Genetic study of intracranial aneurysms.

BACKGROUND AND PURPOSE: Rupture of intracranial aneurysms (IAs) causes subarachnoid hemorrhage, leading to immediate death or severe disability. Identification of the genetic factors involved is critical for disease prevention and treatment. We aimed to identify the susceptibility genes for IAs. METHODS: Exome sequencing was performed in 12 families with histories of multiple cases of IA (number of cases per family ≥3), with a total of 42 cases. Various filtering strategies were used to select the candidate variants. Replicate association studies of several candidate variants were performed in probands of 24 additional IA families and 426 sporadic IA cases. Functional analysis for the mutations was conducted. RESULTS: After sequencing and filtering, 78 variants were selected for the following reasons: allele frequencies of variants in 42 patients was significantly (P<0.05) larger than expected; variants were completely shared by all patients with IA within ≥1 family; variants predicted damage to the structure or function of the protein by PolyPhen-2 (Polymorphism Phenotyping V2) and SIFT (Sorting Intolerance From Tolerant). We selected 10 variants from 9 genes (GPR63, ADAMST15, MLL2, IL10RA, PAFAH2, THBD, IL11RA, FILIP1L, and ZNF222) to form 78 candidate variants by considering commonness in families, known disease genes, or ontology association with angiogenesis. Replicate association studies revealed that only p.E133Q in ADAMTS15 was aggregated in the familial IA cases (odds ratio, 5.96; 95% confidence interval, 2.40-14.82; P=0.0001; significant after the Bonferroni correction [P=0.05/78=0.0006]). Silencing ADAMTS15 and overexpression of ADAMTS15 p.E133Q accelerated endothelial cell migration, suggesting that ADAMTS15 may have antiangiogenic activity. CONCLUSIONS: ADAMTS15 is a candidate gene for IAs.

Simultaneous administration of recombinant tissue plasminogen activator and edaravone in acute cerebral ischemic stroke patients.

Among the 1052 patients admitted to our hospital because of cerebral infarction between January 1, 2007, and December 31, 2010, we report the treatment outcomes of 48 patients (4.6% of all patients) who received recombinant tissue plasminogen activator (rt-PA) therapy (simultaneously combined with edaravone) within 3 hours after the onset of infarction. Twenty (41.7%) patients started receiving edaravone before rt-PA administration, and 28 patients (58.3%) started receiving rt-PA and edaravone simultaneously. The patients had an average age of 73.5 years (range, 55-93 years; male:female, 32:16). Medical histories included hypertension, diabetes mellitus, dyslipidemia, arterial fibrillation, and a smoking history in 23 (47.8%), 7 (14.6%), 8 (16.7%), 29 (60.4%), and 8 (16.7%) of patients, respectively. Regarding the treatment outcome of the therapy, the National Institutes of Health Stroke Scale score, which was 15 points before rt-PA administration, showed a statistically significant improvement to 8 points after rt-PA administration (P < .001). The modified Rankin Scale scores at 90 days after treatment were as follows: 0 in 12 patients (25.0%), 1 in 11 patients (22.9%), 2 in 7 patients (14.6%), 3 in 5 patients (10.4%), 4 in 6 patients (12.5%), 5 in 5 patients (10.4%), and 6 in 2 patients (4.2%). The occluded blood vessel reopened completely in 30 patients (62.5%) and partially in 5 patients (10.4%). Asymptomatic hemorrhage over the entire brain developed in 2 patients (4.2%). Thus, rt-PA therapy in combination with edaravone improved the recanalization rate, reduced the incidence of intracranial hemorrhage, and improved functional prognosis.

Pathogenic diversity of Phytophthora sojae and breeding strategies to develop Phytophthora-resistant soybeans.

Phytophthora stem and root rot, caused by Phytophthora sojae, is one of the most destructive diseases of soybean [Glycine max (L.) Merr.], and the incidence of this disease has been increasing in several soybean-producing areas around the world. This presents serious limitations for soybean production, with yield losses from 4 to 100%. The most effective method to reduce damage would be to grow Phytophthora-resistant soybean cultivars, and two types of host resistance have been described. Race-specific resistance conditioned by single dominant Rps ("resistance to Phytophthora sojae") genes and quantitatively inherited partial resistance conferred by multiple genes could both provide protection from the pathogen. Molecular markers linked to Rps genes or quantitative trait loci (QTLs) underlying partial resistance have been identified on several molecular linkage groups corresponding to chromosomes. These markers can be used to screen for Phytophthora-resistant plants rapidly and efficiently, and to combine multiple resistance genes in the same background. This paper reviews what is currently known about pathogenic races of P. sojae in the USA and Japan, selection of sources of Rps genes or minor genes providing partial resistance, and the current state and future scope of breeding Phytophthora-resistant soybean cultivars.

Variation in the Resistance of Japanese Soybean Cultivars to Phytophthora Root and Stem Rot during the Early Plant Growth Stages and the Effects of a Fungicide Seed Treatment.

Soybean cultivars susceptible to Phytophthora root and stem rot are vulnerable to seed rot and damping-off of seedlings and young plants following an infection by Phytophthora sojae. In this study, the disease responses of Japanese soybean cultivars including currently grown main cultivars during the early growth stages were investigated following infections by multiple P. sojae isolates from Japanese fields. The extent of the resistance to 17 P. sojae isolates after inoculations at 14, 21, and 28 days after seeding varied significantly among 18 Japanese and two US soybean cultivars. Moreover, the disease responses of each cultivar differed significantly depending on the P. sojae isolate and the plant age at inoculation. Additionally, the treatment of 'Nattosyo-ryu' seeds with three fungicidal agrochemicals provided significant protection from P. sojae when plants were inoculated at 14-28 days after seeding. These results indicate that none of the Japanese soybean cultivars are completely resistant to all tested P. sojae isolates during the first month after sowing. However, the severity of the disease was limited when plants were inoculated during the later growth stages. Furthermore, the protective effects of the tested agrochemicals were maintained for at least 28 days after the seed treatment. Japanese soybean cultivars susceptible to Phytophthora root and stem rot that are grown under environmental conditions favorable for P. sojae infections require the implementation of certain practices, such as seed treatments with appropriate agrochemicals, to ensure they are protected from P. sojae during the early part of the soybean growing season.

Thrombosed unruptured cerebral aneurysm causing brain infarction followed by subarachnoid hemorrhage--case report--.

A 71-year-old man presented with right hemiparesis and aphasia due to cerebral infarction in the frontal lobe. Computed tomography (CT) revealed a high-density mass, 12 mm in diameter, in the stem of the left sylvian fissure. Carotid angiography demonstrated occlusion of the left ascending frontal artery complex and retention of contrast medium at the bifurcation of the left middle cerebral artery (MCA). The diagnosis was cerebral infarction caused by occlusion of the ascending frontal artery complex resulting from thrombosed left MCA aneurysm. The patient was managed conservatively and his neurological symptoms gradually improved. One month later, he lapsed into a coma. CT revealed subarachnoid hemorrhage. Carotid angiography showed a large left MCA aneurysm with branch occlusion of the left ascending frontal artery complex. A left frontotemporal craniotomy was performed. The MCA aneurysm was opened and the intramural thrombi removed, and finally neck clipping was performed. The patient made a good postoperative recovery.

Hepatocyte growth factor promotes neuronal differentiation of neural stem cells derived from embryonic stem cells.

We previously reported that hepatocyte growth factor (HGF) promoted proliferation of neurospheres and neuronal differentiation of neural stem cells (NSCs) derived from mouse embryonic brain. In this study, spheres from mouse embryonic stem (ES) cells were generated by floating culture following co-culture on PA6 stromal cells. In contrast to the behavior of the neurospheres derived from embryonic brain, addition of HGF to the growth medium of the floating cultures decreased the number of spheres derived from ES cells. When spheres were stained using a MAP-2 antibody, more MAP-2-positive cells were observed in spheres cultured with HGF. When HGF was added to the growth and/or differentiation medium, more MAP-2-positive cells were also obtained. These results suggest that HGF promotes neuronal differentiation of NSCs derived from ES cells.

[Effect of an oral anti-estrogen agent (mepitiostane) on the regression of intracranial meningiomas in the elderly].

We report here the regression of meningioma following treatment with the anti-estrogen agent mepitiostane in a series of cases. The first case was that of a 72-year-old woman who presented with coma status due to non-communicating hydrocephalus. A large presumed meningioma within the cerebello-pontine angle was detected on gadolinium-enhanced magnetic resonance imaging (MRI). The patient recovered from the neurological deficit following endoscopic third ventriculostomy treatment, and was administered mepitiostane (10mg/day) orally. Gadolinium-enhanced MRI showed a marked regression (85%) of the meningioma following 60 months of oral medication. The second case was that of a 79-year-old woman with no neurological deficit; however, a presumed meningioma located in the frontal skull base was detected on gadolinium-enhanced MRI. Mepitiostane (10mg/day) was administered orally. Again, a marked regression (88%) of the meningioma was demonstrated after 115 months of oral medication. The third case was that of a 71-year-old woman who presented with right visual disturbance and a visual field defect. Gadolinium-enhanced MRI demonstrated a presumed meningioma located in the left sphenoidal bone. Mepitiostane (20mg/day) was administered orally. An 79% regression of the meningioma was observed after 21 months of oral medication. In these three cases, the marked reduction in meningioma following anti-estrogen agent (mepitiostane) administration suggested that this oral medication could be an effective therapeutic option in elderly patients.

Vertebral artery dissecting aneurysm with Cushing's syndrome: case report.

A 45-year-old woman presented with subarachnoid hemorrhage of World Federation of Neurosurgical Societies grade IV. Cerebral angiography showed a dissecting aneurysm of the right vertebral artery (VA). Internal trapping of the right VA with coils was performed. The postoperative course was uneventful, but she continued to demonstrate moon facies and experience amenorrhea. Computed tomography demonstrated an adrenal tumor. Laparoscopic adrenalectomy was performed under a diagnosis of Cushing's syndrome caused by an adrenal tumor. Overproduction of cortisol caused by Cushing's syndrome may be related to the development of cerebral aneurysm.

Hemispheric laminar necrosis as a complication of traumatic carotid-cavernous sinus fistula.

A 43-year-old woman presented with a very rare case of hemispheric laminar necrosis as a complication of traumatic carotid-cavernous sinus fistula (CCF). The patient suffered head injury and extensive burns following a car accident. Oral intubation was performed under sedation. When sedation was discontinued 17 days after injury, the patient demonstrated left hemiparesis. Magnetic resonance imaging showed laminar necrosis affecting the right cerebral hemisphere. Angiography revealed a right high-flow direct CCF. Transarterial embolization of the fistula using a detachable balloon achieved complete occlusion of the fistula. However, the left hemiparesis persisted following this intervention. Traumatic CCF may be missed in patients with disturbed consciousness, so clinicians should not overlook possibility of the triad of symptoms of CCF in patients with head injury.

Hemangioblastomatosis of the central nervous system without von Hippel-Lindau disease: a case report.

We report a very rare case of hemangioblastomatosis in a patient without von Hippel-Lindau disease (VHL). A 50-year-old woman had a history of surgical procedures for total removal of a cerebellar hemangioblastoma (HB). Twenty-one years after the last operation, she developed communicating hydrocephalus; computed tomographic (CT) scans of the brain showed no recurrence of HB in the posterior fossa. Subsequently, she underwent placement of a ventriculo-peritoneal shunt. One year later, she was readmitted because of progressive numbness and pain in the left lower limb. Magnetic resonance imaging (MRI) showed multiple Gd-enhancing tumors around the brain stem, in the cerebellum, and in the cervical and thoracolumbar regions of the spine. She underwent surgical removal of the tumors in the cerebellum and spinal cord. Although the extirpated tissues were histopathologically verified HB with less than 1% MIB-1 labeling index, surgery was followed by external beam radiation therapy with doses of 40 Gy to the whole brain, 10 Gy to the posterior fossa and 30 Gy to the whole spine. However, she subsequently developed quadriparesis and became bedridden.

Culture method for the induction of neurospheres from mouse embryonic stem cells by coculture with PA6 stromal cells.

Embryonic stem (ES) cells proliferate and maintain their pluripotency for over 1 year in vitro and may therefore provide a sufficient source for cell therapies. However, most of the previously reported methods for obtaining a source for cell therapies have not been simple. We describe here a novel method for induction of neurospheres from mouse ES cells by coculturing on PA6 cells instead of the formation of embryoid bodies. The ES cells cocultured with the PA6 stromal cell line for at least 3 days were capable of differentiating into spheres. The cells in the spheres were all green fluorescent protein (GFP) positive, showing that they were derived from GFP-expressing D3-ES cells. The spheres contained nestin-positive cells. The number of spheres increased when they were cocultured with PA6 for a longer period. Sphere formation was observed even after 10 mechanical dissociations and subculturings, showing its self-renewal ability. The cells differentiated into microtubule-associated protein-2 (MAP2)-positive neuronal cells and glial fibrillary acidic protein (GFAP)-positive glial cells. gamma-Aminobutyric acid-positive cells and tyrosine hydroxylase-positive cells were also observed in the spheres. The percentages of the MAP2- or GFAP-positive cells in the sphere changed according to the period of coculture on PA6 cells. At an early stage of coculture, more neurons were generated and, at a later period, more glial cells were generated. These results suggested that neurosphere could be generated from ES cells by coculturing with PA6, and that these cells resembled neural stem cells derived from mouse fetal brain tissue.

A non-infantile case of desmoplastic infantile astrocytoma.

CASE REPORT: We describe a very rare non-infantile case of desmoplastic infantile astrocytoma (DIA). A 9-year-old boy presented with motor weakness and sensory disturbance in his right upper and lower limbs. CT and MRI showed a contrast-enhanced large cystic tumor in the left sensorimotor area. We successfully resected the entire tumor. Its histopathological features were typical of DIA. OUTCOME: The patient's neurological symptoms improved postoperatively. Neither radiotherapy nor chemotherapy was used postoperatively. The patient developed normally and had been doing well for 12 months after surgery without tumor recurrence.