Red blood cells do not attenuate the SPCE fluorescence in surface assays

We describe the positive effect of surface plasmon-coupled fluorescence emission (SPCE) on the detection of a signal from a surface immunoassay in highly absorbing or/and scattering samples. A model immunoassay using fluorescently labeled anti-rabbit antibodies that bind to rabbit immunoglobulin on a silver surface was performed, and the signal was detected in the presence of various highly absorbing and/or scattering solutions or suspensions, such as hemoglobin solution, plastic beads, and red blood cells. The results showed that a highly absorbing solution consisting of small molecules (dye, hemoglobin) attenuates the SPCE signal approximately 2-3-fold. In contrast, suspensions with the same absorption containing large particles (large beads, red blood cell suspension) attenuate the SPCE signal only slightly, approximately 5-10%. Also, a suspension of large undyed, highly scattering beads does not reduce the SPCE signal. The effects on the immunoassay signal of the sample background absorption and scattering, the size of the background particles, and the geometry of the experimental set-up are discussed. We believe that SPCE is a promising technique in the development of biosensors utilized for surface-based assays, as well as any assays performed directly in highly absorbing and/or scattering solutions without washing or separation procedures. Figure Red blood cells (unlike hemoglobin) do not attenuate the SPCE fluorescence in surface assays.     

A multiplicity theorem for problems with the p-Laplacian

We consider a nonlinear elliptic problem driven by the p-Laplacian, with a parameter λ∈R and a nonlinearity exhibiting a superlinear behavior both at zero and at infinity. We show that if the parameter λ is bigger than λ₂=the second eigenvalue of , then the problem has at least three nontrivial solutions. Our approach combines the method of upper-lower solutions with variational techniques involving the Second Deformation Theorem. The multiplicity result that we prove extends an earlier semilinear (i.e. p=2) result due to Struwe [M. Struwe, Variational Methods, Springer-Verlag, Berlin, 1990].     

Nonlinear surface waves in soft, weakly compressible elastic media

Nonlinear surface waves in soft, weakly compressible elastic media are investigated theoretically, with a focus on propagation in tissue-like media. The model is obtained as a limiting case of the theory developed by Zabolotskaya [J. Acoust. Soc. Am. 91, 2569-2575 (1992)] for nonlinear surface waves in arbitrary isotropic elastic media, and it is consistent with the results obtained by Fu and Devenish [Q. J. Mech. Appl. Math. 49, 65-80 (1996)] for incompressible isotropic elastic media. In particular, the quadratic nonlinearity is found to be independent of the third-order elastic constants of the medium, and it is inversely proportional to the shear modulus. The Gol'dberg number characterizing the degree of waveform distortion due to quadratic nonlinearity is proportional to the square root of the shear modulus and inversely proportional to the shear viscosity. Simulations are presented for propagation in tissue-like media.     

Probing transient Hoogsteen hydrogen bonds in canonical duplex DNA using NMR relaxation dispersion and single-atom substitution

Nucleic acids transiently morph into alternative conformations that can be difficult to characterize at the atomic level by conventional methods because they exist for too little time and in too little abundance. We recently reported evidence for transient Hoogsteen (HG) base pairs in canonical B-DNA based on NMR carbon relaxation dispersion. While the carbon chemical shifts measured for the transient state were consistent with a syn orientation for the purine base, as expected for A(syn)?T(anti) and G(syn)?C(+)(anti) HG base pairing, HG type hydrogen bonding could only be inferred indirectly. Here, we develop two independent approaches for directly probing transient changes in N-H···N hydrogen bonds and apply them to the characterization of transient Hoogsteen type hydrogen bonds in canonical duplex DNA. The first approach takes advantage of the strong dependence of the imino nitrogen chemical shift on hydrogen bonding and involves measurement of R(1ρ) relaxation dispersion for the hydrogen-bond donor imino nitrogens in G and T residues. In the second approach, we assess the consequence of substituting the hydrogen-bond acceptor nitrogen (N7) with a carbon (C7H7) on both carbon and nitrogen relaxation dispersion data. Together, these data allow us to obtain direct evidence for transient Hoogsteen base pairs that are stabilized by N-H···N type hydrogen bonds in canonical duplex DNA. The methods introduced here greatly expand the utility of NMR in the structural characterization of transient states in nucleic acids.     

Changes in intracellular potassium and sodium content of 2-cell mouse embryos induced by exposition to vitrification concentrations of ethylene glycol

Intracellular concentration of potassium and sodium in two-cell mouse embryos in G1/S phase after exposition to vitrification solutions containing ethylene glycol (EG) and sucrose or after incubation in Dulbecco solution were measured by electron probe microanalysis (EPMA). The embryos at room temperature were treated in 10 percent EG for 10 min, transferred into mixture of EG and 1.0 M sucrose in ratio of 3:7 (v/v) for 3 min, then to 0.5 M sucrose for 10 min followed by washing the cells with Dulbecco;s solution for 10 min prior to analysis. The cytoplasmic concentration of potassium and sodium in controlled untreated with EG embryos were in a range of 116-130 mM of potassium and 120 mM of sodium, with good concordance in two identical experiments. After exposition that mimicked vitrification protocols, the intracellular potassium dropped almost two-three-fold (47 + 3 mM in one experiment and to 70 mM in the second experiment. The intracellular sodium concentration also decreased two-fold in range 60-70 mM after treatment with EG. Possible mechanisms of changes in the intracellular elemental concentrations including the high intracellular sodium observed in intact embryos are discussed.     

Stereoselective synthesis of enantiopure cyclic α‐aminophosphonic acids: Direct observation of inversion at phosphorus in phosphonate ester silyldealkylation by bromotrimethylsilane

The paper describes a simple, direct synthesis of enantiopure cyclic α‐aminophosphonic acids on the basis of silyldealkylation and followed by hydrolysis of the parent diastereoisomeric cyclic 1,4,2‐oxazaphosphorines, which have been obtained by intramolecular stereospecific nucleophilic addition of phosphites to imines. In this approach, the high diastereoselectivity of the stereocontrolling penultimate step is preserved by conversion of the intermediate ester to the final phosphonic acid under very mild, nonracemizing conditions. © 2008 Wiley Periodicals, Inc. Heteroatom Chem 19:575–582, 2008; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/hc.20480     

Polyacrylamide gel electrophoresis of fluorophore-labeled hyaluronan and chondroitin sulfate disaccharides: application to the analysis in cells and tissues

This report describes a new formulation of polyacrylamide gel electrophoresis of fluorophore-labeled saccharides (PAGEFS) for the analysis of hyaluronan (HA) and chondroitin sulfate (CS) Delta-disaccharides. PAGEFS relies on derivatization of reducing ends of HA- and the variously sulfated CS-derived Delta-disaccharides with 2-aminoacridone (AMAC), followed by electrophoresis under optimized buffer conditions (Tris-borate and Tris-HCl) and on polyacrylamide gels (25% T/3.75% C). The method was applied to the analysis of glycosaminoglycans (GAGs) from the human umbilical cord tissue and GAGs isolated from human aortic smooth muscle cell cultures. The obtained results were in agreement with those obtained after an analysis with high-performance liquid chromatography (HPLC). On the basis of these results, PAGEFS is a rapid and sensitive method for the analysis of the total amount of HA- and CS-derived disaccharides, as it allows analyzing 20 samples in minigels in one run and provides quantitation with relatively high sensitivity (less than 25 pmol per disaccharide). In addition, PAGEFS overcomes the lack of commercial gels described previously for the separation of AMAC-labeled disaccharides. Therefore, the method proposed here is an economic and useful tool for a fast screening of GAGs in biological samples, particularly when a high number of samples should be analyzed.     

Modifications of the equation for gas bubble dynamics in a soft elastic medium

A model equation for the oscillation of a pressurized gas bubble in a nonlinear incompressible elastic medium [Emelianov et al., J. Acoust. Soc. Am. 115, 581 (2004)] is extended to include effects of surface tension, viscosity, weak compressibility, and confinement by an elastic shell. The significance of this work is that starting from first principles, the full nonlinearity of the incompressible elastic medium surrounding the bubble and forming its shell is taken into account. Measurements of equilibrium radius as a function of external pressure for a gas bubble in a tissue-like gel are also presented. A general approach to including hysteresis is also discussed.