• Title/Summary/Keyword: Structural activity

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Clonig of CM-cellulase Gene of Rhizobium meliloti TAL1372 in Escherichia coli (Rhizobium meliloti TAL1372에서 섬유소분해효소 유전자 클로닝)

  • Park, Yong-Woo;Lim, Sun-Teak;Kang, Kyu-Young;Yun, Han-Dae
    • Applied Biological Chemistry
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    • v.38 no.4
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    • pp.313-319
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    • 1995
  • The involvement of the cell-wall degrading enzymes in Rhizobium has long been an unsolved question about the infection process in the formation of root nodule. To assess the contribution of the cellulase to the nodulation of rhzobia, here we report the production of cellulase from R. meliloti TAL1372 which degrade carboxymethylcellulose (CMC) model substrate with CMC-plate method. We constructed a genomic library by cloning Sau3A-digested genomic DNA from R. meliloti TAL1372 into the BamHI site of the cosmid vector pLAFR3. Out of more than one thousand transductants of E. coli, one clone (pRC8-71) had CM-cellulase activity and contained pLAFR3 cosmid with 30 kb insert of R. meliloti DNA The product of CM-cellulase gene was analyzed by native PAGE. About 45 kD protein was considered to be a product of the gene. Tn5 mutagenesis reveals that the structural gene located in a ca. 3 kb KpnI fragment. The cellulase-minus mutants of R. meliloti TAL1372 were obtained by Tn5 mutagenesis of pRC8-71 and marker exchange techniques. Analyses of the nodulation ability of these Tn5 mutants showed that the CM-cellulase gene of R. meliloti TAL1372 may be involved in early nodulation development on alfalfa (Medicago satiua).

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Isolation and Characterization of Bacterial Cellulose-Producing Bacteria for Silver Nanoparticle Synthesis (은 나노입자 합성을 위한 Bacterial Cellulose 생산 세균의 분리 및 특성)

  • Yoo, Ji-Yeon;Jang, Eun-Young;Son, Yong-Jun;Park, Soo-Yeun;Son, Hong-Joo
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.120-126
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    • 2018
  • As a basic study for environment-friendly production of bacterial cellulose (BC) dressing with antimicrobial activity, we isolated and identified acetic acid bacteria which are resistant to silver ions and can biosynthesize silver nanoparticles. Furthermore, conditions of BC production by selected strain were also investigated. Strain G7 isolated from decayed grape skin was able to grow in the presence of 0.1 mM $AgNO_3$ which was identified as Acetobacter intermedius based on 16S rRNA gene analysis. BC production was the highest in a medium containing 2% glucose as a carbon source, 2% yeast extract as a nitrogen source, and 0.115% acetic acid as a cosubstrate. Structural properties of BC produced in optimal medium were studied using Fourier-transform infrared spectroscopy and X-ray diffractometer, and it was found that BC produced was cellulose type I that was the same as a typical native cellulose. When strain G7 was cultured in an optimal medium containing 0.1 mM $AgNO_3$, the color of the culture broth turned into reddish brown, indicating that silver nanoparticles were formed. As a result of UV-Vis spectral analysis of the culture, it was found that a unique absorption spectrum of silver nanoparticles at 425 nm was also observed. Scanning electron microscopic observations showed that silver nanoparticles were formed on the surface and pores of BC membrane.

Protective Effects of Flavonoids from the Boehmeria quelpaertense against H2O2-Induced Cytotoxicity in H9c2 Cardiomyoblast Cells (H9c2 심근세포에서 제주모시풀(Boehmeria quelpaertense)로부터 분리된 flavonoids의 H2O2로 유도된 독성 보호 효과)

  • Woo, Kyeong-Wan;Sim, Mi-Ok;Bak, Ho;Jung, Ho Kyung;An, Byeongkwan;Ham, Seong-Ho;Park, Jong Hyuk;Cho, Hyun-Woo
    • Korean Journal of Plant Resources
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    • v.31 no.1
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    • pp.1-9
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    • 2018
  • As a part of an infrastructure project on medicinal herb-based remedies, we conducted a phytohemical investigation of the 100% MeOH extract from the aerial part of Boehmeria quelpaertense; our findings resulted in the isolation of flavonoids (1-2), isoquercitrin (1) and hyperoside (2). The identification and structural elucidation of these compounds were based on $^1H$-, $^{13}C-NMR$, and LC ESI IT-TOF MS data. All the compounds isolated from this plant were reported for the first time. In this study, we examined the antioxidant activity of the 1 and 2 on the hydrogen peroxide ($H_2O_2$)-induced oxidative stress in a Rat Cardiomyoblast cell line (H9c2). The pretreatment of the flavonoids showed that it protects against $H_2O_2$-mediated cell death in the H9c2 cell line. Also, it decreases the intracellular reactive oxygen species (ROS) levels by the flavonoids in the $H_2O_2$-treated H9c2 cell line. These results showed that the 1 and 2 are a source of antioxidants. As a result, they might be helpful in preventing the progress of various oxidative stress mediated diseases, including myocardial infarction.

Properties of Chemically Activated MSWI(Municipal Solid Waste Incinerator) Mortar (도시 폐기물 소각재를 이용한 화학적 활성화 모르타르의 특성)

  • Jo, Byung-Wan;Kim, Kwang-Il;Park, Jong-Chil;Park, Seung-Kook
    • Journal of the Korea Concrete Institute
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    • v.18 no.5 s.95
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    • pp.589-594
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    • 2006
  • The recycling of industrial wastes in the concrete manufacturing is of increasing interest worldwide, due to the high environmental impact of the cement and concrete industries and to the rising demand of infrastructures, both in industrialized and developing countries. The production of municipal wastes in the South Korea is estimated at about 49,902 ton per day and only 14.5% of these are incinerated and principally disposed of in landfill. These quantities will increase considerably with the growth of municipal waste production, the progressive closing of landfill, so the disposal of municipal solid waste incinerator(MSWI) ashes has become a continuous and significant issue facing society, both environmentally and economically. MSWI ash is the residue from waste combustion processes at temperature between $850^{\circ}C\;and\;1,000^{\circ}C$. And the main components of MSWI ash are $SiO_2,\;CaO\;and\;Al_2O_3$. The aim of this study is to find a way to useful application of MSWI ash(after treatment) as a structural material and to investigates the hydraulic activity, compressive strength development composition variation of such alkali-activated MSWI ashes concrete. And it was found that early cement hydration, followed by the breakdown and dissolving of the MSWI-ashes, enhanced the formation of calcium silicate hydrates(C-S-H). The XRD and SEM-EDS results indicate that, both the hydration degree and strength development are closely connected with a curing condition and a alkali-activator. Compressive strengths with values in the 40.5 MPa were obtained after curing the activated MSWI ashes with NaOH+water glass at $90^{\circ}C$.

Testing the Biobehavioral Family Model in Understanding the Eating Problems of Adolescent Girls (여고생의 섭식문제 구조모형 구축: 생체행동가족모형의 적용)

  • Park, Ji-Young;Baek, Su-Yon;Kim, Hee-Soon;Lim, Jung-Ha;Kim, Tae-Hyung
    • Child Health Nursing Research
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    • v.19 no.3
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    • pp.228-237
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    • 2013
  • Purpose: This study was done to test a hypothesized model, the Biobehavioral Family Model (BBFM), on the relationship of family emotional climate, security of parent-child relationship, depression symptoms and eating problems in adolescent girls, to further understanding of eating problems in this population. Methods: With a convenience sample of 647 girls, aged 15 to 18, a self-report survey was conducted which included the Korean form of the Eating Attitude Test (EAT-26) to assess eating problems. Results: The estimated results of the structural equation modeling indicated a good fit of data to the hypothesized model proposing that family emotional climate and security of parent-child relationship were associated with the risk of eating problems by way of depression symptoms. That is, negative family emotional climate and insecure parent-child relationship increased the risk of eating problems indirectly by way of depression symptoms. Conclusion: The findings are consistent with the BBFM, which suggests a psychobiologic influence of specific family processes on children's stress-sensitive physical disease activity by way of depression symptoms. Therefore, the applicability of the BBFM for understanding adolescent girls' eating problems is supported. The psychobiologic pathways from depression to eating pathology should be addressed in future studies.

Molecular Cloning and Characterization of a P38-Like Mitogen-Activated Protein Kinase from Echinococcus granulosus

  • Lu, Guodong;Li, Jing;Zhang, Chuanshan;Li, Liang;Bi, Xiaojuan;Li, Chaowang;Fan, Jinliang;Lu, Xiaomei;Vuitton, Dominique A.;Wen, Hao;Lin, Renyong
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.759-768
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    • 2016
  • Cystic echinococcosis (CE) treatment urgently requires a novel drug. The p38 mitogen-activated protein kinases (MAPKs) are a family of Ser/Thr protein kinases, but still have to be characterized in Echinococcus granulosus. We identified a 1,107 bp cDNA encoding a 368 amino acid MAPK protein (Egp38) in E. granulosus. Egp38 exhibits 2 distinguishing features of p38-like kinases: a highly conserved T-X-Y motif and an activation loop segment. Structural homology modeling indicated a conserved structure among Egp38, EmMPK2, and H. sapiens $p38{\alpha}$, implying a common binding mechanism for the ligand domain and downstream signal transduction processing similar to that described for $p38{\alpha}$. Egp38 and its phosphorylated form are expressed in the E. granulosus larval stages vesicle and protoscolices during intermediate host infection of an intermediate host. Treatment of in vitro cultivated protoscolices with the p38-MAPK inhibitor ML3403 effectively suppressed Egp38 activity and led to significant protoscolices death within 5 days. Treatment of in vitro-cultivated protoscolices with $TGF-{\beta}1$ effectively induced Egp38 phosphorylation. In summary, the MAPK, Egp38, was identified in E. granulosus, as an anti-CE drug target and participates in the interplay between the host and E. granulosus via human $TGF-{\beta}1$.

Prediction Model of Fatigue in Women with Rheumatoid Arthritis (여성 류마티스 관절염 환자의 피로 예측 모형)

  • Lee, Kyung-Sook;Lee, Eun-Ok
    • Journal of muscle and joint health
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    • v.8 no.1
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    • pp.27-50
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    • 2001
  • Rheumatoid arthritis is a chronic systemic autoimmune disease. Although the joints are the major loci of the disease activity, fatigue is a common extraarticular symptom that exists in all gradations of rheumatoid arthritis. Fatigue is defined as a subjective sense of generalized tiredness or exhaustion and has multiple dimensions. Therefore fatigue is a common and frequent problem for those with rheumatoid arthritis. In fact, 88-100% of individuals with rheumatoid arthritis experience fatigue. Especially the degree of fatigue is higher in women than men with rheumatoid arthritis. Despite the importance of fatigue among the patients with rheumatoid arthritis, the mechanism that leads to fatigue in rheumatoid arthritis is not completely understood. This study was intended to test and validate a model to predict fatigue in women with rheumatoid arthritis. Especially it was intended to identify the direct and indirect effects of the variables of pain, disability, depression, sleep disturbance, morning stiffness, and symptom duration to fatigue. Data were collected by questionnaires including Multidimensional Assesment of Fatigue(Tack, 1991), numeric scale of pain, graphic scale of joints, Ritchie Articular Index, Korean Health Assessment Questionnaire(Bae, et al., 1998), Inventory of Function Status(Tulman, et al., 1991), Center for Epidemiologic Studies-Depression, and Korean Sleep Scale(Oh, et al 1998). The sample consisted of 345 women with a mean duration of rheumatoid arthritis for 10.06 years and a mean age of 49.64 years. SPSS win and Win LISREL were used for the data analysis. Structural equation modeling revealed the overall fit of the model. Pain predicted fatigue directly and indirectly through disability, depression, and sleep disturbance. Disability, sleep disturbance predicted fatigue only directly, while depression only indirectly through disability and sleep disturbance. Also morning stiffness and symptom duration predicted fatigue through disability and depression. All predictors accounted for 65% of the variance of fatigue. Depression, pain, and disability predicted sleep disturbance. Depression had reciprocal relationship with disability and they both were predicted by pain directly and indirectly. In summary, pain, depression, disability, sleep disturbance, morning stiffness, and symptom duration contributed to the fatigue of patients with rheumatoid arthritis. The best predictor of fatigue was pain. This finding indicates that the modification of pain, depression, disability, sleep disturbance, morning stiffness could be nursing intervention for relief or prevention of fatigue.

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Functions of a-Tropomyosin Are Mainly Dependent upon the Local Structures of the Amino Terminus (a-Tropomyosin의 아미노 말단 구조가 기능에 미치는 영향)

  • Cho, Young-Joon
    • Journal of Life Science
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    • v.14 no.5
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    • pp.770-777
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    • 2004
  • It has been previously reported that unacetylated a-tropomyosin(TM) produced in E. coli failed to bind to actin while acetylated muscle TM and Ala-Ser dipeptide fusion TM (AS-TM) bound well to actin. In order to determine the structural requirement of the amino terminus for high actin affinity, a recombinant tropomyosin (Ala-TM) that a single Ala residue was added to the amino terminus of Ala-TM was constructed, overexpressed, and purified from E. coli. Actin affinity of Ala-TM was 2.3$\times$$10^{6}$$M^{-1}$, whereas that of unacetylated TM was considerably lower than 0.1$\times$$10^{-6}$$M^{-1}$ indicating that addition of a single Ala residue to the amino terminus drastically increased, at least twenty times, actin affinity of TM. Ala-TM, however, bound to actin about three times weaker than acetylated TM and AS- TM, implying that the addition of an Ala residue was insufficient for complete restoration of high actin affinity. While Ala-TM, AS-TM, and muscle TM showed inhibition and activation of actomyosin Sl ATPase activity depending on myosin Sl concentration, the degree of inhibition and activation was different from each other. AS-TM exhibited the greatest inhibition of the ATPase at low Sl concentration, whereas the greatest activation of the ATPase was observed with muscle TM. These results, together with previous findings, strongly suggested that local structure of the amino terminus is the crucial functional determinant of TM.

Functional Cardiomyocytes Formation Derived from Parthenogenetic Mouse Embryonic Stem Cells (단위발생 유래 생쥐 배아줄기세포의 기능성 심근세포 형성)

  • Shin, Hyun-Ah;Kim, Eun-Young;Lee, Young-Jae;Lee, Keum-Sil;Park, Eun-Mi;Lee, Hoon-Taek;Chung, Kil-Saeng;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.2
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    • pp.139-147
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    • 2002
  • Objective : This study was to establish a reproducible differentiation system from the parthenogenetic mouse embryonic stem (P-mES02) cells into functional cardiomyocytes like as in vitro fertilization mouse embryonic stem (mES01) cells. Materials and Methods: To induce differentiation, P-mES02 cells were dissociated and aggregated in suspension culture environment for embryoid body (EB) formation. For differentiation into cardiomyocytes, day 4 EBs were treated with 0.75% dimethyl sulfoxide (DMSO) for another 4 days (4-/4+) and then were plated onto gelatin-coated dish. Cultured cells were observed daily using an inverted light microscope to determine the day of contraction onset and total duration of continuous contractile activity for each contracting focus. This frequency was compared with the results of DMSO not treated P-mES02 group (4-/4-) and mES01 groups (4-/4+ or 4-/4-). For confirm the generation of cardiomyocytes, beating cell masses were treated with trypsin-EDTA, dispersed cells were plated onto glass coverslips and incubated for 48 h. Attached cells were fixed using 4% paraformaldehyde and incubated with specific antibodies (Abs) to detect cardiomyocytes (anti-sarcomeric ? -actinin Ab, 1 : 100; anti-cardiac troponin I Ab, 1 : 2000) for 1 h. And the cells were finally treated with FITC or TRITC labelled 2nd Abs, respectively, then they were examined under fluorescence microscopy. Results: Rhythmically contracting areas in mES01 or P-mES02 cells were firstly appeared at 9 or 10 days after EBs plating, respectively. The highest cumulative frequency of beating EBs was not different in both treatment groups (mES01 and P-mES02, 4-/4+) with the results of 61.3 % at 13 days and 69.8% at 15 days, respectively. Also, the contracting duration of individual beating EBs was different from minimal 7 days to maximal 53 days. However, DMSO not treated groups (mES01 and P-mES02, 4-/4-) also had contracting characteristics although their frequency was a few compared to those of DMSO treated groups (6.0% and 4.0%). Cells recovered from the spontaneously contracting areas within EBs in both treated groups were stained positively with muscle specific anti-sarcomeric ? -actinin Ab and cardiac specific anti-cardiac troponin I Ab. Conclusion: This study demonstrated that the P-mES02 cell-derived cardiomyocytes displayed similarly structural properties to mES01 cell-derived cardiomyocytes and that the DMSO treatment enhanced the cardiomyocytes differentiation in vitro.

Nature of contact between the Ogcheon belt and Yeongnam massif and the Pb-Pb age of granitic gneiss in Cheondong-ri, Danyang (단양 천동리 지역 옥천대/영남육괴의접촌관계와 소위 화강암질 편마암의 Pb-Pb 연대)

  • 권성택;이진한;박계헌;전은영
    • The Journal of the Petrological Society of Korea
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    • v.4 no.2
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    • pp.144-152
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    • 1995
  • The Jangsan Quartzite of the Joseon Supergroup and the foliated granite (so-called granitlc gneiss of presumed Precambrian age) of the Yeongnam massif are in direct contact at Cheondong-ri area, 6 km @SE of Danyang. sllthough it has been thought traditionally that the Jangsan Quartzite overlies unconformably the f&ted granite, it is difficult to interpret the contact as an unconformity smce the basal conglomerate in- the lower part of the Jangsan Quartzite does not have any clast of the foliated granite, Rather, recent structural studies of this area indlcate that the contact is a ductile shear zone. However, the sense and age of the shear movement are still problematic. Our mesoscopic and microscopic studies of &tre Cheondong-11 semi-brittle shear zone involving foliated cataclasite and phyllonite, which is a pa& of the Ogdong fault, indlcate a top-to-the northeast shearing, i.e., dextral strike slip. We also performed Pb-Pb dating for the age-unknown foliated granite, since the age of deformed granite ccarr emtrain the maximum age of deformation. The whole rock and feldspar Pb isotape data for the foliated granite and a micaceous xenolith define an isoc chron age of $2.16{\pm}0.15$ Ga ($2{\sigma}$;MSWD=4.4) which is interpreted as the emplacement age of the granite. This early Proterozoic age agrees with those of Precambrian igneous activity In the Yeongnam massif reported previously. The obtaiPrfid gge confirms the traditional idea about the age of the foliated granite and indicates that other methd(s) should be employed to constrain the age of the shear movement.

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