Peptide profiles and antioxidant capacity of extensive hydrolysates of milk protein concentrate.

Milk protein concentrate was hydrolyzed using one-step enzymatic hydrolysis. Both the peptide profiles and antioxidant activities of the resulting extensive hydrolysates of milk protein concentrate (EMPH) were analyzed using a peptidomics approach based on liquid chromatography-tandem mass spectrometry. The results demonstrated that the degrees of hydrolysis of the 4 EMPH by Alcalase-Protamex, Alcalase-Protease A 2SD, Alcalase-Flavorzyme, and Alcalase-ProteAXH were 12.02%, 16.85%, 15.87%, and 15.77%, respectively. Using size exclusion chromatography, 99.85% of the peptides in the Alcalase-Protease A 2SD hydrolysate were shown to have a molecular weight of <3 kDa. A total of 33 common peptides were identified in the EMPH by liquid chromatography-tandem mass spectrometry, 16 of which were identified as bioactive peptides using bioinformatics. The peptide profiles and the coverage of master proteins of the 4 EMPH were different. The EMPH also exhibited strong free radical scavenging capacity, as indicated by the results of the 1,1-diphenyl-2-picrylhydrazyl radical, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid), hydroxyl radical, and reducing power assays. The results of this study provided useful information on the peptide profiles and antioxidant activity of EMPH.

Ulinastatin Reduces the Severity of Intestinal Damage in the Neonatal Rat Model of Necrotizing Enterocolitis.

BACKGROUND Ulinastatin is a protease inhibitor derived from urine that has shown anti-inflammatory effects in human disease, including in sepsis. Necrotizing enterocolitis (NEC) is a common gastrointestinal disease in premature infants. Our aim was to explore the effects of ulinastatin on a neonatal NEC rat model. MATERIAL AND METHODS Forty-five neonatal rats were divided into 3 groups: normal control; NEC+sepsis-induced kidney injury (SIRS); NEC/SIRS+ulinastatin. The NEC/SIRS model was induced by injection of intraperitoneal saline, enteral formula feeding, hypoxia-hyperoxide, and cold stress exposure. The NEC/SIRS neonatal rats were perfused with ulinastatin at a dose of 10 000 u/kg/day. Giemsa staining and hematoxylin and eosin (H&E) were performed to evaluate the severity of intestinal damage. To assess intestinal cell apoptosis, we examined the expression of caspase-3 by TUNEL staining and western blot analysis. Intestinal levels of inflammatory cytokines (IL-1ß, IL-6, and TNF-alpha) were examined using ELISA assay. RESULTS Rats in the NEC treated with ulinastatin group had better physiological status and histological score compared to the NEC/SIRS group. Ulinastatin reduced NEC-induced weight loss. Macroscopic and microscopic morphology analyses showed that rats in the NEC treated with ulinastatin group had lower severity of intestinal damage compared to the NEC/SIRS group. TUNEL staining and caspase-3 expression detection results revealed that ulinastatin significantly inhibited intestinal cell apoptosis of NEC. Furthermore, ulinastatin decreased the intestinal levels of IL-1ß, IL-6, and TNF-alpha in NEC. CONCLUSIONS Ulinastatin could ameliorate the severity of intestinal damage in NEC and possess anti-apoptosis and anti-inflammation effects.

Comparison of proanthocyanidins A2 and B2 on IgE-reactivity and epitopes in Gly m 6 using multispectral, LC/MS-MS and molecular docking.

This study comparatively analyzed the changes in IgE-reactivity and epitopes in proanthocyanidins A2- (PA-Gly m 6) and B2-Gly m 6 (PB-Gly m 6) conjugates prepared by alkali treatment at 80 °C for 20 min. Similar to the western blot, ELISA also showed a higher reduced IgE-reactivity in PA-Gly m 6 (70.12 %) than PB-Gly m 6 (63.17 %). SDS-PAGE demonstrated that proanthocyanidins A2 caused more formation of >180 kDa polymers than proanthocyanidins B2. Multispectral analyses revealed that PA-Gly m 6 exhibited more structural alteration (e.g., a decrease of α-helical content and ANS fluorescence intensity) to unfold protein structure than proanthocyanidins B2, improving the accessibility to modify Gly m 6 for shielding or destroying conformational epitopes. LC/MS-MS revealed that PA-Gly m 6 conjugates had a lower abundance of allergens, peptides and linear epitopes than PB-Gly m 6 conjugates. Molecular docking showed that proanthocyanidins A2 and B2 reacted with Gln-317 and Asn-94 of epitopes, respectively. Overall, proanthocyanidins A2 is more effective than proanthocyanidins B2 to decrease the IgE-reactivity of Gly m 6 due to more shielding or destruction of conformational epitopes and lower content allergens and linear epitopes, which was attributed to more protein-crosslinks formation and structural changes in PA-Gly m 6 conjugates.

Multi-spectral and proteomic insights into the impact of proanthocyanidins on IgE binding capacity and functionality in soy 11S protein during alkali-heating treatment.

This study evaluated the impact of proanthocyanidins on immunoglobulin E (IgE) binding capacity, antioxidant, foaming and emulsifying properties in soy 11S protein following alkali treatment at 80 °C for 20 min. The formation of >180 kDa polymer was observed in the combined heating and proanthocyanidins-conjugation treatment sample (11S-80PC) rather than in the heating treated sample (11S-80) using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The structural analyzes demonstrated that 11S-80PC exhibited more protein unfolding than 11S-80. Heatmap analysis revealed that 11S-80PC had more alteration of peptide and epitope profiles in 11S than in 11S-80. Molecular docking showed that PC could well react with soy protein 11S. Liquid chromatography tandem MS analysis (LC/MS-MS) demonstrated that there was a 35.6 % increase in 11S-80, but a 14.5 % decrease in 11S-80PC for the abundance of total linear epitopes. As a result, 11S-80PC exhibited more reduction in IgE binding capacities than 11S-80 owing to more obscuring and disruption of linear and conformational epitopes induced by structural changes. Moreover, 11S-80PC exhibited higher antioxidant capacities, foaming properties and emulsifying activity than 11S-80. Therefore, the addition of proanthocyanidins could decrease allergenic activity and enhance the functional properties of the heated soy 11S protein.

The alteration of composition, conformation, IgE-reactivity and functional attributes in proanthocyanidins-soy protein 7S conjugates formed by alkali-heating treatment: Multi-spectroscopic and proteomic analyses.

This study assessed the alteration of IgE-reactivity and functional attribute in soy protein 7S-proanthocyanidins conjugates (7S-80PC) formed by alkali-heating treatment (pH 9.0, 80 °C, 20 min). SDS-PAGE demonstrated that 7S-80PC exhibited the formation of >180 kDa polymers, although the heated 7S (7S-80) had no changes. Multispectral experiments revealed more protein unfolding in 7S-80PC than in 7S-80. Heatmap analysis showed that 7S-80PC showed more alteration of protein, peptide and epitope profiles than 7S-80. LC/MS-MS demonstrated that the content of total dominant linear epitopes was increased by 11.4 % in 7S-80, but decreased by 47.4 % in 7S-80PC. As a result, Western-blot and ELISA showed that 7S-80PC exhibited lower IgE-reactivity than 7S-80, probably because 7S-80PC exhibited more protein-unfolding to increase the accessibility of proanthocyanidins to mask and destroy the exposed conformational epitopes and dominant linear epitopes induced by heating treatment. Furthermore, the successful attachment of PC to soy 7S protein significantly increased antioxidant activity in 7S-80PC. 7S-80PC also showed higher emulsion activity than 7S-80 owing to its high protein flexibility and protein unfolding. However, 7S-80PC exhibited lower foaming properties than 7S-80. Therefore, the addition of proanthocyanidins could decrease IgE-reactivity and alter the functional attribute of the heated soy 7S protein.

Effect of proanthocyanidins on protein composition, conformational structure, IgE binding capacities and functional properties in soybean protein.

This study was performed to determine the crosslinking formed by proanthocyanidins (PC) with respect to IgE binding capacities, functionality, structure and composition of soybean protein (SPI) following the alkali treatment at 60-100 °C. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed the formation of >180 kDa polymers, resulting from the formation of SPI-PC conjugates and protein cross-links. Structural analyses demonstrated that SPI-PC conjugates exhibited structural changes to unfold proteins and increase molecular flexibility. Liquid chromatography coupled to tandem mass spectrometry (LC/MS-MS) showed a decrease in unique protein and peptide numbers as well as major allergen and dominant epitopes abundance. When SPI was treated with PC under the alkali treatment at 80 °C, it exhibited a maximum reduction (68.8 %) in the immunoglobulin E (IgE) binding capacity and a maximum increase in DPPH radical scavenging activities (6.11-fold), ABTS + radical scavenging activities (4.80-fold), foaming stability (6.1 %) and emulsifying activity (27.3 %), compared to the control SPI. Overall, this study demonstrates that alkali treatment at 60-100 °C to form SPI-PC conjugates has potential applications for producing hypoallergenic soybean products with the desired functionality, most especially from alkali treatment at 80 °C. Moreover, the addition of PC pronouncedly alleviates the undesirable functional properties in heated SPI.

Associations of serum apolipoprotein A1, B levels and their ratio with blood pressure in Chinese adults with coronary artery disease.

OBJECTIVE: We examined the relationships of apolipoprotein A1 (ApoA1), ApoB levels and ApoB/A1 ratio with blood pressure (BP) in Chinese adults with coronary artery disease (CAD). METHODS: This cross-sectional study included 4921 adults with CAD. SBP, DBP, serum ApoA1 and ApoB levels were measured. The associations between Apo and BP were assessed by analyses of covariance. RESULTS: Serum ApoA1 was inversely associated with BP, whereas ApoB and the ApoB/A1 ratio exhibited positive associations with BP. For all subjects, a higher ApoA1 level was associated with lower SBP. Subjects in the fourth quartile for ApoA1 exhibited - 2.85 and - 2.63% lower DBP and mean arterial pressure (MAP), respectively than those in the third quartile. In contrast, higher ApoB and ApoB/A1 ratios were associated with higher SBP, DBP and MAP. The mean differences between ApoB quartiles 4 and 1 were 1.54% for SBP, 2.92% for DBP and 2.29% for MAP. The mean differences between the ApoB/A1 ratio quartiles 4 and 1 were 1.94% for SBP, 3.53% for DBP and 2.80% for MAP. In analyses stratified by gender, graded and inverse associations of ApoA1 with SBP, DBP and MAP were observed in both men and women, but positive associations were observed for ApoB and the ApoB/A1 ratio. Path analysis showed that BMI mediated the associations between ApoB and the ApoB/A1 ratio and SBP. CONCLUSIONS: In general, serum ApoA1 was inversely associated with BP in persons with CAD. In contrast, serum ApoB and the ApoB/A1 ratio were positively associated with BP, and these associations were mediated by BMI.

LncRNA DLX6-AS1 Promotes Malignant Phenotype and Lymph Node Metastasis in Prostate Cancer by Inducing LARGE Methylation.

Long non-coding RNAs (lncRNAs) have recently become recognized as crucial players in cancer cellular events including proliferation, migration, and invasion. Herein, we investigated the potential role of lncRNA DLX6-AS1 in prostate cancer cell malignant behaviors and lymph node metastasis. A differentially expressed lncRNA DLX6-AS1 and its downstream regulatory gene (LARGE) were predicted by analysis in silico. RT-qPCR and western blot analysis results demonstrated that DLX6-AS1 was highly expressed, but LARGE was poorly expressed in prostate cancer tissues and cells. The online website indicated that DLX6-AS1 negatively targeted LARGE expression, which was validated by Pearson correlation analysis and MSP. ChIP, RIP, and RNA pull-down assays further suggested that DLX6-AS1 downregulated LARGE expression through recruitment of DNMT1 to its promoter. We induced DLX6-AS1/LARGE overexpression or knockdown to examine their effects through Edu and Transwell assays, which revealed that DLX6-AS1 overexpression accelerated proliferation, invasion, and migration of prostate cancer cells, and that overexpression of LARGE rescued these effects. Tumors xenografts studies confirmed that DLX6-AS1 promoted lymph node metastasis by regulating LARGE, as evidenced by enhanced expression of MMP-9, uPAR, and cathepsin B. In summary, DLX6-AS1 stimulated prostate cancer malignant progression and lymph node metastasis by inducing DNMT1-mediated LARGE methylation, highlighting a potential therapeutic target against prostate cancer.

Preventive application of low molecular weight heparin ameliorates peripherally inserted central catheter-related venous thrombosis.

OBJECTIVE: Peripherally inserted central catheter (PICC) is being increasingly used in critical care settings. However, PICC is associated with various complications, particularly venous thrombosis. Our aim was to observe the effects of preventive application of low molecular weight heparin on venous thrombosis in a PICC model. METHODS: All rabbits were randomly divided into four groups: a control group, and low/medium/high concentration of low molecular weight heparin groups. All rabbits were injected prophylactically with normal saline or low molecular weight heparin once a day for 7 days. A PICC model was constructed. The pathologic changes of ear vein, anterior vena cava, and venous thrombosis were investigated using hematoxylin and eosin (H&E). Biochemical testing was performed including prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT). Serum D-dimer (D2D) and fibrinogen (FG) levels were detected using Enzyme-linked immunosorbent assay (ELISA). RESULTS: X-ray results showed that the PICC model was successfully constructed. H&E results showed that preventive application of low molecular weight heparin significantly ameliorated the pathologic damage to ear vein and anterior vena cava in the PICC model. Furthermore, we found that preventive application of low molecular weight heparin inhibited venous thrombosis in the model by H&E stain. Moreover, it significantly reduced serum FG and D2D levels in PICC model. Biochemical testing results showed that PT, APTT, and TT were significantly elevated in the PICC model. CONCLUSION: Our findings revealed that preventive application of low molecular weight heparin significantly ameliorates venous thrombosis in a PICC model.

Physiochemical properties and probiotic survivability of symbiotic oat-based beverage.

The objectives of this study were to develop a symbiotic oat-based beverage (SOB) and to analyze its physiochemical properties and the probiotic survivability. The beverage base was prepared by fermenting a mix containing oat flour (10%, w/w), sugar (4%, w/w), and inulin (1%, w/w) with a commercial Lactobacillus plantarum (0.003%, w/w) at 30 °C for 12 h. The SOB was formulated using the fermented oat base, sugar, stabilizers (pectin and λ-carrageenan), vitamin C, and citric acid. The beverage was analyzed for total solids (11.65 ± 0.22%), protein (0.58 ± 0.02%), fat (0.37 ± 0.02%), carbohydrate (10.70 ± 0.33%), ash (0.14 ± 0.01%), and dietary fiber (0.70 ± 0.05%). The pH value of the beverage was stable at about 3.60 during 7-week storage. Lactobacillus plantarum population in the beverage remained above 107 CFU/g throughout the storage. Oat-based beverage is a low fat and high dietary fiber symbiotic food.

Copeptin level in the early prediction of cardiorenal syndrome in rats.

Copeptin (CPP) has been considered as a useful marker for prediction of prognosis in heart diseases. However, CPP has not been investigated sufficiently in cardiorenal syndrome (CRS). The present study aimed to investigate the value of CPP level in predicting CRS in rats with partial nephrectomy combined with myocardial infarction (SNX + MI). A total of 60 male Sprague-Dawley rats were used to establish the CRS model by partial nephrectomy combined with MI. The rats were randomly divided into blank control (CK), SNX, MI and CRS groups. Changes in serum and urine CPP concentrations, hemodynamics, blood pressure, and renal function were examined 1-5 weeks after modeling. The predictive values of CPP in the occurrence of CRS in rats were evaluated using receiver operating characteristic (ROC) curve. The results showed that serum CPP in the CRS group in 1-5 weeks and urine CPP in 3 weeks after modeling increased significantly compared with the CK group. Also, serum B-type natriuretic peptide (BNP) in 1 and 3 weeks and urine BNP in 4-5 weeks after modeling increased significantly. No correlation was found between serum or urine CPP, BNP and BUN levels 1 week after modeling in the CRS group. The ROC curve analysis showed that the area under the curve of CRS predicted by serum CPP at 1 week was 0.908 with 56.59 pg/ml as the cutoff point, and its diagnostic sensitivity and specificity were 87.5 and 80.0%, respectively. To conclude, SNX + MI may be used to establish CRS rat model with cardiac and renal damage. Serum CPP may serve as a specific biomarker for the early prediction of CRS.