Contraluminal transport of organic cations in the proximal tubule of the rat kidney

In order to study the quantitative structure/activity relationship of organic cation transport across the contraluminal side of the proximal renal tubule cell, the stopped-flow capillary microperfusion method was applied and the inhibitory potency (apparent K _i values) of different homologous series of substrates against N ¹-[³H]methylnicotinamide (NMeN⁺) transport was evaluated. Aniline and its ring- or N-substituted analogues as well as the aminonaphthalines do not interact with the contraluminal NMeN⁺ transporter except for the quaternary trimethylphenylammonium and pararosaniline, which bear a permanent positive charge, and for 1,8-bis-(dimethylamino)naphthaline, which forms an intramolecular hydrogen bond. If, however, one or more than one methylene group is interposed between the benzene ring and the amino group, the compounds interact with the contraluminal NMeN⁺ transporter in proportion to their hydrophobicity parameter, i.e. the octanol/water partition coefficient (log octanol). The catecholamines and other hydroxyl-substituted phenylethyl analogues also follow this rule. In addition, the N-heterocyclic pyridine, quinoline, isoquinoline and acridine analogues also interact with the contraluminal NMeN⁺ transporter, when their pK _a values are higher than 5.0, and, an inverse correlation between pK _a and log K _{i, NMeN} was observed. An exception to this rule are those hydroxy compounds of pyridine, quinoline and isoquinoline that show tautomerism. These compounds slightly inhibit NMeN⁺ transport despite low pK _a values. The quaternary nitrogen compounds of aniline and the N-heterocyclic analogues, as far as tested, all interact with the contraluminal NMeN⁺ transporter in relation to their hydrophobicity. The data indicate that the contraluminal NMeN⁺ transporter interacts with N-compounds according to their hydrophobicity and/or according to their basicity (affinity to protons). The reason for deviation of the aniline analogues and the OH-tautomeric heterocyclic N-compounds from this behaviour is discussed.     

Secretin and VIP-stimulated adenylate cyclase from rat heart

The exact positions of microelectrodes used to measure thePO₂ in the cerebral cortex of the rat were determined by staining the tissue with Alcian Blue. The measurement sites were subsequently located under a light microscope and correlated with the capillary and cellular arrangement of the cortex. The microelectrodes used for thePO₂ measurements were made of gold glass fibers; the Alcian Blue was injected hydrostatically through a micropipette attached to thePO₂ microelectrode. The sites where dye had been deposited were seen under a light microscope as green blue spots about 100 m in diameter. The capillaries were visualized by silver nitrate perfusion. Differences between the localPO₂ values in the neo- and the archeocortex were found. In the neocortex the meanPO₂ was 31 mm Hg, capillary volume 1.6%, capillary surface area 980/mm², capillary length 13.5/mm; whereas in the archeocortex these values where 21 mm Hg, 0.9%, 820/mm² and 9.4/mm respectively. These data indicate a relationship between the microcirculatory transport system and the local oxygen tension and provide further evidence that the meanPO₂ level tends to decrease when moving from the surface into the archeocortex.Supported by the Deutsche ForschungsgemeinschaftReported in part at the 3rd Symposium of ISOTT, Cambridge, GB, 1977; and at the 27th International Congress of Physiological Sciences, Paris, France, 1977     

A revised cytoarchitectonic map of the neocortex of the rabbit (Oryctolagus cuniculus)

Summary The cytoarchitectonic parcellation of the rabbit's neocortex has been investigated in 6 hemispheres which had been fixed by perfusion, embedded in paraffin and sectioned at either 9 m or 20 m in various planes. In addition to the classical method of microscopic observation, and automatic scanning procedure using an image analyser for measuring grey level indices was employed. By printing computer plots of various ranges of grey level indices, this method permits visualization of structural differences between various cytoarchitectonic fields. By evaluating the plots, cytoarchitectonic maps can be constructed which are based on objective data and therefore less influenced by subjective judgment than the maps obtained with the classical method. — In some regions the results based on the quantitative method are in agreement with the commonly used maps of Rose (1931), and in other regions widely at variance. It is shown, for instance, that the area striata as defined by Rose (1931) is composed of two distinct fields, viz. areas Oc 1 and 2, which are separated from each other in the rostro-caudal direction. These and other findings are described in detail, compared with the observations of Rose (1931), and related to the literature on functional localization in the rabbit's neocortex. Attention is drawn to the fact that the results obtained in 6 hemispheres leave no doubt that individual variations in size and shape of the entire hemisphere as well as of the various cytoarchitectonic fields do occur, and will have to be taken into account if cytoarchitectonic maps such as those published in the present paper are to be used in the context of experimental work.     

Quantitative distribution of rat brain monoamine oxidase A by [14C]clorgyline autoradiography

The distribution of functionally active monoamine oxidase type A (MAO-A) was investigated by in vivo quantitative autoradiography using [¹⁴C]clorgyline in normal, conscious rat brain. [¹⁴C]clorgyline was synthesized by the methylation reaction of N-desmethylclorgyline using [¹⁴C]methyliodide. Sixty minutes after [¹⁴C]clorgyline administration (1.58 MBq/animal i.v.), the brains were removed and prepared for autoradiography by washing the brain sections with 5% trichloroacetic acid solution to remove the nonbinding free tracer. The amount of MAO-A was calculated from the regional acid-insoluble tissue radioactivity and the specific activity of the tracer. The highest amount of MAO-A (5.84 nmol/g tissue) was found in the locus coeruleus. The interpeduncular nucleus, habenular nucleus, fasciculus retroflexus, and solitary tract nucleus possessed over 1.6 nmol/g tissue of MAO-A. Among 23 regions of interest, the lowest amount of MAO-A (0.37 nmol/g tissue) was found in the globus pallidus. The findings of this study suggest that the pattern of MAO-A parallels both in neuroanatomical distribution and in density that of norepinephrine and serotonin innervation. The MAO-A concentration was, however, relatively low in the dopamine-related areas. This corresponded to the previous results obtained by histochemical analysis. In addition, among the white matter structures, a high amount of MAO-A was found specifically in the fasciculus retroflexus.     

Anterior tibial artery and its actual projection on the lateral aspect of the tibia: a cadaveric study

The anterior tibial artery (ATA) is at risk of injury during high tibial osteotomy, Ilizarov wire placement, pin placement in external fixation, or proximal locking screw insertion, as the artery is not visualized intraoperatively. The ATA is anchored to the oval foramen of the interosseous membrane on the proximal tibia by the deep fascia and recurrent genicular vascular branches. Segment 1 (from the bifurcation of the popliteal artery to the level of the interosseous foramen) and the proximal part of segment 2 (from the interosseous foramen to the level where the artery crosses the anterior border of the tibia) may be damaged when pin, wire or screw placement is directed posterolaterally at that level. Distally, a straight mediolateral pin or Ilizarov wires may lacerate the artery. Segment 2 of the ATA descends against the interosseous membrane in its proximal part, which is projected on the posterior third of the tibia relative to the sagittal plane; in its middle part, it runs close to the lateral cortex of the tibia, it is projected on the middle third of the tibia; in its distal part it runs gradually towards the anterior third of the tibia and contacts with the anterior third of the tibial cortical surface. This information may help reduce risk of injury to the ATA during high tibial osteotomy, external fixation and pin placement or insertion of locking screws.     

核素唾液腺显像的定量分析软件开发与应用

目的开发定量分析唾液腺功能及其图象显示的程序。方法应用Aladdin语言编写ROI模块提取各唾液腺及甲状腺区域的摄取数据及本底,用曲线函数生成各ROI的时间-放射性活度曲线,对曲线数据处理,计算唾液腺摄取、排泌指数等指标,然后把图象、曲线及计算结果显示在一个模板内。结果同一幅输出界面图可显示唾液腺的形态、ROI示意图、唾液腺的摄取与排泌功能数值及其曲线,以标准化的显示格式提供给临床医师。不同操作者处理同一资料的功能指标结果基本一致,软件计算的功能指数与图象显示结果及临床诊断基本相符。结论开发的唾液腺功能定量分析软件,程序运行稳定,结果重复性好。     

Real-time PCR method for the quantitative analysis of human T-cell receptor gamma and beta gene rearrangements

Analyzing the status of T-cell receptor (TCR) gene rearrangements has been an essential part of deciphering the stages of thymocyte development, understanding the β vs. γδ lineage decision, and characterizing T-cell leukemias. Methods such as PCR and quantitative Southern blotting provide useful information, but also have significant shortcomings such as lack of quantitation in the case of PCR and technical challenges in the case of Southern blotting. Here we describe a real-time PCR method that overcomes many of these shortcomings. This new method shows comparable results for the fraction of unrearranged TCRγ and TCRβ genes in human thymocytes and peripheral blood T cells as Southern blotting, and has the advantages of being simple to perform, highly quantitative, and requiring nanogram quantities of DNA. We also describe a real-time PCR method to quantitate T-cell receptor excision circles formed during TCRβ rearrangements.     

化合物构效关系的模糊极小极大神经网络识别研究

应用模糊极小极大神经网络研究了化合物复杂结构和性能（QSAR）之间的关系，用该法进行几组化合物致癌的识别，结果优于线性回归的方法，对此作出一些分析。     

微量酶联夹心杂交法定量检测hIL-8 mRNA PCR扩增产物

目的:建立一种灵敏度高、特异性强、定量、精确、操作简便的微孔板酶联夹心杂交技术,定量检测人IL-8 mR-NA。方法:针对人IL-8 mRNA逆转录聚合酶链反应产物一条链的不同区域序列设计一对特异探针,其中一条为捕获探针,5′端用活性氨基修饰,与微量DNA结合板表面的NOS基团共价结合,“竖直”地包被在微孔板内;另一条检测探针的3′端标记生物素,和辣根过氧化物酶结合。提取人外周血单个核细胞总RNA,进行RT-PCR扩增IL-8 mRNA,双链DNA产物经热变性后加入已包被捕获探针的微孔板内进行杂交,加入检测探针与已杂交的产物结合,经亲和素-辣根过氧化物酶系统检测杂交信号。结果:该法灵敏度为检出16个循环的PCR产物、5×103个PBMCs中的IL-8 mRNA、检测PCR终产物的最高稀释倍数为1∶256阳性;特异性试验非目的扩增片段酶联杂交检测未检出阳性杂交信号;精密度试验CV为5.2%。结论:该方法具有操作简便、灵敏度高、特异性强等优点,适合IL-8 mRNA PCR扩增产物的定量检测。