RESUMEN
Configuration transitions of individual molecules and atoms on surfaces are traditionally described using an Arrhenius equation with energy barrier and pre-exponential factor (attempt rate) parameters. Characteristic parameters can vary even for identical systems, and pre-exponential factors sometimes differ by orders of magnitude. Using low-temperature scanning tunnelling microscopy (STM) to measure an individual dibutyl sulfide molecule on Au(111), we show that the differences arise when the relative position of tip apex and molecule changes by a fraction of the molecule size. Altering the tip position on that scale modifies the transition's barrier and attempt rate in a highly correlated fashion, which results in a single-molecular enthalpy-entropy compensation. Conversely, appropriately positioning the STM tip allows selecting the operating point on the compensation line and modifying the transition rates. The results highlight the need to consider entropy in transition rates of single molecules, even at low temperatures.
RESUMEN
For a fixed 2 µm×2 µm area of a Co/Pt-CoO perpendicular exchange bias system we image the ferromagnetic (FM) domains for various applied fields with 10-nm resolution by magnetic force microscopy (MFM). Using quantitative MFM we measure the local areal density of pinned uncompensated spins (pinUCS) in the antiferromagnetic (AFM) CoO layer and correlate the FM domain structure with the UCS density. Larger applied fields drive the receding domains to areas of proportionally higher pinUCS aligned antiparallel to FM moments. The data confirm that the evolution of the FM domains is determined by the pinUCS in the AFM layer, and also present examples of frustration in the system.
RESUMEN
We have developed an optical cantilever deflection detector with a spot size <3 µm and fm Hz(-1/2) sensitivity over a>10 MHz bandwidth. In this work, we demonstrate its potential for detecting small-amplitude oscillations of various flexural and torsional oscillation modes of cantilevers. The high deflection sensitivity of the interferometer is particularly useful for detecting cantilever oscillations in aqueous solutions, enabling us to reach the thermal noise limit in scanning or atomic force microscopy experiments with stiff cantilevers. This has resulted in atomic-resolution images of solid-liquid interfaces and submolecular-resolution images of native membranes.
RESUMEN
The resonance frequency and the excitation amplitude of a silicon cantilever have been measured as a function of distance to a cleaved KBr(001) surface with a low-temperature scanning force microscope (SFM) in ultrahigh vacuum. We identify two regimes of tip-sample distances. Above a site-dependent critical tip-sample distance reproducible data with low noise and no interaction-induced energy dissipation are measured. In this regime reproducible SFM images can be recorded. At closer tip-sample distances, above two distinct atomic sites, the frequency values jump between two limiting curves on a timescale of tens of milliseconds. Furthermore, additional energy dissipation occurs wherever jumps are observed. We attribute both phenomena to rarely occurring changes in the tip apex configuration which are affected by short-range interactions with the sample. Their respective magnitudes are related to each other. A specific candidate two-level system is also proposed.
RESUMEN
200-nm-thick Ni films in an epitaxial Cu/Ni/Cu/Si(001) structure are expected to have an in-plane effective magnetic anisotropy. However, the in-plane remanence is only 42%, and magnetic force microscopy domain images suggest perpendicular magnetization. Quantitative magnetic force microscopy analysis can resolve the inconsistencies and show that (i) the films have perpendicular domains capped by closure domains with magnetization canted at 51 degrees from the film normal, (ii) the magnetization in the Bloch domain walls between the perpendicular domains accounts for the low in-plane remanence, and (iii) the perpendicular magnetization process requires a short-range domain wall motion prior to wall-magnetization rotation and is nonhysteretic, whereas the in-plane magnetization requires long-range motion before domain-magnetization rotation and is hysteretic.
RESUMEN
Atomic Force Microscopy (AFM) was used to investigate the ultrastructural appearance of transverse wood cell wall surfaces in embedded and polished Norway spruce wood blocks. The prepared surfaces showed only little height differences, suitable for high resolution AFM phase contrast imaging. Our results revealed randomly arranged wood cell wall components in the thick secondary 2 (S2) layers of the tracheid cell walls. It is concluded that the observed distribution pattern of the cellulose fibril/matrix structure is close to the original cell wall structure. In this context, the plasticity of wood cell wall components to re-arrange and adjust to different conditions resulting in diverse structural pattern is discussed.
Asunto(s)
Pared Celular/ultraestructura , Microscopía de Fuerza Atómica/métodos , Madera/ultraestructura , Microtomía/métodos , Modelos Biológicos , Picea/ultraestructura , Manejo de Especímenes/métodos , Propiedades de Superficie , Adhesión del Tejido/métodosAsunto(s)
Apoptosis , Caspasas/metabolismo , Necrosis , Transducción de Señal , Apoptosis/genética , Apoptosis/fisiología , Muerte Celular/genética , Activación Enzimática , Genes Transgénicos Suicidas , Genes Virales , Humanos , Mutación , Necrosis/patología , Necrosis/fisiopatología , Neoplasias/patología , Investigación , Transducción de Señal/fisiologíaRESUMEN
We propose and apply to the KBr(001) surface a new procedure for species recognition in scanning force microscopy (SFM) of ionic crystal surfaces which show a high symmetry of the charge arrangement. The method is based on a comparison between atomistic simulations and site-specific frequency versus distance measurements. First, by taking the difference of force-distance curves extracted at a few judiciously chosen surface sites we eliminate site-independent long-range forces. The obtained short-range force differences are then compared with calculated ones assuming plausible tip apex models. This procedure allows for the first time identification of the tip apex polarity and of the positive and negative sublattices in SFM images of the (001) cleavage surface of an ionic crystal with the rock salt structure.
RESUMEN
We have developed a sensitive method for the detection of recombinant antibody-antigen interactions in a microarray format. The biochip sensor platform used in this study is based on an oriented streptavidin monolayer that provides a biological interface with well-defined surface architecture that dramatically reduces nonspecific binding interactions. All the antibody or antigen probes were biotinylated and coupled onto streptavidin-coated biochip surfaces (1 microL total volume). The detection limits for the immobilized probes on the microarray surface were 0.5 microgram/mL (200 fmol/spot) for the peptide antigen and 0.1 microgram/mL (3 fmol/spot) for the recombinant antibodies. Optimal concentrations for the detection of the Cy5-labeled protein target were in the range of 20 micrograms/mL. Protein microchips were used to measure antibody-antigen kinetics, to find optimal temperature conditions, and to establish the shelf life of recombinant antibodies immobilized on the streptavidin surface. For recombinant antibody fragments with a kDa of 10-100 nM, we have established an easy and direct immunoassay. In addition, we developed an indirect method for antibody detection with no need for expensive and time-consuming antibody purifications and modifications. Such a method was shown to be useful for large-scale screening of recombinant antibody fragments directly after their functional expression in bacteria. Our data demonstrate that recombinant antibody fragments are suitable components in the construction of antibody chips.
Asunto(s)
Anticuerpos/química , Antígenos/análisis , Antígenos/química , Análisis por Matrices de Proteínas/métodos , Estreptavidina/química , Complejo Antígeno-Anticuerpo/análisis , Complejo Antígeno-Anticuerpo/química , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Biotinilación/métodos , Carbocianinas , Materiales Biocompatibles Revestidos , Escherichia coli/química , Escherichia coli/metabolismo , Oro , Inmunoensayo/métodos , Fragmentos de Inmunoglobulinas/química , Sondas Moleculares/química , Análisis por Matrices de Proteínas/instrumentación , Proteínas Recombinantes de Fusión/análisis , Proteínas Recombinantes de Fusión/química , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Magnetic force microscopy (MFM) measurements were performed on an exchange-biased CoO/(CoPt) multilayer sample at 7.5 K. Applying an external magnetic field of up to 7 T saturates the ferromagnetic layer and the remaining uncompensated antiferromagnetic spins at the antiferromagnet-ferromagnet interfaces are imaged with high lateral resolution. The coupling between the uncompensated spins and the spins in the ferromagnet are found to be antiferromagnetic. Quantitative analysis of the MFM images revealed that 7% of the spins at the interface are uncompensated and contribute to the exchange biasing.
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BACKGROUND: HIPK2 (homeodomain-interacting protein kinase 2) has been identified as a nuclear serine/threonine kinase. A central function of HIPK2 is repressing transcription of homeodomain containing transcription factors. RESULTS AND CONCLUSIONS: We show here that HIPK2 activates transcription mediated by tumor suppressor p53 responsive promoter elements. Overexpression of HIPK2 leads to an increase of p53 protein expression or stability, which becomes enhanced further in the presence of the DNA damaging drug doxorubicin. The effects of HIPK2 on p53 are not observed with kinase deficient HIPK2 mutants. However, HIPK2 is not sufficient for phosphorylation of three crucial serine residues of p53, suggesting that HIPK2-induced p53 activation does not involve phosphorylation of p53. Instead, HIPK2 leads to a downregulation of p53-induced Mdm2 protein and this may lead to stabilization of p53. Overexpression of HIPK2 does not lead to a change of Mdm2 mRNA expression. The data suggest that HIPK2 plays a critical role in p53 mediated cellular responses by removing the p53 inhibitor protein Mdm2 via modification of the protein itself or its intracellular movement.
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We report direct force measurements of the formation of a chemical bond. The experiments were performed using a low-temperature atomic force microscope, a silicon tip, and a silicon (111) 7x7 surface. The measured site-dependent attractive short-range force, which attains a maximum value of 2.1 nanonewtons, is in good agreement with first-principles calculations of an incipient covalent bond in an analogous model system. The resolution was sufficient to distinguish differences in the interaction potential between inequivalent adatoms, demonstrating the ability of atomic force microscopy to provide quantitative, atomic-scale information on surface chemical reactivity.
RESUMEN
MOTIVATION: We devise a computational model using protein-protein interactions. RESULTS: Peptide-antibody interactions can be used to perform a large number of small logical operations in parallel. We show for example how a sequence of operations can be used to compare the number of occurrences of an element in two sets and how to estimate the number of occurrences of an element in a set. Similar to DNA-computing, these techniques could in principle be extended to solve instances of NP-complete problems. We give as an example a procedure to solve examples of the satisfiability problem.
Asunto(s)
Computadores , Péptidos/química , Proteínas/metabolismo , Anticuerpos/inmunología , Reacciones Antígeno-Anticuerpo , Simulación por Computador , Modelos Moleculares , Péptidos/inmunología , Unión Proteica , Proteínas/inmunologíaRESUMEN
HIPK2 (homeodomain-interacting protein kinase 2) is a CD95 binding partner in yeast. Its primary amino acid sequence is highly conserved between human and mouse. The highest HIPK2 mRNA expression is found in neuronal tissue. The HIPK2 gene is located on human chromosome 7q33-35 and the protein is mainly localized in the nucleus. HIPK2 has been described to play a role as a co-repressor for homeodomain transcription factors.
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Proteínas Portadoras/genética , Cromosomas Humanos Par 7 , Proteínas Nucleares/genética , Proteínas Serina-Treonina Quinasas/genética , Receptor fas/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteínas Portadoras/metabolismo , Codón Iniciador , ADN Complementario , Femenino , Expresión Génica , Humanos , Masculino , Ratones , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , ARN Mensajero , Homología de Secuencia de Aminoácido , Distribución Tisular , Células Tumorales CultivadasRESUMEN
HIPK2 has been described as a homoedomain-interacting protein kinase with a nuclear localization. Here we describe that HIPK2 can also associate with TRADD, a protein that interacts with tumor necrosis factor receptor type 1 (TNF-R1). Under the conditions where HIPK2/TRADD association was found, no direct interaction of HIPK2 with CD95, TNF-R1, FADD or caspase-8 could be detected. Therefore, HIPK2 may play a role in TNF-R1 mediated signaling.
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Proteínas Portadoras/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas , Proteínas/metabolismo , Receptores del Factor de Necrosis Tumoral/metabolismo , Receptor fas/metabolismo , Anticuerpos Monoclonales/metabolismo , Western Blotting , Caspasa 8 , Caspasa 9 , Caspasas/metabolismo , Línea Celular , Núcleo Celular/metabolismo , ADN Complementario/metabolismo , Proteína Ligando Fas , Humanos , Plásmidos/metabolismo , Pruebas de Precipitina , Unión Proteica , Transducción de Señal , Factor 1 Asociado a Receptor de TNF , TransfecciónRESUMEN
Standard orthodontic facebows may accidentally detach from the appliance buccal tubes at night; this could reduce the effectiveness of extra oral traction and occasionally cause an injury. To try and prevent facebow detachment at night a facebow with a locking mechanism was introduced. This study assessed the ability of 706 consecutively treated patients to learn to wear and use this facebow. The facebows were fitted in 9 different practices supervised by 12 orthodontists. Data from the patients and orthodontists were collected over a 2-year period and covered approximately 166,550 nights. All the orthodontists were able to fit and adjust the facebow; a total of 697 patients successfully used the facebow. Accidental detachment of the facebow at night was reported to be less than 1%. This indicates a significant improvement in the safety of the facebow and should help to improve compliance by increasing the number of hours of wear achieved by the patients.
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Aparatos de Tracción Extraoral , Diseño de Aparato Ortodóncico , Prevención de Accidentes , Adolescente , Adulto , Niño , Falla de Equipo , Estudios de Evaluación como Asunto , Aparatos de Tracción Extraoral/efectos adversos , Aparatos de Tracción Extraoral/clasificación , Femenino , Estudios de Seguimiento , Humanos , Masculino , Aparatos Ortodóncicos Funcionales , Aparatos Ortodóncicos Removibles , Cooperación del Paciente , Seguridad , Propiedades de SuperficieRESUMEN
Caspases (cysteine aspartate-specific proteases) are a structurally related group of cysteine proteases that cleave peptide bonds following specific recognition sequences. They play a central role in activating apoptosis of vertebrate cells. To measure apoptosis induced by various stimuli and at an early apoptotic stage, caspases are an ideal target. This is especially the case when apoptotic cells have to be analyzed ex vivo before phagocytes remove them. A new and sensitive caspase assay is based on a substrate that contains only aspartate residues linked to rhodamine 110. With this and similar substrates, we are able to detect intracellular caspase activation by flow cytometry after apoptosis induction in intact hematopoetic cell lines.
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Aminoácidos/metabolismo , Apoptosis , Caspasas/metabolismo , Colorantes Fluorescentes/metabolismo , Células Jurkat/citología , Células Jurkat/enzimología , Péptidos/metabolismo , Rodaminas/metabolismo , Clorometilcetonas de Aminoácidos/metabolismo , Anticuerpos Monoclonales/farmacología , Apoptosis/inmunología , Caspasa 3 , Inhibidores de Caspasas , Inhibidores de Cisteína Proteinasa/metabolismo , Activación Enzimática , Humanos , Hidrólisis , Células Jurkat/metabolismo , Poli(ADP-Ribosa) Polimerasas/metabolismo , Espectrometría de Fluorescencia , Estaurosporina/farmacología , Especificidad por Sustrato , Receptor fas/inmunologíaRESUMEN
Wilson's disease can result in fulminant liver failure due to hepatic copper overload. The CD95 system mediates apoptosis and has been demonstrated to be involved in liver disease. In this study CD95 mediated apoptosis was investigated in patients with fulminant hepatic failure in the course of Wilson's disease and in an in vitro model of copper treated human hepatoma cells. In patients, hepatic expression of CD95 and CD95L mRNA and apoptosis were detected. Copper overload in vitro resulted in hepatocytic apoptosis which could be reduced with a neutralizing anti-CD95L antibody. Copper treatment of hepatocytes results in activation of the CD95 system and induction of apoptosis which is operative during the course of hepatic failure in acute Wilson's disease.
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Apoptosis , Cobre/toxicidad , Encefalopatía Hepática/etiología , Degeneración Hepatolenticular/fisiopatología , Receptor fas/metabolismo , Enfermedad Aguda , Proteína Ligando Fas , Expresión Génica , Humanos , Hígado/patología , Glicoproteínas de Membrana/metabolismo , Proteína p53 Supresora de TumorRESUMEN
Oxidative stress has been associated with the induction of programmed cell death. The CD95 ligand/receptor system is a specific mediator of apoptosis. We have used the model of drug-induced apoptosis to assess whether the CD95 ligand mRNA is induced by reactive oxygen intermediates. Treatment of HepG2 hepatoma cells with bleomycin induced the production of reactive oxygen intermediates and, as an additional parameter of oxidative stress, resulted in glutathione (GSH) depletion. In parallel, CD95 ligand mRNA expression was induced. In a similar fashion CD95 ligand mRNA expression increased after treatment with H2O2. Additional treatment with the antioxidant and GSH precursor N-acetylcysteine resulted in partial restoration of intracellular GSH levels and in reduced induction of CD95 ligand mRNA. Induction of CD95 ligand mRNA by bleomycin was further reduced by combined treatment with N-acetylcysteine and deferoxamine. These data suggest a direct role of oxygen radicals in the induction of the CD95 ligand.