Modeling of extraction behavior of docosahexaenoic acid ethyl ester by utilizing slug flow prepared by microreactor

The liquid–liquid extraction dynamics of an ethyl ester of docosahexaenoic acid (DHA‐Et) with silver ion was investigated. The kinetic model was derived according to the following stepwise processes: Diffusion of DHA‐Et across the organic film, complex‐formation between DHA‐Et and silver ion at the interface, and diffusion of extracted complex across the aqueous film. The kinetic parameters for the complex‐formation reaction were determined from the investigation with the stirred transfer cell. With the proposed model and determined parameters, we predicted the uptakes of DHA‐Et for the extraction system utilizing a slug flow prepared by a microchip. The calculated uptakes showed good correlation to the experimental data. The theoretical investigation suggested that the fast equilibration realized for the slug flow extraction system was due to the large specific interfacial area of the slug caused by the presence of wall film and the thin liquid film caused by the internal circulation. © 2009 American Institute of Chemical Engineers AIChE J, 2010     

Analyses of liquid film models applied to horizontal and near horizontal gas–liquid slug flows

An analysis of liquid film models for horizontal and near horizontal gas–liquid slug flows is developed. The models’ formulations employ the one dimensional separated phase momentum equations. The formulations differ among themselves, by neglecting some terms on the momentum balance and also on the closure relations. A comparative analysis discloses the differences amongst the formulations. The sensitivity of the liquid film models to the changes on the bubble velocity, liquid slug holdup and liquid viscosity is accessed through a series of parametric runs. Finally, the model is tested against experimental data taken for continuous horizontal slug flow. The tests were designed to check if the models are able to capture the stochastic film properties provided the properly closure relations.     

Effect of Ganglioside GM3 Synthase Gene Knockout on the Glycoprotein N‐Glycan Profile of Mouse Embryonic Fibroblast

The structural and clinical significance of cellular glycoproteins and glycosphingolipids (GSLs) are often separately discussed. Considering the biosynthetic pathway of glycoconjugates, glycans of cell‐surface glycoproteins and GSLs might partially share functions in maintaining cellular homeostatis. The purpose of this study is to establish a general and comprehensive glycomics protocol for cellular GSLs and N‐glycans of glycoproteins. To test the feasibility of a glycoblotting‐based protocol, whole glycans released both from GSLs and glycoproteins were profiled concurrently by using GM3 synthase‐deficient mouse embryonic fibroblast GM3(?/?). GM3(?/?) cells did not synthesize GM3 or any downstream product of GM3 synthase. Instead, expression levels of o‐series gangliosides involving GM1‐b and GD1‐α increased dramatically, whereas a‐/b‐series gangliosides were predominantly detected in wild‐type (WT) cells. We also discovered that glycoprotein N‐glycan profiles of GM3(?/?) cells are significantly altered as compared to WT cells, although GM3 synthase is responsible only for GSLs synthesis and is not associated with glycoprotein N‐glycan biosynthesis. The present approach allows for high‐throughput profiling of cellular glycomes enriched by different classes of glycoconjugates, and our results demonstrated that gene knockout of the enzymes responsible for GSL biosynthesis significantly influences the N‐glycans of glycoproteins.     

A Single Mutation at the Ferredoxin Binding Site of P450 Vdh Enables Efficient Biocatalytic Production of 25‐Hydroxyvitamin D3

Vitamin D₃ hydroxylase (Vdh) from Pseudonocardia autotrophica is a cytochrome P450 monooxygenase that catalyzes the two‐step hydroxylation of vitamin D₃ (VD₃) to produce 25‐hydroxyvitamin D₃ (25(OH)VD₃) and 1α,25‐dihydroxyvitamin D₃ (1α,25(OH)₂VD₃). These hydroxylated forms of VD₃ are useful as pharmaceuticals for the treatment of conditions associated with VD₃ deficiency and VD₃ metabolic disorder. Herein, we describe the creation of a highly active T107A mutant of Vdh by engineering the putative ferredoxin‐binding site. Crystallographic and kinetic analyses indicate that the T107A mutation results in conformational change from an open to a closed state, thereby increasing the binding affinity with ferredoxin. We also report the efficient biocatalytic synthesis of 25(OH)VD₃, a promising intermediate for the synthesis of various hydroxylated VD₃ derivatives, by using nisin‐treated Rhodococcus erythropolis cells containing Vdh_T107A. The gene‐expression cassette encoding Bacillus megaterium glucose dehydrogenase‐IV was inserted into the R. erythropolis chromosome and expressed to avoid exhaustion of NADH in a cytoplasm during bioconversion. As a result, approximately 573 μg mL^?1 25(OH)VD₃ was successfully produced by a 2 h bioconversion.     

Round robin on indentation fracture resistance of silicon carbide ceramics by using a powerful optical microscope

Reproducibility of indentation fracture resistance, K_IFR of silicon carbides sintered with B and C was evaluated by a round robin with ten laboratories. When the crack length was measured with an optical microscope at a low magnification of ～100×, K_IFR varied widely from 3.43 to 4.20 MPa m^1/2, whereas those obtained by a powerful microscopy with both an objective lens of 40× and a traveling stage exhibited a consistent value of 3.20±0.12 MPa m^1/2. The wide scatter of K_IFR for the former measurements was attributed mainly to the variation in misreading of the crack length. It was revealed that the high resolving power of the objective lens of 40× enabled to find exact crack tips easily, which resulted in the good matching of K_IFR between laboratories for the latter case. It was suggested that the observation of indentations with powerful optics was effective for improving the reproducibility of the IF method.     

Allelopathic Effects of Volatile Monoterpenoids Produced by <Emphasis Type="Italic">Salvia leucophylla</Emphasis>: Inhibition of Cell Proliferation and DNA Synthesis in the Root Apical Meristem of <Emphasis Type="Italic">Brassica campestris</Emphasis> Seedlings

Salvia leucophylla, a shrub observed in coastal south California, produces several volatile monoterpenoids (camphor, 1,8-cineole, -pinene, -pinene, and camphene) that potentially act as allelochemicals. The effects of these were examined using Brassica campestris as the test plant. Camphor, 1,8-cineole, and -pinene inhibited germination of B. campestris seeds at high concentrations, whereas -pinene and camphene did not. Root growth was inhibited by all five monoterpenoids in a dose-dependent manner, but hypocotyl growth was largely unaffected. The monoterpenoids did not alter the sizes of matured cells in either hypocotyls or roots, indicating that cell expansion is relatively insensitive to these compounds. They did not decrease the mitotic index in the shoot apical region, but specifically lowered mitotic index in the root apical meristem. Moreover, morphological and biochemical analyses on the incorporation of 5-bromo-2-deoxyuridine into DNA demonstrated that the monoterpenoids inhibit both cell-nuclear and organelle DNA synthesis in the root apical meristem. These results suggest that the monoterpenoids produced by S. leucophylla could interfere with the growth of other plants in its vicinity through inhibition of cell proliferation in the root apical meristem.     

Inference of gene regulation via miRNAs during ES cell differentiation using MiRaGE method

MicroRNA (miRNA) is a critical regulator of cell growth, differentiation, and development. To identify important miRNAs in a biological process, many bioinformatical tools have been developed. We have developed MiRaGE (MiRNA Ranking by Gene Expression) method to infer the regulation of gene expression by miRNAs from changes of gene expression profiles. The method does not require precedent array normalization. We applied the method to elucidate possibly important miRNAs during embryonic stem (ES) cell differentiation to neuronal cells and we infer that certain miRNAs, including miR-200 family, miR-429, miR-302 family, and miR-17-92 cluster members may be important to the maintenance of undifferentiated status in ES cells.     

Fabrication of fluorine-terminated diamond-like carbon thin film using a hyperthermal atomic fluorine beam

Surface modification of diamond-like carbon (DLC) film was performed using a hyperthermal atomic fluorine beam on the purpose of production of hydrophobic surface by maintaining the high hardness of DLC film. By the irradiation of atomic fluorine beam of a 1.0 × 10²⁰ atoms/cm², the contact angle of a water drop against the DLC surface increased from 73° to 111°. The formation of CF₃, CF₂ and CF bonding on the modified DLC surface was confirmed from the measurements of X-ray photoelectron spectra and near-edge X-ray absorption fine structure spectra. Irradiation of hyperthermal atomic fluorine beam was concluded to produce insulator fluorine-terminated DLC film, which has high F content on the surface, by the taking of the use of neutral atomic beam as a fluorine source.     

Combustion synthesis of LaSi3N5:Eu2+ phosphor powders

LaSi₃N₅:Eu²⁺ phosphor powders were prepared by a highly efficient combustion synthesis method. It was found that the compositions of the raw powder mixtures had great influences on the phase compositions and particle morphologies of the synthesized powders. By selecting appropriate starting compositions and combustion parameters, single phase LaSi₃N₅:Eu²⁺ phosphors could be synthesized. When excited by a UV light, the LaSi₃N₅:Eu²⁺ phosphors emitted green light. The wavelength and intensity of the emission spectra were affected by the amount of Eu²⁺ dopant. With increasing amount of Eu²⁺ dopant, concentration quenching could occur and emission spectra shifted to longer wavelengths.     

A New Lead Identification Strategy: Screening an sp3-rich and Lead-like Compound Library Composed of 7-Azanorbornane Derivatives

Although the advantages of sp³-rich, sterically complicated molecules in drug development have been pointed out, modern screening libraries are filled with planar, sp²-rich components. Compounds that are sp³-rich are difficult to synthesize, and thus we aimed to invent an efficient method to construct sp³-rich libraries. By modifying sp³-rich 7-azanorbornane scaffolds through click chemistry, we efficiently prepared a small set of compounds. These compounds were not only sp³-rich, but also had sufficient “lead-like” properties in view of molecular weights and hydrophobicity. Screening assays of this library provided weak κ opioid receptor agonists and growth hormone secretagogue receptor agonists with high hit rates. These results indicate that the 7-azanorbornane scaffold may be a “privileged structure” for lead identification in drug discovery.