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Investigating porcine pancreatic phospholipase A2 action on vesicles and supported planar bilayers using a quartz crystal microbalance with dissipation.

We present an investigation of the activity of porcine pancreatic phospholipase A2 towards phospholipids. The phospholipids are presented in three different ways, namely as tethered vesicles, intact surface-bound vesicles, and supported planar bilayers (SPBs). The process is followed using a quartz crystal microbalance which measures both the frequency shift and the energy dissipation factor. This technique is very sensitive not only to the mass of the material deposited on the crystal, but also to its viscoelasticity. The breakdown of the phospholipid vesicles and bilayers consequently gives rise to very large signal changes. Enzyme binding is separated from vesicle hydrolysis using nonhydrolyzable ether lipids. Intact and tethered vesicles give rise to the same profile, indicating that direct immobilization of the vesicles does not affect hydrolysis significantly. The data fit well to a Voight-based model describing the change in film structure with time. Initial enzyme binding to intact vesicles is accompanied by a significant increase in layer thickness as well as a decrease in viscosity and shear modulus. This effect, which is less pronounced in SPBs, is probably mainly due to the accumulation of hydrolysis products in the vesicle prior to rupture of the vesicles and release of bound water, since it disappears when lysolipid is included in the vesicles prior to hydrolysis.

Animals↗

Enzyme immobilization on poly(ethylene-co-acrylic acid) films studied by quartz crystal microbalance with dissipation monitoring.

In this study, we use the quartz crystal microbalance with dissipation monitoring (QCM-D) to study the immobilization of the enzyme horseradish peroxidase (HRP) on poly(ethylene-co-acrylic acid) (PEAA) films. The surface polarity of spin-coated PEAA films was varied by heat treatments in air or in a 30% NaOH aqueous solution leading to COOH-depleted or COOH-enriched surfaces, respectively. Two reaction schemes, direct adsorption and amine coupling, were employed for HRP immobilization on the two surfaces. The shifts in frequency and dissipation, Deltaf and DeltaD, measured by QCM-D and the ratio DeltaD/Deltaf were used to evaluate the binding amount and the conformation of the adsorbed enzyme. It is found that HRP immobilized via covalent linkages forms rigid and little dissipative films. In contrast, directly adsorbed HRP films exhibit a highly dissipative structure. HRP-catalyzed oxidation of the 4-chloro-1-naphthol in the presence of H(2)O(2) was used to characterize the catalytic activity of the HRP films. The results show that the enzymatic activity of the covalently immobilized HRP tends to be higher.

Acrylic Resins↗

An electrochemical quartz crystal impedance study on anti-human immunoglobulin G immobilization in the polymer grown during dopamine oxidation at an Au electrode.

The polymeric film grown during dopamine oxidation at an Au electrode was studied as a novel matrix for immobilizing anti-human immunoglobulin G (IgG) via the electrochemical quartz crystal impedance analysis (EQCIA) method. The growth of the polymeric films at Au electrodes during dopamine oxidation in neutral phosphate buffer (pH 7.4) and the immobilization of anti-human IgG into the polymeric films during their growth have been traced at real time. Lysozyme control experiments suggested that anti-human IgG was electrostatically incorporated into the polymeric film. Also, the porosity of the polymeric films has been discussed by measuring the "wet" and "dry" frequency shifts. Compared with a polypyrrole film immobilized with anti-human IgG, the proposed matrix possessed a larger amount of specific binding sites for human IgG by subsequent immunoreaction tests. The association constant of the anti-human IgG immunoreaction was obtained with satisfactory results.

Crystallization↗

Quartz crystal microbalance studies on bilirubin adsorption on self-assembled phospholipid bilayers.

Bilirubin adsorption on self-assembled phospholipid bilayers was studied using quartz crystal microbalance, and factors influencing its adsorption such as pH, temperature, and solution ionic strength were discussed in detail. The results show the amount of adsorbed bilirubin on self-assembled phospholipid bilayers is small at higher temperature and large at higher pH and solution ionic strength, and the adsorption kinetic parameter estimated from the in situ frequency measurement is (1.8+/-0.27)x10(6) M(-1) (mean +/- S.D.). With the present method, the desorption of adsorbed bilirubin caused by human serum albumin and the photoinduced decomposition of adsorbed bilirubin under light illumination were also examined. QCM measurement provides a useful method for monitoring the adsorption/desorption process of bilirubin on self-assembled phospholipid bilayers.

Adsorption↗

Formation of protein molecular imprints within Langmuir monolayers: a quartz crystal microbalance study.

Protein imprinting leading to enhanced rebinding of ferritin to ternary lipid monolayers is demonstrated using a quartz crystal microbalance. Monolayers consisting of cationic dioctadecyldimethylammonium bromide, non-ionic methyl stearate, and poly(ethylene glycol) bearing phospholipids were imprinted with ferritin at the air/water interface of a Langmuir-Blodgett trough and transferred hydrated to hydrophobic substrates for study. This immobilization was shown by fluorescence correlation spectroscopy to significantly hinder any further diffusion of lipids, while rebinding studies demonstrated up to a six-fold increase in ferritin adsorption to imprinted versus control monolayers. A diminished rebinding of ferritin to its imprint was observed through pH reduction to below the protein isoelectric point, demonstrating the electrostatic nature of the interaction. Rebinding to films where imprint pockets remained occupied by the template protein was also minimal. Studies with a smaller acidic protein revealed the importance of the steric influence of poly(ethylene glycol) in forming the protein binding pockets, as albumin-imprinted monolayers showed low binding of ferritin, while ferritin-imprinted monolayers readily accommodated albumin. The controllable structure-function relationship and limitations of this system are discussed with respect to the application of protein imprinting in sensor development as well as fundamental studies of proteins at dynamic interfaces.

Adsorption↗

Microtensile bond strength of a dual-cure resin core material to glass and quartz fibre posts.

OBJECTIVES: To evaluate the microtensile bond strength (microTBS) of a dual-cure resin core material to different regions of fibre posts using different surface treatments. MATERIALS AND METHODS: Twenty-five silica zirconium glass fibre posts (Snowpost) and 25 quartz fibre posts (Aestheti-Plus) were used and randomly divided into five groups according to the surface treatments: (1) no surface treatment (Control) (2) dual-cure bonding agent, Clearfil Liner Bond 2V (LB) (3) LB followed by light curing for 20 s (LB and C) (4) silane coupling bonding agent agent, Clearfil Photobond with Porcelain Bond Activator (PB+PBA) (5) PB+PBA followed by light curing for 20 s (PB+PBA and LC). Treated posts were cemented into artificial post cavities using a dual-cure composite core material (Clearfil DC Core) and cured for 60 s from the top of the cavity. After 24 h storage in water, each specimen was serially sliced into 8, 0.6 x 0.6 mm2-thick beams for the microTBS test. The data were divided into three regions (upper/middle/bottom) and analyzed using three-way ANOVA and Dunnet's T3 multiple comparisons (alpha=0.05). RESULTS: There were no significant differences in bond strength between the three regions (p>0.05). The bond strengths were enhanced by the application of a silane coupling agent. For Snowpost, photoirradiation of the dual-cure bonding agent applied to the post surface significantly improved the bond strength (p<0.05) whereas it did not affect the bond strength of Aestheti-Plus post (p>0.05). CONCLUSION: The bond strength between fibre post and dual-cure resin core material depends upon the type of post and surface treatment. There were no regional differences in microTBS of the bonded post.

Composite Resins↗

Limits of thermoluminescence dosimetry using quartz extracted from recent building materials in urban settlements.

The luminescence of quartz extracted from recently fired building material is known to detect doses of few mGy and can be successfully employed in the case of large scale radiation exposures due to nuclear accidents or terrorist acts. One brick and two tiles (50-80 years range) collected from an urban settlement were used to test the realistic minimum detectable dose limits of retrospective luminescence dosimetry. Independent methods like alpha counting, beta dosimetry, gamma spectrometry and flame photometry were used for the annual dose assessment. Two approaches were employed for the evaluation of the total accrued dose: regenerative dose and additive dose. The former allows the assessment of doses due to anthropogenic sources of radiation as low as approximately 12 mGy by using 10-year-old samples, but it can be applied only in samples showing no sensitisation. The latter can be applied to any sample, however, the related uncertainty is higher and minimum detectable anthropogenic dose in young samples amounts to 20 mGy.

Cities↗

Extending the chronology of deposits at Blombos Cave, South Africa, back to 140 ka using optical dating of single and multiple grains of quartz.

Optically stimulated luminescence (OSL) measurements are reported for both single aliquots (of two different sizes) and single grains of quartz from deposits within Blombos Cave. Ages have been obtained for six sediments from the Middle Stone Age (MSA) occupation levels and for two sterile sands, one underlying the archaeological sediment and one overlying the Later Stone Age occupation levels. The ages for the archaeological sediments were obtained from single-grain measurements that enabled unrepresentative grains to be rejected. The MSA occupation levels have ages that, within error limits, are in stratigraphic order and fall between the OSL age for the oldest dune sand (143.2+/-5.5 ka) and a previously published OSL age for the sterile sand ( approximately 70 ka) that separates the Middle and Later Stone Age deposits. The earliest MSA archaeological phase, M3, from where fragments of ochre were found as well as human teeth, is dated to 98.9+/-4.5 ka, coinciding with the sea-level high of oxygen isotope substage 5c. The cave then appears to be unoccupied until oxygen isotope substage 5a on the basis of four OSL ages for archaeological phase M2, ranging from 84.6+/-5.8 to 76.8+/-3.1 ka; these levels contained large hearths and bone tools. An age of 72.7+/-3.1 ka was obtained for the final MSA archaeological phase, M1, from which deliberately engraved ochre and shell beads were recovered along with bifacial stone points. We conclude that the periods of occupation were determined by changes in sea level, with abundant sources of seafood available in times of high sea level and with the cave being closed by the accumulation of large dunes during periods of low sea level, such as during oxygen isotope stages 4 and 6.

Chronology as Topic↗

Electrochemical and piezoelectric quartz crystal detection of antisperm antibody based on protected Au nanoparticles with a mixed monolayer for eliminating nonspecific binding.

The detection of antisperm antibody (AsAb) by electrochemical method based on Au nanoparticles with a mixed monolayer for eliminating nonspecific binding is presented. Impedance spectroscopy is used to characterize the modified procedures, the immobilization of sperm antigen (SpAg) and binding of AsAb in the presence of [Fe (CN)(6)](3-/4-). In addition, piezoelectric quartz crystal, PQC, was used to analyze the immobilization quantity of Au nanoparticles as well as to optimize experimental conditions. The change of electron-transfer resistance was correlated with the concentration of AsAb in a range from 25 to 600 mU/ml with the detection limit of 10 mU/ml. The analytical results of several human serum samples using the developed technique are in satisfactory agreement with those given by the enzyme-linked immunosorbent assay (ELISA) method.

Adsorption↗

Methodological aspects of the induction of biofilms for remediation of the visual impact generated by quartz mining.

The opening up of mine faces causes severe alteration to the environment that must be remedied. In the case of quartz mines, the bright white colour of the mineral gives rise to a significant impact that is visible for long distances. New techniques to correct this visual impact, based on the induction of biofilms on the faces, have recently been developed in an attempt to imitate natural effects that mask the impact. Although techniques have been applied successfully in the laboratory, they must be optimized to ensure their success in the field. In the present study, the results of a randomized factorial experiment indicated that although the biofilms adhere to the faces naturally, without the use of adhesives, continued development of the film depends to a large extent on the input of nutrients. Water must also be provided with nutritional supplement to maintain the required level of moisture.

Biofilms↗

Ultrasonic monitoring of yoghurt formation by using AT-cut quartz: lighting of casein micelles interactions process during the acidification.

The behavior of weak gels during their formation singularly attracts attention of dairy products factories. In our study we investigate acidified pre-heated milk gels formation that are fairly often used to product yoghurts. The gel formation requires a tight control of the first step of micelles modification process and the kinetics reaction parameters. The most current rheological parameters used to achieve the monitoring are the storage G' and the loss G'' shear moduli and the gelation time. The study of these parameters is commonly performed at very low frequencies (1 Hz). Our technique uses a 6 MHz AT-cut quartz crystal immersed in an acidified milk solution kept at a constant temperature. This method is singularly effective to ensure a complete and a reliable follow-up of the viscoelastic parameters of casein gels. A suitable new model enables a complete follow-up of the micelles evolution from the viscoelastic properties. The experimental results of the G' and G'' moduli versus temperature and versus glucono-delta-lactone (GDL) added to milk are analyzed. In order to understand the micelles modifications, an analysis of the viscoelastic evolution try to explain the validity of the various models of micelles modification. In addition a new accurate kinetics characteristic time is proposed. This time corresponds to the moment for which the elastic effect of material becomes significant. From the kinetics study of casein gels at various temperatures, the Arrhenius relationship and a modified Flory-Stockmayer relationship give us access to the activation energy. By using the proposed technique and the suitable models developed, the structure thus quality of dairy products may be better controlled.

Algorithms↗

Biological and chemical interactions with U(VI) during anaerobic enrichment in the presence of iron oxide coated quartz.

Microcosm experiments were performed to understand chemical and biological interactions with hexavalent uranium (U(VI)) in the presence of iron oxide bearing minerals and trichloroethylene (TCE) as a co-contaminant. Interactions of U(VI) and hydrous iron oxide moieties on the mineral oxide surfaces were studied during enrichments for dissimilatory iron reducing (DIRB) and sulfate reducing bacteria (SRB). Microbes enriched from groundwater taken from the Test Area North (TAN) site at the Idaho National Laboratory (INL) were able to reduce the U(VI) in the adsorption medium as well as the iron on quartz surfaces. Early in the experiment disappearance of U(VI) from solution was a function of chemical interactions since no microbial activity was evident. Abiotic removal of U(VI) was enhanced in the presence of carbonate. As the experiment proceeded, further removal of U(VI) from solution was associated with the fermentation of lactate to propionate and acetate. During later phases of the experiment when lactate was depleted from the growth medium in the microcosm containing the DIRB enrichments, U(VI) concentrations in the solution phase increased until additional lactate was added. When additional lactate was added and fermentation proceeded, U(VI) concentrations in the liquid phase again returned to near zero. Similar results were shown for the SRB enrichment but lower uranium concentrations were seen in the liquid phase, while in the enrichment with carbonate a similar increase in uranium concentration was not seen. Chemical and biological interactions appear to be important on the mobilization/immobilization of U(VI) in an iron oxide system when TCE is present as a co-contaminant. Interestingly, TCE present in the microcosm experiments was not dechlorinated which was probably an effect of redox conditions that were unsuitable for reductive dechlorination by the microbial culture tested.

Adsorption↗

Surface specific kinetics of lipid vesicle adsorption measured with a quartz crystal microbalance.

We have measured the kinetics of adsorption of small (12.5-nm radius) unilamellar vesicles onto SiO2, oxidized gold, and a self-assembled monolayer of methyl-terminated thiols, using a quartz crystal microbalance (QCM). Simultaneous measurements of the shift in resonant frequency and the change in energy dissipation as a function of time provide a simple way of characterizing the adsorption process. The measured parameters correspond, respectively, to adsorbed mass and to the mechanical properties of the adsorbed layer as it is formed. The adsorption kinetics are surface specific; different surfaces cause monolayer, bilayer, and intact vesicle adsorption. The formation of a lipid bilayer on SiO2 is a two-phase process in which adsorption of a layer of intact vesicles precedes the formation of the bilayer. This is, to our knowledge, the first direct evidence of intact vesicles as a precursor to bilayer formation on a planar substrate. On an oxidized gold surface, the vesicles adsorb intact. The intact adsorption of such small vesicles has not previously been demonstrated. Based on these results, we discuss the capacity of QCM measurements to provide information about the kinetics of formation and the properties of adsorbed layers.

Adsorption↗

Novel quartz flow-cell as a post-column photochemical reactor for high-performance liquid chromatography.

The construction of a new post-column photochemical reactor with quartz flow cells in series for high-performance liquid chromatography (HPLC) is described. The performance of the new reactor was compared with a conventional open tubular PTFE coil reactor. The sensitivity, accuracy and precision obtained with both reactors are comparable. The new reactor has the obvious advantages of smaller cell volume as well as inertness and resistance to not only light and heat produced by the UV lamp, but also to organic solvents in the mobile phases, which results in greatly improved durability, reduced peak broadening and shorter chromatographic run times. Application of the new reactor to the fluorescence detection of DU-6859a, a new fluoroquinolone antimicrobial agent, in human serum is reported.

Anti-Infective Agents↗

Characterization of a human pancreatic secretory trypsin inhibitor mutant binding to Legionella pneumophila as determined by a quartz crystal microbalance.

We describe the isolation from a large phagemid library of a human pancreatic secretory trypsin inhibitor (hPSTI) mutant that binds to Legionella pneumophila. To gain further insight into the binding kinetics of the isolated hPSTI mutant, an immunosensing system based on a quartz crystal microbalance (QCM) was used. In contrast to ELISA procedures, k(on) and k(off) rates could be derived from the QCM sensograms. Thus, it is possible to characterize specific intermolecular interactions between proteins and phages isolated from large phage display libraries by QCM.

Bacterial Outer Membrane Proteins↗

Sorption of lambda-cyhalothrin, cypermethrin, deltamethrin and fenvalerate to quartz, corundum, kaolinite and montmorillonite.

Sorption to mineral surfaces may be important for retention and degradation of hydrophobic pesticides in subsoils and aquifers poor in organic matter. In this work the title pyrethroids have been used to investigate selective interactions with the surfaces of four minerals. Sorption of the four pyrethroids was quantified in batch experiments with initial pyrethroid concentrations of 1-100 microg/l. Sorption to centrifuge tubes used in the batch experiments accounted for 25-60% of total sorption. Net sorption was obtained from total sorption after subtracting the amounts of pyrethroids sorbed to centrifuge tubes used. All isotherms could be fitted by the Freundlich equation with n ranging between 0.9 and 1.1. Bonding affinities per unit surface area decreased in the order: corundum > quartz > montmorillonite approximately equal kaolinite. A similar sequence as found for the total surface tension of the minerals. All minerals showed the same selectivity order with respect to sorption affinity of the four pyrethroids: lambda-cyhalothrin > deltamethrin > cypermethrin > fenvalerate, which shows that the most hydrophobic compound is sorbed most strongly. Stereochemical properties of the four pyrethroid formulations may also contribute to the selectivity pattern.

Adsorption↗

Characterization of heparin surfaces using photoelectron spectroscopy and quartz crystal microbalance.

In order to gain insight into the molecular structure of a new heparin surface (Corline heparin surface, CHS), quartz crystal microbalance dissipation (QCM-D) and photoelectron spectroscopy (PES) were used. Two surfaces, with different surface thickness and surface concentration of heparin, were compared. QCM-D measurements provided information on the thickness of the two different heparin surfaces and confirmed that the process of formation of the two layers were virtually identical. PES showed that the thicker coating resulted in complete coverage of the substrate, whereas with the thinner coating Fe signals originating from the stainless steel substrate could be detected. Information of the molecular surface structure of the heparin coatings could be derived from the ratios at different photon energies between S2p signals from sulfonate groups in heparin and disulfide groups originating from a cross-linking unit in the heparin conjugate.

Biophysics↗