Glutathione deficiency down-regulates hepatic lipogenesis in rats

<p>Abstract</p> <p>Background</p> <p>Oxidative stress is supposed to increase lipid accumulation by stimulation of hepatic lipogenesis at transcriptional level. This study was performed to investigate the role of glutathione in the regulation of this process. For that purpose, male rats were treated with buthionine sulfoximine (BSO), a specific inhibitor of γ-glutamylcysteine synthetase, for 7 days and compared with untreated control rats.</p> <p>Results</p> <p>BSO treatment caused a significant reduction of total glutathione in liver (-70%), which was attributable to diminished levels of reduced glutathione (GSH, -71%). Glutathione-deficient rats had lower triglyceride concentrations in their livers than the control rats (-23%), whereas the circulating triglycerides and the cholesterol concentrations in plasma and liver were not different between the two groups of rats. Livers of glutathione-deficient rats had lower mRNA abundance of sterol regulatory element-binding protein (SREBP)-1c (-47%), Spot (S)14 (-29%) and diacylglycerol acyltransferase 2 (DGAT-2, -27%) and a lower enzyme activity of fatty acid synthase (FAS, -26%) than livers of the control rats. Glutathione-deficient rats had also a lower hepatic activity of the redox-sensitive protein-tyrosine phosphatase (PTP)1B, and a higher concentration of irreversible oxidized PTP1B than control rats. No differences were observed in protein expression of total PTP1B and the mature mRNA encoding active XBP1s, a key regulator of unfolded protein and ER stress response.</p> <p>Conclusion</p> <p>This study shows that glutathione deficiency lowers hepatic triglyceride concentrations via influencing lipogenesis. The reduced activity of PTP1B and the higher concentration of irreversible oxidized PTP1B could be, at least in part, responsible for this effect.</p

hsa‐miR‐374b‐5p regulates expression of the gene U2AF homology motif (UHM) kinase 1

Objective: We aimed to identify a microRNA (miRNA) that is significantly upregulated in blood and in cells of the oral mucosa upon exposure to the periodontitis main risk factors oral inflammation and tobacco smoke, to subsequently identify its target gene and to describe the molecular mechanism of gene regulation. Background: miRNAs are associated with many disorders. Array-based miRNA expression studies indicated a number of differentially expressed miRNAs in the pathology of oral diseases. However, these miRNAs mostly lacked replication, and their target genes have remained unknown. Methods: 863 miRNAs were analyzed in blood from 18 PD cases and 70 controls (Geniom Biochip). Selected miRNAs were analyzed for upregulation in the inflamed oral mucosa of PD patients using published miRNA expression profiling studies from gingival cells. hsa-miR-374b-5p mimic was overexpressed in primary gingival fibroblasts (pGFs) from 3 donors, and genome-wide mRNA expression was quantified (Clarion Array). Gene-specific regulation was validated by qRT-PCR and Luciferase activity in HeLa cells. Results: hsa-miR-374b-5p showed >twofold change (FC) in 3 independent studies performed in blood, gingival tissues, and cells. After hsa-miR-374b-5p overexpression, genome-wide expression analysis showed UHMK1 as top 1 downregulated gene in pGFs (p = 2.5 × 10-04 , fold change = -1.8). Reporter genes demonstrated that hsa-miR-374b-5p downregulates mRNA levels (p = .02; FC = -1.5), leading to reduction in protein activity (p = .013, FC = -1.3). Conclusions: hsa-miR-374b-5p is upregulated in blood and ginvial cells exposed to oral inflammation and tobacco smoke and regulates UHMK1, which has a role in osteoclast differentiation

A Vaccinia-based system for directed evolution of GPCRs in mammalian cells

Directed evolution in bacterial or yeast display systems has been successfully used to improve stability and expression of G protein-coupled receptors for structural and biophysical studies. Yet, several receptors cannot be tackled in microbial systems due to their complex molecular composition or unfavorable ligand properties. Here, we report an approach to evolve G protein-coupled receptors in mammalian cells. To achieve clonality and uniform expression, we develop a viral transduction system based on Vaccinia virus. By rational design of synthetic DNA libraries, we first evolve neurotensin receptor 1 for high stability and expression. Second, we demonstrate that receptors with complex molecular architectures and large ligands, such as the parathyroid hormone 1 receptor, can be readily evolved. Importantly, functional receptor properties can now be evolved in the presence of the mammalian signaling environment, resulting in receptor variants exhibiting increased allosteric coupling between the ligand binding site and the G protein interface. Our approach thus provides insights into the intricate molecular interplay required for GPCR activation

Nonperturbative Effects in Gluon Radiation and Photoproduction of Quark Pairs

We introduce a nonperturbative interaction for light-cone fluctuations containing quarks and gluons. The $\bar qq$ interaction squeezes the transverse size of these fluctuations in the photon and one does not need to simulate this effect via effective quark masses. The strength of this interaction is fixed by data. Data on diffractive dissociation of hadrons and photons show that the nonperturbative interaction of gluons is much stronger. We fix the parameters for the nonperturbative quark-gluon interaction by data for diffractive dissociation to large masses (triple-Pomeron regime). This allows us to predict nuclear shadowing for gluons which turns out to be not as strong as perturbative QCD predicts. We expect a delayed onset of gluon shadowing at $x \leq 10^{-2}$ shadowing of quarks. Gluon shadowing turns out to be nearly scale invariant up to virtualities $Q^2\sim 4 GeV^2$ due to presence of a semihard scale characterizing the strong nonperturbative interaction of gluons. We use the same concept to improve our description of gluon bremsstrahlung which is related to the distribution function for a quark-gluon fluctuation and the interaction cross section of a $\bar qqG$ fluctuation with a nucleon. We expect the nonperturbative interaction to suppress dramatically the gluon radiation at small transverse momenta compared to perturbative calculations.Comment: 58 pages of Latex including 11 figures. Shadowing for soft gluons and Fig. 6 are added as well as a few reference

On the Energy Dependence of the Dipole-Proton Cross Section in Deep Inelastic Scattering

We study the dipole picture of high-energy virtual-photon-proton scattering. It is shown that different choices for the energy variable in the dipole cross section used in the literature are not related to each other by simple arguments equating the typical dipole size and the inverse photon virtuality, contrary to what is often stated. We argue that the good quality of fits to structure functions that use Bjorken-x as the energy variable - which is strictly speaking not justified in the dipole picture - can instead be understood as a consequence of the sign of scaling violations that occur for increasing Q^2 at fixed small x. We show that the dipole formula for massless quarks has the structure of a convolution. From this we obtain derivative relations between the structure function F_2 at large and small Q^2 and the dipole-proton cross section at small and large dipole size r, respectively.Comment: 27 page

Characterization of the radiation tolerance of cryogenic diodes for the High Luminosity LHC inner triplet circuit

Cryogenic bypass diodes are part of the baseline powering layout for the circuits of the new Nb3Sn based final focus magnets of the high luminosity Large Hadron Collider. They will protect the magnets against excessive transient voltages during a nonuniform quenching process. The diodes are located inside an extension to the magnet cryostat, operated in superfluid helium and exposed to ionizing radiation. Therefore, the radiation tolerance of different types of diodes has been tested at cryogenic temperatures in CERN’s CHARM irradiation test facility during its 2018 run. The forward bias characteristics, the turn-on voltage and the reverse blocking voltage of each diode were measured weekly at 4.2 K and 77 K, as a function of the accumulated radiation dose. The diodes were submitted to a total dose close to 12 kGy and a 1 MeV neutron equivalent fluence of 2.2×1014  cm−2. After the end of the irradiation program the annealing behavior of the diodes was tested by increasing the temperature slowly to 293 K. This paper describes the experimental setup, the measurement procedure and the analysis of the measurements performed during the irradiation program as well as the results of the annealing study

Evidence from Studies with Heat-Stressed Caco-2 Cells, C. elegans and Growing Broilers

Climatic changes and heat stress have become a great challenge in the livestock industry, negatively affecting, in particular, poultry feed intake and intestinal barrier malfunction. Recently, phytogenic feed additives were applied to reduce heat stress effects on animal farming. Here, we investigated the effects of ginseng extract using various in vitro and in vivo experiments. Quantitative real-time PCR, transepithelial electrical resistance measurements and survival assays under heat stress conditions were carried out in various model systems, including Caco-2 cells, Caenorhabditis elegans and jejunum samples of broilers. Under heat stress conditions, ginseng treatment lowered the expression of HSPA1A (Caco-2) and the heat shock protein genes hsp-1 and hsp-16.2 (both in C. elegans), while all three of the tested genes encoding tight junction proteins, CLDN3, OCLN and CLDN1 (Caco-2), were upregulated. In addition, we observed prolonged survival under heat stress in Caenorhabditis elegans, and a better performance of growing ginseng-fed broilers by the increased gene expression of selected heat shock and tight junction proteins. The presence of ginseng extract resulted in a reduced decrease in transepithelial resistance under heat shock conditions. Finally, LC-MS analysis was performed to quantitate the most prominent ginsenosides in the extract used for this study, being Re, Rg1, Rc, Rb2 and Rd. In conclusion, ginseng extract was found to be a suitable feed additive in animal nutrition to reduce the negative physiological effects caused by heat stress. View Full-Tex

SKP2 attenuates autophagy through Beclin1-ubiquitination and its inhibition reduces MERS-Coronavirus infection

Autophagy is an essential cellular process affecting virus infections and other diseases and Beclin1 (BECN1) is one of its key regulators. Here, we identified S-phase kinase-associated protein 2 (SKP2) as E3 ligase that executes lysine-48-linked poly-ubiquitination of BECN1, thus promoting its proteasomal degradation. SKP2 activity is regulated by phosphorylation in a hetero-complex involving FKBP51, PHLPP, AKT1, and BECN1. Genetic or pharmacological inhibition of SKP2 decreases BECN1 ubiquitination, decreases BECN1 degradation and enhances autophagic flux. Middle East respiratory syndrome coronavirus (MERS-CoV) multiplication results in reduced BECN1 levels and blocks the fusion of autophagosomes and lysosomes. Inhibitors of SKP2 not only enhance autophagy but also reduce the replication of MERS-CoV up to 28,000-fold. The SKP2-BECN1 link constitutes a promising target for host-directed antiviral drugs and possibly other autophagy-sensitive conditions