Immunological and PCR analyses for Borna disease virus in psychiatric patients and blood donors in Japan.

The involvement of Borna disease virus (BDV) in psychiatric diseases in humans remains controversial. T-cell memory response and seroprevalence of BDV in patients with psychiatric disorders and blood donors in Japan were evaluated collectively by Western blot (WB) analysis with inhibition test, electrochemiluminescence immunoassay, immunofluorescence assay, and T-cell proliferative response as well as detection of BDV p24 RNA in peripheral blood mononuclear cells (PBMCs). Positive proliferative responses to both BDV p40 and p24 proteins were detected in 9% of patients with mood disorders (4 of 45), 4% of schizophrenic patients (2 of 45), and 2% of blood donors (1 of 45). By WB analysis, the antibody to BDV p40 was detected only in 2% of patients with mood disorders (1 of 45). The BDV p24 antibody was detected in 2% of patients with mood disorders (1 of 45) and 9% of schizophrenic patients. (4 of 45) No plasma reacted with both BDV proteins. The finding of a lower seroprevalence than previously reported suggests the presence of false-positive cases in the previous report. BDV RNA was detected only in 2% of patients with mood disorders (1 of 45). In these three serological assays, T-cell responses, and PCR analysis, there was no significant difference in the prevalence among the three groups. However, we found three psychiatric patients who were positive for both BDV antibodies and T-cell proliferative responses and one patient who was positive for BDV RNA in PBMCs. These findings suggest the usefulness of the proliferative T-cell response and that certain individuals are infected with BDV or a BDV-related virus.

The effects of N-methyl-D-aspartate (NMDA) and its competitive antagonist, 3-(2-carboxypiperazine-4-yl)propyl-1-phosphonic acid (CPP), injected into caudate-putamen on kindled amygdaloid seizures in rats.

N-methyl-D-aspartate (NMDA) is an agonist of NMDA receptors and 3-(2-carboxypiperazine-4-yl)propyl-1-phosphonic acid (CPP) is an NMDA receptor antagonist. NMDA (1 or 2 nmol per side) or CPP (2.5 or 10 nmol per side) was injected into the bilateral caudate-putamen of amygdaloid-kindled rats. In addition, CPP (10 nmol) was ipsilaterally or contralaterally injected into the unilateral caudate-putamen. Either 20 min after NMDA or 60 min after CPP, the kindled amygdala was stimulated at the generalized seizure triggering threshold. In a few animals tested, injection of NMDA into the bilateral caudate-putamen produced transient spiking activity, with no clinical manifestations. This feature began about 5 min after the injection and lasted about 10 s. When these animals were excluded from the statistical analysis, NMDA in the caudate-putamen showed a weak and non-significant anticonvulsant action. Injection of CPP into the bilateral caudate-putamen caused no ictal change, but markedly suppressed the kindled seizures. Injection of CPP into the unilateral caudate-putamen, regardless of the site, did not cause any ictal change, or affect the stimulation of the amygdala. These findings suggest that: (1) NMDA receptors in the caudate-putamen facilitate the development of kindled amygdaloid seizures; (2) activation of NMDA receptors in the bilateral, but not in the unilateral, caudate-putamen is required for the generalization and expression of kindled amygdaloid seizures.

Anticonvulsant action of metabotropic glutamate receptor agonists in kindled amygdala of rats.

1S,3R-1-Aminocyclopentane-1,3-dicarboxylate (1S,3R-ACPD) and L-2-amino-4-phosphonobutyrate (L-AP4) are selective agonists of metabotropic glutamate receptors (mGluRs). 1S,3R-ACPD (200 nmol) injected into the amygdala (AM) of non-kindled rats produced immediate and transient seizures, characterized by immobility, searching, and wet-dog shakes. However, this ictal response was not observed in AM-kindled rats. Intra-AM injection of 1S,3R-ACPD (40 or 200 nmol) or L-AP4 (200 nmol) resulted in a delayed and marked suppression of kindled seizures, 3 days after 40 nmol of 1S,3R-ACPD, 1 day after 200 nmol of 1S,3R-ACPD, and from 1 to 3 days after 200 nmol of L-AP4. These and the previous findings suggest that: (1) the excitatory action of 1S,3R-ACPD in the AM, which may be mediated by the postsynaptic mGluRs, is reduced after completion of kindling; (2) 1S,3R-ACPD and L-AP4 exert anti-convulsant action in the AM, presumably by activating the presynaptic as well as postsynaptic mGluRs.

Liposome-entrapped phenytoin locally suppresses amygdaloid epileptogenic focus created by db-cAMP/EDTA in rats.

Status epilepticus was induced in rats by injecting a combination of dibutyryl-cAMP (db-cAMP) and ethylenediaminetetraacetic acid (EDTA) into the amygdala (AM), and the effect of phenytoin (PHT), entrapped in liposomes (PHT-L) and given intravenously at 40 mg/kg, on the spiking activity of the AM epileptogenic focus was examined. Electroencephalograms were recorded from the db-cAMP/EDTA-injected AM and the bilateral sensorimotor cortices. One dose of PHT-L, given 30 min after intra-AM db-cAMP, produced immediate and transient seizure suppression, but did not suppress the sequential spiking activity, which lasted for more than 5 h. In contrast, two doses of PHT-L, given 30 and 60 min after intra-AM db-cAMP/EDTA, produced delayed and local suppression of AM discharges, and immediate and transient seizure suppression was also observed. The AM discharges began to be suppressed about 100 min after the second injection of PHT-L injection, with no overt change occurring in cortical spiking activity. This was followed by total seizure suppression about 170 min after the second PHT-L injection. This effect was not observed after one or two injections of PHT alone. When horseradish peroxidase (HRP), to which the blood-brain barrier is impermeable, was entrapped in liposomes (HRP-L) and given intravenously 30 min after intra-AM db-cAMP/EDTA, an accumulation of HRP was found in the db-cAMP/EDTA-injected AM in 2 of the 5 animals tested. With 2 doses of HRP-L given 30 and 60 min after intra-AM db-cAMP/EDTA, the local augmentation of HRP in the AM was found in all 5 of the 5 animals tested. Our findings suggest that: (1) the AM epileptogenic focus created by db-cAMP/EDTA has a high affinity for liposomes, and this factor participates in the local suppression of AM discharges by PHT-L, and (2) two injections of PHT-L are required for the AM to gather an effective amount of PHT-L.

Antiepileptic effects of inhibitors of nitric oxide synthase examined in pentylenetetrazol-induced seizures in rats.

The effects of intraperitoneal NG-methyl-L-arginine and N omega-nitro-L-arginine methyl ester, specific inhibitors of nitric oxide (NO) synthase, were examined on the pentylenetetrazol (PTZ)-induced seizures in rats. The incidence and latency for the onset of myoclonic jerks, clonic seizures, and tonic generalized extension were observed as specific parameters among PTZ-induced seizures. Both drugs preferentially suppressed the tonic generalized extension and prolonged the latency for the onset of each parameter, suggesting NO has a significant effect on the PTZ-induced seizure.

The effect of thyrotropin-releasing hormone (TRH) on limbic status epilepticus in rats.

We produced limbic status epilepticus in rats by injecting a combination of dibutyryl-cAMP (db-cAMP) and ethylenediaminetetraacetic acid (EDTA) into the amygdala (AM). Thirty minutes after intra-AM db-cAMP/EDTA injection, thyrotropin-releasing hormone (TRH) was administered intravenously or intracerebroventricularly. Intravenous TRH (3, 25, 50 mg/kg) produced immediate activation of electroclinical seizures, lasting for 25-45 min. In some animals which showed this seizure activation, complete seizure suppression occurred 55-70 min after the TRH treatment. Similar activation of ictal seizures with delayed seizure suppression was obtained after intracerebroventricular TRH (25, 50 micrograms). The findings suggest that the effects of intravenous TRH are due to its central action and that the use of intravenous TRH is not a promising approach for the treatment of status epilepticus.

Kindled amygdaloid seizures in rats cause immediate and transient increase in protein kinase C activity followed by transient suppression of the activity.

Protein kinase C (PKC) activity in hippocampus and amygdala was measured during kindled seizures and 30 min, 3, 24, and 48 h, and 2 weeks after seizures in amygdaloid-kindled rats. Sham-operated rats not subjected to kindling were used as controls. During kindled seizures, membrane-bound PKC activity in bilateral hippocampi was significantly increased, with a slight reduction in cytosolic PKC activity, but there was no change in either membrane-bound or cytosolic PKC activity in bilateral amygdala. Thirty minutes after seizures, PKC activity in both fractions was significantly increased in bilateral hippocampi and amygdala. Three hours after seizures, PKC activity in both fractions was markedly decreased in bilateral hippocampi. In bilateral amygdala, a similar and significant decrease in membrane-bound PKC activity was noted, with no marked change in the cytosolic fraction. Twenty-four hours after seizures, a significant decrease in membrane-bound PKC activity in bilateral hippocampi and amygdala was again noted, although cytosolic PKC activity was unchanged. Forty-eight hours after the seizures, PKC activity in both fractions had returned to control levels. Two weeks after the last seizure, there was no significant change in PKC activity in either fraction in any region, except for a slight increase in membrane-bound PKC activity in unilateral hippocampus contralateral to the kindled amygdala. These results suggest that kindled amygdaloid seizures cause an immediate and transient increase in PKC activity in limbic structures, followed by suppression of enzyme activity, and that PKC in hippocampus responds to kindled seizures more readily and preferentially than it does in amygdala.(ABSTRACT TRUNCATED AT 250 WORDS)

Anticonvulsant effect of liposome-entrapped superoxide dismutase in amygdaloid-kindled rats.

Amygdaloid-kindled rats received intravenous human copper-zinc superoxide dismutase (CuZn-SOD) either in free form or entrapped within liposomes (SOD-L), at 5, 10 or 20 mg/kg. The animals were stimulated at the generalized seizure-triggering threshold 5 min, 2 h and then every 24 h after the drug was given, until 5 consecutive stage 5 seizures were induced. Free CuZn-SOD had little or no effect. However, SOD-L, particularly at 10 mg/kg, had a prolonged anticonvulsant effect, although there was great individual variation in the onset and duration of seizure suppression. This effect of SOD-L may be due to the ability of liposomes to act as a depot for the sustained release of drugs.

An important role of non-NMDA receptors in the expression of kindled amygdaloid seizure in rats.

gamma-D-glutamylaminomethylsulphonic acid (GAMS), a preferential antagonist of non-NMDA receptors (kainate and quisqualate receptors), was injected into the kindled amygdala (AM) of rats. When the kindled AM was stimulated at the previously established generalized seizure triggering threshold (GST) one hour after the GAMS (1 or 2 mumol) injection, afterdischarge (AD) generation was completely suppressed. However, a re-stimulation at the intensity of 40-200 microA above the GST generated AD associated with Stage 1 or 5 seizure. Our result suggests an important role played by non-NMDA receptors in the expression and generalization of AM-onset seizures.

[Effects of thyrotropin-releasing hormone (TRH) on status epilepticus in rats].

Recent studies have demonstrated that intramuscular administration of thyrotropin-releasing hormone (TRH) or its analogue improves various clinical aspects of intractable epilepsy such as Lennox-Gastaut syndrome, West syndrome, and myoclonus epilepsy. Other clinical studies reported efficient property of intravenous TRH against status epilepticus. However, it is also true that intravenous TRH produces epileptic seizures in patients with epilepsy or organic brain damage. Thus, the utility of intravenous TRH for the treatment of status epilepticus seems to be equivocal. To further explore the problem in this regard, we examined the effect of TRH on limbic status epilepticus in rats. Thirty-eight male Wistar rats weighing 180-220g were used. Status epilepticus was induced by intracerebral injection of a combination of 200 micrograms of dibutyryl-cAMP (db-cAMP) and 67.2ng of ethylenediaminetetraacetic acid (EDTA) into the amygdala (AM) through an implanted cannula. 30 min later, TRH or vehicle (distilled water) was administered intravenously (i.v.) or intracerebroventricularly (i.c.v.). Although 3 mg/kg of TRH (n = 9), when injected i.v., did not alter the pattern of electroclinical ictal responses induced by db-cAMP/EDTA, 25 mg/kg (n = 5) and 50 mg/kg (n = 5) of TRH significantly exaggerated EEG and/or behavioral ictal seizures, beginning immediately after the injection and lasting for more than 30 min. With 50 mg/kg of TRH, the exaggerated seizure patterns were followed by marked suppression of electroclinical seizures. 50 micrograms of i.c.v. TRH (n = 5), like higher doses of i.v. TRH, caused a slight, but not a significant, build up of electroclinical ictal seizures, beginning immediately after the injection and lasting for about 30 min.(ABSTRACT TRUNCATED AT 250 WORDS)