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PDF file - 102K, This figure demonstrates the validities of knockdowns of RASSF3 (in HeLa, U2OS, and MCF-7 cells), p53 (in HeLa and U2OS cells), and MDM2 (in HeLa and U2OS cells)

PDF file - 135K, This figure demonstrates that RASSF3 interacts with MST2 but that RASSF3-induced apoptosis in HCT116 cells is independent of the Hippo pathway but is dependent on MOAP1

PDF file - 148K, This figure demonstrates that RASSF3 regulates cell cycle in HeLa cells and that RASSF3 is involved in UVB- and VP-16-induced G1/S arrest in HeLa cells

RASSF3 is the smallest member of the RASSF family of proteins that function as tumor suppressors. Unlike other members of this important family, the mechanisms through which RASSF3 suppresses tumor formation remain unknown. Here, we show that RASSF3 expression induces p53-dependent apoptosis and its depletion attenuates DNA damage–induced apoptosis. We found that RASSF3-induced apoptosis depended upon p53 expression. Exogenous expression of RASSF3 induced G1–S arrest, which was also p53 dependent. In contrast, loss of RASSF3 promoted cell-cycle progression, abrogated UVB- and VP-16–induced G1–S arrest, decreased p53 protein and target gene expression, and prevented DNA repair. RASSF3 was shown to directly interact with and facilitate the ubiquitination of MDM2, the E3 ligase that targets p53 for degradation, thereby increasing p53 stabilization. Together, our findings show the tumor suppressor activity of RASSF3, which occurs through p53 stabilization and regulation of apoptosis and the cell cycle. Cancer Res; 72(11); 2901–11. ©2012 AACR.

PDF file - 151K, This figure demonstrates that RASSF3 is involved in DNA repair after UVB-induced DNA damage in U2OS cells

PDF file - 295K, This figure demonstrates that RASSF3 induces apoptosis in HeLa cells

PDF file - 163K, This figure demonstrates that RASSF3 suppresses p53 degradation in HeLa cells and that RASSF3 depletion causes polyploidy in HeLa cells

PDF file - 34K

PDF file - 137K, This figure demonstrates that RASSF3-induced apoptosis in HeLa cells is p53-dependent

PDF file - 19K, Sup Table 1: This table summarizes the primers for quantitative RT-PCR used in this study. Sup Table 2: This table summarizes the twenty-one-oligomers to knock down various genes used in this study

BACKGROUNDS: On March 11(th) 2011, the Tohoku region of Japan was struck by catastrophic disasters. Thousands of people were killed due to a magnitude 9.0 earthquake and its subsequent tsunami. Furthermore, a serious nuclear crisis occurred in Fukushima Prefecture as a result of the disasters, and an emergency evacuation was ordered to people living near the nuclear power plants. There was a lot of anxiety regarding lost families as well as the influences of radioactivity on the health of people and their children. Based on these urgent and uncertain situations, a number of research projects were developed at many institutes both inside and outside Fukushima. METHODS: We herein report the investigative research projects related to the Tohoku Earthquake (The Great East Japan Earthquake) conducted after the disasters. The research projects were reviewed by the Institutional Review Board in Fukushima Medical University during the two years following the disasters. The research projects conducted in universities other than Fukushima Medical University were also examined using questionnaire analysis. RESULTS: Among the research projects conducted in Fukushima Medical University (n=424), 7% (n=32) were disaster-related investigative research. The mean duration planned to pursue the projects was 25.5 months. Among these projects, those focusing on the health of Fukushima citizens were most common (n=9), followed by the influence of chronic exposure of radiation on chronic inflammatory disorders (n=6), and the mental health of Fukushima citizens (n=5). They were carefully reviewed for the purpose, suitability, and necessity from ethical as well as scientific viewpoints. The majority of the research projects focused on the effects of the Tohoku Earthquake and/or chronic exposure to low-dose radioactivity on the health of children and pregnant women, as well as on various disorders, such as mental health and chronic inflammatory diseases. On the other hand, among 58 projects we collected from 22 institutes in prefectures other than Fukushima, mental health-related projects were the most common (n=18), followed by radiation exposure-related projects (n=10). CONCLUSIONS: Many of these projects are ongoing, and in particular, long term follow-up regarding the health of the residents of Fukushima Prefecture, especially children and pregnant women, is necessary. Language: en

The aim of this study was to analyze the prognostic factors of trigeminal neuralgia (TN) after microvascular decompression (MVD), and to evaluate the volumetric parameters of the cerebellopontine angle (CPA) cistern as a pathogenic factor and imaging predictor. This retrospective study included 70 patients with primary TN treated with pure MVD, followed up for at least 1 year and evaluated by high-resolution MR imaging. The volume of the CPA cistern was calculated bilaterally, and the “Cistern Deviation Index” was defined to represent degree of deviation of the CPA cistern. Clinical data and volumetric parameters were compared between patients with TN and age- and sex-matched controls without TN, and between the recurrent and non-recurrent patients. The transposition procedure had a better outcome than the interposition procedure (P

Background Precise evaluation of hemodynamic stress is important for the treatment of moyamoya disease (MMD). Objective To explore whether dynamic susceptibility contrast magnetic resonance imaging could predict the effects and risk of indirect bypass surgery on MMD. Methods Clinical data of patients with MMD who were evaluated preoperatively and postoperatively with dynamic susceptibility contrast magnetic resonance imaging and digital subtraction angiography were evaluated retrospectively. Indirect bypass surgery was performed on 115 hemispheres of 69 patients (mean age, 15 years; range, 3-54 years). We examined the correlations of ischemic events and revascularization with the mean transit time (MTT) delay to cerebellum. Results The hemispheres that caused the ischemic events (responsible hemisphere) had a significantly longer preoperative MTT delay than the nonresponsible hemispheres (2.66 ± 1.34 vs 1.57 ± 1.09 seconds). The postoperative MTT delay fell significantly in the patients whose symptoms disappeared (preoperative, 2.61 ± 1.35 seconds; postoperative, 1.35 ± 0.96 seconds). Perioperative infarction occurred in 4 hemispheres (3.5%), and the MTT delay was significantly longer in those hemispheres than in the others (3.97 ± 1.20 vs 2.38 ± 1.34 seconds). The MTT delay was significantly longer in patients with higher angiographic stages. Indirect bypass surgery ameliorated the MTT delay to the same degree in adults and children. Digital subtraction angiography revealed that the induced revascularization was far superior in areas with longer MTT delays. Conclusion Dynamic susceptibility contrast magnetic resonance imaging proved to be a useful clinical imaging method for patients with MMD. It may be helpful for selecting candidates for MMD intervention and for predicting the effects and risks of surgery. Abbreviations DSC-MRI, dynamic susceptibility contrast magnetic resonance imagingMMD, moyamoya diseaseMTT, mean transit timeROI, region of interest.

Ruptured intracranial aneurysms are rare in the pediatric population compared to adults. This has incited considerable discussion on how to treat children with this condition. Here, we report a child with a ruptured saccular basilar artery aneurysm that was successfully treated with coil embolization. A 12-year-old boy with acute lymphoblastic leukemia and accompanying abdominal candidiasis after chemotherapy suddenly complained of a severe headache and suffered consciousness disturbance moments later. Computed tomography scans and cerebral angiography demonstrated acute hydrocephalus and subarachnoid hemorrhage caused by saccular basilar artery aneurysm rupture. External ventricular drainage was performed immediately. Because the patient was in severe condition and did not show remarkable signs of central nervous system infection in cerebrospinal fluid studies, we applied endovascular treatment for the ruptured saccular basilar artery aneurysm, which was successfully occluded with coils. The patient recovered without new neurological deficits after ventriculoperitoneal shunting. Recent reports indicate that both endovascular and microsurgical techniques can be used to effectively treat ruptured cerebral aneurysms in pediatric patients. A minimally invasive endovascular treatment was effective in the present case, but long-term follow-up will be necessary to confirm the efficiency of endovascular treatment for children with ruptured saccular basilar artery aneurysms.

Object Recent evidence suggests that a glioma stem cell subpopulation may determine the biological behavior of tumors, including resistance to therapy. To investigate this hypothesis, the authors examined varying grades of gliomas for stem cell marker expressions and histopathological changes between primary and recurrent tumors. Methods Tumor samples were collected during surgery from 70 patients with varying grades of gliomas (Grade II in 12 patients, Grade III in 16, and Grade IV in 42) prior to any adjuvant treatment. The samples were subjected to immunohistochemistry for MIB-1, factor VIII, GFAP, and stem cell markers (CD133 and nestin). Histopathological changes were compared between primary and recurrent tumors in 31 patients after radiation treatment and chemotherapy, including high-dose irradiation with additional stereotactic radiosurgery. Results CD133 expression on glioma cells was confined to de novo glioblastomas but was not observed in lower-grade gliomas. In de novo glioblastomas, the mean percentage of CD133-positive glioma cells in sections obtained at recurrence was 12.2% ± 10.3%, which was significantly higher than that obtained at the primary surgery (1.08% ± 1.78%). CD133 and Ki 67 dual-positive glioma cells were significantly increased in recurrent de novo glioblastomas as compared with those in primary tumors (14.5% ± 6.67% vs 2.16% ± 2.60%, respectively). In contrast, secondary glioblastomas rarely expressed CD133 antigen even after malignant progression following radiotherapy and chemotherapy. Conclusions The authors' results indicate that CD133-positive glioma stem cells could survive, change to a proliferative cancer stem cell phenotype, and cause recurrence in cases with de novo glioblastomas after radiotherapy and chemotherapy.

Susceptibility artifacts generated in magnetic resonance (MR) images were quantitatively evaluated for various metals using a three-dimensional (3-D) artifact rendering to demonstrate the correlation between magnetic susceptibility and artifact volume. Ten metals (stainless steel, Co-Cr alloy, Nb, Ti, Zr, Mo, Al, Sn, Cu and Ag) were prepared, and their magnetic susceptibilities measured using a magnetic balance. Each metal was embedded in a Ni-doped agarose gel phantom and the MR images of the metal-containing phantoms were taken using 1.5 and 3.0 T MR scanners under both fast spin echo and gradient echo conditions. 3-D renderings of the artifacts were constructed from the images and the artifact volumes were calculated for each metal. The artifact volumes of metals decreased with decreasing magnetic susceptibility, with the exception of Ag. Although Sn possesses the lowest absolute magnetic susceptibility (1.8×10(-6)), the artifact volume from Cu (-7.8×10(-6)) was smaller than that of Sn. This is because the magnetic susceptibility of Cu was close to that of the agarose gel phantom (-7.3×10(-6)). Since the difference in magnetic susceptibility between the agarose and Sn is close to that between the agarose and Ag (-17.5×10(-6)), their artifact volumes were almost the same, although they formed artifacts that were reversed in all three dimensions.

Object The authors compared the clinical features between familial and sporadic cases of moyamoya disease (MMD) by retrospectively analyzing data on patients with MMD registered in the database of Tokyo Medical and Dental University over a period of 28 years. Methods In total, 383 patients with hospital records at Tokyo Medical and Dental University from 1980 to 2007 were registered into the database. The data on all of these patients were retrospectively reviewed to clarify the occurrence of familial cases. Clinical features of child or adolescent patients (< 20 years of age) with MMD were compared between familial and sporadic cases in a subgroup of patients who were registered after 1995, initially diagnosed using MR angiography, and assessed using an intelligence scale. Results Familial occurrence was observed in 59 patients (15.4%) in 40 pedigrees. The clinical features of juvenile patients were analyzed in 124 patients, 22 (17.7%) of whom had familial histories. In comparison with the sporadic cases, patients with familial histories were significantly younger at onset (4.7 vs 6.6 years old), had significantly more cortical infarction (59.1% vs 25.5%), and had significantly more stenoocclusive lesions in the posterior cerebral artery (45.4% vs 24.5%). The rate of patients with intellectual disturbance (intelligence quotient < 75) was significantly larger in the familial cases (47.4%) than in the sporadic cases (17.8%). Conclusions This survey of the clinical features of familial MMD suggests that patients with familial MMD had a more serious clinical course in childhood than the sporadic MMD cases.

Object Surgical revascularization is considered an effective treatment for juvenile patients with moyamoya disease (MMD). Yet the long-term outcome in surgically treated patients still needs to be clarified. More than 30 years have passed since the authors' department started intensively treating pediatric patients with MMD using indirect anastomosis techniques. In this study the authors surveyed the current status of these patients. Methods Activities of daily living (ADLs) were surveyed and present clinical status was assessed based on the modified Rankin Scale (mRS). Cerebrovascular events subsequent to surgical treatment were also recorded. Results Since 1979, 208 patients younger than 19 years of age with MMD were surgically treated and followed up for > 3 years. Data were available on 172 patients (83%), who had been followed up for a mean of 14.3 years (range 3–32 years). Activity of daily living outcomes were as follows: 138 patients (80.2%) had mRS scores of 0–2, 29 (16.9%) a score of 3, 1 (0.6%) a score of 4, 1 (0.6%) a score of 5, and 3 (1.7%) a score of 6. Cerebrovascular events occurred 8 or more years after surgery in 6 patients (3.4%), that is, 6 hemorrhages and 3 infarctions. The cumulative risk of late-onset stroke at 10, 20, and 30 years after surgical intervention was 0.8%, 6.3%, and 10.0%, respectively. Conclusions This long-term survey demonstrated that most surgically treated pediatric patients with MMD maintain good ADL outcomes. However, a significant number of new cerebrovascular events occurred more than 10 years after the initial surgery. Additional follow-up will help to identify which events may occur during the adult years of patients treated as children.

Interest has continued in the neural substrates of language switch, which allows multilingual people to select an appropriate language. Using functional magnetic resonance imaging, we investigated the neural substrates for switching between different languages (cross-language switch) in comparison with those for switching between different tasks or between different action sequences. Subjects were 20 native Japanese (L1) speakers with moderate to high proficiency in English (L2). They were asked to judge pronunciation of visually presented Arabic numerals in either L1 or L2 (phonological judgment task) or the numerical meaning of the same stimuli (numerical judgment task). The switching of the tasks was semi-randomly cued by a background color change. Several brain regions showed significantly greater activity for the forward cross-language switching (L1 to L2) than the backward cross-language switching (L2 to L1). Such cross-language switch regions included the right prefrontal cortex (PFC), left superior temporal/supramarginal gyrus (STG/SMG), anterior cingulate cortex (ACC), left inferior frontal gyrus (IFG), and caudate nucleus. Among these cross-language switch regions, the left IFG and caudate nucleus were also involved in the task switching (switching between the phonological and numerical tasks). These findings were supported by an action-sequence switch experiment examining brain activity during switching among different tapping sequences. In the right PFC, moreover, activity during the forward cross-language switching was positively correlated with the subjects’ proficiency in L2. The present study suggests that the right PFC, left IFG, left STG/SMG, ACC, and caudate nucleus might subserve differential aspects of cross-language switch in late bilinguals.

We applied voltage-sensitive dye imaging to neonatal rat cortical slice preparations and analyzed developmental changes in synaptic plasticity, long-term potentiation (LTP), in the corticostriatal projection. Coronal slice preparations were dissected from postnatal 1- to 21-day (P1–P21) rats, and the transmembrane voltage-related optical signals evoked by cortical stimulation were recorded using a 464ch optical recording system with the voltage-sensitive absorption dye. In the striatum, the optical signal was composed of a fast spike-like signal followed by a slow signal, which corresponded to an action potential and an excitatory postsynaptic potential (EPSP), respectively. The slow signal could be detected at the P1 stage, suggesting that the EPSP is already expressed in the corticostriatal projection at least at early stages after birth. On the other hand, the slow signal was potentiated with a single shot of tetanic stimulation and the potentiation lasted at least 1 h, which is considered to correspond to long-term potentiation. With ontogenetic examinations, we found that (1) the EPSP could be potentiated with tetanic stimulation from the P9 stage and that (2) after the LTP induction, the potentiation was maintained for a longer time in the postnatal 3W stage than in the 2W stage. These results suggest that characteristics of LTP change dynamically during postnatal development.

The outcomes of postoperative antiepileptic drug (AED) cessation after temporal lobe epilepsy surgery were investigated to identify the preoperative factors associated with seizure-free status without drug treatment in 40 patients who underwent resective epilepsy surgery between 2000 and 2007 for the treatment of complex partial seizures of the type typically associated with temporal lobe epilepsy. All patients were followed up for at least 3.5 years after surgery. After the numbers and types of AEDs used as preoperative and postoperative treatments were determined, the patients who obtained seizure-free outcome for at least 2 years after AED cessation were analyzed. Sex, age at surgery, age at seizure onset, time to operation, seizure frequency, operation side, invasive study, pathological types (hippocampal sclerosis or others), history of generalized convulsive seizures, and number of preoperative AEDs were analyzed as factors potentially associated with seizure-free status without drug treatment. In total, 35% of the patients who underwent temporal lobe epilepsy surgery achieved seizure-free status without drugs. The mean number AEDs fell from 2.1 preoperatively to 1.2 after the surgery. Seizures recurred in 18% of the patients after AED withdrawal. A shorter duration from seizure onset to surgery was found to be significantly related to successful withdrawal of postoperative AED treatment. Our results suggested that early surgical intervention after seizure onset is an important precondition for achieving seizure-free status without drugs after temporal lobe epilepsy surgery.

Arterial spin labeling (ASL) and dynamic susceptibility contrast (DSC) magnetic resonance imaging (MRI) are widely used to image cerebral blood flow (CBF) in stroke. This study examined how changes in tissue spin-lattice relaxation-time constant ( T1), blood-brain barrier (BBB) permeability, and transit time affect CBF quantification by ASL and DSC in postischemic hyperperfusion in the same animals. In Group I ( n = 6), embolic stroke rats imaged 48hours after stroke showed regional hyperperfusion. In normal pixels, ASL- and DSC-CBF linearly correlated pixel-by-pixel. In hyperperfusion pixels, ASL-CBF was significantly higher than DSC-CBF pixel-by-pixel (by 25%). T1 increased from 1.76 ± 0.14 seconds in normal pixels to 1.93 ± 0.17 seconds in hyperperfusion pixels. Arterial transit time decreased from 300 milliseconds in normal pixels to 200 milliseconds in hyperperfusion pixels. Δ R2* profiles showed contrast-agent leakages in the hyperperfusion regions. In Group II ( n = 3) in which hypercapnic inhalation was used to increase CBF without BBB disruption, CBF increased overall but ASL- and DSC-CBF remained linearly correlated. In Group III ( n = 3) in which mannitol was used to break the BBB, ASL-CBF was significantly higher than DSC-CBF. We concluded that in normal tissue, ASL and DSC provide comparable quantitative CBF, whereas in postischemic hyperperfusion, ASL-CBF and DSC-CBF differed significantly because ischemia-induced changes in T1 and BBB permeability affected the two methods differently.

Background Moyamoya disease is one of the causes of higher brain dysfunction in younger patients. Fortunately, it may be possible to protect younger Moyamoya disease patients from brain dysfunction via surgical manoeuvres. Aim Our group retrospectively analysed the correlation between preoperative positron emission tomography data and the intelligence quotient scores of 60 Japanese Moyamoya disease patients (age range 9–64). Method All patients underwent a quantitative measurement of the cerebral metabolic rate of oxygen by inhalation of C15O2 and 15O2 gas with positron emission tomography. The data was analysed using SPM99 software to determine the cerebral regions in which regional cerebral metabolic rate of oxygen was significantly correlated with full-scale intelligence quotient, verbal intelligence quotient, or performance intelligence quotient measured using the Wechsler intelligence scale. Results All scores (full-scale intelligence quotient, verbal intelligence quotient, and performance intelligence quotient) showed significant positive correlations with the cerebral metabolic rate of oxygen in the lower part of the bilateral frontal lobe, the right anterior temporal lobe, and the medial occipital lobe. The verbal intelligence quotient was significantly and positively correlated with the cerebral metabolic rate of oxygen in the left inferior frontal lobe, including Broca's area. Infarcted lesions in the left posterior temporal lobe and the right upper frontal lobe influenced the decline of all of the intelligence quotient scores measured. Conclusion The present analysis indicates that the higher brain function of Moyamoya disease patients tends to be affected by the cerebral metabolism of specific regions. This information may be useful in seeking optimal clinical management to preserve higher brain function in patients with Moyamoya disease.

Whether the intentional antihypertensive therapy recommended by the American Heart Association/American Stroke Association (AHA/ASA) guidelines has clinical benefit for patients who have acute spontaneous intracerebral hemorrhage (ICH) has yet to be proven. We retrospectively reviewed the clinical charts of 175 patients with putaminal or thalamic ICH with acute hypertension to examine the correlation between the efficacy of antihypertensive therapy within 3 hours of onset, hematoma expansion (HE) after hospitalization and clinical outcome. The aim of the antihypertensive therapy was to achieve and maintain a systolic blood pressure of 120 mm Hg to 160 mm Hg until the second CT scan. The mean arterial pressure (MAP) after admission was the average MAP values measured every hour for the first 3 hours of hospitalization or until the second CT scan, if this was performed within the same timeframe. Thirty-two (18.3%) patients were found to have HE. Prior to the second CT scan, antihypertensive medications were administered to all patients without any major complications. A multiple logistic regression analysis revealed that a MAP of >110 mm Hg after admission was the only variable independently associated with HE (odds ratio [OR] = 3.455; 95% confidence interval [CI] = 1.510-8.412; p = 0.004). Modified Rankin Scale scores of < or = 3 on day 30 were significantly more common in those patients without HE (p = 0.002). Our findings suggest that there are clinical benefits, by the prevention of subsequent HE, in maintaining a MAP level lower than that recommended by the AHA/ASA (110 mm Hg) after hospitalization for patients who have ICH.

Traumatic brain injury (TBI) is one of the most acute degenerative pathologies in the central nervous system, and in vivo indices enabling an assessment of TBI on a mechanistic basis have yet to be established. The aim of this work was to pursue neuroinflammatory changes and their link to functional disruptions of traumatically-damaged neurons in a rat model of TBI by longitudinal positron emission tomographic (PET) assays. TBI was induced in the unilateral frontal cortex of craniotomied rats according to a lateral fluid percussion brain injury protocol. The use of [(18)F]fluoroethyl-DAA1106 as a PET tracer for translocator protein (TSPO) permitted demonstration of the inflammatory response to the injury, peaking at 1 week after impact. This alteration was parallel to metabolic deficits assessed by PET with [(18)F]fluorodeoxyglucose, but the difference in TSPO levels between impacted and non-impacted frontal cortices was more than threefold of the interlateral metabolic difference, indicating superiority of TSPO imaging for sensitive detection of post-traumatic pathologies. Comparative PET, autoradiographic. and immunohistochemical investigations illustrated the primary contribution of hypertrophic microglia and macrophages to acute TSPO signals in the vicinity of the impact. Astrocytes also formed a TSPO-positive glial scar encompassing necrotic inflammation, and were clustered with PET-detectable TSPO signals in the bilateral external and internal capsules at late stages, putatively reacting with diffuse axonal injury. These observations support the applicability of TSPO-PET as an imaging-based preclinical and clinical biomarker assay in TBI, and indicate its potential capability to clarify aggressive and protective roles of glial responses to injury when combined with emerging anti-inflammatory and immunomodulatory treatments.

Object Recent evidence suggests that a glioma stem cell subpopulation might contribute to radioresistance in malignant gliomas. To investigate this hypothesis, the authors examined recurrent malignant gliomas for histopathological changes after high-dose irradiation with Gamma Knife surgery (GKS) and external beam radiation therapy (EBRT). Methods Thirty-two patients with malignant gliomas (Grade 3 in 8 patients, Grade 4 in 24) underwent GKS in combination with EBRT. Serial MR and L-[methyl-11C] methionine PET images were employed to assess remnant or recurrent tumors after GKS. Twelve patients underwent surgical removal after GKS and EBRT. Histological sections were subjected to immunohistochemistry for MIB-1, factor VIII, and stem cell markers, nestin and CD133. Results The site of GKS treatment failure was local in 16 (76.2%) of 21 patients with glioblastomas showing progression; in 9 of these 16 patients, the recurrence clearly arose within the target lesion of GKS. Histopathological examination after GKS and EBRT showed variable mixtures of viable tumor tissues and necrosis. Viable tumor tissues exhibited high MIB-1 indices but reduced numbers of tumor blood vessels. There was marked accumulation of CD133-positive glioma cells, particularly in remnant tumors within the necrotic areas, in sections obtained after GKS plus EBRT, whereas CD133-positive cells appeared very infrequently in primary sections prior to adjuvant treatment. Conclusions The results indicate that CD133-positive glioma stemlike cells can survive high-dose irradiation, leading to recurrence, despite prolonged damage to tumor blood vessels. This could be an essential factor limiting the effectiveness of GKS plus EBRT for malignant gliomas.

Automatic control systems of brain temperature for water surface-cooling were first-ever applied to the brain hypothermic treatment of patients. A patient in ICU was regarded as a unity controlled system with an input (temperature of water into blanket) and an output (tympanic membrane temperature). The proposed algorithm of optimal-adaptive and fuzzy control laws inclusive of our developed cooling and warming machine were well confirmed during the hypothermic course to keep brain temperature of patients within its allowable range. It was well controlled without much influence due to room temperature, metabolic and circulatory change caused by various medical treatments including the effect of nonlinear and time-varying characteristics of individual patients. The clinical control of brain temperature was almost continuously performed in around 10 days, under the brain temperature between 35 degrees C and 37 degrees C scheduled by physicians according to the state of patients. Their state had been monitored during the therapeutic course of pharmacological treatment with almost everyday examinations by CT imaging, referring various vital signs inclusive of the temperature of urinary bladder with continuous measurement of intracranial pressure by a catheter placement in cerebral sinus. The patients had good recovery to their rehabilitation after mild hypothermia by the proposed automatic control systems.

A 51-year-old man presented with an extremely rare case of intracranial subarachnoid hemorrhage caused by rupture of an anterior spinal artery aneurysm manifesting as disturbance of consciousness following sudden onset of neck pain and numbness of the extremities. Cranial computed tomography revealed subarachnoid hemorrhage, mainly in the posterior fossa. Cerebral angiography studies on admission and on the 4th day demonstrated no definite abnormality as a bleeding source. A ventricular catheter was inserted to treat the acute hydrocephalus, and conservative management was continued during the acute period. Third angiography on the 18th day demonstrated an anterior spinal artery aneurysm at the C1 level which was considered to be the bleeding site. After conservative treatment, the patient was discharged without neurological deficits. Fourth angiography on the 108 th day disclosed spontaneous disappearance of the aneurysm, which was confirmed by the fifth angiography on the 269 th day. If subarachnoid hemorrhage of unknown etiology is encountered, spinal artery aneurysm should be considered as the bleeding source. Despite the controversy concerning the treatment strategy, ruptured spinal artery aneurysms can be treated conservatively because of the possibility of spontaneous regression. Follow-up angiography is required to evaluate the natural course of the lesion.

In vivo cell cycle analysis in higher eukaryotes has been limited by the challenge of preserving the integrity of the living organism while visualizing dividing cells. Here, we propose a new model, which uses the unique combination of features of the Japanese medaka in order to visualize and manipulate the cell cycle progression in a live vertebrate. Our stable transgenic histone H2B-GFP medaka line allows fluorescence-based monitoring of the chromosomes. The system has a high specificity, with a strong GFP signal labeling the chromatin architecture. The subcellular resolution ensures detection of both normal and abnormal divisions in live recordings. This translates into the possibility to quantify temporal and spatial aspects of the cell cycle, such as length or nuclear size, as well as to expose drug toxicity at the earliest stage. We also show that acclimation to cold, a prominent feature of the eurytherm medaka, is a valuable natural way of inducing a reversible cell cycle arrest in the entire living organism. Our results suggest that this manipulation can be performed from the early stages of development, has no toxicity and does not alter the cell cycle profile of the embryo.

Object A multimodal neuronavigation system using metabolic images with PET and anatomical images from MR images is described here for glioma surgery. The efficacy of the multimodal neuronavigation system was evaluated by comparing the results with that of the conventional navigation system, which routinely uses anatomical images from MR and CT imaging as guides. Methods Thirty-three patients with cerebral glioma underwent 36 operations with the aid of either a multimodal or conventional navigation system. All of the patients were preliminarily examined using PET with l-methyl-[11C] methionine (MET) for surgical planning. Seventeen of the operations were performed with the multimodal navigation system by integrating the MET-PET images with anatomical MR images. The other 19 operations were performed using a conventional navigation system based solely on MR imaging. Results The multimodal navigation system proved to be more useful than the conventional navigation system in determining the area to be resected by providing a clearer tumor boundary, especially in cases of recurrent tumor that had lost a normal gyral pattern. The multimodal navigation system was therefore more effective than the conventional navigation system in decreasing the mass of the tumor remnant in the resectable portion. A multivariate regression analysis revealed that the multimodal navigation system–guided surgery benefited patient survival significantly more than the conventional navigation–guided surgery (p = 0.016, odds ratio 0.52 [95% confidence interval 0.29–0.88]). Conclusions The authors' preliminary intrainstitutional comparison between the 2 navigation systems suggested the possible premise of multimodal navigation. The multimodal navigation system using MET-PET fusion imaging is an interesting technique that may prove to be valuable in the future.

We compared the tumor uptake of (11)C-methionine (MET) with positron emission tomography (PET) with the results of a pathological analysis to examine the proliferative potential and microvessel density measured with immunostaining for MIB-1 and factor VIII, respectively, from 33 patients with glioma. The MET uptake in oligodendrogliomas was significantly greater than that in grade 2 astrocytomas and comparable to those in grade 3 and 4 astrocytomas. The MIB-1 index of oligodendroglioma meanwhile was comparable to that of grade 2 astrocytoma. The microvessel area in oligodendroglioma was significantly greater than that in grade 2 astrocytomas and comparable to those in grade 3 and 4 astrocytomas. According to a multivariate statistical analysis, MET uptake ratio was closely correlated with the MIB-1 index among astrocytomas. An increase in the microvessel area in the oligodendrogliomas contributed to the higher MET uptake among the tumors with a low-proliferative index. This information is important for interpreting the results of MET-PET studies for clinical use.

Mammalian nuclear Dbf2-related (NDR) kinases (LATS1 and 2, NDR1 and 2) play a role in cell proliferation, apoptosis and morphological changes. These kinases are regulated by mammalian sterile 20-like kinases (MSTs) and Mps one binder (MOB) 1. Okadaic acid (OA), which activates MST2, facilitates the complex formation of MOB1, MST2 and NDR1 in HEK293FT cells. The in vitro biochemical study demonstrates the phosphorylation of MOB1 by MST2. The phosphorylated MOB1 alone is capable to partially activate NDR1 in vitro, but MST2 is also required for the full activation. The knockdown of MOB1 or MST2 abolishes the OA-induced NDR1 activation in HEK293FT cells. Among MOB1 mutants, in which each serine or threonine residue is replaced with alanine, MOB1 T74A and T181A mutants fail to activate NDR1. Thr74, but not Thr181, is phosphorylated by MST2 in vitro, although MOB1 is also phosphorylated by MST2 at other site(s). The interaction of MOB1 T74A with NDR1 is barely enhanced by OA treatment. These findings indicate that the phosphorylation of MOB1 at Thr74 by MST2 is essential to make a complex of MOB1, MST2 and NDR1, and to fully activate NDR1.

We designed the present experiments to investigate the involvement of endogenous nitric oxide synthase (NOS) inhibitors, dimethylarginine dimethylaminohydrolase (DDAH) as a hydrolyzing enzyme of the NOS inhibitors, NOS, arginase which shares l-arginine as a common substrate with NOS, and endothelin-1 (ET-1) in the pulmonary dysfunction after induction of experimental subarachnoid hemorrhage (SAH) in the rabbit. SAH was induced by injecting autologous blood into the cisterna magna, and controls were injected with saline. On day 2, pulmonary arteries were isolated for determinations. A significant impairment of the endothelium-dependent relaxation (EDR) caused by acetylcholine was found in 20 cases (43.5%) out of 46 SAH animals, and the same animals exhibited accompanying the significantly impaired cyclic GMP production, accumulated endogenous NOS inhibitors, attenuated DDAH activity, enhanced arginase activity and accumulated ET-1 within the vessel wall. Meanwhile, there were no differences in endothelial NOS activity per se and sodium nitroprusside-induced relaxation between the animals with an impaired EDR and those without such a change. ET-1 content within aortic wall was increased with concomitant decrease in cyclic GMP production after the intraperitoneal application of authentic monomethylarginine as a NOS inhibitor in the rat. The current results suggest that accumulated endogenous NOS inhibitors and enhanced arginase activity possibly bring about the impaired NO production, thereby attenuating the EDR and contributing to the accumulation of ET-1 within the vessel wall. The accumulated endogenous NOS inhibitors at least partly result from the decreased DDAH activity. These alterations may be relevant to the pulmonary dysfunction after induction of SAH.

The management of clival chordoma remains problematic. We present the case of a 48-year-old woman with clival chordoma who underwent multiple surgeries and radiation therapy, including gamma knife stereotactic radiosurgery (GK-SRS), during a 10-year clinical course. The tumor was initially removed by gross total resection via the trans-sphenoidal approach, followed by external linac radiation therapy. The tumor recurred at the clivus 5 years after the initial operation. After repeated trans-sphenoidal removal of recurrent tumors, she twice underwent GK-SRS for a tumor remnant adjacent to the brainstem. Although this part of the tumor was controlled by GK-SRS, there was further tumor extension toward the sphenoid and maxillary sinuses. Ultimately, lower cranial nerve dysfunction developed due to tumor extension into the lower part of the clivus and the patient died of respiratory failure. Autopsy revealed the tumor to extend from the lower clivus to the bilateral middle fossae. The lower part of the tumor extended to the nasal cavity and to the posterior wall of the pharynx, resulting in compression of the upper pharyngeal region. The tumor around the jugular foramen compressed the lower cranial nerves bilaterally. Tumor cells did not, however, invade the intradural space microscopically. Although chordoma is not biologically malignant, this tumor can show massive extension with destruction of bony structures and extracranial invasion of connective tissues. Therefore, the optimal treatment strategy is to remove the tumor mass as extensively as possible, including normal bony structures and connective tissues surrounding the tumor, using skull base surgical techniques.

The present study was designed to investigate whether endothelial intracellular calcium concentration ([Ca(2+)](i)), endothelial nitric oxide synthase (eNOS) activity and nitric oxide (NO) generation altered in association with impaired endothelium-dependent relaxation (EDR) in pulmonary artery (PA) specimens from experimental subarachnoid hemorrhage (SAH) rabbits. Injecting non-heparinized autologous arterial blood into cisterna magna induced the SAH. Simultaneous measurements of endothelial [Ca(2+)](i) and isometric tension of PA specimens were performed using fura 2. The subjects included normal control rabbits (group N), SAH rabbits with normal EDR (group A) and with impaired EDR (group B). When treated with 10(-7) M acetylcholine (ACh), endothelial [Ca(2+)](i) was significantly lower in group B (74.1+/-8.5 nM) than that in groups A (153.0+/-28.0 nM, p0.05) and N (184.8+/-27.8 nM, p0.01). Basal and ACh-stimulated cyclic GMP productions as a marker of NO generation were also significantly (p0.005) decreased in group B as compared to those in the other two groups. Meanwhile, there were no differences in eNOS activity per se among the three groups. These results suggest that the attenuated endothelial [Ca(2+)](i) elevation leads to the impaired NO generation in PA endothelium, which in turn impairs the EDR and possibly increases the vascular resistance of PA following SAH.

✓Magnetic resonance (MR) imaging is a powerful tool for detecting and characterizing ischemic edema, a serious complication of ischemic cerebrovascular disorders. In this article the authors investigate the relationships between MR imaging findings and structural/ultrastructural changes in ischemic brain edema by using various animal models of experimental cerebral ischemia. The authors observed the following: 1) Ischemic edema was detectable by diffusion weighted MR imaging as early as 15 minutes after the onset of vascular occlusion. A decrease in the apparent diffusion coefficient (ADC) corresponded to the early cellular/cytotoxic type of brain edema and the decrease was proportionate to the degree of intracellular water accumulation. 2) Postischemic transient normalization of the ADC after the initial decrease did not signify histological recovery but rather slowly progressing infarction. 3) Histological degradation of postischemic tissue correlated with the decrease in tissue elasticity and the magnetization transfer ratio. 4) Transient cytotoxic edema localized in the substantia nigra preceding neuronal death was detectable on MR images after ipsilateral striatal infarction. Thus, MR imaging is a powerful tool for detecting and characterizing brain edema associated with ischemic stroke.

The object of this study was to analyze the pathology of collateral vessels newly induced by indirect bypass surgery for moyamoya disease (MMD). An autopsy analysis was conducted on a 39-year-old woman with MMD who had died of a brainstem infarction. The patient had undergone bilateral indirect bypass surgeries 22 years earlier. Sufficient revascularization via bilateral external carotid arterial systems was confirmed by cerebral angiography before her death. Macroscopic observation of the operative areas revealed countless meandering vessels on the internal surface of the dura mater connected with small vessels on the brain surface and in the subpial brain tissue. Notably, microscopic analysis of these vessels revealed the characteristic 3-layer structure of an arterial wall. This autopsy analysis was the first to confirm that indirect bypass surgery had induced the formation of a new arterial network (arteriogenesis) and that this network had been maintained for more than 20 years to compensate for the chronic cerebral ischemia caused by the MMD.

OBJECT The majority of sinonasal malignancies present with advanced disease, and cure rates are generally poor. Surgical extirpation remains the mainstay of treatment. In cases of sinonasal malignancy with orbital apex extension, gross-total tumor resection requires orbital exenteration and bony skull base resection around the orbital apex to provide sufficient margins. In this retrospective study, the authors describe their surgical strategy in and technique for orbital exenteration with orbital apex resection in patients at Tokyo Medical and Dental University who had sinonasal malignancy with orbital apex extension. They also analyzed the clinical features of and the results in these patients. METHODS Between February 2001 and August 2012 at the authors' institution, sinonasal malignancy with orbital apex extension was treated using craniofacial tumor resection with orbital exenteration including skull base bone around the orbital apex. The authors describe this technique and analyze the surgical indications, extent of resection, primary tumor location, outcome, pathological findings, and neoadjuvant and adjuvant therapies of the patients who underwent the technique. RESULTS The patients consisted of 12 men and 3 women with a mean age of 47.7 years (range 14–79 years). The longest postoperative follow-up was 9.5 years, and the shortest was 0.67 year (mean 3.0 years). Tumor originated at the ethmoid sinus in 6 patients (40%), maxillary sinus in 5 (33%), nasal cavity in 2 (13%), and orbital cavity and maxillary bone in 1 patient each (7%). Histological analysis of tumor specimens revealed squamous cell carcinoma in 9 patients (60%), rhabdomyosarcoma in 2 (13%), and small cell carcinoma, mucoepidermoid carcinoma, adenoid cystic carcinoma, and Ewing sarcoma in 1 patient each (7%). Two patients experienced recurrences at 1 and 5 months after treatment; these patients died at 5 and 10 months after surgery, respectively. Estimated 5-year recurrence-free survival (RFS) was 86.7%, and estimated 5-year overall survival (OS) was 86.2%; there was no perioperative mortality. None of the patients had new neurological deficits as a result of the surgery, but 5 patients suffered infectious complications from the graft transplanted into the cavity after resection. There were no other perioperative complications. CONCLUSIONS These authors are the first to describe a technique for extended orbital exenteration with orbital apex skull base resection. The technique provided sufficient margins for gross-total resection of the sinonasal malignancy with orbital apex extension. The estimated 5-year OS and RFS rates were high, and the perioperative complication rate was acceptably low, demonstrating the safety and efficacy of this technique.

Posttraumatic hyperactivity is a neurobehavioral symptom commonly seen in patients after traumatic brain injury (TBI). No useful animal model has yet been established for evaluation of this important symptom. We induced either mild (MILD, 0.7-0.9 atm) or moderate (MOD, 1.3-1.6 atm) lateral fluid percussion injury (LFPI) in Mongolian gerbils. Open-field and T-maze tests were used during a 7-day period after the trauma. All animals were perfusion fixed for histopathological examinations. Transient locomotor hyperactivity was found with a peak at 6 hr after injury in the MILD animals, whereas MOD animals showed prolonged and severe hyperlocomotion throughout the 7-day posttrauma period (P < 0.0001). Interestingly, the temporal profile of the posttraumatic hyperactivity was similar to that of the working memory deficit in both injury groups. Histological examination revealed significant neural tissue damages, including cortical necrosis, white matter rarefaction, and neuronal loss in the hippocampus in the ipsilateral hemisphere of the MOD animals, vs. only negligible changes in the MILD animals. Correlation analysis revealed that the volume of white matter lesions was significantly correlated with both posttraumatic hyperactivity (r = 0.591, P < 0.01) and working memory deficit (r = -0.859, P < 0.0001). Taken together, our findings confirm the successful reproduction of posttraumatic hyperactivity following experimental TBI. The posttraumatic hyperlocomotion probably shared pathomechanisms common to those of cognitive dysfunction caused by LFPI, supporting the speculation from previous studies that some neurobehavioral abnormities intimately correlate with TBI-induced cognitive dysfunction. Histopathologically, significant involvement of white matter damage in the posttraumatic functional deficits was indicated.

We evaluated correlation between neurochemical and functional alterations of the nigrostriatal dopaminergic system in rat brains lesioned with 6-hydroxydopamine (6-OHDA), that model hemi-Parkinson's disease (PD), by using three different quantitative in vivo and in vitro methods. Rats unilaterally lesioned with different doses of 6-OHDA underwent two types of in vivo experiments: (1) a rotational behavioral study with methamphetamine (MAP) or apomorphine (APO); and (2) a positron emission tomography (PET) study with [11C]PE2I (radioligand for dopamine transporters) or [11C]raclopride (radioligand for dopamine D2 receptors). An in vitro autoradiographic study with the same radioligands was also conducted. The number of rotations after the MAP or APO injection increased with increased doses of 6-OHDA. The in vitro and in vivo binding of [11C]PE2I dose-dependently decreased in response to the 6-OHDA injections, while that of [11C]raclopride dose-dependently increased. There was a significant negative hyperbolic correlation between the number of rotations after MAP injection and the binding of [11C]PE2I. In contrast, there was a significant positive linear correlation between the number of rotations after APO injections and the binding of [11C]raclopride. These results robustly reveal a molecular pharmacological basis of parkinsonian symptoms in animal models of PD, and indicate the utility and validity of in vivo PET measurements in assessing pre- and post-synaptic dopaminergic functions.

The authors retrospectively analyzed the data obtained in patients who had undergone L-[methyl-11C] methionine (MET)-positron emission tomography (PET) studies to clarify the relationship between MET uptake and tumor biological features and to discuss the clinical usefulness of MET-PET studies.One hundred ninety-four patients with cerebral glioma or suspected glioma underwent PET scanning 20 minutes after injection of MET, whose uptake into the tumor was expressed as a ratio to contralateral healthy brain tissue (T/N ratio). Analyses were performed to determine how MET uptake correlated with tumor pathological features and prognosis. The T/N ratios before and after various treatments were also examined. There were significant differences in the T/N ratio among the nonneoplastic lesions, low-grade gliomas, and malignant gliomas. Furthermore, there were significant correlations between patient survival and pretreatment T/N ratios. Among patients with malignant gliomas, a significant difference in survival was observed between cases with and without postoperative tumor remnant based on elevated MET uptake. The MET uptake was heterogeneous even among the homogeneous tumor areas demonstrated on MR imaging. Malignant pathological features were detected in the areas with the highest MET uptake. The effectiveness of radiotherapy or chemotherapy was expressed as a significantly decreased T/N ratio in some of the tumor types.The ability of MET-PET to reflect the biological nature of gliomas makes it an excellent method for monitoring active tumor tissue, and treatments based on its findings should provide a powerful clinical protocol in the course of glioma therapy.

We examined whether the degree of hemodynamic stress in patients with chronic occlusive cerebral vascular disease can be quantitatively evaluated with the use of perfusion-weighted magnetic resonance imaging (PWI). Thirty-six patients with moyamoya disease (mean age, 26.8 years; range, 18 to 59) underwent PWI and positron emission tomography (PET) within a month's interval. The PWI data were calculated by three different analytic methods. The cerebral blood flow (CBF) ratio, cerebral blood volume (CBV) ratio, and mean transit time (MTT) of the anterior circulation were calculated using the cerebellum as a control region and compared with PET data on the same three parameters and oxygen extraction fraction (OEF). Parametric maps of PWI attained a higher resolution than the PET maps and revealed focal perfusion failure on a gyrus-by-gyrus level. The relative CBV and MTT obtained with PWI showed significant linear correlations with the corresponding PET values (CBV, R2 = 0.47 to 0.58; MTT, R2 = 0.32 to 0.68). We also found that we could detect regions with abnormally elevated OEF and CBV based on the delay of PWI-measured MTT relative to the control region by defining a 2.0-sec delay as a threshold. The sensitivity and specificity were 92.3% and 100% in detecting regions with abnormally elevated OEF, and 20.0% and 100% in detecting regions with abnormally elevated CBV, respectively. Among the parameters obtained with PWI, our results suggested that the relative CBV value and delay of MTT might be quantitatively manipulated to assist in clinical decision-making for patients with moyamoya disease.

Background: The optimum management of patients with moyamoya disease remains controversial. Objectives: To examine retrospectively the correlation between the degree of haemodynamic stress and the clinical presentation by measuring cerebral haemodynamics and metabolism using positron emission tomography (PET). Methods: 57 patients with moyamoya disease (mean age 32 years, range 12 to 64), classified into five groups according to clinical manifestations, underwent PET measurement of cerebral blood flow (CBF), cerebral blood volume (CBV), cerebral metabolic rate for oxygen (CMRO2), and oxygen extraction fraction (OEF) using 15O labelled gases. The regional values in patient groups were compared with a normal group. Results: CBF in non-symptomatic patients, patients presenting with transient ischaemic attacks (TIA), and patients with haemorrhagic onset (H) was not significantly lower than in normal controls in any region. CBV in the TIA group and in patients with infarction associated with TIA (I/TIA) was significantly higher than in the controls in most regions. OEF in the frontal, parietal, and temporal cortex was significantly higher in the I/TIA group than in the controls. Patients in the H group and those with a permanent deficit with infarction (PD group) had decreased metabolism with normal OEF. Multivariate analysis to test the distribution of the three dimensional vector (CBF, CBV, OEF) showed significant differences between every possible pair among the six groups except NS v H and H v PD in the frontal cortex. Conclusions: The haemodynamic status of moyamoya disease is not uniform, and severe haemodynamic stress occurs in selected subgroups of patients.