Homology Cloning, Heterologous Expression and Characterization of a New Channelrhodopsin

Channelrhodopsins are phototaxis receptors in the plasma membranes of motile unicellular algae. They function as light-gated cation channels and this channel activity has been exploited to trigger action potentials in neurons with light to control neural circuits (“optogenetics"). Four channelrhodopsins were identified in two algal species, Chlamydomonas reinhardtii and Volvox carteri, with known genome sequences; each species contains 2 channelrhodopsins, one absorbing at longer wavelengths and one at shorter wavelengths, named CrChR1 and CrChR2, respectively. Our goals are to expand knowledge of channelrhodopsin mechanisms and also to identify new channelrhodopsins from various algal species with improved properties for optogenetic use. For these aims we are targeting algae from extreme environments to establish the natural diversity of their properties. We cloned a new channelrhodopsin from the psychrophilic (cold-loving) alga, Chlamydomonas augustae, with degenerate primers based on the 4 known homologs. The new protein is 48% and 52% identical to CrChR1 and CrChR2, respectively. We expressed the channelrhodopsin in HEK293 cells and measured light-induced currents to assess their kinetics and action spectrum. Based on the primary structure, kinetics of light-induced photocurrents in HEK293 cells, and action spectrum maximum of 520 nm near that of the two previously found CrChR1, we named the new channelrhodopsin CaChR1. The properties of robust channel activity at physiological pH, fast on-and-off kinetics, and greatly red-shifted action spectrum maximum from that of CrChR2, make CaChR1 advantageous as an optogenetic tool. To know this new channelrhodopsin better, we expressed His-tagged CaChR1 in Pichia pastoris and the yield is about 6 mg/L. The purified His-tagged CaChR1 exhibited an absorption spectrum identical to the action spectrum of CaChR1-generated photocurrents. The future work will be measurement of the photocycles of CaChR1 by flash photolysis, crystallization of CaChR1 for the structure and mutagenesis of CaChR1 to find the critical amino acids accounting for red-shifted spectra, slow inactivation and rapid on-and-off kinetics. Seven new channelrhodopsins including CaChR1 from different algal species have been cloned in our lab at this time, bringing the total known to 13. The work of cloning of these new channelrhodopsins along with the expression of CaChR1 was published in Photochemistry and Photobiology in January 2012

Development of a SPECT -based three-dimensional treatment planner for radionuclide therapy with iodine -131

Accurate calculation of absorbed dose to target tumors and normal tissues in the body is an important requirement for establishing fundamental dose-response relationships for radioimmunotherapy. Two major obstacles have been the difficulty in obtaining an accurate patient-specific 3-D activity map in-vivo and calculating the resulting absorbed dose. This study investigated a methodology for 3-D internal dosimetry, which integrates the 3-D biodistribution of the radionuclide acquired from SPECT with a dose-point kernel convolution technique to provide the 3-D distribution of absorbed dose. Accurate SPECT images were reconstructed with appropriate methods for noise filtering, attenuation correction, and Compton scatter correction. The SPECT images were converted into activity maps using a calibration phantom. The activity map was convolved with an $\sp{131}$I dose-point kernel using a 3-D fast Fourier transform to yield a 3-D distribution of absorbed dose. The 3-D absorbed dose map was then processed to provide the absorbed dose distribution in regions of interest. This methodology can provide heterogeneous distributions of absorbed dose in volumes of any size and shape with nonuniform distributions of activity. Comparison of the activities quantitated by our SPECT methodology to true activities in an Alderson abdominal phantom (with spleen, liver, and spherical tumor) yielded errors of $-$16.3% to 4.4%. Volume quantitation errors ranged from $-$4.0 to 5.9% for volumes greater than 88 ml. The percentage differences of the average absorbed dose rates calculated by this methodology and the MIRD S-values were 9.1% for liver, 13.7% for spleen, and 0.9% for the tumor. Good agreement (percent differences were less than 8%) was found between the absorbed dose due to penetrating radiation calculated from this methodology and TLD measurement. More accurate estimates of the 3-D distribution of absorbed dose can be used as a guide in specifying the minimum activity to be administered to patients to deliver a prescribed absorbed dose to tumor without exceeding the toxicity limits of normal tissues. ^

Alpine permafrost index map

The objective of this study is the production of an Alpine Permafrost Index Map (APIM) covering the entire European Alps. A unified statistical model that is based on Alpine-wide permafrost observations is used for debris and bedrock surfaces across the entire Alps. The explanatory variables of the model are mean annual air temperatures, potential incoming solar radiation and precipitation. Offset terms were applied to make model predictions for topographic and geomorphic conditions that differ from the terrain features used for model fitting. These offsets are based on literature review and involve some degree of subjective choice during model building. The assessment of the APIM is challenging because limited independent test data are available for comparison and these observations represent point information in a spatially highly variable topography. The APIM provides an index that describes the spatial distribution of permafrost and comes together with an interpretation key that helps to assess map uncertainties and to relate map contents to their actual expression in terrain. The map can be used as a first resource to estimate permafrost conditions at any given location in the European Alps in a variety of contexts such as research and spatial planning. Results show that Switzerland likely is the country with the largest permafrost area in the Alps, followed by Italy, Austria, France and Germany. Slovenia and Liechtenstein may have marginal permafrost areas. In all countries the permafrost area is expected to be larger than the glacier-covered area.

Reconstructed glaciation limits in Eurasia (digitized from map Fig.1)

Based on field investigations in northern Russia and interpretation of offshore seismic data, we have made a preliminary reconstruction of the maximum ice-sheet extent in the Barents and Kara Sea region during the Early/Middle Weichselian and the Late Weichselian. Our investigations indicate that the Barents and Kara ice sheets attained their maximum Weichselian positions in northern Russia prior to 50 000 yr BP, whereas the northeastern flank of the Scandinavian Ice Sheet advanced to a maximum position shortly after 17000 calendar years ago. During the Late Weichselian (25 000-10000 yr BP), much of the Russian Arctic remained ice-free. According to our reconstruction, the extent of the ice sheets in the Barents and Kara Sea region during the Late Weichselian glacial maximum was less than half that of the maximum model which, up to now, has been widely used as a boundary condition for testing and refining General Circulation Models (GCMs). Preliminary numerical-modelling experiments predict Late Weichselian ice sheets which are larger than the ice extent implied for the Kara Sea region from dated geological evidence, suggesting very low precipitation.

Map of Lewis-Glacier - Mt. Kenya 1983 (pdf 600 kB)

In February of 1983 a new terrestrial photogrammetric survey of Lewis Glacier (0° 9' S) has been made, from which the present topographic map has been produced in a scale of 1:5000. Simultaneously a survey of 1963 was evaluated giving a basis for computations of area and volume changes over the 20 year period: Lewis Glacier has lost 22 % of its area and 50 % of its volume. Based on maps and field observations of moraines 10 different stages were identified. Changes of area and volume can be determined for the periods after 1890, two older, undated stages are presumed to be of Little Ice Age-origin. Moderate losses from 1890 to 1920 were followed by strong, uninterrupted retreat up to present. In this respect Lewis Glacier behaves as all other equatorial glaciers that were closer examined. Compared to alpine glaciers the development was similar up to 1950. In the following years, however, the glaciers of the Alps gained mass and advanced while Lewis Glacier experienced its strongest losses from 1974 to 1983.

Code and readme of a MATLAB-based graphical user interface program for computing functionals of the geopotential

We present a novel graphical user interface program GrafLab (GRAvity Field LABoratory) for spherical harmonic synthesis (SHS) created in MATLAB®. This program allows to comfortably compute 38 various functionals of the geopotential up to ultra-high degrees and orders of spherical harmonic expansion. For the most difficult part of the SHS, namely the evaluation of the fully normalized associated Legendre functions (fnALFs), we used three different approaches according to required maximum degree: (i) the standard forward column method (up to maximum degree 1800, in some cases up to degree 2190); (ii) the modified forward column method combined with Horner's scheme (up to maximum degree 2700); (iii) the extended-range arithmetic (up to an arbitrary maximum degree). For the maximum degree 2190, the SHS with fnALFs evaluated using the extended-range arithmetic approach takes only approximately 2-3 times longer than its standard arithmetic counterpart, i.e. the standard forward column method. In the GrafLab, the functionals of the geopotential can be evaluated on a regular grid or point-wise, while the input coordinates can either be read from a data file or entered manually. For the computation on a regular grid we decided to apply the lumped coefficients approach due to significant time-efficiency of this method. Furthermore, if a full variance-covariance matrix of spherical harmonic coefficients is available, it is possible to compute the commission errors of the functionals. When computing on a regular grid, the output functionals or their commission errors may be depicted on a map using automatically selected cartographic projection.

Plant Functional Type (PFT) map over Siberia derived from GlobCover 2005 dataset, link to dataset in NetCDF format

High-latitude ecosystems play an important role in the global carbon cycle and in regulating the climate system and are presently undergoing rapid environmental change. Accurate land cover data sets are required to both document these changes as well as to provide land-surface information for benchmarking and initializing Earth system models. Earth system models also require specific land cover classification systems based on plant functional types (PFTs), rather than species or ecosystems, and so post-processing of existing land cover data is often required. This study compares over Siberia, multiple land cover data sets against one another and with auxiliary data to identify key uncertainties that contribute to variability in PFT classifications that would introduce errors in Earth system modeling. Land cover classification systems from GLC 2000, GlobCover 2005 and 2009, and MODIS collections 5 and 5.1 are first aggregated to a common legend, and then compared to high-resolution land cover classification systems, vegetation continuous fields (MODIS VCFs) and satellite-derived tree heights (to discriminate against sparse, shrub, and forest vegetation). The GlobCover data set, with a lower threshold for tree cover and taller tree heights and a better spatial resolution, tends to have better distributions of tree cover compared to high-resolution data. It has therefore been chosen to build new PFT maps for the ORCHIDEE land surface model at 1 km scale. Compared to the original PFT data set, the new PFT maps based on GlobCover 2005 and an updated cross-walking approach mainly differ in the characterization of forests and degree of tree cover. The partition of grasslands and bare soils now appears more realistic compared with ground truth data. This new vegetation map provides a framework for further development of new PFTs in the ORCHIDEE model like shrubs, lichens and mosses, to represent the water and carbon cycles in northern latitudes better. Updated land cover data sets are critical for improving and maintaining the relevance of Earth system models for assessing climate and human impacts on biogeochemistry and biophysics.

Sedimentverteilung im Seegebiet bei Bokniseck, Eckernförder Bucht, westliche Ostsee (sediment distribution at Bokniseck, western Baltic Sea; map)

A detailed topographic survey was carried out in the "Hausgarten"-area of the Joint Research Programm 95 of the University of Kiel by the Deutsches Hydrographisches Institut. Based on this information a sediment distribution map was constructed. A horizontal section extending from 0 to 27 m of water depth was investigated showing the distribution of pebbles and boulders, of algal growth, and exposed areas of glacial marl; the grain size distribution was determined for the various sediment types.

Pan-European land cover/use map for 2009

GlobCorine demonstrated an automatic service that can generate in a consistent way land cover / land use maps and land change indicators, based on a CLC-compatible legend. CLC is derived from a visual identification and classification of landscape objects using high resolution images. This methodology provides high thematic accuracy but limits the update rate since it is time-consuming. Therefore, the project evaluated the use of MERIS FR time series, processed automatically to provide a more frequent update of CLC-compatible maps. GlobCorine built upon the experience and resources available through the GlobCover project, to tune the classification chain and adapt it to the EEA needs, covering the pan-European area (including the Mediterranean basin and the European Russia), although the system could be potentially extendable globally. The project delivered two CLC-compatible pan-European land cover maps in less than two years, demonstrating efficient and quick production. The first map is based on Envisat MERIS fine resolution (300m) mode data acquired between end 2004 and mid 2006, while the second used full-year 2009 data. GlobCorine is an initiative of ESA with the partnership of EEA and is implemented by Universite' catholique de Louvain - UCL.

Climate and total tree cover reconstructions based on the 43.7-kyr fossil pollen record from Khoe (NW Sakhalin Island) and modern climate variables of 236 modern surface pollen sampling sites

A late Quaternary pollen record from northern Sakhalin Island (51.34°N, 142.14°E, 15 m a.s.l.) spanning the last 43.7 ka was used to reconstruct regional climate dynamics and vegetation distribution by using the modern analogue technique (MAT). The long-term trends of the reconstructed mean annual temperature (TANN) and precipitation (PANN), and total tree cover are generally in line with key palaeoclimate records from the North Atlantic region and the Asian monsoon domain. TANN largely follows the fluctuations in solar summer insolation at 55°N. During Marine Isotope Stage (MIS) 3, TANN and PANN were on average 0.2 °C and 700 mm, respectively, thus very similar to late Holocene/modern conditions. Full glacial climate deterioration (TANN = -3.3 °C, PANN = 550 mm) was relatively weak as suggested by the MAT-inferred average climate parameters and tree cover densities. However, error ranges of the climate reconstructions during this interval are relatively large and the last glacial environments in northern Sakhalin could be much colder and drier than suggested by the weighted average values. An anti-phase relationship between mean temperature of the coldest (MTCO) and warmest (MTWA) month is documented during the last glacial period, i.e. MIS 2 and 3, suggesting more continental climate due to sea levels that were lower than present. Warmest and wettest climate conditions have prevailed since the end of the last glaciation with an optimum (TANN = 1.5 °C, PANN = 800 mm) in the middle Holocene interval (ca 8.7-5.2 cal. ka BP). This lags behind the solar insolation peak during the early Holocene. We propose that this is due to continuous Holocene sea level transgression and regional influence of the Tsushima Warm Current, which reached maximum intensity during the middle Holocene. Several short-term climate oscillations are suggested by our reconstruction results and correspond to Northern Hemisphere Heinrich and Dansgaard-Oeschger events, the Bølling-Allerød and the Younger Dryas. The most prominent fluctuation is registered during Heinrich 4 event, which is marked by noticeably colder and drier conditions and the spread of herbaceous taxa.

Forest disturbance map for the Carpathian ecoregion, 1984-2012

Detailed knowledge of forest cover dynamics is crucial for many applications from resource management to ecosystem service assessments. Landsat data provides the necessary spatial, temporal and spectral detail to map and analyze forest cover and forest change processes. With the opening of the Landsat archive, new opportunities arise to monitor forest dynamics on regional to continental scales. In this study we analyzed changes in forest types, forest disturbances, and forest recovery for the Carpathian ecoregion in Eastern Europe. We generated a series of image composites at five year intervals between 1985 and 2010 and utilized a hybrid analysis strategy consisting of radiometric change classification, post-classification comparison and continuous index- and segment-based post-disturbance recovery assessment. For validation of the disturbance map we used a point-based accuracy assessment, and assessed the accuracy of our forest type maps using forest inventory data and statistically sampled ground truth data for 2010. Our Carpathian-wide disturbance map achieved an overall accuracy of 86% and the forest type maps up to 73% accuracy. While our results suggested a small net forest increase in the Carpathians, almost 20% of the forests experienced stand-replacing disturbances over the past 25 years. Forest recovery seemed to only partly counterbalance the widespread natural disturbances and clear-cutting activities. Disturbances were most widespread during the late 1980s and early 1990s, but some areas also exhibited extensive forest disturbances after 2000, especially in the Polish, Czech and Romanian Carpathians. Considerable shifts in forest composition occurred in the Carpathians, with disturbances increasingly affecting coniferous forests, and a relative decrease in coniferous and mixed forests. Both aspects are likely connected to an increased vulnerability of spruce plantations to pests and pathogens in the Carpathians. Overall, our results exemplify the highly dynamic nature of forest cover during times of socio-economic and institutional change, and highlight the value of the Landsat archive for monitoring these dynamics.