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O Nakagomi

Publications and source records attributed to O Nakagomi.

At least 19 recordsLinked to original sources

A lack of significant association between the electropherotype or G-serotype of the infecting strain and disease severity of rotavirus gastroenteritis.

Despite many previous studies, the question has not been settled as to whether some human rotavirus strains are more virulent than others. Since disease severity is most clearly reflected by the hospitalization status of the infected children, we examined whether there was any difference in the distribution of dominant strains between inpatient and outpatient groups. The study population comprised 763 children with acute diarrhea who were treated at a general hospital in Honjo City, Akita, Japan, during 1986-1997. Rotaviruses from stool specimens were classified into 77 electropherotypes using polyacrylamide gel electrophoresis. A single dominant strain or two co-dominant strains circulated simultaneously with some infrequent strains in most rotavirus seasons. Over the 11 rotavirus seasons, there was no significant difference in the relative frequencies of 15 rotavirus strains between the inpatient and the outpatient groups when strains of rotavirus were defined by their electropherotypes in polyacrylamide gel electrophoresis. However, infection with one G1 strain that co-dominated with a G4 strain carrying an identical electropherotype except the VP7 gene resulted in a statistically significantly reduced risk of hospitalization. There was no significant difference in the relative frequencies of four major G-serotypes or long/short RNA pattern. We conclude that the virulence or disease-causing potential of human rotavirus is not substantially different in the majority of strains.

Adolescent↗

A high incidence of intussusception in Japan as studied in a sentinel hospital over a 25-year period (1978-2002).

The development of second-generation rotavirus vaccines requires knowledge of baseline incidence rates for intussusception in infants prior to vaccine introduction. To obtain such estimates we reviewed clinical records in a hospital that served as the major provider of paediatric beds in a local community in the northern part of Japan. During the 25-year period (1978-2002), there were 91 hospitalizations due to radiologically confirmed intussusception in children <5 years of age, of which 45% were <1 year of age. Assuming that all children with intussusception in the area had been admitted to this hospital, there were an average of 185 and 78 hospitalizations per 100000 person-years for children <1 year old and 5 years old respectively. There was period-to-period variability with no long-term secular trend in the incidence of intussusception. The incidence rate in Japan was among the highest thus far reported, providing further evidence of geographic variability.

Child, Preschool↗

Isolation, serologic and molecular characterization of the first G3 caprine rotavirus.

We isolated a rotavirus in cell culture, named the GRV strain, from a stool specimen of a Korean goat with diarrhea, and performed an in-depth characterization. At various passage levels in cell culture, the GRV strain retained its pathogenicity for goat kids, thereby for the first time establishing that a caprine rotavirus can cause diarrhea in goat kids. The GRV strain grew to a high titer and agglutinated group O human erythrocytes. The GRV VP7 protein was 96% identical with the RRV (simian rotavirus) and R2 (lapine rotavirus) VP7 proteins, and slightly less similar to the SA11 (simian rotavirus) and HCR3 (feline/canine-like human rotavirus) VP7 proteins. The GRV VP4 protein was 93% identical with the RRV VP4 (P[3]) and 90% identical with the SA11 VP4 (P[2]). However, phylogenetic analysis including more VP4 sequences from representative P[3] strains unambiguously placed the GRV VP4 in the cluster of P[3] VP4s. A high level of two-way cross neutralization with RRV substantiated that GRV was a G3P5[3] strain, thus identifying GRV as the first caprine rotavirus with such a phenotype. The GRV NSP4 sequence belonged to the AU-1 allele, as does the RRV NSP4 sequence. Genetic analysis by RNA-RNA hybridization revealed that the overall genomic RNA constellation of the GRV strain was unique among mammalian rotavirus genogroups and that it was almost equally related to, yet distant from, simian rotavirus RRV, feline/canine rotavirus FRV64 (or CU-1), feline/human rotavirus FRV-1 (or AU-1), and lapine rotavirus R2. The availability of the GRV strain will further expand our limited knowledge of caprine rotaviruses.

Alleles↗

Genomic RNA constellation of recently emerging serotype G14 equine rotavirus strains in Japan that is highly homologous with prototype G3 and G14 strains previously identified in the United States of America.

Serotype G14 was once considered to be uncommon among equine rotaviruses. While it sporadically emerged in some parts of the world, serotype G14 became the dominant G serotype among rotaviruses detected in foals with diarrhea in Japan in the late 1990s. However, it is not known how such recently emerging G14 rotaviruses are related in their overall genomic RNA constellation to prototype G14 strain identified earlier in the United States of America or how they were generated and why they have dominated over G3 equine rotaviruses. Genogrouping by RNA-RNA hybridization revealed that recently emerging serotype G14 equine rotavirus strains had an overall genomic RNA constellation that was highly conserved not only with contemporary and earlier G3 strains in Japan but also with prototype G3 and G14 strains previously identified in the United States of America. Japanese G14 rotavirus strains are likely to have originated form a VP7 gene substitution reassortant that had been formed earlier in the United States of America on the background of the then dominant G3 equine rotavirus.

Animals↗

Characterization of serotype G9 rotavirus strains isolated in the United States and India from 1993 to 2001.

The emergence of rotavirus serotype G9 as a possible fifth globally common serotype in the last decade, together with its increasing detection in association with various genome constellations, raises questions about the origins and epidemiological importance of recent G9 isolates. We examined a collection of 40 G9 strains isolated in the United States from 1996 to 2001 and in India since 1993 to determine their VP7 gene sequences, P types, E types, subgroup specificities, and RNA-RNA hybridization profiles. With the exception of two U.S. strains, all of the study strains shared high VP7 gene sequence homology (<2.5% sequence divergence on both the nucleotide and amino acid levels) and were more closely related to other recent isolates than to the first G9 strains isolated in the 1980s. The VP7 gene sequence and RNA-RNA hybridization profiles of the long-E-type strains showed greater variation than the short-E-type strains, suggesting that the latter strains are the result of a relatively recent reassortment event of the G9 VP7 gene into a short-E-type lineage. No evidence for reassortment of genes other than VP4 and VP7 between major human rotavirus genogroups was observed. Except for Om46 and Om67, which formed a distinct clade, phylogenetic analysis showed that most of the study strains grouped together, with some subgroups forming according to genetic constellation, geographic location, and date of isolation. The high potential of G9 strains to generate different P and G serotype combinations through reassortment suggests that it will be important to determine if current vaccines provide heterotypic protection against these strains and underscores the need for continued surveillance for G9 and other unusual or emerging rotavirus strains.

Amino Acid Sequence↗

Characterization of nontypeable rotavirus strains from the United States: identification of a new rotavirus reassortant (P2A[6],G12) and rare P3[9] strains related to bovine rotaviruses.

Among 1316 rotavirus specimens collected during strain surveillance in the United States from 1996 to 1999, most strains (95%) belonged to the common types (G1 to G4 and G9), while 5% were mixed infections of common serotypes, rare strains, or not completely typeable. In this report, 2 rare (P[9],G3) and 2 partially typeable (P[6],G?; P[9],G?) strains from that study were further characterized. The P[6] strain was virtually indistinguishable by hybridization analysis in 10 of its 11 gene segments with recently isolated P2A[6],G9 strains (e.g., U.S.1205) from the United States, but had a distinct VP7 gene homologous (94.7% a.a. and 90.2% nt) to the cognate gene from P1B[4],G12 reference strain L26. Thus, this serotype P2A[6],G12 strain represents a previously unrecognized reassortant. Three P3[9] strains were homologous (97.8-98.2% aa) in the VP8 region of VP4 to the P3[9],G3 feline-like reference strain AU-1, but had a high level of genome homology to Italian bovine-like, P3[9],G3 and P3[9],G6 rotavirus strains. Two of the U.S. P3[9] strains were confirmed to be type G3 (97.2-98.2% VP7 aa homology with reference G3 strain AU-1), while the other was most similar to Italian bovine-like strain PA151 (P3[9],G6), sharing 99.0% a.a. homology in VP7. Cross-neutralization studies confirmed all serotype assignments and represented the first detection of these rotavirus serotypes in the United States. The NSP4 genes of all U.S. P3[9] strains and rotavirus PA151 were most closely related to the bovine and equine branch within the DS-1 lineage, consistent with an animal origin. These results demonstrate that rare strains with P and G serotypes distinct from those of experimental rotavirus vaccines circulate in the United States, making it important to understand whether current vaccine candidates protect against these strains.

Animals↗

Molecular characterization of serotype G2 and G3 human rotavirus strains that have an apparently identical electropherotype of the short RNA pattern.

The literature is conflicting whether or not rotavirus strains with different G serotype have an identical electropherotype. This is a contentious but an important issue because large parts of molecular epidemiological studies of rotaviruses have been based on the conception that a single strain of rotavirus can be defined by a single electropherotype. Here, we examined in detail by reverse-transcription PCR genotyping, electropherotyping, sequencing, and genogrouping using RNA--RNA hybridization three human rotavirus strains isolated in India that had apparently identical electropherotypes although one strain was typed as P[4], G3 while the other two typed as P[4], G2. These three strains showed an identical electropherotype on 7.5% and 12.5% polyacrylamide gels, but co-electrophoresis on a 10% gel demonstrated that segment 8 of the P[4], G3 strain migrated more slowly than the cognate segment of the P[4], G2 strains. Genogrouping assay and nucleotide sequencing provided evidence for the hypothesis that the P[4], G3 stain was an intergenogroup reassortant in which a P[4], G2 strain of the DS-1 genogroup had acquired the VP7 gene from an yet-unidentified concurrently circulating G3 strain. While electropherotyping remains a valuable asset for molecular epidemiology of rotaviruses, this study underscores the importance of co-electrophoresis under different electrophoretic conditions when pinpointing subtle differences.

Antigens, Viral↗

Winter seasonality and rotavirus diarrhoea in adults.

We investigated the aetiological role of group A rotavirus in adults with acute diarrhoea in a 4-year prospective study. Of 683 patients with acute diarrhoea, 97 (14%) shed rotavirus as a sole agent, whereas six (5%) of 115 patients without diarrhoea shed rotavirus. Half of patients with rotavirus diarrhoea required admission to hospital. Unlike rotavirus diarrhoea in children, the occurrence of rotavirus-positive cases did not show a significant winter seasonality. Rotavirus infection should be included in the differential diagnosis of diarrhoeal diseases in adults.

Adult↗

Group C rotavirus NSP4 induces diarrhea in neonatal mice.

Nonstructural glycoprotein NSP4 of group A rotavirus induces diarrhea in neonatal mice by functioning as an enterotoxin. Previously, our laboratory reported that the structural features of group A and group C rotavirus NSP4 proteins are well conserved despite a lack of sequence homology between group A and group C rotavirus NSP4 proteins [Horie Y, et al., Arch Virol (1997) 142: 1865-1872]. To test whether group C rotavirus NSP4 has an enterotoxigenic activity, we expressed in Escherichia coli the carboxy two-thirds (corresponding to amino acid residues 55-150) of the NSP4 protein derived from group C rotavirus strain Ehime 9301. This truncated NSP4 protein was able to induce diarrhea in 5-day-old CD-1 mice when administered intraperitoneally. Thus, group C rotavirus NSP4 acts as an enterotoxin like group A rotavirus NSP4.

Animal Population Groups↗

Direct evidence for genome segment reassortment between concurrently-circulating human rotavirus strains.

Extensive heterogeneity in electropherotypes observed among group A human rotaviruses has been considered as a result of two major mechanisms; i.e., the accumulation of point mutations and genetic reassortment between concurrently-circulating strains. However, no evidence was reported thus far indicating that any one of field isolates of rotavirus was formed by direct reassortment of concurrently circulating two parental strains. Comparison of the genome of human rotavirus specimens collected over a six year period by electropherotyping and by the sequencing of selected gene segments identified two reassortants that were generated in nature between strains circulating co-dominantly in the same epidemic season. This is the first report directly showing that at least some part of electrophoretic diversity observed among rotavirus strains was explained by genetic reassortment between strains concurrently circulating in the human population. This supports the hypothesis that genetic reassortment among co-circulating strains operates as a key mechanism for the genetic variability of rotaviruses in nature.

Base Sequence↗

Analysis of anti-rotavirus activity of extract from Stevia rebaudiana.

Anti-human rotavirus (HRV) activity of hot water extracts from Stevia rebaudiana (SE) was examined. SE inhibited the replication of all four serotypes of HRV in vitro. This inhibitory effect of SE was not reduced on the prior exposure of SE to HCl for 30 min at pH 2. Binding assay with radiolabeled purified viruses indicated that the inhibitory mechanism of SE is the blockade of virus binding. The SE inhibited the binding of anti-VP7 monoclonal antibody to HRV-infected MA104 cells. The inhibitory components of SE were found to be heterogeneous anionic polysaccharides with different ion charges. The component analyses suggested that the purified fraction named as Stevian with the highest inhibitory activity consists of the anionic polysaccharide with molecular weight of 9800, and contains Ser and Ala as amino acids. Analyses of sugar residues suggest uronic acid(s) as sugar components. It did not contain amino and neutral sugars and sulfate residues. These findings suggest that SE may bind to 37 kD VP7 and interfere with the binding of VP7 to the cellular receptors by steric hindrance, which results in the blockade of the virus attachment to cells.

Animals↗

A lack of consistent amino acid substitutions in NSP4 between rotaviruses derived from diarrheal and asymptomatically-infected kittens.

Nonstructural glycoprotein NSP4 of group A rotavirus has recently been shown to be a viral enterotoxin, inducing diarrhea in neonatal mice. Literature is conflicting as to whether there is any consistent amino acid substitution between virulent (or symptomatic) and attenuated (or asymptomatic) rotavirus strains. We have sequenced and compared the NSP4 sequences derived from a total of 10 geographically- and serologically-related feline rotavirus strains from both diarrheal and asymptomatically-infected kittens. These NSP4 sequences were closely related to each other and there were differences at 19 amino acid residues, but none was segregated according to whether the strain was isolated from a diarrheal kitten or not. Thus, this study failed to lend support to the contention that mutations in NSP4 play a significant role in the pathogenesis of rotavirus diarrhea. Involvement of other genes may explain the outcome of infection in cats from which these 10 feline rotaviruses were isolated.

Amino Acid Sequence↗

Much higher risk of premalignant and malignant cervical diseases in younger women positive for HPV16 than in older women positive for HPV16.

Human papillomavirus (HPV), particularly HPV16, is strongly associated with premalignant lesions of the uterine cervix and cervical cancer. However, HPV infection is a common sexually transmitted disease and only a few women develop cervical cancer. Although the presence of HPV and abnormal cytology are independent risk factors for cervical diseases, implementing both tests on every woman is argued not to be recommended mainly in terms of cost-effectiveness. During a 20-month period between October, 1994, and May, 1996, cervical swabs from 207 women who were referred for colposcopy because of cervical dyskaryosis (Papanicolaou class IIIa or higher) were examined by PCR for the presence of HPV16. When these women were divided into two groups; i.e., group A consisting of women who were 44 years old or younger (n = 111), and group B consisting of women who were 45 years old or older (n = 96), the risk of having premalignant and malignant cervical diseases upon infection with HPV16 was approximately 8 times higher in group A than in group B. Thus, we conclude that HPV-testing should be implemented on every young woman with an abnormal Papanicolaou smear test.

Adult↗

Molecular and serologic characterization of novel serotype G8 human rotavirus strains detected in Blantyre, Malawi.

During a 2-year study of diarrhea among children in Blantyre, Malawi, greater than 50% of rotavirus strains genotyped by using reverse transcription-polymerase chain reaction possessed previously unrecognized combinations of the neutralization proteins VP7 and VP4. Serotype G8 rotaviruses, which have been identified recently in several African countries, were found to possess P[4] or P[6] VP4 genotype specificity. Two of these short electropherotype rotaviruses were further investigated: these comprised a P[6], G8 representative strain (MW23) and a P[4], G8 representative strain (MW333). The VP7 gene sequences of both strains exhibited greatest homology to human and animal serotype G8 rotaviruses. Sequence analysis of the VP4 gene of MW23 indicated closest identity to the P2A[6], G9 strain US1205 from the United States. The VP4 gene of MW333 was most closely related to the P[4], G12 strain L26 isolated in the Philippines and the Australian P[4], G2 strain RV-5. The NSP4 gene sequences of both strains were classified in NSP4 genetic group I. RNA-RNA hybridization demonstrated that each of these two strains is related to the DS-1 genogroup of human rotaviruses. Subgroup analysis and virus neutralization confirmed complete antigenic characterization of MW23 as subgroup I, P2A[6], G8 and MW333 as subgroup I, P1B[4], G8. The similarity of the VP7 gene sequences of the prototype strains described in this report to bovine serotype G8 rotaviruses suggests that they may represent human/bovine reassortant viruses.

Animals↗

Human rotavirus HCR3 possesses a genomic RNA constellation indistinguishable from that of feline and canine rotaviruses.

Infection and spread of group A rotaviruses under natural conditions are mostly limited to one animal host species. However, rare molecular evidence exists for interspecies transmission by whole virions of animal rotaviruses to humans. Human rotavirus strain HCR3, which was isolated in 1984 from a healthy infant in Philadelphia, U.S.A. was shown by RNA-RNA hybridization to form 11 hybrid bands with feline rotavirus strain FRV64 and canine rotavirus strains CU-1 and K9, but not with rotaviruses commonly found in humans. Thus, HCR3 was concluded to be originally a rotavirus circulating in cats and dogs and accidental interspecies transmission by whole virions to humans was likely to have occurred in the past.

Animals↗

The relative frequencies of G serotypes of rotaviruses recovered from hospitalized children with diarrhea: A 10-year survey (1987-1996) in Japan with a review of globally collected data.

Since rotavirus vaccines aim to protect children from severe diarrhea, knowledge of the prevailing G serotypes among rotaviruses from hospitalized children is essential. Thus, we determined the G serotypes of rotaviruses collected from children with acute diarrhea in a local referral hospital in Akita, Japan, over the 10-year period between January 1987 and December 1996. Based on the assumption that rotaviruses with an identical electropherotype possess the same G serotype, the G serotypes of 488 rotavirus-positive specimens that were classified into 63 electropherotypes were determined by enzyme-linked immunosorbent assay with a supplementary use of G typing by reverse transcription-PCR. The relative frequencies over the 10-year period were 77.0 (G1), 14.5 (G2), 2.7 (G3) and 5.3% (G4), leaving the possibility that only 0.4% had G serotypes uncommon to human rotaviruses. Of 24,050 rotaviruses extracted by reviewing 63 serotyping studies in literature, the relative frequencies of the four major G serotypes were 50.6 (G1), 9.3 (G2), 7.2 (G3) and 11.6% (G4). As to uncommon G serotypes, only 0.9% were described as serotypes other than G1-4, and our estimate for potential uncommon serotypes were at most 8.1%. Thus, both this long-term study focusing on the rotaviruses only from severe cases in a single hospital in Japan and the global review of G serotypes published to date indicate that the primary target of any rotavirus vaccines should be rotaviruses possessing serotypes G1-4.

Child↗