Mapping of murine YACs containing the genesCea2 andCea4 after B1-PCR amplification and FISH-analysis

PCR with primers specific for the murine B1 consensus sequence allows amplification of DNA from murine sources. We have used B1-PCR for amplifying yeast artificial chromosome (YAC) DNA which can be used to localize single YACs by fluorescencein situ hybridization. The genes for the pregnancy-specific glycoproteins Cea2 and Cea4, both belonging to the large carcinoembryonic antigen gene family, were localized by chromosomalin situ suppression hybridization of three YAC clones to murine chromosome 7A2-A3. This was facilitated by the use of the mouse lymphoma cell line WMP/WMP which contains nine pairs of Robertsonian fusion chromosomes.     

Congenital muscular dystrophy and severe central nervous system atrophy in two siblings

Severe degenerative features of the nervous system of a hitherto unknown kind, associated with a neuromuscular disorder with histopathological features of congenital muscular dystrophy, are reported in two female siblings. The clinical profile was characterized by generalized hypotonia followed by spastic tetraplegia, contractures, polyneuropathy, lack of cognitive development and progressive microcephaly. There was no involvement of the eyes. Neuropathological examination of the brain of one sibling, who died at the age of 30 months, revealed subtotal loss of neurons in the cerebral and cerebellar cortex and in the ventral pons, and secondary loss of myelin in the cerebral and cerebellar subcortical white matter. Sural nerve biopsy in the other sibling, who had a similar neurological affection, showed a lack of large myelinated fibers.This investigation is part of the research program Disorders of the Neuromuscular System of the University of Nijmegen     

API0134对血小板α颗粒膜蛋白和血小板聚集的影响

２０例急性心肌梗死患者在溶栓后随机分为穿心莲有效成分（ＡＰＩ０１３４,ＡＰＩ）组与对照组,治疗１５ｄ,均同时给予阿司匹林等药物。检测溶栓前后血浆α颗粒膜蛋白（ＧＭＰ－１４０）浓度和ＡＤＰ诱导的血小板聚集反应。结果表明,ＡＰＩ组溶栓后７２ｈ血浆ＧＭＰ－１４０浓度轻度增加,对照组显著性增加（Ｐ＜０．０１）。ＡＰＩ组ＧＭＰ－１４０在溶栓后７２ｈ和１５ｄ均低于对照组,差异有显著意义（均为Ｐ＜０．０５）。ＡＰＩ组１ｍｉｎ和５ｍｉｎ血小板聚集率均低于对照组,差异均有显著意义（７２ｈ,Ｐ＜０．０５;１５ｄ,Ｐ＜０．０１）。研究表明,ＡＰＩ与阿司匹林联合应用可抑制溶栓后血小板的活化,优于阿司匹林单独应用,因此ＡＰＩ有可能用于临床预防溶栓后的再闭塞。     

Black-gold: a simple, high-resolution histochemical label for normal and pathological myelin in brain tissue sections

A novel haloaurophosphate complex called Black-Gold has been synthesized and applied to localize myelin within the central nervous system. The technique is tailored to studies using formalin fixed non-solvent processed tissue. The technique stains large myelinated tracts dark red-brown, while the individual myelinated axons appear black. This study demonstrates how this novel tracer can be used to localize both normal and pathological myelin. Specific myelin changes associated with exposure to diverse neurotoxicants including kainic acid, domoic acid, 3-nitropropionic acid, Fluoro-Gold and isoniazid are demonstrated and characterized. This study also demonstrates how Black-Gold can be combined with other histochemical markers including Nissl stains, retrogradely transported fluorescent tracers and fluorescent markers of neuronal degeneration. Advantages associated with the Black-Gold technique include high resolution, high contrast, short histochemical processing time, and consistent reproducibility.     

Binding ofβ-adrenoceptor blocking drugs to human serum albumin,to α1-acid glycoprotein and to human serum

Summary The binding of 8 -adrenergic blocking drugs to human serum albumin, to ₁-acid glycoprotein and to serum from normal volunteers and from patients with rheumatoid arthritis was studied. Protein binding was determined in vitro using equilibrium dialysis of labelled drug at 25° C. Oxprenolol and propranolol were highly bound to serum, alprenolol, pindolol and timolol to a lesser degree, and atenolol, metoprolol and sotalol were negligibly bound. For the five compounds which were appreciably bound, the mean binding was significantly higher in serum from patients with rheumatoid arthritis than in serum from normal volunteers. For those drugs, binding to ₁-acid glycoprotein was higher than to human serum albumin, and binding to a mixture of both proteins approached that to serum from healthy volunteers. For each of these drugs there was a strong correlation between the serum ₁-glycoprotein concentration and the percentage binding.     

Binding of aprindine and moxaprindine to human serum, α1-acid glycoprotein and serum of healthy and diseased humans

Summary A comparison was made between the binding of the anti-arrhythmic agents aprindine and moxaprindine to human serum, to human serum albumin (HSA), to ₁-acid glycoprotein (₁-AGP) and to a mixture of HSA and ₁-AGP. In serum from healthy volunteers (n=4) the binding of aprindine-HCl 5 µg/ml (13.8 µM) was 93.8% (SD±1.0), and that of moxaprindine-HCl 5 µg/ml (12.8 µM) was 94.1% (SD±1.1). Their binding to the mixture of ₁-AGP and albumin approximated their binding to serum. For ₁-AGP, the binding was similar for both compounds, whereas for HSA the binding of aprindine was more pronounced than that of moxaprindine: for both products the affinity coefficient for binding to ₁-AGP was about 100 times greater than that for binding to albumin. In serum from rheumatoid patients and from patients with renal failure a small but significant increase in binding of aprindine and moxaprindine was observed, approximately 1%. Increased and decreased binding was seen in serum from cirrhotic patients; for example, for aprindine the range in cirrhosis was 96.7%–79.8%, and the range in controls was 95.0%–92.4%. Free drug fraction and ₁-AGP concentration were inversely correlated. The results show that ₁-AGP plays an important role in the binding of aprindine and moxaprindine, and that alteration in the binding of the two compounds in disease states to a large extent can be explained by changes in serum ₁-AGP concentration.     

Serum concentration and protein binding of thioridazine and its metabolites in patients with chronic alcoholism

Summary The blood chemistry and clinical pharmacokinetics of thioridazine and its metabolites, side-chain sulphoxide, side-chain sulphone and ring sulphoxide, were studied in 31 alcoholics and were compared with values in 17 thioridazine-treated controls without alcoholism. Pathological blood chemistry values, including abnormal liver function and protein concentrations, were common among the alcoholics. In relation to dosage, the majority had a low serum concentration of thioridazine and at a given concentration of thioridazine they had high serum concentrations of its metabolites. Positive intercorrelations were found between pathological liver function tests, prolonged serum half-life and increased serum concentration of thioridazine. The free fractions of thioridazine, side-chain sulphoxide and ring sulphoxide were significantly higher and those of the side-chain sulphone lower in the alcoholics than in the controls. The free fractions of side-chain and ring sulphoxide were significantly increased in patients with a low concentration of ₁-acid glycoprotein.     

Hereditary neuropathy with liability to pressure palsies

Summary Clinical, neurophysiological and pathological investigations were carried out in 11 affected members of 2 families with hereditary neuropathy with liability to pressure palsies (HNPP). The observations were related to findings in 261 cases of 47 families published in the literature. It was concluded that HNPP is a nosological entity characterized by the following diagnostic criteria: (1) an autosomal dominant inheritance; (2) the clinical presentation of a recurrent mononeuropathy simplex or multiplex, frequently related to an inadequate trauma to peripheral nerves; (3) a significant slowing of motor and sensory conduction velocity in clinically affected, but also in clinically unaffected nerves; (4) characteristic morphological findings in sural nerve biopsy featuring tomaculous swellings of myelin sheaths, transnodal myelination and segmental demyelination. The pathogenesis of HNPP is not clear. Hypothetical explanations of the pathogenesis of HNPP are discussed.In memory of Albert Bischoff (1921–1981), Professor of Neurology, University of Berne     

结核分枝杆菌热休克蛋白65与汉坦病毒G2糖蛋白融合基因真核表达载体的构建与表达

目的 构建以结核分枝杆菌热休克蛋白65（HSP65）和汉坦病毒G2糖蛋白重组融合基因为基础的真核表达载体。为进一步研究其在结核病和出血热防治中的作用奠定基础。方法 采用聚合酶链反应从结核分枝杆菌H37Rv株基因中．扩增出HSP65的编码基因，从T-H8205G2质粒扩增出G2糖蛋白的编码基因．经相应限制性核酸内切酶消化后．插入真核表达载体pcDNA3．1／His，并将此重组质粒通过脂质体介导转染NIH3T3细胞．用SDS-PAGE、免疫组织化学及免疫荧光方法分别测定表达产物的相对分子量及特异性。结果 获得正向插入的pcDNA3.1/ His-HSP65-G2重组表达质粒，表达产物的相对分子量为105kD．与理论预期大小一致．并且可与汉坦病毒H8205株的腹水抗体和HSP65单抗起特异反应。表明构建的pcDNA3．1／His-HSP65-G2能在哺乳动物细胞中表达并具有抗原性。结论 编码HSP65和G2抗原基因的重组基因的真核表达载体构建成功。     

松口蘑发酵菌丝体糖蛋白MTS03对人乳腺癌细胞MCF-7的体外增殖抑制作用研究

 目的研究松口蘑发酵菌丝体糖蛋白MTS03的理化性质及抗肿瘤作用。方法采用SDS-PAGE鉴定MTS03的纯度及相对分子质量,氨基酸自动分析仪分析其中的氨基酸组成;MTT法研究糖蛋白MTS03体外对人乳腺癌细胞MCF-7的增殖抑制率,同时考察MTS03对人正常肝细胞L-02的杀伤率,评价其安全性。结果SDS-PAGE电泳鉴定为单一组分,测定其相对分子质量约为2.37×10⁴,经氨基酸自动分析仪分析含有17种氨基酸;MTS03对MCF-7具有直接的细胞毒作用,在一定的浓度范围内呈现量效和时效关系,MTS03对MCF-7作用24,48,72h的IC₅₀分别为25.81,12.46,8.86mg·L^-1,而对正常肝细胞的IC₅₀分别为1366.49,190.60,95.58mg·L^-1,对MCF-7的治疗指数TI分别为54.16,18.74,12.25。结论MTS03对于体外培养的乳腺癌细胞具有良好的直接杀伤作用,而对正常肝细胞的毒性较小,可能是一种高效低毒的生物活性物质。