Preferential Solvation of the Sodium Cation in Binary Mixtures of Tetrahydrofuran with Polyamines,in Relation with the Chelate Effect

The problem of determining the chelate effect brushes against methodological snags: the choice of concentration units, and of the appropriate standard states. We avoid these pitfalls by defining the chelate effect from measurements on bidentate ligands alone, without recourse to comparison with the corresponding unidentate ligands. Quantitation of the parameters extracted from the data is effected by three independent and mutually consistent procedures. Solvation of the Na⁺-cation by the polyamines follows the sequence: cadaverin < 1, 3-diaminopropane ? ethylene diamine ? diethylenetriamine. Entry of the first and of the second diamine molecule into the sodium coordination shell are independent and equiprobable steps: K₁ = K₃ and K₂ = K₄, within the accuracy of the measurements. For ethylene diamine, the values of K₁ and K₃ are in the range 1.0–1.5 and those for K₂ and K₄ are in the range 83–102: attachment of the second N-atom is considerably easier, by two orders of magnitude (chelate effect). The chelate effect is strongly reduced in cadaverin, with a longer hydrocarbon chain connecting the two amine functions.     

Noise-assisted traffic of spikes through neuronal junctions

The presence of noise, i.e., random fluctuations, in the nervous system raises at least two different questions. First, is there a constructive role noise can play for signal transmission in a neuron channel? Second, what is the advantage of the power spectra observed for the neuron activity to be shaped like 1/f(k)? To address these questions a simple stochastic model for a junction in neural spike traffic channels is presented. Side channel traffic enters main channel traffic depending on the spike rate of the latter one. The main channel traffic itself is triggered by various noise processes such as Poissonian noise or the zero crossings of Gaussian 1/f(k) noise whereas the variation of the exponent k gives rise to a maximum of the overall traffic efficiency. It is shown that the colored noise is superior to the Poissonian and, in certain cases, to deterministic, periodically ordered traffic. Further, if this periodicity itself is modulated by Gaussian noise with different spectral exponents k, then such modulation can lead to noise-assisted traffic as well. The model presented can also be used to consider car traffic at a junction between a main and a side road and to show how randomness can enhance the traffic efficiency in a network. (c) 2001 American Institute of Physics.     

Histochemical staining and quantification of dihydrolipoamide dehydrogenase diaphorase activity using blue native PAGE

Mammalian mitochondrial dihydrolipoamide dehydrogenase (DLDH, EC 1.8.1.4) catalyzes NAD(+)-dependent oxidation of dihydrolipoamide in vivo and can also act as a diaphorase catalyzing in vitro nicotinamide adenine dinucleotide (reduced form) (NADH)-dependent reduction of electron-accepting molecules such as ubiquinone and nitroblue tetrazolium (NBT). In this paper, we report a gel-based method for histochemical staining and quantification of DLDH diaphorase activity using blue native PAGE (BN-PAGE). Rat brain mitochondrial extracts, used as the source of DLDH, were resolved by nongradient BN-PAGE (9%), which was followed by diaphorase activity staining using NADH as the electron donor and NBT as the electron acceptor. It was shown that activity staining of DLDH diaphorase was both protein amount- and time-dependent. Moreover, this in-gel activity-staining method was demonstrated to be in good agreement with the conventional spectrophotometric method that measures DLDH dehydrogenase activity using dihydrolipoamide as the substrate. The method was applied to determine levels of DLDH diaphorase activity in several rat tissues other than the brain, and the results indicated a similar level of DLDH diaphorase activity for all the tissues examined. Finally, the effects of thiol-reactive reagents such as N-ethylmaleimide (NEM) and nitric oxide donors on DLDH diaphorase activity were evaluated, demonstrating that, with this method, DLDH diaphorase activity can be determined without having to remove these thiol-reactive reagents that may otherwise interfere with spectrophotometric measurement of DLDH dehydrogenase activity. The gel-based method can also be used as a means to isolate mitochondrial DLDH that is to be analyzed by mass spectral techniques in studying DLDH post-translational modifications.     

Fourier transform ion cyclotron resonance mass spectrometry of covalent adducts of proteins and 4-hydroxy-2-nonenal, a reactive end-product of lipid peroxidation

Covalent adduction of the model protein apomyoglobin by 4-hydroxy-2-nonenal, a reactive end-product of lipid peroxidation, was characterized by nanoelectrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (FTICR). The high mass resolving power and mass measurement accuracy of the instrument facilitated a detailed compositional analysis of the complex reaction product without the need for deconvolution and transformation to clearly show the pattern of adduction and component molecular weights. Our study has also demonstrated the value of electron capture dissociation over collision-induced dissociation for the tandem mass spectrometric determination of site modification for the 4-hydroxy-2-nonenal adduct of oxidized insulin B chain as an example. Figure FTICR allowed characterization of 4-hydroxy-2-nonenal (HNE)-modified apomyoglobin (an expanded spectrum of the +15 charge state is shown)