表达人胰高血糖素样肽前药工程菌遗传稳定性

通过传代培养的方法,研究高表达重组人胰高血糖素样肽前药(Pro-rhGLPs)的工程菌E.coliBL21(DE3)/pET41a(+)-hGLPs的遗传稳定性,观察菌体和菌落形态,比较在有无选择压力下的质粒稳定性,酶切和测序鉴定重组基因片断的正确性,SDS-PAGE电泳证实重组蛋白质的表达量的稳定性,在C57BL/6小鼠上进行葡萄糖耐量实验检测重组蛋白生物学活性。结果显示:此工程菌连续传代过程中,保持大肠杆菌的典型特征,各代质粒的酶切鉴定和测序正确,重组蛋白表达水平及生物活性也无显著差异。因此,工程菌E.coliBL21(DE3)/pET41a(+)-hGLPs具有良好的遗传稳定性。     

Influence of processing factors on the stability of model mayonnaise with whole egg during long-term storage

Mayonnaise-like oil-in-water emulsions with different stabilities—evaluated from the degree of macroscopic defects, e.g., syneresis—were prepared by different formulations and processing conditions (egg yolk weight, homogenizer speed, and vegetable oil temperature). Emulsions prepared with lower egg yolk content were destabilized for shorter periods. The long-term stability of emulsions was weakly related to initial properties, e.g., oil droplet distribution and protein coverage at the interface. Protein aggregation between oil droplets was observed and would be responsible for the instability of emulsions exhibited by the appearance defects. SDS-PAGE results for adsorbed and unadsorbed proteins at the O/W interface suggested that predominant constituents adsorbed onto the interface were egg white proteins as compared with egg yolk components when the amount of added egg yolk was low. In present condition, egg white proteins adsorbed at the O/W interface could be a bridge of neighboring oil droplets thereby causing flocculation in emulsions.     

Influence of evolution on the stability of ecological communities

In randomly assembled communities, diversity is known to have a destabilizing effect. Evolution may affect this result, but our theoretical knowledge of its role is mostly limited to models of small food webs. In the present article, I introduce evolution in a two-species Lotka-Volterra model in which I vary the interaction type and the cost constraining evolution. Regardless of the cost type, evolution tends to stabilize the dynamics more often in trophic interactions than for mutualism or competition. I then use simulations to study the effect of evolution in larger communities that contain all interaction types. Results suggest that evolution usually stabilizes the dynamics. This stabilizing effect is stronger when evolution affects trophic interactions, but happens for all interaction types. Stabilization decreases with diversity and evolution becomes destabilizing in very diverse communities. This suggests that evolution may not counteract the destabilizing effect of diversity observed in random communities.     

Influence of phosphate anions on the stability of immobilized enzymes. Effect of enzyme nature,immobilization protocol and inactivation conditions

A destabilizing effect at pH 7 of sodium phosphate on several lipases immobilized via interfacial activation is shown in this work. This paper investigates if this destabilizing effect is extended to other inactivation conditions, immobilization protocols or even other immobilized enzymes (ficin, trypsin, β-galactosidase, β-glucosidase, laccase, glucose oxidase and catalase). As lipases, those from Candida antarctica (A and B), Candida rugosa and Rhizomucor miehei have been used. Results confirm the very negative effect of 100 mM sodium phosphate at pH 7.0 for the stability of all studied lipases immobilized on octyl agarose, while using glutaraldehyde-support the effect is smaller (still very significant using CALA) and in some cases the effect disappeared (e.g., using CALB). The change of the pH to 5.0 or 9.0, or the addition of 1 M NaCl reduced the negative effect of the phosphate in some instances (e.g., at pH 5.0, this negative effect is only relevant for CALB). Regarding the other enzymes, only the monomeric β-galactosidase from Aspergillus oryzae is strongly destabilized by the phosphate buffer. This way, the immobilization protocol and the inactivation conditions strongly modulate the negative effect of sodium phosphate on the stability of immobilized lipases, and this effect is not extended to other enzymes.     

不同溶剂和保存条件对尾叶悬钩子花色素苷提取及稳定性的影响

对尾叶悬钩子(Rubus caudifolius Wuzhi)鲜果中花色素苷的提取条件、主要化学组分及pH值、温度、没食子酸和Al^3 对其颜色及稳定性的影响进行了分析探讨。结果表明,尾叶悬钩子花色素苷的主要组分可能为矢车菊素-3-葡萄糖苷;pH值和温度影响该花色素苷的色泽及其稳定性,随着pH值和温度的增加,其分解加剧,且花色素苷的分解均遵循动力学一级反应规律;添加不同浓度的Al^3 ,色素溶液的吸光值有所升高,显示出有一定的增色效应,但Al^3 和没食子酸对贮藏期间花色素苷的稳定性及色泽均无明显效果。     

The effect of substitution patterns on the release rates of opioid peptides DADLE and [Leu(5)]-enkephalin from coumarin prodrug moieties

A coumarin-based prodrug system has been developed in our laboratory for the preparation of esterase-sensitive prodrugs of amines, peptides, and peptidomimetics. The drug release rates from this prodrug system were found to be dependent on the structural features of the drug moiety. The effect of the phenyl ring substitutions on the release kinetics of such prodrugs of model amines was examined recently and it was found that appropriately positioned alkyl substituents on the phenyl ring could help to facilitate the release. Aimed at further understanding the structure-release rate relationship of the coumarin-based cyclic prodrugs, we synthesized and examined a series of substituted coumarinic acid derivatives of opioid peptides, DADLE, and [Leu(5)]-enkephalin.     

Cloning and optimization of induction conditions for mature PsaA (pneumococcal surface adhesin A) expression in Escherichia coli and recombinant protein stability during long-term storage

The gene corresponding to mature PsaA from Streptococcus pneumoniae serotype 14 was cloned into a plasmid with kanamycin resistance and without a purification tag in Escherichia coli to express high levels of the recombinant protein for large-scale production as a potential vaccine candidate or as a carrier for polysaccharide conjugation at Bio-Manguinhos/Fiocruz. The evaluation of induction conditions (IPTG concentration, temperature and time) in E. coli was accomplished by experimental design techniques to enhance the expression level of mature recombinant PsaA (rPsaA). The optimization of induction process conditions led us to perform the recombinant protein induction at 25°C for 16 h, with 0.1mM IPTG in Terrific Broth medium. At these conditions, the level of mature rPsaA expression obtained in E. coli BL21 (DE3) Star by pET28a induction with IPTG was in the range of 0.8 g/L of culture medium, with a 10-fold lower concentration of inducer than usually employed, which contributes to a less expensive process. Mature rPsaA expressed in E. coli BL21 (DE3) Star accounted for approximately 30-35% of the total protein. rPsaA purification by ion exchange allowed the production of high-purity recombinant protein without fusion tags. The results presented in this work confirm that the purified recombinant protein maintains its stability and integrity for long periods of time in various storage conditions (temperatures of 4 or -70°C using different cryoprotectors) and for at least 3 years at 4 or -70°C in PBS. The conformation of the stored protein was confirmed using circular dichroism. Mature rPsaA antigenicity was proven by anti-rPsaA mouse serum recognition through western blot analysis, and no protein degradation was detected after long periods of storage.     

饲用复合酶的浸提条件及稳定性

研究了黑曲霉Aspergillus niger 238发酵曲的最佳浸提条件。结果表明以5倍2％CaCl2溶液,于30℃,120r/min条件下浸提1．5h为佳。通过浓缩得到了浓缩酶液,对浓缩酶液的稳定性进行了初步研究。     

Influence of microbial growth kinetics on steady state multiplicity and stability of a two-step nitrification (SHARON) model

In this paper, the influence of microbial growth kinetics on the number and the stability of steady states for a nitrogen removal process is addressed. A two-step nitrification model is studied, in which the maximum growth rate of ammonium oxidizers is larger than the one of nitrite oxidizers. This model describes the behavior of a SHARON reactor for the treatment of wastewater streams with high ammonium concentrations. Steady states are identified through direct calculation using a canonical state space model representation, for several types of microbial kinetics. The stability of the steady states is assessed and the corresponding phase portraits are analyzed. Practical operation of a SHARON reactor aims at reaching ammonium conversion to nitrite while suppressing further conversion to nitrate. Regions in the input space are identified that result in this desired behavior, with only nitrite formation. It is demonstrated that not only the dilution rate plays a role, as is commonly known, but also the influent ammonium concentration. Besides, the type of microbial (inhibition) kinetics has a nonnegligible influence. While the results indicate that product inhibition does not affect the number of steady states of a (bio)reactor model, it is shown that substrate inhibition clearly yields additional steady states. Particular attention is devoted to the physical interpretation of these phenomena.     

八氯二丙醚(S_2)稳定性的研究

增效剂S2以其增效广谱、效果显著及价廉低毒而受欢迎,但因其稳定性差,目前主要用于蚊香、气雾剂中。为了提高其稳定性,从而更好地开发用于其它剂型或其他领域,通过热贮稳定性实验对其稳定性进行了研究,结果显示高纯度的S2（95％）的稳定性明显优于纯度较低的S2（90％）,加入少量抗氧剂于S2中能显著地降低其氧化分解作用。     

Influence of coupling conditions on activity and operational stability of glucoamylase immobilized on titanium (IV)-activated controlled pore glass

Glucoamylase (EC 3.2.1.3) was coupled to controlled pore glass by using titanium(IV) chloride. The drying conditions used during the activation step were studied, and the highest activity (237 units/g of matrix) of immobilized enzyme was obtained when the support and the titanium(IV) chloride solution were dried at 45°C in vacuo for 16 h. After several washing cycles, the specific activity of the immobilized enzyme was ～13 units/mg of protein irrespective of the washing cycle used. However, this immobilized enzyme preparation was also the least stable ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{= 1}\text{h}$). Investigation of the possibility of the stabilization of the linkage of the enzyme to the support by crosslinking with bifunctional reagents showed that the stabilization of the enzyme ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{=100}\text{h}$) was achievable by treatment with a 5% glutaraldehyde solution at pH 7.0 for 2 h (product activity 67 units/g of matrix, specific activity 4 units/mg of protein); this product also showed no release of protein during use. A higher activity (296 units/g of matrix was achieved by stabilization by treatment with a 5% tannic acid solution at pH 7.0 for 2 h. The combined use of glutaraldehyde and tannic acid was effective in stabilizing the bound enzyme ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{=80 to 120 h}$) with an initial activity of 116 units/g of matrix. When use was made of the same support in presilanized (3-aminopropyltriethoxy silane) form followed by glutaraldehyde coupling a similar initial activity (112 units/g of matrix) was obtained, but the operational stability was much better ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{= 640}\text{h}$.     

Influence of neutral salts on the hydrothermal stability of acid-soluble collagen

The thermal stability of acid-soluble collagens was studied by circular dichroism (CD) spectroscopy. Adult bovine dermal collagen (BDC), rat-tail tendon collagen (RTC), and calf skin collagen (CSC) were compared. Despite some variability in amino acid composition and apparent molecular weight, the CD spectra for helical and unordered collagen structures were essentially the same for all the sources. The melting of these collagens occurs as a two-stage process characterized by a pretransition (T _p) followed by complete denaturation (T _d). The characteristic temperatures vary with the source of the collagen; for mature collagens (BDC, RTC) T _p = 30°C and T _d = 36deg;C, and for CSC T _p = 34°C and T _d = 40°C. Neutral salts, NaCl or KCl, at low concentrations (0.02–0.2 M) appear to bind to the collagens and shift the thermal transitions of these collagens to lower temperatures.     

Influence of physical activity on stability of somatotypes in boys.

The purpose of this study is to examine the stability of somatotypes of boys with different activity levels followed longitudinally for eight years. Thirty-nine boys ated 11 were somatotyped annually (until 18 years) using the Heath-Carter anthropometric rating method. Three sub-groups were formed according to their levels of systematic physical activity designated High, Moderate, and Low Activity. There were no differences in somatochart distributions of the Total Group, but there were considerable individual changes which cancelled each other in group comparisons. All boys changed their somatotype ratings at least once and 67% changed in component dominance. The inter-age correlations for the components were generally of poor prediction value, and the somatotype migratory distances showed considerable variation. The Total Group means differed on three components and this was attributed to differences within the Moderate Activity Group, the only sub-groups to differ. It was concluded that changes occur in somatotypes of adolescent boys, but the influence of physical activity on the changes is inconclusive.     

Influence of loop sequence on relative stability of bimolecular triplex DNA

ＨｏｍｏｐｙｒｉｍｉｄｉｎｅｏｌｉｇｏｎｕｃｌｅｏｔｉｄｅｓｃａｎｂｉｎｄｔｏｔｈｅｍａｊｏｒｇｒｏｏｖｅｏｆｄｕｐｌｅｘＤＮＡａｔｈｏｍｏｐｕｒｉｎｅｈｏｍｏｐｙｒｉｍｉｄｉｎｅｓｅｑｕｅｎｃｅｓ,ｆｏｒｍｉｎｇａｌｏｃａｌｔｒｉｐｌｅｈｅｌｉｘ[1—3].ＴｈｅｒｅａｒｅｓｏｍｅｋｉｎｄｓｏｆｔｒｉｐｌｅｘＤＮＡ,ｕｎｉｍｏｌｅｃｕｌａｒｏｒｂｉｍｏｌｅｃｕｌａｒｏｒｔｅｒｍｏｌｅｃｕｌａｒｔｒｉｐｌｅｘｗｈｉｃｈｃｏｎｓｉｓｔｓｏｆｏｎｅｏｒｔｗｏｏｒｔｈｒｅｅｓｔｒａｎｄｓｏｆＤＮＡ,ｒｅｓｐｅ…     

长期免耕不同秸秆覆盖量对玉米产量及其稳定性的影响

阐明长期免耕不同秸秆覆盖量下玉米产量变化趋势及其稳定性差异,可为建立和评价长期保护性耕作模式、促进粮食持续生产提供理论支撑.本研究基于我国东北黑土区长期保护性耕作定位试验(始于2007年),以传统垄作(RT)为对照,分析了免耕无秸秆覆盖(NT0)、免耕33％秸秆覆盖(NT33)、免耕67％秸秆覆盖(NT67)和免耕10...     

Developmental stability in flowers of Clarkia tembloriensis (Onagraceae)

Developmental instability of floral traits is examined in four populations of Clarkia tembloriensis (Onagraceae) with different natural outcrossing rates. Developmental instability is estimated using fluctuating asymmetry (FA) and within plant variance. The results are coupled with those from a previous study of leaf traits. In the first experiment, flowers were collected from the same growth chamber-grown plants that had been previously used to estimate leaf developmental stability in two C. tembloriensis populations. These populations differed in FA for only one floral trait, long filament length. After adjusting for organ size differences, we found floral FA values were about half those of leaves. These are the first quantitative data indicating that flowers are more developmentally stable than leaves. In a second experiment, greenhouse grown plants from two other C. tembloriensis populations (one highly outcrossing and one predominantly self-pollinating) did not differ significantly in floral FA or in within-plant variance of floral traits, though earlier studies of the same populations revealed significant differences in FA of leaf traits. In both experiments, FA values of different floral traits were uncorrelated. We attribute the lack of significant differences in floral stability between populations to the greater canalization of floral organs and to the magnification of measurement error that occurs when calculating FA. We also found that the shorter styles of selfers are the greatest difference in flower form between predominantly self-pollinating and predominantly outcrossing populations of C. tembloriensis.     

Quantifying change in pelagic plankton network stability and topology based on empirical long-term data

Over the last 34 years, Lake Müggelsee has experienced concurrent warming and nutrient reduction. While the effects of environmental change on single taxonomic or physical–chemical variables have been relatively well researched in isolation, understanding how environmental change propagates through the ecological network remains a major challenge. Capitalizing on the long-term monitoring program of the German Long-Term Ecosystem Research Network site Lake Müggelsee (1979-ongoing), we identified three time periods (1979–1995; 1996–2005; 2006–2013) which differed significantly in phytoplankton biomass and relative plankton community composition. Using multivariate first order autoregressive (MAR1) modeling on 13 pelagic plankton groups and four abiotic variables, we quantified interaction networks and indicators of stability and centrality for each period. Our results suggested that the Müggelsee network was bottom-up regulated in all periods and that stability increased over time. Moreover, in all three networks, non-trophic and indirect interactions appeared to be as commonly present as trophic and direct interactions. Using network centrality measures of betweenness and closeness, we identified keystone plankton groups and groups particularly responsive to environmental change based on variation in centrality ranks over time. Given a more comprehensive understanding of the interaction network at hand, MAR1 model-derived stability and centrality measures may potentially be used as integrated ecological indicators to monitor changes in stability of lake ecosystems and to identify particularly vulnerable components of the network.     

Influence of environmental (culturing) conditions on the lectin-binding capacity of Tetrahymena

Extreme intra-assay variations and/or contradictory results frequently encountered in lectin binding studies on Tetrahymena cultures prompted us to investigate the environmental factors affecting the physical or chemical properties of the cell membrane for influence on the lectin-binding capacity thereof. It was found that changes in ambient temperature, osmolarity and illumination, as well as temporary starvation, modified the lectin-binding capacity of Tetrahymena to an appreciable degree. It follows that the comparability of the experimental results presupposes the standardization of the above environmental parameters in Tetrahymena cultures used for lectin-binding studies.     

Sufficient and necessary conditions for Lyapunov stability of genetic networks with SUM regulatory logic

In this paper, a nonlinear model for genetic regulator networks (GRNs) with SUM regulatory logic is presented. Four sufficient and necessary conditions of global asymptotical stability and global exponential stability for the equilibrium point of the GRNs are proposed, respectively. Specifically, three weak sufficient conditions and corresponding corollaries are derived by using comparing theorem and Dini derivative method. Then, a famous GRN model is used as the example to illustrate the effectiveness of our theoretical results. Comparing to the results in the previous literature, some novel ideas, study methods and interesting results are explored.