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Molecular Typing of Staphylococcus aureus Strains from Domestic Animals and Humans by REP-PCR Analysis (REP-PCR을 이용한 국내 사람과 동물유래 Staphylococcus aureus 분리주의 Molecular Typing)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.60-66
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    • 2005
  • To select the rapid and efficient molecular subtyping method for epidemiologic monitoring of Staphylococcus aureus (S. aureus) strains at clinical laboratory levels, a total 116 of S. aureus and MRSA (methicillin-resistant S. aureus) strains from diverse animal species [Korean cattle, goat, pig, dog, chicken, mouse] and also humans were analyzed. To evaluate the discriminatory ability (DA) of individual PCR methods, random amplified polymorphic of DNA [RAPD; 4M & RA primer], repetitive extragenic palindromic sequences PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) methods were conducted and then compared on their Simpson's index of diversity (SID) values based on the dendrogram patterns, which was produced by software program (BiolD2+ & GelCompar II). In first, RAPD using the 4M primer (SID 0.915) was expressed more higher SID value than that of RA primer (SID 0.874). 4M primer was expressed more powerful DA than RA. Both REP-PCR (SID 0.930) and ERIC-PCR (SID 0.929) methods showed much more higher DA than that of RAPD. According to the present results, both REP-PCR and ERIC-PCR among the tested analysis methods were found as the most reliable and discriminative molecular subtyping method, because they expressed the highest DA for the present S. aureus and MRSA strains.

Structural Characteristics of Expression Module of Unidentified Genes from Metagenome (메타게놈 유래 미규명 유전자의 발현에 관련된 특성분석)

  • Park, Seung-Hye;Jeong, Young-Su;Kim, Won-Ho;Kim, Geun-Joong;Hur, Byung-Ki
    • KSBB Journal
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    • v.21 no.2
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    • pp.144-150
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    • 2006
  • The exploitation of metagenome, the access to the natural extant of enormous potential resources, is the way for elucidating the functions of organism in environmental communities, for genomic analyses of uncultured microorganism, and also for the recovery of entirely novel natural products from microbial communities. The major breakthrough in metagenomics is opened by the construction of libraries with total DNAs directly isolated from environmental samples and screening of these libraries by activity and sequence-based approaches. Screening with activity-based approach is presumed as a plausible route for finding new catabolic genes under designed conditions without any prior sequence information. The main limitation of these approaches, however, is the very low positive hits in a single round of screening because transcription, translation and appropriate folding are not always possible in E. coli, a typical surrogate host. Thus, to obtain information about these obstacles, we studied the genetic organization of individual URF's(unidentified open reading frame from metagenome sequenced and deposited in GenBank), especially on the expression factors such as codon usage, promoter region and ribosome binding site(rbs), based on DNA sequence analyses using bioinformatics tools. And then we also investigated the above-mentioned properties for 4100 ORFs(Open Reading Frames) of E. coli K-12 generally used as a host cell for the screening of noble genes from metagenome. Finally, we analyzed the differences between the properties of URFs of metagenome and ORFs of E. coli. Information derived from these comparative metagenomic analyses can provide some specific features or environmental blueprint available to screen a novel biocatalyst efficiently.

Effects of Korean Red Ginseng extract on tissue plasminogen activator and plasminogen activator inhibitor-1 expression in cultured rat primary astrocytes

  • Ko, Hyun Myung;Joo, So Hyun;Kim, Pitna;Park, Jin Hee;Kim, Hee Jin;Bahn, Geon Ho;Kim, Hahn Young;Lee, Jongmin;Han, Seol-Heui;Shin, Chan Young;Park, Seung Hwa
    • Journal of Ginseng Research
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    • v.37 no.4
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    • pp.401-412
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    • 2013
  • Korean Red Ginseng (KRG) is an oriental herbal preparation obtained from Panax ginseng Meyer (Araliaceae). To expand our understanding of the action of KRG on central nervous system (CNS) function, we examined the effects of KRG on tissue plasminogen activator (tPA)/plasminogen activator inhibitor-1 (PAI-1) expression in rat primary astrocytes. KRG extract was treated in cultured rat primary astrocytes and neuron in a concentration range of 0.1 to 1.0 mg/mL and the expression of functional tPA/PAI-1 was examined by casein zymography, Western blot and reverse transcription-polymerase chain reaction. KRG extracts increased PAI-1 expression in rat primary astrocytes in a concentration dependent manner (0.1 to 1.0 mg/mL) without affecting the expression of tPA itself. Treatment of 1.0 mg/mL KRG increased PAI-1 protein expression in rat primary astrocytes to $319.3{\pm}65.9%$ as compared with control. The increased PAI-1 expression mediated the overall decrease in tPA activity in rat primary astrocytes. Due to the lack of PAI-1 expression in neuron, KRG did not affect tPA activity in neuron. KRG treatment induced a concentration dependent activation of PI3K, p38, ERK1/2, and JNK in rat primary astrocytes and treatment of PI3K or MAPK inhibitors such as LY294002, U0126, SB203580, and SP600125 (10 ${\mu}M$ each), significantly inhibited 1.0 mg/mL KRG-induced expression of PAI-1 and down-regulation of tPA activity in rat primary astrocytes. Furthermore, compound K but not other ginsenosides such as Rb1 and Rg1 induced PAI-1 expression. KRG-induced up-regulation of PAI-1 in astrocytes may play important role in the regulation of overall tPA activity in brain, which might underlie some of the beneficial effects of KRG on CNS such as neuroprotection in ischemia and brain damaging condition as well as prevention or recovery from addiction.

Effects of Cultured Wild Ginseng Roots on the Alcoholic Fermentation (산삼배양근 첨가가 알콜 발효에 미치는 영향)

  • Jeong Heon-Sang;Kang Tae-Su;Woo Koan-Sik;Paek Kee-Yeoup;Yu Kee-Won;Yang Seung-Joon
    • Food Science and Preservation
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    • v.12 no.4
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    • pp.402-410
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    • 2005
  • In order to manufacture the alcoholic drinks using cultured wild ginseng roots(CWGR) of 5 and $10\%$ (w/v), sugar content of fermentation media was adjusted to 24-25 $^{\circ}$Brix with white sugar and glucose. And 3 kinds of yeast (S. cerevisiae(KCCM 50757), S. cerevisiae (KCCM 50583) and S. bayanus(ATCC 10601) were used and then the quality of alcoholic drinks was analyzed by physical, chemical and sensory evaluation. Alcohol content was highest value of $15.8\%$ in $10\%$ of CWGR, white sugar, and S. bayanus(ATCC 10601). Major alcohols were ethanol and 1-propanol. Number of yeast cells increased to 5 days fermentation and slightly decreased afterwards. The pH was decreased abruptly from 5.0 in initial fermentation to 3.1-4.1 in 5 days fermentation. Total sugar contents were decreased continuously with fermentation periods and showed 7.0-10.5 $^{\circ}$Brix in 20 days fermentation. Saponin patterns and contents were various and higher in wine treated with S. bayanus(ATCC 10601). From the sensory evaluation, the highest score of overall quality was observed in the alcoholic beverage of $10\%$(w/v) of CWGR, glucose, and S. cerevisiae(KCCM 50583).

Perioperative stress prolong post-surgical pain via miR-339-5p targeting oprm1 in the amygdala

  • Zhu, Yi;Sun, Mei;Liu, Peng;Shao, Weidong;Xiong, Ming;Xu, Bo
    • The Korean Journal of Pain
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    • v.35 no.4
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    • pp.423-432
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    • 2022
  • Background: The decreased expression of mu-opioid receptors (MOR) in the amygdala may be a key molecular in chronic post-surgical pain (CPSP). It is known that miR-339-5p expression in the amygdala of a stressed rat model was increased. Analyzed by RNAhybrid, miR-339-5p could target opioid receptor mu 1 (oprm1) which codes MOR directly. So, the authors hypothesized that miR-339-5p could regulate the expression of MOR via targeting oprm1 and cause the effects to CPSP. Methods: To simulate perioperative short-term stress, a perioperative stress prolongs incision-induced pain hypersensitivity without changing basal pain perception rat model was built. A pmiR-RB-REPORTTM dual luciferase assay was taken to verify whether miR-339-5p could act on oprm1 as a target. The serum glucocorticoid level of rats was test. Differential expressions of MOR, GFAP, and pERK1/2 in each group of the rats' amygdala were tested, and the expressions of miR-339-5p in each group of rats' amygdalas were also measured. Results: Perioperative stress prolonged the recovery time of incision pain. The expression of MOR was down-regulated in the amygdala of rats in stress + incision (S + IN) group significantly compared with other groups (P < 0.050). miR-339-5p was up-regulated in the amygdala of rats in group S + IN significantly compared with other groups (P < 0.050). miR-339-5p acts on oprm1 3'UTR and take MOR mRNA as a target. Conclusions: Perioperative stress could increase the expression of miR-339-5p, and miR-339-5p could cause the expression of MOR to decrease via targeting oprm1. This regulatory pathway maybe an important molecular mechanism of CPSP.

Petrochemistry and Environmental Geochemistry of Shale and Coal from the Daedong Supergroup, Chungnam Coal Field, Korea (충남탄전, 대동누층군의 셰일과 탄질암에 관한 암석화학 및 환경지구화학적 특성)

  • Lee, Chan Hee;Lee, Hyun Koo;Kim, Kyoung-Woong
    • Economic and Environmental Geology
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    • v.30 no.5
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    • pp.417-431
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    • 1997
  • Characteristics of sedimentary rocks and enrichment of toxic elements in shale and coal from the Chungnam coal field were investigated based upon geochemistry of major, trace and rare earth elements. Shale and coal of the area are interbedded along the Traissic to the Jurassic Daedong Supergroup, which can be subdivided into grey shale, black shale and coal. The coal had been mined, however all the mines are abandonded due to the economic problems. The shale and coal are characterized by relatively low contents of $SiO_2$, and $Al_2O_3$ and high levels of loss-on-ignition (LOI), CaO and $Na_2O$ in comparison with the North American Shale Composite (NASC). Light rare earth elements (La, Ce, Yb and Lu) are highly enriched with the coal. Ratios of $Al_2O_3/Na_2O$ and $K_2O/Na_2O$ in shale and coal range from 30.0 to 351.8 and from 4.2 to 106.8, which have partly negative correlations against $SiO_2/Al_2O_3$ (1.24 to 6.06), respectively. Those are suggested that controls of mineral compositions in shale and coal can be due to substitution and migration of those elements by diagenesis and metamorphism. Shale and coal of the area may be deposited in terrestrial basin deduced from high C/S (39 to 895) and variable composition of organic carbon (0.39 to 18.40 wt.%) and low contents of reduced sulfur (0.01 to 0.05 wt.%). These shale and coal were originated from the high grade metamorphic and/or igneous rocks, and the rare earth elements of those rocks are slightly influenced with diagenesis and metamorphism on the basis of $Al_2O_3$ versus La, La against Ce, Zr versus Yb, the ratios of La/Ce (0.38 to 0.85) and Th/U (3.6 to 14.6). Characteristics of trace and rare earth elements as Co/Th (0.07 to 0.86), La/Sc (0.31 to 11.05), Se/Th (0.28 to 1.06), V/Ni (1.14 to 3.97), Cr/V (1.4 to 28.3), Ni/Co (2.12 to 8.00) and Zr/Hf (22.6~45.1) in the shale and coal argue for inefficient mixing of the simple source lithologies during sedimentation. These rocks also show much variation in $La_N/Yb_N$ (1.36 to 21.68), Th/Yb (3.5 to 20.0) and La/Th (0.31 to 7.89), and their origin is explained by derivation from a mixture of mainly acidic igneous and metamorphic rocks. Average concentrations in the shale and coal are As=7.2 and 7.5, Ba=913 and 974, Cr=500 and 145, Cu=20 and 26, Ni=38 and 35, Pb=30 and 36, and Zn=77 and 92 ppm, respectively, which are similar to those in the NASC. Average enrichment indices for major elements in the shale (0.79) and coal (0.77) are lower than those in the NASC. In addition, average enrichment index for rare earth elements in coal (2.39) is enriched rather than the shale (1.55). On the basis of the NASC, concentrations of minor and/or environmental toxic elements in the shale and coal were depleted of all the elements examined, excepting Cr, Pb, Rb and Th. Average enrichment indices of trace and/or potentially toxic elements (As, Cr, Cu, Ni, Pb, U and Zn) are 1.23 to 1.24 for shale and 1.06 to 1.22 for coal, respectively.

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Effects of Ginsenosides and Their Metabolites on Voltage-dependent Ca2+ Channel Subtypes

  • Lee, Jun-Ho;Jeong, Sang Min;Kim, Jong-Hoon;Lee, Byung-Hwan;Yoon, In-Soo;Lee, Joon-Hee;Choi, Sun-Hye;Lee, Sang-Mok;Park, Yong-Sun;Lee, Jung-Ha;Kim, Sung Soo;Kim, Hyoung-Chun;Lee, Boo-Yong;Nah, Seung-Yeol
    • Molecules and Cells
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    • v.21 no.1
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    • pp.52-62
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    • 2006
  • In previous reports we demonstrated that ginsenosides, active ingredients of Panax ginseng, affect some subsets of voltage-dependent $Ca^{2+}$ channels in neuronal cells expressed in Xenopus laevis oocytes. However, the major component(s) of ginseng that affect cloned $Ca^{2+}$ channel subtypes such as ${\alpha}_{1C}$(L)-, ${\alpha}_{1B}$(N)-, ${\alpha}_{1A}$(P/Q)-, ${\alpha}_{1E}$(R)- and ${\alpha}_{1G}$(T) have not been identified. Here, we used the two-microelectrode voltage clamp technique to characterize the effects of ginsenosides and ginsenoside metabolites on $Ba^{2+}$ currents ($I_{Ba}$) in Xenopus oocytes expressing five different $Ca^{2+}$ channel subtypes. Exposure to ginseng total saponins (GTS) induced voltage-dependent, dose-dependent and reversible inhibition of the five channel subtypes, with particularly strong inhibition of the ${\alpha}_{1G}$-type. Of the various ginsenosides, $Rb_1$, Rc, Re, Rf, $Rg_1$, $Rg_3$, and $Rh_2$, ginsenoside $Rg_3$ also inhibited all five channel subtypes and ginsenoside $Rh_2$ had most effect on the ${\alpha}_{1C}$- and ${\alpha}_{1E}$-type $Ca^{2+}$ channels. Compound K (CK), a protopanaxadiol ginsenoside metabolite, strongly inhibited only the ${\alpha}_{1G}$-type of $Ca^{2+}$ channel, whereas M4, a protopanaxatriol ginsenoside metabolite, had almost no effect on any of the channels. $Rg_3$, $Rh_2$, and CK shifted the steady-state activation curves but not the inactivation curves in the depolarizing direction in the ${\alpha}_{1B}$- and ${\alpha}_{1A}$-types. These results reveal that $Rg_3$, $Rh_2$ and CK are the major inhibitors of $Ca^{2+}$ channels in Panax ginseng, and that they show some $Ca^{2+}$ channel selectivity.

A Statistical Analysis of the Fereign Bodies in the Food and air Passages (식도 및 기도이물의 임상통계학적 고찰)

  • 정해영;권평중;박성준;민양기;김중환
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1978.06a
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    • pp.4.1-4
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    • 1978
  • The foreign bodies in the food and air passages are frequently observed in the field of otolaryngology, and the foreign bodies in the air passages have much significance in clinical practice because they may cause sudden death. A statistical study was done on 95 cases of foreign bodies in the food and air passages who had visited department of otolaryngology, Chung-Ang university from June, 1968 to April, 1978. The results were as follows; 1. The total cases of foreign bodies in the food and air passages was 95; 89 cases (93.7%) were in the food passage and the remaining (6.3%) were in the air passage. The ratio between the food passage and air passage was about 14. 8 to 1.0. 2. In distribution by sex, 64 cases (67.4%) were in male patients and the remaining 31 cases (32.6%) were in female patients. The ratio between male and female was 2.1 to 1.0. 3. The kinds of foreign bodies in the food in the order of their frequency, were coin, gogame stone and pebble. In the air passages, the peanut and bean were most frequently found. 4. In distribution by age, 64 cases (67.4%) of all foreign bodies were found in children under 5 year old, and coin was the most common kind of foreign body. Except for meat, almost all of foreign bodies were found in children under 10 year old. 5. There was chronologically no significant tendency in incidence; the incidence, however, had decreased during recent 2 years. 6. In the location of foreign bodies in the food passage, 70 cases (78.7%) were found at the first esophageal narrowing. Meat was more frequently found at second esophageal narrowing associated with cicatrical stenosis. Almost all of foreign bodies in the air passage was found in bronchi; 3 cases were in the right side of bronchi, and 2 cases were in the left side of bronchi and 1 case was in glottic region. 7. In duration of lodgement, 50 cases (52.6%) visited our hospital within 24 hours, and 3 cases after 30 days. 8. Under topical anesthesia, 83 cases (93.3%) of the foreign bodies in the food passage were removed by esophagoscopy, by 6 cases (6.7%) failed to be removed. All of the foreign bodies of the air passages (6 cases) were removed by bronchoscopy. Among them, 5 cases under 5 years old were removed by inferior bronchoscopy through tracheostomy site.

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Transcriptome Analyses for the Anti-Adipogenic Mechanism of an Herbal Composition (생약복합물의 지방세포형성억제 기전규명을 위한 전사체 분석)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Bae, Sung-Min;Chae, Soo-Ahn;Lee, Jung-Ju;Oh, Dong-Jin;Park, Suk-Won;Cho, Soo-Hyun;Shim, Yae-Jie;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1054-1065
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    • 2010
  • SH21B is a natural composition composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). In our previous study, we reported that SH21B inhibited adipogenesis and fat accumulation in 3T3-L1 cells through modulation of various regulators in the adipogenesis pathway. The aim of this study was to analyze the transcriptome profiles for the anti-adipogenic effects of SH21B in 3T3-L1 cells. Total RNAs from SH21B-treated 3T3-L1 cells were reverse-transcribed into cDNAs and hybridized to Affymetrix Mouse Gene 1.0 ST array. From microarray analyses, we identified 2,568 genes of which expressions were changed more than two-fold by SH21B, and the clustering analyses of these genes resulted in 9 clusters. Three clusters among the 9 showed down-regulation by SH21B (cluster 4, cluster 6 and cluster 9), and two clusters showed up-regulation by SH21B (cluster 7 and cluster 8) during the adipogenesis of 3T3-L1 cells. It was found that many genes related to cell proliferation and adipogenesis were included in these clusters. Clusters 4, 6 and 9 included genes which were related with adipogenesis induction and cell cycle arrest. Clusters 7 and 8 included genes related to cell proliferation as well as adipogenesis inhibition. These results suggest that the mechanisms of the anti-adipogenic effects of SH21B may be the modulation of genes involved in cell proliferation and adipogenesis.

Effect of Microbial Phytase in Low Phosphorus and Calcium Level Diet on the Performance and Nutrient Digestibility in Laying Hens (인과 칼슘의 수준이 낮은 산란계 사료 내 미생물 Phytase의 첨가가 생산성 및 영양소 소화율에 미치는 영향)

  • Min B.J.;Kwon O.S.;Lee W.B.;Son K.S.;Hong J.W.;Yang S.J.;Moon T.H.;Kim I.H.
    • Korean Journal of Poultry Science
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    • v.32 no.1
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    • pp.15-21
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    • 2005
  • This Study was conducted to investigate the effects of microbial phytase in low phosphirus and calcium level diet on the performance and nutrient digestibility in laying hens. One hundred ninety two, 50 wks old, ISA brown commerical layers were used for 12 weeks feeding trial after 7-d adjustment period. Four dietary treatments included CON(control; Co.), P2 ($0.06\%$ Natuphos, BASF) and P3 ($0.06\%$ PHOSMAX, GENOFOCUS). Ca and available P concentrations of P1, P2 and P3 were 90 and $50\%$ of NRC recommecdations to accentuate difference in response to phytase availability. In whole period, egg production was not affected by treatments. At 12 weeks, egg weight was significantly increased in adding phytase treatments (P<0.05). Egg shell thickness was increased in P1, P2 and P3 treatments compared with control (P<0.05) at 9 weeks. Ca concentration of serum tended to decrease in P1 treatment without significant difference (P>0.05). Ca and P concentrations of tibia were higher in layers fed dietary phyrase than those fed control diet without significant difference (P>0.05). Digestibilities of DM, N and ash were improved in P1 treatment compared with P2 and P3 treatments (P<0.05). Ca and P digestibilities were the highest in P2 treatment (P>0.05), but was not significant difference between control and P1 treatments.