• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.6 no.2
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• pp.140-151
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• 2003

Purpose: We tried to evaluate the long term efficacy and positive predictive factors of interferon-alpha treatment in children with chronic hepatitis B. Methods: The study population included 113 children who received interferon therapy between May 1982 and July 2002 (20 years) for chronic hepatitis B in Department of Pediatrics, Yonsei University College of Medicine. Male to female ratio was 2.3 : 1 and the mean age at diagnosis was $11.1{\pm}4.1$ years old. Response to treatment was defined as normalization of alanine aminotransferase (ALT), disappearance of HBeAg and HBV-DNA Eighty two children responded while 32 did not. Interferon-alpha was given intramuscularly for 6 months at a dosage of $3{\times}10^6$ unit, 3 times weekly. In relapsed cases, lamivudine or interferon retreatment was done. Results: Seroconversion rate was 77.0% in terms of HBeAg, 74.3% in terms of HBV-DNA, and 80.5% in terms of ALT normalization after treatment. Seroconversion rate of both HBeAg and HBV-DNA was 72.6%. Analyzed by life table method, the effect of the treatment had been maintained over 10 years after cessation of therapy. Pre-treatment ALT level was the only significant positive predictive factor of response. Eleven cases (13.4%) relapsed, and 2 out of 3 showed response when treated with lamivudine and 1 out of 3 with interferon retreatment. Conclusion: Interferon-alpha showed significant efficacy in the treatment of chronic hepatitis B in our study. Further studies about the effect of interferon therapy on complications of hepatitis such as hepatocarcinoma, cirrhosis are warranted.

• Tuberculosis and Respiratory Diseases
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• v.51 no.4
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• pp.315-324
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• 2001

Background : IL-8 is a potent chemotactic cytokine that plays an important role in the host defense mechanism against M. tuberculosis by recruiting inflammatory cells to the site of the infection. Lung epithelial cells, as well as alveolar macrophages are known to produce IL-8 in response to M. tuberculosis. IL-8 gene expression is mainly regulated on the level of transcription by NF-${\kappa}B$. This study investigated whether or not A549 cells produce IL-8 in NF-${\kappa}B$ dependent mechanism in response to macrophages phagocytosing M. tuberculosis. Methods : Peripheral blood monocytes that were obtained from healthy donors were cultured for 24 h with M. tuberculosis and a conditioned medium(CoMTB) was obtained. As a negative control, the conditioned medium without M. tuberculosis (CoMCont) was used. A549 cells were stimulated with M. tuberculosis, CoMCont and CoMTB and the IL-8 concentration in the culture media was measured by ELISA. The CoMTB induced IL-8 mRNA expression in the A549 cells was evaluated using RT-PCR, and CoMTB induced $I{\kappa}B{\alpha}$ degradation was measured using western blot analysis. CoMTB induced nuclear translocation and DNA binding of NF-${\kappa}B$ was also examined using an electrophoretic mobility shift assay(EMSA), and the CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity was measured using a luciferase reporter gene assay. Results : CoMTB induced IL-8 production by A549 cells($46.8{\pm}4.8\;ng/ml$) was higher than with direct stimulation with M. tuberculosis ($6.8{\pm}2.9\;ng/ml$). CoMTB induced IL-8 mRNA expression increased after 2 h of stimulation and was sustained for 24 h. $I{\kappa}B{\alpha}$ was degraded after 10 min of CoMTB stimulation and reappeared by 60 min. CoMTB stimulated the nuclear translocation and DNA binding of NF-${\kappa}B$. The CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity($13.6{\pm}4.3$ times control) was higher than either CoMCont($2.0{\pm}0.6$ times control) or M. tuberculosis ($1.4{\pm}0.6$ times control). Conclusion : A conditioned medium of peripheral blood monocytes phagocytosing M. tuberculosis stimulates NF-${\kappa}B$ dependent IL-8 production by the lung epithelial cells.

• Korean Journal of Clinical Laboratory Science
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• v.56 no.2
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• pp.115-124
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• 2024

The emergence and spread of Staphylococcus aureus, which is resistant to quinolone antibacterial agents, has made it difficult to treat infectious diseases. Accordingly, this study examined the molecular epidemiological characteristics of quinolone-resistant S. aureus (QRSA) to obtain helpful data for treatment. Mutations in mecA and SCCmec typing, gyrA, and gyrB genes were investigated for QRSA strains isolated from the blood culture specimens at a general hospital in Daejeon Metropolitan City. The ciprofloxacin-resistant strains in SCCmec typing were II (44 strains, 73%), IVa (five strains, 8%), III, and V (one strain, 2%); the non-typeable strains (11 strains, 18%), and levofloxacin (LVX) and moxifloxacin (MXF) strains were II (44 strains, 73%), IVa (five strains, 8%), III, and V (one strain, 2%); the non-typeable strains were 10 (17%). In both gyrA and gyrB regions, there were 58 mutations, or 96.7%. In LVX, there were 56 mutations or 93.3%, and in MXF, there were 57 mutations or 95%. Twelve mutations, six mutations each in gyrA and gyrB, were identified for the QRSA strain. The resistance rate for the quinolone antibiotics of QRSA studied was approximately 98%, and 12 mutations, six each in gyrA and gyrB, were identified in the QRSA strain. Therefore, the rational use of antibiotics needs to be improved.

• Animal Systematics, Evolution and Diversity
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• v.28 no.2
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• pp.133-136
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• 2012

The objective of this study was to determine the degree of mitochondrial DNA (mtDNA) divergence between two subspecies of $Mustela$ $sibirica$ from Korea ($M.$ $s.$ $coreanus$ on the Korean Peninsula and ($M.$ $s.$ $quelpartis$ on Jeju Island) and to examine the taxonomic status of ($M.$ $s.$ $quelpartis$. Thus, we obtained complete sequences of mtDNA cytochrome $b$ gene (1,140 bp) from the two subspecies, and these sequences were compared to a corresponding haplotype of ($M.$ $s.$ $coreanus$, downloaded from GenBank. From this analysis, it was observed that the sequences from monogenic ($M.$ $s.$ $quelpartis$ on Jeju Island were identical to the sequences of four ($M.$ $s.$ $coreanus$from four locations across the Korean Peninsula, and that the two subspecies formed a single clade; the average nucleotide distance between the two subspecies was 0.26% (range, 0.00 to 0.53%). We found that the subspecies $quelpartis$ is not genetically distinct from the subspecies $coreanus$, and that this cytochrome $b$ sequencing result does not support the current classification, distinguishing these two subspecies by pelage color. Further systematic analyses using morphometric characters and other DNA markers are necessary to confirm the taxonomic status of ($M.$ $s.$ $quelpartis$.

• Journal of Conservation Science
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• v.26 no.1
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• pp.95-107
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• 2010

Biochemical research was carried out on 4 human skeletal remains from historical lime-layered tombs assigned to the Joseon Dynasty in Oknam-ri, Seocheon. The preservation of femur was evaluated by stereoscopic microscopy and scanning electron microscopy. Most of specimens showed good histological preservation. The histological results proved to be a good potentiality for biochemical analysis using bio-molecules. The amelogenin gene and mitochondrial DNA (mtDNA) analyses revealed that three specimens perhaps have maternal consanguinity due to sharing with mtDNA haplogroup D4b1, and two specimens buried in the same tomb were a couple in Gatjaegol site. Carbon and nitrogen stable isotope analysis indicated that four deads diet were built around C3 plant as rice, barley, wheat and bean. In this study we characterized genetic and diet features from the social stratum who could make lime-layered tombs during period of the Joseon Dynasty. The results suggest that biochemical research using the human skeletal remains from the Joseon Dynasty has the great potential and reasonable value for archaeology, anthropology, and population genetics.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.749-754
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• 2000

PCR primers were designed based on consensus sequences of dTDP-D-glucose 4,6-dehydratase, one of the enzymes involved in the biosynthesis of deoxysugar. The PCR product (360 bp) was obtained from Thermus caldophilus GK24. Colony hybridization was carried out to the cosmid library constructed from T. caldophilus GK24 genomic DNA by the PCR product DNA fragment. We isolated a cosmid clone (pSMTC-1) that was subcloned to call pKCB series plasmid (BamHI fragments), partially sequenced and analyzed. pKCB80 (4.2 kb-BamHI DNA fragment) of them showed ORFs that was orfA, orfB, orfC and orfD. The orfABCD gene cluster is the deosysugar biosynthetic gene ; orfA (glucose-1-phosphate thymidylytransferase), orfB (dTDP-D-glucose 4,6-dehydratase), orfC (dTDP-4-keto-L-rhamnose reductase) and orfD (dTDP-4-keto-6-deoxy-D-glucose 3,5-epimerase). The gene cluster that was related in biosynthesis of dTDP-L-rhamnose was also identified by computer analysis, and we proposed that the biosynthetic pathway of deoxysugar analyzed from DNA sequencing of pKCB80 is from D-glucose-1-phosphate, dTDP-D-glucose, dTDP-4-keto-6-deoxy-D-glucose via dTDP-4-keto-L-rhamnose to dTDP-L-rhamnose.

• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.23-33
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• 1998

The entire sequence of a 4,214,810 bp genome of the Bacillus subtilis 168 has been determined by an international project, and the completion has been announced on July 19, 1997. For the sequencing project an international consortium was established and 25 European, 7 Japanese laboratories, 2 biotechnology companies, and our laboratory participated in the project. Within this framework we determined the complete nucleotide sequence of a 53,289 bp fragment upstream of the odhA gene (181 $^{\circ}$) of the B. subtilis 168 chromosome. On the basis of the published DNA sequences of the B. subtilis sspC and odhA genes, we obtained genomic fragments by plasmid rescue and long-range PCR. The sequenced fragment contains 56 putative open reading frames (designated yojA-yolI and 9 known genes (sspC, cge cluster, orfE5, orfRMl and odhA), in which we found many interesting features. In addition, the entire nucleotide sequence of a 53,289 bp region enabled us to revise the current genetic map of this region.

• Journal of Veterinary Clinics
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• v.26 no.5
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• pp.419-422
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• 2009

We performed the PARR (PCR to detect antigen receptor rearrangements) test on DNA isolated from twelve archival canine cytological slides including nine lymphoma, two reactive lymphocytes and one sample from Ehrlichia canis infected dog. As a result, our PCR control gene, $C{\mu}$, was successfully amplified from all of the DNA samples. Six out of nine lymphoma samples showed a clonal rearrangement of immunoglobulin gene whereas three samples did a clonal rearrangement of T cell receptor gamma ($TCR{\gamma}$) gene. However, we observed no visible or clear bands from PCR conducted using our antigen receptor rearrangement primers on DNA from a reactive lymphoid cell proliferation used as a negative control. False-positive amplification in $TCR{\gamma}$ gene was observed only in one sample from E. canis infection. The use of archival cytological specimens demonstrated in this study offers potential advantages for cost-effective specimen acquisition and efficient high-fidelity DNA analysis.

• Journal of Life Science
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• v.16 no.5
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• pp.812-827
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• 2006

To investigate the population structure and geographic distance among anchovies (Engraulis japonicus) in Korea, we compared and analyzed the mitochondrial DNA control region sequences (227 bp) of anchovies from 12 localities in inshore and offshore waters. The sequence analysis of 84 individuals showed 29 haplotypes, ranging in sequence divergence by pairwise comparisons from 0.3% to 3.5% (1 bp-12 bp). E9 haplotype of anchovies were found largely in inshore waters and also in offshore waters, which was regarded as the major source in Korean waters (58.3%). However, E26, E27, E28, and E29 haplotypes were found in westsouthern (locality 10, four among 7). Phylogenetic analysis using PHYLIP was divided into two clades (clade A and B). Most of the haplotypes, excluding E26, E27, E28, and E29, were strongly supported by bootstrap analysis (>75%), whereas the relationship between clade A and B was weakly supported by bootstrap analysis (51%). High levels of genetic diversity were found; haplotype diversity (H)=0.75-1.00, and nucleotide diversity $({\pi})=0.015-0.0244$. Analysis of $F_{ST}$ between populations in inshore waters ranged in 0.01-0.05, whereas those of offshore waters ranged in 0.01-0.58. A high gene flow occurred in inshore (Nm=22.61-34.22) and offshore (Nm=11.57-45.67) populations. The distribution of mitochondrial DNA haplotypes between westsouthern and other populations was suggestive of significantly different differentiation ($F_{ST}$=0.20-0.59, p<0.05; d=0.52, p=0.00; ${\phi}=0.02-0.41$, p＜0.05). These results suggested that the overall anchovy population in the Korean peninsula caused considerable migration due to the mitochondrial gene flow between inshore and offshore populations to form a genetically homogenous and panmictic structure, although a heterogeneous population was found in this study.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.298-304
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• 2007

Salmonella is facultative intracellular pathogen that can survive and replicate in macrophages even though these cells are equipped with a plethora of anti-microbial mechanisms. To survive in this hostile intracellular environment, Salmonella has evolved numerous defense mechartisms. An approximately 20 kDa protein was detected as a stationary-phase specific protein band in cytosolic fraction. It was identified as a DNA binding protein in stationary phase (Dps) by analysis of MALDI-TOF assay. It has been known that Dps, the protein produced in the stationary phase of bacteria, allows DNA to form chromatin by binding to DNA nonspecifically and protects DNA from reactive oxidative species (ROS). For further study, Dps specific polyclonal antibodies were generated by injection of purified Dps protein into rabbit. To examine the Finfluence of several regulatory proteins in the expression dps gene, Dps protein level in various S. typhimurium mutants defecting regulatory proteins were investigated by Westernblot using Dps specific polyclonal antibodies.