O6 -methylguanine methyltransferase promoter methylation status of glioblastoma cell line clonal population.

Glioblastoma (GBM) remains a treatment-resistant malignant brain tumor in large part because of its genetic heterogeneity and epigenetic plasticity. In this study, we investigated the epigenetic heterogeneity of GBM by evaluating the methylation status of the O6 -methylguanine methyltransferase (MGMT) promoter in individual clones of a single cell derived from GBM cell lines. The U251 and U373 GBM cell lines, from the Brain Tumour Research Centre of the Montreal Neurological Institute, were used for the experiments. To evaluate the methylation status of the MGMT promoter, pyrosequencing and methylation-specific PCR (MSP) were used. Moreover, mRNA and protein expression levels of MGMT in the individual GBM clones were evaluated. The HeLa cell line, which hyper-expresses MGMT, was used as control. A total of 12 U251 and 12 U373 clones were isolated. The methylation status of 83 of 97 CpG sites in the MGMT promoter were evaluated by pyrosequencing, and 11 methylated CpG sites and 13 unmethylated CpG sites were evaluated by MSP. The methylation status by pyrosequencing was relatively high at CpG sites 3-8, 20-35, and 7-83, in both the U251 and U373 clones. Neither MGMT mRNA nor protein was detected in any clone. These findings demonstrate tumor heterogeneity among individual clones derived from a single GBM cell. MGMT expression may be regulated, not only by methylation of the MGMT promoter but by other factors as well. Further studies are needed to clarify the mechanisms underlying the epigenetic heterogeneity and plasticity of GBM.

Quantitative GABA magnetic resonance spectroscopy as a measure of motor learning function in the motor cortex after subarachnoid hemorrhage.

The neural mechanisms underlying gross and fine motor dysfunction after subarachnoid hemorrhage (SAH) remain unknown. The γ-aminobutyric acid (GABA) deficit hypothesis proposes that reduced neuronal GABA concentrations and the subsequent lack of GABA-mediated inhibition cause motor impairment after SAH. This study aimed to explore the correlation between GABA levels and a behavioral measure of motor performance in patients with SAH. Motor cortical GABA levels were assessed in 40 patients with SAH and 10 age-matched healthy controls using proton magnetic resonance spectroscopy. The GABA and N-acetylasparate (NAA) ratio was measured in the normal gray matter within the primary motor cortex. The relationship between GABA concentration and hand-motor performance was also evaluated. Results showed significantly lower GABA levels in patients with SAH's left motor cortex than in controls (GABA/NAA ratio: 0.282 ± 0.085 vs. 0.341 ± 0.031, respectively; p = 0.041). Reaction times (RTs), a behavioral measure of motor performance potentially dependent on GABAergic synaptic transmission, were significantly longer in patients than in controls (936.8 ± 303.8 vs. 440.2 ± 67.3 ms, respectively; p < 0.001). Moreover, motor cortical GABA levels and RTs exhibited a significant positive linear correlation among patients (r = 0.572, rs = 0.327, p = 0.0001). Therefore, a decrease in GABA levels in the primary motor cortex after SAH may lead to impaired cortical inhibition of neuronal function and indicates that GABA-mediated synaptic transmission in the motor cortex is critical for RT.

Monophasic-quadri-burst stimulation robustly activates bilateral swallowing motor cortices.

A stable, reliable, non-invasive, quantitative assessment of swallowing function remains to be established. Transcranial magnetic stimulation (TMS) is commonly used to aid in the diagnosis of dysphagia. Most diagnostic applications involve single-pulse TMS and motor evoked potential (MEP) recordings, the use of which is not clinically suitable in patients with severe dysphagia given the large variability in MEPs measured from the muscles involved in swallowing. Previously, we developed a TMS device that can deliver quadripulse theta-burst stimulation in 16 monophasic magnetic pulses through a single coil, enabling the measurement of MEPs related to hand function. We applied a system for MEP conditioning that relies on a 5 ms interval-monophasic quadripulse magnetic stimulation (QPS5) paradigm to produce 5 ms interval-four sets of four burst trains; quadri-burst stimulation (QBS5), which is expected to induce long-term potentiation (LTP) in the stroke patient motor cortex. Our analysis indicated that QBS5 conditioned left motor cortex induced robust facilitation in the bilateral mylohyoid MEPs. Swallowing dysfunction scores after intracerebral hemorrhage were significantly correlated with QBS5 conditioned-MEP parameters, including resting motor threshold and amplitude. The degree of bilateral mylohyoid MEP facilitation after left side motor cortical QBS5 conditioning and the grade of severity of swallowing dysfunction exhibited a significant linear correlation (r = -0.48/-0.46 and 0.83/0.83; R2 = 0.23/0.21 and 0.68/0.68, P < 0.001; Rt./Lt. side MEP-RMT and amplitudes, respectively). The present results indicate that RMT and amplitude of bilateral mylohyoid-MEPs after left motor cortical QBS5 conditioning as surrogate quantitative biomarkers for swallowing dysfunction after ICH. Thus, the safety and limitations of QBS5 conditioned-MEPs in this population should be further explored.

Partially thrombosed middle cerebral artery-lenticulostriate artery aneurysm with native radiological examinations suggesting proximal lenticulostriate artery aneurysm: A case report.

BACKGROUND: Preservation of the lenticulostriate artery (LSA) is crucial. LSAs usually cannot be spared with LSA aneurysms, when surgical clipping/excision or endovascular embolization of the LSA itself is performed. On the other hand, the LSA should be separated and preserved for proximal middle cerebral artery (M1)-LSA aneurysms. CASE DESCRIPTION: We report a case of M1-LSA aneurysm with native radiological examinations suggesting LSA aneurysm. The highlight of this unusual case was that during surgery, the aneurysm orifice was almost covered with thrombus and blood flow in an aneurysm that appeared separate from M1. Partial thrombectomy-clip reconstruction was performed, and M1 and LSAs were well preserved. CONCLUSION: Even with currently developed radiological modalities, thrombosed intracranial aneurysms may be misdiagnosed, depending on intraluminal flow conditions. Intraoperative findings from craniotomy sometimes contribute to a better understanding of the pathophysiology and decisions on appropriate treatment strategy.

Trauma may affect vasa vasorum to promote thrombosis and enlargement of intracranial aneurysms: A case report.

BACKGROUND: Thrombosed intracranial aneurysm (IA) is likely to occur in large or giant IAs. Almost all thrombosed IAs are found already in a thrombosed state, and few reports have depicted the process of thrombosis in unthrombosed aneurysm. Moreover, no reports appear to have described IA in which thrombosis accelerated after trauma. CASE DESCRIPTION: We report herein a case in which an unthrombosed large cerebral aneurysm rapidly thrombosed and grew within 3 months after trauma. The highlight in this unusual case was that during surgery, the aneurysm and anterior skull base were adherent and some blood vessels bridged between the aneurysm and dura mater. Histologically, intramural hemorrhage was seen in the tunica media of the aneurysm. CONCLUSION: Trauma may act as a "second hit" causing adhesion between IAs and surrounding tissues, accelerating inflammation of the vasa vasorum and aneurysmal walls, and thrombosis in IAs.

Cerebral Arteriovenous Malformations as Acquired Lesions: Case Reports and Review of the Literature.

Cerebral arteriovenous malformations (AVMs) are generally attributed to congenital lesions that arise from aberrant vasculogenesis between the fourth and eighth weeks of embryonic life. However, this dogma has been challenged by several recent observations, one of which is de novo formation of AVMs. Forty cases of de novo AVMs were published between 2000 and 2019, all of which involved a history of intracranial insult, such as vascular abnormalities or nonvascular conditions, prior to AVM diagnosis. We hereby present two unique operative cases of ruptured de novo AVMs in older adult patients. Case 1 is novel in the sense that the patient did not experience any kind of environmental trigger ("second hit") such as a previous intracranial insult, while Case 2 serves as the second report of a de novo AVM patient with a medical history of Bell's palsy. Although the exact mechanisms of AVM formation remain to be elucidated, it is likely to be a multifactorial process related to environmental and hemodynamic factors.

Lactate and Lactate Dehydrogenase in Cistern as Biomarkers of Early Brain Injury and Delayed Cerebral Ischemia of Subarachnoid Hemorrhage.

OBJECTIVE: The pathophysiology of delayed cerebral ischemia (DCI) following aneurysmal subarachnoid hemorrhage (aSAH) has not been fully evaluated. The aim of this study was to evaluate the dynamics of lactate and lactate dehydrogenase (LDH) in carotid cisternal cerebrospinal fluid (CSF), and to discuss their effectiveness as markers of early brain injury (EBI) and DCI following aSAH. PATIENTS AND METHODS: Among 91 consecutive aSAH patients treated between January 2012 and March 2019 at National Hospital Organization Beppu Medical Center, 19 patients (20.9%) were eligible for this retrospective study. Concentrations of lactate and LDH in carotid cisternal CSF within 14 days after onset of aSAH were evaluated. RESULTS: Six of the 19 patients (31.6%) had a history of DCI. Both lactate and LDH levels in carotid cisternal CSF were significantly higher in the DCI group than in the non-DCI group on postbleeding day (PBD) 1-2, 3-4, and 5-6. Interestingly, neither lactate nor LDH levels in blood differed significantly between DCI and non-DCI groups on PBD 1-2. CONCLUSIONS: Lactate and LDH concentrations in carotid cisternal CSF may vividly reflect the EBI and may thus represent predictive biomarkers of DCI following aSAH.

The Potentiality for Development of Multiple Dural Arteriovenous Fistulas after Ligation of the Internal Jugular Vein: A Case Report.

A 74-year-old male presented with an intracranial hemorrhage caused by multiple dural arteriovenous fistulas (DAVFs) in the left transverse sinus and right sigmoid sinus. Four months previously, the patient underwent tongue cancer removal with lymph node dissection and ligation of the right internal jugular vein. Endovascular embolization (transvenous and transarterial embolization) resulted in the complete disappearance of the fistulas. Follow-up angiography revealed new arteriovenous shunts at the superior sagittal sinus and right transverse sinus, and we treated the patient with staged transarterial embolization. Finally, venous congestion almost completely resolved and the DAVFs disappeared without any sign of recurrence. This case speculates the concept of DAVF as an acquired lesion caused by intravenous hypertension and alerts clinicians to take precautions against ligation of the internal jugular vein during a cervical operation.

Third nerve palsy caused by compression of the posterior communicating artery aneurysm does not depend on the size of the aneurysm, but on the distance between the ICA and the anterior-posterior clinoid process.

OBJECTIVE: Third nerve palsy (TNP) caused by a posterior communicating artery (PCoA) aneurysm is a well-known symptom of the condition, but the characteristics of unruptured PCoA aneurysm-associated third nerve palsy have not been fully evaluated. The aim of this study was to analyze the anatomical features of PCoA aneurysms that caused TNP from the viewpoint of the relationship between the ICA and the skull base. METHODS: Forty-eight unruptured PCoA aneurysms were treated surgically between January 2008 and September 2013. The characteristics of the aneurysms were evaluated. RESULTS: Thirteen of the 48 patients (27%) had a history of TNP. The distance between the ICA and the anterior-posterior clinoid process (ICA-APC distance) was significantly shorter in the TNP group (p<0.01), but the maximum size of the aneurysms was not (p=0.534). CONCLUSION: Relatively small unruptured PCoA aneurysms can cause third nerve palsy if the ICA runs close to the skull base.

Increasing high-frequency oscillations (HFOs) in patients with brain tumours: implication for increasing amplitude of N20.

OBJECTIVE: The N20 and high-frequency oscillations (HFOs) of somatosensory evoked potentials (SEPs) were recorded in patients with brain tumours. This study sought to estimate how a brain tumour could increase the peak amplitude of N20, while also illustrating the clinical significance of this condition. METHODS: Median nerve SEPs were recorded in 34 conscious patients, who were admitted to the hospital owing to the presence of a circumscribed unilateral brain tumour. Eleven patients showed an increasing peak amplitude of N20 on the affected side (AS). HFOs were used to analyse the underlying mechanism. RESULTS: While the amplitude of N20 in AS was higher than that on the normal side (NS), the latency of N20 showed no difference on either side. The amplitude of the early components of HFOs on the AS was higher than that on the NS (p=0.015), but the latency was not significantly different. The amplitude of late HFOs on the AS was also higher than on the NS (p=0.041), and the latency was also not significantly different. Our findings proved an increasing amplitude of HFOs to be a discrete character in AS>NS group, thereby indicating that a sensory disturbance was not commonly expressed in AS>NS groups. CONCLUSIONS: These results suggested that the hyperexcitability in the thalamocortical pathway were responsible for this condition. Hyperexcitability was presumably caused by the influence of the corticothalamic feedback and the neural interactions between the relay neurons and the reticular neurons. The clinically significant finding was that an increasing amplitude of N20 thus indicated the presence of a sub-clinical change. SIGNIFICANCE: A brain tumour could increase the amplitude of N20 due to the hyperexcitability in the thalamocortical pathway. An increasing amplitude of N20 thus indicated the presence of a sub-clinical change in the thalamocortical pathway on the side of the tumour.

Superficial temporal artery-to-middle cerebral artery anastomosis with encephalo-duro-myo-synangiosis as a modified operative procedure for moyamoya disease.

BACKGROUND: Various types of revascularization surgery have been performed for moyamoya disease. Although the efficacies of these operations are well recognized, the optimal operative procedure remains undecided. In this report, we describe our modified surgical revascularization procedure for moyamoya disease and retrospectively analyze the results of such surgeries on six sides in six adult patients. METHODS: Our operative procedure, combining direct and indirect bypasses, is a superficial temporal artery to middle cerebral artery anastomosis with encephalo-duro-myo-synangiosis. The encephalo-duro-myo-synangiosis is an indirect bypass combining the encephalo-duro- and encephalo-myo-synangioses. This operative procedure has been used routinely in adult patients since 2002. RESULTS: Perioperative complications were noted in one of the six operations. This complication was transient and no attributive lesions were detected on CT or MRI. Revascularization was seen in cerebral blood flow studies in all patients, and the clinical outcomes were excellent or good. Effective neovascularization through the grafts was observed in all patients in follow-up angiographies. CONCLUSIONS: This operative procedure provides needed revascularization and prevents ischemic deficits. This modified procedure is useful for responding to subsequent additional ischemia in the area of the anterior cerebral artery and should be considered one of the optimal procedures for treating moyamoya disease.

Malignant fibrous histiocytosis after high-dose proton radiation therapy for anaplastic astrocytoma.

This report presents a 70-year-old male who presented with a rare malignant fibrous histiocytosis after high-dose proton radiation therapy for anaplastic astrocytoma. To our knowledge, malignant fibrous histiocytosis caused by proton therapy has not been reported, therefore the clinical features of this complication are described and previous cases are reviewed.

A hemispherotomy for intractable startle epilepsy characterized by infantile hemiplegia and drop attacks.

Startle epilepsy is provoked by unexpected sensory stimuli, mainly auditory, and reveals subsequent tonic posturing of the limbs. We present a case of intractable startle epilepsy with infantile hemiplegia and discuss the indications for a hemispherotomy.

Functional motor recovery of an infant after a huge ependymoma resection.

Huge supratentorial ependymomas are rarely encountered tumors, even in the infant population. A recovery from complete hemiplegia following a tumor resection including the primary motor cortex was observed. A 5-month-old girl presented with a conjugate deviation to the right and a head circumference that had gradually expanded since birth. Magnetic resonance imaging (MRI) demonstrated a well-enhanced huge mass extending into the right hemisphere. A subtotal removal with the primary motor cortex was performed. However, a regrowth of the residual tumor was observed and, thereafter, the patient underwent a subsequent surgical intervention 5 months later. The histological findings demonstrated an ependymoma. Her motor function was dramatically improved after rehabilitation and no tumor recurrence was detected for 10 years. A diffusion tensor imaging study showed that the motor fibers arose from the residual frontal lobe. The successful surgical management of ependymoma may depend on a total microscopic resection. In a case demonstrating a huge ependymoma, we had to remove a very thin motor cortex with the tumor. However, the motor function recovered completely. The motor damage inflicted at an early developmental age may be fully compensated due to the neuroplasticity of the residual brain.

A rosette-forming glioneuronal tumor of the spinal cord: the first case of a rosette-forming glioneuronal tumor originating from the spinal cord.

Rosette-forming glioneuronal tumors of the fourth ventricle are rare brain tumors, and only 19 such lesions have been previously reported. This report presents the first case of a rosette-forming glioneuronal tumors arising from the spinal cord. A 44-year-old woman presented with a 15-year history of dissociated sensory disturbance of the lower extremities that gradually spread through her upper extremities. She also experienced continuing motor disturbance. Magnetic resonance imaging demonstrated a mass in the cervicothoracic spinal cord that suggested an intramedullary spinal tumor. A total gross resection of the tumor was performed. As is typical of rosette-forming glioneuronal tumors of the fourth ventricle, this spinal cord example manifested neurocytic and astrocytic components. Neurocytic rosettes were detected in the neurocytic component, and the center of rosettes showed positive immunostaining for synaptophysin. The astrocytic component showed characteristic features of a pilocytic astrocytoma, as is often the case in the fourth ventricle examples.

Induction of collateral circulation by hypoxia-inducible factor 1alpha decreased cerebral infarction in the rat.

BACKGROUND: We recently reported that hypoxia-inducible factor 1alpha (HIF-1alpha), HIF-2alpha and cyclooxygenase 2 naked DNA induced angiogenesis in a rat indirect bypass model. In this work, we investigated whether the collateral circulation induced by HIF-1alpha DNA affected the cerebral infarction. METHODS: We utilized a rat encephalomyosynangiosis (EMS) model and inoculated HIF-1alpha DNA onto the brain surface. These treatments were performed before the cerebral infarction occurred. We thereafter performed middle cerebral artery occlusion on the fifth or tenth day after EMS. RESULTS: A histological section treated with HIF-1alpha DNA for 10 days showed a well-developed collateral circulation (p<0.05) and a reduction in the infarction volume in comparison to the control DNA (p<0.01). CONCLUSION: These results suggest the feasibility of a novel approach for the treatment of cerebral ischemia via the development of therapeutic collateral circulation, in which neovascularization may be indirectly achieved using a transcriptional regulatory strategy.

Induced angiogenesis under cerebral ischemia by cyclooxygenase 2 and hypoxia-inducible factor naked DNA in a rat indirect-bypass model.

We recently reported that hypoxic stress induces the expression of HIF-1alpha, HIF-2alpha, cyclooxygenases-2 (COX-2) and VEGF in vivo. In this study, we investigated whether HIF-1alpha, HIF-2alpha, or COX-2 naked DNA induced angiogenesis in a cerebral ischemic model in vivo. We utilized a rat encephalo-myo-synangiosis (EMS) model and inoculated naked DNA into the brain surface. We analyzed whether DNA induced angiogenic factors and neovascularization. New blood vessel formation was detected by anti-Factor VIII staining. A histological section treated with HIF-2alpha or COX-2 DNA showed an increased expression of VEGF with angiogenesis, in comparison to the control DNA. The HIF-1alpha, HIF-2alpha, and COX-2 are able to promote significant angiogenesis development. These results suggest the feasibility of a novel approach for therapeutic angiogenesis of cerebral ischemia in which neovascularization may be indirectly achieved using a transcriptional and cytokine's regulatory strategy.

Postoperative adjuvant treatment for pineal parenchymal tumour of intermediate differentiation.

Pineal parenchymal tumour of intermediate differentiation (PPTID) in adults is rare and a treatment strategy for this condition has not yet been established. We present a case of an elderly patient treated with postoperative adjuvant therapy using radio- and chemotherapy. This 60-year-old man presented with a 3-month history of memory disturbance, gait instability and double vision. Computed tomography and magnetic resonance imaging demonstrated a mass in the pineal region that suggested a malignant tumour. Partial removal of the tumour was undertaken via the right occipital transtentorial approach. The histological diagnosis was PPTID. Postoperative radio- and chemotherapy were administered, with a good response. Little is known about the clinical behaviour of PPTID in adults. Our treatment plan indicates one effective option for the management of such tumours.