In vitro neurogenesis by progenitor cells isolated from the adult human hippocampus.

Neurogenesis persists in the adult mammalian hippocampus. To identify and isolate neuronal progenitor cells of the adult human hippocampus, we transfected ventricular zone-free dissociates of surgically-excised dentate gyrus with DNA encoding humanized green fluorescent protein (hGFP), placed under the control of either the nestin enhancer (E/nestin) or the Talpha1 tubulin promoter (P/Talpha1), two regulatory regions that direct transcription in neural progenitor cells. The resultant P/Talpha1:hGFP+ and E/nestin:enhanced (E)GFP+ cells expressed betaIII-tubulin or microtubule-associated protein-2; many incorporated bromodeoxyuridine, indicating their genesis in vitro. Using fluorescence-activated cell sorting, the E/nestin:EGFP+ and P/Talpha1:hGFP+ cells were isolated to near purity, and matured antigenically and physiologically as neurons. Thus, the adult human hippocampus contains mitotically competent neuronal progenitors that can be selectively extracted. The isolation of these cells may provide a cellular substrate for re-populating the damaged or degenerated adult hippocampus.

Practical and technical aspects of trans-sphenoidal surgery.

Trans-sphenoidal surgery was first described more than a century ago. Today, this approach occupies a crucial place in the armamentarium of the neurosurgeon for the management of sellar, suprasellar, and parasellar pathological conditions. Over the years, the procedure has witnessed multiple modifications, benefitting from technological advances and from innovative ideas of pioneering neurosurgeons and otolaryngologists. The introduction of the microscope and then the endoscope allowed progressive improvement of visualization, illumination, and magnification in this restricted surgical corridor. With enhanced knowledge and understanding of the surgical anatomy of the skull base, the application of extended transsphenoidal approaches became possible, thus widening significantly the anatomical area that can be reached through this approach. In addition, the continuous improvement in imaging, image guidance, and microinstruments allowed better planning and precision during surgery. In sum, thanks to recent technological advance, trans-sphenoidal surgery can now be applied to a large area of the skull base and for a wide range of pathological conditions. With growing experience, the procedure is performed with enhanced safety and greater efficacy. In this paper, we review the historical evolution of trans-sphenoidal surgery and describe the modern applications and modifications of the procedure.

Staged use of the transsphenoidal approach to resect superior third ventricular craniopharyngiomas.

INTRODUCTION: Craniopharyngiomas are benign tumors, usually originating from the infundibulum or tuber cinereum. Their surgical treatment is challenging because of their relationship to neural and vascular structures. Large craniopharyngiomas that invade the upper third of the third ventricle are a common reason for patients to need a second operation to accomplish a gross total resection. Transsphenoidal approaches are being increasingly used in the treatment of craniopharyngiomas. Large craniopharyngiomas involving the superior third ventricle are most commonly resected through a staged approach, often involving a transcortical or interhemispheric route. CASE REPORT: The authors describe the use of an extended transsphenoidal approach as a second-stage operation to resect the intraventricular component of a large craniopharyngioma in an illustrative case. CONCLUSION: The authors find this to be an excellent indication for an endoscopic extended transsphenoidal approach in selected cases.

Spontaneous recanalization of the internal carotid artery resulting in thromboembolic occlusion of the ipsilateral ophthalmic artery and visual loss.

We report a 54-year-old man who suffered a stroke from a complete right internal carotid artery (ICA) occlusion. Two months later, he presented with right eye blindness. Imaging demonstrated 50% recanalization of his right ICA. He underwent a right carotid endarterectomy to prevent contralateral stroke from emboli through a patent anterior communicating artery. Recanalization of a completely occluded proximal ICA due to atherosclerotic disease has been reported but is rare, but such patients emphasize the importance of follow-up vascular studies.

Hypertrophic olivary degeneration after surgical removal of cavernous malformations of the brain stem: report of four cases and review of the literature.

BACKGROUND: Hypertrophic olivary degeneration (HOD) is a pathological phenomenon that occurs after injury to the dentato-olivary pathway. Its hallmarks include hypertrophy of the olive with increased T2 signal intensity on magnetic resonance imaging, and it often manifests with palatal tremor and oscillopsia clinically. METHOD: We report the cases of four patients who developed delayed HOD after surgical resection of pontine lesions. FINDINGS: We discuss the anatomical and pathological details of this disease and review the few other reported cases of HOD after resection of lesions within the brainstem. CONCLUSIONS: HOD should be recognized as a possible complication of surgery within the brainstem and must be diagnosed promptly so that patients can be appropriately counseled and symptoms can be treated.

Radiographic features of tumefactive giant cavernous angiomas.

BACKGROUND: Giant cavernous angiomas (GCAs) are very rare, and imaging features of GCAs can be very different from those of typical cavernous angiomas (CAs), making them a diagnostic challenge. The purpose of the study was to evaluate the radiographic features of GCAs, with an emphasis on the differentiating features from neoplastic lesions. METHODS: The neuroradiological findings of 18 patients who harbored a histologically verified GCA (CA of 4 cm or larger) were reviewed retrospectively. The magnetic resonance imaging (MRI) appearance, enhancement pattern, presence of edema or mass effect, size, and location of each lesion were recorded. When available, pertinent clinical information, including age, sex, and mode of presentation, was obtained. FINDINGS: Seizures, neurologic deficits, hemorrhage, and hydrocephalus were the most common presenting symptoms. The lesions were hyperdense and nonenhancing on computed tomography with frequent calcifications. On MRI, the lesions most commonly had a multicystic appearance, representing blood of various ages, and multiple complete hemosiderin rings. GCAs can present in any location with associating edema and mass effect, giving them a tumefactive appearance. No developmental venous anomaly was observed with any lesion. CONCLUSIONS: Most GCAs in our series presented as multicystic lesions with complete hemosiderin rings on MRI, giving a "bubbles of blood" appearance. Although this characteristic feature is helpful in the diagnosis of many cases of GCAs, the correct diagnosis in the remaining cases may not be apparent until histopathological evaluation of the specimen is made.

Incidental giant arachnoid granulation.

Arachnoid granulations may expand the dural sinuses or inner table of the skull. Although usually incidental, giant arachnoid granulations that are of sufficient size to fill the lumen of a dural sinus and cause local dilation or filling defects can rarely cause symptoms due to sinus obstruction leading to venous hypertension. This 31-year-old man presented with a 3-month history of progressive bifrontal headaches and a giant arachnoid granulation at the posterior superior sagittal sinus. Intrasinus pressure measurements showed no significant pressure difference across the lesion to explain the headaches, which were then treated medically. Dural sinus pressure measurement, in certain cases of giant arachnoid granulations, can be used to exclude the lesion as the cause of the patient's symptoms.

Remote cerebellar hemorrhage.

Remote cerebellar hemorrhage (RCH) is a rare but benign, self-limited complication of supratentorial craniotomies that, to the best of our knowledge, has not been described in the imaging literature. RCH can be an unexpected finding on routine postoperative imaging studies and should not be mistaken for more ominous causes of bleeding such as coagulopathy, hemorrhagic infarction, or cortical vein occlusion. Cerebellar hemorrhage in the typical setting can be identified as RCH and does not require more extensive or invasive evaluation.

Tamoxifen-induced enhancement of calcium signaling in glioma and MCF-7 breast cancer cells.

The antiestrogen tamoxifen is commonly used to treat breast cancer, but it also has therapeutic activity in several other types of cancer. Many of these tumors, including malignant gliomas, are estrogen receptor negative. Nonetheless, high concentrations of tamoxifen can directly reduce cell proliferation in some of these tumors and induce apoptosis. In this study, the role of tamoxifen in calcium signaling and calcium-induced cell death was studied in both malignant glioma cell lines and MCF-7 breast cancer cells. Tamoxifen potently increased the spatial expansion of calcium waves by 30-150% while significantly enhancing and prolonging agonist-induced calcium elevations. Furthermore, tamoxifen pretreatment accelerated calcium ionophore-induced death by more than 20 min, suggesting that tamoxifen lowered cellular resistance to calcium loads. In contrast to its potentiating of calcium signaling in tumors, tamoxifen had no significant effect on calcium signaling in cultures of primary astrocytes from either human or rat brain. This study demonstrates the existence of calcium signaling in breast cancer and glioma cells and identifies tamoxifen as a potential modulator of tumor-associated calcium signaling.

Direct gap junction communication between malignant glioma cells and astrocytes.

Gap junctions are intercellular channels that connect the interiors of coupled cells. We sought to determine the extent to which malignant glioma cells form gap junction channels with astrocytes from either adult human brain or rat forebrain. The astrocytic gap junction protein, connexin 43 (Cx43), was identified in immunoreactive plaques at areas of cell-to-cell contact between cocultured glioma cells and astrocytes. These gap junction plaques were composed of functional channels, because extensive dye coupling was evident between the glioma cells and astrocytes from both human and rat brain. Calcium signaling was also readily transmitted from glioma cells to astrocytes and vice versa. In live rat brain, injection of glioma cells prelabeled with the gap junction tracer, dicarboxy-dichlorofluorescein, revealed extensive dye transfer to host cells, demonstrating that malignant glioma cells directly couple with normal brain cells. These observations suggest that intercellular communication via gap junctions may play a role in regulating cellular interactions during tumor invasion. In fact, the presence of gap junctions between astrocytes and glioma cells was sufficient to induce a transformation of astrocytic phenotype. Astrocytes cocultured with C6 glioma cells overexpressing Cx43 were significantly smaller and expressed a lower level of glial fibrillary acidic protein than astrocytes cocultured with otherwise identical mock-transfected, gap junction-deficient C6 cells. Thus, direct cellular coupling with glioma cells result in a phenotypic transformation of astrocytes that may contribute to the susceptibility of surrounding tissue to glioma invasion.

Treatment of recurrent malignant gliomas with chronic oral high-dose tamoxifen.

The present clinical trial was undertaken to assess the clinical safety and possible efficacy of administering tamoxifen to patients with recurrent malignant glial tumors at dosages calculated to achieve levels sufficient to inhibit protein kinase C within the tumor cells. Chronic p.o. tamoxifen was administered in very high dosages to 32 patients (20 males and 12 females; age range, 26-75 years; mean, 49 years) with histologically verified malignant glioma [anaplastic astrocytoma (12 patients) or glioblastoma multiforme (20 patients)] who had demonstrated clinical and radiographical progression or recurrence following external beam radiation therapy (and additional chemotherapy in 11; immunotherapy in 2). The dosage of tamoxifen administered was 200 mg/day to males and 160 mg/day to females given in a twice daily schedule. Clinical and radiographical (defined as a greater than 50% decrease in volume of the enhancing lesion volume on magnetic resonance imaging and a decrease in metabolic activity on serial positron emission tomographic scans) response was noted in 8 patients (25%; 4/12 with anaplastic astrocytoma and 4/20 glioblastoma multiforme), with an additional 6 patients (19%) exhibiting stabilization of disease with minimal side effects. Median survival from the time of diagnosis for the entire cohort was 24 months (104 weeks), for the anaplastic astrocytoma group 42.5 months (185 weeks), and for the glioblastoma group 17.4 months (75.5 weeks). From the initiation of tamoxifen, median survival for the entire cohort was 10.1 months (44 weeks), for the anaplastic astrocytoma group 16 months (69 weeks), and for the glioblastoma group 7.2 months (31 weeks). The mean length of follow-up of all patients after initiating tamoxifen was 16 months (69 weeks), while the mean length of follow-up of alive patients is 22.6 months (98 weeks) (range up to 51 months). These data suggest that a subgroup of patients with malignant gliomas respond or stabilize with chronic high-dose tamoxifen therapy. This therapy may represent an alternative or adjuvant to existing chemotherapies for these tumors; further clinical trials are warranted.

Enhancement of radiosensitivity in human malignant glioma cells by hypericin in vitro.

Hypericin, an antidepressant and antiviral agent being evaluated in phase I and II trials for patients with HIV infection, is known to be a potent protein kinase C inhibitor. We have investigated its effects on cellular response to radiation via a tetrazolium-formazan cell growth rate assay using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and clonogenic assay in three human glioblastoma cell lines, U87-MG, A-172, and T98G, and a low-passage malignant glioma culture, 93-492. At a concentration of 5 microM, hypericin inhibited these cells slightly but caused significant radiosensitization (e.g., the cell survival rate after the radiation treatment was 50.2 and 26.0% in cells treated with 6 Gy and 6 Gy plus 5 microM hypericin in U87-MG cells, respectively; P = 0.0285). Hypericin also enhanced the radiosensitivity significantly in the low-passage glioma 93-492 cells. These findings suggest that hypericin represents a potential new agent in combination with radiation therapy of malignant gliomas.

Malignant glioma-derived soluble factors regulate proliferation of normal adult human astrocytes.

Malignant gliomas are characteristically surrounded by marked gliosis. To assess whether glioma-derived products contribute to the proliferation of astrocytes, a feature of the gliosis response, we evaluated the influence of culture supernatants from malignant human glioma lines and tumor cyst fluids collected from two patients with glioblastoma multiforme on the proliferation of non-transformed adult human astrocytes. Both the culture supernatants and cyst fluids significantly increased DNA synthesis in astrocytes as assessed by a double immunofluorescence glial fibrillary acidic protein-bromodeoxyuridine technique. The net proliferative effect mediated by glioma cell line supernatants was tumor growth phase-dependent, being preferentially expressed during the logarithmic phase of glioma cell growth. Specific growth factor molecules and cytokines known to be secreted by gliomas (epidermal growth factor, fibroblast growth factor, platelet-derived growth factor, transforming growth factor-beta, interleukin-6, and tumor necrosis factor-alpha) could not reproduce the mitogenic effects of the glioma-derived soluble factors. Cytokines which can induce DNA synthesis by adult human astrocytes in vitro, gamma-interferon and interleukin-1, were not detected in the culture supernatant of glioma lines used in this study. In conjunction with the documented effects of glioma products on endothelial and lymphoid cells, the current study suggests that soluble glioma products can contribute to the production of surrounding gliosis observed in vivo.

Protein kinase C inhibitors induce apoptosis in human malignant glioma cell lines.

Previous work has demonstrated the importance of the protein kinase C (PKC) system in regulating glioma growth, and has led to clinical trials utilizing PKC inhibitors as adjuncts in the therapy of patients harboring malignant gliomas. This study was performed to explore the possibility that inhibition of PKC in gliomas was triggering an apoptosis signal. Glioma cell lines were treated with PKC inhibitors staurosporine (10 nM), and tamoxifen (10 microM). DNA from cells treated with each of these drugs exhibited a 'ladder' pattern of oligonucleosome-sized fragments characteristic of apoptosis, thus suggesting that in glioma cells, these drugs may be cytocidal in action.

CSF neuron-specific enolase after methohexital activation during electrocorticography.

We report changes in CSF and serum neuron-specific enolase (NSE) before and after methohexital infusion during electrocorticography in three patients undergoing epilepsy surgery. NSE is a critical enzyme for energy metabolism that accounts for 1.5% of all soluble brain protein and is an accepted marker of neuronal injury. CSF NSE rose three- to fourfold from baseline within 60 minutes after methohexital activation. Serum NSE was unchanged. This report supports evidence that CSF NSE rises acutely after induction of epileptiform activity and suggests that CSF NSE is a marker of seizure activity.

Malignant glioma modulation of immune function: relative contribution of different soluble factors.

We analyzed a series of human glioma cell lines with regard to establishing what variables may contribute to their overall functional immunomodulating capability. We observed that supernatants derived from the gliomas, but not those from non-malignant human astrocyte cultures, suppressed lymphocyte proliferation. The extent of suppression elicited differed between tumors and for the same tumor depending upon its growth phase. For individual gliomas, supernatants from cultures approaching or at confluency elicited maximal lymphocyte suppression. For the series of tumors, levels of production of the immunosuppressive molecules transforming growth factor beta 2 and prostanoids (prostaglandin E2) did not correlate with the levels of functional suppression observed at any of the different growth phases. In some cases, glioma cultures grown in the presence of indomethacin to abolish prostanoid synthesis resulted in supernatants with net stimulatory activity. Our results indicate that malignant transformation of astrocytes is associated with acquisition of immunosuppressive capability which is determined by the combined effect of multiple immunomodulatory soluble factors, inhibitory or enhancing, and is dependent on the growth phase of the tumor.

Inhibition of human malignant glioma cell motility and invasion in vitro by hypericin, a potent protein kinase C inhibitor.

The effect of hypericin, an antiviral drug and a potent protein kinase C (PKC) inhibitor, on glioma cell invasion was investigated in vitro. Treatment of the established human glioblastoma cell line, T98G, with 1 microM hypericin for 24 h resulted in a significant inhibition of the cell invasion through an artificial basement membrane, but not cell attachment or proliferation. Furthermore, tamoxifen and staurosporine, both PKC inhibitors, also inhibited T98G cell invasion, suggesting that PKC may be the cellular target for hypericin-inhibited glioma cell migration. Similarly, hypericin decreased cell motility significantly in established lines, T98G and U87-MG, and also in a low-passage human malignant glioma cell line. Thus, hypericin may prove useful for studying mechanisms of glioma invasion, and may represent a new agent in malignant glioma therapy.

Growth inhibition and apoptosis in human neuroblastoma SK-N-SH cells induced by hypericin, a potent inhibitor of protein kinase C.

The effect of hypericin, an antiviral agent and inhibitor of protein kinase C (PKC), on cell proliferation and programmed death was investigated in the human neuroblastoma cell line SK-N-SH. Hypericin induced significant growth inhibition in a dose-dependent manner demonstrated by a microculture tetrazolium (MTT) assay. DNA isolated from cells treated with hypericin at concentrations over 1 microM exhibited a 'ladder' pattern of oligonucleosome-sized fragments characteristic of apoptosis. Similarly, treatment of the cells with the PKC inhibitors staurosporine, tamoxifen or phorbol ester PMA for 72 h also resulted in apoptosis, suggesting that hypericin may be triggering an apoptotic signal in neuroblastoma cells, which at least in part may be mediated by the inhibition of PKC.

Loss of Rb expression in an ACTH-secreting pituitary carcinoma.

Little is known about the molecular mechanisms of tumor progression in the pituitary. However, animal studies suggest that the Rb gene may be involved in the development of pituitary carcinoma. Pathologic examination of a pituitary tumor that included both benign and malignant components provided insight into this mechanism. Both benign and malignant tumors were immunoreactive for ACTH. The benign adenoma showed strong nuclear immunoreactivity for Rb, however, both the adjacent sellar carcinoma and its metastases were Rb-negative. This study suggests that loss of Rb may in some cases be important in the progression of pituitary adenoma to carcinoma.

Production of soluble autocrine inhibitory factors by human glioma cell lines.

Gliomas in vitro exhibit density-limited growth upon the attainment of confluency, an effect usually attributed to cell-cell contact inhibition. Since gliomas have been demonstrated to secrete an array of soluble factors which can enhance tumor growth, we undertook this study to ascertain whether production of soluble factors by the tumor may also inhibit growth in an autocrine manner, and whether production of such factors is associated with the growth phase of the glioma. We observed that cell-conditioned medium (supernatants) from non-confluent glioma cultures induced growth, while confluent culture supernatants produced pronounced growth suppression. These latter supernatants enhanced proliferation of non-transformed astrocytes. Supernatants derived from all stages of confluency produced inhibition of lymphocyte proliferation. To characterize these factors, dialyzed supernatant was tested and found to continue to produce lymphocyte suppression but no glioma growth limitation. Growth of tumors in indomethacin or in acetylsalicylic acid to abolish prostanoid synthesis abrogated the inhibitory influence on glioma growth but only partially reversed the lymphocyte suppressive capacity. These studies suggest that gliomas do produce a growth phase dependent autocrine inhibitory factor(s), and that the production of these small molecular weight factors is at least partially under control of the cyclooxygenase pathway.