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PubMed2005

Interactions between sodium dodecyl sulphate and non-ionic cellulose derivatives studied by size exclusion chromatography with online multi-angle light scattering and refractometric detection.

The novel approach described allows to characterise the surfactant-polymer interaction under several sodium dodecyl sulphate (SDS) concentrations (0-20 mM) using size exclusion chromatography (SEC) with online multi-angle light scattering (MALS) and refractometric (RI) detection. Three different cellulose derivatives, hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC) and hydroxyethyl cellulose (HEC), have been studied in solution containing 10 mM NaCl and various concentrations of sodium dodecyl sulphate. It is shown that this approach is well suited for successful application of both Hummel-Dreyer and multi-component light scattering principles and yields reliable molecular masses of both the polymer complex and the polymer itself within the complex, the amount of surfactant bound into the complex as well as appropriate values of the refractive index increment (dn/dc)micro, of both the complex and the polymer in question. The more hydrophobic derivatives HPC and HPMC adsorbed significantly more SDS than HEC. The inter-chain interactions close to critical aggregation concentration (cac) were clearly seen for HPC and HPMC as an almost two-fold average increase in polymer molecular mass contained in the complex.

Cellulose↗
PubMed2005

Microfabricated refractive index gradient based detector for reversed-phase liquid chromatography with mobile phase gradient elution.

Typical refractive index (RI) detectors for liquid chromatography (LC) are not well suited to application with mobile phase gradient elution, due to the difficulty in correcting for the detected baseline shift during the gradient. We report a sensitive, highly reproducible, microfabricated refractive index gradient (micro-RIG) detector that performs well with mobile phase gradient elution LC. Since the micro-RIG signal remains on-scale throughout the mobile phase gradient, one can apply a baseline correction procedure. We demonstrate that by collecting two mobile phase gradient blanks and subtracting one of them from the other, a reproducible, flat baseline is achieved. Therefore, subtracting a blank from a separation provides a baseline corrected chromatogram with reasonably high signal-to-noise ratio for eluting analytes. The micro-RIG detector uses a collimated diode laser beam to optically probe a RIG formed perpendicular to the laminar flow direction within a microfabricated borosilicate glass chip. The chip-based design of the detector is suitable for either traditional bench-top or LC-on-a-chip technologies. We report reversed phase high performance liquid chromatography (RP-HPLC) separations of proteins and polymers, over mobile phase gradient conditions of 67% A:33% B to 3% A:97% B by volume, where A is 96% methanol:3.9% water:0.1% trifluoroacetic acid (TFA), and B is 3.9% methanol:96% water:0.1% TFA. The separations were performed on a Jupiter 5 mu C4 300 A 150 mm x 1.0 mm Phenomenex column at a flow rate of 20 microl/min. Viscosity changes during the mobile phase gradient separation are found to shift the on-chip merge position of the detected concentration gradient (i.e., RIG), in a reproducible fashion. However, this viscosity effect makes detection sensitivity vary throughout the mobile phase gradient, due to moving the optimized position of the probe beam in relation to the analyte concentration gradient being probed. None-the-less, consistent limits of detection (LODs) were achieved. The 3-sigma deflection angle LOD was 16 microrad for micro-RIG detection, corresponding to an injected concentration LOD of 7 ppm (mass/mass) for cytochrome c.

Chromatography, High Pressure Liquid↗
PubMed2006

High-performance liquid chromatographic determination of chitin in the snow crab, Chionoecetes opilio.

A HPLC method was developed for the determination of glucosamine in crab shells. The amount of glucosamine residue can be used to estimate the amount of chitin in analyzed material. Crab samples were hydrolysed with 6M HCl at 100 degrees C for 13 h and the released glucosamine was determined by HPLC using refractive index (RI) detection. Separations were performed using an ODS-2 column with an isocratic mobile phase consisting of 10% methanol, 90% hexanesulfonate, pH 2.1. Glucosamine identification was carried out not only on basis of retention time, but also with respect to mass spectra, which were acquired by HPLC-MS.

Animals↗
PubMed2005

Changes in water content of high plus hydrogel lenses worn on an extended wear basis in a geriatric aphakic population.

PURPOSE: To investigate the water content of hydrogel lenses of relatively high plus power hydrogel lenses after 3 months extended wear and compare values with unused lenses. METHOD: Geriatric aphakic extended wear soft lens patients were fitted with one of four different brands of hydrogel lenses (A) Incanto 78 (Cantor and Nissel, UK), (B) PSL72 (Prospect lenses, UK), (C) ES70 (Ocular Sciences), (D) Proclear (Coopervision). After 3 months continuous wear the lens was removed and water content (WC) was determined at both lens surfaces using an Abbé refractometer. The water content was also measured for 40 unused lenses (+10 to +20D, 10 lenses per brand). RESULTS: One hundred and thirty-two lenses were checked after 3 months extended wear. Mean (+/-S.D.) WC values for front (f) and back (b) surfaces of worn and unworn lenses were, respectively, (A) Worn (n=45): f 73.2(4.13) b 73.8(4.33); unworn f 80.5(0.68) b 81.1 (0.80). (B) Worn (n=37); f 70.5(4.49) b 70.9 (3.89); unworn f 72.5(0.94) b 72.3 (0.89). (C) Worn (n=34); f 68.3(3.18) b 68.4(3.63); unworn f 70.6 (0.48) b 71.1 (0.55). (D) Worn (n=16); f 63.4(3.68) b 63.3(3.19); unworn f 60.9 (1.56) b 61.5 (1.92). There was a significant correlation between WC measured from front and back surfaces (p < 0.01). In worn A-C lenses, front surface WC tended to be lower than back surface WC. For lenses A and C at both surfaces the WC of worn lenses was significantly lower than unworn lenses (p < 0.05). For lens D, mean WC of worn lenses was significantly higher than unworn lenses (p < 0.05). In 80% of B lenses, surface WC of worn lenses were significantly lower than WC of unworn lenses (p < 0.05). There was no relationship between WC and lens power. CONCLUSION: On average, A and C lenses tended to desiccate but D lenses tended to swell as did 20% of B lenses. The front surface of worn lenses measured lower water content than the back surface suggesting the front surface is drier than the back. This apparent difference in water content between the surfaces could be an artefact emanating from differential rates of surface deposition.

Aged↗
PubMed2005

Evaluation of the effect of collagen network degradation on the frictional characteristics of articular cartilage using a simultaneous analysis of the contact condition.

BACKGROUND: Superficial conditions and integrity of collagen network play an important role on the lubrication performance of articular cartilage. In this work, a technique based on the evanescent waves is used for the evaluation of contact condition during friction tests. METHODS: The frictional and superficial characteristics of the normal and degraded articular cartilages with high and low concentration of collagenase were evaluated. The optical apparatus was set in order to decrease the intensity of a light reflected at the interface between a prism and specimens when collagen fibers are found near the interface. FINDINGS: For all conditions, an increase in the attenuation of reflectance as the friction coefficient increases was observed with reasonable correlation. For the specimens degraded with collagenase, low friction and reduced attenuation of reflectance were observed at the beginning of sliding followed by a gradual increase in both friction and attenuation of reflectance. In comparison to the degraded specimens, normal specimens presented high friction at beginning and low friction at the end of test. INTERPRETATION: The superficial conditions and the presence of water at the articular surface play an important role in the lubrication of synovial joints. The ability to retain water for degraded specimens is impaired due to the loss of proteoglycan observed in the histological sections and hence, their low friction observed at the beginning of the test is not sustained for a long time. The use of evanescent waves demonstrated to be very useful in the analysis of the contact condition of articular cartilage.

Animals↗
PubMed2005

Validation the use of refractometer and mathematic equations to measure dietary formula contents for clinical application.

BACKGROUND AND AIMS: Gastric residual volumes are widely used to evaluate gastric emptying for patients receiving enteral feeding, but controversy exists about what constitutes gastric residual volume. We have developed a method by using refractometer and derived mathematical equations to calculate the formula concentration, total residual volume (TRV), and formula volume. In this study, we like to validate these mathematical equations before they can be implemented for clinical patient care. METHODS: Four dietary formulas were evaluated in two consecutive validation experiments. Firstly, dietary formula volume of 50, 100, 200, and 400 ml were diluted with 50 ml water, and then the Brix value (BV) was measured by the refractometer. Secondly, 50 ml of water, then 100 ml of dietary formula were infused into a beaker, and followed by the BV measurement. After this, 50 ml of water was infused and followed by the second BV measurement. The entire procedure of infusing of dietary formula (100 ml) and waster (50 ml) was repeated twice and followed by the BV measurement. RESULTS: The formula contents (formula concentration, TRV, and formula volume) were calculated by mathematical equations. The calculated formula concentrations, TRVs, and formula volumes measured from mathematic equations were strongly close to the true values in the first and second validation experiments (R2>0.98, P<0.001). CONCLUSIONS: Refractometer and the derived mathematical equations may be used to accurately measure the formula concentration, TRV, and formula volume and served as a tool to monitor gastric emptying for patients receiving enteral feeding.

Enteral Nutrition↗
PubMed2005

Optical transillumination tomography with tolerance against refraction mismatch.

Optical transillumination tomography (OT) is a laser-based imaging modality where ballistic photons are used for projection generation. Image reconstruction is therefore similar to X-ray computed tomography. This modality promises fast image acquisition, good resolution and contrast, and inexpensive instrumentation for imaging of weakly scattering objects, such as for example tissue-engineered constructs. In spite of its advantages, OT is not widely used. One reason is its sensitivity towards changes in material refractive index along the light path. Beam refraction artefacts cause areas of overestimated tissue density and blur geometric details. A spatial filter, introduced into the beam path to eliminate scattered photons, will also remove refracted photons from the projections. In the projections, zones affected by refraction can be detected by thresholding. By using algebraic reconstruction techniques (ART) in conjunction with suitable interpolation algorithms, reconstruction artefacts can be partly avoided. Reconstructions from a test image were performed. Standard filtered backprojection (FBP) showed a round mean square (RMS) deviation from the original image of 9.9. RMS deviation with refraction-tolerant ART reconstruction was 0.33 and 0.24, depending on the algorithm, compared to 0.57 (FBP) and 0.06 (ART) in a non-refracting case. In addition, modified ART reconstruction allowed detection of small geometric details that were invisible in standard reconstructions. Refraction-tolerant ART may be the key to eliminating one of the major challenges of OT.

Algorithms↗
PubMed2005

Surface modification of surface sol-gel derived titanium oxide films by self-assembled monolayers (SAMs) and non-specific protein adsorption studies.

Biological events occurring at the implant-host interface, including protein adsorption are mainly influenced by surface properties of the implant. Titanium alloys, one of the most widely used implants, has shown good biocompatibility primarily through its surface oxide. In this study, a surface sol-gel process based on the surface reaction of metal alkoxides with a hydroxylated surface was used to prepare ultrathin titanium oxide (TiOx) coatings on silicon wafers. The oxide deposited on the surface was then modified by self-assembled monolayers (SAMs) of silanes with different functional groups. Interesting surface morphology trends and protein adhesion properties of the modified titanium oxide surfaces were observed as studied by non-specific protein binding of serum albumin. The surface properties were investigated systematically using water contact angle, ellipsometry, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM) measurements. Results showed that the surface sol-gel process predominantly formed homogeneous, but rough and porous titanium oxide layers. The protein adsorption was dependent primarily on the silane chemistry, packing of the alkyl chains (extent of van der Waals interaction), morphology (porosity and roughness), and wettability of the sol-gel oxide. Comparison was made with a thermally evaporated TiOx-Ti/Si-wafer substrate (control). This method further extends the functionalization of surface sol-gel derived TiOx layers for possible titanium alloy bioimplant surface modification.

Adsorption↗
PubMed2005

Properties of galactocerebroside layers transferred to glassy carbon electrodes: effect of an applied electric field.

Galactocerebroside films deposited onto glassy carbon electrodes have been previously studied through the electrochemical response of a redox couple present in solution. Those experiments indicated that the film is inhomogeneous and that there are lipid-free places. In this work, we present experimental results indicating that those bare regions are formed when the electrode is introduced in an aqueous solution, and that the size and/or amount of uncovered domains increase when negative potentials are applied to the film. The experimental techniques employed for these findings are epifluorescence microscopy and ellipsometry.

Carbon↗
PubMed2004

Archer fish learn to compensate for complex optical distortions to determine the absolute size of their aerial prey.

Many animals, including humans, can visually judge the absolute size of objects regardless of changes in viewing distance and thus despite the resulting dramatic differences in the size of the actual retinal images. For animals that have to judge the size of aerial objects from underwater views, this can be a formidable problem; our calculations show that considerable and strongly viewpoint-dependent corrections are needed to compensate for the effects of light refraction. Archer fish face these optical difficulties because they have to shoot down aerial insects over a wide range of horizontal and vertical distances. We show here that these fish can learn to acquire size constancy with remarkable precision and are thus fully capable of taking complex viewpoint dependency into account. Moreover, we demonstrate that archer fish solve the problem not by interpolating within a set of stored views and distances but by learning the laws that connect apparent size with the fish's relative position to the target. This enables the fish to readily judge the absolute sizes of objects from completely novel views.

Animals↗
PubMed2005

How filler properties, filler fraction, sample thickness and light source affect light attenuation in particulate filled resin composites.

OBJECTIVE: It was hypothesized that by standardizing variables such as light sources, filler types and filler surface treatment, it should be possible to use Beer-Lambert's law to predict light absorption in visible light-cured dental composites. METHODS: Mixture of 50 wt% bisGMA and 50 wt% TEGDMA to which a photo-initiator (0.35 wt% champhorquinone) and a co-initiator (0.7 wt% of dimethylaminoethylmethacrylate) was prepared. Three different filler types (HBB, SBB and KU) were added to that mixture in eight different volume percentage. Filler particles were either silane surface treated or not. Specimens were made with thicknesses of 1-5 mm. Total number of 1200 specimens were made for this study. Light transmission was obtained for halogen source and laser lights, which made the number of observations 2400. The absorbance values of the different materials were analyzed in Matlab with respect to the differences in filler fraction and sample thickness. RESULTS: The obtained results revealed that of the two light sources, more light was absorbed by the composite when the laser light was used. Among different filler types, the HBB filler absorbed most light and the KU filler the least. There were significant differences (p < 0.05) in light absorption between all three filler types. SIGNIFICANCE: By comparing the modeled surfaces generated by Matlab for different materials it was possible to determine how different variables such as filler type, filler surface treatment and light source affect light attenuation. The characteristic of incident light affected the light absorbance, meaning that not only the composite's composition needs to be considered in light absorption studies of dental composites.

Absorption↗
PubMed2006

Fraunhofer diffraction of short-fiber-reinforced composites aligned by an electric field.

OBJECTIVES: This study aims to determine if aligned, short-fiber composites are capable of producing Fraunhofer diffraction patterns similar to those of human enamel sections and to validate the mechanism of diffraction by comparing the experimental and theoretical fiber spacings. METHODS: Rectangular composite specimens were filled with short E-glass fibers (120 microm length, 15 microm diameter) to contents of 1, 5, 10, and 25% (vol%). The fibers were oriented perpendicular to the surface normal using an alternating electric field of 0.75 kV/mm. A He-Ne laser was used with thin slices of specimen placed behind a pinhole to produce diffraction patterns. The locations of the diffraction maxima were used to determine the theoretical slit spacings which were compared to the experimental slit spacings determined by microscopy. RESULTS: The specimens produced diffraction patterns analogous to the theoretical Fraunhofer diffraction of light through multiple slits. This was verified by comparing the theoretical spacing of the fibers calculated from the diffraction pattern with the experimental spacing of the fibers determined from the optical micrographs. SIGNIFICANCE: This work has verified that orientation of short-fiber composites using an electric field can yield composites with sufficient order to produce Fraunhofer diffraction patterns that are qualitatively similar to the diffraction patterns of human enamel sections.

Composite Resins↗
PubMed2006

Influence of scattering/absorption characteristics on the color of resin composites.

OBJECTIVES: Optical scattering has been studied because of its important effects on the color and translucency of a material. The objective of this study was to determine the influence of the scattering and absorption characteristics including translucency and opalescence on the CIE color coordinates (L(*), a(*), and b(*)) of resin composites. METHODS: Color and spectral distribution of seven resin composites (14 shades, 1mm thick) were measured in the reflectance and the transmittance modes. Optical constants including scattering coefficient (S), absorption coefficient (K) and light reflectivity (RI) were calculated from the spectral reflectance data using Kubelka's equations. The opalescence parameter (OP) was calculated as the difference in yellow-blue (Deltab(*)) and red-green (Deltaa(*)) coordinates between the reflected and transmitted colors. The translucency parameter (TP) was calculated for the translucency evaluation. Simple correlation among each of CIE L(*), a(*) and b(*) values measured using varied protocols with optical parameters (S, K, RI, OP and TP) was determined, and multiple regression analysis was used to determine the significantly influencing variables on the color coordinates (p<0.05). RESULTS: CIE L(*), a(*), and b(*) values were influenced by S, K, RI, TP and OP values. The CIE L(*) value without backing and that in the transmission mode were highly correlated with S. OP and TP values also influenced CIE L(*) value. The CIE b(*) value was correlated with the OP value when different composites with the same shade designation were compared. SIGNIFICANCE: Since the scattering and absorption characteristics influence the color of resin composites, the size and volume fraction of fillers should be controlled for the best color reproduction, considering the refractive indices of filler and resin matrix.

Absorption↗
PubMed2006

New perfluoroalkylated amphiphilic methacrylates bearing sulfinyl group as monomers for biomedical applications: water content and oxygen permeability of their copolymers with DEGMA.

Perfluoroalkylated methacrylates 7a-c bearing sulfinyl group within a straight-chain ester group, i.e. CH(2)=C(CH(3))CO(2)CH(2)CH(2)S(O)-CH(2)CH(2)CF(2)(CF(2)CF(2))(n)CF(3) (n=1-3) were prepared by two alternative synthetic sequences from 2-[(polyfluoroalkyl)sulfanyl]ethanols HOCH(2)CH(2)SCH(2)CH(2)CF(2)(CF(2)CF(2))(n)CF(3) (n=1-3) in overall yields of 88-91%. Copolymers of 7a-c with diethylene glycol methacrylate (DEGMA) prepared in bulk under radical conditions display high transparency, increased water content and good oxygen permeability properties, which are advantageous for their application in ophthalmology and as prosthetic materials.

Biocompatible Materials↗
PubMed2006

Quantitative estimation of film forming polymer-plasticizer interactions by the Lorentz-Lorenz Law.

Molar refraction as well as refractive index has many uses. Beyond confirming the identity and purity of a compound, determination of molecular structure and molecular weight, molar refraction is also used in other estimation schemes, such as in critical properties, surface tension, solubility parameter, molecular polarizability, dipole moment, etc. In the present study molar refraction values of polymer dispersions were determined for the quantitative estimation of film forming polymer-plasticizer interactions. Information can be obtained concerning the extent of interaction between the polymer and the plasticizer from the calculation of molar refraction values of film forming polymer dispersions containing plasticizer.

Chemistry, Pharmaceutical↗
PubMed2004

Evaluation and comparison of commercially available Aloe vera L. products using size exclusion chromatography with refractive index and multi-angle laser light scattering detection.

Raw materials supplied as Aloe vera L. (sometimes referred to as Aloe barbadensis) samples often contain different composition of low and high molecular weight components when analyzed by size exclusion chromatography. One major reason for variable compositions of commercial A. vera L. materials is that they are produced by different manufacturing techniques. Consistent composition of matter based upon a given standard has been difficult to define. In addition, the method of quantifying and characterization of these commercially available materials has not been agreed upon within the industry. The end user, whether a researcher, a manufacturer, a marketing arm of industry or the consumer, should know that they are receiving a consistent product. A blind study of 32 various A. vera L. samples from different manufacturers, and a prepared sample of fresh A. vera L. gel with the commercial, biologic drug Acemannan Immunostimulanttrade mark, were analyzed for content of high molecular weight (polysaccharides) material by size exclusion chromatography with refractive index detection (SEC/RI) and SEC/RI coupled with multi-angle laser light scattering (MALLS) detection. Results from the SEC/RI analysis showed significant variation in the high molecular weight content, and the MALLS analysis also showed significant variation versus SEC/RI. In addition, HPLC analysis of the anthraquinone content showed that all samples contained significantly less than that of the raw, unwashed aloe gel. The variation of results from all analysis is attributed to differing methods in which the samples were processed by the different manufacturers.

Aloe↗
PubMed2004

Analysis of minor components in olive oil.

Virgin olive oil is well known for its high content of phenolic substances that are thought to have health-promoting properties. These substances also contribute to the distinctive taste of the oil. In this study, tyrosol, vanillic acid, luteolin, and apigenin were identified and quantified by liquid chromatography mass spectrometry (LC-MS). In the seven samples analysed, tyrosol, the most abundant, was in the range of 1.4-29 mg/kg, vanillic acid was in the range of 0.67-4.0 mg/kg, luteolin was in the range of 0.22-7.0 mg/kg, and apigenin was in the range of 0.68-1.6 mg/kg. It was also shown that in olive oil, squalene can be analysed by using a refractive index detector. In the samples analysed, squalene occurred in the range of 3.9-9.6 g/l.

Chromatography, Liquid↗
PubMed2004

Real-Time, label-free monitoring of tumor antigen and serum antibody interactions.

Conventional techniques for the detection of biomolecular interactions can be limited by the need for exogenous labels, time- and labor-intensive protocols, as well as by poor sensitivity levels. A refractometer instrument has been reconfigured to detect biomolecular interactions through changes in surface plasmon resonance (SPR). The binding kinetics and affinity values of anti-NY-ESO-1 monoclonal antibody, ES121, to the cancer-testis antigen NY-ESO-1 were determined according to the surface heterogeneity model and resulted in K(D) values of 1.3x10(-9) and 2.1x10(-10) M. The reconfigured instrument was then used to measure the interaction between tumor antigens and serum antibodies against these antigens in preselected cancer patient sera samples. The tumor antigens assayed included NY-ESO-1, SSX2 and p53, all used as recombinant proteins containing polyhistidine tags. These results demonstrated that the instrument is capable of detecting the binding of serum antibodies from cancer patient sera to immobilized tumor antigens, consistent with those observed previously in ELISA-based experiments. These results demonstrate the potential of SPR technology for the rapid diagnosis and monitoring immune responses.

Antibodies, Monoclonal↗