Mechanism of the Allergenicity Reduction of Ovalbumin by Microwave Pretreatment-Assisted Enzymolysis through Biological Mass Spectrometry.

Ovalbumin (OVA) is a major allergen in hen eggs. Enzymolysis has been demonstrated as an efficient method for reducing OVA allergenicity. This study demonstrates that microwave pretreatment (MP) at 400 W for 20 s assisting bromelain enzymolysis further decreases the allergenicity of OVA, which was attributed to the increase in the degree of hydrolysis and promoted the destruction of IgE-binding epitopes. The results showed that MP could promote OVA unfolding, expose hydrophobic domains, and disrupt tightly packed α-helical structures and disulfide bonds, which increased the degree of hydrolysis by 7.28% and the contents of peptides below 1 kDa from 43.55 to 85.06% in hydrolysates compared with that for untreated OVA. Biological mass spectrometry demonstrated that the number of intact IgE-binding epitope peptides in MP-assisted OVA hydrolysates decreased by 533 compared to that in hydrolysis without MP; consequently, their IgG/IgE binding rates decreased more significantly. Therefore, MP-assisted enzymolysis may provide an alternative method for decreasing the OVA allergenicity.

Ginsenoside Rg1 inhibits high-voltage-activated calcium channel currents in hippocampal neurons of beta-amyloid peptide-exposed rat brain slices.

OBJECTIVE: To examine whether ginsenoside Rg1 (Rg1) inhibits the high-voltage-activated calcium currents (ICa,HVA) via mitogen-activated protein kinase (MAPK) in hippocampal neurons in rat brain slices exposed to beta-amyloid peptide 25-35 (Aß25-35). METHODS: An experimental Alzheimer disease (AD) model was prepared by exposure of rat brain slices to Aß25-35 (10 µmol/L). After treatment with Rg1 (20 µmol/L), the ICa,HVA elicited in hippocampal neurons in these rat brain slices upon depolarization from-40 to 40 mV for 200 ms was recorded by a whole-cell patch clamp to analyze the changes in the peak current density, I-V curve, activation-V curve, and inactivation-V curve. RESULTS: Exposure of rat brain slices to Aß led to a significant increase in ICa,HVA, enhancement of the voltage sensitivity of channel activation, and reduction of the voltage sensitivity of channel inactivation in neurons in the hippocampus of rat brain slices. Rg1 treatment significantly inhibited these changes. These effects of Rg1 could be effectively inhibited by the MAPK inhibitor PD98059. CONCLUSION: Rg1 can inhibit Ica,HVA via MAPK in hippocampal neurons in Aß-exposed rat brain slices.

Optimization of struvite crystallization protocol for pretreating the swine wastewater and its impact on subsequent anaerobic biodegradation of pollutants.

Higher contents of NH(4)(+) and SS in wastewater hamper the anaerobic digestion; necessitating its pretreatment to reduce them. This study reveals optimization of struvite/MAP precipitation protocol followed by anaerobic digestion of pretreated swine wastewater for pollutants removal. Levels of different treatments: stirring speeds, 400 and 160 rpm; pH values, 9.0, 9.5, 10.0, 10.5, 11.0 and 11.5; and P:Mg:N ratios, 1:1:1.2, 1:1:1.7, 1:1:2.2, 1:1:2.7, 1:1:4.0 and 1:1:5.0 were evaluated for MAP crystallization. Among various combinations, protocol comprising of initial 10 min stirring at 400 rpm followed by 160 rpm for 30 min, pH 10.0, and P:Mg:N ratio 1:1:1.2 rendered the best removal efficiency for NH(4)(+), PO(4)(3-), COD, TC and TOC. Subsequent anaerobic biodegradation revealed superiority of MAP supernatant over raw swine wastewater for methane yield and NH(4)(+)-N, PO(4)(3-)-P, COD, TC and TOC removals. It suggests that struvite precipitation as pretreatment to anaerobic digestion is highly effective and advantageous in wastewater treatment.

Effects of plant diversity on nutrient retention and enzyme activities in a full-scale constructed wetland.

This study focused on the relationship between plant diversity (six species richness levels) and nutrient retention and enzyme activities associated with carbon, nitrogen and phosphorus cycling in a full-scale constructed wetland (CW) fed with post-treatment domestic wastewater. Effects of plant species richness on nutrient retention and enzyme activities were assessed using soil chemical and zymological methods, respectively. Retention of NH(4)-N and NO(3)-N in the wetland substrate increased with increasing species richness, while phosphorus retention significantly decreased under the richness level of 16 species per plot. Activities of enzymes such as dehydrogenase, beta-glucosidase, invertase, phenol oxidase, L-arsparaginase, protease and nitrate reductase, while they were affected by plant species richness, were strongly depended on the presence or absence of plants in CW substrate, while activities of enzymes such as CM-cellulase, urease and acid phosphatase were strongly depended on plant species richness. We conclude that plant species richness influenced nutrient retention and enzyme activities in the substrate in our subtropical CW; increase plant species richness in CW will likely improve the efficiency of wastewater treatment.

[The experimental research about the effect of Prunella vulgaris L. on Raji cells growth and expression of apoptosis related protein].

OBJECTIVE: To investigate the anti-lymphoma effect of Prunella vulgaris L. in order to offer exprimental data for the treatment of lymphoma with Prunella vulgaris L. in clinic. METHODS: Effect of Prunella vulgaris L. injection on inhibition ratio of cell growth of Raji cells and IC50 were tested by MTT assay. The growth curve line of Raji cells was drawn also by MTT assay. The cellular morphology was observed by invert microscope, Giemas staining and MTT assay. The expression of apoptosis related protein bcl-2, bax was measured by immunocytochemistry and the quantitative analysis was made with figure analysis system. RESULTS: 1. Prunella vulgaris L. could obviously suppress the cell proliferation of Raji cells in a concentration-dependent manner (r = 0.97). The IC50 was 0.118 mg/ml. 2. After Raji cells was reacted with injection of Prunella vulgaris L. (50 mg/ml) , the morphlogical of apoptosis were observed by invert microscope, Giemsa staining and MTT assay. RESULTS: The results of immunocytochemistry showed that after Raji cells were treated by the injection of Prunella vulgaris L. (50 mg/ml) for 48 hours, the expression of bcl-1 was up-regulated, and the expression of bax was down-regulated. The differences between process group and control group were significant (P < 0.01). CONCLUSION: Prunella vulgaris L. can suppress the proliferation of Raji cells and may be a new anti-lymphoma drug. Inducing the apoptosis of Raji cells maybe one of anti-lymphoma mechanisms.

Study on the effects of Guiqi Oral Liquid in promoting recovery of hematopoiesis in acute irradiation injured mice.

OBJECTIVE: To explore the effects and possible mechanisms of Guiqi Oral Liquid (GQOL) on the recovery of hematopoiesis in acute irradiation injured mice. METHODS: The acute irradiation injured mice were randomly divided into 2 groups: the treated group and the control group, and also a normal control group was set up with 6 mice in it receiving no treatment. After the mice in the former two groups were irradiated by 6.0 Gy (60)Co gamma-ray, every one of them was given 0.4 ml GQOL or saline in equal volume through a gastric tube twice a day for 14 days. On the 4th, 8th and 14th day after irradiation, the bone marrow mononuclear cells (BMMNC) and megakaryocytes in bone marrow tissues of the mice were counted, the proportion of hematopoietic tissues (by area) was measured, and the expression of adhesion molecules, CD44 and CD54, in bone marrow were estimated by immunochemistry. The colony forming unit of spleen (CFU-S) in the mice were counted on the 8th day after irradiation. RESULTS: On the 4th, 8th, 14th day after irradiation, the count of BMMNC and megakaryocyte, and the proportion of hematopoietic tissues in the treated group were higher than those in the control group (P < 0.01 or P < 0.05). CD44 and CD54 expression in the treated group were higher than those in the control group on the 4th and 8th day (P < 0.01), but near normal on the 14th day (P < 0.01). On the 8th day, CFU-S count in the treated group was higher than that in the control group (P < 0.01). CONCLUSION: GQOL can regulate the expression of adhesion molecules, CD44 and CD54, in the bone marrow of the acute irradiation injured mice, which may be one of the mechanisms of GQOL in accelerating the early phase hematopoiesis recovery of mice.

Sensitized effect of beta-cyclodextrin on the fluorescence in the determination of carbaryl.

Based on the significant enhancement of fluorescence intensity of carbaryl in inclusion complex, a spetrofluorimetric method with high sensitivity was developed for the determination of carbaryl in aqueous solution. Under the optimum conditions, the complex had excitation and emission maxima at 278 nm and 332 nm, respectively. The linear range of the method was 7.0 ng/ml-1500 ng/ml with a detection limit of 1.2 ng/ml. The proposed method was successfully used to determine quantitatively of carbaryl in cottonseeds.

[Effect of ligustrazine on expressions of basic fibroblast growth factor and its receptor in bone marrow of mice with acute radiation injury].

OBJECTIVE: To study the expressions of basic fibroblast growth factor (bFGF) and its receptor (bFGFR) in bone marrow of mice with acute radiation injury, and to evaluate the effect of Ligustrazine (Lt) on them. METHODS: Fifty-six Kunming mice of clean grade were randomly divided into 3 groups, the normal group, the control group and the Lt group. Mice in the latter two groups were once homogeneously systemic irradiated with 6.0 Gy of 60Co, with the absorption dose rate of 0.56 Gy/min, then treated with saline (0.2 ml/mice) or Lt (2 mg/mice) respectively, twice a day through gastrogavage for successive 13 days. Mice were sacrificed in batch on the 3rd, 7th and 14th day by cervical dislocation to collect the bilateral femoral bone marrow for preparing bone marrow mono-nuclear cell (BMMNC) suspension. The bFGFR expression on surface of BMMNC was determined by flow cytometry; and the bFGF expression level in one side of femoral bone marrow tissue was detected by immunohistochemistry with SABC-AP assay. RESULTS: The bFGF expression in bone marrow of mice on the 3rd, 7th and 14th day after acute radiation injury all were significantly lower than that of the normal mice (P < 0.05 or P < 0.01). The expressions of bFGF and bFGFR in the Lt group detected were significantly higher than that in the control group detected at the corresponding time points (P < 0.05 or P < 0.01). CONCLUSION: By way of enhancing bFGF expression in bone marrow and bFGFR expression on surface of BMMNC to accelerate the repairing of homopoietic micro-environment in bone marrow might be one of the mechanisms of Lt in promoting hemopoietic function reconstitution after acute radiation injury.