• Title/Summary/Keyword: Gel electrophoresis

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AN EXPERIMENTAL STUDY ON THE BONE INDUCTION CAPACITY OF THE PORCINE BONE MATRIX-DERIVED BONE MORPHOGENETIC PROTEIN (돼지의 골기질유도 골형성단백질의 골유도능에 관한 실험적 연구)

  • Park, Young-Wook;Lee, Jong-Ho;Kim, Soo-Kyeong
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.19 no.3
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    • pp.265-286
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    • 1997
  • Bone morphogenetic proteins(BMPs) are a group of transforming growth factor beta(TGF-${\beta}$)-related factors and multifunctional proteins, especially the only known biologic factors capable of inducing endochondral bone formation at an extraskeletal site. This study was performed to investigate the effect of the partially purified porcine BMP(pBMP) at an ectopic site. PBMP was partially purified from porcine bone matrix and its activity was monitored by an in vivo bioassay. The purification method utilized extraction of the bone-inducing activity with 4M guanidine, followed by chromatography on heparin-Sepharose. Active fractions were assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. And the fractions were reconstituted with inactive insoluble collagenous bone matrix from rats, acid soluble type I collagen from rat tail and chondroitin-6-sulfate sodium salt and implanted into the pectroralis muscle pouches of Sprague-Dawley rats. And the carrier complex was implanted on the opposite side as control. The rats were sacrificed at the day of 1st, 3rd, 5th, 7th, 11th, 14th and 21st after implantation and examined histologically, radiologically and biochemically. And alkaline phosphatase activity and calcium content were used as indices of bone formation. The results were as follows ; 1. Active fractions were localized in a zone between 31 and 40 KDa on SDS-PAGE. 2. The implanted 3.0mg of the partially purified pBMP induced cartilage and bone in the muscle tissue of rats through an endochondral ossification process. 3. Inactive insoluble bone matrix, type I collagen and chondroitin-6-sulfate have functioned as carriers for pBMP, but revealed some foreign body reactions. 4. Soft X-ray didn't reveal significant change between the experimental and the control group. 5. The alkaline phosphatase activities in the experimental group of 5th, 7th, 11th, 14th and 21st were increased significantly compared with control (p<0.01) with the peak in the group of 11th day. 6. With time, the calcium content of the experimental group increased. And the calcium contents in the experimental group of 11th, 14th and 21st were increased significantly compared with control (p<0.01).

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Studies of Insecticide Resistance in Green Peach Aphids, Myzus persicae(Sulz) III. Acephate Resistance, Cross-Resistance, and Esterase Isozymes (복숭아혹진딧물의 살충제 저항성(抵抗性)에 관한 연구(硏究) III. Acephate저항성(抵抗性) 발달(發達), 교차저항성(交叉抵抗性) 및 Esterase Isozymes)

  • Choi, Seung-Yoon;Kim, Gil-Hah
    • Korean journal of applied entomology
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    • v.25 no.2 s.67
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    • pp.99-105
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    • 1986
  • The green peach aphids(Myzus persicae) collected in a field had been successively selected by acephate(O, S-dimethyl N-acetyl phosphoroamidothioate) in the laboratory. The selected aphid strain in the 20th generation demonstrated relatively high resistance to acephate as well as relatively high cross-resistance to cypermethrin and oxydemeton-methyl, except pirimicarb. The different esterase isozymes with the strains were detected by the agarose gel electrophoresis and among the isozymes the band of ${\beta}-2$ was only specific for the acephate resistant strains.

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Separation of Egg White Using HPLC with Change of Mobile Phase and Temperature (HPLC에서 이동상 변화와 온도에 따른 난백의 분리)

  • Do, Jin-Sun;Song, Shin-Young;Cho, Ki-Jung;Kim, In-Ho
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.829-834
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    • 2011
  • Lysozyme in egg white functions as bacteriolysis agent and ovalbumin plays a role as antigen in immune system. Egg white analysis methods usually include electrophoresis, gel permeation chromatography and reversed-phase HPLC(RP-HPLC). Among them, RP-HPLC was selected for rapid analysis and C18 column(Agilent, USA) was used as HPLC column. Optimum conditions were searched by changing mobile phase and temperature. Capacity factor and resolution were calculated and compared for various elution conditions. In the isocratic elution, mobile phase volume ratio was changed from 30/70/0.1 to 60/40/0.1(Acetonitrile(ACN)/Distilled water(DW)/Trifluoroacetic acid(TFA)). ACN composition was increased by 10% and temperature was set as $20^{\circ}C$. In the gradient elution, ACN/DW ratio was changed from 10/90 to 60/40 during 20 minute and temperature was varied as 20, 30 and $40^{\circ}C$. In the isocratic elution, three peaks were separated at 50/50/0.1. Lysozyme and ovalbumin were confirmed as first and third peak in three peaks respectively. In the gradient elution, four peaks were separated at $30^{\circ}C$. Lysozyme and ovalbumin were confirmed as first peak and third peak in four peaks respectively.

Assessing Water Quality of Siheung Stream in Shihwa Industrial Complex Using Both Principal Component Analysis and Multi-Dimensional Scaling Analysis of Korean Water Quality Index and Microbial Community Data (Principal Component Analysis와 Multi-Dimensional Scaling 분석을 이용한 시화공단 시흥천의 수질지표 및 미생물 군집 분포 연구)

  • Seo, Kyeong-Jin;Kim, Ju-Mi;Kim, Min-Jung;Kim, Seong-Keun;Lee, Ji-Eun;Kim, In-Young;Zoh, Kyung-Duk;Ko, Gwang-Pyo
    • Journal of Environmental Health Sciences
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    • v.35 no.6
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    • pp.517-525
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    • 2009
  • The water quality of Lake Shihwa had been rapidly deteriorating since 1994 due to wastewater input from the watersheds, limited water circulation and the lack of a wastewater treatment policy. In 2000, the government decided to open the tidal embankment and make a comprehensive management plan to improve the water quality, especially inflowing stream water around Shihwa and Banwol industrial complex. However, the water quality and microbial community have not as yet been fully evaluated. The purpose of this study is to investigate the influent water quality around the industrial area based on chemical and biological analysis, and collected surface water sample from the Siheung Stream, up-stream to down-stream through the industrial complex, Samples were collected in July 2009. The results show that the downstream site near the industrial complex had higher concentrations of heavy metals (Cu, Mn, Fe, Mg, and Zn) and organic matter than upstream sites. A combination of DGGE (Denaturing Gradient Gel Electrophoresis) gels, lists of K-WQI (Korean Water Quality Index), cluster analysis, MDS (Multi-Dimensional Scaling) and PCA (Principal Component Analysis) has demonstrated clear clustering between Siheung stream 3 and 4 and with a high similarity and detected metal reducing bacteria (Shewanella spp.) and biodegrading bacteria (Acinetobacter spp.). These results suggest that use of both chemical and microbiological marker would be useful to fully evaluate the water quality.

Antioxidant Activity and Radioprotection of Two Flavonoids from Propolis (프로폴리스에서 분리한 플라보노이드 화합물의 항산화 활성 및 방사선 방어효과)

  • Jeong, Ill-Yun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.162-166
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    • 2005
  • Two flavonoids, 7-O-methyl-3',4'-didehydroxy quercetin (MDQ) and quercetin, isolated from Chinese propolis, which is the generic name for the resinous substance collected by honeybees from various plant sources, were tested for their antioxidant activity and protective effect against radiation-induced DNA damage in mouse lymphocytes. In antioxidant test, both compounds provided a dose-dependent scavenging effect on DPPH radical and a dose-dependent inhibitory effect on lipid peroxidation in mouse liver. Quercetin showed stronger scavenging and inhibitory effect than MDQ, and it also provided strong inhibition on superoxide anion radical generated in xanthine-xanthine oxidase system, but there was no inhibitory ability for MDQ. In comet assay using single cell gel electrophoresis, MDQ and quercetin showed a protective effect against DNA damage caused by gamma irradiation. They reduced DNA damage to 54% (p<0.01) and 53% (p<0.01) at 25 $\mu$mol, respectively. These results suggest that free radical scavenging seems to be associated with their catechol form on the B ring, and radioprotection appears to be a likely mechanism of antioxidant activity by these flavonoids.

Protein Composition and Biological Activities of Bombus ignitus Venom (호박벌 일벌독의 성분 분석 및 생리활성 탐색)

  • Han, Sang-Mi;Lee, Kwang-Gill;Yeo, Joo-Hong;Kweon, Hae-Yong;Woo, Soon-Ok;Yoon, Hyung-Joo;Kim, Me-Ae;Kim, Won-Tae;Baek, Ha-Ju
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.229-234
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    • 2007
  • Pure Bombus ignitus venom samples were submitted to two-dimensional gel electrophoresis. A total of 64 excised spots were analyzed by mass spectrometry. Three main proteins resulted in the identification have not been described in other bee venoms before. Dose-dependence against human carcinoma (Hep3B, BT-20, A549 and AGS) were observed from 1ng/ml to 100ng/ml. Expecially, the treatment of 100ng/ml B. ignitus venoms showed the highest cytotoxicity with 55% against hepatocellular carcinoma (Hep3B). The B. ignitus venoms showed strong antimicrobial activities against Enterococcus faecium and Shigella sonnei, and practically antimicrobial activity against the other microorganisms tested. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of E. faecium and S. sonnei, were 0.256ug/ml, respectively.

No Associations between Schizophrenia and D22S280 Marker on Synapsin III Gene in Korean Males (한국인 남자에서 Synapsin III 유전자의 D22S280 표지자와 정신분열병의 연합연구)

  • Lee, Yu-Sang;Park, Chong-Won;Lee, Seung-Yeoun;Lee, Suk-Jin;Park, Yong-Bum;Shin, Yoon-Sik;Yoo, Jang-Keun;Hong, Kyung Sue;Yang, Byung-Hwan
    • Korean Journal of Biological Psychiatry
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    • v.13 no.4
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    • pp.260-266
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    • 2006
  • Objectives : Synapsin III near VCFS region on chromosome 22q affects. It could be an interesting candidate gene for schizophrenia. D22S280 is a highly polymorphic genetic marker residing in synapsin III. We examined association of D22S280 marker on synapsin III with Korean patients with schizophrenia. Methods : The subjects were 46 male Korean patients with schizophrenia and 60 male normal controls. Using polymerase chain reaction, gel electrophoresis, ABI 310 genetic analyzer, and GeneScan Collection 3.1 software, we confirmed genotypes of D22S280 marker. We examined Hardy-Weinberg equilibrium and case-control association using SAS/Genetic 9.1.3. Results : Genotypes of both schizophrenia and control groups were in Hardy-Weinberg equilibrium. We could not find any significant statistical differences in allele-wise(${\chi}^2$=10.4, df=6, p=0.098) and genotype-wise (${\chi}^2$=22.1 df=19, p=0.258) analyses of D22S280 marker between schizophrenia and normal controls. Individual allele analyses with df=1 showed significant differences in A1(p=0.025) and A7(p=0.034) allele, which were not significant following Bonferroni corrections(A1:p=0.177, A7:p=0.235). Conclusion : We couldn't find any association between schizophrenia and the synapsin III gene. Given the small number of subjects studied, further investigations are needed.

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A brief method for preparation of gintonin-enriched fraction from ginseng

  • Choi, Sun-Hye;Jung, Seok-Won;Kim, Hyun-Sook;Kim, Hyeon-Joong;Lee, Byung-Hwan;Kim, Joon Yong;Kim, Jung-Hyun;Hwang, Sung Hee;Rhim, Hyewon;Kim, Hyoung-Chun;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.39 no.4
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    • pp.398-405
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    • 2015
  • Background: Ginseng has been used as a tonic for invigoration of the human body. In a previous report, we identified a novel candidate responsible for the tonic role of ginseng, designated gintonin. Gintonin induces $[Ca^{2+}]_i$ transient in animal cells via lysophosphatidic acid receptor activation. Gintonin-mediated $[Ca^{2+}]_i$ transient is linked to anti-Alzheimer's activity in transgenic Alzheimer's disease animal model. The previous method for gintonin preparation included multiple steps. The aim of this study is to develop a simple method of gintonin fraction with a high yield. Methods: We developed a brief method to obtain gintonin using ethanol and water. We extracted ginseng with fermentation ethanol and fractionated the extract with water to obtain water-soluble and water-insoluble fractions. The water-insoluble precipitate, rather than the water-soluble supernatant, induced a large $[Ca^{2+}]_i$ transient in primary astrocytes. We designated this fraction as gintonin-enriched fraction (GEF). Results: The yield of GEF was approximately 6-fold higher than that obtained in the previous gintonin preparation method. The apparent molecular weight of GEF, determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was equivalent to that obtained in the previous gintonin preparation method. GEF induced $[Ca^{2+}]_i$ transient in cortical astrocytes. The effective dose (ED50) was $0.3{\pm}0.09{\mu}g/mL$. GEF used the same signal transduction pathway as gintonin during $[Ca^{2+}]_i$ transient induction in mouse cortical astrocytes. Conclusion: Because GEF can be prepared through water precipitation of ginseng ethanol extract and is easily reproducible with high yield, it could be commercially utilized for the development of gintoninderived functional health food and natural medicine.

Differential Expression of Glucose Transporter Gene in Mouse Early Embryos (생쥐 초기배아의 Glucose Transporter유전자 발현 양상에 관한 연구)

  • Youm, Hye-Won;Byun, Hye-Kyung;Song, Gyun-Ji;Kim, Hae-Kwon;Lee, Ho-Joon
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.77-86
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    • 1998
  • The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture ($93.8{\pm}3.1$, n=35) is significantly higher than that of control and glucose-free group ($76.6{\pm}3.8$, n=35 and $68.2{\pm}4.3$, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.

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Lysosomal Acid Phosphatase in Regenerating Salamander Limbs Studied with Monoclonal Antibodies (리소솜 Acid Phosphatase에 대한 단일 항체를 이용한 도롱뇽 다리 재생 연구)

  • 주봉건;박상렬;최의열;김원선
    • The Korean Journal of Zoology
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    • v.39 no.4
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    • pp.426-436
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    • 1996
  • In previous studies, we have shown that lysosomal add phosphatase (LAP) activity increases at the dedifferentiation stage in the regenerating larval limbs of salamander, Hynobius leechii. Monoclonal antibodies against LAP were generated to determIne the spatial and temporal distribution of the protein In the regenerates.A total of 22 monoclonal antihodies recognizIng different epftopes of the protein were obtained, of which five strongly stained the regenerating limb by imunohistochemistry. in LAP immunohistochemical examination, LAP showed distribution coincident with the state of dedifferentiation, both spatially and temporally, in the limb regenerates. When unfractioned protein of regenerating salamander limbs were separated by gel electrophoresis and immunoblotted, the antibodies recognized a single protein band of 53 kl)a, which comigrates with a monomerlc subunit of IAR Using the anti-IAP antibodIes as probe, we investigated the cross-reactivities of LAPs from other sources. The immunoreadive bands on Western blots appeared to be the same In molecular mass-53 kl)a in axoloti and Xenopus, but no protein band was detected in mouse, Drosophila, or C. elegans.These results show that the antibodies generated in this study spedfically recognize Hynoblus leeclili IAp and that IAPs may be highiy conserved among amphibians. Furthermore, the distdbution of the protein is consistent with a role for LAP in the dedifferentiation process of limb regeneration.

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