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By overcoming the diffraction limit in light microscopy, super-resolution techniques, such as stimulated emission depletion (STED) microscopy, are experiencing an increasing impact on life sciences. High costs and technically demanding setups, however, may still hinder a wider distribution of this innovation in biomedical research laboratories. As far-field microscopy is the most widely employed microscopy modality in the life sciences, upgrading already existing systems seems to be an attractive option for achieving diffraction-unlimited fluorescence microscopy in a cost-effective manner. Here, we demonstrate the successful upgrade of a commercial time-resolved confocal fluorescence microscope to an easy-to-align STED microscope in the single-beam path layout, previously proposed as "easy-STED", achieving lateral resolution <lambda/10 corresponding to a five-fold improvement over a confocal modality. For this purpose, both the excitation and depletion laser beams pass through a commercially available segmented phase plate that creates the STED-doughnut light distribution in the focal plane, while leaving the excitation beam unaltered when implemented into the joint beam path. Diffraction-unlimited imaging of 20 nm-sized fluorescent beads as reference were achieved with the wavelength combination of 635 nm excitation and 766 nm depletion. To evaluate the STED performance in biological systems, we compared the popular phalloidin-coupled fluorescent dyes Atto647N and Abberior STAR635 by labeling F-actin filaments in vitro as well as through immunofluorescence recordings of microtubules in a complex epithelial tissue. Here, we applied a recently proposed deconvolution approach and showed that images obtained from time-gated pulsed STED microscopy may benefit concerning the signal-to-background ratio, from the joint deconvolution of sub-images with different spatial information which were extracted from offline time gating.
Studies investigating the effect of increasing CO2 levels on the phosphorus cycle in natural waters are lacking although phosphorus often controls phytoplankton development in many aquatic systems. The aim of our study was to analyse effects of elevated CO2 levels on phosphorus pool sizes and uptake. The phosphorus dynamic was followed in a CO2-manipulation mesocosm experiment in the Storfjarden (western Gulf of Finland, Baltic Sea) in summer 2012 and was also studied in the surrounding fjord water. In all mesocosms as well as in surface waters of Storfjarden, dissolved organic phosphorus (DOP) concentrations of 0.26aEuro-+/- aEuro-0.03 and 0.23aEuro-+/- aEuro-0.04aEuro-A mu molaEuro-L-1, respectively, formed the main fraction of the total P-pool (TP), whereas phosphate (PO4) constituted the lowest fraction with mean concentration of 0.15aEuro-A +/- aEuro-0.02 in the mesocosms and 0.17aEuro-A +/- aEuro-0.07aEuro-A mu molaEuro-L-1 in the fjord. Transformation of PO4 into DOP appeared to be the main pathway of PO4 turnover. About 82aEuro-% of PO4 was converted into DOP whereby only 18aEuro-% of PO4 was transformed into particulate phosphorus (PP). PO4 uptake rates measured in the mesocosms ranged between 0.6 and 3.9aEuro-nmolaEuro-L(-1)aEuro-h(-1). About 86aEuro-% of them was realized by the size fraction < aEuro-3aEuro-A mu m. Adenosine triphosphate (ATP) uptake revealed that additional P was supplied from organic compounds accounting for 25-27aEuro-% of P provided by PO4 only. CO2 additions did not cause significant changes in phosphorus (P) pool sizes, DOP composition, and uptake of PO4 and ATP when the whole study period was taken into account. However, significant short-term effects were observed for PO4 and PP pool sizes in CO2 treatments > aEuro-1000aEuro-A mu atm during periods when phytoplankton biomass increased. In addition, we found significant relationships (e.g., between PP and Chl a) in the untreated mesocosms which were not observed under high fCO(2) conditions. Consequently, it can be hypothesized that the relationship between PP formation and phytoplankton growth changed with CO2 elevation. It can be deduced from the results, that visible effects of CO2 on P pools are coupled to phytoplankton growth when the transformation of PO4 into POP was stimulated. The transformation of PO4 into DOP on the other hand does not seem to be affected. Additionally, there were some indications that cellular mechanisms of P regulation might be modified under CO2 elevation changing the relationship between cellular constituents.
Introduction
Genes involved in body weight regulation that were previously investigated in genome-wide association studies (GWAS) and in animal models were target-enriched followed by massive parallel next generation sequencing.
Methods
We enriched and re-sequenced continuous genomic regions comprising FTO, MC4R, TMEM18, SDCCAG8, TKNS, MSRA and TBC1D1 in a screening sample of 196 extremely obese children and adolescents with age and sex specific body mass index (BMI) >= 99th percentile and 176 lean adults (BMI <= 15th percentile). 22 variants were confirmed by Sanger sequencing. Genotyping was performed in up to 705 independent obesity trios (extremely obese child and both parents), 243 extremely obese cases and 261 lean adults.
Results and Conclusion
We detected 20 different non-synonymous variants, one frame shift and one nonsense mutation in the 7 continuous genomic regions in study groups of different weight extremes. For SNP Arg695Cys (rs58983546) in TBC1D1 we detected nominal association with obesity (p(TDT) = 0.03 in 705 trios). Eleven of the variants were rare, thus were only detected heterozygously in up to ten individual(s) of the complete screening sample of 372 individuals. Two of them (in FTO and MSRA) were found in lean individuals, nine in extremely obese. In silico analyses of the 11 variants did not reveal functional implications for the mutations. Concordant with our hypothesis we detected a rare variant that potentially leads to loss of FTO function in a lean individual. For TBC1D1, in contrary to our hypothesis, the loss of function variant (Arg443Stop) was found in an obese individual. Functional in vitro studies are warranted.
Solving problems combining task and motion planning requires searching across a symbolic search space and a geometric search space. Because of the semantic gap between symbolic and geometric representations, symbolic sequences of actions are not guaranteed to be geometrically feasible. This compels us to search in the combined search space, in which frequent backtracks between symbolic and geometric levels make the search inefficient. We address this problem by guiding symbolic search with rich information extracted from the geometric level through culprit detection mechanisms.
Siberian arctic vegetation and lake water communities, known for their temperature dependence, are expected to be particularly impacted by recent climate change and high warming rates. However, decadal information on the nature and strength of recent vegetation change and its time lag to climate signals are rare. In this study, we present a Pb-210/Cs-137 dated pollen and Pediastrum species record from a unnamed lake in the south of the Taymyr peninsula covering the period from AD 1706 to 2011. Thirty-nine palynomorphs and 10 morphotypes of Pediastrum species were studied to assess changes in vegetation and lake conditions as probable responses to climate change. We compared the pollen record with Pediastrum species, which we consider to be important proxies of climate changes. Three pollen assemblage zones characterised by Betula nana, Alnus viridis and Larix gmelinii (1706-1808); herbs such as Cyperaceae, Artemisia or Senecio (1808-1879), and higher abundance of Larix pollen (1955-2011) are visible. Also, three Pediastrum assemblage zones show changes of aquatic conditions: higher abundances of Pediastrum boryanum var. brevicorne (1706-1802); medium abundances of P. kawraiskyi and P. integrum (1802-1840 and 1920-1980), indicating cooler conditions while less eutrophic conditions are indicated by P. boryanum, and a mainly balanced composition with only small changes of cold- and warm-adapted Pediastrum species (1965-2011). In general, compositional Pediastrum species turnover is slightly higher than that indicated by pollen data (0.54 vs 0.34 SD), but both are only minor for this treeline location. In conclusion, the relevance of differentiation of Pediastrum species is promising and can give further insights into the relationship between lakes and their surrounding vegetation transferred onto climatic conditions.
This study investigates the spatial and temporal distributions of 14 key arboreal taxa and their driving forces during the last 22,000 calendar years before ad 1950 (kyr BP) using a taxonomically harmonized and temporally standardized fossil pollen dataset with a 500-year resolution from the eastern part of continental Asia. Logistic regression was used to estimate pollen abundance thresholds for vegetation occurrence (presence or dominance), based on modern pollen data and present ranges of 14 taxa in China. Our investigation reveals marked changes in spatial and temporal distributions of the major arboreal taxa. The thermophilous (Castanea, Castanopsis, Cyclobalanopsis, Fagus, Pterocarya) and eurythermal (Juglans, Quercus, Tilia, Ulmus) broadleaved tree taxa were restricted to the current tropical or subtropical areas of China during the Last Glacial Maximum (LGM) and spread northward since c. 14.5 kyr BP. Betula and conifer taxa (Abies, Picea, Pinus), in contrast, retained a wider distribution during the LGM and showed no distinct expansion direction during the Late Glacial. Since the late mid-Holocene, the abundance but not the spatial extent of most trees decreased. The changes in spatial and temporal distributions for the 14 taxa are a reflection of climate changes, in particular monsoonal moisture, and, in the late Holocene, human impact. The post-LGM expansion patterns in eastern continental China seem to be different from those reported for Europe and North America, for example, the westward spread for eurythermal broadleaved taxa.
John Birks
(2015)
We describe the career of John Birks as a pioneering scientist who has, over a career spanning five decades, transformed palaeoecology from a largely descriptive to a rigorous quantitative science relevant to contemporary questions in ecology and environmental change. We review his influence on students and colleagues not only at Cambridge and Bergen Universities, his places of primary employment, but also on individuals and research groups in Europe and North America. We also introduce the collection of papers that we have assembled in his honour. The papers are written by his former students and close colleagues and span many of the areas of palaeoecology to which John himself has made major contributions. These include the relationship between ecology and palaeoecology, late-glacial and Holocene palaeoecology, ecological succession, climate change and vegetation history, the role of palaeoecological techniques in reconstructing and understanding the impact of human activity on terrestrial and freshwater ecosystems and numerical analysis of multivariate palaeoecological data.
Efficacy, Safety & Modification of Albuminuria in Type 2 Diabetes Subjects with Renal Disease with LINAgliptin (MARLINA-T2D), a multicentre, multinational, randomized, double-blind, placebo-controlled, parallel-group, phase 3b clinical trial, aims to further define the potential renal effects of dipeptidyl peptidase-4 inhibition beyond glycaemic control. A total of 350 eligible individuals with inadequately controlled type 2 diabetes and evidence of renal disease are planned to be randomized in a 1:1 ratio to receive either linagliptin 5mg or placebo in addition to their stable glucose-lowering background therapy for 24weeks. Two predefined main endpoints will be tested in a hierarchical manner: (1) change from baseline in glycated haemoglobin and (2) time-weighted average of percentage change from baseline in urinary albumin-to-creatinine ratio. Both endpoints are sufficiently powered to test for superiority versus placebo after 24weeks with =0.05. MARLINA-T2D is the first of its class to prospectively explore both the glucose- and albuminuria-lowering potential of a dipeptidyl peptidase-4 inhibitor in patients with type 2 diabetes and evidence of renal disease.
This study examines the course and driving forces of recent vegetation change in the Mongolian steppe. A sediment core covering the last 55years from a small closed-basin lake in central Mongolia was analyzed for its multi-proxy record at annual resolution. Pollen analysis shows that highest abundances of planted Poaceae and highest vegetation diversity occurred during 1977-1992, reflecting agricultural development in the lake area. A decrease in diversity and an increase in Artemisia abundance after 1992 indicate enhanced vegetation degradation in recent times, most probably because of overgrazing and farmland abandonment. Human impact is the main factor for the vegetation degradation within the past decades as revealed by a series of redundancy analyses, while climate change and soil erosion play subordinate roles. High Pediastrum (a green algae) influx, high atomic total organic carbon/total nitrogen (TOC/TN) ratios, abundant coarse detrital grains, and the decrease of C-13(org) and N-15 since about 1977 but particularly after 1992 indicate that abundant terrestrial organic matter and nutrients were transported into the lake and caused lake eutrophication, presumably because of intensified land use. Thus, we infer that the transition to a market economy in Mongolia since the early 1990s not only caused dramatic vegetation degradation but also affected the lake ecosystem through anthropogenic changes in the catchment area.
Recent research has called into question the current practice to estimate individual usual food intake in large-scale studies. In such studies, usual food intake has been defined as diet over the past year. The aim of this review is to summarise the concepts of dietary assessment methods providing food intake data over this time period. A conceptualised framework is given to help researchers to understand the more recent developments to improve dietary assessment in large-scale prospective studies, and also to help to spot the gaps that need to be addressed in future methodological research. The conceptual framework illustrates the current options for the assessment of an individual’s food consumption over 1 year. Ideally, a person’s food intake on each day of this year should be assessed. Due to participants’ burden, and organisational and financial constraints, however, the options are limited to directly requesting the long-term average (e.g. food frequency questionnaires), or selecting a few days with detailed food consumption measurements (e.g. 24-hour dietary recalls) or using snapshot techniques (e.g. barcode scanning of purchases). It seems necessary and important to further evaluate the performance of statistical modelling of the individual usual food intake from all available sources. Future dietary assessment might profit from the growing prominence of internet and telecommunication technologies to further enhance the available data on food consumption for each study participant. Research is crucial to investigate the performance of innovative assessment tools. However, the self-reported nature of the data itself will always lead to bias.