Estimation of single-nucleotide polymorphism allele frequency by alternately binding probe competitive polymerase chain reaction

A modified solvent microextraction with back extraction method (SME/BE) combined with high performance liquid chromatography and fluorescence detection (HPLC-FD) was developed for the determination of citalopram in human plasma. Extraction process was performed in a home-made total glass vial without using a teflon ring, usually employed in SME/BE. Citalopram was first extracted from 0.5 mL of plasma, modified with sodium hydroxide, into hexane. Back extraction step was then performed into 5.2 μL of 45 mM ammonium formate solution (pH 4) using a GC microsyringe. The extract was subsequently transferred into a liner-like vial and then injected into the HPLC system. An enrichment factor of 150 along with a good sample clean-up was obtained. The calibration curve showed linearity in the range of 1.0–130.0 ng mL⁻¹ with regression coefficient corresponding to 0.992. This range covers therapeutic window and even lower amounts which is important in pharmacokinetic studies. Limits of detection and quantification, based on a signal to noise ratio (S/N) of 3 and 10, were 0.3 and 0.8 ng mL⁻¹, respectively. The method was also applied for the determination of citalopram in plasma samples after oral administration of 40 mg single dose of citalopram.     

Very strong hydrogen bonds in a bent chain structure of fluorohydrogenate anions in liquid Cs(FH)2.3F

The structure of liquid Cs(FH)(2.3)F was revealed using a combination of high-energy x-ray and neutron diffraction measurements. We found that the strongest intermolecular H-F hydrogen bonds at an average distance of 1.36 A are accompanied by the formation of a high degree of bending of the oligomer chain in the melt, with [angle]FHF=150 degrees . A reverse Monte Carlo simulation showed that the average number of atoms per chain is 4.4. A detailed chain analysis of the atomic configuration revealed that (FH)(2)F(-) oligomer chains are the major entities in the liquid, and asymmetrical FHF(-) are formed owing to the strong H-F hydrogen bonds. The results suggest that an average of one or two HF molecules bond to each of the 11 fluorine atoms surrounding a cesium ion.     

Structure-based engineering of a plant type III polyketide synthase: formation of an unnatural nonaketide naphthopyrone

Pentaketide chromone synthase (PCS) from Aloe arborescens is a novel plant-specific type III polyketide synthase (PKS) that produces 5,7-dihydroxy-2-methylchromone from five molecules of malonyl-CoA. On the basis of the crystal structures of wild-type and M207G mutant PCS, the F80A/Y82A/M207G triple mutant was constructed and shown to produce an unnatural novel nonaketide naphthopyrone by sequential condensations of nine molecules of malonyl-CoA. This is the first demonstration of the formation of a nonaketide by the structurally simple type III PKS. A homology model predicted that the active-site cavity volume of the triple mutant is increased to 4 times that of the wild-type PCS.     

Alkylation of active methylene compounds with alcohols catalyzed by an iridium complex

Base-free catalytic alpha-alkylation of active methylene compounds with primary alcohols was successfully achieved using an [IrCl(cod)](2) complex in the presence of PPh(3) to afford the corresponding saturated alpha-alkylated products in good yields.     

Time-resolved in situ ATR-IR observations of the process of sorption of water into a poly(2-methoxyethyl acrylate) film

A process of water sorption into a biocompatible polymer, poly(2-methoxyethyl acrylate) (PMEA), was investigated by time-resolved, in situ, attenuated total reflection infrared spectroscopy. Evidence for three different types of hydrated water in PMEA, that is, nonfreezing water, freezing bound water, and freezing water, were found. Each hydration structure was elucidated at the functional group level. Nonfreezing water, which never crystallizes, even at -100 degrees C, has a C=O...H-O type of hydrogen bonding interaction with the carbonyl group of PMEA. Freezing bound water, which crystallizes in a heating process below 0 degrees C, interacts with the methoxy moiety in the PMEA side chain terminal. Freezing water, which crystallizes approximately 0 degrees C, has bulk-water-like structure with an O-H...O-H hydrogen bonds network. It has been concluded from the present study that the methoxy moiety in the PMEA side chain terminal plays an important role for the excellent biocompatibility of PMEA.     

Teaching-Learning of Evaluative Criteria for Mathematical Arguments Through Classroom Discourse: A Cross-National Study

Videotaped lessons of 5th graders on equivalent fractions from 7 American and 6 Japanese classrooms were analyzed in terms of a recurrent pattern in public discourse among a teacher and students. This pattern—called inquiry, response, feedback—occurs when a teacher initiates discourse (mostly with an inquiry), a student or students respond (often with an answer to the teacher inquiry), and the teacher provides feedback to the student's response. We found2 approaches to the teaching-learning of the criteria for evaluating mathematical arguments. In the Japanese classroom, students were encouraged to offer their own argument to the whole class and evaluate arguments proposed by other students. They seldom were given direct evaluation by their teacher. In contrast, American teachers often gave individual elaboration as well as direct evaluation to the student's responses, and some of the teachers offered their own opinions about mathematics, about valid ways of argumentation, or about both. The Japanese approach would help students acquire evaluative criteria indirectly through participating in mathematical discourse, whereas the American approach would help students learn modes of arguments through direct instruction.     

Imaging of chemical reactivity and buckled dimers on Si(100)2×1 reconstructed surface with noncontact AFM

We have investigated the force interactions between the Si tip and the Si(100)2×1 reconstructed surface in the noncontact atomic-force microscopy (AFM) measurement. We observed two types of frequency shift curves without and with discontinuity, similar to the Si(111)7×7 surface. The image contrast changes drastically whether the frequency shift curve shows discontinuity or not. In the case of the frequency shift curves without discontinuity, the noncontact AFM images almost reflect the surface topography including dimers and adsorbates. In the case of the frequency shift curves with discontinuity, they reflect strongly the chemical reactivity of surface. Furthermore, in the case of the frequency shift curves without discontinuity, for the first time, the stabilize-buckling of dimers induced by a defect can be observed. This suggests that the force interactions during the noncontact AFM measurement hardly influence the surface dynamics.     

Near-field optical imaging using force detection with new tip-electrode geometry

We propose a new tip-electrode geometry to detect an (optical) evanescent field using noncontact atomic force microscopy. Using a semi-transparent metal electrode on the prism surface, the force sensitivity due to evanescent field in new tip-electrode geometry was enhanced by a factor of about 1000, comparing with that in old tip-electrode geometry where electrode was located behind the prism. Furthermore, this tip-electrode geometry avoids the electrostatic field caused by the residual charges and contact-electrified charges near the prism surface, which affects the force sensitivity due to evanescent field. We demonstrated the high resolution imaging of the evanescent field on the Au film with 15-nm (λ/33) lateral resolution.