Circular dichroism of the photoreceptor pigment oxyblepharismin

Circular dichroism (CD) was used to study the structure of oxyblepharismin (OxyBP), the photoreceptor chromophore for the photophobic response of the blue form of Blepharisma japonicum. Both the chromophore associated to its native protein and the free chromophore in ethanol solution were investigated. CD spectra in the far-UV range indicate that OxyBP induces a slight increase in the alpha-helix content of the protein matrix. CD spectra in the near-UV and visible region of the spectrum show that OxyBP adopts a chiral conformation with a preferential geometry not only when associated to its protein matrix, but also when isolated and dissolved in ethanol. This experimental result is related to the existence of a high-energy interconversion barrier between two enantiomeric structures of the molecule and discussed on the basis of an asymmetric biosynthesis of its precursor, blepharismin.     

Difference between Extra‐ and Intracellular T1 Values of Carboxylic Acids Affects the Quantitative Analysis of Cellular Kinetics by Hyperpolarized NMR

Incomplete knowledge of the longitudinal relaxation time constant (T₁) leads to incorrect assumptions in quantitative kinetic models of cellular systems, studied by hyperpolarized real‐time NMR. Using an assay that measures the intracellular signal of small carboxylic acids in living cells, the intracellular T₁ of the carboxylic acid moiety of acetate, keto‐isocaproate, pyruvate, and butyrate was determined. The intracellular T₁ is shown to be up to four‐fold shorter than the extracellular T₁. Such a large difference in T₁ values between the inside and the outside of the cell has significant influence on the quantification of intracellular metabolic activity. It is expected that the significantly shorter T₁ value of the carboxylic moieties inside cells is a result of macromolecular crowding. An artificial cytosol has been prepared and applied to predict the T₁ of other carboxylic acids. We demonstrate the value of this prediction tool.     

Efficient Hyperpolarization of U‐13C‐Glucose Using Narrow‐Line UV‐Generated Labile Free Radicals

Free radicals generated by UV‐light irradiation of a frozen solution containing a fraction of pyruvic acid (PA) have demonstrated their dissolution dynamic nuclear polarization (dDNP) potential, providing up to 30 % [1‐¹³C]PA liquid‐state polarization. Moreover, their labile nature has proven to pave a way to nuclear polarization storage and transport. Herein, differently from the case of PA, the issue of providing dDNP UV‐radical precursors (trimethylpyruvic acid and its methyl‐deuterated form) not involved in any metabolic pathway was investigated. The ¹³C dDNP performance was evaluated for hyperpolarization of [U‐¹³C₆,1,2,3,4,5,6,6‐d₇]‐d ‐glucose. The generated UV‐radicals proved to be versatile and highly efficient polarizing agents, providing, after dissolution and transfer (10 s), a ¹³C liquid‐state polarization of up to 32 %.     

Oxidative rearrangement of indoles: a new approach to the EFHG-tetracyclic core of diazonamide A

A new approach to the ring EFHG-tetracyclic core fragment of the marine secondary metabolite diazonamide A is described. The route is based on the oxidative rearrangement of 3-arylindole-2-carboxylates. Thus, a range of 3-arylindole-2-carboxylates (3, 8) underwent rearrangement to the corresponding 3,3-disubstituted oxindoles (4, 9) with migration of the ester group upon treatment with tert-butyl hypochlorite followed by acid. The oxindoles 9 with a 3-[2-(4-methoxybenzyloxy)]phenyl substituent underwent cyclization to the tetracyclic aminals 11 following N-protection, reduction, and treatment with methanesulfonic anhydride. The methodology was applied to the tyrosine-indole derivative 17 to give the EFHG-tetracyclic core of diazonamide A.     

Visible Multiphoton Dissociation of Chromophore-Tagged Peptides

The visible photodissociation mechanisms of QSY7-tagged peptides of increasing size have been investigated by coupling a mass spectrometer and an optical parametric oscillator laser beam. The experiments herein consist of energy resolved collision- and laser-induced dissociation measurements on the chromophore-tagged peptides. The results show that fragmentation occurs by similar channels in both activation methods, but that the branching ratios are vastly different. Observation of a size-dependent minimum laser pulse energy required to induce fragmentation, and collisional cooling rates in time resolved experiments show that laser-induced dissociation occurs through the absorption of multiple photons by the chromophore and the subsequent heating through vibrational energy redistribution. The differences in branching ratio between collision- and laser-induced dissociation can then be understood by the highly anisotropic energy distribution following absorption of a photon.

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The effective medium and the average field approximations vis-à-vis the Hashin-Shtrikman bounds. II. The generalized self-consistent scheme in matrix-based composites

The present Part II of this two-part study is concerned with the average field approximation (AFA), and the effective medium approximation (EMA) in two-phase matrix-based dielectric composites through the use of an auxiliary configuration in which a particle of the inclusion phase is first surrounded by some matrix material, and then embedded in the effective medium. Those models will be referred as the generalized self-consistent scheme-average field approximation (GSCS-AFA), and the generalized self-consistent scheme-effective medium approximation (GSCS-EMA). We show that there are four types of the GSCS-AFA and a single type of the GSCS-EMA. In this paper the application of those models to dielectric composites with isotropic constituents and an inclusion phase that consists of randomly oriented ellipsoidal particles will be studied. The analytical solution of the auxiliary problem, which consists of an ellipsoidal particle confocally surrounded by a matrix shell and embedded in the effective medium, is achieved by means of ellipsoidal harmonics. Our results show that the effective property predictions of the GSCS-EMA and GSCS-AFA for the considered systems differ from each other, and more importantly, out of the four GSCS-AFA models, three of them violate the Hashin-Shtrikman bounds. The predictions of the GSCS-EMA obey the bounds. It is then shown that the version of the GSCS-AFA which obeys the Hashin-Shtrikman bounds for an inclusion phase with randomly oriented ellipsoids will violate them in the case of a particle shape which is not simply connected. Moreover, it turns out that the SCS-AFA studied in Part I also violates the Hashin-Shtrikman bounds in that case; the EMA, as expected, owing to its realizability property, continues to obey the bounds. Among the AFA and EMA in matrix-based composites, the GSCS-EMA therefore stands out as the method to be recommended.     

On two models of arbitrarily curved three-dimensional thin interphases in elasticity

In boundary value problems involving thin interphases, it is often desirable to have a model of an interphase which makes possible to solve for the fields in the adjacent media without having to solve for the fields in it. This is usually achieved in the literature by replacing the interphase by a geometrical surface with appropriately designed “imperfect interface” conditions on it. In the present study, carried out in the setting of elasticity, another option is explored: the geometry of the interphase is left intact, and conditions are devised for the displacements and tractions pertaining to the media adjacent to the interphase and evaluated at both sides of it such that they will simulate the presence of the interphase. Those conditions do not involve the fields within the interphase, yet they depend on its material properties and on those of the adjacent media as well, and make possible to solve for the fields in the adjacent media without having to solve for the fields in the interphase. The formulation is given in a parallel orthogonal curvilinear coordinate system suitable for the modeling arbitrarily curved three-dimensional interphases of constant thickness. Both types of the above described interphase models are tested in the setting of a coated infinite fiber embedded in a matrix which is subjected to an anti-plane shear loading and an in-plane transverse shear loading at infinity, and their predictions are compared with the exact solutions for the fields in the three-phase configuration consisting of the interphase and its adjacent media. The model in which the interphase geometry is left intact is observed to perform generally better than the one in which the interphase is replaced by an interface.     

Mechanistic Insights about the Ligand-Enabled Oxy-arylation/vinylation of Alkenes via Au(I)/Au(III) Catalysis

The mechanism of oxy-arylation/vinylation of alkenes catalyzed by the (MeDalphos)AuCl complex was comprehensively investigated by DFT. (P,N)Au(Ph)²⁺ and (P,N)Au(vinyl)²⁺ are key intermediates accounting for the activation of the alkenols and for their cyclization by outer-sphere nucleophilic attack of oxygen. The 5-exo and 6-endo paths have been computed and compared, reproducing the peculiar regioselectivity difference observed experimentally between 4-penten-1-ol, (E) and (Z)-4-hexen-1-ols. Examining the way the alkenol coordinates to gold (more η² or η¹) can offer, in some cases, a simple way to predict the favored path of cyclization.     

Hydrogen Peroxide-Responsive Triggers Based on Borinic Acids: Molecular Insights into the Control of Oxidative Rearrangement

Arylborinic acids represent new, efficient, and underexplored hydrogen peroxide-responsive triggers. In contrast to boronic acids, two concomitant oxidative rearrangements are involved in the complete oxidation of these species, which might represent a major limitation for an efficient effector (drug or fluorophore) release. Herein, a comprehensive study of H₂O₂-mediated unsymmetrical arylborinic acid oxidation to investigate the factors that could selectively guide their oxidative rearrangement is described. The o-CF₃ substituent was found to be an excellent directing group allowing a complete regioselectivity on borinic acid models. This result was successfully applied to synthesizing new borinic acid-based fluorogenic probes, which exclusively release the fluorescent moiety upon H₂O₂ treatment. These compounds maintained their superior kinetic properties compared to boronic acids, thus further enhancing the potential of arylborinic acids as valuable new H₂O₂-sensitive triggers.     

Marinobufagenin extraction from Rhinella marina toad glands: Alternative approaches for a systematized strategy

Marinobufagenin is a bufadienolide compound detected mainly in skin and parotoid gland secretions of Rhinella marina (L.) toad. Bufadienolides regulate the Na⁺/K⁺‐ATPase pump by inhibiting the cardiotonic steroid dependent‐site and act as cardiac inotropes with vasoconstrictive properties. Marinobufagenin and other bufadienolides, such as telocinobufagin and bufalin, are thought to be found endogenously in mammals in salt‐sensitive hypertensive states such as essential hypertension, congestive heart‐failure, and preeclampsia. The role of marinobufagenin as antimicrobial agent and its cytotoxic potential have also been recognized. The particular interest around marinobufagenin prompts us to consider the Rhinella marina toad venom as a possible source for molecules with pharmacological and/or diagnostic potential. In this article, two different approaches of extraction and purification of marinobufagenin from Rhinella marina (L.) venom are studied: (i) Preparative thin‐layer chromatography combined to mass spectrometry and/or ultraviolet detection and (ii) solid‐phase extraction coupled with fractionation on high‐performance liquid chromatography. Different chromatographic conditions are tested for each approach. The solid‐phase extraction combined with high‐performance liquid chromatography fractionation approach was preferred as it offered a greater yield, was less time‐consuming and allowed us to selectively isolate marinobufagenin. Both protocols aim to provide efficient and convenient methods for toad venom extraction, based on an easily automatable and systematized strategy.