Destabilizing resonances of precessing inertia-gravity waves.

Instabilities in stratified precessing fluid are investigated. We extend the study by Mahalov [Phys. Fluids A 5, 891 (1993)0899-821310.1063/1.858635] in the stably stratified Boussinesq framework, with an external Coriolis force (with rate Ω_{p}) altering the base flow through the distortion of the circular streamlines of the unperturbed axially stratified rotating columns (with constant vorticity 2Ω.) It is shown that the inviscid part of the modified velocity flow (0,Ωr,-2ɛΩrsinφ) and buoyancy with gradient N^{2}(-2ɛcosφ,2ɛsinφ,1) are an exact solution of Boussinesq-Euler equations. Here (r,φ,z) is a cylindrical coordinate system, with ɛ=Ω_{p}/Ω being the Poincaré number and N the Brunt-Väisälä frequency. The base flow is transformed into a Cartesian coordinate system, and the stability of a superimposed perturbation is studied in terms of Fourier (or Kelvin) modes. The resulting Floquet system for the Fourier modes has three parameters: ɛ, N=N/Ω, and µ, which is the angle between the wave vector k and the solid-body rotation axis in the limit ɛ=0. In this limit, there are inertia-gravity waves propagating with frequency ±ω and the resonant cases are those for which 2ω=nΩ, n being an integer. We perform an asymptotic analysis to leading order in ɛ and characterize the destabilizing resonant case of order n=1 (i.e., the subharmonic instability) which exists and for 0≤N<Ω/2. In this range, the subharmonic instability remains the strongest with a maximal growth rate σ_{m}=[ɛ(5sqrt[15]/8)sqrt[1-4N^{2}]/(4-N^{2})]. Stable stratification acts in such a way as to make the subharmonic instability less efficient, so as it disappears for N≥0.5Ω. The destabilizing resonant cases of order n=2,3,4,5 are investigated in detail by numerical computations. The effect of viscosity on these instabilities is briefly addressed assuming the diffusive coefficients (kinematic and thermal) are equal. Likewise, we briefly investigate the case where N^{2}<0 and show that the instability associated to the mode with k_{3}=0 is the strongest.

Spectral energy scaling in precessing turbulence.

We study precessing turbulence, which appears in several geophysical and astrophysical systems, by direct numerical simulations of homogeneous turbulence where precessional instability is triggered due to the imposed background flow. We show that the time development of kinetic energy K occurs in two main phases associated with different flow topologies: (i) an exponential growth characterizing three-dimensional turbulence dynamics and (ii) nonlinear saturation during which K remains almost time independent, the flow becoming quasi-two-dimensional. The latter stage, wherein the development of K remains insensitive to the initial state, shares an important common feature with other quasi-two-dimensional rotating flows such as rotating Rayleigh-Bénard convection, or the large atmospheric scales: in the plane k_{∥}=0, i.e., the plane associated to an infinite wavelength in the direction parallel to the principal rotation axis, the kinetic energy spectrum scales as k_{⊥}^{-3}. We show that this power law is observed for wave numbers ranging between the Zeman "precessional" and "rotational" scales, k_{S}^{-1} and k_{Ω}^{-1}, respectively, at which the associated background shear or inertial timescales are equal to the eddy turnover time. In addition, an inverse cascade develops for (k_{⊥},k)

Energy partition, scale by scale, in magnetic Archimedes Coriolis weak wave turbulence.

Magnetic Archimedes Coriolis (MAC) waves are omnipresent in several geophysical and astrophysical flows such as the solar tachocline. In the present study, we use linear spectral theory (LST) and investigate the energy partition, scale by scale, in MAC weak wave turbulence for a Boussinesq fluid. At the scale k^{-1}, the maximal frequencies of magnetic (Alfvén) waves, gravity (Archimedes) waves, and inertial (Coriolis) waves are, respectively, V_{A}k,N, and f. By using the induction potential scalar, which is a Lagrangian invariant for a diffusionless Boussinesq fluid [Salhi et al., Phys. Rev. E 85, 026301 (2012)PLEEE81539-375510.1103/PhysRevE.85.026301], we derive a dispersion relation for the three-dimensional MAC waves, generalizing previous ones including that of f-plane MHD "shallow water" waves [Schecter et al., Astrophys. J. 551, L185 (2001)AJLEEY0004-637X10.1086/320027]. A solution for the Fourier amplitude of perturbation fields (velocity, magnetic field, and density) is derived analytically considering a diffusive fluid for which both the magnetic and thermal Prandtl numbers are one. The radial spectrum of kinetic, S_{κ}(k,t), magnetic, S_{m}(k,t), and potential, S_{p}(k,t), energies is determined considering initial isotropic conditions. For magnetic Coriolis (MC) weak wave turbulence, it is shown that, at large scales such that V_{A}k/f≪1, the Alfvén ratio S_{κ}(k,t)/S_{m}(k,t) behaves like k^{-2} if the rotation axis is aligned with the magnetic field, in agreement with previous direct numerical simulations [Favier et al., Geophys. Astrophys. Fluid Dyn. (2012)] and like k^{-1} if the rotation axis is perpendicular to the magnetic field. At small scales, such that V_{A}k/f≫1, there is an equipartition of energy between magnetic and kinetic components. For magnetic Archimedes weak wave turbulence, it is demonstrated that, at large scales, such that (V_{A}k/N≪1), there is an equipartition of energy between magnetic and potential components, while at small scales (V_{A}k/N≫1), the ratio S_{p}(k,t)/S_{κ}(k,t) behaves like k^{-1} and S_{κ}(k,t)/S_{m}(k,t)=1. Also, for MAC weak wave turbulence, it is shown that, at small scales (V_{A}k/sqrt[N^{2}+f^{2}]≫1), the ratio S_{p}(k,t)/S_{κ}(t) behaves like k^{-1} and S_{κ}(k,t)/S_{m}(k,t)=1.

Cross-helicity in rotating homogeneous shear-stratified turbulence.

We consider homogeneous shear-stratified turbulence in a rotating frame, that exhibits complex nonlinear dynamics. Since the analysis of relative orientation between coupled fluctuating fields helps us to understand turbulence dynamics, we focus on the alignment properties of both the velocity and gravity fields with the potential vorticity gradient. With the help of statistical mechanics, we define a vector field which plays a role in the analogous so-called cross-helicity in magnetohydrodynamics. High-resolution direct numerical simulations of developed homogeneous baroclinic turbulence are performed, and a detailed analysis of probability density functions for cross-helicity is provided. A net preference for positive cross-helicity is shown to be related to a new alignment mechanism. We argue that the analysis of cross-helicity is crucial for understanding the dynamics of buoyancy driven flows.

Nonlinear dynamics and anisotropic structure of rotating sheared turbulence.

Homogeneous turbulence in rotating shear flows is studied by means of pseudospectral direct numerical simulation and analytical spectral linear theory (SLT). The ratio of the Coriolis parameter to shear rate is varied over a wide range by changing the rotation strength, while a constant moderate shear rate is used to enable significant contributions to the nonlinear interscale energy transfer and to the nonlinear intercomponental redistribution terms. In the destabilized and neutral cases, in the sense of kinetic energy evolution, nonlinearity cannot saturate the growth of the largest scales. It permits the smallest scale to stabilize by a scale-by-scale quasibalance between the nonlinear energy transfer and the dissipation spectrum. In the stabilized cases, the role of rotation is mainly nonlinear, and interacting inertial waves can affect almost all scales as in purely rotating flows. In order to isolate the nonlinear effect of rotation, the two-dimensional manifold with vanishing spanwise wave number is revisited and both two-component spectra and single-point two-dimensional energy components exhibit an important effect of rotation, whereas the SLT as well as the purely two-dimensional nonlinear analysis are unaffected by rotation as stated by the Proudman theorem. The other two-dimensional manifold with vanishing streamwise wave number is analyzed with similar tools because it is essential for any shear flow. Finally, the spectral approach is used to disentangle, in an analytical way, the linear and nonlinear terms in the dynamical equations.

Magnetized stratified rotating shear waves.

We present a spectral linear analysis in terms of advected Fourier modes to describe the behavior of a fluid submitted to four constraints: shear (with rate S), rotation (with angular velocity Ω), stratification, and magnetic field within the linear spectral theory or the shearing box model in astrophysics. As a consequence of the fact that the base flow must be a solution of the Euler-Boussinesq equations, only radial and/or vertical density gradients can be taken into account. Ertel's theorem no longer is valid to show the conservation of potential vorticity, in the presence of the Lorentz force, but a similar theorem can be applied to a potential magnetic induction: The scalar product of the density gradient by the magnetic field is a Lagrangian invariant for an inviscid and nondiffusive fluid. The linear system with a minimal number of solenoidal components, two for both velocity and magnetic disturbance fields, is eventually expressed as a four-component inhomogeneous linear differential system in which the buoyancy scalar is a combination of solenoidal components (variables) and the (constant) potential magnetic induction. We study the stability of such a system for both an infinite streamwise wavelength (k(1) = 0, axisymmetric disturbances) and a finite one (k(1) ≠ 0, nonaxisymmetric disturbances). In the former case (k(1) = 0), we recover and extend previous results characterizing the magnetorotational instability (MRI) for combined effects of radial and vertical magnetic fields and combined effects of radial and vertical density gradients. We derive an expression for the MRI growth rate in terms of the stratification strength, which indicates that purely radial stratification can inhibit the MRI instability, while purely vertical stratification cannot completely suppress the MRI instability. In the case of nonaxisymmetric disturbances (k(1) ≠ 0), we only consider the effect of vertical stratification, and we use Levinson's theorem to demonstrate the stability of the solution at infinite vertical wavelength (k(3) = 0): There is an oscillatory behavior for τ > 1+|K(2)/k(1)|, where τ = St is a dimensionless time and K(2) is the radial component of the wave vector at τ = 0. The model is suitable to describe instabilities leading to turbulence by the bypass mechanism that can be relevant for the analysis of magnetized stratified Keplerian disks with a purely azimuthal field. For initial isotropic conditions, the time evolution of the spectral density of total energy (kinetic + magnetic + potential) is considered. At k(3) = 0, the vertical motion is purely oscillatory, and the sum of the vertical (kinetic + magnetic) energy plus the potential energy does not evolve with time and remains equal to its initial value. The horizontal motion can induce a rapid transient growth provided K(2)/k(1)>>1. This rapid growth is due to the aperiodic velocity vortex mode that behaves like K(h)/k(h) where k(h)(τ)=[k(1)(2) + (K(2) - k(1)τ)(2)](1/2) and K(h) =k(h)(0). After the leading phase (τ > K(2)/k(1)>>1), the horizontal magnetic energy and the horizontal kinetic energy exhibit a similar (oscillatory) behavior yielding a high level of total energy. The contribution to energies coming from the modes k(1) = 0 and k(3) = 0 is addressed by investigating the one-dimensional spectra for an initial Gaussian dense spectrum. For a magnetized Keplerian disk with a purely vertical field, it is found that an important contribution to magnetic and kinetic energies comes from the region near k(1) = 0. The limit at k(1) = 0 of the streamwise one-dimensional spectra of energies, or equivalently, the streamwise two-dimensional (2D) energy, is then computed. The comparison of the ratios of these 2D quantities with their three-dimensional counterparts provided by previous direct numerical simulations shows a quantitative agreement.

Magnetohydrodynamic instabilities in rotating and precessing sheared flows: an asymptotic analysis.

Linear magnetohydrodynamic instabilities are studied analytically in the case of unbounded inviscid and electrically conducting flows that are submitted to both rotation and precession with shear in an external magnetic field. For given rotation and precession the possible configurations of the shear and of the magnetic field and their interplay are imposed by the "admissibility" condition (i.e., the base flow must be a solution of the magnetohydrodynamic Euler equations): we show that an "admissible" basic magnetic field must align with the basic absolute vorticity. For these flows with elliptical streamlines due to precession we undertake an analytical stability analysis for the corresponding Floquet system, by using an asymptotic expansion into the small parameter ε (ratio of precession to rotation frequencies) by a method first developed in the magnetoelliptical instabilities study by Lebovitz and Zweibel [Astrophys. J. 609, 301 (2004)]10.1086/420972. The present stability analysis is performed into a suitable frame that is obtained by a systematic change of variables guided by symmetry and the existence of invariants of motion. The obtained Floquet system depends on three parameters: ε , η (ratio of the cyclotron frequency to the rotation frequency) and χ=cos α, with α being a characteristic angle which, for circular streamlines, ε=0, identifies with the angle between the wave vector and the axis of the solid body rotation. We look at the various (centrifugal or precessional) resonant couplings between the three present modes: hydrodynamical (inertial), magnetic (Alfvén), and mixed (magnetoinertial) modes by computing analytically to leading order in ε the instabilities by estimating their threshold, growth rate, and maximum growth rate and their bandwidths as functions of ε, η, and χ. We show that the subharmonic "magnetic" mode appears only for η>square root of 5/2 and at large η (>>1) the maximal growth rate of both the "hydrodynamic" and magnetic modes approaches ε/2, while the one of the subharmonic "mixed" mode approaches zero.

Stability of rotating stratified shear flow: an analytical study.

We study the stability problem of unbounded shear flow, with velocity U(i)=Sx(3)delta(i1), subjected to a uniform vertical density stratification, with Brunt-Väisälä frequency N, and system rotation of rate Omega about an axis aligned with the spanwise (x(2)) direction. The evolution of plane-wave disturbances in this shear flow is governed by a nonhomogeneous second-order differential equation with time-dependent coefficients. An analytical solution is found to be described by Legendre functions in terms of the nondimensional parameter sigma(phi)(2)=R(R+1)sin(2) phi+R(i), where R=(2Omega/S) is the rotation number, phi is the angle between the horizontal wave vector and the streamwise axis, and R(i)=N(2)/S(2) is the Richardson number. The long-time behavior of the solution is analyzed using the asymptotic representations of the Legendre functions. On the one hand, linear stability is analyzed in terms of exponential growth, as in a normal-mode analysis: the rotating stratified shear flow is stable if R(i)>1/4, or if 00, or if R(R+1)<0

Precessing rotating flows with additional shear: stability analysis.

We consider unbounded precessing rotating flows in which vertical or horizontal shear is induced by the interaction between the solid-body rotation (with angular velocity Omega(0)) and the additional "precessing" Coriolis force (with angular velocity -epsilonOmega(0)), normal to it. A "weak" shear flow, with rate 2epsilon of the same order of the Poincaré "small" ratio epsilon , is needed for balancing the gyroscopic torque, so that the whole flow satisfies Euler's equations in the precessing frame (the so-called admissibility conditions). The base flow case with vertical shear (its cross-gradient direction is aligned with the main angular velocity) corresponds to Mahalov's [Phys. Fluids A 5, 891 (1993)] precessing infinite cylinder base flow (ignoring boundary conditions), while the base flow case with horizontal shear (its cross-gradient direction is normal to both main and precessing angular velocities) corresponds to the unbounded precessing rotating shear flow considered by Kerswell [Geophys. Astrophys. Fluid Dyn. 72, 107 (1993)]. We show that both these base flows satisfy the admissibility conditions and can support disturbances in terms of advected Fourier modes. Because the admissibility conditions cannot select one case with respect to the other, a more physical derivation is sought: Both flows are deduced from Poincaré's [Bull. Astron. 27, 321 (1910)] basic state of a precessing spheroidal container, in the limit of small epsilon . A Rapid distortion theory (RDT) type of stability analysis is then performed for the previously mentioned disturbances, for both base flows. The stability analysis of the Kerswell base flow, using Floquet's theory, is recovered, and its counterpart for the Mahalov base flow is presented. Typical growth rates are found to be the same for both flows at very small epsilon , but significant differences are obtained regarding growth rates and widths of instability bands, if larger epsilon values, up to 0.2, are considered. Finally, both flow cases are briefly discussed in view of a subsequent nonlinear study using pseudospectral direct numerical simulations, which is a natural continuation of RDT.

Time-dependent rotating stratified shear flow: exact solution and stability analysis.

A solution of the Euler equations with Boussinesq approximation is derived by considering unbounded flows subjected to spatially uniform density stratification and shear rate that are time dependent [S(t)= partial differentialU3/partial differentialx2]. In addition to vertical stratification with constant strength N(v)2, this base flow includes an additional, horizontal, density gradient characterized by N(h)2(t). The stability of this flow is then analyzed: When the vertical stratification is stabilizing, there is a simple harmonic motion of the horizontal stratification N(h)2(t) and of the shear rate S(t), but this flow is unstable to certain disturbances, which are amplified by a Floquet mechanism. This analysis may involve an additional Coriolis effect with Coriolis parameter f, so that governing dimensionless parameters are a modified Richardson number, R=[S(0)2+N(h)4(0)/N(v)2]1/2, and f(v)=f/N(v), as well as the initial phase of the periodic shear rate. Parametric resonance between the inertia-gravity waves and the oscillating shear is demonstrated from the dispersion relation in the limit R-->0. The parametric instability has connection with both baroclinic and elliptical flow instabilities, but can develop from a very different base flow.

Dopamine induces ERK activation in renal epithelial cells through H2O2 produced by monoamine oxidase.

BACKGROUND: The rat renal proximal tubule cells contain a large amount of monoamine oxidase, which catalyzes the oxidative deamination of catecholamines such as dopamine (DA). The aim of this study is to investigate the potential role of hydrogen peroxide (H2O2) produced by monoamine oxidase (MAO) isoform on regulation of cell signaling and function. METHODS: Primary rat proximal tubular cells, which contain almost exclusively MAO-A, and human embryonic kidney 293 (HEK 293) cells stably transfected with human MAO-B cDNA were treated with DA or tyramine in the presence or the absence of some inhibitors. Then, Shc protein tyrosine phosphorylation and extracellular-regulated kinase (ERK) activation were evaluated by immunoprecipitation/immunoblot analysis and cell proliferation by [3H]thymidine incorporation or cell counting. RESULTS: In rat proximal tubule cells, DA induced tyrosine phosphorylation of Shc, ERK activation, and a significant increase in DNA synthesis. The involvement of MAO-dependent H2O2 generation induced by DA (5 micromol/L) was supported by the demonstration that the DA effects were (1) fully prevented by cell pretreatment with the MAO inhibitor pargyline, the antioxydant N-acetylcysteine (NAC), and the DA uptake inhibitor GBR 12909; (2) not abrogated by the D1 and D2 receptor antagonists; (3) observed in HEK 293 MAO-B cells but not in HEK 293 wild-type cells, which do not express MAO; and (4) similar to those induced by another MAO substrate, tyramine. CONCLUSIONS: Taken together, these results show that in addition to the effects related to receptor stimulation, DA, and probably the other catecholamines, may induce some of its effects through the MAO-dependent H2O2 production.

Hydrogen peroxide generation by monoamine oxidases in rat white adipocytes: role on cAMP production.

In rat, white adipocytes monoamine oxidases (EC 1.4.3.4.) generate hydrogen peroxide (H(2)O(2)). Recent studies suggested that, in addition to its toxic features, H(2)O(2) may behave as a cell second messenger. In the present study, using fluorimetric and chemiluminescence (CL) assays, we showed that tyramine degradation by monoamine oxidases in intact adipocytes resulted in the concentration-dependent generation of H(2)O(2). In addition, we found that, in the presence of low tyramine concentrations, forskolin-dependent cAMP production was significantly increased as compared to that of the control and this increase was prevented by the monoamine oxidase inhibitor pargyline or by the H(2)O(2) trapping system homovanillic acid-peroxidase. Finally, we demonstrated that tyramine degradation by monoamine oxidases increased the ability of isoproterenol to induce cell lipolysis. Taken together, these data suggest that H(2)O(2) produced during substrate degradation by monoamine oxidases may participate in the regulation of adipocyte metabolism.

Monoamine oxidase B induces ERK-dependent cell mitogenesis by hydrogen peroxide generation.

The mitochondrial enzyme monoamine oxidase (MAO) A and B catalyze the oxidative deamination of various endogenous and exogenous biogenic amines. In the present study, we used human embryonic kidney 293 (HEK 293) cells stably transfected with human MAO-B cDNA to investigate the potential role of hydrogen peroxide (H(2)O(2)) produced by MAO-B isoform as an intracellular messenger involved in regulation of cell signaling and function. The MAO substrate tyramine induced tyrosine phosphorylation of Shc, ERK activation, and an increase in DNA synthesis in HEK 293 expressing MAO-B, but not in wild type HEK 293 cells, which do not express MAO. Tyramine effects were fully prevented by cell pretreatment with the MAO inhibitor pargyline or the antioxidant N-acetylcysteine. These results show that MAO-B induces MAPK/ERK activation and cell mitogenesis through H(2)O(2) production.

Reactive oxygen species production by monoamine oxidases in intact cells.

Monoamine oxidase (MAO) A and B are mitochondrial enzymes involved in the oxidative deamination of endogenous and exogenous amines. At present, the production of H2O2 by MAO in intact cells and its functional consequences in cell function have not been extensively investigated. The aim of this study was to define whether, in intact cells, the metabolism of small amounts of MAO substrates was able to induce a detectable H2O2 production. Hydrogen peroxide production was measured using a luminol-amplified chemiluminescence assay in three cell types, rat mesangial cells, rabbit proximal tubule cells and Hep-G2 cells, containing different MAO A/MAO B ratios. Our results showed that cell incubation with tyramine (50 micromol/l) led to a time-dependent H2O2 generation which was fully inhibited by MAO A (clorgyline and RO 41-1049) and MAO B (selegiline and RO 19-6327) inhibitors. The extent of inhibition of H2O2 production by selective inhibitors was in agreement with the amount of MAO isoforms expressed in each cell type, as determined by Western blot analysis and enzyme assay. Altogether, these findings show that, in a normal cell environment, MAO can be a source of reactive oxygen species which could have a functional impact on cell functions. In addition, we propose the luminol-amplified chemiluminescence assay as a rapid and sensitive procedure to characterize the monoamine oxidase isoforms and their regulation in intact cells.

IL-13 induces tyrosine phosphorylation of phospholipase C gamma-1 following IRS-2 association in human monocytes: relationship with the inhibitory effect of IL-13 on ROI production.

Here we analysed the involvement of tyrosine phosphorylation in the regulation of the initial molecular events induced by IL-13 to modulate TPA-triggered reactive oxygen intermediates (ROI) production. Our data indicate that treatment of monocytes with a protein tyrosine kinase inhibitor (herbimycin A) prevents IL-13-induced cAMP accumulation and subsequent ROI inhibition. We have previously demonstrated that cAMP accumulation depends on inositol phosphates hydrolysis (InsPs) and intracellular Ca2+ mobilisation. The inhibition of InsPs and intracellular Ca2+ release by herbimycin A suggests a primary role of tyrosine kinases upstream PLC activation. We further specify that IL-13 stimulates PLC-gamma 1 and IRS-2 tyrosine phosphorylation in human monocytes. We demonstrate for the first time that IL-13 induces the association of IRS-2 with PLC-gamma 1. We proposed here that PLC-gamma 1 is a new candidate recruited by IRS-2.

Immunocytochemical characterization of lung macrophage surface phenotypes and expression of cytokines in acute experimental silicosis in mice.

The expression of the surface phenotypical profile and the cytokines TNF-alpha and IL-1beta from murine lung macrophages was studied in parenchymal lung tissue and bronchoalveolar fluid of mice, over a 2-week period, following a single intratracheal instillation of silica. The acute inflammatory reaction, confirmed by a significant augmentation of four times the control values of the number of macrophages recovered by lavage from experimental animals, was followed by organized granulomas in the interstitium. The immunohistochemical analysis of lung tissue sections after silica instillation demonstrated the increased alveolar and interstitial tissue expression of all surface antigens and cytokines studied, mainly Mac-1, F4/80 antigens, TNF-alpha and IL-1beta, which were occasionally observed in normal uninjected and saline-treated mice. These findings show that, after silica instillation, the expression of surface phenotypical markers of lung macrophages increased, and this change was concomitantly associated with an increased expression of the cytokines TNF-alpha and IL-1beta. These changes support the conclusion that an influx of the newly recruited and activated macrophage population, with a different phenotype, is induced by treatment during inflammation. The populational changes involve difference in functional activity and enhance TNF-alpha and IL-1beta expression. These cytokines, produced in the silicosis-induced inflammatory process, are associated with the development of fibrosis and may contribute to disease severity.

Reactive oxygen intermediates as regulators of TNF-alpha production in rat lung inflammation induced by silica.

Exposure to mineral dusts such as silica has been associated with progressive pulmonary inflammation and fibrosis. There is evidence that the release of reactive oxygen intermediates (ROI) and cytokines by alveolar macrophages (AM) is involved in lung injury associated with silica exposure. However, the chronology and relationship between these two mediators are poorly understood. In this study, an animal model of silicosis has been used, allowing simultaneous follow-up of lung histopathologic state, AM TNF-alpha production at the protein (biologic assay) and mRNA (reverse transcriptase-PCR) levels, and the release of ROI (luminol-dependent chemiluminescence), after bronchoalveolar lavages. In particular, it has been shown that intratracheal instillation of silica (50 mg/kg) in rats led to fibrosis characterized by cellular interstitial infiltrates with granulomas, and in AM, it led to 1) an early and continuous increase in 12-O-tetradecanoylphorbol-13-acetate- or zymosan-triggered ROI production (days 1, 3, 14, and 28 post-treatment), and 2) a rise of TNF-alpha mRNA expression and protein secretion on days 3 and 14. A free radical scavenger pretreatment (N-ter-butyl-alpha-phenylnitrone) reversed lung histopathologic changes and decreased AM ROI production and TNF-alpha expression at the level of mRNA. These findings suggest that ROI production is an important primary event determining the silica-induced inflammatory process. ROI may act in an autocrine or paracrine manner and regulate TNF-alpha production by a mechanism promoting gene expression. The critical role of this cytokine in the pathogenesis of silicosis was confirmed by anti-TNF-alpha Ab treatment.

Interleukin-13 inhibits protein kinase C-triggered respiratory burst in human monocytes. Role of calcium and cyclic AMP.

Interleukin-13 (IL-13), a novel cytokine produced by activated lymphocytes modulates some monocyte functions, but no data is available concerning the signal transduction pathway. We show here, the inhibitory effect of IL-13 on 12-O-tetradecanoylphorbol-13-acetate (TPA)-triggered reactive oxygen intermediate production in human monocytes and the signals involved in this response. Our results show that IL-13 produces rapid and transient phosphoinositide hydrolysis and intracellular Ca2+ mobilization. Furthermore, IL-13 induces intracellular cAMP accumulation through inositol 1,4,5-trisphosphate-dependent Ca2+ mobilization. Metabolic inhibitors were used to relate the first steps in signaling pathways to the inhibitory effect of IL-13 on TPA-triggered reactive oxygen intermediate production. Indeed, inhibitors of phospholipase C (neomycin), intracellular Ca2+ mobilization (8-[N,N-diethylamino]-octyl 3,4,5-trimethoxybenzoate hydrochloride), adenylate cyclase (delta 9-tetrahydrocannabinol), and protein kinase A (N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide) impair the IL-13 inhibitory response. Altogether these observations indicate that modulatory effect of IL-13 on the TPA-induced oxidative burst is the result of the intracellular cAMP accumulation through an inositol 1,4,5-trisphosphate-induced Ca2+ mobilization-dependent pathway.

Ca(2+)-dependent activation of phospholipases C and D from mouse peritoneal macrophages by a selective trigger of Ca2+ influx, gamma-hexachlorocyclohexane.

The gamma-isomer of hexachlorocyclohexane (gamma-HCCH), which displays structural homology with inositol, was found to induce an initial influx of Ca2+ in mouse peritoneal macrophages. This was responsible for Ca(2+)-induced Ca2+ release via inositol 1,4,5-trisphosphate produced by phospholipase C and resulted in a sustained increase of cytoplasmic free Ca2+ concentration ([Ca2+]i). Entry of Ca2+ evoked by gamma-HCCH also stimulated phospholipase D, as well as the generation of reactive oxygen species formed by NADPH oxidase. These data suggest that some isoform(s) of phospholipase C, and possibly phospholipase D, can be activated by strictly Ca(2+)-dependent mechanisms. They also describe a new experimental tool allowing to trigger a selective influx of Ca2+. gamma-HCCH could thus be used in further studies aimed to delineate the role of Ca2+ entry in the subsequent activation of other signalling pathways.

Arachidonic acid metabolism of rat peritoneal macrophages after passive sensitization and allergen challenge.

The aim of our work was to evaluate the effect of passive sensitization of rat peritoneal macrophages (treatment of cells by an anti-ovalbumin IgE-rich serum) on arachidonic acid (AA) metabolism and the impact of specific antigen (ovalbumin) on this process. Compared to a control treatment without serum, the atopic serum and a serum without IgE, used on [3H]AA-labeled macrophages, increased cyclooxygenase and lipoxygenase eicosanoid production. Sera, used prior to [3H]AA incorporation, induced a decrease of 3H-labeled membrane phospholipids and an increase of lipoxygenase metabolites in the [3H]AA incorporation medium. To establish if the serum-induced catabolism of AA differed according to whether it was externally added to the culture medium or incorporated into membrane phospholipids, we studied the eicosanoid secretion of [3H]AA-prelabeled macrophages, treated by the serum and incubated with [14C]AA. It was confirmed that phospholipid-incorporated AA was catabolised following the cyclooxygenase and lipoxygenase pathways and external AA preferentially following the lipoxygenase pathway. The allergen increased the eicosanoid formation of passively sensitized macrophages but not that of cells treated by the serum without IgE. Our data suggest that changes occurring in passive sensitization, on AA mobilization from membrane phospholipids and on AA catabolism, induced by the serum, are independent of IgE and must be taken into consideration on interpreting the allergen effect.