Resolution of lymphocyte subpopulations derived from rat spleen by polyamine-graft-PHEMA copolymer

The retention behavior of lymphocyte subpopulations, B cell, T cell and null cell, derived from rat spleen to polyamine-graft-poly(2-hydroxyethyl methacrylate) copolymer (HA) surface was investigated, focusing on the conformational transition of the polyamine side chain as well as the protonation of amino groups in the polyamine grafts. Furthermore, the availability of HA was discussed as a column adsorbent for separation of lymphocyte subpopulations derived from spleen. The conformational transition of polyamine grafts significantly influenced the mode of retention of lymphocyte subpopulations. When polyamine grafts existed in an aggregated conformation (protonatin degree α < 0.5), the retention of lymphocyte subpopulations was decreased in the order B cell> null cell> T cell. On the other hand, when polyamine existed in an extended conformation into the aqueous interior from the matrix interface (α > 0.5), T cell retention became greater than null cell retention, resulting in a decreased B cell> T cell> null cell order. These results indicate that the differential retention of spleen lymphocyte subpopulations is attributed to their differential responses to the change in matrix interface accompanied by the protonation of amino groups. Furthermore, spleen lymphocytes were compared with lymph node lymphocytes in terms of resolution efficacy by an HA copolymer column.     

含离子液体［bmim］Cl的反应介质中马肝醇脱氢酶的催化特性

 研究了马肝醇脱氢酶(HLADH)在含离子液体1-丁基-3-甲基咪唑氯酸盐(［bmim］Cl)的反应介质中的催化特性. 以乙醇为底物时,该酶在［bmim］Cl含量≤0.15 g/ml的体系中的活力高于在不含离子液体的体系中的活力; 离子液体浓度过高(＞0.15 g/ml)对酶活性有明显的抑制作用. 反应温度和pH对含离子液体的反应介质中酶活力的影响规律与不含离子液体时的规律相似. 与不含离子液体的反应介质相比, HLADH在含0.05 g/ml ［bmim］Cl的体系中催化乙醇氧化的活化能下降,酶反应的Vmax和Km均升高. 反应体系中低浓度(≤0.1 g/ml)的离子液体能提高酶的热稳定性,但高浓度(＞0.1 g/ml)的离子液体可降低酶的热稳定性. 紫外二阶导数光谱显示,在含不同浓度离子液体的反应介质中酶分子构象的变化有较大的差异.     

Solid phase extraction and analysis of horse urine for aminocaproic acid

Summary A rapid and efficient procedure is described for extraction and determination of aminocaproic acid in horse urine. Urine was extracted by passing through a bonded silica column (Bond-Elut). The adsorbed drug was washed free of endogenous materials before being eluted. The extract was then examined by thin-layer chromatography and HPLC. The purity of the extract was determined by gas chromatography-mass spectrometry.     

Conjugates of Iron-Transporting N-Hydroxylactams with Ciprofloxacin

Screening of a library of novel N-hydroxylactams amenable by the Castagnoli-Cushman reaction identified four lead compounds that facilitated ⁵⁵Fe transport into P. aeruginosa cells (one of these synthetic siderophores was found to be as efficient at promoting iron uptake as the natural siderophores pyoverdine, pyochelin or enterobactin). Conjugates of the four lead siderophores with ciprofloxacin were tested for antibacterial activity against P. aeruginosa POA1 (wild type) and the ∆pvdF∆pchA mutant strain. The antibacterial activity was found to be pronounced against the ∆pvdF∆pchA mutant strain grown in CAA medium but not for the POA1 strain. This may be indicative of these compounds being ‘Trojan horse’ antibiotics. Further scrutiny of the mechanism of the antibacterial action of the newly developed conjugates is warranted.     

健脾八珍颗粒的研制

为筛选出最佳的健脾八珍颗粒处方,以其成型性、溶解性为筛选指标,筛选出处方中加入辅料的品种及配比。结果表明,健脾八珍颗粒的原粉中加入2倍量的糊精和25%的蔗糖,并以2%的CMC-Na作为助悬剂、15%的自制变性淀粉胶(SWJ)为黏合剂为最佳处方。所得产品溶解性、成型性能均较好。     

Spleen and thymus metabolomics strategy to explore the immunoregulatory mechanism of total withanolides from the leaves of Datura metel L. on imiquimod-induced psoriatic skin dermatitis in mice

Our previous work demonstrated that total withanolides of Datura metel L. leaves (TWD) exhibited excellent therapeutic effects on psoriasis. However, current knowledge of its mechanisms is incomplete. In this study, integrated spleen and thymus untargeted metabolomics were used to analyze the changes in endogenous metabolites underlying the immunosuppressive activity of TWD on psoriasis animal models induced by imiquimod. The results suggested that TWD treatment markedly attenuated imiquimod-induced psoriasis and showed significant immunosuppressive activity as evidenced by decreased elevation index of spleen and thymus. Meanwhile, TWD significantly reversed the elevation of immunoregulatory factors, including IL-10, IL-17, IL-22 and IL-23. Multivariate trajectory analysis revealed that TWD treatment could restore the psoriasis-disturbed spleen and thymus metabolite profiles towards the normal metabolic status. A total of 25 and 27 metabolites associated with the immunomodulatory effects for which levels changed markedly upon treatment have been identified in spleen and thymus, respectively. These differential metabolites were mainly involved in amino acid metabolism, nucleotide metabolism, fatty acid metabolism and lipid metabolism. Our investigation provided a holistic view of TWD for intervention in psoriasis through immunoregulation and provided further scientific information in vivo about a clinical value of TWD for psoriasis.     

Modification of the Fractionated γ-Radiation-Induced Alteration in the Na+/K+-ATPase Activity of Radiosensitive Tissues by Vitamin A

The effect of pre or post-administration of vitamin A in ameliorating the radiation-induced alteration in intestinal and spleen Na⁺/K⁺-ATPase activity of rats was determined. Rats subjected to multifractionated γ-radiation (20 × 0.5 Gy) over the whole-body showed a significant reduction in Na⁺/K⁺-ATPase activity being more obvious in spleen than in intestine. Administration of vitamin A at a daily i. p. dose of 15,000 IU/kg bd. wt. for 7 days prior to fractionated γ-irradiation and thereafter twice weekly throughout the exposure period (as pretreatment regimen) or following the end of irradiation for 7 days (as posttreament regimen) afforded a significant protection from radiation-induced inhibition of spleen and intestinal Na⁺/K⁺-ATPase activity. Even though, pretreatment regimen is more efficient against radiation injury to this enzyme than posttreatment, so that its activity was nearly normalized in the first regimen. The regeneration of normal enzyme activity in the tissues of the vitamin A-pretreated group suggests that the intestinal and spleen epithelia in presence of vitamin A are rendered relatively radioresistant.     

Horse radish peroxidase immobilized polyaniline for hydrogen peroxide sensor

Horse radish peroxidase (HRP) has been electrochemically entrapped into perchlorate (ClO) doped polyaniline (PANI) film deposited onto indium‐tin‐oxide (ITO) coated glass plate. This HRP‐PANI‐ClO/ITO bioelectrode characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), UV‐Visible spectroscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) techniques has been utilized for estimation of hydrogen peroxide (H₂O₂). This H₂O₂ sensor exhibits response time of 5 s, linearity from 3 to 136 mM, sensitivity as 0.5638 µA mM^?1 cm^?2 with linear regression of 0.985. The value of the Michaelis–Menten constant (K_m) has been obtained as 1.984 mM. Copyright © 2009 John Wiley & Sons, Ltd.     

Use of a porous silicon–gold plasmonic nanostructure to enhance serum peptide signals in MALDI-TOF analysis

Small peptides in serum are potential biomarkers for the diagnosis of cancer and other diseases. The identification of peptide biomarkers in human plasma/serum has become an area of high interest in medical research. However, the direct analysis of peptides in serum samples using mass spectrometry is challenging due to the low concentration of peptides and the high abundance of high-molecular-weight proteins in serum, the latter of which causes severe signal suppression. Herein, we reported that porous semiconductor-noble metal hybrid nanostructures can both eliminate the interference from large proteins in serum samples and significantly enhance the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) yields of peptides captured on the nanostructure. Serum peptide fingerprints with high fidelity can be acquired rapidly, and successful discrimination of colorectal cancer patients based on peptide fingerprints is demonstrated.     

Chromatographic Detection of Trimetoquinol (Inolin®) and its Major Urinary Metabolites in the Horse: A Preliminary Report

Trimetoquinol (TMQ) (1-(3,4,5-trimethoxybenzyl)-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline, m.w. 345) is the prototype tetrahydroisoquinoline pharmaceutical. TMQ is marketed as a bronchodilator in human medicine; in horse racing, TMQ is listed as an Association of Racing Commissioners International (ARCI) class 3 foreign substance. As such, TMQ is considered to have the potential to affect racing performance in horses, and a validated qualitative confirmatory method is required to regulate its use in racing. We selected 8 g kg^–1 of TMQ IV as a safe and effective dose for studies on its metabolism and analytical detection in horses. We developed a solid phase extraction method for recovery of TMQ and its metabolites from equine urine, identified suitable high performance liquid chromatographic conditions for these substances and our internal standard, papaverine, and developed a highly sensitive ESI(+)-LC-MS-MS method (estimated LOD, 100 pg mL^–1) for TMQ and its major metabolites in equine urine. Multiple Reaction Monitoring (MRM) analysis of unhydrolyzed post-administration urine showed small amounts of unchanged TMQ, along with glucuronide, methylated, and sulfated metabolites, with glucuronide metabolites predominating. Following glucuronidase hydrolysis, recovered parent TMQ peaked at relatively high concentrations (>300 ng mL^–1) within 1 h of administration and thereafter declined. The methylated metabolites of TMQ peaked later and at comparable total concentrations, and thereafter declined more slowly. These data suggest that glucuronide hydrolysis of post-administration urine samples will allow recovery of readily identifiable quantities of parent TMQ. These findings, combined with the highly sensitive LC-MS-MS detection of parent TMQ described herein suggest that glucuronide hydrolysis of post-administration urine, followed by LC-MS-MS or other analysis, will allow effective regulatory control of this agent in racing horses.Published as # 351 from the Equine Pharmacology, Therapeutics and Toxicology Program at the Maxwell H. Gluck Equine Research Center and Department of Veterinary Science, University of Kentucky. Published as Kentucky Agricultural Experiment Station Article # 04-14-048 with the approval of the Dean and Director, College of Agriculture and the Kentucky Agricultural Experimental Station.Revised: 8 June and 12 July 2004