Novel acyl-CoA synthetase in adrenoleukodystrophy target tissues.

X-linked adrenoleukodystrophy (X-ALD) is a neurodegenerative disorder characterized by demyelination of white matter. The X-ALD gene product adrenoleukodystrophy protein (ALDP) is expressed broadly among various tissues. However, deficiency of functional ALDP exclusively impairs brain, adrenal gland, and testis. Thus, loss of ALDP function is assumed to involve inactivation of a putative mediating factor that functions in a tissue-specific manner. Here we cloned a mouse cDNA encoding a novel protein, Lipidosin, that possesses long-chain acyl-CoA synthetase (LCAS) activity. Lipidosin is expressed exclusively in mouse brain, adrenal gland, and testis, which are affected by X-ALD. LCAS activity of Lipidosin was diminished by mutation of conserved amino acids within the AMP-binding domain. Mutation of the Drosophila homologue of Lipidosin has been reported to cause neuronal degeneration. Thus, Lipidosin may mediate the link between ALDP dysfunction and the impairment of fatty acid metabolism in X-ALD.

Isolation of a differentiation-defective myoblastic cell line, INC-2, expressing muscle LIM protein under differentiation-inducing conditions.

A non-differentiating myoblastic cell line, INC2, and a differentiating cell line, COM3, were established from the mouse myoblastic cell line C2C12. Under differentiation conditions, both COM3 and INC2 cells stopped proliferation in a similar manner. The COM3 cells then differentiated into myotubes during the 4-day differentiation culture. In contrast, almost none of the INC2 cells differentiated into myotubes even in differentiation medium. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblot analyses showed that the levels of myogenin and MyoD proteins were significantly decreased in INC2 cells. The differentiation marker sarcomeric myosin heavy chain (MHC) was expressed in COM3 but not in INC2 cells. In contrast, both INC2 and COM3 cells expressed another myogenic regulatory factor, muscle LIM protein (MLP), in a differentiation condition-dependent manner. These results suggest that MLP gene expression is regulated in a myogenin/MyoD-independent manner. Enforced expression of the myogenin gene induced MHC expression in INC2 cells. Thus, the signaling pathway situated downstream is assumed to be intact in INC2 cells and suppression of myogenin, gene expression may be a primary defect in INC2 cells.

Role of tyrosine kinase in the regulation of myogenin expression.

Using an affinity-purified anti-myogenin antibody, three stages of mouse myoblast C2C12 cells during myogenesis could be identified: proliferating myoblasts as myogenin-negative mononucleated cells, differentiating myoblasts as myogenin-positive mononucleated cells, and myotubes as myogenin-positive multinucleated cells. We found differential effects of genistein, an inhibitor of protein-tyrosine kinase, on myogenic cells during these three stages. Genistein severely inhibited myotube formation and myogenin production in differentiating myoblasts by inhibiting the transcription of the myogenin gene in a dose-dependent manner. We also found that genistein inactivated mitogen-activated protein kinase (MAP kinase) accompanied by suppression of myogenin expression. In contrast, genistein failed to inactivate MAP kinase and eliminate myogenin from myotubes. The results suggest that protein-tyrosine kinase plays a role in the transcriptional regulation of myogenin through the MAP kinase cascade during myogenesis. Furthermore, genistein inhibited the transactivation of the myosin heavy chain gene by constitutively expressed myogenin. Therefore, it is suggested that protein-tyrosine kinase is involved in the post-translational regulation of myogenin as well as in transcriptional regulation during myogenesis.

Phosphorylation of a proline-directed kinase motif is responsible for structural changes in myogenin.

Myogenin, a member of the MyoD family which governs skeletal muscle differentiation, was identified as a pair of phosphorylated bands on SDS-PAGE during myogenesis. The slow migrating form was found to be hyperphosphorylated myogenin. In vitro phosphorylation by CDC2 kinase caused a prominent reduction in electrophoretic mobility of myogenin. Furthermore, we demonstrated that phosphorylation of the serine residue at position 43 contributes to the modification of myogenin in vivo and in vitro resulting in the reduction in electrophoretic mobility. We propose here that a CDC2-like proline-directed kinase regulates myogenin activity through its phosphorylation.

[Cerebral hemorrhage in a case of antiphospholipid antibody syndrome].

We describe a 36-year-old man with antiphospholipid antibody syndrome complicated by cerebral hemorrhage. In December 1991 he was brought to another hospital with sudden onset of left hemiparesis and status epilepticus. He had been well previously. A CT scan and MRI showed a cerebral hematoma located in the right frontoparietal region. Twelve days later he was transferred to our hospital. Although a CT scan, MRI, and cerebral angiography were repeated, they did not reveal any abnormality regarding an etiology. Only persistently abnormal finding in laboratory studies was positive for lupus anticoagulant and anticardiolipin antibody, i.e. antiphospholipid antibodies. There was no serological evidence of SLE or other autoimmune diseases. Stereotactic biopsy of the hematoma wall and scalp artery showed no abnormality. Based on above findings we conclude that antiphospholipid antibodies have played an important role for the hemorrhage. Antiphospholipid antibody syndrome should be considered in a case of an unexplained cerebral hemorrhage especially in a young and normotensive patient.

Referential derivation of epidural electroencephalogram in El mice.

Epidural EEG of 20 El mice is recorded under ketamine and xylazine anesthesia with epoxy coated silver balls as exploring electrodes. A stainless steel needle is placed subcutaneously near the nose as a reference electrode. Reproducibility of recording is remarkable. Multiple spike complex synchronizing on both frontal lobes is observed in 8 mice. Among them, small spikes preceding the multiple spike complex are also observed on the left frontal lead in 7 mice and on the right in 1 mouse. These spikes do not spread to occipital recordings. The discriminative ability of the recording is due to the coated electrode commonly employed with much larger animals and to suitable placement of the reference electrode. This simple and well known method permits the analysis of not yet satisfactorily explored the EEG of El mouse.

[Clinico-histological study of low-density non-enhancing glioma].

Some gliomas are not noticeably enhanced on initial computer tomography (CT) scans. Such low-density non-enhancing gliomas have a relatively long period between onset and surgery ranging from 5 to 25 months. The mechanism causing the low density over a certain period of time in the sequential CT findings was retrospectively investigated. Characteristic histological findings associated with such low-density areas were microcystic formation, necrosis, intratumoral edema, infiltrative growth, and absence of capillary proliferation. Proliferation of capillary vessels is characteristic of CT enhancement.

[Magnetoencephalographic localization of epileptic foci using a 37-channel biomagnetometer].

The magnetoencephalography (MEG) and electroencephalography (EEG) were recorded simultaneously from 10 normal subjects using a 37-channel biomagnetometer. No paroxysmal spikelike waveform was observed in MEG at rest with eyes closed. The MEG and EEG were recorded also from 16 patients with primary epilepsy and 24 patients with secondary epilepsy. The examination proved to be safe for both normal subjects and patients with epilepsy. Interictal spikes were observed in 27 cases during the examination. The percentage of spikes identified in MEG but not in EEG was found to be 2. 3% of all spikes. The foci of the spikes identified in MEG were localized and determined in 20 cases. In 10 patients with secondary epilepsy, the localization of the foci were compared with the lesion demonstrated by magnetic resonance imaging (MRI) or computerized tomography (CT) and with the findings of EEG. In 6 cases, the foci by MEG were consistent. In the 4 cases where the MEG foci did not correspond to the MRI or CT findings, MEG foci were supported by the findings of EEG. MEG allows three-dimensional localization and enables us to elucidate the propagation of paroxysms. MEG was very useful in diagnosing epilepsy.

[An autopsy case of glioblastoma occurred in the region after lobotomy].

The authors describe an autopsy case of glioblastoma occurred after 38 years received lobotomy. The patient was a 72 year-old male, who received lobotomy at 34 year old against schizophrenia. CT scan taken at 72 year old showed irregular low density areas without mass effect in the bilateral frontal white matter adjacent to the anterior horn. After 4 months, the signs of intracranial hypertension were observed and his consciousness was disturbed abruptly. CT scan revealed ring enhancement with marked mass effect in the left frontal lobe. A biopsy specimen from the tumor showed a picture of anaplastic astrocytoma. Family rejected the remission maintenance treatment. The patient died 3 months later the onset. At autopsy, a large tumor occupied in the left frontal lobe was recognized. The tumor demarcated poorly from the cerebral tissue and invaded into the left anterior cingulate gyrus and the corpus callosum. Histologically, tumor cells composed of fibrillary, gemistocytic and multinucleated astrocytes. GFAP, NSE and vimentin were found in large cells. Histological diagnosis was glioblastoma. It was suggested that the tumor occurred from the region around a cyst of prefrontal lobotomy in the left frontal lobe. In the right frontal lobe, a large cyst in size of 30-18 mm was present in the centrum semiovale. The wall of cyst was composed of layer of glial scar tissue. The origin of the cyst was discussed.

[Cytochemical study of Mg2(+)-ATPase and ALPase activity in human meningiomas].

Ultracytochemical features of microvessels and tumor cells of the human meningiomas were examined by light and electron microscopy with special reference to the distribution of Mg2(+)-ATPase and alkaline phosphatase (ALPase) activity on the walls of the vessels and tumor cell surfaces. Materials used were 4 cases of meningiomas, 2 of which were meningotheliomatous type, one fibroblastic type and one malignant meningioma respectively. For ultracytochemistry, specimens were quickly fixed in an ice-cold 0.1 M cacodylate buffer containing 8% sucrose (pH 7.2) for one hour and transferred to a substrate solution for detection of Mg2(+)-ATPase and ALPase. The preparations were incubated at 37 degrees C for 15-30 min in the medium described by Mayahara et al. for ALPase and for 15-20 min in the medium described by Wachstein and Meisel. The control samples were incubated in a medium containing 1 mM Bromotetramisole for ALPase and also in a substrate free medium for Mg2(+)-ATPase. At the light microscopy, Mg2(+)-ATPase and ALPase activities appeared to be mainly restricted to the capillary wall and around or in the tumor cell nest showing whorl formation. Both enzyme activities were negative in the control study. By electron microscopy, reaction products representing Mg2(+)-ATPase activity were distributed in the basolateral plasma membrane of the endothelial cells on the surface of the pericytes and on the surface of the tumor cells. Reaction products of ALPase activity located mainly on the abluminal surfaces of the endothelial cells and in some specimen on both luminal and abluminal surfaces of those cells. Intense reaction products were distributed evenly on all round surfaces of the tumor cells.(ABSTRACT TRUNCATED AT 250 WORDS)

[Clinicopathological study on low grade glioma. In relation to malignant transformation].

The clinical status of patients with glioma is influenced by 1) the histological malignancy of the tumor, 2) the tumor volume, 3) secondary status such as brain edema or intracranial hypertension due to the tumor, and 4) the host immunity. Due to some improvement in at least 2) and 3) by the initial treatment, most low grade glioma cases pass through a clinically silent postsurgical period. However, at a certain point, transition to a high grade tumor malignant transformation may occur with exacerbation of the symptoms. Twenty-two cases of histologically established low grade glioma experienced over the past 7 years, in which immunological status was evaluated, were analyzed. Nine cases (41%) showed malignant transformation. Characteristic pictures of the clinical symptoms, computed tomography (CT) scan findings, immunological status, and morphological findings (mainly immunohistochemical examination) in nine cases were delineated. The findings at the time of exacerbation of the symptoms were as follows. In all cases CT scan demonstrated the change in the main lesion from low density to mixed density and were compatible with a high grade glioma. Reduction in host immunity was verified. Morphological increase in the tumor volume, increase in histological malignancy and deterioration in the secondary status due to the tumor were confirmed. Necrosis of the tumor cells as well as increase in giant cells and gemistocytes were observed. Immunohistochemical analysis revealed a decrease and irregularity in glial fibrillary acidic protein positive cells and positive processes as well as increase in vimentin intensity. These findings demonstrate change in the biological characteristics of the tumor.(ABSTRACT TRUNCATED AT 250 WORDS)

[Brain metastasis from primary pericardial mesothelioma. Case report].

A 50-year-old female was admitted because of nausea, vomiting, and cerebellar ataxia. Computed tomography scan revealed an enhanced mass accompanied with a cyst in the right cerebellar hemisphere. The mass situated in the subcortical region was removed. Histologically, highly vascular tumor cells lined the cavities. Postoperative radio- and chemotherapy were administered and the clinical symptoms improved gradually. Two months later, the patient complained of dyspnea. Chest X-ray on second admission demonstrated cardiomegaly. Hemorrhagic pericardial effusion amounting to 1000 ml was aspirated by pericardial puncture. Papillary clusters of tumor cells were demonstrated in the pericardial effusion. The patient died of cardiac failure. At necropsy solid tumors were located in the heart, lung, left inguinal region, and cerebellum. Histological diagnosis was mesothelioma arising from the heart. Primary pericardial mesotheliomas are rare; approximately 106 cases have been reported. Pericardial mesothelioma frequently spreads to the adjacent pleura and mediastinum, but distant metastases are extremely rare because patients with pericardial mesothelioma tend to die early due to cardiac failure or cardiac tamponade.

[Morphologic findings and biologic behavior in the high grade glioma--a postmortem study of 22 cases].

Morphologic features of the autopsied specimen of 22 cases with supratentorial gliomas treated by surgery, radiation and/or chemotherapy were analysed, and the characteristics of recurrence of gliomas were searched for. The cases consisted of anaplastic 12 astrocytoma and 10 glioblastoma. The results were as follows: 1) Characteristic CT findings before death were regrowth of the tumor mass or the occurrence of a new enhanced lesion in 21 out of 22 cases. The enhanced lesion showing regrowth of the tumor located in the same site as the previous tumor mass in 21 cases. The new enhanced lesion resulting from a trans-or subependymal tumor spread, was seen in the ventricular wall, and these findings were a characteristic feature of the recurrence of gliomas. 2) Modes of extension of the tumor were subdivided into 3 types. One was the expansive or infiltrative type caused by regrowth of the residual tumor. In the second pattern, a spread of tumor cells occurred along the myelinated fiber tracts to the brain stem (60%), or to the contralateral cerebral hemisphere through the corpus callosum (50%). The third mode of tumor propagation was cerebrospinal fluid seeding with intraventricular or subarachnoid tumor regrowth (45%). 3) Characteristic histological findings shown in the original tumor bed were those of increased cellularity with endothelial proliferation, widespread necrosis with occlusion of the blood vessels, occurrence of the gemistocytic astrocytes and large bizarre cells. Thickening of wall of the blood vessels due to effect by radiation was followed by occlusion of the blood vessels. Large necrosis in the tumor tissue was caused by those process and others. Necrotic area was mainly circumscribed and corresponded to the territory of the vessels. One of the specific findings in the morphological changes of the tumor cells was giant cell formation which were monstrous cell, giant cell (12 cases out of 22), and gemistocytic cell (in all cases). These specific cells were supposed to the degenerative changes of the tumor cells exposed while withstanding such adverse conditions as hypoxia, radiation and chemotherapy. 4) Infiltration distant from the primary lesion which were defined only by microscopical examination was demonstrated as both through myelinated fiber tracts in 8 cases and through perivascular spaces in 2 cases. Reinvasion of the tumor cells from the subarachnoid spaces to the brain parenchyma was along the Virchow-Robins spaces of the penetrating blood vessels in the latter cases.(ABSTRACT TRUNCATED AT 400 WORDS)

[Infected chronic subdural hematoma due to an ethmoiditis; a case report].

The patient was a 50-year-old female, and the initial symptoms were disturbance of consciousness and fever. CT scan showed a chronic subdural hematoma and left ethmoiditis. Emergency surgery disclosed infected chronic subdural hematoma. We classified this as infected chronic subdural hematoma caused by ethmoiditis.

Penetration of imipenem and other antibiotics into inflammatory exudate and their efficacy in pseudomonas infection in the rat granuloma pouch model.

The rat granuloma pouch model was used to study the penetration of imipenem into inflammatory exudate and its efficacy in an experimental local infection. Rats were given a single intravenous injection of drug via the tail vein at a dose of 40 mg/kg. The peak concentrations of imipenem, ceftizoxime, cefazolin, and cefsulodin in exudates ranged from 10.1 to 12.3 mg/l, except that ampicillin achieved only 5.4 mg/l. Imipenem rapidly reached peak concentration after injection, but the half life for elimination of imipenem from exudate was 77.7 min, which was shorter than that of other beta-lactam antibiotics. Imipenem exhibited more rapid reduction in bacterial growth than cefsulodin in an experimental local infection with Pseudomonas aeruginosa in the pouch model. The results suggested that imipenem was an effective agent in treating the local infection.