• Asian Pacific Journal of Cancer Prevention
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• v.14 no.1
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• pp.201-208
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• 2013

Objective: To investigate changes in the invasive capacity of gastric cancer cells in vitro after expression inhibition of T lymphoma invasion and metastasis inducing factor 1 (Tiam 1) and underlying mechanisms. Methods: Using adhesion selection, two subpopulations with high ($M_H$) or low ($M_L$) invasive capacity were separated from the human gastric cancer cell line MKN-45 ($M_0$). Tiam 1 antisense oligodeoxynucleotide (ASODN) was transfected into $M_H$ cells with liposomes, and expression of Tiam 1 mRNA and protein was determined by RT-PCR and quantitative cellular-ELISA. Changes in the cytoskeleton, invasive capacity in vitro and expression of ras-related $C_3$ botulinum toxin substrate 1 (Rac 1), integrin ${\beta}1$ and matrix metalloproteinase 2 (MMP 2) between Tiam 1 ASODN transfected $M_H$ cells and non-transfected cells were observed by HE staining, cytoskeletal protein staining, scanning electron microscopy, Boyden chamber tests and cyto-immunohistochemistry. Results: A positive correlation existed between the expression level of Tiam l mRNA or protein and the invasion capacity of gastric cancer cells. After ASODN treatment ($0.43{\mu}M$ for 48 h), Tiam 1 mRNA transcription and protein expression in $M_H$ cells were decreased by 80% and 24% respectively (P < 0.05), compared with untreated controls, while invasive capacity in vitro was suppressed by 60% (P < 0.05). Morphologic and ultrastructural observation also showed that ASODN-treated $M_H$ cells exhibited smooth surfaces with obviously reduced filopodia and microspikes, which resembled $M_0$ and $M_L$ cells. Additionally, cytoskeletal distribution dramatically altered from disorder to regularity with reduced long filament-like structure, projections, pseudopodia on cell surface, and with decreased acitn-bodies in cytoplasm. After Tiam 1 ASODN treatment, the expression of Rac 1 and Integrin ${\beta}1$ in $M_H$ cells was not affected (P > 0.05), but that of MMP 2 in $M_H$ cells was significantly inhibited compared with untreated cells (P < 0.05). Conclusion: Over-expression of Tiam-1 contributes to the invasive phenotype of gastric cancer cells. Inhibition of Tiam 1 expression could impair the invasive capacity of gastric cancer cells through modulating reconstruction of the cytoskeleton and regulating expression of MMP 2.

• Korean Journal of Microbiology
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• v.55 no.1
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• pp.74-76
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• 2019

Gram-positive anaerobic bacilli Actinomyces spp. commonly reside on mucosal surfaces of the oropharynx, gastrointestinal tract, and urogenital tract. Here, we first report the draft genome sequence of Actinomyces georgiae KHUD_A1, isolated from dental plaque of a Korean elderly woman. The genome is 2,652,059 bp in length and has a GC content of 68.06%. The genome includes 2,242 protein-coding genes, 9 rRNAs, and 64 tRNA. We identified 157 KHUD_A1 strain-specific genes, including genes encoding CPBP family intramembrane metalloprotease, bile acid: sodium symporter family protein, Txe/YoeB family addiction module toxin and Phd/YefM family antitoxin. The sequence information of A. georgiae KHUD_A1 will help understand the general characteristics of the bacterial species and the genome diversity of the genus Actinomyces.

• Journal of Food Hygiene and Safety
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• v.16 no.3
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• pp.232-240
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• 2001

One hundred twenty five bacterial isolates were obtained from the brown blotch-diseased oyster mushrooms collected from markets. Among them, 45 were determined as pathogenic bacteria and white line forming organisms(WLFO) were 6 strains and white line reaction organisms (WLRO) were 6 strains. All of the white line forming isolates were identified as Pseudomonas tolaasii which is a known pathogen of brown blotch disease of oyster mushroom by GC-MIS(Gas chromatography-microbial identification system). Six of the white line reacting organisms were identified as P. chlomraphis, P. fluorescens biotype A and type C. The rest of them were P gingeri, P. agarici, P. fluorescens biotype B, P. chloroyaphis, non-pathogenic P. tolaasii, P. putida biotype A and B etc. For spectrum of activity of tolaasin, culture filtrates from pathogenic isolates were examined by browning of mushroom tissue and pitting of mushroom caps. The weak pathogenic bacteria didn't induce browning or pitting of mushroom tissue. On the other hand, strong pathogenic isolates showed browning and pitting reaction on mushroom. An extracellular toxin produced by P. tolaasii, was investigated. The hemolysis activity test of 6 strains identified as P. tolaasii were 0.8∼0.9 at 600 nm and 3 strains of WLRO were 0.9∼1.0 and Pseudomonas app. were 1.0∼1.2. Observation of fresh mushroom tissue using confocal laser scanning microscopy was carried out for images of optical sectioning and vertical sectioning. Also images of brown blotch diseased oyster mushroom tissue after contamination P. tolaasii was obtained by CLSM.

• KSBB Journal
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• v.18 no.6
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• pp.522-528
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• 2003

The purpose of this study is to inquire into molecular epidemiological characteristics of Vibrio parahaemolyticus. For this study, 120 strains(120 strains of Vibrio parahaemolyticus sampled from diarrhea patients) were examined and analyzed for biochemical characteristics, TDH (thermostable direct hemolysin) antibiotics sensitivity and detection of toxR, gyrE, tdh, and tds gents. G-S PCR (Group Specific Polymerase), PFGE (Pulsed-field Gel Electrophoriesis) methods were performed on the materials from patients were results. 1 Vibrio parahaemolyticus didn't grow in 0% density of NaCl, but the fact was found that those grew in 8% density of NaCl. 2. O：K serotypes of Vibrio parahaemolyticus was turned up in domestic patients was 17 types. Among those O3：K6 was the most, it was 68.3%. 3. In 18 kinds of antibiotic tests resistant against Ampicillin, Ticacillin was comparatively high. the case of resistant against Ampicillin, Ticacillin, Vancomycin at the multiple resistant was 52.5%. 4. Toxin gene tdh had only 109 strains among 120 ones isolated from patients held the genes of 199bp size, and 11 strains was negativity 5. In the test of Kanagawa toxic productivity, 107 strains among strains isolated from patients appeared to be positivity reaction 6. The strain that held trh toxin was only 3, and those among test strains had the genes of 250bp size and that had tdh, trh genes at a time were 3 strains, and TDH toxic productivity of those were 16 times, and it was weak. 7. Group Specific-PCR appeared to be useful in the confirmation of O3:K6 serotype interrelations. 8. Three strains which showed difference of 7 DNA sequence even in the same serotype were detected by the result of analyzing the regular gene, toxRS DNA sequence. These strains are differ from general strains which carry infection easily. 9. These mutual dose epidemiological relations were classified into smaller-parts through PFGE method. As a result of such classify, 3 findings were found. V. parahaemolyticus sampled from diarrhea patients were classified into 3 types. And third, the result obtained through PFGE method can be used as a useful tool in a point of molecular-epidemiological view.

• Development and Reproduction
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• v.11 no.1
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• pp.49-54
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• 2007

Ethane 1,2-dimethane sulfonate(EDS), a toxin which specifically kills Leydig cells(LC), has been widely used to prepare the reversible testosterone(T) depletion rat model. In the present study, we monitored the gene expression profiles of pituitary gonadotropins, LH and FSH, up to 7 weeks after EDS injection. Adult male Sprague-Dawley rats($300{\sim}350\;g$ B.W.) were injected with a single dose of EDS(75 mg/kg i.p.) and sacrificed on weeks 0, 1, 2, 3, 4, 5, 6 and 7. Total RNAs were purified from each pituitary, and the message levels of common alpha subunit($C{\alpha}$) of pituitary glycoprotein hormones, LH beta subunit($LH{\beta}$), FSH beta subunit($FSH{\beta}$) and GnRH receptor(GnRH-R) were evaluated by semi-quantitative RT-PCRs. The message levels of $C{\alpha}$ increased sharply during weeks 1-4, then return to the control level on week 5. The mRNA levels of $LH{\beta}$ were elevated after week 2, reached the peak at week 4, then declined to the control level after week 5. The message levels of $FSH{\beta}$ were elevated after week 2, reached the peak at week 3, then declined to the nadir at week 5. Similarly, the mRNA levels of GnRH-R were elevated after week 2, reached the peak at week 3, then gradually declined to the control level after week 5. The present study indicated that EDS treatment could induce reversible alterations in the transcriptional activities of gonadotropin subunits and GnRH-R in the anterior pituitary from male rats. EDS injection model might be useful to understand the mechanism of hormonal regulation of hypothalamus- pituitary neuroendocrine axis in male rats.

• Research in Plant Disease
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• v.23 no.3
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• pp.278-282
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• 2017

Maize is one of the most cultivated cereals as a staple food in the world. The harvested maize is mainly stored after drying, but its quality and nutrition could be debased by fungal spoilage and mycotoxin contamination. In this study, we surveyed mycotoxin contamination fungal infection of maize kernels that were stored for almost one year after harvest in 2015. The amount of deoxynivalenol and zearalenone detected were higher than the other mycotoxin, such as aflatoxin, ochratoxin, fumonisin and T-2 toxin. In particular, level of deoxynivalenol was detected as $1200{\pm}610{\mu}g/kg$ in small size kernels, which was four to six times higher than the large and the medium size kernels. Moreover, the amount of deoxynivalenol, zearalenone, and fumonisin were increased with discolored kernels. 10 species including Fusarium spp., Aspergillus spp. and Penicillium spp. were isolated from the maize kernels. F. graminearum was predominant in the discolored kernels with detection rates of 60% (red) and 40% (brown). Our study shows that the mycotoxin contents of stored maize can be increased by discolored maize kernels mixed. Therefore elimination of the contaminated maize kernels will help prevent fungal infection and mycotoxin contamination in stored maize.

• Journal of Plant Biotechnology
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• v.40 no.1
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• pp.18-26
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• 2013

Genetically modified (GM) rice Agb0101, which expresses the insecticidal toxin modified cry1Ac (mcry1Ac1) gene, was developed by the Rural Development Administration in Korea. To monitor the probable release of Agb0101 in the future, it is necessary to develop a reliable detection method. Here, we developed the PCR detection method for monitoring and tracing of GM rice. The primer pair (RBEgh-1/-2) from a starch branching enzyme (RBE4) gene was designed as an endogenous reference, giving rise to an expected PCR amplicon of 101 bp. For the qualitative PCR detection, construct- and event-specific primers were designed on the basis of integration sequence of T-DNA. Event-specific PCRs amplified specifically 5'- or 3'-junction region spanning the native genome DNA and the integrated gene construct, while none of amplified product was shown on crops, rice varieties, and other insect-resistant transgenic rice lines. The event-specific real-time PCR method was performed using TaqMan probe and plasmid pRBECrR containing both rice endogenous gene RBE4 sequence and 5'-junction sequence as the reference molecule. The absolute limit of quantification (LOQ) of real-time PCR was established with around 10 copies for one plasmid molecule pRBECrR. Thereafter, the different amounts of transgenic rice (1, 3, 5, and 10%, respectively) were quantified by using the established real-time PCR method, with a range below 19.55% of the accuracy expressed as bias, 0.06-0.40 of standard deviation (SD) and 3.80-7.01% of relative standard deviations (RSD), respectively. These results indicate that the qualitative and quantitative PCR methods could be used effectively to detect the event Agb0101 in monitoring and traceability.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.5
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• pp.612-617
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• 2008

Tetrodotoxin (TTX) analogues were first determined from the liver extracts of puffer fish, Takifugu niphobles, by LC/MS with Hydrophilic Interaction Liquid Chromatography (HILIC). In total, 7 TTX analogues were detected within 20 minutes as follows; 5,6,11-trideoxyTTX (34.0%, 1,029.6 nmol/g), 6,11-dideoxyTTX (29.3%, 887.6 nmol/g), TTX (22.1%, 667.8 nmol/g), 4,9-anhydro-TTX (11.2%, 339.3 nmol/g), 11-deoxyTTX＋5-deoxyTTX (2.6%, 78.6 nmol/g), and 4-epiTTX (0.8%, 23.6 nmol/g). Mouse toxicity of diluted liver extracts showed the highest toxicity at pH 3 (8.7 MU/mL) and decreased, as increasing pH, to 1.4 MU/mL at pH 10. At acidic (pH 5) and neutral conditions (pH 7), mouse toxicity of liver extracts (79 MU/mL) decreased slowly, as increasing temperature from $80^{\circ}C$ to $115^{\circ}C$, and time until 1 hour; in contrast, at the akaline condition (pH 9), the toxicity decreased rapidly to the more than half within 10 minutes. Individual toxicity of the fillet of T. niphobles were between $43.2{\sim}106.7$ MU, and $64{\sim}78%$ of its toxicity was eluted to soup when boiled with 3 volumes of water during 10 minutes.

• Korean Journal of Animal Reproduction
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• v.24 no.3
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• pp.241-248
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• 2000

In this study, we examined the number and motility of sperms, and also observed the changes in testes weight, and histological changes of several organs after 5 days of a continuous administration of dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin) of per oral administration of a herb drug extracts, which were administerated altermate days, to elucidate the effects of the a herb drug extracts on reproductive toxicity of dioxin. 1. The sperm numbers of dioxin-administered groups were 90.7$\pm$3.6~l18.5$\pm$3.6$\times$10/suup 6/$m\ell$ and 67.3$\pm$4.1~88.2$\pm$3.3$\times$10$^{6}$ $m\ell$ for 10~20 $\mu\textrm{g}$/kg and 30~40 $\mu\textrm{g}$/kg dosages of dioxin-administerated groups, respectively. Each dioxin-administered group showed prominently lower value than that of control group's which was 119.3$\pm$3.4~120.2$\pm$4.7 $\times$ 10$^{6.}$$m\ell$. 2. The sperm motility of each dioxin-administered group's also showed lower value than that of control group's which was 93.6$\pm$3.8~94.9$\pm$3.4%. The sperm motility of each dioxin-administerated group were 77.0$\pm$4.7~89.5$\pm$3.6% and 66.5$\pm$3.3 ~79.9$\pm$3.8% for 10~20 $\mu\textrm{g}$/kg and 30~40 $\mu\textrm{g}$/kg dosages of dioxin-administerated groups, respectively. 3. The sperm numbers of each group, which was administered a herb drug extracts, were 77.4$\pm$3.2~90.9$\pm$3.4$\times$10$^{6}$ $m\ell$, 78.0$\pm$3.3~105.0$\pm$4.2$\times$10$^{6}$ $m\ell$, 76.2$\pm$2.8~84.4$\pm$3.5$\times$10$^{6}$ $m\ell$ and 75.4$\pm$3.3~80.2$\pm$3.3$\times$10$^{6}$ $m\ell$ for extracts of green leaf, red ginseng, Kugija and Oume-administered groups respectively. And the sperm motility of each group were 63.4$\pm$3.8~77.0$\pm$4.0%, 65.5$\pm$4.1~87.4$\pm$3.8%, 64.3$\pm$4.2~69.8$\pm$4.2%, 66.3$\pm$3.9~66.0$\pm$4.0% for extracts of green leaf, red ginseng, Kugija and Oume extracts-administered groups, respectively. 4. The number and motility of sperm of control group were 119.3$\pm$3.4~120.2$\pm$4.7$\times$10$^{6}$ $m\ell$ and 93.0$\pm$3.5~96.1$\pm$3.5%, respectively. Red ginseng extracts- administered group seemed to be recovered than my other groups, and the green leaf extracts-administered group was shown to be the next. The Kugija and Oume extracts-administered groups didn't show to be recovered much. 5. Most a herb drug extracts-administered group except the red ginseng-administered group displayed prominently lower values of testes weights than that of control group's which was 0.15$\pm$0.01~0.16$\pm$0.01g. The red ginseng extracts-administered group seemed to be recovered conspicuously. 6. After 5 days of a continuous administration of 30 $\mu\textrm{g}$/kg dosage of dioxin followed by 3 weeks of per oral administration of green leaf, red ginseng, Kugija, or Oume extracts, histological findings showed that the liver, spleen, and testis of most a herb drug extracts-administered groups were damaged severely. By the way, the testes of red ginseng extracts-administered group seemed to recover compared to the other group's.ompared to the other group's.