Compact disk (CD)-shaped device for single cell isolation and PCR of a specific gene in the isolated cell

For immediate discrimination among isolated cells we propose a novel device and technique for isolation of cells and sequential detection of specific gene(s) within them by polymerase chain reaction (PCR). In this study, we isolated Salmonella enterica cells and detected the Salmonella-specific invA gene from isolated cells by PCR on a compact disk (CD)-shaped device. This device enabled liquid flow by centrifugal force without a micro pump, and was fabricated from silicon wafer and glass to avoid evaporation of a small amount of reagent. One device has 24 microchannels, and 313 microchambers integrated on each microchannel. One microliter of PCR mixture containing cells was separated into microchambers on the device at 5000 rpm for 30 s. Each microchamber contained approximately 1.5 nL PCR mixture. A Poisson distribution of S. enterica cells was observed for different densities of cell suspension. At 200 cells μL^?1 of S. enterica or less, isolated single cells could be determined on the device by amplification of DNA of the invA gene; at 400 cells μL^?1, chambers containing no, one, two, or three cells could be determined on the device. Selective detection of S. enterica was achieved by PCR from a mixture of S. enterica and Esherichia coli on the CD-shaped device.     

Molecular simulation study with complex models of the carbohydrate binding module of Cel6A and the cellulose Iα crystal

A computer docking study was carried out on the (110) crystal surface of the cellulose Iα crystal model for the carbohydrate binding module (CBM) of cellobiohydrolase Cel6A, which is produced by the filamentous fungus Trichoderma reesei. Three-dimensional structures of the CBM were constructed by the homology modeling method using the Cel7A CBM, which is another cellobiohydrolase from T. reesei, as a template, and refined by molecular dynamics calculations in the solution state. Among the three models tested, those with three disulfide bonds were selected for a docking analysis. The binding free energy maps represented changes in non-covalent interactions and solvation free energies with respect to the CBM position. These indicated two minimum positions within the unit cell for both the parallel and antiparallel orientation modes of the CBM with respect to the cellulose fiber axis. Molecular dynamics calculations under an explicit solvent system were performed for the four complex models derived from the minimum positions of the binding free energy maps. The complex models with CBM in the parallel orientation had the lowest binding energies.     

Fundamental solution of a higher step Grushin type operator

We examine a class of Grushin type operators _Pk$P_k$ where k∈_N0$k\in {\mathbb{N}}_0$ defined in (1.1). The operators _Pk$P_k$ are non-elliptic and degenerate on a sub-manifold of R^N+?${\mathbb{R}}^{N+?}$. Geometrically they arise via a submersion from a sub-Laplace operator on a nilpotent Lie group of step k+1$k+1$. We explain the geometric framework and prove some analytic properties such as essential self-adjointness. The main purpose of the paper is to give an explicit expression of the fundamental solution of _Pk$P_k$. Our methods rely on an appropriate change of coordinates and involve the theory of Bessel and modified Bessel functions together with Weber's second exponential integral.     

Pressure‐sensitive adhesive utilizing molecular interactions between thymine and adenine

A simple pressure‐sensitive adhesion (PSA) system incorporating noncovalent interaction between thymine and adenine is presented. A copolymer having thymine moieties is combined with a low‐molecular‐weight bifunctional adenine cross‐linker. Molecular interactions caused by multiple hydrogen bonds between the thymine and adenine units are evaluated by FT‐IR spectral measurement. Mechanical properties of the PSA are examined by stress–strain curves and dynamic mechanical analysis. As the number of adenine cross‐linkers increases, Young's modulus increases from 0.24 to 3.0 MPa, and the glass transition temperature increases. Furthermore, it is found that the PSAs have adequate adhesive property from their shear strength test. Heat treatment at 80 °C is effective for reinforcement because of interchange of the hydrogen bonds. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2016 , 54, 1332‐1338     

Complete classification of pseudo <Emphasis Type="Italic">H</Emphasis>-type Lie algebras: I

Let \({\mathscr {N}}\) be a 2-step nilpotent Lie algebra endowed with a non-degenerate scalar product \(\langle .\,,.\rangle \), and let \({\mathscr {N}}=V\oplus _{\perp }Z\), where Z is the centre of the Lie algebra and V its orthogonal complement. We study classification of the Lie algebras for which the space V arises as a representation space of the Clifford algebra \({{\mathrm{{\mathrm{Cl}}}}}({\mathbb {R}}^{r,s})\), and the representation map \(J:{{\mathrm{{\mathrm{Cl}}}}}({\mathbb {R}}^{r,s})\rightarrow {{\mathrm{End}}}(V)\) is related to the Lie algebra structure by \(\langle J_zv,w\rangle =\langle z,[v,w]\rangle \) for all \(z\in {\mathbb {R}}^{r,s}\) and \(v,w\in V\). The classification depends on parameters r and s and is completed for the Clifford modules V having minimal possible dimension, that are not necessary irreducible. We find necessary conditions for the existence of a Lie algebra isomorphism according to the range of the integer parameters \(0\le r,s<\infty \). We present a constructive proof for the isomorphism maps for isomorphic Lie algebras and determine the class of non-isomorphic Lie algebras.     

Zeta regularized determinant of the Laplacian for classes of spherical space forms

We derive the spectral zeta function in terms of certain Dirichlet series for a variety of spherical space forms _MG$M_G$. Extending results in [C. Nash, D. O’Connor, Determinants of Laplacians on lens spaces, J. Math. Phys. 36 (3) (1995) 1462–1505] the zeta-regularized determinant of the Laplacian on _MG$M_G$ is obtained explicitly from these formulas. In particular, our method applies to manifolds of dimension higher than 3 and it includes the case where G$G$ arises from the dihedral group of order 2m$2m$. As a crucial ingredient in our analysis we determine the dimension of eigenspaces of the Laplacian in form of some combinatorial quantities for various infinite classes of manifolds from the explicit form of the generating function in [A. Ikeda, On the spectrum of a Riemannian manifold of positive constant curvature, Osaka J. Math. 17 (1980) 75–93].     

Synthesis of functional proteins by mixing peptide motifs

Here, we describe a synthetic approach for generating artificial proteins by the assemblage of naturally occurring peptide motifs. Two motifs respectively related to apoptosis induction and protein transduction were encrypted into different reading frames of an artificial gene (microgene), which was then polymerized; random frame shifts at the junctions between the microgene units yielded combinatorial polymers of three reading frames. Among the proteins created, #284 was found to penetrate through cell membranes and exert a strong apoptotic effect on several cancer cell lines. Because a simple linkage of these motifs was not sufficient to construct a bifunctional peptide, and the successful reconstitution was dependent on how they were joined together, the combinatorial strategy is important for reconstituting functions from mixtures of motifs. This microgene-based approach represents a novel system for creating proteins with desired functions.