Adaptive responses in Cypridopsis vidua (Crustacea: Ostracoda) to food and shelter offered by a macrophyte (Chara fragilis)

The ostracod Cypridopsis vidua prefers periphyton growing on Chara fragilis to other feeding substrates. This species actively searches for periphyton, scrapes it off from Chara stems and handles it with a well organized feeding technique. When exposed to different quantities of periphyton on Chara stems, C. vidua selects foraging sites according to their nutritional value. When exposed to low quantities of water from tanks containing fish, C. vidua responds to these weak chemical cues moving from uncovered areas into Chara plants. Experiments with juvenile cyprinids (Abramis brama) as predators and C. vidua as prey clearly showed that the presence of Chara reduces fish foraging success and that C. vidua survival rate increases with increasing density of Chara.     

Characterizing monoclonal antibody structure by carboxyl group footprinting

Structural characterization of proteins and their antigen complexes is essential to the development of new biologic-based medicines. Amino acid-specific covalent labeling (CL) is well suited to probe such structures, especially for cases that are difficult to examine by alternative means due to size, complexity, or instability. We present here a detailed account of carboxyl group labeling (with glycine ethyl ester (GEE) tagging) applied to a glycosylated monoclonal antibody therapeutic (mAb). The experiments were optimized to preserve the structural integrity of the mAb, and experimental conditions were varied and replicated to establish the reproducibility of the technique. Homology-based models were generated and used to compare the solvent accessibility of the labeled residues, which include aspartic acid (D), glutamic acid (E), and the C-terminus (i.e., the target probes), with the experimental data in order to understand the accuracy of the approach. Data from the mAb were compared to reactivity measures of several model peptides to explain observed variations in reactivity. Attenuation of reactivity in otherwise solvent accessible probes is documented as arising from the effects of positive charge or bond formation between adjacent amine and carboxyl groups, the latter accompanied by observed water loss. A comparison of results with previously published data by Deperalta et al using hydroxyl radical footprinting showed that 55% (32/58) of target residues were GEE labeled in this study whereas the previous study reported 21% of the targets were labeled. Although the number of target residues in GEE labeling is fewer, the two approaches provide complementary information. The results highlight advantages of this approach, such as the ease of use at the bench top, the linearity of the dose response plots at high levels of labeling, reproducibility of replicate experiments (<2% variation in modification extent), the similar reactivity of the three target probes, and significant correlation of reactivity and solvent accessible surface area.     

Structural and thermodynamic insights into the assembly of the heteromeric pyridoxal phosphate synthase from Plasmodium falciparum

Pyridoxal 5′-phosphate (PLP) is required as a cofactor by many enzymes. The predominant de novo biosynthetic route is catalyzed by a heteromeric glutamine amidotransferase consisting of the synthase subunit Pdx1 and the glutaminase subunit Pdx2. Previously, Bacillus subtilis PLP synthase was studied by X-ray crystallography and complex assembly had been characterized by isothermal titration calorimetry. The fully assembled PLP synthase complex contains 12 individual Pdx1/Pdx2 glutamine amidotransferase heterodimers. These studies revealed the occurrence of an encounter complex that is tightened in the Michaelis complex when the substrate l-glutamine binds. In this study, we have characterized complex formation of PLP synthase from the malaria-causing human pathogen Plasmodium falciparum using isothermal titration calorimetry. The presence of l-glutamine increases the tightness of the interaction about 30-fold and alters the thermodynamic signature of complex formation. The thermodynamic data are integrated in a 3D homology model of P. falciparum PLP synthase. The negative experimental heat capacity (C_p) describes a protein interface that is dominated by hydrophobic interactions. In the absence of l-glutamine, the experimental C_p is less negative than in its presence, contrasting to the previously characterised bacterial PLP synthase. Thus, while the encounter complexes differ, the Michaelis complexes of plasmodial and bacterial systems have similar characteristics concerning the relative contribution of apolar/polar surface area. In addition, we have verified the role of the N-terminal region of PfPdx1 for complex formation. A “swap mutant” in which the complete αN-helix of plasmodial Pdx1 was exchanged with the corresponding segment from B. subtilis shows cross-binding to B. subtilis Pdx2. The swap mutant also partially elicits glutaminase activity in BsPdx2, demonstrating that formation of the protein complex interface via αN and catalytic activation of the glutaminase are linked processes.     

海金沙草多糖的提取及抗氧化活性(英文)

用响应面优化技术研究了提取时间、固液比、提取温度等对海金沙草粗多糖提取的定量影响,获得了提取工艺的最优工艺参数:提取时间为123.3 min,固液比(s/w)为1∶20.9,提取温度为49.9℃,二阶多项式曲线回归模型预测多糖产量为12.466%,多糖提取验证试验结果(提取率)为12.85±0.18%(n=3),比模型预测稍高。多糖经纯化并进行体外抗氧化活性研究(以维生素C为对照品),结果发现,多糖产物对超氧阴离子(O2–.)和羟基氧自由基(.OH)具有较好的清除作用。     

UV-sensitive P compounds: release mechanism, seasonal fluctuation and inhibitory effects on alkaline phosphatase activity in a shallow Chinese freshwater lake (Donghu Lake)

Filtrable phosphorus compounds in a shallow Chinese freshwater lake (Donghu Lake) were fractionated by Sephadex G-25 gel-filtration chromatography. Some portions of those compounds released soluble reactive phosphorus upon irradiation with low dose ultraviolet light. Catalase and a hydroxyl radical scavenger (mannitol) markedly prevented photosensitive phosphorus release. The observed effects may be explained by the action of oxidizing reagents such as hydroxyl radicals, produced in photochemical reactions between UV irradiation and humic substances in the water. There was a strong seasonality in UV-sensitive P (UVSP) release. Michaels constants (K _m) of total alkaline phosphatase in the lake water showed a direct positive relation to UVSP Plot of K _m against the UVSP/phosphomonoester ratio reveals a strong relationship between the two variables. These results suggest that in some situations UVSP may be a competitive inhibitor of alkaline phosphatase activity in the lake. The competitive inhibition of fractionated UVSP on alkaline phosphatase reagent (Sigma) apparently supports this hypothesis.     

平利短肛棒危害植物种类及其取食量和排粪量的研究

平利短肛棒Baculum pinglienseChenetHe是一种分布较为广泛的新农林害虫 ,可危害 2 0多科、70余种植物的叶片。成虫日平均食鲜栎叶量分别为 :♀ 85 94～ 2 2 2 6 3mg(7 80～ 18 10cm2 )、♂ 19 5 3～ 5 9 5 3mg(1 2 6～4 84cm2 ) ;日平均排粪量 (干重 )分别为 :♀ 36 87～ 70 6 8mg(30 1～ 5 7 7粒 )、♂ 10 36～ 16 6 5mg(4 1 45～ 6 6 5 8粒 )。雌成虫耐饥力比雄成虫强 ,二者耐饥时间长短与温湿度有关     

Study on the polarographic catalytic wave of vitamin P in the presence of persulfate and its application

The polarographic catalytic wave of vitamin P in the presence of persulfate was studied by linear potential scan polarography and cyclic voltammetry. Vitamin P yielded a single reduction wave in acidic aqueous solution, which was ascribed to a 2e(-), 2H(+) reduction of the carbonyl group in the C-4 position. Actually, the carbonyl group C=O first underwent a 1e(-), 1H(+) reduction to form a neutral free radical, and the further 1e(-), 1H(+) reduction of the free radical was simultaneous with its following chemical reactions. When S(2)O(2-)(8) was present, the free radical of vitamin P was oxidized by both S(2)O(2-)(8) and its reduction intermediate, the sulfate radical anion SO(*-)(4), to regenerate the original, which resulted in the production of a polarographic catalytic wave of vitamin P. Based on this catalytic wave, a novel method for the determination of vitamin P was proposed. In 0.02 M tartaric acid-sodium tartrate (pH 3.3) buffer containing 5.0 x 10(-3) M K(2)S(2)O(8), the peak potential of the catalytic wave was -1.42 V (vs SCE) and the peak current was rectilinear to the vitamin P concentration in the range of 8.0 x 10(-9)-1.0 x 10(-6) M (r = 0.9994, n = 13). The catalytic wave of 2.0 x 10(-7) M vitamin P enhanced the polarographic current 70 times compared with the corresponding reduction wave. The detection limit was 2.0 x 10(-9) M, and the relative standard deviation at the 2.0 x 10(-7) M level was 0.7% (n = 15). The proposed method was used for the determination of vitamin P content in the pharmaceutical preparation of tablets and the medicinal plant Sophora japonica L. without previous separation.     

不同致病型的三株IBV与chAPN体外结合能力的比较

目的:比较不同致病型鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)IBV与鸡氨肽酶N体外结合能力的差异。方法:将已构建的p ET-32a-ch APN转入大肠杆菌BL21中进行诱导表达,SDS-PAGE分析表达结果。表达的His-ch APN纯化后进行western-blotting验证,应用酶促反应检测其活性。以不同稀释度的纯化ch APN包被ELISA板,加入经过荧光定量PCR方法定量的含有相同数量病毒粒子的三株不同致病型IBV(呼吸型M41、肾型S-03和腺胃型A2-2),测定三株与ch APN的结合能力。结果:p ET-32a-ch APN在受体菌内成功表达,主要以包涵体的形式存在。经纯化复性的重组His-ch APN,Western blotting分析显示其具有良好的抗原性,酶促反应显示该重组重组His-ch APN与天然的具有良好的生物活性。ELISA结果显示ch APN与三株IBV均能结合,其结合能力为A2-2S-03M41,并表现为剂量依赖性。结论:不同致病型IBV与ch APN的体外结合力有一定差异。     

REGULATION OF FLIGHT CAPACITY AND CONTENTS OF ENERGY SUBSTANCES BY METHOPRENE IN THE MOTHS OF ORIENTAL ARMYWORM,MYTHIMNA SEPARATA*

Abstract The flight capacity and contents of energy substances in the moths of migratory Oriental armyworm, Mythimna separata (Walker)dosed with juvenile hormone analogue methoprene(JHM) were compared with the normal moth through day‐1 to day‐6 after the treatment. The flight capacity of JHM‐treated moths, as determined by a 12‐hours tethered‐flight test, was significantly lower than that of the controls, although it might differ with their age and sex. The moth's flight capacity between the treatments was insignificantly different from the first to 3rd day after the treatment but was significantly different from the 4th to 6th day either in the mean flight duration or in distance. Evaluation of the long fliers at the same stages between the treatments also supported this finding. Flight capacity of the JHM‐treated moths also differed with the sex. The JHM‐treated female did not fly less than the control from day‐1 to day‐3, but did significantly less from day‐4 to day‐6 after the treatment. The flight capacity of the JHM‐treated males, however, was not weaker than that of the control throughout the test period. Comparison of the levels of energy substances in the JHM‐treated adults and the controls showed that JHM may directly acted on metabolism of glyceride, which in turn affected flight capacity. Furthermore, higher levels of glyceride in the treated male adults raised their flight capacity to a stronger levels than the control in most of the days of tethered‐flight test period. Contents of glycogen, however, were insignificantly affected by JHM in both male and female adults. In might therefore be concluded that levels of JH is a critical factor regulationg migratory behavior in the army worm moth. In addition to affect metabolism of glyceride, other possible physiological roles that JH played in determining migrattory behavior are also discussed.     

蝴蝶兰黄花品系光合能力快速测定

以5个蝴蝶兰黄花品系为试材,利用调制叶绿素荧光技术,通过比较其快速光曲线相关参数,以了解不同品系光合能力,尝试建立蝴蝶兰光合能力快速测定技术。结果表明：不同蝴蝶兰黄花品系光合能力存在明显差异,具有较高半饱和光强的品系也具有较高的最大相对电子传递速率,而初始斜率在一定程度上决定了品系最大相对电子传递速率的高低。调制叶绿素荧光技术不仅可用于蝴蝶兰光合能力快速测定,还可应用于蝴蝶兰其他光合生理研究。