Well maintained expression of CYP genes in sandwich-culturing hepatocytes: quantitative analysis using real-time PCR method.

Sandwich-culturing is an excellent hepatocyte culturing method in drug metabolism studies, however, its advantages for gene expression of cytochrome P450 (CYP) have not been evaluated so far. The present study was undertaken to determine the utilities of sandwich-culturing hepatocytes for evaluation of CYP genes expression. Hepatocytes from male rats were cultured for 5 days between two layers of type-I collagen gel (sandwich-culturing) or over type-I collagen gel (single gel culturing). To determine the expression of CYP genes rapidly and accurately, the time course study using real-time RT-PCR quantification was conducted in the present study, and CYP2B1, CYP2B2, CYP3A2, CYP3A9 and CYP3A23 genes were measured. Albumin secretion was also measured by ELISA to evaluate cell viability. Higher expression and excellent maintenance of all CYP genes were confirmed in sandwich-culturing hepatocytes than those in single gel culturing. Particularly, significant difference in the amounts of CYP genes expression was observed between both methods after 3 days culturing. Albumin secretion was also higher in sandwich-culturing after 3 days culturing, suggesting that the cell viability of hepatocytes was maintained. These results indicate that sandwich-culturing method is much more advantageous than the ordinate method in maintaining the CYP gene expression and cell viability.

Ultrahigh-vacuum reaction apparatus to study synchrotron-radiation-stimulated processes.

An ultrahigh-vacuum reaction apparatus to study synchrotron-radiation-stimulated processes has been constructed and placed on beamline 4B of the synchrotron radiation storage ring (UVSOR) at the Institute for Molecular Science. The apparatus is designed so that multiple synchrotron radiation processes such as etching and chemical vapour deposition can be carried out successively without breaking the high vacuum. It is equipped with IR reflection absorption spectroscopy (IRRAS) apparatus and reflective high-energy electron diffraction (RHEED) apparatus for in situ observations. The basic parameters of the apparatus including etching and deposition rates have been measured. IRRAS using buried metal layer substrates has been confirmed to be a very useful method of analyzing the reaction mechanisms of the synchrotron-radiation-stimulated processes.

[Traumatic hematoma in the basal ganglia (caudate) with favorable prognosis: report of two cases].

Two cases of traumatic basal ganglial hematoma with fairly good prognosis were reported. Several cases with similarly favorable prognosis could be also found in the recent literature. In these cases, post-traumatic disturbance in consciousness was mostly slight or moderate and the patients were rarely comatose. The inquiries in these cases were commonly associated with superficial injuries such as skull fracture, epi-or sub-dural hematoma, brain contusion, or another traumatic ICH. The hematomas in the basal ganglia usually showed a mass effect causing focal neurological signs such as hemiparesis. Neurological improvement was achieved in the operated cases, and final outcome was mostly fair with some fully recovered cases. These clinical features are quite different from those of most cases of traumatic basal ganglial hematoma reported so far, which closely resemble diffuse axonal injury and whose prognoses are extremely poor. There may be two different categories in the traumatic basal ganglial hematomas, those with fair outcomes, and those with poor outcomes.

[Coexistence of intracranial and spinal cavernous angiomas: case report].

A case of a 43-year-old man with coexistence of intracranial and spinal cavernous angiomas is presented. The patient had a 2-year history of severe back pain incurred by neck flexion, and he became aware of weakness of the right lower extremity and paresthesia of the left lower extremity. Neurological examinations at the time of the first admission demonstrated incomplete Brown-Séquard syndrome. Myelograph, myelo-CT and contrast enhanced CT showed an intramedullary mass at the Th3-Th5 level. The patient received laminectomy with total removal of the lesion. Pathological diagnosis was cavernous angioma. Six years later, the patient complained of subacute weakness and numbness of the left upper extremity. Head CT demonstrated a high density lesion of about 2cm in diameter in the right frontal lobe. MRI showed a mixed signal intensity lesion with a marked low-intensity rim in the same area. Total extirpation of the lesion was performed. Pathological diagnosis of the intracerebral lesion was also cavernous angioma. Intramedullary cavernous angioma is very rare. Furthermore, bifocal cavernous angiomas involving both the spinal cord and the brain are extremely rare, and, only 5 cases have been reported in the literature. To our knowledge, this is the first case diagnosed by surgical specimens of coexisting intramedullary and intracerebral lesions.

Changes of plasma thromboxane level in subarachnoid haemorrhage. A study with 11-dehydro-TXB2 as measuring index.

Changes of plasma thromboxane level in subarachnoid haemorrhage (SAH) were studied clinically and experimentally using 11-dehydro-thromboxane B2 (11 DTX) as a measuring index. 11 DTX is a major long-lived metabolite formed from thromboxane (TX) B2, and is said to be a more reliable parameter for detecting TXA2 production in biological systems. In this clinical study, blood was sampled from the cubital vein of 10 SAH patients on the earliest possible day (day 0 or 1), during the vasospasm predilection period (day 7 approximately 11) and in the chronic stage (day 16 approximately 32). Plasma concentrations of 11 DTX and 6-keto-PGF 1 alpha were measured in clinical cases. A canine SAH model was produced by the two haemorrhage methods and blood was sampled from the superior sagittal sinus before and on day 4 of the first cisternal blood injection. 11 DTX, TXB2 and platelet function were examined in each sample. In the clinical studies, plasma 11 DTX levels tended to be higher in the early stage of SAH but decreased thereafter to the normal or lower level. Plasma concentrations of 6-keto-PGF1 alpha tended to decrease mildly during the vasospasm predilection period. In the experimental study, neither definite change of plasma 11 DTX level nor neurological deficit could be induced by the mimic SAH, while an increase in platelet aggregability and narrowing of the basilar artery were observed. 11 DTX was inferred to be a more reliable parameter of TX biosynthesis than TXB2.

[Cyst formation of meningioma: report of two cases and review of literature].

Cystic meningioma is rare and said to account for 1 to 2% of all types of meningioma. Here we report two cases of cystic meningioma. Case 1 is a 25-year-old man. He had suffered from sporadic epileptic seizure for about two years until the first CT was performed. It revealed a large cystic lesion at the right parietal region in touch with the midline structure. A nodular high density area was delineated in the contrast enhanced CT on the medial wall of the cyst. He showed no neurological abnormality at all. Case 2 is a 56-year-old female who had been complaining of memory disturbance, and an episode of sudden dizziness followed by a syncope that lasted for a few minutes. Neurological examination revealed slight right hemiparesis, right homonymous hemianopsia and left papilledema. A large cystic lesion was delineated at the temporal region in the plain CT, and no positively enhanced region could be found in the contrast enhanced CT. After surgical removal of the tumors, both cases were identified as cases of meningioma. They were diagnosed histopathologically as a fibroblastic meningioma and a meningotheliomatous meningioma respectively. The wall of the cyst consisted of the cortical surface per se in both cases. Preoperative diagnosis of such cases and the mechanism by which the cyst is formed were also discussed.

[Disturbance of inhibitory capacity of endothelial cell for platelet adhesion or aggregation following experimental subarachnoid hemorrhage].

Time sequential changes of the endothelial cells of feline basilar arteries after experimental subarachnoid hemorrhage (SAH) were studied morphologically and functionally under the scanning electron microscope (SEM). Experimental SAH was induced by the two-hemorrhage method, and the basilar artery was removed at 1 hour, or 2, 4, 7, or 14 days respectively after the 1st cisternal blood injection. At each stage, morphological changes of the luminal surface and endothelial cells of the basilar artery were observed by SEM. In another group, adenosin diphosphate (ADP) was infused through the right vertebral artery at the various stages mentioned above, for the purpose of activating platelets, and the basilar artery was then removed. The platelet adhesion or aggregation on the luminal surface was observed quantitatively also by SEM, for evaluating the inhibitory capacity of the endothelial cells upon platelet adhesion or aggregation after SAH. The fold formation of the arterial luminal surface and the ballooning of the endothelial cells were observed initially at 1 hour and 2 days respectively after the 1st cisternal blood injection. The latter findings became most remarkable during the 4 to 7 days after the said blood injection. They were accompanied by intercellular gap formation. None of these findings were seen, however, in the specimens taken 14 days after the said blood injection. In the group with pre-sacrificing ADP infusion, adhesion or aggregation of platelets on the arterial luminal surface was scarcely seen before the second day, but became remarkable 4 to 7 days after the 1st cisternal blood injection, and were reduced by the time 14 days had passed.(ABSTRACT TRUNCATED AT 250 WORDS)

[Subependymoma of the septum pellucidum. MRI features and its usefulness in planning surgery].

A 61-year-old female was referred because of headache and gait disturbance. A computed tomography (CT) scan revealed a midline isodense mass lesion. The limit of the tumor was equivocal even after administration of contrast medium. Magnetic resonance imaging (MRI) study, however, clearly showed the relationship between the tumor and the surrounding structures, such as the corpus callosum, the ventricular cavities. The tumor had originated from the region of the septum pellucidum and positioned just beneath the corpus callosum from the genu to the splenium with the roof of the third ventricle pushed downward. The tumor was totally extirpated via the interhemispheric paratranscallosal route. The tumor was typical subependymoma. The post-operative course was uneventful. Transient psychiatric disturbance and inability to retain recent memory were observed and the former subsided in several weeks. CT and MRI characteristics of subependymoma are summarized and the usefulness of MRI to locate tumors and to plan proper surgical approaches are emphasized.