Highly Sensitive “Off/On” EPR Probes to Monitor Enzymatic Activity

The assessment of unregulated level of enzyme activity is a crucial parameter for early diagnoses in a wide range of pathologies. In this study, we propose the use of electron paramagnetic resonance (EPR) as an easy method to probe carboxylesterase (CE) enzymatic activity in vitro. For this application, were synthesized two amphiphilic, nitroxide containing esters, namely Tempo-C12 (T-C12) and Tempo-2-C12 (T-2-C12). They exhibit low solubility in water and form stable micelles in which the radicals are EPR almost silent, but the hydrolysis of the ester bond yields narrows and intense EPR signals. The intensity of the EPR signals is proportional to the enzymatic activity. CEs1, CEs2 and esterase from porcine liver (PLE) were investigated. The obtained results show that T-C12 and T-2-C12-containing systems display a much higher selectivity toward the CEs2, with a Limit of Detection of the same order of those ones obtained with optical methods.     

Partial Uniqueness and Obstruction to Uniqueness in Inverse Problems for Anisotropic Elastic Media

We consider the inverse problem of identifying the density and elastic moduli for three-dimensional anisotropic elastic bodies, given displacement and traction measurements made at their surface. These surface measurements are modelled by the dynamic Dirichlet-to-Neumann map on a finite time interval. For linear or nonlinear anisotropic hyperelastic bodies we show that the displacement-to-traction surface measurements do not change when the density and elasticity tensor in the interior are transformed tensorially by a change of coordinates fixing the surface of the body to first order. Our main tool, a new approach in inverse problems for elastic media, is the representation of the equations of motion in a covariant form (following Marsden and Hughes, 1983) that preserves the underlying physics.In the case of classical linear elastodynamics we then investigate how the type of anisotropy changes under coordinate transformations. That is, we analyze the orbits of general linear, anisotropic elasticity tensors under the action by pull-back of diffeomorphisms that fix the surface of the elastic body to first order, and derive a pointwise characterization of parts of the orbits under this action. For example, we show that the orbit of isotropic elastic media, at any point in the body, consists of some transversely isotropic and some orthotropic elastic media. We then derive the first uniqueness result in the inverse problem for anisotropic media using surface displacement-traction data: uniqueness of three elastic moduli for tensors in the orbit of isotropic elasticity tensors. ^†Partially supported by an MSRI Postdoctoral Fellowship. Research at MSRI is supported in part by NSF grant DMS-9850361. This work was conducted while the first author was a Gibbs Instructor at Yale University. ^‡Partially supported by an MSRI Postdoctoral Fellowship, and by NSF grant DMS-9801664 (9996350).     

Flame Induced Flow Features in the Presence of Darrieus-Landau Instability

The onset of hydrodynamic or Darrieus-Landau (DL) instability can largely impact on premixed flame morphology, turbulent flame speed and induced flow field. In this work, we focus on the latter induced flow by means of two dimensional direct numerical simulations (DNS) of slot burner flames performed in a parametric fashion. Results from linear stability analysis are used to select the adequate parameter range to be investigated. The presence of DL instability is initially assessed using a recently proposed statistical marker related to flame morphology. The differences between stable and unstable flames are then statistically investigated, utilizing a single, laminar, DL-induced corrugation as a reference state. Such DL-induced effects are investigated at various turbulence intensities, in terms of local propagation, induced strain rate patterns and flow field as well as vorticity production and transformation. Using displacement speed as a measure of local propagation, no noticeable statistical difference is observed between stable and unstable flames while strain rate and vorticity patterns are shown to be largely influenced by the DL induced morphology. From the modeling point view, an enhancement of counter gradient type transport for turbulent scalar fluxes is observed for hydrodynamically unstable flames.     

Mikrobestimmung von Siedepunkt und Schmelzpunkt, Mikrosublimation

Ohne Zusammenfassung     

Low molecular weight proteins in urines from healthy subjects as well as diabetic,nephropathic and diabetic‐nephropathic patients: a MALDI study

Urine samples from healthy subjects as well as diabetic, nephropathic and diabetic‐nephropathic patients were analyzed by matrix assisted laser desorption/ionization (MALDI) mass spectrometry in order to establish evidence of some possible differences in the peptide profile related to the pathological states. Multivariate analysis suggested the possibility of a distinction among the considered groups of patients. Some differences have been found, in particular, in the relative abundances of three ions at m/z 1912, 1219 and 2049. For these reasons, further investigation was carried out by MALDI/TOF/TOF to determine the sequence of these peptides and, consequently, to individuate their possible origin. By this approach, the peptide at m/z 1912 was found to originate from uromodulin, and its lower expression in the case of nephropathy can be well related to the pathological condition. Ions at m/z 2049 and 1219 originate from the collagen α‐1(I) chain precursor and from the collagen α‐5 (IV) chain precursor, respectively, and, also in this case, their different expressions can be related to the pathologies under investigation. The obtained data seem to indicate that urine is an interesting biological fluid to investigate on the peptide profile and to obtain, consequently, information on the dismetabolism activated by specific pathologies. Copyright © 2009 John Wiley & Sons, Ltd.     

Tuning Glutamine Binding Modes in Gd‐DOTA‐Based Probes for an Improved MRI Visualization of Tumor Cells

Three new magnetic resonance imaging probes that target glutamine transporters have been synthesized. They consist of a Gd‐DOTA‐monoamide moiety (DOTA=1,4,7,10‐tetraazacyclododecane‐1,4,7,10‐tetraacetic acid) linked through a six carbon atom chain to a vector represented by a glutamine residue bound through α‐carboxylic, γ‐carboxamidic, or α‐amino functionalities. Their uptake by HTC (rat hepatocarcinoma) and healthy rat hepatocytes has shown that the system containing the glutamine vector bound through the α‐carboxylic group displays a markedly higher affinity for tumor cells. The observed behavior is rationalized in terms of the exploitation of an additional glutamine transporter active in hepatic tumor cells.     

Phase behavior at low temperature of poly(tetrafluoroethylene) by temperature-modulated calorimetry

Temperature modulated differential calorimetry (TMDSC) is used to examine the crystal-crystal transitions of poly(tetrafluoroethylene). This study gives new information about the dynamic thermal behavior of such transitions. The involvement of reversible and irreversible processes during the phenomenon is observed, which are related to the order-disorder changes occurring during the transition.This study adds a new example to the response of TMDSC during first order transitions.     

Electrochemical and spectroscopic behavior of iron(III) porphyrazines in Langmuir-Schafer films

Thin films of a newly synthesized iron(III) porphyrazine, LFeOESPz ( L = ClEtO, OESPz = ethylsulfanylporphyrazine), have been deposited by the Langmuir-Schafer (LS) technique (horizontal lifting) on ITO or gold substrates. Before deposition, the floating films have been investigated at the air-water interface by pressure/area per molecule (pi/ A) experiments, Brewster angle microscopy (BAM) and UV-vis reflection spectroscopy (RefSpec). The complex reacts with water subphase (pH 6.2) forming the mu-oxo dimer, which becomes the predominant component of the LS films ( LS-Fe) as indicated by optical, IR, XPS, and electrochemical data. LS-Fe multilayers exhibit, between open circuit potential (OCP) and +0.90 V (vs SCE), two independent peak pairs with formal potentials, E surf (I) and E surf(II) of +0.56 V and +0.78 V, respectively. According to dynamic voltammetric and coulometric experiments the peak pair at +0.56 V is attributed to one-electron process at the iron(III) centers on the monomer, while the peak pair at +0.78 V is associated to a four-electron process involving mu-oxo-dimer oligomers. LS-Fe films prove to be quite stable electrochemically between OCP and +0.90 V. The electrochemical stability decreases, however, when the potential range is extended both anodically and cathodically outside these limits, due to formation of new species. Upon incubation with TCA solutions, LS-Fe films show remarkable changes in the UV-vis spectra, which are consistent with a significant mu-oxo dimer --> monomer conversion. Addition of TCA to the electrochemical cell using a LS-Fe film as working electrode, results in a linear increase of a cathodic current peak near -0.40 V as the TCA concentration varies in the 0.1-2.0 mM range. This behavior is interpreted in terms of TCA inducing a progressive change in the composition of the LS-Fe films in favor of the monomeric iron(III) porphyrazine, which is responsible for the observed increase in the cathodic current near -0.40 V.     

New insights from X-ray photoelectron spectroscopy into the chemistry of covalent enzyme immobilization, with glutamate dehydrogenase (GDH) on silicon dioxide as an example

A three-step process for immobilization of glutamate dehydrogenase (GDH) on the surface of silicon dioxide has been studied by X-ray photoelectron spectroscopy (XPS). The enzyme layer was deposited on the silicon dioxide surface after first exposing the surface to 3-aminopropyltriethoxysilane (3-APTS) and reacting the silylated surface with glutaraldehyde (GA). Fine XPS analysis, performed after each step of the chemical procedure, revealed unknown details of the step-by-step construction of the enzyme layer under different experimental conditions.