Studies on the formation of glutathionylcobalamin: any free intracellular aquacobalamin is likely to be rapidly and irreversibly converted to glutathionylcobalamin

A decade ago Jacobsen and co-workers reported the first evidence for the presence of glutathionylcobalamin (GSCbl) in mammalian cells and suggested that it could in fact be a precursor to the formation of the two coenzyme forms of vitamin B(12), adenosylcobalamin and methylcobalamin (Pezacka et al. Biochem. Biophys. Res. Commun. 1990, 169, 443). It has also recently been proposed by McCaddon and co-workers that GSCbl may be useful for the treatment of Alzheimer's disease (McCaddon et al. Neurology 2002, 58, 1395). Aquacobalamin is one of the major forms of vitamin B(12) isolated from mammalian cells, and high concentrations of glutathione (1-10 mM) are also found in cells. We have now determined observed equilibrium constants, K(obs)(GSCbl), for the formation of GSCbl from aquacobalamin and glutathione in the pH range 4.50-6.00. K(obs)(GSCbl) increases with increasing pH, and this increase is attributed to increasing amounts of the thiolate forms (RS(-)) of glutathione. An estimate for the equilibrium constant for the formation of GSCbl from aquacobalamin and the thiolate forms of glutathione of approximately 5 x 10(9) M(-1) is obtained from the data. Hence, under biological conditions the formation of GSCbl from aquacobalamin and glutathione is essentially irreversible. The rate of the reaction between aquacobalamin/hydroxycobalamin and glutathione for 4.50 < pH < 11.0 has also been studied and the observed rate constant for the reaction was found to decrease with increasing pH. The data were fitted to a mechanism in which each of the 3 macroscopic forms of glutathione present in this pH region react with aquacobalamin, giving k(1) = 18.5 M(-1) s(-1), k(2) = 28 +/- 10 M(-1) s(-1), and k(3) = 163 +/- 8 M(-1) s(-1). The temperature dependence of the observed rate constant at pH 7.40 ( approximately k(1)) was also studied, and activation parameters were obtained typical of a dissociative process (DeltaH++ = 81.0 +/- 0.5 kJ mol(-1) and DeltaS++ = 48 +/- 2 J K(-1) mol(-1)). Formation of GSCbl from aquacobalamin is rapid; for example, at approximately 5 mM concentrations of glutathione and at 37 degrees C, the half-life for formation of GSCbl from aquacobalamin and glutathione is 2.8 s. On the basis of our equilibrium and rate-constant data we conclude that, upon entering cells, any free (protein-unbound) aquacobalamin could be rapidly and irreversibly converted to GSCbl. GSCbl may indeed play an important role in vitamin B(12)-dependent processes.     

Existence globale pour une classe d'équations d'ondes perturbées

In this paper we prove a global well-posedness result for the following Cauchy problem:

where the initial data $\text{(f,g)∈}\text{H}\text{?}{}^1\text{(}\text{R}{}^3\text{)×L}{}^2\text{(}\text{R}{}^3\text{)}$ are compactly supported, 1?α<5, $\text{a}{}_{\mathrm{i}}\text{(t,x)∈L}{}^{\mathrm{\infty }}\text{(}\text{R}{}_{\mathrm{t}}\text{×}\text{R}{}_{\mathrm{x}}{}^3\text{)}$, $\text{V(t,x)∈L}{}^{\mathrm{\infty }}\text{(}\text{R}{}_{\mathrm{t}}\text{;L}{}^3\text{(}\text{R}{}^3{}_{\mathrm{x}}\text{))}$. To cite this article: N. Visciglia, C. R. Acad. Sci. Paris, Ser. I 338 (2004).     

-theory of -pseudo-differential algebras

We are concerned with the so-called -pseudo-differential calculus. We describe the spectrum of the unital and commutative -algebra given by the norm closure of the space of -order pseudo-differential operators modulo compact operators; other related algebras are also considered. Finally, their -theory is computed.

     

Degree complexity for a modified pigeonhole principle

 We consider a modification of the pigeonhole principle, M P H P, introduced by Goerdt in [7]. M P H P is defined over n pigeons and log n holes, and more than one pigeon can go into a hole (according to some rules). Using a technique of Razborov [9] and simplified by Impagliazzo, Pudlák and Sgall [8], we prove that any Polynomial Calculus refutation of a set of polynomials encoding the M P H P, requires degree Ω(log n). We also prove a simple Lemma giving a simulation of Resolution by Polynomial Calculus. Using this lemma, and a Resolution upper bound by Goerdt [7], we obtain that the degree lower bound is tight. Our lower bound establishes the optimality of the tree-like Resolution simulation by the Polynomial Calculus given in [6]. Received: 29 March 2001 / Published online: 2 September 2002 RID="⋆" ID="⋆" A prelimianry version appeared as part of the paper A Study of Proof Search Algorithms for Resolution and Polynomial Calculus published in the Proceedings of the 40-th IEEE Conference on Foundations of Computer Science, 1999 RID="†" ID="†" Partly supported by Project CICYT, TIC 98-0410-C02-01 and Promoción General del Conocimiento PB98-0937-C04-03. RID="††" ID="††" Part of this work was done while the author was a member of the School of Mathematics at Institute for Advanced Study supported by a NSF grant n. 9987845     

Large classes of minimally supported frequency wavelets of<Emphasis Type="Italic">L</Emphasis><Superscript>2</Superscript>(ℝ) and<Emphasis Type="Italic">H</Emphasis><Superscript>2</Superscript>(ℝ)

We introduce a new method to construct large classes of minimally supported frequency (MSF) wavelets of the Hardy space H ² (ℝ)and symmetric MSF wavelets of L ² (ℝ),and discuss the classification of such wavelets. As an application, we show that there are uncountably many such wavelet sets of L ² (ℝ)and H ² (ℝ).We also enumerate some of the symmetric wavelet sets of L ² (ℝ)and all wavelet sets of H ² (ℝ)consisting of three intervals. Finally, we construct families of MSF wavelets of L ₂ (ℝ)with Fourier transform even and not vanishing in any neighborhood of the origin.     

Digital holographic microscope with automatic focus tracking by detecting sample displacement in real time

We propose a new method for focus tracking during the recording of a sequence of digital holograms while the sample experiences axial displacement. Corrected reconstruction distances can be automatically calculated, and well-focused amplitude and phase-contrast images can be obtained for each digitized hologram. The method is demonstrated for inspection of microelectromechanical systems subjected to thermal load. The method can be applied as a quasi-real-time procedure.     

Understanding protease catalysed solid phase peptide synthesis

A protease (thermolysin) was used to directly synthesise a number of dipeptides from soluble Fmoc-amino acids onto a solid support (PEGA1900) in bulk aqueous media, often in very good yields. This shift in equilibrium toward synthesis is remarkable because for soluble dipeptides in aqueous solution hydrolysis rather than synthesis is observed. Three possible reasons for the equilibrium shift were considered: (i) using a solid support makes it easy to use an excess of reagents, so mass action contributes towards synthesis; (ii) reduction in the unfavourable hydrophobic hydration of the Fmoc group within the solid support compared with the free amino acid in solution and (iii) suppression of the ionization of amino groups linked to the solid phase due to mutual electrostatic repulsion. It was found that under the conditions studied the second effect was most important.     

Gauge-invariant temperature anisotropies and primordial non-Gaussianity

We provide the gauge-invariant expression for large-scale cosmic microwave background temperature fluctuations at second-order perturbation theory. This enables us to define unambiguously the nonlinearity parameter f(NL), which is used by experimental collaborations to pin down the level of non-Gaussianity in the temperature fluctuations. Furthermore, it contains a primordial term encoding all the information about the non-Gaussianity generated at primordial epochs and about the mechanism which gave rise to cosmological perturbations, thus neatly disentangling the primordial contribution to non-Gaussianity from the one caused by the postinflationary evolution.     

Theoretical study of a new DNA structure: the antiparallel Hoogsteen duplex

The structure of a new form of duplex DNA, the antiparallel Hoogsteen duplex, is studied in polyd(AT) sequences by means of state-of-the-art molecular dynamics simulations in aqueous solution. The structure, which was found to be stable in all of the simulations, has many similarities with the standard Watson-Crick duplex in terms of general structure, flexibility, and molecular recognition patterns. Accurate MM-PB/SA (and MM-GB/SA) analysis shows that the new structure has an effective energy similar to that of the B-type duplex, while it is slightly disfavored by intramolecular entropic considerations. Overall, MD simulations strongly suggest that the antiparallel Hoogsteen duplex is an accessible structure for a polyd(AT) sequence, which might compete under proper experimental conditions with normal B-DNA. MD simulations also suggest that chimeras containing Watson-Crick duplex and Hoogsteen antiparallel helices might coexist in a common structure, but with the differential characteristics of both type of structures preserved.