Mitogenic lectin receptors of nervous system tumors. Study of gliomas, neural crest tumors, and meningiomas in vitro using phytohemagglutinin and concanavalin A.

The effects of mitogenic lectins Phytohemagglutinin (PHA), and Concanavalin A (Con A) on the growth rate of cells derived from glial tumors (astrocytoma, ependymoma, glioblastoma, medulloblastoma, and C6 rat glioma), neural crest tumors (neuroblastoma and schwannoma), and meningiomas were studied. The cell lines were of human and animal origin. The specificity of lectin binding to mitogenic receptors was evaluated using complementary monosaccharides. In all glial- and some neural-crest tumor-derived cell lines, there was a lectin concentration-dependent and cell density-dependent, biphasic growth rate response with stimulation at low and inhibition at high lectin concentrations. This response did not depend on the type of glial tumor, species of origin, or passage level in vitro. Although, in meningioma-derived cell lines, lectins did not induce a growth rate response, they caused morphological changes ("whorling"). Lectin stimulation in glial tumor-derived cell lines resembles that occurring in peripheral blood lymphocytes. Lectin-induced mitogenesis may lay the groundwork for the establishment of a model of glial cell proliferation, and that permits the evaluation of cell surface effects, intracellular mechanisms, and epigenetic factors in studies of tumors, neural development, and neuroimmunology.

Hypomelanosis of Ito (incontinentia pigmenti achromians)--a clinicopathologic study: macrocephaly and gray matter heterotopias.

We studied a boy with macrocephaly, hypotonia, pigmentary retinopathy, unilateral whorled hypopigmented skin lesions, and seizures. Skin biopsy confirmed the clinical diagnosis of hypomelanosis of Ito. Postmortem examination at age 22 months revealed a severe neuronal migrational defect that altered the cerebral cortex architecture of white matter. There were many gray matter heterotopias characterized by altered neurons and giant cells. Electronmicroscopy revealed the astrocytic nature of the giant cells. Embryologic migration of both melanoblasts from neural crest and cortical neurons occurs in the second trimester, suggesting a common mechanism for the developmental pathology of skin and brain.

Ornithine decarboxylase activity in vitro in response to acute hypoxia: a novel use of newborn rat brain slices.

In fetal as well as newborn rats, acute hypoxic exposure results in significantly elevated brain ornithine decarboxylase (ODC) activity, polyamine concentrations, and ODC mRNA. The interpretations of these in vivo hypoxic-induced changes, however, are complicated by maternal confounding effects. To test the hypothesis that acute hypoxia will also increase ODC activity in vitro, we developed a brain slice preparation which eliminates such maternal effects. Sections of whole cerebrum, approximately 300-500 microns thick, were made from 3- to 4-day old Sprague-Dawley rat pups. The slices were equilibrated for 1 h in artificial cerebrospinal fluid (ACSF) continuously bubbled with 95% O2/5% CO2, prior to induction of hypoxia. We induced hypoxia by changing the oxygen concentration to 40%, 30%, 21%, 15%, 10%, or 0% O2, all with 5% CO2 and balance N2. In the normoxic control brain slices, low but stable basal ODC activity persisted for up to 5 h post-sacrifice. Slices in ACSF treated with bovine serum albumin (BSA), or both BSA and fetal bovine serum (FBS), however, showed stable ODC activity values 2- to 3-fold higher than slices in ACSF alone, for up to 5 h. In response to acute hypoxia (i.e., 15, 21, and 30% O2), ODC activity was elevated 1.5- to 2-fold above control values between 1 and 2 h after initiation of hypoxia. Qualitative light and electron microscopic examination of the neonatal brain slices following 2 h hypoxic exposure suggested that the great majority of cells did not show severe hypoxic damage or necrosis. It was concluded that: (1) in neonatal rat brain slices in vitro, stable ODC activity values approximating the whole brain ODC activity seen at sacrifice, can be maintained for several hours; (2) the in vivo hypoxic-induced increase in ODC activity can be approximated in vitro; (3) the neonatal rat brain slice preparation may be an alternative to other methods for studying hypoxic-induced ODC enzyme kinetics, or other brain enzymes, without maternal confounding effects; and (4) ODC activity may be an indicator of active metabolism within the newborn brain slice both in normoxia and hypoxia.

Electrophysiological and biochemical characterization of a continuous human astrocytoma cell line with many properties of well-differentiated astrocytes.

Astrocytes comprise about 25% of the cellular volume of the brain, and their main function is to maintain homeostasis of the neuronal environment. These cells are commonly identified on the basis of their membrane electrical properties and the presence of specific proteins. We have characterized the human astrocytoma cell line designated UC-11MG and have shown these cells have many of the traits of differentiated astrocytes. Many of the UC-11MG cells have a large resting membrane potential, averaging -74 mV. The slope of the Em vs log [K]o cuve was 58.5 mV per decade [K]o. The cells were inexcitable when exposed to brief depolarizing current pulses. The astrocytoma traits are virtually identical to those previously reported for normal astrocytes. The astrocytoma cells also express glutamine synthetase activity which is considered specific to astrocytes among brain cells. Previous work had also demonstrated the presence of other astrocyte markers glial fibrillary acidic protein and S-100 protein in the UC-11MG cells. The steady-state ion transport properties of Na+, Cl-, and K+ were also characterized in these cells, and the rates of efflux were found to be similar to those in other astrocytes, with the major difference being the presence of a second kinetic compartment in the UC-11MG cells. From this work, we conclude that the UC-11MG cell line displays prominent features associated with differentiated astrocytes, and may provide an excellent model system for the study of human astrocytes.

Pericyte degeneration and thickening of basement membranes of cerebral microvessels in complex partial seizures: electron microscopic study of surgically removed tissue.

Complex partial seizures are associated with alterations in regional cerebral blood flow in abnormally spiking foci, as shown by positron emission tomography and single photon emission computed tomography, with an increase in flow ictally and a decrease interictally. Alterations of vasoregulation during ictal periods have also been described in animal seizure models. An electron microscopic study on human brain tissue from seven patients undergoing resections for the treatment of intractable complex partial seizures was performed to examine ultrastructural changes of the microvasculature and their locations within the microvessel wall. Biopsies were obtained intraoperatively from temporal lobe regions with electrocorticographically detected abnormal spiking and from regions without abnormality on electrocorticograms (control samples) removed as part of the therapeutic resection. A total of 539 microvessels from three regions were evaluated: spiking mesial temporal lobe, spiking lateral temporal cortex, and nonspiking (control) cortex. Evidence of pericyte degeneration (aggregates of cellular debris within the basement membrane) was noted in the majority of spiking area microvessels (76.7% in spiking mesial temporal cortex; 69.8% in spiking lateral temporal cortex) as compared with 37.8% of control microvessels (P less than 0.0005). Morphometric studies revealed a significant increase in total wall thickness, pericyte-basement membrane unit thickness, pericyte cytoplasmic density, basement membrane density, and basement membrane thickness in microvessels from spiking (mesial and lateral temporal cortex), as compared to control areas (P less than 0.01). No statistically significant difference was noted in pericyte coverage or pericyte or endothelial mitochondrial densities between microvessels in spiking and control regions. This study shows degeneration of pericytes, cells thought to play an essential role in microvascular hemodynamics, and thickening of microvessel walls in abnormally spiking brain regions in patients with intractable complex partial seizures. The pericyte degeneration and basement membrane thickening in abnormally spiking areas may explain alterations in vasoregulation, by a decrease in the microvascular compliance and in cross-capillary diffusion.

Radiation-associated gliomas: a report of four cases and analysis of postradiation tumors of the central nervous system.

Four cases of radiation-associated gliomas are described. All patients were white men, irradiated in childhood for craniopharyngioma, anaplastic ependymoma, retinoblastoma of the orbit, and Burkitt's lymphoma, respectively. The dose ranged from 1800 to 5900 rads, and the latency period was 5 to 25 years. All primary and secondary tumors were verified histologically, and no evidence of persistence of the primary tumors was found. All secondary tumors arose in the fields of irradiation. Ninety-six cases of radiation-induced tumors of the central nervous system have been reported in the literature to date. Twenty-four were gliomas and occurred mainly in young men.

Giant granular cell tumor of the suprasellar area: immunocytochemical and electron microscopic studies.

We describe a case of a granular cell tumor (GCT) of the suprasellar region with an 11-year history in a 26-year-old woman. The computed tomographic scan showed a midline, contrast-enhancing, noncalcified mass. The biopsy was diagnosed as GCT. The tumor was treated with radiation therapy. At necropsy, a large, homogeneous GCT surrounded by gliosis was found. The tumor cells were filled with granules positive for periodic acid-Schiff, diastase-resistant. The cells did not contain glial fibrillary acidic protein or S-100 protein. Electron microscopy showed tumor cells filled with innumerable lysosomal structures. No intermediate filament was found within the cytoplasm. The tumor cells were not surrounded by a basement membrane. Based on this study and on our review of the literature, the suggestion that GCT has a multicellular origin is upheld.

Adrenoleukodystrophy: white matter disease with adrenal changes in 33-year-old man.

Patients with changes such as those that occur in relatively diffuse disorders of higher nervous function constitute a major diagnostic problem. We are presenting a patient with dysphasia, incoordination, difficulty concentrating, and weight loss with underlying organic changes. This 33-year-old patient had adrenoleukodystrophy (ALD), which is a sex-linked recessive disorder involving the adrenal glands with central nervous system demyelination. ALD usually manifests itself during childhood.

Alzheimer's disease underlies some cases of complex partial status epilepticus.

Alzheimer's disease is a known risk factor for seizures, and age older than 60 years is a recognized risk factor for poor outcome from convulsive and nonconvulsive status epilepticus. The authors suspect that there may be a causal relationship between dementia pathology and the development and maintenance of refractory seizures. They report two selected patients with complex partial status epilepticus whose presentation and clinical course provide partial support for this hypothesis. Their methods include case reports with clinical, EEG, imaging, and pathologic correlations. The patients were 70 and 85 years of age. Both had central and peripheral brain atrophy on imaging studies (with some regions that were affected more than others), left temporal seizure foci corresponding to areas of greatest cortical atrophy, and early presentation with inhibitory epileptic symptoms (aphasia), with evolution to complex partial status epilepticus. Pathologic confirmation of Alzheimer's disease was obtained in one patient who had not been diagnosed previously. It involved maximally the cortex underlying the seizure focus. A diagnosis of probable Alzheimer's disease was established in the other patient. Alzheimer's disease may be causal in some cases of complex partial status epilepticus. Additional observations in support of this hypothesis are needed.

Bilateral trigeminal neurofibrosarcoma. Case report.

An autopsied case of bilateral trigeminal neurofibrosarcoma is reported. The right-sided inferior alveolar tumor was treated surgically and subsequently irradiated. There was no local recurrence during the ensuing 4 years. Two years after excision of that tumor, a left-sided trigeminal neurofibrosarcoma was subtotally removed. Two years later this same tumor was found to have extensively invaded the pons.

Giant pituitary cavernous hemangioma: case report.

A large cavernous hemangioma of the pituitary was an incidental finding at the autopsy of a 72-year-old woman who died as a result of metastatic breast carbinoma. This lesion did not cause any overt clinical problems.

The relationship between the capillary structure and hemorrhage in gliomas.

A 48-year-old man was admitted with the sudden onset of symptoms of stroke caused by hemorrhage in an oligodendroglioma. Despite surgery and antiedema treatment, the patient died. Histological evaluation revealed an oligodendroglioma with calcified capillaries of the retiform type. To further investigate this phenomenon, a total of 160 gliomas were reviewed: 90 glioblastomas multiforme, 30 oligodendrogliomas, and 40 astrocytomas. Sufficient data were available for clinical evaluation in 100 cases. Of these, 5% (two oligodendrogliomas and three glioblastomas multiforme) were related to clinically significant hemorrhages. Of the remaining cases, microhemorrhages were found in 53.0% of the glioblastomas, in 56.7% of the oligodendrogliomas, and in 10.0% of the astrocytomas. In each case reviewed, the capillaries were assigned to one of three groups: axial, retiform, or glomeruloid. Statistical analysis revealed a significant association between hemorrhages and retiform capillaries in all three types of tumors, except that in oligodendrogliomas the statistical significance held true when calcification of the capillaries was also present. Glomeruloid-type capillaries were only weakly associated with hemorrhages, and no association was found for axial capillaries. A large-scale prospective study is necessary to more precisely assess the role of each of the three types of capillaries in hemorrhages of gliomas. Based on data available so far, patients with glial tumors with retiform capillaries, confirmed on biopsy, should be carefully monitored to exclude possible intratumoral hemorrhage.

Acute responses to blunt head trauma. Experimental model and gross pathology.

This study of blunt craniocerebral trauma describes an experimental model that involves delivery of forceful blows to the resting movable skulls of anesthetized cats. Injuries inflicted by this method included skull fractures in 81% of cases, epidural hemorrhages in 50%, subdural hemorrhages in 80%, subarachnoid hemorrhages in 100%, and brain contusions in 84%. In the majority of instances the subdural and epidural hemorrhages were thin films of blood that did not compress or distort the subjacent brain. The distribution of cerebral contusions was restricted to the cerebral parenchyma beneath the locus of cranial impact except for contusions associated with skull fractures. This experimental model recapitulates clinically realistic human cranial trauma and produces pathological lesions suitable for investigation of the pathophysiology of blunt head trauma.

Tumor necrosis factor-alpha augments radiation effects against human colon tumor xenografts.

Recent reports indicating that tumor necrosis factor-alpha (TNF-alpha) can augment the lethal effects of radiation against certain tumor cell lines prompted us to investigate whether this premise holds true for human colon tumor xenotransplants. Nude mice implanted s.c. with LS174T adenocarcinoma cells (day 0) were randomized into 4 groups: 1) no treatment; 2) TNF-alpha at 1 x 10(4) units/i.v. injection on days 1, 4, 8, and 10; 3) radiation at 4 Gy delivered on days 2, 5, 9, and 11; and 4) TNF-alpha + radiation administered using the same time-dose schedules as for groups 2 and 3. A decrease in tumor growth was obtained with radiation, but not TNF-alpha, as a single modality. However, significanty slower tumor growth was observed with TNF-alpha + radiation when compared to radiation alone. Blood and spleen cells from animals receiving both modalities exhibited the highest oxidative burst capacity. Histopathological evaluation showed large areas of necrosis in animals treated with radiation and with combined radiation + TNF-alpha, and only small areas of necrosis in animals treated with TNF-alpha alone. Necrosis in TNF-alpha-treated animals was not significantly larger than in controls. Irradiation of LS174T cells in culture generally decreased soluble TNF-alpha receptor and carcinoembryonic antigen in cell supernatants, but TNF-alpha was not detectable, regardless of radiation. The results show that pretreatment with TNF-alpha can significantly enhance the effects of radiation against human colon tumor xenografts and that the mechanisms of action may be related to increased oxygen radical production when both agents are administered and/or to induction of apoptosis by TNF-alpha. This data provides support for further investigations using TNF-alpha as an adjunctive agent in the radiotherapy of colon and other cancers.

Pilot study of monoclonal antibody localization in subcutaneous and intracranial lung tumor xenografts after proton irradiation.

The purpose of this study was to determine if proton irradiation can increase the localization of radiolabeled monoclonal antibodies (MAb) in subcutaneous (s.c.) or intracranial (i.c.) human lung tumors xenotransplanted in athymic rats. Rats with carcinoembryonic antigen (CEA)-expressing (NCI-H441) tumors were irradiated using 3 different proton time-dose regimens, followed by 111In-ZCE025, an anti-CEA MAb, which was injected 2 hr after the last dose of irradiation, and the animals were euthanized 3 days later for biodistribution and other assays. Proton irradiation at 10 gray (Gy) as a single dose or in 2 Gy fractions given on 5 consecutive days increased the uptake of 111In-ZCE025 into s.c. tumors by 292% and 182%, respectively, compared to nonirradiated controls. No enhancement in radiolabeled MAb delivery was seen after hemibrain irradiation in animals with i.c. tumors. Histopathological examination of both implantation sites showed a viable poorly differentiated adenocarcinoma with a decrease in blood vessel density, a decrease in mitotic activity, and an increase in areas of necrosis following irradiation as compared with adjacent nonirradiated tissue. CEA expression was generally maintained in vivo in that the marker was detectable in the tumor, plasma, and cerebrospinal fluid. Oxygen radical production by peripheral blood cells from s.c. and i.c. tumor-bearing rats exhibited strikingly different patterns of responsiveness. I.c. injected animals were 24% lighter than their s.c. injected counterparts, but no neurological signs of tumor progression were noted. The results indicate that proton irradiation can be used effectively to increase the delivery of radiolabeled MAb to s.c. implanted human lung tumor xenografts. However, in order to accomplish this in the brain, other radiation time-dose schedules and treatments may be needed.

Pericapillary arteriovenous malformations angiographically manifested as cerebral venous malformations.

Cerebral venous malformations have been diagnosed by angiographic features and are considered to be a benign anomaly. However, ample evidence indicates that stroke or similar symptomatology occurs in patients harboring a cerebral vascular malformation that was diagnosed angiographically as a venous malformation. The purpose of the study is to confirm the presence of a pericapillary arteriovenous malformation in these patients by analyzing the clinical history and surgical findings and correlating them with histological features. Thirteen patients were included in this study. Each patient fulfilled four criteria: 1. the patient was neurologically symptomatic; 2. the angiographic diagnosis was a venous malformation; 3. at operation, shunting arterioles (50-100 microns) were found to contribute to the malformation; and 4. histologically, a mixture of venous channels and arterioles with arterioles directly connected to venules was found. Based on the above findings, the malformation present in the 13 patients can be termed a 'pericapillary arteriovenous malformation'. Its angiographic distinction from the cerebral venous malformation requires technological advancement in the capability of magnifying images of arterioles and venules, along with improvement in image resolution.

Immunocytochemical binding to neurons of serum from spinocerebellar degeneration patients.

Antineural antibodies have been described in sera of patients with neurodegenerative disorders. We looked for the presence of those antibodies in the sera of patients with spinocerebellar degeneration. Serum IgG from four patients with familial spinocerebellar degeneration showed strong binding to cerebral cortical neurons, Purkinje cells, and dorsal root ganglia of normal human tissue sections stained with the peroxidase antiperoxidase (PAP) method at serum dilution of 1:500. No binding to neuroglia cells or cells of the granular layer of the cerebellum was seen. Sera from four immediate, asymptomatic relatives (son or sibling) showed only moderate binding to Purkinje cells and to dorsal root ganglia, but not to cortical neurons. Sera from seven patients with neurological diseases other than spinocerebellar degeneration and from five healthy subjects showed no binding to neural elements. The findings may be of value in the diagnosis and screening of patients suspected of having spinocerebellar degeneration; however, the significance of these antineural antibodies in the pathogenesis of spinocerebellar degeneration is uncertain and awaits further studies.

Mixed malignant mesenchymoma metastatic to the central nervous system.

A unique case of central nervous system metastases of mixed malignant mesenchymoma in an 84-year-old man is described. The tumor exhibited an osteogenic appearance in the cerebral lesion and a primitive mesenchymal appearance in lesions of the brain stem and cerebellum. The primary site was apparently the chest wall, and there were also metastases to the lung and liver.

Hydrocephalus: II. Cell number and size, and myelin content of the pre-shunted cerebral cortical mantle.

The number of cells in the brain and their size can be calculated by determining the total brain DNA, RNA and protein. Myelin can be determined biochemically. This analysis was applied to brains of a group of adult cats with craniectomies that were made severely hydrocephalilc by the introduction of kaolin or silicone into the basal cisterns. In severely hydrocephalic brains sectioned rostral to the area of kaolin inflammation there was no loss of dry weight. Wet weight increased slightly reflecting periventricular edema. There was a slight increase in total DNA, RNA and protein, presumably reflecting the increase in small inflammatory cells. There was a significant reduction in galactolipids reflecting myelin loss.

Needle aspiration cytology of intracranial lesions. A review of 84 cases.

Eighty-four needle aspirates of tumors and other intracranial lesions were diagnosed using smear preparations stained with Papanicolaou and Diff Quick stains. In 66 cases the cytologic diagnosis was compared with a histologic diagnosis on material removed during a craniotomy. In 92% of the cases the cytologic diagnosis agreed with the histologic diagnosis. In 21 cases the aspiration was performed under local anesthesia through a twist drill orifice. In none of these cases were there any complications secondary to the aspirations. The needle aspiration of intracranial lesions appears to be a safe and reliable diagnostic procedure.