Ubiquitin-positive achromatic neurons in corticobasal degeneration

A 66-year-old woman presented with an alien limb syndrome without dementia. The course of her illness was unremitting and at autopsy 6 years later her diagnosis was confirmed as corticobasal degeneration without Alzheimer-type pathology. Although the presence of ballooned achromatic cortical neurons and cell loss from the substantia nigra distinguishes such patients, the site and density of achromatic neurons has not previously been quantified. We show that immunohistochemistry for the cell stress protein ubiquitin selectively stains these achromatic neurons, whereas they do not stain for abnormally phosphorylated tau protein. Phosphorylated neurofilament antibodies recognise both ballooned and non-ballooned neurons. In this case, high densities of ubiquitin-positive ballooned neurons were found in frontal cortical regions with the highest densities in layers V and VI of the anterior cingulate cortex. In addition, high densities of ubiquitin-positive ballooned neurons were found in the insular cortex, claustrum and amygdala. These results confirm past reports of frontal pathology, but show that there is also considerable pathology in insular and parahippocampal cortical regions and some subcortical regions. Our findings suggest that the distribution and staining characteristics of ballooned neurons in corticobasal degeneration may help to differentiate these cases pathologically, while the absence of dementia appears to be an important clinical criterion.     

Quantitative assessment of technetium-99m methoxyisobutylisonitrile planar perfusion heart studies: application. of multivariate analysis to patient classification

A quantified evaluation of planar cardiac perfusion scintigrams (the objective of the study), obtained using technetium-99m methoxyisobutylisonitrile (MIBI) was performed on the basis of an analysis of circumferential profile curves, representing the perfusion as seen in three typical projections. The analysis involved the curves obtained both at rest and after stress, and was based on a comparison of their shape (trend) with the normal trend (normative evaluation). The latter was obtained by means of an original method of iterative fitting of individual curves into the database. The base consisted of curves recorded in 53 patients (separately in males and females) with normal perfusion of the left ventricle (group I, the reference group). A group of 90 patients suspected of having coronary artery disease (group II) was subdivided into two subgroups on the basis of coronary arteriography: (a) those with and (b) those without critical stenosis of at least one artery. Profile curves characterising the LV perfusion were obtained at rest and after stress. Defects of perfusion were quantified by comparison of individual curves with the normal trends. By means of multivariate analysis it was demonstrated that vectors of mean values characterising the scintigraphically assessed defects of LV perfusion in the two subgroups of group II differed very significantly (P<10^–5). Applying methods of discriminant analysis, a classification of patients from group II was performed into those with probable defects of perfusion and those free of such defects. The sensitivity, specificity and accuracy of diagnosis of coronary ischaemia, based on quantified planar^99mTc-MIBI scintigraphy, reached 86%, 87% and 87%, respectively.     

Functional imaging of the brain using single photon emission computerized tomography (SPECT)

Summary The use of tracers is an important technique available for studying cerebral function. Changes in signal are large, but as a result of its photon limited nature, the measurement of this signal is limited: spatially, temporally and in terms of accuracy. The most commonly used single photon (SPECT) system (as apposed to positron) is that with a rotating gamma camera, although multi-headed devices and special purpose rings are now also commonly available. The problems of obtaining good functional information are however identical. Firstly the devices need to be optimised in terms of resolution and sensitivity. Secondly several sources of error, notably those associated with scatter, attenuation and limited spatial resolution, need to be corrected, with the aim of obtaining quantitative estimates of radioactivity concentration. Finally such quantitative estimates need to be converted into meaningful estimates of physiological variables by use of an appropriate model. The general aim of many SPECT measurements is to estimate blood flow for example using Tc-99m labelled HMPAO as a tracer. Good results have been obtained in many clinical conditions: stroke, dementia, tumour and epilepsy, for example. Many other tracers are also available, for example to measure density of receptor sites. The use of SPECT in conjunction with other techniques after image registration is suggested as being an essential tool in extracting maximal clinical information.     

Number and size spectra of non-myelinated axons of human premolars

The aim of this study was to determine the number and size of apical non-myelinated (C) axons of healthy human premolars. The material was derived from a large collection of specimens prepared for a previous quantitative investigation on the myelinated (A) axons of human premolars. A total of 16 teeth (six maxillary first and five each of mandibular first and second premolars), removed from adolescents for orthodontic reasons, were used. Root discs of about 0.6 mm thickness were prepared at about 2 mm cervical to the root apex and processed for light and electron microscopy. The number of non-myelinated axons was determined by taking a total census of such fibres that could be identified and reconstructed by standardized composite electron micrographs from each root disc. The measurement of axons was done on a statistically representative sample of axons (n=1810) using an electronic image processing unit. The 16 teeth had an average of 2000 ± 1023 non-myelinated axons at the juxta-apical level (range 534–3912). The average diameter of the non-myelinated axons was found to be 0.5 ± 0.4 m (range 0.05–2.4 m).     

Number and size-spectra of myelinated nerve fibers of human premolars

Summary The primary objective of this study was to determine the number and size of myelinated nerve fibers at the subcervical, midroot and juxta-apical levels of human premolars. Sixty-seven healthy premolars extracted from adolescents were utilized. Root-discs were prepared from the three sites and processed for light and electron microscopy. The myelinated nerve fibers were counted from semithin sections using a sampling microscope. The measurements were taken from composite electron micrographs using an electronic image processing unit. A total of 1883 myelinated axons from seven mandibular second premolars was gauged. The 67 teeth had an average of 312±149 myelinated nerve fibers at the juxta-apical level (range 18 to 728). The contra- and ipsilateral differences in means among the four groups of premolars were not significant (P>0.05). The number of nerves increased significantly (P<0.05) toward midroot and subcervical (P<0.001) levels in all groups. The average neural diameter was 3.51.0 m at the juxta-apical level, and the between-teeth difference in mean was found to be significant (P<0.01). There was no decline (P>0.05) in the diameter of myelinated nerve fibers toward midroot and subcervical levels.     

Quantitation of Antibodies to Toxoplasma gondii in Swine Sera by Enzyme-Linked Immunosorbent Assay

Abstract

An enzyme-linked immunosorbent assay (ELISA) was developed for quantitation of antibodies to Toxoplasma gondii in swine sera. Because a commercial anti-swine IgG conjugate was directed also against swine IgM, the conjugate was absorbed with the IgM fraction to eliminate the interference of naturally occurring IgM antibodies that appeared consistently in sera collected from slaughtered pigs at an abattoir. The ELISA values of 0.2 or more observed in most of the sera successfully decreased to less than 0.2 by the use of absorbed conjugate. An attempt to use a protein A conjugate has failed. Evaluation of this system by comparing it with the latex agglutination test provided a high significant correlation, indicating its usefulness for serodiagnosis of swine toxoplasmosis.     

基于~1H-MRS与LCModel软件检测健康青年人服用磷酸可待因后前额叶及海马代谢物变化

目的通过绝对定量分析健康青年人服用磷酸可待因后双侧前额叶及海马代谢物浓度变化,为下一步研究青年人可待因慢性成瘾机制提供前期实验数据参考。材料与方法正常健康青年志愿者20名,男10名,女10名,年龄18~30岁,平均(24.9±1.9)岁,均为右利手,无神经、精神疾病病史。采用GE 1.5 T Signa HDX超导MR扫描仪,单体素1H-MRS PRESS序列,TR 3000 ms,TE 30 ms,矩阵256×128,NEX为1,感兴趣区置于双侧前额叶及双侧海马,大小2 cm×2 cm×2 cm,采集服药前及口服60 mg磷酸可待因后1.0~2 h内的实验数据,利用LCModel软件对采集的数据进行后处理及定量,用SPSS19.0配对样本t检验进行统计学分析。结果服用磷酸可待因后左前额叶脑代谢物发生改变,甘油磷酸胆碱(GPC)浓度增加0.254 mmol/L、肌醇(Ins)浓度下降0.988 mmol/L,差异有统计学意义(n=13,P0.05;n=12,P0.05);右前额叶、双侧海马的代谢物浓度改变无统计学差异。结论口服磷酸可待因能引起左前额叶代谢物的改变,GPC浓度上升与Ins浓度下降可能与急性用药后前膜递质释放减少有关。     

应用磁共振软骨T2图成像定量测定退变兔膝关节软骨

目的:探讨T_2图成像技术在检测关节软骨退变早期的应用价值。材料和方法:选用20只新西兰大白兔,随机分为甲、乙、丙、丁四组。甲组左膝关节行常规磁共振成像,扫描结束后即刻处死,取股骨髁软骨行苏木素和伊红染色(HE)、阿利辛兰染色(AB)及蛋白多糖含量测定。乙、丙、丁各组每只兔左膝关节内注射0.2ml(10U)木瓜蛋白酶。乙、丙、丁各组于注射木瓜蛋白酶前及注射后分别于24h、48h、72h先行相同常规磁共振成像及T_2图成像,取感兴趣点,测定关节软骨T_2弛豫时间值并取平均值,对每组数据进行t检验。扫描结束后处死动物,取左膝股骨髁部软骨行大体观察、HE、AB染色及蛋白多糖含量测定。结果:注射木瓜蛋白酶后24h、48h,蛋白多糖含量与甲组比较,统计学均有差异(P=0.048和0.045,P<0.05),注射后72h,统计学没有差异(P=0.455,P>0.05)。T_2图像软骨色阶由注射前的黄红色转为注射后的红色,注射木瓜蛋白酶后24h、48h、72h的T_2弛豫率分别升高了30.14ms,20.92ms和16.17ms。经t检验后,注射后24h与注射前差异具有统计学意义(P= 0.04,P<0.05)。结论:本研究采用的T_2图成像技术能够通过定量检测T_2弛豫时间值反映软骨退变早期软骨内的生化改变。     

Quantitative analysis of mannitol polymorphs. FT-Raman spectroscopy

Mannitol is a polymorphic excipient which is usually used in pharmaceutical products as the beta form, although other polymorphs (alpha and delta) are common contaminants. Binary mixtures containing beta and delta mannitol were prepared to quantify the concentration of the beta form using FT-Raman spectroscopy. Spectral regions characteristic of each form were selected and peak intensity ratios of beta peaks to delta peaks were calculated. Using these ratios, a correlation curve was established which was then validated by analysing further samples of known composition. The results indicate that levels down to 2% beta could be quantified using this novel, non-destructive approach. Potential errors associated with quantitative studies using FT-Raman spectroscopy were also researched. The principal source of variability arose from inhomogeneities on mixing of the samples; a significant reduction of these errors was observed by reducing and controlling the particle size range. The results show that FT-Raman spectroscopy can be used to rapidly and accurately quantitate polymorphic mixtures.