• Advances in Traditional Medicine
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• v.5 no.2
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• pp.117-123
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• 2005

Dyers woad leaf (Daqingye) is a traditional Chinese medicine commonly used as anti-pyretic, anti-bacterial and anti-viral agent against infectious diseases. The Chinese Pharmacopoeia (2005) records that Dyers woad leaf should be derived from the leaves of Isatis indigotica Fort., but the leaves of Polygonum tinctorium Ait., Baphicacanthus cusia (Nees) Bremek. and Clerodendron cyrtophyllum Turcz. have also been used as substitutes of Dyers woad leaf in different regions of China. The leaf morphologies of these four species show a close resemblance, and based on their morphological appearance, it is difficult to identify them. Here, molecular genetic methods were developed as a target to identify different members of Dyers woad leaf. The 5S-rRNA spacer domain was amplified by polymerase chain reaction from genomic DNAs isolated from I. indigotica, P. tinctorium, B. cusia and C. cyrtophyllum, and the nucleotide sequences showed a great diversity. In addition, random amplification of polymorphic DNA analysis was also used to distinguish the members of Dyers woad leaf. These molecular methods could be used as a tool in authentic identification of Dyers woad leaf.

• Bulletin of the Korean Chemical Society
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• v.34 no.3
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• pp.735-742
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• 2013

Peptide nucleic acids (PNAs) that bind to complementary nucleic acid sequences with extraordinarily high affinity and sequence specificity can be used as antisense oligonucleotides against microRNAs, namely antagomir PNAs. However, methods for efficient cellular delivery must be developed for effective use of PNAs as therapeutic agents. Here, we demonstrate that antagomir PNAs can be delivered to hepatic cells by complementary DNA oligonucleotide and cationic liposomes containing galactosylated ceramide and a novel cationic lipid, DMKE (O,O'-dimyristyl-N-lysyl glutamate), through glycoprotein-mediated endocytosis. An antagomir PNA was designed to target miR-122, which is required for translation of the hepatitis C virus (HCV) genome in hepatocytes, and was hybridized to a DNA oligonucleotide for complexation with cationic liposome. The PNA-DNA hybrid molecules were efficiently internalized into hepatic cells by complexing with the galactosylated cationic liposome in vitro. Galactosylation of liposome significantly enhanced both lipoplex cell binding and PNA delivery to the hepatic cells. After 4-h incubation with galactosylated lipoplexes, PNAs were efficiently delivered into hepatic cells and HCV genome translation was suppressed more than 70% through sequestration of miR-122 in cytoplasm. PNAs were readily released from the PNA-DNA hybrid in the low pH environment of the endosome. The present study indicates that transfection of PNA-DNA hybrid molecules using galactosylated cationic liposomes can be used as an efficient non-viral carrier for antagomir PNAs targeted to hepatocytes.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.15 no.1
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• pp.38-43
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• 2012

Purpose: Epstein-Barr virus (EBV) hepatitis is a usually asymptomatic and self-limiting disease in immunocompetent patients. However, the range of severity is wide, and the serological diagnosis is typically difficult until the convalescent phase. Thus, we examined the value of plasma EBV DNA real-time quantitative polymerase chain reaction (RT-qPCR) in EBV hepatitis for the timely diagnosis and the relationship between EBV viral load and clinical severity. Methods: Sixty samples were confirmed as having EBV infection by RT-qPCR with the EBV BALF5 gene sequence. We examined the clinical characteristics of EBV hepatitis by reviewing medical records. Results: The median total duration of fever was 8 days (range: 0-13 days). The mean peak value of aspartate aminotransferase (AST) was $241{\pm}214$ U/L, and the mean peak value of alanine aminotransferase (ALT) was $298{\pm}312$ U/L. There was no correlation between the serum levels of liver enzyme and plasma EBV DNA titer ($p$=0.1) or between median total duration of fever and EBV DNA titer ($p$=0.056). The median age of the EBV VCA IgM-negative group was lower compared with the EBV VCA IgM-positive group in EBV hepatitis (2 years vs. 6 years, $p$=0.0009). Conclusion: The severity of EBV hepatitis does not correlate with circulating EBV DNA load according to our data. Furthermore, we suggest that plasma EBV PCR may be valuable in young infants in whom the results of serology test for EBV infection commonly are negative.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.7
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• pp.3381-3384
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• 2016

Background: Hepatitis B virus (HBV) is a key factor for hepatocellular carcinoma (HCC). About 350 million people are affected by chronic infection which is related to the rapid development of liver diseases as well as hepatitis, cirrhosis and hepatocellular carcinoma. Expression of tumor necrosis factor alpha (TNF-${\alpha}$) in the liver demonstrates a major genetic polymorphism which is involved in resistance or susceptibility to chronic HBV infection. Materials and Methods: In this study, two populations were studied by the sequence specific primer-polymerase chain reaction (SSP-PCR) method: HBV cases (n=409), who were HBS-Ag+, and healthy controls (n=483). Results: The results shown that the frequency of TNF-${\alpha}$ -308 G/G genotype in healthy controls (47.2%) was significantly higher than in HBV infected patients (28%) (CI = 1.29-2.61, OR = 1.83, P = 0.0004). Also TNF-${\alpha}$ -308 A/A and A/G genotype frequencies in the healthy controls were 4.6% and 48.2% and in patient group were 19.5% and 52.5% (CI = 2.23-7.12, p: 0.0001, OR: 3.94) respectively. Conclusions: We found that among Iranian people TNF-${\alpha}$ -308A allele not only has the highest genotype frequency but also it has the highest frequency in the world population. In addition, TNF-${\alpha}$-308 G/G polymorphism was associated with HBV resistance, whereas TNF-${\alpha}$-308A (A/A or A/G) polymorphism appeared to associated with chronic HBV infection. These data suggested that among the Iranian population, the -308 G/G polymorphism of TNF-${\alpha}$ gene promoter region has the potential to influence the susceptibility to HBV infection and it may be responsible for viral antigen clearance.

• The Plant Pathology Journal
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• v.30 no.2
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• pp.200-207
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• 2014

A viral disease causing severe mosaic, necrotic, and yellow symptoms on Vigna angularis var. nipponensis was prevalent around Suwon area in Korea. The causal virus was characterized as Cucumber mosaic virus (CMV) on the basis of biological and nucleotide sequence properties of RNAs 1, 2 and 3 and named as CMV-wVa. CMV-wVa isolate caused mosaic symptoms on indicator plants, Nicotiana tabacum cv. Xanthi-nc, Petunia hybrida, and Cucumis sativus. Strikingly, CMV-wVa induced severe mosaic and malformation on Cucurbita pepo, and Solanum lycopersicum. Moreover, it caused necrotic or mosaic symptoms on V. angularis and V. radiate of Fabaceae. Symptoms of necrotic local or pin point were observed on inoculated leaves of V. unguiculata, Vicia fava, Pisum sativum and Phaseolus vulgaris. However, CMV-wVa isolate failed to infect in Glycine max cvs. 'Sorok', 'Sodam' and 'Somyeong'. To assess genetic variation between CMV-wVa and the other known CMV isolates, phylogenetic analysis using 16 complete nucleotide sequences of CMV RNA1, RNA2, and RNA3 including CMV-wVa was performed. CMV-wVa was more closely related to CMV isolates belonging to CMV subgroup I showing about 85.1-100% nucleotide sequences identity to those of subgroup I isolates. This is the first report of CMV as the causal virus infecting wild Vigna angularis var. nipponensis in Korea.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.751-759
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• 1999

Plaque characteristics of porcine reproductive and respiratory syndrome (PRRS) virus isolates were examined using MARC-145 line cells. The plaque morphology of PRRS virus isolates was variable in size and heterogenic in population. Upon serial passages of the PRRS virus isolates on MARC-145 tells, heterogeneity was maintained but numbers of the large plaque size virus were increased with certain isolates. A PRRS virus isolate with variable plaque sizes was subcloned into 2 populations : small plaque ($H_S$) and large plaque ($H_L$) viruses. Growth kinetics of the subclones were then determined in MARC-145 cells, and production of the structural polypeptides was analyzed by SDS-PAGE. In a comparison of the growth kinetics, the $H_S$ virus showed higher infectivity titers during the first 48 hours but slower to reach the peak titier than $H_L$ virus did. In a nucleotide sequence comparison, differences of 4 nucleotides in open reading frames 5-6 gene were found between $H_S$ and $H_L$ viruses. Both the $H_S$ and $H_L$ clones produced 5 polypeptide bands with molecular weights of 15, 19, 26, 36 and 42 kD. The 5 bands were detected at 48 hours postinoculation (PI) with antisera to $H_L$ and another large plaque virus ($W_L$) and at 72 hours PI with $H_S$ virus antiserum. The present results demonstrate differences of biologic and molecular characteristics between the two PRRS virus plaque clones.

• The Plant Pathology Journal
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• v.28 no.2
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• pp.123-136
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• 2012

Diseases caused by begomoviruses (family Geminiviridae, genus Begomovirus) constitute a serious constraint to tropical and sub-tropical agro-ecosystems worldwide. In recent years, they have also introduced in temperate regions of the world where they have great impact and are posing a serious threat to a variety of greenhouse crops. Begomoviral diseases can in extreme cases reduce yields to zero leading to catastrophic losses in agriculture. They are still evolving and pose a serious threat to sustainable agriculture across the world, particularly in tropics and sub-tropics. Till recently, there have been no records on the occurrence of begomoviral disease in South Korea, however, the etiology of other plant viral diseases are known since last century. The first begomovirus infected sample was collected from sweet potato plant in 2003 and since then there has been gradual increase in the begomoviral epidemics specially in tomato and sweet potato crops. So far, 48 begomovirus sequences originating from various plant species have been submitted in public sequence data base from different parts of the country. The rapid emergence of begomoviral epidemics might be with some of the factors like evolution of new variants of the viruses, appearance of efficient vectors, changing cropping systems, introduction of susceptible plant varieties, increase in global trade in agricultural products, intercontinental transportation networks, and changes in global climatic conditions. Another concern might be the emergence of a begomovirus complex and satellite DNA molecules. Thorough understanding of the pathosystems is needed for the designing of effective managements. Efforts should also be made towards the integration of the resistant genes for the development of transgenic plants specially tomato and sweet potato as they have been found to be widely infected in South Korea. There should be efficient surveillance for emergence or incursions of other begomoviruses and biotypes of whitefly. This review discusses the general characteristics of begomoviruses, transmission by their vector B. tabaci with an especial emphasis on the occurrence and distribution of begomoviruses in South Korea, and control measures that must be addressed in order to develop more sustainable management strategies.

• The Plant Pathology Journal
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• v.32 no.1
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• pp.33-46
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• 2016

In this study, the full-length nucleotide sequences of four Iranian PVY isolates belonging to $PVY^N$ strain were determined. The genome of Iranian PVY isolates were 9,703-9,707 nucleotides long encoding all potyviral cistrons including P1, HC-Pro, P3, 6K1, CI, 6K2, VPg, NIa-Pro, NIb and CP with coding regions of 825, 1,395, 1,095, 156, 1,902, 156, 564, 732, 1,557 and 801 nucleotides in length, respectively. The length of pipo, embedded in the P3 cistron, was 231 nucleotides. Phylogenetic analysis showed that the Iranian isolates clustered with European recombinant NTN isolates in the N lineage. Recombination analysis demonstrated that Iranian $PVY^N$ isolates had a typical European $PVY^{NTN}$ genome having three recombinant junctions while $PVY^N$ and $PVY^O$ were identified as the parents. We used dN/dS methods to detect candidate amino acid positions for positive selection in viral proteins. The mean ${\omega}$ ratio differed among different genes. Using model M0, ${\omega}$ values were 0.267 (P1), 0.085 (HC-Pro), 0.153 (P3), 0.050 (CI), 0.078 (VPg), 0.087 (NIa-pro), 0.079 (NIb) and 0.165 (CP). The analysis showed different sites within P1, P3 and CP were under positive selection pressure, however, the sites varied among PVY populations. To the best of our knowledge, our analysis provides the first demonstration of population structure of $PVY^N$ strain in mid-Eurasia Iran using complete genome sequences and highlights the importance of recombination and selection pressure in the evolution of PVY.

• The Plant Pathology Journal
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• v.29 no.3
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• pp.274-284
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• 2013

The incidence of Broad bean wilt virus 2 (BBWV2) on red pepper was investigated using the samples obtained from 24 areas of 8 provinces in Korea. Two hundred and five samples (79%) out of 260 collected samples were found to be infected with BBWV2. While the single infection rate of BBWV2 was 21.5%, the co-infection rate of BBWV2 with Cucumber mosaic virus, Pepper mottle virus, Pepper mild mottle virus and/or Potato virus Y was 78.5%. To characterize the genetic diversity of BBWV2 Korean isolates, 7 isolates were fully sequenced and analyzed. Phylogenetic analyses revealed that BBWV2 isolates could be divided largely into two groups as Group I and Group II. Based on the partial sequence analyses, 153 selected BBWV2 isolates were subgrouped into GS-I (21.6%), GS-II (3.9%) and GS-III (56.9%). BBWV2 GS-III, which was predominant in Korea, appears to be a new combination between Group I RNA-1 and Group II RNA-2. Viral disease incidence of BBWV2 on red pepper was under 2% before 2004. However, the incidence was increased abruptly to 41.3% in 2005, 58.2% in 2006 and 79% in 2007. These rapid increases might be related with the emergence of new combinations between BBWV2 groups.

• Journal of Sericultural and Entomological Science
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• v.40 no.2
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• pp.111-116
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• 1998

To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.