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The actin-binding protein ezrin is associated with cellular shape changes, motility, tumor invasion, and lymphocyte activation. We have earlier shown that ezrin immunoreactivity (IR) is faintly present in normal astrocytes but increased in malignant human astrogliomas. We studied the role of ezrin in astrocyte activation, applying immunostaining on serial paraffin sections from human autopsied brain tissues (51 cases). Cerebral HIV infection was chosen as a model displaying consistent exemplary astrocyte activation. Semiquantitative ezrin-IR was compared with the common glial markers GFAP, ferritin, and HLA-DR in relation to clinical and morphologic criteria of HIV encephalopathy. In all cases with HIV infection, GFAP-, HLA-DR-, and ferritin-IR were elevated in comparison to normal brain tissues. In contrast, high ezrin-IR in HIV infection strictly correlated with additional HIV encephalopathy. HIV encephalopathy with particularly high ezrin-IR was correlated with neuronal apoptosis (TUNEL). Combined ezrin-IR and GFAP-IR thus reveals 2 distinct states of astrocytic activation. Normal ezrin-IR, when paralleled by upregulated GFAP, reflects astroglial activation not associated with neuronal apoptosis. High ezrin-IR indicates specific astrocyte stressors related to cellular damage within the central nervous system. Ezrin-IR might also provide a diagnostic tool for the classification of HIV encephalopathy.

Purpose: To investigate whether histomorphology of tumor cell nuclei has a significant and independent relation to survival time of patients with glioblastomas. Experimental Design: Seventy-two tumors from 72 patients were investigated by means of digital image analysis. Proliferating and nonproliferating nuclei were separately measured and parameters of nuclear size, shape, texture, and spatial relationships (topometric parameters) were detected. Survival analysis was done regarding morphometric data together with the patients' age, the amount of resection (total or subtotal), and the classification of the tumor as a “primary” (de novo) or “secondary” glioblastoma. Results: The overall relation of all morphometric data to the time of survival was highly significant (Cox analysis, P < 0.0001). Apart from the extent of surgical resection, parameters of nuclear shape and topometric variables, such as the distance between two nuclei lying nearest to each other, showed an independent and significant relation to survival time. The patients' age had also a significant but comparably slight relation to survival time. Conclusions: The morphology of tumor cell nuclei, as represented by morphometric data, shows a significant relation to survival time of patients with glioblastomas. This relation is statistically independent from the amount of surgical resection, from the patients' age and from the classification of the glioblastoma as being primary or secondary. The results support the view that histomorphometry of tumor cell nuclei is a valuable prognostic marker for patients with glioblastomas. We believe that such a marker ought to be incorporated into the formation of individual therapeutic decisions.

In contrast to the growing interest in proton-MR-spectroscopy (1HMRS) for preoperative examination of patients with brain tumors, there is nearly no knowledge about a correlation between data from 1HMRS and histomorphology as confirmed by quantitative morphological methods. Whether a correlation can be confirmed between data from 1HMRS and quantitative histomorphology of glioblastomas representing the most frequent type of brain tumors was investigated in the present study. Furthermore, it was of interest, whether correlations between spectroscopic data and histomorphology can be confirmed for proliferating and non-proliferating tumor cell nuclei independently. Using stringent inclusion criteria for this study, 24 patients were investigated by means of preoperative 1HMRS and by means of digital image analysis of paraffin sections from the surgical specimen. Proliferating and non-proliferating tumor cell nuclei were investigated separately in the region with the highest proliferative activity in each tumor using immunohistological staining for the proliferation marker Ki67. Main results showed highly significant correlations between total creatine and variables of nuclear size, as well as correlations between choline and variables of nuclear shape. These results were confirmed for both proliferating and non-proliferating tumor cell nuclei. A significant correlation between N-acetyl-aspartate level and topometric variables (number of neighbors per nucleus, variables describing distances between tumor cell nuclei) was confirmed for proliferating tumor cell nuclei. Discriminant analysis provided a good separation of cases with high and with low values for these spectroscopic variables based on histomorphometric data. In conclusion, the results confirm a direct correlation between data from preoperative 1HMRS and histomorphological characteristics of glioblastomas supporting the biological relevance of spectroscopic data for the examination of brain tumor patients.

A comparison between data from proton-MR spectroscopy (1HMRS) and quantitative histomorphology of tumor cell nuclei in gliomas has not been reported up to now. Therefore, the question must be answered, if there are any significant correlations between histomorphology of gliomas and quantitative data from 1HMRS concerning tissue metabolites. Surgical glioma specimen (glioblastomas, astrocytomas, oligodendrogliomas) from 46 patients with tumor grades II–IV according to WHO have been evaluated by means of a digital image analysis system using Ki-67-immunostained paraffin sections. Nuclear density, Ki-67-proliferation index, nuclear area and shape variables (roundness factor, Fourier-amplitudes) have been determined from 200 randomly selected tumor cell nuclei in each tumor specimen. These data have been correlated with preoperative data from 1HMRS. A positive correlation between Fourier-amplitudes, choline peak and lipide peak was observed, as well as a negative correlation between these variables and the nuclear roundness factor. This result indicates higher choline and lipide peaks with increasing irregularity of nuclear outlines. Proliferation index Ki-67 was positively correlated with the lipide peak, nuclear density showed a positive correlation with the choline peak. Glioblastomas (n = 29) showed an additional positive correlation between mean nuclear size and total creatine. Anaplastic gliomas (n = 12) showed a positive correlation between lactate peak and the standard deviation of the nuclear roundness factor. Further multivariate analyses have shown, that for the present collective of 46 cases, histometric variables have a higher significance than spectroscopic data for the differentiation of the different tumor grades. These results verify a significant correlation between preoperative data from 1HMRS and histomorphology of tumor cell nuclei in gliomas, supporting the biological significance of both histomorphometry and 1HMRS for the evaluation of these tumors.

Object. In this study, 1H magnetic resonance (MR) spectroscopy was prospectively tested as a reliable method for presurgical grading of neuroepithelial brain tumors. Methods. Using a database of tumor spectra obtained in patients with histologically confirmed diagnoses, 94 consecutive untreated patients were studied using single-voxel 1H spectroscopy (point-resolved spectroscopy; TE 135 msec, TR 1500 msec). A total of 90 tumor spectra obtained in patients with diagnostic 1H MR spectroscopy examinations were analyzed using commercially available software (MRUI/VARPRO) and classified using linear discriminant analysis as World Health Organization (WHO) Grade I/II, WHO Grade III, or WHO Grade IV lesions. In all cases, the classification results were matched with histopathological diagnoses that were made according to the WHO classification criteria after serial stereotactic biopsy procedures or open surgery. Histopathological studies revealed 30 Grade I/II tumors, 29 Grade III tumors, and 31 Grade IV tumors. The reliability of the histological diagnoses was validated considering a minimum postsurgical follow-up period of 12 months (range 12–37 months). Classifications based on spectroscopic data yielded 31 tumors in Grade I/II, 32 in Grade III, and 27 in Grade IV. Incorrect classifications included two Grade II tumors, one of which was identified as Grade III and one as Grade IV; two Grade III tumors identified as Grade II; two Grade III lesions identified as Grade IV; and six Grade IV tumors identified as Grade III. Furthermore, one glioblastoma (WHO Grade IV) was classified as WHO Grade I/II. This represents an overall success rate of 86%, and a 95% success rate in differentiating low-grade from high-grade tumors. Conclusions. The authors conclude that in vivo 1H MR spectroscopy is a reliable technique for grading neuroepithelial brain tumors.

Proton MR spectroscopy ((1)H MRS)-visible total choline-containing compounds (tCho-compounds) are derivatives of membrane phospholipids and, in part, may act as a long-term second-messenger system for cellular proliferation. Experimental evidence suggests increasing concentrations of tCho-compounds during cellular proliferation. The present study was conducted in order to test the hypothesis that in vivo measurements of tCho-concentrations using (1)H MRS allow assessment of the proliferative activity of neuroepithelial brain tumors presurgically. Single-voxel (1)H MRS (PRESS, TR 1500 ms, TE 135 ms) was performed in 101 patients with neuroepithelial brain tumors prior to surgery and 19 healthy volunteers. Histological diagnoses were confirmed postsurgically according to the WHO classification. Measured tCho-compound signal intensities were corrected for coil loading, numbers of acquisitions and voxel size, and tCho concentrations calculated as institutional arbitrary units. They were matched with the mean immunohistochemical marker of cell proliferation, the Ki-67 (MIB.1) labeling index, using correlation analysis according to Spearman. Compared with low-grade tumors (i.e. WHO grade I/II) and normal white brain matter, high-grade tumors (i.e. WHO grade III/IV) revealed significantly (p < 0.05) elevated labeling indices paralleled by increasingly elevated tCho-concentrations. In contrast tCho-concentrations in low-grade tumor did not differ significantly from physiological values. A highly significant positive correlation (p < 0.0001, r(2) = 0.81) was found between the tCho-concentration and the labeling index. It was concluded that the determination of tCho-concentrations using in vivo (1)H MRS could provide a novel and noninvasive assessment of the proliferative activity of neuroepithelial brain tumors, pointing at (1)H MRS as a useful method for differentiating proliferating from non-proliferating tissues. Hence, potential indications for the clinical application of (1)H MRS are grading tumors presurgically, early detection of anaplastic transformation, and monitoring treatment.

Lipofuscin has been of interest in dementia of Alzheimer’s type (DAT), because of the hypothesis that dementia is a kind of premature ageing. The results of studies on intraneuronal lipofuscin content in DAT are inconsistent. At present no data on intraneuronal lipofuscin in dementia with Lewy bodies (DLB) are available. The amount of lipofuscin in the neurons of the inferior olivary nucleus of 10 brains of DLB patients, 10 of age-matched DAT patients, and 10 of age-matched controls has been measured microfluorometrically. The intraneuronal lipofuscin content in the DLB patients was significantly lower than in the DAT patients and controls. This finding may indicate a general alteration of neuronal cell metabolism in DLB, e.g. a defect of mitochondrial DNA, not confined to neurons developing Lewy bodies.

Hintergrund: Die juvenile neuronale Zeroidlipofuszinose (JNCL) ist fur den Augenarzt wichtig, da die Erstsymptome haufig das Auge betreffen und eine fruhe Diagnosestellung ermoglichen. Die Erkrankung beginnt im 4. bis 10. Lebensjahr meist mit einem Visusabfall infolge einer Makulopathie, gefolgt von einer rasch fortschreitenden Degeneration von Netzhaut und Pigmentepithel und fuhrt binnen ca. 1 – 3 Jahren zur Erblindung. Danach stehen neurologische Veranderungen (motorische und intellektuelle Defizite) im Vordergrund; der Tod tritt meist bis zur 3. Lebensdekade ein. Methode: Anhand eines Falls von 2 Schwestern werden Befunde und diagnostisches Vorgehen unter Berucksichtigung elektrophysiologischer und erganzender Untersuchungen (Blutausstrich, Histologie) erlautert. Ergebnisse: Bei der alteren (jungeren) Schwester traten mit 8(6) Jahren erste Sehprobleme auf; bei der Untersuchung betrug die Sehscharfe Lichtprojektion (0,05). Morphologisch zeigte sich eine tapetoretinale Degeneration mit Papillenatrophie, Gefasverengung, Pigmentepitheliopathie und Bulls-eye-Makulopathie. Im ERG waren bei der alteren Schwester lediglich Restpotentiale ableitbar, bei der jungeren war es deutlich vermindert (skotopisch mehr als photopisch). Im Blutausstrich fanden sich vakuolisierte Lymphozyten, bei der histologischen Untersuchung der Bindehautbiopsie zeigten sich Einschlusse vom Typ der kurvilinearen Korper sowie Fingerprint-Strukturen. Schlusfolgerungen: Trotz einer starken Sehbeeintrachtigung mit bereits ca. 6 – 7 Jahren wird die Diagnose JNCL haufig erst Jahre spater, wenn neurologische Symptome wie Grand-mal-Anfalle hinzukommen, gestellt. Bei plotzlichem Visusabfall im entsprechenden Alter und elektrophysiologisch gesicherter tapetoretinaler Degeneration sollten daher zumindest ein peripherer Blutausstrich und ggf. eine Biopsie veranlast werden.

The brainstems of ten patients with Alzheimer's disease were examined with specific silver impregnations for beta-amyloid deposits, neurofibrillary tangles, neuropil threads and neuritic plaques. The results show a selective and focal involvement of brainstem nuclei, which are diffusely connected to the cortex or are neuronally connected with other damaged subcortical and cortical regions. Therefore, it is concluded that neuronal connectivity plays an important role in the pathogenesis of Alzheimer's disease lesions. This may be due to the intraneuronal transport of beta-amyloid precursor protein. There was a local association between neurofibrillary tangles and neuropil threads, but not between beta-amyloid structures on the one hand and neurofibrillary structures on the other hand. Neuritic plaques were rarely found.

The present study reports a new method for the densitometric measurement of the intensity of immunohistochemical reactions. This method is based on a programm for the Kontron VIDAS image analysis system and has been designed for the measurement of small differences in the relative intensity of immunohistochemical reactions. Immunohistochemistry was performed with the avidin-biotin-peroxidase complex and diaminobenzidine-HCl and H2O2 for enzyme visualization. Several methods for shade correction and image processing were elaborated. The study was carried out on gerbil Purkinje cells using monoclonal antibodies raised against calbindin D28k. Prerequesites of correct measurement were standardized preparation, i.e., identical thickness of the paraffin sections, identical performance of immunohistochemistry, and avoidance of any counterstaining. The evaluation of small intensity differences of immunohistochemical reactions was found to be feasible either by substractive shade correction and standardized normalization or by shade correction by division by a reference image and standardized thresholding. Small differences in antigen concentration were not detectable without additional image processing.

The lipofuscin of pyramidal cells in each hippocampal subfield of each of seven human autopsy cases without brain disease at the age of 3-12 months (infants) and of 17-23 years (young adults) was comparatively investigated at the electron microscopic level. In infant pyramidal neurons of the hippocampal subfields CA 1, CA 2, CA 3 and CA 4 O-2, very small lipofuscin particles were observed. The lipofuscin composition showed a slightly larger granular component compared to the vacuolar component with one or two small lipid droplets. No obvious ultrastructural variability of lipofuscin granules was observed. The CA 1 lipofuscin in young adults consists of larger particles than in infants, but no obvious difference in the composition of granular and vacuolar components from the infant lipofuscin was seen. The amount of lipofuscin in CA 1 strongly increased in young adults compared to infants and appeared in a perinuclear distribution. In young adults, in contrast to the infant group, the amount of lipofuscin in the subfields CA 2, CA 3 and CA 4 was significantly higher than in CA 1. In CA 2, CA 3 and CA 4 pyramidal neurons, the vacuolar component was significantly larger than the granular component. The similarity of infant hippocampal lipofuscin patterns in all subfields is discussed as a state of immaturity. To explain the observed differences between the CA 1 and the other subfields during neuronal development, as shown in the young adult group, several factors are discussed: the effects of cell specific metabolism, cellular functional activity, cytoprotective mechanisms and effects of efferent and afferent pathways connected with the subfields.

Background and Purpose The sensitivity and validity of serum neuron-specific enolase as a marker of brain injury were tested after global cerebral ischemia. Methods Sixty-nine Mongolian gerbils were perfusion fixed after variable reperfusion after 5-minute (group 1) or 15-minute (group 2) bilateral carotid occlusion. Neuron-specific enolase was analyzed by an enzyme immunoassay in serum of control, sham-operated, and ischemic animals before euthanasia and in nonischemic gerbil brains. Brains were processed for histology, immunohistochemistry, and morphometric evaluation of ischemic neuronal damage. Results After cerebral ischemia, loss of neuronal immunoreactivity was closely associated with increased neuron-specific enolase serum levels, which were significantly elevated by 24 hours (group 1) or by 4 hours (group 2) of reperfusion ( P Conclusions Significantly elevated neuron-specific enolase serum levels could be demonstrated as a consequence of ischemia-induced cytoplasmic loss of neuron-specific enolase in central nervous system neurons, corresponded quantitatively to the severity of cerebral ischemia, and were detectable before irreversible neuronal injury. Therefore, analysis of serum neuron-specific enolase is suggested to be both a valuable diagnostic tool in clinical management of the initial stages of global cerebral ischemia and a prognostic parameter during the postischemic course.

We report on 7 cases of a nonnecrotizing type of autoimmune myopathy found in suspected connective tissue disease and related disorders. In muscle biopsy, these cases showed myopathic alterations characterized by irregular outlines of muscle fibers in cross and longitudinal section nearly or totally lacking inflammatory cell infiltrations or macrophages and no necroses of myofibers. They revealed deposition of immunoglobulins and of collagen IV antibodies attached to fiber surface. The number of capillaries per myofiber was enlarged. Capillaries of pipestem type with thickened wall were found at the electron microscopic level. This type of myopathy was seen especially in subacute cases presenting with muscle pain, weakness and increased unspecific inflammatory laboratory parameters. No evidence for polymyositis, dermatomyositis or inclusion body myositis could be found. In all cases, steroid therapy was of striking benefit.

Lipofuscin is a waste product of autolysosomal metabolism. The amount of lipofuscin in the cytoplasm depends on cell type, cell function and age. In most studies, either the fluorescent or the stained component of lipofuscin was investigated. Quantitative morphological investigations of the lipofuscin composition separated into the vacuolar and granular component were missing. For the hippocampal pyramidal cells we have determined the lipofuscin quantity and, separately, the vacuolar and granular component at the ultrastructural level. The hippocampal subfields CA 1, CA 2, CA 3 and CA 4 were observed at the ages 20, 40, 60 and 80 years (±3 years). Quantitative determinations of the vacuolar and granular component of neuronal lipofuscin in pyramidal cells were performed with a semi-automatic image analysis system. In CA 1 pyramidal cells the lipofuscin content was significantly lower than in the other sectors, which did not differ significantly in their lipofuscin content. The amount of the granular component in relation to the vacuolar component in CA 1 was larger than in the other sectors. With advancing age the lipofuscin content per cell increased. The vacuolar component of all hippocampal subfields experienced a larger increase than the granular component. Consequently the relation of the vacuolar and granular component changed; the relative amount of the vacuolar component increased, while that of the granular component decreased with age. The differences between sector CA 1 and the other hippocampal subfields were discussed with reference to differences of metabolic and functional activity of the neurons. Cytoprotective factors like Calbindin D28k were discussed for CA 1.

In Alzheimer's disease (AD), cortical neurons develop neurofibrillary tangles (NFTs) consisting of hyperphosphorylated tau. The neurons eventually die. There are some hints that cortical neurons may also degenerate without the development of cytoskeletal changes. We investigated this possibility by comparing changes in APP staining and neuronal size with respect to the presence or absence of hyperphosphorylated tau. Adjacent sections of the medial temporal neocortex (Brodmann's area 22) of 5 male AD patients aged 60-88 years (Braak V-VI) and 5 age-matched male non-demented control subjects were i) stained with a modified Bielschowsky silver method in order to reveal NFTs and 'ghost' tangles, ii) single-stained with anti-APP, and iii) double-labeled with anti-APP and AT8. Anti-APP is directed against the beta-amyloid precursor protein and stains virtually all perikarya and proximal neurites of the cortical neurons. AT8 stains pre-tangles, NFTs and extracellular 'ghost' tangles due to the recognition of hyperphosphorylated tau. The study was focused on the supragranular cortical layers II-III, since these layers can be clearly delineated from the adjacent molecular and granular cell layers. The results showed that i) APP staining intensity in neurons was variable in the AD cortex, being clearly different from the invariably intense neuronal staining in all controls. Reduced cytoplasmic APP staining was observed, particular in small neurons, while lack of anti-APP staining in proximal neurites, too, was associated with AD. In addition, ii) cross-sectional area measurement on anti-APP-stained neurons revealed that in AD, as compared to controls, a clear decrease in the number of mainly large-sized neurons (>150 microm2) was accompanied by a significant increase in the percentage of neurons in the smaller size classes, indicating that many large-sized neurons became smaller in AD. iii) Reduced APP staining and decreased neuronal size were not necessarily associated with the presence or absence of hyperphosphorylated tau in these cells. iv) Twenty-six percent of the neurons contained hyperphosphorylated tau, while the level of NFT-related neuronal loss was low in AD. The present study suggests that non-tau based neuronal degeneration is a major phenomenon in the AD neocortex.

To investigate a correlation between preoperative data from proton-MR-spectroscopy (1HMRS), genomic alterations (epidermal growth growth factor receptor [EGFR] gene amplification) and histomorphometric data from glioblastomas.In surgical specimens from 18 patients with glioblastomas, the degree of amplification of the gene for EGFR was determined in the region with the largest Ki-67 proliferation index by differential polymerase chain reaction.Correlation analysis showed significant positive correlation between degree of EGFR gene amplification and choline and total creatine (CHO/TCR) ratio, indicating increased membrane turnover. Cases with a high EGFR/interferon ratio showed a tendency toward a low lipid peak, whereas cases with a low EGFR/interferon ratio showed a large variation of the lipid peak. Differences were observed regarding quantitative histomorphologic data of tumor cell nuclei, especially nuclear size and shape. Together with the EGFR/interferon ratio, these morphometric data provided a good reclassification of cases with low and with high values for both spectroscopic variables by means of cross-validated linear discriminant analysis.The results provide further evidence for the biologic significance of metabolic data from preoperative 1HMRS, because these metabolic data showed a significant statistical relationship with histomorphology and a frequently occurring molecular biologic alteration (EGFR gene amplification) in glioblastomas.

A deeper knowledge about histopathological criteria with a significant impact on the prognosis of patients with glioblastomas is worthwhile, since these patients may show a considerable difference in the time of survival. Investigation of the morphology of perinecrotic tumor cell nuclei is a promising approach, because the expression of specific molecules in these cells has been associated with a more aggressive behaviour of the tumors. In our series of patients with documented clinical course, 11 patients had a survival of at least 24 months and we compared this group with a group of 10 patients with maximum survival of 12 months. Digital microscopic image analysis was performed in paraffin sections from the primary surgical specimen. Three hundred perinecrotic tumor cell nuclei per case and 300 nuclei per case from tumor cells lying more distant from the tumor necroses ('distant zone' nuclei) were measured. The ratio for the numerical nuclear density between both types of nuclei was significantly larger in cases with short survival indicating a more pronounced nuclear density of perinecrotic nuclei in relation to the 'distant zone' nuclei in these tumors. In cases with long survival, on the contrary, perinecrotic tumor cells exhibited an only slightly larger numerical density compared with 'distant zone' nuclei. Mean values and standard deviations from parameters of nuclear shape (Fourier-amplitudes) had significantly smaller values in short-time survivors indicating a tendency towards a more circular nuclear shape with less pronounced intratumoral variation in that group. Based on the morphometric results, all cases could be correctly reclassified as short- or long-time survivors by means of cross-validated discriminant analysis. In conclusion, the results confirm significant morphological differences between tumors from patients with short and with long survival regarding morphology of both types of tumor cell nuclei. It seems likely, that histomorphometry of tumor cell nuclei could be a promising approach for the assessment of the individual prognosis of patients with glioblastomas.

To study the discriminatory power of different methods designed for nuclear shape analysis with reference to the differentiation and grading of brain tumors and the differentiation between proliferating and nonproliferating nuclei.At least 300 tumor cell nuclei per case were measured by means of a digital image analysis system. Fourier amplitudes no. 1 to 15, moments no. 1 to 7 according to Hu, roundness factor, ellipse shape factor, concavity factor, Feret ratio, fractal dimension and bending energy were determined for each nucleus. The discriminatory power of these parameters was tested in three pairwise comparisons: (1) oligodendrogliomas WHO grade II (n = 13) vs. grade III (n = 11), (2) medulloblastomas WHO grade IV (n = 14) vs. anaplastic ependymomas WHO grade III (n = 12), (3) Ki-67-positive vs. Ki-67-negative tumor cell nuclei in the 14 medulloblastomas.When data from Fourier analysis were included in statistical analysis, cross-validated discriminant analysis led to a 100% correct reclassification for the first and for the second pairwise comparison and to a 75% correct reclassification when comparing Ki-67-positive and Ki-67-negative nucleifrom medulloblastomas. Different combinations of the other shape parameters led to a lower percentage of correctly reclassified cases for all three pairwise comparisons, especially when Fourier analysis was not included in the analysis.Fourier analysis provided an optimal statistical discrimination between different brain tumor entities and between data sets from proliferating and nonproliferating tumor cell nuclei. Since nuclear shape is an important criterion for the investigation of tumors, the application of Fourier analysis is therefore recommended for quantitative histologic investigations in neuro-oncology.

Although tumour cell nuclei are important histological structures for grading of astrocytomas according to the WHO-classification of brain tumours, there is no reported morphometric study of astrocytomas which describes quantitatively the four main morphologic criteria of tumour cell nuclei: size, shape, texture (densitometric characteristics) and spatial relationships between the nuclei (topometric analysis). Using a set of morphometric parameters describing these criteria as well as the Ki67-proliferation index, 74 astrocytomas from 74 patients were studied by means of a digital image analysis system. The objective of the study was to test, if these morphometric parameters were sufficient for statistical discrimination between pilocytic astrocytomas WHO-grade I, astrocytomas grade II and anaplastic astrocytomas grade III. Our results showed a correct reclassification of 97.3% (72/74) of the cases with respect to the tumour grade by means of cross-validated discriminant analysis. Morphometric parameters characterizing nuclear shape (shape factors, Fourier-amplitudes) showed the most prominent differences between the three groups of cases, followed by topometric parameters (number of neighbours per nucleus, distances between the nuclei). Less pronounced differences between the tumour grades were found for parameters characterizing nuclear size, nuclear texture and the Ki67-proliferation index. In conclusion, the present morphometric procedure provided good discrimination between the tumour grades, supporting the view that histomorphometry of tumour cell nuclei could be a valuable tool for grading of astrocytomas.

To study the regional heterogeneity of epidermal growth factor receptor (EGFR) gene amplification (EGFR-GA) in glioblastomas, considering the relationship between this mutation and morphology of tumor cell nuclei.Tissue samples gained by laser microdissection and pressure catapulting were used for the performance of differential polymerase chain reaction in 32 morphologically different regions from 7 glioblastomas. Semiquantitative determination of EGFR expression and image analysis of tumor cell nuclei were performed in the same regions.Distinct regional differences concerning the degree of EGFR-GA were found in 2 tumor cases. When comparing regions with different degrees of gene amplification within these cases, morphologic differences in tumor cell nuclei were observed. The other tumor cases also showed distinct intratumoral heterogeneity concerning histomorphology but no regional heterogeneity in the degree of EGFR-GA. When comparing regions with a low densitometric EGFR/interferon (INF) band ratio (2.19, n = 18) and a high EGFR/IFN band ratio (4.39, n = 14), the latter type of region showed a significantly higher percentage of Ki-67--positive tumor cell nuclei and lower values for several shape variables (Fourier amplitudes), indicating a tendency toward a more regular nuclear shape in regions with distinct EGFR-GA. For the EGFR/IFN band ratio, a significant correlation was found with several morphometric variables, especially those of nuclear shape and distances between nuclei.In glioblastomas showing regional heterogeneity in the degree of EGFR-GA, morphology of tumor cell nuclei has been shown to be different when comparing regions with different degrees of EGFR-GA. Glioblastomas may also show distinct regional heterogeneity of histomorphology without evidence of regional heterogeneity of EGFR-GA. A significant statistical association has been confirmed between the degree of EGFR-GA and quantitative morphology of tumor cell nuclei.

The technique of laser microdissection together with laser pressure catapulting (LMPC) is demonstrated in paraffin sections obtained from surgical specimens of brain tumors mounted on glass slides. A sufficient and precise application of microdissection techniques in tissue on glass slides is worthwhile, since it offers the possibility of a retrospective analysis of archived paraffin sections in histopathology. We could demonstrate a precise dissection of areas in tissues of different thicknesses (4 microm and 20 microm). Areas of tissue mounted directly on glass need to be dissected in a scanning mode in order to remove the total region in form of small tissue fragments row by row. This mode provided a precise microdissection of tissue areas of different sizes and shapes. A successful molecular biological analysis of the microdissected regions could be demonstrated. As an example for such an analysis, differential-PCR for detecting an amplification of the gene for the epidermal growth factor receptor (EGFR) was performed.

A comparison between data from proton-MR spectroscopy (1HMRS) and quantitative histomorphology of tumor cell nuclei in gliomas has not been reported up to now. Therefore, the question must be answered, if there are any significant correlations between histomorphology of gliomas and quantitative data from 1HMRS concerning tissue metabolites. Surgical glioma specimen (glioblastomas, astrocytomas, oligodendrogliomas) from 46 patients with tumor grades II-IV according to WHO have been evaluated by means of a digital image analysis system using Ki-67-immunostained paraffin sections. Nuclear density, Ki-67-proliferation index, nuclear area and shape variables (roundness factor, Fourier-amplitudes) have been determined from 200 randomly selected tumor cell nuclei in each tumor specimen. These data have been correlated with preoperative data from 1HMRS. A positive correlation between Fourier-amplitudes, choline peak and lipide peak was observed, as well as a negative correlation between these variables and the nuclear roundness factor. This result indicates higher choline and lipide peaks with increasing irregularity of nuclear outlines. Proliferation index Ki-67 was positively correlated with the lipide peak, nuclear density showed a positive correlation with the choline peak. Glioblastomas (n = 29) showed an additional positive correlation between mean nuclear size and total creatine. Anaplastic gliomas (n = 12) showed a positive correlation between lactate peak and the standard deviation of the nuclear roundness factor. Further multivariate analyses have shown, that for the present collective of 46 cases, histometric variables have a higher significance than spectroscopic data for the differentiation of the different tumor grades. These results verify a significant correlation between preoperative data from 1HMRS and histomorphology of tumor cell nuclei in gliomas, supporting the biological significance of both histomorphometry and 1HMRS for the evaluation of these tumors.

To investigate possible statistical correlations between metabolic data from preoperative proton magnetic resonance spectroscopy (1HMRS) and morphology of proliferating tumor cell nuclei in anaplastic gliomas and glioblastomas.Ki-67-positive tumor cell nuclei in paraffin sections of surgical specimens from 36 patients (7 anaplastic gliomas, World Health Organization grade 3; 29 glioblastomas, World Health Organization grade 4) were investigated by means of a digital image analysis system. Stringent inclusion criteria were formulated for all cases with respect to histologic quality and spectroscopic examination. As morphometric variables, nuclear area, shape variables (roundness factor, size-invariate Fourier amplitudes) and density of Ki-67-positive tumor cell nuclei per reference area were determined.Correlation analysis according to Spearman revealed a significant positive correlation between the total creatine (TCR) peak and nuclear area (P = .005). This correlation was also found within the glioblastoma group (P = .019). There was also a significant negative correlation of nuclear area with the ratio between choline and TCR in all cases (P = .014) and within the glioblastoma group (P = .046). No significant correlation of spectroscopic data was found with nuclear shape or density of Ki-67-positive tumor cell nuclei.The results demonstrate a correlation between spectroscopic data and morphology of proliferating tumor cell nuclei (nuclear size) in high grade gliomas. This study is part of a detailed investigation of the interrelationship between preoperative 1HMRS and quantitative histomorphology of gliomas.

To investigate differences between astrocytomas of WHO grade 2 and anaplastic astrocytomas of WHO grade 3 in terms of topometric variables characterizing individual tumor cell nuclei.Paraffin sections from surgical specimens from 25 astrocytomas (grade 2, n = 11; grade 3, n = 14) were analyzed by means of an image analysis system. At least 300 tumor cell nuclei were measured in the region with the highest Ki-67 proliferation index. Three different kinds of topometric variables were determined for each tumor cell nucleus: (1) several distances; (2) the variable Angle 2/1, the angle between the straight lines representing the distance to the nearest and second-nearest nucleus; and (3) the number of neighbors according to our mathematical definition.Most topometric variables showed distinct differences between the 2 tumor grades (multivariate analysis of variance), with 88% cases correctly reclassified by means of cross-validated discriminant analysis. The variables with the highest discriminatory power were the SD of Angle 2/1 and the ratio between the distance to the second-nearest and nearest tumor cell nucleus, with lower values for these variables in anaplastic astrocytomas. Even variables concerning neighborhood relationships showed significant differences.The results of this pilot study show that this first set of topometric variables is sufficient to detect differences between topologic characteristics of tumor cell nuclei in astrocytomas grade 2 and grade 3. Topometry seems to be an important tool for grading astrocytomas.

Brain tumor formation and growth is accompanied by the proliferation and infiltration of blood capillaries. The phenotypes of endothelial cells that make up capillaries are known to differ not only in the tissues in which endothelial cells are located but also as a result of the microenvironment to which they are exposed. For this reason, primary cultures of brain endothelial cells were isolated from human brain tumors removed by surgery and compared with cells from normal tissue. The primary confluent monolayers that grew out of isolated capillary fragments consisted of closely associated, elongated, fusiform-shaped cells. But brain tumor-derived endothelial cells in culture exhibited significantly less expression of endothelial-specific Factor VIII-related antigen compared with cells isolated from normal tissue. Cultured cells that exhibited binding of Ulex europaeus lectin were shown to take up Dil-Ac-Ldl and formed continuous monolayers that were joined together by tight junctions. The cells also exhibited characteristics of the cells of the brain microvasculature in vitro as seen by the presence of large numbers of mitochondria and few pinocytotic vesicles and by the absence of Weibel-Palade bodies within the cells. The expression of vascular cell adhesion molecule-1, E-Selectin, and the tight junction associated protein ZO-1 but not intercellular adhesion molecule-1 was demonstrated by immunohistological staining or reverse transcriptase-polymerase chain reaction methodologies. Comparative studies of these endothelial cells with endothelial cells from normal tissue will be useful for determining and understanding how the blood-brain barrier differs and functions in tumor and healthy tissues and may lead to strategies for brain tumor therapeutic approaches.

Postmortem neuropathologic reports for a consecutive series of 436 HIV-seropositive patients who died between 1985 and 1999 were matched with clinical data for 371 of them. Cases were divided into four groups depending on the date of death. The chosen time periods reflected the type of antiretroviral therapy available: before 1987 (before zidovudine); 1987-1992, the period of monotherapy (nucleoside analog reverse transcriptase inhibitors [NRTIs]); 1993-1995, the era of the use of dual NRTI combinations; and 1996-1999, the era of highly active antiretroviral therapy (HAART) containing protease inhibitors. Fifty-seven percent of our cases in this group had been prescribed HAART. In our study population, accessibility to the latest antiretroviral therapy was widespread. The total number of HIV autopsies declined after the advent of combination therapy. The prevalence of opportunistic infections-cytomegalovirus, toxoplasmosis, cryptococcosis, and central nervous system lymphoma-decreased over time. Cerebral tuberculosis, aspergillosis, herpes, and progressive multifocal leukoencephalopathy showed a downward trend, but the numbers were too low for statistical analyses. The incidence of HIV encephalopathy increased over time (p =.014). The rising prevalence of HIV encephalopathy at time of death may reflect a longer survival time after initial HIV infection in the HAART era. Although combination therapies decrease overall mortality and prevalence of CNS opportunistic infections, these therapies may be less active in preventing direct HIV-1 effects on the brain.

Proton MR spectroscopy ((1)H MRS)-visible total choline-containing compounds (tCho-compounds) are derivatives of membrane phospholipids and, in part, may act as a long-term second-messenger system for cellular proliferation. Experimental evidence suggests increasing concentrations of tCho-compounds during cellular proliferation. The present study was conducted in order to test the hypothesis that in vivo measurements of tCho-concentrations using (1)H MRS allow assessment of the proliferative activity of neuroepithelial brain tumors presurgically. Single-voxel (1)H MRS (PRESS, TR 1500 ms, TE 135 ms) was performed in 101 patients with neuroepithelial brain tumors prior to surgery and 19 healthy volunteers. Histological diagnoses were confirmed postsurgically according to the WHO classification. Measured tCho-compound signal intensities were corrected for coil loading, numbers of acquisitions and voxel size, and tCho concentrations calculated as institutional arbitrary units. They were matched with the mean immunohistochemical marker of cell proliferation, the Ki-67 (MIB.1) labeling index, using correlation analysis according to Spearman. Compared with low-grade tumors (i.e. WHO grade I/II) and normal white brain matter, high-grade tumors (i.e. WHO grade III/IV) revealed significantly (p0.05) elevated labeling indices paralleled by increasingly elevated tCho-concentrations. In contrast tCho-concentrations in low-grade tumor did not differ significantly from physiological values. A highly significant positive correlation (p0.0001, r(2) = 0.81) was found between the tCho-concentration and the labeling index. It was concluded that the determination of tCho-concentrations using in vivo (1)H MRS could provide a novel and noninvasive assessment of the proliferative activity of neuroepithelial brain tumors, pointing at (1)H MRS as a useful method for differentiating proliferating from non-proliferating tissues. Hence, potential indications for the clinical application of (1)H MRS are grading tumors presurgically, early detection of anaplastic transformation, and monitoring treatment.

The actin-binding protein ezrin has been associated with motility and invasive behavior of malignant cells. To assess the presence of this protein in human glial cells of the brain and its potential role in benign and malignant glial tumors, we studied ezrin immunoreactivity (IR), proliferation (MIB-1-IR), and apoptosis (terminal dUTP nick-end labeling) in normal human brain tissues from 10 autopsies and tissues from 115 cases of human glial tumors including astro-cytomas, ependymomas, oligodendrogliomas, and glioblastomas. We found weak staining of peripheral processes in normal human brain astrocytes and in World Health Organization grade II benign astrocytomas. Staining was markedly increased in anaplastic astrocytomas (World Health Organization grade III) and clearly strongest in glioblastomas (World Health Organization grade IV). The increase of ezrin-IR correlated significantly with increasing malignancy of astrocytic tumors (P < 0.0001). Statistical analysis revealed a stronger association with increasing malignancy for ezrin-IR than for MIB-1-IR or terminal dUTP nick-end labeling staining. Ezrin-IR was absent in normal oligodendrocytes and in oligodendrogliomas, but pronounced in normal ependymal cells and ependymomas. Ezrin-IR seems to be specific for astrocytes and ependymal glia in the normal brain. Our results indicate that ezrin-IR may provide a useful tool for the distinction of oligodendrogliomas and astrocytomas and for the grading of astrocytic tumors.

Language-relevant processing of auditory signals is lateralized and involves the posterior part of Brodmann area 22. We found that the functional lateralization in this area was accompanied by interhemispheric differences in the organization of the intrinsic microcircuitry. Neuronal tract tracing revealed a modular network of long-range intrinsic connections linking regularly spaced clusters of neurons. Although the cluster diameter was similar in both hemispheres, their spacing was about 20 percent larger in the left hemisphere. Assuming similar relations between functional and anatomical architecture as in visual cortex, the present data suggest that more functionally distinct columnar systems are included per surface unit in the left than in the right area 22.

Neuropathological diagnostic criteria for Creutzfeldt-Jakob disease (CJD) and other human transmissible spongiform encephalopathies (prion diseases) are proposed for the following disease entities: CJD--sporadic, iatrogenic (recognised risk) or familial (same disease in 1st degree relative): spongiform encephalopathy in cerebral and/or cerebellar cortex and/or subcortical grey matter; or encephalopathy with prion protein (PrP) immunoreactivity (plaque and/or diffuse synaptic and/or patchy/perivacuolar types). Gerstmann-Sträussler-Scheinker disease (GSS) (in family with dominantly inherited progressive ataxia and/or dementia): encephalo(myelo)pathy with multicentric PrP plaques. Familial fatal insomnia (FFI) (in member of a family with PRNP178 mutation): thalamic degeneration, variable spongiform change in cerebrum. Kuru (in the Fore population). Without PrP data, the crucial feature is the spongiform change accompanied by neuronal loss and gliosis. This spongiform change is characterised by diffuse or focally clustered small round or oval vacuoles in the neuropil of the deep cortical layers, cerebellar cortex or subcortical grey matter, which might become confluent. Spongiform change should not be confused with non-specific spongiosis. This includes status spongiosus ("spongiform state"), comprising irregular cavities in gliotic neuropil following extensive neuronal loss (including also lesions of "burnt-out" CJD), "spongy" changes in brain oedema and metabolic encephalopathies, and artefacts such as superficial cortical, perineuronal, or perivascular vacuolation; focal changes indistinguishable from spongiform change may occur in some cases of Alzheimer's and diffuse Lewy body diseases. Very rare cases might not be diagnosed by these criteria. Then confirmation must be sought by additional techniques such as PrP immunoblotting, preparations for electron microscopic examination of scrapie associated fibrils (SAF), molecular biologic studies, or experimental transmission.

The topographical distribution of adrenocorticotropic hormone, growth hormone and prolactin was studied in nine multihormonal pituitary adenomas and two normal adenohypophyses using quantitative immunohistochemical methods. For each pituitary hormone paraffin sections stained by antihormone antibodies were scanned with a light microscope equipped with a projection and scanning device. The number of immunoreactive cells of each square in the grid was counted and transformed into pseudocolors. In this way, the distribution of each hormone was documented. A chi-square test was applied to these distribution patterns to prove randomness. A nonrandom distribution of immunoreactive cells was observed in all pituitary adenomas and normal pituitary glands. Cluster formations of immunoreactive cells were found for each bormone. The clusters were surrounded by halo-like areas of decreasing number of immunoreactive cells from center to periphery. The cluster-halo type of immunoreactive cells was found not only in the adenomas but also in the normal pituitary gland. This may be explained by the similarity of biological processes during cell proliferation either in normal development or in adenoma formation. It can be concluded from our studies that pituitary adenomas in most cases develop by multicentric but not diffuse proliferation of cell clones of different endocrine activity.

The understanding of delayed hippocampal death as a therapeutic window for post-ischemic treatment of the brain has led to numerous investigations focusing upon underlying cellular mechanisms and pharmacological potentials in gerbils and rats. Nevertheless, studies on the occurrence of delayed neuronal death in the human brain have been singular and dealt with only small files of patients. To complement these limited data, in the present study 26 adult patients with a history of a single cardiac arrest were included. Following successful resuscitation, individual survival ranged from less than 1 h to 186 days (mean = 11 days). The severity of the resultant ischemic injury in hippocampus CA1, among Purkinje cells, or in frontal neocortex, respectively, was quantified by direct counting of necrotic neurons. Additionally, hippocampal specimens were immunostained for neuron-specific enolase. The data obtained demonstrate the occurrence of delayed neuronal death in human hippocampus and, in a minor form, in cerebellar Purkinje cells. This is in contrasts to the immediate manifestation of ischemic neuronal necrosis in the neocortex. Unlike previous findings in experimental animals and in humans, the delay of CA1 cell death could be defined as lasting about 7 days following cardiac arrest. Moreover, the immunohistochemical results indicate delayed neuronal recovery in CA1, which in the time course reciprocally corresponds to delayed manifestation of hippocampal neuronal death. Interpretation of the results must consider the lack of information about the exact individual duration of cardiac arrest and resuscitation, as well as missing data concerning pre-ischemic physiological variables.

Ein 64jahriger Patient aus unserer Klinik bemerkte im Januar 1990 zum ersten Mal eine Gehschwache, die sich bis zur stationaren Aufnahme in die Neurochirurgische Universitatsklinik in Frankfurt/Main im Juni 1990 zu einem inkompletten Cauda-Syndrom entwickelte. Aus der Vorgeschichte ist die operative Entfernung eines zervikalen, intraspinalextramedullaren Tumors im Jahre 1969 bekannt, dessen histologische Diagnose “Zylindrom” (adenoid-zystisches Karzinom genannt) lautete. Dieses Karzinom entsteht uberwiegend in den Mundspeicheldrusen (8). Ein Primartumornachweis gelang seinerzeit nicht, so das man keinen Hinweis fur ein metastatisches Geschehen hatte.

Focal and widespread intense hyperplasia of piloid astrocytes was found in the brain of a 51 year old woman with multiple sclerosis and typical demyelinating lesions. The piloid astrocytosis was confined to older plaques in brain stem, cerebellum and periventricular white matter, but could not be found in telencephalic plaques distant from the ventricular system. The piloid astrocytes and accompanying glial fiber bundles were oriented in alternating direction, thus imitating structural characteristics of spongioblastomas. In the pons, an especially high density of astrocytosis was found with lobulated and giant nuclei and binuclear cells. One large demyelinated plaque in the central white matter of the cerebellum contained numerous Rosenthal fibers between the piloid astrocytes. By their intense glial fiber production, the piloid astrocytes took part of the formation of glial scars. The term spongiocyte is proposed for piloid astrocytes of this highly differentiated type, in order to separate them from the immature spongioblasts of gliogenesis. The pathogenetic mechanism is not clear. It is supposed that during intense reparative astrocytic proliferation the development of atypic reactions is favoured. A primary evocation of this type of reaction by the demyelinating factor itself must also be taken into consideration. The topographic predilection, in our case, of piloid astrocytosis to brain stem and cerebellum indicates a disposition of regions formed by the embryogenetic side plate to reactions of this type.

Befunde von drei Patienten aus einer Familie mit einem Morbus Gerstmann-Straussler-Scheinker (M-GSS) werden mitgeteilt. Die Diagnose wurde zweimal autoptisch gesichert. Klinisch und morphologisch handelt es sich um eine seltene familiare subakute spongiose Encephalopathie mit besonderem Befall des Kleinhirns, deren Vorkommen bisher nur in vier Familien mit insgesamt 52 Kranken gesichert ist. In der hier untersuchten Familie begann die Erkrankung durchschnittlich im funften Dezennium und fuhrte innerhalb von funf Jahren zum Tode. Cerebellare Funktionsstorungen, spater Demenz, bulbare und Pyramidenbahn-Symptome sowie Hor- und Sehminderung bei zwei Patienten, charakterisierten das klinische Bild. Die biochemischen Befunde, einschlieslich deren des Liquors, waren unauffallig. Neurophysiologisch zeigte sich mit fortschreitender Erkrankung eine Allgemeinveranderung im EEG ohne periodische paroxysmale Ablaufe. Fur eine Demyelinisierung ergaben die evozierten Potentiale keinen Hinweis. Nur durch die histologischen Befunde mit den nicht immer nachweisbaren Kuru-Plaques, aber stets unterschiedlich ausgepragten multizentrischen Plaques, spongiosen Ver↭derungen wechselnder Auspragung war die Erkrankung sicher vom M. Alzheimer und M. Creutzfeldt-Jakob (M-CJ) abzugrenzen. Nach ersten positiven Ubertragungsversuchen auf Primaten und ahnlichen histologischen Veranderungen beim M-CJ und Kuru wird eine slow-virus-Infektion durch ein nichtkonventionelles, moglicherweise dem M-CJ-, Kuru- und Scrapie-Virus verwandtes Virus diskutiert. Die unterschiedliche Manifestation der beim Menschen beobachteten Erkrankungen konnte auch durch verschiedene dispositionelle Faktoren beim Wirt erklart werden, die nach den bisherigen humangenetischen Befunden beim M-GSS autosomal dominant weitergegeben werden.

Light and electron microscopic observations are reported on a brain biopsy of a man of 59 with a rare familial disease of the CNS and a 5-year clinical course. Electron micrographs of the frontal biopsy reveal plaque-like deposits composed of amyloid cores, often multicentric in the cortex and subcortical white matter. They are localized between enlarged astrocytic processes. In the neuropil they are sometimes associated with abnormal neuritic processes, in the white matter with processes of fibrous astroglia and basement membranes. There are no signs of primary neuritic or synaptic involvement in the plaque formation which is more obviously associated with altered astrocytic processes. Moreover, degenerative alterations in the cortical vessels and slight astroglial spongiform changes as well as oligodendroglial proliferation can be found. Plaques are considered to be mainly of the kuru type; the relationship with transmissible spongiform encephalopathies is discussed.

Der N. opticus von erwachsenen weissen Ratten wurde 5 mm retrobulbar scharf durchtrennt. Nach 24–48 Std. findet man im Axoplasma aufgetriebener Faserstumpfe distal der Lasion vesico-tubulare endoplasmatische Reticula, Filamente, Anhaufungen von Mitochondrien und dense bodies” sowie Schichtenkorper, die aus schalenartig gewolbten Doppellamellen bestehen und die im Axoplasma der Opticusfasern normalerweise nicht vorkommen. Die Lamellen entstehen durch Fusion sackartig sich ausweitender Anteile des endoplasmatischen Reticulum. Bei der Anreicherung der Mitochondrien spielen wahrscheinlich stumpfwarts gerichtete Axoplasmabewegungen ursachlich eine Rolle; daneben bestehen Anhaltspunkte fur eine abschnittweise Querteilung der Organellen. Die dense bodies” reprasentieren besondere Zustandsformen der Mitochondrien, die wahrend reaktiver Vorgange in neuronalem Cytoplasma vorkommen. Die Mitochondrien neigen zu schalenformiger Apposition aneinander unter Verdunnung und Abflachung ihrer Mittelteile und verschiedenartiger Transformation der Cristae. Schichtenkorper konnen durch Apposition derartig sich umwandelnder Mitochondrien an Umfang zunehmen. Wahrend der Formanderungen bieten die Mitochondrien keine Anzeichen einer Nekrobiose. Im Anschluss an diese primaren, reaktiven Vorgange setzen nach dem 2. Tag im Axoplasma regressive Veranderungen ein, die den Beginn der sekundaren Degeneration kennzeichnen.

Die drusige Entartung der Arterien der Hirnrinde im Senium wird an einem besonders ausgepragten Fall eines im 65. Lebensjahr unter dem Bild der senilen Demenz Verstorbenen polarisationsoptisch und elektronenmikroskopisch untersucht. Die Resultate der beiden Untersuchungsmethoden erganzen sich gegenseitig. In den degenerierenden kongophilen Gefaswanden sind bei voller Ausbildung des Prozesses polarisationsoptisch vier Schichten unterscheidbar, in denen abwechelnd senkrecht zueinander orientierte anisodiametrische Teilchen vorliegen; ihnen entsprechen elektronenmikroskopisch in gleicher Weise angeordnete 100 A breite Filamente. Sie sind in der Tunica media parallel zur Gefaslangsachse sowie zirkular ausgerichtet, in einer inneren und auseren, lichtmikroskopisch gestreiften Schicht sind sie senkrecht zur Gefasachse — radiar zum Lumen —, in einer zwischen diesen gelegenen Zone wieder parallel zur Gefasachse orientiert. In den wesentlichen tinktoriellen Resultaten, in den polarisationsoptischen und feinstrukturellen Befunden stimmen die entarteten Hirngefasewande im Senium mit den Ablagerungen in den Korperorganen bei allgemeiner Amyloidose uberein; das spricht fur ihre Amyloidnatur. Die vonScholz gewahlte Bezeichnung “drusige Entartung der Hirnarterien” wird dem feinstrukturellen Bild gerecht; gemeinsames Charakteristicum der gestreiften Schichten der kongophilen Gefaswand und der “Kerne” der senilen Drusen ist die keiner ortsstandigen Struktur folgende Formierung der filamentaren Amyloidkomponente in parallel oder radiar geordneten Bundeln, also die eigengesetzliche, kristallartige Ausfallung an diesen Orten. Strauchartig uber die Gefaswand hinaus sich erstreckende Filamentbundel entsprechen dem Ubertritt kongophiler Substanz in das Gewebe. Ein Kohlenhydratbestandteil des Amyloid ist wahrscheinlich an die filamentare, die Eiweiskomponente gebunden. Uber die zweite, amorphe Komponente des Amyloid lassen sich keine Angaben machen. Die Beziehung der drusigen Gefasentartung zu den unter dem Terminus “kongophile Angiopathie” beschriebenen Wandveranderungen und die Verwendung dieses Terminus werden diskutiert.

Nach Durchtrennung der Hinterstrange am Ruckenmark der weisen Ratte kommt es peritramatisch in den Axonstumpfen zu einem strukturellen Umbau im Cytoplasma der Axone bei meist erheblicher lokaler Vergroserung des Axondurchmessers (Axonauftreibungen). Im Axoplasma, das normalerwiese verhaltnismasig arm an geformten Strukturelementen ist, treten Zisternensysteme unterschiedlicher Auspragung und Dichte (endoplasmatische Reticula), Mitochondrien verschiedener Form und Grose in betrachtlicher Anzahl, Vesikel in wechselnder Verteilung sowie filamentoses Material auf. Die Veranderungen sind 12 Std nach der Durchschneidung bereits deutlich ausgepragt, der Hohepunkt wird zwischen dem 3. und 7. Tag erreicht. Die Beobachtungen lassen den Schlus zu, das die vorgefundenen Strukturen und Organellen vorwiegend an Ort und Stelle entstehen, sie sind aus einer Anstauung oder Wanderung praexistenter Elemente nicht allein erklarbar. Die als Reaktion auf die exogene Schadigung rasch in Gang kommenden anabolen Vorgange in den lasionsnahen Axonabschnitten, die der “primaren retrograden Reaktion” vonSpatz entsprechen, stellen den Versuch des Neurons dar, von hier aus den verlorengegangenen distalen Anteil des Zellfortsatzes wieder aufzubauen. In den Axonauftreibungen treten jedoch nach wenigen Tagen regressive Veranderungen auf, die sich in einer Dichtezunahme im Grundcytoplasma und in der Matrix der Mitochondrien zeigen und bis zur Aggregation der vorher aufgebauten Strukturen fortschreiten. Die Axonauftreibungen gewinnen schlieslich Fremdkorpercharakter und konnen, wie aus lichtmikroskopischen Untersuchungen hervorgeht, monate- bis jahrelang im Gewebe persistieren. Isolierte Axonkugeln sind vermoge ihres Mitochondrienreichtums in der Lage, uber mehrere Tage einen gewissen Stoffwechsel aufrechtzuerhalten, bevor sie der Nekrobiose verfallen. Anzeichen fur eine Bildung junger Axone innerhalb der Axonauftreibungen waren nachweisbar.

Verschiedenheiten in der Verteilung der 3 Gliazelltypen Astroglia, Oligodendroglia und Mikroglia in den einzelnen Schichten verschiedener nervenzellarchitektonischer Felder der Groshirnrinde wurden erstmals von Schroeder festgestellt, und zwar durch zeichnerische Wiedergabe der Zellen nach Nissl-Praparaten und nach Anwendung der Metallimpragnationsverfahren. Die sichere Erfassung architektonischer Verschiedenheiten der Neuroglia ist jedoch nur durch Zellzahlungen moglich; die Zahlung mus wegen der Unzuverlassigkeit der Metallimpragnationsverfahren an Nissl-Praparaten erfolgen. In der vorliegenden Arbeit wurden unter Zugrundelegung der 5 Rindentypen von v. Economo an 8 verschiedenen nervenzellarchitektonischen Feldern (Areae) der normalen menschlichen Groshirnrinde Zahlungen jedes der 3 Gliazelltypen getrennt in jeder Rindenschicht (Laminae) durchgefuhrt. Die Methodik der Schnittherstellung und der Zahlung und die morphologische Unterscheidung der 3 Gliazelltypen im Nissl-Bild werden beschrieben. Nach den erhaltenen Zahlenwerten wurden Kurven aufgestellt. Die Ergebnisse sind folgende: Ruckschlusse auf die normale Funktion der Gliazellen sind allein auf Grund der festgestellten zahlenmasigen Beziehungen zwischen ihnen und den ubrigen Gewebselementen nicht moglich. Nach Bestatigung an einem groseren Material und in Zusammenhang mit Ergebnissen aus anderen Forschungsrichtungen konnten diese Beziehungen jedoch wesentliche Bedeutung haben. Fur die Erfassung des Verhaltens des gliosen Interstitiums unter pathologischen Bedingungen ist der Nachweis des Vorhandenseins einer laminar und areal differenten Gliazellarchitektonik in der Hirnrinde von Wichtigkeit. In den gewonnenen Zahlen stehen hiermit Vergleichswerte zur Verfugung.

Ein 60 jahriger Landwirt erkrankte aus voller Gesundheit 4 Monate vor dem Tode mit Kopfschmerzen, Schwindelerscheinungen und Angstzustanden. Unter dem Verdacht eines raumfordernden Prozesses erfolgte Krankenhausaufnahme. Neurologisch Koordinationsstorungen, corticale Sehstorungen, rasche Progredienz der Symptomatik, zuletzt Rindenblindheit, Bewustseinsveranderungen, epileptiforme Krampfanfalle. Im EEG Verlangsamung, steile Wellen mit trager Vor- und Nachschwankung, kein Herdbefund. Histologische Untersuchung des Gehirns ergab verbreitet neuronale Zellausfalle, atrophische und geschrumpfte Nervenzellen in der Groshirnrinde mit fronto-occipital zunehmender Intensitat und schwerster Auspragung in der Area striata. In den neuronal geschadigten Rindenabschnitten Hyperplasie groskerniger, plasmareicher Astrocyten mit Gliarasenbildung in der Area striata. In anderen Rindenabschnitten disseminierter oder laminarer bis totaler gliogener (astrocytarer) Status spongiosus ohne Nervenzellschaden und ohne Gliavermehrung. Die beiden Schadensmuster wechseln meist miteinander ab, Uberlagerung kommt seltener vor. Kleinhirn, Hirnstamm und Halsmark sind unauffallig. Wahrend die neuronalen Schaden mit den klinisch-neurologischen Herdzeichen korrespondieren, wird der gliogene Status spongiosus, der eine unregelmasige Verteilung zeigt, zu den EEG-Veranderungen und zu dem dyskinetischmyoklonischen Endstadium der Erkrankung in Beziehung gesetzt. Fehlende Capillarfullung in Rindenabschnitten mit schwerer spongioser Dystrophie last auf sekundare Storungen der Mikrozirkulation durch generalisierten Astrocytenhydrops schliesen.

1. Das Endothelcytoplasma der Tumorgefase von Gliomen — Spongioblastomen, Astrocytomen, Oligodendrogliomen, Ependymomen und Glioblastomen — enthalt oft reichlich filamentares Material. Die Filamente gleichen den Filamenten im Cytoplasma der Geschwulstzellen selbst sowie den Filamenten der normalen und reaktiven Astroglia (100 A Durchmesser, kompakter Bau, unterschiedliche Langenausdehnung, unverzweigt). Die Bedeutung derFilamente im Endothel der Gliomgefase ist noch unklar. Im Gegensatz zu den Gefasen der Korperorgane treten im ZNS, soweit bisher bekannt, weder normalerweise noch im Rahmen mesenchymaler Reaktionen filamentare Elemente im Gefasendothel auf. 2. In einem Spongioblastom wurde ein disseminiert-herdformiger Zerfall der Gliafilamente beobachtet, der mit einer Anhaufung osmiophilen Materials an den zerfallenden Filamenten und im interfilamentaren Grundcytoplasma der betroffenen Bezirke einhergeht. Der Prozes spielt sich in den filamentreichen Zellfortsatzen der Spongioblasten ab, seltener in ihren Pericarya. Durch Konfluenz der Herde entstehen ausgedehnte langliche osmiophile Korper von granularer bis amorpher Feinstruktur und hoher Dichte, die die Fortsatze ausfullen und oft auftreiben. Sie entsprechen den sogenanntenRosenthalschen Fasern der Spongioblastome. Die Gebilde werden als Ergebnis einerStapelungsdystrophie der Spongioblasten angesehen. Gestapelt werden Proteine, fakultativ Kohlenhydrate, in spaten Stadien auch Lipide. Die Rosenthalschen “Fasern” gehen also aus faserhaltigen Zellteilen hervor, sind aber selbst keine Fasern.

In Achsenzylinderauftreibungen, wie sie in grauer und weiser Substanz des menschlichen Gehirns nach exogenen Schadingungen (Trauma, kreislaufbedingte Erweichungen) vorkommen, wurden mit dem Phasenkontrastverfahren darstellbare saurefuchsinophile und eisenhamatoxylin-farbbare Substanzanhaufungen festgestellt, die sich nach Anwendung histochemischer Methoden als glykolipoid-und proteidhaltig erwiesen. Es wird angenommen das sie durch Aufbau mitochondrienreicher cytoplasmatischer Strukturen in den aufgetriebenen Axonabschnitten zustande kommen, das es sich also hier nicht, wie bisher beschrieben, vorwiegend um Flussigkeitsaufnahme (Quellung) handelt. Die so gekennzeichneten Achsencylinderveranderungen sind Ausdruck der am proximalen Stumpf ladierten Nervenfasern ablaufenden “primaren retrograden Reaktion”. Sie stellen den Versuch der Nervenzelle dar, durch Intensivierung des Stoffwechsels und Aufbau neuer Cytoplasma bestandteile in weiter Entfernung vom Perikaryon den verlorengegangenen peripheren Anteil des Axons von der Lasionsstelle aus wieder aufzubauen. Am Inhalt der Axonauftreibungen kommt es jedoch schon nach kurzer Zeit zu Stoffumwandlungen, es laufen regressive Veranderungen ab; infolge des entstandenen fremdkorperartigen Charakters konnen isolierte Axonauftreibungen dann jahrelang unabgeraumt im nervosen Gewebe liegenbleiben.

9 Std nach experimentellen traumatischen Lasionen am Ruckenmark der Ratte wurden peritraumatisch in der grauen Substanz der Hinterhorner die Folgen einer Plasmaexsudation in das Gewebe elektronenmikroskopisch beobachtet. Es kommt zu plasmatischer Durchtrankung des Extracellularfugensystems bei Erhaltung der dicht gefugten Lagerung der Zellteile im Neuropil. Es folgen Einrisse der Zellmembranen und Eintritt von Plasmabestandteilen in die Zellfortsatze. Im Cytoplasma akkumulieren Blutplasmabestandteile in rundlichen membranumgrenzten Korpern, die aus dem endoplasmatischen Reticulum stammen. Fibrin wird in Form fein verteilter Faden eingelagert. Zwischen Gefaswanden und Neuropil finden sich Fibrinaggregate mit typischer Querstreifung. Die Zellfortsatze zerfallen schlieslich, in den lockeren Nekrosebezirken restieren dichte Korper und Doppelmembranbruchstucke. Die plasmatische Infiltrationsnekrose (Scholz) unterscheidet sich somit nicht nur pathogenetisch, sondern auch feinstrukturell von den primar hypoxischen Gewebsschaden (Kolliquations- und Koagulationsnekrose). In der Umgebung der Nekrosezonen tritt ein kollaterales Odem mit Schwellung der Zellfortsatze auf. Aus den Befunden geht in ubereinstimmung mit Beobachtungen anderer Autoren hervor, das die extracellularen Fugen im ZNS in vivo existieren und von einem leicht permeablen Medium erfullt sein mussen. Sie erlauben nicht nur die Verteilung ionaler Losungen, sondern auch kolloidal geloster Stoffe in der Grosenordnung der Plasmaproteine.

Inhaltsubersicht: Aktueller Stand von Epidemiologie und Ubertragungsmodus der HIV-Infektion.- Das humane Immundefizienzvirus HIV: Struktur und Pathogenese.- Biologisch unterscheidbare Subtypen von HIV in Blut und Liquor cerebrospinalis von AIDS- und LAS-Patienten mit neurologischer Symptomatik.- Nachweis von immunoglobulinklassen- und subklassenspezifischen Antikorpern gegen HIV.- Neurologische Probleme bei AIDS.- Zentralnervose Befunde bei 140 Frankfurter Patienten mit HIV-Infektion.- CT-Befunde am Gehirn bei AIDS.- Peripheres Nervensystem und AIDS.- Neuropathologische Beobachtungen in 28 Fallen von erworbenem Immundefektsyndrom (AIDS).- Das morphologische Korrelat der HIV-Infektion des Gehirns.- Allgemeine Pathologie bei AIDS und ihre Beziehungen zu neuropathologischen Befunden.