In situ immunocytochemical detection of altered membrane composition induced by cell-cell contact in cultured mammalian cells.

In cultured mammalian cells, both normal and transformed, cell-cell contact was shown to alter the detergent extractability of nuclear and plasma membranes detected using immunocytochemistry of fixed cells. This alteration occurred in each cell individually, occurred in less than 1 hour after altered cell-cell contact, and did not involve new protein or mRNA synthesis. These results indicate that composition of cellular membranes is highly affected by cell-cell contact and culture density.

Ferulates, amurenlactone A and amurenamide A from traditional Chinese medicine cortex Phellodendri Amurensis.

Two new ferulate compounds, amurenlactone A (1) and amurenamide A (2), along with 11 known compounds have been isolated from the cortex of Phellodendron amurense. The structures of the new compounds were established based on 1D, 2D NMR and mass spectral analyzes. The known compounds were identified by comparison with authentic samples.

A new thiaziedione from the fruits of Xanthium sibiricum.

A novel sulphur-containing compound named xanthiazinone (1) was isolated from the ethanolic extract of the fruits of Xanthium sibiricum, along with four known compounds, xanthiazone (2), xanthiside (3), xanthienopyran (4) and 5-hydroxypyrrolidin-2-one (5). Their structures were determined on the basis of spectroscopic techniques.

Real-time gray-scale photolithography for fabrication of continuous microstructure.

A novel real-time gray-scale photolithography technique for the fabrication of continuous microstructures that uses a LCD panel as a real-time gray-scale mask is presented. The principle of design of the technique is explained, and computer simulation results based on partially coherent imaging theory are given for the patterning of a microlens array and a zigzag grating. An experiment is set up, and a microlens array and a zigzag grating on panchromatic silver halide sensitized gelatin with trypsinase etching are obtained.

A high throughput perfusion-based microbioreactor platform integrated with pneumatic micropumps for three-dimensional cell culture.

This study reports a new perfusion-based, micro three-dimensional (3-D) cell culture platform for high-throughput cell culture using enabling microfluidic technologies. In this work, the micro 3-D cell culture platform is fabricated based on SU-8 lithography and polydimethylsiloxane replication processes. The micro cell culture platform can maintain homogenous and stable culture environments, as well as provide pumping of multiple mediums and efficient cell/agarose (scaffold) loading functions, which allows realization of more precise and high-throughput cell culture-based assays.

Nanoparticulate carrier containing water-insoluble iodinated oil as a multifunctional contrast agent for computed tomography imaging.

Contrast-enhanced computed tomography (CT) imaging is a valuable and routine strategy for the clinical diagnosis of various diseases. However, all current CT contrast agents are liquids, so they flow through the blood vessels and disappear very quickly by extravasation. If it were possible to make a blood-compatible particulate contrast agent, we could highlight a particular tissue by either passive or active targeting.

Unique glycerophospholipid signature in retinal stem cells correlates with enzymatic functions of diverse long-chain acyl-CoA synthetases.

Lipidomics is an emerging research field that comprehensively characterizes lipid molecular species and their metabolic regulation and biological roles. We performed the first lipidomics study on glycerophospholipids (GPLs) in adult mammalian retinal stem cells (RSCs) and non-RSC control cells. A unique GPL signature identified by electrospray ionization tandem mass spectrometry showed new prominent peaks of 16:0 (sn-1)-18:0 (sn-2) or 16:0-16:0 saturated fatty acids, instead of 18:0-20:4 or 18:0-22:6 polyunsaturated essential fatty acids, at 720 m/z of phosphatidylethanolamine, 764 m/z of phosphatidylserine, and 809 m/z of phosphatidylinositol in RSCs (sphere colony RSCs and enriched RSCs), but not in non-RSCs (retinal cells, ciliary cells, sphere colony-derived retinal cells, and nonretinal cells).

A rapid method for isolation and purification of an anticoagulant from Whitmania pigra.

Whitmania pigra is common in China and has been used as a traditional Chinese anticoagulant medicine for years, but its effective components are unknown to scientists. In this article we report a rapid method for isolation and purification of an anticoagulant from W. pigra for the first time.

Molecular cloning and characterization of a cDNA encoding a novel antibacterial peptide, defensin, from the mulberry longicorn beetle, Apriona germari.

A full-length cDNA clone with high homology (62% mature peptide sequence identity) to an Acalolepta luxuriosa antibacterial gene, possessing a conserved cysteine-stabilized alphabeta motif, was cloned by screening an Apriona germari cDNA library. This gene (AgCRP) had a total length of 360 bp with an open reading frame of 207 bp, and encoded a predicted peptide of 69 amino acid residues. The mature AgCRP peptide was 27 amino acid residues long and had a cysteine-stabilized alphabeta motif of C.

Effect of extracellular ph on matrix synthesis by chondrocytes in 3D agarose gel.

In cartilage tissue engineering, the determination of the most appropriate cell/tissue culture conditions to maximize extracellular matrix synthesis is of major importance. The extracellular pH plays an important role in affecting energy metabolism and matrix synthesis by chondrocytes. In this study, chondrocytes were isolated from bovine articular cartilage, embedded in agarose gel, and cultured at varied pH levels (7.

Study of on-line monitoring of lactate based on optical fibre sensor and in-channel mixing mechanism.

A PDMS-based microfluidic device with integrated optical fibres was developed for online monitoring of lactate. The detection is based on the optical adsorption of the colour solution formed by in-channel mixing of sample and reagent solutions. Computational Fluid Dynamics (CFD) simulation and kinetic study of the colour development were conducted to determine the minimum channel length and optimum residential time for homogenous mixing and maximum optical adsorption respectively.

Mouse orthotopic lung cancer model induced by PC14PE6.

Purpose: This study was undertaken to investigate in detail the xenograft mouse orthotopic lung cancer model induced by PC14PE6 adenocarcinoma cells.

Materials And Methods: Three cell doses (0.5x10(6); 1x10(6); 2x10(6)) of PC14PE6 cells were injected into the lungs of male BALB/c nude mice by the intrathoracic injection method.

Effector mechanisms of the anti-cancer immune responses of macrophages in SR/CR mice.

SR/CR (spontaneous regression/complete resistance) mice resist multiple types of cancer cells injected at numbers that are lethal to wild type (WT) mice. When the anti-tumor response was examined, leukocytes of the innate immune system, including neutrophils (PMN), macrophages and NK cells, infiltrated the tumor site for a multipronged killing response. Each cell type had independent killing activity against the cancer cells.

Mass spectrometry identification of circulating alpha-1-B glycoprotein, increased in aged female C57BL/6 mice.

In this study, we surveyed the profiles of mouse circulating proteins by 2-dimensional SDS-PAGE in different strains, sexes and ages. Among visible protein spots on 2-D gels with silver-staining, we identified a unique set of 7 seemingly-related proteins whose levels were consistently elevated in older C57BL/6 female mice. This set of 7 proteins was absent in C57BL/6 males or in BALB/c mice of either sex of any age.

Development of PDMS microbioreactor with well-defined and homogenous culture environment for chondrocyte 3-D culture.

Perfusion cell culture is believed to provide a stable culture environment due to the continuous supply of nutrients and removal of waste. However, the culture scales used in most cases were large, where the culture conditions can not be regarded as homogenous because of chemical gradients. To improve this, the concept of miniaturization is applied to 3-D cell culture.

Unique molecular signatures of glycerophospholipid species in different rat tissues analyzed by tandem mass spectrometry.

Glycerophospholipids (GPL) in animal tissues are composed of a large array of molecular species that mainly differ in the fatty acyl composition. In order to further understand the roles of GPL at the molecular level, it is necessary to have comprehensive, accurate accounts of the molecular makeup for these molecules in animal tissues. However, this task was difficult simply because the conventional technologies of profiling GPL species depended heavily on technical skill for accuracy and reliability and were extremely labor-intensive.

New triterpenoids isolated from the root bark of Ulmus pumila L.

Three new triterpenoids, 1-3, were isolated from the dried root bark of Ulmus pumila. Along with the three new compounds, six known triterpenoids, epifriedelanol (4), friedelin (5), oleanolic acid (6), maslinic acid (7), camaldulenic acid (8), and arjunolic acid (9) were also isolated. The structures of new compounds were established as ulmudiol (bauer-7-ene-1alpha,3beta-diol, 1), dehydroulmudiol [bauer-7,9(11)-diene-1alpha,3beta-diol, 2], and ulmuestone [3alpha-hydroxy-11alpha-(4'-hydroxy-3'-methoxy)benzoyloxybauer-1-one, 3], on the basis of extensive 1D and 2D NMR spectroscopic data interpretation.

New diterpenoids from Semiaquilegia adoxoides.

Two new ent-kaurane-type diterpenoids, E-semiaquilegin (1) and Z-semiaquilegin (2), together with eight known compounds (3-10) were isolated from the dried roots of Semiaquilegia adoxoides (DC.) Makino. The structures of compounds 1 and 2 were elucidated mainly by 2D NMR techniques including 1H-1H COSY, HSQC, HMBC, NOESY as 16alpha-hydroxy-ent-kaurane-17,20-di-(3,4-dihydroxy-E-cinnamoyl) ester and its (Z)-isomer.

Transferable anticancer innate immunity in spontaneous regression/complete resistance mice.

Spontaneous regression/complete resistance (SR/CR) mice resist very high doses of cancer cells that are lethal to WT mice even at low doses. In this study, we show that this resistance is mediated by rapid infiltration of leukocytes, mostly of innate immunity, in both primary and repeated challenges. Formation of rosettes with infiltrating natural killer cells, neutrophils, and macrophages was required for the subsequent destruction of cancer cells through rapid cytolysis.

Attenuated secretion of very low density lipoproteins from McA-RH7777 cells treated with eicosapentaenoic acid is associated with impaired utilization of triacylglycerol synthesized via phospholipid remodeling.

In McA-RH7777 cells stably expressing human apolipoprotein (apo) B100, treatment with oleic acid (18:1(n-9)) promoted whereas treatment with eicosapentaenoic acid (EPA, 20:5(n-3)) attenuated assembly and secretion of VLDL. Under conditions where the cells were cultured in the presence of 20% serum, EPA (0.4 mM) had marginal effect on the secretion of total apoB100 (determined by pulse-chase analysis) but decreased (by 50%) secretion of triacylglycerol (TG), indicating that the inhibitory effect of EPA was exerted primarily on TG-rich VLDL.

Complete assignments of 1H and 13C NMR spectral data for a novel diterpenoid from Semiaquilegia adoxoides.

Semiaquilegoside A (1), a new diterpenoid, was isolated from the roots of Semiaquilegia adoxoides. Its chemical structure was established as 3beta, 11- dihydroxy-12-O-beta-D-glucopyranosyl-8,11,13-abietatrien-6-one through spectroscopic techniques and chemical analysis.

Influence of perfusion on metabolism and matrix production by bovine articular chondrocytes in hydrogel scaffolds.

Articular cartilage has a limited capacity for self-repair after damage. Engineered cartilage is a promising treatment to replace or repair damaged tissue. The growth of engineered cartilage is sensitive to the extracellular culture environment.

A SU-8/PDMS hybrid microfluidic device with integrated optical fibers for online monitoring of lactate.

A microfluidic device with integrated optical fibres was developed for online monitoring of lactate. The device consists of a SU-8 waveguide, microfluidic channels and grooves for the insertion of optic fibres. It was fabricated by one-step photolithography of SU-8 polymer resist.

Loss of PECAM-1 function impairs alveolarization.

The final stage of lung development in humans and rodents occurs principally after birth and involves the partitioning of the large primary saccules into smaller air spaces by the inward protrusion of septae derived from the walls of the saccules. Several observations in animal models implicate angiogenesis as critical to this process of alveolarization, but all anti-angiogenic treatments examined to date have resulted in endothelial cell (EC) death. We therefore targeted the function of platelet endothelial cell adhesion molecule, (PECAM-1), an EC surface molecule that promotes EC migration and has been implicated in in vivo angiogenesis.