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This paper analyses what post 2020 targets may be necessary for the European CO2 legislation for passenger cars in order to reach the overall sectoral goal of 60% reduction of transport's greenhouse gas emissions by 2050 relative to 1990, as defined in the European Commission's White Paper. The required target levels are found to depend strongly on the contribution that passenger cars need to make to reaching the overall target, on the assumed growth of passenger car mobility, and on the extent to which biofuels could be available for fuelling passenger cars. To what extent electric vehicles and other low emission vehicles such as plug-in hybrids and fuel cell electric vehicles are needed to meet the post-2020 targets depends on the target level and on the minimum CO2 emissions that can be reached in conventional vehicles. Almost all assessed scenarios, require significant amounts of low emission vehicles to be sold from 2030 or 2035 onwards. Electric and plug-in vehicles are in the early stage of market introduction now, but developments are fragile could end up in a 'valley of death'. Defining a sufficiently low passenger car CO2 target for 2025 may be an effective instrument to motivate manufacturers to continue their efforts in the marketing and further development of these vehicles. This is important for maintaining the momentum of the transition towards large-scale application of low CO2 emitting vehicles.

The causes of renewed growth in the atmospheric CH4 burden since 2007 are still poorly understood and subject of intensive scientific discussion. We present a reanalysis of global CH4 emissions during the 2000s, based on the TM5-4DVAR inverse modeling system. The model is optimized using high-accuracy surface observations from NOAA ESRL's global air sampling network for 2000-2010 combined with retrievals of column-averaged CH4 mole fractions from SCIAMACHY onboard ENVISAT (starting 2003).Using climatological OH fields, derived global total emissions for 2007-2010 are 16-20 Tg CH 4/yr higher compared to 2003-2005. Most of the inferred emission increase was located in the tropics (9-14 Tg CH4/yr) and mid-latitudes of the northern hemisphere (6-8 Tg CH4/yr), while no significant trend was derived for Arctic latitudes. The atmospheric increase can be attributed mainly to increased anthropogenic emissions, but the derived trend is significantly smaller than estimated in the EDGARv4.2 emission inventory. Superimposed on the increasing trend in anthropogenic CH4 emissions are significant inter-annual variations (IAV) of emissions from wetlands (up to ±10 Tg CH4/yr), and biomass burning (up to ±7 Tg CH4/yr). Sensitivity experiments, which investigated the impact of the SCIAMACHY observations (versus inversions using only surface observations), of the OH fields used, and of a priori emission inventories, resulted in differences in the detailed latitudinal attribution of CH4 emissions, but the IAV and trends aggregated over larger latitude bands were reasonably robust. All sensitivity experiments show similar performance against independent shipboard and airborne observations used for validation, except over Amazonia where satellite retrievals improved agreement with observations in the free troposphere. Key Points A reanalysis of global CH4 emissions during the 2000s is presented derived global total emissions 2007-2010 16-20 Tg CH4/yr higher than 2003-2005 increase mainly in the tropics and NH mid-latitudes ©2013. American Geophysical Union. All Rights Reserved.

The use of recombinant human proteins for the treatment of several diseases has increased considerably during the last decades. A major safety and efficacy issue of biopharmaceuticals is their potential immunogenicity. To prevent immunogenicity, biotechnology-derived proteins are engineered to be as human-like as possible. Immunogenicity is mainly determined in non-human primates (NHP), as they are considered to be the best predictive animal species for human safety, based on their close relatedness to man. As minipigs are increasingly used in the safety evaluation of (bio)pharmaceuticals, the predictive value of the minipig in immunogenicity testing was evaluated in this study, using anakinra as a model compound. Animals were treated subcutaneously with either placebo, low-(0.5mg/kg), or high-dose (5mg/kg) anakinra daily on 29 consecutive days. After the first and last dose, the pharmacokinetic (PK) profile of anakinra was evaluated. Antibodies directed to anakinra were measured on several time points during the treatment period. Furthermore, hematology, clinical chemistry, body weight, clinical signs, and histopathology of several organs were evaluated. No signs of toxicity were observed upon treatment with anakinra. PK parameters were comparable with those found in human and NHP studies performed with anakinra. All animals developed anti-anakinra antibodies. The results obtained in minipigs were comparable to those observed in monkeys. For anakinra, the predictive value of the minipig for immunogenicity testing was found to be comparable to that seen in NHP. However, more studies evaluating additional biopharmaceutical products are needed to support the use of the minipig as an alternative model for (immuno)toxicity testing, including immunogenicity. © 2013 Informa Healthcare USA, Inc.

A validated in vitro model of the large intestine (TIM-2), set up with human or pig faeces, was used to evaluate the impact of potentially probiotic Lactobacillus amylovorus DSM 16698, administered alone (i), in the presence of prebiotic galactooligosaccharides (GOS) (ii), and co-administered with probiotic Bifidobacterium animalis ssp. lactis Bb-12 (Bb-12) (iii) on GOS degradation, microbial growth (L. amylovorus, lactobacilli, bifidobacteria and total bacteria) and metabolite production. High performance anion exchange chromatography revealed that GOS degradation was more pronounced in TIM-2 inoculated with pig faeces than with human faeces. Denaturing gradient gel electrophoresis profiling of PCR-amplified 16S rRNA genes detected a more complex Lactobacillus spp. community in pig faecal material than in human faecal inoculum. According to 16S rRNA gene-targeted qPCR, GOS stimulated the growth of lactobacilli and bifidobacteria in faecal material from both materials. The cumulative production of short chain fatty acids and ammonia was higher (P < 0.05) for pig than for human faeces. However, lactate accumulation was higher (P < 0.05) in the human model and increased after co-administration with GOS and Bb-12. This study reinforced the notion that differences in microbiota composition between target host organisms need to be considered when animal data are extrapolated to human, as is often done with pre- and probiotic intervention studies. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

TNO’s current research programs aim to add flexibility to current food manufacturing practices, by introducing new sustainable processes and ingredient alternatives for the food industry.

This study is of our particular interest as the quality of our chemical transport model Lotos-Euros can be improved by our understanding of the aerosol-light interaction. In this study we derive aerosol water and chemical composition by a modeling approach that combines in situ measured and remotely sensed aerosol properties with radiosonde measurements of relative humidity. The model simulates water uptake by aerosols based on the chemical composition and size distribution.

Data are essential for any empirical modelling effort. In freight transport, obtaining good data for modelling can be a challenge because of the difficulty to observe freight processes directly, the often proprietary nature of logistics data, and a relatively poor data acquisition infrastructure. In this chapter we review the different data sources for freight modelling, including data for trade, logistics and transport by different modes of transport. We discuss which sources can be used for which models. In every practical modelling situation, data limitations will remain. We discuss two main strategies to overcome them: adapting the model to a simplified form and estimating the missing data from the available information. The chapter concludes with listing some challenges for research into data acquisition for the future. © 2014 Elsevier Inc. All rights reserved.

Lignocellulosic biomass is the future feedstock for the production of biofuel and bio-based chemicals. The pretreatment-hydrolysis product of biomass, so-called hydrolysate, contains not only fermentable sugars, but also compounds that inhibit its fermentability by microbes. To reduce the toxicity of hydrolysates as fermentation media, knowledge of the identity of inhibitors and their dynamics in hydrolysates need to be obtained. In the past decade, various studies have applied targeted metabolomics approaches to examine the composition of biomass hydrolysates. In these studies, analytical methods like HPLC, RP-HPLC, CE, GC-MS and LC-MS/MS were used to detect and quantify small carboxylic acids, furans and phenols. Through applying targeted metabolomics approaches, inhibitors were identified in hydrolysates and their dynamics in fermentation processes were monitored. However, to reveal the overall composition of different hydrolysates and to investigate its influence on hydrolysate fermentation performance, a non-targeted metabolomics study needs to be conducted. In this review, a non-targeted and generic metabolomics approach is introduced to explore inhibitor identification in biomass hydrolysates, and other similar metabolomics questions.

The atmospheric concentration of elemental carbon (EC) in Europe during the six-year period 2005-2010 has been simulated with the EMEP MSC-W model. The model bias compared to EC measurements was less than 20% for most of the examined sites. The model results suggest that fossil fuel combustion is the dominant source of EC in most of Europe but that there are important contributions also from residential wood burning during the cold seasons and, during certain episodes, also from open biomass burning (wildfires and agricultural fires). The modelled contributions from open biomass fires to ground level concentrations of EC were small at the sites included in the present study, EC measurements and modelled EC were also compared to optical measurements of black carbon (BC). The relationships between EC and BC (as given by mass absorption cross section, MAC, values) differed widely between the sites, and the correlation between observed EC and BC is sometimes poor, making it difficult to compare results using the two techniques and limiting the comparability of BC measurements to model EC results. A new bottom-up emission inventory for carbonaceous aerosol from residential wood combustion has been applied. For some countries the new inventory has substantially different EC emissions compared to earlier estimates. For northern Europe the most significant changes are much lower emissions in Norway and higher emissions in neighbouring Sweden and Finland. For Norway and Sweden, comparisons to source-apportionment data from winter campaigns indicate that the new inventory may improve model-calculated EC from wood burning. Finally, three different model setups were tested with variable atmospheric lifetimes of EC in order to evaluate the model sensitivity to the assumptions regarding hygroscopicity and atmospheric ageing of EC. The standard ageing scheme leads to a rapid transformation of the emitted hydrophobic EC to hygroscopic particles, and generates similar results when assuming that all EC is aged at the point of emission. Assuming hydrophobic emissions and no ageing leads to higher EC concentrations. For the more remote sites, the observed EC concentration was in between the modelled EC using standard ageing and the scenario treating EC as hydrophobic. This could indicate too-rapid EC ageing in the model in relatively clean parts of the atmosphere. © Author(s) 2013. CC Attribution 3.0 License.

Air–liquid interface (ALI) exposures enable in vitro testing ofmixtures of gases and particles such as diesel exhaust (DE). The main objective of this study was to investigate the feasibility of exposing human lung epithelial cells at the ALI to complete DE generated by a heavy-duty truck in the state-of-the-art TNO powertrain test center. A549 cells were exposed at the air–liquid interface to DE generated by a heavy-duty Euro III truck for 1.5 h. The truck was tested at a speed of 70 km/h-1 to stimulate free-flowing traffic on a motorway. Twenty-four hours after exposure, cells were analyzed for markers of oxidative stress (GSH and HO-1), cytotoxicity (LDH) and Alamar Blue assay) and inflammation (IL-8). DE exposure resulted in an increased oxidative stress response (significantly increased HO-1 levels and significantly reduced GSH/GSSH ratio), and a decreased cell viability (significantly decreased Alamar Blue levels and slightly increased LDH levels). However, the pro-inflammatory response seemed to decrease (decrease in IL-8). The results presented here demonstrate that we are able to successfully expose A549 cells at ALI to complete DE generated by a heavy-duty truck in TNO’s powertrain test center and show oxidative stress and cytotoxicity responses due to DE exposure

Background:Chronic metabolic overload results in lipid accumulation and subsequent inflammation in white adipose tissue (WAT), often accompanied by non-alcoholic fatty liver disease (NAFLD). In response to metabolic overload, the expression of genes involved in lipid metabolism and inflammatory processes is adapted. However, it still remains unknown how these adaptations in gene expression in expanding WAT and liver are orchestrated and whether they are interrelated.Methodology/Principal Findings:ApoE*3Leiden mice were fed HFD or chow for different periods up to 12 weeks. Gene expression in WAT and liver over time was evaluated by micro-array analysis. WAT hypertrophy and inflammation were analyzed histologically. Bayesian hierarchical cluster analysis of dynamic WAT gene expression identified groups of genes ('clusters') with comparable expression patterns over time. HFD evoked an immediate response of five clusters of 'lipid metabolism' genes in WAT, which did not further change thereafter. At a later time point (>6 weeks), inflammatory clusters were induced. Promoter analysis of clustered genes resulted in specific key regulators which may orchestrate the metabolic and inflammatory responses in WAT. Some master regulators played a dual role in control of metabolism and inflammation. When WAT inflammation developed (>6 weeks), genes of lipid metabolism and inflammation were also affected in corresponding livers. These hepatic gene expression changes and the underlying transcriptional responses in particular, were remarkably similar to those detected in WAT.Conclusion:In WAT, metabolic overload induced an immediate, stable response on clusters of lipid metabolism genes and induced inflammatory genes later in time. Both processes may be controlled and interlinked by specific transcriptional regulators. When WAT inflammation began, the hepatic response to HFD resembled that in WAT. In all, WAT and liver respond to metabolic overload by adaptations in expression of gene clusters that control lipid metabolism and inflammatory processes in an orchestrated and interrelated manner. © 2013 Liang et al.

ULTimateCO2 will assess the long-term CO2 storage behaviour in terms of efficiency and security. The project is dedicated to studying the main physical processes needed to develop a better, quantitative understanding of the longterm geological storage of CO2, namely: (i) reservoir trapping, (ii) sealing integrity of caprock, and (iii) well leakage. Close collaboration with the NER300 candidates and EEPR demonstration sites will underpin all investigation with relevant supply of data industrial context. ULTimateCO2 will define a set of recommendations that will enable both regulators and operators to demonstrate that site specific long-term site performance will lead to permanent and safe CO2 containment.

Purpose: Osteoarthritis (OA) is associated with obesity in which altered fatty acid levels have been observed. We investigated whether the most common fatty acids in synovial fluid influence cartilage deterioration in OA. Design: Cartilage was obtained from OA patients undergoing total knee arthroplasty. Chondrocytes or cartilage explants were cultured with linoleic (n-6 polyunsaturated), oleic (monounsaturated), or palmitic (saturated) acid. After preculture, media were renewed and inflammation was simulated in half of the samples by addition of 10 ng/mL tumor necrosis factor-α (TNFα) with or without the fatty acids. Effects on lipid uptake (Oil-Red-O), cell toxicity (lactate dehydrogenase), prostaglandin-E2 (PGE2) release and gene expression for prostaglandin-endoperoxide synthase-2 (PTGS2), matrix metalloproteinase-1 (MMP1), and MMP13, and a disintegrin and metalloproteinase with thrombospondin motifs 4 were determined on chondrocytes in monolayer. Effects on glycosaminoglycan (GAG) release were evaluated on cartilage explants. Results: None of the fatty acids were cytotoxic and all were taken up by the cells, resulting in a higher amount of intracellular lipid in chondrocytes. Linoleic acid increased PGE2 production in the presence of TNFα. Oleic acid and palmitic acid inhibited MMP1 gene expression in chondrocytes stimulated with TNFα. In cartilage explants, GAG release was also inhibited by oleic acid and palmitic acid, and oleic acid decreased PTGS2 gene expression in stimulated chondrocytes. Conclusions: Linoleic acid has a pro-inflammatory effect on cartilage whereas oleic acid and palmitic acid seem to inhibit cartilage destruction. These results indicate that altered fatty acid levels may influence loss of cartilage structure in OA. © The Author(s) 2013.Chemicals/CAS: collagenase 3, 175449-82-8; glyceraldehyde 3 phosphate dehydrogenase, 9001-50-7; interstitial collagenase, 9001-12-1; lactate dehydrogenase, 9001-60-9; linoleic acid, 1509-85-9, 2197-37-7, 60-33-3, 822-17-3; oleic acid, 112-80-1, 115-06-0; palmitic acid, 57-10-3; prostaglandin E2, 363-24-6; prostaglandin synthase, 39391-18-9, 59763-19-8, 9055-65-6

Analysis of siliceous microfossils of a 79 cm long peat sediment core from Highlands Hammock State Park, Florida, revealed distinct changes in the local hydrology during the past 2,500 years. The coring site is a seasonally inundated forest where water availability is directly influenced by precipitation. Diatoms, chrysophyte statospores, sponge remains and phytoliths were counted in 25 samples throughout the core. Based on the relative abundance of diatom species, the record was subdivided into four diatom assemblage zones, which mainly reflect the hydrological state of the study site. An age-depth relationship based on radiocarbon measurements of eight samples reveals a basal age of the core of approximately 2,500 cal. yrs. BP. Two significant changes of diatom assemblage composition were found that could be linked to both, natural and anthropogenic influences. At 700 cal. yrs. BP, the diatom record documents a shift from tychoplanktonic Aulacoseira species to epiphytic Eunotia species, indicating a shortening of the hydroperiod, i. e. the time period during which a wetland is covered by water. This transition was interpreted as being triggered by natural climate change. In the middle of the twentieth century a second major turnover took place, at that time however, as a result of human impact on the park hydrology through the construction of dams and canals close to the study site. © 2011 The Author(s).

This work analyzes the synergy between two complementary unit operations - adsorbent dehumidification and drying - and presents a mixed integer nonlinear programming approach to optimize energy performance in a two-stage system. Combined with active constraint analysis, the adsorbent properties that promote energy performance are derived. Microporous adsorbents with higher sorption capacities at low vapor pressures and requiring higher regeneration temperatures are preferred for ambient air dehumidification in the first stage. For exhaust air dehumidification, mesoporous adsorbents with lower regeneration temperatures are preferred such that the exhaust air from the first regeneration stage can sufficiently regenerate them. For drying below 50 C, energy consumption reductions of about 70% are achieved compared to conventional dryers without adsorbent dehumidification depending on adsorbent properties. The results demonstrate the usefulness of superstructure optimization in matching the drying process with the capabilities of the adsorbents to enhance process synergy for improved energy efficiency. © 2013 American Chemical Society.

The origin of N-acyl ethanolamides (NAEs) in plasma is not well understood, and it is possible that NAEs are present in plasma in esterified form. To test this hypothesis, a new and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of arachidonoyl ethanolamide, 2-arachidonoyl glycerol, docosahexaenoyl ethanolamide, dihomo-γ-linolenoyl ethanolamide, oleoyl ethanolamide, palmitoyl ethanolamide, and stearoyl ethanolamide in 100 μl of human plasma using a simple acetonitrile extraction step. Using this method, we determined (i) free and esterified NAE levels in human plasma, (ii) free and esterified NAE levels in plasma of mice fed with diets with different amounts of n-3 fatty acids, and (iii) esterified NAE levels in blood cells. Murine and human plasma extracts contained 20- to 60-fold higher levels of esterified NAEs than free NAEs. Moreover, the effect of dietary n-3 fatty acids on murine free plasma NAE profiles was similar for esterified NAEs. Finally, esterified NAEs were also present in murine blood cells, and their pattern followed the same diet effect as observed for free and esterified NAEs in plasma. Together, these data point to the presence of previously ignored pools of esterified NAEs in plasma and blood cells that correlated well with free NAE levels in plasma. © 2012 Elsevier Inc. All rights reserved.

Wie zich een mening wil vormen over cameratoezicht is meestal afhankelijk van wat er in de media over wordt geschreven, waaronder in de dagbladen. Uit een media-analyse blijkt dat dagbladartikelen hoofdzakelijk positief van aard zijn omdat ze vaak schrijven over de succesvolle opsporing van dader(s) en dat cameratoezicht helpt tegen onveiligheidsgevoelens. De wetenschappelijke literatuur is hierover veel minder eensluidend. Deze media-analyse is gebaseerd op een database met 24.000 krantenartikelen over cameratoezicht uit de landelijke en regionale dagbladen. Deze artikelen zijn verzameld in het kader van een afstudeeronderzoek aan de Technische Universiteit Delft.

Background: Children with cow's milk allergy (CMA) need a cow's milk protein (CMP) free diet to prevent allergic reactions. For this, reliable allergy-information on the label of food products is essential to avoid products containing the allergen. On the other hand, both overzealous labeling and misdiagnosis that result in unnecessary elimination diets, can lead to potentially hazardous health situations. Our objective was to evaluate if excluding CMA by double-blind placebo-controlled food challenge (DBPCFC) prevents unnecessary elimination diets in the long term. Secondly, to determine the minimum eliciting dose (MED) for an acute allergic reaction to CMP in DBPCFC positive children. Methods. All children with suspected CMA under our care (Oct'05 - Jun'09) were prospectively enrolled in a DBPCFC. Placebo and verum feedings were administered on two randomly assigned separate days. The MED was determined by noting the 'lowest observed adverse effect level' (LOAEL) in DBPCFC-positive children. Based on the outcomes of the DBPCFC a dietary advice was given. Parents were contacted by phone several months later about the diet of their child. Results: 116 children were available for analysis. In 76 children CMA was rejected. In 60 of them CMP was successfully reintroduced, in 2 the parents refused introduction, in another 3 the parents stopped reintroduction. In 9 children CMA symptoms reappeared. In 40 children CMA was confirmed. Infants aged ≤ 12 months in our study group have a higher cumulative distribution of MED than older children. Conclusions: Excluding CMA by DBPCFC successfully stopped unnecessary elimination diets in the long term in most children. The MEDs form potential useful information for offering dietary advice to patients and their caretakers. © 2013 Dambacher et al; licensee BioMed Central Ltd.

Aanvullend onderzoek laat zien dat er binnen steden grote ruimtelijke concentratieverschillen zijn voor roet en zware metalen

Background Children’s current physical activity levels are disturbingly low when compared to recommended levels. This may be changed by intervening in the school environment. However, at present, it is unclear to what extent schoolyard physical activity contributes towards reaching the daily physical activity guideline. The aim of this study was to examine how long and at what intensity children are physically active at the schoolyard during different time segments of the day. Moreover, the contribution of schoolyard physical activity towards achieving the recommended guideline for daily physical activity was investigated. Methods Children (n=76) between the age of 6–11 years were recruited in six different schools in five cities (>70.000 residents) in the Netherlands. During the weekdays of a regular school week, childrens’ physical activity and location were measured with ActiGraph accelerometers and Travelrecorder GPS receivers. Data was collected from December 2008 to April 2009. From the data, the amount of moderate to vigorous physical activity (MVPA) on and outside the schoolyard was established. Moreover, the percentage of MVPA on the schoolyard was compared between the following segments of the day: pre-school, school, school recess, lunch break and post-school. Differences between boys and girls were compared using linear and logistic mixed-effects models. Results On average, children spent 40.1 minutes/day on the schoolyard. During this time, boys were more active on the schoolyard, with 27.3% of their time spent as MVPA compared to 16.7% among girls (OR=2.11 [95% CI 1.54 - 2.90]). The children were most active on the schoolyard during school recess, during which boys recorded 39.5% and girls recorded 23.4% of the time as MVPA (OR=2.55 [95% CI: 1.69 - 3.85]). Although children were only present at the schoolyard for 6.1% of the total reported time, this time contributed towards 17.5% and 16.8% of boys’ and girls’ minutes of MVPA. Conclusions On the schoolyard, children’s physical activity levels are higher than on average over the whole day. Physical activity levels are particularly high during school recess. The school environment seems to be an important setting for improving children’s physical activity levels. Further research on the facilitators of these high activity levels may provide targets for further promotion of physical activity among children.

In river basins, soils, groundwater, riparian zones and floodplains, streams, rivers, lakes and reservoirs act as successive filters in which the hydrology, ecology and biogeochemical processing are strongly coupled and together act to retain a significant fraction of the nutrients transported. This paper compares existing river ecology concepts with current approaches to describe river biogeochemistry, and assesses the value of these concepts and approaches for understanding the impacts of interacting global change disturbances on river biogeochemistry. Through merging perspectives, concepts, and modeling techniques, we propose integrated model approaches that encompass both aquatic and terrestrial components in heterogeneous landscapes. In this model framework, existing ecological and biogeochemical concepts are extended with a balanced approach for assessing nutrient and sediment delivery, on the one hand, and nutrient in-stream retention on the other hand. © Author(s) 2013. CC Attribution 3.0 License.

Aspergillus niger has an extraordinary potential to produce organic acids as proven by its application in industrial citric acid production. Previously, it was shown that expression of the cis-aconitate decarboxylase gene (cadA) from Aspergillus terreus converted A. niger into an itaconic acid producer (Li et al., Fungal Genet Bio 48: 602-611, 2011). After some initial steps in production optimization in the previous research (Li et al., BMC biotechnol 12: 57, 2012), this research aims at modifying host strains and fermentation conditions to further improve itaconic acid production. Expression of two previously identified A. terreus genes encoding putative organic acid transporters (mttA, mfsA) increased itaconic acid production in an A. niger cis-aconitate decarboxylase expressing strain. Surprisingly, the production did not increase further when both transporters were expressed together. Meanwhile, oxalic acid was accumulated as a by-product in the culture of mfsA transformants. In order to further increase itaconic acid production and eliminate by-product formation, the non-acidifying strain D15#26 and the oxaloacetate acetylhydrolase (oahA) deletion strain AB 1.13 ΔoahA #76 have been analyzed for itaconic acid production. Whereas cadA expression in AB 1.13 ΔoahA #76 resulted in higher itaconic acid production than strain CAD 10.1, this was not the case in strain D15#26. As expected, oxalic acid production was eliminated in both strains. In a further attempt to increase itaconic acid levels, an improved basal citric acid-producing strain, N201, was used for cadA expression. A selected transformant (N201CAD) produced more itaconic acid than strain CAD 10.1, derived from A. niger strain AB1.13. Subsequently, we have focused on the influence of dissolved oxygen (D.O.) on itaconic acid production. Interestingly, reduced D.O. levels (10-25 %) increased itaconic acid production using strain N201 CAD. Similar results were obtained in strain AB 1.13 CAD + HBD2.5 (HBD 2.5) which overexpressed a fungal hemoglobin domain. Our results showed that overexpression of the hemoglobin domain increased itaconic acid production in A. niger at lower D.O. levels. Evidently, the lower levels of D.O. have a positive influence on itaconic acid production in A. niger strains.Chemicals/CAS: itaconic acid, 97-65-4; oxygen, 7782-44-7; Oxygen, 7782-44-7; Succinates; itaconic acid, 97-65-4

Metabolites have played an essential role in our understanding of life, health, and disease for thousands of years. This domain became much more important after the concept of metabolism was discovered. In the 1950s, mass spectrometry was coupled to chromatography and made the technique more application-oriented and allowed the development of new profiling technologies. Since 1980, TNO has performed system-based metabolic profiling of body fluids, and combined with pattern recognition has led to many discoveries and contributed to the field known as metabolomics and systems biology. This review describes the development of related concepts and applications at TNO in the biomedical, pharmaceutical, nutritional, and microbiological fields, and provides an outlook for the future. © 2013 Wiley Periodicals, Inc. Rapid Commun. Mass Spectrom. 32: 399-415, 2013. © 2013 Wiley Periodicals, Inc.

Conglutins, the major peanut allergens, Ara h 2 and Ara h 6, are highly structured proteins stabilized by multiple disulfide bridges and are stable towards heat-denaturation and digestion. We sought a way to reduce their potent allergenicity in view of the development of immunotherapy for peanut allergy. Isoforms of conglutin were purified, reduced with dithiothreitol and subsequently alkylated with iodoacetamide. The effect of this modification was assessed on protein folding and IgE-binding. We found that all disulfide bridges were reduced and alkylated. As a result, the secondary structure lost α-helix and gained some β-structure content, and the tertiary structure stability was reduced. On a functional level, the modification led to a strongly decreased IgE-binding. Using conditions for limited reduction and alkylation, partially reduced and alkylated proteins were found with rearranged disulfide bridges and, in some cases, intermolecular cross-links were found. Peptide mass finger printing was applied to control progress of the modification reaction and to map novel disulfide bonds. There was no preference for the order in which disulfides were reduced, and disulfide rearrangement occurred in a non-specific way. Only minor differences in kinetics of reduction and alkylation were found between the different conglutin isoforms. We conclude that the peanut conglutins Ara h 2 and Ara h 6 can be chemically modified by reduction and alkylation, such that they substantially unfold and that their allergenic potency decreases. © 2013 Elsevier B.V.

The term “natural” in food labelling is increasingly used by producers to indicate that their products are “natural”. The use of this term is not well regulated in many countries, leading to confusion among consumers as well as food producers and legislators and to a lack of guidance for food producers. The authors prepared a pragmatic method and usable criteria to evaluate ingredients and foods to label them as ‘natural’. In this paper they describe the background, legislation, scope, methods used for qualification of ingredients and foods, the qualification of GMO Food and ingredients and an example to clarify the method. This pragmatic approach makes it easier for producers, retailers and consumers to share information on the term ‘natural’ in food labelling.

A geomechanical modeling study was conducted to investigate stability of major faults during past gas production and future underground gas storage operations in a depleted gas field in the Netherlands. The field experienced induced seismicity during gas production, which was most likely caused by the reactivation of an internal Central fault separating the two major reservoir blocks. A 3D field scale geomechanical finite element model of the gas field was developed with realistic representation of the structural geology and juxtaposition of various lithologies across the Central fault. The model was calibrated to match the subsidence data and the approximate location of the critically stressed, reactivated part of the fault in agreement with the seismological localization of the hypocenters of the past major seismic events. The model predicted a maximum shear slip of up to 2 cm associated with gas production. Additional, but a smaller, fault slip of up to 0.5 cm could be expected during the subsequent phase of cushion gas injection. During annual cycles of gas injection and production, the Central fault is not critically stressed and the predicted stress changes lie in the elastic region. Although the fault slip is unlikely, continuous monitoring of induced seismicity is essential.

It is increasingly apparent that the microbial ecosystems in the mammalian gastrointestinal tract play an intricate role in health and disease. There is a growing interest in the development of targeted strategies for modulating health through the modification of these microbiota. Ecologists are faced with the challenge of understanding the structure and function of ecosystems, the component parts of which interact with each other in complex and diffuse ways. The human gut microbiota, with its high species richness and diversity (up to 1000 bacterial species per individual) including members of all three domains of life, situated in the dynamic environment of the gastrointestinal tract, is probably among the most complex ecosystems on this planet. In order to elucidate the mechanistic foundations, and physiological significance, of beneficial or pathogenic relationships between the gut microbiota and their hosts, researchers require tractable model ecosystems that allow to recapitulate and investigate host-microbe and microbe-microbe interactions. This review discusses ex vivo gastrointestinal models systems that can be used to gain mechanistic insights into the emergent properties of the host-microbial superorganism. © 2013 Elsevier Ltd. All rights reserved.

Microbiota plays a role in the release and absorption of nutrients from feed components, thereby affecting digesta composition and moisture content of the excreta. The objective of the current study was to determine the effects of 5 different diets varying in ingredients (medium-chain fatty acids, nonstarch polysaccharides, and starch) on the microbiota composition of ileal digesta of broiler chickens and excreta DM content. Each treatment was repeated 6 times in cages each containing 18 Ross 308 broilers, with growth performance measured from 0 to 34 d of age and excreta DM and ileal microbiota composition analyzed at 34 d of age. Microbiota composition was evaluated using a novel ribosomal RNA microarray technology containing 370 different probes covering various genera, groups of microbial species, and individual species of the chicken gut microbiota, of which 321 had a signal above the background threshold. Replacing part of the animal fat and soybean oil in the wheat-based diet with mediumchain fatty acids (MCFA; 0.3% C10 and 2.7% C12) improved feed efficiency compared with the other dietary treatments. This coincided with a suppression of gram-positive bacteria belonging to the phylum of the Firmicutes, including Lactobacillus species, and species belonging to the family of the Enterococcaceae and Micrococcaceae, whereas the gram-negative bacteria belonging to the family of the Enterobacteriaceae were promoted. None of the other diets used in the present study notably changed the ileal digesta bacteria composition. Excreta DM content was not affected by dietary treatment. The variation between individual birds per dietary treatment was more pronounced than variation caused by feed composition, with the exception of the digesta microbiota of the birds fed the MCFA diet. It is concluded that a diet with MCFA significantly changes the ileal microbiota composition, whereas the effect of the other diets on the composition of the microbiota and excreta DM content is small in broiler chickens. © 2013 Poultry Science Association Inc.

This study examines the role of horizontal grid resolution on the performance of the regional three dimensional chemical transport model (CTM) PMCAMx. Two cases were investigated. First, the model was applied over the Northeastern United States with grid resolutions of 36 and 12 km during both a summer and a winter period. In this case the emission inventory was simply interpolated from the low resolution version. In the second case a multi-scale 36/4 km grid resolution is used over Western Europe with high resolution (4 × 4 km) emissions. The improvement in model predictions with the fine grid is modest during the summer and more significant during wintertime at both domains. Major differences are predicted mostly for primary (organics and black carbon (BC)) rather than secondary (e.g. sulfate) species. The use of high grid resolution decreases the bias for BC concentration by more than 30% in the Northeastern US during wintertime. In the Megacity of Paris, using high resolution emissions and a 4-km grid decreases the fractional bias for organic aerosol (OA) from 80% to 60% in the city center and produces much larger spatial concentration gradients in the domain as compared to the emissions being interpolated. These results suggest that future work should combine high grid resolution with high resolution emissions, taking into consideration, though, that the grid resolution is not currently the major source of the discrepancies between model predictions and observations. © 2012 Elsevier Ltd.

Urban search and rescue (USAR) missions are unique and unpredictable. Communication and coordination is difficult, with high-level actors (e.g. mission commander) lacking local terrain knowledge while low-level actors (e.g. robot operators, in-field rescuers) lack global situation awareness (SA). In-field actors have high mobility and a direct view of the field, but so far they could communicate this SA almost exclusively through radio. Unfortunately, words are imprecise and unorganized, and thus not easily analyzable and retrievable. As part of the NIFTi1 project, we developed mobile applications to help in-field actors share their SA with the rescue team. We also performed high-fidelity USAR simulations and missions at fire fighting training sites and disaster areas. We discovered a need for persistent geo-localized information and propose a novel system architecture that integrates pictures taken from robots and from in-field rescuers into the existing systems at the command post.

The successful surface waves retrieval in solid-Earth seismology using long-time correlations and subsequent tomographic images of the crust have sparked interest in extraction of subsurface information from noise in the exploration seismology. Subsurface information in exploration seismology is usually derived from body-wave reflections > 1 Hz, which is challenging for utilization of ambient noise. We use 11 h of noise recorded in the Sirte basin, Libya. First, we study the characteristics of the noise. We show that the bulk of the noise is composed of surface waves at frequencies below 6 Hz. Some noise panels contain nearly vertically traveling events. We further characterize these events using a beamforming algorithm. From the beamforming, we conclude that these events represent body-wave arrivals with a fairly rich azimuthal distribution. Having body-wave arrivals in the noise is a prerequisite for body-wave reflections retrieval. We crosscorrelate and sum the recorded ambient-noise panels to retrieve common-source gathers, following two approaches - using all the noise and using only noise panels containing body-wave arrivals likely to contribute to the reflections retrieval. Comparing the retrieved gathers with active seismic data, we show that the two-way traveltimes at short offsets of several retrieved events coincide with those of reflections in the active data and thus correspond to apexes of reflections. We then compare retrieved stacked sections of the subsurface from both approaches with the active-data stacked section and show that the reflectors are consistent along a line. The results from the second approach exhibit the reflectors better. © 2013. American Geophysical Union. All Rights Reserved.

Background: Type 2 diabetes mellitus (T2D) is a common age-related disease, and is a major health concern, particularly in developed countries where the population is aging, including Europe. The multi-scale immune system simulator for the onset of type 2 diabetes (MISSION-T2D) is a European Union-funded project that aims to develop and validate an integrated, multilevel, and patient-specific model, incorporating genetic, metabolic, and nutritional data for the simulation and prediction of metabolic and inflammatory processes in the onset and progression of T2D. The project will ultimately provide a tool for diagnosis and clinical decision making that can estimate the risk of developing T2D and predict its progression in response to possible therapies. Recent data showed that T2D and its complications, specifically in the heart, kidney, retina, and feet, should be considered a systemic disease that is sustained by a pervasive, metabolically-driven state of inflammation. Accordingly, there is an urgent need (1) to understand the complex mechanisms underpinning the onset of this disease, and (2) to identify early patient-specific diagnostic parameters and related inflammatory indicators. Objective: We aim to accomplish this mission by setting up a multi-scale model to study the systemic interactions of the biological mechanisms involved in response to a variety of nutritional and metabolic stimuli and stressors. Methods: Specifically, we will be studying the biological mechanisms of immunological/inflammatory processes, energy intake/expenditure ratio, and cell cycle rate. The overall architecture of the model will exploit an already established immune system simulator as well as several discrete and continuous mathematical methods for modeling of the processes critically involved in the onset and progression of T2D. We aim to validate the predictions of our models using actual biological and clinical data. Results: This study was initiated in March 2013 and is expected to be completed by February 2016. Conclusions: MISSION-T2D aims to pave the way for translating validated multilevel immune-metabolic models into the clinical setting of T2D. This approach will eventually generate predictive biomarkers for this disease from the integration of clinical data with metabolic, nutritional, immune/inflammatory, genetic, and gut microbiota profiles. Eventually, it should prove possible to translate these into cost-effective and mobile-based diagnostic tools. © Filippo Castiglione, Paolo Tieri, Albert De Graaf, Claudio Franceschi, Pietro Liò, Ben Van Ommen, Claudia Mazzà, Alexander Tuchel, Massimo Bernaschi, Clare Samson, Teresa Colombo, Gastone C Castellani, Miriam Capri, Paolo Garagnani, Stefano Salvioli, Viet Anh Nguyen, Ivana Bobeldijk-Pastorova, Shaji Krishnan, Aurelio Cappozzo, Massimo Sacchetti, Micaela Morettini, Marc Ernst.

Introduction. Diagnosis of exposure to organophosphate anti-cholinesterases can play a pivotal role in case of a military or terrorist attack with such agents, e.g. for forensic or medical purposes. Current methods used for assessment of such exposure include the determination of cholinesterase activity and analysis of urinary metabolites. Since these biomarkers are mostly short-lived, there is a need for additional and more persistent biomarkers. Methods. We here present various diagnostic methods for detection of OP exposure, based on analysis of covalent adducts to butyrylcholinesterase (BuChE) and albumin. The fluoride reactivation method is based on the regeneration of the organophosphofluoridate upon incubation of a plasma sample with fluoride ions. The regenerated organophosphofluoridate can be detected with GC-MS (SIM) to ensure high sensitivity, or by using GCxGC – TOF MS, which has the advantage that selectivity and sensitivity are combined with identification using full mass spectra. Phosphylated BuChE can also be analyzed with liquid chromatography – tandem mass spectrometry by detection of a phosphylated nonapeptide resulting after pepsin digestion of BuChE. This method is important in case the adducts are refractory towards fluoride reactivation, as is for ‘aged’ adducts. Covalent adducts formed with albumin can easily be detected after pronase or pepsin digestion of the protein. Results. The utility of the abovementioned methods is illustrated through the analysis of plasma samples from intoxicated patients taken 33-49 days after exposure to the organophosphate pesticides chlorpyrifos and diazinon. Using the fluoride reactivation method, the phosphyl moiety was released from the protein, resulting in O,O-diethyl phosphorofluorothioate and its oxon analog, as evidenced by GC-MS analysis. Analysis of pepsin-digested BuChE revealed that only the aged adduct of the oxon-analog of the pesticides was detected as the FGES*AGAAS peptide, with S* representing the serine residue modified with an (aged) ethyl phosphoric ester moiety. Upon isolation and digestion of albumin with pronase, tyrosine modified with a O,O-diethylphophorothiono moiety could be detected, stemming from the thioate form of the pesticides. Conclusions. Several adduct-based methods have been developed for unequivocal, retrospective exposure assessment to organophosphates. The high utility of these methods

Soil nitrogen (N) budgets are used in a global, distributed flow-path model with 0.5° &times 0.5° resolution, representing denitrification and N2O emissions from soils, groundwater and riparian zones for the period 1900-2000 and scenarios for the period 2000-2050 based on the Millennium Ecosystem Assessment. Total agricultural and natural N inputs from N fertilizers, animal manure, biological N2 fixation and atmospheric N deposition increased from 155 to 345 Tg N yr-1 (Tg 1/4 teragram; 1 Tg 1/4 1012 g) between 1900 and 2000. Depending on the scenario, inputs are estimated to further increase to 408-510 Tg N yr21 by 2050. In the period 1900-2000, the soil N budget surplus (inputs minus withdrawal by plants) increased from 118 to 202 Tg yr-1, and this may remain stable or further increase to 275 Tg yr-1 by 2050, depending on the scenario. N2 production from denitrification increased from 52 to 96 Tg yr21 between 1900 and 2000, and N2O-N emissions from 10 to 12 Tg N yr-1. The scenarios foresee a further increase to 142 Tg N2-N and 16 Tg N2O-N yr-1 by 2050. Our results indicate that riparian buffer zones are an important source of N2O contributing an estimated 0.9 Tg N2O-N yr-1 in 2000. Soils are key sites for denitrification and are much more important than groundwater and riparian zones in controlling the N flow to rivers and the oceans. © 2013 The Author(s) Published by the Royal Society. All rights reserved.

Over the past twenty years, the number of mobile phone subscriptions has risen from 12 million to more than six billion. 75% of the world has access to a mobile phone and the developing world is now more mobile than the developed world. However, the growth of non-voice services on these mobiles for the Base of the Pyramid users is still at the start of its growth curve. Drawing from a study of 50 case studies in financial services, this paper examines how the innovation for financial services was organized using a framework of six elements. It shows the importance of government support (both as regulator and as (launching) customer), the need for trust delivered by expansion of existing strong brands, embeddedness by using local agents and capacity building of these agents and the customers. © 2013 The Authors.

Fibrillar and spherical aggregates were prepared from whey protein isolate (WPI). These aggregates were thiolated to a substantial degree to observe any impact on functionality. Sulfur-containing groups were introduced on these aggregates which could be converted to thiol groups by deblocking. Changes on a molecular and microstructural level were studied using tryptophan fluorescence, transmission electron microscopy, and particle size analysis. The average size (nm) of spherical aggregates increased from 38 to 68 nm (blocked variant) and 106 nm (deblocked variant) after thiolation, whereas the structure of fibrillar aggregates was not affected. Subsequently, gels containing these different aggregates were prepared. Rheological measurements showed that thiolation decreased the gelation concentration and increased gel strength for both WPI fibrillar and spherical aggregates. This effect was more pronounced upon thiolation of preformed fibrillar aggregates. The findings suggest that thiolation at a protein aggregate level is a promising strategy to increase gelation efficiency. © 2013 American Chemical Society.

This work presents an approach to compute dryer energy efficiency using air flowrate step responses and establish a link between drying energy efficiency and process controllability. The approach is based on the temperature drop between the dryer inlet and outlet air under adiabatic conditions and so decouples water evaporation from heat loss and product heating effects on dryer temperature drop. As such, the computation is accurate even for dryers with significant heat losses where the traditional use of actual temperature drop measurements is grossly inaccurate. The approach is tested and verified on two experimental case studies involving significant heat losses: the first, a continuous fluidized-bed dryer (from literature); the second, a conventional and zeolite wheel-assisted batch dryer designed in the current study. © 2013 Copyright Taylor and Francis Group, LLC.

This guide provides a major revision and update of the lithostratigraphy of the Rogaland Group for the Norwegian North Sea. An abundance of recent well and seismic data sheds new light on lithology, biostratigraphy, provenance, geographic distribution and terminology of all Rogaland rock units, used widely in the search for oil and gas. While finer siliciclastic units largely remain as previously defined, previous sandstone/siltstone formations and one (reworked) chalky unit are now re-defined. These lithostratigraphic units are local sediment bodies of a lithology different from the surrounding and embracing formation. Hence, these lithostratigraphic units are members in the formal stratigraphical hierarchy. With the new definitions and re-definitions the Rogaland Group now consists of four formations and 15 members, which span the stratigraphic interval from lower Paleocene to lower Eocene. The revisions concerning the sandstone bodies are of four different types: – Re-definition from formations to members – Re-definition of lithological criteria – Introduction of members long used already offshore England and Denmark – Definition of new members For those practicing geologists not familiar with historic precedence, an important ʻsine qua nonʼ in (litho-) stratigraphy, it should be pointed out that 8 out of 15 members discussed here have been predefined in literature dealing with the UK and Danish sectors of the North Sea. The present study thus updates the Norwegian lithostratigraphic bulletins of the nineteen eighties for its offshore area. The internet site www.nhm2.uio.no/ norlex provides an interactive digital version of this study, with links to well data, biozonations and core archives relevant to the Rogaland Group.

Bile acid sequestrants (BASs) are cholesterollowering drugs that also affect hyperglycemia. The mechanism by which BASs exert these and other metabolic effects beyond cholesterol lowering remains poorly understood. The present study aimed to investigate the effects of a BAS, colestilan, on body weight, energy expenditure, and glucose and lipid metabolism and its mechanisms of action in high-fat-fed hyperlipidemic APOE*3 Leiden (E3L) transgenic mice. Mildly insulin-resistant E3L mice were fed a high-fat diet with or without 1.5% colestilan for 8 weeks. Colestilan treatment decreased body weight, visceral and subcutaneous fat, and plasma cholesterol and triglyceride levels but increased food intake. Blood glucose and plasma insulin levels were decreased, and hyperinsulinemic-euglycemic clamp analysis demonstrated improved insulin sensitivity, particularly in peripheral tissues. In addition, colestilan decreased energy expenditure and physical activity, whereas it increased the respiratory exchange ratio, indicating that colestilan induced carbohydrate catabolism. Moreover, kinetic analysis revealed that colestilan increased [ 3 H]NEFA incorporation in biliary cholesterol and phospholipids and increased fecal lipid excretion. Gene expression analysis in liver, fat, and muscle supported the above findings. In summary, colestilan decreases weight gain and improves peripheral insulin sensitivity in high-fat-fed E3L mice by enhanced NEFA incorporation in biliary lipids and increased fecal lipid excretion. Copyright © 2013 by the American Society for Biochemistry and Molecular Biology, Inc.

This thesis presents an algorithm that can reduce the estimation errors made with the DriftLessTM bias estimation technique. The algorithm utilizes the autocorrelation function to detect the presence of errors, and a minimization function to reduce these errors. The algorithm has been validated with simulations. A sensor signal contains noise, bias and the measurand signals. Bias is a non-constant, non-random, additive error on the measurement signal of a sensor. DriftLessTMuses two sensors that measure a vectorial quantity e.g. accelerometers or gyroscopes. To estimate the bias these sensors are rotated mechanically, physically with respect to each other. DriftLessTM rotates the sensor signals back numerically to make both sensors measure the same measurand virtually. Then these signals are subtracted from each other which results in a signal that is ideally independent of the measurand because the two measurands measured by the sensors will cancel each other out. The newly created signal still contains the biases. From this DriftLessTMsignal vector the bias is estimated. In reality the rotations are known with finite accuracy and precision. This causes an estimation error because a part of the measurand signal remains in the DriftLessTM signal vector. An estimate of the magnitude of the estimation error can be made by calculating the sample autocorrelation of each entry of the DriftLessTMsignal vector after correcting for the bias. The algorithm works by minimizing the autocorrelation of the bias corrected DriftLessTMsignal vector with respect to a numerical rotation matrix. Then the bias is re-estimated with the numerical rotation matrix that solved the previous minimization. Solving for a numerical rotation matrix and re-estimating the biases is repeated until convergence of the bias estimation. The algorithm increases the accuracy and precision of the DriftLessTMbias estimation method under certain circumstances. The increase in accuracy and precision is higher as the cross correlation of the measurand and the bias signals decreases, the power of the noise increases, the misalignments are greater and there is more time available for the computations of the algorithm.

The development and implementation of effective systems to identify vulnerabilities in food chains to chemical and microbiological contaminants must take account of consumer priorities and preferences. The present investigation attempted to understand consumer perceptions associated with chemical and microbiological contaminants in four specific food chains (drinking water, farmed salmon, chicken and milk powder). To this end, ten focus group discussions were held in five different countries (Poland, Ireland, the Netherlands, France and Brazil). Consumers expressed higher concerns about chemical, as compared with microbial, contaminants. Chemical contaminants were more strongly associated with the potential for severe consequences, long-term effects and lack of personal control. Traceability was considered by consumers as a useful tool that offers the potential to improve consumer confidence in food safety. © 2011 Blackwell Publishing Ltd.

Shallow gas accumulations in the Plio-Pleistocene Eridanos Delta deposits in the Dutch offshore are a valuable additional hydrocarbon source. Ongoing studies focus on better understanding the shallow gas play. The focus of this paper is on increasing the understanding of the origin of the shallow gas. For that purpose we used integrated modeling approaches to assess the timing and spatial distribution of biogenic gas generation. It was found that intra-Eridanos Delta deposits are the most probable source of biogenic gas and that these biogenic sources are at maximum depth and temperature today. Detailed glacial-interglacial temperature fluctuations do not have a large influence on the history of the optimum window for biogenic gas generation, i.e. on start of biogenic gas generation. In the oldest and deepest buried units of the Delta, biogenic gas generation started already at the beginning of the Pleistocene, while the youngest units have not yet reached the optimum window for biogenic gas generation. Copyright © (2012) by the European Association of Geoscientists & Engineers All rights reserved.