Fast electrophoretic analysis of individual mitochondria using microchip capillary electrophoresis with laser induced fluorescence detection

The analysis of mitochondria by capillary electrophoresis usually takes longer than 20 min per replicate which may compromise the quality of the mitochondria due to degradation. In addition, low sample consumption may be beneficial in the analysis of rare or difficult samples. In this report, we demonstrate the ability to analyze individual mitochondrial events in picoliter-volume samples (approximately 80 pL) taken from a bovine liver preparation using microchip capillary electrophoresis with laser-induced fluorescence detection (micro-chip CE-LIF). Using a commercial "double-T" glass microchip, the sample was electrokinetically loaded in the "double-T" intersection and then subjected to electrophoretic separation along the main separation channel. In order to decrease interactions of mitochondria with channel walls during the analysis, poly(vinyl alcohol) was used as a dynamic coating. This procedure eliminates the need for complicated covalent surface modifications within the channels that were previously used in capillary electrophoresis methods. For analysis, mitochondria, isolated from bovine liver tissue, were selectively labelled using 10-nonyl acridine orange (NAO). The results consist of electropherograms where each mitochondrial event is a narrow spike (240 +/- 44 ms). While the spike intensity is representative of its NAO content, its migration time is used to calculate and describe its electrophoretic mobility, which is a property still largely unexplored for intracellular organelles. The five-fold decrease in separation time (4 min for microchip versus 20 min for capillary electrophoresis) makes microchip electrophoretic separations of organelles a faster, sensitive, low-sample volume alternative for the characterization of individual organelle properties and for investigations of subcellular heterogeneity.     

New methods for ligand-based virtual screening: use of data fusion and machine learning to enhance the effectiveness of similarity searching

Similarity searching using a single bioactive reference structure is a well-established technique for accessing chemical structure databases. This paper describes two extensions of the basic approach. First, we discuss the use of group fusion to combine the results of similarity searches when multiple reference structures are available. We demonstrate that this technique is notably more effective than conventional similarity searching in scaffold-hopping searches for structurally diverse sets of active molecules; conversely, the technique will do little to improve the search performance if the actives are structurally homogeneous. Second, we make the assumption that the nearest neighbors resulting from a similarity search, using a single bioactive reference structure, are also active and use this assumption to implement approximate forms of group fusion, substructural analysis, and binary kernel discrimination. This approach, called turbo similarity searching, is notably more effective than conventional similarity searching.     

On-column labeling for capillary electrophoretic analysis of individual mitochondria directly sampled from tissue cross sections

This technical note reports on a new procedure to on-column-label organelles sampled from a tissue cross section into a fused silica capillary. These organelles are then analyzed by capillary electrophoresis with postcolumn laser-induced fluorescence detection. In this procedure, the fluorescent label does not come in contact with the tissue, which facilitates visualization of the sampled tissue cross section. In addition, on-column labeling allows for better control of the reaction time and fluorescent label concentrations. As a proof-of-principle, we show results of mitochondria from rat gastrocnemius muscle cross sections that were on-column-labeled with 10-N-nonyl acridine orange (NAO), a mitochondrion-specific probe, and compare them with results for NAO in-tissue labeling of the same tissue. The new organelle labeling procedure reported here may easily be extended to the analysis of individual organelles in other biological samples and may become a valuable tool in studies investigating the role of mitochondria in muscle aging and exercise physiology.      

Mechanical properties of mixed conducting La0.5Sr0.5Fe1−x Co x O3−δ (0≤x≤1) materials

Young’s modulus, strain–stress behavior, fracture strength, and fracture toughness of (0≤×≤1) materials have been investigated in the temperature range 20–1,000°C. Young’s moduli of and , measured by resonant ultrasound spectroscopy, were 130±1 and 133±3 GPa, respectively. The nonlinear stress–strain relationship observed by four-point bending at room temperature was inferred as a signature of ferroelastic behavior of the materials. Above the ferroelastic to paraelastic transition temperature, the materials showed elastic behavior, but due to high-temperature creep, a nonelastic respond reappeared above ∼800°C. The room temperature fracture strength measured by four-point bending was in the range 107–128 MPa. The corresponding fracture toughness of , measured by single edge V-notch beam method, was 1.16±0.12 MPa·m^1/2. The measured fracture strength and fracture toughness were observed to increase with increasing temperature. The fracture mode changed from intragranular at low temperature to intergranular at high temperature. Tensile stress gradient at the surface of the materials caused by a frozen-in gradient in the oxygen content during cooling was proposed to explain the low ambient temperature fracture strength and toughness.     

Rugged modules: The opposite of flatness

Relative notions of flatness are introduced as a mean to gauge the extent of the flatness of any given module. Every module is thus endowed with a flatness domain and, for every ring, the collection of flatness domains of all of its modules is a lattice with respect to class inclusion. This lattice, the flatness profile of the ring, allows us, in particular, to focus on modules which have a smallest flatness domain (namely, one consisting of all regular modules.) We establish that such modules exist over arbitrary rings and we call them Rugged Modules. Rings all of whose (cyclic) modules are rugged are shown to be precisely the von Neumann regular rings. We consider rings without a flatness middle class (i.e., rings for which modules must be either flat or rugged.) We obtain that, over a right Noetherian ring every left module is rugged or flat if and only if every right module is poor or injective if and only if R = S×T, where S is semisimple Artinian and T is either Morita equivalent to a right PCI-domain, or T is right Artinian whose Jacobson radical properly contains no nonzero ideals. Character modules serve to bridge results about flatness and injectivity profiles; in particular, connections between rugged and poor modules are explored. If R is a ring whose regular left modules are semisimple, then a right module M is rugged if and only if its character left module M⁺ is poor. Rugged Abelian groups are fully characterized and shown to coincide precisely with injectively poor and projectively poor Abelian groups. Also, in order to get a feel for the class of rugged modules over an arbitrary ring, we consider the homological ubiquity of rugged modules in the category of all modules in terms of the feasibility of rugged precovers and covers for arbitrary modules.     

On the Weak Order Ideal Associated to Linear Codes

In this work we study a weak order ideal associated with the coset leaders of a non-binary linear code. This set allows the incrementally computation of the coset leaders and the definitions of the set of leader codewords. This set of codewords has some nice properties related to the monotonicity of the weight compatible order on the generalized support of a vector in \(\mathbb {F}_q^n\) which allows to describe a test set, a trial set and the set of zero neighbours of a linear code in terms of the leader codewords.     

Covering morphisms

Let (R, m) be a two-dimensional regular local ring and let A be a finitely generated torsion-free R-module. If A is a complete module, then Dan Katz and Vijay Kodiyalam show A satisfies five conditions. They ask whether these five conditions are equivalent without assuming A to be complete. In a previous paper we determined all implications among these five conditions with one exception. In this paper, with an additional hypothesis on the units of R, we resolve the remaining case.     

Alternative rings of small order

We construct all the alternative (but not associative) algebras of dimension at most 5 over a perfect field. For any prime p, we show that there are fifteen alternative rings of prime power order pⁿ ,n≤5, which are not associative. None of these rings is nil. Just one has a unity.