• Food Science and Biotechnology
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• v.15 no.2
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• pp.232-237
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• 2006

Bacillus cereus group comprising B. cereus, B. thuringiensis, and B. mycoides was differentiated by polymerase chain reaction (PCR) and colony morphology. Prevalence of B. cereus group in rice and distribution of enterotoxin genes were determined as possible food poisoning agents. PCR using primers targeted for gyrB and cry genes could distinguish B. thuringiensis from B. cereus, and B. mycoides was differentiated by rhizoid morphological characteristics on nutrient agar. Among 136 rice and their processed products, prevalence of B. cereus group was 40%. B. cereus group consisted of 54 B. cereus, 11 B. thuringiensis, and 1 B. mycoides. Major isolates were B. cereus, with B. thuringiensis detected up to 10% among edible rice tested. Five enterotoxin genes, hbl, nhe, bceT, entFM, and cytK, were broadly distributed among B. cereus group, especially in B. cereus and B. thuringiensis. Prevalence of B. cereus group in rice and enterotoxin distribution suggest B. thuringiensis and B. cereus are toxigenic strain that should be controlled in rice and its products.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.1
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• pp.124-130
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• 2008

Streptococcus (Strep.) agalactiae is one of the major pathogens of bovine mastitis and is the main cause of subclinical infection. This study attempted to develop a rapid PCR diagnosis procedure using milk samples collected on filter paper disks. Chromatographic filter paper was employed as the preservation media and kept at room temperature for one to four weeks. The revival rate of Strep. agalactiae kept on dried filter paper disks was affected by the pretreatment preservation time. The revival test suggested that not all the bacteria in artificially contaminated milk samples on the filter paper disks could be recovered. After that, a PCR based on the 16-23S intergenic spacer region of Strep agalactiae was performed. The results distinguished the strep. agalactiae from major pathogens of bovine mastitis at a $2{\times}10^2$ colony forming units (CFU)/ml level, which showed similar sensitivity to the results from liquid milk samples. The results also showed that milk samples collected on filter paper disks could be kept at room temperature for one to four weeks with little negative effect on sensitivity and specificity. The field test showed that the diagnostic sensitivity and specificity was 96.15% and 98.60%, respectively. In conclusion, the protocol will provide a rapid and economic procedure for the detection of bovine mastitis.

• Journal of Sensor Science and Technology
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• v.27 no.4
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• pp.237-241
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• 2018

The real-time enrichment and detection of pathogens are serious issues and rapidly evolving field of research because of the ability of these pathogens to cause infectious diseases. In general, bacterial detection is accomplished by conventional colony counting or by polymerase chain reaction (PCR) after DNA extraction. As colony counting requires considerable time to cultivate, PCR is an attractive method for rapid detection. A small number of pathogens can cause diseases. Hence, a pretreatment process, such as enrichment is essential for detecting bacteria in an actual environment. Thus, in this study, we developed a microfluidic chip capable of performing rapid enrichment of bacteria and the extraction of their genes. A lectin, i.e., Concanavalin A (ConA), which shows binding affinity to the surface of most bacteria, was coated on the surface of magnetic particles to nonspecifically capture bacteria. It was subsequently concentrated through magnetic forces in a microfluidic channel. To lyse the captured bacteria, magnetic particles were irradiated by a wavelength of 532nm. The photo-thermal effect on the particles was sufficient for extracting DNA, which was consequently utilized for the identification of bacteria. Our device will help monitor the existence of bacteria in various environmental situations such as water, air, and soil.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.1017-1021
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• 2013

The cellular apoptosis susceptibility (CSE1L) gene has been demonstrated to regulate multiple cellular mechanisms including the mitotic spindle check point as well as proliferation and apoptosis. However, the importance of CSE1L in human colon cancer is largely unknown. In the present study, we examined expression levels of CSE1L mRNA by semiquantitative RT-PCR. A lentivirus-mediated small interfering RNA (siRNA) was used to knock down CSE1L expression in the human colon cancer cell line RKO. Changes in CSE1L target gene expression were determined by RT-PCR. Cell proliferation was examined by a high content screening assay. In vitro tumorigenesis was measured by colony-formation assay. Cell cycle distribution and apoptosis were detected by flow cytometric analysis. We found CSE1L mRNA to be expressed in human colon cancer cells. Using a lentivirus based RNAi approach, CSE1L expression was significantly inhibited in RKO cells, causing cell cycle arrest in the G2/M and S phases and a delay in cell proliferation, as well as induction of apoptosis and an inhibition of colony growth capacity. Collectively, the results suggest that silencing of CSE1L may be a potential therapeutic approach for colon cancer.

• Korean Journal of Veterinary Research
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• v.44 no.4
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• pp.551-559
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• 2004

Shiga toxin-producing Escherichia coli (STEC) causes various clinical signs in human and animals, and has been indicated as a global enteropathogen with zoonotic importance. In this study, the feces of healthy pigs were collected from the slaughtered pigs of Daejon abattoir during the period from December 2001 to October 2002. Of 326 specimens, 13 STEC were confirmed by culture, PCR and colony hybridization. The isolates were further studied for toxin types, pathogenic factors, plasmid profiles, and antimicrobial resistance to characterize the genetic and toxigenic properties. In PCR, all of 13 isolates were evident to have shiga toxin gene (stx). Of 13 isolates stx1 gene was detected in 4 and stx2 gene in 9. The genes of eaeA, hlyA and rfbE were not present in any isolates. In colony hybridization using shiga toxin common primer (STXc), 2 to 9 per 100 colonies subcultured from 13 isolates showed the positive reaction. In the examination for plasmid profiles of the isolates, one to eleven plasmids with varying sizes of 1.0 Kb to 100 Kb were detected, and the 13 STEC could be classified into four groups by the plasmid patterns. The antimicrobial resistance patterns of the isolates were comparably corresponded with the plasmid profile patterns.

• Korean Journal of Soil Science and Fertilizer
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• v.44 no.6
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• pp.1181-1184
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• 2011

Total aerobic bacteria, Esherichia coli O157:H7, and Salmonella spp. were examined in commercial liquid pig manures. Commercial liquid pig manures (n=33) were collected from muck joint resource recovery plant at April, June, August, October 2009, Korea. Total aerobic bacteria were incubated at $37^{\circ}C$ for 24-48 hrs, and quantified as a colony-forming unit (CFU) $mL^{-1}$. Analysis of Esherichia coli O157:H7 and Salmonella spp. were followed by Korean Food Standards Codex method. Colony of Salmonella spp. was confirmed by API kit and real time polymerase chain reaction (PCR). Total aerobic bacteria isolated from fermented commercial liquid pig manures ranged from 2.8 to $24.3{\times}10^4\;CFU\;mL^{-1}$. Esherichia coli O157:H7 was not detected, and Salmonella spp. showed the low detection frequency at only 1 sample. This study suggests that continuous monitoring in commercial liquid pig manures is required to improve the agricultural food through management of agricultural land contaminated with liquid pig manures.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.52-52
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• 2003

Granulocyte-macrophage colony stimulating factor (GM-CSF) is synthesized in the female reproductive tract and has been shown to play an important role in human and murine embryo development and implantation. However, the mechanism of GM-CSF on the embryo development is unknown. Recent studies suggested that GM-CSF may be increase the expression of implantation relented genes, such as interleukin-1 (IL-1) system. Our aim of this study was to compare the interleukin-1$\alpha$ (IL-1$\alpha$), interleukin-1$\beta$ (IL-1$\beta$) and interleukin-1 receptor antagonist (IL-lra) mRNA between the GM-CSF supplemented group and control group in mouse embryos. Mouse 2-cell embryos were cultured in P-1 medium supplemented with or without mouse GM-CSF (10 ng/ml). The number of total and apoptotic cell in blastocyst were assessed by TUNEL. And then, the expression of IL-1$\alpha$, IL-1$\beta$ and IL-1ra mRNA in blastocyst were examined by RT-PCR.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.2
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• pp.633-636
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• 2012

Purpose: PRIL (proliferation-inducing ligand) is a newly identified member of the tumor necrosis factor (TNF) family and modulates death ligand-induced apoptosis. Here, we investigated the effect of PRIL on cellular characteristics relating to tumor progression in human gastric cancer. Method: Recombinant lentivirus containing PRIL siRNA was constructed and then infected MGC803 and SGC7901 gastric cancer cells. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] colony formation and cell cycle analysis were used to study the effect of PRIL knockdown on gastric cancer cell proliferation. Results: PRIL expression in lentivirus infected cells was significantly reduced as evidenced by quantitative real-time PCR. Cell viability and colony formation of MGC803 and SGC7901 cells were significantly hampered in PRIL knock-down cells. Moreover, the cell cycle was arrested at G2/M phase, elucidating the mechanism underlying the inhibitory effect of siRNA on cell proliferation. Conclusions: Our study indicated that PRIL functions in promoting cell growth, and lentivirus-mediated PRIL gene knockdown might be a promising strategy in the treatment of gastric cancer.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.1
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• pp.35-43
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• 2011

Thirty five isolates of Trichoderma species collected from seven different locations of Bangladesh were studied for morphological characters and molecular variation. Mycelial diameters of the isolates varied from 8.28 cm to 9.00 cm. Based on colony colour, isolates were grouped into five such as dark green, green, light green, yellowish green and whitish green. Maximum isolates were green and light green. On the basis of growth habit and colony consistency, the isolates were categorized into three groups, in which most species had fast growth and were compact in appearance. PCR-based Random Amplified Polymorphic DNA (RAPD) technique employing 3 decamer primers produced 36 scorable bands of which all (100%) were polymorphic. The co-efficient of gene differentiation (Gst) was 1.0000 reflecting the existence of high level of genetic diversity among the isolates. The Unweighted Pair Group Method of Arithmetic Means (UPGMA) dendrogram constructed from Nei's (1972) genetic distance produced 2 main clusters (13 isolates in cluster 1 and 22 isolates in cluster 2). The result indicating their genetic diversity has opened new possibility of using the most efficient and more isolates of Trichoderma in the preparation of biopesticide and decomposition of municipality waste.

• Microbiology and Biotechnology Letters
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• v.31 no.1
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• pp.18-24
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• 2003

For researching nitrogen-fixing bacteria associated with gramineous crops, we collected growing roots of rices, wheats, oats, barleys, ryes, and maizes at 19 sites of southern Korean peninsula. Endophytes and free living bacteria were isolated from those crop roots. Sixty-three isolates were classified on the basis of different morphology, size, color, host of colony, and the 16S rDNAs sequence. The analyses of PCR amplification for nifH gene and nitrogenase activity assay, revealed that all isolates contained nitrogen-fixing abilities. In addition, most of them have cellulase activity which is one of the common features of endophytic bacteria from plant.