• Title/Summary/Keyword: human-to-human (H2H)

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Construction of Retrovirus Vector System for the Regulation of Recombinant hTPO Gene Expression (재조합 hTPO 유전자의 발현 조절을 위한 Retrovirus Vector System의 구축)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Do-Hyang;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.161-167
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    • 2007
  • In this study, we constructed and tested retrovirus vectors designed to express the human thrombopoietin gene under the control of the tetracycline-inducible promoters. To increase the hTPO gene expression at him-on state, WPRE sequence was also introduced into retrovirus vector at downstream region of either the hTPO gene or the sequence encoding reverse tetracycline-controlled transactivator (rtTA). Primary culture cells (PFF, porcine fetal fibroblast; CEF, chicken embryonic fibroblast) infected with the recombinant retrovirus were cultured in the medium supplemented with or without doxycycline for 48hr, and induction efficiency was measured by comparing the hTPO gene expression level using RT-PCR, western blot and ELISA. Higher hPTO expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hTPO (in CEF) or rtTA(in PFF) gene. This resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

Protective Role of Curcuma longa L. Extracts on Hydrogen Peroxide-Induced DNA Damage in Human Leukocytes (산화적 스트레스로 유도된 인체 백혈구 DNA 손상에 대한 울금 추출물의 보호효과)

  • Seo, Bo-Young;Park, Eunju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.545-551
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    • 2017
  • Curcuma longa L. (CL) is widely used as a spice and coloring agent in several foods, such as curry and mustard, as well as cosmetics and drugs. In this study, we investigated the protective effects of CL extracted with various solvents [methanol (MC), ethanol (EC), acetone (AC)] on $H_2O_2-induced$ DNA damage in human leukocytes along with total polyphenol contents (TPC) and antioxidant properties. The antioxidant effects of CL were determined by measuring 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (RSA) and superoxide dismutase (SOD)-like activity. The preventive effect of CL on oxidative stress-induced DNA damage and DNA repair capacities were assessed using comet assay. MC showed the highest TPC (11.17 g gallic acid equivalents/100 g) and antioxidant properties among the solvent extracts. The $SC_{50}$ for DPPH RSA was MC: 35.0 > AC: 45.8 > EC: $57.8{\mu}g/mL$ and SOD-like activity was MC: 46.6 > EC: 141.5 > AC: $296.4{\mu}g/mL$. In the comet assay, the $ED_{50}$ value of MC showed the highest inhibition ($86.7{\mu}g/mL$) of $H_2O_2-induced$ DNA damage, followed by AC ($110.0{\mu}g/mL$) > EC ($115.8{\mu}g/mL$). Analysis of the percentage of damaged cells showed that repair capacity significantly decreased at 4, 8, and 12 h from $H_2O_2-induced$ oxidative stress in each extract. After 12 h, level of DNA damage recovery was similar to the negative control level. These results suggest that CL has potential antioxidant activity and a protective effect against oxidation-induced DNA damage, and the methanol extract of CL was the most effective.

Antioxidant Effect of Extracts from 9 Species of Forest Plants in Korea (국내 9종 산림식물 추출물의 항산화 효능)

  • Sim, Wan-Sup;Lee, Jong Seok;Lee, Sarah;Choi, Sun-Il;Cho, Bong-Yeon;Choi, Seung-Hyun;Han, Xionggao;Jang, Gill-Woong;Kwon, Hee-Yeon;Choi, Ye-Eun;Kim, Jong-Yea;Kim, Jong-Dai;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.34 no.4
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    • pp.404-411
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    • 2019
  • This study was carried out to investigate the antioxidant effects of extracts from 9 species of forest plants in Korea. DPPH, ABTS, $NaNO_2$, hydrogen peroxide radical scavenging activity and reducing power activity were evaluated to measure the antioxidant activities of plant extracts. As a result, Geranium thunbergii has been identified as the most effective antioxidant resource. Also, total phenolic content was highest in Geranium thunbergii ($303.94{\pm}0.63mg\;GAE/g$) among 9 species extracts. Total flavonoid content was highest in Rosa multiflora ($24.32{\pm}0.22mg\;QE/g$) and proanthocyanidin content was highest in Vitis ficifolia ($279.00{\pm}4.58mg\;CE/g$) among 9 species extracts. In addition, the protective effect of plant extracts in $H_2O_2-induced$ human dermal fibroblast (HDF) cell systems were also assessed. Significant protective effects in $H_2O_2-induced$ human dermal fibroblast (HDF) cell systems were found in all plant extracts, especially in Geranium thunbergii. These results suggest that Geranium thunbergii could be a potential natural resource for antioxidant activity.

A Case Study of two Child-care Centers to Encourage Outdoor Play Environments and Play Programs in Daejeon (보육시설 실외놀이 환경과 실외놀이 프로그램 연계를 위한 기초 사례연구 - 대전시 A와 H 어린이집을 대상으로 -)

  • Choi, Mock-Wha;Son, Seung-Hee;Lim, Hyo-Sin
    • Korean Journal of Human Ecology
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    • v.19 no.5
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    • pp.775-794
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    • 2010
  • This study is to provided a basic model to encourage environment and outdoor play programs. The study was based on the analyses of actual activities performed in two child care centers in Daejeon area and the way in which the programs for outdoor play were related to the environment of the centers. Data were collected through 'The Daily Report of the Outdoor Play Programs: Plan and Evaluation' paper conducted at each center performed from 2007 to 2009. Following Mockwha Choi et al., (2007), the play observed and described by teachers was analyzed to classify play areas and play activities within each area. The places, equipment and, playthings using in each play period were also examined as well as the limitation and inspiration caused by the environment. The results of the study are as follows: 1) Empty and unfilled space tended to generate more types of non-structural play and role-pretend play 2)Adventurous play was rarely observed. More thoughtfully designed physical environments need to be provided for children to experience challenge and adventure through physical activities 3) Outdoor play environment should be suitable for the overall developments of infants and toddlers depending on their activities. A child care specialist should support the design such environment 4) Naturally, the programs of outdoor play will not be identical for all child care centers. Rather, they should go along with the programmatic characters and environmental attributes of each center. Therefore the awareness of teachers and parents as to the importance of must realize of outdoor play should be further increased.

Identification and characterization of Shiga toxin-producing Escherichia coli isolated from diarrhea in calves (송아지 설사분변으로부터 Shiga toxin-producing Escherichia coli 의 분리 및 특성규명)

  • Lim, Keum-Gi;Kang, Mun-Il;Kim, Snag-Ki;Nam, Kyung-Woo;Park, Hyun-Joo;Park, Jin- Ryang;Cho, Kyoung-Oh;Lee, Bong-Joo
    • Korean Journal of Veterinary Research
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    • v.46 no.2
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    • pp.135-142
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    • 2006
  • Shiga toxin (stx) producing Escherichia coli (STEC) causes various clinical signs in animal and human. In this study, 255 fecal samples from calves showing diarrhea were collected from cattle farms in Chonnam province during the period from January 2005 to July 2005. Twenty six STEC (10%) were isolated from 255 fecal samples by PCR. The isolates displayed three different stx combinations (stx1 [69%], stx1 and stx2 [15%], and stx2 [38%]). The isolates were further studied for virulence associated genes and antimicrobial resistance to define the virulence properties. Intimin (eaeA), enterohemolysin (hlyA), and lipopolysaccharide (rfbE) virulence genes were detected in 6 (23%), 7 (26%), and 1 (3.8%) of the isolates, respectively, by PCR. One isolate possessing rfbE gene was typed as E. coli 0157 : H7 by agglutination test with O and H antisera. All 26 isolates showed susceptibility to amikacin (100%) and the majority of isolates showed high susceptibility to gentamicin (88.5%) and chloramphenicol (73.1%). But all isolates were resistant to penicillin. These results may provide the basic knowledge to establish strategies for the treatment and prevention of enteric disease in calves.

Screening of Anti-Adhesion Agents for Pathogenic Escherichia coli O157:H7 by Targeting the GrlA Activator

  • Sin Young Hong;Byoung Sik Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.329-338
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    • 2023
  • Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that produces attaching and effacing lesions on the large intestine and causes hemorrhagic colitis. It is primarily transmitted through the consumption of contaminated meat or fresh produce. Similar to other bacterial pathogens, antibiotic resistance is of concern for EHEC. Furthermore, since the production of Shiga toxin by this pathogen is enhanced after antibiotic treatment, alternative agents that control EHEC are necessary. This study aimed to discover alternative treatments that target virulence factors and reduce EHEC toxicity. The locus of enterocyte effacement (LEE) is essential for EHEC attachment to host cells and virulence, and most of the LEE genes are positively regulated by the transcriptional regulator, Ler. GrlA protein, a transcriptional activator of ler, is thus a potential target for virulence inhibitors of EHEC. To identify the GrlA inhibitors, an in vivo high-throughput screening (HTS) system consisting of a GrlA-expressing plasmid and a reporter plasmid was constructed. Since the reporter luminescence gene was fused to the ler promoter, the bioluminescence would decrease if inhibitors affected the GrlA. By screening 8,201 compounds from the Korea Chemical Bank, we identified a novel GrlA inhibitor named Grlactin [3-[(2,4-dichlorophenoxy)methyl]-4-(3-methylbut-2-en-1-yl)-4,5-dihydro-1,2,4-oxadiazol-5-one], which suppresses the expression of LEE genes. Grlactin significantly diminished the adhesion of EHEC strain EDL933 to human epithelial cells without inhibiting bacterial growth. These findings suggest that the developed screening system was effective at identifying GrlA inhibitors, and Grlactin has potential for use as a novel anti-adhesion agent for EHEC while reducing the incidence of resistance.

Effects of Fragmentation on the Bird Community in Agricultural Landscapes (농촌 경관에서 파편화가 조류 군집에 미치는 영향)

  • 박찬열;이우신
    • Korean Journal of Environment and Ecology
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    • v.16 no.1
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    • pp.22-33
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    • 2002
  • This study was conducted to investigate the effects of fragmentation on the bird community in Seongduck-ri and Hanggum-ri agricultural landscapes of Kangha-myon, Yangpyong-gun, Kyonggido. We analyzed the composition of tree species, forest environmental structures, structures of agricultural landscapes, avifaunas and guild structures. Number and density of tree species ranging from 6 to 10cm in D.B.H. were high in Seongduck-ri, but basal area was high in Hanggum-ri. Coverages of foliage layers under two meters and over 12 meters were high in Hanggum-ri. Meanwhile, Seongduck-ri agricultural landscapes consisted of twelve patches, dominated by Larix leptolepsis and Robinia pseudoacacia Hanggum-ri landscapes was composed of eleven patches, and dominated by Pinus rigida, Castanea crenate, Larix leptolepsis and paddy field. Dominant bird species showed the same order in two study areas, thirty-six species of birds were observed in Hanggum-ri landscapes, but twenty species were in Seongduck-ri landscapes. Number of species and density belonging to hole-, bush-, ground-nesting and water-, bush-, ground-foraging guild were high in Hanggum-ri landscapes. Number of species and density belonging to hole-nesting guild showed the high value in Hanggum-ri landscapes that was not fragmented. It could be attributed to the low human disturbance and relative high density of medium and large trees in D.B.H. Also, number of species and density belonging to bush-, ground- and water- foraging guild showed the high value in Hanggum-ri landscapes that had the high foliage amounts in the foliage layer under two meters. It will be related with the paddy field, dry field and brook, which patches covered the 28.4% of Hanggum-ri landscapes. Therefore, road not only dominate the heterogeneous patch in agricultural landscapes, but also easily cause the human disturbance. Thus, road construction would decrease the natural patch diversity and edge effects that are important for sustaining the diverse avifauna in agricultural landscapes.

Rhus Verniciflua Stokes Extract Suppresses Expression of Metalloproteinases, iNOS and COX-2 in THP-1 Cells Via Inhibiting NF-𝜅B and MAPK Phosphorylation

  • Ko, Hwanjoo;Jang, Eungyeong;Kim, Youngchul
    • The Journal of Korean Medicine
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    • v.41 no.4
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    • pp.12-26
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    • 2020
  • Objectives: The aim of this study is to investigate the mechanisms involved in the anti-inflammatory and anti-tumor effects of Rhus verniciflua Stokes (RVS) on PMA-differentiated human monocytic leukemia THP-1 cells. Methods: Cells were treated with various concentrations of RVS decoction (0-300㎍/ml) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay. The expressions of MMP-2, MMP-9, TIMP-1, TIMP-2, iNOS and COX-2 mRNA and proteins were measured using RT-PCR and western blotting, respectively. Results: RVS suppressed expression of MMP-2 and MMP-9 mRNA. It also down-regulated iNOS and COX-2 mRNA and protein expression. RVS inhibited NF-𝜅B p65 activity and the phosphorylation of Akt and MAPK (ERK and p38 MAPK). Instead, the phosphorylation of JNK is increased at a very low concentration but decreased at higher concentrations. Conclusion: RVS is regarded to inhibit the expression of MMP and TIMP as well as iNOS and COX-2 gene expression via directly inhibiting the activation of NF-𝜅B and phosphorylation of MAPK pathway in THP-1 cells. This suggests RVS have potential to be used as a therapeutic agent for acute myeloid leukemia (AML).

Preparation and Properties of Phytosphingosine Ascorbate with Retaining Skin Development Effects (피부 활성을 갖는 Phytosphingosine Ascorbate의 합성)

  • Min, Seok-Kee;Jin, Yong-Hoon;Park, Woo-Jung;Eom, Sang-Yong;Kim, Jong-Heon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.167-172
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    • 2004
  • In the human skin, vitamin C (L -ascorbic acid) that is well known as the activated materials has effects that is skin anti-aging and wrinkle repair by giving impetus to collagen biosynthesis and anti-oxidation, and that is the sun screen, a wound recovering, inhibition melanogenesis and so on. In spite of its effects, vitamin C has the defects of the skin stimulation and easily oxidized instability by water, air, heat and light. For solving their matters, many investigation is advanced and its results are synthesized the various vitamin C derivatives. And yet they have not solved the unstable property of vitamin C and were still insufficient for the comparing with the effect of the pure vitamin C itself. In this study, in order to prepare vitamin C derivative of being improved the stability and to apply vitamin C effect in the skin, we prepared new vitamin C derivative, phytosphingosine ascorbate, by using phytosphingosine, one of sphingolipids, which have a distinguished skin affinity. Phytosphingosine ascorbate can be prepared as the ionic bond between amine group (-NH$_2$) of phytosphingosine and hydroxy group (-OH) of vitamin C by way of the relatively simple reaction. So the structure and properties of the synthesized phytosphingosine ascorbate was confirmed the use of elemental analysis (C 58.3 : H 9.3 : N 2.8 : O 29.5), MALDI TOF-MS (Mw=492.58), Ultraviolet spectra (268.5nm), lH NMR, FT-IR spectra, thermal analysis (m.p=l54$^{\circ}C$), HPLC and so on. And we could confirm the anti-bacterial and anti-oxidation effects. Based on these results, we could confirm to prepare a new material that was expected of both effects of vitamin C and phytosphingosine and that is improved properties of vitamin C.

Determination of ibuprofen and its metabolites in human urine by GC-MS (GC-MS에 의한 소변 중 Ibuprofen의 대사체 규명 및 대사 연구)

  • Yu, Dae-Hyung;Cho, Jung-Hum;Hong, Jong-Ki
    • Analytical Science and Technology
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    • v.23 no.2
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    • pp.179-186
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    • 2010
  • The oxidative metabolism of ibuprofen in healthy male urine collected at 3, 6, 9, 12 and 15 h after oral administration of ibuprofen was studied by GC/MS assay. To detect conjugated metabolites of ibuprofen, urine sample was acid-hydrolyzed with 6 M HCl at $100^{\circ}C$ for 30 min. To effectively extract ibuprofen and its metabolites, liquid-liquid extraction (LLE) was conducted at pH 3, 5, and 7, respectively. As a result, LLE at pH 3 was shown to be the best extraction condition. For the determination of trace amounts of ibuprofen and its metabolites in extract, trimethylsilylation (TMS) with BSTFA was applied and followed by GC/MS analysis. In this study, main 5 metabolites including parent drug were detected and these metabolites were assigned as three hydroxylated forms and one carboxylated form. Each metabolite was tentatively identified by both interpretation of mass spectrum and comparison with previously reported results. In addition, time profile of urinary excretion rate for parent drugs and metabolites was studied. Finally, the metabolic pathways of ibuprofen were suggested on the basis of the structural elucidation of its metabolites and excretion profiles.