• Title/Summary/Keyword: IS-PCR

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Trends in Rapid Detection Methods for Food-borne pathogenic Microorganisms by Using New Technologies (신기술 이용 식중독균 신속검출법 개발 동향 분석)

  • Kim, Hyun-Joo;Kim, Yong-Soo;Chung, Myung-Sub;Oh, Deog-Hwan;Chun, Hyang-Sook;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.376-387
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    • 2010
  • Recently, speedy, convenient and easy detection technologies have been developed rapidly and on the contrary, studies on development of traditional detectors applying biochemical characteristics has gradually been decreased. This review examined trend in current studies on detection of food-borne pathogenic microorganisms in the fields of selective media, immuno-assay, Polymerase Chain Reaction (PCR), microarray, terahertz spectroscopy & imagination and so on. Most traditional methods to detect the organisms from food matrix rely on selective media and such a method have disadvantages like long time requirement and distinguishing one species only from each selective medium although they are highly economical. Various new convenient methods such as Enzyme Linked Immuno-sorbent Assay (ELISA), paper-strip kit, fluoroimmunoassay etc. have been developed. The most ideal method for detecting food-borne pathogenic microorganisms in foods should be accurate, convenient, rapid and economical. Additionally, it is needed that capabilities of quantitative analysis and automation to be applied to industries.

Effects of Moxi-tar Herbal Acupuncture of LI11 on inflammatory bowel disease induced by TNBS in mice (TNBS로 유도된 대장염에서 곡지의 구진약침 효과)

  • Song, Moon-Young;Park, Sang-Yeon;Kim, Jae-Hyo;Ahn, Sung-Hun;Kim, Kyoung-Sik;Sohn, In-Chul
    • Korean Journal of Acupuncture
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    • v.25 no.3
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    • pp.147-166
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    • 2008
  • Objectives : Crohn's disease (CD) is characterized by a chronic relapsing inflammation of the bowel in which proinflammatory cytokines play an important perpetuating role. Methods : Mice (preventive animal model of gliotoxin) were treated with 5 % 2,4,6-trinitrobenzenesulfonic acid (TNBS) at day 1 and day 7. To investigate preventive effects of acupuncture with Gujin at $LI_{11}$, acupuncture was carried out at day -1, day 1, day 3. And, to investigate therapeutic effects, acupuncture with Gujin was carried out at day 3, day 5, day 7. For the data analysis, we checked weight and width of colon, diarrhea, edema, survival rate, changes of body weight, and myeloperoxygenase (MPO) activity. For analysing protein expression, we carried out immunohistochemical staining and Western blot and we analyzed mRNA expression by RT-PCR. Results : Colon of TNBS treated mice was erosive and shortening compared with the colon of control mice and induced damages of colon epithelial cell layer and induced infiltration of immune cells in all layer of colon. Acupuncture of gujin at $LI_{11}$ in preventive mode suppressed macorscopic damages such as erosive and shortening of colon by TNBS and damages of intestinal epithelial cells and infiltration of immune cells in the colon. The average weight of 5 cm distal colon was increased in TNBS treated mice (758${\mu}g$) compared with in control mice (112${\mu}g$) and width of distal colon was also increased in TNBS treated mice (4.9mm) compared with in control mice (1.3mm). Acupuncture with Gujin at $LI_{11}$ in preventive and therapeutic mode suppressed increase of colon weight and width by TNBS. TNBS induced edema of colon and diarrhea and Acupunctured with Gujin at $LI_{11}$ in preventive and therapeutic mode ameliorated these symptom by TNBS. In preventive and therapeutic mode, the effects of acupuncture with Gujin at $LI_{11}$ were increasing the motility, suppressing body weight decreasing, suppressing MPO activity, reducing expressing of TNF-${\alpha}$, IL-1b, and ICAM-1 in colon compared with that by TNBS Conclusions : This study demonstrates that acupuncture with Gujin at $LI_{11}$ represents a potential therapeutic method of Crohn's disease.

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Studies on Proteolytic and Fibrinolytic Activity of Bacillus subtilis JM-3 Isolated from Anchovy Sauce (멸치액젓으로부터 분리한 Bacillus subtilis JM-3의 단백질 분해활성과 혈전 용해 활성에 관한 연구)

  • Lee, Sang-Soo;Kim, Sang-Moo;Park, Uk-Yeon;Kim, Hee-Yun;Shin, Il-Shik
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.283-289
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    • 2002
  • This study was performed to search for potential microorganism that has rapid fermenting and physiological function from anchovy sauce. We isolated three bacterial strains, JM-1, JM-2, and JM-3 with proteolytic and fibrinolytic activity from anchovy sauce. Among the 3 bacterial strains, JM-3 showed the strongest proteolytic and fibrinolytic activity. Bacterial strain JM-3 was gram-positive rod, motile and formed endospore. The 16S rRNA of bacterial strain JM-3 was amplified by PCR and then its sequence was determined by ABI 310 genetic analyzer. The 16S rRNA sequence of bacterial strain JM-3 was compared to BLAST DNA database and identified to Bacillus subtilis with 99% of homology. The optimum temperature, pH and NaCl concentration for growth of B. subtilis JM-3 were $40^{\circ}C$, 5.0 and 0%, respectively. The optimum temperature, pH and NaCl concentration for proteolytic and fibrinolytic enzyme production of B. subtilis JM-3 were same as optimum conditions for growth. At 20% of NaCl concentration which is common NaCl concentration of fish sauce, B. subtilis JM-3 showed about 60% of proteolytic and fibrinolytic activity of 0% NaCl concentration. From above results, we found that B. subtilis JM-3 will be able to used for starter of functional fish sauce.

Polymorphisms of Angiotensin-converting Enzyme Gene Associated in Patients with COPD with or without Bronchial Hyperresponsiveness (만성폐쇄성폐질환에서 ACE 유전자 다형성과 기관지 과민성의 연관성)

  • Kim, Seung Soo;Choi, Eu Gene;Park, Seoung Ju;Lee, Heung Bum;Lee, Yong Chul;Rhee, Yang Keun
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.25-30
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    • 2005
  • Background : An insertion-deletion polymorphism of angiotensin converting enzyme (ACE) gene has been shown to be associated with enzyme activity levels of ACE. Reported results that have been mutually contradictory about asthmatic hypersensitiveness and occurrence according to ACE gene insertion (I)/deletion (D) polymorphism. Also, the involvement of the ACE genes as the genetic basis of bronchial asthma is currently controversy. We investigated whether there was any association between polymorphisms of the ACE genes and airway hyper-responsiveness in chronic obstructive pulmonary disease (COPD). Methods : A total of 100 patients with COPD were enrolled in this study. The ACE genotypes were determined in all subjects by polymerase chain reaction. Pulmonary function test including bronchodilator response (BDR), methacholine bronchial provocation test (MBPT) were done in those patients. Airway hyper-responsiveness include any findings of positive BDR or MBPT. Results : In COPD patients, the ACE genotype distribution did not differ significantly among groups of patients with severities of COPD, and with or without airway hyper-responsiveness. Conclusions : These results suggest that polymorphisms of the ACE gene may not be associated with airway hyper-responsiveness, development and severity of COPD.

Mitochondrial Genome Sequence of Echinostoma revolutum from Red-Crowned Crane (Grus japonensis)

  • Ran, Rongkun;Zhao, Qi;Abuzeid, Asmaa M.I.;Huang, Yue;Liu, Yunqiu;Sun, Yongxiang;He, Long;Li, Xiu;Liu, Jumei;Li, Guoqing
    • Parasites, Hosts and Diseases
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    • v.58 no.1
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    • pp.73-79
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    • 2020
  • Echinostoma revolutum is a zoonotic food-borne intestinal trematode that can cause intestinal bleeding, enteritis, and diarrhea in human and birds. To identify a suspected E. revolutum trematode from a red-crowned crane (Grus japonensis) and to reveal the genetic characteristics of its mitochondrial (mt) genome, the internal transcribed spacer (ITS) and complete mt genome sequence of this trematode were amplified. The results identified the trematode as E. revolutum. Its entire mt genome sequence was 15,714 bp in length, including 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and one non-coding region (NCR), with 61.73% A+T base content and a significant AT preference. The length of the 22 tRNA genes ranged from 59 bp to 70 bp, and their secondary structure showed the typical cloverleaf and D-loop structure. The length of the large subunit of rRNA (rrnL) and the small subunit of rRNA (rrnS) gene was 1,011 bp and 742 bp, respectively. Phylogenetic trees showed that E. revolutum and E. miyagawai clustered together, belonging to Echinostomatidae with Hypoderaeum conoideum. This study may enrich the mitochondrial gene database of Echinostoma trematodes and provide valuable data for studying the molecular identification and phylogeny of some digenean trematodes.

Analysis of the Gene Expression by Laser Capture Microdissection (III) -Microarray Analysis of the Gene Expression at the Mouse Uterine Luminal Epithelium of the Implantation Sites during Apposition Period- (Laser Capture Microdissection을 이용한 유전자 발현 연구 (III) -생쥐 착상 부위 자궁 내강상피 조직에서 배아 병치 기간 동안 일어나는 유전자 발현에 관한 Microarray 분석-)

  • Yoon, Se-Jin;Jeon, Eun-Hyun;Park, Chang-Eun;Ko, Jung-Jae;Choi, Dong-Hee;Cha, Kwang-Yul;Kim, Se-Nyun;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.4
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    • pp.323-335
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    • 2002
  • Object: The present study was accomplished to obtain a gene expression profile of the luminal epithelium during embryo apposition in comparison of implantation (1M) and interimplantation (INTER) sites. Material and Method: The mouse uterine luminal epithelium from IM and INTER sites were sampled on day 4.5 (Day of vaginal plug = day 0.5) by Laser Captured Microdissection (LCM). RNA was extracted from LCM captured epithelium, amplified, labeled and hybridized to microarrays. Results from microarray hybridization were analyzed by Significance Analysis of Microarrays (SAM) method. Differential expression of some genes was confirmed by LCM followed by RT-PCR. Results: Comparison of IM and INTER sites by SAM identified 73 genes most highly ranked at IM, while 13 genes at the INTER sites, within the estimated false discovery rate (FDR) of 0.163. Among 73 genes at IM, 20 were EST/unknown function, and the remain 53 were categorized to the structural, cell cycle, gene/protein expression, immune reaction, invasion, metabolism, oxidative stress, and signal transduction. Of the 24 structural genes, 14 were related especially to extracellular matrix and tissue remodeling. Meanwhile, among 13 genes up-regulated at INTER, 8 genes were EST/unknown function, and the rest 5 were related to metabolism, signal transduction, and gene/protein expression. Among these 58 (53+5) genes with known functions, 13 genes (22.4%) were related with $Ca^{2+}$ for their function. Conclusions: Results of the present study suggest that 1) active tissue remodeling is occurring at the IM sites during embryo apposition, 2) the INTER sites are relatively quiescent than IM sites, and 3) the $Ca^{2+}$ may be a crucial for apposition. Search for human homologue of those genes expressed in the mouse luminal epithelium during apposition will help to understand the implantation process and/or implantation failure in humans.

Effect of PLGA/Silk Fibroin Hybrid Film on Attachment and Proliferation of Schwann Cells (실크피브로인을 함유한 PLGA 하이브리드 필름이 슈반세포의 부착과 증식에 미치는 영향)

  • Kim, Hye-Lin;Yoo, Han-Na;Park, Hyun-Jin;Kim, Yong-Gi;Lee, Dong-Won;Kang, Young-Sun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.35 no.1
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    • pp.7-12
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    • 2011
  • Poly(lactic-co-glycolic acid) (PLGA) is a biodegradable synthetic polymer with acceptable mechanical strength and well-controlled degradation rate. Also, it can be easily fabricated into many shapes. Silk fibroin contains powerful bioactive molecules. We fabricated natural/synthetic hybrid films using 0, 10, 20, 40 and 80 wt% of silk fibroin. Schwann cells (SCs) were seeded on PLGA/silk fibroin hybrid films and confirmed the effects of adhesion and proliferation on SCs according to the content of silk fibroin. In this study, we confirmed PLGA/silk fibroin hybrid film containing 40% and 80% of silk fibroin interrupted adhesion and proliferation of SCs. Films containing 10% and 20% of silk, however, provided suitable environment for growth and proliferation of SCs. These results suggest that silk fibroin provides suitables surface to neural cells and its proper content provides proper culture conditions to improve cell adhesion and proliferation.

Quality characteristics of buckwheat Soksungjang manufactured by Bacillus subtilis HJ18-4 (Bacillus subtilis HJ18-4를 이용하여 제조한 메밀 속성장의 품질특성)

  • Park, Na Young;Lee, Sun Young;Kim, Ji Yeun;Choi, Hye Sun
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.699-704
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    • 2013
  • Buckwheat Soksungjang (BS) is a bealmijang manufactured with buckwheat and soybeans. We manufactured BS using Bacillus subtilis HJ18-4 (HJ18-4), which has high enzyme activities and antibacterial effects. HJ18-4 was inoculated in a different process during the BS manufacturing, which was the meju-making time (Treat 1), and the salt water time was added (Treat 2). The physiochemical and microbial characteristics of the BS were analyzed. As a result, the total aerobic counts (7~8 log CFU/mL) in the BS increased after 15 days of fermentation. Especially, Treat 1 showed higher total aerobic counts and amino-type nitrogen (65.38~202.52 mg%) than Treat 2. During the BS fermentation, the reduction of the sugar contents and the enzyme (protease and amylase) activities decreased. In the relative quantitative expression level of PlcR, Treat 1 did not show toxin gene expressions at the end of the fermentation on Day 23. Treat 1 showed suitable B. cereus physiochemical quality characteristics and inhibition effects. When the modified-form type of fermented soybean paste was manufactured with a single starter, it could not reproduce the natural fermentation quality. These results suggest that the addition of a starter (HJ18-4) in the Meju manufacturing process could enhance the quality characteristics of the manufactured BS via natural fermentation and by suppressing B. cereus.

Korean Red Ginseng attenuates anxiety-like behavior during ethanol withdrawal in rats

  • Zhao, ZhengLin;Kim, Young Woo;Wu, YiYan;Zhang, Jie;Lee, Ju-Hee;Li, XiaoHua;Cho, Il Je;Park, Sang Mi;Jung, Dae Hwa;Yang, Chae Ha;Kim, Sang Chan;Zhao, RongJie
    • Journal of Ginseng Research
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    • v.38 no.4
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    • pp.256-263
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    • 2014
  • Background: Korean Red Ginseng (KRG) is known to have antianxiety properties. This study was conducted to investigate the anxiolytic effects of KRG extract (KRGE) during ethanol withdrawal (EW) and the involvement of the mesoamygdaloid dopamine (DA) system in it. Methods: Rats were treated with 3 g/kg/d of ethanol for 28 d, and subjected to 3 d of withdrawal. During EW, KRGE (20 mg/kg/d or 60 mg/kg/d, p.o.) was given to rats once/d for 3 d. Thirty min after the final dose of KRGE, anxiety-like behavior was evaluated in an elevated plus maze (EPM), and plasma corticosterone (CORT) levels were determined by a radioimmunoassay (RIA). In addition, concentrations of DA and 3,4-dihydroxyphenylacetic acid (DOPAC) in the central nucleus of the amygdala (CeA) were also measured by high performance liquid chromatography (HPLC). Results: The EPM test and RIA revealed KRGE inhibited anxiety-like behavior and the over secretion of plasma CORT during EW. Furthermore, the behavioral effect was blocked by a selective DA D2 receptor (D2R) antagonist (eticlopride) but not by a selective DA D1 receptor (D1R) antagonist (SCH23390). HPLC analyses showed KRGE reversed EW-induced decreases of DA and DOPAC in a dose-dependent way. Additionally, Western blotting and real-time polymerase chain reaction (PCR) assays showed that KRGE prevented the EW-induced reductions in tyrosine hydroxylase (TH) protein expression in the CeA and TH mRNA expression in the ventral tegmental area (VTA). Conclusion: These results suggest that KRGE has anxiolytic effects during EW by improving the mesoamygdaloid DA system.

Prevalence of Nosema and Virus in Honey Bee (Apis mellifera L.) Colonies on Flowering Period of Acacia in Korea

  • Hong, In-Pyo;Woo, Soon-Ok;Choi, Yong-Soo;Han, Sang-Mi;Kim, Nam-Suk;Kim, Hye-Kyung;Han, Sang-Hoon;Lee, Man-Young;Lee, Myeong-Lyeol;Byeon, Kyu-Ho
    • Mycobiology
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    • v.39 no.4
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    • pp.317-320
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    • 2011
  • Honey production from approximately 1.6 million colonies owned by about 199,000 Korean beekeepers was almost 23,000 metric tons in 2009. Nosema causes significant losses in honey production and the virus decreases population size. We initiated a survey of honey bee colonies on the blooming period of Acacia to determine the prevalence of Nosema and virus in 2011. Most Korean beekeepers have moved from the south to north of Korea to get Acacia nectar for 2 mon. This provided a valuable opportunity to sample bees originating from diverse areas in one location. Twenty hives owned by 18 beekeepers were sampled in this year. Nosema spore counts ranged from zero to 1,710,000 spores per bee. The average number of nosema spores per bee was 580,000. Approximately 95% of the colonies were infected with Nosema, based on the presence of spores in the flowering period of Acacia. This indicates that Nosema is the predominant species affecting honeybee colonies. Also, the seven most important honeybee viruses were investigated by reverse transcription-PCR. Among them, four different viruses were detected in samples. Black queen cell virus was present in all samples. Chronic bee paralysis virus was detected in 10% of samples. Deformed wing virus was present in only 5% of the samples. Prevalence of Sacbrood virus was 15%. However, Cloudy wing virus, Israel acute paralysis virus and kashmir bee virus were not detected in any of samples.