预热处理金针菇蛋白对猪肉肌原纤维蛋白凝胶特性的影响

本文研究预热处理（preheat treatment,PT）金针菇蛋白（Flammulina filiformis protein,FFP）对肌原纤维蛋白（myofibrillar protein,MP）凝胶特性的影响。试验将FFP在65、75、85和95℃下分别预热处理30、60、90和120min,以蛋白溶解度、总巯基含量和表面疏水性为指标,确定FFP预热处理的最佳温度与时间;将预热处理的FFP（PT-FFP）与MP以不同的比例（0:10、1:9、2:8、3:7、4:6,M/M）混合制备复合凝胶（总蛋白质量浓度均为40mg/mL）,以凝胶强度、持水性、流变学特性与微观结构为指标,考察PT-FFP对MP凝胶特性的影响。结果表明,75℃预热处理60min,FFP的溶解度与表面疏水性最高,总巯基含量最低,故确定该预热处理条件为最佳;将该条件下制备的PT-FFP以不同比例与MP混合制备凝胶,结果发现PT-FFP比FFP更有利于复合凝胶的凝胶强度和持水性的提高（P<0.05）,特别是当PT-FFP与MP混合比例为1:9时,复合凝胶的凝胶强度和持水性达到最高值121.38g和85.25%;流变学结果也表明,PT-FFP能够提高复合凝胶的弹性模量Gʹ和损耗模量G″;电镜观察可以看出PT-FFP与MP复合的凝胶具有更强的连续性,结构更为致密。总之,经过75℃预热处理60min后的FFP和MP以1:9混合时能够改善肌原纤维蛋白凝胶特性和流变学特性。     

三七总皂苷原粉和超微粉的理化性质比较研究

通过对药用植物三七总皂苷原粉和超微粉的粉体粒度、电位、显微结构、红外光谱、溶解速度这几项理化性质的研究,判定两种粉体的优劣,为三七总皂苷超微粉的市场推广提供依据。实验结果表明,三七总皂苷原粉经纳米化处理后成为三七总皂苷超微粉,其化学结构没有发生变化,但显微结构从条状的晶体变为由纳米球紧密排列的不规则形态,其粉体平均粒径也从1 122.4 nm缩小到153.4 nm,完成了从微米到纳米的转变,其水溶液也变为稳定的胶体溶液。运用高效液相色谱法,在模拟人体环境的条件下,发现三七总皂苷超微粉比三七总皂苷原粉早1 min完全溶解。说明三七总皂苷超微粉比三七总皂苷原粉颗粒更小,更易溶于水,更易与人体吸收。     

Temporal variations of membrane foulants in the process of using flat-sheet membrane for simultaneous thickening and digestion of waste activated sludge

Membrane foulants were extracted at different operation time in simultaneous sludge thickening and digestion reactors using flat-sheet membranes. Temporal variations of foulants were analyzed by three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy, gel filtration chromatography (GFC), particle size distribution (PSD) and attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. Results showed that during the first 4 days fouling was mainly assigned to internal membrane foulants (IMFs), and afterwards external membrane foulants (EMFs) increased dramatically. EEM analysis showed that both IMFs and EMFs changed during the operation. Cluster analysis demonstrated that the characteristics of IMFs were relatively similar; however, both quantity and properties of EMFs were changed. GFC analysis showed that EMFs contained more molecules with large molecular weight compared to IMFs. PSD analysis illuminated that particle size of EMFs gradually increased and was larger than that of IMFs. ATR-FTIR analysis indicated that the foulants on membranes consisted of polysaccharides and proteins.     

Effect of ultrasonic treatment on the biochemphysical properties of chitosan

Four chitosans with different molecular weights and degrees of deacetylation degree and 28 chitosans derived from these initial chitosans by ultrasonic degradation have been characterized by gel permeation chromatography (GPC), FT-IR spectroscopy, X-ray diffraction and titrimetric analyses. Antimicrobial activities were investigated against E. coli and S. aureus using an inhibitory rate technique. The results showed that ultrasonic treatment decreased the molecular weight of chitosan, and that chitosan with higher molecular weight and higher DD was more easily degraded. The polydispersity decreased with ultrasonic treatment time, which was in linear relationship with the decrease of molecular weight. Ultrasonic degradation changed the DD of initial chitosan with a lower DD (<90%), but not the DD of the initials chitosan with a higher DD (>90%). The increased crystallinity of ultrasonically treated chitosan indicated that ultrasonic treatment changed the physical structure of chitosan, mainly due to the decrease of molecular weight. Ultrasonic treatment enhanced the antimicrobial activity of chitosan, mainly due to the decrease of molecular weight.     

Evaluation of CO2-based cold sterilization of a model hydrogel

The purpose of the present work is to evaluate a novel CO(2)-based cold sterilization process in terms of both its killing efficiency and its effects on the physical properties of a model hydrogel, poly(acrylic acid-co-acrylamide) potassium salt. Suspensions of Staphylococcus aureus and Escherichia coli were prepared for hydration and inoculation of the gel. The hydrogels were treated with supercritical CO(2) (40 degrees C, 27.6 MPa). The amount of bacteria was quantified before and after treatment. With pure CO(2), complete killing of S. aureus and E. coli was achieved for treatment times as low as 60 min. After treatment with CO(2) plus trace amounts of H(2)O(2) at the same experimental conditions, complete bacteria kill was also achieved. For times less than 30 min, incomplete kill was noted. Several physical properties of the gel were evaluated before and after SC-CO(2) treatment. These were largely unaffected by the CO(2) process. Drying curves showed no significant change between treated (pure CO(2) and CO(2) plus 30% H(2)O(2)) and untreated samples. The average equilibrium swelling ratios were also very similar. No changes in the dry hydrogel particle structure were evident from SEM micrographs.     

青藤碱磷脂复合物纳米结构脂质载体的制备、表征及药动学研究

为制备青藤碱磷脂复合物纳米结构脂质载体,并进行体外和SD大鼠体内评价。实验采用溶剂挥发法制备青藤碱磷脂复合物,乳化超声法制备青藤碱磷脂复合物纳米结构脂质载体。考察其粒径分布、Zeta电位,包封率,载药量及体外释药等基本理化性质。SD大鼠分别灌胃给予青藤碱混悬液和青藤碱磷脂复合物纳米结构脂质载体,比较药动学行为及生物利用度。结果显示,青藤碱磷脂复合物纳米结构脂质载体的平均粒径为201.32±5.05 nm,Zeta电位为-22.2±1.5 mV,包封率为80.31±1.01%,载药量为4.42±0.28%,体外释药具有明显的缓释特征,体外释药模型符合Weibull释药模型,拟合方程为:LnLn(1/1-Mt/M∞)=0.576 6Lnt-1.478 1(r=0.988 8)。体内药动学研究结果表明,磷脂复合物纳米结构脂质载体改变了青藤碱的药动学行为,增强了体内吸收,延长了青藤碱在体内滞留时间,相对生物利用度提高到了1.75倍。因此,青藤碱磷脂复合物纳米结构脂质载体可显著促进青藤碱体内吸收,提高其口服生物利用度。     

Effects of sugars on the cross-linking formation and phase separation of high-pressure induced gel of whey protein from bovine milk

The effects of sugars (xylose, arabinose, fucose, fructose, galactose, glucose, sorbitol, maltose, sucrose, and lactose; 0-20% w/v) on the properties of the pressure-induced gel from a whey protein isolate (20%, 800 MPa, 30 degrees C, 10 min) were studied. All the sugars decreased the hardness, breaking stress and water-holding capacity of the gel at the same concentration of 55.5 mM. Increasing the sugar content changed the microstructure of the gel from a honeycomb-like structure to a stranded structure, while the strand thickness was progressively reduced. These results suggest that sugars decreased the degree of intermolecular S-S bonding of proteins and non-covalent interaction, and restrained the phase separation during gelation under high pressure.     

Enhanced Heat-induced Gelation of β-Lactoglobulin by α-Lactalbumin

Heat-induced gelation in a mixed system of α-lactalbumin (La) and β-lactoglobulin (Lg) was studied to elucidate the gelling properties of whey protein. An Lg concentration of 4% (w/v) was required for the formation of a self-supporting gel following heating at 80°C for 30 min in a 100 mM potassium phosphate buffer (pH 6.8). Solutions of La, even up to a protein concentration of 8% (w/v), did not gel under the same conditions. The addition of 6% La to 2% Lg caused a significant increase in the gel hardness, although each protein did not individually form a gel at these concentrations. By adding La to Lg, firmer gels were formed at a lower heating temperature, compared to that from Lg alone. La and Lg interacted to form a soluble aggregate through a thiol-disulfide interchange reaction during gel formation, and such an interaction was critical in the formation and stabilization of the gel network structure. We conclude that the enhancing effect of La on the gel hardness of Lg was due to the formation of a specific soluble aggregate, and that such an interaction between these proteins contributes to the properties of whey protein gels.     

Inhalable sustained-release formulation of long-acting vasoactive intestinal peptide derivative alleviates acute airway inflammation

The present study was undertaken to develop a respirable sustained-release powder (RP) formulation of long-acting VIP derivative, [Arg(15, 20, 21), Leu(17)]-VIP-GRR (IK312532), using PLGA nanospheres (NS) with the aim of improving the duration of action. NS formulation of IK312532 (IK312532/NS) was prepared by an emulsion solvent diffusion method in oil, and a mixture of the IK312532/NS and erythritol was jet-milled and mixed with lactose carrier to obtain the IK312532/NS-RP. Physicochemical properties were characterized focusing on appearance, particle size, and drug release, and in vivo pharmacological effects were assessed in antigen-sensitized rats. The IK312532/NS with a diameter of 140 nm showed a biphasic release pattern in distilled water with ca. 20% initial burst for 30 min and a sustained slow release up to ca. 55% for 24h. Laser diffraction analysis demonstrated that IK312532/NS-RP had fine dispersibility and suitable particle size for inhalation. In antigen-sensitized rats, insufflated IK312532/NS-RP (10 μg of IK312532/rat) could suppress increases of granulocyte recruitment and myeloperoxidase in pulmonary tissue for up to 24h after antigen challenge, although IK312532-RP at the same dose was less effective with limited duration of action. From these findings, newly prepared IK312532/NS-RP might be of clinical importance in improving duration of action and medication compliance for treatment of airway inflammatory diseases.     

Effect of Particle Size on Compaction of Materials with Different Deformation Mechanisms with and without Lubricants

This work investigates the effect of excipient particle size on compaction properties of brittle, plastic and viscoelastic materials with and without added lubricants. Sieve cuts of Microcrystalline cellulose (MCC), Starch and Dibasic calcium phosphate dihydrate were obtained by sieving, then samples were tested without lubrication or with added lubricant (0.5% Mg stearate mixed for either 5 or 30-min). Compacts were left overnight before testing. It was found that in the absence of lubricant, compact tensile strength (TS) was dependent on particle size only for starch. With Mg stearate, lubricant sensitivity shows a strong dependence on excipient particle size for both starch and MCC, where smaller particles are less affected by lubricant. Dibasic calcium phosphate dihydrate was not sensitive to lubricant even after 30 min mixing. This study highlights that in the absence of lubricant, initial particle size of excipients has no impact on compact strength not only for Dibasic calcium phosphate dihydrate (brittle), but also for MCC (plastic). On the other hand, TS is dependent on particle size both with or without added lubricant for starch (viscoelastic).     

Kinetic properties of bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase from spinach leaves.

A cDNA encoding 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase was isolated from a Spinacia oleracea leaf library and used to express a recombinant enzyme in Escherichia coli and Spodoptera frugiperda cells. The insoluble protein expressed in E. coli was purified and used to raise antibodies. Western blot analysis of a protein extract from spinach leaf showed a single band of 90.8 kDa. Soluble protein was purified to homogeneity from S. frugiperda cells infected with recombinant baculovirus harboring the isolated cDNA. The soluble protein had a molecular mass of 320 kDa, estimated by gel filtration chromatography, and a subunit size of 90.8 kDa. The purified protein had activity of both 6-phosphofructo-2-kinase specific activity 10.4-15.9 nmol min(-1) x mg protein (-1) and fructose-2,6-bisphosphatase (specific activity 1.65-1.75 nmol x mol(-1) mg protein(-1). The 6-phosphofructo-2-kinase activity was activated by inorganic phosphate, and inhibited by 3-carbon phosphorylated metabolites and pyrophosphate. In the presence of phosphate, 3-phosphoglycerate was a mixed inhibitor with respect to both fructose 6-phosphate and ATP. Fructose-2,6-bisphosphatase activity was sensitive to product inhibition; inhibition by inorganic phosphate was uncompetitive, whereas inhibition by fructose 6-phosphate was mixed. These kinetic properties support the view that the level of fructose 2,6-bisphosphate in leaves is determined by the relative concentrations of hexose phosphates, three-carbon phosphate esters and inorganic phosphate in the cytosol through reciprocal modulation of 6-phosphofructo-2-kinase and fructose-2,6-bisphosphatase activities of the bifunctional enzyme.     

拟南芥细胞异三聚体G蛋白的分离纯化

以拟南芥哥伦比亚Columbia(Col-0)野生型悬浮培养细胞为材料,采用超声波破碎、匀浆、离心、40%~60%饱和度硫酸铵分步沉淀、Sephadex G-25脱盐、DEAE-Sepharose Fast Flow离子交换、Sephadex G-200凝胶过滤,最后经过Sepharose CL-6B得到纯化的目的蛋白,蛋白收率为0.097%。纯化的蛋白质经非变性聚丙烯酰胺凝胶电泳鉴定显示为一条带,经Western blotting证实为G蛋白。把经Native-PAGE鉴定的蛋白质的条带回收,进行SDS-PAGE显示有3条带。一条是Gα亚基,其分子量为60kDa左右;另外2条带分子量为45kDa和35kDa,可能是β、γ亚基,初步证实拟南芥中存在异三聚体G蛋白。G蛋白提取方法的建立为在基因突变型拟南芥中G蛋白功能的研究奠定基础。     

湿热预处理对刺芹侧耳蛋白质理化性质、结构和功能特性的影响

以刺芹侧耳Pleurotus eryngii蛋白为研究对象,分别将其在50℃、60℃和70℃下热处理0至30min,探讨了湿热处理对刺芹侧耳蛋白理化性质、结构和功能特性的影响。结果表明,湿热处理后刺芹侧耳蛋白的疏水性先升高后降低（P<0.05）,总巯基含量先降低后升高（P<0.05）,此外,刺芹侧耳蛋白二级结构受到影响,在50℃和60℃下进行湿热处理后,其α螺旋含量降低,而在70℃下则升高,β-折叠含量减少,β-转角和不规则卷曲增加;功能特性方面,刺芹侧耳蛋白在50℃、60℃和70℃湿热处理的20min内,其乳化和发泡性能分别提高（P<0.05）。因此,湿热处理可以增强刺芹侧耳蛋白的功能特性,并改善其在食品加工中的应用。     

Gel chromatography and analytical ultracentrifugation to determine the extent of detergent binding and aggregation, and Stokes radius of membrane proteins using sarcoplasmic reticulum Ca2+-ATPase as an example

For structural studies of integral membrane proteins, including their 3D crystallization, the judicious use of detergent for solubilization and purification is required. Detergent binding by the solubilized protein is an important parameter to determine the hydrodynamic properties in terms of size and aggregational (monomeric/oligo(proto)meric) state of the protein. Detergent binding can be measured by gel filtration chromatography under equilibrium conditions and after separation from mixed micelles of solubilized lipid and detergent. Using sarcoplasmic reticulum Ca(2+)-ATPase as an example, we demonstrate in this protocol complete procedures for measurement of detergent binding using (i) radiolabeled n-dodecyl-beta-D-maltoside (DM) or (ii) from measurements of the increase in refractive index due to the presence of bound detergent on the protein. The latter measurement can also be performed by sedimentation velocity (SV) analysis in the analytical ultracentrifuge which in addition allows determination of the sedimentation coefficient. In combination with estimation of Stokes radius by gel filtration calibration, the molecular mass and asymmetry of the solubilized protein can be calculated. In the proposed protocols, the gel chromatographic procedures require 1 d; SV experiments are performed just after size exclusion. The whole time for these experiments is 24 h. Data analysis of analytical ultracentrifugation requires a couple of days.     

Effects of Extraction Parameters on Particle Size of Titanium Dioxide Nanopowders Prepared by Physical Vapor Deposition Technique

In this work, the effects of different extraction parameters on the particle size of the nanopowders extracted from titanium dioxide thin film samples were studied. These nanopowders were obtained by the conjunctional freezing-assisted ultrasonic extraction method. Titanium dioxide thin films were different in their structures (anatase-only, rutile-only, and mixed phase), and their structural characteristics were determined. Results showed that extraction parameters such as freezing temperature, ultrasonic frequency, and application time are very effective in determining the nanoparticle size, which is very important for many applications and uses of highly pure nanomaterials and nanostructures.

     

超声提取文冠果种仁油及GC-MS成分分析

应用超声提取法提取文冠果种仁油,通过正交实验法考察了种仁粒径、提取时间、物料与溶剂比和超声功率4个因素对提取率的影响,得到了最佳提取工艺条件：粒径为0.5 mm,提取时间为90 min,物料与溶剂比为1：5(w/v),超声功率为175 W,并用GC-MS分析了文冠果种仁油的脂肪酸组分,为进一步开发文冠果种仁油作为生物柴油的原料提供了科学依据。     

基于B超图像评价HIFU治疗剂量对组织损伤程度的影响

基于B超图像分析HIFU治疗中辐照剂量、组织凝固性坏死区域和图像参数之间的关系。通过对高强度聚焦超声辐照新鲜离体组织前后获得的B超图像做数字减影处理,计算图像灰度平均值,同时切片观察并记录生物组织的凝固性坏死区域大小,在此基础上得到大批量数据的统计特性。结果表明：辐照剂量、组织凝固性坏死区域与B超图像灰度平均值在一定范围内成正相关性;当凝固性坏死区域增大到一定程度时,B超图像灰度平均值不再增大,而是呈无规律分布。B超图像灰度可反映组织损伤程度,为实时监控HIFU治疗效果提供依据。     

Enhancement of Ultrasonic Seed Treatment on Yield,Grain Quality Characters,and 2-Acetyl-1-Pyrroline Biosynthesis in Different Fragrant Rice Genotypes

Fragrant rice is popular for the good grain quality and special aroma. The present study conducted a field experiment to investigate the effects of ultrasonic seed treatment on grain yield, quality characters, physiological properties and aroma biosynthesis of different fragrant rice genotypes. The seeds of three fragrant rice genotypes were exposed to 1 min of ultrasonic vibration and then cultivated in paddy field. The results of present study showed that ultrasonic seed treatment increased grain yield of all fragrant rice genotypes but the responses of yield formation to ultrasonic were varied with different genotypes. Compared with control, ultrasonic seed treatment increased grain 2-acetyl-1-pyrroline (2-AP, the key component of fragrant rice aroma) content by 13.40%–44.88%. Ultrasonic seed treatment also reduced the crude protein contents in grains. The head rice rate, rice length, chalky rice rate, and chalkiness degree were influenced by ultrasonic for one or two fragrant rice genotypes. The activities of peroxidase and superoxide dismutase were also enhanced due to ultrasonic seed treatment. In conclusion, ultrasonic seed treatment increased grain, regulated grain aroma and quality, and improved stress resistance of fragrant rice varieties.     

Intracellular L-aspartate-beta-decarboxylase of Pseudomonas sp. and Alcaligenes sp. and its immobilization

Intracellular L-aspartate-beta-decarboxylase of Pseudomonas sp. and Alcaligenes sp. was isolated, purified and characterized. The cells were destroyed by ultrasonic treatment; the enzymes were precipitated by ammonium sulfate fractionation, dialyzed and lyophylized using Biogel P-150. After gel electrophoresis homogeneous enzyme preparations were obtained. The activity of L-aspartate-beta-decarboxylase is rather high--up to 92.1 U/min/mg of protein and is maximal at pH 5.5 and at temperatures of 45-55 degrees C. The Km and Vmax values for the Pseudomonas sp. enzyme are 0.1 M and 0.33 mM/min, respectively: those for the Alcaligenes sp. enzyme are 0.15 M and 1.0 mM/min, respectively. The results of amino acid analysis suggest that the enzymes slightly differ from one another with regard to aspartic and glutamic acid, alanine, valine and isoleucine content. Immobilization of the enzymes on various carriers was performed.     

In vivo inactivation of glycerol dehydrogenase in Klebsiella aerogenes: properties of active and inactivated proteins.

Glycerol:oxidized nicotinamide adenine dinucleotide (NAD+) 2-oxidoreductase (EC 1.1.1.6), an inducible enzyme for anaerobic glycerol catabolism in Klebsiella aerogenes, was purified and found to have a molecular weight of 79,000 by gel electrophoresis. The protein seemed to be enzymatically active either as a dimer of a 40,000-dalton peptide at pH 8.6 or as a tetramer of 160,000 molecular weight at pH 7.0. The enzyme activity was present at high levels in cells growing anaerobically on glycerol, but disappeared with a half-life of about 45 min if molecular oxygen was introduced to the culture. In contrast, no such phenomenon occurred with dihydroxyacetone kinase activity, the second enzyme in the pathway. Immunochemical analysis showed that the inactivation of the oxidoreductase did not involve degradation of the protein. Furthermore, subunits of the active and inactive forms of the enzyme were indistinguishable in size on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and had similar isoelectric points (pH 4.7). Inactivation did, however, alter the gel filtration properties of the enzyme protein and, more importantly, reduced its affinity for the dye Cibacron F3GA and the coenzyme NAD+.