Amniotic epithelial cells transform into neuron-like cells in the ischemic brain.

We investigated the potential use of rat amniotic epithelial (RAE) cells as donor cells for transplantation-based therapy in brain ischemia. In vitro, RAE cells were positive for both neuronal and neural stem cell markers, neurofilament microtubule-associated protein 2 and nestin. RT-PCR revealed that these cells express nestin mRNA. The RAE cells were transplanted into the hippocampus of adult gerbils that were subjected to temporal occlusion of bilateral carotid arteries. Five weeks after transplantation, grafted cells migrated into the CA1 pyramidal layer that showed selective neuronal death, and survived in a manner similar to CA1 pyramidal neurons. These results suggest that intracerebral transplantation of amniotic epithelial cells may have therapeutic potential for the treatment of ischemic damage in neuronal disorders.

Preparation and clinical application of 2% diflunisal oral ointment for painful lesions of the oral mucosa.

We previously reported the development and clinical efficacy of a 2% aspirin oral ointment and 2% ethenzamide oral ointment as hospital preparations for painful lesions of the oral mucosa. This study investigated methods of preparing a more stable oral ointment with a more effective analgesic action, using diflunisal, another salicylic acid derivative, with an analgesic effect stronger than that of aspirin. A two-percent diflunisal oral ointment was prepared similarly to the aspirin ointment using plastibase and CMC-Na as the ointment base. From the results of spreadability measurement, a CMC-Na content of 20% was considered appropriate. The stability of diflunisal in 2% diflunisal oral ointment stored at 5 degrees C, 20 degrees C and 30 degrees C, was determined using HPLC, and a high stability of diflunisal at room temperature for more than 100 days was confirmed. We also investigated its antinociceptive effect using the Randall-Selitto paw pressure test in rats, which showed that 2% diflunisal oral ointment was as effective as 2% aspirin oral ointment. On clinical application of 2% diflunisal oral ointment to 8 patients with painful oral mucous diseases, it was found to be significantly (p = 0.014) more effective than 2% aspirin oral ointment. The results of this study demonstrated that 2% diflunisal oral ointment is a clinically useful analgesic for painful oral lesions.

Surgical experience of 120 patients with lumbosacral lipomas.

The surgical experience of 120 patients with lumbosacral lipomas is described. 47 of 120 patients were preoperatively found to be neurologically intact, the remaining 73 patients presented with various neurological signs including reflex changes, sensory disturbances, muscle weakness and sphincter problems. Neuro-imagings allowed a classification of lumbosacral lipomas into five types: (1) dorsal type; (2) caudal type; (3) combined type; (4) filar type; and (5) lipomyelomeningocele. Although all 120 patients underwent untethering of the spinal cord, the nerve roots passing through the lipoma itself and the neural tissues protruding externally to the spinal canal, respectively, tended to prevent satisfactory surgical removal of the lipoma in combined type lipomas and lipomyelomeningoceles. During 8.96 years of a mean postoperative follow-up period, there was no significant deterioration in most of the patients and some patients even improved in function. However, two patients with combined type lipomas developed neurological deterioration just after surgery, and five (two dorsal, two caudal and one combined type lipomas) did in the fashion of a late-onset. There are two different patient groups of lumbosacral lipomas; one group (caudal and filar type lipomas, and most of dorsal type lipomas) in whom the surgical anatomy is simple and satisfactory untethering surgery could be done without risk, and another (combined type lipomas and lipomyelomeningocele) in whom surgery would be accompanied with some risk and sometimes complete untethering could not be achieved because of the complicated anatomy of the lesion. Surgical difficulty of the latter group can be correlated with the increased frequency of neurological deterioration occurring just after the operation, but not of delayed one. Concerning prophylactic surgery for asymptomatic patients, the former group of patients are obviously good candidates, but the latter group is not.

Engraftment of genetically engineered amniotic epithelial cells corrects lysosomal storage in multiple areas of the brain in mucopolysaccharidosis type VII mice.

Cell-mediated gene therapy for visceral lesions of lysosomal storage diseases is promising; however, the treatment of central nervous system (CNS) lesions remains a challenge. In this study, we generated rat amniotic epithelial cells (AEC) that overexpress and secrete human beta-glucuronidase (GUSB) following transduction with an adenoviral vector encoding human GUSB. The AEC were used as donor cells for cell-mediated gene therapy of CNS lesions in mice with mucopolysaccharidosis type VII (MPSVII), a lysosomal storage disorder caused by an inherited deficiency of GUSB activity. After confirmation that the secreted GUSB was taken up mainly via mannose 6-phosphate receptors in primary cultured neurons, the AEC were transplanted into the brains of adult MPSVII mice. Histochemical analysis showed extensive GUSB activity throughout the ipsilateral hemisphere of the recipient brains, and pathological improvement of the lysosomal storage was observed even in regions far from the site of injection. These results suggest that intracerebral transplantation of genetically engineered AEC has therapeutic potential for the treatment of CNS lesions in lysosomal storage disorders.

Possible origin of suprasellar arachnoid cysts: neuroimaging and neurosurgical observations in nine cases.

OBJECT: In this study the authors identify and investigate two new classifications of suprasellar arachnoid cysts. METHODS: The authors used computerized tomography cisternography, magnetic resonance (MR) imaging, and neuroendoscopy to investigate nine cases of suprasellar arachnoid cysts. A communicating cyst with early filling and early clearance of a radioopaque tracer was found in seven of nine cases; a communicating cyst with delayed filling and delayed clearance of the tracer was observed in one case; and a noncommunicating cyst was observed in the other. The MR findings indicated a variation in the position of the basilar artery (BA) bifurcation in relation to the ventral surface of the midbrain. A distance existed between the BA bifurcation and the ventral surface of the midbrain in a communicating cyst with early filling, whereas the BA bifurcation was posteriorly displaced in a communicating cyst with delayed filling and also in a noncommunicating cyst, leaving little space between the bifurcation and the ventral surface of the midbrain. Endoscopic observation revealed, in the case of communicating cysts with early filling and early clearance of tracer, that the BA bifurcation is located inside the cyst with no overlying membrane, whereas in a noncommunicating cyst, the BA and its branches can be observed through the transparent membrane of the lesion. CONCLUSIONS: The authors postulate two different types of suprasellar arachnoid cysts: a noncommunicating intraarachnoid cyst of the diencephalic membrane of Liliequist and a communicating cyst that is a cystic dilation of the interpeduncular cistern.

Early experience with poly L-lactic acid bioabsorbable fixation system for paediatric craniosynostosis surgery. Report of 3 cases.

OBJECTIVE: The authors describe early experience in the use of bioabsorbable fixation devices for cranial reconstruction of paediatric craniosynostosis patients. METHODS: Three patients, individually respectively presenting sagittal synostosis, metopic synostosis, and clover leaf skull deformity, underwent cranial reconstruction using poly L-lactic acid (PLLA) plates and screws. The patients ranged in age from 2 to 10 months at the time of surgery. The postoperative clinical follow-up ranged from 16 to 18 months. All patients showed satisfactory wound healing without signs of infection or local inflammation. The plates provided satisfactory fixation and were not palpable through the skin after 16 to 18 postoperative months. CONCLUSION: Our experience demonstrated the efficacy of PLLA bioabsorbable plates and screws for cranial reconstruction in cases of infant craniosynostosis. Prospective studies and longer follow-up of a larger number of patients is desirable for confirmation of these findings.

The delivery of protein into living cells by use of membrane fusible erythrocyte ghosts.

We examined the effects of pretreatment of monkey kidney normal cells (CV-1) and human cervical tumor cells (HeLa) with membrane fusible erythrocyte ghosts (MFEG) encapsulating superoxide dismutase (SOD-MFEG) on adriamycin (ADM) cytotoxicity. The decrease in CV-1 cells with ADM was inhibited markedly, and the initiation of decrease in HeLa cells was delayed for 2 days by the pretreatment with SOD-MFEG, indicating that the delivered SOD held the activity in the cells.

Effects of incorporation of various amphiphiles into recipient liposome membranes on inter-membrane protein transfer.

To obtain information about the factors governing spontaneous inter-membrane protein transfer, we examined the effects of incorporation of various amphiphilic compounds in dimyristoylphosphatidylcholine (DMPC) liposomes on protein transfer from influenza virus-infected cells to the liposomes, and analyzed the physical properties of these liposome membranes. The incorporation of amphiphilic compounds, negatively charged dicetylphosphate (DCP), dipalmitoylphosphatidylserine (DPPS) or positively charged dimethyldipalmitoylammonium (DMDPA), into DMPC liposomal membranes enhanced protein transfer. The liposomes containing DCP, DPPS or DMDPA were unaffected by osmotic shock caused by external addition of glucose, suggesting a decrease in lipid packing in the liposomal membranes. Furthermore, calorimetric study of these liposomes showed that a phase separation occurred partially in the liposomal membranes. Accordingly, the membranes of DMPC liposomes containing DCP, DPPS and DMDPA should be distorted due to the coexistence of two phases, gel and liquid crystalline, in the membranes. Consequently, the membrane distortion could be responsible for the enhancement effects of the amphiphiles on the inter-membrane protein transfer from influenza virus-infected cells to the liposomes.

Successful surgical treatment of a dural arteriovenous fistula at the craniocervical junction with reference to pre- and postoperative MRI.

We report a 62-year-old woman who presented with a myelopathy at the lower thoracic level. Left vertebral angiography revealed a dural arteriovenous fistula (DAVF) at the craniocervical junction, draining into an anterior spinal vein. Below the T7 level, the spinal cord gave high signal on T2-weighted images and enhanced with Gd-DTPA. The patient was successfully treated by simple clipping of vein draining the DAVF. The abnormal signal intensity and contrast enhancement rapidly regressed, except in the conus medullaris. Regression of the parenchymal abnormality on serial MRI following treatment corresponded closely with postoperative improvement of neurological function.

Microanatomy of the cerebellopontine angle and internal auditory canal: study with new magnetic resonance imaging technique using three-dimensional fast spin echo.

OBJECTIVE: We report a new magnetic resonance imaging technique that uses three-dimensional fast spin echo and the minimum intensity projection method. Using this technique, detailed images of the cerebellopontine angle (CPA) and internal auditory canal (IAC) were obtained in normal volunteers and in patients with acoustic neuromas or hemifacial spasm. METHODS: Ten normal volunteers, 44 patients with acoustic neuromas, and 31 patients with hemifacial spasm were studied using the three-dimensional fast spin echo magnetic resonance imaging protocol. The CPA and IAC were scanned by using a 1-mm slice thickness in the axial and parasagittal planes. RESULTS: Normal anatomy was as follows. 1) The vestibulocochlear nerve was ovoid near the brain stem and changed to a slightly crescentic configuration (C shape) as it traveled laterally. 2) Separation of the cochlear and vestibular nerves was observed near the central part of the IAC. 3) Discrimination between the superior and inferior vestibular nerves was also possible near the fundus of the IAC. 4) The facial nerve was easily identifiable as a discrete nerve at the anterior aspect of the vestibulocochlear nerve. 5) The meatal loop of the cerebellar artery was located medial to the porus in 44% of 95 CPAs and reached the porus or protruded into the porus in 56%. Acoustic neuromas were as follows. 1) In a patient with a very small intracanalicular tumor, the nerve on which the tumor was located could be identified. 2) In 22 of 44 acoustic neuromas, cerebrospinal fluid was present between the tumor and the fundus of the IAC. Hemifacial spasm was as follows. The relationship between the responsible artery and the facial nerve could be precisely observed. CONCLUSION: The three-dimensional fast spin echo method offers ultrahigh-resolution images, which are extremely useful in understanding the surgical anatomy of the CPA and IAC.

Delineation of small nerves and blood vessels with three-dimensional fast spin-echo MR imaging: comparison of presurgical and surgical findings in patients with hemifacial spasm.

BACKGROUND AND PURPOSE: We applied a 3D fast spin-echo (3D-FSE) MR imaging technique to the preoperative and postoperative evaluation of patients with hemifacial spasm. METHODS: The study group comprised 20 patients. All images were acquired on a 1.5-T MR system with a 3D-FSE sequence. RESULTS: In all 20 patients, the courses of the seventh and eighth cranial nerves were depicted separately, and the arteries presumed to be responsible for the hemifacial spasm were seen to be in contact with the facial nerves at the root exit zone (REZ). Eight patients underwent neurovascular decompression. In all patients, the presumed responsible blood vessels depicted by 3D-FSE MR imaging corresponded to intraoperative findings. In addition, postoperative 3D-FSE images confirmed the separation of the facial nerve from a contiguous vessel at the REZ. DISCUSSION: The 3D-FSE technique makes it possible to obtain extremely high-quality images of microstructures in the cerebellopontine cistern, and it has several advantages over conventional angiography: it is noninvasive and able to depict the cranial nerves and surrounding vessels in the same image without contrast material, and it may be useful for postoperative evaluation of the decompression procedure. This imaging technique is expected to prove useful for the clinical evaluation of hemifacial spasm.

Effect of dicetylphosphate or stearic acid on spontaneous transfer of protein from influenza virus-infected cells to dimyristoylphosphatidylcholine liposomes.

Membrane proteins, such as viral spike, were transferred spontaneously from influenza virus-infected cells to various liposomes. The protein transfer was enhanced by the presence of negative charged component dicetylphosphate (DCP) or stearic acid (SA) in dimyristoylphosphatidylcholine (DMPC) liposomes. The lowering of membrane fluidity did not relate to the effect of DCP or SA on protein transfer in this study. We considered that the alteration of membrane properties, such as construction of the surface or stability of transferred protein in liposomes, due to the specific structure of DCP or SA is responsible for the enhancement of spontaneous protein transfer by the presence of the amphiphilic components.

Evidence for active acetylcholine metabolism in human amniotic epithelial cells: applicable to intracerebral allografting for neurologic disease.

Human amniotic epithelial (HAE) cells have been used for allotransplantation in patients with lysosomal storage disease due to lack of expression of HLA antigens. Previously, we have reported the expression of differentiation markers for both neural stem cells, and neuron and glial cells. In the present study, we investigated the presence of choline acetyltransferase (ChAT) and acetylcholine (ACh) in HAE cells using different experimental approaches. Cultured HAE cells showed strong immunoreactivity against ChAT antibody. ChAT activity in primary cells was 24.9 +/- 8.5 pmol/mg protein/h. Using HPLC with electrochemical detection, ACh was detected in both cell incubation media and cell pellets indicating that these cells synthesize and release ACh in a time-dependent manner. Additional confirmation of this hypothesis was gained from the data obtained from RT-PCR and Western blot analyses which revealed the expression of ChAT mRNA and ChAT protein, respectively, in HAE cells. Results of the present study suggest that HAE cells can possibly be applied for intracerebral allografting to treat neurologic diseases in which cholinergic neurons are damaged.

Development of a membrane fusible drug carrier from erythrocytes by the spontaneous transfer of viral fusion protein from influenza virus-infected cells.

In order to develop a membrane fusible drug carrier from human erythrocytes, we attempted the reconstitution of influenza virus fusion protein hemagglutinin (HA) to an erythrocyte membrane. In this study, we succeeded in the preparation of HA-reconstituted erythrocytes (HA-erythrocytes) by the incubation of erythrocytes with influenza virus-infected CV-1 cells, and confirmed the ability of HA-erythrocytes to fuse with the cell membrane. Furthermore, by using an HA-reconstituted ghost (HA-ghost), which entrapped fluorescent-labeled ovalbumin, 25% of the protein was incorporated into cells through the fusion of the HA-ghost with the cell membrane.

MR height of the pituitary gland as a function of age and sex: especially physiological hypertrophy in adolescence and in climacterium.

PURPOSE: To clarify the age- and sex-related changes in pituitary height. METHODS: We reviewed sagittal T1-weighted MR images in 1020 subjects to analyze the size of the pituitary gland. RESULTS: The pituitary height in female subjects (mean, 5.35 mm; SD, 1.2) was significantly greater than that in male subjects (mean, 4.93 mm; SD, 1.0). When the data were analyzed for different age ranges, sex-related differences were statistically significant only in the 10- to 19-, 20- to 29-, and 50- to 59-year-old age groups. The pituitary height peaked in the 20-to-29 age group and tended to decline with age. In female subjects, however, there was a tendency for pituitary height to increase again in the 50-to-59 age group. CONCLUSION: The pituitary height may reflect physiological neuroendocrine differences between younger and older subjects and between male and female subjects. The second increase observed in female subjects may likewise reflect high concentrations of gonadotropic hormones, apparently due to an age-related decline in circulating gonadal steroids and an increase in gonadotropin-releasing hormone.

Arachnoid cysts of the middle cranial fossa: experience with 77 patients who were treated with cystoperitoneal shunting.

OBJECTIVE: Our surgical results of middle fossa arachnoid cysts are presented, and advantages and disadvantages of cystoperitoneal shunting are discussed. METHODS: Seventy-seven patients with middle fossa arachnoid cysts, including 20 cases of asymptomatic cyst, were treated by cystoperitoneal shunting. In cases associated with subdural hematoma, the cystoperitoneal shunt was preceded by irrigation of the hematoma. Follow-up periods ranged from 8 months to 16 years (mean, 7.7 yr). RESULTS: In all cases, postoperative computed tomography and magnetic resonance imaging revealed subsequent expansion of the surrounding brain, concomitant with either the disappearance of or a marked decrease in the size of the cyst. Patients who initially presented with headaches, hemiparesis, choked discs, and diplopia were cured postoperatively. Head enlargement presented preoperatively did not progress after shunting. Mental retardation and behavior problems, however, remained unchanged postoperatively, even though the cyst had disappeared. One cystoperitoneal shunt became infected, and shunt malfunction developed in eight patients. Four of the eight patients presented acute signs and symptoms of intracranial hypertension when the shunt malfunctioned, although the concurrent increase in cyst size was slight and ventriculomegaly was not seen. The observed condition in these four patients could be considered shunt dependency. CONCLUSION: Cystoperitoneal shunting is a valid method of achieving obliteration of the cyst and clinical improvement and seems to feature low operative risk and few complications. It should be noted, however, that some patients experience shunt dependency after cystoperitoneal shunting.

[A clinicopathological study of 21 cases of primary central nervous system lymphoma].

A clinico-pathological study was carried out in 21 cases of primary central nervous system-non-Hodgkin's lymphoma (CNS-NHL). Their clinical profiles (age, prognosis, modalities of treatment) and findings of radio-imaging were analyzed. All specimens from surgery and/or autopsy were histologically classified according to the working formulation (WF) classification of the National Cancer Institute. Ontogeny of lymphoma cells was determined by immunohistochemical study in all cases and some cases were subjected to light (kappa, lambda) and heavy chain (IgG, IgA, IgM) analysis as well. Among 21 cases, 12 cases were located in the cerebral hemisphere, 7 in the thalamus-basal ganglia and 4 in the cerebellum. Radio-imaging study showed that 18 cases (86%) revealed isodensity mass lesions on plain CT, which were homogeneously enhanced by contrast medium. The pathological study showed that all cases were derived from B-cells. Five were classified as immunoblastic type (IBL), 9 as diffuse large type (DL), and the others were classified according to WF. 17 of 21 cases (81%) were sensitive to radiotherapy, and 15 of 19 cases (79%) responded to corticosteroid. A prognostic study revealed that patients with IBL had less hope than those with DL. From this result, it seems that WF classification is better than LSG classification for obtaining a prognosis in malignant lymphoma patients. The frequency of primary CNS-NHL has been increasing for the past several decades and will surpass that of any other brain tumors in the near future because of the explosive expansion of AIDS patients. Therefore, not only clinicopathological analysis but also biological study for CNS-NHL might be important.

Pituitary apoplexy caused by endocrine stimulation tests: a case report.

The case of a 60-year-old woman with a giant pituitary adenoma who suffered symptomatic pituitary apoplexy is reported. Hemorrhaging occurred during endocrine stimulation tests (triple bolus test) in and around the tumor. Both the level of consciousness and the hemiparesis improved after emergency decompression. Histologically the tumor was a chromophobe adenoma of the papillary type with diffuse hemorrhage. Subtotal removal of the tumor was successfully accomplished in a second operation. Routine anterior pituitary stimulation tests have the potential to develop pituitary apoplexy, and we advise that caution should be taken when such tests are planned. The possible mechanism resulting in bleeding is discussed.

Inverted Bruns' nystagmus in arachnoid cysts of the cerebellopontine angle.

A 9-year-old girl with huge, double arachnoid cysts at the cerebellopontine angle is reported. After a cyst-peritoneal shunt operation, she developed fine, high-frequency gaze nystagmus towards the lesion side combined with coarse, large amplitude nystagmus in the opposite direction, which is the reverse of the Bruns' nystagmus. We call this reverse phenomenon 'inverted' Bruns' nystagmus. As the nystagmus disappeared after a second shunt operation, the nystagmus was presumed to be caused by the cystic lesion.

Viral Kirsten ras infection differentiates PC12 cells and enhances their survival upon implantation into brain.

Neuronal cells from established cell lines can offer a well-characterized source of cells for transplantation to the brain that is an alternative to fetal neurons. The infection of members of the PC12 cell line with a retrovirus containing ras-oncogene leads to their neuronal differentiation without the need of nerve growth factor (NGF). We find that neoplastic, naive PC12 cells grafted to the striatum of normal adult rats cause the transient formation of large hemorrhagic cavities and do not survive. After differentiation by infection with Kirsten-ras murine sarcoma virus, and transplantation to the opposite striatum of the same brain, PC12 cells survive for at least 8 weeks and emit neurites. These neuron-like cells and their neurites retain tyrosine hydroxylase and choline acetyl transferase, as detected immunohistochemically. Thus, ras-primed PC12 cells may serve as a continuous source for both cholinergic and adrenergic transmitters, in vivo, without the need of exogenous nerve growth factor.