Regulation of lithospermic acid B and shikonin production in Lithospermum erythrorhizon cell suspension cultures

Cell suspension cultures of Lithospermum erythrorhizon produced a large amount of lithospermic acid B, a caffeic acid tetramer, as well as shikonin derivatives (each ca. 10% of dry wt.) when cultured in shikonin production medium M-9. Various culture factors for increasing the production of lithospermic acid B were investigated. Lithospermic acid B production was inhibited by 2, 4-D or NH4+, whereas it was stimulated by Cu2+. These regulatory patterns were similar to those for the production of shikonin derivatives in these cell cultures, suggestive of close relations and similar metabolic regulation between the production of these compounds. Cultivation under light illumination, however, showed that these metabolisms were independently regulated. In particular, blue light showed a stimulatory effect on lithospermic acid B production, while shikonin production was strongly inhibited, indicative of an effective condition for lithospermic acid B production.     

Preparation of Secondary Carbinamines via Alkylation of N-Boryl Imines Generated from Partial Reduction of Nitriles with NABH4-Carboxylic Acid

Secondary carbinamines have been prepared from alkylation of N-boryl imines which were generated in situ from partial reduction of nitriles with sodium brohydride modified by carboxylic acid.     

Numerical visualization of free surface oscillation predicted with arbitrary Lagrangian-Eulerian method

A numerical prediction method has been proposed to predict non-linear free surface oscillation in a three-dimensional container. The fluid motions are numerically predicted with Navier-Stokes equations discretized in a Lagrangian scheme with sufficient numerical accuracy. The profile of a free surface is precisely represented with three-dimensional body-fitted coordinates (BFC), which are regenerated in each computational step on the basis of the arbitrary Lagrangian-Eulerian (ALE) formulation. The computational method was applied to non-linear sloshings and transitions from sloshing to swirling motions. The predicted free surface motions were visualized as sequential image files and animations to understand their dynamic futures     

Weighted scattering matrices of regular coverings of graphs

We give a decomposition formula for the determinant det(I ? U(λ)) of the weighted bond scattering matrix U(λ) of a regular covering of G. Furthermore, we define an L-function of G, and give a determinant expression of it. As a corollary, we express some determinant of the weighted bond scattering matrix of a regular covering of G by means of its L-functions.     

Zeta Functions of Graph Coverings

We give a decomposition formula for the zeta function of a group covering of a graph.     

Pipette tip biosensors for bacterial double-stranded DNA using bioluminescence induced by zinc finger luciferase

The authors describe a pipette type of biosensor for detecting target genes and using a zinc finger protein fused to luciferase (ZF luciferase). The ZF protein binds to a specific DNA sequence, and the target double-stranded (ds) DNA can be detected by monitoring the enzymatic activity of ZF luciferase. A small avidin-immobilized reaction plate is placed on a plastic pipette tip (referred to as Biologi tip). The dsDNA detection procedures are carried out by using a programmable dispensing robot equipped with a photodetector. These procedures include (a) the aspiration of an analyte to capture the biotinylated target dsDNA (a product of a polymerase chain reaction) on the small reaction plate inside the pipette tip, (b) the introduction of ZF luciferase and luciferin into the pipette tip, and (c) migration of the pipette tip to the detection port to measure bioluminescence on the small reaction plate. The emission originating from luciferase activity is observed on the reaction plate containing immobilized biotin-tagged target dsDNA, whereas plates containing non-target or biotinylated single-stranded DNA only do not yield a signal. The intensity of emission increases proportionally to the concentration of dsDNA, and the detection limit of the target dsDNA is as low as 62 pM. An actual genomic DNA sample from Escherichia coli O157 was successfully detected by this automatic analyzer using the Biologi tip equipped with a reaction plate. This indicates that this system has a large potential for practical applications, including in particular point-of-care analyses in hygiene control, food safety testing, and clinical diagnosis.

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Graphical abstract A pipette-type biosensor was developed to detect target genes using a luciferase-fused zinc finger protein, where a small NeutrAvidin-immobilized reaction plate was placed on the tip, and the biotinylated target double-stranded DNA was detected by monitoring the bound luciferase activity.

     

Parallel computation and numerical visualization for non-uniform particles included in gas and liquid flows

A parallel computation method has been proposed for the mixing and segregation of granular mixture included in gas and liquid flows. In this method, a three-dimensional (3D) computational volume is decomposed into multiple sub-blocks and their geometries are represented by 3D body-fitted coordinates. The fluid-particle interactions are treated by two types of models: a two-way model for liquid-solid flows and a one-way model in case of gas-solid flows. The computations of the particle motions in the multiple sub-blocks are executed simultaneously on the basis of the distinct element method (DEM). Since a graphic process is also executed as one of the parallel jobs, the particle distributions can be visualized during the computations. The computational method was applied to the gas-solid flows consisting of different diameters and densities in the horizontal and inclined cylinders rotating around their axes. From the comparison with the experimental results, nearly uniform mixing and particle segregation are successfully predicted in the oscillating liquid flows. In addition, it has been indicated that the particle pathline is very effective to visualize and understand the flow patterns of the particles with different properties. The result of the computations for the liquid-solid flows demonstrated that the vertical segregation of the non-uniform particles is reasonably reproduced.     

Impairment in S-phase entry of splenocytes of Parp-1 knockout mice

One immediate cellular response to DNA damage is the polyADP-ribosylation reaction by poly(ADP-ribose) polymerase-1 (Parp-1). The importance of Parp-1 has been established in many cellular processes, such as the maintenance of genomic stability, DNA repair and cell-death induction. Here, we established Parp-1^−/− mice of C57BL/6J congenic strain and characterized the role of Parp-1 in cell-cycle progression. In this study, we also improved a method to observe G0/G1 to S-phase transition of splenocytes and bone marrow cells prepared from mice. The cells were cultured and stimulated with mitogens (50 μM ionomycin/1 μM phorbol 12, 13-dibutyrate). We found that addition of a commercially available growth supportive reagent, BM Condimed ^RH1, greatly enhanced the transition of G0/G1 to the S-phase, which was determined by bromodeoxyuridine (BrdU) incorporation to DNA. Using this method, G0/G1 to the S-phase entry was measured using splenocytes derived from Parp-1^−/−, Parp-1^+/− and wild-type (Parp-1^+/+) mice. DNA synthesis in Parp-1^+/+ and Parp-1^+/− splenocytes started from day 1 after addition of mitogens, whereas that in Parp-1^−/− cells started from day 2. The peak of the S-phase was at day 2 in all genotypes and notably DNA synthesis in Parp-1^−/− cells was approximately halved compared to Parp-1^+/+ cells on day 2, 3 and 4. These results suggested that Parp-1 is involved in positive regulation of S-phase entry in quiescent mouse splenocytes.     

A New Proof of Bartholdi's Theorem

We give a new proof of Bartholdi's theorem for the Bartholdi zeta function of a graph. Supported by Grant-in-Aid for Science Research (C)     

Mechahism of the base promoted fragmentatiom of chloroacetylhydrazide

Experiments are described supporting the positions that the visible transient intermediate occurring during the fragmentation of chloroacetylhydrazide is the anion of acetyldiazene, and that the concurrent reduction is not a simple recombination reaction.