Efficient and versatile synthesis of novel 2alpha-substituted 1alpha,25-dihydroxyvitamin D(3) analogues and their docking to vitamin D receptors.

Novel 2alpha-substituted 1alpha,25-dihydroxyvitamin D(3) analogues with 2alpha-alkyl and 2alpha-hydroxyalkyl groups were systematically synthesized from D-xylose. Their conformation on binding to the ligand binding domain (LBD) of the vitamin D receptor was analyzed. It has been found that the 2alpha-hydroxypropyl group best fits the cavity of the LBD, and the binding activity is three times higher than that for the natural hormone.     

Interaction between charged soft microcapsules and red blood cells: effects of PEGylation of microcapsule membranes upon their surface properties

Four types of hydrophilic gel microcapsules containing water have been prepared by an interfacial polymerization method. Each type of microcapsules has a membrane of different composition. Using three kinds of monomers, N,N-dimethylacrylamide (DMAAm), 4-(aminomethyl)styrene (AmSt), and N,N-dimethylaminopropylacrylamide (DMAPAA), one type of aqueous copolymer having primary and tertiary amino groups was obtained. By the polymerization of three kinds of monomers, DMAAm, AmSt, and 2-[(methacryloyloxy)ethyl] trimethylammoniumchloride (METAC), another type of aqueous copolymer having primary and quaternary ammonium groups was also obtained. Two more types of copolymers were synthesized by copolymerization of -acryloxy-ω-methoxy-poly(ethylene glycol) (a-PEG) with the above two kinds of monomer mixture. These copolymers were polymerized with terephthaloyldichloride at the water/oil interface to prepare four types of microcapsules containing water, i.e., poly(DMAAm-co-DMAPAA-co-AmSt-alt-terephthalic acid) microcapsules, poly(DMAAm-co-DMAPAA-co-AmSt-co-PEG-alt-terephthalic acid) microcapsules, poly (DMAAm-co-METAC-co-AmSt-alt-terephthalic acid) microcapsules, and poly (DMAAm-co-METAC-co-AmSt-co-PEG-alt-terephthalic acid) microcapsules, which will be abbreviated to MC 1, MC 2, MC 3, and MC 4, respectively. It has been predicted that the microcapsule membranes are hydrophilic and soft and have two-sublayer structures from electrophoretic mobility measurements and from the analysis of the data with Ohshima’s electrokinetic theory for soft particles. The outer sublayers of MC 1 and MC 2 are negatively charged and those of MC 3 and 4 are slightly positively charged. Also, the surfaces of MC 1 and MC 2 are harder than those of MC 3 and 4. By PEGylation, the surface charge density in the membranes decreases and the surface becomes softer. It has been found that the membrane of red blood cells (RBC) is also soft and is composed of two-sublayers, the outer sublayer of which is negatively charged and the inner one is positively charged. The interaction of four types of microcapsules with RBC has been studied. It was found that microcapsules with soft surfaces (MC 3 and MC 4) do not interact with RBC, even though the microcapsule surfaces are positively charged and the surface of RBC is negatively charged. On the other hand, microcapsules with negatively charged but harder surfaces (MC 1) interact with RBC to introduce hemolysis. The membrane surface of MC 2, which is obtained by PEGylation of MC 1, becomes softer than that of MC 1 so that the interaction with RBC was weakly suppressed. From these, it was concluded that the dominant factor to control the interaction between synthetic polymer surfaces and biological cell surfaces is not the surface charges carried by the polymer surfaces but the softness of the polymer surfaces.     

Synthesis and crystallization-induced microphase separation of cellulose triacetate-block-poly(γ-benzyl-l-glutamate)

This article describes the first observation of crystallization-induced microphase separation in thin film and bulk cellulose triacetate-block-poly(γ-benzyl-l-glutamate) (PBLG) [cellulose triacetate (CTA)-b-PBLG] via copper-catalyzed azide–alkyne cycloaddition (CuAAC) between azido-functionalized CTA at the reducing end and alkyne-functionalized PBLG at the C-terminus. The reactivity of the amino group at the C-1 position of the glucosyl residue at the reducing end for the initiation reaction of the ring-opening polymerization (ROP) of γ-benzyl-l-glutamate N-carboxyanhydride was compared to that of the azido group at the reducing end of CTA for CuAAC, with PBLG bearing an alkyne group at the C-terminus. Although the amino group at the reducing end of CTA exhibited no reactivity as a macroinitiator for ROP of BLG, the azido group at the reducing end of CTA reacted with the alkyne group at the C-terminus of PBLG to afford CTA-b-PBLG. The structure of CTA-b-PBLG was characterized by ¹H- and ¹³C-nuclear magnetic resonance spectroscopies, infrared spectroscopy, differential scanning calorimetry, and wide angle X-ray diffractometry. Microphase separation of the film and bulk of CTA-b-PBLG was clearly shown by atomic force microscopy, field-emission scanning electron microscopy, and transmission electron microscopy.     

Electrochemical decomposition of layer-by-layer thin films composed of TEMPO-modified poly(acrylic acid) and poly(ethyleneimine)

The decomposition of layer-by-layer (LbL) thin films composed of 2,2,6,6-tetramethylpiperidine-1-oxyl free radical-appended poly(acrylic acid) (TEMPO-PAA) and poly(ethylenimine) (PEI) was studied by using a quartz crystal microbalance (QCM) and cyclic voltammetry. The electrode potential of the (PEI/TEMPO-PAA)₄/PEI film-coated Au resonator was scanned from +0.2 to +0.8 V vs Ag/AgCl. The CV showed that the oxidation peak current decreased as the number of scans increased. The change in the resonance frequency of the QCM increased after electrolysis, indicating that the film was decomposed by electrolysis. The positive charges originating from the oxoammonium ions probably destabilized the (PEI/TEMPO-PAA)₄/PEI film. Furthermore, the release of 5,10,15,20-tetraphenyl-21H,23H-porphine tetrasulfonic acid (TPPS) from TPPS-loaded (PEI/TEMPO-PAA)₄/PEI-coated ITO electrodes was investigated. TPPS was released at electrode potentials greater than +0.6 V by the decomposition of the film. The results suggest that TEMPO-PAA/PEI LbL films are suitable for electrochemically controlled drug delivery systems.     

Conformation-dependent ligand regulation of ATP hydrolysis by human KSP: activation of basal hydrolysis and inhibition of microtubule-stimulated hydrolysis by a single, small molecule modulator

Human kinesin spindle protein (KSP)/hsEg5, a member of the kinesin-5 family, is essential for mitotic spindle assembly in dividing human cells and is required for cell cycle progression through mitosis. Inhibition of the ATPase activity of KSP leads to cell cycle arrest during mitosis and subsequent cell death. Ispinesib (SB-715992), a potent and selective inhibitor of KSP, is currently in phase II clinical trials for the treatment of multiple tumor types. Mutations that attenuate Ispinesib binding to KSP in vitro have been identified, highlighting the need for inhibitors that target different binding sites and inhibit KSP activity by novel mechanisms. We report here a small-molecule modulator, KSPA-1, that activates KSP-catalyzed ATP hydrolysis in the absence of microtubules yet inhibits microtubule-stimulated ATP hydrolysis by KSP. KSPA-1 inhibits cell proliferation and induces monopolar-spindle formation in tumor cells. Results from kinetic analyses, microtubule (MT) binding competition assays, and hydrogen/deuterium-exchange studies show that KSPA-1 does not compete directly for microtubule binding. Rather, this compound acts by driving a conformational change in the KSP motor domain and disrupts productive ATP turnover stimulated by MT. These findings provide a novel mechanism for targeting KSP and perhaps other mitotic kinesins.     

Analysis of incommensurate structure in [N(CH3)4]2ZnCL4 crystal

We analyzed the temperature dependence of the amplitude of modulated structures of [N(CH₃)₄]₂ZnCl₄ crystal in the incommensurate (INC) phase by measuring the EPR spectra for Mn²⁺ doped single crystal. The amplitude of the vibrational modulation linearly increased with decreasing temperature. Applying Frank and Van der Merwe model, we simulated the EPR spectra in the INC phase. The simulated spectra agreed with the observed one very well. We, therefore, revealed that the vibrational modulation in this INC phase is due to the interaction between the harmonic chain of inter-particles and the modulation due to underlying potential which comes from the commensurate structure.     

2α + t Cluster Structure in 11B

Structures of excited states in ¹¹B are investigated with a method of β – γ constraint antisymmetrized molecular dynamics in combination with the generator coordinate method. Various cluster structures are suggested in excited states. For negative-parity states, we suggest a band with a 2α + t cluster structure. This band starts from the ${3/2^{-}_{3}}$ state and can correspond to the experimental band observed recently. We find that the feature of the ${3/2^{-}_{3}}$ is quite similar to the ${0^{+}_{2}}$ state in ¹²C.     

Chip-based frequency combs with sub-100 GHz repetition rates

By fabricating high-Q silicon-nitride spiral resonators, we demonstrate frequency combs spanning over 200 nm with free spectral ranges (FSRs) of 80, 40, and 20 GHz using cascaded four-wave mixing. We characterize the RF beat note for the 20 GHz FSR comb, and the measured linewidth of 3.6 MHz is consistent with thermal fluctuations in the resonator due to amplitude noise of the pump source. These combs represent an important advance towards developing a complementary metal-oxide-semiconductor (CMOS)-based system capable of linking the optical and electronic regimes.     

Development of radioligands for the dopamine transporter

The extraction of Penicillin G (Pen G) and its conversion to 6-aminopenicillin acid (6-APA) and phenylacetic acid (PAA) was performed by means of Penicillin G Amidase immobilised in the emulsion liquid membranes. Using various surfactants as emulsifiers and an appropriate carrier it is possible to obtain different extraction rates of Pen G as well as back transfer rates of the hydrolysis products. The surfactants with polyoxyethylene chain facilitate the back transfer through the membrane phase, whereas the more hydrophobic surfactants, e.g., Paranox 100, tend to accumulate Pen G hydrolysis products in the internal aqueous phase.