Capillary electrophoresis/mass spectrometry for the separation and characterization of bovine Cu,Zn‐superoxide dismutase

The native form of Cu,Zn‐superoxide dismutase (SOD‐1) is a homodimer that coordinates one Cu²⁺ and one Zn²⁺ per monomer. Cu²⁺ and Zn²⁺ ions play crucial roles in enzyme activity and structural stability, respectively. In addition, dimer formation is essential for SOD‐1 functionality, and in humans several SOD‐1 mutant isoforms have been associated with certain types of amyotrophic lateral sclerosis (ALS), a progressive neurodegenerative disorder. In this paper we used capillary electrophoresis and mass spectrometry to study the different structures of bovine SOD‐1. The metal ions of the native enzyme (Cu₂,Zn₂‐dimer SOD‐1) were released in acidic medium in order to obtain apo‐SOD‐1, which is a monomer. Both substances were analyzed by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) and capillary electrophoresis with ultraviolet and electrospray ionization mass spectrometry detection (CE/UV and CE/ESI‐MS, respectively). With MALDI‐TOF‐MS, using matrices of sinapinic acid (SA) or 2,5‐dihydroxybenzoic acid (DHB) with or without trifluoroacetic acid (TFA), similar mass spectra were obtained for the metalated and non‐metalated samples. In both cases, an average molecular mass corresponding to the apo‐monomer SOD‐1 was calculated. This finding indicated that the metals were released from the Cu₂,Zn₂‐dimer SOD‐1 during sample preparation or ionization. For CE/UV and CE/ESI‐MS, two background electrolytes (BGEs) potentially compatible with ESI‐MS detection were used, namely 1 M of acetic acid (pH 2.3) and 10 mM of ammonium acetate (pH 7.3). Using a sheath liquid of 2‐propanol/water (60:40 v/v), with or without 0.1% v/v of formic acid, CE/ESI‐MS sensitivity was enhanced when the acidic BGE and the acidic sheath liquid were used. However, the electrophoretic profiles and the mass spectra obtained suggested that the metals of Cu₂,Zn₂‐dimer SOD‐1 were released, which generated the apo‐monomer during the electrophoretic separation. The neutral BGE provided enhanced conditions for the detection of the native enzyme. The differences between the mass spectra obtained for the Cu₂,Zn₂‐dimer and the apo‐monomer forms were significant and the presence of formic acid in the sheath liquid affected only sensitivity. Our results highlight the importance of selecting appropriate non‐denaturing separation and detection conditions to obtain reliable structural information about non‐covalent protein complexes by CE/ESI‐MS. Copyright © 2010 John Wiley & Sons, Ltd.     

Searching for an extra neutral gauge boson from muon pair production at LHC

We search for signatures of the extra neutral gauge boson Z^′$Z^{\prime }$, predicted in some extensions of the Standard Model, from the analysis of some distributions for p+p→μ⁺+μ⁻+X$p+p\to {\mu }^{+}+{\mu }^{-}+X$, where the only exotic particle involved is Z^′$Z^{\prime }$. In addition to the invariant mass and charge asymmetry distributions, we propose in our search to use the transverse momentum distribution (pT$p_T$) as an observable. We do our calculation for two values of the LHC center of mass energy (7 and 14 TeV), corresponding to 1 and 100 fb⁻¹ of luminosity, in order to compare our findings from some models with the distributions following from the Standard Model. By applying convenient cuts in the invariant mass, we show that the final particles pT$p_T$ distributions can reveal the presence of an extra neutral gauge boson contribution. We also claim that it is possible to disentangle the models considered here and we emphasize that the minimal version of the model, based on SUC₍₃₎×SUL₍₃₎×UX₍₁₎$\mathit{SU}{(3)}_C\times \mathit{SU}{(3)}_L\times U{(1)}_X$ symmetry, presents the more clear signatures for Z^′$Z^{\prime }$ existence.     

Towards the control and inhibition of glycation-the role of the guanidine reaction center with aldehydic and diketonic dicarbonyls. A mass spectrometry study

Glycation of proteins by glucose and formation of end-stage adducts (AGEs, advanced glycation end products) has been implicated in pathological mechanisms associated with diabetic complications, macrovascular disease, chronic and renal insufficiency, Alzheimer's disease, and aging. Of the carbonyl containing compounds involved in this process, alpha-dicarbonyls have particular importance, being established as direct intermediates in the formation of well-known AGEs. The guanidino group, present in arginine residues, suffers direct modifications by sugars and its derivatives, and is considered to be an important chemical basis, targeting the control and inhibition of glycation.Seven dicarbonyl compounds, aldehydic and diketonic, were reacted with guanidine, in an attempt to establish structure/activity relationships. Electrospray mass spectrometry, together with tandem mass spectrometry, was used to identify and characterize the reaction products. The reactivity of guanidine was found to vary with the dicarbonyls used. For glyoxal, a high amount of dihydroxyimidazolidine was formed, whereas for methylglyoxal, dihydroxyimidazolidine was slowly converted into hydroimidazolone. Interestingly, aqueous guanidine was found to prevent argpyrimidine formation. The formation of several amine-dicarbonyl moieties was observed for the larger alkyl-diketonic dicarbonyls reaction systems, in particular. Molecular structures, bearing a polar chain, of an imidazole ring, and a nonpolar one, of alkyl groups, located at both sides of the imidazole rings, were attributed to these moieties. Gas-phase experiments suggested that the larger alkyl groups have a preference for being located at one of the sides of the imidazole rings. Moreover, the referred amine-dicarbonyl moieties are formed via (dihydroxyimidazolidine - 2H2O) moieties. The latter (dihydroxyimidazolidine - 2H2O) moieties are formed in high amounts in the larger alkyl-diketonic dicarbonyl reactions. Since these moieties react with dicarbonyl molecules, and react even faster with already modified amine functions, we can foresee that these species may be useful for controlling and inhibiting glycation of larger biomolecules, such as proteins.     

Enthalpy of formation of the cyclopentadienyl radical: photoacoustic calorimetry and ab initio studies

The gas-phase C-H bond dissociation enthalpy (BDE) in 1,3-cyclopentadiene has been determined by time-resolved photoacoustic calorimetry (TR-PAC) as 358 +/- 7 kJ mol(-1). Theoretical results from ab initio complete basis-set approaches, including the composite CBS-Q and CBS-QB3 procedures, and basis-set extrapolated coupled-cluster calculations (CCSD(T)) are reported. The CCSD(T) prediction for the C-H BDE of 1,3-cyclopentadiene (353.3 kJ mol(-1)) is in good agreement with the TR-PAC result. On the basis of the experimental and the theoretical values obtained, we recommend 355 +/- 8 kJ mol(-1) for the C-H BDE of 1,3-cyclopentadiene and 271 +/- 8 kJ mol(-1) for the enthalpy of formation of cyclopentadienyl radical.     

GKB-FP: an algorithm for large-scale discrete ill-posed problems

We describe an algorithm for large-scale discrete ill-posed problems, called GKB-FP, which combines the Golub-Kahan bidiagonalization algorithm with Tikhonov regularization in the generated Krylov subspace, with the regularization parameter for the projected problem being chosen by the fixed-point method by Bazán (Inverse Probl. 24(3), 2008). The fixed-point method selects as regularization parameter a fixed-point of the function ‖r _λ ‖₂/‖f _λ ‖₂, where f _λ is the regularized solution and r _λ is the corresponding residual. GKB-FP determines the sought fixed-point by computing a finite sequence of fixed-points of functions ||r_l^(k)||₂/||f_l^(k)||₂\|r_{\lambda}^{(k)}\|_{2}/\|f_{\lambda}^{(k)}\|_{2}, where f_l^(k)f_{\lambda}^{(k)} approximates f _λ in a k-dimensional Krylov subspace and r_l^(k)r_{\lambda}^{(k)} is the corresponding residual. Based on this and provided the sought fixed-point is reached, we prove that the regularized solutions f_l^(k)f_{\lambda}^{(k)} remain unchanged and therefore completely insensitive to the number of iterations. This and the performance of the method when applied to well-known test problems are illustrated numerically.     

Bounds for the number of points on curves over finite fields

Let U be a bounded open subset of ?^d, d ≥ 2 and f ∈ C(?U). The Dirichlet solution f^CU of the Dirichlet problem associated with the Laplace equation with a boundary condition f is not continuous on the closure ū of U in general if U is not regular but it is always Baire-one.Let H(U) be the space of all functions continuous on the closure ū and harmonic on U and F(H(U)) be the space of uniformly bounded absolutely convergent series of functions in H(U). We prove that f^CU can be obtained as a uniform limit of a sequence of functions in F(H(U)). Thus f^CU belongs to the subclass B_1/2 of Baire-one functions studied for example in [8]. This is not only an improvement of the result obtained in [10] but it also shows that the Dirichlet solution on the closure ū can share better properties than to be only a Baire-one function. Moreover, our proof is more elementary than that in [10].A generalization to the abstract context of simplicial function space on a metrizable compact space is provided.We conclude the paper with a brief discussion on the solvability of the abstract Dirichlet problem with a boundary condition belonging to the space of differences of bounded semicontinuous functions complementing the results obtained in [17].     

Use of 3D-transient analytical solution based on Green’s function to reduce computational time in inverse heat conduction problems

Inverse problems can be found in many areas of science and engineering and can be applied in different ways. Two examples can be cited: thermal properties estimation or heat flux function estimation in some engineering thermal process. Different techniques for the solution of inverse heat conduction problem (IHCP) can be found in literature. However, any inverse or optimization technique has a basic and common characteristic: the need to solve the direct problem solution several times. This characteristic is the cause of the great computational time consumed. In heat conduction problem, the time consumed is, usually, due to the use of numerical solutions of multidimensional models with refined mesh. In this case, if analytical solutions are available the computational time can be reduced drastically. This study presents the development and application of a 3D-transient analytical solution based on Green’s function. The inverse problem is due to the thermal properties estimation of conductors. The method is based on experimental determination of thermal conductivity and diffusivity using partially heated surface method without heat flux transducer. Originally developed to use numerical solution, this technique can, using analytical solution, estimate thermal properties faster and with better accuracy.     

$${{{\mathbb Z}_2}{{\mathbb Z}_4}}$$ -linear codes: generator matrices and duality

A code C{{\mathcal C}} is \mathbb Z₂\mathbb Z₄{{{\mathbb Z}_2}{{\mathbb Z}_4}} -additive if the set of coordinates can be partitioned into two subsets X and Y such that the punctured code of C{{\mathcal C}} by deleting the coordinates outside X (respectively, Y) is a binary linear code (respectively, a quaternary linear code). In this paper \mathbb Z₂\mathbb Z₄{{{\mathbb Z}_2}{{\mathbb Z}_4}} -additive codes are studied. Their corresponding binary images, via the Gray map, are \mathbb Z₂\mathbb Z₄{{{\mathbb Z}_2}{{\mathbb Z}_4}} -linear codes, which seem to be a very distinguished class of binary group codes. As for binary and quaternary linear codes, for these codes the fundamental parameters are found and standard forms for generator and parity-check matrices are given. In order to do this, the appropriate concept of duality for \mathbb Z₂\mathbb Z₄{{{\mathbb Z}_2}{{\mathbb Z}_4}} -additive codes is defined and the parameters of their dual codes are computed.     

Optimization of parameters of sampling and determination of reduced sulfur compounds using cryogenic capture and gas chromatography in tropical urban atmosphere

Reduced sulfur compounds, RSCs (H₂S, COS, CH₃SH, CH₃SCH₃, CS₂ and CH₃S₂CH₃) play a role in global cycle and acid rain formation. At trace levels RSCs in air are difficult to collect, store and analyze because of their highly adsorptive and reactive properties. This work optimizes parameters of sampling and instrumental determination of RSCs for urban measurements. The method used is based on cryogenic sampling and gas chromatography provided with a cryofocusing trap and flame photometric detection.Greater sampling efficiency was obtained with liquid argon as freezing fluid and air flow rate of 150 mL min^? 1 for two hours. Best results have been obtained with preconcentration for 3 min and injection volume of 3 ml. For H₂S, CH₃SH and CH₃S₂CH₃ the method showed a precision of 89%, limit of detection of 0.10 µg m^? 3 and limit of quantification 0.3 µg m^? 3. For CH₃SCH₃ and CS₂ the corresponding values were 89%, 0.15 µg m^? 3 and 0.5 µg m^? 3 and for COS were 75%, 0.18 µg m^? 3 and 0.8 µg m^? 3 respectively. Sampling efficiency varied between 70–80% for all the RSCs. Accuracy of H₂S from field measurements obtained with parallel measurements using a continuous monitor varied between 88 and 98%. The optimized methodology proved to be suitable for field measurements in urban tropical atmospheres with different characteristics.