Indirect detection of anti-acetylcholinesterase compounds in microcolumn liquid chromatography using packed bed reactor with immobilized human red blood cell acetylcholinesterase and choline oxidase

The inhibiting compounds were separated by micro-column liquid chromatography in the mobile phase containing the natural substrate acetylcholine. A home-made packed bed microbioreactor system containing immobilized enzyme acetylcholinesterase (ACHE) in human red blood cell membrane and choline oxidase (CHO) from alcaligenes was used for the post-column conversion of acetylcholine to hydrogen peroxide which was detected by an electrochemical detector. The inhibition effect of the solutes caused a decrease in the acetylcholinesterase activity, a decrease in the formation of hydrogen peroxide and also a decrease in the response corresponding to the concentration of the solutes. The rate of the enzyme regeneration was also recorded. The micro-system was compared with a conventional LC system comprising commercially prepared enzyme reactor. The stability of the enzymes is at least 3 weeks at ambient temperature. The limit of detection depends on biological activity of inhibition and for galanthamine was 1 pmol.     

血管紧张素转换酶抑制剂在急性心肌梗塞早期的应用

对２５例急性心肌梗塞（ＡＭＩ）并左心功能不全患者早期应用血管紧张素转换酶抑制剂（ＡＣＥＩ）治疗的对照性临床分析，结果：用小剂量的ＡＣＥＩ治疗对患者心率、血压无显著的影响〔治疗前后心率分别为８９．９０±１０．２２次／分与８９．４２±８．１９次／分，血压分别为１５．３７±３．４６／９．７４±２．３２ｋＰａ（１ｋＰａ＝７．５ｍｍＨｇ）〕和１４．９６±２．６０／９．４６±１．７６ｋＰａ〕；对合并轻、中度泵衰竭患者的疗效显著，但对重度泵衰竭的疗效与对照组比较无显著性差异；治疗组恶性心律失常的发生率较对照组减低１３．３８％，警告性室早的发生率治疗组较对照组减少２６．３９％，但Ⅱ°～Ⅲ°房室传导阻滞的发生率治疗组高于对照组；两组４周病死率比较无显著性差异。作者认为：ＡＣＥＩ治疗ＡＭＩ并轻、中度泵衰竭安全有效；ＡＣＥＩ有一定抗心律失常作用，但应注意传导系统的并发症，使用中应注意各种副作用的发生。     

超声引导胶原酶治疗外侧型腰椎间盘突出症

目的探讨超声引导胶原酶治疗外侧型腰椎间盘突出症的疗效及安全性.方法在相应椎间盘突出的棘突间,旁开1.5～2.5cm处经皮穿刺,采用超声引导定位,穿刺成功后,先进行加压试验,再进行麻醉试验,无腰麻征象后,将胶原酶注入到突出的椎间盘内或周围.结果 231例病人出院时的优良率为92.21%,有效率为99.57%;随访1～5年的优良率为92.65%,有效率为99.58%.近期与远期疗效比较无显著性差异(χ2=0.129,P＞0.05).结论利用超声引导监测、定位,注射胶原酶治疗外侧型腰椎间盘突出症,安全性强、有效率高.     

一株曲霉果糖转移酶的发酵条件及转果糖基活性

对一株曲霉果糖转移酶菌株的产酶培养条件进行了研究。确定了最佳培养基组成：初始蔗糖质量浓度15～18g／dL，氮源为酵母膏，K2HPO4对果糖转移酶的产生具有明显的促进作用，添加0．2g／dLCMC能够使果糖转移酶活力提高到原来的1．3倍，在pH5．5，30℃条件下，果糖转移酶最高酶活力为30．42U／mL。HPLC分析结果表明，转糖基产物为总质量的55．8％。     

Hydroxyethyl starch induced acquired von Willebrand's disease

Hydroxyethyl starch (HES) (Hespan, DPont) is a widely used synthetic volume expander which in standard doses of up to 1.51 in 24 h has no significant effect on coagulation (Munsch et al. 1988). However, the data sheet states that in large volumes HES may alter the coagulation mechanism. We now report a case of HES induced acquired von Willebrand's disease (vWD) in which severe bleeding occurred.     

Homozygosity for the transthyretin-Met30-gene in seven individuals with familial amyloidosis with polyneuropathy detected by restriction enzyme analysis of amplified genomic DNA sequences

Familial amyloidotic polyneuropathy (FAP) with a mutation in position 30 of transthyretin (TTR) (previously called prealbumin) is an autosomal dominant inherited disorder characterized by varying degrees of peripheral neuropathy, nephropathy, gastrointestinal problems, and vitreous amyloid. We have earlier diagnosed homozygosity for the TTR-Met30-gene using Southern analysis in four Swedish individuals. We have found it possible to detect homozygosity for the Met-30 mutation by amplifying discrete regions of the TTR-gene using polymerase chain reaction (PCR), and the amplification products restricted with NsiI analysed by gel electrophoresis. Clinical data on seven homozygous individuals, including three new cases, are presented.     

甘草甜素和甘草酸单铵促进IL-2增强NK活性的研究

本实验采用乳酸脱氢酶释放法研究甘草甜素和甘草酸单铵对人外周血NK细胞活性的影响。结果表明,甘草甜素和甘草酸单铵本身无增强NK细胞活性的作用,但可明显促进IL-2增强NK活性,这种增强作用与甘草甜素和甘草酸单铵的浓度有关。     

Axial tissue diffusion can account for the disparity between current models of hepatic elimination for lipophilic drugs

An assumption of previous models of hepatic elimination is that there is negligible axial diffusion in the liver. We show, by construction of a stochastic model and analysis of published data, that compounds which are readily diffusible and partitioned into hepatocytes may undergo axial tissue diffusion. The compounds most likely to be affected by axial tissue diffusion are the lipophilic drugs for which the cell membranes provide little resistance and which are highly extracted, thereby creating steep concentration gradients along the sinusoid at steady state. This phenomenon greatly modifies the availability of the compound under conditions of altered hepatic blood flow and protein binding. For moderately diffusible compounds, these relationships are similar to those predicted by the simplistic venous-equilibrium model. Hence, the paradoxical ability of the venous-equilibrium model to describe the steady-state kinetics of lipophilic drugs such as lidocaine, meperidine, and propranolol may be finally resolved. The effects of axial tissue diffusion and vascular dispersion on hepatic availability of drugs are compared. Vascular dispersion is of major importance to the availability of poorly diffusible compounds, whereas axial tissue diffusion becomes increasingly dominant for highly diffusive and partitioned substances.This study was supported by the National Health and Medical Research Council of Australia.     

A Series of Serological Tests for the Detection of Antigens and Specific Antibodies in Deep-Seated Candidosis: Experimental Aspects/Eine Gruppe von Antigen- und Antikörpertests zur Serodiagnose von tieflokalisierten Candida-Mykosen: Experimentelle Aspekte

Summary: We describe a series of six serological tests for the diagnosis of deep-seated candidosis. The array comprises two commercial tests (antigen test, Ramco Inc., and antibody test, Roche), as well as four enzyme immunoassays which have been developed in this laboratory: an antigen test for detection of Candida-proteinase, the corresponding assays for monitoring of anti-proteinase antibodies, and two assays for monitoring of IgG and IgM against heterogenous metabolic antigens of C. albicans. The highly sensitive and specific proteinase antigen-test tolerates samples with high concentration of serum proteins. Proteinase antigen was detected in 10 out of 11 normal mice after intravenous infection with C. albicans blastospores. The proteinase antigen peaked between the second and fourth day after infection. A rise in corresponding antibodies was observed in all animals. No proteinase antigen was detected in sera of healthy human individuals; anti-proteinase antibody titers in these sera amounted up to 1:8000. In related ELISAs, using metabolic fungal antigens, titer values of specific IgG and IgM amounted to 5120 and 1280, respectively. The six tests were carried out in an comparative study under diagnostic conditions, the results of which are the subject of a forthcoming communication. Zusammenfassung: Ein Satz von sechs serologischen Tests für die Diagnostik der tiefen Candida-Mykosen wird vorgestellt. Die Gruppe schließt zwei kommerziell vertriebene Testbestecke ein (Latex-Agglutinationstest zum Antigennachweis, Ramco Inc., und Hämagglutinationstest zum Antikörpernachweis, Roche). Vier weitere Enzymimmuntests wurden von uns entwickelt: Ein Antigentest zum Nachweis von sekretorischer Candida-Protease, ein entsprechender Test zum Nachweis von Antikörpem gegen Candida-Protease, und zwei Assays zum Nachweis von IgG-bzw. IgM-Antikörpem gegen heterogene metabolische Antigene von C. albicans. Der empfindliche spezifische Protease-Antigentest toleriert hohe Konzentrationen unspezifischer Serumproteine und kann deshalb auf Serumproben in geringer Verdünnung (z. B. 1:20) angewandt werden. Protease-Antigen war in 200 fach verdünnten Seren von 10 aus 11 intravenös infizierten NWNI-Mäusen nachweisbar. Die höchste Antigen-Konzentration trat zwischen dem 2. und 4. Tag nach Infektion auf; die Serum-Halbwertszeit von gereinigter Protease in der Maus betrug etwa 60 nun. Ein Anstieg korrespondierender Antikörper war in alien infizierten Tieren zu beobachten. Auch im Serum gesunder Probanden waren Antiprotease-Antikörper bis zu einem Titer von 1:8000 nachweisbar; der Protease-Antigentest fiel hingegen immer negativ aus. Die Titer von Antikörpern gegen metabolische Candida-Antigene erreichten in derselben Gruppe von Seren Werte von 1:5120 bzw. 1:1280. Die sechs Tests wurden unter diagnostischen Bedingungen verglichen; Ergebnisse dieser Studie sind Gegenstand einer weiteren Mitteilung.