• Title/Summary/Keyword: IS-PCR

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Assessment Report of Bacterial Contamination in Some School Dining Services with Table Swabs and Air Samples (학교식당 및 교실배식 과정 전·후 미생물 오염에 관한 연구)

  • Jung, HeaYong;Sohn, JuHae;Lee, JaeYoon;Lee, InAe;Ko, JiYean;Ko, NaYun;Park, SungJun;Ko, GwangPyo;Kim, Sungkyoon
    • Journal of Environmental Health Sciences
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    • v.41 no.6
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    • pp.397-404
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    • 2015
  • Objectives: The aim of this study is to investigate microbial contamination in the school food service environment for the assessment of microbial food safety. Methods: We collected both swab samples from tables and desks and airborne bacterial samples from an elementary school (School A) and a high school (School B). Heterotrophic plate count, total coliform, Staphylococcus aureus, and Bacillus cereus were measured with selective media to quantify microbial concentration. PCR assay targeting 16S rRNA genes was performed to identify the strains of S. aureus and B. cereus isolated. In addition, we made a food service checklist for the locations to evaluate the food service environment. A Wilcoxon test was employed to examine the differences in microbial concentration between before lunchtime and afterwards. Results: Heterotrophic plate counts showed higher levels after-lunch compared to before-lunch at School B. However, levels of S. aureus were higher in the after-lunch period (p<0.05) in both classrooms and in the cafeteria in School A. B. cereus was only sparsely detected in School B. Several samples from food dining carts were found to be contaminated with bacteria, and facilities associated with food delivery were found to be vulnerable to bacterial contamination. Although microbial concentrations in the air showed little difference between before- and after-lunchtime in the cafeteria in School A, those in classrooms were greater after-lunchtime at both schools. Conclusion: Our results suggested that the microbial safety in schools after lunchtime of concern. Necessary preventive measures such as hygiene education for students and food handlers should be required to minimize microbial contamination during food service processes in schools.

Molecular Characterization of Escherichia coli Isolates from Humans and Chickens in the Chungcheong Area Using MLST Analysis (충청지역에서 분리된 사람 유래 대장균 및 닭 유래 대장균의 항균제 내성 및 MLST를 이용한 유전형의 분포 조사)

  • Kim, Semi;Sung, Ji Youn;Choi, Seung-Gu
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.2
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    • pp.71-77
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    • 2015
  • Antimicrobial resistant bacteria has recently emerged and been disseminated in livestock environments because of excessive use of antimicrobial agents for the therapeutic and growth promotion purposes to food animals. In particular, there is potential for multidrug-resistant bacteria that can be transmitted from animals to mankind. In this study, we investigated the genotypes of E. coli strains isolated from humans and chickens using multi-locus sequence typing (MLST) and antimicrobial resistance patterns by disk diffusion method along with integron study involving antimicrobial resistance mechanisms. From July 2013 to July 2014, E. coli strains isolated from clinical specimens (n=44) and poultry chickens (n=34). ST131 (n=20) was most common in human-derived E. coli. ST752 (n=7) was most common in chicken-derived E. coli, with four isolates each for ST117, ST189, and ST69. Of the 44 E. coli strains isolated from humans, 25 of had a class 1 integron, as opposed to only 11 of 34 strains in the E. coli isolated from chickens. There were differences in genotypes and antimicrobial resistance patterns between the chicken-derived and the human-derived E. coli.

Heat Shock Protein Augmentation of Angelica gigas Nakai Root Hot Water Extract on Adipogenic Differentiation in Murine 3T3-L1 Preadipocytes

  • Lumbera, Wenchie Marie L.;Cruz, Joseph dela;Yang, Seung-Hak;Hwang, Seong Gu
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.419-427
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    • 2016
  • There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at $42^{\circ}C$ for one hour and then allowed to recover at normal incubation temperature of $37^{\circ}C$ for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to $400{\mu}g/mL$) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.

Characterizing Milk Production Related Genes in Holstein Using RNA-seq

  • Seo, Minseok;Lee, Hyun-Jeong;Kim, Kwondo;Caetano-Anolles, Kelsey;Jeong, Jin Young;Park, Sungkwon;Oh, Young Kyun;Cho, Seoae;Kim, Heebal
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.343-351
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    • 2016
  • Although the chemical, physical, and nutritional properties of bovine milk have been extensively studied, only a few studies have attempted to characterize milk-synthesizing genes using RNA-seq data. RNA-seq data was collected from 21 Holstein samples, along with group information about milk production ability; milk yield; and protein, fat, and solid contents. Meta-analysis was employed in order to generally characterize genes related to milk production. In addition, we attempted to investigate the relationship between milk related traits, parity, and lactation period. We observed that milk fat is highly correlated with lactation period; this result indicates that this effect should be considered in the model in order to accurately detect milk production related genes. By employing our developed model, 271 genes were significantly (false discovery rate [FDR] adjusted p-value<0.1) detected as milk production related differentially expressed genes. Of these genes, five (albumin, nitric oxide synthase 3, RNA-binding region (RNP1, RRM) containing 3, secreted and transmembrane 1, and serine palmitoyltransferase, small subunit B) were technically validated using quantitative real-time polymerase chain reaction (qRT-PCR) in order to check the accuracy of RNA-seq analysis. Finally, 83 gene ontology biological processes including several blood vessel and mammary gland development related terms, were significantly detected using DAVID gene-set enrichment analysis. From these results, we observed that detected milk production related genes are highly enriched in the circulation system process and mammary gland related biological functions. In addition, we observed that detected genes including caveolin 1, mammary serum amyloid A3.2, lingual antimicrobial peptide, cathelicidin 4 (CATHL4), cathelicidin 6 (CATHL6) have been reported in other species as milk production related gene. For this reason, we concluded that our detected 271 genes would be strong candidates for determining milk production.

The effect of progressive tensional force on mRNA expression of osteoprotegerin and receptor activator of nuclear factor ${\kappa}B$ ligand in the human periodontal ligament cell (기계적 자극이 치주인대 세포의 osteoprotegerin과 receptor activator of nuclear factor ${\kappa}B$ ligand mRNA 발현에 미치는 영향)

  • Lee, Kie-Joo;Lee, Syng-Ill;Hwang, Chung-Ju;Ohk, Seung-Ho;Tian, Yu-Shin
    • The korean journal of orthodontics
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    • v.35 no.4 s.111
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    • pp.262-274
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    • 2005
  • Tooth movement is a result of mutual physiologic responses between the periodontal ligament and alveolar bone stimulated by mechanical strain. The PDL cell and osteoblast are known to have an influence on bone formation by controlling collagen synthesis and alkaline phosphatase activation. Moreover. recent studies have shown that the PDL cell and osteoblast release osteoprotegerin (OPG) and the receptor activator of nuclear factor ぉ ligand (RANKL) to control the level of osteoclast differentiation and activation which in turn influences bone resorption. In this study. progressively increased, continuous tensional force was applied to PDL cells. The objective was to find out which kind of biochemical reactions occur after tensional force application and to illuminate the alveolar bone resorption and apposition mechanism. Continuous and progressively increased tensile force was applied to PDL cells cultured on a petriperm dish with a flexible membrane The amount of $PGE_2$ and ALP synthesis were measured after 1, 3, 0 and 12 hours of force application. Secondly RT-PCR analysis was carried out for OPG and RANKL which control osteoclast differentiation and MMP-1 -8, -9, -13 aud TIMP-1 which regulate the resolution of collagen and resorption of the osteoid layer According to the results. we concluded that progressively increased, concluded force application to human PDL cells reduces $PGE_2$ synthesis, and increases OPG mRNA expression.

Mechanisms of Korean red ginseng and herb extracts(KTNG0345) for anti-wrinkle activity (홍삼 생약 복합물(KTNG0345)의 피부 주름개선에 관한 작용기전)

  • So, Seung-Ho;Lee, Seong-Kye;Hwang, Eui-Il;Koo, Bon-Suk;Han, Gyeong-Ho;Chung, Jin-Ho;Lee, Min-Jung;Kim, Na-Mi
    • Journal of Ginseng Research
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    • v.32 no.1
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    • pp.39-47
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    • 2008
  • UV irradiation causes skin-aging involving coarse wrinkles, thickening, dyspigmentation, and rough skin surface. These phenomena in complex skin tissue is controlled with receptor of cell surface growth factor and cytokine receptors. The activation of receptors induces multiple downstream signaling pathways including expression of MMPs (matrix metalloproteinases). This study was aimed to elucidate the mechanism for anti-wrinkle activity of Korean red ginseng, Torilis fructus and Corni fructus mixture (KTNG0345). In this animal study, we have investigated decreasing effects of Korean red ginseng mixture on MMP-3 synthesis through diminishing $TNF-{\alpha}$ signaling that express MMP-1, -3, and -9. c-Jun and c-fos as a component of transcription factor AP-1 (activator protein-1) were analyzed the expression level using real time PCR and western blotting. c-Jun was decreased dose dependent manner both gene and protein level where as cfos was not changed. In upstream, JNK and PAK was not changed, but p38 was decreased in down stream. MMP-3, final product in this pathway was significantly decreased in dose dependent manner. These results suggest that Korean red ginseng mixture have a anti-wrinkle activity through $TNF-{\alpha}$ mediated MMPs expression pathway.

Are Spinal GABAergic Elements Related to the Manifestation of Neuropathic Pain in Rat?

  • Lee, Jae-Hee;Back, Seung-Keun;Lim, Eun-Jeong;Cho, Gyu-Chong;Kim, Myung-Ah;Kim, Hee-Jin;Lee, Min-Hee;Na, Heung-Sik
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.2
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    • pp.59-69
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    • 2010
  • Impairment in spinal inhibition caused by quantitative alteration of GABAergic elements following peripheral nerve injury has been postulated to mediate neuropathic pain. In the present study, we tested whether neuropathic pain could be induced or reversed by pharmacologically modulating spinal GABAergic activity, and whether quantitative alteration of spinal GABAergic elements after peripheral nerve injury was related to the impairment of GABAergic inhibition or neuropathic pain. To these aims, we first analyzed the pain behaviors following the spinal administration of GABA antagonists ($1{\mu}g$ bicuculline/rat and $5{\mu}g$ phaclofen/rat), agonists ($1{\mu}g$ muscimol/rat and $0.5{\mu}g$ baclofen/rat) or GABA transporter (GAT) inhibitors ($20{\mu}g$ NNC-711/rat and $1{\mu}g$ SNAP-5114/rat) into naive or neuropathic animals. Then, using Western blotting, PCR or immunohistochemistry, we compared the quantities of spinal GABA, its synthesizing enzymes (GAD65, 67) and its receptors (GABAA and GABAB) and transporters (GAT-1, and -3) between two groups of rats with different severity of neuropathic pain following partial injury of tail-innervating nerves; the allodynic and non-allodynic groups. Intrathecal administration of GABA antagonists markedly lowered tail-withdrawal threshold in naive animals, and GABA agonists or GAT inhibitors significantly attenuated neuropathic pain in nerve-injured animals. However, any quantitative changes in spinal GABAergic elements were not observed in both the allodynic and non-allodynic groups. These results suggest that although the impairment in spinal GABAergic inhibition may play a role in mediation of neuropathic pain, it is not accomplished by the quantitative change in spinal elements for GABAergic inhibition and therefore these elements are not related to the generation of neuropathic pain following peripheral nerve injury.

Association Study of a Norepinephrine Transporter T-182C Polymorphism and Anxiety-Related Traits (불안관련특성과 노르에피네프린 수송체 T-182C 유전자 다형성의 연관연구)

  • Lim, Se-Won;Woo, Hee-Yeon;Kim, Kye-Hyun
    • Korean Journal of Psychosomatic Medicine
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    • v.16 no.1
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    • pp.47-51
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    • 2008
  • Objectives : The aim of this study was to investigate the relationship between the norepinephrine(NET) transporter T-182C polymorphism and anxiety-related traits in Korean adolescent females. Methods : One hundred sixty-nine Korean adolescent females were tested for the NET T-182C polymorphism by PCR based methods; anxiety-related traits were evaluated using the anxiety sensitivity index(ASI) and the trait form of the Spielberg State-Trait Anxiety inventory(STAI-T). Results : Scores of anxiety related traits were not different between genotypes. Comparison between T allele carries and non carriers revealed no significant difference. Conclusion : These findings suggest that the NET T-182C polymorphism is not associated with anxiety-related traits in Korean female adolescents.

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Antimicrobial Effects of Retort and Gamma Irradiation on Bacterial Populations in Spicy Chicken Sauce (레토르트 및 감마선 조사에 의한 화닭 덮밥 소스의 미생물 제어 효과 비교)

  • Kim, Young-Sik;Kim, Hyun-Joo;Yoon, Yo-Han;Shin, Myung-Gon;Kim, Cheon-Jei;Shin, Mee-Hye;Lee, Ju-Woon
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.141-147
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    • 2010
  • This study evaluated the antimicrobial effects of retort process and gamma irradiation on reduction of total bacterial populations in spicy chicken sauce, which is served on top of the steamed rice. Commercial spicy chicken sauce was treated with retort and gamma ray at 0, 1, 3, 5, and 10 kGy. Total aerobic bacterial populations were then enumerated on plate count agar and isolated bacteria from the test samples were identified using PCR analysis. Moreover, gamma ray sensitivity of identified bacteria was evaluated by $D_{10}$ values, and genotoxicity of gamma-irradiated samples was examined. Gamma irradiation at 3 kGy reduced total aerobic bacterial cell counts in spicy chicken sauce below detection limit, but total aerobic bacterial cell counts in test samples treated with retort were 2.1 log CFU/g. Identified bacteria from the samples were Bacillus subtilis, B. amyloiquefaciense, and B. pumils, and the $D_{10}$ values for B. subtilis and B. cereus were 0.39 ($R^2\;=\;0.921$) and 0.28 log CFU/g ($R^2\;=\;0.904$), respectively. The SOS chromotest showed that the gamma-irradiated spicy chicken sauce did not cause mutagenicity. These results indicate that gamma irradiation of spicy chicken sauce could be useful in ensuring microbial safety.

Virulence Factor Profiles of Escherichia coli Isolated from Pork and Chicken Meats Obtained from Retail Markets (소매시장에서 판매하는 돼지고기와 닭고기에서 분리된 Escherichia coli의 병원성 인자)

  • Choi, Sun-Keum;Lee, Min-Hwa;Lee, Bog-Hieu;Jung, Ji-Youn;Choi, Chang-Sun
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.148-153
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    • 2010
  • The aim of this study was to investigate the virulence factor profiles of Escherichia coli strains isolated from pork and chicken meats purchased from retail markets in Korea. From 943 pork and 142 chicken meats, 217 isolates of E. coli were cultured. The presence of 11 virulence factors (elt, estI, estII, astA, stx, cdt, cnf, agg, inve, eae, afa) were detected by polymerase chain reaction. Forty one (18.9%) of 217 E. coli isolates carried at least one virulence factor. Among 175 E. coli isolates from pork, the detection rate of astA, elt, eae, estII, estI, afa, and cnf were 6.9%, 4.6%, 4.6%, 4.0%, 2.3%, 1.1%, and 0.6%, respectively. However, stx, agg, and cdt were not detected in our isolates. Therefore, we conclude that astA is the most prevalent virulence factor in E. coli isolates contaminated in pork and chicken meats in Korea.