Relationship between functional properties and structure of ovalbumin

The effects of ovalbumin (OVA) denaturation using urea, guanidinium chloride (GdnHCl), sodium dodecyl sulphate (SDS), cetylpyridinium chloride (CPC), 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), and 5 different cationic detergents with various side chains, HCl, and CH₃COOH were observed. Progressive unfolding in ovalbumin was measured as a function of fluorescent light intensity, peak response and shift in the maximum of emission. Kinetic measurements demonstrated that the rate of denaturation usually followed a double exponential decay pattern, but at small concentrations of urea and acids first-order reaction was indicated. The reversibility of the unfolding-folding transitions was confirmed from tryptophan fluorescence and circular dichroism (CD) measurements. Differences in secondary structure were observed and changes of-helical content were calculated. Polyacrylamide gel electrophoresis (PAGE) with and without sodium dodecyl sulphate (SDS-PAGE) showed differences in the structure of native and denatured ovalbumin. Native protein samples in PAGE demonstrated smaller number and larger mobilities of subunits than denatured ones with different reductants, such as SDS and 2-mercaptoethanol (2 ME). Scanning of SDS protein patterns showed the appearance of aggregated forms in region of 45 kD.     

The nutritional and metabolic indices in rats fed cholesterol-containing diets supplemented with durian at different stages of ripening

The aim of this investigation was to assess the nutritional and health properties of Mon Thong durian cultivar at different stages of ripening. The assessment was carried out in vitro and in vivo. The contents of dietary fibers, minerals and trace metals at different stages of ripening were comparable. Total polyphenols (mgGAE/100 g FW) and flavonoids (mg CE/100 gFW) in ripe durian (358.8 +/- 31.4 and 95.4 +/- 9.3) were significantly higher (p < 0.05) than in mature (216.1 +/- 1 and 39.9 +/- 3.8) and overripe (283.3 +/- 26.2 and 53.5 +/- 4.9). Antioxidant capacity (muMTE/100 g FW) in total polyphenol extracts of ripe durian measured by 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and [2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid)] (ABTS) assays (259.4 +/- 23.6 and 2341.8 +/- 93.2) were significantly higher (p < 0.05) than that of mature (151.6 +/- 15.2 and 1394.6 +/- 41.5) and overripe (201.7 +/- 19.4 and 1812.2 +/- 61.4) samples. The correlation coefficients between the bioactive compounds in different stages of ripening and their antioxidant capacities were high (R;{2} = 0.99). Then 35 male Wistar rats were divided into 5 dietary groups each of 7 and named Control, Chol, Chol/Mature, Chol/Ripe and Chol/Overripe. During 30 days of the experiment the rats of all 5 groups were fed basal diet (BD), which included wheat starch, casein, soybean oil, vitamin and mineral mixtures. The rats of the Control group were fed a BD only. To the BD of the Chol group was added 1% of cholesterol. The BD of the Chol/Mature, Chol/Ripe and Chol/Overripe groups was supplemented with 1% of cholesterol and 5% of the mature, ripe and overripe durian as freeze-dried powder, respectively. Diets containing ripe and to a lesser degree mature and overripe durian significantly hindered the rise in plasma lipids and also hindered a decrease in plasma antioxidant activity. The nitrogen retention in rats of the Chol/Ripe group was significantly higher (63.6%, P < 0.05) than in other diet groups and the level of the plasma glucose remained normal. A decrease in fibrinogen fraction with ripe durian included in rat's diets was shown by electrophoretic separation. These changes were detected mostly in the low molecular weight proteins of rat's serum. Histological examination of aorta showed only slight differences in the tissue. In conclusion, ripe durian contains higher quantity of bioactive compounds, has higher antioxidant capacity and nutritional value. It positively affects the plasma lipid profile, the plasma glucose and the antioxidant activity in rats fed cholesterol enriched diets. Therefore, the ripe durian supplemented diet could be beneficial for patient suffering from hypercholesterolemia and diabetes mellitus. <     

Stability of Collagen During Denaturation

The stability of calf skin collagen (CSC) type I during thermal and chemical denaturation in the presence of glycerol was investigated. Thermal denaturation of type I collagen was performed in the presence of glycerol or in combination with urea and sodium chloride. The denaturation curves obtained in the presence of urea or sodium chloride retained their original shape without glycerol. These curves were shifted upward proportionally to the glycerol concentration in the reaction medium. This means that glycerol and the denaturants act independently. The explanation is based on the difference in the mechanism of their action on the collagen molecule.     

Oat (Avena sativa L.) and amaranth (Amaranthus hypochondriacus) meals positively affect plasma lipid profile in rats fed cholesterol-containing diets

Cereals are an important part of diets for hypercholesterolemic patients. However, some of these patients are allergic to these natural products. The purpose of the current study was to compare oatmeal with equal in nutritional values two allergy-free amaranth meals to determine whether this pseudocereal can be a substitute for allergic to cereals individuals. The total phenols of the samples were determined with the Folin-Chocalteu reagent, anthocyanins, and flavonoids spectrophotometrically. The antioxidant activities were estimated with nitric oxide scavenging radical (NO) and by beta-carotene bleaching (beta-carotene). It was found that the contents of different protein fractions, antioxidant compounds, and the antioxidant activities of oatmeal were significantly higher than those of the two amaranth samples. The results of kinetic reactions showed that samples differed in their capacities to quench these radicals, and oats have shown more antioxidant activity than amaranth. High correlation was observed between antioxidant activities and phenols (R(2) = 0.99). In the in vivo part of the investigation, 60 male Wistar rats were divided into five diet groups of 12 animals each; these groups were designated as Control, Chol, Chol/Oat, Chol/AmarI, and Chol/AmarII. The rats of the Control group were fed basal diet (BD) only. To the BD of the four other groups were added the following: 1% of cholesterol (Chol), 10% of oat meal and 1% of cholesterol (Chol/Oat), 10% of amaranth I meal, and 1% of cholesterol (Chol/AmarI) and 10% of amaranth II meal and 1% of cholesterol (Chol/AmarII). After 32 days of different feeding, diets supplemented with oat meal and, to lesser degree, with amaranth I and amaranth II hindered the rise in the plasma lipids: a) TC: 3.14 vs. 4.57 mmol/L, - 31.3%; 3.31 vs. 4.57 mmol/L - 27.6%; and 3.40 vs. 4.57, - 25.6%, respectively b) LDL-C: 1.69 vs. 3.31 mmol/L, - 49.9%; 2.05 vs. 3.31 mmol/L, - 38.1%; and 2.16 vs. 3.31 mmol/L, - 34.8%, respectively; c) TG: 0.73 vs. 0.88 mmol/L, - 17.1%; 0.75 vs. 0.88 mmol/L, - 14.8%; and 0.79 vs. 0.88 mmol/L, -10.2%, respectively. The HDL-PH was increased as follows: 0.79 vs. 0.63 mmol/L, -25.3%; 0.75 vs. 0.63 mmol/L, -23.0%; and 0.71 vs. 0.63 mmol/L, -12.7% for the Chol/Oat, Chol/AmarI and Chol/AmarII, respectively. No significant changes in the concentrations of HDL-C and TPH were found; however the HDL-C in the Chol/Oat group was slightly higher than in other groups. No changes in the Control group were registered. In conclusion, oat and amaranth meals positively affect plasma lipid profile in rats fed cholesterol-containing diets. The degree of this positive influence is directly connected to the contents of the bioactive components and the antioxidant activities of the studied samples. It is suggested that amaranth could be a valuable substitute for hypercholesterolemic patients allergic to cereals.     

Comparative content of total polyphenols and dietary fiber in tropical fruits and persimmon

Recent studies have shown that dietary fiber and polyphenols of vegetables and fruits improve lipid metabolism and prevent the oxidation of low density lipoprotein cholesterol (LDL-C), which hinder the development of atherosclerosis. The goal of this study was to measure the total polyphenol and dietary fiber contents of some tropical fruits (i.e., pineapple, wax apple, rambutan, lichi, guava, and mango) and compare the results to the content of these substances in the better characterized persimmon. It was found that lichi, guava, and ripe mango (cv. Keaw) have 3.35, 4.95, and 6.25 mg of total polyphenols in 100 g fresh fruit, respectively. This is significantly higher than in persimmon, pineapple, wax apple, mature green mango, and rambutan [P < 0.0005 for pineapple (Smooth Cayene variant), wax apple, persimmon, rambutan, mature green mango (cv. Keaw); the value of P < 0.001 is found only for pineapple (Phuket, Queen variant)]. The same relationship was observed for the contents of gallic acid and of dietary fiber. It can be supposed that among the studied fruit, lichi, guava, and ripe mango may be preferable for dietary prevention of atherosclerosis.     

Quantitative analysis of heterocyclic amines in urine by liquid chromatography coupled with tandem mass spectrometry

A sensitive, reproducible, and rapid analytical method for the analysis of trace-level heterocyclic amines (HCAs) that are expected to have high levels of human exposure was developed. Liquid–liquid extraction (LLE) with dichloromethane (DCM) followed by solid-phase extraction (SPE) was carried out. Liquid extraction with DCM under basic conditions was efficient in extracting HCAs from urine samples. For further purification, mixed mode cationic exchange (MCX) cartridges were applied to eliminate the remaining interferences after liquid extraction. Separation and quantification were performed by liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) in selected reaction monitoring (SRM) mode. The overall recoveries ranged between 71.0% and 113.6% with relative standard deviations (RSDs) of 5.1% to 14.7% for the entire procedure. The limits of detection (LODs) and limits of quantification (LOQs) of the proposed analytical method were in the ranges of 0.04 to 0.10 ng/ml and 0.15 to 0.36 ng/ml, respectively. This method was applied to the analysis of monitoring in urine samples for Korean school children, and the results demonstrated that the method can be used for the trace determination of HCAs in urine samples.     

Structure characterization of human serum proteins in solution and dry state.

The present report describes application of advanced analytical methods to establish correlation between changes in human serum proteins of patients with coronary atherosclerosis (protein metabolism) before and after moderate beer consumption. Intrinsic fluorescence, circular dichroism (CD), differential scanning calorimetry and hydrophobicity (So) were used to study human serum proteins. Globulin and albumin from human serum (HSG and HSA, respectively) were denatured with 8 m urea as the maximal concentration. The results obtained provided evidence of differences in their secondary and tertiary structures. The thermal denaturation of HSA and HSG expressed in temperature of denaturation (Td, degrees C), enthalpy (DeltaH, kcal/mol) and entropy (DeltaS kcal/mol K) showed qualitative changes in these protein fractions, which were characterized and compared with fluorescence and CD. Number of hydrogen bonds (n) ruptured during this process was calculated from these thermodynamic parameters and then used for determination of the degree of denaturation (%D). Unfolding of HSA and HSG fractions is a result of promoted interactions between exposed functional groups, which involve conformational changes of alpha-helix, beta-sheet and aperiodic structure. Here evidence is provided that the loosening of the human serum protein structure takes place primarily in various concentrations of urea before and after beer consumption (BC). Differences in the fluorescence behavior of the proteins are attributed to disruption of the structure of proteins by denaturants as well as by the change in their compactability as a result of ethanol consumption. In summary, thermal denaturation parameters, fluorescence, So and the content of secondary structure have shown that HSG is more stable fraction than HSA.