The Antiglucocorticoid RU486 Inhibits Phenobarbital Induction of the Chicken CYP2H1 Gene in Primary Hepatocytes

ABSTRACT The cytochrome P450 gene CYP2H1 is highly induced by phenobarbital in chick embryo hepatocytes. Recent studies have established that the orphan nuclear receptor CAR plays a critical role in the induction mechanism. Here, we show that a high concentration of the potent glucocorticoid and progesterone receptor antagonist RU486 almost completely blocks phenobarbital-induced accumulation of CYP2H1 mRNA in hepatocytes yet has no effect on basal expression. In marked contrast, CYP2H1 mRNA induced by the phenobarbital-type inducers glutethimide and 2-allylisopropylacetamide is not affected by RU486. RU486 inhibition is not mediated through the glucocorticoid or progesterone receptors. Transient transfection studies showed that RU486 does not repress through activation of the orphan receptor PXR and subsequent competition with CAR for binding to the upstream drug-responsive 556-base-pair enhancer. Additionally, none of the known functional transcription factor binding sites found in the enhancer region was a target of RU486 inhibition. Using an artificial construct containing multiple CAR binding sites, we also established that RU486 has no direct effect on the activity of exogenously expressed CAR. There is no evidence that phenobarbital binds to CAR; we propose that RU486 inhibits phenobarbital induction, either by interfering with a phenobarbital-dependent mechanism responsible for nuclear import of CAR or with the metabolism of phenobarbital to the true inducer. Whether a novel nuclear receptor that binds RU486 at high concentrations plays a role in the inhibitory action of RU486 is an interesting possibility

Ballistic protective properties of material representative of English civil war buff-coats and clothing

One type of clothing system used in the English Civil War, more common amongst cavalrymen than infantrymen, was the linen shirt, wool waistcoat and buff-coat. Ballistic testing was conducted to estimate the velocity at which 50% of 12-bore lead spherical projectiles (V50) would be expected to perforate this clothing system when mounted on gelatine (a tissue simulant used in wound ballistic studies). An estimated six-shot V50 for the clothing system was calculated as 102 m/s. The distance at which the projectile would have decelerated from the muzzle of the weapon to this velocity in free flight was triple the recognised effective range of weapons of the era suggesting that the clothing system would provide limited protection for the wearer. The estimated V50 was also compared with recorded bounce-and-roll data; this suggested that the clothing system could provide some protection to the wearer from ricochets. Finally, potential wounding behind the clothing system was investigated; the results compared favourably with seventeenth century medical writings

Mult Scler

Background: Investigating the degeneration of specific thalamic nuclei in multiple sclerosis (MS) remains challenging. Methods: White-matter-nulled (WMn) MPRAGE, MP-FLAIR, and standard T1-weighted magnetic resonance imaging (MRI) were performed on MS patients (n = 15) and matched controls (n = 12). Thalamic lesions were counted in individual sequences and lesion contrast-to-noise ratio (CNR) was measured. Volumes of 12 thalamic nuclei were measured using an automatic segmentation pipeline specifically developed for WMn-MPRAGE. Results: WMn-MPRAGE showed more thalamic MS lesions (n = 35 in 9 out of 15 patients) than MP-FLAIR (n = 25) and standard T1 (n = 23), which was associated with significant improvement of CNR (p < 0.0001). MS patients had whole thalamus atrophy (p = 0.003) with lower volumes found for the anteroventral (p < 0.001), the pulvinar (p < 0.0001), and the habenular (p = 0.004) nuclei. Conclusion: WMn-MPRAGE and automatic thalamic segmentation can highlight thalamic MS lesions and measure patterns of focal thalamic atrophy. © The Author(s), 2019.Translational Research and Advanced Imaging LaboratoryBordeaux Region Aquitaine Initiative for Neuroscienc

Investigation of transverse collective flow of intermediate mass fragments

The transverse flow of intermediate mass fragments (IMFs) has been investigated for the 35 MeV/u ${}^{70}\mathrm{Zn}+{}^{70}\mathrm{Zn}$, ${}^{64}\mathrm{Zn}+{}^{64}\mathrm{Zn}$, and ${}^{64}\mathrm{Ni}+{}^{64}\mathrm{Ni}$ systems. A transition from the IMF transverse flow strongly depending on the mass of the system, in the most violent collisions, to a dependence on the charge of the system, for the peripheral reactions, is shown. This transition was shown to be sensitive to the density dependence of the symmetry energy using the antisymmetrized molecular-dynamics model. The results present an observable, the IMF transverse flow, that can be used to probe the nuclear equation of state. Comparison with the simulation demonstrated a preference for a stiff density dependence of the symmetry energy

Effect of ketoconazole on the pharmacokinetics of axitinib in healthy volunteers

Objective Axitinib (AG-013736), an oral, potent, and selective inhibitor of vascular endothelial growth factor receptors 1, 2, and 3, is metabolized primarily by cytochrome P450 (CYP) 3A with minor contributions from CYP1A2, CYP2C19, and glucuronidation. Co-administration with CYP inhibitors may increase systemic exposure to axitinib and alter its safety profile. This study evaluated changes in axitinib plasma pharmacokinetic parameters and assessed safety and tolerability in healthy subjects, following axitinib co-administration with the potent CYP3A inhibitor ketoconazole. Methods In this randomized, single-blind, two-way crossover study, 32 healthy volunteers received placebo, followed by a single 5-mg oral dose of axitinib, administered either alone or on the fourth day of dosing with oral ketoconazole (400 mg/day for 7 days). Results Axitinib exposure was significantly increased in the presence of ketoconazole, with a geometric mean ratio for area under the plasma concentration–time curve from time zero to infinity of 2.06 (90% confidence interval [CI]: 1.84–2.30) and a geometric mean ratio for maximum plasma concentration (Cmax) of 1.50 (90% CI: 1.33–1.70). For axitinib alone or with ketoconazole, Cmax occurred 1.5 and 2.0 h after dosing, respectively. Adverse events were predominantly mild; the most commonly reported treatment-related adverse events were headache and nausea. Conclusions Axitinib plasma exposures and peak concentrations were increased following concurrent administration of axitinib and ketoconazole in healthy volunteers. Axitinib alone and in combination with ketoconazole was well tolerated. These findings provide an upper exposure for expected axitinib plasma concentrations in the presence of potent metabolic inhibition

Validity of oral mucosal transudate specimens for HIV testing using enzyme-linked immunosorbent assay in children in Chimanimani district, Zimbabwe

Objective. To assess the validity of oral mucosal transudate (OMT) specimens for HIV testing in children using enzyme-linked immunosorbent assay (ELISA). Methods. A cross-sectional descriptive study was conducted as part of a community-based behavioural and HIV sero-status survey of adults and children in the Chimanimani district of Zimbabwe. Dried blood spot (DBS) and OMT samples were collected from children aged between 2 and 14 years, inclusive. Both samples were tested for HIV using the Vironostika Uniform II plus O kits. The main study outcomes were the sensitivity and specificity of OMT samples, with DBS as the gold-standard specimen. Results. Paired DBS and OMT specimens were available from 1 274 (94.4%) of the 1 350 children enrolled. Using the DBS, HIV prevalence was 3.2%. Overall sensitivity of OMT was 48.8% (95% confidence interval (CI) 33.3 - 64.5), and specificity was 98.5% (95% CI 97.7 - 99.1). Conclusion. The overall sensitivity of OMT specimens for HIV testing in children using ELISA was low. Stratifying the analysis by sector showed that OMT samples are good specimens for HIV testing. It is important to note that factors such as the low HIV prevalence in our study population, quality of the OMT, diet and oral hygiene could have influenced the results

Sensitivity of intermediate mass fragment flows to the symmetry energy

The NIMROD-ISiS array was used to study the transverse flow of intermediate mass fragments in 35 MeV/nucleon ${}^{70}\mathrm{Zn}+{}^{70}\mathrm{Zn}$, ${}^{64}\mathrm{Zn}+{}^{64}\mathrm{Zn}$, and ${}^{64}\mathrm{Ni}+{}^{64}\mathrm{Ni}$ reactions. The intermediate mass fragment flow was previously shown to be sensitive to the density dependence of the symmetry energy. To explore the model dependence of the results, the antisymmetrized molecular dynamics, constrained molecular dynamics, and stochastic mean-field models were each compared to the experimental results to extract information on the form of the symmetry energy. The results demonstrate that sensitivity of the models to the nuclear equation of state can vary significantly based on the treatment of the nuclear dynamics. Despite the differences in the sensitivity, improved agreement with the experimental data is observed for each model with a stiff density dependence of the symmetry energy

Transverse collective flow and midrapidity emission of isotopically identified light charged particles

The transverse flow and relative midrapidity yield of isotopically identified light charged particles (LCPs) has been examined for the 35 MeV/nucleon ${}^{70}\mathrm{Zn}+{}^{70}\mathrm{Zn}$, ${}^{64}\mathrm{Zn}+{}^{64}\mathrm{Zn}$, and ${}^{64}\mathrm{Ni}+{}^{64}\mathrm{Ni}$ systems. A large enhancement of the midrapidity yield of the LCPs was observed relative to the yield near the projectile rapidity. In particular, this enhancement was increased for the more neutron-rich LCPs demonstrating a preference for the production of neutron-rich fragments in the midrapidity region. Additionally, the transverse flow of the LCPs was extracted, which provides insight into the average movement of the particles in the midrapidity region. Isotopic and isobaric effects were observed in the transverse flow of the fragments. In both cases, the transverse flow was shown to decrease with an increasing neutron content in the fragments. A clear inverse relationship between the transverse flow and the relative midrapidity yield is shown. The increased relative midrapidity emission produces a decreased transverse flow. The stochastic mean-field model was used for comparison to the experimental data. The results showed that the model was able to reproduce the general isotopic and isobaric trends for the midrapidity emission and transverse flow. The sensitivity of these observables to the density dependence of the symmetry energy was explored. The results indicate that the transverse flow and midrapidity emission of the LCPs are sensitive to the denisty dependence of the symmetry energy

Single-cell RNA-seq reveals dynamic paracrine control of cellular variation

High-throughput single-cell transcriptomics offers an unbiased approach for understanding the extent, basis and function of gene expression variation between seemingly identical cells. Here we sequence single-cell RNA-seq libraries prepared from over 1,700 primary mouse bone-marrow-derived dendritic cells spanning several experimental conditions. We find substantial variation between identically stimulated dendritic cells, in both the fraction of cells detectably expressing a given messenger RNA and the transcript’s level within expressing cells. Distinct gene modules are characterized by different temporal heterogeneity profiles. In particular, a ‘core’ module of antiviral genes is expressed very early by a few ‘precocious’ cells in response to uniform stimulation with a pathogenic component, but is later activated in all cells. By stimulating cells individually in sealed microfluidic chambers, analysing dendritic cells from knockout mice, and modulating secretion and extracellular signalling, we show that this response is coordinated by interferon-mediated paracrine signalling from these precocious cells. Notably, preventing cell-to-cell communication also substantially reduces variability between cells in the expression of an early-induced ‘peaked’ inflammatory module, suggesting that paracrine signalling additionally represses part of the inflammatory program. Our study highlights the importance of cell-to-cell communication in controlling cellular heterogeneity and reveals general strategies that multicellular populations can use to establish complex dynamic responses.National Human Genome Research Institute (U.S.). Centers of Excellence in Genomic Science (1P50HG006193-01)National Institutes of Health (U.S.). Pioneer Award (DP1OD003958-01)Howard Hughes Medical InstituteBroad Institute of MIT and Harvard. Klarman Cell Observator