Computational Model of the Complex between GR113808 and the 5-HT4 Receptor Guided by Site-Directed Mutagenesis and the Crystal Structure of Rhodopsin

A computational model of the transmembrane domain of the human 5-HT₄ receptor complexed with the GR113808 antagonist was constructed from the crystal structure of rhodopsin and the putative residues of the ligand-binding site, experimentally determined by site-directed mutagenesis. The recognition mode of GR113808 consist of: (i) the ionic interaction between the protonated amine and Asp^3.32; (ii) the hydrogen bond between the carbonylic oxygen and Ser^5.43; (iii) the hydrogen bond between the ether oxygen and Asn^6.55; (iv) the hydrogen bond between the C-H groups adjacent to the protonated piperidine nitrogen and the electrons of Phe^6.51; and (v) the - aromatic-aromatic interaction between the indole ring and Phe^6.52.This computational model offers structural indications about the role of Asp^3.32, Ser^5.43, Phe^6.51, Phe^6.52, and Asn^6.55 in the experimental binding affinities. Asp^3.32Asn mutation does not affect the binding of GR113808 because the loss of binding affinity from an ion pair to a charged hydrogen bond is compensated by the larger energetical penalty of Asp to disrupt its side chain environment in the ligand-free form, and the larger interaction between Phe^6.51 and the piperidine ring of the ligand in the mutant receptor. In the Phe^6.52Val mutant the indole ring of the ligand replaces the interaction with Phe^6.52 by a similarly intense interaction with Tyr^5.38, with no significant effect in the binding of GR113808. The mutation of Asn^6.55 to Leu replaces the hydrogen bond of the ether oxygen of the ligand from Asn^6.55 to Cys^5.42, with a decrease of binding affinity that approximately equals the free energy difference between the SHO and NHO hydrogen bonds.Because these residues are also present in the other members of the neurotransmitter family of G protein-coupled receptors, these findings will also serve for our understanding of the binding of related ligands to their cognate receptors.     

Determination of perfluorocarboxylic acids in aqueous matrices by ion-pair solid-phase microextraction-in-port derivatization-gas chromatography-negative ion chemical ionization mass spectrometry

A rapid, selective and simple analytical procedure using tetrabutylammonium as ion pair in conjunction with solid-phase microextraction followed by in-port derivatization-GC-negative ion chemical ionization mass spectrometry was developed. The procedure allows an accurate determination of perfluoroalkylcarboxylic acids in aqueous samples at ng L(-1) levels (i.e. method detection limit 20 ng L(-1) forperfluorodecanoic acid) improving previous GC methods in terms of analysis time and sensitivity. Ammonia as reagent gas in the negative ion chemical ionization mass spectrometry increased the sensitivity at least 3-fold compared to methane for perfluorocarboxylic acid butyl esters. The developed procedure was successfully applied to effluents from wastewater treatment plants (i.e. 0.05-8.2 microg L(-1)) and harbor seawaters.     

Flow injection method for the fluorimetric determination of Zn with 8-(benzenesulphonamido) quinoline

The development of a fluorimetric method for the determination of zinc, based on the fluorescence of the zinc-8-(benzenesulphonamido) quinoline chelate in a micellar medium of sodium dodecylsulfate, is reported. The detection limit is 0.2 g l^–1, the working range is 0.5–700 g l^–1, and the sample throughput is 145 h^–1. The method was evaluated for the determination of zinc in food samples.A batch procedure for the simultaneous determination of zinc and cadmium, based on the synchronous and derivative spectra, is also proposed.     

Determination of aromatic hydrotropic drugs in pharmaceutical preparations by the surfactant-binding degree method

An aggregation parameter-based analytical approach, the surfactant-dye binding degree (SDBD) method, was used, for the first time, to determine aromatic hydrotropic compounds. The anionic dye Coomassie Brilliant Blue G (CBBG) was used as inductor of didodecyldimethylammonium bromide (DDABr) aggregates, whose formation was monitored from changes in the spectral features of the dye. Interactions between hydrotrope and DDABr molecules resulted in a decrease of the degree of binding of the cationic surfactant to CBBG, which was proportional to the concentration of hydrotrope in the aqueous solution. The CBBG-DDABr-hydrotrope chemical system was found to fit to the mathematical expression previously derived for the determination of amphiphilic compounds. The hydrotrope-surfactant bond strength determined the sensitivity achieved for the determination of hydrotropic compounds, which was highly dependent on the molecular structure of the analyte. The high precision (the relative standard deviation for 7 mg l(-1) of salicylic acid was 0.8%), rapidity (measurements were performed in a few minutes) and low cost (in both instrumentation and reactants) of the proposed method, made it especially suitable for quality control. The practical analytical applicability of the SDBD method for the control of hydrotropic drugs in pharmaceutical preparations was demonstrated by quantifying salicylic acid and acetyl salicylic acid in liquid (solutions) and solid (tablets, granulates, unguents, gels and creams) samples, which were directly analyzed after dissolution of the samples.     

The final steps of the Oppolzer cyclization: mechanism of the insertion of alkenes into allylpalladium(II) complexes

We report here the results of a computational study on the the mechanism of the Oppolzer cyclization. These results lead us to conclude that the insertion of olefins in Pd-allyl complexes probably takes place directly from the eta(3)-allyl species. The presence of a phosphane ligand in the reagents plays the role of enhancing the electron density on the Pd atom; this makes the alkene moiety more reactive towards insertion by back-donation from the metal. The results also indicate that the configuration of the new stereogenic centers is fixed in the insertion of the alkene into the (eta(3)-allyl)palladium species.     

Absolute electron-impact total ionization cross sections of chlorofluoromethanes

An experimental study is reported on the electron-impact total ionization cross sections (TICSs) of CCl4, CCl3F, CCl2F2, and CClF3 molecules. The kinetic energy of the colliding electrons was in the 10-85 eV range. TICSs were obtained as the sum of the partial ionization cross sections of all fragment ions, measured and identified in a linear double focusing time-of-flight mass spectrometer. The resulting TICS profiles--as a function of the electron-impact energy--have been compared both with those computed by ab initio and (semi)empirical methods and with the available experimental data. The computational methods used include the binary-encounter-Bethe (BEB) modified to include atoms with principal quantum numbers n> or =3, the Deutsch and M?rk (DM) formalism, and the modified additivity rule (MAR). It is concluded that both modified BEB and DM methods fit the experimental TICS for (CF4), CClF3, CCl2F2, CCl3F, and CCl4 to a high accuracy, in contrast with the poor accord of the MAR method. A discussion on the factors influencing the discrepancies of the fittings is presented.     

Chlorine K shell photoabsorption spectra of gas phase HCl and Cl2 molecules

High resolution photoabsorption spectra of HCl and Cl₂ have been measured near the chlorineK edge in the 2810–2850 eV photon energy range. Below the ClK edge, the strongest resonance is interpreted as a simple core excitation into the unoccupied σ* valence orbital for both molecules, leading to a markedly repulsive state. Higher resonances due to low lying Rydberg states, are observed in both systems, but with a larger oscillator strength for HCl as compared to Cl₂. In Cl₂, the σ* orbital is deep enough to avoid any mixing with Rydberg orbitals. In HCl, we observe the dipole forbidden Cl 1s → 4s transition which denotes a strong 4s–4p hybridization. Above the ClK edge, the multiplet features seen for HCl are analysed in terms of double-core-valence excited vacancy states. In Cl₂, their counterpart are found very close to the ionization threshold because of the deep σ* orbital and possibly because the excited core and valence electrons originates either from the same atomic site or from different ones.     

The Role of Aqueous Aerosols in the “Glyoxylate Scenario”: An Experimental Approach

The origin of life is one of the fundamental questions in science. Eschenmoser proposed the “glyoxylate scenario”, in which plausible abiotic synthesis pathways were suggested to be compatible with the constraints of prebiotic chemistry. In this proposal, the stem compound is HCN. In this work, we explore the “glyoxylate scenario” through several syntheses of HCN polymers, paying particular attention to the role of the aqueous aerosols, together with statistical methods, as a step to elucidate the synthetic problem of the origin of life. The soluble and insoluble HCN polymers synthetized were analyzed by GC‐MS. We identified, for the first time, glyoxylic acid in these polymers, together with some constituents of the reductive tricarboxylic acid cycle, amino acids and several N‐heterocycles. The findings presented herein, as the first global approach to the “glyoxylate scenario”, give full effect to this hypothesis and prove that aqueous aerosols could play an important role in this plausible scene of the origin of life.