Immunogenicity of Foot-and-Mouth Disease Virus Dendrimer Peptides: Need for a T-Cell Epitope and Ability to Elicit Heterotypic Responses

An approach based on a dendrimer display of B- and T-cell epitopes relevant for antibody induction has been shown to be effective as a foot-and-mouth disease (FMD) vaccine. B₂T dendrimers combining two copies of the major FMD virus (FMDV) type O B-cell epitope (capsid proteinVP1 (140–158)) covalently linked to a heterotypic T-cell epitope from non-structural protein 3A (21–35), henceforth B₂T-3A, has previously been shown to elicit high neutralizing antibody (nAb) titers and IFN-γ-producing cells in both mice and pigs. Here, we provide evidence that the B- and T-cell epitopes need to be tethered to a single molecular platform for successful T-cell help, leading to efficient nAb induction in mice. In addition, mice immunized with a non-covalent mixture of B₂T-3A dendrimers containing the B-cell epitopes of FMDV types O and C induced similarly high nAb levels against both serotypes, opening the way for a multivalent vaccine platform against a variety of serologically different FMDVs. These findings are relevant for the design of vaccine strategies based on B- and T-cell epitope combinations.     

Chirality and Spectroscopic Changes Induced by the Recognition of Ethyl 5-Amino-2-methyl-1,2-dihydro-3-phenylpyrido[3,4-b]pyrazin-7-yl Carbamate Analogs by Tubulin

The two chiral isomers of ethyl 5-amino-2-methyl-l,2-di-hydro-3-phenylpyrido[3,4-b]pyrazin-7-yl carbamate, NSC 613863 (R-isomer)-(+) and NSC 613862 (S-isomer)-(-) (CI980) and the three achiral analogs NSC 330770 (2-de-methylated analog A), NSC 337238 and C179 are potent microtubule inhibitors. These ligands interact with tubulin overlapping the colchicine binding site. This study addresses the effects of recognition by tubulin on the conformational properties of the ligands. The near-UV CD (circular dichroism) band of the R-isomer was suppressed, while that of the S-isomer displayed a more intense negative band when these compounds were bound to tubulin. Interestingly, the three other initially achiral compounds became optically active upon binding to tubulin; particularly, analog A exhibited a negative CD band on the order of magnitude of chiral compounds. The CD changes are reversible, highly specific and actually permit measuring the binding of the ligands by tubulin. These CD changes are compatible with the deformation of the bound ligands. Fluorescence emission is strongly enhanced and blue shifted upon binding to tubulin. Water among a solvent series had a specific solvent effect, except on the 1,2-dehydro analogs NSC 337238 and C179, suggesting hydrogen bonding to Nl. The emission of tubulin-bound R-isomer, S-isomer and analog A could be mimicked by solvent viscosity, supporting the notion that the intramolecular rotation between the pyridopyrazine and phenyl rings is frozen upon binding.     

A spectral mapping theorem for perturbed strongly continuous semigroups

The second author was supported by a F.P.I. grant from Spanish Ministerio de Educación y Ciencia     

A note on a theorem of F. Browder

A particular case of a mathematical theorem of F. Browder on the behavior of the fixed point set of a mapping under variations of a parameter has recently found applications in programming theory in connection with the abstract (non-linear) complementarity problem (see Eaves, [2, 3]). Two relevant extensions of Browder's result are provided: The first asserts that, under smoothness assumptions, the connected set of fixed points one gets from Browder's theorem is “generically” an arc; the second gives a generalization to the case where the mapping is an upper hemicontinuous contractible valued correspondence.     

Polycrystalline silicon films obtained by hot-wire chemical vapour deposition

Silicon films were deposited at moderate substrate temperatures (280–500° C) from pure silane and a silane-hydrogen mixture (10% SiH₄, 90% H₂) in a hotwire CVD reactor. The morphology, structure and composition of the samples were studied with scanning electron microscopy, transmission electron microscopy, transmission electron diffraction, X-ray diffraction, Raman spectroscopy and secondary ion mass spectrometry. The sample deposited at 500° C with pure silane has an amorphous structure, whereas the samples obtained from silane diluted in hydrogen have a polycrystalline structure, even that grown at the lowest temperature (280° C). Polycrystalline samples have a columnar structure with 0.3–1 m crystallite sizes with preferential orientation in [220] direction. Deposition rates depend on the filament-substrate distance and range from 9.5 to 37 Å/s for the polycrystalline samples. The high quality of the polycrystalline samples obtained makes the hot-wire technique very promising. Moreover, it is expected to be easily scaled up for applications to large-area optoelectronic devices and to photovoltaic solar cells.     

Stabilization of solutions of the filtration equation with absorption and non-linear flux

This paper is primarily concerned with the large time behaviour of solutions of the initial boundary value problem $$\begin{gathered} u_t = \Delta \phi (u) - \varphi (x,u)in\Omega \times (0,\infty ) \hfill \\ - \frac{{\partial \phi (u)}}{{\partial \eta }} \in \beta (u)on\partial \Omega \times (0,\infty ) \hfill \\ u(x,0) = u_0 (x)in\Omega . \hfill \\ \end{gathered} $$ Problems of this sort arise in a number of areas of science; for instance, in models for gas or fluid flows in porous media and for the spread of certain biological populations.     

Liquid Chromatography-Mass Spectrometry Hyphenation for Exhaustive and Unambiguous Characterization of Polyoxyethylene Surfactants

A rapid, specific and reliable high performance liquid chromatographic (HPLC) assay of sildenafil in pharmaceutical dosage forms has been developed and evaluated. Reversed phase chromatography was conducted using a mobile phase of methanol: water: acetonitrile (60:20:20) v/v/v, pH 6.1, 0.1% glacial acetic acid, and detection at λ 290 nm. The recovery and coefficient of variation from six tablets containing 50 mg of sildenafil were 100.90% and 0.45% respectively. Replicate regression analysis of three standard plots in the concentration range (0.01–0.2) μg mL^?1 obtained on three different days gave a correlation coefficient >0.999 and the coefficient of variation of the slopes <1.5%. The assay was precise within day and between days as indicated by ANOVA test. It is suggested that the proposed HPLC method should be used for routine quality control and dosage form assay of sildenafil citrate. The proposed method was also used to study the stability of sildenafil citrate in different dosage forms of the drug.     

Synthesis of new ether glycerophospholipids structurally related to modulator

A series of new glycerophospholipids, bearing a short-chain carboxylic acid in position sn-1 and phosphocholine or phosphoserine in postion sn-3 of glycerol, have been prepared in good overall yields. Compound 11, 1-0-(6-carboxyhexyl)-sn-glycero-3-phosphoserine, a strict analog of the structure proposed for modulator, has been synthesized in a stereoselective way from (R)-1,2-isopropylideneglycerol 1.     

Enhancement of the fluorescence of the zinc-morin complex by a non-ionic surfactant

A method is described for the fluorimetric determination of zinc, based on formation of a zinc-morin complex in the presence of a non-ionic surfactant. The complex has practically no fluorescence in the absence of surfactant, but the addition of Genapol PF-20 (non-ionic surfactant, ethylene oxide-propylene oxide condensate) makes possible the fluorimetric determination of low concentrations of zinc as it enhances the fluorescence of the complex about 75-fold. Maximum fluorescence is produced at pH 4.7 +/- 0.2 (acetic acid-acetate buffer), with 1.5% surfactant and 0.009% morin. The fluorescence is excited at 433 nm and measured at 503 nm. The calibration graph is linear up to 150 ng/ml zinc concentration and the detection limit is 3 ng/ml. The relative standard deviation (11 replicates) is 2.4% for zinc at 20 ng/ml concentration and 1.7% for 100 ng/ml. Of 29 ions studied, Al(3+), Be(2+), Zr(4+) and Cd(2+) strongly increase the fluorescence of the system, and Fe(3+), Ni(2+), Cu(2+), Ti(IV) and Co(2+) decrease the fluorescence signal.