Microendophenotypes of psychiatric disorders: phenotypes of psychiatric disorders at the level of molecular dynamics, synapses, neurons, and neural circuits.

Psychiatric disorders are caused not only by genetic factors but also by complicated factors such as environmental ones. Moreover, environmental factors are rarely quantitated as biological and biochemical indicators, making it extremely difficult to understand the pathological conditions of psychiatric disorders as well as their underlying pathogenic mechanisms. Additionally, we have actually no other option but to perform biological studies on postmortem human brains that display features of psychiatric disorders, thereby resulting in a lack of experimental materials to characterize the basic biology of these disorders. From these backgrounds, animal, tissue, or cell models that can be used in basic research are indispensable to understand biologically the pathogenic mechanisms of psychiatric disorders. In this review, we discuss the importance of microendophenotypes of psychiatric disorders, i.e., phenotypes at the level of molecular dynamics, neurons, synapses, and neural circuits, as targets of basic research on these disorders.

SU-E-J-33: Geometric Agreement Check for Imaging System with Radiation Beam by KV and MV-CBCT.

PURPOSE: The verification method of the geometry agreement between a light field and/or a laser coordinate and treatment beam should be easy and quick. In this presentation, we propose a novel QA method by using both kV- and MV-CBCT for kV-IGRT system. This method confirms the temporal unchanging the agreement of geometry in the kV-IGRT system with the treatment beam geometry. METHODS: 1) MV-flexmap: Sequential MV-projection images were acquired during gantry rotation by iViewGT (Elekta) and MV-CBCT was reconstructed by in-house software with a flexmap correction. The flexmap is displacement of gantry and detector panel related with gantry sag. The geometric change affects the deranging reconstructed image. To evaluate how much displacement of EPID panel and gantry was detectable, the images of 8mm diameter ball-bearing (BB) located at the radiation isocenter were reconstructed with improper Flexmap.2) A comparison between the kV-CBCT and the MV-CBCT: The kV-CBCT was provided by X-ray Volume image (XVI) system (Elekta). To confirm the agreement for the geometry between kV-IGRT system and treatment beam, the kV-CBCTs of BB are compared with that of MV-CBCTs. RESULTS: The flexmaps were modified to (b)1mm / (c)3mm shifted to the rotation direction and (d)3mm to the rotation axis. The MV-CBCT were reconstructed with the correct flexmap and with incorrect flexmap (b), (c) and (d).　The geometric confirmation for MV-CBCT was done by comparison of the width and center of the BB on the MV-CBCT. The discrepancy of center between kV-CBCT and MV-CBCT was less than 1mm. CONCLUSIONS: Less than 1mm of the geometrical changing to rotation direction for MV-detector panel could be recognized by reconstructed images of BB. Using kV- and MV-CBCT enable us to perform the simple comparison for geometrical non-idealities between the kV-IGRT system and the treatment beam. Dr. K. Nakagawa received research grant from Elekta.

SU-E-J-203: Determination of PTV Margin for Lung Tumor Using In-Treatment 4D CBCT.

PURPOSE: To determine a planning target volume (PTV) margin for lung cancer patients using a four-dimensional cone-beam CT (4D CBCT) acquired during volumetric modulated arc therapy (VMAT) treatment. METHODS: A VMAT plan for lung cancer patients was created by Pinnacle v9.0 (Philips) treatment planning system (TPS), where the gross target volume (GTVs) in each breathing phase was delineated by using 4D-planning CT scan (TOSHIBA and ANZAI). The VMAT treatment was performed with a stereotactic body frame after the registration using Elekta X-ray volume imaging (XVI) unit. Simultaneous cone-beam projection images were acquired for 3 or 4 fractions of 10 patients. The in-treatment 4D CBCT was reconstructed by dividing into four breathing phase bins. A total of 38 in-treatment 4D-CBCT sets were exported to Pinnacle TPS. The isocenter of in-treatment 4D CBCT was matched with that of 4D-planning CT. The tumor motion during treatment was manually tracked on in-treatment 4D CBCT, and the center-of-mass (COM) location of the tumor was estimated. Analyzing the tumor regions observed by in-treatment 4D CBCT, a PTV margin in our system was derived. RESULTS: The average difference in COM location of the tumor was less than 1mm for all directions, while the standard deviations (SD's) were about 1.3mm, 1.6mm, and 2.1mm for the lateral, the vertical, and the longitudinal directions, respectively. The large discrepancy more than 3mm was observed for one patient. The required PTV margin was about 3-4mm for the lateral and the vertical directions, whereas it was about 5mm for the longitudinal direction. CONCLUSIONS: The uncertainties of the tumor motion caused by respiration were observed by in-treatment 4D CBCT images. It was feasible to determine the PTV margin from 4D volume images. K. Nakagawa receives research funding from Elekta.

Tight regulation of transgene expression by tetracycline-dependent activator and repressor in brain.

Methods to temporally and spatially regulate gene mutations will provide a powerful strategy to investigate gene function in the brain. To develop these methods, we have established a tightly regulated system for transgene expression in the forebrain using both a tetracycline (Tc)-dependent transcription activator (rtTA) and a repressor (TetR-Kruppel-associated box). In this system, the repressor binds to the Tc-responsive element (TRE) in the absence of doxycycline (Dox), leading to the repression of leaky activation of TRE-mediated transcription caused by weak binding of rtTA to TRE. Upon Dox administration, only the activator binds to TRE and activates transcription. We tested this system in cultured cells by bicistronically expressing both the regulators using an internal ribosome entry site (IRES). In COS-1, HeLa and SHSY5Y cells, leaky transcription activation led by rtTA in the absence of Dox was repressed without decreasing the level of activated transcription in the presence of Dox. Using this system, transgenic mice were produced that express both the regulators using IRES in the forebrain under the control of the alphaCaMKII promoter and were bred with transgenic mice carrying the TRE-dependent reporter transgene. In reverse transcription-polymerase chain reaction and in situ hybridization analyses of the forebrain in adult double transgenic mice, the treatment of Dox induces reporter mRNA expression, which was not detected before the treatment and after the withdraw of Dox following the treatment. These results indicate that this system allows the tight regulation of transgene expression in a Dox-dependent fashion in the forebrain and will be useful in investigating gene function in the brain.

Surgical pathology of spinal schwannoma: has the nerve of its origin been preserved or already degenerated during tumor growth?

OBJECTIVE: This study was aimed to understand ultrastructural pathology of nerves of tumor origin of spinal schwannomas, which has not been reported so far, in order to understand the mechanism of the postoperative functional restoration after the nerve transection. METHODS: From 13 patients who underwent sacrifice of an affected nerve root at total removal of spinal schwannomas (C2 conus), the proximal (spinal cord side, n = 12) and distal (dorsal root ganglion side, n = 10) stumps of the nerves of the tumor origin were collected and examined by light and electron microscope, followed by morphometric analysis (n = 9). RESULTS: Almost all of affected nerves at both proximal and distal to the lesion were composed of well-preserved myelin sheath and axons with mild disturbance of endo- and perineurial structures at light microscopic level except one case, which showed severe fibrosis. Electron-microscopically, regenerated axons with thin myelin were found in part in the proximal and distal nerves with few macrophages in three cases. The area of nerves (mm2), density of myelinated axons (axons/mm2) and total number of myelinated axons in the proximal stump (0.552 +/- 0.430, 10,400 +/- 5,240 and 5,480 +/- 4,790) was approximately 70%, 80% and 60%, respectively, of those in the distal stump (0.765 +/- 0.333, 12,400 +/- 5,180 and 9,970 +/- 8,630). CONCLUSIONS: This data combined with no permanent deficits after nerve transection suggest that the nerves of tumor origin are in the processes of slowly progressed deterioration with repeated degeneration and regeneration/remyelination, and the postoperative rapid recovery from the transient neurological deficit may be explained by functional compensation by the adjacent non-affected nerves with slow tumor growth.

Inducible repression of CREB function disrupts amygdala-dependent memory.

Evidence from Aplysia, Drosophila, mice, and rats indicates that the CREB (cAMP/Ca2+ responsive element binding protein) family of transcription factors is critical for long-term memory. Recent findings, however, suggest that performance abnormalities may contribute to the memory deficits attributed to CREB manipulations in mammals. To clarify the role of CREB in memory, we used a paradigm, conditioned taste avoidance, that places few performance demands on the subject. We show that lesioning or blocking protein synthesis in the basolateral amygdala of mice disrupts conditioned taste aversion. Furthermore, either chronically or acutely disrupting CREB function in two different types of genetically modified mice blocks memory for conditioned taste aversion measured 24 h following training. Together, these findings indicate that CREB-mediated transcription and protein synthesis are required for conditioned taste aversion memory.

Contrast-enhanced MRA for investigation of cerebral arteriovenous malformations.

We evaluated contrast-enhanced MRA (enhanced 3-D fast gradient-echo [efgre3d] with spectral inversion recovery) for identification of 15 intracranial arteriovenous malformations (AVMs) in 14 patients. Demonstration of the feeding arteries was classified as good for 16 examinations on maximum-intensity projections and multiprojection volume reconstruction images. The nidus was seen well in all patients. Definition of the draining veins was good or fair except for one poor result. Therapeutic effects were clearly demonstrated in three follow-up series. Contrast-enhanced MRA using efgre3d is useful for delineation of AVMs and for follow-up after treatment.

Expression of E-cadherin-catenin complex in human benign schwannomas.

The Ca2+-dependent cell adhesion molecule E-cadherin has been known to express in normal and reactive Schwann cells in rodents, and to play an important role in Schwann cell-Schwann cell adhesion and maintenance of peripheral nervous tissue architecture. However, little is known about expression of E-cadherin in schwannomas. The aim of the present study was to investigate the cellular expression and localization of E-cadherin, and its associated protein, alpha E-, alpha N- and beta-catenins in human schwannomas, which are supposed to derive from Schwann cells. We tested the hypothesis that these proteins might show an altered expression/distribution in schwannoma cells which correlates with their neoplastic behavior, including sparse cell-cell contact, as seen those in meningiomas and various carcinomas. In human schwannomas, however, E-cadherin, alpha E-catenin, and beta-catenin were detected by western blotting and immunohistochemistry, whereas alpha N-catenin was not. Immunoprecipitation using anti-E-cadherin antibody resulted in alpha E-catenin forming a complex with E-cadherin. SSCP analysis revealed no mutations in the transmembrane domain or in intracellular catenin-binding site of E-cadherin. These data suggest that the E-cadherin-alpha E-catenin complex is well preserved in human schwannoma cells, which is compatible with its benign behavior, and these molecules might be used as additional cell markers of Schwann cell-derived tumors.

CT during selective arteriography: anatomical assessment of unruptured intracranial aneurysms before endovascular treatment.

Our aim was to investigate the usefulness of helical CT during selective angiography (CT arteriography) in pretreatment assessment of unruptured intracranial aneurysms. We studied 47 unruptured aneurysms in 34 prospectively recruited patients for whom endovascular embolisation was initially considered. As pretreatment assessment, we performed rotational digital subtraction angiography (DSA) followed by CT arteriography. The findings on axial source images (axial images) and reconstructed three-dimensional CT angiography (3D-CTA) of CT arteriography were compared to those of rotational DSA, with particular attention to the neck of the aneurysm and arterial branches adjacent to it. Information provided by CT arteriography was more useful than that of rotational DSA as regards the neck in 25 (53%) of 47 cases and as regards branches in 18 (49%) of 37 aneurysms. On axial images, small arteries such as the anterior choroidal artery were seen in some cases. CT arteriography can provide valuable additional information about unruptured aneurysms, which cannot be obtained by rotational DSA alone. This technique is useful for obtaining anatomical information about aneurysm anatomy and for deciding the therapeutic strategy.

Tocopherol-associated protein is a ligand-dependent transcriptional activator.

Vitamin E is a term that encompasses a group of potent, lipid-soluble, chain-breaking antioxidants. Structural analysis reveals that molecules having vitamin E activity include four isomers (alpha, beta, gamma, and delta) of both tocopherols and tocotrienols. Alpha-tocopherol has been shown to have the highest biological vitamin E activity in mammalian tissues based on fetal resorption assays, and it reverses vitamin E deficiency symptoms. Although the molecular functions fulfilled specifically by alpha-tocopherol have yet to be fully described, it is unlikely that they are limited to general antioxidant functions. Here we show the functional characterization of alpha-tocopherol associated protein, TAP, which displays significant sequence similarity to the alpha-tocopherol transfer protein. Ligand competition analysis showed that recombinant TAP binds to alpha-tocopherol but not to other isomers of tocopherols. Using GFP fusion protein expression system, we observed that TAP translocates from cytosol to nuclei in alpha-tocopherol-dependent fashion. Transient transfection experiment showed that TAP activates transcription of the reporter gene in alpha-tocopherol-dependent manner. These results suggest that the biological function of alpha-tocopherol is not only as an antioxidant but also as a transcriptional regulator of gene expression via association with a transcription factor TAP.

Improved cerebral perfusion and metabolism after stenting for basilar artery stenosis: technical case report.

OBJECTIVE: Recent advances in stent technology have allowed the introduction of more flexible stents that may be tracked more easily in the intracranial vessels. We present a patient with improved cerebral blood flow and metabolism as assessed by positron emission tomography after stent-assisted angioplasty for symptomatic basilar artery stenosis. CLINICAL PRESENTATION: A 62-year-old man, who had undergone left superficial temporal artery to middle cerebral artery bypass surgery for left internal carotid artery occlusion 10 years previously, presented with dizziness, blurred vision, and memory disturbance. Angiography revealed severe stenosis of the proximal basilar artery. Positron emission tomographic scans revealed decreased cerebral blood flow associated with increased oxygen extraction fraction in the entire brain, particularly in the posterior circulation and the left middle cerebral artery territory. Despite medical treatment, the patient experienced worsening visual disturbance and right-sided motor weakness. INTERVENTION: Ticlopidine and aspirin were used as antiplatelet agents. In addition, we used argatroban, which is a direct thrombin inhibitor, as an anticoagulant during the procedure. Predilation with a coronary artery balloon was performed, followed by placement of a GFX 3- by 8-mm stent (Arterial Vascular Engineering, Santa Rosa, CA), with excellent angiographic results. The patient made a good neurological recovery, and the postoperative positron emission tomographic scan demonstrated increases in both cerebral blood flow and cerebral metabolic rate of oxygen with a normalization of oxygen extraction fraction. CONCLUSION: Stent-assisted angioplasty can provide a favorable clinical course as well as improved cerebral perfusion and metabolism for a patient with basilar artery stenosis. Long-term follow-up data and additional clinical experience are required to assess the durability of this approach.

Surgical pathology of spinal schwannomas: a light and electron microscopic analysis of tumor capsules.

OBJECTIVE: Although spinal schwannomas are often encountered, the pathology of the tumor capsule has not been reported. In this study, we describe the ultrastructural pathology of the tumor capsule of spinal schwannomas. METHODS: In 18 patients who underwent total removal of spinal schwannomas (C2-conus), the tumor capsule was collected and examined by light and electron microscopy. RESULTS: The thickness of the tumor capsule ranged from 15 to 800 microm (mostly 30-100 microm) and was composed of three layers from the surface to the center: 1) a thinly stretched nerve tissue layer; 2) a fibrous layer of fibrocytes, abundant collagen fibers, and tumor vessels; and 3) a thin transitional layer intermingled with fibrous components and tumor cells. The thickness of each layer varied in different regions of the surface. There was no clear separation between the tumor capsule and the neoplastic tissue, even on the electron microscopic level. A number of nerve fibers ran through the fibrous layer and beneath the capsule as well as in the nerve tissue layer. CONCLUSION: Compared with vestibular schwannomas, which have been reported to be covered by an extremely thin layer (3-5 microm) of connective tissue, spinal schwannomas were well encapsulated. The capsule was composed of three distinct components; however, the cleavage between thin capsule and tumor cells was indistinct, and the thickness of the axon-containing capsule varied from site to site. Therefore, resection of the nerve of tumor origin, rather than enucleation, would be justified to avoid tumor recurrence. Surgeons should be aware of this pathology when performing the procedure.

Ruptured irregularly shaped aneurysms: pseudoaneurysm formation in a thrombus located at the rupture site.

OBJECT: The authors describe the clinical, radiological, and pathological findings of ruptured cerebral aneurysms with irregular configurations. METHODS: Eight patients with subarachnoid hemorrhage due to ruptured irregularly shaped aneurysms were examined. The preoperative radiological findings in these cases were compared with the pathological and operative findings of endovascular or open surgery. All of the aneurysms exhibited delayed opacification and delayed washout of contrast medium from the irregularly shaped portion of the aneurysm on digital subtraction angiography and/or helical computerized tomography scanning. Endovascular embolization with platinum coils was attempted in the first four patients who underwent treatment. In three of these patients the aneurysm ruptured again during the endovascular procedure. In the fourth patient an intraaneurysm thrombus was observed during the procedure and clipping was performed. In the subsequent four patients, three underwent clipping without complication and one underwent partial aneurysm embolization because of poor general status. A thrombus adjacent to the aneurysm dome was observed in the patients who underwent open surgery. Pathological examination of the operative specimens revealed a pseudoaneurysm-like cavity in the thrombus that was adherent to the aneurysm. CONCLUSIONS: Ruptured irregularly shaped aneurysms may be accompanied by fragile pseudoaneurysm-like cavities located at the rupture point. Because these aneurysms have a high risk of repeated rupture during an endovascular procedure, advancing microinstruments to the weaker portion of the aneurysm should be avoided.

Cerebellar hemorrhage after coil embolization for a ruptured vertebral dissecting aneurysm.

BACKGROUND: We present a case of ruptured vertebral dissecting aneurysm that exhibited cerebellar hemorrhage after successful embolization of the vertebral artery including the dissected site. CASE PRESENTATION: A 59-year-old man suffered a sudden onset of severe occipital headache when he looked up. Computed tomography demonstrated subarachnoid hemorrhage. Angiography revealed a right vertebral dissecting aneurysm distal to the posterior inferior cerebellar artery. Endovascular embolization of the aneurysm was performed with preservation of the posterior inferior cerebellar artery. The next day, the patient suffered a cerebellar hemorrhage in the vermis. The intracranial pressure was controlled by external ventricular drainage. The patient was discharged with mild cerebellar ataxia and bilateral abducens nerve palsy. CONCLUSION: In a case of vertebral dissecting aneurysm distal to the posterior inferior cerebellar artery, blood circulation in the vertebral arterial system may change after embolization of the aneurysm. In our case, the preserved posterior inferior cerebellar artery might have been hemodynamically stressed postoperatively, resulting in cerebellar hemorrhage. Therefore, strict control of blood pressure is essential in the acute stage after occlusion of the aneurysm.