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1.
Phys Rev E ; 101(5-1): 053207, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32575181

RESUMO

We studied parallel propagating electromagnetic waves in a magnetized quantum electron plasma of finite temperature, as an extension of our previous study on a zero temperature plasma. We obtained simple analytic dispersion relations in the long wavelength limit that included the thermal effect as correction terms to the zero temperature results. As in the zero temperature case, the lower branch of the R wave showed significant damping and became ill-defined at short wavelengths. Quantum effects seemed to give qualitative changes, such as the appearance of anomalous dispersion regions, to the classical dispersion relations when v_{F}/v_{th}≤0.2 for a set of exemplary parameters of v_{F}=0.1c and ω_{ce}/ω_{pe}=0.05 was used. We also noted that introduction of the Planck constant in the quantum Vlasov equation changed the shape of the anomalous dispersion region qualitatively, by forming a normal dispersion region in the middle of the original single broad anomalous dispersion region.

2.
Phys Rev E ; 96(3-1): 032133, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29346953

RESUMO

The impact of quantum statistics on the many-body dynamics of a crystalline solid at finite temperatures containing an interstitial solute atom (ISA) is investigated. The Mori-Zwanzig theory allows the many-body dynamics of the crystal to be formulated and solved analytically within a pseudo-one-particle approach using the Langevin equation with a quantum fluctuation-dissipation relation (FDR) based on the Debye model. At the same time, the many-body dynamics is also directly solved numerically via the molecular dynamics approach with a Langevin heat bath based on the quantum FDR. Both the analytical and numerical results consistently show that below the Debye temperature of the host lattice, quantum statistics significantly impacts the ISA transport properties, resulting in major departures from both the Arrhenius law of diffusion and the Einstein-Smoluchowski relation between the mobility and diffusivity. Indeed, we found that below one-third of the Debye temperature, effects of vibrations on the quantum mobility and diffusivity are both orders-of-magnitude larger and practically temperature independent. We have shown that both effects have their physical origin in the athermal lattice vibrations derived from the phonon ground state. The foregoing theory is tested in quantum molecular dynamics calculation of mobility and diffusivity of interstitial helium in bcc W. In this case, the Arrhenius law is only valid in a narrow range between ∼300 and ∼700 K. The diffusivity becomes temperature independent on the low-temperature side while increasing linearly with temperature on the high-temperature side.

3.
Phys Rev E ; 94(3-1): 032104, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27739747

RESUMO

The average energy needed to create a magnon is high in ferromagnetic metals due to the high-strength spin stiffness, which results in strong quantization effects that could be important even at thousands of degrees. To take into account quantum statistics at such high temperatures, the associated effects of anharmonicity of the spin vibrations must be taken into account. In addition to the complex nature of such effects, anharmonicity also affects the occupation of the density of state of the vibration states in the context of quantum statistics. Thus, an unoccupied vibration state might become occupied when its spring stiffness is substantially reduced with anharmonicity. Combined effects of quantum statistics and anharmonicity are expected. In this regard, the thermodynamics of ferromagnetic metals are investigated in this paper through the example of bcc iron between 10 and 1400 K. Theoretical analysis and spin-lattice dynamic simulations are performed, through which the physics behind the complex and dramatic temperature dependence of the thermodynamic functions of bcc iron is understood.

4.
Mol Oral Microbiol ; 29(5): 233-43, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24903905

RESUMO

Nasopharyngeal carriage of Streptococcus pneumoniae (pneumococcus) plays an important role in the development of invasive diseases, and is also critically involved in setting up respiratory bacterial and viral infections. We previously reported that pneumococcus, one of the commonly carried bacteria in the nasopharynx, regulates non-typeable Haemophilus influenzae-induced inflammation by upregulating the expression of Toll-like receptor 2 (TLR2). However, the underlying molecular mechanisms by which TLR2 expression is regulated during pneumococcal infections have not yet been well characterized. TBX21 is an important transcription factor of adaptive immunity, but there is an increasing body of evidence pointing to a role in regulating innate immunity. The expression of TBX21 was reported in epithelial cells, but the expression and role of TBX21 in respiratory epithelium, especially for regulating TLR2, has not yet been studied. In this study, we found that pneumococcus upregulates TBX21 expression in the respiratory epithelium. The effect of pneumococcus on TBX21 expression was dependent on its cytoplasmic toxin, pneumolysin. In addition, epithelial TBX21 expression was not regulated by the gram-negative bacterium non-typeable Haemophilus influenzae, peptidoglycan or endotoxin. Deficiency of TBX21 in mice or knocking down TBX21 in epithelial cells suppressed pneumococcus-induced TLR2 expression, but not that of TLR4 or TLR9. These results indicate that the adaptive immune regulator TBX21 participates in regulating innate immune responses, through regulation of TLR2 expression during pneumococcal infections.


Assuntos
Imunidade Inata/imunologia , Infecções Pneumocócicas/imunologia , Proteínas com Domínio T/imunologia , Receptor 2 Toll-Like/imunologia , Imunidade Adaptativa/imunologia , Animais , Proteínas de Bactérias/imunologia , Técnicas de Cultura de Células , Células Cultivadas , Orelha Média/imunologia , Endotoxinas/imunologia , Células Epiteliais/imunologia , Técnicas de Silenciamento de Genes , Haemophilus influenzae/imunologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Otite Média/imunologia , Otite Média/microbiologia , Peptidoglicano/imunologia , Pneumonia Pneumocócica/imunologia , Alvéolos Pulmonares/imunologia , RNA Interferente Pequeno/genética , Mucosa Respiratória/imunologia , Estreptolisinas/imunologia , Receptor 4 Toll-Like/análise , Receptor Toll-Like 9/análise , Regulação para Cima
5.
Water Res ; 58: 122-31, 2014 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-24747143

RESUMO

An assessment of the occurrence and concentration of enteric viruses and coliphages was carried out in highly urbanized catchment waters in the tropical city-state of Singapore. Target enteric viruses in this study were noroviruses, adenoviruses, astroviruses and rotaviruses. In total, 65 water samples were collected from canals and the reservoir of the Marina catchment on a monthly basis over a period of a year. Quantitative PCR (qPCR) and single agar layer plaque assay (SAL) were used to enumerate target enteric viruses and coliphages in water samples, respectively. The most prevalent pathogen were noroviruses, detected in 37 samples (57%), particularly norovirus genogroup II (48%), with a mean concentration of 3.7 × 10(2) gene copies per liter. Rotavirus was the second most prevalent virus (40%) with a mean concentration of 2.5 × 10(2) GC/L. The mean concentrations of somatic and male-specific coliphages were 2.2 × 10(2) and 1.1 × 10(2) PFU/100 ml, respectively. The occurrence and concentration of each target virus and the ratio of somatic to male-specific coliphages varied at different sampling sites in the catchment. For sampling sites with higher frequency of occurrence and concentration of viruses, the ratio of somatic to male-specific coliphages was generally much lower than other sampling sites with lower incidences of enteric viruses. Overall, higher statistical correlation was observed between target enteric viruses than between enteric viruses and coliphages. However, male-specific coliphages were positively correlated with norovirus concentrations. A multi-level integrated surveillance system, which comprises the monitoring of bacterial indicators, coliphages and selected enteric viruses, could help to meet recreational and surface water quality criteria in a complex urbanized catchment.


Assuntos
Colífagos/isolamento & purificação , Enterovirus/isolamento & purificação , Água Doce/virologia , Microbiologia da Água , Adenoviridae/genética , Adenoviridae/isolamento & purificação , Colífagos/genética , Enterovirus/genética , Monitoramento Ambiental , Norovirus/genética , Norovirus/isolamento & purificação , Reação em Cadeia da Polimerase , Recreação , Rotavirus/genética , Rotavirus/isolamento & purificação , Estações do Ano , Singapura , Clima Tropical , Reforma Urbana , Qualidade da Água
6.
Sci Rep ; 4: 3946, 2014 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-24492764

RESUMO

Ferroelectric vortex domain structure which exists in low-dimensional ferroelectrics is being intensively researched for future applications in functional nanodevices. Here we demonstrate that adjusting surface charge screening in combination with temperature can provide an efficient way to gain control of vortex domain structure in ferroelectric nanodot. Systematical simulating experiments have been conducted to reveal the stability and evolution mechanisms of domain structure in ferroelectric nanodot under various conditions, including processes of cooling-down/heating-up under different surface charge screening conditions, and increasing/decreasing surface charge screening at different temperatures. Fruitful phase diagrams as functions of surface screening and temperature are presented, together with evolution paths of various domain patterns. Calculations discover up to 25 different kinds of domain patterns and 22 typical evolution paths of phase transitions. The fruitful controllability of vortex domain structure by surface charge screening in combination with temperature should shed light on prospective nanodevice applications of low-dimensional ferroelectric nanostructures.

7.
J Nanosci Nanotechnol ; 10(5): 3357-61, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20358956

RESUMO

Olivine structured LiMPO4 (M = Fe, Mn, Co) was synthesized by the solvothermal process using a polyol medium of diethylene glycol (DEG) without any further heating as a post procedure. The synthesized LiMPO4 nanoparticles exhibited an average length of 200 approximately 400 nm, width of 200 approximately 300 nm, and thickness of 20 approximately 50 nm, and with well dispersed plate and rod morphologies. The Rietveld-refined X-ray diffraction (XRD) pattern of LiFePO4 exhibited a good fit with an olivine structure (space group: Pnma) and no undesirable impurity phases. The LiFePO4 nanocrystals showed a reversible capacity of 167 mAh/g, equating to a utilization efficiency of 98% of its theoretical capacity, with a good cyclability over an extended range up to 40 cycles.

8.
J Nanosci Nanotechnol ; 10(5): 3416-9, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20358969

RESUMO

LiFePO4/Carbon nanocomposites with varying amounts of carbon were synthesized using polyol process without any further heating as a post step. The X-ray diffraction patterns of all samples were indexed on the basis of orthorhombic olivine-type structure. The field emission-TEM images of composites with the amount of 3 approximately 15 wt% carbon showed the average sizes of 45 approximately 80 nm with nanorod shape surrounded by carbon. The initial discharge curves of LiFePO4/Carbon show 166, 157, 142, and 119 mAh/g capacities at a current density of 0.1 mA/cm2 in the voltage range of 2.5-4.2 V, respectively, without capacity fading and excellent cycle retentions during the 20 cycles. Especially, the sample of amount of 3 wt% carbon shows the excellent high rate performances at 15 and 30 C rates with high capacity retention compared with previously reported results of high temperature processed results, due to the existence of conductive agents such as carbon which suppresses particle growth and exhibits improved electronic conductivity and lithium ion diffusivity, simultaneously.

9.
Phys Rev E Stat Nonlin Soft Matter Phys ; 82(3 Pt 1): 031111, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21230029

RESUMO

In molecular dynamics simulations, temperature is evaluated, via the equipartition principle, by computing the mean kinetic energy of atoms. There is no similar recipe yet for evaluating temperature of a dynamic system of interacting spins. By solving semiclassical Langevin spin-dynamics equations, and applying the fluctuation-dissipation theorem, we derive an equation for the temperature of a spin ensemble, expressed in terms of dynamic spin variables. The fact that definitions for the kinetic and spin temperatures are fully consistent is illustrated using large-scale spin dynamics and spin-lattice dynamics simulations.

10.
Phys Rev E Stat Nonlin Soft Matter Phys ; 79(4 Pt 2): 046703, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19518376

RESUMO

To control numerical errors accumulated over tens of millions of time steps during the integration of a set of highly coupled equations of motion is not a trivial task. In this paper, we propose a parallel algorithm for spin dynamics and the newly developed spin-lattice dynamics simulation [P. W. Ma, Phys. Rev. B 78, 024434 (2008)]. The algorithm is successfully tested in both types of dynamic calculations involving a million spins. It shows good stability and numerical accuracy over millions of time steps (approximately 1 ns) . The scheme is based on the second-order Suzuki-Trotter decomposition (STD). The usage can avoid numerical energy dissipation despite the trajectory and machine errors. The mathematical base of the symplecticity, for properly decomposed evolution operators, is presented. Due to the noncommutative nature of the spin in the present STD scheme, a unique parallel algorithm is needed. The efficiency and stability are tested. It can attain six to seven times speed up when eight threads are used. The run time per time step is linearly proportional to the system size.

11.
Nanotechnology ; 20(7): 075401, 2009 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-19417419

RESUMO

The effect of an applied mechanical stress on the tunneling conductance of a ferroelectric tunnel barrier between substrates and dissimilar electrodes is studied. Using a thermodynamic model and taking into account polarization charge screening in the electrodes and the near-surface inhomogeneous polarization distribution, the tunneling conductance is calculated as a function of the applied stress. It is found that reversing an applied stress can also change the tunnel barrier sufficiently to produce an effect similar to the giant electro-resistance one due to polarization reversal, which is particularly significant near the stress-dependent paraelectric/ferroelectric phase transition. Indeed, the sensitivity is adequate for high-sensitivity electronic and mechanical sensors, memories and other nanodevices.

12.
Nano Lett ; 8(10): 3131-6, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18781805

RESUMO

The behavior of an uniaxially pulse-loaded ferroelectric nanowire is simulated using a Landau-Ginzburg type thermodynamic model. Our results show that under a load of suitable magnitude and frequency, an appropriately dimensioned ferroelectric nanowire can produce a sizable alternating current voltage, sufficient for applications as a nanopower source for energy harvesting, or as an effective nanomechanical sensor.


Assuntos
Nanotecnologia/métodos , Nanofios/química , Condutividade Elétrica , Fontes de Energia Elétrica , Ferro/química , Modelos Estatísticos , Nanopartículas/química , Estresse Mecânico , Resistência à Tração , Termodinâmica
13.
Phys Rev Lett ; 93(24): 245502, 2004 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-15697823

RESUMO

Nanoindentation-induced interlayer bond switching and phase transformation in carbon nanotubes (CNTs) and graphite are simulated by molecular dynamics. Both graphite and CNTs experience a soft-to-hard phase transformation at room temperature at compressive stresses of 12 and 16 GPa, respectively. Further penetration leads to the formation of interlayer sp(3) bonds, which are reversible upon unloading if the compressive stress is under about 70 GPa, beyond which permanent interlayer sp(3) bonds form. During nanoindentation, the maximum nanohardness of graphite can reach 109 GPa, and CNTs 120 GPa, which is comparable to that of diamond.

14.
J Clin Periodontol ; 28(9): 886-90, 2001 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11493360

RESUMO

AIM: The aim of this study was to determine the prevalence and the structure of the leukotoxin promoter region of Actinobacillus actinomycetemcomitans in an ethnic Chinese population. METHOD: Subgingival plaque samples were collected from 42 patients with moderate to advanced periodontitis and 50 periodontally healthy patients. A. actinomycetemcomitans was detected directly from the crude subgingival plaque by PCR using leukotoxin gene specific primers. The presence of A. actinomycetemcomitans was determined by a single 285 bp PCR amplicon. RESULTS: A. actinomycetemcomitans was found to be present in the subgingival plaque of 68 out of a total of 92 patients examined (74%). 29 out of the 42 periodontitis patients tested were carriers of A. actinomycetemcomitans (69%). Among the periodontally healthy patients studied, 39 out of 50 subjects possessed the bacteria (78%). PCR analysis of the promoter region of the ltx operon revealed that none of the 42 moderate to advanced periodontitis patients examined harboured A. actinomycetemcomitans strains with the JP2-like promoter of the ltx operon, known to enhance leukotoxin expression. 2 out of the 27 advanced periodontitis patients clinically diagnosed as suffering from rapidly progressive periodontitis were found to be carriers of the mildly toxic strain of A. actinomycetemcomitans with the characteristic 652-like promoter. CONCLUSIONS: The high prevalence of A. actinomycetemcomitans, regardless of whether the subgingival samples were analysed from patients with healthy or diseased periodontium suggests that this bacterial species is part of the normal oral flora of ethnic Chinese. Our preliminary results also suggested that subjects who harboured the mildly toxic strain of A. actinomycetemcomitans were potentially susceptible to aggressive forms of periodontitis.


Assuntos
Infecções por Actinobacillus/epidemiologia , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Exotoxinas/genética , Periodontite/microbiologia , Infecções por Actinobacillus/microbiologia , Adulto , Aggregatibacter actinomycetemcomitans/genética , Aggregatibacter actinomycetemcomitans/patogenicidade , Estudos de Casos e Controles , China/etnologia , Placa Dentária/microbiologia , Feminino , Regulação Bacteriana da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Prevalência , Singapura/epidemiologia
15.
Biochem Biophys Res Commun ; 285(3): 825-9, 2001 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-11453667

RESUMO

Exposure of mammalian cells to ultraviolet (UV) light elicits a cellular response and also lead to apoptotic cell death. However, the role of Rac, a member of Rho family GTPases, in the UV-induced apoptosis has never been examined. In UV-irradiated Rat-2 fibroblasts, nuclear fragmentation began to be observed within 2 h and the total viability of Rat-2 cells were only about 15% at 6 h following by UV irradiation, whereas the total viability in Rat2-Rac(N17) cells stably expressing RacN17, a dominant negative Rac1 mutant, was almost close to 67%. Pretreatment with SB203580, a specific inhibitor of p38 kinase, likewise attenuated UV-induced cell death, but PD98059, a MEK inhibitor, did not. Thus, Rac1 and p38 kinase appear to be components in the apoptotic signaling pathway induced by UV irradiation in Rat-2 fibroblasts. In addition, our results show that p38 kinase stimulation by UV is dramatically inhibited by RacN17, suggesting that p38 kinase is situated downstream of Rac1 in the UV signaling to apoptosis.


Assuntos
Apoptose , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Raios Ultravioleta , Proteínas rac1 de Ligação ao GTP/metabolismo , Animais , Bisbenzimidazol , Linhagem Celular , Sobrevivência Celular/efeitos da radiação , Inibidores Enzimáticos/farmacologia , Fibroblastos/citologia , Corantes Fluorescentes , Genes Dominantes , Imidazóis/farmacologia , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Mutação , Fosforilação/efeitos dos fármacos , Piridinas/farmacologia , Ratos , Transfecção , Proteínas Quinases p38 Ativadas por Mitógeno , Proteínas rac1 de Ligação ao GTP/genética
16.
Kidney Int ; 59(5): 1695-705, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11318940

RESUMO

BACKGROUND: The alteration of renal cell growth is one of the early abnormalities in the diabetic nephropathy. However, the effects of high glucose and its action mechanism in renal proximal tubule cell (PTC) proliferation have not been elucidated. METHODS: The effects of 25 mmol/L glucose on cell proliferation, thymidine, and leucine incorporation, cell cycle, and lipid peroxide formation were examined in the primary cultured renal PTCs. RESULTS: Glucose 25 mmol/L inhibited [3H]-thymidine incorporation and decreased cell growth. However, it increased [3H]-leucine incorporation and protein content. Furthermore, 25 mmol/L glucose increased lipid peroxide formation. These effects of glucose were blocked by antioxidants, vitamin E, N-acetylcystein, or taurine. Staurosporine and H-7 totally blocked 25 mmol/L glucose-induced lipid peroxide formation and had an inhibitory effect on [3H]-thymidine incorporation. Indeed, 25 mmol/L glucose increased the translocation of protein kinase C (PKC) from cytosolic fraction to membrane fraction. In addition, high glucose increased the secretion of transforming growth factor-beta1 (TGF-beta 1) via the PKC-oxidative stress pathway, and TGF-beta 1 inhibited [3H]-thymidine incorporation in a dose-dependent manner. CONCLUSIONS: High glucose inhibits renal PTC proliferation via PKC, oxidative stress, and the TGF-beta 1 signaling pathway.


Assuntos
Glucose/farmacologia , Túbulos Renais Proximais/citologia , Túbulos Renais Proximais/efeitos dos fármacos , Acetilcisteína/farmacologia , Animais , Antioxidantes/farmacologia , Ciclo Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Nefropatias Diabéticas/etiologia , Nefropatias Diabéticas/metabolismo , Nefropatias Diabéticas/patologia , Túbulos Renais Proximais/metabolismo , Leucina/metabolismo , Peróxidos Lipídicos/biossíntese , Estresse Oxidativo/efeitos dos fármacos , Proteína Quinase C/metabolismo , Coelhos , Taurina/farmacologia , Timidina/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Vitamina E/farmacologia
17.
J Biol Chem ; 276(27): 24645-53, 2001 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-11323430

RESUMO

Activation of Ras signaling by growth factors has been associated with gene regulation and cell proliferation. Here we characterize the contributory role of cytosolic phospholipase A(2) in the oncogenic Ha-Ras(V12) signaling pathway leading to activation of c-fos serum response element (SRE) and transformation in Rat-2 fibroblasts. Using a c-fos SRE-luciferase reporter gene, we showed that the transactivation of SRE by Ha-Ras(V12) is mainly via a Rac-linked cascade, although the Raf-mitogen-activated protein kinase cascade is required for full activation. In addition, Ha-Ras(V12)-induced DNA synthesis was significantly attenuated by microinjection of recombinant Rac(N17), a dominant negative mutant of Rac1. To identify the mediators downstream of Rac in the Ha-Ras(V12) signaling, we investigated the involvement of cytosolic phospholipase A(2). Oncogenic Ha-Ras(V12)-induced SRE activation was significantly inhibited by either pretreatment with mepacrine, a phospholipase A(2) inhibitor, or cotransfection with the antisense oligonucleotide of cytosolic phospholipase A(2). We also found cytosolic phospholipase A(2) to be situated downstream of Ha-Ras(V12) in a signal pathway leading to transformation. Together, these results are indicative of mediatory roles of Rac and cytosolic phospholipase A(2) in the signaling pathway by which Ha-Ras(V12) transactivates c-fos SRE and transformation. Our findings point to cytosolic phospholipase A(2) as a novel potential target for suppressing oncogenic Ha-Ras(V12) signaling in the cell.


Assuntos
Genes ras/fisiologia , Fosfolipases A/metabolismo , Transdução de Sinais , Proteínas ras/fisiologia , Animais , Sequência de Bases , Replicação do DNA/efeitos dos fármacos , Proteínas de Ligação a DNA/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Genes Reporter , Luciferases/genética , Microinjeções , Dados de Sequência Molecular , Proteínas Nucleares/metabolismo , Oligonucleotídeos Antissenso/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Fosfolipases A2 , Quinacrina/farmacologia , Ratos , Fator de Resposta Sérica , Ativação Transcricional/efeitos dos fármacos , Transfecção , Proteínas rac1 de Ligação ao GTP/metabolismo
18.
J Biol Chem ; 275(41): 32357-62, 2000 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-10934206

RESUMO

Reactive oxygen species (ROS) are important regulatory molecules implicated in the signaling cascade triggered by tumor necrosis factor (TNF)-alpha, although the events through which TNF-alpha induces ROS generation are not yet well characterized. We therefore investigated selected candidates likely to mediate TNF-alpha-induced ROS generation. Consistent with the role of Rac in that process, stable expression of Rac(Asn-17), a dominant negative Rac1 mutant, completely blocked TNF-alpha-induced ROS generation. To understand better the mediators downstream of Rac, we investigated the involvement of cytosolic phospholipase A(2) (cPLA(2)) activation and metabolism of the resultant arachidonic acid (AA) by 5-lipoxygenase (5-LO). TNF-alpha-induced ROS generation was blocked by inhibition of cPLA(2) or 5-LO, but not cyclooxygenase, suggesting that TNF-alpha-induced ROS generation is dependent on synthesis of AA and its subsequent metabolism to leukotrienes. Consistent with that hypothesis, TNF-alpha Rac-dependently stimulated endogenous production of leukotriene B(4) (LTB(4)), while exogenous application of LTB(4) increased levels of ROS. In contrast, application of leukotrienes C(4), D(4), and E(4) or prostaglandin E(2) had little effect. Our findings suggest that LTB(4) production by 5-LO is situated downstream of the Rac-cPLA(2) cascade, and we conclude that Rac, cPLA(2), and LTB(4) play pivotal roles in the ROS-generating cascade triggered by TNF-alpha.


Assuntos
Citosol/enzimologia , Fosfolipases A/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Animais , Araquidonato 5-Lipoxigenase/metabolismo , Ácido Araquidônico/metabolismo , Ácidos Araquidônicos/farmacologia , Linhagem Celular , Proteínas de Ligação a DNA/fisiologia , Ativação Enzimática , Genes Dominantes/genética , Genes fos/genética , Indóis/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno , Leucotrieno B4/metabolismo , Leucotrieno B4/farmacologia , Inibidores de Lipoxigenase , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Mutação/genética , Proteínas Nucleares/fisiologia , Fosfatidilinositol 3-Quinases/metabolismo , Fosfolipases A/antagonistas & inibidores , Fosfolipases A2 , Ratos , Receptores do Leucotrieno B4/metabolismo , Elementos de Resposta/genética , Fator de Resposta Sérica , Proteínas rac1 de Ligação ao GTP/genética , Proteínas rac1 de Ligação ao GTP/metabolismo
19.
Biochem J ; 348 Pt 3: 525-30, 2000 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-10839982

RESUMO

Although there have been a number of recent studies on the role of Rac in the generation of reactive oxygen species (ROS), details of the signalling pathway remain unclear. In the present study we analysed the extent to which the activation of cytosolic phospholipase A(2) and the resultant release of arachidonic acid (AA) are involved in the Rac-mediated generation of ROS. Transfection of Rat-2 cells with RacV12, a constitutively active form of Rac1, induced elevated levels of ROS, as reflected by increased H(2)O(2)-sensitive fluorescence of 2', 7'-dichlorofluorescein. These effects could be blocked by inhibiting phospholipase A(2) or 5-lipoxygenase but not by inhibiting cyclo-oxygenase. The application of exogenous AA increased levels of ROS but the effect was dependent on the further metabolism of AA to leukotrienes C(4)/D(4)/E(4) by 5-lipoxygenase. Indeed, the exogenous application of a mixture of leukotrienes C(4)/D(4)/E(4) elicited transient elevations in the levels of ROS that were blocked by catalase. These findings indicate that phospholipase A(2) and subsequent AA metabolism by 5-lipoxygenase act as downstream mediators in a Rac signalling pathway leading to the generation of ROS.


Assuntos
Ácido Araquidônico/metabolismo , Citosol/enzimologia , Fosfolipases A/metabolismo , Espécies Reativas de Oxigênio , Transdução de Sinais , Proteínas rac de Ligação ao GTP/metabolismo , Animais , Transporte Biológico , Linhagem Celular , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Leucotrienos/farmacologia , Fosfolipases A2 , Ratos
20.
Biochem Biophys Res Commun ; 268(1): 231-6, 2000 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-10652241

RESUMO

Rac is an important regulatory molecule implicated in c-jun N-terminal kinase (JNK) activation in response to stress and cytokines. However, the signaling events that mediate the activation of JNK by Rac are not yet well characterized. To broaden our understanding of downstream mediators that link Rac signals to the JNK pathway, we investigated whether cytosolic phospholipase A(2) (cPLA(2)) is involved in Rac activation of JNK. In this report we demonstrate that either co-transfection with antisense cPLA(2) oligonucleotide or pretreatment with arachidonyltrifluoromethyl ketone (AACOCF3), a potent and specific inhibitor of cPLA(2), inhibits Rac-mediated JNK activation, implying a potential role of cPLA(2) in Rac-signaling to JNK activation. In accordance with this observation, we demonstrate that the addition of exogenous arachidonic acid (AA), a principal product of Rac-activated cPLA(2), or leukotrienes, products of 5-lipoxygenase (5-LO) of AA, caused a specific stimulation of JNK. Together, our findings suggest that cPLA(2) mediates, at least partly, the signaling cascade by which Rac stimulates JNK.


Assuntos
Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosfolipases A/metabolismo , Proteínas rac de Ligação ao GTP/metabolismo , Animais , Araquidonato 5-Lipoxigenase/metabolismo , Ácido Araquidônico/farmacologia , Ácidos Araquidônicos/farmacologia , Sequência de Bases , Linhagem Celular , Citosol/enzimologia , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno , Leucotrienos/farmacologia , Oligodesoxirribonucleotídeos Antissenso/genética , Fosfolipases A/antagonistas & inibidores , Fosfolipases A/genética , Ratos , Transdução de Sinais , Transfecção
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