Preliminary studies on the surface layer of starch

Characterization of the surface layer of starch powder, originated from potato or maize, was carried out applying various experimental techniques: Fourier-transformed infrared internal reflection spectroscopy (FTIR-IRS), inverse gas chromatography (IGC) and scanning electron microscopy (SEM). The main kinds of starch were studied as potential fillers for rubber and results discussed in terms of factors requiring modification to improve starch miscibility and activity towards rubber. Starch of high amylopectine content shows surface enrichment with amylose what makes crystallinity of the surface layer higher than in bulk of the material. Particles of the both kinds of starch have very smooth surface (morphological index, I_m = 0.995), highly resistant to its geometrical development. Dispersive component of the surface energy of starch was found to be similar to that of low energy polymers (γ_s^d = 32.5 mJ/m² at 23°C) and its surface to be of electron-donor character.     

Acetogenic isoquinoline alkaloids: CXII. Separation and identification of dimeric naphthylisoquinoline alkaloids by liquid chromatography coupled to electrospray ionization mass spectrometry

The atropodiastereomeric dimeric naphthylisoquinoline alkaloids, michellamines A (1a), B (1b) and C (1c), together with their monomers, korupensamines A (2a) and B (2b), were investigated using electrospray ionization tandem mass spectrometry coupled to liquid chromatography (LC–ESI-MS–MS). From the spectra obtained, characteristic product ions were chosen to monitor the chromatographic separation achieved on an RP-18 column. Under acidic conditions required for chromatographic analysis, the monomeric alkaloids 2a and 2b yielded protonated molecules [M+H]⁺, while the dimers, the michellamines, exhibited doubly protonated [M+2H]²⁺ molecules. In addition, the coeluting alkaloids 1b and 2b were identified unambiguously by means of tandem mass spectrometry. Thus, together with the retention times of the alkaloids, the product ion spectra allowed us the identification of michellamines in the presence of their presumed biogenetic monomeric precursors. Application of the HPLC–MS–MS method successfully proved the enzymatic formation of michellamine C (1c) by in vitro dimerization of korupensamine B (2b).     

A sensitive and selective determination method of histamine by HPLC with intramolecular excimer-forming derivatization and fluorescence detection

A highly sensitive, selective and simple method is described for the determination of histamine by high-performance liquid chromatography (HPLC) with fluorescence detection. The method is based on an intramolecular excimer-forming fluorescence derivatization of histamine with 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), followed by reversed-phase HPLC. Histamine, having two amino moieties in a molecule, was converted to the dipyrene-labeled derivative by reaction with PSE. The derivative afforded intramolecular excimer fluorescence (450-540 nm), which can clearly be discriminated from the monomer fluorescence (370-420 nm) emitted from PSE. Typically, a 10 micro L sample solution was mixed with 100 micro L of derivatization reagent solution, which was a mixture of 0.5 mm PSE in acetonitrile and 0.5 mm potassium carbonate in water (8:2, v/v). The derivatization was carried out at 100 degrees C for 90 min. The PSE derivative of histamine could be separated by reversed-phase ODS column with isocratic elution using acetonitrile:water (82:18, v/v) containing 0.03% triethylamine. The detection limit (singnal-to-noise ratio = 3) of histamine was 0.5 fmol for a 30 micro L injection. The method was successfully applied to the determination of histamine in human urine, and had enough selectivity and sensitivity for urinary histamine quantification.     

Monosaccharide templates for de novo designed 4-alpha-helix bundle proteins: template effects in carboproteins

De novo design and total chemical synthesis of proteins provide powerful approaches to critically test our understanding of protein folding, structure, and stability. The 4-alpha-helix bundle is a frequently studied structure in which four amphiphilic alpha-helical peptide strands form a hydrophobic core. Assembly of protein models on a template has been suggested as a way to reduce the entropy of folding. We have previously developed the concept of carbohydrates as templates in the de novo design of protein models termed 'carboproteins'. Here we present the chemical synthesis of three 8.1 kDa 4-alpha-helix bundles by oxime ligation of tetra-aminooxyacetyl functionalized D-galacto-, D-gluco-, and D-altropyranoside templates with an amphiphilic C-terminal hexadecapeptide aldehyde sequence. CD spectroscopy indicated that the choice of template has an effect on the overall structure of the carboprotein, as the altro-based carboprotein was found to be more alpha-helical than the corresponding galacto- and gluco-carboproteins. However, an influence on stability could not be detected in the present experiments, as the three carboproteins gave similar free energy of foldings (deltaG(F)H2O) and melting points in chemical and thermal denaturation experiments.     

Diphenyldichlorophosphonium trichloride-chlorine solvate 1:1, [PPh2Cl2]+Cl(3)(-).Cl2: an ionic form of diphenyltrichlorophosphorane. Crystal structures of [PPh2Cl2]+Cl(3)(-).Cl2 and [(PPh2Cl2)+]2[InCl5](2-)

An ionic form of diphenyltrichlorophosphorane, namely, diphenyldichlorophosphonium trichloride isolated as a dichlorine solvate (1), was obtained by treating PPh(2)Cl(3) with excess chlorine. The identity of this species was established by single-crystal X-ray analysis and (31)P, (1)H, and (35)Cl NMR and Raman spectra. Bis(diphenyldichlorophosphonium) pentachloroindate (2) was obtained by the reaction of diphenyltrichlorophosphorane with indium trichloride in dichloromethane for comparison purposes. Its identity was determined by (31)P NMR spectra and single-crystal X-ray analysis.     

Solvent-stabilized molecular capsules

Pyrrogallolarenes 2 were prepared by acid-catalyzed condensation of pyrrogallol with aldehydes. Compound 2a crystallizes from a methanol solution of quinuclidine hydrochloride to give a dimeric molecular capsule surrounding one disordered quinuclidinium cation. The molecules of 2a are connected by direct hydrogen bonds and by bridging methanol and water molecules. The chloride anion is positioned outside the capsule and is hydrogen bonded to the hydroxy groups of 2a. The shortest distance between the cation and anion was found to be 6.7 A. Crystallization of 2b from aqueous acetonitrile resulted in a dimeric capsule linked by a polar belt of 16 hydrogen bonding water molecules. Four acetonitrile molecules occupy the cavity of this dimeric capsule and assume two binding sites that differ in hydrogen bonding and electronic environment. Compounds 2 also form hydrogen-bonded dimeric molecular capsules in alcohols and aqueous acetonitrile solutions. These assemblies readily encapsulate tetramethylammonium, tetramethylphosphonium, quinuclidinium, and tropylium cations to give complexes stable on the NMR time scale at 233 K.     

A direct synthesis of 1-aryl- and 1-alkenylcyclopropylamines from aryl and alkenyl nitriles

The reaction of various aromatic nitriles with 1.1 equiv of Ti(Oi-Pr)(4) and 2.2 equiv of EtMgBr followed by addition of a Lewis acid gave 1-aryl cyclopropylamines in 43-76% yields. Under similar conditions, conjugated alkenenitriles afford 1-alkenylcyclopropylamines (42-65%).     

The biphasic virulence activities of gingipains: activation and inactivation of host proteins

Gingipains are trypsin-like cysteine proteinases produced by Porphyromonas gingivalis, a major causative bacterium of adult periodontitis. Rgps (HRgpA and RgpB) and Kgp are specific for -Arg-Xaa- and -Lys-Xaa- peptide bonds, respectively. HRgpA and Kgp are non-covalent complexes containing separate catalytic and adhesion/hemagglutinin domains, while RgpB has only a catalytic domain with a primary structure essentially identical to that of the cata-lytic subunit of HRgpA. The multiple virulence activities of gingipains are reviewed in view of the biphasic mechanisms: activation and inactivation of host proteins. Rgps enhanced vascular permeability through prekallikrein activation or direct bradykinin release in combination with Kgp. This Rgp action is potentially associated with gingival edema and crevicular fluid production. Rgps activate the blood coagulation system, leading to progression of inflammation and consequent alveolar bone loss in the periodontitis site. Rgps also activate protease-activated receptors and induce platelet aggregation, which, together with the coagulation-inducing activity, may explain an emerging link between periodontitis and cardiovascular disease. Kgp is the most potent fibrinogen/fibrin degrading enzyme of the three gingipains in human plasma, being involved in the bleeding tendency at the diseased gingiva. Gingipains stimulate expression of matrix metalloproteinases (MMPs) in fibroblasts and activate secreted latent MMPs that can destroy periodontal tissues. Gingipains degrade cytokines, components of the complement system and several receptors, including macrophage CD14, T cell CD4 and CD8, thus perturbing the host-defense systems and thereby facilitating sustained colonization of P. gingivalis. Gingipains are potent virulence factors of P. gingivalis, and in many regards their pathogenic activities constitute new mechanisms of bacterial virulence.     

What a difference ancillary thienyl makes: unexpected additional stabilization of the diruthenium(III,II) but not the diosmium(III,II) mixed-valent state in tetrazine ligand-bridged complexes

The Ru(2)(III,II) mixed-valent state is strongly stabilized in [(bpy)(2)Ru(mu-bttz)Ru(bpy)(2)](5+) (3(5+), bttz = 3,6-bis(2-thienyl)-1,2,4,5-tetrazine, as evident from lowered oxidation potentials and isolability, a strongly increased comproportionation constant K(c) = 10(16.6), and a high-energy intervalence charge transfer band at 10100 cm(-1). Curiously, no such effects were observed for the diosmium(III,II) analogue, whereas the related systems [(bpy)(2)M(mu-bmptz)M(bpy)(2)](5+), bmptz = 3,6-bis(4-methyl-2-pyridyl)-1,2,4,5-tetrazine, exhibit conventional behavior, i.e., a slightly higher K(c) value of the Os(2)(III,II) analogue. EPR signals were observed at 4 K for 3(5+) but not for the other mixed-valent species, and high-frequency (285 GHz) EPR was employed to study the diruthenium(II) radical complexes 2(3+) and 3(3+).     

High thermodynamic stability and extraordinary kinetic inertness of copper(II) complexes with 1,4,8,11-tetraazacyclotetradecane-1,8-bis(methylphosphonic acid): example of a rare isomerism between kinetically inert penta- and hexacoordinated copper(II) complexes

In an aqueous solution at room temperature, 1,4,8,11-tetraazacyclotetradecane-1,8-bis(methylphosphonic acid) (H(4)L(1)) and Cu(I) (I) form a pentacoordinated (pc) complex, pc-[Cu(L(1))](2-), exhibiting conformation I of the cyclam ring. At high temperature, the complex isomerises to a hexacoordinated isomer, trans-O,O-[Cu(L(1))](2-), with a trans-III conformation of the cyclam ring. In pc-[Cu(L(1))](2-), four ring nitrogen atoms and one phosphonate oxygen atom are arranged around Cu(I) (I) in a structure that is half-way between a trigonal bipyramid and a tetragonal pyramid, with one phosphonic acid group uncoordinated. In the trans-O,O-[Cu(L(1))](2-) isomer, the nitrogen atoms form a plane and the phosphonic acid groups are in a mutually trans configuration. A structurally very similar ligand, 4-methyl-1,4,8,11-tetraazacyclotetradecane-1,8-bis(methylphosphonic acid) (H(4)L(2)), forms an analogous pentacoordinated complex, pc-[Cu(L(2))](2-), at room temperature. However, the complex does not isomerise to the octahedral complex analogous to trans-O,O-[Cu(L(1))](2-). Because of the high thermodynamic stability of pc-[Cu(L(1))](2-), (logbeta=25.40(4), 25 degrees C, I=0.1 mol dm(-3) KNO(3)) and the formation of protonated species, Cu(I) (I) is fully complexed in acidic solution (-log [H(+)] approximately 3). Acid-assisted decomplexation of both of the isomers of [Cu(H(2)L(1))] takes place only after protonation of both uncoordinated oxygen atoms of each phosphonate moiety and at least one nitrogen atom of the cycle. The exceptional kinetic inertness of both isomers is illustrated by their half-lives tau(1/2)=19.7 min for pc-[Cu(H(2)L(1))] and tau(1/2) about seven months for trans-O,O-[Cu(H(2)L(1))] for decomplexation in 5 M HClO(4) at 25 degrees C. The mechanism of formation of pc-[Cu(L(1))](2-) is similar to those observed for other macrocyclic complexes.