✓ Intracarotid infusions of leukotriene C4 (LTC4) were used to selectively open the blood-tumor barrier in rats with RG-2 gliomas. Blood-brain and blood-tumor permeability was determined by quantitative autoradiography using 14C aminoisobutyric acid. Leukotriene C4 (4 µg total dose) infused into the carotid artery ipsilateral to the tumor increased twofold the unidirectional transfer constant for permeability within the tumor while no effect on permeability was seen in normal brain. No gamma glutamyl transpeptidase (γ-GTP) activity was seen in tumor capillaries in contrast to high γ-GTP in normal brain capillaries. These findings suggest that normal brain capillaries may resist the vasogenic effects of LTC4, while LTC4 will increase permeability in tumor capillaries. This could relate to the ability of γ-GTP to act as an enzymatic barrier and inactivate leukotrienes in normal brain capillaries. Intracarotid LTC4 infusion may be a useful tool to selectively open the blood-tumor barrier for delivery of antineoplastic compounds.
Keith L. Black, Wesley A. King and Kiyonobu Ikezaki
Lawrence S. Chin, Keith L. Black and Julian T. Hoff
✓ A patient with herniated thoracic discs in tandem is reported. The previous literature is reviewed. Difficulties with the preoperative diagnosis and the surgical approach to these lesions are discussed.
Joan L. Venes, Keith L. Black and Joseph T. Latack
✓ The authors report their recent experience with 14 meningomyelocele patients with the Arnold-Chiari II malformation. Three major types of fourth ventricle anomalies seen in the Arnold-Chiari II malformation are defined, based on preoperative magnetic resonance imaging and intraoperative ultrasound studies. The Type A deformity is defined as no cystic dilatation of the fourth ventricle. In the Type B anomaly, there is intracranial dilatation of the fourth ventricle. The Type C deformity involves intraspinal dilatation of the fourth ventricle, either dorsal to the cord or within the substance of the cord. The Type A deformity was most common in infants, and in two cases progression from a Type A to Type B deformity was documented. Recognition of the type of Arnold-Chiari II malformation aids in designing an operative approach more specific to that structural abnormality. Intraoperative ultrasound is a valuable adjunct in localization of the underlying anomalies and permits safe decompression of the fourth ventricle.
The authors' indications for surgery now include failure to thrive due to either early respiratory and swallowing dysfunction, progressive spasticity, or upper-extremity weakness. Nine patients significantly improved following surgery and three patients with a progressively deteriorating course were stabilized by surgery. Decompression of the fourth ventricle by fenestration and internal shunting appears to be well tolerated, even in young infants, and is recommended in the treatment of the Arnold-Chiari II deformity.
Keith L. Black, Kiyonobu Ikezaki and Arthur W. Toga
✓ Peripheral benzodiazepine receptor ligands were utilized to selectively image intracerebrally implanted C6 gliomas, RG-2 gliomas, and Walker 256 metastatic tumors by means of quantitative autoradiography. Intravenous injections of 3H-PK11195 (1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinoline carboxamide) or 3H-flunitrazepam in combination with clonazepam revealed high densities of peripheral benzodiazepine binding in glial tumors, with less binding in metastatic tumors. Peripheral binding was displaced by preadministration of excess PK11195. Topographical correlation was excellent between areas of histologically verified tumor and high densities of peripheral benzodiazepine binding. The choroid plexus, ependyma, and pineal gland also showed a moderate level of binding, but there was little binding in other normal brain structures or necrotic tissue. Binding densities were three- to fivefold higher in C6 glial tumors compared to normal cortex. Injection of 3H-flunitrazepam alone, which binds to both central and peripheral receptors, had the advantage of showing normal anatomic structures in addition to a clear definition of tumor topography. The potential value of peripheral benzodiazepine ligands in selectively imaging brain tumors in man with positron emission tomography is discussed.
David C. Waters, Julian T. Hoff and Keith L. Black
✓ The effect of standard parenteral nutritional formulas on cold-induced vasogenic edema formation in cats was examined and compared to the effects of 5% dextrose, 0.9% saline, and 40.5% mannitol. The amount of vasogenic edema formed during a 3-hour period of fluid infusion following cold injury was quantified by a computerized graphics tablet determination of the volume of Evans blue-dyed white matter. Specific gravity measurements were taken as a measure of white matter water content. Serum osmolality, urine output, arterial blood gases, hematocrit, body temperature, and systolic blood pressure were measured periodically throughout the infusion period. Parenteral nutritional formulas and a 40.5% mannitol solution produced greater changes in serum osmolality than did 5% dextrose or 0.9% saline. Greater changes in serum osmolality were associated with larger calculated volumes of edema in the injured hemisphere and lower water contents in the uninjured hemisphere. The data indicate that hyperosmolar solutions may potentiate vasogenic edema formation when the blood-brain barrier is open.
Takehiko Baba, Chung-Ching Chio and Keith L. Black
✓ To determine if leukotrienes are important mediators of vascular permeability in brain tumors, the effect of 5-lipoxygenase inhibitors on blood-tumor barrier permeability in rats harboring HK Walker 256 brain tumors was examined using quantitative autoradiography with α-14C-aminoisobutyric acid. The 5-lipoxygenase enzyme converts arachidonic acid to leukotrienes. Three 5-lipoxygenase inhibitors were utilized: BW755C, nordihydroguaiaretic acid, and AA-861. All three 5-lipoxygenase inhibitors significantly decreased vascular permeability both within the tumors and in brain adjacent to tumor. This suggests that capillary permeability in and adjacent to tumors is influenced by endogenous leukotrienes and that leukotrienes play an important role in brain tumor edema.
Chung-Ching Chio, Takehiko Baba and Keith L. Black
✓ The authors have previously reported that intracarotid infusion of 5 µg leukotriene C4 (LTC4) selectively increases blood-tumor barrier permeability in rat RG-2 tumors. In this study, rats harboring RG-2 tumors were given 15-minute intracarotid infusions of LTC4 at concentrations ranging from 0.5 µg to 50.0 µg (seven rats in each dose group). Blood-tumor and blood-brain barrier permeability were determined by quantitative autoradiography using 14C aminoisobutyric acid. The transfer constant for permeability (Ki) within the tumors was increased twofold by LTC4 doses of 2.5, 5.0, and 50.0 µg compared to vehicle alone (90.00 ±21.14, 92.68 ± 15.04, and 80.17 ± 16.15 vs. 39.37 ± 6.45 µl/gm/min, respectively; mean ± standard deviation; p < 0.01). No significant change in Ki within the tumors was observed at the 0.5-µg LTC4 dose. Blood-brain barrier permeability was selectively increased within the tumors. At no dose in this study did leukotrienes increase permeability within normal brain.
To determine the duration of increased opening of the blood-tumor barrier by LTC4 administration, Ki was measured at 15, 30, and 60 minutes after termination of a 15-minute LTC4 infusion (seven rats at each time point). The mean Ki value was still high at 15 minutes (92.68 ± 15.04 µl/gm/min), but declined at 30 minutes (56.58 ± 12.50 µl/gm/min) and 60 minutes (55.40 ± 8.10 µl/gm/min) after the end of LTC4 infusion.
Sulfidopeptide leukotrienes LTC4, LTD4, LTE4 and LTF4 were infused to compare their potency in opening the blood-tumor barrier. The mean leukotriene E4 was the most potent, increasing the permeability value 37½ fold compared with vehicle alone (139.86 ± 23.95 vs. 39.37 ± 6.45 µl/gm/min).
Keith L. Black, Takehiko Baba and William M. Pardridge
✓ Leukotriene C4 (LTC4) increases vascular permeability in systemic, brain tumor, and ischemic brain capillaries, but not in normal brain capillaries. This study examines whether the abundance of gamma-glutamyl transpeptidase (γ-GTP) in normal brain capillaries might act as an enzymatic barrier to vasoactive leukotrienes in the brain. Blood-brain barrier (BBB) permeability was determined by quantitative autoradiography using 14C-aminoisobutyric acid. Ischemia was produced by occluding the middle cerebral artery. Seventy-two hours after occlusion, γ-GTP activity in ischemic brain disappeared, and LTC4 (4-µg total dose), which was infused into the carotid artery ipsilateral to the occlusion, selectively increased permeability, Ki, approximately twofold within core ischemic tissue and adjacent tissue, compared to vehicle alone in seven brains (15.53 ± 6.03 vs. 7.29 ± 3.36, p < 0.05, and 8.76 ± 4.02 vs. 4.32 ± 2.65, p < 0.05, respectively). No effect on BBB was seen in nonischemic brain tissue. Twenty-four hours postocclusion, γ-GTP activity was still present, and LTC4 infusion did not increase permeability within ischemic tissue. However, inhibition of γ-GTP with acivicin allowed LTC4 to increase permeability even 24 hours after occlusion in ischemic core and adjacent tissue compared to vehicle alone in seven brains (17.21 ± 16.32 vs. 8.23 ± 6.58, p < 0.05, and 11.78 ± 7.96 vs. 4.56 ± 1.93, p < 0.01, respectively). Acivicin almost completely blocked both the histochemical activity of γ-GTP in brain capillaries and the metabolism of LTC4 in isolated bovine capillaries. These findings suggest that γ-GTP may help normal brain capillaries resist the vasoactive effects of LTC4. In contrast, γ-GTP is lost in injured brain capillaries, which allows LTC4 (in combination with other factors) to increase vascular permeability in ischemic brain and brain tumors.
Keith L. Black, K. Chen, Donald P. Becker and Jean E. Merrill
✓ In order to determine the in vivo immune response in glioblastoma, monoclonal and polyclonal antibodies specific for inflammatory leukocytes and immunoregulatory products were utilized to stain tissue from four surgical specimens. The more activated the inflammatory cells, the more activated the tumors appeared to be. In the tumor with the largest infiltration (Case 3), inflammatory cells were stained for interferon-γ interleukin-2. interleukin-1β. lymphotoxin, tumor necrosis factor-µ, and transforming growth factor-β. The tumor cells also expressed interleukin-1β, interleukin-6, transforming growth factor-β, tumor necrosis factor-µ, and prostaglandin E. In contrast, in the tumor with the least inflammatory response (Case 1), the tumor cells did not express any cytokines. Expression of cytokines by glioma cells was modest in the two cases with modest inflammatory responses. Cellular inflammation, primarily consisting of T cells and macrophages with few or no B cells or natural killer cells, was two- to 15-fold greater outside the tumor than within. In contrast to leukocytes outside the tumor, which were activated and expressing class II major histocompatibility antigens, leukocytes within the tumor parenchyma or at the tumor's edge were negative for these antigens. In the four specimens studied here, the tumor cells themselves were also negative for class II major histocompatibility antigens. These findings, although preliminary, suggest that inflammatory cells within gliomas are inactivated and that glioma cells may increase the expression of immunosuppressive cytokines in response to an increased lymphocyte infiltrate. This observation, if corroborated by more extensive studies, may help to explain the failure of immune treatments in glioblastoma multiforme.
Keith L. Black, Shan Hsu, Norman S. Radin and Julian T. Hoff
✓ Eicosapentaenoic acid (EPA) has been reported to improve postischemic cerebral blood flow (CBF). The present study was designed to determine whether sodium 5-(3′-pyridinylmethyl)benzofuran-2-carboxylate (U-63557A), a selective thromboxane synthetase inhibitor, could potentiate the effects of EPA on CBF in ischemic gerbils. Ischemia was produced by bilateral carotid artery occlusion for 15 minutes followed by reperfusion for 2 hours. Immediately after ischemia, gerbils were given either an intravenous bolus of 0.167 mg of EPA followed by a continuous infusion of EPA at 1 mg/hr, or U-63557A (10 mg/kg intraperitoneally), or U-63557A and EPA, or a saline infusion. Regional CBF was measured by the hydrogen clearance method, and brain water by the specific gravity technique. Brain prostaglandins were measured by radioimmunoassay. Preischemic CBF's ranged from 27.4 to 29.5 ml/100 gm/min for the four animal groups. After ischemia and 2 hours of reperfusion, CBF in the saline-infused gerbils was significantly decreased to 19.2 ml/100 gm/min. Gerbils treated with either EPA or U-63557A alone had a CBF of 23.7 and 21.6 ml/100 gm/min, respectively. Postischemic CBF in animals treated with both U-63557A and EPA was 30.0 ml/100 gm/min, significantly higher than in saline-infused gerbils. Brain levels of 6-keto prostaglandin (PG)F1α (the metabolite of PGI2) were significantly higher in gerbils treated with U-63557A and EPA compared to gerbils given EPA alone. This study indicates that U-63557A potentiates the effects of EPA on postischemic CBF. This is probably due to the ability of U-63557A to increase prostacyclin formation in the vessel wall.