Human standard astroviruses, serotypes 1 to 7, and 35 Japanese isolates were typed by reverse transcription and polymerase chain reaction (RT-PCR) with serotype-specific primers for the first time. The results were identical with those obtained by enzyme immunoassay with serotype-specific polyclonal antibodies, a method which has already been reported. RT-PCR with serotype-specific primers is useful for epidemiological studies of astroviruses where serotype-specific polyclonal antibodies are not available. Two parts of the capsid region, N terminus and C terminus, were sequenced. Serotypes differed in those regions. The N terminus differed less than the C terminus between serotypes. Both the N terminus and C terminus were similar intraserotypically with the exception of serotype-4 isolates which could be divided into A and B subgroups on the basis of their C terminus sequences, which were not known previously. Human astroviruses are non-enveloped, single-stranded RNA viruses which were first identified in 1975 by the electron microscopy of stool specimens from children with diarrhea (1, 8). Not only sporadic cases but also outbreaks of astroviruses have been reported (9,13,19). Astroviruses can be cultured in LLC-MK2 cells and CaCo-2 cells (5,22). From the cultured viruses, seven human astrovirus serotypes have been identified serologically by using serotype-specific polyclonal antibodies (6). Serotype 8 was recently reported in a database (Accession No. Z66541).The complete genomic sequences of human astrovirus 1 were determined in 1994 (7, 23). After that, the total capsid sequences of serotypes 2, 4 and 6 were reported (3, 6, 11,24). The partial sequence of the capsid region of serotype 3 is also known (12). These results enabled us to detect and sequence astroviruses by reverse transcription and polymerase chain reaction (RT-PCR) (16).We obtained and cultured the standard astroviruses, serotypes 1 to 7. We then analyzed the full length of the capsid regions by RT-PCR and sequencing (preparing for submission). From these results, we designed serotypespecific oligonucleotide primers in the capsid region. We then compared the serotypes as determined by RT-PCR and enzyme immunoassay (EIA) using serotypespecific polyclonal antibodies on Japanese strains of astroviruses. Moreover, we compared the nucleotides and the predicted amino acid sequence homologies in two parts of the capsid region.Our main purpose was to verify that RT-PCR with serotype-specific primers is applicable to the epidemiological study of astrovirus serotypes as compared to serotyping by EIA and sequence analysis because serotype-specific antibodies are commercially unavailable and there are limited epidemiological studies. In addition, the sequence of the capsid region is analyzed in two parts of the relatively conservative portion, N terminus, and relatively variable portion, C terminus, to deter-

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We studied the clinical efficacy of allopurinol as add-on therapy in 31 patients with intractable epilepsy. When administered for a short time, allopurinol was effective in 17 patients (55%); 8 were seizure-free, 8 had 75% decrease in seizure frequency, and 1 had greater than 50% decrease. Allopurinol was most effective in patients with localization-related epilepsy, especially in secondarily generalized tonic-clonic seizures. Allopurinol was not as effective in patients with Lennox syndrome or West syndrome, or in severe myoclonic epilepsy in infants. When allopurinol was administered greater than 1 year, its initial effectiveness continued in 8 of 14 patients who exhibited initial improvement. In 2 of the remaining 6 patients, the initial improvement disappeared during the course of treatment but control was regained by increasing the dosage of allopurinol. Mild side effects were observed in 4 patients (13%): drowsiness in 3 and abdominal pain in 1. Allopurinol may be a useful antiepileptic drug (AED), and a double-blind placebo-controlled trial should be performed.

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The entire capsid regions of 12 serotype-4 astroviruses from Japan were sequenced and compared with those of other serotypes. Serotype-4 isolates were divided into two new subgroups. The intrasubgroup nucleic acid and deduced amino acid sequences were quite homologous (more than 93%), but slightly less so between subgroups (almost 85%). However, the serotype-4 sequences differed from those of serotypes 1, 2, 3, 5, 6, 7 and 8 (less than 50%). Determining whether these differences significantly alter the epidemiology and antigenicity will require further investigation.