Synthesis of Unsymmetrically Substituted Hexahydroimidazo [4,5-d] Imidazole-2,5-diones and Elucidation of Reaction Pathways

We have reported the synthesis and structural elucidation of a series of hexahydroimidazo [4,5-d] imidazole-2,5-diones and hydantoins from glyoxal and substituted ureas.^1,2 The general equation of this condensation reaction and a few representative compounds: those reported, current and hypothetical, are depicted in Scheme 1.     

Neuronal activation by GPI-linked neuroligin-1 displayed in synthetic lipid bilayer membranes

We have characterized, in vitro, interactions between hippocampal neuronal cells and silica microbeads coated with synthetic, fluid, lipid bilayer membranes containing the glycosylphosphatidyl inositol (GPI)-linked extracellular domain of the postsynaptic membrane protein neuroligin-1. These bilayer-neuroligin-1 beads activated neuronal cells to form presynaptic nerve terminals at the point of contact in a manner similar to that observed for live PC12 cells, ectopically expressing the full length neuroligin-1. The synthetic membranes exhibited biological activity at neuroligin-1 densities of approximately 1 to 6 proteins/microm(2). Polyolycarbonate beads with neuroligin-1 covalently attached to the surface failed to activate neurons despite the fact that neuroligin-1 binding activity is preserved. This implies that a lipid membrane environment is likely to be essential for neuroligin-1 activity. This technique allows the study of isolated proteins in an environment that has physical properties resembling those of a cell surface; proteins can diffuse freely within the membrane, retain their in vivo orientations, and are in a nondenatured state. In addition, the synthetic membrane environment affords control over both lipid and protein composition. This technology is easily implemented and can be applied to a wide variety of cellular studies.     

Superoxide radical anion adduct of 5,5-dimethyl-1-pyrroline N-oxide (DMPO). 2. The thermodynamics of decay and EPR spectral properties

The formation of the superoxide radical anion (O2*-) adduct of the nitrone 5,5-dimethyl-1-pyrroline N-oxide (DMPO) as detected by electron paramagnetic resonance (EPR) spectroscopy is one of the most common techniques for O2*- detection in chemical and biological systems. However, the nature of DMPO-O2H has confounded spin-trapping investigators over the years, since there has been no independently synthesized DMPO-O2H to date. A density functional theory (DFT) approach was used to predict the isotropic hyperfine coupling constants arising from the N, beta-H, and gamma-H nuclei of DMPO-O2H using explicit interactions with water molecules as well as via a bulk dielectric effect employing the polarizable continuum model (PCM). Theoretical calculation on the thermodynamics of DMPO-O2H decay shows favorable intramolecular rearrangement to form a nitrosoaldehyde and a hydroxyl radical as products, consistent with experimental observations. Some pathways for the bimolecular decomposition of DMPO-O2H and DMPO-OH have also been computed.     

Structure of mercaptobiphenyl monolayers on mercury

The molecular-scale structure and phase behavior of single-component Langmuir films of 4'-methyl-4-mercaptobiphenyl (MMB) and 4'-perfluoromethyl-4-mercaptobiphenyl (FMMB) on mercury were studied using surface tensiometry, grazing incidence X-ray diffraction, and X-ray reflectivity. At low coverages, a condensed but in-plane disordered single layer of surface-parallel molecules is found for both compounds. At high coverages, both compounds exhibit in-plane-ordered phases of standing-up molecules. For MMB, the biphenyl core dominates the structure, yielding a centered-rectangular unit cell with an area A(x) of 21.8 A(2)/molecule, with molecules tilted by approximately 14 degrees from the surface normal in the nearest-neighbor direction, and a coherence length xi of >1000 A for the crystalline domains. For FMMB, the perfluoromethyl group dominates the structure, yielding a hexagonal unit cell with untilted molecules, an area A(x) of 24.2 A(2)/molecule, and a much smaller xi of approximately 110 A. The structure is discussed in comparison with self-assembled monolayers of MMB on crystalline Au(111) and similar-length alkanethiolate SAMs on Au(111) and on mercury. The differences in the structure are discussed and traced to the differences in the substrate's surface structure, and in the molecular cross section and rigidity.     

Characterization of amyloidogenic immunoglobulin light chains directly from serum by on-line immunoaffinity isolation

Primary systemic amyloidosis (AL) is characterized by the overproduction of immunoglobulin light chain proteins by a monoclonal, terminally differentiated B-lymphocyte or plasma cell clone. The free immunoglobulin light chains are deposited in an abnormal conformation as amyloid in a variety of organs in the body. The mechanism of amyloid formation is not well understood, but appears to be associated with some form of cleavage of the immunoglobulin light chain with subsequent aggregate formation. In an attempt to characterize the structure of amyloid-forming light chain proteins we developed an on-line immunoaffinity purification and subsequent characterization of free kappa and free lambda immunoglobulin light chains by electrospray ionization mass spectrometry. The methodology is totally automated and requires 20 micro L of serum. Mass spectral analysis of Bence Jones proteins under non-denaturing conditions was also utilized to examine the tertiary and quaternary structure of light chain proteins and clearly shows covalent dimer formation of lambda type light chain. This type of on-line assay may prove helpful in elucidating distinguishing features capable of discriminating AL from benign monoclonal gammopathies of undetermined significance as well as diagnosing AL.     

Theoretical study of the spin trapping of hydroxyl radical by cyclic nitrones: a density functional theory approach

The hydroxyl radical (*OH) is an important mediator of biological oxidative stress, and this has stimulated interest in its detection. 5,5-Dimethyl-1-pyrroline N-oxide (DMPO) and its alkoxycarbonyl and alkoxyphosphoryl analogues have been employed as spin traps for electron paramagnetic resonance (EPR) spectroscopic radical detection. Energies of optimized geometries of nitrones and their corresponding *OH adducts were calculated using density functional theory (DFT) at the B3LYP/6-31+G//B3LYP/6-31G level. Calculations predict that the trans adduct formation is favored in alkoxycarbonyl nitrones, while cis adducts with intramolecular H-bonding is favored for alkoxyphosphoryl nitrones. Addition of *OH to a phosphoryl-substituted nitrone is more exoergic than the carbonylated nitrones. Charge and spin densities on the nitrone spin traps were correlated with their rates of addition with *OH, and results show that the charge density on the nitronyl C, the site of *OH addition, is more positive in phosphorylated nitrones compared to DMPO and the alkoxycarbonyl nitrones. The dihedral angle between the beta-H and nitroxyl O bonds is smaller in phosphorylated nitrones, and that aspect appears to account for the longer half-lives of the spin adducts compared to those in DMPO and alkoxycarbonyl nitrones. Structures of nitrones with trifluoromethyl-, trifluoromethylcarbonyl-, methylsulfonyl-, trifluoromethylsulfonyl-, amido-, spiropentyl-, and spiroester substituents were optimized and their energies compared. Amido and spiroester nitrones were predicted to be the most suitable nitrones for spin trapping of *OH due to the similarity of their thermodynamic and electronic properties to those of alkoxyphosphoryl nitrones. Moreover, dimethoxyphosphoryl substitution at C-5 was found to be the most efficient substitution site for spin trapping of *OH, and their spin adducts are predicted to be the most stable of all of the isomeric forms.     

Absence of charge inversion on rodlike polyelectrolytes with excess divalent counterions

Filamentous viruses such as fd and M13 are highly charged rodlike polyelectrolytes. In this study, we employ fd virus to test the recent prediction of charge inversion [Nguyen, Rouzina, and Shklovskii, J. Chem. Phys. 112, 2562 (2000)]. Light scattering measurements show bundle formation and resolubilization of fd viruses when MgCl(2) was added from 0 to 600 mM. The effective charge of fd was studied by measuring their electrophoretic mobility using a filament tracking method uniquely suited for the system. Monte Carlo simulations were performed under canonical ensemble to predict the charge distribution around the rodlike virus. Charge inversion, which has been suggested theoretically to accompany with bundle resolubilization, was not observed in either experiments or simulations. A modified analysis of force balance is called upon to account for these new findings.     

Physicomechanical properties of nanocomposites based on cellulose nanofibre and natural rubber latex

Cellulose nanofibres (CNF) with diameter 10–60 nm were isolated from raw banana fibres by steam explosion process. These CNF were used as reinforcing elements in natural rubber (NR) latex along with cross linking agents to prepare nanocomposite films. The effect of CNF loading on the mechanical and dynamic mechanical (DMA) properties of NR/CNF nanocomposite was studied. The morphological, crystallographic and spectroscopic changes were also analyzed. Significant improvement of Young’s modulus and tensile strength was observed as a result of addition of CNF to the rubber matrix especially at higher CNF loading. DMA showed a change in the storage modulus of the rubber matrix upon addition of CNF which proves the reinforcing effect of CNF in the NR latex. A mechanism is suggested for the introduction of the Zn–cellulose complex and its three dimensional network as a result of the reaction between the cellulose and the Zinc metal which is originated during the composite formation.     

Application of PM-IRRAS to study thin films on industrial and environmental samples

Polarization modulation–infrared reflection absorption spectroscopy (PM-IRRAS) was employed to analyze two unique samples: (1) an industrially prepared alkoxysilane-pretreated aluminum alloy (AA6111) in the absence and presence of a ～600-nm-thick lubricant coating and (2) a chemical warfare agent simulant, triethyl phosphate (TEP), on glass. For the pretreated aluminum samples, PM-IRRAS spectra were analyzed for three distinct regions; the SiO stretching vibration around 1120 cm^?1, the NH₂ bending mode at ～1600 cm^?1 and the CH stretching region around 2900 cm^?1. Our results showed that increasing the curing temperature (from 55 to 100 °C) improved the overall extent of cross-linking within the siloxane network. In addition, the spectra of lubricant (top coating) and the underlying siloxane layer for the aluminum samples with lubricant were collected for the same sample. Our results show that the nature of the siloxane film remains intact and unaltered after deposition of the lubricant top-coat. For detection of TEP on glass, the band at 1268 cm^?1, corresponding to the P═O vibration, was monitored. A droplet of TEP solution in dichloromethane was deposited on glass. After solvent evaporation had occurred, the intensity of the P═O vibration band was used to construct calibration curves to determine the experimental limit of detection, which was found to be ～200 μg for TEP on glass.

Electrochemical studies on the interaction of l-cysteine with metallic copper in sulfuric acid

Amino acids have been widely used as green corrosion inhibitors for an array of metals. Considering its importance in corrosion chemistry, studies were undertaken with the objective to discovering the inhibitory effect of a sulfur-containing amino acid, l-cysteine, on copper in different concentrations of sulfuric acid (0.5, 1.0, and 1.5 M) at different temperatures. Techniques like the weight loss method, electrochemical impedance spectroscopy, potentiodynamic polarization (Tafel), and adsorption studies were employed. Results revealed that l-cysteine do offer an attractive inhibition efficiency. However, with an increase in the concentration of the inhibitor, corrosion rates decreased irrespective of the temperature gradients. This is due to surface adsorption of the inhibitor molecules on the metal which has contributed to a decreased double-layer capacitance and increased polarization resistance. With the increase in the concentration of the medium, the corrosion rate was also enhanced and this is due the liberation of a high quantum of H⁺ ions. Based on the results of Tafel polarization studies, it is evident that the amino acid, l-cysteine, could act as a mixed type inhibitor. The importance of l-cysteine in the corrosion of copper metal has been highlighted in this paper.