RESUMEN
Reduction and oxidation reactions are essential for biochemical processes. They are part of metabolic pathways and signal transduction. Reactive oxygen species (ROS) as second messengers and oxidative modifications of cysteinyl (Cys) residues are key to transduce and translate intracellular and intercellular signals. Dysregulation of cellular redox signaling is known as oxidative distress, which has been linked to various pathologies, including neurodegeneration. Alzheimer's disease (AD) is a neurodegenerative pathology linked to both, abnormal amyloid precursor protein (APP) processing, generating Aß peptide, and Tau hyperphosphorylation and aggregation. Signs of oxidative distress in AD include: increase of ROS (H2O2, O2 â¢-), decrease of the levels or activities of antioxidant enzymes, abnormal oxidation of macromolecules related to elevated Aß production, and changes in mitochondrial homeostasis linked to Tau phosphorylation. Interestingly, Cys residues present in APP form disulfide bonds that are important for intermolecular interactions and might be involved in the aggregation of Aß. Moreover, two Cys residues in some Tau isoforms have been shown to be essential for Tau stabilization and its interaction with microtubules. Future research will show the complexities of Tau, its interactome, and the role that Cys residues play in the progression of AD. The specific modification of cysteinyl residues in redox signaling is also tightly connected to the regulation of various metabolic pathways. Many of these pathways have been found to be altered in AD, even at very early stages. In order to analyze the complex changes and underlying mechanisms, several AD models have been developed, including animal models, 2D and 3D cell culture, and ex-vivo studies of patient samples. The use of these models along with innovative, new redox analysis techniques are key to further understand the importance of the redox component in Alzheimer's disease and the identification of new therapeutic targets in the future.
RESUMEN
The dynamics and bifurcations of convective waves in rotating and buoyancy-driven spherical Rayleigh-Bénard convection are investigated numerically. The solution branches that arise as rotating waves (RWs) are traced by means of path-following methods, by varying the Rayleigh number as a control parameter for different rotation rates. The dependence of the azimuthal drift frequency of the RWs on the Ekman and Rayleigh numbers is determined and discussed. The influence of the rotation rate on the generation and stability of secondary branches is demonstrated. Multistability is typical in the parameter range considered.
RESUMEN
The multiplicity of stable convection patterns in a rotating spherical fluid shell heated from the inner boundary and driven by a central gravity field is presented. These solution branches that arise as rotating waves (RWs) are traced for varying Rayleigh number while their symmetry, stability, and bifurcations are studied. At increased Rayleigh numbers all the RWs undergo transitions to modulated rotating waves (MRWs) which are classified by their spatiotemporal symmetry. The generation of a third frequency for some of the MRWs is accompanied by a further loss of symmetry. Eventually a variety of MRWs, three-frequency solutions, and chaotic saddles and attractors control the dynamics for higher Rayleigh numbers.
Asunto(s)
Modelos Químicos , Reología/métodos , Soluciones/química , Simulación por ComputadorRESUMEN
Symmetry-breaking bifurcations have been studied for convection in a nonrotating spherical shell whose outer radius is twice the inner radius, under the influence of an externally applied central force field with a radial dependence proportional to 1/r(5). This work is motivated by the GeoFlow experiment, which is performed under microgravity condition at the International Space Station where this particular central force can be generated. In order to predict the observable patterns, simulations together with path-following techniques and stability computations have been applied. Branches of axisymmetric, octahedral, and seven-cell solutions have been traced. The bifurcations producing them have been identified and their stability ranges determined. At higher Rayleigh numbers, time-periodic states with a complex spatiotemporal symmetry are found, which we call breathing patterns.
RESUMEN
The linear stability of magnetohydrodynamic Taylor-Couette flows in axially unbounded cylinders is considered for magnetic Prandtl number unity. Magnetic background fields varying from purely axial to purely azimuthal are imposed, with a general helical field parametrized by ß=B(Ï)/B(z). We map out the transition from the standard magnetorotational instability (MRI) for ß=0 to the nonaxisymmetric azimuthal magnetorotational instability for ßâ∞. For finite ß, positive and negative wave numbers m , corresponding to right and left spirals, are no longer degenerate. For the nonaxisymmetric modes, the most unstable mode spirals in the opposite direction to the background field. The standard (ß=0) MRI is axisymmetric for weak fields (including the instability with the lowest Reynolds number) but is nonaxisymmetric for stronger fields. If the azimuthal field is due in part to an axial current flowing through the fluid itself (and not just along the central axis), then it is also unstable to the nonaxisymmetric Tayler instability which is most effective without rotation. For purely toroidal fields the solutions for m=±1 are identical so that in this case no preferred helicity results. For large ß the wave number m=-1 is preferred, whereas for ßâ²1 the mode with m=-2 is most unstable. The most unstable modes always spiral in the same direction as the background field. For background fields with positive and not too large ß the kinetic helicity of the fluctuations proves to be negative for all the magnetic instabilities considered.
RESUMEN
The clinical appearance of septic disorders is characterized by an enormous dynamic. The sepsis-induced dysbalance of the immune system necessitates immediate and aggressive therapeutic interventions to prevent further damage progression of the disease to septic shock and multiple organ failure. This includes supportive therapy to normalize and maintain organ and tissue perfusion as well as the identification of the infection focus. In cases where an infectious focus is identified, surgical source control frequently is a key element of the treatment strategy besides pharmacologic and supportive measures. The integrative approach of the management of septic patients requires rapid communication between the involved medical disciplines and the nursing personnel. Therefore, this article outlines current therapeutic concepts of septic diseases as well as central nursing aspects.
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Cuidados Críticos/métodos , Sepsis/cirugía , Acidosis/diagnóstico , Acidosis/terapia , Temperatura Corporal , Encefalopatías/diagnóstico , Encefalopatías/prevención & control , Cortisona/fisiología , Frecuencia Cardíaca , Humanos , Hidrocortisona/uso terapéutico , Hipotensión/diagnóstico , Hipoxia/diagnóstico , Control de Infecciones , Recuento de Leucocitos , Trastornos Mentales/etiología , Trastornos Mentales/terapia , Insuficiencia Multiorgánica/prevención & control , Complicaciones Posoperatorias/diagnóstico , Complicaciones Posoperatorias/prevención & control , Insuficiencia Renal/diagnóstico , Insuficiencia Renal/terapia , Sepsis/inmunología , Sepsis/enfermería , Sepsis/fisiopatología , Choque Séptico/inmunología , Choque Séptico/enfermería , Choque Séptico/fisiopatologíaRESUMEN
The Roberts flow, a helical flow in the form of convectionlike rolls, is known to be capable of both kinematic and nonlinear dynamo action. We study the Roberts dynamo with particular attention being paid to the spatial structure of the generated magnetic field and its back-reaction on the flow. The dynamo bifurcation is decisively determined by the symmetry group of the problem, which is given by a subgroup of discrete transformations and a continuous translational invariance of the flow. In the bifurcation the continuous symmetry is broken while the discrete subgroup symmetry completely survives. Its actions help in understanding the spatial structures of the magnetic field and of the modified flow. In accordance with experimental observations, the magnetic field component perpendicular to the originally invariant direction is much stronger than the component in this direction. Furthermore, the magnetic field is largely concentrated in layers separating the convectionlike rolls of the flow and containing, in particular, its stagnation points, which are isolated for the modified flow while they are line filling for the original Roberts flow. The magnetic field is strongest near beta-type stagnation points, with a two-dimensional unstable and a one-dimensional stable manifold, and is weak near alpha-type stagnation points, with a two-dimensional stable and a one-dimensional unstable manifold. This contrasts with the usual picture that dynamo action is promoted at the alpha points and impeded at the beta points. Both the creation of isolated stagnation points and the concentration of strong fields at the beta points may be understood as a result of the way in which the Roberts dynamo saturates. It is also found that, while the original Roberts flow is regular, the modified flow is chaotic in the layers between the convectionlike rolls where the magnetic field is concentrated. This chaoticity, which results from the back-reaction of the magnetic field on the flow, appears to merely enhance magnetic diffusion rather than to strengthen the dynamo effect.
RESUMEN
The Ku heterodimer is essential for the nonhomologous end-joining pathway of DNA double-strand break repair; it both protects the broken ends and recruits some of the many proteins required to complete repair. The recently determined structure of Ku provides insights into how it can both bind to the DNA ends and allow access by the other proteins required to rejoin them.
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Antígenos Nucleares , Daño del ADN , ADN Helicasas , Reparación del ADN , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/fisiología , Proteínas Nucleares/química , Proteínas Nucleares/fisiología , Animales , Rotura Cromosómica , Proteínas de Unión al ADN/genética , Humanos , Autoantígeno Ku , Proteínas Nucleares/genética , Estructura Cuaternaria de Proteína/genéticaRESUMEN
Rearrangement of gene segments to generate antigen receptor coding regions depends on the RAG1/2 recombinase, which assembles a synaptic complex between two DNA signal sequences and then cleaves the DNA directly adjacent to the paired signals. After coupled cleavage of complementary signal sequences, virtually all of the cleaved signal ends remained associated with RAG1/2 in stable complexes. These signal end complexes were distinct from various precleavage RAG1/2 signal complexes in that they were resistant to treatment with heparin. A mammalian joining apparatus consisting of purified Ku70/86, XRCC4, and DNA ligase IV proteins was sufficient to join deproteinized cleaved ends, but retention of signal sequences within the signal end complex blocked access to the DNA ends and prevented their joining by these proteins. Sequestration of cleaved ends within the signal end complex would account for the persistence of these ends in the cell after cleavage and may explain why they do not normally activate the DNA-damage-dependent cell cycle checkpoint.
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Antígenos Nucleares , ADN Helicasas , ADN Nucleotidiltransferasas/metabolismo , Proteínas de Unión al ADN/metabolismo , Proteínas de Homeodominio/metabolismo , Recombinación Genética , Secuencia de Bases , Cartilla de ADN , Hidrólisis , Autoantígeno Ku , Ligasas/metabolismo , Proteínas Nucleares/metabolismo , VDJ RecombinasasRESUMEN
The tumor suppressor Brca1 plays an important role in protecting mammalian cells against genomic instability, but little is known about its modes of action. In this work we demonstrate that recombinant human Brca1 protein binds strongly to DNA, an activity conferred by a domain in the center of the Brca1 polypeptide. As a result of this binding, Brca1 inhibits the nucleolytic activities of the Mre11/Rad50/Nbs1 complex, an enzyme implicated in numerous aspects of double-strand break repair. Brca1 displays a preference for branched DNA structures and forms protein-DNA complexes cooperatively between multiple DNA strands, but without DNA sequence specificity. This fundamental property of Brca1 may be an important part of its role in DNA repair and transcription.
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Proteína BRCA1/metabolismo , Proteínas de Ciclo Celular/metabolismo , Proteínas de Unión al ADN/metabolismo , ADN/metabolismo , Exodesoxirribonucleasas/metabolismo , Genes Supresores de Tumor , Proteínas Nucleares , Animales , Proteína BRCA1/fisiología , Sitios de Unión , Daño del ADN , Reparación del ADN , Humanos , Proteína Homóloga de MRE11 , Unión Proteica , Proteínas Recombinantes de Fusión/metabolismo , Proteínas Recombinantes de Fusión/fisiologíaRESUMEN
XRCC4 is essential for carrying out non-homologous DNA end joining (NHEJ) in all eukaryotes and, in particular, V(D)J recombination in vertebrates. Xrcc4 protein forms a complex with DNA ligase IV that rejoins two DNA ends in the last step of V(D)J recombination and NHEJ to repair double strand breaks. XRCC4-defective cells are extremely sensitive to ionizing radiation, and disruption of the XRCC4 gene results in embryonic lethality in mice. Here we report the crystal structure of a functional fragment of Xrcc4 at 2.7 A resolution. Xrcc4 protein forms a strikingly elongated dumb-bell-like tetramer. Each of the N-terminal globular head domains consists of a beta-sandwich and a potentially DNA-binding helix- turn-helix motif. The C-terminal stalk comprising a single alpha-helix >120 A in length is partly incorporated into a four-helix bundle in the Xrcc4 tetramer and partly involved in interacting with ligase IV. The Xrcc4 structure suggests a possible mode of coupling ligase IV association with DNA binding for effective ligation of DNA ends.
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Reparación del ADN , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/metabolismo , Secuencia de Aminoácidos , Animales , Proteínas Bacterianas/química , Sitios de Unión , Proteínas de Ciclo Celular/química , Cristalización , Cristalografía por Rayos X , ADN/química , ADN/metabolismo , ADN Ligasa (ATP) , ADN Ligasas/química , ADN Ligasas/metabolismo , Proteínas de Unión al ADN/genética , Dimerización , Humanos , Sustancias Macromoleculares , Ratones , Modelos Moleculares , Datos de Secuencia Molecular , Estructura Cuaternaria de Proteína , Saccharomyces cerevisiae/genética , Homología de Secuencia de AminoácidoRESUMEN
BACKGROUND: The response of eukaryotic cells to double-strand breaks in genomic DNA includes the sequestration of many factors into nuclear foci. Recently it has been reported that a member of the histone H2A family, H2AX, becomes extensively phosphorylated within 1-3 minutes of DNA damage and forms foci at break sites. RESULTS: In this work, we examine the role of H2AX phosphorylation in focus formation by several repair-related complexes, and investigate what factors may be involved in initiating this response. Using two different methods to create DNA double-strand breaks in human cells, we found that the repair factors Rad50 and Rad51 each colocalized with phosphorylated H2AX (gamma-H2AX) foci after DNA damage. The product of the tumor suppressor gene BRCA1 also colocalized with gamma-H2AX and was recruited to these sites before Rad50 or Rad51. Exposure of cells to the fungal inhibitor wortmannin eliminated focus formation by all repair factors examined, suggesting a role for the phosphoinositide (PI)-3 family of protein kinases in mediating this response. Wortmannin treatment was effective only when it was added early enough to prevent gamma-H2AX formation, indicating that gamma-H2AX is necessary for the recruitment of other factors to the sites of DNA damage. DNA repair-deficient cells exhibit a substantially reduced ability to increase the phosphorylation of H2AX in response to ionizing radiation, consistent with a role for gamma-H2AX in DNA repair. CONCLUSIONS: The pattern of gamma-H2AX foci that is established within a few minutes of DNA damage accounts for the patterns of Rad50, Rad51, and Brca1 foci seen much later during recovery from damage. The evidence presented strongly supports a role for the gamma-H2AX and the PI-3 protein kinase family in focus formation at sites of double-strand breaks and suggests the possibility of a change in chromatin structure accompanying double-strand break repair.
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Daño del ADN , Enzimas Reparadoras del ADN , Reparación del ADN , Histonas/metabolismo , Ácido Anhídrido Hidrolasas , Androstadienos/farmacología , Proteína BRCA1/genética , Proteína BRCA1/metabolismo , Bromodesoxiuridina/farmacología , Núcleo Celular/genética , Núcleo Celular/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Inhibidores Enzimáticos/farmacología , Rayos gamma , Humanos , Rayos Láser , Fosfatidilinositol 3-Quinasas/metabolismo , Inhibidores de las Quinasa Fosfoinosítidos-3 , Fosforilación , Recombinasa Rad51 , Células Tumorales Cultivadas , Rayos Ultravioleta , WortmaninaRESUMEN
During B and T cell development, the RAG1/RAG2 protein complex cleaves DNA at conserved recombination signal sequences (RSS) to initiate V(D)J recombination. RAG1/2 has also been shown to catalyze transpositional strand transfer of RSS-containing substrates into target DNA to form branched DNA intermediates. We show that RAG1/2 can resolve these intermediates by two pathways. RAG1/2 catalyzes hairpin formation on target DNA adjacent to transposed RSS ends in a manner consistent with a model leading to chromosome translocations. Alternatively, disintegration removes transposed donor DNA from the intermediate. At high magnesium concentrations, such as are present in mammalian cells, disintegration is the favored pathway of resolution. This may explain in part why RAG1/2-mediated transposition does not occur at high frequency in cells.
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Proteínas de Unión al ADN/genética , Genes RAG-1 , Proteínas de Homeodominio/genética , Recombinación Genética , Linfocitos B/fisiología , ADN/genética , Proteínas de Homeodominio/metabolismo , Humanos , Proteínas Nucleares , Linfocitos T/fisiología , Transposasas/genéticaRESUMEN
Repair of DNA double-strand breaks in vertebrate cells occurs mainly by an end-joining process that often generates junctions with sequence homologies of a few nucleotides. Mre11 is critical for this mode of repair in budding yeast and has been implicated in the microhomology-based joining. Here, we show that Mre11 exonuclease activity is sensitive to the presence of heterologous DNA, and to the structure and sequence of its ends. Addition of mismatched DNA ends stimulates degradation of DNA by Mre11, whereas cohesive ends strongly inhibit it. Furthermore, if a sequence identity is revealed during the course of degradation, it causes Mre11 nuclease activity to pause, thus stabilizing the junction at a site of microhomology. A nuclease-deficient Mre11 mutant that still binds DNA can also stimulate degradation by wild-type Mre11, suggesting that Mre11-DNA complexes may interact to bridge DNA ends and facilitate DNA joining.
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Reparación del ADN , Endodesoxirribonucleasas , Exodesoxirribonucleasas , Proteínas Fúngicas/fisiología , Proteínas de Saccharomyces cerevisiae , Catálisis , ADN/metabolismo , Exonucleasas/fisiologíaAsunto(s)
Antígenos Nucleares , ADN Helicasas , Reparación del ADN , Proteínas de Unión al ADN/fisiología , Proteínas Nucleares/fisiología , Envejecimiento/genética , Animales , Apoptosis , Diferenciación Celular , ADN , Reordenamiento Génico , Humanos , Autoantígeno Ku , Linfocitos/citología , Linfoma de Células B/genética , Ratones , Oncogenes , Proteína p53 Supresora de Tumor/genéticaRESUMEN
The Nijmegen breakage syndrome gene product (Nbs1) was shown recently to associate in vivo with the Mre11 and Rad50 proteins, which play pivotal roles in eukaryotic DNA double-strand break repair, meiotic recombination, and telomere maintenance. We show in this work that the triple complex of recombinant Nbs1, Mre11, and Rad50 proteins binds cooperatively to DNA and forms a distinct protein-DNA species. The Mre11/Rad50/Nbs1 complex displays several enzymatic activities that are not seen without Nbs1, including partial unwinding of a DNA duplex and efficient cleavage of fully paired hairpins. Unwinding and hairpin cleavage are both increased by the presence of ATP. On nonhairpin DNA ends, ATP controls a switch in endonuclease specificity that allows Mre11/Rad50/Nbs1 to cleave a 3'-protruding strand at a double-/single-strand transition. Mutational analysis demonstrates that Rad50 is responsible for ATP binding by the complex, but the ATP-dependent activities are expressed only with Nbs1 present.
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Adenosina Trifosfato/metabolismo , Antígenos Nucleares , Proteínas de Ciclo Celular/fisiología , ADN Helicasas/fisiología , Endodesoxirribonucleasas , Endonucleasas/metabolismo , Exodesoxirribonucleasas , Proteínas Fúngicas/fisiología , Adenosina Trifosfato/análogos & derivados , ADN/metabolismo , Reparación del ADN , ADN de Cadena Simple/metabolismo , Proteínas de Unión al ADN/metabolismo , Biblioteca de Genes , Células HeLa , Humanos , Autoantígeno Ku , Proteínas Nucleares/metabolismo , Proteínas Recombinantes de Fusión , Proteínas de Saccharomyces cerevisiae , Factores de TiempoRESUMEN
The F plasmid-carried bacterial toxin, the CcdB protein, is known to act on DNA gyrase in two different ways. CcdB poisons the gyrase-DNA complex, blocking the passage of polymerases and leading to double-strand breakage of the DNA. Alternatively, in cells that overexpress CcdB, the A subunit of DNA gyrase (GyrA) has been found as an inactive complex with CcdB. We have reconstituted the inactive GyrA-CcdB complex by denaturation and renaturation of the purified GyrA dimer in the presence of CcdB. This inactivating interaction involves the N-terminal domain of GyrA, because similar inactive complexes were formed by denaturing and renaturing N-terminal fragments of the GyrA protein in the presence of CcdB. Single amino acid mutations, both in GyrA and in CcdB, that prevent CcdB-induced DNA cleavage also prevent formation of the inactive complexes, indicating that some essential interaction sites of GyrA and of CcdB are common to both the poisoning and the inactivation processes. Whereas the lethal effect of CcdB is most probably due to poisoning of the gyrase-DNA complex, the inactivation pathway may prevent cell death through formation of a toxin-antitoxin-like complex between CcdB and newly translated GyrA subunits. Both poisoning and inactivation can be prevented and reversed in the presence of the F plasmid-encoded antidote, the CcdA protein. The products of treating the inactive GyrA-CcdB complex with CcdA are free GyrA and a CcdB-CcdA complex of approximately 44 kDa, which may correspond to a (CcdB)2(CcdA)2 heterotetramer.
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Proteínas Bacterianas/metabolismo , Toxinas Bacterianas/metabolismo , Citotoxinas/metabolismo , ADN-Topoisomerasas de Tipo II/metabolismo , ADN/metabolismo , Inhibidores de Topoisomerasa II , Proteínas Bacterianas/genética , Toxinas Bacterianas/genética , Girasa de ADN , ADN-Topoisomerasas de Tipo II/genética , Hidrólisis , Mutación , Unión ProteicaRESUMEN
Mammalian cells are protected from the effects of DNA double-strand breaks by end-joining repair. Cells lacking the Xrcc4 protein are hypersensitive to agents that induce DNA double-strand breaks, and are unable to complete V(D)J recombination. The residual repair of broken DNA ends in XRCC4-deficient cells requires short sequence homologies, thus possibly implicating Xrcc4 in end alignment. We show that Xrcc4 binds DNA, and prefers DNA with nicks or broken ends. Xrcc4 also binds to DNA ligase IV and enhances its joining activity. This stimulatory effect is shown to occur at the adenylation of the enzyme. DNA binding of Xrcc4 is correlated with its complementation of the V(D)J recombination defects in XRCC4-deficient cells, but is not required for stimulation of DNA ligase IV. Thus, the ability of Xrcc4 to bind to DNA suggests functions independent of DNA ligase IV.