Ryosuke Murata

Sequential MRI, SPECT and PET in respiratory syncytial virus encephalitis.

Abstract We report on a 3-year-old girl with respiratory syncytial virus (RSV) encephalitis manifested by disturbance of consciousness, conjugate eye deviation, anuria, truncal ataxia and intention tremor. T2-weighted magnetic resonance imaging (MRI) showed hyperintense areas in the cerebellar cortex. No lesion was detected in the cerebral cortex, pons or spinal cord. The hyperintense areas in the cerebellar cortex diminished with recovery from the clinical manifestations and had resolved 2 months after onset. The MRI lesions in the cerebellum were considered to be due to oedema. SPECT and positron emission tomography (PET), performed 3 months after onset, disclosed areas of hypoperfusion and hypometabolism at the same sites. One year after onset, MRI showed mild atrophy of the cerebellum. Hypoperfusion on SPECT and hypometabolism on PET remained. Neuroimaging showed that ataxia and tremor in this case were the result of cerebellitis. The patient has no neurological deficit except for mild truncal ataxia. This patient is a rare example of RSV encephalitis.

Magnetic resonance images of tuberous sclerosis

SummaryThe cerebral lesions in tuberous sclerosis are of three kinds: subependymal nodules, cortical tubers, and cluster of heterotopic cells in the white matter. Understanding of these hamartomas is still incomplete even with modern imaging modalities. Magnetic resonance (MR) images of ten patients with tuberous sclerosis were reviewed and compared to computed tomographic (CT) scans and to the clinical severity of the disease. T2 weighted spin echo (TR=1800, TE=120) images and inversion recovery (TR=2100, TI=500–600, TE=40) images were obtained at the same axial planes. Periventricular nodules were better seen, because of their calcifications, with CT than with MR imaging. They were demonstrated as iso- to low intensity depending on the amount of calcification on T2 weighted images, and as a similar intensity to the white matter on IR images. Small peripheral lesions in the hemispheres, which were only occasionally seen as small low density areas on CT scans, were well demonstrated on MR images. These foci were hyperintense on T2 weighted images, and hypointense on IR images. Exact location of these was not in the cortex, but in the subcortical white matter. The findings indicate that these foci represent the pathologically well known demyelinating foci, which are commonly present under the cortical tuber, but may be independent of them. Cortical tubers were not confidently identified, which suggested that they might have similar intensity to the cortical gray matter. Some of the parenchymal calcifications other than periventricular nodules showed identical MR signal intensities to periventricular nodules, and the rest of the parenchymal calcifications had similar intensities to the subcortical lesions. This indicates that parenchymal calcifications can occur in the demyelinating white matter as well as in the heterotopic tubers in the white matter. The severely mentally retarded patients tended to have a higher number of subcortical lesions and no correlation was noted between the severity of mental retardation and either the number of periventricular nocules or ventricular dilatation.

Nucleotide sequences of the matrix protein gene of subacute sclerosing panencephalitis viruses compared with local contemporary isolates from patients with acute measles

Measles viruses isolated from brain cells of patients with subacute sclerosing panencephalitis (SSPE) have numerous mutations, especially in the matrix protein (M) gene. To find whether the M genes of these SSPE viruses were mutated randomly or in a pattern, we sequenced this gene from three strains of defective measles virus isolated in Osaka, Japan. We could deduce the sequence of the possible progenitor measles virus for each patient by comparison of the isolate with measles viruses prevailing at roughly the same time and place as the primary infection. Biased hypermutation affected the M genes of all three SSPE viruses, although the molecular mechanisms for the mutations might be various. Replacements of U with C in the plus strand accounted for 76% of all mutations in two of the strains, but in the other strain, replacements of A with G accounted for 52% of the mutations, and the U residues were unchanged.

Efficient isolation of subacute sclerosing panencephalitis virus from patient brains by reference to magnetic resonance and computed tomographic images

Subacute sclerosing panencephalitis virus has been isolated with difficulty from brains of infected patients. More strains are needed for the study of the pathogenesis of this virus. To make the isolation more efficient, we selected portions to be examined from the brains of three patients with reference to findings of repeated magnetic resonance and computed tomographic imaging. Three cell lines susceptible to measles virus field strains were used. In all three cases viruses were isolated most effectively from recent lesions and with Vero cells. Our results suggested that these imaging methods and Vero cells could be used for improvement in the efficiency of isolation of this virus from patient brains.

The matrix gene expression of subacute sclerosing panencephalitis (SSPE) virus (Osaka-1 strain): a comparison of two sibling viruses isolated from different lobes of an SSPE brain.

The Fr/V and Oc/V sibling viruses of the Osaka‐1 strain of the subacute sclerosing panencephalitis (SSPE) virus were defective in cell‐free virus production. By radioimmunoprecipitation assay, the matrix (M) protein was not detected in cells persistently infected with the Osaka‐1 strain. This undetectable expression was consistent with the selective reduction of antibody response to the M protein in the patient from whom the Osaka‐1 strain was isolated. The sequence of the M gene, however, predicted that the protein could be synthesized because the translational start and stop codons for the protein were not altered. Northern blot hybridization demonstrated the selective defect of the monocistronic mRNAs for the M protein and the phosphoprotein (P) together with the dominant presence of the P‐M bicistronic mRNA. This absence of the M mRNA was further confirmed by primer extension analysis. Therefore, the undetectable expression of the M protein in the infected cells was proved to be caused by a transcriptional defect. The two sibling viruses, isolated from remote portions of an SSPE brain, were indistinguishable in their viral characters, including the M gene sequences, which indicates the possibility of clonal expansion of the strain in the brain.

Developmental Study of Hippocampal Kindling

Summary: The critical susceptibility to seizure in immature hippocampus of rat brain was investigated using the kindling model of epilepsy. Hourly electrical low‐intensity stimulations applied to the left dorsal hippocampus of suckling rats induced longer initial AD duration (p < 0.01) with faster development of kindling seizure than in that of adult rats. In contrast to the marked reduction of wet dog shakes (WDS) in adult rats, in suckling rats persistent and prolonged WDS were observed as kindling progressed. The faster development of hippocampal kindling in suckling rats than in adult rats was explained by the different excitability of the stimulated brain site and the different extent of neural connections in many brain structures, which participate in generalization of seizures arising from a stimulated site.

Hemadsorption Expressed by Cloned H Genes from Subacute Sclerosing Panencephalitis (SSPE) Viruses and Their Possible Progenitor Measles Viruses Isolated in Osaka, Japan

Most subacute sclerosing panencephalitis (SSPE) viruses, including our Osaka‐1, −2, and −3 strains isolated in Osaka, have shown negative hemadsorption (HAD) by African green monkey red blood cells. This property has been thought to be characteristic of SSPE virus as compared to the positive reaction of the standard Edmonston strain of measles virus (MV). However, this assumption has become quite obscure because MV mutates frequently at the genetic level during its multiplication and also because recent field strains isolated by lymphoblastoid cell lines have shown negative HAD. To investigate the above issue, the nucleotide sequences of the hemagglutinin (H) genes from SSPE virus Osaka‐1, −2, or −3 strains were compared to those of various MV field strains isolated in Osaka by Vero cells. The H gene sequences of three SSPE strains were relatively conserved without such biased hypermutation as had been observed in the matrix (M) gene of three SSPE strains. However, this analysis of the H gene sequence of the SSPE viruses enabled us to deduce possible progenitor MVs, which are in agreement with the deduction from the M gene analysis we reported previously. The HAD of Vero cells transfected with the cloned H cDNAs from the SSPE strains and their progenitors suggested that negative HAD of the SSPE viruses has been maintained as one of original properties of the progenitor MVs rather than having been acquired as an altered one during long‐term persistent infection in the brains of patients with SSPE.

Different transcriptional expression of the matrix gene of the two sibling viruses of the subacute sclerosing panencephalitis virus (Osaka-2 strain) isolated from a biopsy specimen of patient brain.

Two sibling viruses of the subacute sclerosing panencephalitis (SSPE) virus Osaka-2 strain were isolated from a small biopsy specimen of the brain of an SSPE patient just before intraventricular interferon treatment by cocultivation with two different cell lines, Vero cells or B95a cells (Ogura et al, 1997). Both the virus-infected cells were found to be indistinguishable from each other in defective production of cell-free virus and in defective expression of the matrix (M) protein. The sequence analysis of the M genes predicted that they were translatable due to a lack of alteration of the translational start and stop codons for the proteins. A different pattern of the M monocistronic transcripts, however, was observed in a Northern blot analysis of the infected cells. This different pattern was confirmed further by a primer extension analysis. The undetectable expressions of the M proteins in the sibling virus-infected cells are most probably different in their molecular mechanisms. All these results indicate the possibility that the two different, replicable variants existed at Jabbour stage III of the diseases progression in a very small portion of the brain, where no lesion had yet been recognized by a magnetic resonance imaging.

Amygdala kindling and associated changes of entorhinal responses in suckling rats

Entorhinal field potential with amygdala stimulation in suckling (16-18 days old) and adult rats was recorded with a tungsten wire electrode (tip diameter 2-5 microns) to study the developmental changes in behavioral seizures and long-term potentiation (LTP) in the responses to amygdala kindling stimulations. Stimulating (twisted enamel-coated wires) and recording electrodes were implanted in anesthetized rats 2-3 days before kindling. The mean amplitude of the responses to test pulses (600 microA, 0.3 Hz) in the sucklings (0.58 mV) was smaller than in the adults (1.32 mV), and latency was about 3.3 ms longer. Kindling stimulations consisted of 0.5-ms monophasic rectangular pulses of 10 Hz with a 10-s train duration; the intensity was the afterdischarge (AD) threshold. Kindling stimulation in the sucklings usually increased the amplitude of the test responses evoked 10 min or 1 h after the kindling stimulation. The increased amplitude persisted for at least 24 h, showing LTP in the synaptic transmission. The LTP was especially prominent in the first kindling stimulation, and the LTP gradually increased with successive stimulations, with gradual progression of AD and the behavioral seizure stage as well. The mean number of kindling stimulations to cause generalized seizures in the suckling rats (10.5) was less than that for adults (12.5), and the continued evolution of LTP over the course of kindling was more or less easier in the sucklings than in the adults.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of hypoglycemia on kindling seizures in suckling rats

The effects of insulin-induced hypoglycemia on epileptic disorders of suckling rats were examined using an amygdala kindling model. Kindling stimulations were conducted at 16 and 17 days of age with electrodes implanted in the amygdala 2 days earlier. In 18-day-old kindled rats, which acquired generalized behavioral seizures (stages 4 and 5; Moshés score) by kindling stimulations, the duration of afterdischarge and behavioral seizures evoked by the stimulation at a threshold intensity to produce a generalized seizure was significantly prolonged after an injection of insulin (25 U/kg, i.p.). The prolongation was not observed in kindled rats that exhibited normal blood glucose concentrations after the application of saline or insulin together with glucose. There were no apparent changes in the amplitude of the afterdischarge, the score of behavioral seizure stages, or the generalized seizure threshold. A similar, marked prolongation of afterdischarge and behavioral seizures following the application of insulin, as in the kindled rats, was also observed during the course of the kindling acquisition without accelerating the development of kindling seizure scores. These results indicate that insulin-induced hypoglycemia easily increases the risk of prolonged seizures in immature brain without precipitating the secondarily generalizing mechanism.