Ulf Andersen

Protein denaturation of whey protein isolates (WPIs) induced by high intensity ultrasound during heat gelation

In this study, the impact of high intensity ultrasound (HIU) on proteins in whey protein isolates was examined. Effects on thermal behavior, secondary structure and nature of intra- and intermolecular bonds during heat-induced gelling were investigated. Ultrasonication (24 kHz, 300 W/cm(2), 2078 J/mL) significantly reduced denaturation enthalpies, whereas no change in secondary structure was detected by circular dichroism. The thiol-blocking agent N-ethylmaleimide was applied in order to inhibit formation of disulfide bonds during gel formation. Results showed that increased contents of α-lactalbumin (α-La) were associated with increased sensitivity to ultrasonication. The α-La:β-lactoglobulin (β-Lg) ratio greatly affected the nature of the interactions formed during gelation, where higher amounts of α-La lead to a gel more dependent on disulfide bonds. These results contribute to clarifying the mechanisms mediating the effects of HIU on whey proteins on the molecular level, thus moving further toward implementing HIU in the processing chain in the food industry.

Effect of Gel Firmness at Cutting Time, pH, and Temperature on Rennet Coagulation and Syneresis: An in situ 1H NMR Relaxation Study

The objective of this study was to monitor rennet-induced milk gel formation and mechanically induced gel syneresis in situ by low-field NMR. pH, temperature, and gel firmness at cutting time were varied in a factorial design. The new curve-fitting method Doubleslicing revealed that during coagulation two proton populations with distinct transverse relaxation times (T2,1=181, T2,2=465 ms) were present in fractions (f1=98.9%, f2=1.1%). Mechanical cutting of the gel in the NMR tube induced macrosyneresis, which led to the appearance of an additional proton population (T2,3=1500-2200 ms) identified as whey. On the basis of NMR quantification of whey water the syneresis rate was calculated and found to be significantly dependent on pH and temperature.

Water Properties in Cream Cheeses with Variations in pH, Fat, and Salt Content and Correlation to Microbial Survival

Water mobility and distribution in cream cheeses with variations in fat (4, 15, and 26%), added salt (0, 0.625, and 1.25%), and pH (4.2, 4.7, and 5.2) were studied using (1)H NMR relaxometry. The cheese samples were inoculated with a mixture of Listeria innocua, Escherichia coli O157 and Staphylococcus aureus, and partial least-squares regression revealed that (1)H T(2) relaxation decay data were able to explain a large part of the variation in the survival of E. coli O157 (64-83%). However, the predictions of L. innocua and S. aureus survival were strongly dependent on the fat/water content of the samples. Consequently, the present results indicate that NMR relaxometry is a promising technique for predicting the survival of these bacteria; however, the characteristics of the sample matrix are substantial.

Physical sample structure as predictive factor in growth modeling of Listeria innocua in a white cheese model system

Growth of Listeria innocua at 9 °C was investigated in white cheeses manufactured from ultra-filtrate milk concentrate added varying amounts of skimmed milk powder, NaCl and glucono-delta-lactone. Characterization of the white cheese structures was performed using nuclear magnetic resonance (NMR) T₂ relaxation parameters (relaxation times constants, relative areas and width of peaks) and their applicability as predictive factors for maximum specific growth rate, √μ(max) and log-increase in 6 weeks of L. innocua was evaluated by polynomial modeling. Inclusion of NMR parameters was able to increase the goodness-of-fit of two basic models; one having pH, undissociated gluconic acid (GA(u), mM) and NaCl (% w/v) as predictive factors and another having pH, GA(u) and a(w) as predictive factors. However, the best model fit was observed using √μ(max) as response for the model including pH, GA(u), aw and Width T₂₁ revealing the lowest relative root mean squared errors of 14.0%. As the T₂ relaxation population T₂₁ is assigned to represent immobilized bulk water protons and the width T₂₁ the heterogeneity of this water population, growth of L. innocua in white cheese seemed to be dependent on the heterogeneity of the immobilized bulk water present in cheese.

Microstructure and Material Properties of Milk Fat Systems During Temperature Fluctuations

Microstructure and material properties of milk fat systems containing different amount of milk fat globules and different emulsion types were studied towards the effects of temperature fluctuations stress. The three systems were (i) an emulsion based on anhydrous milk fat (AMF) consisting of crystals dispersed in liquid oil, (ii) butter which is a water in oil emulsion consisting of a continuous crystal network with partially disrupted fat globules interrupted by intact fat globules and water droplets, and (iii) freeze-dried cream which is an oil in water emulsion consisting of concentrated milk fat globules. The microstructure of the systems was observed by confocal laser scanning microscopy and linked to their material properties by large and small deformation rheology techniques. Different rheological techniques were used to characterize the material descriptors. The crystallization behavior was studied by thermal analysis and pulsed nuclear magnetic resonance. The freeze-dried cream was characterized as the softest, more elastic, and less stiff when compared to butter and AMF. Temperature fluctuations led to drastic material consolidation in butter as a consequence of flocculation of fat globules and formation of a more dense crystal network with stronger bonds. Only after several temperature fluctuations, material consolidation was observed in AMF, whereas no significant changes in material properties were observed in the freeze-dried cream. The intact fat globules decrease the hardness and stiffness, whereas the amount and type of contact points between crystal clusters of the dispersed crystal phase contribute to increase hardness, stiffness, and brittleness of the systems.

Molecular Water Motions of Skim Milk Powder Solutions during Acidification Studied by 17O and 1H Nuclear Magnetic Resonance and Rheology

The molecular motion of water was studied in glucono-δ-lactone-acidified skim milk powder (SMP) solutions with various pH values and dry matter contents. NMR relaxometry measurements revealed that lowering the pH in SMP solutions affected 17O and 1H T2 relaxation rates almost identically. Consequently, the present study indicates that the proteins present in the samples do not affect the 1H relaxation behavior markedly, even at relatively high SMP concentrations (15-25%). Comparison of rheological measurements and NMR measurements suggested that the collapse of κ-casein during acidification could contribute to the initial decrease in 17O and 1H relaxation rate in the pH range between 6.6 and 5.5 for 15% SMP and in the pH range between 6.6 and 5.9 for 25% SMP. However, below pH 5.5 the viscosity and 17O and 1H NMR relaxation rates did not correlate, revealing that the aggregation of casein micelles, which increases viscosity below pH 5.5, does not involve major repartitioning of water.

Impact of NaCl reduction in Danish semi-hard Samsoe cheeses on proliferation and autolysis of DL-starter cultures

Reduction of sodium chloride (NaCl) in cheese manufacturing is a challenge for the dairy industry. NaCl has a profound role on microbial development influencing cheese sensory and technological properties. The purpose of this work was to investigate how proliferation, distribution and autolysis of two commercial DL-starter cultures (C1 and C2) used in the production of Danish semi-hard Samsoe cheeses were affected by reduced NaCl levels. Cheeses containing <0.3% (unsalted), 2.3% (reduced-salt) and 3.4% (normal-salted) (w/v) NaCl in moisture were produced and analyzed during 12 weeks of ripening. Lactic acid bacteria (LAB), distribution of bacteria as single cells or microcolonies, their viability in the cheeses and cell autolysis were monitored during ripening, as well as the impact of NaCl content and autolysis on the formation of free amino acids (FAA). Reduction of NaCl resulted in higher LAB counts at the early stages of ripening, with differences between the two DL-starter cultures. The unsalted cheeses produced with C1 had retained a significantly higher number of the initial LAB counts (cfu/g) after 1 and 2 weeks of ripening (i.e. 58% and 71%), compared to the normal-salted cheeses (i.e. 22% and 21%), whereas no significant difference was found between the reduced-salt (i.e. 31% and 35%) and normal-salted cheeses. At the later stages of ripening (i.e. 7 and 11 weeks) NaCl had no significant influence. For cheeses produced with C2, a significant influence of NaCl was only found in cheeses ripened for 7 weeks, where the unsalted and reduced-salt cheeses had retained a significantly higher number of the initial LAB counts (cfu/g) (i.e. 39% and 38%), compared to the normal-salted cheeses (i.e. 21%). In the Samsoe cheeses, bacteria were organized as single cells, in groups of 2-3 cells or in groups of ≥4 cells. During ripening the decrease in the number of viable bacteria was mainly due to a reduction in the number of viable bacteria organized in groups of ≥4 cells. A negative correlation between NaCl content and PepX activity was observed. At the end of ripening the total FAA content was lower in the unsalted cheeses, compared to the reduced- and normal-salted cheeses. In conclusion, NaCl had a significant influence on proliferation of both DL-starter cultures. However, the influence of NaCl on culture development was more pronounced in cheeses produced with DL-starter culture C1. As both texture and taste are parameters known to be affected by the development of the starter culture, the design of starter cultures for reduced NaCl cheeses is recommended.

The effect of butter grains on physical properties of butter-like emulsions

Milk fat exists as globules in its natural state in milk. The potential of using globular fat to modulate the rheological properties and crystallization behavior in butter-like emulsions was studied in the present work. We conducted a comparative study of butter-like emulsions, with a fat phase consisting of 0, 10, 25, 50, or 100% anhydrous milk fat (AMF), the remaining fat being butter grains, and all samples containing 20% water, to obtain systematic variation in the ratio of globular fat. All emulsions were studied over 4wk of storage at 5°C. By combining small and large deformation rheology, we conducted a detailed characterization of the rheological behavior of butter-like emulsions. We applied differential scanning calorimetry to monitor thermal behavior, confocal laser scanning microscopy for microstructural analysis, and low-field pulsed nuclear magnetic resonance spectrometry to measure solid fat content. By combining these techniques, we determined that increasing the fraction of globular fat (by mixing with butter grains) decreases the hardness of butter-like emulsions up to an order of magnitude at d 1. However, no difference was observed in thermal behavior as a function of butter grain content, as all emulsions containing butter grains revealed 2 endothermal peaks corresponding to the high (32.7°C ± 0.6) and medium (14.6°C ± 0.1) melting fractions of fatty acids. In terms of microstructure, decreasing the amount of butter grains in the emulsions resulted in formation of a denser fat crystal network, corresponding to increased hardness. Moreover, microstructural analysis revealed that the presence of butter grains resulted in faster formation of a continuous fat crystal network compared with the 100% AMF sample, which was dominated by crystal clusters surrounded by liquid oil. During storage, hardness remained stable and no changes in thermal behavior were observed, despite an increase in solid fat content of up to 5%. After 28d of storage, we observed no difference in either microstructural or rheological properties, indicating that formation of primary bonds occurs primarily within the first day of storage. The rheological behavior of butter-like emulsions is not determined solely by hardness, but also by stiffness related to secondary bonds within the fat crystal network. The complex rheological behavior of milk fat-based emulsions is better characterized using multiple parameters.

Evaluation of Yogurt Microstructure Using Confocal Laser Scanning Microscopy and Image Analysis

The microstructure of protein networks in yogurts defines important physical properties of the yogurt and hereby partly its quality. Imaging this protein network using confocal scanning laser microscopy (CSLM) has shown good results, and CSLM has become a standard measuring technique for fermented dairy products. When studying such networks, hundreds of images can be obtained, and here image analysis methods are essential for using the images in statistical analysis. Previously, methods including gray level co-occurrence matrix analysis and fractal analysis have been used with success. However, a range of other image texture characterization methods exists. These methods describe an image by a frequency distribution of predefined image features (denoted textons). Our contribution is an investigation of the choice of image analysis methods by performing a comparative study of 7 major approaches to image texture description. Here, CSLM images from a yogurt fermentation study are investigated, where production factors including fat content, protein content, heat treatment, and incubation temperature are varied. The descriptors are evaluated through nearest neighbor classification, variance analysis, and cluster analysis. Our investigation suggests that the texton-based descriptors provide a fuller description of the images compared to gray-level co-occurrence matrix descriptors and fractal analysis, while still being as applicable and in some cases as easy to tune.

Thyroid Autoantibodies in Thyroid Cancer

It has previously been shown that thyroid autoantibodies are frequent in patients treated for differentiated thyroid cancer 1,2 Whether these antibodies are due to therapy or are involved in the pathogenesis of the disease has, however, not yet been clarified. We were therefore interested in studying these antibodies in thyroid cancer before and after treatment.