptured. The compressed molecular junction (a tilted molecule) with a small GD at low temperature showed a higher conductance and longer plateau than the vertically stretched molecular junction with a large GD at high temperature. However, for the Au electrodes, the GD did not change by varying temperature, resulting in a constant conductance and plateau. These results are attributed to a larger diffusion constant of Ag compared to Au, yielding a relatively temperature-dependent GD for the Ag electrode. This study may advance the understanding of the electrical and mechanical properties in single-molecule-based devices by varying temperature

A Preliminary Safety Analysis for the Prototype Gen IV Sodium-Cooled Fast Reactor

Korea Atomic Energy Research Institute has been developing a pool-type sodium-cooled fast reactor of the Prototype Gen-IV Sodium-cooled Fast Reactor (PGSFR). To assess the effectiveness of the inherent safety features of the PGSFR, the system transients during design basis accidents and design extended conditions are analyzed with MARS-LMR and the subchannel blockage events are analyzed with MATRA-LMR-FB. In addition, the in-vessel source term is calculated based on the super-safe, small, and simple reactor methodology. The results show that the PGSFR meets safety acceptance criteria with a sufficient margin during the events and keeps accidents from deteriorating into more severe accidents

Contact Effect of ReS₂/Metal Interface

Rhenium disulfide (ReS₂) has attracted immense interest as a promising two-dimensional material for optoelectronic devices owing to its outstanding photonic response based on its energy band gap’s insensitivity to the layer thickness. Here, we theoretically calculated the electrical band structure of mono-, bi-, and trilayer ReS₂ and experimentally found the work function to be 4.8 eV, which was shown to be independent of the layer thickness. We also evaluated the contact resistance of a ReS₂ field-effect transistor using a <i>Y</i>-function method with various metal electrodes, including graphene. The ReS₂ channel is a strong n-type semiconductor, thus a lower work function than that of metals tends to lead to a lower contact resistance. Moreover, the graphene electrodes, which were not chemically or physically bonded to ReS₂, showed the lowest contact resistance, regardless of the work function, suggesting a significant Fermi-level pinning effect at the ReS₂/metal interface. In addition, an asymmetric Schottky diode device was demonstrated using Ti or graphene for ohmic contacts and Pt or Pd for Schottky contacts. The ReS₂-based transistor used in this study on the work function of ReS₂ achieved the possibility of designing the next-generation nanologic devices

Disease-specific eQTL screening reveals an anti-fibrotic effect of AGXT2 in non-alcoholic fatty liver disease

Background &amp; Aims: Non-alcoholic fatty liver disease (NAFLD) poses an increasing clinical burden. Genome-wide association studies have revealed a limited contribution of genomic variants to the disease, requiring alternative but robust approaches to identify disease-associated variants and genes. We carried out a disease-specific expression quantitative trait loci (eQTL) screen to identify novel genetic factors that specifically act on NAFLD progression on the basis of genotype. Methods: We recruited 125 Korean patients (83 with biopsy proven NAFLD and 42 without NAFLD) and performed eQTL analyses using 21,272 transcripts and 3,234,941 genotyped and imputed single nucleotide polymorphisms. We then selected eQTLs that were detected only in the NAFLD group, but not in the control group (i.e., NAFLD-eQTLs). An additional cohort of 162 Korean individuals with NAFLD was used for replication. The function of the selected eQTL toward NAFLD development was validated using HepG2, primary hepatocytes and NAFLD mouse models. Results: The NAFLD-specific eQTL screening yielded 242 loci. Among them, AGXT2, encoding alanine-glyoxylate aminotransferase 2, displayed decreased expression in patients with NAFLD homozygous for the non-reference allele of rs2291702, compared to no-NAFLD individuals with the same genotype (p = 4.79 x 10(-6)). This change was replicated in an additional 162 individuals, yielding a combined p value of 8.05 x 10(-8) from a total of 245 patients with NAFLD and 42 controls. Knockdown of AGXT2 induced palmitate-overloaded hepatocyte death by increasing endoplasmic reticulum stress, and exacerbated NAFLD diet-induced liver fibrosis in mice, while overexpression of AGXT2 attenuated liver fibrosis and steatosis. Conclusions: We identified a new molecular role for AGXT2 in NAFLD. Our overall approach will serve as an efficient tool for uncovering novel genetic factors that contribute to liver steatosis and fibrosis in patients with NAFLD. Lay summary: Elucidating causal genes for non-alcoholic fatty liver disease (NAFLD) has been challenging due to limited tissue availability and the polygenic nature of the disease. Using liver and blood samples from 125 Korean individuals (83 with NAFLD and 42 without NAFLD), we devised a new analytic method to identify causal genes. Among the candidates, we found that AGXT2-rs2291702 protects against liver fibrosis in a genotype dependent manner with the potential for therapeutic interventions. Our approach enables the discovery of causal genes that act on the basis of genotype. (C) 2021 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.N

CLCN2 chloride channel mutations in familial hyperaldosteronism type II

Primary aldosteronism, a common cause of severe hypertension1, features constitutive production of the adrenal steroid aldosterone. We analyzed a multiplex family with familial hyperaldosteronism type II (FH-II)2 and 80 additional probands with unsolved early-onset primary aldosteronism. Eight probands had novel heterozygous variants in CLCN2, including two de novo mutations and four independent occurrences of a mutation encoding an identical p.Arg172Gln substitution; all relatives with early-onset primary aldosteronism carried the CLCN2 variant found in the proband. CLCN2 encodes a voltage-gated chloride channel expressed in adrenal glomerulosa that opens at hyperpolarized membrane potentials. Channel opening depolarizes glomerulosa cells and induces expression of aldosterone synthase, the rate-limiting enzyme for aldosterone biosynthesis. Mutant channels show gain of function, with higher open probabilities at the glomerulosa resting potential. These findings for the first time demonstrate a role of anion channels in glomerulosa membrane potential determination, aldosterone production and hypertension. They establish the cause of a substantial fraction of early-onset primary aldosteronism