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PubMed2006

Measurement of dimensions of pentagonal doughnut-shaped C-reactive protein using an atomic force microscope and a dual polarisation interferometric biosensor.

In order to develop the C-reactive protein (CRP) sensor chips for clinical detection of atherosclerosis and coronary heart disease, we used an atomic force microscope (AFM) and a dual polarization interferometric (DPI) biosensor to probe the surface ultrastructure and to measure the dimensions of CRP. A single pentagonal structure was directly visualized by AFM, and quantitative measurements of the dimensions of the protein were provided. The average height calculated for each pentagonal CRP particle was approximately 3.03+/-0.37 nm, which basically corresponds to that (36 A in protomer diameter) previously obtained from the structure of CRP determined by X-ray crystallography. Moreover, a experiment using dual polarization interferometric (DPI) as a biosensor was then performed, and the average monolayer thickness value (3.18+/-0.43 nm) that was calculated basically corresponds to that obtained from the experimental value (3.03+/-0.37 nm) of the height measured by an AFM method for CRP. Further investigations will be performed to study the surface ultrastructure of a single pentagonal CRP molecule, and for this purpose a CRP sample (at low concentration) was scanned in vacuum by AFM. The higher-resolution images clearly revealed the presence of doughnut-shaped CRP molecules. In addition, phase images of CRP molecules were captured simultaneously with their height images, and the lateral dimensions of the doughnut-shaped CRP molecules were then measured. It was found that the average values calculated for the outer diameter (11.13+/-1.47 nm) and pore diameter (3.52+/-0.42 nm) are respectively close to those (102 A in outer diameter and 30 A in pore diameter) previously obtained from the structure of CRP determined by X-ray crystallography. This study represents the first direct characterization of the surface ultrastructure and dimensional measurement of the CRP molecule on the sensor chip.

Biosensing Techniques↗
PubMed2006

Interferometric biosensor based on planar optical waveguide sensor chips for label-free detection of surface bound bioreactions.

A label free optical biosensor based on a free-space Young interferometer configuration is presented. Commercial planar Ta(2) O(5) waveguides are used as sensing elements and allow the investigation of surface bound bioreactions like immunoreactions or biological affinity systems. Design criteria are discussed and a detailed characterization of the sensor performance is presented. The developed interferometer yields an effective refractive index resolution of 9 x 10(-9), corresponding to a surface coverage of approximately 13 fg/mm(2). The performance of the system is characterized by two different affinity systems: the antibody-antigen complex protein G-immunoglobulin G is used as a model system for monitoring reaction kinetics. Further measurements on a silanized surface show the formation of a streptavidin monolayer on a biotinylated surface.

Biosensing Techniques↗
PubMed2004

Investigation of initial pellicle formation on modified titanium dioxide (TiO2) surfaces by reflectometric interference spectroscopy (RIfS) in a model system.

OBJECTIVES: Plaque accumulation, leading to inflammatory processes and bone loss, is one of the main reasons for failure of dental implants. Pellicle formation plays a key role in bacterial adhesion and plaque accumulation. The influence of experimental bioactive implant surface coatings on the initial process of pellicle formation was investigated in a model system. METHODS: TiO2-films were modified by covalent binding of laminin and human epidermal growth factor (EGF) to promote adhesion of epithelial cells. Adsorption and dissociation behavior of bovine serum albumin (BSA) and salivary proteins on these surfaces were monitored by time-resolved reflectometric interference spectroscopy (RIfS). RESULTS: The thickness of the irreversibly adsorbed salivary protein layer was reduced from 2.78 +/- 0.71 nm on unmodified TiO2 to 0.78 +/- 0.22 nm on laminin-coated surfaces and to 1.18 +/- 0.29 nm on EGF-coated surfaces. The percentage of initially adsorbed proteins remaining irreversibly bound was reduced from 51 +/- 8% on titanium to 23 +/- 5% by laminin coating and to 44 +/- 11% on EGF-coated surfaces. The highest reduction of protein adsorption (layer thickness lower than 0.05 nm) was achieved on DC-PEG-layers used as spacer for protein coupling. SIGNIFICANCE: Laminin and EGF were shown to be promising candidates for use as biological coatings on the transmucosal part of titanium dental implants where the objective is to enhance epithelial adhesion and inhibit adsorption of salivary proteins and bacteria.

Animals↗
PubMed2005

Mutation of the POU-domain gene Brn4/Pou3f4 affects middle-ear sound conduction in the mouse.

Mutagenesis of the POU-domain gene Brn4/Pou3f4 causes defects in the cochlear duct, semicircular canal, temporal bone and stapes footplate. The footplate defect suggested a middle-ear conductive component to the hearing loss associated with this mutation. This was examined by measuring velocity transfer functions at the umbo of wild type and knockout mice during sound stimulation of the tympanic membrane. When the median umbo velocity of test frequencies in the two groups were compared, the mid-range frequencies of the knockout mice showed a statistically reliable reduction in velocity (maximum of 13 dB) and high variability among animals. These results indicated that mutation of the POU-domain gene, Brn4, changed middle-ear sound conduction when measured at the umbo. The origin of the abnormal velocity response was sought by puncturing a hole in the pars flaccida (PF), and subsequently, measuring movements at the umbo and the head of the long arm of the incus. This hole permitted us to measure velocity at the tip of the incus long arm, just above the incudostapedial joint. The comparison of umbo behavior in both groups with PF perforated showed a loss of sensitivity in the mid-range frequencies of the knockout animals. A comparison of incus velocity in the two groups also exhibited a velocity reduction in the mid-range frequencies of the knockout animals. The reduction at the incus, however, was milder than observed at the umbo. The effect of the perforation in, and variability of, the knockout incus responses may have masked a more potent mid-range frequency effect. Nevertheless, evaluation of the stapes and oval window in knockout mice showed variable pathology from ear to ear. The presence of this pathology, the mid-frequency loss in incus sensitivity and the variability in incus velocity among animals suggested that abnormal stapes behavior in Brn4 deficient mice may determine the response of the ossicles, and thus account for the abnormal mid-frequency umbo behavior seen in knockout animals.

Animals↗
PubMed2004

Evolution of concentration field in a membrane system.

This paper deals with the evolution of concentration field at a single membrane system. Concentration field evolution is described by concentration effect of stable boundary layers, which originate in this system. The concentration effect of boundary layers (CBLE) is studied experimentally on the basis concentration profiles obtained from computer analysis of interferometric pictures of near-membrane regions. Besides experimental results, we also report theoretical investigations and numerical calculations of this effect for two models of membranes (an infinite thin wall and the wall of thickness l). Evolution of concentration field at different distances from membrane surface describes accurately the spatio-temporal structure of the concentration boundary layers (CBLs). Results have shown that their spatial structure is fully established and these layers develop diffusively.

Biological Transport↗
PubMed2026

Structure-informed theoretical modeling defines principles governing avidity in bivalent protein interactions.

In signaling cascades, signaling proteins often encode multiple domains or motifs, which presents the possibility for avidity -- where multivalent binding drastically increases interaction strength and duration. However, predicting and validating multivalent interactions that interact with avidity is a challenge. Here, we integrate mechanistic modeling, structure-based analysis, and experimental approaches as a framework for defining the conditions under which avidity plays a role. We explore the tandem SH2 domain family of interactions with bisphosphorylated partners as a multivalent archetype, which encompasses key secondary messengers in tyrosine kinase signaling networks. Theoretical modeling suggests that maximum avidity occurs with closely spaced tyrosine phosphorylation sites combined with moderate monovalent affinities - exactly around the innate range of SH2 domain affinity - or with phosphorylation sites separated by sufficiently flexible linkers. Surprisingly, despite sequence diversity, structure-based analysis showed relatively conserved three-dimensional spacing between SH2 domains across all tandem SH2 families, which we corroborate experimentally, suggesting evolutionary optimization for avidity interactions. The combination of structure-based analysis of domain spacing with available monovalent experimental data appears, along with iterative experimental refinement of biophysical parameters, can identify high affinity interactions of tandem SH2 domain recruitment to the EGFR C-terminal tail. Using these principles, we extended bivalent predictions into the full phosphoproteome space and structural parameterization of other partners of SH2 domain binding, providing resources and methods for more rapid expansion of bivalent analysis. These approaches lay the groundwork for larger utility in multivalent prediction and testing to help better understand protein interactions that drive cell signaling.

BLI↗
PubMed2004

Axial length and age at cataract surgery.

PURPOSE: To examine the relationship between the axial length of the eye and age at surgery for cataract. SETTING: Postgraduate teaching hospital. METHODS: A retrospective review of a consecutive series of 1776 patients older than 40 years was performed. The patient's age at first-eye surgery, axial length, and preoperative and postoperative best corrected visual acuities were recorded. Linear regression analysis was performed using the age at time of cataract surgery as the dependent variable. RESULTS: Data were available for 1576 of 1670 eyes eligible for inclusion. There was strong evidence of a weak association between a decrease in age at surgery with an increase in axial length (P<.001). The mean patient age at surgery was 74 years for eyes with an axial length less than 25.0 mm, 67 years for eyes with an axial length between 25.0 mm and 30.0 mm, and 61 years for eyes with axial length greater than 30.0 mm. CONCLUSION: An increase in the axial length of the eye was associated with a lower mean age at time of cataract surgery.

Adult↗
PubMed2006

Influence of severity of nuclear cataract on optical biometry.

PURPOSE: To compare preoperative and postoperative measurements of the axial length (AL) performed with the IOLMaster (Carl Zeiss Meditec) to evaluate the effect of nuclear cataract on the optical biometry. SETTING: Department of Ophthalmology, Medical University of Vienna, Vienna, Austria. METHODS: In 245 consecutive eyes of 162 patients scheduled for cataract surgery, type and severity of cataract was evaluated using the Lens Opacities Classification System III. Preoperative and postoperative axial length measurements were performed with the IOLMaster. The postoperative readings of AL were corrected for the IOL optic material implanted. RESULTS: The preoperative AL measurements were 0.07 mm +/- 0.05 (SD) (range -0.18 to 0.11 mm) longer than the postoperative measurements (P<.001). These differences in AL were only weakly correlated (r = -0.28, P = .01) with the nuclear cataract grade, mean 3.5 +/- 0.9. CONCLUSION: In this study, AL measurements with optical biometry using the IOLMaster were not influenced by the severity of nuclear cataract to a clinically relevant degree. Furthermore, it was found that AL in the phakic eye was overestimated by about 0.07 mm because of a slightly too low value of the group refractive index used for the crystalline lens.

Adult↗
PubMed2005

A self-mixing laser-diode interferometer for measuring basilar membrane vibrations without opening the cochlea.

A laser-diode forms the basis of a displacement sensitive homodyne interferometer suitable for measurements from poorly reflective surfaces. The compact and cost-effective interferometer utilizes the self-mixing effect when laser light reflected from a moving target re-enters the laser cavity and causes phase dependent changes of the lasing intensity. A piezo positioner was used to displace the interferometer with known frequency and amplitude as a basis for real-time calibration of the interferometer's sensitivity. The signal-processing algorithm is described that allows measurements in presence of high amplitude noise leading to variation of the interferometer's operating point. Measurements of sound-induced basilar membrane displacements were made in the intact cochleae of rodents by focusing the laser beam of the interferometer through the transparent round window membrane. The interferometer provides a viable means for making subnanometre mechanical measurements from structures in the inner ears of small mammals, where opening of the cochlea is not practicable.

Algorithms↗
PubMed2004

In-plane electronic speckle pattern of interference (ESPI) with optical fibre system applied to the study of the human jaw.

The standard illumination set-up for measurement of in-plane displacement in electronic speckle pattern of interference (ESPI) requires the use of two oblique coherent beams for the illumination of the object. These two beams define a direction in the plane perpendicular to the optical axis in which the displacement can be measured, while in the perpendicular direction the interferometer has no sensitivity. In this paper, a new ESPI system for the measurement of in-plane displacements is used. The novelty presented by this assembly is the use of optical fibres to split and guide the light beams. In this way, it is possible to vary with great precision and ease the direction of the projected beams and, consequently, the direction of the plane in which the displacements undergone by the object can be studied. This set-up makes possible the study of objects with complex geometries, which have deformations with unknown symmetry. We present results obtained with this interferometer applied to the study of the mechanical behaviour of the human jaw.

Algorithms↗
PubMedDate not supplied

Plastic mine detecting radar system using complex-valued self-organizing map that deals with multiple-frequency interferometric images.

Ground penetrating radars (GPR's) have been often applied to underground object imaging. However, conventional radar systems do not work sufficiently to detect anti-personnel plastic landmines. We propose a novel radar imaging system, which processes adaptively interferometric front-end data obtained at multiple-frequency points. The system deals with interferometric images using complex-valued self-organizing map (C-SOM). We demonstrate a successful visualization of a plastic mine buried near the ground surface.

Algorithms↗
PubMed2005

Scanning laser polarimetry retinal nerve fiber layer thickness measurements after LASIK.

PURPOSE: To compare retinal nerve fiber layer (RNFL) thickness measurements before and after LASIK. DESIGN: Cohort study. PARTICIPANTS: Twenty participants undergoing LASIK and 14 normal controls. METHODS: Retinal nerve fiber layer thickness was measured before LASIK and approximately 3 months after surgery in one eye each of 20 patients using a scanning laser polarimeter (GDx Nerve Fiber Analyzer) with fixed corneal compensation (FCC), one with variable corneal compensation (GDx VCC), and optical coherence tomography (OCT). Fourteen normal controls also were tested at baseline and approximately 3 months later. MAIN OUTCOME MEASURES: Retinal nerve fiber layer thicknesses measured with the GDx FCC, GDx VCC, and OCT. RESULTS: At baseline, mean (95% confidence interval [CI]) RNFL thicknesses for the GDx FCC, GDx VCC, and OCT were 78.1 microm (72.2-83.9), 54.3 microm (52.7-56.0), and 96.8 microm (93.2-100.5), respectively. In both LASIK and control groups, there were no significant changes between baseline and follow-up examinations in GDx VCC and OCT RNFL thickness measurements globally or in the superior and inferior quadrants (mean change, <5 microm for each instrument). In the control group, there also was no significant change in GDx FCC measurements between baseline and follow-up. In LASIK patients, significant reductions were observed in GDx FCC RNFL measurements. Average absolute values of the mean (95% CI) change in thickness were 12.4 microm (7.7-17.2), 15.3 microm (9.6-20.9), and 12.9 microm (7.6-18.1) for GDx FCC RNFL measurements superiorly, inferiorly, and globally, respectively (all Ps < or = 0.001). CONCLUSIONS: LASIK does not seem to change RNFL thickness. Reduction in GDx FCC RNFL thickness measurements after LASIK is a measurement artifact and is most likely due to erroneous compensation for corneal birefringence. With scanning laser polarimetry, it is mandatory to compensate individually for change in corneal birefringence after LASIK to ensure accurate RNFL assessment.

Adult↗
PubMed2005

Effects of moderate changes in intraocular pressure on ocular hemodynamics in patients with primary open-angle glaucoma and healthy controls.

PURPOSE: There is some indirect evidence for altered autoregulation in patients with glaucoma, but only a few studies have measured ocular blood flow directly during changes in ocular perfusion pressure. The present study was designed to compare pulsatile choroidal blood flow and optic nerve head (ONH) blood flow during moderate increases in intraocular pressure (IOP) in patients with primary open-angle glaucoma (POAG) and normal controls. DESIGN: Two nonrandomized studies comparing blood flow responses in glaucoma patients and controls in an open design. PARTICIPANTS AND CONTROLS: Sixteen patients with POAG glaucoma and 16 healthy gender-matched and age-matched controls were included in the choroidal blood flow experiments. The ONH blood flow experiment was performed in 14 POAG patients and 14 healthy gender-matched and age-matched controls. METHODS: In the first study, pulsatile choroidal blood flow was assessed by laser interferometric measurement of fundus pulsation amplitude (FPA). In the second study, ONH blood flow was measured using laser Doppler flowmetry. In both cohorts, the IOP was increased stepwise by 10 and 20 mmHg using a suction cup. MAIN OUTCOME MEASURES: Fundus pulsation amplitude and ONH blood flow. RESULTS: The baseline values of FPA and ONH blood flow were lower in glaucoma patients as compared with age-matched and gender-matched healthy controls. In patients with POAG, FPA decreased by -4.5+/-5.8% and -11.3+/-4.9% during elevation of IOP of 10 and 20 mmHg, respectively. These results were not different from the results in healthy controls, where FPA decreased by -5.1+/-3.4% and -12.2+/-4.9% at the 2 pressure levels (P = 0.23 between groups). Optic nerve head blood flow showed no changes during the increase of IOP of 10 and 20 mmHg in either of the 2 groups (glaucoma patients, +2.1+/-14.7% and -0.8+/-15.2%; healthy controls, +4.3+/-12.0% and +0.2+/-14.2%; P = 0.83 between groups). CONCLUSIONS: The present study does not provide evidence for altered autoregulation in patients with POAG during a moderate increase in IOP. However, these results do not necessarily contradict the concept of vascular dysregulation in glaucoma.

Aged↗
PubMed2006

Anterior chamber depth measurement before and after photorefractive keratectomy: comparison between IOL master and Orbscan II.

PURPOSE: To measure the anterior chamber depth (ACD) with 2 different devices before and after photorefractive keratectomy (PRK). DESIGN: Noncomparative case series. PARTICIPANTS: One hundred forty-three eyes of 143 patients who had undergone PRK with refractive errors ranging from -13.13 diopters (D) to +7 D (mean, -3.67+/-3.58) were analyzed. METHODS: The ACD values preoperatively and at 1, 3, and 6 months postoperatively were measured with the Orbscan II and IOL Master. The results were analyzed using the Pearson correlation. MAIN OUTCOME MEASURE: Anterior chamber depth. RESULTS: The instruments showed good agreement between the measurements before and after surgery. A significant decrease between the preoperative and 1-month postoperative measurements was found in the ACD measured from the epithelium with Orbscan II (P<0.01) and IOL Master (P<0.01). A nonsignificant decrease with both IOL Master (P>0.01) and Orbscan II (P>0.01) was found between 3 and 6 months after surgery. The ACD measured from the endothelium using the Orbscan II showed a significant difference only between the 3- and 6-month follow-up data (P<0.01). CONCLUSIONS: The 2 devices showed good agreement, and the changes detected postoperatively seem to be related not only to corneal thinning but also to anterior segment remodeling.

Adolescent↗
PubMed2006

Spectral measurement of the caesium D(2) line with a tunable heterodyne interferometer.

The optical properties of a caesium atomic beam driven on a resonant hyperfine transition in the D(2) line were studied as a function of the probe laser frequency. Using a third off-resonant laser system, a heterodyne interferometer allowed simultaneous absorption and phase shift measurements of either the probe or the coupling laser. The signal features of the probe and coupling laser transmitted intensities showed strong differences in the vicinity of the hyperfine transitions excited by the probe laser. Regular absorption signals and electromagnetically induced transparency were found in either transmitted intensities. Furthermore, light induced birefringence of the probe laser was measured.

Biomechanical Phenomena↗
PubMed2006

Ultrasonic evaluation of residual stresses in flat glass tempering by an original double interferometric detection.

In industrial thermal tempering of glass, the knowledge of the homogeneity of compressive residual stress field on the glass product is fundamental to guarantee the quality of the tempered glass product. In this paper, we use the acoustoelasticity phenomenon in order to estimate the residual stress distribution by using acoustic surface wave. We present an experimental setup based on a double interferometric detection in which an aspheric lens is associated with a beam splitter and a YAG laser whose power is 100 mW. This relative high power enables us to carry out measurements on surface flat glass although optical reflection coefficient is typically weak (< 10%). Using these two points of detection, the evolution of relative surface wave velocity is obtained with a good accuracy. At last, a comparison between the numerical modeling and experimental results shows the potentiality of an ultrasonic method to estimate stress distribution in flat glass tempering.

Algorithms↗
PubMed2005

Effect of myopia on visual acuity measured with laser interference fringes.

The aim was to determine how visual acuity is affected by myopia when optical factors of the eye are controlled. Grating acuity was measured with interference fringes to avoid the effects of aberrations, and ocular biometry was used to compensate for differences in retinal image size among subjects. Distance spectacle refractions ranged from +2.25 to -14.75 D. The retinal magnification factor (RMF) in mm/deg was computed for each eye from the distance refraction, central corneal power and ultrasound biometry. A forced-choice orientation discrimination method was used to measure acuity for high-contrast 543 nm laser interference fringes in three retinal locations: the fovea, and at 4 deg and 10 deg eccentricity in the temporal retina. Acuity, expressed in c/deg and adjusted for spectacle magnification, was not significantly correlated with refraction at any of the three retinal locations. When acuity was converted to retinal spatial frequency units (c/mm) via the RMF, acuity decreased with increasing myopia at all three retinal locations (significantly at the fovea and at 10 deg eccentricity). Retinal acuity values in highly myopic subjects (>6 D) are consistent with retinal sampling distances that are larger than published values of human cone or ganglion cell spacing. The results imply that a highly myopic eye has retinal neurons that are more widely spaced than normal, but the increased axial length enlarges the retinal image enough to compensate for the retinal stretching. The data are consistent with a retinal stretching model that primarily affects the posterior pole.

Adult↗