Identification of food-derived collagen peptides in human blood after oral ingestion of gelatin hydrolysates

In the present study, we identified several food-derived collagen peptides in human blood after oral ingestion of some gelatin hydrolysates. Healthy human volunteers ingested the gelatin hydrolysates (9.4-23 g) from porcine skin, chicken feet, and cartilage after 12 h of fasting. Negligible amounts of the peptide form of hydroxyproline (Hyp) were observed in human blood before the ingestion. After the oral ingestion, the peptide form of Hyp significantly increased and reached a maximum level (20-60 nmol/mL of plasma) after 1-2 h and then decreased to half of the maximum level at 4 h after the ingestion. Major constituents of food-derived collagen peptides in human serum and plasma were identified as Pro-Hyp. In addition, small but significant amounts of Ala-Hyp, Ala-Hyp-Gly, Pro-Hyp-Gly, Leu-Hyp, Ile-Hyp, and Phe-Hyp were contained.     

Angiotensin I-converting enzyme inhibitory activity of gelatin hydrolysates and identification of bioactive peptides

In this project we report on the angiotensin I-converting enzyme (ACE)-inhibitory activity of a bovine gelatin hydrolysate (Bh2) that was submitted to further hydrolysis by different enzymes. The thermolysin hydrolysate (Bh2t) showed the highest in vitro ACE inhibitory activity, and interestingly a marked in vivo blood pressure-lowering effect was demonstrated in spontaneously hypertensive rats (SHR). In contrast, Bh2 showed no effect in SHR, confirming the need for the extra thermolysin hydrolysis. Hence, an angiotensin I-evoked contractile response in isolated rat aortic rings was inhibited by Bh2t, but not by Bh2, suggesting ACE inhibition as the underlying antihypertensive mechanism for Bh2t. Using mass spectrometry, seven small peptides, AG, AGP, VGP, PY, QY, DY and IY or LY or HO-PY were identified in Bh2t. As these peptides showed ACE inhibitory activity and were more prominent in Bh2t than in Bh2, the current data provide evidence that these contribute to the antihypertensive effect of Bh2t.     

Assessing lipid coating of the human oral cavity after ingestion of fatty foods

The coating on the oral surface plays a significant role in mouthfeel perception, which is an important attribute governing perception of reduced-fat or low-fat food products. The aim of this work was to develop efficient methods to quantify the coating of the lipids covering the oral surface after ingestion of fatty foods. Three different approaches were assessed to investigate the in-mouth behavior of fluid foods in a subject. A first approach determined the mass of lipids retained in the oral cavity by rinsing it out. A second approach evaluated the thickness of the lipid coating on the tongue by fluorescence measurement using a dye incorporated in oil. Finally, a third approach measured local lipid thickness by adsorption of the lipids on a filter paper. All applied methods showed results in the same range. The thickness of the lipid coating was on average between 5 and 32 microm. The total lipid recoveries obtained by controlled rinses from the oral cavity were approximately 96 and 50% for single filter paper. A fast, simple, and quantitative method was developed to measure the thickness of the lipid coating on an oral surface after ingestion of fatty foods. This work presents the potential of the method to investigate in-mouth distribution and residues of lipids and establishes new avenues to study in-mouth behavior of food components and its influence on the sensory perception.     

Identification and characterization of topoisomerase II inhibitory peptides from soy protein hydrolysates

Topoisomerases are targets of several anticancer agents because their inhibition impedes the processes of cell proliferation and differentiation in carcinogenesis. With very limited information available on the inhibitory activities of peptides derived from dietary proteins, the objectives of this study were to employ co-immunoprecipitation to identify inhibitory peptides in soy protein hydrolysates in a single step and to investigate their molecular interactions with topoisomerase II. For this, soy protein isolates were subjected to simulated gastrointestinal digestion with pepsin and pancreatin, and the human topoisomerase II inhibitory peptides were co-immunoprecipitated and identified on a CapLC- Micromass Q-TOF Ultima API system. The inhibitory activity of these peptides from soy isolates toward topoisomerase II was confirmed using three synthetic peptides, FEITPEKNPQ, IETWNPNNKP,and VFDGEL, which have IC 50 values of 2.4, 4.0, and 7.9 mM, respectively. The molecular interactions of these peptides evaluated by molecular docking revealed interaction energies with the topoisomerase II C-terminal domain (CTD) (-186 to -398 kcal/mol) that were smaller than for the ATPase domain (-169 to -357 kcal/mol) and that correlated well with our experimental IC 50 values ( R (2) = 0.99). In conclusion, three peptides released from in vitro gastrointestinal enzyme digestion of soy proteins inhibited human topoisomerase II activity through binding to the active site of the CTD domain.     

beta-Conglycinins among sources of bioactives in hydrolysates of different soybean varieties that inhibit leukemia cells in vitro

Soybean is a complex matrix containing several potentially bioactive components. The objective was to develop a statistical model to predict the in vitro anticancer potential of soybean varieties based on the correlation between protein composition and bioactive components after simulated gastrointestinal enzyme digestion with their effect on leukemia mouse cells. The IC 50 values of the hydrolysates of soy genotypes (NB1-NB7) on L1210 leukemia cells ranged from 3.5 to 6.2 mg/mL. Depending on genotype, each gram of soy hydrolysates contained 2.7-6.6 micromol of total daidzein, 3.0-4.7 micromol of total genistein, 0.5-1.3 micromol of glycitein, 2.1-2.8 micromol of total saponins, 0.1-0.2 micromol of lunasin, and 0.1-0.6 micromol of Bowman-Birk inhibitor (BBI). The IC 50 values calculated from a partial least-squares (PLS) analysis model correlated well with experimental data ( R (2) = 0.99). Isoflavones and beta-conglycinin positively contributed to the cytotoxicity of soy on L1210 leukemia cells. Lunasin and BBI were potent L1210 cell inhibitors (IC 50 = 13.9 and 22.5 microM, respectively), but made modest contributions to the activity of defatted soy flour hydrolysates due to their relatively low concentrations. In conclusion, the data demonstrated that beta-conglycinins are among the major protein components that inhibit leukemia cell growth in vitro. Furthermore, it was feasible to differentiate soybean varieties on the basis of the biological effect of their components using a statistical model and a cell-based assay.     

Isolation and characterization of antioxidative peptides from gelatin hydrolysate of Alaska pollack skin

Gelatin extracted from Alaska pollack skin was hydrolyzed with serial digestions in the order of Alcalase, Pronase E, and collagenase using a three-step recycling membrane reactor. The fraction from the second step, which was hydrolyzed with Pronase E, was composed of peptides ranging from 1.5 to 4.5 kDa and showed high antioxidative activity. Two different peptides showing strong antioxidative activity were isolated from the hydrolysate using consecutive chromatographic methods including gel filtration on a Sephadex G-25 column, ion-exchange chromatography on a SP-Sephadex C-25 column, and high-performance liquid chromatography on an ODS column. The isolated peptides, P1 and P2, were composed of 13 and 16 amino acid residues, respectively; and both peptides contained a Gly residue at the C-terminus and the repeating motif Gly-Pro-Hyp. The antioxidative activities of the purified peptides were measured using the thiobarbituric acid method, and the cell viability was measured with MTT assay. The results showed that P2 had potent antioxidative activity on peroxidation of linoleic acid. Moreover, the cell viability of cultured liver cells was significantly enhanced by addition of the peptide. These results indicate that the purified peptide, P2, from gelatin hydrolysate of Alaska pollack skin is a natural antioxidant which has potent antioxidative activity.     

Comparison of carbon skeletal structures in black humic acids from different soil origins

Andosols are characterized by an abundance of black humic acids (HAs) belonging to Type A with a high content of aromatic carbon (C) in particular condensed aromatic C. Black HAs are also observed in other soils, such as Chernozems and the subsoil of paddy field, and extracted after washing with an acid or using chelating agent such as sodium pyrophosphate (Na₄P₂O₇). However, contribution of condensed aromatic structures to those soil HAs are unknown. To obtain the information about C skeletal structures of black HAs in soils other than Andosols, HAs were obtained from 2 Chinese Chernozem samples, 2 subsoil samples from Japanese paddy fields (Fulvisols), and a Rendzina-like soil (Cambisols) as well as an Andosol sample (reference) by successive extraction with 0.1 M NaOH (HAs₁) and 0.1 M Na₄P₂O₇ (HAs₂), and ¹³C nuclear magnetic resonance and X-ray diffraction 11-band profile analyses were applied. In the black HAs₂ from the non-Andosol samples, the proportion of C present as aromatic C, size of C layer planes, and relative C layer plane content ranged from 52 to 59%, 0.48 to 1.92 nm (mean size, 0.76–0.91 nm), and 58 to 100 AU (arbitrary unit) mg^?1, respectively, with a positive correlation between total C layer plane content and the degree of humification. Those ranges were similar to the distribution ranges of Andosols HAs₁ reported by our previous study.     

Identification of food-derived elastin peptide, prolyl-glycine (Pro-Gly), in human blood after ingestion of elastin hydrolysate

Elastin hydrolysate has apparent beneficial effects, and the food-derived peptide prolyl-glycine (Pro-Gly) is present in human blood after oral ingestion. Following ingestion of elastin hydrolysate (10 g/60 kg body weight) by healthy human volunteers, peripheral blood was used to prepare plasma samples from which peptides were extracted by solid phase extraction and fractionated by size-exclusion chromatography (SEC). Peptides in the SEC fractions were derivatized with phenyl isothiocyanate (PITC) and resolved by reversed phase (RP)-HPLC. Pro-Gly was the major food-derived elastin peptide, reaching a maximum (18 μM) at 30 min after ingestion, and decreasing to approximately 20% at 4 h after ingestion. Finally, in cell culture, levels of Pro-Gly in the medium above 0.1 μg/mL significantly enhanced elastin synthesis of normal human dermal fibroblasts (NHDF) without affecting the rate of cell proliferation.     

Preparation of antioxidant enzymatic hydrolysates from alpha-lactalbumin and beta-lactoglobulin. Identification of active peptides by HPLC-MS/MS

We have investigated the antioxidant activity of hydrolysates from whey proteins bovine alpha-lactalbumin (alpha-La) and beta-lactoglobulin A (beta-Lg A) by commercial proteases (pepsin, trypsin, chymotrypsin, thermolysin, and Corolase PP). Corolase PP was the most appropriate enzyme to obtain antioxidant hydrolysates from alpha-La and beta-Lg A (ORAC-FL values of 2.315 and 2.151 micromol of Trolox equivalent/mg of protein, respectively). A total of 42 peptide fragments were identified by HPLC-MS/MS in the beta-Lg A hydrolysate by Corolase PP. One of the sequences (Trp-Tyr-Ser-Leu-Ala-Met-Ala-Ala-Ser-Asp-Ile) possessed radical scavenging (ORAC-FL value of 2.621 micromol of Trolox equivalent/micromol of peptide) higher than that of butylated hydroxyanisole (BHA). Our results suggest that whey protein hydrolysates could be suitable as natural ingredients in enhancing antioxidant properties of functional foods and in preventing oxidation reaction in food processing.     

Simulations of ozone formation from different emission sources in sweden

An emission inventory concerning volatile organic compounds (VOC) and their emission profile linked to their sources in Sweden has been undertaken. The inventory has been used in model simulations to predict the ozone formation from different emission source categories in Sweden. The studies have been carried out using the IVL photochemical trajectory model for two types of air masses which describes clean and polluted air. In Sweden mobile sources contribute to 45 % by mass of the total national VOC emissions, 58 % of the NOx emissions and to at least 43 % of the ozone formation from national sources. In general, the ozone formation in Sweden is more dependent and sensitive to emissions of NOx rather than VOC.     

Obtention and characterization of phenolic extracts from different cocoa sources

The aim of this study was to evaluate several cocoa sources to obtain a rich phenol extract for use as an ingredient in the food industry. Two types of phenolic extracts, complete and purified, from different cocoa sources (beans, nibs, liquor, and cocoa powder) were investigated. UPLC-MS/MS was used to identify and quantify the phenolic composition of the extracts, and the Folin-Ciocalteu and vanillin assays were used to determine the total phenolic and flavan-3-ol contents, respectively. The DPPH and ORAC assays were used to measure their antioxidant activity. The results of the analysis of the composition of the extracts revealed that the major fraction was procyanidins, followed by flavones and phenolic acids. From the obtained results, the nib could be considered the most interesting source for obtaining a rich phenolic cocoa extract because of its rich phenolic profile content and high antioxidant activity in comparison with the other cocoa sources.     

Comparison of methods for measuring heavy metals and total phosphorus in soils contaminated by different sources

The relationships between the concentrations of zinc (Zn), lead (Pb), copper (Cu), nickel (Ni) and chromium (Cr) as measured by X-ray fluorescence analysis (XRF), aqua regia, and HNO₃ pressure digestion were studied in soil samples covering a wide range of heavy metal concentrations. The soils were contaminated by sewage sludge, exhaust depositions, river sediments of mining residues, and dump material. The question was addressed whether the source of heavy metals or other soil properties affect the relationship between these three methods. The aqua regia-digestible fraction of the five heavy metals reached on average 64% of the XRF-detectable content. The pressure accelerated HNO₃-digestible fraction of the five heavy metals was on average 71% of the XRF-detectable content; the respective phosphorus (P) fraction reached a median of 75%. This suggests that HNO₃ pressure digestion can also be used for characterizing the total P content of soils. Aqua regia extraction and HNO₃ pressure digestion gave similar values for Zn, Pb, and Cu, which dominate the heavy metal load of most contamination sources. Significantly higher Cr values were obtained by HNO₃ pressure digestion than by aqua regia extraction. Additionally, the Cr contents were affected by the source, e.g. sewage sludge had relatively high contents of aqua regia and HNO₃ pressure extractable contents in comparison to the XRF values. The element-specific relationships between the three methods were all highly significant. However, the respective multiple linear regression models were in most cases affected by soil organic carbon (C), in some cases by clay or soil pH.     

Isolation and identification of effective cellulolytic bacteria in composting process from different sources

Cellulolytic bacteria have a bio-activating role in the composting process. A study was carried out to isolate and identify cellulolytic bacteria from various sources. The isolates were cultured in the carboxymethyl cellulose (CMC) agar medium and incubated at 30°C for 3–7 days. Based on morphological characteristics of the isolates, maximum diameter of a clear zone around the colony and maximum cellulolytic activity, eight isolate were selected for further studies regarding composting experiments.

Molecular tests based on PCR amplification and sequencing of 16S rRNA gene of isolates showed the closest phylogenetic similarity with the species of Stenotrophomonas rhizophila DSM14405 (99.8%), Brevibacterium halotolerans DSM8802 (99.6%), Achromobacter marplatensis B2 (99.8%), Bacillus methylotrophicus CBMB205 (100%), Pseudomonas azotoformans IAM 1603 (99.7%), Bacillus sonorensis NBRC 101234 (99.8%), Bacillus subtilis KCTC 13429 (100%) and Ochrobactrum thiophenivorans DSM 7216 (99.3%). The study of the isolates impact on the composting of palm wastes in a randomized complete block design with 11 treatments in 3 replications showed that strain IB (B. methylotrophicus) caused a significant decrease in C:N ratio (58%). The increasing of microbial respiration compared with control after 30 days incubation at 37°C showed that the B. methylotrophicus strain IB with cellulolytic characteristics can be applied to hydrolysis of cellulosic biomass in the composting processes.     

Comparison of potential potassium leaching associated with organic and inorganic potassium sources in different arable soils in China

Potassium (K) leaching is detrimental to the maintenance of sustainable arable soil K fertility,especially in low-K fixation soils.It is not known whether the application of inorganic fertilizers with lower K mobility or crop straw can reduce potential K leaching in low-K fixation arable soils.The potential K leaching of 14 representative arable soils with different K fixation capacities in China was evaluated with or without the addition of K under two rainfall intensities (90 and 225 mm),and then potential K leaching was assessed in relation to five K sources (KCl,K₂SO₄,KH₂PO₄,maize (Zea mays L.) straw,and rice (Oryza sativa L.) straw).Without K addition,K leaching mainly occurred in sandy soils at 90 mm of rainfall and in soils with greater organic matter at225 mm of rainfall.With K addition,the leaching percentage of exogenous K ranged from 0.6%to 11.6%at 90 mm of rainfall and 1.2%to 21.2%at 225 mm of rainfall.The greatest K leaching occurred in soils with fewer K-bearing minerals and lower pH at both rainfall intensities.In most cases,KH₂PO₄,which has lower K mobility,markedly reduced K leaching in both high-and low-K leaching soils at the two rainfall intensities.Maize and rice straw reduced K leaching only in soils with high K leaching,regardless of rainfall amount,whereas more K was leached in soils with lower K leaching at high rainfall intensity.In conclusion,KH₂PO₄ and straw should be preferred for reducing K leaching in low-K fixation arable soils.     

Acclimation and response of algal communities from different sources to zinc toxicity

Algae from two different sources, Peoria and Farmington, Illinois, exhibited distinctively different response patterns to Zn toxicity. This study was undertaken to find the cause of the variation. The first experiment confirmed that algae, not water sample, were the cause of the variation in response. Additional experiments proved that when the Peoria algae were acclimated to a low Zn environment, they behaved like the Farmington algae. The Farmington algae behaved like the Peoria algae if they were allowed to acclimate in water samples containing higher Zn concentrations. The results indicate that the toxic response of an algal community is greatly dependent upon specific environment and algal acclimation to it, regardless of the origin of algal sources.     

Antagonistic and synergistic effects of fungal and bacterial growth in soil after adding different carbon and nitrogen sources

The effect of adding easily available and more complex carbon sources, with and without nitrogen, on fungal and bacterial growth and activity in soil were studied in the laboratory. Total microbial activity was estimated by measuring respiration, fungal growth with the acetate-in-ergosterol incorporation technique and bacterial growth with the thymidine and leucine incorporation techniques. The substrate additions consisted of glucose and cellulose, with and without nitrogen (as ammonium nitrate), and gelatine. The microbial development was followed over a 2-month period. The respiration rate increased within a few days after adding glucose, with and without nitrogen, and gelatine, initially by more than 10 times, but after 2 months no differences were seen compared with the control. Bacterial growth estimated with the thymidine and leucine incorporation techniques gave similar results. Adding glucose with nitrogen, or gelatine, increased bacterial growth within a few days up to 10 times, but even after 2 months of incubation bacterial growth rates were still about 5 times higher than in the control. Adding only glucose increased bacterial growth rates by about twice over the whole incubation period. Fungal growth rates especially increased after adding cellulose and nitrogen, although a minor increase was found after adding cellulose alone. Fungal growth rates started to increase after 10 days of incubation with cellulose. There were indications of synergistic effects in that bacterial growth increased after the fungi had started to grow after adding cellulose. Treatments resulting in high bacterial growth rates (adding easily available carbon sources) led to decreased fungal growth rates compared with the control, indicating antagonistic effects of bacteria.     

Honeys from different floral sources as inhibitors of enzymatic browning in fruit and vegetable homogenates

Honeys from different floral sources were evaluated for their antioxidant content and for their ability to inhibit enzymatic browning in fruits and vegetables. Antioxidant contents of honeys vary widely from different floral sources, as do their abilities to protect against enzymatic browning. Polyphenol oxidase (PPO) activity was reduced over a range of approximately 2-45% in fruit and vegetable homogenates, corresponding to a reduction in browning index by 2.5-12 units. Soy honey was particularly effective when compared to clover honey, which had a similar antioxidant content. When compared to commercial inhibitors of browning, honeys were less effective; however, in combination they added to the effectiveness of metabisulfite and ascorbic acid. Honey has great potential to be used as a natural source of antioxidants to reduce the negative effects of PPO browning in fruit and vegetable processing.     

不同蛋白源有机肥分解过程中组成性质变化特征

不同商品有机肥在组成和性质上存在极大差异,决定着其在土壤环境中分解规律的不同.采用元素分析、热重分析(TG)、傅里叶变换红外光谱(FTIR)等光谱技术,分析了3种蛋白源的商品(豆粕、骨粉、虾肽)有机肥分解过程中物质组成、官能团组成、元素组成以及理化性质的变化.经过12个月的矿化分解,3种有机肥的pH值、CEC和灰分含量...     

Stereospecific analysis of phospholipid classes in skeletal muscle from rats fed different fat sources

The fatty acid (FA) and dimethylacetal profiles of the sn-1 and sn-2 positions of different phospholipid (PL) classes from skeletal muscle of rats as affected by dietary FA profiles were studied. Rats were fed either a control diet, an olive oil-enriched diet, or a sunflower oil-enriched diet. The FA composition of both positions of the studied PL classes was affected by diet to different extents. The FA composition of the sn-2 position of phosphatidylserine was the most influenced by diet, while phosphatidylinositol was less affected by dietary modification. The FA profile of phosphatidylcholine reflected consumed FA better than any other studied PL. Thus, olive oil rats showed higher oleic acid (C18:1 n-9) contents in both positions of phosphatidylcholine, and sunflower oil rats had higher proportions of arachidonic acid (C20:4 n-6) in the sn-1 position of this PL class. Dimethylacetals were scarcely affected by diet, and only the dimethylacetal composition of phosphatidylethanolamine showed significant modifications.