The collection and processing of human brain tissue for research.

To further understand the neuroanatomy, neurochemistry and neuropathology of the normal and diseased human brain, it is essential to have access to human brain tissue where the biological and chemical nature of the tissue is optimally preserved. We have established a human brain bank where brain tissue is optimally processed and stored in order to provide a resource to facilitate neuroscience research of the human brain in health and disease. A donor programme has been established in consultation with the community to provide for the post-mortem donation of brain tissue to the brain bank. We are using this resource of human brain tissue to further investigate the basis of normal neuronal functioning in the human brain as well as the mechanisms of neuronal dysfunction and degeneration in neurodegenerative diseases. We have established a protocol for the preservation of post-mortem adult human brain tissue firstly by snap-freezing unfixed brain tissue and secondly by chemical fixation and then storage of this tissue at -80 degrees C in a human brain bank. Several research techniques such as receptor autoradiography, DNA and RNA analysis, are carried out on the unfixed tissue and immunohistochemical and histological analysis is carried out on the fixed human tissue. Comparison of tissue from normal control cases and from cases with neurodegenerative disorders is carried out in order to document the changes that occur in the brain in these disorders and to further investigate the underlying pathogenesis of these devastating neurological diseases.

Leptomeningeal infiltration as the presenting manifestation of a malignant glioma.

Infiltration of the leptomeninges by a malignant glioma typically occurs with recurrent supratentorial tumors, but patients may present with leptomeningeal gliomatosis before the primary tumor is diagnosed. This report describes two patients who presented with headache and signs of multifocal neurological disease. One of the patients had neurofibromatosis type I. In both patients the cerebrospinal fluid examination showed a mild pleocytosis, but malignant cells were not detected. The diagnosis of leptomeningeal gliomatosis was not confirmed until autopsy, but in retrospect imaging showed a small, asymptomatic primary tumor in both patients. Leptomeningeal gliomatosis should be considered in the differential diagnosis of chronic meningitis, if the patient is afebrile and if there are multifocal neurological signs, even when a primary tumor is not obvious.

Establishment of primary human meningiomas as subcutaneous xenografts in mice.

Meningiomas are the most frequently occurring benign central nervous system tumours. We determined whether a subcutaneous animal model of meningioma was feasible by implanting fresh meningioma tissue from six patients into 60 athymic (nude) mice, either as tissue blocks (38 mice) or as cell suspensions (22 mice). The tumour take-rates were 74% (block) and 50% (suspension), and the xenografts retained the original tumour grade and subtype morphology by light microscopy. Comparison of cell proliferation markers in xenografts and original tumours gave similar immunohistochemical score rates for Ki-67, but not for PCNA. With the exception of one atypical tumour surgical specimen, all tumours lacked p53 immunopositivity. Transmission electron microscopy of sections of tumour xenografts revealed ultrastructural features, including desmosomes and desmosome-like structures, characteristic of well-differentiated meningiomas. The xenografts grew progressively with a volume increase of more than 10-fold over 6-11 months and an apparent doubling time of 16 weeks. This study demonstrates the utility of the subcutaneous meningioma xenograft as a model for further biological and therapeutic studies.

Childhood cancer in New Zealand 1990 to 1993.

An epidemiological study of childhood cancer in New Zealand identified 409 children aged 0 to 14 years with malignant neoplasms newly diagnosed between 1990 and 1993 inclusive. The original microscopic material on which the diagnoses were based was reviewed in 398 cases and the neoplasms were allocated into the 12 major groupings and 48 further subcategories of the International Classification of Childhood Cancer (ICCC). The pathology reviewers agreed with group and subcategory classification of the confirmed cancers in all but one case of acute leukemia and three cancers of the central nervous system. Changes were also made in the FAB classification of three cases of acute non-lymphocytic leukemia and in the further subcategorisation of three Hodgkin's lymphomas and ten astrocytomas. The results show a high level of diagnostic accuracy for confirmed childhood neoplasms in that time period. Nine of 15 cases of malignant melanoma notified to the study were not confirmed for various reasons, which included a change in the pathological diagnosis in four cases. Compared with Victoria (Australia), New Zealand has a high incidence rate of lymphomas in boys and an unusual female preponderance of Wilms' tumor cases.

Low-grade supratentorial astrocytomas in adults: Management, immunohistochemical analysis and long-term follow-up.

A retrospective review of supratentorial, low-grade astrocytomas in adults over a 10-year period (1983-1993) was performed. All 62 patients had computed tomography and surgery with histological and immunohistochemical analysis of the tumour. Radiotherapy was administered to all patients, regardless of the extent of surgical resection. Multivariate analysis showed that age of the patient at the time of surgery (P=0.008) and female sex (P=0.031) were the most important indicators of improved survival. No significant survival advantage was found with any particular symptom or symptom duration. Neither presence of a tumour cyst nor site of the tumour affected survival. Histological grading (St Anne-Mayo system) found 61 grade II astrocytomas (98%) and one grade I tumour. Six tumours (10%) were protoplasmic astrocytomas and 56 cases (90%) were diffuse fibrillary astrocytomas. Proliferation index derived from Ki-67 antibody staining and the presence of p53 protein expression were not significantly correlated with survival. Most patients underwent biopsy (79%) with 8% undergoing subtotal and 13% total resection. Increased extent of surgical removal showed a trend towards an improved survival (P=0.05). No survival advantage was found with increasing radiotherapy dose. Median survival in the study population was 5.1 years, with survival rates of 70% at 2 years and 53% at 5 years. The follow-up period ranged from 1 to 10.5 years. Younger age, female sex and the extent of surgical resection are important prognostic factors in the management of low-grade astrocytomas, whereas the efficacy of postoperative radiotherapy needs further evaluation. The prognostic significance of Ki-67 and p53 expression in low-grade astrocytomas remains to be determined.

Brain-derived neurotrophic factor is reduced in Alzheimer's disease.

Alzheimer's disease may be due to a deficiency in neurotrophin protein or receptor expression. Consistent with this hypothesis, a reduction in BDNF mRNA expression has been observed in human post-mortem Alzheimer's disease hippocampi. To further investigate this observation, we examined whether the alteration in BDNF expression also occurred at the protein level in human post-mortem Alzheimer's disease hippocampi and temporal cortices using immunohistochemical techniques. We observed a reduction in the intensity and number of BDNF-immunoreactive cell bodies within both the Alzheimer's disease hippocampus and temporal cortex when compared to normal tissue. These results support and extend previous findings that BDNF mRNA is reduced in the human Alzheimer's disease hippocampus and temporal cortex, and suggest that a loss of BDNF may contribute to the progressive atrophy of neurons in Alzheimer's disease.

Insulin-like growth factor-I (IGF-I) immunoreactivity in the Alzheimer's disease temporal cortex and hippocampus.

IGF-I has been shown to enhance neuronal survival and inhibit apoptosis. IGF-I immunoreactivity was examined in the Alzheimer's disease and normal post-mortem human hippocampus and temporal cortex to determine whether IGF-I protein levels are altered in response to neurodegeneration. IGF-I immunoreactivity was induced in a subpopulation of GFAP-immunopositive astroglia in the Alzheimer's disease temporal cortex. These observations raise the possibility that IGF-I has a neuroprotective role in the Alzheimer's disease brain.

Expression of Fos, Jun, and Krox family proteins in Alzheimer's disease.

Apoptosis is an active process of cell death characterized by distinct morphological features and is often the end result of a genetic program of events, i.e., programmed cell death (PCD). There is growing evidence supporting a role for apoptosis and/or PCD in Alzheimer's disease (AD), based on DNA fragmentation studies and recent findings of increased levels of inducible transcription factors (ITFs) such as c-Jun in AD brains. We have characterized the expression of a large range of ITFs (c-Fos, Fos B, Fos-related antigens, c-Jun, Jun B, Jun D, Krox20, and Krox24) using multiple antisera in AD postmortem hippocampi and compared this with human control hippocampi as well as Huntington's disease hippocampi and human epilepsy biopsy tissue. We found little evidence of nuclear expression of any ITF except c-Jun in the human postmortem tissue, compared with nuclear staining in biopsy tissue. We found some evidence for increased levels of c-Jun and Krox24 protein and krox24 mRNA in the CA1 region of AD hippocampi, suggesting that PCD may be involved in the pathogenesis of AD. In general, staining characteristics of ITFs varied with different antisera directed against the same protein, indicating the need for caution when interpreting results.

Loss of A1 adenosine receptors in human temporal lobe epilepsy.

Using quantitative receptor autoradiographic methods we have examined A1 adenosine receptors, adenosine uptake sites, benzodiazepine receptors, NMDA, AMPA, and kainic acid receptors in temporal lobes removed from patients suffering from complex partial seizures and in normal control post-mortem temporal cortex. Binding to A1 adenosine receptors and NMDA receptors was reduced in epileptic temporal cortex, while the other neurochemical parameters were unchanged. The reason for this A1 receptor loss is unclear as it occurred in both idiopathic and symptomatic cases and thus may be a consequence rather than an initial cause of seizures. However, because adenosine is a powerful anticonvulsant substance, loss of anticonvulsant A1 receptors may contribute to the human epileptic condition. It is also possible that the observed differences in A1 binding are due to autopsy vs. biopsy changes in the levels of A1 adenosine receptors.

Leptomeningeal and brain biopsy in chronic meningitis.

BACKGROUND: The cause is not found in one third of patients presenting with chronic meningitis. Biopsy of the leptomeninges and brain is often recommended in these patients, but the value of a biopsy is uncertain. AIMS: To review the results of leptomeningeal and brain biopsies and their effect on diagnosis and management in patients with chronic meningitis, if the diagnosis was uncertain after clinical assessment, lumbar puncture and radiological investigations. METHODS: The clinical features, investigations and the results of leptomeningeal and brain biopsies were reviewed retrospectively in 25 patients presenting with chronic meningitis between 1967 and 1990. RESULTS: The biopsy identified the cause of the chronic meningitis in five patients (neoplastic meningitis in three, tuberculous meningitis in one, granulomatous angiitis in one). In 17 patients the biopsy was abnormal but it did not identify the cause (non-granulomatous lymphocytic meningitis in eight, granulomatous meningitis in two, non-specific abnormalities in seven). In two patients the biopsy was normal and in one patient the tissue was used only for culture. The results of the biopsy led to a beneficial change in treatment in two patients but did not influence management in the other patients. CONCLUSIONS: A leptomeningeal and brain biopsy was of limited practical value in diagnosis and management in most patients with chronic meningitis.

A fatal case of ovarian hyperstimulation syndrome with cerebral infarction.

The ovarian hyperstimulation syndrome (OHSS) as a cause of death in infertile patients involved in in vitro fertilization is an extremely rare phenomenon. Reported here are the clinical and pathological feature of just such a case together with a discussion of the pathophysiology thought to be involved.

Diagnostic markers demonstrating successful culture of human meningioma cells.

The light microscopic and ultrastructural features of cultured meningioma cells were compared with those of surgically resected specimens. Meningioma cells normally show variable degrees of epithelial or mesenchymal differentiation. In cultured cells, well formed psammoma bodies and whorls were seen by light microscopy and specialised intercellular junctions were observed by electron microscopy. Cytogenetic analysis was also shown to be a useful diagnostic marker to differentiate cultured meningioma from contaminant proliferating fibroblastic and endothelial cells. These methods have demonstrated that the cells cultured from operative samples were predominantly meningioma cells. The ability to reproducibly cultivate populations of meningioma cells should facilitate in vitro assessment of potential adjunctive treatment modalities.

Chronic lithium neurotoxicity presenting as Parkinson's disease.

A 71 year old man who had been on lithium for 9 years for mania presented with an encephalopathic illness which was almost certainly due to lithium intoxication. Having recovered from this acute episode (although he was left with some sequelae) he was recommenced on lithium for his manic symptoms with a careful control of his blood levels. After remaining fairly stable for 8 years he presented with features suggestive of Parkinsonism and was admitted to hospital for investigation. There was no history of taking additional medication such as antidepressants or antipsychotics. He died in hospital and a post-mortem examination confirmed the cause of death as acute myocardial infarction. However histological examination of the brain revealed neurological sequelae of chronic lithium intoxication. There was no evidence of degenerative condition such as Parkinsonism or Alzheimer's disease.

Brain biopsy in the management of focal encephalitis.

The value of a brain biopsy in diagnosis and management of suspected herpes simplex encephalitis was studied in 29 patients (16 prospectively and 13 retrospectively). The biopsy showed herpes simplex encephalitis in eight, culture-negative encephalitis in 14, and was normal in three patients. It provided an alternative diagnosis in four patients, for two of whom curative treatment was available. The biopsy was complicated by a fatal intracranial haemorrhage in one patient. The low yield of alternative diagnoses suggests that a brain biopsy is not justified in the routine investigation of focal encephalitis.

The acromegalic rosary.

Palpable enlargement of the costochondral junctions on the anterolateral chest wall was detected in 26 of 27 patients with acromegaly. In a further study three doctors wearing eyemasks examined a further 13 acromegalic patients and 12 control subjects; costochondral enlargement was detected with a sensitivity of 77% and specificity of 86%. It was not found in normal subjects in the absence of previous chest injury. The presence and extent of costochondral enlargement was unrelated to the duration of active or cured acromegaly, the initial or present growth hormone concentrations, or other clinical features of growth hormone excess. Costochondral enlargement is a common clinical sign in acromegaly and can serve as a biological marker of previous or current growth hormone excess.

Delayed seizures occurring with hypoxic-ischemic encephalopathy in the fetal sheep.

The pathogenesis of perinatal hypoxic-ischemic encephalopathy is poorly understood. Most insults are thought to occur before or during birth. We have investigated the evolution of parasagittal EEG activity and distribution of neuronal damage after cerebral ischemia in chronically instrumented fetal sheep (119-126 d gestation). The vertebral-carotid anastomoses were ligated and cerebral ischemia was induced by inflating occluder cuffs around the carotid arteries for 30 min. Parietal cortical EEG activity was analyzed with real-time spectral analysis with reference to control fetuses. After ischemia, EEG activity was suppressed, then rapidly increased in intensity at 8 +/- 1 h to a peak at 9 +/- 1 h postischemia. There was increased intensity of the lower frequencies (1-7 Hz) apparent as epileptiform activity with convulsions. This low-frequency hyperactivity gradually resolved by 28 +/- 7 h postinsult. After 72 h, the loss of intensity at all frequencies and laminar necrosis of the underlying parasagittal cortex indicated irreversible brain injury. Ranking the structures in order of decreasing amounts of damage: parasagittal cortex greater than hippocampal CA1, 2, and 3 regions greater than lateral cortex, hippocampal CA4 region and striatum greater than amygdala, dentate gyrus, thalamus, and cerebellum. The evolution of EEG activity and the distribution of damage after cerebral ischemia closely resembles the time course and pathology of hypoxic-ischemic encephalopathy seen in some severely asphyxiated term neonates. The consistent electrophysiologic and histologic outcome should allow this experimental approach to be valuable in testing a number of current hypotheses relating to perinatal asphyxial encephalopathy.

Alzheimer's disease: changes in hippocampal N-methyl-D-aspartate, quisqualate, neurotensin, adenosine, benzodiazepine, serotonin and opioid receptors--an autoradiographic study.

The following receptors were assessed post-mortem in the hippocampi (anterior region) of eight patients with Alzheimer's disease and nine age-matched controls, using autoradiography: N-methyl-D-aspartate (including glutamate, phencyclidine and glycine binding sites), quisqualate, kainic acid, adenosine A1, benzodiazepine, serotonin (1 and 2), muscarinic cholinergic, beta-adrenergic, neurotensin and opioid receptors. In CA1 there were significant parallel losses of binding to the three N-methyl-D-aspartate-linked sites (average reduction 46%) and also losses of quisqualate (38%) and serotonin2 (58%) receptor binding, with a 47% loss of binding to A1 sites. Binding to all of these receptors was also reduced in CA3 (except binding to A1 sites which was normal) but only the serotonin2 receptor binding loss reached significance (52%). A significant reduction in binding was also observed in the entorhinal area to the N-methyl-D-aspartate receptor-linked sites (average reduction = 39%), benzodiazepine (40%) and serotonin2 receptors (45%), and there was a loss of binding to neurotensin (57%) and opioid receptors (42%). Significant reductions in the dentate gyrus molecular layer were seen for serotonin2 receptors (44%), and binding to opioid (44%) and A1 receptors (46%). Levels of ligand binding to muscarinic cholinergic, serotonin1, beta-adrenergic and kainic acid receptors were not significantly different from control values in any of the four areas examined. These results provide support for observations of selective receptor changes in Alzheimer's disease involving a broad range of receptor types which encompass both excitatory amino acid and other receptors (notably serotonin2, A1, benzodiazepine, neurotensin and opioid receptors). The implications of the pattern of receptor changes for the suggestion that excitotoxicity plays a role in the disease are discussed, as is the possible contribution of the receptor changes to the symptomatology of Alzheimer's disease.

Adrenoleukodystrophy: manifestations of a protean disorder: report of a recently identified kindred.

Adrenoleukodystrophy is an inherited metabolic disorder resulting in accumulation of fatty acids in body tissues, principally the central nervous system. We report a recently identified kindred with this disease to highlight the varied clinical manifestations and recent advances in diagnosis and treatment.