Developmental mechanisms regulating the formation of smooth muscle layers in the mouse uterus†.

Uterine smooth muscle cells differentiate from mesenchymal cells, and gap junctions connect the muscle cells in the myometrium. At the neonatal stage, a uterine smooth muscle layer is situated away from the epithelium when smooth muscle cells are grafted near the epithelium, suggesting that the epithelium plays an important role in differentiation, proliferation, and/or migration of smooth muscle cells. In this study, developmental mechanisms regulating the formation of the smooth muscle layers in the mouse uterus were analyzed using an in vitro culture model. Differentiation of smooth muscle cells occurs at a neonatal stage because ACTA2 gene expression was increased at the outer layer, and GJA1 was not expressed in cellular membranes of uterine smooth muscle cells by postnatal day 15. To analyze the effects of the epithelium on the differentiation of smooth muscle cells, a bulk uterine mesenchymal cell line was established from p53-/- mice at postnatal day 3 (P3US cells). Co-culture with Müllerian ductal epithelial cells (E1 cells) induced repulsive migration of ACTA2-positive cells among bulk P3US cells from E1 cells, but it had no effects on the migration of any of 100% ACTA2-positive or negative smooth muscle cell lines cloned from P3US cells. Thus, uterine epithelial cells indirectly affected the repulsive migration of smooth muscle cells via mesenchymal cells. Conditioned medium by E1 cells inhibited differentiation into smooth muscle cells of clonal cells established from P3US cells. Therefore, the uterine epithelium inhibits the differentiation of stem-like progenitor mesenchymal cells adjacent to the epithelium into smooth muscle cells.

The effect of electroconvulsive seizure on survival, neuronal differentiation, and expression of the maturation marker in the adult mouse hippocampus.

Electroconvulsive seizure (ECS), a model of electroconvulsive therapy in rodents, strongly increases neurogenesis in the adult hippocampus. Neurogenesis is a multi-step process that spans proliferation, survival, neuronal differentiation, and functional maturation. Our previous study demonstrated that ECS stimulates the proliferation of neural stem-like cells. However, the contribution of ECS to survival, neuronal differentiation, and maturation in newborn cells remains unknown. To evaluate the effect of ECS on these processes, we labeled newborn cells with bromodeoxyuridine (BrdU) before ECS treatment to determine the cell age and examined the survival rate and expression of cellular markers in the BrdU-labeled cells. Our results revealed that exposure to ECS (11 repetitions) during the differentiation phase significantly increased survival and promoted neuronal differentiation of newborn cells in the dentate gyrus. Four of ECS repetitions during the early differentiation phase were sufficient to promote dendritic outgrowth in immature neurons and enhance the expression of the immature neuronal marker, calretinin, in newborn cells. In contrast, exposure to ECS (11 repetitions) during the late maturation phase significantly suppressed the expression of the mature neuronal marker, calbindin, in newborn neurons. These results demonstrate that ECS during the differentiation phase promoted survival and neuronal differentiation and, in contrast, suppressed mature marker expression during the late maturation phase, suggesting that ECS has multiple effects on the different stages of adult neurogenesis.

Anaplastic changes of diffuse leptomeningeal glioneuronal tumor with polar spongioblastoma pattern.

Diffuse leptomeningeal glioneuronal tumor (DLGNT) is a rare glioneuronal neoplasm with oligodendroglioma-like cells confined in the subarachnoid spaces. A great majority of DLGNT are histologically low grade. However, some tumors show features of anaplasia with increased mitotic and proliferative activity. Due to the limited number of patients and inadequate clinical follow-up reported to date, the WHO classification does not yet assign a distinct WHO grade to this entity. Polar spongioblastoma pattern, in which bipolar cells are arranged in parallel with palisading nuclei, remains poorly understood about the pathological process of forming this pattern. We experienced a case of 22-year-old man developing DLGNT with extensively distributed anaplastic changes involving polar spongioblastoma pattern and the secondary tumor invasion to brain parenchyma in 4½ years before the autopsy. Clinical and pathological courses of the patient are presented with radiological, histopathological, and genetic examinations. This is the first report demonstrating the immunohistological and genetic evaluation of a DLGNT with polar spongioblastoma pattern.

The efficacy of resection of an intradural extramedullary foramen magnum cavernous malformation presenting with repeated subarachnoid hemorrhage: a case report.

BACKGROUND: Intradural extramedullary cavernous angiomas of the central nervous system are a rare type of cavernous angioma, but they can cause fatal subarachnoid hemorrhage. The efficacy of resection for this type of cavernous malformations remains uncertain. This is the first report to recommend surgical resection of these types of lesions regardless of the fatal condition. CASE PRESENTATION: Our patient was a 70-year-old Japanese man who experienced a sudden onset of an occipital headache, followed by bilateral abducens nerve palsy. Magnetic resonance imaging revealed a small amount of hemorrhage in both of the lateral ventricles and an intradural extramedullary mass lesion in the left side of his foramen magnum. Two weeks after the appearance of initial symptoms, he became comatose. A computed tomography scan showed an increase in the subarachnoid intraventricular hemorrhaging and of the acute hydrocephalus. Following ventricular drainage, total tumor resection was performed using the lateral suboccipital transcondylar approach in conjunction with a first cervical hemilaminectomy. We observed a grape-like vascular-rich tumor with calcification that was adhering tightly to the wall of his left vertebral artery. A histopathological examination of the surgery specimen identified it as a cavernous angioma. After placement of a ventriculoperitoneal shunt and 2 months of rehabilitation, he recovered completely. CONCLUSIONS: An intradural extramedullary foramen magnum cavernous malformation is quite rare. The fragile surface of our patient's lesion was causing repeated subarachnoid hemorrhage and consequently progressive fatal neurological deterioration. Surgical resection of the lesion to prevent repeated hemorrhage was performed and he recovered fully. Therefore, we recommend surgical resection of the lesion regardless of the potentially fatal condition.

Interferon-ß and temozolomide combination therapy for temozolomide monotherapy-refractory malignant gliomas.

Interferon-ß (IFN-ß) has been found to downregulate O6-methyl-guanine-DNA methyltransferase and sensitize glioma cells to chemoradiation therapy. The effectiveness of IFN-ß and temozolomide (TMZ) combination therapy for newly diagnosed glioblastomas was previously reported. However, there is no clinical report of recurrent of malignant gliomas treated with the combination of IFN-ß and TMZ. In the present study, we reported 7 cases of gliomas classified as uncontrollable with adjuvant TMZ monotherapy, who were then treated with IFN-ß and TMZ combination therapy. The magnetic resonance imaging findings and clinical symptoms improved in the majority of the cases, with tolerable adverse events and minimal residual disability. The overall survival (OS) time from the date of the initial surgery exceeded 13 months, suggesting that this combination therapy was successful in improving the prognosis of malignant gliomas refractory to adjuvant TMZ monotherapy.

Evaluation of the antiangiogenic effects of octreotide on growth hormone-producing pituitary adenoma using arterial spin-labeling perfusion imaging.

Decreased absolute tumor blood flow (TBF) measured by arterial spin labeling perfusion imaging (ASL-PI) on 3-tesla magnetic resonance imaging demonstrated the reduced size and growth hormone (GH) secretion of a large GH-producing pituitary adenoma in a 32-year-old man in response to octreotide therapy. The study shows the usefulness of ASL-PI in providing a biomarker of the antiangiogenic effect of octreotide.

Pseudo-continuous arterial spin labeling reflects vascular density and differentiates angiomatous meningiomas from non-angiomatous meningiomas.

Pseudo-continuous arterial spin labeling (PCASL) can measure tumor blood flow (TBF) reliably. We investigated meningioma TBF using PCASL and assessed for any correlation with histopathological microvascular density (MVD) and the World Health Organization (WHO) classification. Conventional MRI with contrast T1-weighted images and PCASL were acquired with a 3 T scanner before surgery in 25 consecutive patients with meningiomas. Using the PCASL perfusion map, the mean and maximum TBF were calculated from regions of interest placed in the largest cross-sectional plane of each tumor. Tissue sections from 16 patients were stained with CD31 to evaluate MVD and were assigned a WHO classification. The TBFs were statistically compared with MVD and the histopathological meningioma subtypes. There were 16 meningothelial meningiomas, four angiomatous meningiomas, two fibrous meningiomas, one transitional meningioma, and two atypical meningiomas. We observed significant correlation between MVD and both mean and maximum TBF (p < 0.05). The mean and maximum TBF ((mean)TBF, (max)TBF) in angiomatous meningiomas are significantly higher than that in non-angiomatous meningiomas (p < 0.05). PCASL is useful in assessing meningioma vascularity, and in differentiating angiomatous meningiomas from non-angiomatous meningiomas.

Nasopharyngeal carcinoma presenting with rapidly progressive severe visual disturbance: a case report.

INTRODUCTION: Nasopharyngeal carcinoma is one of the most difficult tumors to diagnose correctly at the initial phase because of the occasional lack of nasal symptoms. The perineural spread of the trigeminal nerve is one of the most common and important routes in the intracranial paracavernous extension of nasopharyngeal carcinoma, but visual loss is very rare. CASE PRESENTATION: We report the case of a 54-year-old Japanese man with nasopharyngeal carcinoma, who presented with rapid and severe disturbance of left monocular visual acuity and eye movement with a 10-month history of ipsilateral otitis media and facial pain. Magnetic resonance imaging revealed a lesion in the left fossa of Rosenmüller, pterygopalatine fossa, sphenoid and ethmoid sinus, and the left cavernous sinus extending to the orbital apex through the superior orbital fissure. The histopathological diagnosis was nonkeratinizing undifferentiated nasopharyngeal carcinoma. Epstein-Barr virus was detected by in situ hybridization. Although focal radiotherapy induced remarkable tumor shrinkage and relieved ocular motor disturbance and facial pain, his visual acuity did not improve. CONCLUSION: The awareness of cranial nerves in addition to intracranial and orbital apex involvement, as in this case, is important for appropriate diagnosis and treatment planning of nasopharyngeal carcinoma.

Primary neurolymphomatosis of the lower cranial nerves presenting as Dysphagia and hoarseness: a case report.

Primary neurolymphomatosis is an extremely rare tumor. We report the case of a 74-year-old patient presenting with dysphagia and hoarseness. Initial contrast-enhanced computed tomography of the head, neck, and chest did not reveal any lesions. His symptoms improved with short-term administration of prednisone but recurred and deteriorated. Magnetic resonance (MR) imaging revealed a tumor along the ninth and tenth cranial nerves across the jugular foramen. Fluorine-18 fluorodeoxyglucose positron emission tomography indicated this was a primary tumor. Repeated MR imaging after 2 months revealed considerable tumor enlargement. A left suboccipital craniotomy was performed to remove the tumor that infiltrated the ninth and tenth cranial nerves. The histopathologic diagnosis was diffuse large B-cell lymphoma. Although focal radiation therapy was administered to ensure complete eradication of the tumor, the patient died of aspiration pneumonia with systemic metastasis. To our knowledge, this is the first reported case of primary neurolymphomatosis in the lower cranial nerves.

Usefulness of intraoperative monitoring of visual evoked potentials in transsphenoidal surgery.

Postoperative visual outcome is a major concern in transsphenoidal surgery (TSS). Intraoperative visual evoked potential (VEP) monitoring has been reported to have little usefulness in predicting postoperative visual outcome. To re-evaluate its usefulness, we adapted a high-power light-stimulating device with electroretinography (ERG) to ascertain retinal light stimulation. Intraoperative VEP monitoring was conducted in TSSs in 33 consecutive patients with sellar and parasellar tumors under total venous anesthesia. The detectability rates of N75, P100, and N135 were 94.0%, 85.0%, and 79.0%, respectively. The mean latencies and amplitudes of N75, P100, and N135 were 76.8 ± 6.4 msec and 4.6 ± 1.8 µV, 98.0 ± 8.6 msec and 5.0 ± 3.4 µV, and 122.1 ± 16.3 msec and 5.7 ± 2.8 µV, respectively. The amplitude was defined as the voltage difference from N75 to P100 or P100 to N135. The criterion for amplitude changes was defined as a > 50% increase or 50% decrease in amplitude compared to the control level. The surgeon was immediately alerted when the VEP changed beyond these thresholds, and the surgical manipulations were stopped until the VEP recovered. Among the 28 cases with evaluable VEP recordings, the VEP amplitudes were stable in 23 cases and transiently decreased in 4 cases. In these 4 cases, no postoperative vision deterioration was observed. One patient, whose VEP amplitude decreased without subsequent recovery, developed vision deterioration. Intraoperative VEP monitoring with ERG to ascertain retinal light stimulation by the new stimulus device was reliable and feasible in preserving visual function in patients undergoing TSS.

Usefulness of intraoperative monitoring of visual evoked potentials in transsphenoidal surgery.

Postoperative visual outcome is a major concern in transsphenoidal surgery (TSS). Intraoperative visual evoked potential (VEP) monitoring has been reported to have little usefulness in predicting postoperative visual outcome. To re-evaluate its usefulness, we adapted a high-power light-stimulating device with electroretinography (ERG) to ascertain retinal light stimulation. Intraoperative VEP monitoring was conducted in TSSs in 33 consecutive patients with sellar and parasellar tumors under total venous anesthesia. The detectability rates of N75, P100, and N135 were 94.0%, 85.0%, and 79.0%, respectively. The mean latencies and amplitudes of N75, P100, and N135 were 76.8±6.4 msec and 4.6±1.8 µV, 98.0±8.6 msec and 5.0±3.4 µV, and 122.1±16.3 msec and 5.7±2.8 µV, respectively. The amplitude was defined as the voltage difference from N75 to P100 or P100 to N135. The criterion for amplitude changes was defined as a >50% increase or 50% decrease in amplitude compared to the control level. The surgeon was immediately alerted when the VEP changed beyond these thresholds, and the surgical manipulations were stopped until the VEP recovered. Among the 28 cases with evaluable VEP recordings, the VEP amplitudes were stable in 23 cases and transiently decreased in 4 cases. In these 4 cases, no postoperative vision deterioration was observed. One patient, whose VEP amplitude decreased without subsequent recovery, developed vision deterioration. Intraoperative VEP monitoring with ERG to ascertain retinal light stimulation by the new stimulus device was reliable and feasible in preserving visual function in patients undergoing TSS.

Dissecting Aneurysm at the Proximal Segment of the Anterior Cerebral Artery Associated with Infraoptic Course Anterior Cerebral Artery.

A 48-year-old man presented a subarachnoid hemorrhage caused by a rupture of a dissecting aneurysm at the proximal segment (A1 segment) of the right anterior cerebral artery (ACA). He also had an anomalous artery named infraoptic course ACA and an agenesis of the contralateral ACA A1 segment. Balloon occlusion test at the bifurcation of the right internal carotid artery demonstrated that the distal segments of the bilateral ACAs were perfused through the infraoptic course ACA. Therefore, we surgically trapped the A1 segment including the aneurysm. The patient got discharged without any neurological deficit. Natural course of ACA dissecting aneurysms is unclear because of rarity of the disease and treatment strategy is still controversial. Most of the dissecting aneurysms in the A1 segment are surgically treated, because they often present with massive hemorrhage and poor prognosis. In the present case, the contralateral A1 segment was absent but trapping of the dissecting aneurysm could be achieved without vascular reconstruction (e.g., bypass surgery) because of the presence of the infraoptic course ACA.

Evaluation of tumor blood flow after feeder embolization in meningiomas by arterial spin-labeling perfusion magnetic resonance imaging.

Preoperative embolization changes the amount of blood flow and pattern of flow distribution in meningioma. Tumor blood flow was investigated in eight meningioma patients before and after embolization using arterial spin-labeling (ASL) perfusion imaging. Although blood flow was significantly reduced in the whole tumor after embolization, changes in flow distribution patterns varied from one case to another. The findings suggest that evaluation of post-embolization tumor blood flow by ASL perfusion imaging would be useful in the surgical planning of meningioma.

Cerebral hemorrhage due to posterior reversible encephalopathy syndrome associated with autonomic dysreflexia in a spinal cord injury patient.

A 37-year-old man with C4-5 spinal cord injury (SCI) presented with abnormally high blood pressure after vesicocutaneous catheter exchange and was treated with antihypertensive agents. Two weeks later, he developed headache and visual disturbance, and presented with fluctuating blood pressure. Multiple subcortical hemorrhages in the left occipital and right frontal lobes occurred on the next day, and he died of increased intracranial pressure 3 weeks later. Based on the symptoms and computed tomography findings, the retrospective diagnosis was posterior reversible encephalopathy syndrome (PRES) due to autonomic dysreflexia (AD). AD occurs frequently in patients with quadriplegia and high paraplegia by distention of the bladder and bowel. PRES secondary to AD is very rare, but we must always be aware of this life-threatening complication in SCI patients.

Chronic subdural hematoma in a patient with congenital afibrinogenemia successfully treated with fibrinogen replacement.

A 37-year-old woman with congenital afibrinogenemia presented with chronic subdural hematoma (CSDH) manifesting as severe headache, nausea, and somnolence after a minor head trauma. Brain computed tomography scans showed a right subdural hematoma associated with midline shift. Laboratory studies showed prolongation of prothrombin time, activated partial thromboplastin time, and undetectably low level of fibrinogen. Until the present episode, she had received plasma-derived fibrinogen concentrate around menstruation and pregnancy. She had also suffered from spinal cord infarction due to vertebral artery occlusion. Burr-hole evacuation and drainage of CSDH was successfully performed using fibrinogen concentrate. The development of CSDH with afibrinogenemia is very rare. Although the past repeated administrations of fibrinogen concentrate were suspected to generate CSDH, paradoxical thrombotic complications caused by upregulation of prothrombin activation, thrombin generation, and growth factors released from platelets might be related to the development of CSDH with congenital afibrinogenemia.

Basi-parallel anatomical scanning magnetic resonance imaging in patients with bilateral vertebrobasilar artery dissections.

A 31-year-old man presented with bilateral unruptured vertebral artery (VA) dissections. The temporal course of the dissected arteries was successively evaluated using brain surface imaging modality basi-parallel anatomical scanning (BPAS) magnetic resonance (MR) imaging in combination with the conventional modalities of MR angiography and three-dimensional computed tomography (3D-CT) angiography. Initially, BPAS-MR imaging clearly demonstrated bilateral fusiform dilatations of the arterial wall, whereas MR angiography and 3D-CT angiography showed irregular, dilated, or interrupted inner contour of the arteries. Finally, BPAS-MR imaging demonstrated resolution of both aneurysms, and MR angiography demonstrated obstruction of the left VA and normal contours of the right VA. Combination of diagnostic tools such as BPAS-MR imaging and MR angiography is useful for evaluating the exact nature of dissected arteries and determining the temporal course.

Mixed germ cell tumor and hemangioblastoma in the cerebellum: report of a rare coexistence.

We report a case of a cerebellar tumor consisting of a mixed germ cell tumor (GCT) and a hemangioblastoma. A 22-year-old man presented with myoclonus and cerebellar ataxia. Magnetic resonance imaging showed a tumor mass in the left cerebellar hemisphere. The tumor was totally removed, and the histological diagnosis was an undetermined neoplasm. Ten months later, the patient returned with cerebellar hemorrhage at the site of the previous tumor. An emergency craniotomy was performed, and a tumor mass adjacent to the hematoma was resected. Microscopic examination revealed a mixed GCT consisting of a germinoma, choriocarcinoma, and mature teratomatous component. An area of hemangioblastoma was also found in the same tumor mass. A retrospective examination of the histological sample from the first operation indicated a germinoma. A primary GCT of the posterior fossa is very rare, and there are no other reports of the coexistence of a GCT and a hemangioblastoma. A metastatic GCT lesion of extracranial origin should be considered when the intracranial GCT is non-germinomatous and arises in an unusual site. The most probable hypothesis for the histogenesis of this case was a hemangioblastoma complicated by a "tumor-to-tumor" metastatic lesion of testicular GCT with "burnout" of the primary site.

Enhancement of antibody production by the addition of Coenzyme-Q(10).

Recently, there has been a growing demand for therapeutic monoclonal antibodies (MAbs) on the global market. Because therapeutic MAbs are more expensive than low-molecular-weight drugs, there have been strong demands to lower their production costs. Therefore, efficient methods to minimize the cost of goods are currently active areas of research. We have screened several enhancers of specific MAb production rate (SPR) using a YB2/0 cell line and found that coenzyme-Q(10) (CoQ(10)) is a promising enhancer candidate. CoQ(10) is well known as a strong antioxidant in the respiratory chain and is used for healthcare and other applications. Because CoQ(10) is negligibly water soluble, most studies are limited by low concentrations. We added CoQ(10) to a culture medium as dispersed nanoparticles at several concentrations (Q-Media) and conducted a fed-batch culture. Although the Q-Media had no effect on cumulative viable cell density, it enhanced SPR by 29%. In addition, the Q-Media had no effect on the binding or cytotoxic activity of MAbs. Q-Media also enhanced SPR with CHO and NS0 cell lines by 30%. These observations suggest that CoQ(10) serves as a powerful aid in the production of MAbs by enhancing SPR without changing the characteristics of cell growth, or adversely affecting the quality or biological activity of MAbs.

Comparison of cell lines for stable production of fucose-negative antibodies with enhanced ADCC.

Several methods have been described to enhance antibody-dependent cellular cytotoxicity (ADCC) using different host cells that produce antibody with reduced levels of fucose on their carbohydrates. We compared the suitability of these methods for the serum-free fed-batch production of antibody for clinical trials and commercial uses. Recombinant anti-human CD20 chimeric IgG1-producing clones were established from host-cells that have been shown to produce more than 90% fucose-negative antibody. The cell lines were a FUT8 (alpha-1,6-fucosyltransferase) knockout Chinese hamster ovary (CHO) cell line, Ms704, and two Lens culinaris agglutinin (LCA)-resistant cell lines, one derived from a variant CHO line, Lec13 and the other from a rat hybridoma cell line, YB2/0. The amount of fucose-negative antibody produced by Lec13 and YB2/0 significantly decreased with the culture. The increase in fucosylation was due to remaining synthesis of GDP-fucose via de novo pathway for the CHO line and the elevation of FUT8 expression by the YB2/0 cells. In contrast, Ms704 cells stably produced fucose-negative antibody with a consistent carbohydrate structure until the end of the culture. The productivity of the Ms704 cells reached 1.76 g/L with a specific production rate (SPR) of 29 pg/cell/day for 17 days in serum-free fed-batch culture using a 1 L spinner bioreactor. Our results demonstrate that FUT8 knockout has the essential characteristics of host cells for robust manufacture of fucose-negative therapeutic antibodies with enhanced ADCC.