Two-dimensional network formation of cardiac myocytes in agar microculture chip with 1480 nm infrared laser photo-thermal etching

We have developed a new method that enables agar microstructures to be used to cultivate cells and that allows cell network patterns to be controlled. The method makes use of non-contact three-dimensional photo-thermal etching with a 1480 nm infrared focused laser beam, which is strongly absorbed by water and agar gel, to form the shapes of agar microstructures. It allows microstructures to be easily formed in an agar layer within a few minutes, with cell-culture holes formed by the spot heating of a 100 mW laser and tunnels by the tracing of a 100 microm s(-1), 40 mW laser. We cultivated rat cardiac myocytes in adjacent microstructures and observed synchronized beating in them 90 min after they had made physical contact. Our results indicate that the system can make and use microstructures for cell-network cultivation in a minimal amount of time without any expensive microfabrication facilities or complicated procedures.     

Structure-specificity relationship of cardiac glycosides as a substrate for glucohydrolase II

Cardenolide glucohydrolase II (CGH II) is a cardenolide-specific glucohydrolase obtained from Digitalis lanata leaves. We investigated the structure-specificity relationship of several cardenolide disaccharides as a substrate for CGH II. Conformation analysis of the substrates was performed using molecular mechanics calculations. The sugar chain conformation of two inert glycosides was significantly different from that of the other glycosides. The other two glycosides, which were weak substrates of CGH II, were suggested to have an intramolecular hydrogen bond between the sugar groups. It was deduced that this hydrogen bond restricts the conformational change of the sugar chain and prevents the glycosides from enzymatic recognition.     

Ab initio MO Calculations of benzene + TCNE and naphthalene + TCNE complexes with STO-3G π-split basis set

Ab initio SCF MO calculations have been carried out on benzene + TCNE (tetracyanoethylene) and naphthalene + TCNE complexes with the STO -3G, STO -3G π-split (STO -3G for π orbitals and a split basis for π orbitals), and 4–31G basis sets. The interaction energy, gross charges, dipole moment, and the electron density in the middle plane of the complexes have also been evaluated. The STO -3G π-split basis set is appropriate for the calculation of large π–π stacking complexes from two points of view, production of reliable results and ease of computations. The approximation scheme based on the semiorthogonalized orgitals is revealed to be very efficient to save CPU time and storage in such calculations. The stable conformation and the charge-transfer interaction of the two complexes are discussed on the basis of the calculated quantities.     

Pyrimidines. XIII. Novel pyrimidine to pyrimidine transformation reactions

Novel pyrimidine to pyrimidine transformation reactions are described. 1,3-Dimethyl-(or diethyl)-uracil(1) are converted into isocytosine, 2-thiouracil or uracil derivatives by treatment with guanidine, thiourea or urea, respectively. The latter two cases require base catalysis. The effects of some substituents at C-5 and C-6 of 1,3-dialkylated uracils (1a → 1e) on this transformation were examined and a plausible mechanism is offered for their reaction. The utility of this reaction is exemplified by the facile two-step conversion of pseudouridine into the anlileukemic agent, pseudoisocytidine, in good overall yield.     

Synthetic alpha-mannosyl ceramide as a potent stimulant for an NKT cell repertoire bearing the invariant Valpha19-Jalpha26 TCR alpha chain

A NKT cell repertoire is characterized by the expression of the Valpha19-Jalpha26 invariant TCR alpha chain (Valpha19 NKT cell). This repertoire, as well as a well-established Valpha14-Jalpha281 invariant TCR alpha(+) NKT cell subset (Valpha14 NKT cell), has been suggested to have important roles in the regulation of the immune system and, thus, is a major therapeutic target. Here, we attempted to find specific antigens for Valpha19 NKT cells. Valpha19 as well as Valpha14 NKT cells exhibited reactivity to alpha-galactosyl ceramide (alpha-GalCer). Thus, a series of monoglycosyl ceramides with an axially oriented glycosidic linkage between the sugar and ceramide moiety were synthesized and their antigenicity to Valpha19 NKT cells was determined by measuring their immune responses in culture with glycolipids. Comprehensive examinations revealed substantial antigenic activity for Valpha19 NKT cells by alpha-mannosyl ceramide.     

Polymerization of acrylamide initiated with pinacol–ceric ion redox system

Polymerization of acrylamide initiated with a pinacol–ceric ion redox system was investigated. The polymer obtained was found to contain one cerium atom in a polymer molecule. It was considered that the cerium atom was introduced into the polymer molecule by the termination reaction as there is no cerium atom in the initiating radical in the present system. A similar termination reaction was attained by ferric ion but not by cerous ion. The metal ion was considered to terminate the polymerization to form a stable polymer. Some considerations on the structure of the reaction product relating to the polymerization mechanism were discussed.     

Structural and spectroscopic characterization of (mu-hydroxo or mu-oxo)(mu-peroxo)diiron(III) complexes: models for peroxo intermediates of non-heme diiron proteins

(mu-Hydroxo or oxo)(mu-1,2-peroxo)diiron(III) complexes having a tetradentate tripodal ligand (L) containing a carboxylate sidearm [Fe2(mu-OH or mu-O)(mu-O2)(L)2]n+ were synthesized as models for peroxo-intermediates of non-heme diiron proteins and characterized by various physicochemical measurements including X-ray analysis, which provide fundamental structural and spectroscopic insights into the peroxodiiron(III) complexes.     

Dynamics of order formation by colloidal adsorption onto a substrate studied with Brownian dynamics

Colloidal adsorption and spontaneous ordering of adsorbed particles on a substrate was simulated using a three-dimensional simulation model for colloidal dispersion system with an adsorptive surface under a specified bulk concentration, where the particle-particle and particle-substrate interactions were modeled on the DLVO theory. The key process for order formation is considered to be the adsorption of a particle that induces the transition from incomplete order to perfect order, and is found to involve a stochastic nature due to an energy barrier which must be overcome for the system to reach ordered state. Also, a model was developed to predict the energy barrier for order formation based on direct observation of the key process. Further, a model to describe the stochastic nature of the process was developed and its quantitative validity was demonstrated. Through the examination of the key process, it is concluded that the mechanism of the order formation is composed of two successive processes and the rate-determining step varies depending on the ionic strength.     

Carrier generation process on photoconductive polymer films as studied by magnetic field effects on the charge-transfer fluorescence and photocurrent

We have studied the magnetic field effects (MFEs) on the charge-transfer fluorescence and transient photocurrent of a 1,2,4,5-tetracyanobenzene-doped poly(N-vinylcarbazole) film, which reflect the recombination and escape yields of the carriers, respectively. The recombination yield dependence of the external magnetic field (B) clearly shows two types of the MFEs, growth with increasing B due to the hyperfine mechanism (HFM) and a negative dip due to the level-crossing mechanism (LCM). On the other hand, the escape yield indicates complementary MFEs with a sharp decrease in yield with increasing B and then a positive dip. Simultaneous observation of the HFM- and LCM-MFEs proves the stepwise hole-hopping mechanism rather the long-range hole-jumping one. The quantitative analysis of the recombination and escape MFEs is performed using the stochastic Liouville equations (SLE) for a one-dimensional lattice model in which the stepwise hole hops take place between the nearest neighbor carbazole units with spin conservation. The SLE analysis provides the recombination and hole transfer rate constants of 7.0 x 10(7) and 4.5 x 10(8) s(-1), respectively. The boundary site number for the ion pairs in the one-dimensional model is estimated by the best fit to the experimental results. The interionic distance of the boundary ion pair in the one-dimensional model including eight sites agrees with the thermalization distance in the Onsager model. Hence, it is concluded that the elementary processes in the Onsager model applied to molecular amorphous solids are the stepwise hole hops rather than a long-range hole jump.