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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↗

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↗

Luminescence based fiber optic probe for the detection of rare earth elements

The disclosure relates to an apparatus, method and process for detecting rare earth elements. The system includes an LED powered by a first power source and a focusing lens in optical communication with the LED. A shortpass filter is in optical communication with the focusing lens; and a fiber bifurcated cable in optical communication with the shortpass filter. The system includes a probe tip in optical communication with the fiber bifurcated cable and a sample; a first aspheric lens in optical communication with the fiber bifurcated cable. A longpass filter is in optical communication with the first aspheric lens and a second aspheric lens in optical communication with the longpass filter. The system includes a spectrometer connected to a power source, where the spectrometer is in optical communication with the second aspheric lens.

Ahern, John C.↗

Multipoint photonic doppler velocimetry using optical lens elements

A probe including a fisheye lens is disclosed to measure the velocity distribution of a moving surface along many lines of sight. Laser light, directed to the surface and then reflected back from the surface, is Doppler shifted by the moving surface, collected into fisheye lens, and then directed to detection equipment through optic fibers. The received light is mixed with reference laser light and using photonic Doppler velocimetry, a continuous time record of the surface movement is obtained. An array of single-mode optical fibers provides an optic signal to an index-matching lens and eventually to a fisheye lens. The fiber array flat polished and coupled to the index-matching lens using index-matching gel. Numerous fibers in a fiber array project numerous rays through the fisheye lens which in turn project many measurement points at numerous different locations to establish surface coverage over a hemispherical shape with very little crosstalk.

47 OTHER INSTRUMENTATION↗

Aquaporin 0 plays a pivotal role in refractive index gradient development in mammalian eye lens to prevent spherical aberration

Highlights: • Intact AQP0 functions as fiber cell-to-fiber cell adhesion protein. • AQP0 facilitates reduction in extracellular space and lens water content. • AQP0 adhesion function aids in lens refractive index gradient (RING) formation. • AQP0 prevents lens spherical aberration by establishing RING. • AQP0 is critical for lens transparency and homeostasis. - Abstract: Aquaporin 0 (AQP0) is a transmembrane channel that constitutes ∼45% of the total membrane protein of the fiber cells in mammalian lens. It is critical for lens transparency and homeostasis as mutations and knockout cause autosomal dominant lens cataract. AQP0 functions as a water channel and as a cell-to-cell adhesion (CTCA) molecule in the lens. Our recent in vitro studies showed that the CTCA function of AQP0 could be crucial to establish lens refractive index gradient (RING). However, there is a lack of in vivo data to corroborate the role of AQP0 as a fiber CTCA molecule which is critical for creating lens RING. The present investigation is undertaken to gather in vivo evidence for the involvement of AQP0 in developing lens RING. Lenses of wild type (WT) mouse, AQP0 knockout (heterozygous, AQP0{sup +/−}) and AQP0 knockout lens transgenically expressing AQP1 (heterozygous AQP0{sup +/−}/AQP1{sup +/−}) mouse models were used for the study. Data on AQP0 protein profile of intact and N- and/or C-terminal cleaved AQP0 in the lens by MALDI-TOF mass spectrometry and SDS–PAGE revealed that outer cortex fiber cells have only intact AQP0 of ∼28 kDa, inner cortical and outer nuclear fiber cells have both intact and cleaved forms, and inner nuclear fiber cells have only cleaved forms (∼26–24 kDa). Knocking out of 50% of AQP0 protein caused light scattering, spherical aberration (SA) and cataract. Restoring the lost fiber cell membrane water permeability (P{sub f}) by transgene AQP1 did not reinstate complete lens transparency and the mouse lenses showed light scattering and SA. Transmission and scanning electron micrographs of lenses of both mouse models showed increased extracellular space between fiber cells. Water content determination study showed increase in water in the lenses of these mouse models. In summary, lens transparency, CTCA and compact packing of fiber cells were affected due to the loss of 50% AQP0 leading to larger extracellular space, more water content and SA, possibly due to alteration in RING. To our knowledge, this is the first report identifying the role of AQP0 in RING development to ward off lens SA during focusing.

60 APPLIED LIFE SCIENCES↗

Near-infrared spectroscopy for divertor plasma diagnosis and control in DIII-D tokamak

New near infrared (NIR) spectroscopic measurements performed in the DIII-D tokamak divertor plasma suggest new viable diagnostic applications: divertor recycling and low-Z impurity flux measurements, a spectral survey for divertor Thomson scattering (DTS) diagnostic, and T{sub e} monitoring for divertor detachment control. A commercial 0.3 m spectrometer coupled to an imaging lens via optical fiber and a InGaAs 1024 pixel array detector enabled deuterium and impurity emission measurements in the range 800–2300 nm. The first full NIR survey identified D, He, B, Li, C, N, O, Ne lines and provided plasma T{sub e}, n{sub e} estimates from deuterium Paschen and Brackett series intensity and Stark line broadening analysis. The range 1.000–1.060 mm was surveyed in high-density and neon seeded divertor plasmas for spectral background emission studies for λ = 1.064 μm laser-based DTS development. The ratio of adjacent deuterium Paschen-α and Brackett Br9 lines in recombining divertor plasmas is studied for divertor T{sub e} monitoring aimed at divertor detachment real-time feedback control.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

SU-E-T-20: A Novel Hybrid CBCT, Bioluminescence and Fluorescence Tomography System for Preclinical Radiation Research

Purpose: A novel standalone bioluminescence and fluorescence tomography (BLT and FT) system equipped with high resolution CBCT has been built in our group. In this work, we present the system calibration method and validate our system in both phantom and in vivo environment. Methods: The CBCT is acquired by rotating the animal stage while keeping the x-ray source and detector panel static. The optical signal is reflected by the 3-mirror system to a multispectral filter set and then delivered to the CCD camera with f/1.4 lens mounted. Nine fibers passing through the stage and in contact with the mouse skin serve as the light sources for diffuse optical tomography (DOT) and FT. The anatomical information and optical properties acquired from the CBCT and DOT, respectively, are used as the priori information to improve the BLT/FT reconstruction accuracy. Flat field correction for the optical system was acquired at multiple wavelengths. A home-built phantom is used to register the optical and CBCT coordinates. An absolute calibration relating the CCD photon counts rate to the light fluence rate emitted at animal surface was developed to quantify the bioluminescence power or fluorophore concentration. Results: An optical inhomogeneous phantom with 2 light sources (3mm separation) imbedded is used to test the system. The optical signal is mapped onto the mesh generated from CBCT for optical reconstruction. Our preliminary results show that the center of mass can be reconstructed within 2.8mm accuracy. A live mouse with the light source imbedded is also used to validate our system. Liver or lung metastatic luminescence tumor model will be used for further testing. Conclusion: This hybrid system transforms preclinical research to a level that even sub-palpable volume of cells can be imaged rapidly and non-invasively, which largely extends the scope of radiobiological research. The research is supported by the NCI grant R01CA158100-01.

60 APPLIED LIFE SCIENCES↗

Systematic evaluation of photodetector performance for plastic scintillation dosimetry

Purpose: The authors’ objective was to systematically assess the performance of seven photodetectors used in plastic scintillation dosimetry. The authors also propose some guidelines for selecting an appropriate detector for a specific application. Methods: The plastic scintillation detector (PSD) consisted of a 1-mm diameter, 10-mm long plastic scintillation fiber (BCF-60), which was optically coupled to a clear 10-m long optical fiber of the same diameter. A light-tight plastic sheath covered both fibers and the scintillator end was sealed. The clear fiber end was connected to one of the following photodetectors: two polychromatic cameras (one with an optical lens and one with a fiber optic taper replacing the lens), a monochromatic camera with an optical lens, a PIN photodiode, an avalanche photodiode (APD), or a photomultiplier tube (PMT). A commercially available W1 PSD was also included in the study, but it relied on its own fiber and scintillator. Each PSD was exposed to both low-energy beams (120, 180, and 220 kVp) from an orthovoltage unit and high-energy beams (6 and 23 MV) from a linear accelerator. Various dose rates were tested to identify the operating range and accuracy of each photodetector. Results: For all photodetectors, the relative uncertainty was less than 5% for dose rates higher than 3 mGy/s. The cameras allowed multiple probes to be used simultaneously, but they are less sensitive to low-light signals. The PIN, APD, and PMT had higher sensitivity, making them more suitable for low dose rate and out-of-field dose monitoring. The relative uncertainty of the PMT was less than 1% at the lowest dose rate achieved (0.10 mGy/s), suggesting that it was optimal for use in live dosimetry. Conclusions: For dose rates higher than 3 mGy/s, the PIN diode is the most effective photodetector in terms of performance/cost ratio. For lower dose rates, such as those seen in interventional radiology or high-gradient radiotherapy, PMTs are the optimal choice.

60 APPLIED LIFE SCIENCES↗

TH-C-19A-10: Systematic Evaluation of Photodetectors Performances for Plastic Scintillation Dosimetry

Purpose: To assess and compare the performance of different photodetectors likely to be used in a plastic scintillation detector (PSD). Methods: The PSD consists of a 1 mm diameter, 10 mm long plastic scintillation fiber (BCF-60) which is optically coupled to a clear 10 m long optical fiber of the same diameter. A light-tight plastic sheath covers both fibers and the scintillator end is sealed. The clear fiber end is connected to one of the following six studied photodetectors: two polychromatic cameras (one with an optical lens and one with a fiber optic taper replacing the lens); a monochromatic camera with the same optical lens; a PIN photodiode; an avalanche photodiode (APD); and a photomultiplier tube (PMT). Each PSD is exposed to both low energy beams (120, 180, and 220 kVp) from an orthovoltage unit, and high energy beams (6 MV and 23 MV) from a linear accelerator. Various dose rates are explored to identify the photodetectors operating ranges and accuracy. Results: For all photodetectors, the relative uncertainty remains under 5 % for dose rates over 3 mGy/s. The taper camera collects four times more signal than the optical lens camera, although its standard deviation is higher since it could not be cooled. The PIN, APD and PMT have higher sensitivity, suitable for low dose rate and out-of-field dose monitoring. PMT's relative uncertainty remains under 1 % at the lowest dose rate achievable (50 μGy/s), suggesting optimal use for live dosimetry. Conclusion: A set of 6 photodetectors have been studied over a broad dose rate range at various energies. For dose rate above 3 mGy/s, the PIN diode is the most effective photodetector in term of performance/cost ratio. For lower dose rate, such as those seen in interventional radiology, PMTs are the optimal choice. FQRNT Doctoral Research Scholarship.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Photonic Doppler velocimetry probe used to measure grain boundaries of dynamic shocked materials

Material scientists have developed computational modeling to predict the dynamic response of materials undergoing stress, but there is still a need to make precision measurements of surfaces undergoing shock compression. Miniature photonic Doppler velocimetry (PDV) probes have been developed to measure the velocity distribution from a moving surface traveling tens of kilometers per second. These probes use hundreds of optical fibers imaged by optical relays onto different regions of this moving surface. While previous work examined large surface areas, we have now developed a PDV microscope that can interrogate 37 different spots within a field of view of <1 mm, with a standoff distance of 17 mm, to analyze the motion differences across grain boundaries of the material undergoing dynamic stress. Each PDV fiber interrogates a 10 μm spot size on the moving surface. A separate imaging system using a coherent bundle records the location of the PDV spots relative to the grain boundaries prior to the dynamic event. Designing the mounting structures for the lenses, fibers, and coherent bundle was a challenge. To minimize back reflections, the fibers are index matched onto the first relay lens, which is made of fused silica. The PDV fibers are aligned normal to the moving surface. The imaging probe views the surface at an 18° angle. The coherent bundle is tilted 11° to its optical relay. All components are assembled into a single probe head assembly. The coherent bundle is removed from the probe head to be used for the next dynamic event. Alignment issues will also be discussed.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Optical integration of Pancharatnam-Berry phase lens and dynamical phase lens

In the optical system, most elements such as lens, prism, and optical fiber are made of silica glass. Therefore, integrating Pancharatnam-Berry phase elements into silica glass has potential applications in the optical system. In this paper, we take a lens, for example, which integrates a Pancharatnam-Berry phase lens into a conventional plano-convex lens. The spin states and positions of focal points can be modulated by controlling the polarization states of the incident beam. The proposed lens has a high transmission efficiency, and thereby acts as a simple and powerful tool to manipulate spin photons. Furthermore, the method can be conveniently extended to the optical fiber and laser cavity, and may provide a route to the design of the spin-photonic devices.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗