Radioimmunoassay for neopterin in body fluids and tissues

Specific antibodies against D-erythroneopterin have been prepared in rabbits using a conjugate of D-erythroneopterin to bovine serum albumin (D-erythroneopterinylcaproyl-bovine serum albumin). The antiserum distinguished D-erythroneopterin from other pteridines, i.e., three stereoisomers of neopterin, L-erythrobiopterin, folic acid, xanthopterin, and four other synthetic pteridines. Using this specific antiserum, a radioimmunoassay for D-erythroneopterin has been developed to measure the neopterin concentrations in urine and tissues. The conjugate of D-erythroneopterin with tyramine (NP-Tyra) was synthesized and labeled with 125I as the labeled ligand NP-[125I]tyra for the radioimmunoassay. The minimal detectable amount of neopterin was about 0.1 pmol. The concentration of total neopterin (neopterin, 7,8-dihydroneopterin, quinonoid dihydroneopterin, and tetrahydroneopterin) in the biological samples was obtained by iodine oxidation under acidic conditions prior to the radioimmunoassay, and that of neopterin plus 7,8-dihydroneopterin by oxidation under alkaline conditions. Total neopterin values in human urine obtained by this new radioimmunoassay showed a good agreement with those obtained by high-performance liquid chromatography with fluorescence detection. With rat tissue samples which contained very low concentrations of neopterin as compared to biopterin, biopterin was simultaneously determined by our previously reported radioimmunoassay, and neopterin values were corrected for the cross-reactivity (0.1%). The neopterin concentrations obtained by this method agreed with the values obtained by the radioimmunoassays for neopterin and biopterin after their separation by high-performance liquid chromatography. This very small amount of neopterin, as compared with biopterin, in rat tissues could not be determined by high-performance liquid chromatography-fluorometry alone due to the masking of the neopterin peak by a large biopterin peak.(ABSTRACT TRUNCATED AT 250 WORDS)     

Interaction of anthracycline antibiotics with biopolymers: 9. Comparative study of the interaction kinetics of daunomycin,adriamycin and iremycin with DNA

The interaction kinetics of the three anthracycline antibiotics, daunomycin, adriamycin and iremycin, with calf thymus DNA has been investigated using the temperature-jump technique. Experimental data obtained at high binding ratio have been fitted by a kinetic theory which, for the binding of large ligands to a linear polymer chain, takes into account both nearest-neighbour ligand interaction and the overlap of potential binding sites. The kinetics of such cooperative binding according to a single-step mechanism can be described completely by two independent microscopic parameters, namely one rate constant and a kinetic cooperativity parameter. Both these parameters have been determined for the three anthracyclcine antibiotics, making use of the known equilibrium binding parameters. The association rate constant in the singly contiguous case turns out to be almost the same for all three antibiotics ($\text{7 × 10}{}^6\text{to 8 × 10}{}^6\text{1 mol}{}^{\mathrm{?1}}\text{s}{}^{\mathrm{?1}}$), while the corresponding dissociation rate constant ranges from 3.5 s^?1 for adriamycin to 10 s^?1 for daunomycin and about 35 s^?1 for iremycin. The different equilibrium binding constants thus correspond to different mean attachment times of the antibiotics at the polymer chain, which positively correlate with the inhibitory action of these drugs on in vitro DNA synthesis. Nearest-neighbour interaction in the case of adriamycin-DNA binding kinetics implies that adriamycin molecules dissociate from an isolated binding site nine times more frequently than from a site between two adjacent ligands.     

Immunological characterization of diphtheria toxin recovered from Corynebacterium pseudotuberculosis

Diphtheria toxin (DT) is a potent toxin produced by the so-called diphtheria group which includes Corynebacterium diphtheriae (C. diphtheriae), Corynebacterium ulcerans (C. ulcerans), and Corynebacterium pseudotuberculosis (C. pseudotuberculosis). The present investigation is aimed to study in detail the production of DT by C. pseudotuberculosis. Twenty isolates were obtained from sheep diseased with caseous lymphadenitis (CLA) and twenty-six isolates were obtained from 26 buffaloes diseased with oedematous skin disease (OSD). All isolates were identified by standard microbiological and DT production was assayed serologically by modified Elek test and immunoblotting. All sheep isolates were nitrate negative, failed to hydrolyze starch and could not produce DT, while all buffalo isolates (biotype II) revealed positive results and a specific band of 62 kDa, specific to DT, was resulted in all concentrated cell fractions (CF), but was absent from non-toxigenic biotype I isolates. At the same time, another band of 31 kDa specific to the PLD gene was obtained with all isolates of biotype I and II. Moreover, all isolates showed positive synergistic hemolytic activity and antagonistic hemolysis with β-hemolytic Staphylococci. The obtained results also indicated that C. pseudotuberculosis could be classified into two strains; non-toxigenic biotype I strain, which failed to produce DT as well as being negative to nitrate and starch hydrolysis, and toxigenic biotype II strain, which can reduce nitrate, hydrolyze starch as well as produce DT.     

Enhanced degradation of lignin by the strain fusion of Pseudomonas putida and Gordonia sp

The protoplast fusion technique was applied to construct a more efficient engineering microbial strain to degrade lignin by fusing two strains, Pseudomonas putida and Gordonia sp. At an initial lignin concentration of 900?mg/L, COD, BOD, TOC removal efficiencies increased from 69–76%, 69–72%, and 70–72% by the parent stains to 83%, 83%, and 83% of the fused strain, respectively. IR and HPLC analyses of the treated solution suggested that the fused strains were more capable of breaking the C_α–C_β bonds of the benzene ring in lignin compared to its parent strains, yielding syringyls as the main product. GC–MS analysis was used to identify the release of three-types of lower molecular intermediates: ring-opening, monomer, and dipolymer products. The phenolic hydroxyl group in lignin was oxidized to carbonyls, followed by further degradation to acids and esters. The carboxyl group on the ether linkage that maintains the macromolecular structure of lignin was oxidized to acyls, which further led to depolymerization and the opening of benzene ring.     

四合木（Tetraena mongolica）茎化学成分的GC-MS分析

近年来,特有植物天然成分分析与开发利用已经成为药物化学研究的热点问题之一[1]。四合木( Tetraena mongolica Maxim.)为蒺藜科( Zygophyllaceae)单属植物,仅分布于内蒙古高原和亚洲中部,为中国特有珍稀植物,也为国家二级濒危植物。四合木在防风固沙和维持荒漠生态系统功能方面具有突出的意义;此外,四合木也极易燃烧,在当地被称为“油柴”,因此,四合木有可能成为新的能源植物。目前,对四合木的相关研究主要集中在种群生态学和保护生物学方面[2-4],对四合木化学成分[5-7]及提取物生物活性[8-9]也进行了相关研究。     

使用电子断层技术对IgG1抗体逐个分子的三维重构与结构分析

抗体(antibody)又称免疫球蛋白(immunoglobulin,Ig),是人体免疫反应的重要参与者.了解抗体的结构和结构动态特征,是理解人体免疫作用机理、修复或提高免疫能力、定向设计抗体以治疗各种疾病的基础.本文以人体IgG1抗体为对象,综述了使用透射电子显微学方法研究IgG1抗体结构方向的最新进展.详细介绍了使用逐个分子的电子断层三维重构技术(individual-particle electron tomography,IPET)对抗体进行结构研究的方法,包括样品制备、图像处理和数据分析等.并描述了利用该技术,在研究抗体结合肽分子后的结构形变和通过收集不同构象来研究抗体动态结构特征方面所取得的阶段性成果.最后,对尚待解决的关键问题与该技术未来的发展方向进行了讨论与展望.     

Application of white cellulose acetate sheets on fossil wood investigation

Generally, transparent cellulose acetate sheets as a peel technique material are used in the identification of fossils, whereas white cellulose acetate sheets as a biochemistry technique material are applied in serum protein electrophoresis (SPE). Here we report the application of white cellulose acetate sheets for identifying a polished fossil wood from the Upper Mesozoic of West Liaoning, China. Based on the characters of transverse, radial, and tangential sections, the fossil wood is ascribed to a taxon of Protoglyptostroboxylon sp. Compared with transparent cellulose acetate sheets, white cellulose acetate sheets not only provide the similar information as that of the former, but also are more easily acquired in the Chinese market than the former. Because peel technique supported by white cellulose acetate sheets has the advantages of simple, time-saving, safe, reliable, and practical operation with lower material loss, it is a very good choice for the polished fossil wood investigation in labs, museums, geological parks, and handicraft shops. It is also a convenient approach to training students to learn the anatomic structure of fossil wood.     

Methods to characterize the structure of food powders – a review

Food powders can exist in amorphous, crystalline or mixed structure depending on the order of molecular arrangement in the powder particle matrices. In food production, the structure of powders has a greatly effect on their stability, functionality, and applicability. The undesirable structure of powders can be accidentally formed during production. Therefore, characterization of powder structure as well as quantification of amorphous–crystalline proportions presenting in the powders are essential to control the quality of products during storage and further processing. For these purposes, many analytical techniques with large differences in the degree of selectivity and sensitivity have been developed. In this review, differences in the structure of food powders are described with a focus being placed on applications of amorphous powders. Essentially, applicability of common analytical techniques including X-ray, microscopic, vapor adsorption, thermal, and spectroscopic approaches for quantitative and qualitative structural characterization of food powders is also discussed.     

CO2 emissions from an undrained tropical peatland: Interacting influences of temperature,shading and water table depth

Emission of CO₂ from tropical peatlands is an important component of the global carbon budget. Over days to months, these fluxes are largely controlled by water table depth. However, the diurnal cycle is less well understood, in part, because most measurements have been collected daily at midday. We used an automated chamber system to make hourly measurements of peat surface CO₂ emissions from chambers root‐cut to 30 cm. We then used these data to disentangle the relationship between temperature, water table and heterotrophic respiration (R_het). We made two central observations. First, we found strong diurnal cycles in CO₂ flux and near‐surface peat temperature (<10 cm depth), both peaking at midday. The magnitude of diurnal oscillations was strongly influenced by shading and water table depth, highlighting the limitations of relying on daytime measurements and/or a single correction factor to remove daytime bias in flux measurements. Second, we found mean daily R_het had a strong linear relationship to the depth of the water table, and under flooded conditions, R_het was small and constant. We used this relationship between R_het and water table depth to estimate carbon export from both R_het and dissolved organic carbon over the course of a year based on water table records. R_het dominates annual carbon export, demonstrating the potential for peatland drainage to increase regional CO₂ emissions. Finally, we discuss an apparent incompatibility between hourly and daily average observations of CO₂ flux, water table and temperature: water table and daily average flux data suggest that CO₂ is produced across the entire unsaturated peat profile, whereas temperature and hourly flux data appear to suggest that CO₂ fluxes are controlled by very near surface peat. We explore how temperature‐, moisture‐ and gas transport‐related mechanisms could cause mean CO₂ emissions to increase linearly with water table depth and also have a large diurnal cycle.     

Monitoring Protein Adsorption with Solid-state Nanopores

Solid-state nanopores have been used to perform measurements at the single-molecule level to examine the local structure and flexibility of nucleic acids ^1-6, the unfolding of proteins ⁷, and binding affinity of different ligands ⁸. By coupling these nanopores to the resistive-pulse technique ^9-12, such measurements can be done under a wide variety of conditions and without the need for labeling ³. In the resistive-pulse technique, an ionic salt solution is introduced on both sides of the nanopore. Therefore, ions are driven from one side of the chamber to the other by an applied transmembrane potential, resulting in a steady current. The partitioning of an analyte into the nanopore causes a well-defined deflection in this current, which can be analyzed to extract single-molecule information. Using this technique, the adsorption of single proteins to the nanopore walls can be monitored under a wide range of conditions ¹³. Protein adsorption is growing in importance, because as microfluidic devices shrink in size, the interaction of these systems with single proteins becomes a concern. This protocol describes a rapid assay for protein binding to nitride films, which can readily be extended to other thin films amenable to nanopore drilling, or to functionalized nitride surfaces. A variety of proteins may be explored under a wide range of solutions and denaturing conditions. Additionally, this protocol may be used to explore more basic problems using nanopore spectroscopy.