Analysis of loss of heterozygosity on chromosome 10 in patients with malignant astrocytic tumors: correlation with patient age and survival.

OBJECT: The most frequent genetic abnormality in human malignant gliomas is loss of heterozygosity (LOH) on chromosome 10. Candidate genes on chromosome 10 that are associated with the prognosis of patients with anaplastic astrocytoma (AA) and glioblastoma (GBM) were evaluated. METHODS: The authors used 12 fluorescent microsatellite markers on both arms of chromosome 10 to study LOH in 108 primary astrocytic tumors. The LOH on chromosome 10 was observed in 11 (32%) of 34 AAs and 34 (56%) of 61 GBMs. No LOH was detected in 13 low-grade gliomas. Loss of heterozygosity was not detected in any AA in the seven patients younger than 35 years, but it was discovered in 41% of the patients older than 35 years. The prognostic significance of LOH at each locus was evaluated in 89 patients older than 15 years; 33 (37%) had supratentorial AAs and 56 (63%) had supratentorial GBMs. The Cox proportional hazards model, adjusted for patient age at surgery, the preoperative Karnofsky Performance Scale score, and the extent of surgical resection revealed that LOH on marker D10S209 near the FGFR2 and DMBT1 genes was significantly associated with shorter survival in patients with AA. The LOH on markers D10S215 and D10S541, which contain the PTEN/MMAC1 gene between them, was significantly associated with shorter survival in patients with GBM. CONCLUSIONS: In the present study it is found that LOH on chromosome 10 is an age-dependent event for patients with AAs and that LOH on marker D10S209 near the FGFR2 and DMBT1 loci is a significantly unfavorable prognostic factor. It is also reported that LOH on the PTEN/MMAC1 gene is a significantly unfavorable prognostic factor in patients with GBM.

Radiosurgery-induced microvascular alterations precede necrosis of the brain neuropil.

OBJECTIVE: Radiosurgery is used as a therapeutic modality for a wide range of cerebral disorders. It is important to understand the underlying causes of deleterious side effects that may accompany gamma-irradiation of brain tissue. In this study, structural alterations in rat cerebral vessels subjected to gamma knife irradiation in vivo were examined, for elucidation of their potential role in necrosis formation. METHODS: A maximal center dose of 75 Gy was delivered to the rat parietal cortex with a 4-mm collimator, and changes occurring before necrosis formation were assessed 3.5 months after irradiation. Transmission electron microscopy, using horseradish peroxidase as a tracer, and scanning electron microscopy with vascular casting were performed. RESULTS: The capillary network in the irradiated area exhibited thickening and vacuolation of the basement membrane. The capillary density in the irradiated area was lower and the average capillary diameter was larger, compared with the nonirradiated side. These results indicate that substantial changes in the neuropil do not occur 2 weeks before the time of definite necrosis formation, whereas changes in the basement membrane are prominent. CONCLUSION: The necrotic response to intermediate doses of focused-beam irradiation appears after a considerable latency period and then progresses rapidly. This contrasts with previously reported responses to fractionated whole-brain irradiation, in which damage occurs slowly and gradually. Alterations in the microvascular basement membrane precede overt cellular changes in neuronal and vascular cells and provide an early index of cerebrovascular dysfunction in regions destined to undergo necrosis.

De novo aneurysm formation after stereotactic radiosurgery of a residual arteriovenous malformation: case report.

We report a case of a 19-year-old woman who underwent radiosurgical treatment of a residual arteriovenous malformation. Nine months after treatment, repeat angiography revealed a de novo paranidal aneurysm that was treated endovascularly. We postulate that changes in flow dynamics or vessel integrity after radiosurgery contributed to the formation of her de novo aneurysm.

A stereotactic device for experimental rat and mouse irradiation using gamma knife model B--technical note.

BACKGROUND: For radiobiological experiments using the Gamma Knife model B, we constructed a stereotactic device to irradiate rat and mouse brains and verify the absorbed dose at the target using thermoluminescence dosimetry and a head phantom. METHODS: Our stereotactic device is primarily designed for rats using the fixation principles of a stereotactic atlas. A head-fixation adapter for a mouse was constructed to enable targeted irradiation of mouse brains. We built simple phantoms to simulate rat and mouse heads. We placed thermoluminescent dosimeters at various positions on the phantom for dose measurements. Dose planning employed the Leksell Gamma Plan version 4.11 software, assuming a spherical skull geometry for all calculations. FINDINGS: The measurements demonstrated that the actual absorbed dose agreed with our calculations within the errors of thermoluminescence dosimetry and the accuracy of our irradiation technique and dose calculations. INTERPRETATION: This device provides an accurate method for irradiating rat and mouse brains using the Gamma Knife model B.

Anticonvulsant effects of gamma surgery in a model of chronic spontaneous limbic epilepsy in rats.

OBJECT: The management of intractable epilepsy remains a challenge, despite advances in its surgical and nonsurgical treatment. The identification of low-risk, low-cost therapeutic strategies that lead to improved outcome is therefore an important ongoing goal of basic and clinical research. Single-dose focal ionizing beam radiation delivered at necrosis-inducing and subnecrotic levels was investigated for its effects on seizure activity by using an established model of chronic recurrent spontaneous limbic seizures in rats. METHODS: A single 90-minute period of repetitive electrical stimulation (inducing stimulus) of the hippocampus in rats elicited a single episode of status epilepticus, followed by a 2- to 4-week seizure-free period. Spontaneous recurrent seizures developed subsequently and persisted for the duration of monitoring (2-10 months). Simultaneous computerized electroencephalography and video recording were used to monitor the animals. After the establishment of spontaneous recurrent seizures, bilateral radiation centered in the ventral hippocampal formation was administered with the Leksell gamma knife, aided by a stereotactic device custom made for small animals. A center dose of 10, 20, or 40 Gy was administered using a 4-mm collimator. Control animals were subjected to the same seizure-inducing stimulus but underwent a sham treatment instead of gamma irradiation. In a second experiment, the authors examined the effects of gamma irradiation on the proclivity of hippocampal neurons to display epileptiform discharges. Naive animals were irradiated with a single 40-Gy dose, as already described. Slices of the hippocampus were prepared from animals killed between 1 and 178 days postirradiation. Sensitivity to penicillin-induced epileptiform spiking was examined in vitro in slices prepared from control and irradiated rat brains. CONCLUSIONS: In the first experiment, single doses of 20 or 40 Gy (but not 10 Gy) reduced substantially, and in some cases eliminated, behaviorally and electrographically recognized seizures. Significant reductions in both the frequency and duration of spontaneous seizures were observed during a follow-up period of up to 10 months postradiation. Histological examination of the targeted region did not reveal signs of necrosis. These findings indicate that single-dose focal ionizing beam irradiation at subnecrotic dosages reduces or eliminates repetitive spontaneous seizures in a rat model of temporal lobe epilepsy. In the second experiment, synaptically driven neuronal firing was shown to be intact in hippocampal neurons subjected to 40-Gy doses. However, the susceptibility to penicillin-induced epileptiform activity was reduced in the brain slices of animals receiving 40-Gy doses, compared with those from control rats that were not irradiated. The results provide rational support for the utility of subnecrotic gamma irradiation as a therapeutic strategy for treating epilepsy. These findings also provide evidence that a functional increase in the seizure threshold of hippocampal neurons contributes to the anticonvulsant influence of subnecrotic gamma irradiation.

A methodology designed to increase accuracy and safety in stereotactic brain surgery.

A series of technical tips and devices designed to increase accuracy and safety in stereotactic surgery are presented. We use stereotactic magnetic resonance imaging with three-dimensional magnetization-prepared rapid gradient-echo (MP-RAGE) imaging to minimize image distortion, and a three-dimensional stereotactic planning system for accurately registering three-dimensional space. We also developed several technical devices useful for stereotactic intracranial procedures; an applicator system attached to the frame which simulates the fiducial markers in order to keep the target at a suitable position in stereotactic space; a torque wrench to set the torque on the fixing pins to the frame reproducibly at 5 inch pounds in order to keep distortion of the frame to a minimum while maintaining secure fixation; an entry point marker to maintain the calculated trajectory angle; a straightening cannula to prevent the thermo-coagulation needle from bending; a microvascular Doppler and its holder to detect significant vessels and to know their precise depth in order to avoid vascular injury from thermocoagulation; a burr hole button device to secure depth electrode cables at the patient's skull.

Familial arteriovenous malformations in siblings.

BACKGROUND: Familial arteriovenous malformations (AVMs) of the brain are rare. We present two sisters with the same parents who harbored AVMs that were successfully treated. METHODS: The elder sister presented with a unilateral migrainous type of headache overlying the right parietal area. The younger one suffered from exercise-induced headaches. Both were neurologically intact. Magnetic resonance imaging scans of the brain and cerebral angiography delineated the lesions. Both sisters underwent endovascular embolization followed by surgical resection. RESULTS: Postoperatively, aside from a left inferior quadrantanopsia in the elder sister, both were neurologically intact. CONCLUSIONS: We report the rare occurrence of familial AVMs in two siblings and review the literature of 14 reports. No genetic predisposition was found.

Immunological detection of alkaline-diaminobenzidine-negativeperoxisomes of the nematode Caenorhabditis elegans purification and unique pH optima of peroxisomal catalase.

We purified catalase-2 of the nematode Caenorhabditis elegans and identified peroxisomes in this organism. The peroxisomes of C. elegans were not detectable by cytochemical staining using 3, 3'-diaminobenzidine, a commonly used method depending on the peroxidase activity of peroxisomal catalase at pH 9 in which genuine peroxidases are inactive. The cDNA sequences of C. elegans predict two catalases very similar to each other throughout the molecule, except for the short C-terminal sequence; catalase-2 (500 residues long) carries a peroxisomal targeting signal 1-like sequence (Ser-His-Ile), whereas catalase-1 does not. The catalase purified to near homogeneity from the homogenate of C. elegans cells consisted of a subunit of 57 kDa and was specifically recognized by anti-(catalase-2) serum but not by anti-(catalase-1) serum. Subcellular fractionation and indirect immunoelectron microscopy of the nematode detected catalase-2 inside vesicles judged to be peroxisomes using morphological criteria. The purified enzyme (220 kDa) was tetrameric, similar to many catalases from various sources, but exhibited unique pH optima for catalase (pH 6) and peroxidase (pH 4) activities; the latter value is unusually low and explains why the peroxidase activity was undetectable using the standard alkaline diaminobenzidine-staining method. These results indicate that catalase-2 is peroxisomal and verify that it can be used as a marker enzyme for C. elegans peroxisomes.

New aspects of sterol carrier protein 2 (nonspecific lipid-transfer protein) in fusion proteins and in peroxisomes.

Sterol carrier protein 2 (SCP2) is a 13-kDa peroxisomal protein, identical to nonspecific lipid-transfer protein, and stimulates various steps of cholesterol metabolism in vitro. Although the name is reminiscent of acyl carrier protein, which is involved in fatty acid synthesis, SCP2 does not bind to lipids specifically or stoichiometrically. This protein is expressed either as a small precursor or as a large fusion (termed SCPx) that carries at its C-terminal the complete sequence of SCP2. SCPx exhibits 3-oxoacyl-CoA thiolase activity, as well as sterol-carrier and lipid-transfer activities. The N- and C-terminal parts of SCPx are similar to the nematode protein P-44 and the yeast protein PXP-18, respectively. P-44, which has no SCP2 sequence, thiolytically cleaved the side chain of bile acid intermediate at a rate comparable to that of SCPx. This, together with the properties of other fusions with SCP2-like sequence, suggests that the SCP2 part of SCPx does not play a direct role in thiolase reaction. PXP-18, located predominantly inside peroxisomes, is similar to SCP2 in primary structure and lipid-transfer activity, and protects peroxisomal acyl-CoA oxidase from thermal denaturation. PXP-18 dimerized at a high temperature, formed an equimolar complex with the oxidase subunit, and released the active enzyme from the complex when the temperature went down. This article attempts to gain insight into the role of SCP2, and to present a model in which PXP-18, a member of the SCP2 family, functions as a molecular chaperone in peroxisomes.

Metabolic significance and expression of Caenorhabditis elegans type II 3-oxoacyl-CoA thiolase.

The authors cloned the cDNA of the nematode Caenorhabditis elegans encoding a 44-kDa protein (P-44), which is similar to sterol carrier protein x (SCPx). Genomic DNA data and Northern blot analysis excluded the possibility of P-44 forming SCPx-like fusion protein. P-44 is required in the formation of bile acid in vitro from CoA esters of their enoyl-form intermediate in the presence of D-3-hydroxyacyl-CoA dehydratase/D-3-dehydrogenase bifunctional protein. Also, rat SCPx converts 24-hydroxy-form intermediate to bile acid under similar conditions. From this and other evidence, P-44 and SCPx were categorized as type II thiolase. The mRNA encoding P-44 was detected in every developmental stage of C. elegans: egg, larval stages, and adult. P-44, therefore, seems essential for the normal functioning of this organism.

Type-II 3-oxoacyl-CoA thiolase of the nematode Caenorhabditis elegans is located in peroxisomes, highly expressed during larval stages and induced by clofibrate.

We examined the expression and localization of type-II 3-oxoacyl-CoA thiolase in the nematode Caenorhabditis elegans. Type-II thiolase acts on 3-oxoacyl-CoA esters with a methyl group at the alpha carbon, whereas conventional thiolases do not. Mammalian type-II thiolase, which is also termed sterol carrier protein x (SCPx) or SCP2/3-oxoacyl-CoA thiolase, is located in the peroxisomes and involved in phytanic acid degradation and most probably in bile acid synthesis. The nematode enzyme lacks the SCP2 domain, which carries the peroxisomal-targeting signal, but produces bile acids in a cell-free system. Northern and Western blot analyses demonstrated that C. elegans expressed type-II thiolase throughout its life cycle, especially during the larval stages, and that the expression was significantly enhanced by the addition of clofibrate at 5 mM or more to the culture medium. Whole-mount in situ hybridization and immunostaining of L4 larvae revealed that the enzyme was mainly expressed in intestinal cells, which are multifunctional like many of the cell types in C. elegans. Subcellular fractionation and indirect immunoelectron microscopy of the nematode detected the enzyme in the matrix of peroxisomes. These results indicate the fundamental homology between mammalian SCPx and the nematode enzyme regardless of whether the SCP2 part is fused, suggesting their common physiological roles.

An accurate adjustable applicator for magnetic resonance imaging-based stereotactic procedure using the Leksell G frame.

OBJECTIVE: An applicator system for the Leksell G frame was constructed to enable accurate placement of the frame for stereotactic magnetic resonance imaging (MRI) and successful stereotactic surgery. The applicator prevents inaccurate placement of the fiducial box on the patient's head and prevents contact of the frame holder with the patient's shoulder while in the MRI unit. It also helps to ensure optimal positioning of desired targets within the three-dimensional stereotactic space defined by the frame. METHODS: The applicator is made of transparent acrylic plates, which simulate the fiducial box that is attached to the frame for the preoperative stereotactic MRI study. An air cuff at the top supports the frame at any desired height and makes minute adjustments possible. Side cuffs help to keep the frame at the desired position from right to left. Indicators attached to the frame for the anterior fiducial plate prevent potential contact of the plate with the anterior posts and help avoid a poor fit caused by bending of the frame from excessive torque on the cranium fixation screws. Indicators for the MRI frame holder on the foot screws predict potential collision of the holder with the patient's shoulder before actually applying the holder on the frame. The applicator shows the range and limits of the Leksell stereotactic arc. RESULTS: This applicator system has been used effectively in more than 89 MRI-based functional stereotactic procedures. These include pallidotomy, thalamotomy, implantation of deep brain stimulators, and implantation of depth electrodes. It has functioned well and has facilitated excellent operative results in these cases. CONCLUSION: This simple frame applicator eliminates the need for reapplication of the stereotactic frame and additional imaging studies, thus providing successful and appropriate frame placement for stereotactic surgery.

Optimizing accuracy in magnetic resonance imaging-guided stereotaxis: a technique with validation based on the anterior commissure-posterior commissure line.

OBJECT: Some of the earliest successful frame-based stereotactic interventions directed toward the thalamus and basal ganglia depended on identifying the anterior commissure (AC) and posterior commissure (PC) in a sagittal ventriculogram and defining the intercommissural line that connects them in the midsagittal plane. The AC-PC line became the essential landmark for the localization of neuroanatomical targets in the basal ganglia and diencephalon and for relating them to stereotactic atlases. Stereotactic/functional neurosurgery has come to rely increasingly on magnetic resonance (MR) imaging guidance, and methods for accurately determining the AC-PC line on MR imaging are being developed. The goal of the present article is to present the authors' technique. METHODS: The technique described uses MR sequences that minimize geometric distortion and registration error, thereby maximizing accuracy in AC-PC line determinations from axially displayed MR data. The technique is based on the authors' experience with the Leksell G-frame but can be generalized to other MR imaging-based stereotactic systems. This methodology has been used in a series of 62 stereotactic procedures in 47 adults (55 pallidotomies and seven thalamotomies) with preliminary results that compare favorably with results reported when using microelectrode recordings. The measurements of the AC-PC line reported here also compare favorably with those based on ventriculography and computerized tomography scanning. CONCLUSIONS: The methodology reported here is critical in maintaining the accuracy and utility of MR imaging as its role in modern stereotaxy expands. Accurate parameters such as these aid in ensuring the safety, efficacy, and reproducibility of MR-guided stereotactic procedures.

A dissecting aneurysm of the posteroinferior cerebellar artery: case report.

OBJECTIVE AND IMPORTANCE: We present a patient who experienced a subarachnoid hemorrhage secondary to a dissecting aneurysm of the right posteroinferior cerebellar artery (PICA). The use of an encircling clip in treating the aneurysm while preserving supply to brain stem perforators originating near the dissecting segment and the distal PICA territory was key in the operative management. CLINICAL PRESENTATION: A 48-year-old patient with a history of hypertension presented with subarachnoid hemorrhage confirmed by computed tomography of the brain. Successive cerebral angiography revealed a dynamic change in the configuration of the dissection, with expansion of the associated focal ectasia. OPERATIVE MANAGEMENT: At surgery, three brain stem perforators adjacent to the aneurysm were visualized. The dissecting segment was reconstructed with an encircling Sundt clip and muslin wrap, which preserved the flow through the PICA and brain stem perforators. CONCLUSION: A patient suffering from a dissecting PICA aneurysm and subarachnoid hemorrhage was successfully treated with direct surgical reconstruction of the parent artery, sparing the perforators to the medulla.

Thiolase involved in bile acid formation.

The formation of cholic acid and chenodeoxycholic acid through cleavage of the side chains of CoA esters of 3alpha,7alpha,12alpha-trihydroxy-5beta-choles tan-26-oic acid and 3alpha,7alpha-dihydroxy-5beta-cholestan-26-oic acid is believed to occur in peroxisomes. Recently, we found a new peroxisomal enzyme, D-3-hydroxyacyl-CoA dehydratase/D-3-hydroxyacyl-CoA dehydrogenase bifunctional protein, and suggested that this bifunctional protein is responsible for the conversion of 3alpha,7alpha,12alpha-trihydroxy-5beta-cholest-2 4-en-26-oyl-CoA and 3alpha,7alpha-dihydroxy-5beta-cholest-24-en-26-oyl-CoA to their 24-oxo-forms. In the present study, the products of this bifunctional protein reaction were analyzed by gas chromatography-mass spectrometry, and the formation of 24-oxo-27-nor-cholestanes was confirmed. Previously, we found a new thiolase in Caenorhabditis elegans, P-44, and suggested that P-44 and sterol carrier protein x, a peroxisomal protein, constitute a second group of 3-oxoacyl-CoA thiolases. The production of cholic acid and chenodeoxycholic acid from the precursors on incubation with the bifunctional protein and sterol carrier protein x or P-44 was confirmed by gas chromatography.

A burr hole button to secure the electrode cable in depth electrode placement. Technical note.

A simple magnetic resonance imaging-compatible buttonlike device was devised to fix a depth electrode cable securely in the burr hole used for its insertion during surgery for depth electrode placement. The button is tightly fixed in the burr hole and it holds the cable without allowing protrusion or tension on the wound.

Identification and localization of intracerebral vessels by microvascular doppler in stereotactic pallidotomy and thalamotomy: technical note.

OBJECTIVE: To identify intracerebral vessels in proximity to the target for thermocoagulation in functional neurosurgery, we use a microvascular doppler sensor held in a special supporting needle that fits in the straightening cannula for the thermocoagulation needle. TECHNIQUE: After insertion of the straightening cannula aimed at the stereotactic target, the microvascular doppler probe positioned at the tip of a supporting hollow needle is advanced through the cannula. The proximal micrometer gauge indicates the depth of the tip of the doppler probe. By setting the doppler device to the shortest focusing depth (0.1 mm), the maximum pulsatile vascular sound indicates the depth of the vessel. RESULTS AND CONCLUSION: A prominent vascular sound was identified in 3 of 13 cases. By adjusting the depth of the target, no major bleeding was experienced after thermocoagulation lesions were made. This technique secures and protects the fragile microvascular doppler and identifies any significant arterial vessels at the stereotactic target, thus avoiding vascular injury.

A second isoform of 3-ketoacyl-CoA thiolase found in Caenorhabditis elegans, which is similar to sterol carrier protein x but lacks the sequence of sterol carrier protein 2.

We cloned a full-length cDNA of the nematode Caenorhabditis elegans that encodes a 44-kDa protein (P-44, 412 residues) similar to sterol carrier protein x (SCPx). Mammalian SCPx is a bipartite protein: its 404-residue N-terminal and 143-residue C-terminal domains are similar to 3-ketoacyl-CoA thiolase and identical to the precursor of sterol carrier protein 2 (SCP2; also termed non-specific lipid-transfer protein), respectively. P-44 has 56% sequence identity to the thiolase domain of SCPx but lacks the SCP2 sequence. Northern blot analysis revealed only a single mRNA species of 1.4 kb, which agrees well with the length of the cDNA (1371 bp), making it improbable that alternative splicing produces an SCPx-like fusion protein. The sequence similarities of P-44 to conventional thiolases are lesser than that to SCPx. Purified recombinant P-44 cleaved long-chain 3-ketoacyl-CoAs (C(8-16)) in a thiolytic manner by the ping-pong bi-bi reaction mechanism. The inhibition of P-44 by acetyl-CoA was competitive with CoA and non-competitive with 3-ketooctanoyl-CoA. This pattern of inhibition is shared with SCPx but not with conventional 3-ketoacyl-CoA thiolase, which is inhibited uncompetitively with respect to 3-ketoacyl-CoA. From these results, we concluded that nematode P-44 and mammalian SCPx constitute a second isoform of thiolase, which we propose to term type-II 3-ketoacyl-CoA thiolase.