Optimisation of germination time and temperature on the concentration of bioactive compounds in Brazilian soybean cultivar BRS 133 using response surface methodology

The objective was to optimise the effect of germination time and temperature on the concentration of soluble protein, lunasin, BBI, lectin, saponins and isoflavones in soybean seeds from cultivar BRS 133. Isoflavone and saponin concentrations were analysed by HPLC. Lunasin, Bowman-Birk inhibitor and lectin were analysed by ELISA and western blot. The effects of the variations in germination time and temperature on bioactive compounds were analysed using the response surface methodology (RSM), with a 2² central composite rotational design. Germination of soybean for 42 h at 25 °C resulted in an increase of 61.7% of lunasin, decrease of 58.7% in lectin and 70.0% in lipoxygenase activity. Optimal increases in the concentrations of isoflavone aglycones were observed in combination of 63 h of germination and 30 °C. A significant increase of 32.2% in the concentration of soy saponins was observed in combination of 42 h of germination at 25 °C.     

Water‐soluble protein molecular weight distribution and effects on wheat malt quality during malting

The content and the molecular weight distribution of wheat water‐soluble proteins during malting were determined. Also, the relationships between wheat malt characteristics and water‐soluble protein molecular weight distributions were studied. The water‐soluble protein content was found to increase during malting and the increment was most significant on the first and second days. The molecular weight distribution of the wheat water‐soluble protein was analysed by high‐pressure size exclusion chromatography (HPSEC) and sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS‐PAGE) methods. The HPSEC method showed that the content of all six molecular weight fractions (>24.4, 9.8–24.4, 3.9–9.8, 2.5–3.9, 0.4–2.5 and <0.4 kDa) of the water‐soluble protein increased. Among these, there was a considerable increase in the 0.4–2.5 and 2.5–3.9 kDa fractions. The >24.4, 9.8–24.4 and 3.9–9.8 kDa fractions showed a declining trend during steeping but an increase during the first 2 days of germination. The content of the 9.8–24.4 kDa fraction increased during the process of kilning. Wheat had a high percentage of the >24.4 and 9.8–24.4 kDa fractions, while wheat malt had a high percentage of the 2.5–3.9 and 0.4–2.5 kDa fractions. The molecular weight fractions of the water‐soluble protein had negative correlations with wheat malt viscosity and positive correlations with wheat malt extract. The SDS‐PAGE analysis showed that the band of the 60 kDa water‐soluble protein faded, while the bands of the 56, 44 and 12–15 kDa water‐soluble proteins became darker during malting. Copyright © 2014 The Institute of Brewing & Distilling     

Physicochemical,Pasting, and Functional Properties of Amaranth Seed Flours: Effects of Lipids Removal

The present work was carried out to evaluate physicochemical (composition, hunter color, and sodium dodecyl sulfate–polyacrylamide gel electrophoresis [SDS‐PAGE]), pasting, and functional properties (foaming, emulsification, water, and fat absorption capacity) of amaranth full‐fat flours from 6 lines/cultivars (AFs), and to see the effects of lipid removal/defatting on these properties. Protein, ash, and lipid content of AFs ranged between 12.5% to 15.2%, 3.0% to 3.5%, and 7.1% to 8.0%, respectively. The flours showed a number of bands between 97 and 7 kDa, with main subunits of approximately 58, 37, 33, 31, 23, and 16 kDa in the SDS‐PAGE profiles. The protein content and L* value increased, while b* values decreased following defatting for most of the lines/cultivars. The defatted flours (DAFs) had higher final viscosity and stability (lower breakdown viscosity) as compared to counterpart AFs. The protein profiling of the flours was not affected with the lipid removal/defatting. However, water absorption capacity and foam stability of the flours improved upon defatting. Principal component analysis revealed that pasting temperature was positively related to lipid content, while breakdown viscosity was negatively related to protein content. Foaming properties (capacity and stability) showed negative relationship with lipid content, and positive with protein content, ash content, water, and fat absorption capacity.     

Development of antipeptide enzyme‐linked immunosorbent assay for determination of gelatin in confectionery products

The gelatin sources have become a controversial issue with regard to religious and health concern. Thus, the aims of this study were to develop and evaluate the efficiency of polyclonal antibodies against peptide immunogen of collagen α2 (I) chain for determination of gelatin sources in confectionery products by competitive indirect enzyme‐linked immunosorbent assay (ELISA). Collagen α2 (I) chain protein showed resistance against heat treatment and detectable in certain commercial products when analysed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS‐PAGE). The established ELISA exhibited low cross‐reactivity to fish and chicken gelatin. The IC₅₀ value was 0.39 μg mL^?1, and the limit of detection (IC₁₀) was 0.05 μg mL^?1. There were no false‐positive results from forty‐eight commercially processed products. The present method is useful for determination of gelatin in confectionery products.     

在高浓度尿素的存在下采用制备型电泳体系进行聚丙烯酰胺凝胶电泳分离腐植酸成分

采用制备型电泳体系在高浓度的尿素存在下进行聚丙烯酰胺凝胶电泳分离水溶性腐植酸中的不同成分。在聚丙烯酰胺凝胶电泳中,根据水溶性腐植酸中深色成分分子大小的不同能够将它们彼此分离开来,同时还可以通过酸沉淀的方法将这些深色物质重新回收。水溶性腐植酸中大部分的荧光物质分子尺寸都很小,这使得它们可以溶于酸,并且被酸溶解的荧光物质可以通过吸附在DAX-8树脂上而重新获得。但是,水溶性腐植酸中的非荧光物质则会在电泳过程中丢失。而那些被认为是小分子的荧光物质则会通过氢键的断裂和7 M尿素的疏水作用得到分离。漫反射傅里叶变换红外光谱的测量结果表明腐植酸中不同成分的化学性质是不同的。而实验结果表明在高浓度尿素存在下聚丙烯酰胺凝胶电泳是分离和收集水溶性腐植酸中各组分的一种很有用的方法。     

同位素示踪法测定~(125)I-NGF在小鼠体内的血药浓度变化

本文涉及应用Idogn法对神经生长因子(NGF)进行125I标记并利用同位素示踪与电泳分离相结合的方法测定125I-NGF在小鼠体内的血药浓度与时间变化关系。结果显示,静脉注射125I-NGF在小鼠体内的代谢规律符合二房室开放模型。分布相半衰期为0.13h,消除相半衰期为3.68h,125I-NGF在小鼠体内分布和消除均较快,清除率为0.125L/h/Kg,表观分布容积为0.697L/kg,曲线所围面积为16.01μg.h/L。     

Proteomic profiling of differential display analysis for human oral squamous cell carcinoma: 14-3-3 σ Protein is upregulated in human oral squamous cell carcinoma and dependent on the differentiation level

Oral squamous cell carcinoma (OSCC) has an absolute majority of all oral cancer. We used proteomic technology to analyze the protein expression profile in OSCC tissues and accompanying surrounding normal tissues in four oral locations (buccal mucosa, gingival mucosa, oral floor, and tongue). Ten protein spots were overexpressed more strongly in cancer tissues than normal ones, and were identified as proliferating cell nuclear antigen, 14-3-3 ε, 14-3-3 σ, proteasome subunit α type 5, translationally controlled tumor protein, eukaryotic translation initiation factor 3 subunit, macrophage capping protein, and mitochondrial isocitrate dehydrogenase subunit α. Macrophage capping protein and mitochondrial isocitrate dehydrogenase subunit α had two spots. Especially, we focused on 14-3-3 σ protein, one of the eight identified proteins, and assessed its expression level in four oral locations of OSCC by using differential display methods. The expression level of 14-3-3 σ protein was upregulated in four locations of oral cavity. Eight proteins which we identified in this study may play an important role in OSCC carcinogenesis and progression and could be used as diagnostic biomarkers of OSCC.     

Expression of tropomyosin alpha 4 chain is increased in esophageal squamous cell carcinoma as evidenced by proteomic profiling by two-dimensional electrophoresis and liquid chromatography-mass spectrometry/mass spectrometry

To identify proteins associated with esophageal carcinogenesis, we performed protein profiling of 16 esophageal squamous cell carcinomas (ESCCs) and paired noncancerous tissues by 2-DE and MS/MS. In cancerous tissues, three spots showed significant up-regulation in the amount of protein, while eight spots were significantly down-regulated. The identities of the spots were determined by PMF with LC-MS/MS and were confirmed by immunoblotting. The up-regulated proteins were tropomyosin alpha 4 chain, transgelin, and pyruvate kinase. The down-regulated proteins were serum albumin precursor, isoforms of annexin A1, tropomyosin beta chain, 14-3-3 protein sigma, and isoforms of serotransferrin precursor. In all 16 cases, up-regulation of the tropomyosin alpha 4 chain was confirmed by immunoblotting. Localization of the tropomyosin alpha 4 chain in ESCC cells and adjacent fibroblasts was confirmed by immunohistochemistry.     

两种聚丙烯酰胺凝胶银染方法的比较

比较了两种聚丙烯酰胺凝胶的DNA染色方法。对小麦SSR扩增产物的染色结果显示,改进的Bassam银染法颜色背景浅,灵敏度较高,能够看到副带染色。改进的Sanguinetti方法对条带的鉴别力稍差,但因省时省力,成本较低可用于大规模引物筛选时的染色。     

Effect of non-enzymatic glycosylation of pea albumins on their immunoreactive properties

The aim of this study was to determine the effect of non-enzymatic glycosylation of pea proteins on their immunoreactive properties. Extracted total pea albumins were glycated. No changes were found in molecular weight distribution of total pea albumins before and after glycation using size exclusion chromatography and SDS–PAGE methods. SDS–PAGE GLYCO test stained the glycated proteins and OPA method showed 15% progress in glycation. Glycated and unglycated pea albumins were orally and intraperitoneally administered to Balb/C mice. Serum specific IgG and IgA and sIgA were determined. No difference in serum specific IgG level was found after oral mice immunization with TA and GTA. In the presence of antigen SPL lymphocytes culture showed higher proliferation activity as compared to the culture without the antigen addition. The glycation does not change the immunoreactivity of proteins significantly. During the presented route of immunization with TA and GTA specific tolerance mechanism could be induced.