Expression of a cDNA encoding the glucose trimming enzyme glucosidase II in CHO cells and molecular characterization of the enzyme deficiency in a mutant mouse lymphoma cell line

Glucosidase II is an ER resident glycoprotein involved in the processing of N-linked glycans and probably a component of the ER quality control of glycoproteins. For cloning of glucosidase II cDNA, degenerate oligonucleotides based on amino acid sequences derived from proteolytic fragments of purified pig liver glucosidase II were used. An unamplified cDNA library from pig liver was screened with a 760 bp glucosidase II specific cDNA fragment obtained by RT-PCR. A 3.9 kb glucosidase II cDNA with an open reading frame of about 2.9 kb was obtained. The glucosidase II sequence did not contain known ER retention signals nor hydrophobic regions which could represent a transmem-brane domain; however, it contained a single N-glycosylation site close to the amino terminus. All studied pig and rat tissues exhibited an mRNA of approximately 4.4 kb with varying tissue expression levels. The authenticity of the identified cDNA with that coding for glucosidase II was proven by overexpression in CHO cells. Mouse lymphoma PHAR 2.7 cells, deficient in glucosidase II activity, were shown to be devoid of transcript

Symbiotische Stickstofffixierung in biologisch und konventionell bewirtschafteten Wiesen

Because of lower nitrogen (N) input it is assumed that organically cultivated legumes fix more N2 than legumes grown under conventional cropping. Using the natural 15N abundance method, we assessed symbiotic N2 fixation by white and red clover. The clover was growing in a grass-clover meadow installed as part of the crop rotation of a long term (30 years) field experiment. Dry matter yields were similar for organic and conventional meadows on plots fertilized at levels typical for the respective system, but organic meadows tended to higher clover yields. The proportion of N derived from atmosphere (Ndfa) in both clover species was on average 84%. It was not significantly affected by the cropping system. Because of more legumes, amounts of fixed N2 (Nfix) were higher in organic than conventional meadows. Under low fertilization intensity, low available potassium and phosphorus contents limited Nfix through lower legume dry matter production while Ndfa remained high

Increased abundance of MTD1 and MTD2 mRNAs in nodules of decapitated Medicago truncatula.

To gain insight into the molecular processes occurring in root nodule metabolism after stress, we used a mRNA differential display (DDRT-PCR) approach to identify cDNAs corresponding to genes whose expression is enhanced in nodules of decapitated Medicago truncatula plants. Two full-length cDNAs of plant origin were isolated (MTD1 and MTD2). Sequence analysis revealed that MTD1 is identical to an EST clone (accession number AW559774) expressed in roots of M. truncatula upon infection with Phytophthora medicaginis, while MTD2 is highly homologous to an Arabidopsis thaliana gene (accession number AL133292) coding for a RNA binding-like protein. The two mRNAs started to accumulate in root nodules at 4 h after plant decapitation and reached even higher transcript levels at 24 h from the imposition of the treatment. MTD1 and MTD2 mRNAs were mainly induced in nodules, with very little induction in roots. The abundance of the two transcripts did not change in response to other perturbations known to decrease nitrogenase activity, such as nitrate and Ar/O2 treatments. Our results suggest that MTD1 and MTD2 represent transcripts that accumulate locally in nodules and may be involved in changes in nodule metabolism in response to decapitation

ECLAIRE: Effects of Climate Change on Air Pollution Impacts and Response Strategies for European Ecosystems. Project final report

The central goal of ECLAIRE is to assess how climate change will alter the extent to which air pollutants threaten terrestrial ecosystems. Particular attention has been given to nitrogen compounds, especially nitrogen oxides (NOx) and ammonia (NH3), as well as Biogenic Volatile Organic Compounds (BVOCs) in relation to tropospheric ozone (O3) formation, including their interactions with aerosol components. ECLAIRE has combined a broad program of field and laboratory experimentation and modelling of pollution fluxes and ecosystem impacts, advancing both mechanistic understanding and providing support to European policy makers. The central finding of ECLAIRE is that future climate change is expected to worsen the threat of air pollutants on Europe’s ecosystems. Firstly, climate warming is expected to increase the emissions of many trace gases, such as agricultural NH3, the soil component of NOx emissions and key BVOCs. Experimental data and numerical models show how these effects will tend to increase atmospheric N deposition in future. By contrast, the net effect on tropospheric O3 is less clear. This is because parallel increases in atmospheric CO2 concentrations will offset the temperature-driven increase for some BVOCs, such as isoprene. By contrast, there is currently insufficient evidence to be confident that CO2 will offset anticipated climate increases in monoterpene emissions. Secondly, climate warming is found to be likely to increase the vulnerability of ecosystems towards air pollutant exposure or atmospheric deposition. Such effects may occur as a consequence of combined perturbation, as well as through specific interactions, such as between drought, O3, N and aerosol exposure. These combined effects of climate change are expected to offset part of the benefit of current emissions control policies. Unless decisive mitigation actions are taken, it is anticipated that ongoing climate warming will increase agricultural and other biogenic emissions, posing a challenge for national emissions ceilings and air quality objectives related to nitrogen and ozone pollution. The O3 effects will be further worsened if progress is not made to curb increases in methane (CH4) emissions in the northern hemisphere. Other key findings of ECLAIRE are that: 1) N deposition and O3 have adverse synergistic effects. Exposure to ambient O3 concentrations was shown to reduce the Nitrogen Use Efficiency of plants, both decreasing agricultural production and posing an increased risk of other forms of nitrogen pollution, such as nitrate leaching (NO3-) and the greenhouse gas nitrous oxide (N2O); 2) within-canopy dynamics for volatile aerosol can increase dry deposition and shorten atmospheric lifetimes; 3) ambient aerosol levels reduce the ability of plants to conserve water under drought conditions; 4) low-resolution mapping studies tend to underestimate the extent of local critical loads exceedance; 5) new dose-response functions can be used to improve the assessment of costs, including estimation of the value of damage due to air pollution effects on ecosystems, 6) scenarios can be constructed that combine technical mitigation measures with dietary change options (reducing livestock products in food down to recommended levels for health criteria), with the balance between the two strategies being a matter for future societal discussion. ECLAIRE has supported the revision process for the National Emissions Ceilings Directive and will continue to deliver scientific underpinning into the future for the UNECE Convention on Long-range Transboundary Air Pollution

ECLAIRE third periodic report

The ÉCLAIRE project (Effects of Climate Change on Air Pollution Impacts and Response Strategies for European Ecosystems) is a four year (2011-2015) project funded by the EU's Seventh Framework Programme for Research and Technological Development (FP7)

Expression of a cDNA encoding the glucose trimming enzyme glucosidase II in CHO cells and molecular characterization of the enzyme deficiency in a mutant mouse lymphoma cell line

Glucosidase II is an ER resident glycoprotein involved in the processing of N-linked glycans and probably a component of the ER quality control of glycoproteins. For cloning of glucosidase II cDNA, degenerate oligonucleotides based on amino acid sequences derived from proteolytic fragments of purified pig liver glucosidase II were used. An unamplified cDNA library from pig liver was screened with a 760 bp glucosidase II specific cDNA fragment obtained by RT-PCR. A 3.9 kb glucosidase II cDNA with an open reading frame of about 2.9 kb was obtained. The glucosidase II sequence did not contain known ER retention signals nor hydrophobic regions which could represent a transmem-brane domain; however, it contained a single N-glycosylation site close to the amino terminus. All studied pig and rat tissues exhibited an mRNA of approximately 4.4 kb with varying tissue expression levels. The authenticity of the identified cDNA with that coding for glucosidase II was proven by overexpression in CHO cells. Mouse lymphoma PHAR 2.7 cells, deficient in glucosidase II activity, were shown to be devoid of transcript

Gadolinium doped ceria interlayers for Solid Oxide Fuel Cells cathodes: Enhanced reactivity with sintering aids (Li, Cu, Zn), and improved densification by infiltration

This paper reports the study of the densification of 20% Gd doped ceria (Ce0.8Gd0.2O1.9 (GDC)) interlayers in SOFC cathodes through two different routes: the well-known addition of sintering elements, and an innovative densification process by infiltration. First, Li, Cu, and Zn nitrates were added to GDC powders. The effect of these additives on the densification was studied by dilatometry on pellets, and show a large decrease of the sintering temperature from 1330 °C (pure GDC), down to 1080 °C, 950 °C, and 930 °C for Zn, Cu, and Li addition, respectively. However, this promising result does not apply to screen-printed layers, which are more porous than pellets and in which the shrinkage is constrained by the substrate. The second approach consists in preparing a pre-sintered GDC layer, which is subsequently infiltrated with Ce and Gd nitrates and sintered at 1250 °C to increase its density. Such an approach results in highly dense GDC interlayers. Using La0.6Sr0.4Co0.2Fe0.8O3-δ (LSCF) as electrode, the influence of the interlayers on the cathode performance was studied. The addition of sintering aids dramatically increases the cell resistances, most likely because the additives increase the reactivity between GDC and either Yttria Stabilized Zirconia (YSZ) or LSCF, thus losing the expected benefit related to the decrease of sintering temperatures. The interlayers prepared by infiltration do not induce additional resistances in the cell, which results in power densities of single cells 40–50% higher than those of cells prepared with commercial GDC interlayers, making this approach a valuable alternative to sintering aids