Reactivity of amino acids in the azo coupling reaction: I. Dependence of their reactivity on pH

Azo coupling reactions of N-α-acetylhistidine, N-α-acetyltyrosine, and N-α-acetyllysine with p-methylbenzenediazonium ion were investigated as model reactions to obtain information on the relative reactivity of the histidine, tyrosine, and lysine moieties of protein, separated from structural effects. The azo coupling yields of the amino acids increased as the pH of the reaction medium was increased, indicating that the ractive species are the imidazole anion of histidine, the phenolate anion of tyrosine, and the neutral ε-amino group of lysine. It was calculated, based on percentage yields of the azo products, that the imidazole anion is more reactive than the phenolate anion and the ε-amino group, respectively.     

A new type of 3‐D peripheral ultrastructure in Glaucocystis (Glaucocystales,Glaucophyta) as revealed by ultra‐high voltage electron microscopy

The coccoid glaucophyte genus Glaucocystis is characterized by having a thick cell wall, which has to date prohibited examination of the native ultrastructural features of the protoplast periphery. Recently, however, the three‐dimensional (3‐D) ultrastructure of the protoplast periphery was revealed in two divergent Glaucocystis species, with the world's most powerful ultra‐high voltage electron microscope (UHVEM). The two species exhibit morphological diversity in terms of their 3‐D ultrastructural features. However, these two types do not seem to encompass actual ultrastructural diversity in the genetically diverse genus Glaucocystis. Here, we report a new type of peripheral 3‐D ultrastructure resolved in “G. incrassata” SAG 229‐2 cells by 3‐D modeling based on UHVEM tomography using high‐pressure freezing and freeze‐substitution fixation. The plasma membrane and underlying flattened vesicles in “G. incrassata” SAG 229‐2 exhibited grooves at intervals of 200–600 nm, and the flattened vesicles often overlapped one another at the protoplast periphery. This 3‐D ultrastructure differs from those of the two types previously reported in other species of Glaucocystis. The possibility of classification of Glaucocystis species based on the 3‐D ultrastructure of the protoplast periphery is discussed.     

Evidence for essential lysines in heparin cofactor II

Covalent modification with pyridoxal 5'-phosphate was used to study the function of lysyl residues in heparin cofactor II, a heparin-dependent plasma protease inhibitor. Reduction of the Schiff base with sodium borohydride resulted in modification of 3-4 lysyl residues of heparin cofactor II at high concentrations of pyridoxal 5'-phosphate, one of which was protected in the presence of heparin. The antithrombin activity of modified heparin cofactor II was enhanced compared to the native protein. However, the heparin cofactor activity for thrombin inhibition was reduced significantly or completely eliminated in the modified protease inhibitor depending on the extent of phosphopyridoxylation. In contrast to native heparin cofactor II, the modified protease inhibitor did not bind to a heparin-agarose column. The results suggest that lysyl residues are essential for heparin cofactor activity during thrombin inhibition.     

Chromosomal sites of DNA-membrane attachment in Escherichia coli

Evidence is presented to show that both the chromosomal replication point and the chromosomal origin in Escherichia coli are associated with a structure possessing the sedimentation properties and enzyme sensitivities characteristic of membrane. The data suggest that both newly synthesized DNA strands and template DNA strands are bound at this replication point and that both strands are also bound at the origin.     

冠状病毒HCoV-229E S1蛋白片段的免疫原性分析

目的表达和纯化HCoV-229E的S1蛋白片段（S1 417-547）,分析其诱导的免疫应答。方法将纯化蛋白免疫小鼠,ELISA检测小鼠血清特异性抗体以及血清IL-4和IFN-γ含量,流式细胞术检测小鼠脾脏T淋巴细胞亚群分布,观察其诱导的免疫应答。结果表达蛋白经金属螯合获得纯化,并诱导小鼠产生了高滴度的抗体。脾脏CD4^＋和CD8^＋比例均升高,CD4^＋/CD8^＋比值下降;免疫鼠血清IFN-γ和IL-4水平显著升高。结论成功构建了HCoV-229E S1蛋白的表达载体,并在BL21（DE3）中得到了高效表达,表达蛋白免疫小鼠后诱导了明显的细胞和体液免疫应答。     

Interaction between the urea cycle and the orotate pathway: studies with isolated hepatocytes

Two enzymes catalyze the synthesis of carbamylphosphate (CP) in the liver. One is intramitochondrial and utilizes ammonia to make CP for ureagenesis; the second is cytoplasmic and utilizes glutamine to produce CP for pyrimidine biosynthesis. The extent to which the metabolic independence of the two pathways is abridged by the use of a common precursor was examined with measurements of the incorporation of [14C]NaHCO3 into orotic acid, uridine nucleotides, and urea in isolated hepatocytes. Pyrimidine synthesis was markedly stimulated by physiological concentrations of ammonia, and the stimulation was antagonized by ornithine. At intracellular concentrations of ornithine and levels of ammonia found in the portal circulation, some 90% of pyrimidine synthesis was ammonia-dependent. When the glutamine-dependent activity was released from feedback inhibition with galactosamine, the ammonia-dependent incorporation still accounted for 2/3 of pyrimidine synthesis. These results do not support the widely held view that the cytoplasmic enzyme is the sole source of CP for pyrimidine biosynthesis in the liver. They suggest instead that the bulk of the CP incorporated into hepatic pyrimidines is of mitochondrial origin. However, an experiment with intact animals failed to provide decisive evidence on this interpretation. Pyrimidine biosynthesis was sharply inhibited by the addition of uridine, but ureagenesis was unaffected. When physiological levels of ammonia were provided, the sensitivity of pyrimidine biosynthesis to uridine was lost. Although inhibition of the ammonia-dependent enzyme by pyrimidines has been observed with cell-free preparations, it was not evident in the intact cell. Thus, to the extent that the CP consumed in pyrimidine biosynthesis is of mitochondrial origin, feedback control of the orotate pathway appears to be thwarted.     

Retinal Degeneration Slow (RDS) Glycosylation Plays a Role in Cone Function and in the Regulation of RDS·ROM-1 Protein Complex Formation

The photoreceptor-specific glycoprotein retinal degeneration slow (RDS, also called PRPH2) is necessary for the formation of rod and cone outer segments. Mutations in RDS cause rod and cone-dominant retinal disease, and it is well established that both cell types have different requirements for RDS. However, the molecular mechanisms for this difference remain unclear. Although RDS glycosylation is highly conserved, previous studies have revealed no apparent function for the glycan in rods. In light of the highly conserved nature of RDS glycosylation, we hypothesized that it is important for RDS function in cones and could underlie part of the differential requirement for RDS in the two photoreceptor subtypes. We generated a knockin mouse expressing RDS without the N-glycosylation site (N229S). Normal levels of RDS and the unglycosylated RDS binding partner rod outer segment membrane protein 1 (ROM-1) were found in N229S retinas. However, cone electroretinogram responses were decreased by 40% at 6 months of age. Because cones make up only 3–5% of photoreceptors in the wild-type background, N229S mice were crossed into the nrl^−/− background (in which all rods are converted to cone-like cells) for biochemical analysis. In N229S/nrl^−/− retinas, RDS and ROM-1 levels were decreased by ∼60% each. These data suggest that glycosylation of RDS is required for RDS function or stability in cones, a difference that may be due to extracellular versus intradiscal localization of the RDS glycan in cones versus rods.     

复杂地形下褐飞虱迁飞的数值模拟:个例研究

褐飞虱是影响我国水稻生产的重要迁飞性害虫之一,它的爆发严重影响水稻的生长,并诱发水稻病害,导致水稻减产。研究复杂地形条件下褐飞虱的迁出虫源地、空中迁飞轨迹、降落区,探明复杂地形对褐飞虱迁飞的影响机制,对害虫测报与防治、农业防灾减灾和保障我国粮食安全具有重要的理论意义和实践价值。为了分析覆盖复杂地形的大气动力场、温度场、湿度场对褐飞虱迁入及降落分布的影响,采用WRF-Flexpart耦合轨迹计算模式和GIS空间分析等方法,对2008年9月30日—10月3日发生在广东曲江、肇庆、梅县三站和湖北宜昌、安徽东至、江西吉安三站的褐飞虱迁入和降落过程进行了数值模拟,以揭示复杂地形条件下褐飞虱种群降落和密度分布的时空变化结构。(1)广东曲江、肇庆、梅县三站的后向轨迹模拟结果显示,迁入三站的虫源均来自该站点的西北方向。地形较高且复杂多变时,褐飞虱难以穿越且迁飞距离较短、方向多变。(2)湖北宜昌、安徽东至、江西吉安三站的前向轨迹模拟结果显示,当站点附近的山脉较低且有山谷通道,褐飞虱沿山脉顺风方向迁飞,且迁飞距离较远。当山脉地势较高且没有明显的山谷,则褐飞虱遇到山脉阻挡而转向造成种群滞留。(3)褐飞虱迁飞种群的密度沿山脉走向呈带状分布,山坡较为陡峭、断崖显著时,向远离山体的方向迁飞。若山脉由多个山岭构成,则褐飞虱可从其峡谷穿越,密度分布较为分散。(4)强水平气流有利于褐飞虱的远距离迁飞,下沉气流对褐飞虱降落起着重要的作用,当有强下沉气流且气温较高时,有利于褐飞虱种群的降落,并聚集形成高密度迁入区。(5)垂直方向上,在一定的温度范围内,褐飞虱趋向于在暖层中迁飞。褐飞虱密度沿河谷地带呈带状分布且密度高值区多分布在较温暖的地区。秋季,褐飞虱降落区多分布在相对湿度50%左右的区域。模拟的褐飞虱迁飞轨迹、迁飞方位角和迁飞距离等与实际发生的褐飞虱迁飞路径和降虫区之间偏差较小,该模拟方法较大程度地提高了我国迁飞性害虫的业务预报水平。未来拟提高测报褐飞虱虫情数据的时空分辨率,以为得到较高准度和精度的模拟结果。     

Numerical technique of blood flow through catheterized arteries with overlapping stenosis

This study is concerned with the surgical technique for the injection of a catheter through arteries with overlapping stenosis in the presence of externally applied magnetic field and Hall currents influences. The nature of blood is analyzed mathematically by considering it as a micropolar fluid. The analysis is carried out for an artery with a mild stenosis. The governing equations with the corresponding boundary conditions solved numerically using Crank–Nicolson implicit finite difference scheme. The numerical technique give excellent agreement for axial velocity of the fluid, the circumferential microrotation, the wall shear stress distribution and the contour plots of stream lines. The obtained results show that the value of axial velocity is higher for a Newtonian fluid than that for a micropolar fluid model, the effect of suitable moving magnetic field (Hall currents influences) accelerates the speed of blood, the size of trapped bolus for the stream lines decrease if the spinning movement of the fluid molecules have considerable value regardless of small or large size of the fluid molecules and the flow of fluid is better with increasing the Hall current effect and the size of trapping bolus increase clearly by increasing the maximum height of stenosis where the fluid moves as a bulk.