• The Plant Pathology Journal
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• v.35 no.1
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• pp.51-62
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• 2019

A great variable response was observed when PP-3 and PP-J encumbered with 116 populations of root knot nematode (RKN) at two different temperatures ($25{\pm}2^{\circ}C$ and $30{\pm}2^{\circ}C$) and concentrations ($10^4$ and $10^5$ spores/ml). The PCR reaction amplified intergenic region between cytochrome oxidase subunit II gene (COII) and large subunit of rRNA gene (lrRNA) of the mitochondrial genome of different RKN species. The primer C2F3 and 1108 identified M. incognita with the highest frequency (52.6%) followed by M. javanica (36.8%) and M. arenaria (10.5%). The sizes of PCR products were 1.7 kb for M. incognita and M. javanica populations while populations of M. arenaria produced 1.1 kb fragment. The digestion with Hinf I yielded three different fragment length patterns on 1.5 % agarose gel. From current research it is concluded that intra-Meloidogyne genetic variability exist in RKN populations which have better encumbrance with P. penetrans.

• BMB Reports
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• v.38 no.5
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• pp.609-618
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• 2005

$\gamma$-Glutamyl transpeptidase (GGT, EC 2.3.2.2.) catalyzes the transfer of the $\gamma$-glutamyl moiety from $\gamma$-glutamyl containing ompounds, notably glutathione (GSH), to acceptor amino acids and peptides. A second gene (GGTII) encoding GGT was previously isolated and characterized from the fission yeast Schizosaccharomyces pombe. In the present work, the GGTII-lacZ fusion gene was constructed and used to study the transcriptional regulation of the S. pombe GGTII gene. The synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene was significantly enhanced by NO-generating SNP and hydrogen peroxide in the wild type yeast cells. The GGTII mRNA level was increased in the wild-type S. pombe cells treated with SNP. However, the induction by SNP was abolished in the Pap1-negative S. pombe cells, implying that the induction by SNP of GGTII is mediated by Pap1. Fermentable carbon sources, such as glucose (at low concentrations), lactose and sucrose, as a sole carbon source, enhanced the synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene in wild type KP1 cells but not in Pap1-negative cells. Glycerol, a non-fermentable carbon source, was also able to induce the synthesis of $\beta$-galactosidase from the fusion gene, but other non-fermentable carbon sources such as acetate and ethanol were not. Transcriptional induction of the GGTII gene by fermentable carbon sources was also confirmed by increased GGTII mRNA levels in the yeast cells grown with them. Nitrogen starvation was also able to induce the synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene in a Pap1-dependent manner. On the basis of the results, it is concluded that the S. pombe GGTII gene is regulated by oxidative and metabolic stress.

• Tuberculosis and Respiratory Diseases
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• v.41 no.5
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• pp.462-474
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• 1994

Background : In acute lung injury, activated neutrophils play an important role in tissue damage. For neutrophils to participate in lung inflammation, chemotactic factors released from mononuclear phagocytes are needed to bring these cells to the local site of inflammation, with interleukin-8 (IL-8) being one of the most specific and important chemotactic factors for neutrophils. IL-8 also induces the expression of adhesion molecules and activates neutrophils to release various inflammatory mediators. Lipopolysaccharide(LPS) is one of the most important causes of adult respiratory distress syndrome and can cause release of many inflammatory cytokines including IL-8 leading to acute lung injury. But little is known about the regulatory mechanism of LPS-induced IL-8 gene expression in mononuclear phagocytes. Method : Human alveolar macrophages(HAM) and peripleral blood monocytes(PBMC) were isolated from healthy volunteers. Time and dose relationship of LPS-induced IL-8 mRNA expression was observed by Northern blot analysis. To evaluate the regulatory mechanism of LPS-induced IL-8 gene expression, pretreatment of actinomycin D(AD, $5{\mu}g/ml$) and cycloheximide(CHX, $5{\mu}g/ml$) was done and Northern blot analysis for IL-8 mRNA and ELISA for immunoreactive IL-8 protein in culture supernatant were performed. Results : 1) In HAM, dose and time dependent LPS-induced IL-8 mRNA expression was observed with peak mRNA level at 8 hours post-stimulation. 2) In PBMC, dose and time dependent LPS-induced IL-8 mRNA expression was also observed with peak mRNA level at 4 hours post-stimulation. 3) AD decreased expression of LPS-induced IL-8 gene expression at both mRNAand protein levels in both types of cells. 4) CHX decreased expression of LPS-induced IL-8 gene expression at protein level in both cell types but in HAM, superinduction of IL-8 mRNA was observed while decreased expression of IL-8 mRNA was observed in PBMC. Conclusion : Time and dose dependent LPS-induced IL-8 gene expression was observed in mononuclear phagocytes which is at least partly regulated pretranslationally. LPS-induced IL-8 mRNA expression in HAM needs no de novo protein synthesis and may be under the control of a labile repressor protein while de novo protein synthesis may be needed in PBMC.

• Reproductive and Developmental Biology
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• v.34 no.2
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• pp.103-109
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• 2010

Functional regulation of a specific tissue or organ is controlled by a number of ways, including local cell-cell interaction. Of several forms of cell-cell junctional complexes, gap junctions are caught a great attention due to a formation of direct linkage between neighboring cells. Gap junctions are consisted of connexin (Cx) isoforms. In the present study, we evaluated expressional profiling of Cx isoforms in the rat initial segment (IS) of the male reproductive tract at different postnatal ages. The presence and expression of 13 Cx isoform mRNAs were determined by semi-quantitative real-time PCR analyses. A total of 8 Cx isoform mRNAs were detected in the IS of the male rats during postnatal development. The highest level of Cx30.3 mRNA was found at 5 months of age, while abundance of Cx31 mRNA was the highest at 1 year of age. Expression of Cx31.1 gene was relatively consistent during the postnatal development. Fluctuation of Cx32 and 37 gene expression was observed during the postnatal period. Significant elevation of Cx40 mRNA abundance was detected at 25 days of age and older ages. Expression patterns of Cx43 and 45 genes were similar with the highest level at 2 weeks of age, followed by gradual decreases at older ages. These results indicate differential regulation on expression of Cx isoforms in the rat IS during postnatal development. A complicated regulation of gene expression of Cx isoforms in the IS at different postnatal ages is suggested.

• Biomedical Science Letters
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• v.10 no.2
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• pp.65-73
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• 2004

I report that single-stranded antisense as a part of large circular (LC-) genomic DNA of recombinant M13 phage exhibits enhanced stability, sequence specific antisense activity, and no need for target site search. A cDNA fragment (708 bp) of rat TNF-$\alpha$ was inserted into a phagemid vector, and TNF-$\alpha$ antisense molecules (TNF$\alpha$-LCAS) were produced as single-stranded circular DNA. When introduced into a rat monocyte/macrophage cell line, WRT7/P2, TNF$\alpha$-LCAS was able to ablate LPS-induced TNF-$\alpha$ mRNA to completion. The antisense effect of TNF$\alpha$-LCAS was shown to be sequence-specific because expressions of three control genes ($\beta$-actin, GAPDH and IL-1$\beta$) were not significantly altered by the antisense treatment. Further, TNF$\alpha$-LCAS was found to be highly efficacious as only 0.1 $\mu$g (0.24 nM) of TNF$\alpha$-LCAS was sufficient to block TNF-$\alpha$ expression in 1$\times10^5$ WRT7/P2 cells. I have also observed specific antisense activity in reduction of NF-$\kappa$B gene expression. The results suggest that an antisense sequence as a part of single-stranded circular genomic DNA has a specific antisense activity.

• Korean Journal of Soil Science and Fertilizer
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• v.44 no.1
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• pp.53-57
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• 2011

The MuSI is known as a multiple stress resistant gene with several lines. A previous study using RT-PCR showed that the expression of MuSI gene in tobacco plant induced its tolerance to Cd stress. This study was conducted to examine the enhanced Cd tolerance of the MuSI transgenic tobacco plant through germination test and to understand the role of the involved antioxidant enzymes for the exhibited tolerance. Germination rate of MuSI transgenic tobacco was more than 10% higher than that of wild-type tobacco, and seedlings of MuSI transgenic tobacco grew up to 1.6 times larger and greener than seedlings of wild-type tobacco at 200 and 300 ${\mu}M$ Cd. From the third to the fifth day, CAT activities at 100 and 200 ${\mu}M$ Cd and APX activities at 100, 200 and 300 ${\mu}M$ Cd of MuSI transgenic tobacco were up to two times higher than those of wild-type tobacco. MuSI gene is shown to enhance the activities of antioxidant enzymes resulting in higher tolerance to oxidative stress compared with the control plant.

• Journal of Life Science
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• v.17 no.10
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• pp.1426-1433
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• 2007

To investigate the antibiotic resistant patterns of the bacteria producing ESBL, we isolated one organism of Klebsiella pneumoniae from a clinical laboratory in Busan. The organism that produces ESBL gene was detected by double disk synergy test and the presence of three ESBL genes (TEM-1, SHV-12, CTX-M-15) was confirmed by polymerase chain reaction and DNA sequencing analysis. To analyse the characteristics of three ESBL genes, we performed transconjugation, transformation and cloning experiment with the organism. The MIC of Klebsiella pneumoniae was revealed that ceftazidime, cefotaxime and ceftriaxone were $256\;{\mu}g/ml,\;128\;{\mu}g/ml\;and\;128\;{\mu}g/ml$ respectively. The MIC of conjugant (E. coli $RG176^{Na(r)}$) af was revealed that ceftazidime, cefotaxime and ceftriaxone were $256\;{\mu}g/ml,\;64\;{\mu}g/ml\;and\;128\;{\mu}g/ml$ respectively. The MIC of transformant (E. cofi $DH5{\alpha}$) was revealed that ceftazidime, cefotaxime and ceftriaxone were $128\;{\mu}g/ml,\;32\;{\mu}g/ml,\;and\;32\;{\mu}g/ml$ respectively, The MIC of cloned organism of SHV-12 gene (E. coli $DH5{\alpha}$) was revealed that ceftazidime, cefotaxime and ceftriaxone were $128\;{\mu}g/ml,\;8\;{\mu}g/ml,\;and\;32\;{\mu}g/ml$ respectively. The results indicated that MIC of conjugant was higher than MIC of transformant and also SHV-12 gene were not resistant against cefotaxime antibiotic.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.16
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• pp.6613-6618
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• 2014

Background: The X-ray repair cross-complementing group 3 (XRCC3) is a highly suspected candidate gene for cancer susceptibility. Attention has been drawn upon associations of the XRCC3 Thr241Met polymorphism with breast cancer risk. However, the previous published findings remain controversial. Hence, we performed a meta-analysis to accurately evaluate any association between breast cancer and XRCC3 T241M (23, 812 cases and 25, 349 controls) in different inheritance models. Materials and Methods: PubMed and Web of Science databases were searched systematically until December 31, 2013 to obtain all the records evaluating the association between the XRCC3 Thr241Met polymorphism and breast cancer risk. Crude odds ratios (ORs) together with 95% confidence intervals (CIs) were used to assess the strength of associations. Results: When all eligible studies were pooled into the meta analysis of XRCC3 T241M polymorphism, a significantly increased breast cancer risk was observed in heterozygote comparison (OR=1.06, 95%CI=1.01-1.12). No significant associations were found in other models. In subgroup analysis, this polymorphism seemed to be associated with elevated breast risk in Asians. No publication bias was detected. Conclusions: This meta-analysis suggests that the T241M polymorphism confers a weakly increased breast cancer risk. A study with the larger sample size is needed to further evaluate gene-gene and gene-environment interactions of the XRCC3 T241M polymorphism with breast cancer risk.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.87-94
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• 2008

Porcine epidemic diarrhea virus (PEDV) is an infectious and highly contagious virus of swine. In order to develop the transgenic tobacco culture cells producing PEDV antigen protein, four vectors expressing PEDV spike protein (SP) gene under the control of a CaMV 35S promoter were constructed. Four fragments of the SP region of PEDV, SP1 (444 bp, 1487-1930 bp), SP2 (1.7 kb, 2300-3987 bp), SP3 (1.4 kb, 1559-2950 bp), and SP4 (2.6 kb, 9-2643 bp) were amplified by PCR and then C-MYC tag was fused to the end of each SP gene, respectively. These cassettes are inserted into the pCAMBIA2300 (named as 35S::SP1-M, 35S::SP2-M 35S::SP3-M, and 35S::SP4-M, respectively). Tobacco (cv. BY-2) cultured cells were transformed by co-cultivation with Agrobacterium tumefaciens harboring expression vector. We selected kanamycin-resistant calli and checked for the presence of the introduced SP gene using PCR, resulting 70% of them showed the foreign gene. We selected the lines with high-level expression of PEDV antigen protein based on dot blot analysis. Southern blot analysis confirmed that the PEDV SP gene was integrated into the genome of the tobacco cultured cells. Northern blot analysis showed that the introduced gene was highly expressed in transgenic cultured cells. Transgenic tobacco cultured cells-derived antigen induced immunogenicity in mice as determined by a plaque reduction neutralization assay. These results suggest that the vectors expressing PEDV spike protein gene in this study will be useful for the development of transgenic plants and cultured cells producing PEDV antigene protein.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.15
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• pp.6445-6449
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• 2015

Background: NSCLC is a disease involving uncontrolled cell growth, which could result in metastases into nearby tissues beyond the lungs. Materials and Methods: The aim of the present study was to analyze the influence of epidermal growth factor receptor (EGFR) gene expression on metastasis and survival in NSCLC patients. The present case-control study included 100 cases of NSCLC patients and 100 age and sex matched controls. EGFR gene expression was analyzed by quantitative real time PCR using serum RNA. Association with NSCLC patient survival was analyzed by the Kaplan-Meier method. Results: We analyzed EGFR gene expression and observed mean increased gene expression of 13.5 fold in NSCLC patients. Values reflected overall survival of patients with a median of 15.8 months in the cases of <13 fold increased gene expression vs 6.7 months with >13 fold increased EGFR gene expression (p=0.005). Distant metastatic patients with <13 fold increased EGFR gene expression had 7.9 months of median survival time while>13 fold increased EGFR gene expression had only 5 months of median survival time (p=0.03). Non metastatic patients with <13 fold increased EGFR gene expression had 18 months of median survival time as compared to only 7.1 months with >13 fold increased expression. Conclusions: Higher cell free EGFR mRNA expression may play an important role in causing distant metastases and reducing overall survival of NSCLC patients in the Indian population.