Developing retinal cell therapy: cones and cone-like cells in transplantation and development

Retinal cell replacement therapy aims to restore vision in retinal degenerative diseases by replacing dead photoreceptors. Injecting GFP+ rod photoreceptor precursors into the subretinal space of recipient mice leads to GFP+ cells being seen in the recipient retina. In models of retinal degeneration, replacement of proteins lacking in the recipient, as well as functional improvement, has been seen. This thesis aimed to extend this work to use cone photoreceptors, which are more important for human vision. The Chrnb4.eGFP model was selected as a source of cones, as well as the Nrl-/- and Nr2e3rd7/rd7 models which generate an increased number of cone-like cells. Cells were injected into the subretinal space of several different mouse recipients, and a number of retinal degenerative models representing a range of cone to rod ratios and functionalities. GFP+ photoreceptors, including unambiguous morphology and immunohistochemical markers were seen in recipient retinas after transplantation into all tested recipient types. The highest numbers occurred in Nrl-deficient and Prph2 mutant mice. The majority of GFP+ cells resembled rod photoreceptors in morphology, and did not express cone-specific markers, except in Nrl-deficient recipients. Evidence from these and other experiments showed that these results were most likely due not to cell integration but instead to the transfer of material including GFP from injected cells to existing recipient photoreceptors. To investigate functional outcomes of transplantation, electroretinography and multi-electrode array (MEA) techniques were used. MEA data showed no clear improvement in light response in treated retinas. Time-lapse imaging of explanted early postnatal retinal tissue using multi-photon microscopy was used to investigate the migratory behaviour of developing cone photoreceptor precursors around the ages of transplantation. This revealed a cyclical pattern of migration similar to interkinetic nuclear migration, with slow basal and rapid apical movements. Pharmacological intervention implicated the dynein/kinesin motor proteins in the apical movement seen

Repeated nuclear translocations underlie photoreceptor positioning and lamination of the outer nuclear layer in the mammalian retina

In development, almost all stratified neurons must migrate from their birthplace to the appropriate neural layer. Photoreceptors reside in the most apical layer of the retina, near their place of birth. Whether photoreceptors require migratory events for fine-positioning and/or retention within this layer is not well understood. Here, we show that photoreceptor nuclei of the developing mouse retina cyclically exhibit rapid, dynein-1-dependent translocation toward the apical surface, before moving more slowly in the basal direction, likely due to passive displacement by neighboring retinal nuclei. Attenuating dynein 1 function in rod photoreceptors results in their ectopic basal displacement into the outer plexiform layer and inner nuclear layer. Synapse formation is also compromised in these displaced cells. We propose that repeated, apically directed nuclear translocation events are necessary to ensure retention of post-mitotic photoreceptors within the emerging outer nuclear layer during retinogenesis, which is critical for correct neuronal lamination

Assessing human diet and movement in the Tongan maritime chiefdom using isotopic analyses.

The rise of stratified societies fundamentally influences the interactions between status, movement, and food. Using isotopic analyses, we assess differences in diet and mobility of individuals excavated from two burial mounds located at the `Atele burial site on Tongatapu, the main island of the Kingdom of Tonga (c. 500 - 150 BP). The first burial mound (To-At-1) was classified by some archaeologists as a commoner's mound while the second burial mound (To-At-2) was possibly used for interment of the chiefly class. In this study, stable isotope analyses of diet (δ13C, δ15N, and δ34S; n = 41) are used to asses paleodiet and 87Sr/86Sr ratios (n = 30) are analyzed to investigate individual mobility to test whether sex and social status affected these aspects of life. Our results show significant differences in diet between burial mounds and sexes. Those interred in To-At-2 displayed lower δ13C values, indicating they ate relatively more terrestrial plants (likely starchy vegetable staples) compared with To-At-1 individuals. Females displayed significantly lower δ15N values compared with males within the entire assemblage. No differences in δ34S values were observed between sexes or burial mound but it is possible that sea spray or volcanism may have affected these values. One individual displayed the strontium isotopic composition representative of a nonlocal immigrant (outside 2SD of the mean). This suggests the hegemonic control over interisland travel, may have prevented long-term access to the island by non-Tongans exemplifying the political and spiritual importance of the island of Tongatapu in the maritime chiefdom

An Antimicrobial Peptide Regulates Tumor-Associated Macrophage Trafficking via the Chemokine Receptor CCR2, a Model for Tumorigenesis

Tumor-associated macrophages (TAMs) constitute a significant part of infiltrating inflammatory cells that are frequently correlated with progression and poor prognosis of a variety of cancers. Tumor cell-produced human β-defensin-3 (hBD-3) has been associated with TAM trafficking in oral cancer; however, its involvement in tumor-related inflammatory processes remains largely unknown., applying a cross-desensitization strategy of CCR2 and its pharmacological inhibitor (RS102895), respectively, was also carried out. outcome and demonstrates the importance of the innate immune system in the development of tumors

The Long-Baseline Neutrino Experiment: Exploring Fundamental Symmetries of the Universe

Major update of previous version. This is the reference document for LBNE science program and current status. Chapters 1, 3, and 9 provide a comprehensive overview of LBNE's scientific objectives, its place in the landscape of neutrino physics worldwide, the technologies it will incorporate and the capabilities it will possess. 288 pages, 116 figuresMajor update of previous version. This is the reference document for LBNE science program and current status. Chapters 1, 3, and 9 provide a comprehensive overview of LBNE's scientific objectives, its place in the landscape of neutrino physics worldwide, the technologies it will incorporate and the capabilities it will possess. 288 pages, 116 figuresThe preponderance of matter over antimatter in the early Universe, the dynamics of the supernova bursts that produced the heavy elements necessary for life and whether protons eventually decay --- these mysteries at the forefront of particle physics and astrophysics are key to understanding the early evolution of our Universe, its current state and its eventual fate. The Long-Baseline Neutrino Experiment (LBNE) represents an extensively developed plan for a world-class experiment dedicated to addressing these questions. LBNE is conceived around three central components: (1) a new, high-intensity neutrino source generated from a megawatt-class proton accelerator at Fermi National Accelerator Laboratory, (2) a near neutrino detector just downstream of the source, and (3) a massive liquid argon time-projection chamber deployed as a far detector deep underground at the Sanford Underground Research Facility. This facility, located at the site of the former Homestake Mine in Lead, South Dakota, is approximately 1,300 km from the neutrino source at Fermilab -- a distance (baseline) that delivers optimal sensitivity to neutrino charge-parity symmetry violation and mass ordering effects. This ambitious yet cost-effective design incorporates scalability and flexibility and can accommodate a variety of upgrades and contributions. With its exceptional combination of experimental configuration, technical capabilities, and potential for transformative discoveries, LBNE promises to be a vital facility for the field of particle physics worldwide, providing physicists from around the globe with opportunities to collaborate in a twenty to thirty year program of exciting science. In this document we provide a comprehensive overview of LBNE's scientific objectives, its place in the landscape of neutrino physics worldwide, the technologies it will incorporate and the capabilities it will possess

Demonstration of a high repetition rate capillary discharge waveguide

A hydrogen-filled capillary discharge waveguide operating at kHz repetition rates is presented for parameters relevant to laser plasma acceleration (LPA). The discharge current pulse was optimized for erosion mitigation with laser guiding experiments and MHD simulation. Heat flow simulations and measurements showed modest temperature rise at the capillary wall due to the average heat load at kHz repetition rates with water-cooled capillaries, which is promising for applications of LPAs such as high average power radiation sources

Mandibular morphology as an indicator of human subadult age: Interlandmark approaches

The dentition is widely recognized as the set of developmental markers that appear to show the least variability against chronological age; these markers are thus widely used in forensic anthropological investigations. As a possible alternative, we investigate here the potential of mandibular morphology as a developmental marker for estimating age at death in subadults. The sample analyzed comprises 79 known age and sex subadult individuals of South African Bantu and African American origin. Linear measurements of ramus height were obtained from the mathematical conversion of three-dimensional landmark data. A series of regression analyses were then performed to predict age by using the measurement of ramus height; results were cross-validated using a jackknife procedure. Our results show that ramus height can be used to predict age in the subadult skeleton with accuracy, closely approaching that of standards based on the dentition (standard error rates are between +/- 1.1 years and +/- 2.4 years)