Helix 11 Dynamics Is Critical for Constitutive Androstane Receptor Activity

SummaryThe constitutive androstane receptor (CAR) transactivation can occur in the absence of exogenous ligand and this activity is enhanced by agonists TCPOBOP and meclizine. We use biophysical and cell-based assays to show that increased activity of CAR(TCPOBOP) relative to CAR(meclizine) corresponds to a higher affinity of CAR(TCPOBOP) for the steroid receptor coactivator-1. Additionally, steady-state fluorescence spectra suggest conformational differences between CAR(TCPOBOP):RXR and CAR(meclizine):RXR. Hydrogen/deuterium exchange (HDX) data indicate that the CAR activation function 2 (AF-2) is more stable in CAR(TCPOBOP):RXR and CAR(meclizine):RXR than in CAR:RXR. HDX kinetics also show significant differences between CAR(TCPOBOP):RXR and CAR(meclizine):RXR. Unlike CAR(meclizine):RXR, CAR(TCPOBOP):RXR shows a higher overall stabilization that extends into RXR. We identify residues 339–345 in CAR as an allosteric regulatory site with a greater magnitude reduction in exchange kinetics in CAR(TCPOBOP):RXR than CAR(meclizine):RXR. Accordingly, assays with mutations on CAR at leucine-340 and leucine-343 confirm this region as an important determinant of CAR activity

Lrg1 Regulates β (1, 3)-Glucan Masking in Candida albicans through the Cek1 MAP Kinase Pathway

Candida albicans is among the most prevalent opportunistic human fungal pathogens. The ability to mask the immunogenic polysaccharide β (1,3)-glucan from immune detection via a layer of mannosylated proteins is a key virulence factor of C. albicans. We previously reported that hyperactivation of the Cek1 mitogen-activated protein (MAP) kinase pathway promotes β (1,3)-glucan exposure. In this communication, we report a novel upstream regulator of Cek1 activation and characterize the impact of Cek1 activity on fungal virulence. Lrg1 encodes a GTPase-activating protein (GAP) that has been suggested to inhibit the GTPase Rho1. We found that disruption of LRG1 causes Cek1 hyperactivation and β (1,3)-glucan unmasking. However, when GTPase activation was measured for a panel of GTPases, the lrg1ΔΔ mutant exhibited increased activation of Cdc42 and Ras1 but not Rho1 or Rac1. Unmasking and Cek1 activation in the lrg1ΔΔ mutant can be blocked by inhibition of the Ste11 MAP kinase kinase kinase (MAPKKK), indicating that the lrg1ΔΔ mutant acts through the canonical Cek1 MAP kinase cascade. In order to determine how Cek1 hyperactivation specifically impacts virulence, a doxycycline-repressible hyperactive STE11ΔN467 allele was expressed in C. albicans. In the absence of doxycycline, this allele overexpressed STE11ΔN467, which induced production of proinflammatory tumor necrosis factor alpha (TNF-α) from murine macrophages. This in vitrophenotype correlates with decreased colonization and virulence in a mouse model of systemic infection. The mechanism by which Ste11ΔN467 causes unmasking was explored with RNA sequencing (RNA-Seq) analysis. Overexpression of Ste11ΔN467 caused upregulation of the Cph1 transcription factor and of a group of cell wall-modifying proteins which are predicted to impact cell wall architecture

Biomonitoring of metal deposition in northern Spain by moss analysis

The results of the first survey carried out in northern Spain to determine atmospheric deposition of metals by analysis of terrestrial mosses, are described. Samples of different mosses, mainly Hypnum cupressiforme and Scleropodium purum, were collected from 134 sampling sites, between 1995 and 1996. Levels of Al, As, Cr, Cu, Fe, Hg, Ni, Pb and Zn, were determined by flame atomic absorption or atomic fluorescence spectrophotometry. Regression analysis was used to compare the capacity of the selected moss species to accumulate the elements, and intercalibration of accumulation in these species was carried out where necessary. Distribution maps were prepared to allow the zones most affected by metal deposition to be identified and to relate this to known sources of contamination: electricity power stations and other industries (e.g. Hg and Ni), edaphic contamination (e.g. Al and Cr) and road traffic (Pb). Background levels of metals in each species were also determined for the study area

Histological response of peritoneal carcinomatosis after hyperthermic intraperitoneal chemoperfusion (HIPEC) in experimental investigations

BACKGROUND: In selected patients with peritoneal carcinomatosis from colorectal cancer prognosis can be improved by hyperthermic intraperitoneal chemotherapy (HIPEC) after cytoreductive surgery. The aim of this study was to evaluate the tumor response of peritoneal carcinomatosis in tumor-bearing rats treated with HIPEC. METHODS: CC531 colon carcinoma (2,5 × 10(6 )cells), implanted intraperitoneally in Wag/Rija rats, was treated by hyperthermic intraperitoneal chemotherapy. After 10 days of tumor growth the animals were randomized into five groups of six animals each: group I: control (n = 6), group II: sham operated animals (n = 6), group III: hyperthermic intraperitoneal perfusion (HIP) without cytostatic drugs, group IV: HIPEC with mitomycin C in a concentration of 15 mg/m(2 )(n = 6), group V: mitomycin C i.p. alone in a concentration of 10 mg/m(2 )(n = 6). After 10 days the extent of tumor spread and histological outcome were analysed by autopsy. RESULTS: All control animals developed extensive intraperitoneal tumor growth. Histological tumor load was significantly reduced in group III and group V and was lowest in group IV. In group II tumor load was significantly higher than in group I. Implanted metastases were significantly decreased in group IV compared with group I and group II. CONCLUSION: These findings indicate that HIPEC is an effective treatment for peritoneal carcinomatosis in this animal model. HIPEC reduced macroscopic and microscopic intraperitoneal tumor spread

An efficient urine peptidomics workflow identifies chemically defined dietary gluten peptides from patients with celiac disease

Celiac disease (CeD) is an autoimmune disorder induced by consuming gluten proteins from wheat, barley, and rye. Glutens resist gastrointestinal proteolysis, resulting in peptides that elicit inflammation in patients with CeD. Despite well-established connections between glutens and CeD, chemically defined, bioavailable peptides produced from dietary proteins have never been identified from humans in an unbiased manner. This is largely attributable to technical challenges, impeding our knowledge of potentially diverse peptide species that encounter the immune system. Here, we develop a liquid chromatographic-mass spectrometric workflow for untargeted sequence analysis of the urinary peptidome. We detect over 600 distinct dietary peptides, of which ~35% have a CeD-relevant T cell epitope and ~5% are known to stimulate innate immune responses. Remarkably, gluten peptides from patients with CeD qualitatively and quantitatively differ from controls. Our results provide a new foundation for understanding gluten immunogenicity, improving CeD management, and characterizing the dietary and urinary peptidomes.Ministerio de Ciencia e Innovación SAF2017-83700-

The model of dynamo with small number of modes and magnetic activity of T Tauri stars

The model that describes operation of dynamo in fully convective stars is presented. It is based on representation of stellar magnetic field as a superposition of finite number of poloidal and toroidal free damping modes. In the frame of adopted low of stellar differential rotation we estimated minimal value of dynamo number D, starting from which generation of cyclic magnetic field in stars without radiative core is possible. We also derived expression for period of the cycle. It was found that dynamo cycles of fully convective stars and stars with thin convective envelopes differ in a qualitative way: 1) distribution of spots over latitude during the cycle is different in these stars; 2) the model predicts that spot formation in fully convective stars should be strongly suppressed at some phases of the cycle. We have analyzed historical lightcurve of WTTS star V410 Tau and found that long term activity of the star is not periodic process. Rather one can speak about quasi cyclic activity with characteristic time of $\sim 4$ yr and chaotic component over imposed. We concluded also that redistribution of cool spots over longitude is the reason of long term variations of V410 Tau brightness. It means that one can not compare directly results of photometric observations with predictions of our axially symmetric (for simplicity) model which allows to investigate time evolution of spot's distribution over latitude. We then discuss what kind of observations and in which way could be used to check predictions of the dynamo theory.Comment: 18 pages, 5 figures, accepted to Astron. Let

Orally Bioavailable Androgen Receptor Degrader, Potential Next-Generation Therapeutic for Enzalutamide-Resistant Prostate Cancer

Acknowledgement. BGS acknowledges work performed at the Center for Nanophase Materials Sciences, a DOE Office of Science User Facility. VB acknowledges Laboratory Directed Research and Development program of Oak Ridge National Laboratory, managed by UTBattelle, LLC, for the U.S. Department of Energy.Peer reviewedPostprin

Quasars Probing Quasars III: New Clues to Feedback, Quenching, and the Physics of Massive Galaxy Formation

Galaxies hosting z~2 quasars are the high-$z$ progenitors of today's massive `red-and-dead' galaxies. With close pairs of quasars at different redshifts, a background quasar can be used to study a foreground quasar's halo gas in absorption, providing a wealth of information about feedback, quenching, and the physics of massive galaxy formation. We present a Keck/HIRES spectrum of the bright background quasar in a projected pair with angular separation 13.3" corresponding to 108kpc at the redshift of the foreground quasar z_fg=2.4360 +/- 0.0005, precisely determined from Gemini/GNIRS near-IR spectroscopy. Our echelle spectrum reveals optically thick gas (NHI~10^19.7), coincident with the foreground quasar redshift. The ionic transitions of associated metal-lines reveal the following properties of the foreground quasar's halo: (1) the kinematics are extreme with absorption extending to +780km/s relative to z_fg; (2) the metallicity is nearly solar; (3) the temperature of the predominantly ionized gas is T<~20,000K; (4) the electron density is n_e~1 cm^-3 indicating a characteristic size ~10 - 100pc for the absorbing `clouds'; (7) there is a negligible amount of warm gas 10^5K < T < 10^6K; (8) the gas is unlikely illuminated by the foreground quasar, implying anisotropic or intermittent emission. The mass of cold T~10^4K gas implied by our observations is significant, amounting to a few percent of the total expected baryonic mass density of the foreground quasar's dark halo at r~100kpc. The origin of this material is still unclear, and we discuss several possibilities in the context of current models of feedback and massive galaxy formation.Comment: Accepted to ApJ. 38 pages, 11 figure