A highly sensitive method for quantification of myosin light chain phosphorylation by capillary isoelectric focusing with laser-induced fluorescence detection

Activation of myosin II by phosphorylation of the 20 kDa regulatory light chains (LC20) has been implicated in numerous contractile and motile events, e.g., smooth muscle contraction, cytokinesis, and cell migration. The ability to analyze LC20 phosphorylation in minute samples is critical to determine the importance of LC20 phosphorylation in diverse physiological processes. We have developed a method for the separation and quantification of unphosphorylated, monophosphorylated, and diphosphorylated LC20 with a detection limit of 1 pg (50 amol). LC20 is initially isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transblotted to a polyvinlyidene difluoride (PVDF) membrane. The region of the membrane containing the LC20 band (identified by electrophoresis of purified LC20 in a neighboring lane) is cut out and fluorescently labeled with Alexa Fluor 488 C5 maleimide. The labeled LC20 is eluted from the membrane with detergent and subjected to capillary isoelectric focusing (CIEF) to separate unphosphorylated, mono-, and diphosphorylated LC20, which are detected and quantified by laser-induced fluorescence (LIF). A linear relationship between log(peak area) and log(LC20 amount) is observed over the range of 50 amol-150 fmol. Quantification of LC20 phosphorylation by CIEF with LIF detection was compared with three commonly used methods with much lower levels of sensitivity: urea/glycerol-PAGE with Western blotting, phosphorylation by [gamma-32P]ATP with Cerenkov counting, and phosphorylation by [gamma-32P]ATP followed by SDS-PAGE, autoradiography, and scanning densitometry. All four methods gave very similar quantitative results, the major difference being that the new method exhibits 3000-fold enhanced sensitivity. This method is therefore applicable to quantitative analysis of phosphorylation of minute quantities of LC20.     

Zhang–Zhang Polynomials of Multiple Zigzag Chains Revisited: A Connection with the John–Sachs Theorem

Multiple zigzag chains Zm,n of length n and width m constitute an important class of regular graphene flakes of rectangular shape. The physical and chemical properties of these basic pericondensed benzenoids can be related to their various topological invariants, conveniently encoded as the coefficients of a combinatorial polynomial, usually referred to as the ZZ polynomial of multiple zigzag chains Zm,n. The current study reports a novel method for determination of these ZZ polynomials based on a hypothesized extension to John–Sachs theorem, used previously to enumerate Kekulé structures of various benzenoid hydrocarbons. We show that the ZZ polynomial of the Zm,n multiple zigzag chain can be conveniently expressed as a determinant of a Toeplitz (or almost Toeplitz) matrix of size m2×m2 consisting of simple hypergeometric polynomials. The presented analysis can be extended to generalized multiple zigzag chains Zkm,n, i.e., derivatives of Zm,n with a single attached polyacene chain of length k. All presented formulas are accompanied by formal proofs. The developed theoretical machinery is applied for predicting aromaticity distribution patterns in large and infinite multiple zigzag chains Zm,n and for computing the distribution of spin densities in biradical states of finite multiple zigzag chains Zm,n.     

有限型的首次返回例外集的测度和维数北大核心CSCD

设M=(m_(ij))是一个b×b阶矩阵且m_(ij)∈{0,1},∑_(M)是矩阵M=(m_(ij))诱导产生的有限型,σ是其上左推移算子.本文主要研究的是有限型动力系统(∑_(M),σ)上的首次返回速度问题.令τ_(k)(x)是点x∈∑_(M)首次返回到包含x的k阶柱集时间,且E_(α,β)={x∈∑_(M):lim inf_(k→∞)(logτ_(k)(x))/k=α,lim sup_(k→∞)(logτ_(k)(x))/k=β}.我们证明了:当M是不可约矩阵时,对任意0≤α≤β≤+∞,集合E_(α,β)的Markov测度要么等于0要么等于1并且具有满的Hausdorff维数.     

Set controllability of Markov jump switching Boolean control networks and its applications

In this article, a novel method is proposed for investigating the set controllability of Markov jump switching Boolean control networks (MJSBCNs). Specifically, the switching signal is described as a discrete-time homogeneous Markov chain. By resorting to the expectation and switching indicator function, an expectation system is constructed. Based on the expectation system, a novel verifiable condition is established for solving the set reachability of MJSBCNs. With the newly obtained results on set reachability, a necessary and sufficient condition is further derived for the set controllability of MJSBCNs. The obtained results are applied to Boolean control networks with Markov jump time delays. Examples are demonstrated to justify the theoretical results.     

The nearest neighbor random walk on subspaces of a vector space and rate of convergence

LetX be the collection ofk-dimensional subspaces of ann-dimensional vector spaceV _n overGF(q). A metric may be defined onX by letting

Optimality of a Threshold Policy in theM/M/ queue with repeated vacations

Consider anM/M/1 queueing system with server vacations where the server is turned off as soon as the queue gets empty. We assume that the vacation durations form a sequence of i.i.d. random variables with exponential distribution. At the end of a vacation period, the server may either be turned on if the queue is non empty or take another vacation. The following costs are incurred: a holding cost ofh per unit of time and per customer in the system and a fixed cost of each time the server is turned on. We show that there exists a threshold policy that minimizes the long-run average cost criterion. The approach we use was first proposed in Blanc et al. (1990) and enables us to determine explicitly the optimal threshold and the optimal long-run average cost in terms of the model parameters.     

On two--block--factor sequences and one--dependence

The distributions of two--block--factors arising from i.i.d. sequences are observed to coincide with the distributions of the superdiagonals of jointly exchangeable and dissociated arrays . An inequality for superdiagonal probabilities of the arrays is presented. It provides, together with the observation, a simple proof of the fact that a special one--dependent Markov sequence of Aaronson, Gilat and Keane (1992) is not a two--block factor.

     

The matched queueing network PH/M/c→oPH/PH/1

We consider a matched queueing network PH/M/coPH/PH/1, where the output of the first subsystem PH/M/c is regarded as one of the inputs of the second subsystem and matched with another input of PH-renewal process, and they are served by a single server with PH-service-distribution. The stationary state probabilities, the distribution of the virtual waiting times in the two subsystems, and the distribution of the total virtual waiting time are derived.The work was supported by the Max-Planck-Gesellschaft, Germany, and the Chinese Academy ofSciences.     

Random walk on distant mesh points Monte Carlo methods

A new technique for obtaining Monte Carlo algorithms based on the Markov chains with a finite number of states is suggested. Instead of the classical random walk on neighboring mesh points, a general way of constructing Monte Carlo algorithms that could be called random walk on distant mesh points is considered. It is applied to solve boundary value problems. The numerical examples indicate that the new methods are less laborious and therefore more efficient.In conclusion, we mention that all Monte Carlo algorithms are parallel and could be easily realized on parallel computers.     

Renormalization group for Markov chains and application to metastability

In this paper we introduce a new renormalization group method for the study of the long-time behavior of Markov chains with finite state space and with transition probabilities exponentially small in an external parameter. A general approach of metastability problems emerges from this analysis and is discussed in detail in the case of a two-dimensional Ising system at low temperature,