Immunoglobulin 1 (IgG1) Fc-glycosylation profiling of anti-citrullinated peptide antibodies from human serum

In several autoimmune disorders, including rheumatoid arthritis (RA), autoantibodies are thought to be the driving force of pathogenicity. Glycosylation of the Fc-part of human Igs is known to modulate biological activity. Hitherto, glycosylation of human IgG-Fc has been analyzed predominantly at the level of total serum IgG, revealing reduced galactosylation in RA. Given the pathogenic relevance of autoantibodies in RA, we wished, in the present study, to address the question whether distinct Fc-glycosylation features are observable at the level of antigen-specific IgG subpopulations. For this purpose, we have developed a method for the microscale purification and Fc-glycosylation analysis of anti-citrullinated peptide antibodies (ACPA). ACPA represent a group of autoantibodies that occur with unique specificity in RA patients. Their presence is associated with increased inflammatory disease activity and rapid joint destruction. Results indicate that ACPA of the IgG1 subclass vary considerably from total serum IgG1 with respect to Fc-galactosylation, with galactosylation being higher on ACPA than on serum IgG1 for some patients, while other patients show higher galactosylation on serum IgG1 than on ACPA. Using this method, studies can be performed on the biological and clinical relevance of ACPA glycosylation within RA patient cohorts.     

Generation of time delays: simplified models of intracellular signalling in cerebellar Purkinje cells

In many neuronal systems, information is encoded in temporal spike patterns. The recognition and storage of temporal patterns requires the generation and modulation of time delays between inputs and outputs. In cerebellar Purkinje cells, stimulation of metabotropic glutamate receptors (mGluRs) results in a delayed calcium and voltage response that has been implicated in classical conditioning and temporal pattern recognition. Here, we analyse and simplify a complex model of the intracellular signalling network that has been proposed as a substrate for this delayed response. We systematically simplify the original model, present a minimal model of time delay generation, and show that a delayed response can be produced by the combination of negative feedback and autocatalysis, without any intervening signalling steps that would contribute additive delays. The minimal model is analysed using phase plane methods, and classified as an excitable system. We discuss the implication of excitability for computations performed by intracellular signalling networks in general.     

The use of interorganisational ICT in United States construction projects

In recent research, the use of ICT applications in real time construction projects have been documented and analysed extensively. However, there is a need for identifying and analysing in-depth the mechanisms influencing the use of interorganisational ICT applications and for solutions to eliminate potential barriers to the successful use of ICT. A model is developed providing insights in these mechanisms. Based on this model and expert interviews in the United States construction industry directions for solutions to barriers to the successful use of interorganisational ICT — document management applications, workflow management applications, and product modelling applications — in construction projects are formulated. These solutions focus on stimulating the personal motivation to use ICT, the external motivation to use this technology, and facilitating conditions in terms of knowledge and skills and acting opportunities to use ICT.     

Improved creaminess of low-fat yoghurt: The impact of amylomaltase-treated starch domains

Amylomaltase-treated starch (ATS) is an excellent creaminess enhancer in yoghurt. Small amounts of ATS raised the creaminess perception of low-fat yoghurt (1.5%) to that of full-fat yoghurt (5%). In this way, a reduction in fat-related energy value could be achieved from 45 to 21.5 kcal/100 g product. The functionality of ATS in set yoghurt resulted from discrete domains of ATS that resemble the microstructural behaviour of fat particles. The microstructure of the yoghurt is dominated by the protein and the ATS domains are enclosed in or bound to this protein network. The perceived creaminess resulted from in-mouth melting of these ATS domains due to a combined effect of their physical melting and hydrolysis by amylase present in the saliva.     

The effect of dendritic topology on firing patterns in model neurons

Neuronal firing patterns are influenced by both membrane properties and dendritic morphology. Distinguishing two sources of morphological variability-metrics and topology-we investigate the extent to which model neurons that have the same metrical and membrane properties can still produce different firing patterns as a result of differences in dendritic topology. Within a set of dendritic trees that have the same number of terminal segments and the same total dendritic length, we show that firing frequency strongly correlates with topology as expressed by the mean dendritic path length. The effect of dendritic topology on firing frequency is bigger for trees with equal segment diameters than for trees whose segment diameters obey Rall's 3/2 power law. If active dendritic channels are present, dendritic topology influences not only firing frequency but also type of firing (regular, bursting).     

Size, differentiation and the performance of Dutch daily newspapers

We demonstrate how organizational ecology can contribute tostrategic management and managerial practice by using resource-partitioningtheory to make predictions with respect to: (i) the short-runperformance (i.e. growth and profitability) consequences ofbroad (generalist) vis-à-vis focus (specialist) strategies;(ii) the detrimental performance implications of a particularinstance of being strategically ‘stuck in the middle’;and (iii) the performance consequences of organizational sizedifferences. We hypothesize that these effects depend on theposition of the organization in resource space. These predictionsare tested by estimating models of growth and profitability,using data on Dutch generalist (national) and specialist (regional)newspapers from 1968 to 1994. The findings show that resource-partitioningtheory provides a dynamic backbone to understand the performanceconsequences of different generic strategies and organizationalsize positions.     

On-line coupling of high-performance liquid chromatography to a continuous-flow enzyme assay based on electrospray ionization mass spectrometry

Liquid chromatography (LC) was coupled on-line to a continuous-flow enzymatic assay using electrospray ionization mass spectrometry (ESI-MS) as readout for the screening of enzyme inhibitors in complex samples. Inhibitors were detected by changes in the concentration of the enzymatic reaction products, indicating the inhibition of enzymatic activity. The molecular masses of the inhibitors were determined with high certainty by using retention time matching and peak shape comparison. Due to the high matching accuracy, baseline separation of coeluting analytes was not necessary in order to identify the correct masses of the bioactive compounds. The continuous-flow system was successfully applied for the screening of complex samples, such as natural extracts. For a red clover extract, detection limits of 0.3-0.8 micromol/L were obtained. System validation was performed by determining the IC(50) values of four inhibitors in the flow-injection mode. The IC(50) values were in the 0.11-5.6 micromol/L range and correspond closely to data obtained by microtiter plate assays. Detection limits were in the range of 0.018-0.35 micromol/L in the flow-injection mode, and 0.075-0.75 micromol/L in the LC mode. These values are well below the typical compound concentrations (1-10 micromol/L) used in high-throughput screening. Together with an interday precision of 12.6%, these results demonstrate the applicability of the system for bioactivity screening of complex mixtures, generating both chemical and biological information on bioactive compounds in a single run.     

Casein micelles as a vehicle for iron fortification of foods

Casein micelles were loaded with iron to create a dispersible delivery format for insoluble iron by exposing milk at chilled temperatures to a high concentration of soluble iron (up to 20 mmol kg⁻¹ ferrous and ferric chloride). The loading was maximised by applying a pH-cycle to the fortified milk by means of carbonation. Upon acidification of fortified milk, no release of iron was observed, except at the highest concentration. Changes in the buffering capacity as a function of pH confirmed the formation of colloidal iron phosphates. Overall, most properties of the micelles did not change: hydration, protein distribution between soluble and colloidal phase remained constant, but zeta potential decreased slightly and curd formation upon renneting became much slower. The renneting behaviour could be improved by carbonation or storage at 30 °C for a day. Iron-fortified milk samples were stable under heating, except when fortification was achieved by means of 20 mmol kg⁻¹ ferric chloride. The most obvious difference of iron-fortified milk is its appearance: samples fortified with ferrous chloride were darker than control, whereas samples fortified with ferric chloride were more red/yellow.     

Corporate Water Footprint Accounting and Impact Assessment: The Case of the Water Footprint of a Sugar-Containing Carbonated Beverage

All water use in the world is ultimately linked to final consumption by consumers. It is therefore interesting to know the specific water requirements of various consumer goods, particularly the water-intensive ones. This information is relevant not only for consumers, but also for food processors, retailers, and traders. The objective of this paper is to carry out a pilot study on water footprint accounting and impact assessment for a hypothetical sugar-containing carbonated beverage in a 0.5 l PET-bottle produced in a hypothetical factory that takes its sugar alternatively from sugar beet, sugar cane and high fructose maize syrup and from different countries. The composition of the beverage and the characteristics of the factory are hypothetical but realistic. The data assumed have been inspired by a real case. This paper does not only look at the water footprint of the ingredients of the beverage, but also at the water footprint of the bottle, other packaging materials and construction materials, paper and energy used in the factory. Although most companies focus on their own operational performance, this paper shows that it is important to consider freshwater usage along the supply chain. The water footprint of the beverage studied has a water footprint of 150 to 300 l of water per 0.5 l bottle, of which 99.7–99.8% refers to the supply chain. The study also shows that agricultural ingredients that constitute only a small fraction in weight of the final product have the biggest share at the total water footprint of a product.