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At least 19 records

Functional role for stable microtubules in lens fiber cell elongation

Highlights: • N-cadherin junctions link to stable microtubules in elongating lens fiber cells. • Stabilized microtubules are essential to the process of fiber cell elongation. • Formation/stabilization of epithelial fiber interface is microtubule dependent. • Loss of stable microtubules in the developing lens induces myosin activation. • Disassembly of stable microtubules alters N-cadherin junction assembly. • Stable microtubules required for normal lens fiber cell morphogenesis. The process of tissue morphogenesis, especially for tissues reliant on the establishment of a specific cytoarchitecture for their functionality, depends a balanced interplay between cytoskeletal elements and their interactions with cell adhesion molecules. The microtubule cytoskeleton, which has many roles in the cell, is a determinant of directional cell migration, a process that underlies many aspects of development. We investigated the role of microtubules in development of the lens, a tissue where cell elongation underlies morphogenesis. Our studies with the microtubule depolymerizing agent nocodazole revealed an essential function for the acetylated population of stable microtubules in the elongation of lens fiber cells, which was linked to their regulation of the activation state of myosin. Suppressing myosin activation with the inhibitor blebbistatin could attenuate the loss of acetylated microtubules by nocodazole and rescue the effect of this microtubule depolymerization agent on both fiber cell elongation and lens integrity. Our results also suggest that acetylated microtubules impact lens morphogenesis through their interaction with N-cadherin junctions, with which they specifically associate in the region where lens fiber cell elongate. Disruption of the stable microtubule network increased N-cadherin junctional organization along lateral borders of differentiating lens fiber cells, which was prevented by suppression of myosin activity. These results reveal a role for the stable microtubule population in lens fiber cell elongation, acting in tandem with N-cadherin cell-cell junctions and the actomyosin network, giving insight into the cooperative role these systems play in tissue morphogenesis.

60 APPLIED LIFE SCIENCES↗

The DESI Single Fiber Lens Search. I. Four Thousand Spectroscopically Selected Galaxy–Galaxy Gravitational Lens Candidates

We present 4110 strong gravitational lens candidates, 3887 of which are new discoveries, selected from a sample of 5,837,154 luminous red galaxies (LRGs) observed with the Dark Energy Spectroscopic Instrument (DESI). Candidates are identified via the presence of background ionized oxygen [O II ] nebular emission lines in the foreground LRG spectra, which may originate from the lensing of higher-redshift star-forming galaxies. Using the measured foreground redshift, background redshift, and integrated flux of the background [O II ] doublet, we integrate over impact parameters to compute the probability that each candidate is a lens. We expect 53% of candidates to be true lenses with Einstein radii ranging from 0$^{''}_.$1–4", which can be confirmed with high-resolution imaging. Confirmed strong lenses from this sample will form a valuable cosmological data set, as strong gravitational lensing is the only method to directly measure dark matter halo substructure at cosmological distances. We independently recover the host of the multiply imaged gravitationally lensed type Ia supernova iPTF16geu. Monitoring these lenses for future multiply lensed transients will enable (a) H 0 measurements via time-delay cosmography and (b) substructure measurements via flux ratios.

Karp, Juliana S. M. [Univ. of Washington, Seattle,↗

Role of Aquaporin 0 in lens biomechanics

Maintenance of proper biomechanics of the eye lens is important for its structural integrity and for the process of accommodation to focus near and far objects. Several studies have shown that specialized cytoskeletal systems such as the beaded filament (BF) and spectrin-actin networks contribute to mammalian lens biomechanics; mutations or deletion in these proteins alters lens biomechanics. Aquaporin 0 (AQP0), which constitutes ∼45% of the total membrane proteins of lens fiber cells, has been shown to function as a water channel and a structural cell-to-cell adhesion (CTCA) protein. Our recent ex vivo study on AQP0 knockout (AQP0 KO) mouse lenses showed the CTCA function of AQP0 could be crucial for establishing the refractive index gradient. However, biomechanical studies on the role of AQP0 are lacking. The present investigation used wild type (WT), AQP5 KO (AQP5{sup −/−}), AQP0 KO (heterozygous KO: AQP0{sup +/−}; homozygous KO: AQP0{sup −/−}; all in C57BL/6J) and WT-FVB/N mouse lenses to learn more about the role of fiber cell AQPs in lens biomechanics. Electron microscopic images exhibited decreases in lens fiber cell compaction and increases in extracellular space due to deletion of even one allele of AQP0. Biomechanical assay revealed that loss of one or both alleles of AQP0 caused a significant reduction in the compressive load-bearing capacity of the lenses compared to WT lenses. Conversely, loss of AQP5 did not alter the lens load-bearing ability. Compressive load-bearing at the suture area of AQP0{sup +/−} lenses showed easy separation while WT lens suture remained intact. These data from KO mouse lenses in conjunction with previous studies on lens-specific BF proteins (CP49 and filensin) suggest that AQP0 and BF proteins could act co-operatively in establishing normal lens biomechanics. We hypothesize that AQP0, with its prolific expression at the fiber cell membrane, could provide anchorage for cytoskeletal structures like BFs and together they help to confer fiber cell shape, architecture and integrity. To our knowledge, this is the first report identifying the involvement of an aquaporin in lens biomechanics. Since accommodation is required in human lenses for proper focusing, alteration in the adhesion and/or water channel functions of AQP0 could contribute to presbyopia. - Highlights: • AQP0 aids in lens biomechanics. • AQP0 provides lens stiffness. • AQP0 is critical for lens transparency. • AQP0 could play a significant role in lens accommodation in human. • Alteration in the function(s) of lens AQP0 could lead to presbyopia.

60 APPLIED LIFE SCIENCES↗

Non-Invasive Early Detection and Molecular Analysis of Low X-ray Dose Effects in the Lens

This is the Final Progress Report for DOE-funded research project DE-PS02-08ER08-01 titled “Non-Invasive Early Detection and Molecular Analysis of Low X-ray Dose Effects in the Lens”. The project focuses on the effects of low-linear energy transfer (LET) radiation on the ocular lens. The lens is an exquisitely radiosensitive tissue with a highly-ordered molecular structure that is amenable to non-invasive optical study from the periphery. These merits point to the lens as an ideal target for laser-based molecular biodosimetry (MBD). Following exposure to different types of ionizing radiations, the lens demonstrates molecular changes (e.g., oxidation, racemization, crosslinkage, truncation, aggregation, etc.) that impact the structure and function of the long-lived proteins in the cytosol of lens fiber cells. The vast majority of proteins in the lens comprise the highly-ordered crystallins. These highly conserved lens proteins are amongst the most concentrated and stable in the body. Once synthesized, the crystallins are retained in the fiber cell cytoplasm for life. Taken together, these properties point to the lens as an ideal system for quantitative in vivo MBD assessment using quasi-elastic light scattering (QLS) analysis. In this project, we deploy a purpose-designed non-invasive infrared laser QLS instrument as a quantitative tool for longitudinal assessment of pre-cataractous molecular changes in the lenses of living mice exposed to low-dose low-LET radiation compared to non-irradiated sham controls. We hypothesize that radiation exposure will induce dose-dependent changes in the molecular structure of matrix proteins in the lens. Mechanistic assays to ascertain radiation-induced molecular changes in the lens focus on protein aggregation and gene/protein expression patterns. We anticipate that this study will contribute to our understanding of early molecular changes associated with radiation-induced tissue pathology. This study also affords potential for translational development of molecular biodosimetry instrumentation to assess human exposure to mixed radiation fields.

01 COAL, LIGNITE, AND PEAT↗

Myotonic dystrophy protein kinase (DMPK) induces actin cytoskeletal reorganization and apoptotic-like blebbing in lens cells

DMPK, the product of the DM locus, is a member of the same family of serine-threonine protein kinases as the Rho-associated enzymes. In DM, membrane inclusions accumulate in lens fiber cells producing cataracts. Overexpression of DMPK in cultured lens epithelial cells led to apoptotic-like blebbing of the plasma membrane and reorganization of the actin cytoskeleton. Enzymatically active DMPK was necessary for both effects; inactive mutant DMPK protein did not produce either effect. Active RhoA but not constitutive GDP-state mutant protein produced similar effects as DMPK. The similar actions of DMPK and RhoA suggest that they may function in the same regulatory network. The observed effects of DMPK may be relevant to the removal of membrane organelles during normal lens differentiation and the retention of intracellular membranes in DM lenses. Copyright 2000 Wiley-Liss, Inc.

Non-NASA Center↗

Integrated fiber optic probe for dynamic light scattering

An integrated fiber optic probe, comprising a monomode optical fiber fusion spliced to a short length of a graded-index multimode fiber, is fabricated for use as a coherent receiver in dynamic light scattering. The multimode fiber is cleaved to provide a gradient-index fiber lens with a focal length of 125 microns and an f-number close to unity. An integrated fiber receiver is used to measure the intensity-intensity autocorrelation data from a 0.05 percent by weight concentration of an aqueous suspension of polystyrene latex spheres. Analysis of 100 independent data sets indicates that the particle size can be recovered with an accuracy of +/- 1 percent.

Dhadwal, Harbans S.↗

946 nm Diode Pumped Laser Produces 100mJ

An innovative approach to obtaining high energy at 946 nm has yielded 101 mJ of laser energy with an optical-to-optical slope efficiency of 24.5%. A single gain module resonator was evaluated, yielding a maximum output energy of 50 mJ. In order to obtain higher energy a second gain module was incorporated into the resonator. This innovative approach produced un-surprised output energy of 101 mJ. This is of utmost importance since it demonstrates that the laser output energy scales directly with the number of gain modules. Therefore, higher energies can be realized by simply increasing the number of gain modules within the laser oscillator. The laser resonator incorporates two gain modules into a folded "M-shaped" resonator, allowing a quadruple pass gain within each rod. Each of these modules consists of a diode (stack of 30 microlensed 100 Watt diode array bars, each with its own fiber lens) end-pumping a Nd:YAG laser rod. The diode output is collected by a lens duct, which focuses the energy into a 2 mm diameter flat to flat octagonal pump area of the laser crystal. Special coatings have been developed to mitigate energy storage problems, including parasitic lasing and amplified spontaneous emission (ASE), and encourage the resonator to operate at the lower gain transition at 946 nm.

Axenson, Theresa J.↗

Apparatus for Direct Optical Fiber Through-Lens Illumination of Microscopy or Observational Objects

In one embodiment of the invention, a microscope or other observational apparatus, comprises a hollow tube, a lens mounted to the tube, a light source and at least one flexible optical fiber having an input end and an output end. The input end is positioned to receive light from the light source, and the output end is positioned within the tube so as to directly project light along a straight path to the lens to illuminate an object to be viewed. The path of projected light is uninterrupted and free of light deflecting elements. By passing the light through the lens, the light can be diffused or otherwise defocused to provide more uniform illumination across the surface of the object, increasing the quality of the image of the object seen by the viewer. The direct undeflected and uninterrupted projection of light, without change of direction, eliminates the need for light-deflecting elements, such as beam-splitters, mirrors, prisms, or the like, to direct the projected light towards the object.

Kadogawa, Hiroshi↗

Coherent fiber optic sensor for early detection of cataractogenesis in a human eye lens

A lensless backscatter fiber optic probe is used to measure the size distribution of protein molecules inside an excised, but intact, human eye lens. The fiber optic probe, about 5 mm in diameter, can be positioned arbitrarily close to the anterior surface of the eye; it is a trans-receiver, which delivers a Gaussian laser beam into a small region inside the lens and provides a coherent detection of the laser light scattered by the protein molecules in the backward direction. Protein sizes determined from the fast and slow diffusion coefficients show good correlation with the age of the lens and cataractogenesis.

Dhadwal, Harbans S.↗

High-Reliability Pump Module for Non-Planar Ring Oscillator Laser

We propose and have demonstrated a prototype high-reliability pump module for pumping a Non-Planar Ring Oscillator (NPRO) laser suitable for space missions. The pump module consists of multiple fiber-coupled single-mode laser diodes and a fiber array micro-lens array based fiber combiner. The reported Single-Mode laser diode combiner laser pump module (LPM) provides a higher normalized brightness at the combined beam than multimode laser diode based LPMs. A higher brightness from the pump source is essential for efficient NPRO laser pumping and leads to higher reliability because higher efficiency requires a lower operating power for the laser diodes, which in turn increases the reliability and lifetime of the laser diodes. Single-mode laser diodes with Fiber Bragg Grating (FBG) stabilized wavelength permit the pump module to be operated without a thermal electric cooler (TEC) and this further improves the overall reliability of the pump module. The single-mode laser diode LPM is scalable in terms of the number of pump diodes and is capable of combining hundreds of fiber-coupled laser diodes. In the proof-of-concept demonstration, an e-beam written diffractive micro lens array, a custom fiber array, commercial 808nm single mode laser diodes, and a custom NPRO laser head are used. The reliability of the proposed LPM is discussed.

micro lens array↗

Aligning Arrays of Lenses and Single-Mode Optical Fibers

A procedure now under development is intended to enable the precise alignment of sheet arrays of microscopic lenses with the end faces of a coherent bundle of as many as 1,000 single-mode optical fibers packed closely in a regular array (see Figure 1). In the original application that prompted this development, the precise assembly of lenses and optical fibers serves as a single-mode spatial filter for a visible-light nulling interferometer. The precision of alignment must be sufficient to limit any remaining wavefront error to a root-mean-square value of less than 1/10 of a wavelength of light. This wavefront-error limit translates to requirements to (1) ensure uniformity of both the lens and fiber arrays, (2) ensure that the lateral distance from the central axis of each lens and the corresponding optical fiber is no more than a fraction of a micron, (3) angularly align the lens-sheet planes and the fiber-bundle end faces to within a few arc seconds, and (4) axially align the lenses and the fiber-bundle end faces to within tens of microns of the focal distance. Figure 2 depicts the apparatus used in the alignment procedure. The beam of light from a Zygo (or equivalent) interferometer is first compressed by a ratio of 20:1 so that upon its return to the interferometer, the beam will be magnified enough to enable measurement of wavefront quality. The apparatus includes relay lenses that enable imaging of the arrays of microscopic lenses in a charge-coupled-device (CCD) camera that is part of the interferometer. One of the arrays of microscopic lenses is mounted on a 6-axis stage, in proximity to the front face of the bundle of optical fibers. The bundle is mounted on a separate stage. A mirror is attached to the back face of the bundle of optical fibers for retroreflection of light. When a microscopic lens and a fiber are aligned with each other, the affected portion of the light is reflected back by the mirror, recollimated by the microscopic lens, transmitted through the relay lenses and the beam compressor/expander, then split so that half goes to a detector and half to the interferometer. The output of the detector is used as a feedback control signal for the six-axis stage to effect alignment.

Liu, Duncan↗

A LIBS method for simultaneous monitoring of the impurities and the hydrogenic composition present in the wall of the TJ-II stellarator

The study of plasma-wall interactions and impurity transport in the plasma fusion devices is critical for the development of future fusion reactors. An experiment to perform laser induced breakdown spectroscopy, using minor modifications of our existing laser blow-off impurity injection system, has been set up thus making both experiments compatible. The radiation produced by the laser pulse focused at the TJ-II wall evaporates a surface layer of deposited impurities and the subsequent radiation produced by the laser-produced plasma is collected by two separate lens and fiber combinations into two spectrometers. The first spectrometer, with low spectral resolution, records a spectrum from 200 to 900 nm to give a survey of impurities present in the wall. The second one, with high resolution, is tuned to the wavelengths of the Hα and Dα lines in order to resolve them and quantify the hydrogen isotopic ratio present on the surface of the wall. The alignment, calibration, and spectral analysis method will be described in detail. First experimental results obtained with this setup will be shown and its relevance for the TJ-II experimental program discussed.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

A spatially resolved pyrometer for measuring the blackbody temperature of a warm dense plasma

A pyrometer has been developed to spatially resolve the blackbody temperature of a radiatively cooling warm dense plasma. The pyrometer is composed of a lens coupled fiber array, Czerny-Turner visible spectrometer, and an intensified gated CCD for the detector. The radiatively cooling warm dense plasma is generated by a ~100-ns-long intense relativistic electron bunch with an energy of 19.1 MeV and a current of 0.2 kA interacting with 100-μm-thick low-Z foils. The continuum spectrum is measured over 250 nm with a low groove density grating. These plasmas emit visible light or blackbody radiation on relatively long time scales (~0.1 to 100 μs). Finally, we presented the diagnostic layout, calibration, and proof-of-principle measurement of a radiatively cooling aluminum plasma, which includes a spatially resolved temperature gradient and the ability to temporally resolve it also.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Robotic Vision for Welding

Vision system for robotic welder looks at weld along axis of welding electrode. Gives robot view of most of weld area, including yet-unwelded joint, weld pool, and completed weld bead. Protected within welding-torch body, lens and fiber bundle give robot closeup view of weld in progress. Relayed to video camera on robot manipulator frame, weld image provides data for automatic control of robot motion and welding parameters.

Richardson, R. W.↗