Application of dual counter-current chromatography for rapid sample preparation of N-methylcarbamate pesticides in vegetable oil and citrus fruit

Dual counter-current chromatography (dual CCC) has been successfully applied to rapid sample preparation for the simultaneous determination of residual carbaryl, fenobucarb and methomyl in vegetable oil and citrus fruit. The citrus fruit samples were extracted with n-hexane solution containing stable isotopically labeled internal standards (methomyl-d3, fenobucarb-d3 and carbaryl-d9), and applied to dual CCC using a two-phase solvent system of n-hexane-acetonitrile to purify the carbamate pesticides from aliphatic sample matrix. The coiled column was rotated at 420 rpm, the lower mobile phase was introduced through the head toward the tail, and the upper mobile phase in the opposite direction. Due to the high partition efficiency of dual CCC, the lower phase fraction collected from 2 to 5 min after injection could be subjected to flow-injection tandem mass spectrometry directly after concentration. Repetitive sample injection can be performed at high reproducibility without a risk of contamination from the compounds retained in the column.     

Synthesis, DNA polymerase incorporation, and enzymatic phosphate hydrolysis of formamidopyrimidine nucleoside triphosphates

The nucleoside triphosphates of N6-(2-deoxy-alpha,beta-d-erythro-pentofuranosyl)-2,6-diamino-4-hydroxy-5-formamidopyrimidine (Fapy.dGTP) and its C-nucleoside analogue (beta-C-Fapy.dGTP) were synthesized. The lability of the formamide group required that nucleoside triphosphate formation be carried out using an umpolung strategy in which pyrophosphate was activated toward nucleophilic attack. The Klenow fragment of DNA polymerase I from Escherichia coli accepted Fapy.dGTP and beta-C-Fapy.dGTP as substrates much less efficiently than it did dGTP. Subsequent extension of a primer containing either modified nucleotide was less affected compared to when the native nucleotide is present at the 3'-terminus. The specificity constants are sufficiently large that nucleoside triphosphate incorporation could account for the level of Fapy.dG observed in cells if 1% of the dGTP pool is converted to Fapy.dGTP. Similarly, polymerase-mediated introduction of beta-C-Fapy.dG could be useful for incorporating useful amounts of this nonhydrolyzable analogue for use as an inhibitor of base excision repair. The kinetic viability of these processes is enhanced by inefficient hydrolysis of Fapy.dGTP and beta-C-Fapy.dGTP by MutT, the E. coli enzyme that releases pyrophosphate and the corresponding nucleoside monophosphate upon reaction with structurally related nucleoside triphosphates.     

Increased expression and phosphorylation of liver glutamine synthetase in well-differentiated hepatocellular carcinoma tissues from patients infected with hepatitis C virus

Hepatocellular carcinoma (HCC) is one of the most common fatal cancers, and chronic infection with hepatitis C virus (HCV) is thought to be one of the main causes in Japan. To identify diagnostic or therapeutic biomarkers for HCC associated with HCV (HCV-HCC), we tried to elucidate the factors related to the products from cancerous tissues of HCV-infected patients. From proteomic differential display analysis of liver tissue samples from HCV-HCC cancerous tissues and corresponding non-cancerous tissues from patients, three protein spots of the same molecular mass (42 kDa), whose expression increased in well-differentiated cancerous tissues, were detected. Although their pI were different, they were identified as glutamine synthetase (GS) by PMF with MALDI-TOF MS and by Western blotting using anti-GS specific mAb. Immunohistochemical analysis showed that tumor tissue consists of two parts, GS-positive cell and GS-negative cell regions, suggesting that GS-producing cells grew in the tumor tissue as a nodule in nodules. The tryptic peptides of the most acidic GS isoform lost the signal of 899.5 Da, corresponding a peptide of SASIRIPR, and gained a signal of 1059.5 Da, which was submitted to PSD analysis. PSD analysis showed the neutral loss by elimination of two phosphate groups, supposed to be on serine residues of the 899.5-Da peptide, from serine 320 to arginine 327 in GS. PMF followed by PSD analysis is thought to be useful for the determination of phosphorylation sites of proteins showing molecular heterogeneity.     

High birefringence of the yttrium vanadate crystal in the middle wavelength infrared

A high birefringence of over 0.21 for the yttrium vanadate (YVO4) crystal in the middle wavelength infrared (i.e., 3-5 microm) was measured. A Fourier transform infrared spectrometer was employed in the channel spectra technique to obtain the measurements.     

Stabilization of a channeled spectropolarimeter by self-calibration

A novel method to stabilize a channeled spectropolarimeter is described. The fluctuating retardations of the high-order retarders used in the spectropolarimeter are calibrated in parallel to the measurement of the wavenumber-dependent state of polarization (SOP) of light. Both the calibration of the retarders and the measurement of the SOP can be made simultaneously using a single light to be measured, and hence the resultant wavenumber-dependent SOP is almost immune to fluctuation of the retardations. The effectiveness of this method is experimentally demonstrated with retardation fluctuations induced by a temperature change up to 40 degrees C.     

Structural phase transition,antiferromagnetism and two superconducting domes in LaFeAsO_(1-x)F_x(0<x≤0.75)

We report~(75) As nuclear magnetic resonance(NMR)/nuclear quadrupole resonance(NQR) and transmission electron microscopy(TEM) studies on LaFeAsO_(1-x)F_x. There are two superconducting domes in this material. The first one appears at 0.03 ≤ x ≤0.2 with T_c~(max) = 27 K, and the second one at 0.25 ≤x≤0.75 with T_c~(max) = 30 K. By NMR and TEM, we demonstrate that a C4-to-C2 structural phase transition(SPT) takes place above both domes, with the transition temperature T_s varying strongly with x. In the first dome, the SPT is followed by an antiferromagnetic(AF) transition, but neither AF order nor low-energy spin fluctuations are found in the second dome. By ~(75) As nuclear spin-lattice relaxation rate(1/T_1) measurements, we find that AF order and superconductivity coexist microscopically in LaFeAsO_(0.97) F_(0.03). In the coexisting region, 1/T_1 decreases at T_c but becomes proportional to T below 0.6 T_c, indicating gapless excitations. Therefore, in contrast to the early reports, the obtained phase diagram for x ≤ 0.2 is quite similar to the doped BaFe_2As_2 system. The electrical resistivity p in the second dome can be fitted by ρ = ρ0 + AT~n with n = 1 and a maximal coefficient A at around xopt = 0.5-0.55 at which T_s extrapolates to zero and Tc is the maximal, which suggests the importance of quantum critical fluctuations associated with the SPT. We have constructed a complete phase diagram of LaFeAsO_(1-x)F_x, which provides insight into the relationship between SPT, antiferromagnetism and superconductivity.     

Tissue-Specific Regulation of HNK-1 Biosynthesis by Bisecting GlcNAc

Human natural killer—1 (HNK-1) is a sulfated glyco-epitope regulating cell adhesion and synaptic functions. HNK-1 and its non-sulfated forms, which are specifically expressed in the brain and the kidney, respectively, are distinctly biosynthesized by two homologous glycosyltransferases: GlcAT-P in the brain and GlcAT-S in the kidney. However, it is largely unclear how the activity of these isozymes is regulated in vivo. We recently found that bisecting GlcNAc, a branching sugar in N-glycan, suppresses both GlcAT-P activity and HNK-1 expression in the brain. Here, we observed that the expression of non-sulfated HNK-1 in the kidney is unexpectedly unaltered in mutant mice lacking bisecting GlcNAc. This suggests that the biosynthesis of HNK-1 in the brain and the kidney are differentially regulated by bisecting GlcNAc. Mechanistically, in vitro activity assays demonstrated that bisecting GlcNAc inhibits the activity of GlcAT-P but not that of GlcAT-S. Furthermore, molecular dynamics simulation showed that GlcAT-P binds poorly to bisected N-glycan substrates, whereas GlcAT-S binds similarly to bisected and non-bisected N-glycans. These findings revealed the difference of the highly homologous isozymes for HNK-1 synthesis, highlighting the novel mechanism of the tissue-specific regulation of HNK-1 synthesis by bisecting GlcNAc.     

Mechanism of Oxygen Adsorption on Partially K Exchanged Na-A Type Zeolite

Accordingly, the observation of oxygen selectivity on Na-K-A would indicate that nitrogen has a greater steric hindrance at the window than does oxygen. Using single crystal X-ray diffraction (SCXD), it was confirmed that K at 8-member rings was relocated to 6-member rings and that the location of Na at 8-member rings became uncertain with calcination. It was also observed that Na-K-A was thermally more stable than Na-A. Using magic angle spin nuclear magnetic resonance (MAS-NMR), it was confirmed that Na at 8-member rings irreversibly changed location from a symmetric position to an asymmetric position with calcination.Using the atomic positions determined by SCXD, the mean square displacements (MSD) of oxygen and nitrogen inside the crystal of Na-A and Na-K-A were estimated by means of molecular dynamic simulation (MD). When temperature was decreased, the MSD of nitrogen was reduced to a greater extent than that of oxygen and the MSD of nitrogen inside Na-K-A was reduced by a greater margin than that inside Na-A.     

Irradiation effects on hydrases for biomedical applications

To apply an irradiation technique to sterilize “Hybrid” biomedical materials including enzymes, we selected papain, a well-characterized plant endopeptidase as a model to examine durability of enzyme activity under the practical irradiation condition in which limited data were available for irradiation inactivation of enzymes. Dry powder and frozen aqueous solution of papain showed significant durability against ⁶⁰Co-gamma irradiation suggesting that, the commercial irradiation sterilizing method is applicable without modification. Although irradiation of unfrozen aqueous papain solution showed an unusual change of the enzymatic activity with the increasing doses, and was totally inactivated at 15 kGy, we managed to keep the residual activity more than 50% of initial activity after 30-kGy irradiation, taking such optimum conditions as increasing enzyme concentration from 10 to 100 mg/ml and purging with N₂ gas to suppress the formation of free radicals.     

The dynamics of open quantum systems: accessibility results

In this paper we study the accessibility properties of finite dimensional (N-level) open quantum systems in the presence of dissipation and relaxation described by the Lindblad master equation. We specifically focus on the unital Lindbladian case where general results can be obtained. The theory of transitive Lie-group actions is used to classify the system Lie-algebras of the Lindblad equation for which the reachable sets have nonempty interior. For the special case of n -coupled spin-1/2 systems, we obtain a particularly simple characterization. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)