The Cytotoxic Effect of α-Synuclein Aggregates

Parkinson's disease is a neurodegenerative disorder involving a functional protein, α-synuclein, whose primary function is related to vesicle trafficking. However, α-synuclein is prone to form aggregates, and these inclusions, known as Lewy bodies, are the hallmark of Parkinson's disease. α-synuclein can alter its conformation and acquire aggregating capacity, forming aggregates containing β-sheets. This protein's pathogenic importance is based on its ability to form oligomers that impair synaptic transmission and neuronal function by increasing membrane permeability and altering homeostasis, generating a deleterious effect over cells. First, we establish that oligomers interfere with the mechanical properties of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) membrane, as demonstrated by nanoindentation curves. In contrast, nanoindentation revealed that the α-synuclein monomer's presence leads to a much more resistant lipid bilayer. Moreover, the oligomers’ interaction with cell membranes can promote lactate dehydrogenase (LDH) release, suggesting the activation of cytotoxic events.     

Initial-boundary value problems for incomplete singular perturbations of hyperbolic systems

The general problem studied has as a prototype the full non-linear Navier-Stokes equations for a slightly viscous compressible fluid including the heat transfer. The boundaries are of inflow-outflow type, i.e. non-characteristic, and the boundary conditions are the most general ones with any order of derivatives. It is assumed that the uniform Lopatinsky condition is satisfied. The goal is to prove uniform existence and boundedness of solution as the viscosity tends to zero and to justify the boundary layer asymptotics. The paper consists of two parts. In Part I the linear problem is studied. Here, uniform lower and higher order tangential estimates are derived and the existence of a solution is proved. The higher order estimates depend on the smoothness of coefficients; however this smoothness does not exceed the smoothness of the solution. In Part II the quasilinear problem is studied. It is assumed that for zero viscosity the overall initial-boundary value problem has a smooth solutionu ₀ in a time interval 0≦t≦T ₀. As a result the boundary laye, is weak and is uniformlyC ¹ bounded. This makes the linear theory applicable. an iteration scheme is set and proved to converge to the viscous solution. The convergence takes place for small viscosity and over the original time interval 0≦t≦T ₀.     

Disorder arising from the location of enantiomers in a twist-boat structure: 2,6-trans-2,4,6-tris(trichloromethyl)-1-oxa-3,5-dithian

Crystals of the title compound are orthorhombic, Pca2₁;a=27.586(4),b=10.509(3),c=11.080(2) Å,V=3212(1) ų,Z=8. The structure was solved by direct methods, from data collected at room temperature on an Enraf-Nonius CAD4 diffractometer, and refined by full-matrix least squares to a finalR value of 0.055 using 2043 reflections. One of the two independent molecules in the asymmetric unit is disordered. The disorder is manifested mainly between the oxygen atom and one sulfur atom and is caused by opposite enantiomers occupying corresponding positions in different unit cells. The endocyclic torsion angles in both independent molecules are very similar but the disposition of the heteroatoms on the twist-boat framework is different in the two molecules. Site occupancy of the two models on this framework in the disordered molecule is in the ratio 2179 with the smaller proportion being very similar to the ordered molecule and the remainder similar to that found in the normal molecule oftrans-4-chloro-2,4,6-tris(trichloromethyl)-1-oxa-3,5-dithian (Irving and Irving, 1988).     

High-throughput purification of combinatorial libraries I: a high-throughput purification system using an accelerated retention window approach

We have developed a high-throughput purification system to purify combinatorial libraries at a 50-100-mg scale with a throughput of 250 samples/instrument/day. We applied an accelerated retention window method to shorten the purification time and targeted one fraction per injection to simplify data tracking, lower QC workload, and simplify the postpurification processing. First, we determined the accurate retention time and peak height for all compounds using an eight-channel parallel LC/UV/MS system, and calculated the specific preparative HPLC conditions for individual compounds. The preparative HPLC conditions include the compound-specific gradient segment for individual compounds with a fixed gradient slope and the compound-specific UV or ELSD threshold for triggering a fraction collection device. A unique solvent composition or solvent strength was programmed for each compound in the preparative HPLC in order to elute all compounds at the same target time. Considering the possible deviation of the predicted retention time, a 1-min window around the target time was set to collect peaks above a threshold based on UV or ELSD detection. Dual column preparative instruments were used to maximize throughput. We have purified more than 500 000 druglike compounds using this system in the past 3 years. We report various components of this high-throughput purification system and some of our purification results.     

Studies with dithizone: Part XXI. The Relationship between pH12 and Molar Solubility

The effect of changes of ionic strength on the partition of an analytically important weak acid between a variety of aqueous and immiscible organic phases has been studied with respect to new measurements on dithizone (3-mercapto-1,5-diphenylformazan) and di(p-tolyl)thiocarbazone. A linear relationship has been deduced between the pH at which 50% extraction takes place and S_r,o (the solubility of the reactant in the organic phase), and confirmed by measurements of pH_{$\text{1}\text{2}$}, and S_r,o for a large number of organic solvents. The solubilities of the reagents in water, S_r, are also reported. The form of the relationship between pH and log D, and between pH_{$\text{1}\text{2}$} and log S_r,o is the same whether a solution of dithizone consists of a single species or a mixture of thiol and thione forms in tautomeric equilibrium.     

The determination of small amounts of long-chain tertiary alkylamines and quarternary ammonium salts

Bromocresol green has been investigated as an alternative to ammonium erdmannate for the absorptiometric determination of small amounts of long-chain alkylamines and quaternary ammonium salts in aqueous solution in a solvent extraction procedure. The molecular absorption coefficient of the reagent at 615 nm is about 3 times that of the erdmannate ion at 353 nm; visual methods of comparison are simplified by the blue colour of the bromocresol green anion The sensitivity of both the bromocresol green and the erdmannate method may be increased by stripping the coloured reagents into a small volume of aqueous solution. Both methods are free of interference from iron, but the present procedure has the disadvantage of a blank which depends both on the pH and on the concentration of excess reagent.     

Formation and structure of a novel enediyne-RNA base covalent adduct

The structure of an unusual covalent adduct formed by thiol-activated neocarzinostatin chromophore (NCS-chrom) and a RNA-DNA hybrid having an overhang of four unpaired residues at the 3'-end of the RNA strand has been elucidated by MS and NMR spectroscopic analyses. Unlike previously characterized adducts formed by NCS-chrom on the sugar residue of the DNA target, this adduct has been found to be on one of the uracil bases in the RNA overhang. Covalent linkage is between C-6 of the post-activated NCS-chrom and C-5 of the uracil. A novel mechanism involving adduction of the NCS-chrom C-6 radical, generated by 2-mercaptoethanol activation, to C-5 of the uracil at the U9 position of the RNA 11-mer, oxidation by dioxygen, reduction by the thiol, and subsequent dehydration is proposed for adduct formation.     

The organic chemistry of deoxynucleosides and deoxynucleotides

The organic chemistry of the naturally occurring deoxynucleosides and their phosphate esters is reviewed.