Isolation of adipophilin and butyrophilin from bovine milk and characterization of a cDNA encoding adipophilin

The milk fat globule membrane-associated proteins adipophilin (alias adipocyte differentiation-related protein) and butyrophilin were purified from bovine milk by reverse-phase chromatography. The nucleotide sequence of bovine adipophilin was obtained via peptide mapping and sequencing of a mammary gland cDNA clone, which comprises 1841 nucleotides and has an open reading frame of 450 amino acids. By peptide mapping, 19% of the amino acid sequence was confirmed. The obtained amino acid sequence has 87 and 80% identical residues with human and mouse adipophilin, respectively. Alignment with the proteins perilipin and TIP47 revealed two highly conserved segments, which may assemble into amphipathic alpha-helices.     

Active packaging of cheese with allyl isothiocyanate, an alternative to modified atmosphere packaging

The natural antimicrobial compound allyl isothiocyanate (AITC), found in mustard oil, is effective against cheese-related fungi both on laboratory media and cheese. Penicillium commune, Penicillium roqueforti, and Aspergillus flavus were more sensitive to AITC when it was added just after the spores had completed 100% germination and branching had started on Czapek yeast extract agar than were spores in the dormant phase. The use of 1 AITC label (Wasaouro interior labels, LD30D, 20 by 20 mm) in combination with atmospheric air in the packaging extended the shelf life of Danish Danbo cheese from 4 1/2 to 13 weeks. Two AITC labels extended the shelf life from 4 1/2 to 28 weeks. Both 1 and 2 labels in combination with modified atmosphere packaging extended the shelf life of the cheese from 18 to 28 weeks. This study showed that AITC was absorbed in the cheese, but it was not possible to detect any volatile breakdown products from AITC in the cheese. Cheese stored for up to 12 weeks with an AITC label had an unacceptable mustard flavor. The mustard flavor decreased to an acceptable level between weeks 12 and 28. Cheese stored in atmospheric air had a fresher taste without a CO2 off-flavor than did cheese stored in modified atmosphere packaging. AITC may be a good alternative to modified atmosphere packaging for cheese. The extended shelf life of cheese in the package is very desirable: the cheese can be transported longer distances, and the packaging can be used for the final maturing of the cheese. Furthermore, AITC can address problems such as pinholes and leaking seals in cheese packaging.     

Nonstationary Cointegration in the Fractionally Cointegrated VAR Model

We consider the fractional cointegrated vector autoregressive (CVAR) model of Johansen and Nielsen (2012a) and make two distinct contributions. First, in their consistency proof, Johansen and Nielsen (2012a) imposed moment conditions on the errors that depend on the parameter space, such that when the parameter space is larger, stronger moment conditions are required. We show that these moment conditions can be relaxed, and for consistency we require just eight moments regardless of the parameter space. Second, Johansen and Nielsen (2012a) assumed that the cointegrating vectors are stationary, and we extend the analysis to include the possibility that the cointegrating vectors are non‐stationary. Both contributions require new analysis and results for the asymptotic properties of the likelihood function of the fractional CVAR model, which we provide. Finally, our analysis follows recent research and applies a parameter space large enough that the usual (non‐fractional) CVAR model constitutes an interior point and hence can be tested against the fractional model using a Chi‐squared‐test.     

Stochastic response of hysteretic systems

This paper deals with approximate stochastic response of hyseretic structures under white noise excitation, based on It stochastic differential equations. Instead of the original system an equivalent nonlinear system is considered, in which the drift vector is given by a series expansion of the order n 1 in terms of the state variables. n = 1 represents the well-known case of equivalent linearization. Components in the drift vector representing the nonanalytical constitutive equations are replaced by a cubic polynomial expansion. The hierarchy of statistical moment equations is closed by a cumulant neglect closure scheme. The method has been applied to a bilinear single degree-of-freedom system subjected to white noise excitation, for which an equivalent system with a cubic series expansion to the constitutive equation is considered. The results obtained are compared with those of numerical simulation and alternative methods, and they provide substantial improvements compared to equivalent linearization.     

Characterization of activated sludge flocs by confocal laser scanning microscopy and image analysis

In this study we present a new approach to determine volumes, heterogeneity factors, and compositions of the bacterial population of activated sludge flocs by 3D confocal imaging. After staining the fresh flocs with fluorescein-isothiocyanate, 75 stacks of images (containing approx. 3000 flocs) were acquired with a confocal laser scanning microscope. The self-developed macro 3D volume and surface determination for the KS 400 software package combined the images of one stack to a 3D image and calculated the real floc volume and surface. We determined heterogeneity factors like the ratio of real floc surface to the surface of a sphere with the respective volume and the fractal dimension (D(f)). According to their significant influence on floc integrity and quality, we also investigated the chemical composition of flocs and quantified their bacterial population structure by using group-specific rRNA-targeted probes for fluorescence in situ hybridization. By a settling experiment we enriched flocs with poor settling properties and determined the above-mentioned parameters. This approach revealed shifts in floc volume, heterogeneity, and bacterial and chemical composition according to the settling quality of the flocs.     

A digital system for surface roughness analysis of plane and cylindrical parts

This paper describes an on-line system for digital analysis of surfaces where the electrical signal from a surface roughness instrument is converted and input into a desk-top computer which also controls the traversing. Special fixtures integrated with the system and dedicated software permit two- and three-dimensional tracings to be carried out on plane as well as on cylindrical parts. The surface profiles can be plotted in different ways. Roughness parameters, frequency spectra etc, can be computed digitally and all results can be stored. Scratches and irregularities on the surface can be identified and processed separately from the rest of the surface. The desk-top computer is connected to the university mainframe computer through a modem, this being of advantage in connection with graphic data processing. The capability of the equipment has been investigated and results from calibration are presented.     

A New Immobilized Glucose Isomerase with High Productivity Produced by a Strain of Streptomyces murinus

The properties of a new immobilized glucose isomerase, produced by a selected strain of Streptomyces murinus, are described. The major advantages of the new immobilized enzyme are a productivity of more than 10,000 kg syrup dry substance per kg enzyme under optimal industrial conditions, increased activity and a very low syrup by-product formation. The influence of process parameters (temperature, pH and feed syrup additives) on activity and stability is discussed based on laboratory and industrial plant data.     

Liquefaction,Saccharification and Isomerization of Starches from Sources Other than Maize

Examination of the suitability of a number of starches for the production of fructose syrup is described. In addition to maize starch, starch from wheat, potatoes and tapioca has been examined. Liquefaction, saccharification and purification were performed in pilot scale, whereas the produced syrups were isomerized in laboratory scale. The liquefactions were performed with Termamyl® in a jet cooker process. Saccharification with Amyloglucosidase Novo 150 L under standard conditions gave good DX values. After purification the syrups were isomerized in laboratory columns with Sweetzyme®. The results - activity and productivity of Sweetzyme - were for all syrups as good as or better than those obtained with dextrose solutions.     

A longitudinal observational study of the dynamics of Mycoplasma bovis antibodies in naturally exposed and diseased dairy cows

Mycoplasma bovis is an important pathogen causing disease and substantial economic losses in cattle. However, knowledge of the dynamics of antibody responses in individual cows in the face of an outbreak is currently extremely limited. The use of commercial antibody tests to support clinical decision-making and for surveillance purposes is therefore challenging. Our objective was to describe the dynamics of M. bovis antibody responses in 4 Danish dairy herds experiencing an acute outbreak of M. bovis-associated disease, and to compare the antibody dynamics between dairy cows with different disease manifestations. A total of 120 cows were examined using a standardized clinical protocol and categorized into 4 disease groups: “mastitis,” “systemic,” “nonspecific,” and “none.” Paired blood and milk samples were collected and tested using a commercial M. bovis antibody–detecting ELISA. Plots of raw data and generalized additive mixed models with cow and herd as random effects were used to describe serum and milk antibody dynamics relative to the estimated time of onset of clinical disease. Cows with mastitis had high optical density measurement (ODC%) of antibodies in both milk and serum at disease onset. The estimated mean ODC% in milk was below the manufacturer's cut-off for the other groups for the entire study period. The estimated mean serum ODC% in the “systemic” group was high at onset of disease and stayed above the cut-off until 65 d after disease onset. However, the lower 95% confidence interval (CI) for the mean ODC% was only above the manufacturer's cut-off between 7 and 17 d after onset of disease. The CI of the “systemic” and “none” groups did not overlap at any time between the day of disease onset and 65 d after disease onset, and the estimated mean ODC% for both the “nonspecific” and “none” groups were generally below the cut-off for the majority of the study period. In conclusion, the serum antibody responses were highly dynamic and showed a high level of variation between individual cows. This strongly suggests that serology is unlikely to be useful for individual diagnosis of M. bovis-associated disease in dairy cows. However, it might still be useful for herd- or group-level diagnosis. Antibodies in milk were only increased in cows with M. bovis mastitis, indicating that milk antibody measurements only have diagnostic utility for cows with mastitis.