Introducing a biosensor based technology for real-time biospecific interaction analysis

The peptido-leukotriene receptor(s) (PL) on the ferret isolated spleen strip have been characterised by functional studies using the naturally occurring leukotrienes (LTs), a range of structurally distinct PL antagonists, and by ligand binding studies. LTB4 (0.01-10 microM) was inactive on ferret spleen whereas LTC4, LTD4 and LTE4 produced concentration-related contractions with maximal responses, relative to noradrenaline, of 57% (EC50 0.28 microM), 60% (EC50 0.5 microM) and 7% respectively. The leukotriene responses were unaltered by L-serine borate, L-cysteine, indomethacin, phentolamine, propranolol, mepyramine, methysergide or atropine, suggesting that the peptido-leukotrienes were acting through distinct PL receptors. The PL1 antagonists, FPL 55712 (0.01-10 microM), ICI 198615 (10 microM), SK&F 104353 (10 microM) and MK541 (10 microM) were all inactive against LTC4- or LTD4-induced contractile responses. LTE4 was a partial agonist with respect to LTC4 and LTD4 with pKB values of 5.8 and 5.5 respectively. Nifedipine (0.1 microM) produced a rightward shift of the concentration-response curves to both LTC4 and LTD4 and depressed their maximal responses. An unacceptably high level of non-specific binding of [3H]LTD4 to membrane preparations of ferret spleen prevented characterisation of this receptor by ligand binding. These results suggest that the ferret spleen has a homogeneous population of a PL receptor type which is insensitive to existing PL1 receptor antagonists. The functional characteristics of this PL receptor type are similar to those of the PL2 receptor on other tissues. The absence of PL1 receptors on this tissue makes it particularly useful in identifying new and selective drug tools for the PL2 receptor.     

Delineation of the epitope recognized by an antibody specific for N-glycolylneuraminic acid-containing gangliosides

P3 is a mouse monoclonal antibody (mAb) that binds to several NeuGc-containing gangliosides. It also reacts with antigens expressed in human breast tumors (Vázquez et al. (1995) Hybridoma , 14, 551-556). In this work, the binding specificity of P3 has been characterized in more detail using a panel of glycolipids that included several disialylated gangliosides and several chemical derivatives of NeuGc-GM3. The carboxyl group and the nitrogen function of sialic acid were found to play important roles in the antibody binding, whereas the glycerol tail appears to be nonrelevant. Molecular modeling was used to analyze the binding data, including the finding that P3 selectively recognizes the internal NeuGc in GD3. For this purpose, conformational studies of GD3 were performed using molecular dynamics. It was concluded that sialic acid binds the P3 antibody through its upper face (the one on which the carboxyl group is exposed) and the C4-C5 side of the sugar ring, whereas none or very little contact between the galactose residue and the protein is evident. Conformational analysis of GD3 revealed that, despite the large flexibility of the NeuGcalpha8NeuGc linkage, the P3 binding epitope on the external sialic acid is not well exposed for any of the possible conformations this linkage can adopt, whereas the internal sialic acid presents the epitope in a proper way for several of these conformations. As a final result, a coherent picture of the epitope that fits the wide binding data was obtained.     

New results on the Fermi surface of β brass type alloys

De Haas-van Alphen frequencies not previously reported have been measured for -PdIn and -CuZn. The results for PdIn suggest a reinterpretation of some of the previous results and offer an explanation of a previously unexplained set of de Haas-van Alphen frequencies in PdIn and AgZn. A recent nonlocal pseudo-potential model by Taylor can account for the experimental results reported here as well as the earlier data.National Research Council of Canada Postdoctorate Fellow.     

Influence of early pH decline on calpain activity in porcine muscle

This study investigated the influence of post-mortem pH decline on calpain activity and myofibrillar degradation. From 80 pigs, 30 Longissimus dorsi (LD) muscles were selected on the basis of pH values at 3 h post-mortem and classified into groups of 10 as fast, intermediate and slow pH decline. The rate of pH decline early post-mortem differed between the three groups, but the ultimate pH values were similar at 24 h. Calpain activity and autolysis from 1 to 72 h post-mortem were determined using casein zymography and studied in relation to myofibrillar fragmentation.     

Internal doses of acrylamide and glycidamide in mice fed diets with low acrylamide contents

The formation of acrylamide during heating of certain foodstuffs constitutes a potential health hazard. The health risk assessment should be based on knowledge about the relation between dietary exposure to acrylamide and internal doses of acrylamide and its genotoxic metabolite glycidamide. The primary aim of this study in mice was to measure these relationships at low levels of acrylamide intake through the diet. A secondary aim was to clarify which extraction method should be used when analyzing acrylamide in food in order to obtain a correct measure of the acrylamide that is available for absorption. In the analysis procedure, alkaline extraction has earlier shown much higher measured acrylamide levels in certain foods compared to water extraction. In this subcronic study the administered diets were composed to give five levels of acrylamide intakes between 3 and 50 mug/kg body weight per day (calculated on figures obtained after water extraction). Internal doses of acrylamide and glycidamide were measured through hemoglobin (Hb)-adducts. The results showed linear relationships between the exposure of acrylamide and Hb-adduct levels from both acrylamide and glycidamide at these low exposure levels. The study also showed that the "extra" acrylamide measured with alkaline extraction does not correspond to bioavailable acrylamide.     

Role of charge and hydrophobicity in translocation of cell-penetrating peptides into Candida albicans cells

Although the interactions of cell-penetrating peptides (CPPs) with mammalian cells have been widely studied, much less is known about their interactions with fungal cells. To study how the properties of CPPs affect translocation into fungal cells, we designed variants of the peptides pVEC and SynB with altered levels of charge and hydrophobicity and evaluated the translocation of the variants into the important human fungal pathogen Candida albicans. Charge played a greater role in translocation efficacy of the peptides than hydrophobicity, with a higher net positive charge leading to higher level of translocation into C. albicans and a higher level of cytosolic localization. Hydrophobicity had little effect on translocation efficacy, but a low level of hydrophobicity did lead to increased vacuolar localization and an energy-dependent translocation mechanism. Our results suggest that CPPs can be designed for desired levels of cargo delivery into fungal cells and for desired translocation mechanisms.     

Characterization of iron(III) in organic soils using extended X-ray absorption fine structure spectroscopy

The distribution of different iron (Fe) species in soils, sediments, and surface waters has a large influence on the mobility and availability of Fe, other nutrients, and potentially toxic trace elements. However, the knowledge about the specific forms of Fe that occurs in these systems is limited, especially regarding associations of Fe with natural organic matter (NOM). In this study, extended X-ray absorption fine structure (EXAFS) spectroscopy was used to characterize Fe(III) in organic soils (pH 4.6-6.0) with varying natural Fe content. The EXAFS data were subjected to wavelet transform analysis, to facilitate the identification of the nature of backscattering atoms, and to conventional EXAFS data fitting. The collective results showed the existence of two pools of iron: mononuclear Fe(III)-NOM complexes and precipitated Fe(III) (hydr)oxides. In the soil with lowest pH (4.6) and Fe content mononuclear organic complexes were the completely dominating fraction whereas in soils with higher pH and Fe content increasing amounts of Fe (hydr)oxides were detected. These results are of environmental importance, as the different iron pools most likely have markedly different reactivities.     

Use of the selective agar medium CREAD for monitoring the level of airborne spoilage moulds in cheese production

It was investigated if a selective medium for common cheese spoiling moulds (CREAD) could give more relevant information than a general mould medium in hygienic air-sampling in cheese factories. A total of 126 air-samples were taken in six Nordic cheese factories using the general mould medium DG18 and CREAD. The level and genera of air-borne mould was determined. Identification to species-level was performed for a selection of samples. In five cheese factories the mycobiota was dominated by Penicillium spp. and in one cheese factory by Cladosporium spp. The concentration of air-borne moulds varied between the cheese factories ranging from 1 to 270 cfu/m3 on DG18 with a median value of 17. The number of mould colonies was in general lower at CREAD. Identification indicated that CREAD supported growth of common spoilage moulds for cheese, such as Penicillium palitans and P. commune. The mycobiota on DG18 also consisted of moulds not commonly associated with spoilage of cheese, such as Cladosporium spp., P. brevicompactum and P. chrysogenum. Contamination of cheese with mould is periodically a problem in production of semi-hard cheese and the level of air-borne mould is therefore routinely monitored in cheese factories. A clear correlation between the total number of moulds in air and mould growth on products is not always found. The conclusion from the investigation is that it is recommended to use a selective medium for cheese spoilage moulds, such as CREAD in hygienic monitoring.     

Genetic parameters of forage dry matter intake and milk produced from forage in Swedish Red and Holstein dairy cows

High-yielding dairy cows are often fed high proportions of cereal grain and pulses. For several reasons, it would be desirable to replace these feed sources with forage, which is not suitable for human consumption. Feeding large amounts of forage to dairy cows could also make dairy production more publicly acceptable in the future. In this study, we estimated genetic parameters for total dry matter intake (DMI), DMI from forage (DMI_For), energy-corrected milk (ECM), and ECM produced from forage (ECM_For). A total of 1,177 lactations from 575 cows of Swedish Red (SR) and Holstein (HOL) dairy breeds were included in the study. Mixed linear animal random regression models were used, with fixed effect of calving season and lactation week nested within parity 1 and 2+, fixed effect of calving year, and random regression coefficients for breeding value (up to linear) and permanent environmental effect (up to quadratic) of the cow. Heritability for DMI and DMI_For was generally higher for HOL than for SR in all-parity data and in later parities; however, the opposite was true for first parity. Heritability for DMI and DMI_For during the first 8 wk averaged 0.11 and 0.15, respectively, in all-parity data for the 2 breeds. Corresponding values for ECM_For and ECM were 0.21 and 0.29, respectively. In first parity, values were 0.32, 0.36, 0.28, and 0.51, respectively. The genetic correlation between DMI and DMI_For was high, above 0.83, and fairly constant across the lactation. The genetic correlation between ECM_For and ECM was close to unity in the later part of lactation for both breeds, but was around 0.8 in the early lactation for both breeds; it decreased for HOL to 0.54 in wk 17. The genetic correlations between DMI and ECM_For and between DMI_For and ECM_For were low and negative for HOL (absolute value ～0.2–0.3), but changed for SR from weakly positive in early lactation to negative values and back to positive toward the end of lactation. For most traits, the correlation between wk 1 and wk 8 into the lactation was very high; the lowest value was for DMI in HOL at 0.81. The genetic correlation between parities was rather high in the first part of the lactation. During the first 8 wk, the correlation was lower for HOL than for SR, except for ECM. We found that DMI_For and ECM_For showed reasonably large heritability, and future work should explore the possibility of genomic evaluations.     

Topographic quantification of non-planar localized surfaces

The topography of non-planar localized surfaces can be quantified by roughness indices as well as by height and angular distributions of the surface elements. For the roughness indices general relationships between the profile and spatial quantities including overlapping have been established. Some measurement methods are briefly considered. The profile analysis has proved to be a reliable, easy and fast way to characterize the topography of surfaces.