Investigation of the peptides with calcium chelating capacity in hydrolysate derived from spent hen meat.

Calcium peptide chelates are developed as efficient supplements for preventing calcium deficiency. Spent hen meat (SHM) contains a high percentage of proteins but is generally wasted due to the disadvantages such as hard texture. We chose the underutilized SHM to produce peptides to bind calcium by proteolysis and aimed to investigate chelation between calcium and peptides in hydrolysate for a sustainable purpose. The optimized proteolysis conditions calculated from the result of response surface methodology for two-step hydrolysis were 0.30% (wenzyme/wmeat) for papain with a hydrolysis time of 3.5 h and 0.18% (wenzyme/wmeat) for flavourzyme with a hydrolysis time of 2.8 h. The enzymatic hydrolysate (EH) showed a binding capacity of 63.8 ± 1.8 mg calcium/g protein. Ethanol separation for EH improved the capacity up to a higher value of 68.6 ± 0.6 mg calcium/g protein with a high association constant of 420 M-1 (25°C) indicating high stability. The separated fraction with a higher amount of Glu, Asp, Lys, and Arg had higher calcium-binding capacity, which was related to the number of ─COOH and ─NH2 groups in peptide side chains according to the result from amino acid analysis and Fourier transform infrared spectroscopy. Two-step enzymatic hydrolysis and ethanol separation were an efficient combination to produce peptide mixtures derived from SHM with high calcium-binding capacity. The high percentage of hydrophilic amino acids in the separated fraction was concluded to increase calcium-binding capacity. This work provides foundations for increasing spent hen utilization and developing calcium peptide chelates based on underutilized meat.

Green tea polyphenols bind to soy proteins and decrease the activity of soybean trypsin inhibitors (STIs) in heated soymilk.

The interaction between epigallocatechin gallate (EGCG) and soy proteins at room temperature (25 °C) and after heating at 100 and 121 °C, and their effects on the inactivation of soybean trypsin inhibitors (STIs) in soymilk were investigated. The results of the nitroblue tetrazolium (NBT) staining assay showed that soy proteins can covalently bind to EGCG. The α/α' and A subunits in heated soymilk preferred to bind to EGCG because of their soluble state. More thiols were trapped when EGCG was added before thermal processing, and the free amino groups were depleted more with EGCG addition after heating. Circular dichroism and fluorescence spectroscopy showed that EGCG addition before or after heating induced different secondary and tertiary structural changes for soy proteins. The exposed aromatic amino acids preferred to react with EGCG before protein aggregation in the heating process. The random coil of soymilk proteins increased more when EGCG was added in soymilk after heating, resulting in more disordered structures in protein conformation. The binding between EGCG and soy proteins promoted protein aggregation, which was confirmed by the particle size distribution and gel electrophoresis. The trypsin and chymotrypsin inhibitory activity (TIA and CIA) in soymilk significantly reduced to 693 U mL-1 and 613 U mL-1, respectively, under the conditions of 2 mM EGCG addition after 100 °C heating for 10 min (p < 0.05). Consequently, the influence of EGCG on STI inactivation in soymilk only worked when EGCG was added after heating.

Pepsin Diffusivity and In Vitro Gastric Digestion of Soymilk as Affected by Binding of Tea Polyphenols to Soy Proteins.

This study aimed to investigate the effect of tea polyphenol extract (TPE) on the in vitro gastric digestion of soymilk. Fluorescence recovery after photobleaching was applied to measure pepsin diffusivity in soymilk. The characteristics of soymilk digesta were evaluated by gel electrophoresis, degree of hydrolysis (DH), molecular weight distribution, free amino acid analysis, particle size, antioxidant capacity, and trypsin/chymotrypsin inhibitor activity (TIA/CIA). The binding between soy proteins and tea polyphenols could significantly impair in vitro gastric digestion of soymilk by decreasing pepsin diffusivity from 91.3 to 70.3 µm2/s and DH from 17.13 to 13.93% with 1.2 mg/g TPE addition. Soymilk with 0.6 mg/g TPE addition exhibited low TIA/CIA and a strong antioxidant capacity in gastric digesta, which might be good for the following intestinal digestion. A better understanding of the effect of polyphenol on the digestion of protein-based food may be beneficial to innovation in food manufacturing.

Isolation, purification, structure characterization of a novel glucan from Huangshui, a byproduct of Chinese Baijiu, and its immunomodulatory activity in LPS-stimulated THP-1 cells.

Huangshui (HS) has attracted considerable attention for the utilization of aroma compounds and microorganisms containing presently. However, little work has been done on biological polysaccharides in HS. In this study, a novel water-eluted HS polysaccharide, HSP-W, was identified as an α-D-glucan with a Mw of 166.00 kDa. It was consisted of a 1,4 linked α-D-Glcp backbone with the substitution at O-6 with 1,6-linked α-D-Glcp residue and non-reducing terminal of ß-Glc-1→ through monosaccharide composition, IR, methylation, and NMR analyses. SEM, AFM, and particle size distribution measurements showed that HSP-W had relatively clustered spherical shape with different sizes from nanoscale to micrometer. Additionally, HSP-W significantly induced NO and ROS production as well as the release of IL-1ß, IFN-γ, TNF-α, and IL-6, and upregulated pinocytic and phagocytic capacities of THP-1 cells. Meanwhile, HSP-W treatment markedly enhanced mRNA and protein expressions of these cytokines. HS probably has potential application as an immunostimulatory agent.

Antioxidant efficiency and mechanisms of green tea, rosemary or maté extracts in porcine Longissimus dorsi subjected to iron-induced oxidative stress.

Plant extracts from rosemary (RE), green tea (GTE), and maté (ME) were compared for the protection against iron-induced oxidation in porcine homogenates at total phenolic concentrations from 25 to 250 ppm. Lipid oxidation as indicated by TBARS was in all cases sufficiently suppressed, especially for RE. Hydrophobic RE retarded overall oxidation in the homogenates with an inverted dose-dependent response. Optimum delay of oxygen consumption was found at the lowest concentration applied, similar to protection against thiols and formation of protein radicals as measured by ESR, whereas the high concentration increased oxygen consumption and caused additionally thiol loss possibly due to thiol-quinone interactions, generating protein-phenol complexes. Hydrophilic ME or GTE increased the initial oxygen consumption rate as an indication of prooxidant activities at elevated concentrations. However, they were found to protect myoglobin and protein at those high concentrations with GTE being more efficient, possibly due to better chelation effect.

Interaction of ß-conglycinin with catechin-impact on physical and oxidative stability of safflower oil-in-water emulsion.

The study aimed at improving the antioxidant activity of ß-conglycinin to enhance the oxidative and physical stabilities of safflower oil-in-water emulsion stabilized by ß-conglycinin. Heating promoted binding affinity and antioxidant activity of ß-conglycinin. Catechin and chlorogenic acid showed higher binding affinities towards unheated (or heated) ß-conglycinin than caffeic acid and quercetin. The enhancement efficiencies of the phenolics on the antioxidant activity of unheated (or heated) ß-conglycinin decreased in the order of catechin > quercetin > chlorogenic acid > caffeic acid. Hydrophobic force and hydrogen bonding were the important binding forces for the selected phenolics to ß-conglycinin. The complexation with catechin has no side effect on interfacial behavior and emulsifying property of ß-conglycinin. The use of heated ß-conglycinin-catechin complex as an emulsifier for preparing safflower oil emulsion effectively improved the oxidative and physical stabilities of the emulsion treated with lipoxygenase through inhibition of lipid oxidation, protein carbonyl formation and sulfhydryl loss.

Qualitative analysis on a proposal for key specific medical treatment for aplastic anemia.

OBJECTIVE: By reviewing the medical treatments for aplastic anemia (AA, Suilao Disease), which is the important research interest of Collaborative Group, Key Department of Blood Disease, State Administration of Chinese medicine, the consensus on the diseases have been reached among the different units of the collaborative group. METHODS: Using qualitative analysis, we determined the characteristics, location and pathogenesis of Suilao disease. We discovered the ways of traditional Chinese medical treatment in curing Suilao disease. RESULTS: Acute AA (acute suilao) and chronic AA (chronic suilao) diseases require different treatment. Acute AA requires 3 phrases of treatments, which are "cold", "warm" and "hot". However, chronic AA requires a dialectic treatment, which involves reinforcement of the Shen (Kidney). Suitable Chinese medical treatments for curing Suilao disease were discussed and reached a consensus. CONCLUSION: It is concluded that a summarized therapy approved by many experts could be widely used.

Compound Zhebei granules combined with chemotherapy for the treatment of refractory acute leukemia: a randomized clinical trial.

OBJECTIVE: To observe the effect of compound Zhebei granules (CZBG) with chemotherapy in the treatment of refractory acute leukemia. METHODS: In this multicenter, double-blind, placebo-controlled clinical trial, we used a central (online) randomization system to assign 235 patients to two treatment groups. A total of 118 patients received chemotherapy combined with CZBG (4 g, twice daily) and 117 patients received chemotherapy plus placebo. The clinical efficacy was evaluated at the end of one chemotherapeutic cycle. RESULTS: In the full analysis set, in which deaths due to disease progression were regarded as inefficacy, the rates of complete remission (CR) and partial remission (CR + PR) were 32.35% and 50.00% , respectively, for the chemotherapy combined with CZBG group, and 23.08% and 35.58%, respectively, for the chemotherapy plus placebo group. There was a statistically significant difference between the two groups according to a χ2 test (P < 0.05). In the per protocol analysis set (PPS), the CR (33.67%), CR+PR (52.04%) response rates for the chemotherapy plus CZBG group were significantly different from the response rates of the control group (CR: 24.24% and CR+PR: 37.37%), respectively (P < 0.05). CONCLUSION: CZBG plus chemotherapy can improve the clinical remission rate of refractory acute leukemia after one just one therapeutic cycle.

Busuishengxue ranules mediate their effects upon non-severe aplastic anemia via mitogen-activated protein kinase/extracellular signal-regulated kinase pathway.

OBJECTIVE: To observe the clinical efficacy of Busuishengxue granules on non-severe aplastic anemia (NSAA) and investigate its effect on the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway. METHODS: Sixty NSAA patients were divided equally into two groups. Subjects in the experimental group were treated with Busuishengxue granules, and the control group with Zaizaoshengxue tablets. The treatment course was 6 months and curative efficacy was compared between the two groups as well as with 10 healthy individuals. Flow cytometry (FCM) was used to detect the intracellular concentration of Ca2+ ([Ca2+]i). Western blotting was employed to detect the expression of enzymes in the MAPK/ERK pathway. RESULTS: The efficacy of Busuishengxue granules was significantly better than that of Zaizaoshengxue tablets (P < 0.05). Before treatment, expression of JNK, phospho-ERK 1/2 and p-JNK was higher, and [Ca2+]i higher, than that of the control group (P < 0.05). After treatment with Busuishengxue granules, expression of all enzymes related to signal transduction pathways in the blood cells of NSSA patients were altered to different degrees. CONCLUSION: Busuishengxue granules had a better effect with regard to improving symptom scores, increasing the number of blood leukocytes, and increasing hemoglobin levels than Zaizaosh-engxue tablets, and they differed slightly in terms of increasing the number of platelets.

Effect of busui shengxue granule (see text) on chronic aplastic anemia patients' hematopoietic adhesion molecule VLA-6/CD49f and its ligand laminin.

OBJECTIVE: To observe the effect of Busui Shengxue Granule ((see text) Herbal granule for replenishing marrow to produce blood) on chronic aplastic anemia (CAA) patients' integrin alpha 6 (VLA-6/CD49f) and laminin (Ln). METHODS: Sixty-five patients were divided into experimental group and control group through random number table. There were 34 patients, 17 were male and 17 female, aged 2-67, with a medianage of 30.2 +/- 8.6, in the experimental group, including 17 patients of kidney-yin deficiency and 17 of kidney-yang deficiency, treated by Busui Shengxue Granule. There were 31 patients in the control group, 16 were male and 15 female, aged 4-65, with a medianage of 31.2 +/- 8.0; administered Zaizhang Shengxue Tablet (see text) Herbal tablet for chronic aplastic anemia). Both groups were treated for six months and compared with 10 normal persons after the treatment. Flow cytometry was adopted to detect the change in the expression of VLA-6/CD49f, receptor in mononuclear cells of CAA patients and normal persons. Enzyme-linked immunosorbent assay was applied to detect the expression of peripheral serum Ln. RESULTS: CAA patients' VLA-6/CD49f was in the state of low expression and Ln in the state of high expression. After the treatment, both VLA-6/CD49f and Ln were regulated to some extent and the change in the experimental group was better than that of the control group. Compared with the kidney-yin deficiency patients, those indices of kidney-yang deficiency patients were easier to correct. CONCLUSION: The VLA-6/CD49f and Ln expressions of CAA patients are abnormal. The treatment with Busui Shengxue Granule makes both of them improved.