Analysis of long-term visual quality with numerical 3D ray tracing after corneal crosslinking treatment.

A numerical 3D ray tracing model was used to evaluate the long-term visual effects of two regimens of corneal crosslinking (CXL) treatment of 48 patients with the corneal degeneration keratoconus. The 3D ray tracing analyses were based on corneal elevation data measured by Scheimpflug photography. Twenty-two patients were treated with standard CXL applied uniformly across the corneal surface, whereas 26 patients underwent a customized, refined treatment only at local zones on the cornea (photorefractive intrastromal CXL; PiXL).

Helicobacter pylori Adapts to Chronic Infection and Gastric Disease via pH-Responsive BabA-Mediated Adherence.

The BabA adhesin mediates high-affinity binding of Helicobacter pylori to the ABO blood group antigen-glycosylated gastric mucosa. Here we show that BabA is acid responsive-binding is reduced at low pH and restored by acid neutralization. Acid responsiveness differs among strains; often correlates with different intragastric regions and evolves during chronic infection and disease progression; and depends on pH sensor sequences in BabA and on pH reversible formation of high-affinity binding BabA multimers.

Optimization of an intraocular lens for correction of advanced corneal refractive errors.

Based on numerical 3D ray tracing, we propose a new procedure to optimize personalized intra-ocular lenses (IOLs). The 3D ray tracing was based on measured corneal elevation data from patients who suffered from advanced keratoconus. A mathematical shape description of the posterior IOL surface, by means of a tensor product cubic Hermite spline, was implemented.

Three-dimensional ray-tracing model for the study of advanced refractive errors in keratoconus.

We propose a numerical three-dimensional (3D) ray-tracing model for the analysis of advanced corneal refractive errors. The 3D modeling was based on measured corneal elevation data by means of Scheimpflug photography. A mathematical description of the measured corneal surfaces from a keratoconus (KC) patient was used for the 3D ray tracing, based on Snell's law of refraction.

Cell shape identification using digital holographic microscopy.

We present a cost-effective, simple, and fast digital holographic microscopy method based upon Rayleigh-Sommerfeld backpropagation for identification of the geometrical shape of a cell. The method was tested using synthetic hologram images generated by ray-tracing software and from experimental images of semitransparent spherical beads and living red blood cells. Our results show that, by only using the real part of the back-reconstructed amplitude, the proposed method can provide information of the geometrical shape of the object and at the same time accurately determine the axial position of the object under study.

Assessing bacterial adhesion on an individual adhesin and single pili level using optical tweezers.

Optical tweezers (OT) are a technique that, by focused laser light, can both manipulate micrometer sized objects and measure minute forces (in the pN range) in biological systems. The technique is therefore suitable for assessment of bacterial adhesion on an individual adhesin-receptor and single attachment organelle (pili) level. This chapter summarizes the use of OT for assessment of adhesion mechanisms of both non-piliated and piliated bacteria.

Characterization of the biomechanical properties of T4 pili expressed by Streptococcus pneumoniae--a comparison between helix-like and open coil-like pili.

Bacterial adhesion organelles, known as fimbria or pili, are expressed by gram-positive as well as gram-negative bacteria families. These appendages play a key role in the first steps of the invasion and infection processes, and they therefore provide bacteria with pathogenic abilities. To improve the knowledge of pili-mediated bacterial adhesion to host cells and how these pili behave under the presence of an external force, we first characterize, using force measuring optical tweezers, open coil-like T4 pili expressed by gram-positive Streptococcus pneumoniae with respect to their biomechanical properties.

Methods and estimations of uncertainties in single-molecule dynamic force spectroscopy.

In dynamic force spectroscopy, access to the characteristic parameters of single molecular bonds requires nontrivial measurements and data processing as the rupture forces are found not only to be distributed over a wide range, but are also dependent on the loading rate. The choice of measurement procedure and data processing methods has a considerable impact on the accuracy and precision of the final results. We analyze, by means of numerical simulations, methods to minimize and assess the magnitude of the expected errors for different combinations of experimental and evaluation methods.

Dynamic force spectroscopy of the Helicobacter pylori BabA-Lewis b binding.

The binding strength of the Helicobacter pylori adhesin-receptor complex BabA-ABO/Lewis b has been analyzed by means of dynamic force spectroscopy. High-resolution measurements of rupture forces were performed in situ on single bacterial cells, expressing the high-affinity binding BabA adhesin, by the use of force measuring optical tweezers. The resulting force spectra revealed the mechanical properties of a single BabA-Leb bond.

Physical properties of the specific PapG-galabiose binding in E. coli P pili-mediated adhesion.

Detailed analyses of the mechanisms that mediate binding of the uropathogenic Escherichia coli to host cells are essential, as attachment is a prerequisite for the subsequent infection process. We explore, by means of force measuring optical tweezers, the interaction between the galabiose receptor and the adhesin PapG expressed by P pili on single bacterial cells. Two variants of dynamic force spectroscopy were applied based on constant and non-linear loading force.

Pulsed digital holographic interferometry for dynamic measurement of rotating objects with an optical derotator.

A method for measuring dynamic deformations of rotating objects with pulsed digital holography is described. An optical derotator is used to compensate for the rotation. A CCD camera is used to record two holograms with a short time separation (20 mus).

Comparative study of various endoscopes for pulsed digital holographic interferometry.

A comparison of several endoscopes as object image carriers in pulsed digital holography is presented. Three multicore flexible fiber endoscopes of different spatial resolution and one rigid endoscope are investigated. The four endoscopes are integrated in a setup for the recording of digital holograms on a CCD camera.

Highly Sensitive Pulsed Digital Holography for Built-in Defect Analysis with a Laser Excitation.

A highly sensitive method is presented for noninvasive defect analysis on thin structures with a Q-switched double-pulsed ruby laser with frequency doubling (347 nm). In our research we feature an all-optical arrangement, where a focused laser pulse derived from the same ruby laser (694 nm) acts as a built-in synchronous excitation source for digital holographic interferometry. The recordings are made with a CCD camera for capturing two holograms (two states of the specimen) corresponding to the two UV laser pulses with a short time separation (10-50 mus).

Pulsed digital holography for deformation measurements on biological tissues.

Digital holograms are recorded of biological tissues by use of a Q-switched double-pulsed ruby laser. An image-plane digital holography setup is used with a CCD camera for capturing two holograms with a short time separation (20-800 micros). Subtraction of the phase distribution in two digital holograms yield a fringe phase map that shows the change in deformation of the tissue surface between the recordings.

Simultaneous three-dimensional dynamic deformation measurements with pulsed digital holography.

The three deformation components x, y, z of a vibrating object are measured simultaneously by use of digital holography with a double-pulse ruby laser source. The object is illuminated from three different directions, each optically path matched with three reference beams such that three independent digital holograms are formed and added incoherently in one single CCD image. The optical phase difference between the two recordings taken for each hologram is quantitatively evaluated by the Fourier-transform method so that a set of three phase maps is obtained, representing the deformation along three sensitivity vectors.

Tomographic reconstruction of transient acoustic fields recorded by pulsed TV holography.

Pulsed TV holography combined with computerized tomography (CT) are used to evaluate the three-dimensional distribution of transient acoustic fields in air. Experiments are performed with an electrical discharge between two electrodes as the sound source. Holograms from several directions of the acoustic field are recorded directly onto a CCD detector by use of a double-pulsed ruby laser as the light source.

Dynamic restacking of Escherichia coli P-pili.

P-pili of uropathogenic Escherichia coli mediate the attachment to epithelial cells in the human urinary tract and kidney and therefore play an important role in infection. A better understanding of this mechanism could help to prevent bacteria from spreading but also provides interesting insights into molecular mechanics for future nanotech applications. The helical rod design of P-pili provides an efficient design to withstand hydrodynamic shear forces.

Helicobacter pylori adhesion to carbohydrates.

Adherence of bacterial pathogens to host tissues contributes to colonization and virulence and typically involves specific interactions between bacterial proteins called adhesins and cognate oligosaccharide (glycan) or protein motifs in the host that are used as receptors. A given pathogen may have multiple adhesins, each specific for a different set of receptors and, potentially, with different roles in infection and disease. This chapter provides strategies for identifying and analyzing host glycan receptors and the bacterial adhesins that exploit them as receptors, with particular reference to adherence of the gastric pathogen Helicobacter pylori.

Measurements of the binding force between the Helicobacter pylori adhesin BabA and the Lewis b blood group antigen using optical tweezers.

Helicobacter pylori is a world-wide spread bacterium that causes persistent infections and chronic inflammations that can develop into gastritis and peptic ulcer disease. It expresses several adhesin proteins on its surface that bind to specific receptors in the gastric epithelium. The most well-known adhesin is BabA, which has previously been shown to bind specifically to the fucosylated blood group antigen Lewis b (Leb).

The unfolding of the P pili quaternary structure by stretching is reversible, not plastic.

P pili are protein filaments expressed by uropathogenic Escherichia coli that mediate binding to glycolipids on epithelial cell surfaces, which is a prerequisite for bacterial infection. When a bacterium, attached to a cell surface, is exposed to external forces, the pili, which are composed of approximately 10(3) PapA protein subunits arranged in a helical conformation, can elongate by unfolding to a linear conformation. This property is considered important for the ability of a bacterium to withstand shear forces caused by urine flow.

Physical properties of Escherichia coli P pili measured by optical tweezers.

The mechanical behavior of individual P pili of uropathogenic Escherichia coli has been investigated using optical tweezers. P pili, whose main part constitutes the PapA rod, composed of approximately 10(3) PapA subunits in a helical arrangement, are distributed over the bacterial surface and mediate adhesion to host cells. They are particularly important in the pathogenesis of E.

Optical tweezers based force measurement system for quantitating binding interactions: system design and application for the study of bacterial adhesion.

An optical force measurement system for quantitating forces in the pN range between micrometer-sized objects has been developed. The system was based upon optical tweezers in combination with a sensitive position detection system and constructed around an inverted microscope. A trapped particle in the focus of the high numerical aperture microscope-objective behaves like an omnidirectional mechanical spring in response to an external force.

Influence of peroxisome proliferator-activated receptor alpha on ubiquinone biosynthesis.

The control of ubiquinone biosynthesis by peroxisome proliferators was investigated using peroxisome proliferator activated receptor alpha (PPARalpha)-null mice. Administration of 2-(diethylhexyl)phthalate to control mice resulted in elevated ubiquinone levels in the liver, while dolichol, dolichyl-P and cholesterol concentrations remained unchanged. In PPARalpha-null mice, the level of these lipids were similar to control levels and administration of the peroxisome proliferator did not increase the levels of ubiquinone.

Reduced cholesterol accumulation and improved deficient peroxisomal functions in a murine model of Niemann-Pick type C disease upon treatment with peroxisomal proliferators.

Niemann-Pick type C disease is an inherited disorder characterized by lysosomal accumulation of cholesterol and the mutant gene has recently been identified. The predicted gene product is a transmembrane protein showing homology to proteins involved in the regulation of cholesterol homeostasis, such as 3-hydroxy-3-methylglutaryl-coenzyme A and the sterol regulatory element binding protein cleavage-activating protein. Recent investigations have established a peroxisomal deficiency, which raised the question of whether peroxisomal proliferation could affect this cholesterol-processing error.

Alterations in the biosynthesis of cholesterol, dolichol and dolichyl-P in the genetic cholesterol homeostasis disorder, Niemann-Pick type C disease.

The biosynthesis of cholesterol, dolichol and dolichyl-P were investigated in a murine model of Niemann-Pick type C disease using both in vitro and in vivo systems. In vivo incorporation of [3H]mevalonate into squalene, dolichol and dolichyl-P decreased. The amount of dolichyl-P was elevated due to a decrease in the rate of degradation.