CD40/APC-specific antibodies with three T-cell epitopes loaded in the constant domains induce CD4+ T-cell responses

CD4+ T lymphocytes play a central role in the orchestration and maintenance of the adaptive immune response. Targeting of antigen to antigen presenting cells (APCs) increases peptide loading of major histocompatibility complex (MHC) class II molecules and CD4+ T-cell activation. APCs have been targeted by APC-specific recombinant antibodies (rAbs) with single T-cell epitopes integrated in the constant region of the heavy chain (C(H)). However, the strategy may be improved if several T-cell epitopes could be delivered simultaneously by one rAb. We here demonstrate that a single rAb can be loaded with multiple identical or different T-cell epitopes, integrated as loops between β-strands in C(H) domains. One epitope was inserted in C(H)1, while two were placed in C(H)2 of IgG. T-cell proliferation assays showed that all three peptides were excised from loops and presented on MHC class II to T-cells. Induction of T-cell activation by each epitope in the multi-peptide rAb was as good, or even better, than that elicited by corresponding single-peptide rAbs. Furthermore, following DNA vaccination of mice with plasmids that encode CD40-specific rAbs loaded with either one or three peptides, T-cell responses were induced. Thus, integration of multiple epitopes in C(H) region loops of APC-specific rAbs is feasible and may be utilized in design of multi-vaccines.     

Low-energy and high-fibre liver paté processed using potato pulp

Danish liver paté was processed using water and potato pulp from the processing of potato flour as fat substitute. The spreading characteristics of the patties were improved using Citrem as emulsifier and satisfactory texture characteristics were obtained using fibre soaked in ten times its own weight. Colour and texture were measured using a Minolta Chroma Meter for determination of the Hunter Lab values and a Farnell Quality Test System for measurement of texture characteristics. Baking loss decreased at low water to fibre ratios (water:fibre) from 3 to 4 and then increased with increasing water:fibre. No clear relationships were observed between the water:fibre and yellowness, lightness or redness. Three factors extracted by factor analysis depended especially on hardness–water:fibre, adhesion–water:fibre and cohesiveness–water:fibre, respectively. Generally, all textural characteristics were inversely correlated to water:fibre. Hardness 1 and 2, deformation energy 1 and 2, gumminess and modulus decreased in similar fashion with respect to to water:fibre, whereas adhesiveness, cohesiveness and springiness varied in relation to water:fibre. The effect of emulsifier on the texture was less than that of water:fibre. The patties with water:fibre of 10–11 were easy to spread on bread and had very attractive sensory properties. The increased substitution of fat by potato pulp and water decreased the energy content by 27% and increased the yield of baked patties by 33% (w/w).     

Betula alba Bark Extract and Empetrum nigrum Fruit Juice,a Natural Alternative to Niacinamide for Skin Barrier Benefits

The Scandinavian region is home to a unique biome with endemic plant species. The aim of this study was to explore this natural diversity and identify plant extracts providing positive skin barrier effects. Six plant extracts were identified as starting material. Following biochemical screening, two candidates outperformed the rest: Betula alba (BA) and Empetrum nigrum (EN). Quantitative PCR analysis showed that BA and EN upregulated barrier genes, when used individually and in combination. Betula alba increased AQP3 and OCLN protein expression, something niacinamide was incapable of. Additionally, the skin barrier was strengthened, evidenced by inhibition of KLK5 and hyaluronidase and showed strong antioxidant and anti-inflammatory activity through DPPH and COX2 inhibition, respectively. A first split-face clinical study was conducted using the combination of extracts versus placebo. There was a significantly better skin restructuring effect and corneocyte cohesion on the side treated with combined extracts. A second split-face clinical study assessed the combined extracts versus 3% niacinamide. Significant variations in skin hydration and TEWL were observed in favor of the extract treated side. In conclusion, we identified a natural alternative to niacinamide for improving skin barrier health, in Scandinavian plant extracts, which yield strong performance, but at a lower concentration.     

Immunoaffinity chromatography purification and characterisation of pea trypsin inhibitors

Trypsin inhibitors from pea (Pisum sativum (L) cultivar Progreta grown in Denmark) have been isolated and shown to consist of at least nine pea proteinase inhibitors (PPI) with inhibitor activity toward both trypsin and chymotrypsin. The isoelectric points of the inhibitors were in the range 4.9–7.8. From this PPI mixture at least four inhibitors were isolated by immunoaflinity chromatography on a column containing immobilised monoclonal antibody (mAb) with inhibitor specificity. The PPI isolated by immunoaffinity chromatography were further separated by HPLC, and subsequent SDS-PAGE analysis showed molecular weights for four of the PPI in the range 11.3–14.2 kD. Their pI were determined by isoelectric focusing, mAbs were used for immunochemical characterisation and their amino acid composition showed a high (14.7–21%) content of cysteine. Tryptophan was not present in any of the isolated PPI. The data now obtained support the resemblance of PPI with inhibitors of the Bowman-Birk class and the differences in immunochemical properties of the various PPI indicate that pea has at least two gene loci coding for the inhibitors.     

USE OF FLOW CYTOMETRY FOR RAPID ESTIMATION OF INTRACELLULAR EVENTS IN BREWING YEASTS

A flow cytometric method based on staining with fluorescein diacetate (FDA) for rapid measurement of the fluorescence intensity (FI) of individual brewing yeast cells is reported. Distinct changes in the FI of yeast populations during the course of both laboratory scale and brewery fermentations were observed. The changes followed a characteristic pattern defined as the FI-profile. The FI-profile was related to the rate of attenuation as well as changes in yeast glycogen and ergosterol contents. The results obtained suggest that flow cytometry can be applied for estimation of intracellular events in brewing yeasts and prediction of their performance.     

MMPI-2 random responding indices: Validation using a self-report methodology.

Investigated the validity of F, back F, and variable response inconsistency indices of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) for detection of self-reported partially random responding in several samples, including college students, community volunteers, and job applicants. Substantial numbers of Ss admitted to providing some random responses, with 29–60% of the various samples acknowledging an average of 12–38 random responses in their MMPI-2 answers. For all groups, with the expected exception of job applicants, the appropriate MMPI-2 validity indices were reliably and positively correlated with self-estimates of random responses. Available data suggested that random responding occurred most commonly toward the end of the test, although significant numbers indicated that they had scattered their random responses throughout the test. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Reduced combustion efficiency of chlorinated compounds,resulting in higher yields of carbon monoxide

A series of experiments involving the combustion of inherently fire-retarded compounds has been performed using the DIN 53 436 furnace. Six chlorinated compounds comprising four pesticides, an organic solvent, and polyvinyl chloride (PVC) have been investigated with the focus on the combustion efficiency expressed as the CO₂/CO ratio in the fire effluent. It was found that increased chloride content in a compound resulted in decreased CO₂/CO ratio. The correlation was found to be $ \sqrt {{\rm CO}_{\rm 2} /{\rm CO}} = 0.61 \times ({\rm Cl/C)}^{- {\rm 1}} + 0.13 $. The results indicate higher concentrations of CO in the fire effluent from chlorinated compounds than from wood. Together with a release of HCI this could result in an increased toxic potency. The overall toxic hazard may or may not be increased, since (inherently) fire-retarded materials tend to have an improved fire performance.     

Control of Salmonella in food related environments by chemical disinfection

Salmonella may be transferred to food through cross-contamination during processing and preparation. To minimise the risk of cross-contamination, proper cleaning and disinfection is essential for the food industry. Recently, disinfection of areas for preparation and storage of food has also gained increased popularity in households. There is a range of disinfectants available with different properties and usage areas, and care must be taken to choose the proper disinfectant for the specific application.There are many methods for testing the antimicrobial effect of disinfectants. To evaluate whether a disinfectant will be effective in practical settings, the test method should model real-life situations. Most disinfectants are effective against Salmonella at recommended user concentration in suspension tests. However, a number of factors may reduce the biocidal effect of disinfectants under practical conditions. This include properties of the surface to be disinfected, presence of soiling on the surface, the physiological state of the bacteria exposed to disinfection, including bacteria embedded in biofilms, and the effects of other stresses (e.g. desiccation, starvation and temperature).Here we review the effects of disinfectants used in food related areas in industries and in households against Salmonella. A general overview is given for disinfectants in use and methods used to evaluate effects. Effects of disinfectants against Salmonella in suspension and on surfaces, including biofilms, are presented and compared. Novel control strategies such as use of electrolysed water, antimicrobial surfaces, and anti-biofilm compounds are also covered. Finally, we review the ability of Salmonella to gain reduced susceptibility to disinfectants through adaptation and other physiological responses like biofilm formation.     

Structure of pectin in relation to abnormal hardness after cooking in pre-peeled, cool-stored potatoes

Unacceptable hard tissue can be detected in pre-peeled potatoes after cooking. Pectin methyl esterase (PME) has been suspected to facilitate formation of Ca-bridges between pectin molecules by deesterification of the uronic acids. But this has also been questioned, due to the low storage temperature of the peeled tubers, optimum of PME being 50-70 °C. Hardness in tubers was induced by several packaging methods. Annual and seasonal variations were observed. In the work, analysis of nonstarch polysaccharides (NSP) are considered representative of the pectin content. Analyses revealed an increase in insoluble NSP during storage after peeling and cooking, and a decrease in branching (arabinose, galactose) and degree of methyl esterification of total NSP, in both fresh and cooked tissue, dependant on storage period. Reduction of steric hindrance by debranching of the pectin molecules as well as a PME-induced increase of possible Ca-cross-linking sites may favor hardening of the tissue. The microscopy does not show any pectin changes until after cooking, but the chemical data do. This suggests that hardening, at least partly, is due to the removal of methyl ester groups, as well as galactose and arabinose in the side chains. The Ca-bridges can probably not be formed until sufficient calcium is released from the starch during gelatination.