Strukturanalyse der stark gekrümmten Caveolenmembran

In dieser Arbeit war die Gefrierbruch-Immunogoldmarkierung die bevorzugt angewendete Methode. Um die Struktur der Caveolen und die Verteilung des Caveolins mit Hilfe der Gefrierbruch-Immunogoldmarkierung genau untersuchen zu können, war zunächst eine Optimierung dieser Methode essentiell. Es konnte gezeigt werden, dass die Markierungsrate von Caveolin-1 an den Gefrierbruch-Replika entscheidend von der Schichtdicke der aufgedampften Platinschicht beeinflusst wird. Die Markierungsraten von Caveolin-1 konnten durch das Aufdampfen einer dünneren Platinschicht von 1,5 nm Stärke und durch die Variation der Bedampfungsreihenfolge, bei der zuerst die Kohlenstoffschicht und als zweites die Platinschicht aufgedampft wurde, signifikant gesteigert werden. Darüber hinaus spielt wahrscheinlich die Membrantopologie des zu markierenden Proteins eine Rolle für die Effizienz der Immunmarkierung an den Gefrierbruch-Replika

Effect of a regular consumption of traditional and roasted oat and barley flakes on blood lipids and glucose metabolism–A randomized crossover trial

Background Regular consumption of the soluble dietary fiber β-glucan is associated with decreased total cholesterol (TC), low-density lipoprotein (LDL) cholesterol and blood glucose. Barley and oat flakes as natural sources of β-glucan were roasted to improve sensory quality. The aim of this study was to investigate whether roasting of barley and oat flakes changes the physiological impact of the β-glucan-rich flakes on glucose and lipid metabolism. Method A five-armed randomized crossover trial design was used. The intervention study was conducted from May 2018 to May 2019 and included 32 healthy subjects with moderately increased LDL cholesterol (≥2.5 mmol/L). During the 3-week intervention periods, 80 g of roasted or traditional barley or oat flakes, or four slices of white toast bread per day were consumed for breakfast. At the start and the end of each intervention, fasting and postprandial blood was taken. The intervention periods were separated by 3-week wash-out periods. Results During the interventions with the cereal flakes, TC and LDL cholesterol concentrations were significantly reduced compared to baseline values by mean differences of 0.27–0.33 mmol/L and 0.21–0.30 mmol/L, respectively ( p < 0.05), while high-density lipoprotein (HDL) cholesterol was only reduced after the intervention with barley flakes ( p < 0.05). After the intervention period with toast, TC and HDL cholesterol increased ( p < 0.05). The fasting levels of triglycerides, fasting blood glucose and insulin did not change in any group. The effects of traditional and roasted varieties on blood lipids did not differ between the groups. Conclusion The regular consumption of traditional or roasted barley and oat flakes contributes to the management of cardiovascular diseases by improving TC and LDL cholesterol. Clinical trial registration https://clinicaltrials.gov/ct2/show/NCT03648112 , identifier NCT03648112

Chemopreventive Potential of Raw and Roasted Pistachios Regarding Colon Carcinogenesis

Pistachios are rich in health-promoting bioactive compounds such as B vitamins, γ-tocopherol, polyphenols and dietary fiber, which could contribute to the reduction of colon cancer risk in terms of chemoprevention (Fischer, S.; Glei, M. Health-Potential of Nuts. Ernaehrungs Umsch. Int. 2013, 60, 206–215.). Since pistachios are often consumed roasted, the present study aims at investigating the influence of different roasting conditions (RC) on potential chemopreventive effects of pistachios in colon adenoma cells such as growth and apoptosis, genotoxic- and anti-genotoxic effects and modulation of gene expression of detoxifying enzymes (CAT, SOD2, GPx1, and GSTP1). Fermentation supernatants (FS) were obtained from raw and roasted (RC1 = 141 °C/25 min, RC2 = 160 °C/15 min and RC3 = 185 °C/21 min) pistachios after in vitro fermentation. FS of pistachios significantly reduced LT97 cell growth in a time- and dose-dependent manner. Compared to the blank control, pistachio FS (2.5%) led to a significant average reduction of H2O2-induced DNA damage (1.5-fold). Levels of CAT mRNA were significantly increased (1.3-fold, on average for 5% FS). Pistachio FS (5%) significantly increased the number of early apoptotic cells (up to 2.1-fold) and levels of caspase-3 activities (up to 6.9-fold). The present results confirm a chemopreventive potential of pistachios, which is mediated by growth inhibition, induction of apoptosis and anti-genotoxic effects, as well as induction of CAT. These effects remain mostly unaffected by roasting

Potential Role of ROS in Butyrate- and Dietary Fiber-Mediated Growth Inhibition and Modulation of Cell Cycle-, Apoptosis- and Antioxidant-Relevant Proteins in LT97 Colon Adenoma and HT29 Colon Carcinoma Cells

The aim of the present study was to examine whether reactive oxygen species (ROS) contribute to chemopreventive effects of fermentation supernatants (FS) of different dietary fibers (Synergy1®, oat-, barley-, yeast β-glucan, Curdlan) and butyrate as a fermentation metabolite. LT97 and HT29 cells were treated with butyrate and FS alone or with N-acetyl-cysteine (NAC) and their impact on ROS formation, cell growth, and protein expression (Cyclin D2, p21, PARP, Bid, GPx2) was investigated. Butyrate and FS significantly decreased cell growth. ROS levels were significantly increased, particularly in LT97 cells, while co-treatment with NAC decreased ROS formation and growth inhibitory effects in both cell lines. After treatment with butyrate and FS, Cyclin D2 expression was reduced in LT97 cells and p21 expression was increased in both cell lines. Levels of full-length PARP and Bid were decreased, while levels of cleaved PARP were enhanced. GPx2 expression was significantly reduced by fiber FS in HT29 cells. A notable effect of NAC on butyrate- and FS-modulated protein expression was observed exclusively for PARP and Bid in HT29 cells. From the present results, a contribution of ROS to growth inhibitory and apoptotic effects of butyrate and FS on LT97 and HT29 cells cannot be excluded

In Vitro Digested Nut Oils Attenuate the Lipopolysaccharide-Induced Inflammatory Response in Macrophages

Nut consumption is known for its health benefits, in particular in inflammatory diseases. A possible mechanism for these effects could be their beneficial fatty acid composition. Nuts mainly contain mono- and polyunsaturated fatty acids, which have anti-inflammatory properties. However, studies investigating the effects of nut extracts on inflammatory processes on the molecular level are rare. We therefore prepared oily nut extracts after in vitro digestion and saponification of the fat-soluble constituents. Besides chromatographic analysis, cell culture experiments were performed using murine macrophages (RAW264.7) to study the capacity of different nut extracts (hazelnut, almond, walnut, macadamia, and pistachio) to modulate inflammatory processes. Oleic acid was the main fatty acid in hazelnut, almond, macadamia, and pistachio extracts. Both oily nut extracts and pure oleic acid significantly reduced the LPS-induced expression of iNos, Cox2, Tnf&#945;, Il1&#946;, and Il6 mRNAs. iNos protein expression was down-regulated followed by reduced nitric oxide formation. Thus, nut extracts at concentrations achievable in the digestive tract inhibit the expression and formation of inflammatory mediators in macrophages. Hence, a beneficial contribution of nut consumption to inflammatory diseases can be assumed. We are convinced that these results provide new insights on the molecular mechanisms involved in the health-beneficial effects of nuts

Influence of roasting on the chemopreventive potential of <i>in vitro</i> fermented almonds in LT97 colon adenoma cells

<p>The consumption of almonds may contribute to the prevention of colon cancer due to their unique composition of health promoting compounds. Since almonds are often consumed roasted, the impact of different roasting (R) conditions (R1 = 139.2 °C/25 min, R2 = 161.5 °C/20 min and R3 = 170.8 °C/15 min) on chemopreventive effects of <i>in vitro</i>-fermented almonds was analysed in LT97 colon adenoma cells. Fermentation supernatants (FS) of raw and roasted almonds had no genotoxic effects. FS obtained from raw or mildly roasted almonds (R1) significantly increased mRNA levels of <i>CAT</i> (4.6-fold), <i>SOD2</i> (5.6-fold) and <i>GSTP1</i> (3.9-fold) but not of <i>GPx1</i>. FS of almonds significantly reduced the growth of LT97 cells in a time- and dose-dependent manner. Treatment with 5% almonds FS increased the number of early apoptotic cells (17.4%, on average) and caspase-3 activity (4.9-fold, on average). The results indicate a chemopreventive potential of <i>in vitro</i>-fermented almonds which is largely independent of the roasting process.</p

Use of the β-Glucan-Producing Lactic Acid Bacteria Strains <i>Levilactobacillus brevis</i> and <i>Pediococcus claussenii</i> for Sourdough Fermentation—Chemical Characterization and Chemopreventive Potential of In Situ-Enriched Wheat and Rye Sourdoughs and Breads

The aim of the present study was to examine β-glucan production and the potential prebiotic and chemopreventive effects of wheat and rye sourdoughs and breads generated with wild-type and non-β-glucan-forming isogenic mutant strains of Levilactobacillus brevis and Pediococcus claussenii. Sourdough and bread samples were subjected to in vitro digestion and fermentation. Fermentation supernatants (FS) and pellets (FP) were analyzed (pH values, short-chain fatty acids (SCFA), ammonia, bacterial taxa) and the effects of FS on LT97 colon adenoma cell growth, viability, caspase-2 and -3 activity, genotoxic and antigenotoxic effects and on gene and protein expression of p21, cyclin D2, catalase and superoxide dismutase 2 (SOD2) were examined. Concentrations of SCFA were increased and concentrations of ammonia were partly reduced in the FS. The relative abundance of Bifidobacteriaceae was increased in all FPs. Treatment with FS reduced the growth and viability of LT97 cells and significantly increased caspase-2 and -3 activities without exhibiting genotoxic or antigenotoxic effects. The p21 mRNA and protein levels were increased while that of cyclin D2 was reduced. Catalase and SOD2 mRNA and protein expression were marginally induced. The presented results indicate a comparable chemopreventive potential of wheat and rye sourdoughs and breads without an additional effect of the formed β-glucan