Uncoupling of ATP-Mediated Calcium Signaling and Dysregulated Interleukin-6 Secretion in Dendritic Cells by Nanomolar Thimerosal

Dendritic cells (DCs), a rare cell type widely distributed in the soma, are potent antigen-presenting cells that initiate primary immune responses. DCs rely on intracellular redox state and calcium (Ca(2+)) signals for proper development and function, but the relationship between these two signaling systems is unclear. Thimerosal (THI) is a mercurial used to preserve vaccines and consumer products, and is used experimentally to induce Ca(2+) release from microsomal stores. We tested adenosine triphosphate (ATP)-mediated Ca(2+) responses of DCs transiently exposed to nanomolar THI. Transcriptional and immunocytochemical analyses show that murine myeloid immature DCs (IDCs) and mature DCs (MDCs) express inositol 1,4,5-trisphosphate receptor (IP(3)R) and ryanodine receptor (RyR) Ca(2+) channels, known targets of THI. IDCs express the RyR1 isoform in a punctate distribution that is densest near plasma membranes and within dendritic processes, whereas IP(3)Rs are more generally distributed. RyR1 positively and negatively regulates purinergic signaling because ryanodine (Ry) blockade a) recruited 80% more ATP responders, b) shortened ATP-mediated Ca(2+) transients > 2-fold, and c) produced a delayed and persistent rise (≥ 2-fold) in baseline Ca(2+). THI (100 nM, 5 min) recruited more ATP responders, shortened the ATP-mediated Ca(2+) transient (≥ 1.4-fold), and produced a delayed rise (≥ 3-fold) in the Ca(2+) baseline, mimicking Ry. THI and Ry, in combination, produced additive effects leading to uncoupling of IP(3)R and RyR1 signals. THI altered ATP-mediated interleukin-6 secretion, initially enhancing the rate of cytokine secretion but suppressing cytokine secretion overall in DCs. DCs are exquisitely sensitive to THI, with one mechanism involving the uncoupling of positive and negative regulation of Ca(2+) signals contributed by RyR1

Para- and Ortho-Substitutions Are Key Determinants of Polybrominated Diphenyl Ether Activity toward Ryanodine Receptors and Neurotoxicity

BackgroundPolybrominated diphenyl ethers (PBDEs) are widely used flame retardants that bioaccumulate in human tissues. Their neurotoxicity involves dysregulation of calcium ion (Ca(2+))signaling; however, specific mechanisms have yet to be defined.ObjectiveWe aimed to define the structure-activity relationship (SAR) for PBDEs and their metabolites toward ryanodine receptors type 1 (RyR1) and type 2 (RyR2) and to determine whether it predicts neurotoxicity.MethodsWe analyzed [3H]ryanodine binding, microsomal Ca(2+) fluxes, cellular measurements of Ca(2+) homeostasis, and neurotoxicity to define mechanisms and specificity of PBDE-mediated Ca(2+) dysregulation.ResultsPBDEs possessing two ortho-bromine substituents and lacking at least one para-bromine substituent (e.g., BDE-49) activate RyR1 and RyR2 with greater efficacy than corresponding congeners with two para-bromine substitutions (e.g., BDE-47). Addition of a methoxy group in the free para position reduces the activity of parent PBDEs. The hydroxylated BDEs 6-OH-BDE-47 and 4´-OH-BDE-49 are biphasic RyR modulators. Pretreatment of HEK293 cells (derived from human embryonic kidney cells) expressing either RyR1 or RyR2 with BDE-49 (250 nM) sensitized Ca2+ flux triggered by RyR agonists, whereas BDE-47 (250 nM) had negligible activity. The divergent activity of BDE-49, BDE-47, and 6-OH-BDE-47 toward RyRs predicted neurotoxicity in cultures of cortical neurons.ConclusionsWe found that PBDEs are potent modulators of RyR1 and RyR2. A stringent SAR at the ortho and para position determined whether a congener enhanced, inhibited, or exerted nonmonotonic actions toward RyRs. These results identify a convergent molecular target of PBDEs previously identified for noncoplanar polychlorinated biphenyls (PCBs) that predicts their cellular neurotoxicity and therefore could be a useful tool in risk assessment of PBDEs and related compounds

Interaction of the magnetorotational instability with hydrodynamic turbulence in accretion disks

Accretion disks in which angular momentum transport is dominated by the magnetorotational instability (MRI) can also possess additional, purely hydrodynamic, drivers of turbulence. Even when the hydrodynamic processes, on their own, generate negligible levels of transport, they may still affect the evolution of the disk via their influence on the MRI. Here, we study the interaction between the MRI and hydrodynamic turbulence using local MRI simulations that include hydrodynamic forcing. As expected, we find that hydrodynamic forcing is generally negligible if it yields a saturated kinetic energy density that is small compared to the value generated by the MRI. For stronger hydrodynamic forcing levels, we find that hydrodynamic turbulence modifies transport, with the effect varying depending upon the spatial scale of hydrodynamic driving. Large scale forcing boosts transport by an amount that is approximately linear in the forcing strength, and leaves the character of the MRI (for example the ratio between Maxwell and Reynolds stresses) unchanged, up to the point at which the forced turbulence is an order of magnitude stronger than that generated by the MRI. Low amplitude small scale forcing may modestly suppress the MRI. We conclude that the impact of hydrodynamic turbulence on the MRI is generically ignorable in cases, such as convection, where the additional turbulence arises due to the accretion energy liberated by the MRI itself. Hydrodynamic turbulence may affect (and either enhance or suppress) the MRI if it is both strong, and driven by independent mechanisms such as self-gravity, supernovae, or solid-gas interactions in multiphase protoplanetary disks.Comment: ApJ, in pres

Ryanodine receptor and FK506 binding protein 1 in the Atlantic killifish (Fundulus heteroclitus) : a phylogenetic and population-based comparison

Author Posting. © The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Aquatic Toxicology 192 (2017): 105-115, doi:10.1016/j.aquatox.2017.09.002.Non-dioxin-like polychlorinated biphenyls (NDL PCBs) activate ryanodine receptors (RyR), microsomal Ca2+ channels of broad significance. Teleost fish may be important models for NDL PCB neurotoxicity, and we used sequencing databases to characterize teleost RyR and FK506 binding protein 12 or 12.6 kDa (genes FKBP1A; FKBP1B), which promote NDL PCB-triggered Ca2+ dysregulation. Particular focus was placed on describing genes in the Atlantic killifish (Fundulus heteroclitus) genome and searching available RNA-sequencing datasets for single nucleotide variants (SNV) between PCB tolerant killifish from New Bedford Harbor (NBH) versus sensitive killifish from Scorton Creek (SC), MA. Consistent with the teleost whole genome duplication (tWGD), killifish have six RyR genes, corresponding to a and b paralogs of mammalian RyR1, 2 and 3. The presence of six RyR genes was consistent in all teleosts investigated including zebrafish. Killifish have four FKBP1; one FKBP1b and three FKBP1a named FKBP1aa, FKBP1ab, likely from the tWGD and a single gene duplicate FKBP1a3 suggested to have arisen in Atherinomorphae. The RyR and FKBP1 genes displayed tissue and developmental stage-specific mRNA expression, and the previously uncharacterized RyR3, herein named RyR3b, and all FKBP1 genes were prominent in brain. We identified a SNV in RyR3b encoding missense mutation E1458D. In NBH killifish, 57% were heterozygous and 28% were homozygous for this SNV, whereas almost all SC killifish (94%) lacked the variant (n≥39 per population). The outlined sequence differences between mammalian and teleost RyR and FKBP1 together with outlined population differences in SNV frequency may contribute to our understanding of NDL PCB neurotoxicity.This research was supported by the KC Donnelly Research Externship made possible by the National Institute of Environmental Health Sciences’ Superfund Research Program (EBH) and the Superfund Research Programs at UC Davis (INP and EBH; P42ES004699) and Boston University (JJS, JVG, MEH, SIK; P42ES007381). Additional support was provided by the National Institute of Health (INP; R01 ES014901; and P01 AR052354) and by National Science Foundation collaborative research grants (MEH and SIK; DEB-1265282 and DEB-1120263). This research was also supported in part by an appointment (to BC) with the Postdoctoral Research Program at the U.S. Environmental Protection (US EPA) Office of Research and Development administered by the Oak Ridge Institute for Science and Education (ORISE) through Interagency Agreement No. DW92429801 between the U.S. Department of Energy and the US EPA

On Self-Sustained Dynamo Cycles in Accretion Discs

(abridged) MHD turbulence is known to exist in shearing boxes with either zero or nonzero net magnetic flux. However, the way turbulence survives in the zero-net-flux case is not explained by linear theory and appears as a purely numerical result. Aims: We look for a nonlinear mechanism able to explain the persistence of MHD turbulence in shearing boxes with zero net magnetic flux, and potentially leading to large-scale dynamo action. Method: Spectral nonlinear simulations of the magnetorotational instability are shown to exhibit a large-scale axisymmetric magnetic field, maintained for a few orbits. The generation process of this field is investigated using the results of the simulations and an inhomogeneous linear approach. Results: The mechanism by which turbulence is sustained in zero-net-flux shearing boxes is shown to be related to the existence of a large-scale azimuthal field, surviving for several orbits. In particular, it is shown that MHD turbulence in shearing boxes can be seen as a dynamo process coupled to a magnetorotational-type instability.Comment: 11 pages, 11 figures, Accepted in A&

Accretion Disks and Dynamos: Toward a Unified Mean Field Theory

Conversion of gravitational energy into radiation in accretion discs and the origin of large scale magnetic fields in astrophysical rotators have often been distinct topics of research. In semi-analytic work on both problems it has been useful to presume large scale symmetries, necessarily resulting in mean field theories. MHD turbulence makes the underlying systems locally asymmetric and nonlinear. Synergy between theory and simulations should aim for the development of practical mean field models that capture essential physics and can be used for observational modeling. Mean field dynamo (MFD) theory and alpha-viscosity accretion theory exemplify such ongoing pursuits. 21st century MFD theory has more nonlinear predictive power compared to 20th century MFD theory, whereas accretion theory is still in a 20th century state. In fact, insights from MFD theory are applicable to accretion theory and the two are artificially separated pieces of what should be a single theory. I discuss pieces of progress that provide clues toward a unified theory. A key concept is that large scale magnetic fields can be sustained via local or global magnetic helicity fluxes or via relaxation of small scale magnetic fluctuations, without the kinetic helicity driver of 20th century textbooks. These concepts may help explain the formation of large scale fields that supply non-local angular momentum transport via coronae and jets in a unified theory of accretion and dynamos. In diagnosing the role of helicities and helicity fluxes in disk simulations, each disk hemisphere should be studied separately to avoid being misled by cancelation that occurs as a result of reflection asymmetry. The fraction of helical field energy in disks is expected to be small compared to the total field in each hemisphere as a result of shear, but can still be essential for large scale dynamo action.Comment: For the Proceedings of the Third International Conference and Advanced School "Turbulent Mixing and Beyond," TMB-2011 held on 21 - 28 August 2011 at the Abdus Salam International Centre for Theoretical Physics, Trieste, http://users.ictp.it/~tmb/index2011.html Italy, To Appear in Physica Scripta (corrected small items to match version in print

GLOBAL SIMULATIONS OF PROTOPLANETARY DISKS WITH OHMIC RESISTIVITY AND AMBIPOLAR DIFFUSION

Protoplanetary disks are believed to accrete onto their central T Tauri star because of magnetic stresses. Recently published shearing box simulations indicate that Ohmic resistivity, ambipolar diffusion and the Hall effect all play important roles in disk evolution. In the presence of a vertical magnetic field, the disk remains laminar between 1-5au, and a magnetocentrifugal disk wind forms that provides an important mechanism for removing angular momentum. Questions remain, however, about the establishment of a true physical wind solution in the shearing box simulations because of the symmetries inherent in the local approximation. We present global MHD simulations of protoplanetary disks that include Ohmic resistivity and ambipolar diffusion, where the time-dependent gas-phase electron and ion fractions are computed under FUV and X-ray ionization with a simplified recombination chemistry. Our results show that the disk remains laminar, and that a physical wind solution arises naturally in global disk models. The wind is sufficiently efficient to explain the observed accretion rates. Furthermore, the ionization fraction at intermediate disk heights is large enough for magneto-rotational channel modes to grow and subsequently develop into belts of horizontal field. Depending on the ionization fraction, these can remain quasi-global, or break-up into discrete islands of coherent field polarity. The disk models we present here show a dramatic departure from our earlier models including Ohmic resistivity only. It will be important to examine how the Hall effect modifies the evolution, and to explore the influence this has on the observational appearance of such systems, and on planet formation and migration.Comment: 18 pages, 12 figures, accepted for publication in Ap

Pycnogenol® and Ginkgo biloba extract: effect on peroxynitrite-oxidized sarcoplasmic reticulum Ca2+-ATPase

The effect of two natural standardized plant extracts, Pycnogenol® and EGb 761, on sarcoplasmic reticulum Ca2+-ATPase (SERCA) activity and posttranslational modifications induced by peroxynitrite was investigated to assess their possible protective role. EGb 761 was found to have a protective effect on SERCA activity in the concentration range of 5–40 µg/ml. On the other hand, Pycnogenol® caused a decrease of SERCA activity at concentrations of 25 µg/ml. EGb 761 did not prevent protein carbonyl formation upon oxidation with peroxynitrite. On the contrary, Pycnogenol® at the concentrations of 5 and 10 µg/ml significantly decreased the level of protein carbonyls by 44% and 54%, respectively. Neither Pycnogenol® nor EGb 761 exerted a protective effect against thiol group oxidation.The plant extracts studied modulated peroxynitrite-injured SERCA activity by different ways and failed to correlate with posttranslational modifications. Their effect seems to be associated with their ability to change SERCA conformation rather than by their antioxidant capacity

Lack of Evidence for Neonatal Misoprostol Neurodevelopmental Toxicity in C57BL6/J Mice

Misoprostol is a synthetic analogue of prostaglandin E1 that is administered to women at high doses to induce uterine contractions for early pregnancy termination and at low doses to aid in cervical priming during labor. Because of the known teratogenic effects of misoprostol when given during gestation and its effects on axonal growth in vitro, we examined misoprostol for its potential as a neurodevelopmental toxicant when administered to neonatal C57BL6/J mice. Mice were injected subcutaneously (s.c.) with 0.4, 4 or 40 µg/kg misoprostol on postnatal day 7, the approximate developmental stage in mice of human birth, after which neonatal somatic growth, and sensory and motor system development were assessed. These doses were selected to span the range of human exposure used to induce labor. In addition, adult mice underwent a battery of behavioral tests relevant to neurodevelopmental disorders such as autism including tests for anxiety, stereotyped behaviors, social communication and interactions, and learning and memory. No significant effects of exposure were found for any measure of development or behavioral endpoints. In conclusion, the results of the present study in C57BL/6J mice do not provide support for neurodevelopmental toxicity after misoprostol administration approximating human doses and timed to coincide with the developmental stage of human birth