Separation of neutral compounds by high speed capillary electrochromatography

With using short capillary column packed with porous and non-porous ODS stationary phases, high speed separation of 6 neutral aromatic compounds within 36 s by capillary electrochromatography (CEC) has been performed. Good reproducibility of the migration times for those solutes in high speed CEC was observed with RSD less than 1%. Both the linear velocity of EOF and the current linearly increases with the applied voltage, which means that the thermal effect by Joule heating was small. However, the capacity factor of solutes was found to decrease with the increase of the applied voltage, which was caused by the fact that about several seconds needed for the increase of voltage from 0 to applied value on a commercial CE instrument made larger contributions to the migration times of the early eluted compounds than those of lately eluted ones during high speed CEC, and voltage effect would increase with the higher applied voltage used. The linear relationship between the logarithm of capacity factor and the number of carbon for homologous compounds was observed, and positive value of slope means that the hydrophobicity of solutes is one of the main contribution factors to retention in high speed CEC packed with ODS stationary phases.

Synthesis of NaA zeolite membrane by microwave heating

The synthesis of NaA zeolite membrane on a porous alpha -Al2O3 support by microwave heating (MH) was investigated. The formation of a NaA zeolite membrane was drastically promoted by MH. The synthesis time was reduced from 3 h for conventional heating (CH) to 15 min for MH. Surface seeding cannot only promote the formation of NaA zeolite on the support, but also inhibit the transformation of NaA zeolite into other types of zeolites. The thickness of the NaA zeolite membrane synthesized by MH was about 4 mum, thinner than that of NaA zeolite membrane synthesized by CH. The permeance of NaA zeolite membrane synthesized by MH was four times higher than that of the NaA zeolite membrane synthesized by CH, while their permselectivities were comparable. Multi-stage synthesis resulted in the transformation of NaA zeolite into other types of zeolites, and the perfection of the as-synthesized membrane decreased.

Synthesis and characterization of gallium-based perovskite-type dense membrane with oxygen semipermeability

La0.15Sr0.85Ga0.3Fe0.7O3-delta (LSGFO) and La0.15Sr0.85Co0.3Fe0.7O3-delta (LSCFO) mixed oxygen-ion and electron conducting oxides were synthesized by using a combined EDTA and citrate complexing method, and the corresponding dense membranes were fabricated. The properties of the oxide powders and membranes were characterized with combined SEM, XRD, H-2-TPR, O-2-TPD techniques, mechanical strength and oxygen permeation measurement. The results showed that LSGFO had much higher thermochemical stability than LSCFO due to the higher valence stability of Ga3+. After the temperature-programmed reduction by 5% H-2 in Ar from 20 degreesC to 1020 degreesC, the basic perovskite structure of LSGFO was successfully preserved. LSGFO also favors the oxygen vacancy formation better than LSCFO. Oxygen permeation measurement demonstrated that LSGFO had higher oxygen permeation flux than LSCFO, but they had similar activation energy for oxygen transportation, with a value of 110 and 117 kJ . mol(-1), respectively The difference in oxygen permeation fluxes was correlated with the difference in oxygen vacancy concentrations for the two materials.

A novel zirconium-based perovskite-type membrane material for oxygen permeation

A novel zirconium-based membrane material of BaCo0.4Fe0.4Zr0.2O3-6 with cubic perovskite structure was synthesized for the first time through a method of citric and EDTA acid combined complexes. The structural stability was characterized by XRD, O-2-TPD and H-2-TPR techniques respectively. The high oxygen permeation flux of 0.873 mL/cm(2) min at 950 degreesC was obtained under He/Air gradient. Meanwhile, the single activation energy for oxygen permeation and the long-term steady operation of 200 h at 800 degreesC were achieved.

Preparation and characterization of Pd-Ag alloy composite membrane with magnetron sputtering

A Pd-Ag (24 wt%) alloy composite membrane was prepared by the magnetron sputtering. A gamma-Al2O3 membrane was synthesized by the sol-gel method and used as substrate of the Pd-Ag alloy film. The process parameters of the magnetron sputtering were optimized as a function of the compactness of the Pd-Ag alloy film. The best membrane with a thickness of 1 mu m was produced with a sputtering pressure of 2.7 Pa and a substrate temperature of 400 degrees C. The membrane had an H-2/N-2 permselectivity of 51.5-1000 and an H-2 permeation rate of 0.036-1.17 x 10(-5)cm(3)/cm(2).s. Pa, depending on operating conditions.

Synthesis of zeolite NaA membranes with high permeance under microwave radiation on mesoporous-layer-modified macroporous substrates for gas separation

This article reported the NaA zeolite membranes with high permeance synthesized with microwave heating method under different conditions: (1) on a macroporous substrate in gel, (11) on a mesoporous/macroporous (top-mesoporous-layer-modified macroporous) substrate in gel, and (111) on a mesoporous/macroporous substrate in sol. In general, the H-2 permeance of the NaA membranes by microwave heating in gel was usually at the level of 10(-6) mol s(-1) m(-2) Pa-1, much higher than that by the conventional hydrothermal synthesis. At similar H-2/C3H8 permselectivity. On the substrate modified mesoporous top layer, the H-2 permeance of the NaA membranes by microwave heating in gel or sol was further enhanced, while maintaining comparable H-2/C3H8 permselectivity, due to the prevention of penetration of the reagent into the pores of the macroporous substrate. Meanwhile, the synthesis took less time in sol than in gel on the mesoporous/macroporous substrate. The NaA membranes synthesized in sol had larger permeance than those in gel and underwent transformation in shorter time. The permeation of C3H8 suggested that there existed unwanted intercrystalline pores or defects in the membranes. © 2005 Elsevier B.V. All rights reserved.

Development of a LCST membrane forming system for cellulose acetate ultrafiltration hollow fiber

A novel lower critical solution temperature (LCST) membrane forming system containing cellulose acetate (CA)/poly (vinyl pyrrolidone) (PVP 3 60K)/N-methyl-2-pyrrolidone (NMP)/1,2-propanediol with a weight ratio of 24.0:5.0:62.6:8.4 had been developed. CA hollow fiber ultrafiltration (UF) membranes were fabricated using the dry-wet spinning technique. The fibers were post-treated with a 200 mg/L hypochlorite solution over a period of 6 It at pH 7. The experimental results showed that water flux of a membrane decreased while retention increased with increasing CA concentration in a dope. It was concluded that the membrane pore size decreased with increasing CA concentration. The membrane fouling tendency for BSA was 3 times higher than that for PVP 24K. (C) 2004 Elsevier B.V. All rights reserved.

Advanced polymer membrane development in pervaporation dehydration and lateral flow diagnostics

The work in this thesis concerns the advanced development of polymeric membranes of two types; pervaporation and lateral-flow. The former produced from a solution casting method and the latter from a phase separation. All membranes were produced from casting lacquers. Early research centred on the development of viable membranes. This led to a supported polymer blend pervaporation membrane. Selective layer: plasticized 4:1 mass ratio sodium-alginate: poly(vinyl-alcohol) polymer blend. Using this membrane, pervaporation separation of ethanol/water mixtures was carefully monitored as a function of film thickness and time. Contrary to literature expectations, these films showed increased selectivity and decreased flux as film thickness was reduced. It is argued that morphology and structure of the polymer blend changes with thickness and that these changes define membrane efficiency. Mixed matrix membrane development was done using spherical, discreet, size-monodisperse mesoporous silica particles of 1.8 - 2μm diameter, with pore diameters of ~1.8 nm were incorporated into a poly(vinyl alcohol) [PVA] matrix. Inclusion of silica benefitted pervaporation performance for the dehydration of ethanol, improving flux and selectivity throughout in all but the highest silica content samples. Early lateral-flow membrane research produced a membrane from a basic lacquer composition required for phase inversion; polymer, solvent and non-solvent. Results showed that bringing lacquers to cloud point benefits both the pore structure and skin layers of the membranes. Advancement of this work showed that incorporation of ethanol as a mesosolvent into the lacquer effectively enhances membrane pore structure resulting in an improvement in lateral flow rates of the final membranes. This project details the formation mechanics of pervaporation and lateral-flow membranes and how these can be controlled. The principle methods of control can be applied to the formation of any other flat sheet polymer membranes, opening many avenues of future membrane research and industrial application.

HER2: a 'predictive factor' ready to use in the daily management of breast cancer patients?

The past few years have witnessed an exponential increase in studies trying to identify molecular markers in patients with breast tumours that might predict for the success or failure of hormonal therapy or chemotherapy. HER2, a tyrosine kinase membrane receptor of the epidermal growth factor receptor family, has been the most widely studied marker in this respect. This paper attempts to critically review to what extent HER2 may improve 'treatment individualisation' for the breast cancer patient. Copyright (C) 2000.

Growth factor erv1-like modulates Drp1 to preserve mitochondrial dynamics and function in mouse embryonic stem cells.

The relationship of mitochondrial dynamics and function to pluripotency are rather poorly understood aspects of stem cell biology. Here we show that growth factor erv1-like (Gfer) is involved in preserving mouse embryonic stem cell (ESC) mitochondrial morphology and function. Knockdown (KD) of Gfer in ESCs leads to decreased pluripotency marker expression, embryoid body (EB) formation, cell survival, and loss of mitochondrial function. Mitochondria in Gfer-KD ESCs undergo excessive fragmentation and mitophagy, whereas those in ESCs overexpressing Gfer appear elongated. Levels of the mitochondrial fission GTPase dynamin-related protein 1 (Drp1) are highly elevated in Gfer-KD ESCs and decreased in Gfer-overexpressing cells. Treatment with a specific inhibitor of Drp1 rescues mitochondrial function and apoptosis, whereas expression of Drp1-dominant negative resulted in the restoration of pluripotency marker expression in Gfer-KD ESCs. Altogether, our data reveal a novel prosurvival role for Gfer in maintaining mitochondrial fission-fusion dynamics in pluripotent ESCs.

beta2-Adrenergic receptor regulation by GIT1, a G protein-coupled receptor kinase-associated ADP ribosylation factor GTPase-activating protein.

G protein-coupled receptor activation leads to the membrane recruitment and activation of G protein-coupled receptor kinases, which phosphorylate receptors and lead to their inactivation. We have identified a novel G protein-coupled receptor kinase-interacting protein, GIT1, that is a GTPase-activating protein (GAP) for the ADP ribosylation factor (ARF) family of small GTP-binding proteins. Overexpression of GIT1 leads to reduced beta2-adrenergic receptor signaling and increased receptor phosphorylation, which result from reduced receptor internalization and resensitization. These cellular effects of GIT1 require its intact ARF GAP activity and do not reflect regulation of GRK kinase activity. These results suggest an essential role for ARF proteins in regulating beta2-adrenergic receptor endocytosis. Moreover, they provide a mechanism for integration of receptor activation and endocytosis through regulation of ARF protein activation by GRK-mediated recruitment of the GIT1 ARF GAP to the plasma membrane.

Outer Membrane Vesicle Production in Escherichia coli Relieves Envelope Stress and is Modulated by Changes in Peptidoglycan

Bacterial outer membrane vesicles (OMVs) are spherical buds of the outer membrane (OM) containing periplasmic lumenal components. OMVs have been demonstrated to play a critical part in the transmission of virulence factors, immunologically active compounds, and bacterial survival, however vesiculation also appears to be a ubiquitous physiological process for Gram-negative bacteria. Despite their characterized biological roles, especially for pathogens, very little is known about their importance for the originating organism as well as regulation and mechanism of production. Only when we have established their biogenesis can we fully uncover their roles in pathogenesis and bacterial physiology. The overall goal of this research was to characterize bacterial mutants which display altered vesiculation phenotypes using genetic and biochemical techniques, and thereby begin to elucidate the mechanism of vesicle production and regulation. One part of this work elucidated a synthetic genetic growth defect for a strain with reduced OMV production (ΔnlpA, inner membrane lipoprotein with a minor role in methionine transport) and envelope stress (ΔdegP, dual function periplasmic chaperone/ protease responsible for managing proteinaceous waste). This research showed that the growth defect of ΔnlpAΔdegP correlated with reduced OMV production with respect to the hyprevesiculator ΔdegP and the accumulation of protein in the periplasm and DegP substrates in the lumen of OMVs. We further demonstrated that OMVs do not solely act as a stress response pathway to rid the periplasm of otherwise damaging misfolded protein but also of accumulated peptidoglycan (PG) fragments and lipopolysaccharide (LPS), elucidating OMVs as a general stress response pathway critical for bacterial well-being. The second part of this work, focused on the role of PG structure, turnover and covalent crosslinks to the OM in vesiculation. We established a direct link between PG degradation and vesiculation: Mutations in the OM lipoprotein nlpI had been previously established as a very strong hypervesiculation phenotype. In the literature NlpI had been associated with another OM lipoprotein, Spr that was recently identified as a PG hydrolase. The data presented here suggest that NlpI acts as a negative regulator of Spr and that the ΔnlpI hypervesiculation phenotype is a result of rampantly degraded PG by Spr. Additionally, we found that changes in PG structure and turnover correlate with altered vesiculation levels, as well as non-canonical D-amino acids, which are secreted by numerous bacteria on the onset of stationary phase, being a natural factor to increase OMV production. Furthermore, we discovered an inverse relationship between the concentration of Lpp-mediated, covalent crosslinks and the level of OMV production under conditions of modulated PG metabolism and structure. In contrast, situations that lead to periplasmic accumulation (protein, PG fragments, and LPS) and consequent hypervesiculation the overall OM-PG crosslink concentration appears to be unchanged. Form this work, we conclude that multiple pathways lead to OMV production: Lpp concentration-dependent and bulk driven, Lpp concentration-independent.

Modulation of bacterial outer membrane vesicle production by envelope structure and content.

BACKGROUND: Vesiculation is a ubiquitous secretion process of Gram-negative bacteria, where outer membrane vesicles (OMVs) are small spherical particles on the order of 50 to 250 nm composed of outer membrane (OM) and lumenal periplasmic content. Vesicle functions have been elucidated in some detail, showing their importance in virulence factor secretion, bacterial survival, and biofilm formation in pathogenesis. Furthermore, OMVs serve as an envelope stress response, protecting the secreting bacteria from internal protein misfolding stress, as well as external envelope stressors. Despite their important functional roles very little is known about the regulation and mechanism of vesicle production. Based on the envelope architecture and prior characterization of the hypervesiculation phenotypes for mutants lacking the lipoprotein, Lpp, which is involved in the covalent OM-peptidoglycan (PG) crosslinks, it is expected that an inverse relationship exists between OMV production and PG-crosslinked Lpp. RESULTS: In this study, we found that subtle modifications of PG remodeling and crosslinking modulate OMV production, inversely correlating with bound Lpp levels. However, this inverse relationship was not found in strains in which OMV production is driven by an increase in "periplasmic pressure" resulting from the accumulation of protein, PG fragments, or lipopolysaccharide. In addition, the characterization of an nlpA deletion in backgrounds lacking either Lpp- or OmpA-mediated envelope crosslinks demonstrated a novel role for NlpA in envelope architecture. CONCLUSIONS: From this work, we conclude that OMV production can be driven by distinct Lpp concentration-dependent and Lpp concentration-independent pathways.