Norovirus (NV) with a variety of genotypes, a member of the family Caliciviridae, causes acute nonbacterial gastroenteritis in humans. We determined the nucleotide sequence of three open reading frames (ORFs) of a NV Korean strain and characterized the genetic relationship with others. The Korean strain designated Hu/NLV/Gunpo/2006/KO was isolated from the stool specimen of a 2-year-old female suffering from gastroenteritis. By performing reverse transcription and PCR amplification, three overlapping cDNAs were synthesized and used for direct sequencing. We found that like other NVs, this strain contains three ORFs: ORF1, 5,100 bp; ORF2, 1,647 bp; ORF3, 765 bp. Of 35 NVs, ORF1 had a level of genetic diversity lower than ORF2 and ORF3, of which the C-termini of the ORF2 and ORF3 showed a relatively high degree of genetic diversity. Phylogenetic analyses indicated that the Korean strain belonged to genogroup II, with Saitama U1, Gifu'96, Mc37, and Vietnam 026 being formed a single genetic cluster. The nucleotide sequence information of three ORFs of a NV Korean isolate will be useful not only for the development of a diagnostic tool and understanding of genetic relationship, but also provide important basic information for the functional analysis of their gene products.
We have determined and analyzed the full-length cDNA sequence of a coxsackievirus B3 (CVB3) Korean isolate (CVB3-Korea/97) which has been known as a general human pathogen. The whole genome contains 7,400 nucleotides and has a single large open reading frame with 6,555 nucleotides that encodes a potential polyprotein precursor of 2,185 amino acids. The genome also contains a 5' non-coding region (NCR) of 741 bases and a 3' NCR of 104 bases followed by poly(A) tail. Sequence homologies of nucleotides and deduced amino acids between the CVB3-Korea/97 strain and the prototype (Nancy strain) were 81.7% and 91.5%, respectively. The genes encoding the functional proteins including viral protease and RNA dependent RNA polymerase showed higher homology than those encoding the structural proteins. We have further analyzed the sequences of 5' NCR, VP1 and VP2 of CVB3-Korea/97, which are known as cardiovirulent determining factors at the nucleotide and amino acid levels. Although the CVB 3-Korea/97 strain was isolated from an aseptic meningitis patient without cardiomyopathy, its 234th nucleotide and 165th amino acid were uracil and Asn as same as those of other cardiovirulent strains one. However, the 155th amino acid of VP1, which closely associated with cardiovirulence, was replaced with $Arg^{155}$ by single nucleotide substitution from $A^{2916}$ to $T^{2916}$. Moreover, additional amino acid substitutions were observed in the flanking region of $Asp^{155}$. Taken together, amino acid(s) substitution in VP1 may playa critical role in determining cardiovirulence of the CVB3-Korea/97 strain rather than individual nucleotide replacements in the 5' NCR and/or an amino acid substitution in VP2.
A few SV4O-transformed human cells such as SV8O are potentially tumorigenic but rejected by athymic hosts. However, one cell line in this group (W118IVA-2) is known to be fully tumorigenic. Two clones were obtained after the injection of W118IVA-2, of which NW1SC1-1 was tumorigenic but NW18C1-2 was not in nude mice. As examined by Southern blot analysis, NW18C1-1 appears to contain more copy number of SV40 sequences than NW18C1-2 does. However, it was unable to demonstrate that this difference elicits the tumorigenicity in NW18C1-1 but not in NW18C1-2. Therefore, the latter clone was tested if it expresses SV40 early genes to produce large T as well as small t antigens using indirect immunofluorescent assay and immunoprecipitation. In addition, mouse NIH3T3 cells were transfected with the cellular DNA of NW1SC1-2 as well as that of NW18C1-1 to examine if the viral genomes in the clones can make the nontransformed cells to acquire malignant growth potential in vivo. The transformed cells expressed large T antigen and became tumorigenic. Thus, the transforming functions of NW1SC1-2 cell appers to be intact. These results clearly suggest that the inability of NW18C1-2 cell to form tumor in nude mice is not because they are inherently nontumorigenic. However, the possibility that the interaction of SV40 with its host differs in these clones can not he ruled out.
Cristina, San Juan Lopez;Marta, Casado Martin;Mercedes, Gonzalez Sanchez;Almudena, Porcel Martin;Alvaro, Hernandez Martinez;Luis, Vega Saenz Jose;Tesifon, Parron Carreno
Clinical and Molecular Hepatology
/
v.24
no.4
/
pp.384-391
/
2018
Backgrounds/Aims: The objective of our study was to determine the epidemiological, laboratory, and serological characteristics of patients with chronic hepatitis B virus (HBV) infection and normal transaminases. The study also aimed to evaluate liver damage by measuring the liver fibrosis (LF) grade and to identify possible factors associated with the presence of fibrosis. Methods: A retrospective observational study was conducted in patients with chronic HBV infection and classified as inactive carriers or immune-tolerant. Epidemiological variables of age, sex, immigrant, alcohol consumption, and body mass index (BMI), as well as virological variables (HBV DNA) and transaminase level were collected throughout the follow-up. The LF grade was evaluated by transient elastography. The cutoff value for significant fibrosis (SF) was liver stiffness ${\geq}7.9kPa$. Results: A total of 214 patients were included in the analysis, and 62% of them had a BMI ${\geq}25kg/m^2$. During follow-up, 4% of patients showed transaminase elevation (<1.5 times normal). Most patients had a viral DNA level <2,000 IU/mL (83%). Data on LF were available in 160 patients; of these, 14% had SF, 9% F3, and 6% F4. The variables associated with the presence of SF were transaminase alteration during follow-up, as 23% of patients with SF had elevated transaminases versus 3% of patients without SF (P<0.005), and BMI, as the vast majority of patients with SF (88%) had a BMI ${\geq}25kg/m^2$ versus 56% of patients without SF (P<0.05). Conclusions: In patients with chronic HBV infection and normal transaminases, liver damage does not seem to be related to DNA levels, alcohol consumption, or immigrant status. SF seems to be associated with transaminase alteration during follow-up and elevated BMI. It is therefore recommended to measure LF grade with validated non-invasive methods in such patients.
Chicken Mx protein (cMx) induced interferon (IFN) is an antiviral protein to inhibit replication of RNA virus, particularly negative stranded RNA virus, through blockage of transfortation of viral RNA and proteins. In order to determine antiviral effects of cMx from different MHC haplotype chicken, we characterized cMx gene by studying on nucleotide sequencing, antiviral effects to Newcastle disease virus, VSV and MDV, and transcription activities. Three types of eMx genes (2,118 bp) were detected from the different MHC haplotype chickens [B19 (N), B15(F) and B21 (GSP)] chickens, which have showed different susceptible to Marek's disease (MD). Several amino acid substitutions were showed in the cMx. The amino acid 548 and 631 in the cMxs from N and F, chickens susceptible to MD, was Val and Asn which was important on antiviral effects, and showed in resistant cMx. Those in the cMx from GSP, chicken resistant to MD, were same that showed in susceptible cMx. Though every cMx transactivated the expression of the reporter gene, the transcription activation by resistant cMx from N and F was lower compared to that by susceptible cMx from GSP. The decease of the cell growth in the resistant cMx cloned cells was seen in comparison with another cMx clone cells. Replication of NDV and VSV was suppressed in the clones with resistant cMx from N and F. NMx258-transducted cells lack of antiviral effects, and NMx437 or NMx646-transducted cells was showed 60% of antiviral effects compared to NMx705. Mean death time (MDT) and hemaggutination (HA) titer to NDV was long and low in the eggs of N and F lines, but short and high in the egg of GSP line. Interestingly, strong suppression to NDV was observed in the clone with N-Mx and in the eggs of N line. However, the effects of Mx for replication of vvMDV1 have not been. Thus, resistant types of cMx, N- and F-Mx, have showed the anti-viral effects to only RNA virus including NDV and VSV, but not to DNA virus. Antiviral effects of cMx were required whole length of amino acid including Val and Asn in amino acid 548 and 631.
Alfalfa mosaic alfamoviruses(AIMV) were isolated from infected potato (Solanum tuberosum) and azuki bean (Paseolus angularis) in Korea. Two AIMV isolated from potatoes were named as strain KR (AIMV-KR1 and KR2) and AIMV isolated from azuki bean was named as strain Az (AIMV-Az). Each isolated AIMV strain was characterized by using their host ranges, symptom developments, serological relations and nucleotide sequence analysis of coat protein (CP) gene. Strains KR1, KR2, and Az were readily transmitted to 20 of 22 inoculated plant species including bean, cowpea, tomato, tobacco, and potato. AIMV-KR1 and KR2 produced the typical symptoms like chlorotic or necrotic spots in Chenopodium quinoa and Solanum tuberosum cv. Superior. AIMV-Az caused bright yellow mosaic symptom and leaf malformation in Nicotiana glauca, which were different from the common mosaic symptom caused by AIMV-KR1 and KR2. Electron microscope observation of purified virus showed bacilliform virions containing a single-stranded plus-strand RNAs of 3.6, 2.6, 2.0 and 0.9 kbp in length, respectively, similar in size and appearance to those of Alfamovirus. In SDS-PAGE, the coat protein of the two viruses formed a consistent band that estimated to be about 24kDa. The CP genes of the AIMV strains, KR1, KR2, and Az have been amplified by RT-PCR using the specific primers designed to amplify CP gene from viral RNA-3, cloned and sequenced. Computer aided analysis of the amplified cDNA fragment sequence revealed the presence of a single open reading frame capable of encoding 221 amino acids. The nucleotide and peptide sequence of viral CP gene showed that strain KR1, KR2, and Az shared highest nucleotide sequence identities with AIMV strain 425-M at 97.7%, 98.2%, and 97.2%, respectively. CP gene sequences of two strains were almost identical compared with each other. Altogether, physical, serological, biological and molecular properties of the purified virus.
So, Kyeong A;Kim, Seon Ah;Lee, Yoo Kyung;Lee, In Ho;Lee, Ki Heon;Rhee, Jee Eun;Kee, Mee Kyung;Cho, Chi Heum;Hong, Sung Ran;Hwang, Chang Sun;Jeong, Mi Seon;Kim, Ki Tae;Ki, Moran;Hur, Soo Young;Park, Jong Sup;Kim, Tae Jin
Obstetrics & gynecology science
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v.61
no.6
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pp.662-668
/
2018
Objective This study was to identify the risk factors for cytological progression in women with atypical squamous cells of undetermined significance (ASC-US) or low-grade squamous intraepithelial lesions (LSIL). Methods We analyzed data from women infected with the human papillomavirus (HPV) who participated in the Korean HPV cohort study. The cohort recruited women aged 20-60 years with abnormal cervical cytology (ASC-US or LSIL) from April 2010. All women were followed-up at every 6-month intervals with cervical cytology and HPV DNA testing. Results Of the 1,158 women included, 654 (56.5%) and 504 (43.5%) women showed ASC-US and LSIL, respectively. At the time of enrollment, 143 women tested positive for HPV 16 (85 single and 58 multiple infections). Cervical cytology performed in the HPV 16-positive women showed progression in 27%, no change in 23%, and regression in 50% of the women at the six-month follow-up. The progression rate associated with HPV 16 infection was higher than that with infection caused by other HPV types (relative risk [RR], 1.75; 95% confidence interval [CI], 1.08-2.84; P=0.028). The cytological progression rate in women with persistent HPV 16 infection was higher than that in women with incidental or cleared infections (P<0.001). Logistic regression analysis showed a significant relationship between cigarette smoking and cytological progression (RR, 4.15; 95% CI, 1.01-17.00). Conclusion The cytological progression rate in HPV 16-positive women with ASC-US or LSIL is higher than that in women infected with other HPV types. Additionally, cigarette smoking may play a role in cytological progression.
In this review, recent foodborne outbreaks caused by viruses in the Republic of Korea (2010-2019) were analyzed. The human norovirus was found to be the major foodborne virus causing an average of 94.9% of the viral outbreaks. Reverse-transcription polymerase chain reaction (PCR) with electrophoresis has been widely used to detect viruses, but several rapid detection methods, including real-time PCR, multiplex PCR, and quantum dot assay, have also been suggested. For norovirus inactivation studies, surrogates such as murine norovirus and feline calicivirus have been widely used to identify the reduction rate owing to the limitations in laboratory cultivation. Conversely, direct cell infection studies have been conducted for other foodborne viruses such as adenovirus, astrovirus, rotavirus, and hepatitis A or E virus. Moreover, virucidal mechanisms using various physical and chemical treatments have been revealed. These recent studies suggest that rapid in situ detection and effective control are valuable for ensuring food safety against viral infections.
Purpose: Interferon is a widely used treatment for chronic hepatitis B in children. However, additional treatment options are needed because more than 50% of hepatitis B patients are unresponsive to interferon. Although lamivudine is widely used to treat hepatitis B, there are few studies on the effect of lamivudine in hepatitis B patients unresponsive to interferon. Methods: Eight interferon unresponsive patients (6 males and 2 females) were treated with lamivudine (3 mg/kg/day, maximum 100 mg/day) from 6~12 months after interferon treatment was discontinued among 33 children with chronic hepatitis B. They were treated with interferon (interferon ${\alpha}$-2b, 10 MU/$m^2$ or pegylated interferon $1.5{\mu}g/kg$) for 6 months from January 2000 to December 2007 at the Pusan National University Hospital. The medical records were analyzed retrospectively. Results: The age at treatment with interferon and lamivudine was 4.9${\pm}$3.1 and 6.1${\pm}$3.2 years, respectively. The serum ALT level before treatment with interferon was 148.1${\pm}$105.8 IU/L and the log HBV-DNA PCR mean value was 6.95${\pm}$0.70 copies/mL. The serum ALT level after treatment with interferon was 143.1${\pm}$90.4 IU/L and the log HBV-DNA mean PCR value was 6.46${\pm}$2.08. HBeAg negativization occurred in 2 patients. For all patients, normalization of the serum ALT levels and HBeAg seroconversion (except 2 patients with HBeAg negativization) occurred at 7.4${\pm}$2.1 and 7.9${\pm}$2.1 months respectively after lamivudine treatment. The HBV-DNA PCR became negative in 7 patients (87.5%) at 2.4${\pm}$2.8 months. Complete response was achieved in 7 patients and no recurrence was observed in 2 patients for 3 years after the completion of treatment. Five patients are still under treatment for a mean treatment duration of 24.4${\pm}$9.1 months. In one patient, viral breakthrough occurred and the treatment was stopped. Conclusion: The number of patients was small, however, lamivudine treatment in patients with chronic hepatitis B who were unresponsive to interferon was highly effective.
Kim, Hyun;Chae, Hee-Bok;Jeon, Won-Joong;Park, Seon-Mee;Youn, Sei-Jin;Eun, Jong-Ryul;Lee, Heon-Ju
Journal of Yeungnam Medical Science
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v.25
no.1
/
pp.31-40
/
2008
Background/Aims : Entecavir is a synthetic nucleoside analogue, cyclopentyl guanine nucleoside, which has a potent antiviral effect and the least viral breakthrough in hepatitis B virus (HBV) replication. Entecavir has been available in Korea since 2007 but there are few reports on its effects. The aim of this study was to evaluate the virological response (VR) and biochemical response (BR) to entecavir in HBV patients at 3, 6 and 9 months after treatment with entecavir. Materials and Methods : Thirty-three chronic hepatitis B patients who took entecavir for at least 9 months were enrolled. We investigated VR and BR by retrospectively reviewing medical records. Patients who satisfied the following criteria were chosen: 1) initial alanine aminotransferase (ALT) levels = 1.5upper limit of normal (ULN) and 2) initial HBV DNA levels = $5\;log_{10}\;copies/ml$. We measured ALT levels every 3 months until month 9. HBV DNA was measured every 2 or 3 months by polymerase chain reaction (PCR) method. Results : Most patients taking entecavir showed good BR (ALT < 40 IU/L). The BR rates were 61%, 73% and 67% at months 3, 6 and 9, respectively. VR (HBV DNA < $5\;log_{10}\;copies/ml$ or 2 log lower than initial HBV DNA) rates were 82%, 91% and 91% at months 3, 6 and 9, respectively. Undetectable HBV DNA (HBV DNA < 4 log10 copies/ml) rates were 49%, 73% and 85% at months 3, 6 and 9, respectively. Two patients presented with virological breakthrough without adverse effects until month 9. Conclusions : Entecavir showed good BR and VR from month 3 and these effects continued through the 9-month observation period. This suggests that entecavir is also a good choice for the first line treatment of chronic hepatitis B (CHB). Further studies are needed to determine the long-term efficacy and drug resistance of entecavir in Korean CHB patients.
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