• Title/Summary/Keyword: immunoblotting

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Regulation and Expression of Glycerol-3-phosphate Dehydrogerlase (GPDH) in Drosophila melanogaster (노랑초파리의 $\alpha$-Glycerol-3-phosphate Dehydrogenase (GPDH)의 발현과 조절)

  • 김세재;이정주남궁용김경진
    • The Korean Journal of Zoology
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    • v.34 no.1
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    • pp.123-130
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    • 1991
  • Several parameters of u -glycerol-3-pholphate dehydrogenase (GPDH) such as activity, content and translatable mRNA levels were measured to elucidate mechanism underlving developmental and tissue specific regulation of 6PDH activity in Drosophila melonogastrr. In adult segments, most of total GPDH activity (62%1 Iwas detected in thorax where GPDH-1 resided, while 32% of total GPDH aUiviD was only detected in abdomen where GPDH-3 resided. The relative synthesis of GPDH was, however, similar in both tissues, although 58% of total GPDH was synthesized in abdomen. These results strongly suggest that the turnover rate of the abdominal enzyme (GPDH-3) was much more rapid than that of thoracic enzymes (GPDH-1). In nitro translation and immunoblotting experiments also indicate that GPDH-3 was arised by posttranslational modification from a single polypeptide (GPDH-1).

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Epidermis Proliferative Effect of the Panax ginseng Ginsenoside $Rb_2$

  • Choi, Seong-Won
    • Archives of Pharmacal Research
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    • v.25 no.1
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    • pp.71-76
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    • 2002
  • Ginseng has been used as a traditional medicine with various therapeutic effects. However, it is still unknown which component of this plant is effective at promoting wound healing. Recently, ginsenoside $Rb_2$ has been reported to improve wound healing. In this study, to investigate the reported wound healing effect of the ginsenoside $Rb_2$, cell morphology and protein factors involved in epidermal formation were evaluated by immunshemical and immunoblotting analysis. $Rb_2$ stimulated epidermal cell proliferation, and the cell showed a 1.5-fold increase in thymidine uptake compared to the control (p<0.05, n=3). Futheremore $Rb_2$, was found to stimulate epidermis formation in a dose-dependent manner in raft culture, and to dose dependently enhance the expressions of protein factors related to cell proliferation, namely, epidermal growth factor and its receptor, fibronectin and its receptor, keratin 5/14, and collagenase 1 (p<0.05, n=3~9). It is believed that ginsenoside $Rb_2$, enhances epidermal cell proliferation by upregulating the expressions of these proliferation-related factors.

Exogenous JH and ecdysteroid applications alter initiation of polydnaviral replication in an endoparasitoid wasp, Cotesia plutellae (Braconidae: Hymenoptera)

  • Park, Bok-Ri;Kim, Yong-Gyun
    • BMB Reports
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    • v.44 no.6
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    • pp.393-398
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    • 2011
  • Polydnaviruses are a group of double-stranded DNA viruses and are symbiotically associated with some ichneumonoid wasps. As proviruses, the replication of polydnaviruses occurs in the female reproductive organ at the pupal stage. This study analyzed the effects of two developmental hormones, juvenile hormone (JH) and ecdysteroid, on the viral replication of Cotesia plutellae bracovirus (CpBV). All 23 CpBV segments identified contained a conserved excision/rejoining site ('AGCTTT') from their proviral segments. Using quantitative real-time PCR based on this excision/rejoining site marker, initiation of CpBV replication was determined to have occurred on day 4 on the pupal stage. Pyriproxyfen, a JH agonist, significantly inhibited adult emergence of C. plutellae, whereas RH5992, an ecdysteroid agonist, had no inhibitory effect. Although RH5992 had no effect dose on adult development, it significantly accelerated viral replication. The results of immunoblotting assays against viral coat proteins support the effects of the hormone agonists on viral replication.

Identification of a Fusion-associated Protein in the Skeletal Myoblast Using Monoclonal Antibody (단일클론항체를 이용한 배양 계배 근원세포의 융합과 연관된 단백질의 확인)

  • Kim, Chons-Rak;Won
    • The Korean Journal of Zoology
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    • v.35 no.1
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    • pp.29-36
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    • 1992
  • The present study describes the production of monoclonal antibodies against cultured chick myoblast to pursue critical proteins in muscle cell fusion. Among a panel of monoclonal antibodies, three, Mll-3H 13, Mll-3Hl8 and Mll-3H35 were inhibited movblast fusion. A single 101-kDa antigen reactive with monoclonal antibody Mll-3H35 was detected by radioimmu-noprecipitation or by immunoblotting. During the course of myogenesis, the level of the protein remarkably decreased as the cells there differentiated. These results suggest that the protein platys a direct role in the process of myoblast fusion mechanism.

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Anti-inflammatory and Radical Scavenging Effects of Spirodela polyrrhiza

  • Jeon, Hoon
    • Natural Product Sciences
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    • v.16 no.2
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    • pp.111-115
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    • 2010
  • Anti-inflammatory and anti-oxidant effect of Spirodela polyrrhiza (Lemnaceae), a widely used traditional medicinal plant were investigated. In macrophages nitric oxide (NO) is released as an inflammatory mediator and has been proposed to be an important modulator of many pathophysiological conditions including inflammation. 85% MeOH extracts of S. polyrrhiza (0.01, 0.1, 1 mg/mL) suppressed nitric oxide production in interferone-$\gamma$ (IFN-$\gamma$) and lipopoloysaccharide(LPS)-stimulated macrophages. It also attenuated the expression of inflammatory enzymes like inducible NOS (iNOS) and cyclooxygenase-2(COX-2) as assessed by immunoblotting with specific antibodies. Moreover, the values obtained with DPPH radical, superoxide anion and NO radical scavenging assay showed that S. polyrrhiza has potent antioxidant properties as a natural ROS scavenger. The results of the present study suggest the potential use of S. polyrrhiza in the treatment of ROS-mediated chronic inflammatory diseases such as atherosclerosis and rheumatoid arthritis.

Bacillus cellulyticus K-12 Crystalline Cellulose-Degrading Avicelase Gene and Expression in Eschterichia coli

  • Cheorl-Ho Kim;Woo
    • The Korean Journal of Food And Nutrition
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    • v.6 no.4
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    • pp.314-321
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    • 1993
  • We have cloned the Bacillus cellulyticus K-12 avicelase (Avi, E.C.3.2.1.4) gene (ace A) In E. coli. This was accompanied by using the vector PT7T3U 19 and Hind W -Hind m libraries of Bacillus cellulyticus K-12 chromosomal inserts created in 5.cofi. The Libraries were screened for the expression of avicelase by monitoring the immunoreaction of the anti-avicelase (immunoscreening). Positive clones (Ac-3, Ac-5, and Ac-7) contained the identical 3.5kb Hind III fragment as determined by restriction mapping and Southern hybridization, and expressed avicelase efficiently and constituvely using its own promoter in the heterologous host. From the immunoblotting analysis, a polypeptide which showed a CMCase activity with an Mr of 54000 was detected.

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Research on the Allergic Potential of Insecticidal CrylAc Proteins of Genetically Modified Rice

  • Son, Dae-Yeul
    • Food Science and Biotechnology
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    • v.15 no.3
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    • pp.385-391
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    • 2006
  • In Korea, different kinds of genetically modified (GM) crops are under development, including GM-rice expressing insecticidal crystal (Cry) proteins of Bacillus thuringiensis (Bt) modified to change a single amino acid. In this study, amino acid (aa) sequences of modified Cry proteins were compared to that of known allergens, and Cry proteins expressed in GM-rice were identified by using Cry protein specific polyclonal antibody. The antigen-antibody reactions were compared between GM and commercial rice to assess the allergic risk of Cry proteins. This analysis showed no known allergen to have more than 35% aa sequence homology with modified Cry proteins in Bt rice over an 80 aa window or to have more than 8 consecutive identical aa. Sera from allergic patients showed some IgE reactivity via immunoblotting and enzyme-linked immunosorbent assay (ELISA), although no differences were seen between GM and commercial rice. Based on these results we conclude that GM rice with modified Cry proteins has no differences in its protein composition or allergenicity relative to commercial rice.

Regulatory expression and cellular localization of doublecortin in the rat retina following ischemia-reperfusion injury

  • Gwon, Jae-Sung;Chun, Myung-Hoon;Kang, Wha-Sun
    • Animal cells and systems
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    • v.15 no.2
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    • pp.155-159
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    • 2011
  • Doublecortin (DCX) is microtubule-associated protein and is required for neuronal migration, differentiation and plasticity. In the retina, it is highly expressed between embryonic day 18 (E18) and E20, and is poorly expressed postnatally. In this study, we investigated the expression and cellular localization of DCX in the rat retina following ischemia induced by transiently increasing the intraocular pressure. While DCX immunoreactivity in control retinas was restricted to the outer border of the inner nuclear layer, it appeared in horizontal cell somata and processes in affected retinas. Quantitative evaluation by immunoblotting confirmed that DCX expression continuously increased after ischemia-reperfusion and showed 370% of control protein levels at 4 weeks after ischemic insult. These results suggest that the DCX in horizontal cells might play a role in neurite remodeling or modulating other neurons in ischemic rat retinas.

Antitumor effects of octyl gallate on hypopharyngeal carcinoma cells

  • NTK, Trang;Yoo, Hoon
    • International Journal of Oral Biology
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    • v.45 no.4
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    • pp.218-224
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    • 2020
  • The antitumor effects of octyl gallate (OG) were investigated on FaDu human hypopharyngeal squamous carcinoma cells. At various concentrations, OG inhibited the proliferation of FaDu cells by suppressing cell cycle regulators and induced apoptosis by activating caspase 3 and its downstream poly (ADP-ribose) polymerase, thereby damaging DNA. Immunoblotting demonstrated that OG significantly suppressed the expression of integrin family proteins (integrin α4, αv, β3, β4), hindering cell adhesion. The reduced expression of integrins subsequently mediated the mitogen-activated protein kinase signaling pathway to stimulate the activation of extracellular signal-regulated kinases and c-jun N-terminal kinases, leading to apoptosis. Thus, OG demonstrated antitumor activity on hypopharyngeal squamous carcinoma cells by suppressing cell proliferation and inducing apoptosis.

Esophagitis and IL-1$\beta$-induced alteration of MAP kinase activity in esophageal smooth muscle

  • Lee, Tai-Sang;Min, Young-Sil;Choi, Tae-Sik;Sim, Sang-Soo;Shin, Yong-Kyoo;Lee, Moo-Yeol;Sohn, Uy-Dong
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.117.1-117.1
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    • 2003
  • We investigated whether experimental esophagitis and IL-1$\beta$ could induce the activation of MAP kinases in esophageal smooth muscle. With two models of experimental esophagitis, we assessed the activity of p38 MAP kinase, p44/42 MAP kinase and JNK. In feline acute experimental esophagitis, immunoblotting of normal and esophagitis-induced smooth muscle with each types of MAP kinase antibodies revealed the slight increase of phosphorylated form of p38 MAP kinase, especially in membrane fraction. (omitted)

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