✓ Nine human optic gliomas were examined in tissue culture. Typically, growth from the explants revealed well differentiated bipolar cells with abundant 9 to 10 nm fibers similar to those observed in the surgical specimens. Multinucleation was rare except for one culture, which had as many as 20 nuclei arranged in a palisading fashion along the periphery of some of the cells. Degenerative changes of the 9 to 10 nm fiber bundles with the production of amorphous electron-dense deposits were observed both in vivo and in vitro, and were thought to represent the formation of Rosenthal fibers. A distinctive feature of some of the optic gliomas was the ability of their long, thin cellular processes to form fibrous tangles in tissue culture. The correlation of these fibrous tangles in culture with Rosenthal fibers in vivo is still uncertain.
Robert L. Martuza, Paul L. Kornblith and Theodore M. Liszczak
Eben Alexander III, Peter McL. Black, Theodore M. Liszczak and Nicholas T. Zervas
✓ Irrigation of the subarachnoid space after aneurysmal subarachnoid hemorrhage (SAH) has been reported to alleviate subsequent arterial vasospasm. The authors have investigated the effect of lavage of the cerebrospinal fluid (CSF) space in the two-hemorrhage canine model of vasospasm. Twelve dogs had basilar cistern lavage with 120 cc of artificial CSF 24 hours after each of two SAH's, and 12 control dogs had two sequential SAH's without intervening lavage of clot. The amount of clot on the ventral brain stem was evaluated at sacrifice and was graded from 0 (no clot) to 4 (maximum clot) to assess the adequacy of clot removal. Dogs that had undergone lavage had a median grade of 1 (range Grade 0 to 2); control dogs had a median grade of 2 (range Grade 1 to 3.5, p < 0.001, Wilcoxon rank sum test), indicating significant reduction of gross clot by lavage. The neurological findings were graded from 0 to 5, based on meningismus, ataxia, paresis, and cranial nerve deficits. No significant differences in neurological grade were found on any day between the two groups.
Satisfactory angiograms were obtained before and 7 days after hemorrhage and were controlled for blood pressure and blood gases; these showed significant spasm in both groups. There was a mean reduction (± standard deviation) of 21.6% ± 16.2% in basilar artery diameter in control dogs, compared to a 28.8% ± 15.1% reduction in dogs with lavage (difference not significant, t-test). There was a strong, but insignificant, trend toward reduction of endothelial desquamation in the basilar and middle cerebral arteries in dogs with lavage compared to control animals (p = 0.06). Corrugation and tearing of the elastica, thickened intima, intimal fibroplasia, vacuolization of the endothelial or smooth-muscle cells, and presence of blood cells in the adventitia occurred similarly in both groups.
It appears that cisternal lavage 24 hours after hemorrhage in this model has no effect on the angiographic, neurological, or most morphological sequelae of SAH, in spite of evidence for removal of clot as seen at sacrifice. Any postulated interaction of clot and vessel resulting in chronic vasospasm must occur before this time. Evaluation of the effect of much earlier lavage (for instance, 1 hour after hemorrhage) may elucidate the point at which vasospasm is instigated after SAH, and help in determining what factors cause vasospasm.
Theodore M. Liszczak, Peter McL. Black, Argyris Tzouras, Lorraine Foley and Nicholas T. Zervas
✓ Experimental subarachnoid hemorrhage (SAH) was induced in adult rabbits by three repeated cisterna magna injections of autologous nonheparinized blood. One week after the last blood injection the animals were sacrificed and examined for morphological change.
No vasculopathy was noted in the basilar arteries and no periarterial thrombus was seen around the majority of these arteries. Gross and microscopic changes were observed in both the ventricles and choroid plexus. Ventricular changes included dilation of the lateral ventricles, destruction of ciliated ependymal cells, and deposition of small amounts of blood throughout the ventricular system. These ventricular changes are similar to the pathological sequelae of SAH in patients. Choroid plexus changes included electron-dense cytoplasmic inclusions and dilation of the lateral and subcellular spaces. The ventricular surface and the choroid plexus are both affected by the intracisternal injection of blood. The hydrocephalus that follows SAH may be potentiated by ependymal disruption and loss of ciliated activity in the ventricles. Changes in choroid plexus function may also occur.
Theodore M. Liszczak, Vassilios G. Varsos, Peter McL. Black, J. Philip Kistler and Nicholas T. Zervas
✓ Sequential cisternal blood injections in dogs reproduce some of the morphological and physiological features seen in man after subarachnoid hemorrhage-induced vasospasm. This study reports the morphological features observed in cerebral vessels in areas exposed to subarachnoid blood. Subarachnoid hemorrhage was produced in dogs by two cisternal injections of non-heparinized autologous blood 48 hours apart. Dogs were sacrificed 48 hours after the second injection. Angiographic narrowing of the basilar artery was routinely present 48 hours after the second injection, and there was a good correlation between angiographic vasospasm and a narrowed arterial lumen at postmortem examination. All basilar arteries showed structural changes with electron microscopic examination; these included endothelial cell vacuoles, early smooth-muscle cell necrosis, intimal changes, and adventitial erythrocytes, leukocytes, and mast cells. The finding that accompanied vessel constriction most uniformly was packing of the adventitial cerebrospinal fluid spaces with erythrocytes. Angiographically visible spasm was resistant to vasodilators. These data suggest that infiltration of blood elements into the arterial wall is an important concomitant feature of morphological and angiographic vasospasm.
Nicholas T. Zervas, Theodore M. Liszczak, Marc R. Mayberg and Peter McL. Black
✓ Cerebral blood vessels are devoid of vasa vasorum. Therefore, the authors have studied the microarchitecture of the adventitia of large feline cerebral vessels and systemic vessels of the same size, in an effort to determine how the vessels are nourished. The cerebral vessels contain a rete vasorum in the adventitia that is permeable to large proteins and is in continuity with the subarachnoid space. This substructure may be analogous to the systemic vasa vasorum and may contribute to the nutrition of the cerebral arteries.
Gerard M. Debrun, Vassilis Varsos, Theodore M. Liszczak, Kenneth R. Davis, Roberto S. Heros and Nicholas T. Zervas
✓ Experimental cervical carotid aneurysms in dogs are obliterated with isobutyl-cyanoacrylate (IBCA) injected under direct vision into the aneurysm. Reflux of IBCA into the artery was prevented by inflating either a latex or a Silastic balloon in the carotid artery at the level of the neck of the aneurysm. This balloon was introduced through a catheter advanced into the common carotid artery by femoral catheterization. The Silastic balloon was found to be much more effective than the latex balloon in preventing spillage of IBCA into the lumen.
Vassilios G. Varsos, Theodore M. Liszczak, Dae Hee Han, J. Philip Kistler, Juan Vielma, Peter McL. Black, Roberto C. Heros and Nicholas T. Zervas
✓ Angiographic spasm of cerebral arteries was produced in dogs by successive injections of cisternal blood 48 hours apart. Angiograms were taken before and after each cisternal injection. There was progressively greater angiographic vasospasm of the basilar artery. Intravenous aminophylline, 10 mg/kg/hr, reversed vessel constriction on the 1st and 3rd day after one injection of cisternal blood. On the 5th day after two blood injections (on Day 1 and Day 3), intravenous aminophylline, nifedipine (1 mg/kg), and intra-arterial bolus injection of 2 mg/kg papaverine failed to reverse the constriction. The intractable constriction produced in this model resembles that found in patients. The calcium antagonist, nifedipine, is as ineffective as the more traditional vasodilators in reversing vasospasm in this model.
Peter McL. Black, Paul L. Kornblith, Peter F. Davison, Theodore M. Liszczak, Linda P. Merk, Barry H. Smith, Paul E. McKeever and Eugene A. Quindlen
✓ This report presents the results of a study using multiple techniques of the established human cell line, LM, which has been developed in culture medium from a patient with a right temporoparietal glioblastoma. This cell line has human subtetraploid karyotype and has several features of a transformed line in culture. These include continuous propagation for 10 years, ability to form tumor nodules when transplanted into immunologically suppressed hamsters, and pleomorphic appearance. Ultrastructurally, it is characterized by multiple nuclei, few actin cables, and numerous surface-membrane microvilli, as well as abundant 9- to 10-nm cystoplasmic filaments. By its immunological reactivity, the line can be shown to contain glial fibrillary acidic protein at low levels, consistent with its glial origin and continued nature. Dibutyryl cyclic adenosine monophosphate (db-cAMP) induces formation of long astrocytic-like processes as well. Its membrane electrical characteristics include a low resting membrane potential and short time constant. Used in a microtiter antiglioma antibody cytotoxicity assay, LM yields a positive reaction to antibodies in the sera of 80% of patients with astrocytomas and only 9% of normal blood-bank donors, suggesting that it shares common antigens with other astrocytic tumor lines.
The varied characteristics of this glioblastoma-derived line emphasize the “multiforme” nature of the neoplasm and suggest that for characterization of any such line, multiple parameters are necessary to allow comparison with other long-term glioblastoma lines in the literature. The usefulness of the LM line in in vitro cell biological, immunological, chemotherapeutic, and radiobiological studies of gliomas makes such efforts very worthwhile.