RESUMO
Neural network quantum states (NQS) have been widely applied to spin-1/2 systems, where they have proven to be highly effective. The application to systems with larger on-site dimension, such as spin-1 or bosonic systems, has been explored less and predominantly using spin-1/2 Restricted Boltzmann Machines (RBMs) with a one-hot/unary encoding. Here, we propose a more direct generalization of RBMs for spin-1 that retains the key properties of the standard spin-1/2 RBM, specifically trivial product states representations, labeling freedom for the visible variables and gauge equivalence to the tensor network formulation. To test this new approach, we present variational Monte Carlo (VMC) calculations for the spin-1 anti-ferromagnetic Heisenberg (AFH) model and benchmark it against the one-hot/unary encoded RBM demonstrating that it achieves the same accuracy with substantially fewer variational parameters. Furthermore, we investigate how the hidden unit complexity of NQS depend on the local single-spin basis used. Exploiting the tensor network version of our RBM we construct an analytic NQS representation of the Affleck-Kennedy-Lieb-Tasaki (AKLT) state in the xyz spin-1 basis using only M=2N hidden units, compared to Mâ¼O(N2) required in the Sz basis. Additional VMC calculations provide strong evidence that the AKLT state in fact possesses an exact compact NQS representation in the xyz basis with only M=N hidden units. These insights help to further unravel how to most effectively adapt the NQS framework for more complex quantum systems.
RESUMO
We investigate how the presence of a single-particle mobility edge in a system can generate strong energy current rectification. Specifically, we study a quadratic bosonic chain subject to a quasiperiodic potential and coupled at its boundaries to spin baths of differing temperature. We find that rectification increases by orders of magnitude depending on the spatial position in the chain of localized eigenstates above the mobility edge. The largest enhancements occur when the coupling of one bath to the system is dominated by a localized eigenstate, while the other bath couples to numerous delocalized eigenstates. By tuning the parameters of the quasiperiodic potential it is thus possible to vary the amplitude, and even invert the direction, of the rectification.
RESUMO
Aminophospholipid (APL) trafficking across the plasma membrane is a key event in cell activation, apoptosis, and aging and is required for clearance of dying cells and coagulation. Currently the phospholipid molecular species externalized are unknown. Using a lipidomic method, we show that thrombin, collagen, or ionophore-activated human platelets externalize two phosphatidylserines (PSs) and five phosphatidylethanolamines (PEs). Four percent of the total cellular PE/PS pool (â¼300 ng/2 × 10(8) cells, thrombin), is externalized via calcium mobilization and protease-activated receptors-1 and -4, and 48% is contained in microparticles. Apoptosis and energy depletion (aging) externalized the same APLs in a calcium-dependent manner, and all stimuli externalized oxidized phospholipids, termed hydroxyeicosatetraenoic acid-PEs. Transmembrane protein-16F (TMEM-16F), the protein mutated in Scott syndrome, was required for PE/PS externalization during thrombin activation and energy depletion, but not apoptosis. Platelet-specific APLs optimally supported tissue factor-dependent coagulation in human plasma, vs. APL with longer or shorter fatty acyl chains. This finding demonstrates fatty acids as molecular determinants of APL that regulate hemostasis. Thus, the molecular species of externalized APL during platelet activation, apoptosis, and energy depletion were characterized, and their ability to support coagulation revealed. The findings have therapeutic implications for bleeding disorders and transfusion therapy. The assay could be applied to other cell events characterized by APL externalization, including cell division and vesiculation.
Assuntos
Coagulação Sanguínea , Plaquetas/metabolismo , Ácidos Graxos/química , Regulação da Expressão Gênica , Fosfolipídeos/química , Envelhecimento , Anexina A5/química , Apoptose , Biotinilação , Cálcio/metabolismo , Membrana Celular/metabolismo , Relação Dose-Resposta a Droga , Humanos , Trombina/química , Trombina/metabolismo , Fatores de TempoRESUMO
It has been known for many years that neutrophils and platelets participate in the pathogenesis of severe sepsis, but the inter-relationship between these players is completely unknown. We report several cellular events that led to enhanced trapping of bacteria in blood vessels: platelet TLR4 detected TLR4 ligands in blood and induced platelet binding to adherent neutrophils. This led to robust neutrophil activation and formation of neutrophil extracellular traps (NETs). Plasma from severely septic humans also induced TLR4-dependent platelet-neutrophil interactions, leading to the production of NETs. The NETs retained their integrity under flow conditions and ensnared bacteria within the vasculature. The entire event occurred primarily in the liver sinusoids and pulmonary capillaries, where NETs have the greatest capacity for bacterial trapping. We propose that platelet TLR4 is a threshold switch for this new bacterial trapping mechanism in severe sepsis.
Assuntos
Bactérias/imunologia , Plaquetas/imunologia , Neutrófilos/imunologia , Sepse/microbiologia , Sepse/fisiopatologia , Receptor 4 Toll-Like/metabolismo , Alanina Transaminase/sangue , Animais , Epitélio/patologia , Humanos , Lipopolissacarídeos/metabolismo , Fígado/metabolismo , Camundongos , Neutrófilos/enzimologia , Sepse/imunologiaRESUMO
Understanding the entropy production of systems strongly coupled to thermal baths is a core problem of both quantum thermodynamics and mesoscopic physics. While many techniques exist to accurately study entropy production in such systems, they typically require a microscopic description of the baths, which can become numerically intractable to study for large systems. Alternatively an open-systems approach can be employed with all the nuances associated with various levels of approximation. Recently, the mesoscopic leads approach has emerged as a powerful method for studying such quantum systems strongly coupled to multiple thermal baths. In this method, a set of discretized lead modes, each locally damped, provide a Markovian embedding. Here we show that this method proves extremely useful to describe entropy production of a strongly coupled open quantum system. We show numerically, for both noninteracting and interacting setups, that a system coupled to a single bath exhibits a thermal fixed point at the level of the embedding. This allows us to use various results from the thermodynamics of quantum dynamical semigroups to infer the nonequilibrium thermodynamics of the strongly coupled, non-Markovian central systems. In particular, we show that the entropy production in the transient regime recovers the well-established microscopic definitions of entropy production with a correction that can be computed explicitly for both the single- and multiple-lead cases.
RESUMO
Oxidized phospholipids (oxPLs) generated nonenzymatically display pleiotropic biological actions in inflammation. Their generation by cellular cyclooxygenases (COXs) is currently unknown. To determine whether platelets generate prostaglandin (PG)-containing oxPLs, then characterize their structures and mechanisms of formation, we applied precursor scanning-tandem mass spectrometry to lipid extracts of agonist-activated human platelets. Thrombin, collagen, or ionophore activation stimulated generation of families of PGs comprising PGE2 and D2 attached to four phosphatidylethanolamine (PE) phospholipids (16:0p/, 18:1p/, 18:0p/, and 18:0a/). They formed within 2 to 5 min of activation in a calcium, phospholipase C, p38 MAP kinases, MEK1, cPLA2, and src tyrosine kinase-dependent manner (28.1 ± 2.3 pg/2 × 108 platelets). Unlike free PGs, they remained cell associated, suggesting an autocrine mode of action. Their generation was inhibited by in vivo aspirin supplementation (75 mg/day) or in vitro COX-1 blockade. Inhibitors of fatty acyl reesterification blocked generation significantly, while purified COX-1 was unable to directly oxidize PE in vitro. This indicates that they form in platelets via rapid esterification of COX-1 derived PGE2/D2 into PE. In summary, COX-1 in human platelets acutely mediates membrane phospholipid oxidation via formation of PG-esterified PLs in response to pathophysiological agonists.
Assuntos
Aspirina/farmacologia , Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , Ciclo-Oxigenase 1/metabolismo , Inibidores de Ciclo-Oxigenase/farmacologia , Fosfolipídeos/metabolismo , Prostaglandinas/metabolismo , Plaquetas/fisiologia , Cálcio/metabolismo , Dinoprostona/metabolismo , Relação Dose-Resposta a Droga , Esterificação/efeitos dos fármacos , Retroalimentação Fisiológica/efeitos dos fármacos , Humanos , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , MAP Quinase Quinase 1/metabolismo , Fosfatidiletanolaminas/metabolismo , Ativação Plaquetária/efeitos dos fármacos , Prostaglandina D2/metabolismo , Proteína Quinase C/metabolismo , Receptor PAR-1/metabolismo , Trombina/metabolismo , Quinases da Família src/metabolismoRESUMO
5-Lipoxygenase (5-LOX) plays key roles in infection and allergic responses. Herein, four 5-LOX-derived lipids comprising 5-hydroxyeicosatetraenoic acid (HETE) attached to phospholipids (PLs), either phosphatidylethanolamine (PE) or phosphatidylcholine (18:0p/5-HETE-PE, 18:1p/5-HETE-PE, 16:0p/5-HETE-PE, and 16:0a/5-HETE-PC), were identified in primary human neutrophils. They formed within 2 minutes in response to serum-opsonized Staphylococcus epidermidis or f-methionine-leucine-phenylalanine, with priming by lipopolysaccharide, granulocyte macrophage colony-stimulating factor, or cytochalasin D. Levels generated were similar to free 5-HETE (0.37 ± 0.14 ng vs 0.55 ± 0.18 ng/10(6) cells, esterified vs free 5-HETE, respectively). They remained cell associated, localizing to nuclear and extranuclear membrane, and were formed by fast esterification of newly synthesized free 5-HETE. Generation also required Ca(2+), phospholipase C, cytosolic and secretory phospholipase A(2), 5-LOX activating protein, and mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1. 5-HETE-PLs were detected in murine S epidermidis peritonitis, paralleling neutrophil influx, and in effluent from Gram-positive human bacterial peritonitis. Formation of neutrophil extracellular traps was significantly enhanced by 5-LOX inhibition but attenuated by HETE-PE, whereas 5-HETE-PE enhanced superoxide and interleukin-8 generation. Thus, new molecular species of oxidized PL formed by human neutrophils during bacterial infection are identified and characterized.
Assuntos
Araquidonato 5-Lipoxigenase/metabolismo , Infecções Bacterianas/metabolismo , Eicosanoides/biossíntese , Neutrófilos/metabolismo , Idoso , Idoso de 80 Anos ou mais , Animais , Eicosanoides/química , Feminino , Infecções por Bactérias Gram-Positivas/metabolismo , Humanos , Ácidos Hidroxieicosatetraenoicos/biossíntese , Ácidos Hidroxieicosatetraenoicos/química , Técnicas In Vitro , Interleucina-8/biossíntese , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Neutrófilos/efeitos dos fármacos , Peritonite/metabolismo , Fosfolipídeos/biossíntese , Fosfolipídeos/química , Plasmalogênios/biossíntese , Plasmalogênios/química , Transdução de Sinais , Infecções Estafilocócicas/metabolismo , Staphylococcus epidermidis , Superóxidos/metabolismo , Espectrometria de Massas em Tandem , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
Deep insight can be gained into the nature of nonclassical correlations by studying the quantum operations that create them. Motivated by this we propose a measure of nonclassicality of a quantum operation utilizing the relative entropy to quantify its commutativity with the completely dephasing operation. We show that our measure of nonclassicality is a sum of two independent contributions, the generating power--its ability to produce nonclassical states out of classical ones, and the distinguishing power--its usefulness to a classical observer for distinguishing between classical and nonclassical states. Each of these effects can be exploited individually in quantum protocols. We further show that our measure leads to an interpretation of quantum discord as the difference in superdense coding capacities between a quantum state and the best classical state when both are produced at a source that makes a classical error during transmission.
RESUMO
BACKGROUND: Seasonal influenza A infection affects a significant cohort of the global population annually, resulting in considerable morbidity and mortality. Therapeutic strategies are of limited efficacy, and during a pandemic outbreak would only be available to a minority of the global population. Over-the-counter medicines are routinely taken by individuals suffering from influenza, but few studies have been conducted to determine their effectiveness in reducing pulmonary immunopathology or the influence they exert upon the generation of protective immunity. METHODS: A mouse model of influenza infection was utilised to assess the efficacy of paracetamol (acetaminophen) in reducing influenza-induced pathology and to examine whether paracetamol affects generation of protective immunity. RESULTS: Administration (intraperitoneal) of paracetamol significantly decreased the infiltration of inflammatory cells into the airway spaces, reduced pulmonary immunopathology associated with acute infection and improved the overall lung function of mice, without adversely affecting the induction of virus-specific adaptive responses. Mice treated with paracetamol exhibited an ability to resist a second infection with heterologous virus comparable with that of untreated mice. CONCLUSIONS: Our results demonstrate that paracetamol dramatically reduces the morbidity associated with influenza but does not compromise the development of adaptive immune responses. Overall, these data support the utility of paracetamol for reducing the clinical symptoms associated with influenza virus infection.
Assuntos
Acetaminofen/uso terapêutico , Anti-Inflamatórios não Esteroides/uso terapêutico , Vírus da Influenza A Subtipo H1N1 , Vírus da Influenza A Subtipo H3N2 , Infecções por Orthomyxoviridae/tratamento farmacológico , Infecções Respiratórias/tratamento farmacológico , Acetaminofen/farmacologia , Imunidade Adaptativa/efeitos dos fármacos , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Celecoxib , Inibidores de Ciclo-Oxigenase 2/uso terapêutico , Dinoprostona/metabolismo , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos/métodos , Feminino , Imunidade Inata/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/fisiopatologia , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/patologia , Infecções por Orthomyxoviridae/virologia , Pirazóis/uso terapêutico , Infecções Respiratórias/imunologia , Infecções Respiratórias/patologia , Infecções Respiratórias/virologia , Sulfonamidas/uso terapêutico , Carga Viral/efeitos dos fármacos , Eliminação de Partículas Virais/efeitos dos fármacosRESUMO
Activated platelets generate an eicosanoid proposed to be 8-hydroxy-9,10-dioxolane A3 (DXA3). Herein, we demonstrate that significant amounts of DXA3 are rapidly attached to phosphatidylethanolamine (PE) forming four esterified eicosanoids, 16:0p, 18:0p, 18:1p and 18:0a/DXA3-PEs that can activate neutrophil integrin expression. These lipids comprise the majority of DXA3 generated by platelets, are formed in ng amounts (24.3±6.1ng/2×108) and remain membrane bound. Pharmacological studies revealed DXA3-PE formation involves cyclooxygenase-1 (COX), protease-activated receptors (PAR) 1 and 4, cytosolic phospholipase A2 (cPLA2), phospholipase C and intracellular calcium. They are generated primarily via esterification of newly formed DXA3, but can also be formed in vitro via co-oxidation of PE during COX-1 co-oxidation of arachidonate. All four DXA3-PEs were detected in human clots. Purified platelet DXA3-PE activated neutrophil Mac-1 expression, independently of its hydrolysis to the free eicosanoid. This study demonstrates the structures and cellular synthetic pathway for a family of leukocyte-activating platelet phospholipids generated on acute activation, adding to the growing evidence that enzymatic PE oxidation is a physiological event in innate immune cells.
Assuntos
Plaquetas/metabolismo , Dioxolanos/sangue , Integrinas/sangue , Lipídeos/sangue , Fosfatidiletanolaminas/sangue , Cálcio/sangue , Ciclo-Oxigenase 1/sangue , Eicosanoides/sangue , Regulação da Expressão Gênica , Humanos , Integrinas/biossíntese , Antígeno de Macrófago 1/genética , Neutrófilos/metabolismo , Oxirredução , Fosfolipases A2 Citosólicas/sangue , Ativação Plaquetária/genética , Receptor PAR-1/sangue , Receptores de Trombina/sangue , Trombina/metabolismo , Fosfolipases Tipo C/sangueRESUMO
Nitration of unsaturated fatty acids such as linoleate by NO-derived reactive species forms novel derivatives (including nitrolinoleate [LNO2]) that can stimulate smooth muscle relaxation and block platelet activation by either NO/cGMP or cAMP-dependent mechanisms. Here, LNO2 was observed to inhibit human neutrophil function. LNO2, but not linoleic acid or the nitrated amino acid 3-nitrotyrosine, dose-dependently (0.2 to 1 micromol/L) inhibited superoxide (O2*-) generation, Ca2+ influx, elastase release, and CD11b expression in response to either phorbol 12-myristate 13-acetate or N-formyl-Met-Leu-Phe. LNO2 did not elevate cGMP, and inhibition of guanylate cyclase by 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one did not restore neutrophil responses, ruling out a role for NO. In contrast, LNO2 caused elevations in intracellular cAMP in the presence and absence of phosphodiesterase inhibition, suggesting activation of adenylate cyclase. Compared with phorbol 12-myristate 13-acetate-activated neutrophils, N-formyl-Met-Leu-Phe-activated neutrophils were more susceptible to the inhibitory effects of LNO2, indicating that LNO2 may inhibit signaling both upstream and downstream of protein kinase C. These data suggest novel signaling actions for LNO2 in mediating its potent inhibitory actions. Thus, nitration of lipids by NO-derived reactive species yields products with antiinflammatory properties, revealing a novel mechanism by which NO-derived nitrated biomolecules can influence the progression of vascular disease.
Assuntos
Degranulação Celular/efeitos dos fármacos , Integrinas/efeitos dos fármacos , Ácido Linoleico/farmacologia , Neutrófilos/efeitos dos fármacos , Nitrocompostos/farmacologia , Superóxidos/metabolismo , 1-Metil-3-Isobutilxantina/farmacologia , Anti-Inflamatórios/farmacologia , Cálcio/metabolismo , AMP Cíclico/metabolismo , GMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Humanos , Integrinas/biossíntese , Ácido Linoleico/química , N-Formilmetionina Leucil-Fenilalanina/farmacologia , NADPH Oxidases/metabolismo , Neutrófilos/metabolismo , Neutrófilos/fisiologia , Nitrocompostos/química , Inibidores de Fosfodiesterase/farmacologia , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
PGHS-2 (prostaglandin H synthase-2) is induced in mammalian cells by pro-inflammatory cytokines in tandem with iNOS [high-output ('inducible') nitric oxide synthase], and is co-localized with iNOS and nitrotyrosine in human atheroma macrophages. Herein, murine J774.2 macrophages incubated with lipopolysaccharide and interferon gamma showed induction of PGHS-2 and generated NO using iNOS that could be completely depleted by 12(S)-HPETE [12(S)-hydroperoxyeicosatetraenoic acid; 2.4 muM] or hydrogen peroxide (500 microM) (0.42+/-0.084 and 0.38+/-0.02 nmol x min(-1) x 10(6) cells(-1) for HPETE and H2O2 respectively). COS-7 cells transiently transfected with human PGHS-2 also showed HPETE- or H2O2-dependent NO decay (0.44+/-0.016 and 0.20+/-0.04 nmol x min(-1) x 10(6) cells(-1) for 2.4 microM HPETE and 500 microM H2O2 respectively). Finally, purified PGHS-2 consumed NO in the presence of HPETE or H2O2 (168 and 140 microM x min(-1) x microM enzyme(-1) for HPETE and H2O2 respectively), in a haem-dependent manner, with 20 nM enzyme consuming up to 4 microM NO. K(m) (app) values for NO and 15(S)-HPETE were 1.7+/-0.2 and 0.45+/-0.16 microM respectively. These data indicate that PGHS-2 catalytically consumes NO during peroxidase turnover and that pro-inflammatory cytokines simultaneously upregulate NO synthesis and degradation pathways in murine macrophages. Catalytic NO consumption by PGHS-2 represents a novel interaction between NO and PGHS-2 that may impact on the biological effects of NO in vascular signalling and inflammation.
Assuntos
Macrófagos/enzimologia , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico/deficiência , Óxido Nítrico/metabolismo , Peroxidase/metabolismo , Prostaglandina-Endoperóxido Sintases/metabolismo , Animais , Células COS , Catálise , Ciclo-Oxigenase 2 , Eletrodos , Indução Enzimática , Células HeLa , Humanos , Peróxido de Hidrogênio/farmacologia , Ácidos Hidroxieicosatetraenoicos/metabolismo , Inflamação/enzimologia , Inflamação/imunologia , Inflamação/metabolismo , Interferon gama/farmacologia , Cinética , Leucotrienos/metabolismo , Peróxidos Lipídicos/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Proteínas de Membrana , Camundongos , Óxido Nítrico Sintase Tipo II , Prostaglandina-Endoperóxido Sintases/genética , Prostaglandina-Endoperóxido Sintases/isolamento & purificação , TransfecçãoRESUMO
BACKGROUND AND PURPOSE: Airway microvascular leak (MVL) involves the extravasation of proteins from post-capillary venules into surrounding tissue. MVL is a cardinal sign of inflammation and an important feature of airway inflammatory diseases such as asthma. PGE2, a product of COX-mediated metabolism of arachidonic acid, binds to four receptors, termed EP14. PGE2 has a wide variety of effects within the airway, including modulation of inflammation, sensory nerve activation and airway tone. However, the effect of PGE2 on airway MVL and the receptor/s that mediate this have not been described. EXPERIMENTAL APPROACH: Evans Blue dye was used as a marker of airway MVL, and selective EP receptor agonists and antagonists were used alongside EP receptor-deficient mice to define the receptor subtype involved. KEY RESULTS: PGE2 induced significant airway MVL in mice and guinea pigs. A significant reduction in PGE2-induced MVL was demonstrated in Ptger2−/− and Ptger4−/− mice and in wild-type mice pretreated simultaneously with EP2 (PF-04418948) and EP4 (ER-819762) receptor antagonists. In a model of allergic asthma, an increase in airway levels of PGE2 was associated with a rise in MVL; this change was absent in Ptger2−/− and Ptger4−/− mice. CONCLUSIONS AND IMPLICATIONS: PGE2 is a key mediator produced by the lung and has widespread effects according to the EP receptor activated. Airway MVL represents a response to injury and under 'disease' conditions is a prominent feature of airway inflammation. The data presented highlight a key role for EP2 and EP4 receptors in MVL induced by PGE2.
Assuntos
Asma/metabolismo , Dinoprostona/metabolismo , Receptores de Prostaglandina E Subtipo EP2/metabolismo , Receptores de Prostaglandina E Subtipo EP4/metabolismo , Alérgenos , Animais , Azetidinas/farmacologia , Benzazepinas/farmacologia , Brônquios/metabolismo , Permeabilidade Capilar , Dinoprostona/análogos & derivados , Dinoprostona/farmacologia , Cobaias , Imidazóis/farmacologia , Masculino , Éteres Metílicos/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ovalbumina , Receptores de Prostaglandina E Subtipo EP2/agonistas , Receptores de Prostaglandina E Subtipo EP2/antagonistas & inibidores , Receptores de Prostaglandina E Subtipo EP2/genética , Receptores de Prostaglandina E Subtipo EP4/agonistas , Receptores de Prostaglandina E Subtipo EP4/antagonistas & inibidores , Receptores de Prostaglandina E Subtipo EP4/genética , Traqueia/metabolismoRESUMO
This protocol measures externalization of aminophospholipids (APLs) to the outside of the plasma membrane using mass spectrometry (MS). APL externalization occurs in numerous events, and it is relevant for transplant medicine, immunity and cancer. In this protocol, externalized APLs are chemically modified by using a cell-impermeable reagent (sulfo-NHS-biotin), and then they are isolated via a liquid:liquid extraction and quantified by reverse-phase liquid chromatography tandem MS (LC-MS/MS) against in-house-generated standards. This protocol describes a complementary method to existing assays that are not quantitative (e.g., annexin V flow cytometry), and it is applicable to the study of membrane reorganization in all cell types during apoptosis (e.g., during development, cancer, psychiatric disorders and other conditions, aging, vesiculation and cell division). The protocol takes â¼2-4 d, including the generation of standards.
Assuntos
Apoptose , Membrana Celular/metabolismo , Espectrometria de Massas/métodos , Ativação de Neutrófilo , Fosfatidiletanolaminas/análise , Fosfatidilserinas/análise , Ativação Plaquetária , Cromatografia Líquida de Alta Pressão , Humanos , Fosfatidiletanolaminas/metabolismo , Fosfatidilserinas/metabolismo , Espectrometria de Massas em TandemRESUMO
OBJECTIVE: To investigate the role of death receptor 3 (DR-3) and its ligand tumor necrosis factor-like molecule 1A (TL1A) in the early stages of inflammatory arthritis. METHODS: Antigen-induced arthritis (AIA) was generated in C57BL/6 mice deficient in the DR-3 gene (DR3(-/-) ) and their DR3(+/+) (wild-type) littermates by priming and intraarticular injection of methylated bovine serum albumin. The joints were sectioned and analyzed histochemically for damage to cartilage and expression of DR3, TL1A, Ly-6G (a marker for neutrophils), the gelatinase matrix metalloproteinase 9 (MMP-9), the aggrecanase ADAMTS-5, and the neutrophil chemoattractant CXCL1. In vitro production of MMP-9 was measured in cultures from fibroblasts, macrophages, and neutrophils following the addition of TL1A and other proinflammatory stimuli. RESULTS: DR3 expression was up-regulated in the joints of wild-type mice following generation of AIA. DR3(-/-) mice were protected against cartilage damage compared with wild-type mice, even at early time points prior to the main accumulation of Teff cells in the joint. Early protection against AIA in vivo correlated with reduced levels of MMP-9. In vitro, neutrophils were major producers of MMP-9, while neutrophil numbers were reduced in the joints of DR3(-/-) mice. However, TL1A neither induced MMP-9 release nor affected the survival of neutrophils. Instead, reduced levels of CXCL1 were observed in the joints of DR3(-/-) mice. CONCLUSION: DR-3 drives early cartilage destruction in the AIA model of inflammatory arthritis through the release of CXCL1, maximizing neutrophil recruitment to the joint and leading to enhanced local production of cartilage-destroying enzymes.
Assuntos
Artrite Experimental/metabolismo , Artrite Reumatoide/metabolismo , Cartilagem Articular/metabolismo , Membro 25 de Receptores de Fatores de Necrose Tumoral/metabolismo , Membro 15 da Superfamília de Ligantes de Fatores de Necrose Tumoral/metabolismo , Animais , Artrite Reumatoide/patologia , Cartilagem Articular/patologia , Quimiocina CXCL1/metabolismo , Fibroblastos/metabolismo , Humanos , Camundongos , Camundongos Knockout , Monócitos/metabolismo , Membrana Sinovial/metabolismoRESUMO
In some cases the state of a quantum system with a large number of subsystems can be approximated efficiently by the density-matrix renormalization group, which makes use of redundancies in the description of the state. Here we show that the achievable efficiency can be much better when performing density-matrix renormalization group calculations in the Heisenberg picture, as only the observable of interest but not the entire state is considered. In some nontrivial cases, this approach can even be exact for finite bond dimensions.
RESUMO
In this study, murine peritoneal macrophages from naïve lavage were found to generate four phospholipids that contain 12-hydroxyeicosatetraenoic acid (12-HETE). They comprise three plasmalogen and one diacyl phosphatidylethanolamines (PEs) (16:0p, 18:1p, 18:0p, and 18:0a at sn-1) and are absent in macrophages from 12/15-lipoxygenase (12/15-LOX)-deficient mice. They are generated acutely in response to calcium mobilization, are primarily cell-associated, and are detected on the outside of the plasma membrane. Levels of 12-HETE-PEs in naïve lavage are in a similar range to those of free 12-HETE (5.5 +/- 0.2 ng or 18.5 +/- 1.03 ng/lavage for esterified versus free, respectively). In healthy mice, 12/15-LOX-derived 12-HETE-PEs are found in the peritoneal cavity, peritoneal membrane, lymph node, and intestine, with a similar distribution to 12/15-LOX-derived 12-HETE. In vivo generation of 12-HETE-PEs occurs in a Th2-dependent model of murine lung inflammation associated with interleukin-4/interleukin-13 expression. In contrast, in Toll receptor-dependent peritonitis mediated either by live bacteria or bacterial products, 12-HETE-PEs are rapidly cleared during the acute phase then reappear during resolution. The human homolog, 18:0a/15-HETE-PE inhibited human monocyte generation of cytokines in response to lipopolysaccharide. In summary, a new family of lipid mediators generated by murine macrophages during Th2 inflammation are identified and structurally characterized. The studies suggest a new paradigm for lipids generated by 12/15-LOX in inflammation involving formation of esterified eicosanoids.
Assuntos
Eicosanoides/metabolismo , Ácidos Hidroxieicosatetraenoicos/química , Inflamação , Fosfatidiletanolaminas/metabolismo , Células Th2/metabolismo , Animais , Humanos , Lipídeos/química , Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Staphylococcus epidermidis/metabolismoRESUMO
To study the mechanisms involved in leukocyte recruitment induced by local bacterial infection within the CNS, we used intravital microscopy to visualize the interaction between leukocytes and the microvasculature in the brain. First, we showed that intracerebroventricular injection of LPS could cause significant rolling and adhesion of leukocytes in the brain postcapillary venules of wild-type mice, while negligible recruitment was observed in TLR4-deficient C57BL/10ScCr mice and CD14 knockout mice, suggesting recruitment is mediated by TLR4/CD14-bearing cells. Moreover, we observed reduced but not complete inhibition of recruitment in MyD88 knockout mice, indicating both MyD88-dependent and -independent pathways are involved. The leukocyte recruitment responses in chimeric mice with TLR4-positive microglia and endothelium, but TLR4-negative leukocytes, were comparable to normal wild-type mice, suggesting either endothelium or microglia play a crucial role in the induction of leukocyte recruitment. LPS injection induced both microglial and endothelial activation in the CNS. Furthermore, minocycline, an effective inhibitor of microglial activation, completely blocked the rolling and adhesion of leukocytes in the brain and blocked TNF-alpha production in response to LPS in vivo. Minocycline did not affect activation of endothelium by LPS in vitro. TNFR p55/p75 double knockout mice also exhibited significant reductions in both rolling and adhesion in response to LPS, indicating TNF-alpha signaling is critical for the leukocyte recruitment. Our results identify a TLR4 detection system within the blood-brain barrier. The microglia play the role of sentinel cells detecting LPS thereby inducing endothelial activation and leading to efficient leukocyte recruitment to the CNS.
Assuntos
Encéfalo/imunologia , Quimiotaxia de Leucócito/imunologia , Lipopolissacarídeos/imunologia , Microglia/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Barreira Hematoencefálica/imunologia , Encéfalo/irrigação sanguínea , Adesão Celular/imunologia , Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Imuno-Histoquímica , Receptores de Lipopolissacarídeos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microglia/imunologia , Fator 88 de Diferenciação Mieloide/deficiência , Receptor 4 Toll-Like/imunologia , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Localization of circulating lymphocytes to a site of inflammation is paramount for the development and maintenance of an immune response. In vitro studies using cell lines have previously demonstrated that rolling and adhesion of lymphocytes on endothelium requires CD44 interactions with hyaluronan (HA). To date, whether CD44 has a role in mediating CD4(+)-polarized T-helper 1 (Th1) and Th2 lymphocyte interactions with the endothelium in vivo is yet to be determined. In this study we used intravital microscopy to demonstrate that both Th1 and Th2 lymphocytes use CD44 to roll and adhere to tumor necrosis factor-alpha (TNFalpha)-activated microvasculature. Furthermore, chimeric studies imply that CD44 expression by both the endothelium and lymphocytes is essential for these interactions to occur. HA was also necessary for T cell-endothelial cell interactions in vivo and Th1 and Th2 cells rolled on immobilized HA in vitro via CD44. In vitro, both Th1 and Th2 lymphocytes have increased expression of CD44 and greater binding of fluorescent HA than naive cells. The interactions of Th1 and Th2 cells were entirely dependent upon both P-selectin and CD44 in vivo, but did not appear to be counter ligands in vitro. Taken together, these results suggest that CD44 and HA are key to both Th1 and Th2 lymphocyte interactions with the TNFalpha-activated endothelium and raises the possibility of cooperativity between the P-selectin/PSGL-1 and HA/CD44 pathways for Th1 and Th2 rolling in vivo.
Assuntos
Movimento Celular/imunologia , Endotélio Vascular/imunologia , Receptores de Hialuronatos/imunologia , Células Th1/imunologia , Células Th2/imunologia , Animais , Adesão Celular/imunologia , Comunicação Celular/imunologia , Endotélio Vascular/citologia , Regulação da Expressão Gênica/imunologia , Ácido Hialurônico/imunologia , Glicoproteínas de Membrana/imunologia , Camundongos , Microscopia de Vídeo , Selectina-P/imunologia , Células Th1/citologia , Células Th2/citologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
The detailed mechanisms by which acutely activated leukocytes metabolize NO and regulate its bioactivity are unknown. Therefore, healthy, chronic granulomatous disease (CGD) or myeloperoxidase (MPO)-deficient human neutrophils were examined for their ability to consume NO and attenuate its signaling. fMLP or PMA activation of healthy neutrophils caused NO consumption that was fully blocked by NADPH oxidase inhibition, and was absent in CGD neutrophils. Studies using MPO-deficient neutrophils, enzyme inhibitors, and reconstituted NADPH oxidase ruled out additional potential NO-consuming pathways, including Fenton chemistry, PGH synthase, lipoxygenase, or MPO. In particular, the inability of MPO to consume NO resulted from lack of H(2)O(2) substrate since all superoxide (O(2)(-.) reacted to form peroxynitrite. For healthy or MPO-deficient cells, NO consumption rates were 2- to 4-fold greater than O(2)(-.) generation, significantly faster than expected from 1:1 termination of NO with O(2)(-.). Finally, fMLP or PMA-stimulated NO consumption fully blocked NO-dependent neutrophil cGMP synthesis. These data reveal NADPH oxidase as the central regulator of NO signaling in human leukocytes. In addition, they demonstrate an important functional difference between CGD and either normal or MPO-deficient human neutrophils, namely their inability to metabolize NO which will alter their ability to adhere and migrate in vivo.