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Dive into the research topics where Masato Ukita is active.

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Featured researches published by Masato Ukita.


Hepatology Research | 1998

Molecular cloning and characterization of a novel DNA virus (TTV) associated with posttransfusion hepatitis of unknown etiology

Hiroaki Okamoto; Tsutomu Nishizawa; Naomi Kato; Masato Ukita; Hiroki Ikeda; Hisao Iizuka; Yuzo Miyakawa; Makoto Mayumi

Abstract The genomic DNA of a novel virus named TT virus (TTV), associated with posttransfusion hepatitis of unknown etiology, was cloned from plasma of a blood donor with an elevated transaminase level but without serological markers of known hepatitis viruses, and its sequence of 3739 bases was determined. TTV had a density of 1.26 g/cm 3 in sucrose, which did not change after the treatment with Tween 80. The viral genome was sensitive to DNase I and Mung Bean Nuclease. Hence, TTV would be an unenveloped, single-stranded DNA virus. Two possible open reading frames in different frames were identified, capable of encoding 770 and 202 amino acids, respectively. When a partial sequence of 356 bases was compared among TTV isolates from 78 sera from blood donors and hepatitis patients, it showed considerable divergence with differences of up to 30%. Oligonucleotide primers were designed on two well-conserved regions for the detection of TTV DNA in serum and biopsied liver tissues by polymerase chain reaction. TTV DNA was detected in sera from 9 of 19 (47%) patients with fulminant hepatitis and 41 of 90 (46%) patients with chronic liver disease of unknown etiology. TTV DNA was detected in liver tissues of all the five patients tested, in titers equal or 10–100 times higher than those in the corresponding sera. These results indicate that TTV would be responsible for a part of acute and chronic liver disease of unknown etiology.


Journal of Medical Virology | 1998

Fecal excretion of a nonenveloped DNA virus (TTV) associated with posttransfusion non-A-G hepatitis

Hiroaki Okamoto; Yoshihiro Akahane; Masato Ukita; Masako Fukuda; Fumio Tsuda; Yuzo Miyakawa; Makoto Mayumi

Five patients with type B or C hepatocellular carcinoma were found to be infected with a nonenveloped DNA virus (TTV) associated with posttransfusion hepatitis of non‐A–G etiology. Paired feces and serum samples from these patients were tested for TTV DNA by polymerase chain reaction with seminested primers and their sequences were compared. TTV DNA was detected in sera from all of the patients, while it was detected in feces from three patients, including two with high viral titers in serum. When feces and serum from one patient were subjected to floatation ultracentrifugation in CsCl, TTV in feces banded at a peak density of 1.35 g/cm3 and that in serum at 1.31–1.32 g/cm3. TTV isolates in three pairs of feces and serum had the identical sequence of 222 base pairs. The excretion of TTV into feces indicates that TTV would be transmitted not only parenterally but also nonparenterally by a fecal–oral route. J. Med. Virol. 56:128–132, 1998.


The Journal of Infectious Diseases | 1999

Excretion into bile of a novel unenveloped DNA virus (TT virus) associated with acute and chronic non-A-G hepatitis

Masato Ukita; Hiroaki Okamoto; Naomi Kato; Yuzo Miyakawa; Makoto Mayumi

Recently, an unenveloped, single-stranded DNA virus named TT virus (TTV) has been reported in association with hepatitis of non-A-G etiology. Five patients with TTV viremia, who received bile drainage or cholecystectomy, were tested for TTV DNA in bile by polymerase chain reaction with heminested primers. TTV DNA was detected in bile from all patients; titers were 10-100 times higher than in serum in 4 and at a comparable level in the remaining 1 patient. TTV DNA was detected in feces, also, in 1 of the 2 patients tested. The buoyant density of TTV in bile from 1 tested patient (1.33-1.35 g/cm3) was the same as that in feces (1.32-1.35 g/cm3). TTV may be secreted via bile into feces in a transmissible form and would spread by a fecal-oral route for deep and wide penetration into the general population.


Journal of Virology | 2000

Circular Double-Stranded Forms of TT Virus DNA in the Liver

Hiroaki Okamoto; Masato Ukita; Tsutomu Nishizawa; Junichi Kishimoto; Yuji Hoshi; Hitoshi Mizuo; Takeshi Tanaka; Yuzo Miyakawa; Makoto Mayumi

ABSTRACT TT virus (TTV) is an unenveloped, circular, and single-stranded DNA virus commonly infecting human beings worldwide. TTV DNAs in paired serum and liver tissues from three viremic individuals were separated by gel electrophoresis and characterized biophysically. TTV DNAs in sera migrated in sizes ranging from 2.0 to 2.5 kb. TTV DNAs in liver tissues, however, migrated at 2.0 to 2.5 kb as well as at 3.5 to 6.1 kb. Both faster- and slower-migrating forms of TTV DNAs in the liver were found to be circular and of the full genomic length of 3.8 kb. TTV DNAs migrating at 2.0 to 2.5 kb, from either serum or liver tissues, were sensitive to S1 nuclease but resistant to restriction endonucleases, and therefore, they were single-stranded. By contrast, TTV DNAs in liver tissues that migrated at 3.5 to 6.1 kb were resistant to S1 nuclease. They migrated at 3.7 to 4.0 kb after digestion with EcoRI, which suggests that they represent circular, double-stranded replicative intermediates of TTV. When TTV DNAs were subjected to strand-specific primer extension and then amplified by PCR with internal primers, those in serum were found to be minus-stranded DNAs while those in liver tissues were found to be a mixture of plus- and minus-stranded DNAs. These results suggest that TTV replicates in the liver via a circular double-stranded DNA.


Intervirology | 1999

A Novel Unenveloped DNA Virus (TT Virus) Associated with Acute and Chronic Non-A to G Hepatitis

Hiroaki Okamoto; Tsutomu Nishizawa; Masato Ukita

In 1997, a novel DNA virus was isolated from the serum of a patient with posttransfusion hepatitis of unknown etiology in Japan, and it was named TT virus (TTV) after the initials of the index patient. TTV is a nonenveloped, single-stranded and circular DNA virus, and its entire sequence of ∼3.9 kb has been determined. For being a DNA virus, TTV has a wide range of sequence divergence, allowing the classification into at least 16 genotypes separated by a sequence difference of >30% from one another. The nucleotide sequence of the noncoding region of the TTV genome is conserved, whereas that of the coding region is highly variable. TTV strains with extremely high sequence divergence are common in the same individuals, thereby indicating a mixed infection of TTV strains of different genotypes. An association is found between hepatitis of unknown etiology and the TTV genotypes which are detectable by PCR with primers deduced from the N22 region (genotype 1) in the open reading frame 1 encoding the capsid protein. It would be important to select the primers for specific detection of the TTV genotypes associated with clinical diseases, to further evaluate the capacity of TTV to induce acute and chronic liver disease as well as extrahepatic manifestations.


Journal of Virological Methods | 1999

Determination of antibodies to TT virus (TTV) and application to blood donors and patients with post-transfusion non-A to G hepatitis in Japan

Fumio Tsuda; Hiroaki Okamoto; Masato Ukita; Takeshi Tanaka; Yoshihiro Akahane; Keiko Konishi; Hiroshi Yoshizawa; Yuzo Miyakawa; Makoto Mayumi

Recently, a nonenveloped single-stranded DNA virus named TT virus (TTV) has been reported in association with non-A to G post-transfusion as well as sporadic acute and chronic liver disease. A method was developed for the detection of antibody to TTV (anti-TTV) by means of immune precipitation and detection of TTV DNA by the polymerase chain reaction. The test serum was incubated with TTV, recovered from feces of a carrier, and after incubation, the formed immune complexes were precipitated with goat antiserum to human IgG. TTV DNA was sought for by the polymerase chain reaction in both precipitate and supernatant. The detection of TTV DNA in the precipitate, but not in the supernatant, was considered to represent anti-TTV in the test serum. Of the 44 healthy blood donors in Japan, anti-TTV was detected in one of the six (17%) with TTV DNA and 11 of the 38 (29%) without TTV DNA. In the two patients with post-transfusion non-A to G hepatitis, free anti-TTV developed as they cleared TTV in serum. Anti-TTV complexed with TTV in serum, detectable by precipitating sera with goat anti-human IgG and testing for TTV DNA, elicited while the patients had elevated alanine transaminase levels. The determination of anti-TTV would be useful for detecting resolved infection in surveys for exposure to TTV in the general population, and for establishing the mechanism of liver injury associated with TTV infection.


Archives of Virology | 2000

The entire nucleotide sequences of two distinct TT virus (TTV) isolates (TJN01 and TJN02) remotely related to the original TTV isolates

Masato Ukita; Hiroaki Okamoto; Tsutomu Nishizawa; Akio Tawara; Masaharu Takahashi; Hisao Iizuka; Yuzo Miyakawa; M. Mayumi

Summary. TT virus (TTV) has a wide range of sequence divergence by which it is classified into at least 16 genotypes. A TTV isolate of genotype 12 (TJN01) and another of genotype 13 (TJN02) were sequenced in the entire genome, and compared with the reported TTV isolates. TJN01 and TJN02 had genomic lengths of 3787 and 3794 nucleotides (nt), respectively, which were shorter by 66 and 59 nt than the prototype TTV isolate of genotype 1 (TA278). TJN01 and TJN02 shared the nucleotide sequence with TA278 merely in 53.9% and 55.2%, respectively. They possessed two major open reading frames (ORFs) and the noncoding region with a GC-rich region forming stem-loop structures, which are characteristic of TTV. However, their amino acid sequences in ORF1 were similar to that of TA278 in only 35.4 and 34.0%, respectively; TJN01 was 45.4% similar to TJN02. Comparison with TTV isolates of the same genotype identified hypervariable regions in ORF1 of TJN01 and TJN02, as in the prototype TTV of genotype 1. However, quasispecies were barely observed in them. Furthermore, sequences of hypervariable regions scarcely changed during 2–5.5 years in both TJN01 and TJN02. These results indicate that TTV of genotypes 12 and 13 are much different from the prototype TTV of genotype 1.


Journal of Medical Virology | 1999

Effect of interferon on a nonenveloped DNA virus (TT virus) associated with acute and chronic hepatitis of unknown etiology

Yoshihiro Akahane; Minoru Sakamoto; Yoshiki Miyazaki; Shunichi Okada; Taisuke Inoue; Masato Ukita; Hiroaki Okamoto; Yuzo Miyakawa; Makoto Mayumi

An unenveloped DNA virus named TT virus (TTV) has been reported in association with acute and chronic hepatitis of unknown etiology. The effect of interferon on TTV was evaluated in the patients with chronic hepatitis C who were coinfected with TTV. TTV DNA was determined by a polymerase chain reaction with heminested primers in the 96 patients with chronic hepatitis C who received interferon‐α (516 million units in 26 weeks) and followed for 24 months thereafter. TTV DNA was detected in 31 (32%) patients before therapy. TTV DNA became undetectable during interferon therapy and remained absent in 14 (45% of the 31 patients) through 24 months thereafter. The four patients with pretreatment TTV DNA titer ≥103/ml did not respond. These results indicate that TTV is sensitive to interferon, and the response would be inversely correlated with pretreatment viral titers. J. Med. Virol. 58:196–200, 1999.


Transfusion | 1999

Infection by an unenveloped DNA virus associated with non‐A to ‐G hepatitis in Japanese blood donors with or without elevated ALT levels

Keiichi Itoh; Kazuya Hirakawa; Hiroaki Okamoto; Masato Ukita; Hidenori Tanaka; Naoto Sawada; Fumio Tsuda; Yuzo Miyakawa; M. Mayumi

BACKGROUND: An unenveloped, single‐stranded DNA virus named TT virus has been found in association with elevated alanine aminotransferase (ALT) levels in recipients of transfusions and has been detected frequently in patients with acute or chronic hepatitis of non‐A to ‐G etiology in Japan. DNA of the TT virus was searched for in blood donors with or without elevated ALT levels.


Virology | 1999

Marked Genomic Heterogeneity and Frequent Mixed Infection of TT Virus Demonstrated by PCR with Primers from Coding and Noncoding Regions

Hiroaki Okamoto; Masaharu Takahashi; Tsutomu Nishizawa; Masato Ukita; Masako Fukuda; Fumio Tsuda; Yuzo Miyakawa; Makoto Mayumi

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Makoto Mayumi

Gulf Coast Regional Blood Center

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Yuzo Miyakawa

Gulf Coast Regional Blood Center

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Tsutomu Nishizawa

Fukushima Medical University

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Hisao Iizuka

Gulf Coast Regional Blood Center

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Keiichi Itoh

Gulf Coast Regional Blood Center

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