Helena Lindmark-Månsson

Antioxidative factors in milk

Lipid auto-oxidation in milk is affected by a complex interplay of pro- and antioxidants. Several of these compounds are also important nutrients in the human diet and may have other physiological effects in the gastrointestinal tract and other tissues. Among antioxidative enzymes superoxide dismutase catalyses the dismutation of superoxide anion to hydrogen peroxide. The degradation of hydrogen peroxide can be catalysed by catalase and the selenoprotein glutathione peroxidase. The latter enzyme can also degrade lipid peroxides. Lactoferrin may have an important role by binding pro-oxidative iron ions. The occurrence of different forms of these antioxidative proteins in milk and available data on their functional role are reviewed. More remains to be learnt of individual compounds and as an example the potential role of seleno compounds in milk is virtually unknown. Antioxidative vitamins in milk can provide an important contribution to the daily dietary intake. Moreover vitamin E and carotenoids act as fat-soluble antioxidants, e.g. in the milk fat globule membrane, which is regarded as a major site of auto-oxidation. Vitamin C is an important water-soluble antioxidant and interacts in a complex manner with iron and fat-soluble antioxidants. The concentrations of these compounds in milk are affected by cow feeding rations and milk storage conditions. Since milk contains a number of antioxidants many reactions are possible and the specific function of each antioxidant cannot easily be defined. There are indications that other compounds may have antioxidative function and measurement of total antioxidative capacity should be a useful tool in evaluating their relative roles.

The Role of Osteopontin in Tumor Progression and Metastasis in Breast Cancer

The use of cancer biomarkers to anticipate the outlines of disease has been an emerging issue, especially as cancer treatment has made such positive steps in the last few years. Progress in the development of consistent malignancy markers is imminent because advances in genomics and bioinformatics have allowed the examination of immense amounts of data. Osteopontin is a phosphorylated glycoprotein secreted by activated macrophages, leukocytes, and activated T lymphocytes, and is present in extracellular fluids, at sites of inflammation, and in the extracellular matrix of mineralized tissues. Several physiologic roles have been attributed to osteopontin, i.e., in inflammation and immune function, in mineralized tissues, in vascular tissue, and in kidney. Osteopontin interacts with a variety of cell surface receptors, including several integrins and CD44. Binding of osteopontin to these cell surface receptors stimulates cell adhesion, migration, and specific signaling functions. Overexpression of osteopontin has been found in a variety of cancers, including breast cancer, lung cancer, colorectal cancer, stomach cancer, ovarian cancer, and melanoma. Moreover, osteopontin is present in elevated levels in the blood and plasma of some patients with metastatic cancers. Therefore, suppression of the action of osteopontin may confer significant therapeutic activity, and several strategies for bringing about this suppression have been identified. This review looks at the recent advances in understanding the possible mechanisms by which osteopontin may contribute functionally to malignancy, particularly in breast cancer. Furthermore, the measurement of osteopontin in the blood or tumors of patients with cancer, as a way of providing valuable prognostic information, will be discussed based on emerging clinical data. (Cancer Epidemiol Biomarkers Prev 2007;16(6):1087–97)

Immunoglobulins, growth factors and growth hormone in bovine colostrum and the effects of processing

In colostrum collected 0-80 h postpartum the contents of immunoglobulins (Igs), transforming growth factor beta-2 (TGF-beta2), insulin-like growth factor-1 (IGF-1) and growth hormone (GH) were analysed. Colostrum initially contained 90 mg mL(-1) IgG1, 2.8 mg mL(-1) IgG2, 1.6 mg mL(-1) IgA, 4.5 mg mL(-1) IgM, and these concentrations declined by 92%, 87%, 93% and 84%, respectively, in the samples collected later. Of the growth factors, colostrum initially contained 289-310 ng mL(-1) TGF-beta2 and the concentration diminished to 66 ng mL(-1). The content of IGF-1 and GH postpartum decreased from 870 to 150 ng mL(-1), and from 0.17 to < 0.03 ng mL(-1), respectively. Heat treatment and freeze-drying of colostral whey decreased the content of Igs to 75%, while the contents of IGF-1 and TGF-beta2 were unaffected. A similar processing, including filtration steps reduced also the IGF-1 and TGF-beta2 by 25%. IgM seems to be the most sensitive of the Igs to processing

Antioxidant capacity of bovine milk as assayed by spectrophotometric and amperometric methods

The total antioxidant capacity (TAC) of bovine milk, whey, and low-molecular-weight (LMW) fractions of whey was investigated using spectrophotometric methods including an ABTS-based method (reduction of the cation radical of 2,2′-azinobis(3-ethylenebenzothiazoline-6-sulfonic acid)) and a FRAP method (reduction of Fe3+). Significant antioxidant capacity in milk and whey was demonstrated by the ABTS method (TAC(ABTS)), and it apparently increased with increasing pH. TAC (ABTS) was several-fold higher in milk than in whey, which had a slightly higher TAC than a LMW fraction prepared from it. Also the FRAP method could be used to demonstrate TAC in whey, although the low pH necessary for this method led to some protein precipitation. Most of the ferric-reducing activity of whey was found in the LMW fraction. The TAC values obtained using these methods were also compared to those obtained using a newly developed flow-injection amperometric (FIAmp) procedure for LMW fractions. High correlations were found for the TAC values of LMW samples obtained by the ABTS, FRAP and FIAmp methods (R2>0.8, P<0.001). Furthermore, to identify the major antioxidants in the LMW fraction, it was treated by uricase. Since most of the TAC (using ABTS, FRAP and FIAmp methods) was removed by uricase treatment, it could be concluded that urate is the major antioxidant in the LMW fraction. Also the effect of heat treatment on TAC in milk and whey was monitored. After heating of whey at 63°C for 1 h, TAC(ABTS) tended to increase by at least 20%, while TAC(FRAP) was not significantly changed. In milk, TAC(ABTS) remained constant during the same heat treatment. It is concluded that the methods tested gave the most reliable results for the LMW fraction of whey and that the use of several methods is necessary to characterise the antioxidant capacity of milk.

Nutrient density of beverages in relation to climate impact.

The food chain contributes to a substantial part of greenhouse gas (GHG) emissions and growing evidence points to the urgent need to reduce GHGs emissions worldwide. Among suggestions were proposals to alter food consumption patterns by replacing animal foods with more plant-based foods. However, the nutritional dimensions of changing consumption patterns to lower GHG emissions still remains relatively unexplored. This study is the first to estimate the composite nutrient density, expressed as percentage of Nordic Nutrition Recommendations (NNR) for 21 essential nutrients, in relation to cost in GHG emissions of the production from a life cycle perspective, expressed in grams of CO2-equivalents, using an index called the Nutrient Density to Climate Impact (NDCI) index. The NDCI index was calculated for milk, soft drink, orange juice, beer, wine, bottled carbonated water, soy drink, and oat drink. Due to low-nutrient density, the NDCI index was 0 for carbonated water, soft drink, and beer and below 0.1 for red wine and oat drink. The NDCI index was similar for orange juice (0.28) and soy drink (0.25). Due to a very high-nutrient density, the NDCI index for milk was substantially higher (0.54) than for the other beverages. Future discussion on how changes in food consumption patterns might help avert climate change need to take both GHG emission and nutrient density of foods and beverages into account.

Potential role of milk fat globule membrane in modulating plasma lipoproteins, gene expression, and cholesterol metabolism in humans: a randomized study

BACKGROUND Butter is rich in saturated fat [saturated fatty acids (SFAs)] and can increase plasma low density lipoprotein (LDL) cholesterol, which is a major risk factor for cardiovascular disease. However, compared with other dairy foods, butter is low in milk fat globule membrane (MFGM) content, which encloses the fat. We hypothesized that different dairy foods may have distinct effects on plasma lipids because of a varying content of MFGM. OBJECTIVE We aimed to investigate whether the effects of milk fat on plasma lipids and cardiometabolic risk markers are modulated by the MFGM content. DESIGN The study was an 8-wk, single-blind, randomized, controlled isocaloric trial with 2 parallel groups including overweight men and women (n = 57 randomly assigned). For the intervention, subjects consumed 40 g milk fat/d as either whipping cream (MFGM diet) or butter oil (control diet). Intervention foods were matched for total fat, protein, carbohydrates, and calcium. Subjects were discouraged from consuming any other dairy products during the study. Plasma markers of cholesterol absorption and hepatic cholesterol metabolism were assessed together with global gene-expression analyses in peripheral blood mononuclear cells. RESULTS As expected, the control diet increased plasma lipids, whereas the MFGM diet did not [total cholesterol (±SD): +0.30 ± 0.49 compared with -0.04 ± 0.49 mmol/L, respectively (P = 0.024); LDL cholesterol: +0.36 ± 0.50 compared with +0.04 ± 0.36 mmol/L, respectively (P = 0.024); apolipoprotein B:apolipoprotein A-I ratio: +0.03 ± 0.09 compared with -0.05 ± 0.10 mmol/L, respectively (P = 0.007); and non-HDL cholesterol: +0.24 ± 0.49 compared with -0.14 ± 0.51 mmol/L, respectively (P = 0.013)]. HDL-cholesterol, triglyceride, sitosterol, lathosterol, campesterol, and proprotein convertase subtilisin/kexin type 9 plasma concentrations and fatty acid compositions did not differ between groups. Nineteen genes were differentially regulated between groups, and these genes were mostly correlated with lipid changes. CONCLUSIONS In contrast to milk fat without MFGM, milk fat enclosed by MFGM does not impair the lipoprotein profile. The mechanism is not clear although suppressed gene expression by MFGM correlated inversely with plasma lipids. The food matrix should be considered when evaluating cardiovascular aspects of different dairy foods. This trial was registered at clinicaltrials.gov as NCT01767077.

Lactoferricin treatment decreases the rate of cell proliferation of a human colon cancer cell line

Food components modify the risk of cancer at a large number of sites but the mechanism of action is unknown. In the present investigation, we studied the effect of the peptide lactoferricin derived from bovine milk lactoferrin on human colon cancer CaCo-2 cells. The cells were either untreated or treated with 2.0, 0.2, or 0.02 microM lactoferricin. Cell cycle kinetics were investigated with a bromodeoxyuridine DNA flow cytometric method. The results show that lactoferricin treatment slightly but significantly prolonged the S phase of the cell cycle. Lactoferricin treatment lowered the level of cyclin E1, a protein involved in the regulation of genes required for G(1)/S transition and consequently for efficient S phase progression. The slight prolongation of the S phase resulted in a reduction of cell proliferation, which became more apparent after a long treatment time.

Bovine chromosomal regions affecting rheological traits in acid-induced skim milk gels

The production of fermented milk products has increased worldwide during the last decade and is expected to continue to increase during the coming decade. The quality of these products may be optimized through breeding practices; however, the relations between cow genetics and technological properties of acid milk gels are not fully known. Therefore, the aim of this study was to identify chromosomal regions affecting acid-induced coagulation properties and possible candidate genes. Skim milk samples from 377 Swedish Red cows were rheologically analyzed for acid-induced coagulation properties using low-amplitude oscillation measurements. The resulting traits, including gel strength, coagulation time, and yield stress, were used to conduct a genome-wide association study. Single nucleotide polymorphisms (SNP) were identified using the BovineHD SNPChip (Illumina Inc., San Diego, CA), resulting in almost 621,000 segregating markers. The genome was scanned for putative quantitative trait loci (QTL) regions, haplotypes based on highly associated SNP were inferred, and the additive genetic effects of haplotypes within each QTL region were analyzed using mixed models. A total of 8 genomic regions were identified, with large effects of the significant haplotype explaining between 4.8 and 9.8% of the phenotypic variance of the studied traits. One major QTL was identified to overlap between gel strength and yield stress, the QTL identified with the most significant SNP closest to the gene coding for κ-casein (CSN3). In addition, a chromosome-wide significant region affecting yield stress on BTA 11 was identified to be colocated with PAEP, coding for β-lactoglobulin. Furthermore, the coagulation properties of the genetic variants within the 2 genes were compared with the coagulation properties identified by the patterns of the haplotypes within the regions, and it was discovered that the haplotypes were more diverse and in one case slightly better at explaining the phenotypic variance. Besides these significant QTL comprising the 2 milk proteins, 3 additional genes are proposed as possible candidates, namely RAB22A, CDH13, and STAT1, and all have previously been found to be expressed in the mammary gland. To our knowledge, this is the first attempt to map QTL regions for acid-induced coagulation properties.

Significance of Osteopontin Expression in Human Invasive Breast Tumour Stroma

Osteopontin has been reported to stimulate cell adhesion, migration and specific signalling functions. Its over- expression has been found in melanoma, breast, lung, colorectal, stomach and ovarian cancer. However, its overexpres- sion and role in human breast cancer remains to be elucidated. In this study, invasive breast tumours from 129 patients were examined by immunohistochemistry in order to assess osteopontin association with several molecular tumour mark- ers. Additionally, its relationship with proliferation and angiogenesis was determined. Ultimately, other tumour variables such as histological grade, tumour size and nodal status were also assessed. Results achieved showed that no statistical significant association exists between osteopontin expression and major clinicopathological parameters or angiogenesis, except for the number of lymph nodes involved. However, a correlation with some molecular markers was observed, namely with P-Cadherin, EGFR, cytokeratin 14 and vimentin. Additionally, higher proliferation rates were found for the tumours expressing osteopontin. Although several studies refer osteopontin as a potential breast cancer biomarker, it is still not clear if it can provide important diagnosis information, evaluate treatment effects or assess the potential for metas- tatic disease in patients.

Reduction of ultraviolet light-induced DNA damage in human colon cancer cells treated with a lactoferrin-derived peptide

Treatment of Caco-2 cells with the peptide lactoferricin(4-14), results in reduction of the growth rate by prolongation of the S phase of the cell cycle. Lactoferricin(1-25) is formed in the gut by cleavage from lactoferrin and the bioactive amino acids are found within lactoferricin(4-14). Our hypothesis is that the reduction of the rate of S phase progression may result in increased DNA repair. To test this hypothesis, Caco-2 cells were subjected to UV light that caused DNA lesions and then the cells were grown in the absence or presence of 2.0 μM lactoferricin(4-14). Evaluation of DNA strand breaks using the comet assay showed that lactoferricin(4-14) treatment indeed resulted in a reduction of comets showing damaged DNA. In the search for a mechanism, we have investigated the levels of several proteins involved in cell cycle regulation, DNA replication, and apoptosis using Western blot. Lactoferricin(4-14) treatment resulted in an increased expression of flap endonuclease-1 pointing to increased DNA synthesis activity. Lactoferricin(4-14) treatment decreased the expression of the proapoptotic protein B-cell lymphoma 2-associated X protein (or Bax), indicating decreased cell death. As we have found previously, lactoferricin(4-14) treatment reduced the expression of cyclin E involved in the G(1)/S transition. Immunofluorescence microscopy showed that a lower γ-H2AX expression in lactoferricin(4-14)-treated cells, pointing to more efficient DNA repair. Thus, altogether our data show that lactoferricin(4-14) treatment has beneficial effects.