Ocular Gene Therapy with Adeno-associated Virus Vectors: Current Outlook for Patients and Researchers

In this “Perspective”, we discuss ocular gene therapy – the patient’s perspective, the various strategies of gene replacement and gene editing, the place of adenoassociated virus vectors, routes of delivery to the eye and the remaining question - “why does immunity continue to limit efficacy?” Through the coordinated efforts of patients, researchers, granting agencies and industry, and after many years of pre-clinical studies, biochemical, cellular, and animal models, we are seeing clinical trials emerge for many previously untreatable heritable ocular disorders. The pathway to therapies has been led by the successful treatment of the RPE65 form of Leber congenital amaurosis with LUXTURNATM. In some cases, immune reactions to the vectors continue to occur, limiting efficacy. The underlying mechanisms of inflammation require further study, and new vectors need to be designed that limit the triggers of immunity. Researchers studying ocular gene therapies and clinicians enrolling patients in clinical trials must recognize the current limitations of these therapies to properly manage expectations and avoid disappointment, but we believe that gene therapies are well on their way to successful, widespread utilization to treat heritable ocular disorders

Recent pace of change in human impact on the world's ocean.

Humans interact with the oceans in diverse and profound ways. The scope, magnitude, footprint and ultimate cumulative impacts of human activities can threaten ocean ecosystems and have changed over time, resulting in new challenges and threats to marine ecosystems. A fundamental gap in understanding how humanity is affecting the oceans is our limited knowledge about the pace of change in cumulative impact on ocean ecosystems from expanding human activities - and the patterns, locations and drivers of most significant change. To help address this, we combined high resolution, annual data on the intensity of 14 human stressors and their impact on 21 marine ecosystems over 11 years (2003-2013) to assess pace of change in cumulative impacts on global oceans, where and how much that pace differs across the ocean, and which stressors and their impacts contribute most to those changes. We found that most of the ocean (59%) is experiencing significantly increasing cumulative impact, in particular due to climate change but also from fishing, land-based pollution and shipping. Nearly all countries saw increases in cumulative impacts in their coastal waters, as did all ecosystems, with coral reefs, seagrasses and mangroves at most risk. Mitigation of stressors most contributing to increases in overall cumulative impacts is urgently needed to sustain healthy oceans

Genetic protocols for DNA extraction from white-tailed deer cast antlers to confirm individuality

White-tailed deer (Odocoileus virginianus) are the most sought-after deer species in America. The antlers of mammals, such as deer, are one of the fastest regenerative tissues in the world and are grown and naturally cast every year. Research on cast antlers have been used for a variety of purposes including population comparisons and impacts of deer health due to climatic stressors. When investigating cast antlers, it is important to confirm individuality of match sets in addition to antlers of the same individual between years. Therefore, individuality must be confirmed genetically, and protocols must be developed and established to do so. Our objectives were to 1) establish a genetic protocol to harvest DNA from cast antlers using connective tissue, and 2) determine individuality from subsequent years and match sets. When fresh antlers are cast, they leave behind a viable connective tissue from which DNA can be extracted. The DNA was successfully extracted from the skin rings harvested from naturally cast antlers. This study developed viable methods to confirm individuality, which aid researchers and wildlife biologists in a better understanding of the white-tailed deer herd as they set management goals and harvest regulations

Human Pluripotent Stem Cells Serve as an Effective In Vitro Model for Studies of Early Stages of Retinogenesis

poster abstractSpecification of the neural retina is one of the first events in human development and thus, efforts to study the initial stages of retinal specification have been largely limited. This is particularly true for the earliest event in retinogenesis, the establishment of a definitive retinal fate from a more primitive neural progenitor source. With the advent of human pluripotent stem cells (hPSCs), the complex interplay of transcription factors involved in early events of retinal development from an unspecified pluripotent population can be studied in an in vitro model. To examine this potential, hPSCs were directed to differentiate to a neuroretinal lineage in which a subpopulation of cells adopts a definitive retinal fate whereas others develop toward a forebrain lineage. Samples were collected over the first month of differentiation, starting from the undifferentiated state through when cells acquired either retinal or nonretinal forebrain identities and gene expression patterns were characterized using immunocytochemistry and quantitative RT-PCR. Results demonstrated that while neural transcription factors such as PAX6, OTX2, and LHX2 were expressed early in development, definite retinal transcription factors such as CHX10 were expressed later in differentiation. Furthermore, the expression of CHX10 was found to be uniquely associated with retinal populations and remained absent from the other neural populations, thereby illustrating the ability of this protocol to uniquely identify and isolate retinal populations and further study them in vitro. Overall, these studies will serve to further understand the specification of a retinal fate from a pluripotent population. Such information will assist in the establishment of more efficient methods to generate retinal cells from hiPSCs for translational purposes, as well as establish hiPSCs as a unique in vitro model system for studies of the earliest stages of human development. Mentor: Jason S. Meyer1-

The spectral and chemical measurement of pollutants on snow near South Pole, Antarctica

Remote sensing of light-absorbing particles (LAPs), or dark colored impurities, such as black carbon (BC) and dust on snow, is a key remaining challenge in cryospheric surface characterization and application to snow, ice, and climate models. We present a quantitative data set of in situ snow reflectance, measured and modeled albedo, and BC and trace element concentrations from clean to heavily fossil fuel emission contaminated snow near South Pole, Antarctica. Over 380 snow reflectance spectra (350–2500 nm) and 28 surface snow samples were collected at seven distinct sites in the austral summer season of 2014–2015. Snow samples were analyzed for BC concentration via a single particle soot photometer and for trace element concentration via an inductively coupled plasma mass spectrometer. Snow impurity concentrations ranged from 0.14 to 7000 part per billion (ppb) BC, 9.5 to 1200 ppb sulfur, 0.19 to 660 ppb iron, 0.013 to 1.9 ppb chromium, 0.13 to 120 ppb copper, 0.63 to 6.3 ppb zinc, 0.45 to 82 parts per trillion (ppt) arsenic, 0.0028 to 6.1 ppb cadmium, 0.062 to 22 ppb barium, and 0.0044 to 6.2 ppb lead. Broadband visible to shortwave infrared albedo ranged from 0.85 in pristine snow to 0.62 in contaminated snow. LAP radiative forcing, the enhanced surface absorption due to BC and trace elements, spanned from \u3c1 W m­–2 for clean snow to ~70 W m­–2 for snow with high BC and trace element content. Measured snow reflectance differed from modeled snow albedo due to specific impurity-dependent absorption features, which we recommend be further studied and improved in snow albedo models

Comparison of Satellite-Derived and In-Situ Observations of Ice and Snow Surface Temperatures over Greenland

The most practical way to get a spatially broad and continuous measurements of the surface temperature in the data-sparse cryosphere is by satellite remote sensing. The uncertainties in satellite-derived LSTs must be understood to develop internally-consistent decade-scale land-surface temperature (LST) records needed for climate studies. In this work we assess satellite-derived "clear-sky" LST products from the Moderate Resolution Imaging Spectroradiometer (MODIS) and the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER), and LSTs derived from the Enhanced Thematic Mapper Plus (ETM+) over snow and ice on Greenland. When possible, we compare satellite-derived LSTs with in-situ air-temperature observations from Greenland Climate Network (GC-Net) automatic-weather stations (AWS). We find that MODIS, ASTER and ETM+ provide reliable and consistent LSTs under clear-sky conditions and relatively-flat terrain over snow and ice targets over a range of temperatures from -40 to 0 C. The satellite-derived LSTs agree within a relative RMS uncertainty of approx.0.5 C. The good agreement among the LSTs derived from the various satellite instruments is especially notable since different spectral channels and different retrieval algorithms are used to calculate LST from the raw satellite data. The AWS record in-situ data at a "point" while the satellite instruments record data over an area varying in size from: 57 X 57 m (ETM+), 90 X 90 m (ASTER), or to 1 X 1 km (MODIS). Surface topography and other factors contribute to variability of LST within a pixel, thus the AWS measurements may not be representative of the LST of the pixel. Without more information on the local spatial patterns of LST, the AWS LST cannot be considered valid ground truth for the satellite measurements, with RMS uncertainty approx.2 C. Despite the relatively large AWS-derived uncertainty, we find LST data are characterized by high accuracy but have uncertain absolute precision

Early Spring Post-Fire Snow Albedo Dynamics in High Latitude Boreal Forests Using Landsat-8 OLI Data

Taking advantage of the improved radiometric resolution of Landsat-8 OLI which, unlike previous Landsat sensors, does not saturate over snow, the progress of fire recovery progress at the landscape scale (less than 100 m) is examined. High quality Landsat-8 albedo retrievals can now capture the true reflective and layered character of snow cover over a full range of land surface conditions and vegetation densities. This new capability particularly improves the assessment of post-fire vegetation dynamics across low- to high-burn severity gradients in Arctic and boreal regions in the early spring, when the albedos during recovery show the greatest variation. We use 30 m resolution Landsat-8 surface reflectances with concurrent coarser resolution (500 m) MODIS high quality full inversion surface Bidirectional Reflectance Distribution Functions (BRDF) products to produce higher resolution values of surface albedo. The high resolution full expression shortwave blue sky albedo product performs well with an overall RMSE of 0.0267 between tower and satellite measures under both snow-free and snow-covered conditions. While the importance of post-fire albedo recovery can be discerned from the MODIS albedo product at regional and global scales, our study addresses the particular importance of early spring post-fire albedo recovery at the landscape scale by considering the significant spatial heterogeneity of burn severity, and the impact of snow on the early spring albedo of various vegetation recovery types. We found that variations in early spring albedo within a single MODIS gridded pixel can be larger than 0.6. Since the frequency and severity of wildfires in Arctic and boreal systems is expected to increase in the coming decades, the dynamics of albedo in response to these rapid surface changes will increasingly impact the energy balance and contribute to other climate processes and physical feedback mechanisms. Surface radiation products derived from Landsat-8 data will thus play an important role in characterizing the carbon cycle and ecosystem processes of high latitude systems

A Blended Global Snow Product using Visible, Passive Microwave and Scatterometer Satellite Data

A joint U.S. Air Force/NASA blended, global snow product that utilizes Earth Observation System (EOS) Moderate Resolution Imaging Spectroradiometer (MODIS), Advanced Microwave Scanning Radiometer for EOS (AMSR-E) and QuikSCAT (Quick Scatterometer) (QSCAT) data has been developed. Existing snow products derived from these sensors have been blended into a single, global, daily, user-friendly product by employing a newly-developed Air Force Weather Agency (AFWA)/National Aeronautics and Space Administration (NASA) Snow Algorithm (ANSA). This initial blended-snow product uses minimal modeling to expeditiously yield improved snow products, which include snow cover extent, fractional snow cover, snow water equivalent (SWE), onset of snowmelt, and identification of actively melting snow cover. The blended snow products are currently 25-km resolution. These products are validated with data from the lower Great Lakes region of the U.S., from Colorado during the Cold Lands Processes Experiment (CLPX), and from Finland. The AMSR-E product is especially useful in detecting snow through clouds; however, passive microwave data miss snow in those regions where the snow cover is thin, along the margins of the continental snowline, and on the lee side of the Rocky Mountains, for instance. In these regions, the MODIS product can map shallow snow cover under cloud-free conditions. The confidence for mapping snow cover extent is greater with the MODIS product than with the microwave product when cloud-free MODIS observations are available. Therefore, the MODIS product is used as the default for detecting snow cover. The passive microwave product is used as the default only in those areas where MODIS data are not applicable due to the presence of clouds and darkness. The AMSR-E snow product is used in association with the difference between ascending and descending satellite passes or Diurnal Amplitude Variations (DAV) to detect the onset of melt, and a QSCAT product will be used to map areas of snow that are actively melting

Neural correlates of response inhibition in early childhood: Evidence from a Go/No-go task

We examined the neural correlates underlying response inhibition in early childhood. Fiveyear-old children completed a Go/No-go task with or without time pressure (Fast vs. Slow condition) while scalp EEG was recorded. On No-go trials where inhibition was required, the left frontal N2 and posterior P3 were enhanced relative to Go trials. Time pressure was detrimental to behavioural performance and modulated the early-occurring P1 component. The topography of ERPs related to response inhibition differed from patterns typically seen in adults, and may indicate a compensatory mechanism to make up for immature inhibition networks in children