Recovery of cellular activity in cryopreserved human foetal brain tissue.

Since long-term cryopreservation can cause losses in neural tissue viability and function a prerequisite would be the ability to monitor and promote functional recovery in donor tissue intended for neural transplantation. Rapid assessment of cryopreserved tissue's functional status prior to grafting is presently difficult in a clinical setting. A convenient indicator of functional status may be the level of DNA synthesis activity taking place in the tissue. Using immunocytochemical detection of incorporated bromodeox-yuridine we have quantified and compared DNA synthesis activity (expressed as proliferative capacity (PC)) in human foetal mesencephalic, striatal, cortical and cerebellar tissue before and after a 275-376 day storage in liquid nitrogen. There was a post-storage reduction in viability of 48-73% and in PC of 26-59%; the higher the PC before storage the greater the reduction after. Incubation of cryopreserved tissue with fetal calf serum resulted in 2-4-fold higher PC levels than serum-untreated controls and reached 80% of fresh tissue levels in mesencephalic cells after 3-4 h incubation. Assuming that quantification of proliferative activity is a practical indicator of the tissue's functional status, these findings suggest that treatment of the tissue with serum can largely restore the lost function caused by cryopreservation.

Enhanced in vitro survival and growth of foetal human mesencephalic dopaminergic neurones on laminin and collagen: implications for cell banking.

Culture of second trimester mesencephalic cells on laminin and collagen substrata has been investigated in an attempt to ascertain the effects of these extracellular matrix components on survival and growth of central dopaminergic (DA) neurones. There were 156.8-186.4% more cells attached to laminin and collagen than poly-D-lysine 6 h post-plating. By 24 h there was statistically no significant difference in the total number of cells attached to the three substrate but in terms of cell type-specific survival the proportion of mesencephalic DA neurones surviving on laminin and collagen substrata after 7 days in culture increased significantly compared with poly-D-lysine (1.4-1.6% versus 0.4% of the total cellular population), an effect augmented by bFGF treatment, which led to levels of 2% or more, with a concomitant decrease in the proportion of attritic DA neurones. These results indicate a critical requirement for ECM proteins in the survival and growth of in vitro-propagated central DA neurones at the time of plating and throughout the culture period. They also imply survival-enhancing interactions of ECM proteins and neurotrophic factors in developmental neuronal regulation and provide paradigms for obtaining high yields of these cells for neural transplantation cell banks.

Direct interaction with target-derived glia enhances survival but not differentiation of human fetal mesencephalic dopaminergic neurons.

Regulation of the developing nervous system involves attraction, guidance and modification of innervating neurons by target cells through diffusible and membrane-related factors. The trophic effects from specific cell types remain to be investigated and characterized. In a series of experiments in which human fetal mesencephalic dopaminergic cells were co-cultured with target or non-target neurons or glial cells in direct or contiguous contact, we demonstrate that striatal glial cells (target-derived glia) can enhance dopaminergic neuron survival by up to 400% compared to either non-target cell co-cultures or mesencephalic controls. When in direct contact with striatal neurons, a greater proportion of dopaminergic neurons had a more differentiated morphology. The enhancement of dopaminergic neuron survival by target-derived glia appears to be mediated both by direct contact, possibly through target membrane-specific phenomena, and by diffusible substances, whereas non-target glia appear to exert the trophic effects predominantly through the latter mechanism. The finding that target neurons influence mainly dopaminergic neuron differentiation and target glia their survival indicates multiple, target cell type-specific regulation of innervating neuron development. These findings also have relevance to the establishment of neuronal cultures for neural transplantation.

Susceptibility of human foetal brain tissue to cool- and freeze-storage.

Human second trimester foetal brain tissue was stored for a period of 1-6 weeks under various conditions in an attempt to evaluate factors influencing its susceptibility (cell loss) and survivability. Post-storage viability of mesencephalon, striatum, cerebellum and occipital cortex was assessed by a protocol combining vital staining with cell density counts so that tissue viability and cell loss could be evaluated simultaneously; tissue survivability was evaluated by cell culture. A significant amount of cell loss occurred after 24 h storage at room temperature, after one week at 4 degrees C and by two weeks at -20 degrees C in all structures; storage at -196 degrees C resulted in 17-21% cell loss at the end of a 6 week period. At -20 degrees C the cryoprotective effect of 20% FCS was equivalent to that of 15% FCS + 7% DMSO combined, suggesting potential use of serum in replacement of chemical additives. The procedure for removal of DMSO was critical to cell viability and survivability: single step dilution led to 27-39% greater cell loss than slow, multi-step dilutions. In comparison to fresh, non-stored tissue, immunocytochemical characterization of in vitro propagated stored tissue revealed no changes in the populations of major constituent cell types including neurones, dopaminergic neurones, glial and fibroblast cells. These results provide information on possible conditions under which transplant tissue can be satisfactorily stored depending on the prevailing requirements.

Brain implants in man do not break down the blood-brain barrier to dopamine and domperidone.

We have evaluated the blood-brain barrier (BBB) in 8 Parkinsonian patients before and after stereotactic implantation of foetal mesencephalon (STIM) and one patient with an adrenal medullary implant. Parenteral administration of dopamine did not reverse Parkinsonism pre-operatively or at 5 days, 1, 2, 3, 4 months and 1 year post-operatively. Apomorphine and domperidone reversed Parkinsonism and produced dyskinesia in all patients pre- and post-operatively. We conclude that the BBB remains intact to dopamine following implantation.

Stereotactic linac radiosurgery for arteriovenous malformations.

Stereotactic linear accelerator (linac) radiosurgery has been in operation in the West Midlands since 1987, the first of its kind in the United Kingdom. Forty two patients with high-flow cerebral arteriovenous malformations have been treated, 26 of whom have been followed up. Angiography one year after treatment showed that five lesions were obliterated, 11 were reduced in size and/or flow rate and 10 were unchanged. Overall results show that nine out of 10 patients reviewed at 24 months had total obliteration. Three patients had complications; one has fully recovered, one died of an unrelated cause at 36 months and the other died from recurrent haemorrhage at nine months. Two patients had recurrent non-fatal haemorrhage within 24 months of treatment; both recovered without further deficit. All patients are fit to work but eight are unemployed. Although the follow up period is short, the early results indicate a success rate similar to those published by others using linac radiosurgery.

Cerebral blood flow in patients with a subarachnoid haemorrhage during treatment with tranexamic acid.

Many clinicians currently use antifibrinolytic therapy (AFT) routinely in the management of subarachnoid haemorrhage (SAH). Many others do not, either because they remain unconvinced that AFT reduces the risk of rebleeding, or that the medication itself causes serious complications and in particular cerebral ischaemia. Nineteen randomly selected patients were studied, 9 receiving tranexamic acid (9 g a day) and the remaining 10 placebo, with SAH confirmed by CT scanning and by lumbar puncture. There was no difference between the active and placebo group regarding the age, sex, clinical grade, CT scan and angiographic appearance. The intravenous Xe133 technique was used for serial determinations of hemispheral cerebral blood flow. The cerebral blood flow remained stable during the first week following subarachnoid haemorrhage, and then fell progressively, reaching its bottom level by the end of the second week. The cerebral blood flow levelled out during the third week at the end of which a sharp elevation, well above the first week's post-subarachnoid haemorrhage level, was noted. This rebound rise of cerebral blood flow was observed for both cerebral hemispheres. Cerebral flow was greatest in the contralateral (to side of ruptured aneurysm) brain hemisphere save for the peak observed during the first week post-subarachnoid haemorrhage. The difference between the ipsi- and contralateral hemispheres was most pronounced in patients receiving active treatment. Analysis of variance showed that cerebral blood flow was reduced by the active treatment and especially more so on the ipsilateral side with the ruptured aneurysm. The usefulness of AFT should therefore be reconsidered.(ABSTRACT TRUNCATED AT 250 WORDS)

Visual evoked cortical responses and electroretinograms following implantation of human fetal mesencephalon to the right caudate nucleus in Parkinson's disease.

Pattern electroretinograms and visual evoked cortical responses following flash and checkerboard pattern reversal stimulation were performed in 7 patients with advanced Parkinson's disease before and after implantation of human fetal ventral mesencephalon to the head of the right caudate nucleus. Six to nine months following surgery there was bilateral enhancement of flash-evoked response amplitudes. Electroretinogram measurements were unchanged. Individual increases in amplitude and decreases in latency were seen following pattern stimulation which were not statistically significant. A relationship between change in dyskinesia, and change in checkerboard pattern stimulation amplitude at 2.4 cycles per degree suggests that amplitude effects at this stimulus frequency are mediated via altered dopaminergic receptor status in the grafted hemisphere. This may be relevant to the mechanisms leading to generalised clinical improvements following this grafting technique.

Pituitary adenoma proliferative indices and risk of recurrence.

Twenty unselected pituitary adenomas have been examined for proliferative indices (PIs), and anterior pituitary hormone expression. All but two of the tumours were non-functional with proliferative indices from less than 0.1 to 0.5%. Two tumours were null cell adenomas with PIs less than 0.1 and 0.2%. The PIs of the three recurrent tumours was less than 1.0%. Gonadotrophin and TSH immunoreactivity was heterogenous and was found in 12/20 (60%) of the tumours. There was no significant relationship between PI, hormone expression or any other measured parameter. The biological behaviour of pituitary adenomas with a PI of less than 0.1% is uncertain, but those with a PI of greater than 0.1% are more likely to recur although longer follow-up is needed to confirm this.

The effect of tranexamic acid on bleeding time and haemostasis.

The effect of tranexamic acid (TXA) on platelet function was examined in 37 patients with ruptured intracranial aneurysms. TXA was administered in a double-blind, placebo controlled manner to 37 patients with ruptured intracranial aneurysms. TXA was given to 17 patients in a dose of 9 g/24 hours, to another 6 patients in a dose of 6 g/24 hours; 14 patients were given a placebo. The effect of TXA on certain haemostatic function tests was evaluated between the treatment groups. We conclude that patients receiving TXA do not appear to be at risk of recurrent haemorrhage due to abnormal haemostasis.

Implantation of human fetal ventral mesencephalon to the right caudate nucleus in advanced Parkinson's disease.

Disaggregated ventral mesencephalic tissue from single aborted human fetuses of 11 to 18 weeks' gestation was implanted stereotaxically into a consistent striatal site in 12 patients with advanced Parkinson's disease. All were receiving optimum levodopa therapy and were examined preoperatively and at 3,6,9, and 12 months postoperatively. Immunosuppression was not used. There were significant sustained improvements at 12 months in three patients; motor fluctuations were absent in two. There were modest group improvements up to 6 months, with increased quality of "on" and "off" phases, quantity of on times, and specific improvements in contralateral upper limb bradykinesia. Preoperative levodopa requirements were reduced to a mean of 64% at 6 months and 61% at 12 months. Deterioration below baseline ratings occurred in three of nine patients who had consistent follow-up to 12 months. Grafting of midgestational human fetal tissue can lead to improvement in Parkinson's disease. Individual disease severity may be critical, and further trials are needed to identify host factors influencing outcome.

Alcohol consumption--a risk factor for hemorrhagic and non-hemorrhagic stroke.

PURPOSE: The risks of alcohol consumption and its association with stroke were studied in 621 patients with stroke and 573 control subjects using case-control methods. PATIENTS AND METHODS: Patients with stroke were subdivided into 193 with subarachnoid hemorrhage, 91 with intracerebral hemorrhage, and 337 with cerebral infarction. Data on recent alcohol consumption were obtained by questionnaire in patients with stroke and compared with data from an occupational screening survey in control subjects. RESULTS: Relative risks, adjusted for confounding variables, exhibited J-shaped associations with increasing levels of alcohol consumption classified into four categories--abstainer, 1 to 90 g, 100 to 390 g, and greater than or equal to 400 g weekly). The individual risks were 1, 0.7, 0.5, and 1.3 for subarachnoid hemorrhage; 1.0, 0.6, 0.5., and 2.5 for intracerebral hemorrhage, and 1.0, 0.6, 0.7, and 2.4 for cerebral infarction for men and women combined. CONCLUSIONS: The results suggest that low levels of alcohol consumption may have some protective effect upon the cerebral vasculature, whereas heavy consumption predisposes to both hemorrhagic and non-hemorrhagic stroke.

Seasonal variation of cerebrovascular diseases.

The seasonal variation in all admissions of all types of cerebrovascular disease within the West Midlands Region was examined between the years 1973-1980. There was a fluctuation for both sexes with a peak in winter, between the months of October and April; a trough was observed in late summer, in July and August. Multivariate analysis of the meteorological factors showed an association between hours of sunshine and intracerebral haemorrhage. The meterological variables were strongly correlated with each other making the selection of the most predictable variable to stroke difficult.

A comparative evaluation of clinical rating scales and quantitative measurements in assessment pre and post striatal implantation of human foetal mesencephalon in Parkinson's disease.

Six patients with advanced Parkinson's Disease were evaluated before and after implantation of human fetal ventral mesencephalic tissue to the head of the right caudate nucleus. The results of clinical assessment indicate that attempts to characterise patient fluctuations requires a combination of clinical rating scales and timing of specific limb tasks.

A series of experimental surgery for advanced Parkinson's disease by foetal mesencephalic transplantation.

12 patients with advanced Parkinson's Disease who had right caudate implantations of late stage foetal mesencephalon have been followed up for 1 to 2 years with extensive clinical and physiological assessments. Three patients failed to comply and were excluded. Seven of the remaining 9 patients showed substantial initial improvement which was well maintained in 4. Two of the 3 remaining patients of this group maintained lesser improvement. One returned to pre-operative state. Two patients with greatly advanced disease had only slight but brief improvement. A series of 24 matched patients have been treated and continue under investigation.

Fibrinolytic activity after subarachnoid haemorrhage and the effect of tranexamic acid.

Seventy-four patients with recent subarachnoid haemorrhage were randomly allocated to placebo or tranexamic acid treatment. Fibrinolytic activity in the blood and cerebrospinal fluid was assessed before treatment, one week later and two weeks later. The natural history of fibrinolysis following subarachnoid haemorrhage was obtained from analysis of the placebo group. Following subarachnoid haemorrhage, fibrin degradation products and plasminogen activity in the cerebrospinal fluid were elevated. Subsequently, fibrin degradation products in the cerebrospinal fluid fell progressively over the following 2 weeks. Changes in cerebrospinal fluid plasminogen activity correlated with those of blood plasminogen activity. Complications such as rebleeding, hydrocephalus or cerebral thrombosis could not be predicted from analysis of fibrinolytic activity. Tranexamic acid treatment resulted in a reduction in cerebrospinal fluid and blood plasminogen activity. The relevance of fibrinolysis in cerebrospinal fluid and blood to the management of subarachnoid haemorrhage is discussed.